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https://openalex.org/W4230334467	OpenAlex	Open Science	CC-By	2,014	video 2	null	English	Spoken	871	1,847	Video 1 AP4C view showing a large ball valve thrombus in the LA. Download Video 1 via http://dx.doi.org/10.1530/ERP- 14-0064-v1 Barun Kumar MBBS MD DM, Anupam Singh MBBS MS1 and Mohamad Akram MBBS MD2 Barun Kumar MBBS MD DM, Anupam Singh MBBS MS1 and Mohamad Akram MBBS MD2 Department of Medicine (Cardiology) Himalayan Institute of Medical Sciences SRHU Dehradun Uttarakhand 1Department of Ophthalmology, AIIMS-Rishikesh, Dehradun, Uttarakhand, India 2Department of Medicine, Himalayan Institute of Medical Sciences, SRHU, Dehradun, Uttarakhand, India A 17-year-old female presented to the cardiac outpatient department with dyspnoea NYHA grade IV for the past 1 month. The patient reported a history of breathlessness for the past 5 years. On examination, the pulse rate was 90/min, regular with frequent ectopics, and BP was 90/50 mmHg. On cardiovascular examination, S1 and P2 were loud with mid diastolic murmur (MDM), pan systolic murmur (PSM) and early diastolic murmur (EDM) heard. Electrocardiography (ECG) showed normal sinus rhythm (NSM) with huge P wave. Echocardiography examination revealed severe mitral stenosis (MS) (mitral valve area (MVA): 0.65 cm2 by planimetry, Fig. 1A; a mean gradient of 33 mmHg across mitral valve, Fig. 1B), severe pulmonary artery hypertension (PAH), severe tricuspid regurgitation (TR), signiﬁcant tricuspid stenosis (TS), severe aortic regur- gitation (AR) and mild pulmonary regurgitation (PR). There was a large ball-shaped left atrium (LA) thrombus (3.65!3.55 cm), which was freely mobile and intermittently obstructing mitral oriﬁce (Fig. 2A and B; Videos 1 and 2). The patient was immediately referred for open mitral commissurotomy/mitral valve replacement (OMC/MVR), aortic valve replacement (AVR), tricuspid valve repair and LA clot removal. Figure 1 (A) Planimetry of the mitral valve and (B) pressure gradient across the mitral valve. q 2014 The authors www.echorespract.com Published by Bioscientiﬁca Ltd Downloaded from Bioscientifica.com at 10/24/2024 via Open Access. This work is licensed under a Creati Attribution-NonCommercial-NoDerivs 4.0 Internati http://creativecommons.org/licenses/b Video 2 Modiﬁed PSAX view showing a large ball valve thrombus and a clear left atrial appendage (LAA). Download Video 2 via http://dx.doi.org/10.1530/ERP-14-0064-v2 ID: 14-0064; December 2014 DOI: 10.1530/ERP-14-0064 ID: 14-0064; December 2014 DOI: 10.1530/ERP-14-0064 Large ball valve thrombus in the left atrium B Kumar and others IMAGES AND VIDEOS Open Access I10 Downloaded from Bioscientifica.com at 10/24/2024 02:07:02AM via Open Access. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. http://creativecommons.org/licenses/by-nc-nd/4.0/ Author contribution statement Dr B Kumar was the primary physician/cardiologist treating the patient. Dr A Singh helped in the preparation of the manuscript. Dr M Akram was responsible for managing the patient along with the primary cardiologist, Dr B Kumar. Declaration of interest Declaration of interest The authors declare that there is no conﬂict of interest that could be perceived as prejudicing the impartiality of the work reported. Patient consent Author contribution statement Received in ﬁnal form 23 September 2014 Accepted 9 October 2014 Funding This research did not receive any speciﬁc grant from any funding agency in the public, commercial or not-for-proﬁt sector. Figure 2 (A) Apical four chamber (AP4C) and (B) modiﬁed parasternal short axis (PSAX) view showing a large left atrium (LA) thrombus. Patient consent Written informed consent was obtained from the patient’s guardian for publication of the submitted article and the accompanying images/videos. Figure 1 Figure 1 (A) Planimetry of the mitral valve and (B) pressure gradient across the mitral valve. (A) Planimetry of the mitral valve and (B) pressure gradient across the mitral valve. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. www.echorespract.com Published by Bioscientiﬁca Ltd Downloaded from Bioscientifica.com at 10/24/2024 02:07:02AM via Open Access. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. http://creativecommons.org/licenses/by-nc-nd/4.0/ www.echorespract.com Published by Bioscientiﬁca Ltd Downloaded from Bioscientifica.com at 10/24/2024 02:07:02AM via Open Access. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. http://creativecommons.org/licenses/by-nc-nd/4.0/ q 2014 The authors Figure 2 (A) Apical four chamber (AP4C) and (B) modiﬁed parasternal short axis (PSAX) view showing a large left atrium (LA) thrombus. B Kumar and others B Kumar and others Large ball valve thrombus in the left atrium ID: 14-0064; December 2014 DOI: 10.1530/ERP-14-0064 Large ball valve thrombus in the left atrium ID: 14-0064; December 2014 DOI: 10.1530/ERP-14-0064 B Kumar and others ID: 14-0064; December 2014 DOI: 10.1530/ERP-14-0064 The patient succumbed to sudden cardiac death before surgery could be performed. This case was an unusual fatal presentation of rheumatic heart disease (RHD) with severe multivalvular involvement with a large ball valve thrombus at such a young age (1, 2, 3). References 1 Gillmer D & Campanella C 1984 Echocardiographic diagnosis of left atrial ball-valve thrombus. South African Medical Journal 65 662. 2 Raghuram AR, Kumar S & Balamurugan K 2007 Left atrial ball thrombus. Indian Journal of Thoracic and Cardiovascular Surgery 23 34–35. (doi:10.1007/s12055-007-0008-7) 3 Barwad PW, Kothari SS, Bahl VK & Yadav R 2012 Anterior mitral leaﬂet and ball valve thrombus: playing hockey in the heart. Journal of the American College of Cardiology 60 e43. (doi:10.1016/j.jacc. 2012.05.068) Figure 2 (A) Apical four chamber (AP4C) and (B) modiﬁed parasternal short axis (PSAX) view showing a large left atrium (LA) thrombus. (A) Apical four chamber (AP4C) and (B) modiﬁed parasternal short axis (PSAX) view showing a large left atrium (LA) thrombus. Received in ﬁnal form 23 September 2014 Accepted 9 October 2014 www.echorespract.com	49,012
https://openalex.org/W3000405276	OpenAlex	Open Science	CC-By	2,020	Dose reduction and withdrawal strategy for TNF-inhibitors in psoriatic arthritis and axial spondyloarthritis: design of a pragmatic open-label, randomised, non-inferiority trial	Celia A J Michielsens	English	Spoken	9,826	17,281	Dose reduction and withdrawal strategy for TNF-inhibitors in psoriatic arthritis and axial spondyloarthritis: design of a pragmatic open-label, randomised, non-inferiority trial Celia A. J. Michielsens1,2, Nadine Boers1, Nathan den Broeder1, Mark H. Wenink1, Aatke van der Maas1, Elien A. M. Mahler1, Michelle L. M. Mulder1,2, Désirée van der Heijde3, Frank H. J. van den Hoogen1,2, Lise M. Verhoef1 and Alfons A. den Broeder1,2 Abstract Background: Tumour necrosis factor inhibitors (TNFi) are effective in the treatment of patients with spondyloarthritis (SpA), including psoriatic arthritis (PsA) and axial spondyloarthritis (axSpA). However, these drugs come with some disadvantages such as adverse events, practical burden for patients and high costs. Dose optimisation of TNFi after patients have reached low disease activity (LDA) has been shown feasible and safe in rheumatoid arthritis (RA). However, data on TNFi dose optimisation in PsA and axSpA are scarce, especially pragmatic, randomised strategy studies. Methods: We developed an investigator-driven, pragmatic, open-label, randomised, controlled, non-inferiority trial (DRESS-PS) to compare the effects of a disease activity-guided treat-to-target strategy with or without a tapering attempt in patients with SpA (PsA and axSpA combined), ≥16 years of age, who are being treated with TNFi, and have had at least 6 months of low disease activity. The primary outcome is the percentage of patients in LDA after 12 months of follow up. Patients are assessed at baseline, 3, 6, 9, and 12 months of follow up. Bayesian power analyses with a weakened prior based on a similar study performed in RA resulted in a sample size of 95 patients in total. Discussion: More knowledge on disease activity-guided treatment algorithms would contribute to better treatment choices and cost savings and potentially decrease the risk of side effects. In this article we elucidate some of our design choices on TNFi dose optimisation and its clinical and methodological consequences. Trial registration: Dutch Trial Register, NL6771. Registered on 27 November 2018 (CMO NL66181.091.18, 23 October 2018). Keywords: Spondyloarthritis, Psoriatic arthritis, Dose reduction, Tapering, Randomised controlled Correspondence: c.michielsens@maartenskliniek.nl * Correspondence: c.michielsens@maartenskliniek.nl 1Department of Rheumatology, Sint Maartenskliniek, Hengstdal 3, 6574 NA Nijmegen, The Netherlands 2Department of Rheumatic Diseases, Radboud Institute of Health Sciences, Radboud University Medical Centre, Nijmegen, The Netherlands Full list of author information is available at the end of the article Michielsens et al. Trials (2020) 21:90 https://doi.org/10.1186/s13063-019-4000-5 Michielsens et al. Trials (2020) 21:90 https://doi.org/10.1186/s13063-019-4000-5 Open Access Background Tumour necrosis factor inhibiting agents (TNF-inhibi- tors, TNFi) are effective and safe in the treatment of spondyloarthritis (SpA) such as psoriatic arthritis (PsA) and axial spondyloarthritis (axSpA) [1, 2]. p y p Treatment of SpA with TNFi is often done using a more informal treat-to-target (T2T) approach, than that is used in rheumatoid arthritis (RA) [3], setting a treatment target such as low disease activity (LDA), assessing disease activ- ity and adjusting treatment if the treatment target is not reached. However, whether dose tapering or stopping once the treatment target has been reached is also part of the treatment strategy is controversial, as can be seen in the different recommendations and the American College of Rheumatology (ACR) (“we conditionally recommend against tapering of the biologic dose as a standard ap- proach”) and Assessment of SpondyloArthritis Inter- national Society-European league Against Rheumatology (ASAS-EULAR) guideline (“tapering, but not stopping a bDMARD, can be considered in patients in sustained re- mission”). These differences reflect the lack of evidence supporting these statements [4, 5]. The question to taper or not is indeed important. Des- pite its effectiveness, the use of TNFi also incorporates downsides including high costs and adverse events such as injection site reactions, an increased risk of infections and practical burden for patients [6, 7]. Therefore, it makes sense to explore strategies that optimise the risk- benefit ratio of these drugs, such as dose optimisation after patients have reached LDA. Still, prospective and randomised controlled strategy studies of dose optimisation (including stopping) in both PsA and axSpA are absent. Summarising the evidence, it seems plausible that a relevant proportion of patients with PsA or axSpA can maintain LDA following dose optimisation strategies or discontinuation of their TNFi once remission or LDA is reached, but this strategy has not been put to the test in a strategy RCT. Of note, this subject has been prioritised in the research agenda of some of the large rheumatology associations [3, 5]. In RA, T2T dose optimisation has indeed been in- cluded in the European recommendations [8]. A recent Cochrane review showed that fixed dose reduction (e.g. by 50%) and disease activity-guided dose optimisation for TNFi is comparable to continuation of treatment, al- though discontinuation without reinstatement of TNFi in the case of a flare seems to be an inferior strategy [9]. Additionally, previous studies have demonstrated the strong cost-effectiveness of tapering in RA [10–12]. Background Compared to the considerable amount of evidence on dose optimisation for TNFi in RA, data on this subject in PsA and axSpA are scarce [3, 13]. We therefore set out to perform a pragmatic, open- label, randomised, controlled, non-inferiority strategy trial comparing a TNFi T2T strategy with or without a tapering attempt. During the design of this trial, a num- ber of choices were made that we would like to address in this paper. In PsA, a systematic review summarised the limited current evidence on dose optimisation and withdrawal of biological disease-modifying anti-rheunatic drugs (bDMARDs) in PsA, and favoured dose optimisation over discontinuation because of the substantial risk of loss of remission [14]. So far, no randomised controlled trials have been performed on dose optimisation strat- egies in PsA. © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Page 2 of 12 Page 2 of 12 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (202 leads to flares [14, 15]. A few randomised controlled tri- als have been conducted on dose optimisation or discon- tinuation in axSpA, where similar results were shown as in RA with regard to maintaining remission by fixed dose reduction (e.g. by 50%) compared with full dose etanercept [16]. A randomised controlled non-inferiority trial reported by Gratacós et al. demonstrated non- inferiority between dose optimisation and full-dose TNFi treatment in patients with axSpA. However, TNFi treat- ment was reduced by 25% only, and the trial was lacking a sufficient disease activity-guided tapering algorithm [17]. A recent randomised controlled trial by Landewé et al. in patients with non-radiographic axSpA (nraxSpA) raised two important points. First, it showed that after 28 weeks of achieving inactive disease with adalimumab therapy, 47% of patients who discontinued adalimumab and switched to placebo had no flares during the follow- ing 40 weeks, supporting the possibility of (temporary) sustained remission after discontinuation of TNFi in pa- tients with nraxSpA. Furthermore, the authors state that the majority of patients who did have a flare did fully re- cover to their previous state of clinical remission after adalimumab reinstatement. The question arises whether the placebo group would have fewer flares if adalimu- mab was tapered instead of abruptly stopped. Interest- ingly, predictors for maintaining drug-free remission were not identified [18]. Methods Design The dose reduction strategy study of TNF inhibitors in psoriatic arthritis and axial spondyloarthritis patients (DRESS-PS) is a pragmatic, open-label, mono-centre, ran- domised, controlled, non-inferiority strategy trial, and is currently recruiting patients at the departments of Rheumatology of the Sint Maartenskliniek in Nijmegen and Woerden and at the department of Rheumatic Diseases of the Radboud University Medical Centre In axSpA, two systematic reviews concluded that more or less comparable to PsA, TNFi reduction strategies are successful in sustaining clinical remission or LDA in about 50% of patients but that discontinuation often Page 3 of 12 Page 3 of 12 Michielsens et al. Trials (2020) 21:90 (Radboudumc) in Nijmegen, the Netherlands (Fig. 1; Standard protocol items: recommendation for interven- tional trials (SPIRIT) checklist is shown in Add- itional file 1). The study design is inspired by the original DRESS study [19, 20]. patients with PsA and axSpA is non-inferior compared to T2T without a tapering attempt regarding the propor- tion of patients with LDA at 12 months using a pre-set non-inferiority margin of 20%. We chose having LDA at 12 months of follow up as our primary outcome. Persist- ent flare - the outcome used in the RA DRESS study - was not feasible as an outcome, since there are currently no validated flare criteria to appropriately detect persist- ent flares in PsA or axSpA, as opposed to RA (discussed Fig. 1 Standard protocol items: recommendation for interventional trials (SPIRIT) figure: trial visits and assessments. Schedule of enrolment, time point in months. Patients visit the outpatient clinic every 3 months and if necessary extra visits are warranted. TNFi, tumour necrosis factor inhibitor Objective Our definition of LDA is based on the Psoriatic Arthritis Disease Activity Score (PASDAS) and Ankylosing Spondylitis Disease Activity Score (ASDAS) criteria, but also involves the absence of extra- axial manifestations, since treatment decisions in PsA and axSpA are not solely based on the PASDAS and ASDAS but also encompass skin and eye manifestations and inflammatory bowel disease, which are not suffi- ciently covered in the PASDAS and ASDAS alone. successfully taper or discontinue TNFi use and the per- centage defined daily dose (DDD) of TNFi use; to com- pare the change in the PASDAS and ASDAS at each time point; to compare the cumulative incidence and number of flares between the T2T groups with or with- out a tapering attempt; to compare the functioning in both groups (Health Assessment Questionnaire Disabil- ity Index (HAQ-DI) and Bath Ankylosing Spondylitis Functional Index (BASFI)); to compare the proportion of patients using non-steroidal anti-inflammatory drugs (NSAIDs), corticosteroid or conventional synthetic/bio- logical/targeted synthetic disease-modifying anti- rheumatic drug (cs/b/tsDMARDs) in both groups; to com- pare the occurrence of adverse events, especially infections. In addition, we will determine the cost effectiveness of T2T with or without a tapering attempt and investigate whether baseline factors are able to predict successful dose optimisation. DMARD disease-modifying anti-rheumatic drug, bDMARD biological DMARD, TNFi tumour necrosis factor inhibitor, IL interleukin, CTLA4-Ig cytotoxic T-lymphocyte associated protein 4-immunoglobulin, PDE-4i phosphodiesterase-4 inhibitor, JAKi janus kinase inhibitor Registered dosage in psoriatic arthritis, axial spondyloarthritis and non-radiographic axial spondyloarthritis Objective The primary objective is to investigate whether a proto- colised T2T strategy of TNFi with a tapering attempt in Fig. 1 Standard protocol items: recommendation for interventional trials (SPIRIT) figure: trial visits and assessments. Schedule of enrolment, time Page 4 of 12 Page 4 of 12 Michielsens et al. Trials (2020) 21:90 later, p.17) [21]. Another option was to use the area under the curve (AUC) (mean time weighted) for disease activity. However, in our view this does not reflect the main outcome of a tapering strategy, because differences will probably occur between the two groups due to tapering-associated short-lived flares. While these short- lived flares are important, we assume that they can be treated easily, and have no sequelae like loss of function, reduced quality of life (QOL) or important radiological damage, and that being in LDA at the study end more truly reflects the main goal of trial and error tapering. Furthermore, a mean time weighted or an AUC value is difficult to interpret, and does not easily allow for com- bination of both diseases, since different disease activity measures are used. Our definition of LDA is based on the Psoriatic Arthritis Disease Activity Score (PASDAS) and Ankylosing Spondylitis Disease Activity Score (ASDAS) criteria, but also involves the absence of extra- axial manifestations, since treatment decisions in PsA and axSpA are not solely based on the PASDAS and ASDAS but also encompass skin and eye manifestations and inflammatory bowel disease, which are not suffi- ciently covered in the PASDAS and ASDAS alone. later, p.17) [21]. Another option was to use the area under the curve (AUC) (mean time weighted) for disease activity. However, in our view this does not reflect the main outcome of a tapering strategy, because differences will probably occur between the two groups due to tapering-associated short-lived flares. While these short- lived flares are important, we assume that they can be treated easily, and have no sequelae like loss of function, reduced quality of life (QOL) or important radiological damage, and that being in LDA at the study end more truly reflects the main goal of trial and error tapering. Furthermore, a mean time weighted or an AUC value is difficult to interpret, and does not easily allow for com- bination of both diseases, since different disease activity measures are used. Patients Trials (2020) 21:90 Page 5 of 12 Page 5 of 12 Page 5 of 12 different effects in the two diseases. The T2T principle is also already widely pursued in multiple chronic in- flammatory disorders, including PsA and axSpA, indicat- ing that this overarching principle seems disease agnostic [9, 17, 18, 22]. Additionally, the outcome of non-inferiority of the tapering strategy is not dependent on the percentage of patients that can taper or stop, but on the implementa- tion of the T2T strategy and the effectiveness of increased or restarted dosing on disease activity, and we do not antici- pate effect modification between the two closely related diseases. deemed exclusion of patients with inactive extra-axial manifestations unnecessary since in the psoriasis tapering trial patients seem to respond well to tapering strategies [23] and a similar study in patients suffering from IBD is currently being conducted [24]. We exclude only patients whose comorbidity could inter- fere with our protocolised dose optimisation strategy, thus being unable to participate due to the required treatment with TNFi (e.g. active Crohn’s disease, ulcerative colitis, uveitis, psoriasis), or when it is expected that the outcome cannot be measured (short life expectancy, planned major surgery). Pregnant women are also excluded from participa- tion in this trial. A previous (successful or unsuccessful) at- tempt at dose optimisation is allowed if attempted more than 2 years ago. The use of concomitant NSAIDs and csDMARDs is allowed before and during participation in this study, though the intake of NSAIDs has to have been stable at least 8 weeks before inclusion. Patients with PsA or axSpA diagnosed clinically by the treating rheumatologist (and supported by Classification Criteria for Psoriatic Arthritis (CASPAR) and ASAS classification criteria) are included. To have optimal generalizability of our study to daily clinical practice, we pragmatically decided to keep the amount of inclusion and exclusion criteria as limited as possible. These pa- tients are eligible if they have LDA up to 6 months prior to inclusion and are using > 50% of the authorized DDD of a TNFi (Table 2). Treatment decisions are made based on objective and subjective disease activity scores, by shared decision making between rheumatologist and patients. Patients In patients with PsA, stable LDA is defined as having a Psoriatic Arthritis Disease Activity Score (PAS- DAS) ≤3.2 and body surface area involvement (modified BSA) ≤3%, used as a target by rheumatologists in routine practice and in the minimal disease activity (MDA). In axSpA this is defined as an Ankylosing Spondylitis Disease Activity Score (ASDAS) < 2.1 in PsA and axSpA an ab- sence of active extra-axial disease-related symptoms caused by Crohn’s disease, ulcerative colitis, uveitis or psoriasis is also required for LDA, or when formal mea- surements are not available, the judgement of the phys- ician and patient can be used instead. Patients with extra- axial manifestations of disease, such as inflammatory bowel disease (IBD), uveitis or psoriasis are included in our study, unless currently active extra-axial manifesta- tions prevent dose optimisation. If extra-axial symptoms develop during treatment, physicians are allowed to treat them by delaying tapering or increasing TNFi dosage. We We decided to include not only patients in remission, but extended this to patients with LDA 6 months prior to participation, for two reasons. First, this increases both the generalizability of the study and the feasibility, since many physicians and patients do not strive for re- mission, and there are no data on the diseases showing that a treatment target of remission results in better out- comes than a treatment target of LDA. Second, in stud- ies with RA, there is no evidence that baseline disease activity is strongly associated with a chance of successful tapering [25], so this choice is not expected to jeopardize internal validity due to introducing effect modification. TNFi tumour necrosis factor inhibitor Patients We have chosen to combine PsA and axSpA in this study mainly for feasibility reasons. Furthermore, PsA and axSpA share pathophysiological, genetic and clinical characteristics, and as current treatment options are al- most identical with respect to type of drugs used, dosing and concomitant DMARDs used (Table 1) and finally because preliminary dose optimisation data are similar, we felt this was possible without too much risk of Several secondary outcomes will be compared between the two groups, including disease activity, flares, func- tion, quality of life, costs, co-medication and safety. Our main secondary objectives are to assess the proportion of patients in the intervention group that can Table 1 Overview of DMARDs in psoriatic arthritis, radiographic axial spondyloarthritis and non-radiographic axial spondyloarthritis Psoriatic arthritis Radiographic axial spondyloartritis Non-radiographic axial spondyloarthritis bDMARDs TNFi Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Infliximab 5 mg/kg 1x/8 weeks Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Infliximab 5 mg/kg 1x/8 weeks Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Anti-IL-17 Secukinumab 150 mg 1x/month (up to 300 mg) Ixekizumab 80 mg 1x/4 weeks Secukinumab 150 mg 1x/month Anti-IL-23, IL-12 Ustekinumab 45 mg 1x/12 weeks CTLA4-Ig Abatacept 125 mg 1x/week PDE-4i Apremilast 30 mg 2x/day JAKi Tofacitinib 5 mg 2x/day DMARD disease-modifying anti-rheumatic drug, bDMARD biological DMARD, TNFi tumour necrosis factor inhibitor, IL interleukin, CTLA4-Ig cytotoxic T-lymphocyte associated protein 4-immunoglobulin, PDE-4i phosphodiesterase-4 inhibitor, JAKi janus kinase inhibitor Registered dosage in psoriatic arthritis, axial spondyloarthritis and non-radiographic axial spondyloarthritis Table 1 Overview of DMARDs in psoriatic arthritis, radiographic axial spondyloarthritis and non-radiographic axial spondyloarthritis Psoriatic arthritis Radiographic axial spondyloartritis Non-radiographic axial spondyloarthritis Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Infliximab 5 mg/kg 1x/8 weeks Secukinumab 150 mg 1x/month Adalimumab 40 mg 1x/2 weeks Certolizumab 200 mg 1x/2 weeks Etanercept 25 mg 2x/week or 50 mg 1x/week Golimumab 50 mg 1x/month Infliximab 5 mg/kg 1x/8 weeks Secukinumab *150 mg 1x/month Michielsens et al. Intervention Participants are randomised using a computer-generated procedure, stratified by disease and for use of csDMARDs, to ensure equal distribution and prevent bias. We did not stratify by the different types of TNFi, since we expect a similar mechanism of action while ta- pering as demonstrated in previous studies [9]. Studies on dose optimisation with different types of TNFi, though mainly including data on adalimumab and etanercept, show no major difference in clinical outcome after dose optimisation or stopping. Patients are randomised in a ra- tio of 2:1 to the dose T2T strategy with or without a taper- ing attempt, respectively. The intervention group is larger to ensure that a more potential predictive factors for response can be studied in multivariate prediction model- ling in the dose optimisation group. Randomisation blocks using variable block sizes (multiples of 3 or 6) are used to more closely achieve the intended allocation ratio and to prevent the allocation being predictable for the treating rheumatologist. Patients, physicians, nurses and re- searchers are not blinded during this study. Analyses are blinded to treatment allocation. The treatment in the intervention group is identical to the control group with the addition of a tapering attempt, leading to a trial and error dose-optimisation and eventu- ally discontinuation. The interval of TNFi administration is extended gradually every 3 months in patients in the intervention group. Patients using the full dose discon- tinue the TNFi at approximately 6 months and are closely monitored for another 6 months, for a study total duration of 12 months. The dose optimisation steps from 100% to 66% and 50% before stopping for the different TNFi are depicted in Table 2. If a patient is using another dose regi- men than the one proposed above, an alternative dose op- timisation strategy is used. Patients who are not receiving full dosage of their TNFi step in at the nearest dosing interval. Patients encountering a flare are treated accord- ingly, following the same standardised protocol as the control group. All patients experiencing a flare are seen 4 weeks later. Should a flare persist, after receiving other medication, the dose is adjusted to the last effective inter- val or dosage. During this month patients do not continue dose optimisation but maintain the last effective interval or dosage. Interventions C t l Patients who are allocated to the control group continue their treatment by their treating rheumatologist based on a standardised treatment protocol. Patients visiting the outpatient clinic every three months are treated ac- cording to the T2T principle without a tapering attempt. Disease activity is measured at every visit, and we aim to achieve PASDAS ≤3.2 in patients with PsA and ASDAS < 2.1 in patients with axSpA. Extra visits are warranted if pa- tients report complaints related to an increase in disease activity in between regular visits. The treating rheumatolo- gist receives advice from the research physician at every Patient recruitment All eligible patients are selected and approached based on information from the electronic health record according to the aforementioned inclusion criteria. Pa- tients are asked by their treating rheumatologist to join this study, by letter accompanied by the patient informa- tion sheet and the informed consent letter. Patients re- ceive this information two weeks prior to a planned outpatient clinic visit. At the outpatient clinic visit (which is referred to as visit −1), the study is discussed and informed consent is obtained. Ethical considerations The study received ethical approval from the CMO region Arnhem Nijmegen (NL66181.091.18) and has been regis- tered in the Dutch Trial Register (NTR 7640). Privacy of patients is protected according to Dutch law AVG (Alge- mene Verordening Gegevensbescherming), by using anon- ymized data and restricting access to patient identification logs. We established a Data Safety Monitoring Board (DSMB) where four independent DSMB members, of whom two are rheumatologists, one a pharmacist and one Table 2 Dose optimisation strategy for TNFi TNFi 100% 66% 50% 0% Adalimumab/Certolizumab 40 mg 14 days interval 40 mg 21 days interval 40 mg 28 days interval Stop TNFi Etanercept 50 mg 7 days interval 50 mg 10 days interval 50 mg 14 days interval Stop TNFi Golimumab 50 mg 4 weeks interval 50 mg 6 weeks interval 50 mg 8 weeks interval Stop TNFi Infliximab 5 mg/kg 8 weeks interval 3 mg/kg 8 weeks interval 3 mg/kg 8 weeks interval 2.25 mg/kg 8 weeks interval 1.5 mg/kg 8 weeks interval 1.5 mg/kg 8 weeks interval Stop TNFi TNFi tumour necrosis factor inhibitor Michielsens et al. Trials (2020) 21:90 Page 6 of 12 Page 6 of 12 visit, on what should be done according to the protocol. However, all treatment choices are left to the discretion of the treating rheumatologist in shared decision making with the patient. When patients experience a flare, in spite of the maximal TNFi dose, treatment is adapted accord- ingly by the rheumatologist to ensure optimal care. This might include adding NSAIDs, especially in axSpA, or use of glucocorticoids. Patients visit the outpatient clinic after 4 weeks to evaluate the effects of the treatment administered. In the case of a persistent flare, patients are switched to an- other TNFi or non-TNFi class of bDMARDs as per our treatment protocol. Since treatment changes are based on shared decision making between patient and physician in daily clinical practice, in patients experiencing flare and not meeting the proposed flare criteria, treatment can be inten- sified nevertheless. Dose optimisation or discontinuation for reasons other than adverse events is discouraged but allowed. a biomedical scientist, will discuss and review data on re- cruitment, efficacy, safety, protocol adherence and proto- col updates on Good Clinical Practice (GCP). The trial is investigator-driven, and funded by ReumaNederland (funding number 17–3-303). Intervention When the latter does not suffice to improve the disease activity, the patient is advised to switch to another bDMARD following the standardised treatment protocol. Outcomes As main disease activity measures in the study, we have chosen the PASDAS (for PsA) and ASDAS (for axSpA). Since a subset of patients with PsA and AxSpA have extra-axial manifestations of disease, we also assess the occurrence of these manifestations and patient and physician opinion on disease activity. Several known disease-specific measures for PsA were reviewed and screened for reliability, validity and Page 7 of 12 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 feasibility. In the process of selecting a suitable disease activity measurement tool for PsA in this trial, certain criteria had to be met in order to be feasible for use in routine clinical practice. Furthermore, the disease activ- ity measure had to cover different domains of the disease and should be able to distinguish between different levels of disease activity, using clear cut-off points for disease activity and improvement. PASDAS > 3.2 and ASDAS ≥2.1, has an increase in dis- ease activity (≥0.8 points of PASDAS for PsA or 0.9 for axSpA), has an important worsening of mBSA, or in the occurrence of active extra-axial symptoms as judged by the treating rheumatologist, with a duration longer than 1 month. Since treatment changes are based on shared decision making between patient and physician in daily clinical practice, if patients experience flare and do not meet the proposed flare criteria, treatment can be inten- sified nevertheless. y p PsA activity often includes pain or swelling of the ankles and distal interphalangeal joints in the hands, which makes Disease Activity Score 28-C-reactive protein (DAS28-CRP) a less suitable measurement tool, since it only measures 28 joints and does not include evaluation of these joints. The Disease Activity in PSoratic Arthritis (DAPSA) does not have a normal distribution and is not sensitive enough to change, having less favourable clinimetric properties com- pared to PASDAS [26]. A third option is the use of the minimal disease activity (MDA) criteria, as LDA as calcu- lated by PASDAS is comparable with MDA. However, the MDA criteria result in a binominal score (yes/no), which provides less information than for example the PASDAS. Furthermore, since the MDA includes scoring of dermato- logical manifestations, it is less practical to use in daily clin- ical practice. Assay sensitivity Si hi i In axSpA, the BASDAI and ASDAS are both acceptable disease activity measurement tools. We decided to use the ASDAS, since it also includes an objective measure with CRP [5] and has an established longitudinal relationship with structural damage, supporting its construct validity. Also, several cut-off values have been well-validated, in- cluding minimal clinically important worsening (MCIW). Since this is a non-inferiority trial, assay sensitivity, e.g. the ability of the trial to detect inferiority if it is present, is important to take into account. Assay sensitivity can be supported by a third arm in which a clearly inferior treatment is tested leading indeed to the conclusion of inferiority. In our study this would for example consti- tute an arm in which TNFi are not reinstated when flares occur. However, for several reasons we chose not to include a third arm. First, we use validated outcome measures (PASDAS and ASDAS) with known sensitivity to change, and several superiority studies have shown that these can detect differences between placebo and active treatment. Second, similar trials in RA and psoria- sis have indeed proven inferiority of dose optimisation As mentioned before, patients are frequently asked about the occurrence of active extra-axial disease manifestations such as Crohn’s disease, ulcerative colitis and uveitis. Outcomes Last, the Composite Psoriatic Disease Activity Index (CPDAI) seems to have less discriminative ability in distinguishing high and low disease activity than the other outcome measures [26–29]. Therefore, we opted to use PASDAS as a disease-specific activity measure for PsA, due to the following reasons: it is a continuous disease activity index that includes arthritis (66/68 joint score), dactylitis, enthesitis, physician disease activity visual analogue scale (VAS) score, patient disease activity VAS, patient-reported physical function and CRP [28]. To cover the dermato- logical component of PsA in our trial, psoriasis is measured by modified BSA (mBSA), since an increase in psoriasis ac- tivity is not sufficiently covered by the PASDAS. The mBSA ranges from 0 to 1 and 2 and correlates with no skin mani- festations to ≤3% and > 3% of the body surface area covered. PsA flare is defined as an increase in PASDAS ≥0.8 points, since this increase equals the measurement error of the instrument [30]. Choosing 1–2 times the measure- ment error to define the MCIW is comparable to other in- struments such as the DAS28-CRP used in RA and ASDAS in axSpA. In the dermatological domain, clear cut-off values for important worsening are lacking for mBSA and treatment is adjusted as judged by the treating rheumatologist and patient, since this is a very individual choice. AxSpA flare is defined as an increase in ASDAS ≥ 0.9 (the published MCIW for ASDAS) [31, 32]. We con- sider our flare criteria rather sensitive and therefore think we can detect any significant flare. Non-inferiority margin The choice for a suitable non-inferiority (NI) margin is essential for a NI trial to provide clinically relevant con- clusions [33, 34]. We chose a NI margin of 20%. In the DRESS study, 16% of patients were successfully stopped and 45% tapered, meaning 60% benefitted from a dose optimisation strategy, resulting in a number needed to treat (NNT) of about 1.5. If the confidence interval (CI) around the point estimate of the proportion of patients in LDA indeed remains below 20%, this strikes an ac- ceptable balance between the advantages of dose opti- misation such as reduced injection burden and risk of side effects, and the harm incurred from loss of LDA (with a substantially higher NNH of 5). Data analyses l Primary analyses will be per-protocol (PP), as this is the most conservative for a non-inferiority study. Addition- ally, analyses are performed on an intention-to-treat basis (ITT). For PP analysis, we include intervention pa- tients that attempted at least one dose optimisation step and control patients who did not attempt dose optimisa- tion for reason of treatment relaxation per se, but only when medically required such as in the case of adverse events or contraindications. Descriptive statistics are provided using mean and standard deviation (SD), me- dian (p25-p75) or frequencies/percentages depending on the type of distribution of the data. For exclusion and dropout, numbers and reasons are reported to ensure in- ternal validity. Missing values are imputed using mul- tiple imputations when meeting the assumption of missing completely at random (MCAR)/missing at ran- dom (MAR), as imputation always increases precision and often also reduces bias [37, 38]. As motivated above, we take the non-inferiority mar- gin of −0.2 (20%) and the randomisation ratio of 2:1 of intervention versus control. The ratio of 2:1 for interven- tion and control sample size is chosen to be able to include more determinants in a prediction model for successful dose optimisation. The sample size of the study is based on a Bayesian analysis where non- inferiority will be claimed if the lower limit of the Bayesian 95% credibility interval of the difference is above −20%. We chose a Bayesian sample size approach to be able to include information from the original DRESS study, resulting in a prior tending towards non- inferiority of dose optimisation [20]. This will enable us to take into account the previous evidence on TNFi ta- pering and reduces the required sample size without compromising the power of our study. Definition of flare Since validated criteria for persistent flare are lacking, we consider patients to have a flare in several situations: when a patient experiences loss of LDA as defined by Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 Page 8 of 12 To make sure this prior based on the previous study does not have too much weight in the Bayesian analysis, since RA is a different disease, the precision of this prior is reduced and the prior is taken as normally distributed with mean −2% and standard deviation of 22.5%. Each percentage is estimated from 500 simulated datasets of the control group having an LDA proportion of 80%. Bayesian analysis is performed with a generalized linear model with identity link and binomial distribution using a normally distributed prior with mean −2% and stand- ard deviation of 22.5%. We will recruit 90 subjects in total (30 vs 60) so as to have 80% power to claim infer- iority when the true difference is 0%. Adding 5% drop- out, the total number of patients needed is 95 patients. Although this is the minimum number of patients needed, we will attempt to include this number of pa- tients for both PsA and axSpA separately, to achieve suf- ficient power for an analysis stratified by disease. with regard to several outcomes, both primary and sec- ondary. In the DRESS study inferiority was shown on short flares, minimal radiographic progression and AUC DAS28 [20], while STRASS was inferior on the DAS28 score at 18 months [35]. In psoriasis, a different disease entity, the CONDOR study showed inferiority on the PASI score [23]. Considering this, we feel that assay sen- sitivity of our trial design is sufficiently supported, and find it both unethical and impractical to randomise pa- tients into an arm in which medication is not reinstated, as it would induce more patient harm, and require a substantially larger sample size. Sample size considerations and statistical analyses We assume that both treatment arms have a prevalence of LDA state of 0.8 (80%) at 12 months, thus assuming 20% will lose LDA in both groups, as expected due to re- gression to the mean, based on previous research and clinical practice [18, 20]. We expect that dose optimisa- tion will cause more (short-lived) flares, however we do not anticipate a permanent loss of LDA state after a T2T strategy with compared without a tapering attempt, because of the short longevity of the flares before re- instatement of the TNFi. Discussion Considering their similarities, for ex- ample, it could be argued that PsA and psoriasis treatment trials could be performed combining the dis- eases, but since these diseases are treated by different specialists and the treatment often showcases a different effect on skin and joints, this is not usually done. In this case, considering the feasibility, similarities in treatment, the existence of validated disease measurement scores, the possibility of tapering in a similar relevant propor- tion of patients and recovering to a previous state of clinical remission after reinstatement, patients are ana- lysed in one group. witnessed by the recent 80% + discounts on adalimumab offered by AbbVie in Europe and the Netherlands espe- cially [44, 45]. As more outpatient visits are required during tapering and more time has to be invested in treating flares, it remains to be seen how the cost effect- iveness of dose optimisation strategies evolves over time. However, costs associated with side effects like infections might also play a role, which could be a burden and cause work productivity loss. Also, besides cost reduc- tion, benefits of dose optimisation include reduced pa- tient burden due to self-injection and lower risks of adverse events. These benefits alone warrant the explor- ation of dose optimising strategies. p g g Moreover, given the benefits, the question arises as to who is responsible for the execution of dose optimisa- tion research. Pharmaceutical companies often perform efficacy trials and are usually obliged to monitor infor- mation about the drugs’ safety and efficacy in so-called phase IV studies after authorization. However, when implementation of a certain treatment is established, re- assessment of the efficacy in the case of dose optimisa- tion or discontinuation is often neglected. Therefore, many dose optimisation studies are currently financed by public funds. When such trials are executed by pharmaceutical companies, the study designs tend to favour continuation of treatment, with the results fol- lowing suit. For example, the choice not to follow a T2T treatment strategy (or at least reinstate therapy in the case of flare) and lack of cost-effectiveness analyses. Fur- thermore, it is expected that patients benefit from as little exposure to medication as possible following a dose-activity-guided strategy. Therefore, it could be de- bated whether it is the pharmaceutical company’s responsibility to conduct well-designed phase III/IV dose-reduction studies for authorized drugs, e.g. after several years of use. Discussion Recent developments in this field in- clude that pharmaceutical companies receiving approval for the use of TNFi in nraxSpA are obliged by the Euro- pean Medicines Agency (EMA) to perform tapering studies, as witnessed by, for example, the data from the ABILITY 3 study. y g p Another interesting point that can be disputed is whether these kind of T2T studies produce meaningful results. After all, diseases that can be measured, and treated with effective interventions should always be eli- gible for T2T strategies that ultimately will lead to re- sults that are identical to any alternative targeted strategy. Flares or relapses of disease can after all be countered by (temporarily) increasing treatment dosages. This yields similar disease activity scores in the com- pared groups in the long run. Indeed, this is seen in many RA T2T studies in which all treatment arms con- verge after a certain amount of time to similar outcomes [39–43]. Proving non inferiority may therefore be a re- dundant goal, as per definition, this will always be achieved, yielding identical results in both groups. More evidently, if such a T2T study fails to prove non- inferiority, this must mean the execution of T2T was suboptimal, and this of course leads to circularity by def- inition. Of note, the prerequisite is that the T2T strategy study is adequately performed and sufficiently powered. The STRASS study, for example, failed to show non- inferiority in the T2T strategy due to insufficient recruit- ment [35]. However, the proof of the pudding is in the eating, and although logically sound, a strategy using T2T should be tested in a controlled trial, if only to demonstrate the feasibility of achieving such a level of treat-to-target. A final consideration is the open-label strategy. A gen- eral disadvantage of open-label studies is the risk of ex- pectation bias (placebo/nocebo effect). However, since this trial studies a disease activity-guided strategy, which can result in flares rather than response to treatment, expectation bias favouring tapering is not likely. Since this is a non-inferiority design, expectation bias would rather result in inferiority of the tapering group, result- ing in a more conservative estimation of the effect. Open-label tapering studies in RA indeed show that suc- cessful dose optimisation is achieved in a smaller pro- portion of patients compared to similar blinded studies [9]. Discussion As we are only interested in claiming non-inferiority of a T2T strategy with versus without a tapering at- tempt, we only want to control the probability of claim- ing non-inferiority when inferiority is present. Taking into account the prior evidence, favouring non- inferiority of dose reduction, we choose to control this probability at 10% instead of 5% in line with the Food and Drug Administration (FDA) guidance on Bayesian statistics, which recommends a less stringent control of frequentist type I error if prior information is favourable [36]. In the RA DRESS study, a difference in persistent flare (which would lead to loss of LDA at 12 months) of −2% was observed (90% LDA in 59 patients on continu- ation and 88% in 121 patients with dose optimisation). In addition to the motivations we provide for all the de- sign choices we made in our study, we want to discuss some important overarching questions. We have decided to include patients with PsA and axSpA combined in one trial, as we argue that these disease entities are suffi- ciently similar and treated with similar TNFi to be ana- lyzed in one composite group, thus also making the study more feasible. This leads to an interesting ques- tion: when are groups of patients similar or different enough to warrant separate studies? There are many ex- amples that ask for a similar judgement call. In RA, for example, rheumatoid factor positive (RF+) and rheuma- toid factor negative (RF-) patients are both included in the majority of studies, while pathophysiologically, they Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 Page 9 of 12 Page 9 of 12 might be very different. Other examples of diseases stud- ied that may have less in common than PsA and axSpA are abundant; granulomatosis with polyangiitis (GPA), with myeloperoxidase- (MPO) or proteinase 3- (PR 3), or anti-neutrophil cystoplasmic antibody (ANCA), pneu- moniae with several causative agents, or studies on IBD including both Crohn’s disease and ulcerative colitis. De- ciding to combine or split patients with different charac- teristics within a family of diseases remains a matter of specific judgement, especially whether considerable effect modification is to be expected on the primary out- come measure. Competing interests DvdH has received the following consulting fees: AbbVie, Amgen, Astellas, AstraZeneca, BMS, Boehringer Ingelheim, Celgene, Cyxone, Daiichi, Eisai, Eli- Lilly, Galapagos, Gilead, Glaxo-Smith-Kline, Janssen, Merck, Novartis, Pfizer, Regeneron, Roche, Sanofi, Takeda, UCB Pharma, Director of Imaging Rheuma- tology bv. AdB has received congress invitations from Roche, Cellgene, Pfizer, Abbvie, Biogen, and expert witness fees from Boehringer, BMS, Amgen, Fese- niu, Novartis. The other authors declare that they have no competing interests. Abbreviations ACR A i C Abbreviations ACR: American College of Rheumatology; ANCA: Anti-neutrophil cystoplasmic antibody; AS: Ankylosing spondylitis; ASAS: Assessment of SpondyloArthritis International Society; ASAS-HI: Assessment of SpondyloArthritis International Society Health Index; ASDAS: Ankylosing Spondylitis Disease Activity Score; AUC: Area under the curve; AVG: Algemene Verordening Gegevensbescherming; axSpA: Axial spondyloarthritis; BASFI: Bath Ankylosing Spondylitis Functional Index; bDMARD: Biological disease-modifying anti-rheumatic drug; BSA: Body surface area; CASPAR: Classification Criteria for Psoriatic Arthritis; CMO: Commissie Mensgebonden Onderzoek; CRP: C-reactive protein; csDMARD: Conventional synthetic disease modifying anti-rheumatic drug; CTLA4-Ig: Cytotoxic T-Lymphocyte Associated Protein 4-Immunoglobulin; DAPSA: Disease activity in psoriatic arthritis; DAS28-CRP: Disease Activity Score 28-C-reactive protein; DDD: Defined daily dose; DMARD: Disease- modifying anti-rheumatic drug; DRESS: Dose reduction strategy study; DRESS-PS: Dose reduction strategy study of TNF inhibitors in psoriatic arthritis and axial spondyloarthritis; DSMB: Data Safety Monitoring Board; EMA: European Medicines Agency; EULAR: European League Against Rheumatism; EuroQoL-5D-3 L: Health status; GCP: Good Clinical Practice; GDPR: General Data Protection Regulation; GPA: Granulomatosis with polyangiitis; HAQ-DI: Health Assessment Questionnaire Disability Index; IBD: Inflammatory bowel disease; ITT: Intention to treat; JAKi: Janus Kinase inhibitor; LDA: Low disease activity; MAR: Missing at random; mBSA: Modified body surface area; MCAR: Missing completely at random; MCIW: Minimal clinically important worsening; MDA: Minimal disease activity; mNY: Modified New York criteria; MPO: Myeloperoxidase; MSASSS: Modified Stoke Ankylosing Spondylitis Spine Score; NI: Non-inferiority; NNH: Number needed to harm; NNT: Number needed to treat; NraxSpA: Non-radiographic axial spondyloarthritis; NSAID: Non-steroidal anti-inflammatory drug; NTR: Nederlands Trial Register; OMERACT: Outcome measures in rheumatology; PASDAS: Psoriatic Arthritis Disease Activity Score; PDE- 4i: Phosphodiesterase-4 inhibitor; PP: Per protocol; PR3: Proteinase 3; PsA: Psoriatic arthritis; QOL: Quality of life; RA: Rheumatoid arthritis; Radboudumc: Radboud University Medical Centre; RCT: Randomised Controlled Trial; RF-: Rheumatoid factor negative; RF+: Rheumatoid factor positive; SD: Standard deviation; SF-12: Short Form-12 Health Survey; SI- joint: Sacroiliac joint; SpA: Spondyloarthritis; STRASS: Spacing of TNF-blocker injections in rheumatoid arthritis study; TNFi: Tumor necrosis factor inhibitors; Supplementary information l f Supplementary information accompanies this paper at https://doi.org/10. 1186/s13063-019-4000-5. Availability of data and materials The data collected during the study are available to researchers who provide a methodologically sound proposal to achieve aims in the approved proposal or for individual participant data meta-analysis. To gain access, data requestors will need to sign a data access agreement. Authors’ contributions CM is MD and the PhD student involved in completion of the protocol, drafting the manuscript and execution of the study, and performed CMO admission and trial registration. NB is a BSc and designed and drafted the study protocol. NdB is a MSc and PhD student and designed and drafted the study protocol and contributed to statistical considerations. MW is rheumatologist and pathobiologist, specialized in translation research. AvdM is rheumatologist and involved in protocol design. EM is rheumatologist and epidemiologist involved in protocol design. MM is MSc and responsible for implementing the PASDAS in daily clinical practice. DvdH is rheumatologist and professor of the rheumatology centre of Leiden University Medical Center. FvdH is a rheumatologist and professor of the rheumatology centre of Sint Maartenskliniek and Radboudumc and involved as study sponsor, in trial rationale, drafting, patient inclusion and organisation. LV is MSc and PhD candidate involved in drafting the manuscript. AdB is rheumatologist and clinical epidemiologist, principal investigator and initiator of this RCT, and was involved in all aspects of the study and writing of the manuscript. All authors read and approved the final manuscript. Funding The study is funded by ReumaNederland, a health fund covering different topics on rheumatism. There are no other financial or non-financial conflicts of interest other than specified. The Department of Rheumatology at Sint Maartenskliniek fulfills the role as sponsor and is the coordinating study centre. Furthermore, an independent data safety and monitoring board is installed. Additional file 1. SPIRIT 2013 Checklist: Recommended items to address in a clinical trial protocol and related documents. Trial status The trial started with recruitment on 9 January 2018 and will be completed by approximately 29 January 2021. Protocol version 1.4, 01-09-2019. Discussion This may in part be due to the patient’s fear of One of the main goals of tapering strategies is to re- duce healthcare costs. However, costs of TNFi have already lowered - at least in Europe - with the introduc- tion of biosimilars, which results in price competition, as Page 10 of 12 Page 10 of 12 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 tsDMARD: Targeted synthetic disease-modifying anti-rheumatic drug; VAS: Visual analogue scale tsDMARD: Targeted synthetic disease-modifying anti-rheumatic drug; VAS: Visual analogue scale decreasing their dose and experiencing a flare, leading to the nocebo effect and incorrect attribution of complaints to the dose optimisation. However, since in clinical prac- tice tapering is not blinded either, open-label studies may more realistically reflect chances of success. Success of tapering depends not only on pharmacological factors, but as much on psychological factors, such as thorough information from the physician, or inter-patient variabil- ity in fear of flares. Therefore, open-label tapering results in an approximation of the effect in daily clinical prac- tice but possibly an underestimation of the full pharma- cological possibility. Ethics approval and consent to participate The study has received ethical review board approval (number NL66181.091.18), dated 26 June 2018. Ethics approval was obtained for all participating centres from the central Commissie Mensgebonden Onderzoek (committee of human research) CMO regio Arnhem Nijmegen, Radboud University Medical Center, PO Box 9101, 6500 HB, Nijmegen, The Netherlands. Patient informed consent will be obtained from all participants in the study, prior to inclusion. The trial is registered at Dutch Trial Register, NL6771, date 27 November 2018. References 1. Ward MM, Deodhar A, Akl EA, Lui A, Ermann J, Gensler LS, et al. American College of Rheumatology/Spondylitis Association of America/ Spondyloarthritis Research and Treatment Network 2015 Recommendations for the treatment of ankylosing spondylitis and nonradiographic axial spondyloarthritis. Arthritis Rheumatol. 2016;68(2):282–98. 19. den Broeder AA, van Herwaarden N, van der Maas A, van den Hoogen FH, Bijlsma JW, van Vollenhoven RF, et al. Dose reduction strategy of subcutaneous TNF inhibitors in rheumatoid arthritis: design of a pragmatic randomised non inferiority trial, the DRESS study. BMC Musculoskelet Disord. 2013;14:299. 2. Gossec L, Smolen JS, Ramiro S, de Wit M, Cutolo M, Dougados M, et al. European League Against Rheumatism (EULAR) recommendations for the management of psoriatic arthritis with pharmacological therapies: 2015 update. Ann Rheum Dis. 2016;75(3):499–510. 20. van Herwaarden N, van der Maas A, Minten MJ, van den Hoogen FH, Kievit W, van Vollenhoven RF, et al. Disease activity guided dose reduction and withdrawal of adalimumab or etanercept compared with usual care in rheumatoid arthritis: open label, randomised controlled, non-inferiority trial. BMJ. 2015;350:h1389. 3. Smolen JS, Schols M, Braun J, Dougados M, FitzGerald O, Gladman DD, et al. Treating axial spondyloarthritis and peripheral spondyloarthritis, especially psoriatic arthritis, to target: 2017 update of recommendations by an international task force. Ann Rheum Dis. 2018;77(1):3–17. 3. Smolen JS, Schols M, Braun J, Dougados M, FitzGerald O, Gladman DD, et al. Treating axial spondyloarthritis and peripheral spondyloarthritis, especially psoriatic arthritis, to target: 2017 update of recommendations by an international task force. Ann Rheum Dis. 2018;77(1):3–17. 4. Ward MM, Deodhar A, Gensler LS, Dubreuil M, Yu D, Khan MA, et al. 2019 Update of the American College of Rheumatology/Spondylitis Association of America/Spondyloarthritis Research and Treatment Network Recommendations for the Treatment of Ankylosing Spondylitis and Nonradiographic Axial Spondyloarthritis. Arthritis Rheumatol. 2019;71(10): 1599-613. 21. van der Maas A, Lie E, Christensen R, Choy E, de Man YA, van Riel P, et al. Construct and criterion validity of several proposed DAS28-based rheumatoid arthritis flare criteria: an OMERACT cohort validation study. Ann Rheum Dis. 2013;72(11):1800–5. 22. Coates LC, Helliwell PS. Treating to target in psoriatic arthritis: how to implement in clinical practice. Ann Rheum Dis. 2016;75(4):640–3. 5. van der Heijde D, Ramiro S, Landewe R, Baraliakos X, Van den Bosch F, Sepriano A, et al. 2016 update of the ASAS-EULAR management recommendations for axial spondyloarthritis. Ann Rheum Dis. 2017;76(6): 978–91. 23. References Atalay S, van den Reek J, van Vugt LJ, Otero ME, van de Kerkhof PCM, den Broeder AA, et al. Tight controlled dose reduction of biologics in psoriasis patients with low disease activity: a randomized pragmatic non-inferiority trial. BMC Dermatol. 2017;17(1):6. 6. Minozzi S, Bonovas S, Lytras T, Pecoraro V, Gonzalez-Lorenzo M, Bastiampillai AJ, et al. Risk of infections using anti-TNF agents in rheumatoid arthritis, psoriatic arthritis, and ankylosing spondylitis: a systematic review and meta- analysis. Expert Opin Drug Saf. 2016;15(sup1):11–34. 24. Clinical Trials: The European Union Clinical Trials Register. https://www. clinicaltrialsregister.eu/ctr-search/trial/2016-003321-42/NL. Accessed 20 August 2019. 25. Tweehuysen L, van den Ende CH, Beeren FM, Been EM, van den Hoogen FH, den Broeder AA. Little evidence for usefulness of biomarkers for predicting successful dose reduction or discontinuation of a biologic agent in rheumatoid arthritis: a systematic review. Arthritis Rheumatol. 2017;69(2): 301–8. 7. Singh JA, Cameron C, Noorbaloochi S, Cullis T, Tucker M, Christensen R, et al. Risk of serious infection in biological treatment of patients with rheumatoid arthritis: a systematic review and meta-analysis. Lancet. 2015; 386(9990):258–65. 8. Smolen JS, Landewe R, Bijlsma J, Burmester G, Chatzidionysiou K, Dougados M, et al. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2016 update. Ann Rheum Dis. 2017;76(6):960–77. 26. Salaffi F, Ciapetti A, Carotti M, Gasparini S, Gutierrez M. Disease activity in psoriatic arthritis: comparison of the discriminative capacity and construct validity of six composite indices in a real world. Biomed Res Int. 2014;2014: 528105. 9. Verhoef LM, van den Bemt BJ, van der Maas A, Vriezekolk JE, Hulscher ME, van den Hoogen FH, et al. Down-titration and discontinuation strategies of tumour necrosis factor-blocking agents for rheumatoid arthritis in patients with low disease activity. Cochrane Database Syst Rev. 2019;5:CD010455. 27. Perruccio AV, Got M, Li S, Ye Y, Gladman DD, Chandran V. Treating psoriatic arthritis to target: defining Psoriatic Arthritis Disease Activity Score (PASDAS) that reflects state of minimal disease activity (MDA). J Rheumatol. 2019. https://doi.org/10.3899/jrheum.181472. 10. Vanier A, Mariette X, Tubach F, Fautrel B, Group SS. Cost-effectiveness of TNF-blocker injection spacing for patients with established rheumatoid arthritis in remission: an economic evaluation from the spacing of TNF- blocker injections in rheumatoid arthritis trial. Value Health. 2017;20(4): 577–85. 28. Helliwell PS, FitzGerald O, Fransen J, Gladman DD, Kreuger GG, Callis-Duffin K, et al. Author details 1 1Department of Rheumatology, Sint Maartenskliniek, Hengstdal 3, 6574 NA Nijmegen, The Netherlands. 2Department of Rheumatic Diseases, Radboud Institute of Health Sciences, Radboud University Medical Centre, Nijmegen, The Netherlands. 3Department of Rheumatology, Leiden University Medical Centre, Leiden, The Netherlands. Page 11 of 12 Page 11 of 12 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (20 Michielsens et al. 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An economic evaluation of stopping versus continuing tumor necrosis factor inhibitor treatment in rheumatoid arthritis patients with disease remission or low disease activity: results from a pragmatic open-label trial. Arthritis Rheumatol. 2018;70(10):1557–64. 31. Molto A, Gossec L, Meghnathi B, Landewe RBM, van der Heijde D, Atagunduz P, et al. An Assessment in SpondyloArthritis International Society (ASAS)-endorsed definition of clinically important worsening in axial spondyloarthritis based on ASDAS. Ann Rheum Dis. 2018;77(1):124–7. 13. Coates LC, Navarro-Coy N, Brown SR, Brown S, McParland L, Collier H, et al. The TICOPA protocol (tight control of psoriatic arthritis): a randomised controlled trial to compare intensive management versus standard care in early psoriatic arthritis. BMC Musculoskelet Disord. 2013;14:101. 32. Machado PM, Landewe R, Heijde DV. Assessment of SpondyloArthritis international S. Ankylosing Spondylitis Disease Activity Score (ASDAS): 2018 update of the nomenclature for disease activity states. Ann Rheum Dis. 2018;77(10):1539–40. 14. Edwards CJ, Fautrel B, Schulze-Koops H, Huizinga TWJ, Kruger K. Dosing down with biologic therapies: a systematic review and clinicians' perspective. Rheumatology (Oxford). 2017;56(11):1847–56. 33. Piaggio G, Elbourne DR, Pocock SJ, Evans SJ, Altman DG, Group C. Reporting of noninferiority and equivalence randomized trials: extension of the CONSORT 2010 statement. JAMA. 2012;308(24):2594–604. 15. Navarro-Compan V, Plasencia-Rodriguez C, de Miguel E, Balsa A, Martin- Mola E, Seoane-Mato D, et al. Anti-TNF discontinuation and tapering strategies in patients with axial spondyloarthritis: a systematic literature review. Rheumatology (Oxford). 2016;55(7):1188–94. 34. Rehal S, Morris TP, Fielding K, Carpenter JR, Phillips PP. Non-inferiority trials: are they inferior? A systematic review of reporting in major medical journals. BMJ Open. 2016;6(10):e012594. 16. Cantini F, Niccoli L, Cassara E, Kaloudi O, Nannini C. Received: 27 September 2019 Accepted: 16 December 2019 17. Gratacos J, Pontes C, Juanola X, Sanz J, Torres F, Avendano C, et al. Non- inferiority of dose reduction versus standard dosing of TNF-inhibitors in axial spondyloarthritis. Arthritis Res Ther. 2019;21(1):11. 18. Landewe R, Sieper J, Mease P, Inman RD, Lambert RG, Deodhar A, et al. Efficacy and safety of continuing versus withdrawing adalimumab therapy in maintaining remission in patients with non-radiographic axial spondyloarthritis (ABILITY-3): a multicentre, randomised, double-blind study. Lancet. 2018;392(10142):134–44. References Duration of remission after halving of the etanercept dose in patients with ankylosing spondylitis: a randomized, prospective, long-term, follow-up study. Biologics. 2013;7:1–6. 35. Fautrel B, Pham T, Alfaiate T, Gandjbakhch F, Foltz V, Morel J, et al. Step- down strategy of spacing TNF-blocker injections for established rheumatoid Page 12 of 12 Michielsens et al. Trials (2020) 21:90 Michielsens et al. Trials (2020) 21:90 arthritis in remission: results of the multicentre non-inferiority randomised open-label controlled trial (STRASS: Spacing of TNF-blocker injections in rheumatoid arthritis study). Ann Rheum Dis. 2016;75(1):59–67. arthritis in remission: results of the multicentre non-inferiority randomised open-label controlled trial (STRASS: Spacing of TNF-blocker injections in rheumatoid arthritis study). Ann Rheum Dis. 2016;75(1):59–67. 36. Campbell G. Guidance for the Use of Bayesian Statistics in Medical Device Clinical Trials. 2010. https://www.fda.gov/regulatory-information/search-fda- guidance-documents/guidance-use-bayesian-statistics-medical-device- clinical-trials/. Accessed 20 Aug 2019. 37. Sterne JA, White IR, Carlin JB, Spratt M, Royston P, Kenward MG, et al. Multiple imputation for missing data in epidemiological and clinical research: potential and pitfalls. BMJ. 2009;338:b2393. 38. Moons KG, Donders RA, Stijnen T, Harrell FE Jr. Using the outcome for imputation of missing predictor values was preferred. J Clin Epidemiol. 2006;59(10):1092–101. 39. Goekoop-Ruiterman YP, de Vries-Bouwstra JK, Allaart CF, van Zeben D, Kerstens PJ, Hazes JM, et al. Clinical and radiographic outcomes of four different treatment strategies in patients with early rheumatoid arthritis (the BeSt study): a randomized, controlled trial. Arthritis Rheum. 2008;58(2 Suppl): S126–35. 40. Heimans L, Akdemir G, Boer KV, Goekoop-Ruiterman YP, Molenaar ET, van Groenendael JH, et al. Two-year results of disease activity score (DAS)- remission-steered treatment strategies aiming at drug-free remission in early arthritis patients (the IMPROVED-study). Arthritis Res Ther. 2016;18:23. 41. Mottonen T, Hannonen P, Leirisalo-Repo M, Nissila M, Kautiainen H, Korpela M, et al. Comparison of combination therapy with single-drug therapy in early rheumatoid arthritis: a randomised trial. FIN-RACo trial group. Lancet. 1999;353(9164):1568–73. 42. Verschueren P, De Cock D, Corluy L, Joos R, Langenaken C, Taelman V, et al. Methotrexate in combination with other DMARDs is not superior to methotrexate alone for remission induction with moderate-to-high-dose glucocorticoid bridging in early rheumatoid arthritis after 16 weeks of treatment: the CareRA trial. Ann Rheum Dis. 2015;74(1):27–34. 43. de Jong PH, Hazes JM, Han HK, Huisman M, van Zeben D, van der Lubbe PA, et al. References Randomised comparison of initial triple DMARD therapy with methotrexate monotherapy in combination with low-dose glucocorticoid bridging therapy; 1-year data of the tREACH trial. Ann Rheum Dis. 2014; 73(7):1331–9. 44. van Lonkhuyzen L. Prijzenoorlog rond bestverkochte medicijn tegen reuma. NRC. 2018. https://www.nrc.nl/nieuws/2018/12/05/prijzenoorlog-rond- bestverkochte-medicijn-tegen-reuma-humira-a3059663/. Accessed 20 Aug 2019. 45. Medicijnkosten.Zorginstituut Nederland.2019. https://www.medicijnkosten. nl/. Accessed 20 August 2019. 45. Medicijnkosten.Zorginstituut Nederland.2019. https://www.medicijnkosten. nl/. Accessed 20 August 2019. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.	20,829
https://openalex.org/W1667409657_1	Spanish-Science-Pile	Open Science	Various open science	2,010	Diseño de una metodología para diagnosticar la situación actual de las Pymes en el Ecuador	None	Spanish	Spoken	7,344	13,821	Diseño de una metodologı́a para diagnosticar la situación actual de las Pymes en el Ecuador Nelson Alomoto Departamento de Ciencias Administrativas nelson.alomoto@epn.edu.ec Resumen La situación actual de nuestro paı́s, a la luz de diversos indicadores internacionales es una de las peores, a nivel latinoamericano y mundial. Para los gobernantes de turno, polı́ticos, empresarios, y en general para quienes están encargados de la toma de decisiones se ha tornado de vital importancia indicadores tales como los de: desarrollo humano, clima de negocios, competitividad, etc. Precisamente este último ha adquirido una importancia capital, de tal forma que la mayorı́a de las naciones han creado ministerios o dependencias gubernamentales relacionadas a la competitividad. Si bien la competitividad es un término que se lo cita en diferentes contextos, y de manera muy frecuente, es necesario no solo aclararla sino reconsiderarla a ﬁn de intentar medirla desde otra concepción acorde a los nuevos retos sociales de la época. Asimismo, la mayorı́a de los estudios e investigaciones respecto a competitividad hacen referencia a grandes empresas o naciones, y muy pocos son los esfuerzos que se han orientado hacia la competitividad de las Pequeñas y Medianas Empresas (Pymes), las que representan un importante sector de la economı́a no solo a nivel nacional sino a nivel mundial. En esta investigación se hace una aproximación conceptual hacia el constructo “competitividad”, desde la perspectiva de la teorı́a de los stakeholders o partes interesadas, intentando armonizar con los conceptos del bien común. El grado de informalidad latente en nuestro paı́s, lo que se evidencia en la falta de información y la escasa colaboración de los sectores público y privado, obstruyó la fase de implementación del presente proyecto, sin embargo, se propone una nueva deﬁnición de competitividad que permitirá evaluar el estado actual de las Pymes en nuestro paı́s. Palabras claves: Competitividad, pequeñas y medianas empresas, partes interesadas (o involucradas). Abstract The current situation of our country, in light of various international indicators is one of the worst, in Latin America and worldwide. For those in power, politicians, businessmen, and in general for those charged with decisionmaking has become of vital importance to indicators such as: human development, business climate, competitiveness, etc. Precisely the latter has gained importance, so that most countries have created ministries or government agencies related to competitiveness. While competitiveness is a term that refers to it in different contexts, and very often, it is necessary not only to clarify but to attempt to reconsider the measure from another design according to the new social challenges of the time. Also, most of the studies and research on competitiveness refer to corporations or nations, and few efforts have been geared towards competitiveness of Small and Medium Enterprises (SMEs), representing an important sector the economy not only nationally but worldwide. This research is a conceptual approach to the construct çompetitiveness”, from the perspective of stakeholder theory, trying to harmonize with the concepts of common good. The degree of informality latent in our country, as evidenced in the lack of information and lack of cooperation of public and private sectors, obstructed the implementation phase of this project, however, proposes a new deﬁnition of competitiveness that will allow assess the current status of SMEs in our country. Keywords: competitiveness, SMEs, stakeholders. 1 Introducción En los últimos tiempos, el concepto de asociatividad sur- tos por agrupar naciones con la ﬁnalidad de conseguir ge como un elemento vital en el desarrollo de los pue- ciertos objetivos que no se logran alcanzar de manera blos. A nivel mundial, se han realizado diversos inten- individual. Prueba de esto lo constituye la Unión Euro- 29 Nelson Alomoto pea, y en el caso de América Latina la UNASUR. Centroamérica cuenta con el SICA, Sistema de Integración Centroamericano. La Unión Europea no solo es una agrupación geográﬁca sino que constituye una estrategia multinacional para enfrentar los retos del nuevo milenio. La Estrategia de Lisboa, también conocida como Agenda de Lisboa es un plan de desarrollo que la Unión Europea se planteó el año 2000 con el objetivo estratégico de convertir a Europa en la economı́a del conocimiento más competitiva y dinámica del mundo, antes del 2010 [63]. En el ámbito latinoamericano existen iniciativas similares como la UNASUR (Unión de naciones sudamericanas), que se inició en el año 2008 con 12 paı́ses: Ecuador, Bolivia, Perú, Venezuela, Colombia, Paraguay, Chile, Argentina, Brasil y Uruguay, Surinam, y Guyana, y cuyo modelo de integración busca “favorecer un desarrollo más equitativo, armónico e integral de América del Sur” [11, p. 1]. Uno de los apartados del tratado constitutivo de la UNASUR, celebrado en Brasilia, el 23 de mayo de 2008, expresa que uno de sus objetivos es: “m) la integración industrial y productiva, con especial atención en las pequeñas y medianas empresas, las cooperativas, las redes y otras formas de organización productiva;” [57, p. 3]. En los últimos tiempos se realizan esfuerzos orientados a la integración de América latina y la Unión Europea [42]. Al margen de la evaluación de los resultados de estos proyectos multinacionales, y dentro de los lineamientos de la presente investigación, surgen elementos comunes para el análisis. La perspectiva de desarrollo conjunto de los paı́ses miembros, el interés por el incremento de la competitividad y la atención prioritaria a las pequeñas y medianas empresas cobran especial relevancia en los acuerdos mencionados. Las pequeñas y medianas empresas (Pymes), cada vez juegan un rol más protagónico en la economı́a mundial, y en especial, en economı́as menos desarrolladas y emergentes. Según la ONUDI, organización de las Naciones Unidas para el Desarrollo Industrial, alrededor del 90 % de las empresas en el mundo son Pymes y cerca del 60 % del empleo es generado por éstas [39, p. 1]. de empleo [44]. El estudio sobre responsabilidad social de la empresa, en las Pymes de Latinoamérica, auspiciado por el Banco Interamericano de Desarrollo, señala que el 98,3 % de empresas en Latinoamérica corresponden a Pymes, y resalta el hecho de que: “Resulta bien conocido el importante papel que las PyMEs juegan en la economı́a y la sociedad latinoamericana en términos de generadores de riqueza, empleo y cohesión social, donde este papel es extensible a todos los paı́ses que conforman la región. Por tanto, la mejora de la competitividad empresarial de este colectivo pasa por ser uno de los elementos claves para desarrollar tanto las economı́as como las sociedades latinoamericanas” [62, p. 1]. En el caso del Ecuador, las Pymes no han tomado relevancia desde la perspectiva de las polı́ticas de estado. Prueba de ello es que existe escasa información de base que permita conocer ciertos aspectos fundamentales sobre este sector. Dentro de los estudios aislados de aproximación sobre las Pymes; el Instituto Nacional de Estadı́stica y Censos (INEC), da cuenta que el 47 % de empresas manufactureras corresponden a la pequeña empresa y el 30 % a la mediana industria [31]. En el mismo informe se anota que, a pesar que existen mayor cantidad de pequeñas industrias, la producción total más importante corresponde a las industrias de grandes proporciones con un 90 % de la producción total. Esto demuestra la gran asimetrı́a que está presente en nuestro paı́s. Un informe más reciente proporcionado por el Ministerio de Industrias y Competitividad en el año 2008, determina que el 13,3 % corresponde a la gran industria, el 19,9 % a la mediana industria, el 43,2 % a la pequeña industria, y el 23,6 % a la micro industria. El mismo informe advierte sobre el grado de conﬁabilidad del mismo debido al nivel de informalidad de sector micro industrial [37]. En la literatura especializada no se encuentran muchos trabajos sobre la gestión en pequeñas y medianas empresas, y mucho más escasas y recientes son sobre su competitividad; tradicionalmente los esfuerzos, teorı́as, herramientas, metodologı́as, estudios, estadı́stigrancas, etc., han sido orientados hacia organizaciones granempresa pymes des, y adaptadas a las Pymes, a pesar de que existen 10% 90% sustanciales diferencias entre empresas grandes y Pymes [41]. “Aun si los modelos generales fueran aplicados correctamente, deberı́an ser inadecuados para las caracterı́sticas particulares de las Pymes: la pequeña empresa es diferente de la gran compañı́a; usted no puede simplemente mirar las necesidades de las Pymes poniendo sus binoculares de arriba hacia abajo y volviendo pequeño lo que era grande” [23, p. 28]. La situación actual de nuestro paı́s, a la luz de diverFigura 1. Porcentaje a nivel mundial. Fuente: Onudi,2008 sos indicadores internacionales es una de las peores, a En Europa representan más del 99 % de los negocios nivel latinoamericano y mundial. Uno de los indicado[41], y especı́ﬁcamente en España el 99,87 % [5]. En Co- res que ubican al Ecuador en los niveles más bajos es lombia las micro, pequeñas y medianas empresas cons- el de competitividad. En el ranking 2009-2010 del Foro tituyen el 99 % de los establecimientos, y generan el 70 % Económico Mundial ocupamos el puesto 105 entre 133 30 Revista Politécnica, 2010, Vol. 29(1): 29–41 Diseño de una metodologı́a para diagnosticar la situación actual de las Pymes . .. paı́ses [52, p. 13], mientras que de acuerdo al ı́ndice de rendimiento industrial competitivo de la ONUDI, ocupamos el puesto 112 entre 122 paı́ses [40, pp. 28 – 30], lo que podrı́a llevar a la conclusión de que nuestra industria y nuestras Pymes se encuentran en un estado de desarrollo incipiente, lo que preocupa tanto a empresarios, gobernantes, y también deberı́a activar las alarmas en el sector educativo, especialmente el universitario, donde se forman los cuadros productivos de una sociedad. Si bien la competitividad es un término que se lo cita en diferentes contextos, y de manera muy frecuente, es necesario no solo aclararla sino reconsiderarla a ﬁn de intentar medirla desde otra concepción acorde a los nuevos retos sociales de la época. Asimismo, la mayorı́a de los estudios e investigaciones respecto a competitividad hacen referencia a grandes empresas o naciones, y muy pocos son los esfuerzos que se han orientado hacia la competitividad de las Pequeñas y Medianas Empresas Pymes, las que representan un importante sector de la economı́a no solo a nivel nacional sino a nivel mundial. Surgen entonces las siguientes interrogantes que se intentará responder en este estudio: • ¿Qué es la competitividad? su competitividad, se la obtiene proyectando informes internacionales, que sobre competitividad han levantado organismos internacionales sobre la gran industria, o algún estudio comparativo que se ha realizado aplicando acrı́ticamente modelos de otros paı́ses, sin considerar las caracterı́sticas especı́ﬁcas de nuestra realidad. Por su lado, la universidad ecuatoriana, no ha logrado cumplir con la misión que le corresponde dentro la sociedad, en lo que concierne a la formación del talento humano capacitado que requiere el sector productivo. Sus aulas se han convertido en el escenario perfecto para transmitir experiencias exógenas, rellenando a sus alumnos de herramientas gerenciales de moda, generalmente diseñadas para las grandes empresas, a pesar de que los emprendedores que salen de la universidad y arrancan su idea de negocio, generalmente inician con una Pyme. Al momento de diseñar nuevas carreras universitarias, no se toman en consideración las necesidades de la sociedad, por lo que son incompatibles con el desarrollo socioeconómico nacional [17]. Persiste el divorcio de la empresa-universidad, so pretexto de privacidad en un caso, y autonomı́a en el otro, cumpliéndose la conjetura de Pérez y Peiró: • ¿Cómo se la mide? “La universidad sólo se legitima si responde a las de• ¿Cuáles son las consecuencias de la competitivi- mandas y necesidades sociales para las que ha sido creadad? da y que justiﬁcan su existencia continuada y su dimen• Según la concepción actual, ¿cuál es el nivel com- sión social. Si se pierden o se anulan los sensores de la universidad ante las demandas y necesidades sociales, la petitivo de nuestro paı́s? toma de decisiones comienza a estar determinada bási• Es necesario reconceptualizar el término competi- camente por el juego polı́tico interno y por una lógica de tividad, ¿de qué forma? intereses corporativistas y de juegos de poder entre los diferentes grupos y estamentos de la propia institución”. • ¿Cómo medirla? Citado en [22, p. 2]. Para intentar responder a estas interrogantes, la presente investigación se plantea: El presente estudio asumió como un hecho indiscutible la colaboración de entidades públicas y privadas pa1. Elaborar un marco teórico que permita establecer ra la captura y procesamiento de datos, sin embargo, en el estado del arte respecto a la competitividad, y la práctica se evidenció la poca, escasa y en ocasiones sus formas de medirla. nula colaboración de éstas para conﬁar datos, peor aún 2. Determinar el estado actual de competitividad de información sobre su gestión o de terceros. Es ası́ que nuestra industria. la propuesta inicial de la presente investigación incluı́a la implementación de una metodologı́a para diagnosti3. Proponer una nueva deﬁnición de competitividad. car la situación actual de las Pymes en el sector norte 4. Aplicar la nueva medición de competitividad a las del paı́s, sin embargo, la negativa explicita del Servicio de Rentas Internas SRI, cerró deﬁnitivamente la posibiliPymes del sector centro-norte del paı́s. dad de acceder al universo de la Pymes para efectuar el La primera gran diﬁcultad con la que se encuentra muestreo proyectado. En ausencia de un muestreo adecualquier investigador en el Ecuador, es que existen es- cuado, cualquier estudio carece de validez estadı́stica y casas fuentes sistematizadas de información, y las que sus conclusiones no son representativas. Consecuenteexisten, repiten información poco relevante. De otro la- mente, se tomó la decisión de omitir la fase de impledo, como ya se ha mencionado, las Pymes juegan un pa- mentación, y en su lugar, profundizar en el análisis teóripel importante en la economı́a del paı́s, sin embargo, ni co de un concepto: ”peligroso, obsesivo, elusivo o sin a nivel público ni privado existen estudios dirigidos a signiﬁcado”[3, p. 63], que ha tenido numerosas interpreconocer la situación actual por la que atraviesan. La po- taciones y tiende a ser ambiguo, como es la competitivica información disponible, como por ejemplo respecto a dad [54]. Revista Politécnica, 2010, Vol. 29(1): 29–41 31 Nelson Alomoto 2 Marco teórico 2.2 El presente estudio utiliza el método cualitativo de investigación, apropiado para las Ciencias Administrativas; empieza con una revisión documental histórica bastante amplia, que no pretende ser completa ni exhaustiva, de cómo el concepto de competitividad ha venido utilizándose en diferentes contextos, a ﬁn de establecer el estado del arte en esta dirección. Se aborda la discusión del término competitividad desde distintos enfoques: macroeconómico y microeconómico, intentando relacionar con las consecuencias o el impacto que esta concepción tradicional de competitividad ha tenido sobre el entorno y los involucrados de la gestión empresarial. Posteriormente se desarrolla el marco en el cual se hace la propuesta de la nueva deﬁnición de competitividad, que inevitablemente conduce a empatar con una teorı́a que todavı́a está en discusión, como es la teorı́a de los stakeholders. Finalmente se propone una forma de deﬁnir un indicador de competitividad, acorde a la nueva deﬁnición propuesta. Adicionalmente se precisan ciertos aspectos que deberán considerarse para una posterior fase de aplicación de este instrumento de medición, en los ocho sectores especı́ﬁcos de las Pymes. 2.1 Competitividad La competitividad es un concepto muy polémico, todos hablan de competitividad y pocos saben exactamente lo que signiﬁca [51], ha merecido muchos esfuerzos de investigación, diversas concepciones e interpretaciones [9], [46], [54] y sobre el que se ha llegado a aﬁrmar que es una “peligrosa obsesión” [33]. Si bien las discusiones y aportes teóricos se orientan en su mayorı́a a la competitividad de naciones y a sectores industriales de grandes empresas, en los últimos tiempos se han desarrollado varias investigaciones sobre la competitividad en micro, pequeñas y medianas empresas [50], [10], [19], [45], [8], [29]. Al igual que la productividad, la competitividad en las pequeñas y medianas empresas a nivel mundial ha constituido el foco de atención de los gobiernos, al atraer importantes esfuerzos de promoción, fomento y asistencia técnica [58]. Para entender los diferentes aspectos que involucra el término competitividad, y consecuentemente su medición, a continuación se revisa algunos aspectos teóricos relacionados. Una primera clasiﬁcación sugiere que hay tres niveles de análisis en el estudio de la competitividad: el marco económico general, el sector industrial y la propia empresa [60], en la misma dirección otros autores la clasiﬁcan en: estructural, sectorial y empresarial [24]. En el primer nivel se habla de competitividad de las naciones, en el siguiente de regiones o sectores productivos, mientras que en último se reﬁere a la competitividad de la empresa. 32 Enfoque macroeconómico Históricamente el concepto de competitividad ha sido enfocado desde diversos puntos de vista; en el cuadro 1 se reproducen algunas deﬁniciones que se han dado a través del tiempo y que son citadas por Alonso y Barcenilla, con la ﬁnalidad de establecer un análisis crı́tico de las mismas. Aunque es explı́cito solo en la deﬁnición de la Comisión de EE.UU. de 1985, en todos los enfoques la noción macroeconómica de competitividad es consustancial a economı́as de libre mercado, es decir, sin el control directo del estado. En su primer enfoque predomina como objetivo central y quizá único, la preocupación de la balanza comercial de un paı́s. Evidentemente, acciones estatales tales como subsidios a ciertos ı́tems exportables, barreras arancelarias a productos provenientes del exterior, ası́ como devaluaciones monetarias en economı́as no dolarizadas, tienden a manipular artiﬁcialmente los saldos comerciales, lo que pone de relieve la inconsistencia de esta perspectiva. El objetivo central, en el segundo enfoque, es mantener una balanza comercial positiva, además se agregan y relacionan con los objetivos sobre crecimiento de la renta y el empleo. La sostenibilidad es el agregado que pretende dar un toque dinámico al tercer enfoque. Ahora bien, si entendemos por sostenibilidad la caracterı́stica o estado según el cual pueden satisfacerse las necesidades de la población actual y local sin comprometer la capacidad de generaciones futuras, surge entonces la pregunta: ¿será suﬁciente pedir sostenibilidad en un mundo donde las condiciones cambian abruptamente?. Precisamente el colapso inmobiliario que acabamos de presenciar en el 2008 supone haber dado un paso hacia una discontinuidad económica. En este sentido la sostenibilidad es inherentemente estática, por lo que se hace necesario buscar nuevos paradigmas. Posiblemente la elasticidad sea la respuesta ante estos nuevos escenarios turbulentos en donde la incertidumbre es lo único seguro. A diferencia de la sostenibilidad que pretende alcanzar un punto óptimo de estabilidad, la elasticidad propone convivir con la posibilidad del colapso y evolucionar con él; mientras que la sostenibilidad es un estado quebradizo, es supervivencia, la elasticidad implica prosperar siendo capaces de superar lo inesperado. [15]. El cuarto enfoque sugiere que la productividad es la condición necesaria para la competitividad de un paı́s. Asimismo, supone que la competitividad de un paı́s implica un alto nivel de vida para sus habitantes y lo contrario para sus paı́ses competidores, en un juego ganar– perder. La obsesión peligrosa a la que hace referencia Krugman posiblemente ha llevado a nuestro mundo industrializado a relegar objetivos sociales y ambientales a favor de la productividad [32], o lo que De Sousa Santos llama la mono cultura del productivismo capitalista, según la cual el crecimiento económico y la productividad medida en un ciclo de producción determinan la productividad del trabajo humano o de la naturaleza [20]. En efecto, las consecuencias ambientales de la de- Revista Politécnica, 2010, Vol. 29(1): 29–41 Diseño de una metodologı́a para diagnosticar la situación actual de las Pymes . .. senfrenada carrera por la productividad–competitividad se pueden observar en la alteración climática del globo, lo que nos ha llevado a superar las 350 ppm de C O2 en la atmosfera, que es el lı́mite determinado para llegar a la emergencia climática [1]. El 70 % de la polución global corresponde a las Pymes, ası́ como el 60 % de las emisiones totales de carbón, y el total de suma de los impactos ambientales de las Pymes pesa más que el impacto ambiental combinado de ﬁrmas grandes [29]. Tabla 1. Deﬁniciones de Competitividad. Fuente: Alonso & Barcenilla, 1999, págs. 22-23 ENFOQUE 1. Resultados del comercio 2. Relación balanza de pagos–crecimiento 3.Resultado exterior compatible con mejora del nivel de renta per cápita 4. Vı́nculo con productividad DEFINICIÓN “La competitividad internacional alude a la capacidad de los productores de un paı́s para competir con éxito tanto en los mercados mundiales como con respecto a las importaciones en su propio mercado doméstico (. . . ) La competitividad en este sentido muy general viene a ser sinónimo del comportamiento económico agregado” (His Majesty’s Treasury, UK, 1983, p.1). “La aptitud de un paı́s (o grupo de paı́ses) para enfrentar la competencia a nivel mundial: considerando tanto su capacidad para exportar y vender en los mercados externos como para defender el mercado doméstico de una excesiva penetración de las importaciones” (Chesnais, 1988, p.8). “La capacidad de un paı́s para alcanzar los objetivos centrales de la polı́tica económica, especialmente el crecimiento de la renta y del empleo, sin recurrir en diﬁcultades de balanza de pagos” (Fagerberg, 1988, p.355) “La capacidad para hacer retroceder los lı́mites de la restricción externa al crecimiento” (Mathis, Mazier y Rivaud-Danset, 1988, p.7). “La habilidad para (i) vender muchos productos y servicios (para satisfacer una restricción externa): (ii) en factor ingresos acordes con (actuales y cambiantes) niveles de aspiración de los paı́ses; y (iii) en condiciones-macro del sistemas social, ambiental, económico sean satisfactorios a la población” (Aiginger, 1998) “La competitividad es la capacidad de un paı́s para sostener y expandir su cuota en el mercado internacional y al mismo tiempo incrementar el nivel de vida de la gente” (Fajnzylberg, 1988, p.12). “Competitividad es el grado en el que una nación puede, bajo condiciones de mercado libre, producir bienes y servicios que superen la prueba de los mercados internacionales, mientras simultáneamente mantiene o expande la renta real de sus ciudadanos” (Comisión Presidencial sobre Competitividad Industrial de EE.UU.,1985). “La habilidad de una economı́a para proveer a su población de altos y crecientes estándares de vida y altas tasas de empleo sobre una base sostenible” (Comisión Europea, 2001). “Desarrollo de una superior eﬁciencia y con la capacidad de una economı́a para incrementar el producto de las actividades de más alta productividad, que, a su vez, pueden generar elevados niveles de salarios en términos reales” (Cohen, Tysson, Teece, y Zysman, 1984, p.2). “La capacidad de un paı́s para asegurar un alto nivel de vida de sus ciudadanos relativo a los ciudadanos de otros paı́ses, tanto en el presente como en el futuro” (Fagerberg, 1996, p.48). “Ya nadie duda que el rápido y creciente deterioro global de los ecosistemas es claramente antropogénico. Sin embargo, el reparto de responsabilidad no es homogéneo. Es el sistema productivista, basado en el consumo creciente y en la velocidad, e impuesto por los paı́ses ricos a través de la denominada globalización, el principal responsable de la destrucción” [28, p. 152]. petitividad, su medición aparece aún más polémica. Existen actualmente dos clases de indicadores macroeconómicos de competitividad, que son muy populares al tiempo que controversiales; corresponden a organizaciones que se encargan de la elaboración y publicación de rankings anuales de competitividad mundial y son: WCY World Competitiveness Yearbok del Institute for A nombre de productividad se ha llegado a extremos Management and Development (IMD), y el Global Cominsospechados de explotación del recurso humano, “al petitiveness Report (GCR), del Foro Económico Mundial punto de haber logrado en el presente la más elevada (WEF). La deﬁnición de competitividad del IMD maniproductividad nunca antes alcanzada por la humanidad; ﬁesta: paradójicamente, también ha logrado una creciente po“Como las naciones y los negocios manejan la totalibreza de más de 1.200 millones de pobres y hambrientos dad de sus capacidades para alcanzar una gran prosperien cada vez más espacios de nuestra aldea mundial” [38, dad”. La competitividad no es crecimiento o resultados p. 16]. económicos, pero deberı́a tener en cuenta “los factores Al no existir un acuerdo sobre el concepto de com- suaves” de competitividad, tales como el ambiente, la Revista Politécnica, 2010, Vol. 29(1): 29–41 33 Nelson Alomoto calidad de vida, tecnologı́a, conocimiento, etc. [30, p. 2]. El reporte del Foro Económico Mundial (WEF), versa sobre 133 economı́as, recoge información de 100 indicadores, y se soporta en 12 pilares (citados en el documento completo). El Foro Económico Mundial deﬁne la competitividad de la siguiente manera: “Nosotros deﬁnimos la competitividad como el conjunto de de instituciones, polı́ticas y factores que determinan el nivel de productividad de un paı́s. El nivel de productividad, a su turno, establece el nivel sostenible de prosperidad que puede ser alcanzada por una economı́a. En otras palabras, economı́as más competitivas tienden a ser capaces de producir altos niveles de ingresos para sus ciudadanos. El nivel de productividad también determina las tasas de retorno obtenidas por inversionistas en una economı́a. Debido a que las tasas de retorno son los controladores fundamentales de las tasas de crecimiento de la economı́a, una economı́a más competitiva es aquella que probablemente crece lo más rápido del mediano a largo plazo” [52, p. 4]. Como se puede apreciar en el informe 2010, bajo esta métrica, Ecuador ocupa el puesto 105 entre 133 paı́ses, y a nivel Latinoamericano supera solamente a paı́ses como: Bolivia, Paraguay y Venezuela. En los últimos tiempos, la responsabilidad social corporativa, el desarrollo sustentable, y el respeto al medio ambiente han ganado terreno en la discusión de la función social de la empresa; muestra de esto se evidencia en el Índice de Competitividad Responsable, un informe que se viene publicando desde el 2002, y que pretende evaluar prácticas responsables empresariales en 108 paı́ses. Este nuevo punto de vista muestra lo relativo del concepto competitividad, pues si bien es cierto en este nuevo ranking no están los mismos paı́ses, no es menos cierto que la ubicación relativa de algunos paı́ses varı́a notablemente. Ecuador ocupa el puesto 79 en el Índice de Competitividad Responsable 2007 [65]. Todas las concepciones implı́citas en las deﬁniciones mencionadas, han conducido a la humanidad a una desenfrenada carrera por alcanzar ı́ndices o indicadores de competitividad. Estas concepciones ganar-perder suponen que la competitividad de un paı́s determinan la menor competitividad de otros; la obsesión por la productividad ha dado lugar a niveles inimaginables de explotación de la mano de obra y la sobre utilización de los recursos no renovables, con las consecuencias negativas sobre el ambiente que nos rodea y las asimetrı́as sociales en el planeta. “La competitividad irresponsable tiene consecuencias catastróﬁcas. Ya estamos sintiendo los impactos tempranos del cambio climático, tales como el derretimiento de los mantos glaciares y la diﬁcultad para conseguir el sustento de millones de personas. Los mercados y la economı́a polı́tica más amplia de hoy dı́a se ven cada vez más como cómplices de las crecientes desigualdades entre y dentro de las naciones, la escasez crónica de agua, la emigración caótica, la corrupción incesante y el creciente nacionalismo económico” [65, p. 12]. Para abandonar las obsesivas ideas actuales de com34 petitividad, Aiginger propone llegar a un acuerdo general para deﬁnir la competitividad como “la capacidad para crear el bienestar” [3], y de esta manera corregir los defectos que las deﬁniciones y concepciones anteriores traen consigo. Es más, propone evaluar el proceso y el resultado. La competitividad resultante serı́a medida por una función de bienestar W, deﬁnida como: W = f (Y, S, E), sujeto a FS, EBS, PS, LEIS. Es decir, supone la maximización de la función de bienestar W, que a su vez depende de las variables: Y, ingreso per cápita, S, conjunto de indicadores sociales y distribucionales, y E, conjunto de indicadores ecológicos; sujeto a un conjunto de restricciones dados por: FS, Sostenibilidad ﬁnanciera, EBS, sostenibilidad de balances externos, PS, estabilidad polı́tica y LEIS, ocio. Es una propuesta que, por un lado busca eliminar las falencias analizadas en las concepciones tradicionales y por otro incorporar elementos que son reclamados por las nuevas ideas de la función social empresarial tales como: respeto al medio ambiente, sostenibilidad, etc. 2.3 Enfoque microeconómico Contrariamente a lo que se podrı́a pensar, el término competitividad a nivel microeconómico no está bien deﬁnido [3], peor aún aceptado. A nivel macroeconómico, la competitividad de las naciones no es una cualidad propia de los estados, viene representada por las cualidades de sus organismos, empresas y productos al competir en mercados locales e internacionales [24]. Existen muchas formas de medir y deﬁnir la competitividad a nivel de empresa, éstas se basan cuantitativamente en la participación de mercado, indicadores de productividad y/costo, márgenes de ganancia y/o beneﬁcios netos. Al igual que en el nivel macro, existen mediciones y deﬁniciones de tipo cualitativo como la investigación y desarrollo además de estrategias gerenciales [34]. La mayorı́a de formas de medir la competitividad, sino todas, tienen algo en común; terminan en un ranking comparativo difı́cil de implementar en el ámbito de las pequeñas y medianas empresas [53]. En este sentido, cobra vital importancia precisar el concepto de competitividad a nivel empresarial. A nivel de empresas e industrias, el concepto está vinculado directamente con la habilidad de éstas para operar rentablemente en un mercado determinado [43], o la capacidad para ser rentable de un modo sostenible en una economı́a libre de proteccionismos [16]. En esta concepción la rentabilidad es un objetivo, quizá el único del accionista, pero, la empresa es una institución social, por tanto, su razón de ser es social, es decir, que el objetivo de una empresa no se puede reducir solo a ganar dinero. Este es un requisito imprescindible, pero no suﬁciente [6]. “En una encuesta realizada por la empresa McKinsey a 4.238 ejecutivos en 116 paı́ses, tan sólo uno de cada seis ejecutivos concuerda con la clásica posición Revista Politécnica, 2010, Vol. 29(1): 29–41 Diseño de una metodologı́a para diagnosticar la situación actual de las Pymes . .. de Milton Friedman, según la cuál la única responsabilidad de la empresa es maximizar sus utilidades.” [26, p. 24]. Si a nivel macro se acepta la propuesta de Aiginger para deﬁnir la competitividad de un paı́s como la capacidad de una nación para crear bienestar, a nivel microeconómico, la competitividad empresarial deberı́a apuntar en aquella dirección. De igual forma, a nivel empresarial, una adecuada evaluación deberı́a contener una evaluación del resultado (competitividad resultante) y una evaluación del proceso (proceso de competitividad). Consecuentemente, una deﬁnición razonable de competitividad empresarial deberı́a considerar la capacidad de una empresa para crear bienestar. Cada nación tiene la potestad de deﬁnir el concepto de bienestar, es decir no se puede pretender una deﬁnición universal de bienestar de los pueblos. Las deﬁniciones tradicionales de competitividad basadas en costos y balanzas de pagos conllevan implı́citamente la no competitividad de otras naciones, sin embargo la deﬁnición de competitividad concebida en la capacidad de crear bienestar excluye la posibilidad de conseguir este objetivo a expensas del no bienestar de otros. De igual forma, a nivel de empresa el bienestar debe considerar a todos sus involucrados en el entorno próximo donde se desarrolla ésta. El entorno comprende el conjunto de valores naturales, sociales y culturales existentes en un lugar y un momento determinado, que inﬂuyen en la vida del ser humano y en las generaciones venideras. Es decir, no se trata sólo del espacio en el que se desarrolla la vida sino que también abarca seres vivos, objetos, agua, suelo, aire y las relaciones entre ellos, ası́ como elementos tan intangibles como la cultura [36]. Entre los involucrados se encuentran: accionistas, empleados, clientes, proveedores, acreedores; es decir, todos aquellos que se ven afectados directa o indirectamente por la gestión empresarial. Existen más involucrados o afectados en el radio de acción de la empresa; la sociedad y el estado o gobiernos locales, por ejemplo. El estado o gobierno local que recauda los impuestos generados por la empresa y los revierte vı́a infraestructura y servicios generales a la sociedad. Los competidores están involucrados de manera directa en la gestión de la empresa, en tanto compiten por un mismo mercado. Los complementadores son otro tipo de involucrados en la gestión organizacional, en tanto la demanda de sus productos o servicios se ve alentada y complementada por aquellos generados por la ﬁrma. Este nuevo punto de vista sobre la empresa, en principio, no distingue el tipo de interés ni el tipo de inﬂuencia que tiene cada una de las partes sobre la empresa, lo cual evidentemente variará según el sector industrial y las condiciones locales de cada región. Surge entonces a la necesidad de articular este desarrollo con la teorı́a de involucrados o stakeholders, que a decir de Freeman son cualquier grupo o individuo que puede afectar o ser afectado por la consecución de los objetivos de la empresa [21]. Históricamente la competitividad de la empresa ha sido enfocada desde la perspectiva de la rentabilidad de los accionistas, es decir, solamente atendiendo a uno solo de sus involucrados, cuando en la práctica existen otros actores que invierten activos tangibles e intangibles como por ejemplo: el estado que invierte en infraestructura (vı́as, energı́a, etc.), los empleados (conocimiento, capacidad, etc.), los acreedores (dinero), y otros como las comunidades y el ambiente que en deﬁnitiva se ven afectados por la gestión empresarial. El aporte de la presente investigación radica en intentar deﬁnir la competitividad desde una óptica menos individualista que tome en cuenta a otros involucrados, es decir, los stakeholders. La teorı́a de stakeholders permite abordar un nuevo paradigma empresarial, donde aparece una concepción de empresa plural, donde la empresa “no es cosa de uno (accionista o propietario), ni exclusivamente de dos (propietarios y trabajadores), si no que la empresa debe ser entendida desde la pluralidad de “agentes”—los que afectan—que intervienen en ella y, por tanto la hacen posible, ası́ como desde todos aquellos “pacientes”— los que son afectados—por la organización empresarial” [25, pp. 208-209]. 3 Propuesta de deﬁnición de competitividad Como se ha expuesto, la competitividad, no es un concepto totalmente deﬁnido, de manera que sea universalmente aceptado, pues a decir de Porter, resultarı́a un intento fútil; no obstante, es necesario abordarla desde una perspectiva más amplia que tome en consideración al amplio espectro de involucrados, afectados y beneﬁciados, ası́ como la nueva función que debe cumplir la empresa en la sociedad. Evidentemente, todos los involucrados tienen diferente grado de interés y no se ven afectados de la misma forma, de igual forma, para las organizaciones, no todos los stakeholders tienen la misma importancia; si se trata de caliﬁcar: el producto, lo más importante son los clientes, si es la seguridad en el trabajo, los empleados, y cuando se trata de problemas ambientales, lo más importante es la comunidad o el gobierno [56]. Existen varias perspectivas desde las que se puede abordar la relación empresa-stakeholder. La perspectiva corporativa, la perspectiva stakeholder y la perspectiva conceptual. La perspectiva corporativa aborda como las corporaciones tratan con sus stakeholders, la perspectiva stakeholder investiga de que manera los stakeholders tratan de inﬂuenciar a las ﬁrmas, y ﬁnalmente, la perspectiva conceptual estudia las relaciones entre conceptos especı́ﬁcos tales como el bien común o desarrollo sostenible y las interacciones negocio–stakeholders [55]. Pero bien, quienes son los stakeholders, involucrados o concernientes? Al respecto, Carbonell aﬁrma que “la teorı́a de stakeholders supone una visión integral de las organizaciones, al pretender conseguir un equilibrio entre los grupos de interés de la empresa (clientes, accionistas, proveedores, empleados y sociedad)” [13, p. 69]. Con los Revista Politécnica, 2010, Vol. 29(1): 29–41 35 Nelson Alomoto antecedentes expuestos, a nivel empresarial, se propone la siguiente deﬁnición de competitividad: La competitividad de una empresa es la habilidad para crear el bienestar de sus stakeholders, contribuyendo de manera protagónica al bien común en el entorno donde realiza sus actividades. En términos operacionales, esto supone no solo la eﬁciencia productiva inherente a su negocio, sino también la eﬁciencia social de su accionar. Subyace también una nueva concepción de empresa. Parece que es tiempo de redeﬁnir el concepto de empresa, posiblemente en el sentido que lo menciona Yacuzzi al citar que: “La ﬁrma es un sistema de stakeholders que opera dentro de un sistema mayor de la sociedad anﬁtriona, que provee la infraestructura legal y de mercado necesaria para las actividades de las ﬁrmas” [64]. Algunas de las bondades que presenta esta propuesta de deﬁnición de competitividad: útiles cuando se utilizan muestras de Pymes [35], se propone utilizar fuentes subjetivas y tanto variables cualitativas como cuantitativas. El marco conceptual para medir el desempeño corporativo se basa en la teorı́a de Stakeholders, contrastado con los objetivos empresariales. Las variables de desempeño corporativo (ex-post), deben estar relacionadas con la satisfacción de sus stakeholders, entre esos participantes encontramos a: clientes, accionistas, proveedores, acreedores, empleados, gobiernos y sociedad, [59]. El indicador de competitividad debe estar ı́ntimamente relacionado con la deﬁnición conceptual que se adopta, y debe cumplir ciertas caracterı́sticas mı́nimas deseables. Estudios relacionados proponen ciertas caracterı́sticas para los indicadores tales como: la relevancia, precisión y disponibilidad [47]. En lo que corresponde a la ﬁabilidad, la mayor parte de los indicadores se obtienen a través de encuestas, explotación de registros administrativos o censos. En el 1. Es un concepto integrador, que considera a las partes caso de las encuestas, se pueden citar aquellas referidas a interesadas y afectadas por la gestión de la empresa, hechos, opiniones o actitudes, motivaciones o sentimiende forma tal que aspectos como la Responsabilidad tos [61]: Social Corporativa, quedan inmersos en él, y no necesitarı́an desarrollarse de forma ex post, a ﬁn de re- 1. Encuestas basadas en hechos: en las que los encuestados son preguntados por un aspecto cuantiﬁcable obmediar de alguna forma los impactos negativos de la jetivamente. Es decir, encuestas en las que cualquiera gestión empresarial. que fuera el encuestado habrı́a de dar una respuesta 2. La competitividad de una empresa, no necesariamensimilar. te implicarı́a la no competitividad de las otras empresas. En otras palabras, es un concepto ganar–ganar, a 2. Encuestas basadas en acciones: en las que los encuestados son preguntados acerca de si han llevado a cadiferencia de las deﬁniciones tradicionales, en donde bo o no una determinada acción en un momento del la competitividad de una empresa se logra a expensas tiempo. de las otras, en una concepción ganar-perder. Consecuentemente, conceptos como la solidaridad, la aso- 3. Encuestas de opinión: en las que los encuestados son preguntados acerca de su valoración subjetiva sobre ciatividad y la complementariedad, adquirirı́an releciertos acontecimientos. vancia dentro de una nueva economı́a solidaria. 3. La productividad dejarı́a de ser la condición necesa- En lo que respecta a la comparabilidad, los indicadores ria para la competitividad, pues serı́a desplazada por sirven para cotejar; en este sentido deben ser estandarila de eco eﬁciencia. zados de tal forma a que se puedan aplicar en otras re4. La competitividad dejarı́a de ser una obsesión peli- giones. Finalmente, la disponibilidad se reﬁere al grado de accesibilidad tanto temporal como territorial. grosa y se convertirı́a en una aspiración social. Estas son caracterı́sticas deseables para el conjunto De hecho, existen aspectos que deberı́an ser aﬁnados de indicadores, sin embargo la realidad del medio donde en futuras investigaciones, tal es el caso del concepto del se desarrolló esta investigación no permitió garantizar bien común. A este respecto, çada sociedad humana tie- los requisitos anotados. Según el diseño original del esne su propio bien común. Su contenido será distinto para tudio, el mismo iniciaba con la extracción de una muesuna familia, una empresa, un sindicato, la comunidad lo- tra representativa de las Pymes extraı́da de las bases del cal, una nación o el conjunto de la sociedad humana”[7, Servicio de Rentas Internas del Ecuador SRI, no obstanp. 6]. En nuestro paı́s, lo más aproximado al bien común te este organismo imposibilitó el acceso a sus bases de es el principio del buen vivir que consta en el Plan Es- datos, argumentando aspectos legales, según se aprecia en el Anexo 1 (documento completo). Por otro lado, si tratégico del Gobierno para el periodo 2009-2013. bien se pudo haber planteado indicadores con alto nivel de relevancia con información objetiva, como es el caso de los indicadores ﬁnancieros de Dupont, habrı́a si4 Propuesta de medición do en detrimento del grado de precisión. A este respecto, Acompañada a la deﬁnición de competitividad debe ir la existen estudios que no aconsejan utilizar la información forma de medirla. Debido a que las empresas son muy ﬁnanciera que emiten las Pymes, pues están altamente reticentes a aportar datos de tipo cuantitativo–objetivo distorsionadas [35], y no son conﬁables. Adicionalmen[18], y que las fuentes subjetivas de desempeño son más te, en nuestro medio es prácticamente imposible recabar 36 Revista Politécnica, 2010, Vol. 29(1): 29–41 Diseño de una metodologı́a para diagnosticar la situación actual de las Pymes . .. información de la gestión empresarial, ya que los direc- naremos U de la siguiente manera2 : tivos muestran total desconﬁanza en organismos estata7 les y centros de investigación universitarios1 . Sobre la U = ∑ pi ui realidad argumentada, se presenta el dilema de utilizar 1 información objetiva o subjetiva. Por lo expuesto se sugiere utilizar información de tipo subjetiva [19], [48], con con 0 ≤ pi ≤ 1, ∑ pi = 1, ui (mejor nivel) = 1 y base en encuestas dirigidas a los directivos de las Pymes. ui (peor nivel) = 0. Donde pi es el peso del stakeholder i y ui es el nivel del atributo i. Valores que se asignan a los componentes o stakeholders por parte del experto. Los diferentes stakeholders se indexan de la manera indicada en la tabla 2: 4.1 Dimensiones del indicador Tabla 2. Stakeholders de las empresas. El número de dimensiones incluido en la medida de Stakeholders Valor i competitividad es una reﬂexión de la complejidad del concepto, pero es también fuente de ambigüedad [54]. Accionistas 1 En la literatura especializada, se encuentran varias proEmpleados 2 puestas de indicadores que miden la competitividad de Clientes 3 las Pymes atendiendo a diferentes concepciones: [14], [45], [49], [53]. Al ser el concepto de competitividad comAcreedores 4 plejo en sı́ mismo, abarca diversas dimensiones, frente Proveedores 5 a lo que se proponen indicadores que suman o promeGobiernos 6 dian el valor de varias variables. Aún más, en algunos casos se evidencian mezclas de indicadores ex ante con Sociedad 7 ex post en un solo indicador ﬁnal que pretende clasiﬁcar en un ranking de empresas competitivas vs las que no En primer lugar, necesitamos construir las funciolo son. De igual forma los marcos teóricos referenciales nes de utilidad individuales para cada stakeholder. Por que soportan los indicadores de competitividad van desejemplo, para el caso del sector de las Pymes en la prode la Teorı́a de Recursos y Capacidades [49], [35], hasta vincia de Pichincha, existen ocho sectores industriales los modelos de calidad [10]; [19]. de Pymes a considerar; sectores: textil, metalmecánico, En vista de que el peso especiﬁco de cada stakeholder graﬁco, alimenticio, quı́mico, de la construcción, madeno es el mismo, ya que no todos tienen los mismos dere- rero y de las Tic’s [12], y cada sector posee stakeholders chos ni son afectados de la misma manera por la gestión con caracterı́sticas especı́ﬁcas y distintivas, por lo que se de la empresa [56], se decidió diseñar un sistema de in- podrı́a diseñar un indicador particular para cada caso. dicadores que midan la competitividad de las Pymes, en La metodologı́a se iniciará determinando en el sector las la nueva perspectiva planteada aquı́, a través de varia- funciones de utilidad de cada uno de sus stakeholders. bles valoradas en una escala tipo Likert, utilizando en- Estas funciones de utilidad reﬂejan el conocimiento del cuestas personales dirigidas a los gerentes de las Pymes. experto o del consenso del grupo de expertos que conoEl indicador general considera a su vez varios compo- ce el sector a fondo, de todas formas, se necesita invesnentes o stakeholders: accionistas, clientes, empleados, tigación posterior para examinar aquellos factores que proveedores, acreedores, gobiernos y sociedad y medio inﬂuencian a un stakeholder a revelar su función de utiambiente. El indicador se desglosa en: variables, dimen- lidad a una ﬁrma [27]. siones e ı́tems. La medición se propone realizar con un Establecidas las funciones de utilidad para cada uno cuestionario de preguntas con respuestas de escala no- de los stakeholders, se procederá a determinar los pesos minal y de intervalo. Los puntajes máximos asignables a p en la función de preferencia, por el método de indifei los diferentes stakeholders se determinarán de acuerdo rencias. a la teorı́a de la decisión con múltiples objetivos [64].	19,566
hal-02280986-Kazanski_Umbo_Dobrodzien_2019.txt_1	French-Science-Pile	Open Science	Various open science	2,019	Умбоны типа Добродзень эпохи переселения народов: распространение, происхождение и датировка. Stratum plus. Archaeology and Cultural Anthropology, 2019, Меч Марса, 4, pp.155-167. ⟨hal-02280986⟩	None	Russian	Spoken	6,589	18,735	Умбоны типа Добродзень эпохи переселения народов: распространение, происхождение и датировка Michel Kazanski To cite this version: Michel Kazanski. Умбоны типа Добродзень эпохи переселения народов: распространение, происхождение и датировка. Stratum plus. Archaeology and Cultural Anthropology, 2019, Меч Марса, 4, pp.155-167. ⟨hal-02280986⟩ HAL Id: hal-02280986 https://hal.science/hal-02280986 Submitted on 6 Sep 2019 HAL is a multi-disciplinary open access archive L’archive ouverte pluridisciplinaire HAL, est desfor the deposit and dissemination of scientific re- tinée au dépôt et à la diffusion de documents sciensearch documents, whether they are published or not. tifiques de niveau recherche, publiés ou non, émanant The documents may come from teaching and research des établissements d’enseignement et de recherche institutions in France or abroad, or from public or pri- français ou étrangers, des laboratoires publics ou vate research centers. privés. Copyright E-ISSN: 1857-3533 №4. 2019 МЕЧ МАРСА Латная конница хунну и сяньби К истории Второй Боспорской войны Римский меч из Усть-Альминского некрополя Боевые ножи позднесарматского времени Умбоны эпохи переселения народов Воинские погребения могильника Чатыр-Даг Травмы как свидетельство вооруженного насилия Ответственный редактор — Олег В. Шаров САНКТ-ПЕТЕРБУРГ КИШИНЕВ ОДЕССА БУХАРЕСТ 2019 Stratum plus 155 №4. 2019 М. М. Казанский Умбоны типа Добродзень эпохи переселения народов: распространение, происхождение и датировка Keywords: Central Europe, Middle Danube, the Great Migration Period, shield boss, military burials, Stilicho Cuvinte cheie: Europa Centrală, Dunărea de Mijloc, epoca Marii migraţii a popoarelor, umboane, morminte de oșteni, Stilicon Ключевые слова: Центральная Европа, Средний Дунай, эпоха Великого переселения народов, умбоны, воинские погребения, Стилихон M. M. Kazanski The Shield Boss of Dobrodzień Type of the Great Migration Period: Distribution, Origin and Dating Shield bosses of Dobrodzień type are well known in Barbaricum, from the Danube to the eastern shore of the Black Sea. Most of these finds are from the barbaric sites of Central Europe, which are related to the Germans of the late Roman period and the beginning of the Great Migration period. Majority of Dobrodzien type shield bosses, regardless of the region they are found, come from “military” burials containing other weapons, in particular swords and spurs, both traditionally considered to be a sign of some military prestige. The time of their distribution corresponds to the initial phase of the Great Migration period, approximately 370—430. Even though all known shield boss of Dobrodzień type originate from a barbarian, primarily Germanic context, it is assumed that this type of weapon was of Roman origin, possibly representing production of Roman workshops on the Middle Danube. M. M. Kazanski Umboanele de tip Dobrodzień din epoca migraţiei popoarelor: răspândirea, provenienţa şi datarea Umboanele de tip Dobrodzień sunt bine cunoscute în Barbaricum, de la Dunăre şi până la ţărmul de răsărit al Mării Negre. Majoritatea descoperirilor acestor umboane provin din siturile barbare ale Europei centrale, puse în legătură cu germanicii din perioada romană târzie şi începutul epocii migraţiei popoarelor. Grosul umboanelor de tip Dobrodzień, indiferent de regiunea în care au fost descoperite, provin din morminte de oşteni, ce conţin şi alte piese de armament, în particular, spade şi pinteni, care tradiţional sunt considerate indiciu al prestigiului militar. Perioada lor de răspândire corespunde fazei incipiente a epocii migraţiei popoarelor, aproximativ anii 370—430. În pofida faptului că toate umboanele cunoscute provin din context barbar, în primul rând germanic, este presupusă provenienţa romană a acestui tip de armament, care reprezintă, posibil, o producţie a atelierelor romane de pe Dunărea de Mijloc. М. М. Казанский Умбоны типа Добродзень эпохи переселения народов: распространение, происхождение и датировка Умбоны типа Добродзень хорошо известны в Барбарикуме, от Дуная до восточного берега Черного моря. Большинство находок этих умбонов происходит с варварских памятников Центральной Европы, соотносимых с германцами позднеримского времени и начала эпохи переселения народов. Основная часть умбонов типа Добродзень, независимо от региона, где они обнаружены, происходит из «воинских» погребений, содержавших и другие предметы вооружения, в частности, мечи и шпоры, которые традиционно считаются признаком некоего воинского престижа. Время их распространения соответствует начальной фазе эпохи переселения народов, приблизительно 370—430 гг. Несмотря на то, что все известные умбоны происходят из варварского, в первую очередь германского контекста, предполагается римское происхождение этого типа оружия, возможно, представляющего собой продукцию римских мастерских на Среднем Дунае. Умбоны типа Добродзень (Dobrodzień) — Цилинг Т (Щукин 2011; Zieling 1989: 160—162) выделяются на фоне других тем, что имеют сфероконическую, довольно высокую калотту с вертикальными каннелюрами или фасетками и относительно неширокие поля. Здесь будет рассмотрен вопрос их возможного происхождения и уточнена датировка этого типа оружия. © Stratum plus. Археология и культурная антропология. © М. М. Казанский, 2019. Распространение Большинство находок этих умбонов происходит с варварских памятников Центральной Европы (рис. 1: 1), соотносимых с германцами позднеримского времени и начала эпохи переселения народов (Tejral 1986: Karte 1; Kazanski 1988: Annexe 2; Kokowski 1996: ryc. 2). Поэтому их иногда называют германскими (Feugère 156 М. М. Казанский Stratum plus №4. 2019 Рис. 1. Каpта распространения умбонов типа Добродзень (a) и география размещения scutaria на Дунае (b). 1 — Крикштонис; 2 — Добродзень; 3 — Ольштын; 4 — Мушов; 5 — Тисавалк; 6 — Тисакарад-Инаса; 7 — ТисадобСигед; 8 — Уйхартьян; 9 — Будешть; 10 — Залиски; 11 — Шапка-Абгыдзраху; 12 — Шапка-Ахьяцрарху; 13 — Марцианополь; 14 — Лауриакум; 15 — Карнунтум; 16 — Аквинкум; 17 — Сирмиум; 18 — Горреум Марги (a — по Istvánovits, Kulcsár 1992: kép. 17, 23; Kokowski 1996: ryc. 2; b — по Bishop, Coulston 1993: fig. 135). Fig. 1. Mapping of the Dobrodzień shield bosses (a) and distribution of scutaria on the Danube (b). 1 — Krikštonys; 2 — Dobrodzień; 3 — Olsztyn; 4 — Mušov; 5 — Tiszavalk; 6 — Tiszakarád-Inasa; 7 — Tiszadob-Sziged; 8 — Ujhartyán; 9 — Budeşti; 10 — Zaliski; 11 — Shapka-Abgydzrakhu; 12 — Shapka-Ahyatsarahu; 13 — Marcianopolis; 14 — Lauriacum; 15 — Carnuntum; 16 — Aquincum; 17 — Sirmium; 18 — Horreum Margi (a — after Istvánovits, Kulcsár 1992: kép. 17, 23; Kokowski 1996: ryc. 2; b — after Bishop, Coulston 1993: fig. 135). 1992: 248; Щукин 2011: 176). Впрочем, один умбон найден по соседству, в балтском контексте, в погребении на неманском могильнике Крикштонис (Приложение, №1) (рис. 2: 2). Фрагменты умбонов типа Добродзень к северу от Карпат найдены на могильниках т. н. добродзеньского типа — Добродзень и Ольштын (рис. 2: 6—9) в бассейне Верхней Вислы, в слое трупосожжений, связанных с заключительной фазой пшеворской культуры (Приложение, №2, 3). К югу от Карпат в первую очередь стоит назвать три умбона, обнаруженные в постройке на поселении Мушов в Южной Моравии (Приложение, №4) (рис. 2: 10—12). Также к югу от Карпат, в бассейне Верхней Тисы мне известно три находки таких умбонов, в погребениях с оружием на могильниках Тисавалк, Тисакарад-Инаса и Тисадоб-Сигед (рис. 3: 9; 4: 3, 10) (Приложение, №5—7). Одна находка, также в погребении с оружием, сопровождавшемся конским захоронением, отмечена несколько западнее, в Уйхартьян (рис. 4: 12), в междуречье Тисы и Дуная (Приложение, №8). Также в Карпатском бассейне, в Трансильвании, один умбон был обнаружен в Будешть (рис. 3: 11). Он, видимо, происходит из некрополя, где есть и другие предметы вооружения (Приложение, №9). В европейском Барбарикуме к востоку от Карпат известен лишь умбон, найденный на поселении римского времени Залиски (рис. 2: 5) около Львова (Приложение, №10). Наконец, сразу три умбона происходят с территории Абхазии. Они были найдены в Цебельдинской долине, в некрополе Шап- Stratum plus Умбоны типа Добродзень эпохи переселения народов 157 №4. 2019 Рис. 2. Находки в Крикштонис, погр. 2 (1—4), Залиски (5), Добродзень (6—8), Ольштын (9), Мушов, постр. 10 (10—12) (1—4 — по Kulikauskas 1959: Pav. 9; 5 — по Kokowski 1996: Ryc. 1; 6—8 — по Szydłowski 1974: Tabl. XCIII; XCV; 5 — по Szydłowski 1974: Tabl. CLXVII: f; 10—12 — по Trňáčková 1985: Abb. 1). Fig. 2. Finds in Krikštonys, grave 2 (1—4), Zaliski (5), Dobrodzień (6—8), Olsztyn (9), Mušov, build. 10 (10—12) (1—4 — after Kulikauskas 1959: Pav. 9; 5 — after Kokowski 1996: Ryc. 1; 6—8 — after Szydłowski 1974: Tabl. XCIII; XCV; 5 — after Szydłowski 1974: Tabl. CLXVII: f; 10—12 — after Trňáčková 1985: Abb. 1). ка, на кладбищах Абгыдзраху и Ахьяцараху (рис. 5: 1, 3; 6: 11) (Приложение, №11, 12). Для Понтийского региона стоит отметить и наличие других умбонов с фигурной калоттой. Так, умбоны с фасетками (тип Керчь) на калотте известны в Абхазии и в Керчи (Kazanski 1994: 448, An. 2: 18d, 21c). В Абхазии также были найдены два умбона с горизонтальными каннелюрами (тип Шапка) (Kazanski 1994: 448, An. 2: 21b, d). Возможно, 158 М. М. Казанский Stratum plus №4. 2019 Рис. 3. Находки в Тисадоб-Сигед, погр. 34 (по Istvánovits 1993: Abb. 14—16). Fig. 3. Finds from Tiszadob-Sziged, grave 34 (after Istvánovits 1993: Abb. 14—16). вариант умбона типа Добродзень (или параллельный тип?) был обнаружен в «княжеском» погребении Красная Поляна около Сочи, сопровождавшемся, в частности, серебряной сасанидской посудой. Однако каннелюры на умбоне из Красной Поляны не вертикальные, а наклонные (рис. 6: 9) (Kazanski 1994: 447, An. 2: 26). Легко заметить, что основная часть находок умбонов типа Добродзень, независимо от региона, где они обнаружены, происходит из «воинских» погребений (При- ложение, № 1, 5—8, 11, 12), содержавших и другие предметы вооружения, в частности, мечи и шпоры, и те и другие традиционно считаются признаком некоего воинского престижа. Говоря об умбонах типа Добродзень, необходимо упомянуть и их изображения. Наиболее известен диптих из Монцы (рис. 7: 1), который чаще всего связывают с известным римским военачальником вандальского происхождения Стилихоном и датируют его временем около 395 г. (подробнее см. Killerich, Stratum plus Умбоны типа Добродзень эпохи переселения народов 159 №4. 2019 Рис. 4. Находки в Тисавалк, погр. 6 (1—9), Тисакарад-Инаса (10), Будешть (11), Уйхартьян (12—15) (1—9 — по Garam, Vaday 1990: Abb. 10, 11; 10, 11 — по Istvánovits, Kulcsár 1992: Tábl. II: 1, II: 6; 12—15 — по Bóna 1961: Kép. 1). Fig. 4. Finds from Tiszavalk, grave 6 (1—9), Tiszakarád-Inasa (10), Budeşti (11), Ujhartyán (12—15) (1—9 — after Garam, Vaday 1990: Abb. 10, 11; 10, 11 — after Istvánovits, Kulcsár 1992: Tábl. II: 1; II: 6; 12—15 — after Bóna 1961: Kép. 1). Torp 1989, там же библиография). Хотелось бы еще раз (см. Казанский, Зальцман, Скворцов 2018: 16, рис. 18: 2) привлечь внимание исследователей и к отчетливому изображению умбона типа Добродзень на круглых накладках конского убора из всем известного погре- бения Унтерзибенбрунн (Untersiebenbrunn) в Нижней Австрии (рис. 7: 2) (см., напр. Tejral 2011: Abb. 125). Показательно, что на миниатюрном «умбоне» из Унтерзибенбрунна изображено такое же поле в виде звезды, как и на умбоне с диптиха из Монцы. Наход- 160 М. М. Казанский Stratum plus №4. 2019 Рис. 5. Находки в Шапка-Абгыдзраху, погр. 6 (2—13) и вне контекста (1) (по Трапш 1971: табл. III; XXXVI: 8). Fig. 5. Finds from Shapka-Abgydzrakhu, grave 6 (2—13) and out of context (1) (after Трапш 1971: табл. III; XXXVI: 8). ка из Унтерзибенбрунна является опорной для периода D2 по хронологии Барбарикума (380/400—400/450 гг.). Датировка На основании сравнительного анализа археологического контекста умбоны типа Добродзень отнесены к начальной фазе эпохи переселения народов, периоду D1 (360/370—400/410 гг.), точнее, ко времени око- ло 400 г. (Tejral 1986: 200, 203, 210; 2011: 123), что в целом соответствует эпохе Стилихона (убит 22 августа 408 г.). Согласно К. Годловскому, эти умбоны могут иметь несколько более широкую дату — они принадлежат группе 8 погребений пшеворской культуры с оружием и группе D погребений балтского круга, то есть относятся к концу периода С3 (хронологические рамки периода: 300/320—350/370 гг.) и началу периода D1 (Godłowski 1994: 170, 171, Abb. 1: 70; 2: 19). Stratum plus Умбоны типа Добродзень эпохи переселения народов 161 №4. 2019 Рис. 6. Находки в Красной Поляне (1—10) и Шапка-Ахьяцараху, погр. 20 (11—17) (1—10 — по Kazanski 1994: fig. 18: 1—9; 12—17 — по Трапш 1975: табл. XVI: 5; XIX: 1; XXII: 3; XXIII: 5; XXIV: 2, 12; 11 — по Воронов, Шенкао 1982: рис. 5—11). Fig. 6. Finds from Krasnaya Polyana (1—10) and Shapka-Ahyatsarahu, grave 20 (11—17) (1—10 — after Kazanski 1994: fig. 18: 1—9; 12—17 — after Трапш 1975: табл. XVI: 5; XIX: 1; XXII: 3; XXIII: 5; XXIV: 2, 12; 11 — after Воронов, Шенкао 1982: рис. 5—11). В целом к поздней части периода C3 и периоду D1 по совокупности находок могут быть отнесены «слойные» трупосожжения из Добродзень и Ольштына (Приложение, № 2, 3). Погребение 6 в могильнике Тисавалк, по сочетанию в инвентаре подвязной фибулы типа Альмгрен 162/Амброз 16-2-I-3 (Амброз 1966: 64—66) и пряжки с довольно коротким язычком (рис. 4: 6, 7), скорее датируется периодом С3 (Приложение, № 5). Абхазские погребения ШапкаАбгыдзраху 6 и Шапка-Ахьяцараху 20 (Приложение, № 11, 12) содержат дуговидные фибулы (рис. 5: 7; 6: 15), более типичные для стадий I/2 (260/270—330/340 гг.) и II/3—4 (320/330—400/410 гг.) местной абхазской хронологии (подробнее см. Kazanski, Mastykova 2007: 21, 22, 33, 34, Pl. 21: 25; 22: 162 М. М. Казанский Stratum plus №4. 2019 Рис. 7. Изображение на диптихе из Монцы (1) и накладки конской сбруи из Унтерзибенбрунна (2) (1 — по Killerich, Torp 1989: fig. 1: 15; 2 — по Tejral 2011: Abb. 125). Fig. 7. Image on diptych from Monza (1) and of horse harness from Untersibenbrunn (2) (1 — after Killerich, Torp 1989: fig. 1: 15; 2 — after Tejral 2011: Abb. 125). 8, 9), что соответствует периодам С3 и D1 1. Эти находки в целом подтверждают «широкую» датировку К. Годловского. 1 Иногда неправомерно утверждается, что хронологическая система, используемая здесь (периоды С и D), вырабатывалась только для центральноевропейского Барбарикума. Между тем, уже в публикации 1986 г. для разработки этой системы Ярославом Тейралом были привлечены находки c территории черняховской культуры, из Крыма, с Северного Кавказа и из Абхазии (Tejral 1986: 179—193, Abb. 1—7). В то же время погребение в могильнике Тисадоб (Приложение, №7) с умбоном типа Добродзень содержало двучастную подвязную фибулу с длинной расширенной ножкой и довольно короткой и широкой спинкой (рис. 3: 1). Она очень напоминает балканские подвязные фибулы позднего V — VI вв., типа ЮнцеВиминациум (Uenze 1992: 146—149, 151, Abb. 5: 1; 7: 1; Ivanišević, Kazanski, Mastykova 2006: 19; Kazanski, Mastykova 2017: 159), хотя и отличается от них более широким держателем пружины. Возможно речь идет об их про- Stratum plus Умбоны типа Добродзень эпохи переселения народов 163 №4. 2019 Рис. 8. Римские умбоны из Дура-Европос (по James 2004: fig. 95, N° 603, 604). Fig. 8. Roman shield bosses from Dura-Europos (after James 2004: fig. 95, no. 603, 604). тотипе, и в этом случае данная фибула вряд ли относится ко времени ранее V в. Подвязные фибулы с широкими спинкой и ножкой в Южной Cловакии известны в контексте периодов D1-D2 — это погр. 4 на могильнике Абрахам (Abrahám) (Tejral 2011: 103, Abb. 66). С другой стороны, фибула из Тисадоба напоминает и некоторые фибулы черняховской культуры — типа Амброз 16-2-II (Амброз 1966: 69), но у тех иные пропорции и иной способ крепления иглы. Точно так же фибулы с расширенными ножкой и спинкой и широким держателем иглы, известные в сарматских древностях позднеримского времени на территории Восточной Венгрии, имеют иные пропорции (сp. напр. Vaday 1989: Abb. 16: 9). Не исключено впрочем, что эти сарматские фибулы и лежат в начале эволюции балканских подвязных фибул. В Мушове (Приложение, №4) умбоны типа Добродзень были найдены в постройке, вместе с металлическим котлом, типичным для эпохи переселения народов (Tejral 1986: 220, Abb. 15: 3, 4; 2011: 123). Наконец, уже упоминавшееся миниатюрное изображение умбона типа Добродзень из Унтерзибенбрунна (рис. 7: 2), вне всякого сомнения, относится к периоду D2. Итак, по совокупности данных, умбоны типа Добродзень могут быть датированы в диапазоне от периода С3, скорее всего, его поздней части, по период D2. На мой взгляд, это соответствует приблизительно 370—430 гг. Вопрос происхождения Несмотря на то, что все известные умбоны происходят из варварского, в первую очередь германского контекста, мне представляется более обоснованным римское происхождение данного типа оружия, возможно, изначально связанное с мастерскими Паннонии и Норика. Аргументом в пользу римского происхождения умбонов является диптих из Монцы (рис. 7: 1). Мне кажется абсолютно невероятным, чтобы Стилихон, да и любой другой высокопоставленный варвар в Империи, которому мог принадлежать диптих, согласился бы на своем официальном портрете изобразить что-либо варварское. Всё, что мы знаем об интеграции варваров в римскую структуру власти, противоречит такой идее. «Варварская» материальная культура была совершенно неприемлема для западноримской властной элиты. В 417 г. был издан специальный декрет о запрещении ношения варварских причесок и варварской одежды из шкур/меха животных в Риме и соседних регионах (Code théodosien XIV, 10, 4, цит. по: Chastagnol 1976: 94), а император Грациан (375—383 гг.), по сообщению Аврелия Виктора (Epit. 47, 6), снискал ненависть своих солдат тем, что публично появлялся в аланской, т. е. варварской одежде (цит. по: Алемань 2003: 79, 80). Вряд ли такое сумасбродство мог себе позволить какой-либо генерал варварского происхождения на рим- 164 М. М. Казанский Stratum plus №4. 2019 ской службе. Следует напомнить, что Стилихона как раз и обвиняли в варварских симпатиях (Stein 1959: 253; Rutilus Namatianus, Sur le retour, II, 16—60, цит. по: Chastagnol 1976: 83, 84), что в конечном итоге и послужило поводом к его убийству. Иногда говорят, что присутствие «варварского» умбона на диптихе из Монцы свидетельствует о распространении новой моды, запущенной предводителями варваров-федератов (Каргопольцев, Бажан 1993: 117). Такая мода в эпоху переселения народов действительно была, о ней на территории Западной Римской империи свидетельствуют погребения воинских предводителей горизонта Унтерзибенбрунн, такие как Мундольсхейм (Mundolsheim), Фюрст (Fürst), Лебень (Lébény-Magasmart), Ленделтоть (Lengyeltóti) (подробнее см. Казанский 2010). Однако представленные этими погребениями вожди варваров-федератов до ранга Стилихона, да и вообще римского генералитета, явно «не дотягивали» 2. Они, по сути, оставались со своими варварами вне рамок римского общества и официальных диптихов себе не заказывали. Если же конкретно говорить о Стилихоне, то он никакого отношения к предводителям федератов не имел, а был главнокомандующим регулярных войск Западной Римской империи (Stein 1959: 226). Следует отметить, что характерные поля умбона в виде звезды, изображенные на диптихе из Монцы, имеют параллели на достоверно римских умбонах первой половины III в. н. э. из известной крепости Дура-Эвропос на Евфрате (рис. 8) (James 2004: 175, fig. 95, N° 603, 604; см. также: Przybyła 2014: 217) и не известны на умбонах из Барбарикума. Дунайские провинции Империи, и в первую очередь Паннония и Норик, как место производства умбонов типа Добродзень, а вернее, их прототипов, подходят с точки зрения географии, поскольку они находятся как бы в эпицентре распространения умбонов (рис. 1). Напомним, что именно в этих дунайских провинциях Империи, согласно Noticia Dignitatum, в частности, Лауриакуме, Карнунтуме, Аквинкуме, Сирмиуме находилась основня часть государственных мастерских, scutaria, по производству щитов для римской армии (рис. 1: 2). Кроме того, есть и другие элементы материальной культуры Барбарикума, явно распространявшиеся из дунайских римских провинций, как, например, вещи со штампованным декором (подробнее см. Казанский, Мастыкова 2018, там же библиография). Еще один аргумент в пользу римской, а не варварской атрибуции умбонов типа Добродзень — их появление на восточном берегу Черного моря (рис. 1: 11, 12). Германские элементы в эпоху переселения народов в этом регионе представлены, но, во-первых, их там меньше, чем принято считать, а во вторых, эти германские элементы появились на восточном берегу Черного моря вместе с римской армией (подробнее см. Казанский 2015). Приложение. Находки умбонов типа Добродзень 1. Крикштонис (Krikštonys), погр. 2. Kulikauskas 1959: Pav. 9; Казакявичюс 1988: 126, 128; Zieling 1989: №1961. 2. Добродзень (Dobrodzień-Rędzina), могильник, слой трупосожжений. Szydłowski 1974: 77, Tabl. XCIII, XCV; Zieling 1989: №996—1001. 3. Ольштын (Olsztyn), могильник, слой трупосожжений. Szydłowski 1974: 141, Tabl. CLXVII: е, f; Zieling 1989: №1319, 1320. 4. Мушов (Mušov), постройка 10. 2 В частных беседах известный историк Барбарикума Хервиг Вольфрам характеризовал воинских предводителей, погребенных в этих могилах, как «полковников», а выдающийся венгерский археолог, ныне покойный Аттила Кишш, квалифицировал их как «председателей варварских колхозов». Trňáčková 1985; Tejral 1986: Abb. 15: 1, 2; Tejral 2011: Abb. 79: 1, 3, 4; 297. 5. Тисавалк (Tiszavalk), погр. 6. Garam, Vaday 1990: Abb. 10, 11; Istvánovits, Kulcsár 1992: 63, Tábl. II: 3. 6. Тисакарад-Инаса (Tiszakarád-Inasa), погр. 35. Istvánovits, Kulcsár 1992: 63, Tábl. II: 1. 7. Тисадоб-Сигед (Tiszadob-Sziged), погр. 34. 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La nécropole apsile de Tsibilium (VIIe av. J.-C. — VIIe ap. J.-C.) (Abkhazie, Caucase). Vol. 2. L’étude du site. BAR IS S1721-2. Oxford: John and Erica Hedges Ltd. Kazanski M., Mastykova A. 2017. La culture matérielle. VI. Objets en métal. In: Popović I., Kazanski M., Ivanišević V. (dir.). Sirmium à l’époque des Grandes Migrations. Centre de recherche d’histoire et civilisation de Byzance. Monographies 53. Leuven; Paris; Bristol: Peeters, 157—182. Killerich B., Torp H. 1989. Hic est: hic Stilicho. The Date and Interpretation of a Notable Diptych. Jahrbuch des Deutschen Archäologischen Instituts 104, 319—371. Kokowski A. 1996. W kwestii rozprzestreniania się umb facetowana pokrywa (typ Dobrodzień). Studia i materiały do dziejów Kujaw — Niźu Polski 6, 179—187. Kulikauskas P. 1959. Naujas archeologinis pamiklas Užneminėje (V—VII amžų jotvingių senkapis Krikštonyse, Ladzijų raj.). Lietuvos TSR Moksų akademijos darbai, A serija 1, 71—88. Przybyła M. J. 2014. Foederati na Zelandii? 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Das norddanubi- sche Gebiet zur Zeit der Völkerwanderung. Brno: Archäologisches Institut AW CR. Trňáčková Z. 1985. Ein Hortfund von metallgegenständen aus der späten Kaiserzeit und frühen Völkerwanderungszeit aus Mušov. Památky archeologické LXXXVI, 279—284. Uenze S. 1992. Die spätantiken Befestigungen von Sadovec (Bulgarien). München: C. H. Beck’sche Verlagbuchhandlung. Vaday A. 1989. Die sarmatischen Denkmäler des Komitats Szolnok. Ein Beitrag zur Archäologie und Geschichte des sarmatischen Barbaricums. Antaeus 17—18. Budapest: Archäologisches Institut der UAW. Zieling N. 1989. Studien zu germanischen Schilden der Spätlatène, und der römischen Kaiserzeit im freien Germanien. BAR IS 505. Oxford: BAR. References Alemany, A. 2003. Alany v drevnikh i srednevekovykh pis’mennykh istochnikakh (Sources on the Alans: A Critical Compilation). Moscow: “Menedzher” Publ. (in Russian). Ambroz, A. K. 1966. Fibuly iuga evropeiskoi chasti SSSR (II v. do n. e. — IV v. n. e.) (Fibulae of the South of European Part of USSR (2nd Century BC — 4th Century AD)). Series: Svod Arkheologicheskikh Istochnikov (Corpus of Archaeological Sources) D1—30. Moscow: “Nauka” Publ. (in Russian). Voronov, Yu. N., Shenkao, N. K. In Ambroz, A. K., Erdely, I. (eds.). 1982. Drevnosti epokhi velikogo pereseleniia narodov V—VIII vv. Sovetsko-vengerskii sbornik (Antiquities of the Great Migrations in 5th —8th Centuries. Soviet-Hungarian Volume). Moscow: “Nauka” Publ., 121—165 (in Russian). Kazakevičius, V. 1988. Oruzhie baltskikh plemen II—VIII vekov na territorii Litvy (Weapons of the Baltic Tribes in 2nd —8th Centuries on the Territory of Lithuania). Vilnius: “Mokslas” Publ. (in Russian). Kazanski, M. M. 2010. In Nosov, E. N., Beletsky, S. V. (manag. eds.). Kraeugol’nyi kamen’. Arkheologiia, istoriia, iskusstvo, kul’tura Rossii i sopredel’nykh stran (Corner-Stone. Archaeology, History, Arts and Culture of Russia and Its Neighbours) I. Moscow: “Lomonosov” Publ., 307—320 (in Russian). Kazanski, M. M. 2015. In Scripta Antiqua 4, 33—60 (in Russian). Kazanski, M. M., Zal’tsman, E. B., Skvortsov, K. N. 2018. Rannesrednevekovyi mogil’nik Zaostrov’e-1 v Severnoi Sambii (Zaostrovye-1, the Early Medieval Burial Ground in Northern Sambia). Moscow: Institute of Archaeology, Russian Academy of Sciences (in Russian). Kazanski, M. M., Mastykova, A. V. 2018. In Bosporskie issledovaniia (Bosporan Studies) XXXVI, 118—142 (in Russian). Kargopoltsev, S. Yu., Bazhan, I.A. 1993. In Peterburgskii arkheologicheskii vestnik (St. Petersburg Archaeological Bulletin) 2, 113—126 (in Russian). Trapsh, M. M. 1971. In Trapsh, M. M. Trudy (Works) III. Tbilisi: “Metsniereba” Publ. (in Russian). Trapsh, M. M., 1975. In Trapsh, M. M. Trudy (Works) IV. Sukhumi: “Metsniereba” Publ., 9—87 (in Russian). Shchukin, M. B. 2011. In Stratum plus. Archaeology and Cultural Anthropology (4), 167—178 (in Russian). Bishop, M. C., Coulston, J. C. N. 1993. 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Das Gräberfeld aus dem 4—5. Jahrhundert von Tiszadob-Sziget. Acta Archaeologica Academiae Scientiarum Hungaricae 45, 42—146. Istvánovits, E., Kulcsár, V. 1992. Pajzsos temetkezések a Dunától keletre eső kárpát-medencei Barbaricumban. A Nyíregyházi Jósa András Múzeum évkönyve XXX—XXXII, 47—96. Ivanišević, V., Kazanski, M., Mastykova, A. 2006. Les nécropoles de Viminacium à l’époque des Grandes Migrations. Monographies du Centre de Recherche d’Histoire et Civilisation de Byzance — Collège de France 22. Paris: Centre de Recherche d’Histoire et Civilisation de Byzance. James, S. 2004. The Excavations at Dura-Europos conducted by Yale University and the French Academy of Inscriptions and Lettres, 1928 to 1937. Final Report VII. The Arms and Armour and other Military Equipment. London: British Museum Press. Kazanski, M. 1988. Quelques parallèles entre l’armement en Occident et à Byzance (IVe — VIIe s.). In Landes, C. (dir.). Gaule mérovingienne et monde méditerranéen. Les derniers Romains en Septimanie, IVe — VIIIe siècles. Lattes: Imago, 75—87. Kazanski, M. 1994. Les éperons, les umbo, les manipules de boucliers et les haches de l’époque romaine tardive dans la région pontique: origine et diffusion. In Carnap-Bornheim, C. v. (Hrsg.). Beiträge zur römischer und barbarischer Bewaffnung in der ersten vier nachchristlichen Jahrhunderten. Lublin; Marburg: Vorgeschichtliches Seminar der PhillipsUniversität Marburg, 429—485. Kazanski, M., Mastykova, A. 2007.’La nécropole apsile de Tsibilium (VIIe av. J.-C. — VIIe ap. J.-C.) (Abkhazie, Caucase). Vol. 2. L’étude du site. BAR IS S1721—2. Oxford: John and Erica Hedges Ltd. Kazanski, M., Mastykova, A. 2017. La culture matérielle. VI. Objets en métal. In Popović, I., Kazanski, M., Ivanišević, V. (dir.). Sirmium à l’époque des Grandes Migrations. Centre de recherche d’histoire et civilisation de Byzance. Monographies 53. Leuven; Paris; Bristol: Peeters, 157—182. Killerich, B., Torp, H. 1989. Hic est: hic Stilicho. The Date and Interpretation of a Notable Diptych. Jahrbuch des Deutschen Archäologischen Instituts 104, 319—371. Kokowski, A. 1996. W kwestii rozprzestreniania się umb facetowana pokrywa (typ Dobrodzień). Studia i materiały do dziejów Kujaw — Niźu Polski 6, 179—187. Stratum plus Умбоны типа Добродзень эпохи переселения народов 167 №4. 2019 Kulikauskas, P. 1959. Naujas archeologinis pamiklas Užneminėje (V—VII amžų jotvingių senkapis Krikštonyse, Ladzijų raj.). Lietuvos TSR Moksų akademijos darbai, A serija 1, 71—88. Przybyła, M. J. 2014. Foederati na Zelandii? Analiza uzbrojenia pochówku z pierwszej poł. III w. po Chr. z Aasø, Zelandia. In Madyda-Legutko, R., Rodzińska-Nowak, J. (eds.). Honoratissimum assensus genus est armis laudare Studia dedykowane Profesorowi Piotrowi Kaczanowskiemu z okazji siedemdziesiątej rocznicy urodzin. Kraków: Uniwersytet Jagielloński, Instytut Archeologii, 213—224. Stein, E. 1959. Histoire du Bas-Empire. Tome premier. De l’Etat Romain à l’Etat Byzantin (284—476). Paris: Desclée de Brouwer. Szydłowski, J. 1974. Trzy cmentarzyska typu dobrodzieńskiego. Rocznik Muzeum górnośląskiego w Bytomiu 11). Bytom: Muzeum górnośląskie w Butomiu. Tejral, J. 1986. Fremde Einflüsse und kulturelle Veränderungen nördlich der mittlerer Donau zu Beginn der Völkerwan- derungszeit. In Kmieciński, J. (Hrsg.). „Peregrinatio Gothica“. Archaeologia Baltica VII. Łódź: Katedra archeologii Uniwersytetu Łódzkiego, 175—238. Tejral, J. 2011. Einheimische und Fremde. Das norddanubische Gebiet zur Zeit der Völkerwanderung. Brno: Archäologisches Institut AW CR. Trňáčková, Z. 1985. Ein Hortfund von metallgegenständen aus der späten Kaiserzeit und frühen Völkerwanderungszeit aus Mušov. Památky archeologické LXXXVI, 279—284. Uenze, S. 1992. Die spätantiken Befestigungen von Sadovec (Bulgarien). München: C. H. Beck’sche Verlagbuchhandlung. Vaday, A. 1989. Die sarmatischen Denkmäler des Komitats Szolnok. Ein Beitrag zur Archäologie und Geschichte des sarmatischen Barbaricums. Antaeus 17—18. Budapest: Archäologisches Institut der UAW. Zieling, N. 1989. Studien zu germanischen Schilden der Spätlatène, und der römischen Kaiserzeit im freien Germanien. BAR IS 505. Oxford: BAR. Статья поступила в номер 1 марта 2019 г. Michel Kazanski (Paris, France). Doctor Habilitat on Archaeology. Centre national de la recherche scientifique, UMR 8167: Centre d’Histoire et Civilisation de Byzance 1. Mihail Kazanski (Paris, Franţa). Doctor habilitat în arheologie. Centrul naţional de cercetări ştiinţifice, Centrul de istorie şi civilizaţie al Bizanţului. Казанский Михаил Михайлович (Париж, Франция). Доктор хабилитат археологии. Национальный центр научных исследований, Центр истории и цивилизации Византии. E-mail: michel.kazanski53@gmail.com Address: 1 Rue du Cardinal Lemoine, 52, Paris, 75005, France Stratum plus 407 №4. 2019 Список сокращений АНИИ ЭЯЛИ АЕС АИАПМЗ АлтГУ АМЗ АН АН СССР АНО «НИЦ» АО АСГЭ АЭАЕ БелГУ БИ БКИЧП БФАН СССР ВАУ ВГПУ ВГУ ВДИ ВИМАИВиВС ВОКМ ВолГУ ВООПИиК ГАГУ ГАИМК ГАРО ГИМ ГИМЮУ ГМЗ ГРВЛ ГЭ ДГПБ ДГТУ ДГУ ЖМНП ЗООИД ИА АН РТ ИА АН УССР ИА КН МОН РК ИА НАНУ ИА РАН ИАИ ИАИАНД ИАК ИАН ИАЭТ СО РАН ИВИ РАН ИИКНП УдГУ ИИМК РАН — Адыгейский научно-исследовательский институт экономики, языка, литературы и истории. Майкоп. — Археология евразийских степей. Казань. — Азовский историко-археологический и палеонтологический музей-заповедник. Азов. — Алтайский государственный университет. Барнаул. — Азовский музей-заповедник. Азов. — Академия наук — Академия наук СССР. Москва. — Автономная некоммерческая организация «Научно-исследовательский центр по сохранению культурного наследия». Саратов. — Археологические открытия. 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Санкт-Петербург. — Музей антропологии и этнографии АН СССР / РАН им. Петра Великого (Кунсткамера). Ленинград / Санкт-Петербург. — Московский государственный университет им. М. В. Ломоносова. Москва. — Материалы и исследования по археологии СССР. Москва. — Материалы и исследования по археологии Кубани. Краснодар. — Материалы и исследования по археологии России. Москва. — Материалы и исследования по археологии юга России. Ростов-на-Дону. — Материалы по изучению историко-культурного наследия Северного Кавказа. Москва. — Международный институт центральноазиатских исследований ЮНЕСКО. Самарканд. — Нижневолжский археологический вестник. Волгоград. — Новосибирский государственный университет. Новосибирск. — Новочеркасский музей истории донского казачества. Новочеркасск. — Нумизматика и эпиграфика. Москва. — Объединение государственных книжно-журнальных издательств. — Одесское общество истории и древностей. Одесса. — Поволжская археология. Казань. — Петербургский археологический вестник. Санкт-Петербург. — Петербургский / Петроградский историко-филологический институт. Санкт-Петербург / Петроград. — Проблемы истории, филологии и культуры. Магнитогорск. — Российская археология. Москва. — Российский археологический ежегодник. Санкт-Петербург. — Российская академия истории материальной культуры. Ленинград. — Российская академия народного хозяйства и государственной службы при Президенте Российской Федерации. Москва. — Российский гуманитарный научный фонд. Москва. — Раннеславянский мир. Москва. — Советская археология. Москва. — Свод археологических источников. Москва; Ленинград. — Самбийская археологическая экспедиция ИА РАН. — Саратовский государственный университет. Саратов. — Сообщения Государственного Эрмитажа. Ленинград / Санкт-Петербург. — Самарский научный центр Российской Академии наук. Самара. — Самарский областной историко-краеведческий музей. Самара. — Санкт-Петербургский государственный университет. Санкт-Петербург. — Санкт-Петербургский филиал Архива Российской академии наук. Санкт-Петербург. — Тверской археологический сборник. Тверь. — Таманская археологическая экспедиция ИА РАН. — Труды Оренбургской учёной архивной комиссии. Оренбург. Stratum plus Список сокращений 409 №4. 2019 УАВ — Уфимский археологический вестник. Уфа. УдНИИЯЛ УрО РАН — Удмуртский научно-исследовательский институт истории, языка и литературы Уральского отделения Российской Академии наук. УрГУ — Уральский государственный университет. Свердловск / Екатеринбург. УрО РАН — Уральское отделение Российской Академии наук. Ижевск. ХС/ХСб — Херсонесский сборник. Севастополь. ЧелГУ — Челябинский государственный университет. Челябинск. ЧИ НИИЯЛИ — Чечено-Ингушский научно-исследовательский институт языка, литературы и истории. Грозный. ЭО — Этнографическое обозрение. Москва. ЮНЦ РАН — Южный научный центр Российской Академии наук. Ростов-на-Дону. ЮУрГУ — Южноуральский государственный университет. Челябинск. AV CR — Akademie věd České republiky. Praha. BAR — British Archaeological Reports. London; Oxford. BAR IS — British Archaeological Reports, International Series. London; Oxford. BayVgBl — Bayerische Vorgeschichtsblätter. München. BCH — Bulletin de Correspondance Hellénique. Paris; Athènes. CIRB — Corpus Inscriptionum Regni Bosporani = Корпус боспорских надписей. Москва; Ленинград. DFG — Deutsche Forschungsgemeinschaft. Bonn. ESA — Eurasia Septentrionalis Antiqua. Helsinki. IA UW — Instytut Archeologii Uniwersytetu Warszawskiego. Warszawa. IOSPE — Inscriptiones orae septentrionalis Ponti Euxini. Petropoli. JbGPV — Jahresbericht der Gesellschaft Pro Vindonissa. Brugg. LIMC — Lexicon Iconographicum Mythologiae Classicae. Zurich. MTA — Magyar Tudományos Akadémia. Budapest. PU SA IHMC RAS — Photo Unit of the Scientific Archive of the Institute for the History of Material Culture of the Russian Academy of Sciences. Saint Petersburg. RASMI — Rassegna di Studi del Civico Museo Archeologico e del Civico Cabinetto Numismatico di Milano. Milano. RGZM — Römisch-Germanisches Zentralmuseum Mainz. SAV — Slovenská Akadémia Vied. Bratislava. SC — Scythica et Caucasica e veteribus scriptoribus Graecis et Latinis collegit et cum versione Rossica edidit Basilius Latyschew. UAW — Ungarische Akademie der Wissenschaften. Budapest. UMCS — Uniwersytet Marii Curie-Skłodowskiej. Lublin. ZPE — Zeitschrift für Papyrologie und Epigraphik. Köln.	48,793
https://openalex.org/W4287364139	OpenAlex	Open Science	CC-By	2,021	A study on Time Sensitive Data Access Control	Dr Piyush Kumar Pareek	English	Spoken	1,600	2,981	INTRODUCTION INTRODUCTION Ciphertext policy attribute-based encryption (CP-ABE) which is a significant cryptographic technique for accessing the data and controlling the storage of data in the cloud. The cryptographic techniques will be utilized for providing the security for the information put present in the cloud. The CP-ABE technique permits the information get to control in the distributed storage [4]. The CP-ABE based strategies enables the information proprietors to empower the all-around characterized and stable control on the possess information display in the cloud. The primary issue emerges in the CP-ABE is to decide the clients get to in light of different factors, for example, username, date of birth and so forth., however here in the current framework there is no much spotlight on the time factor. In CP-ABE plot, everybody can get to information whenever in the event that they have a key to get to the private information in broad daylight cloud and it utilizes additional time and asset devouring. Continuously situations time factor assumes a basic part in time delicate information. Journal of Advance Research in Mobile Computing Volume 2 Issue 3 Journal of Advance Research in Mobile Computing Volume 2 Issue 3 Corresponding Author E-mail Id:-piyushkumarpareek88@gmail.com Corresponding Author E-mail Id:-piyushkumarpareek88@gmail.com A study on Time Sensitive Data Access Control Dr Piyush Kumar Pareek 1*, Chaitra Y R 2, Saumya L 3 1Professor, 2,3Assistant Professor CSE, East West Institute of Technology, Bengaluru, India. ABSTRACT To properly protect stored private data, the device proprietor will exchange private data with a chosen few and issue decipherment keys to them. If the user has quit, the cloud will be permitted to compensate them in re-scrambling the details and will even design new decipherment keys for current customers so that they will receive the information. Since a circulated registering environment is affecting all the different cloud servers, identical charges can't be got, and death by several of the cloud servers because of competing framework trades. The suggested approach would require learning, that will be re- scrambled, at various sections of time. The guarantee is rendered on high modern cryptography basis, with strong security emphasis, so that fine-grained knowledge gets to every business, and they don't have to get intense coordination for precision. Keywords:-decipherment keys, cloud servers, Ciphertext policy attribute-based encryption (CP-ABE), Decryption Keywords:-decipherment keys, cloud servers, Ciphertext policy attribute-based encryption (CP-ABE), Decryption REFERENCES 1. Wan, Z., & Deng, R. H. (2011). HASBE: A hierarchical attribute- based solution for flexible and scalable access control in cloud computing. IEEE transactions on information forensics and security, 7(2), 743-754. 1. Wan, Z., & Deng, R. H. (2011). HASBE: A hierarchical attribute- based solution for flexible and scalable access control in cloud computing. IEEE transactions on information forensics and security, 7(2), 743-754. Authority creates the token key to openly distribute a period for the respective token TKt as TKt = H1(t)γ. Authority creates the token key to openly distribute a period for the respective token TKt as TKt = H1(t)γ. different encryption types of comparable data, which puts repetitive and considerable load on the data proprietor. Data access security is more important for the data present in the cloud [7]. Here we are inheriting CP-ABE and designing an approach in which efficient access control policy is being emphasized with various access requirements focused on sensitive data. The scheme focuses on the trapdoor- based search mechanism where the data user search for the particular data present in the cloud according to the trapdoor the information will be shown but the user needs to decrypt the data which means the particular central authority will distribute the access to the user in particular time and in that time-slot only the user needs to decrypt the data and access it and as the time lapses, the data which is been uploaded by the data owner cannot be viewed [3]. 5. Trapdoor introduction: Basically, it is search operation performed on the database. A data user can perform this operation, where he will not be able to access the data without the permission of the owner of the data but he can view the file name, but he will not have access to open that file. 5. Trapdoor introduction: Basically, it is search operation performed on the database. A data user can perform this operation, where he will not be able to access the data without the permission of the owner of the data but he can view the file name, but he will not have access to open that file. Decryption (PK,CT,SK): After the data is encrypted, to decrypt the data which will be in cipher text format, we will require a decryption key. It depends on users to users, whether the user is a data owner or a data user. Proving a required decryption key is mandatory tp restore the data by following various form of properties. LITERATURE SURVEY To comprehend the utilization on time- based capacity, it is fundamental to display a capable arrangement, which won't release the data to get the opportunity to profit to expected customers until the point that accomplishing predefined time centers. A minor course of action is to allow the data proprietors physically discharge the sensitive form of data based on time. The proprietor exchanges the varied collection of data underneath, by using different techniques at each time with the ultimate objective that the normal customers can't get to the data until the point that the relating time arrives. Regardless, this plan controls the proprietor to on and on exchange the HBRP Publication Page 1-3 2020. All Rights Reserved Page 1 Journal of Advance Research in Mobile Computing Volume 2 Issue 3 Journal of Advance Research in Mobile Computing Volume 2 Issue 3 CONCLUSION Our proposed framework comprises of six parameters for encryption and decryption of the data: Our proposed framework comprises of six parameters for encryption and decryption of the data: 1. Setup: The setup calculation requires some serious energy as info. It creates the yield as people in general parameters PK and an ace key MK. 2. Yang K., Jia X., Ren X., Zhang B., Xie, R.(2013). DACMACS: Effective data access control for multi-authority cloud storage systems. Transactions on Information Forensics and Security,8(2),1790–1801. 2. Key Generation (MK, S): Various form of keys are generated in this phase, such as pub1ic key and secret key. The key time computation occurs as form of access key MK and an array of property S that depicts the key. It produces a private key SK. 3. Bertino, E., Bonatti, P. A., & Ferrari, E. (2001). TRBAC: A temporal role- based access control model. ACM Transactions on Information and System Security (TISSEC), 4(3), 191- 233.. 3. Encryption (PK, A, M): In this phase, a data will be encrypted using encryption algorithm and those are shown as a cipher text document and after the data is been encrypted, to decrypt it, a secret key is required. The encryption phenomenon usually occurs on the data which is pub1ic that is PK. 3. Encryption (PK, A, M): In this phase, a data will be encrypted using encryption algorithm and those are shown as a cipher text document and after the data is been encrypted, to decrypt it, a secret key is required. The encryption phenomenon usually occurs on the data which is pub1ic that is PK. 4. Li, M., Yu, S., Zheng, Y., Ren, K., & Lou, W. (2012). Scalable and secure sharing of personal health records in cloud computing using attribute-based encryption. IEEE transactions on parallel and distributed systems, 24(1), 131-143. 4. Token generation: Serially at each point of time that is t ∈ FT, Central 4. Token generation: Serially at each point of time that is t ∈ FT, Central HBRP Publication Page 1-3 2020. All Rights Reserved Page 2 Journal of Advance Research in Mobile Computing Volume 2 Issue 3 Journal of Advance Research in Mobile Computing Volume 2 Issue 3 10. Liu, Q., Wang, G., & Wu, J. (2014). Time-based proxy re-encryption scheme for secure data sharing in a cloud environment. Information sciences, 258, 355-370. 5. Zhou, Z., Zhang, H., Zhang, Q., Xu, Y., & Li, P. (2014, December). Privacy-preserving granular data retrieval indexes for outsourced cloud data. In 2014 IEEE Global Communications Conference (pp. 601-606). IEEE. 11. Xu, L., Zhang, F., & Tang, S. (2014). Timed‐release oblivious transfer. Security and Communication Networks, 7(7), 1138-1149. 6. Liu, Q., Tan, C. C., Wu, J., & Wang, G. (2011, December). Reliable re- encryption in unreliable clouds. In 2011 IEEE Global Telecommunications Conference- GLOBECOM 2011 (pp. 1-5). IEEE. 12. Androulaki, E., Soriente, C., Malisa, L., & Capkun, S. (2014, June). Enforcing location and time-based access control on cloud-stored data. In 2014 IEEE 34th International Conference on Distributed Computing Systems (pp. 637-648). IEEE. 7. Bethencourt, J., Sahai, A., & Waters, B. (2007, May). Ciphertext-policy attribute-based encryption. In 2007 IEEE symposium on security and privacy (SP'07) (pp. 321-334). IEEE. 13. Fan, C. I., & Huang, S. Y. (2014). Timed-release predicate encryption and its extensions in cloud computing. 15(3), 413-425. 8. Boneh, D., & Franklin, M. (2001, August). Identity-based encryption from the Weil pairing. In Annual international cryptology conference (pp. 213-229). Springer, Berlin, Heidelberg. 14. Zhu, X., Shi, S., Sun, J., & Jiang, S. (2014). Privacy-preserving attribute- based ring signcryption for health social network. In Proceedings of the 2014 IEEE Global Communications Conference (GLOBECOM2014) (pp. 3032-3036). 9. Yuan, K., Liu, Z., Jia, C., Yang, J., & Lv, S. (2013, September). Public key timed-release searchable encryption. In 2013 Fourth International Conference on Emerging Intelligent Data and Web Technologies (pp. 241- 248). IEEE. 15. https://hipaa.yale.edu/security/break- glass-proceduregranting-emergency- access-critical-ephi-systems HBRP Publication Page 1-3 2020. All Rights Reserved Page 3	19,596
W4255490701.txt_34	Open-Science-Pile	Open Science	Various open science	2,014	phase-space theory	None	English	Spoken	8,014	13,437	See also: cyanides, isocyanides, carbonitriles Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1350 nitrilium betaines Derivatives of nitriles having the general structure R–C≡N+–Y−. A subclass of 1,3-dipolar compounds including nitrile imides, nitrile oxides, nitrile sulfides and nitrile ylides. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitrilium ions Cations derived formally by attachment of one hydron to the nitrogen atom of a nitrile and hydrocarbyl derivatives thereof. E.g. benzonitrilium. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitrimines Compounds having the structure O2NN=CR2 (also called N-nitroimines). IUPAC Compendium of Chemical Terminology 995 of 1622 Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitro compounds Compounds having the nitro group, –NO2 (free valence on nitrogen), which may be attached to carbon, nitrogen (as in nitramines), or oxygen (as in nitrates), among other elements (in the absence of specification, C-nitro compounds are usually implied). See also: dipolar compounds Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitrogen fixation The assimilation of atmospheric nitrogen by microbial reduction to ammonia and conversion into organonitrogen compounds such as amino acids. Only a limited number of microorganisms are able to fix nitrogen. Source: PAC, 1992, 64, 143 (Glossary for chemists of terms used in biotechnology (IUPAC Recommendations 1992)) on page 161 nitrogen laser A source of pulsed semi-coherent superradiance mainly around molecular nitrogen. See: gas lasers . The lasing species is Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2256 nitrolic acids Compounds having the structure RC(=NOH)NO2. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitrones The N-oxides of imines, that have the structure R2C=N+(O−)R' (R' ≠ H). Synonymous with azomethine oxides. N-Oxides R2C=N+(O−)H may be included. IUPAC Compendium of Chemical Terminology 996 of 1622 See also: dipolar compounds Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1351 nitrosamides Although this term has, regrettably, been used to mean the N-nitroso derivatives of amides, logically it refers to amides of nitrous acid; the unstable parent compound H2NNO is, in fact, named nitrosamide, but its derivatives formed by substitution with hydrocarbyl groups are by long custom called nitrosamines. See also: nitramines Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nitrosamines N-Nitroso amines, compounds of the structure R2NNO. Compounds RNHNO are not ordinarily isolable, but they, too, are nitrosamines. The name is a contraction of N-nitrosoamine and, as such, does not require the N locant. See also: nitrosamides Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nitrosimines N-Nitroso imines O=NN=CR2. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nitroso compounds Compounds having the nitroso group, –NO. attached to carbon, or to another element, most commonly nitrogen or oxygen. See: nitrosamines Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 IUPAC Compendium of Chemical Terminology 997 of 1622 nitrosolic acids Compounds having the structure RC(=NOH)NO. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nitroxides Nitroxide is the parent name used by Chemical Abstracts Service for H2N–O., e.g. (ClCH2)2N–O. bis(chloromethyl) nitroxide. The IUPAC name is bis(chloromethyl)aminoxyl. Nitroxides should not be used as a class name for aminoxyl radicals. See: aminoxyl radicals Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nitroxyl radicals See: aminoxyl radicals Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 no carrier added A preparation of a radioactive isotope which is essentially free from stable isotopes of the element in question. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2522 no observed adverse effect level (NOAEL) Greatest concentration or amount found by experiment or observation, which causes no detectable adverse alteration of morphology, functional capacity, growth, development, or life span of the target organism under defined conditions. Source: PAC, 1993, 65, 2003 (Glossary for chemists of terms used in toxicology (IUPAC Recommendations 1993)) on page 2076 PAC, 2001, 73, 993 (Risk assessment for occupational exposure to chemicals. A review of current methodology (IUPAC Technical Report)) on page 1023 This definition replaces an earlier definition of no-observed-adverse-effect-level (NOAEL). IUPAC Compendium of Chemical Terminology 998 of 1622 no-bond resonance See: hyperconjugation Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1145 no-load indication (for a precision balance) The deflection or rest point (no-load reading) multiplied by the value of the division for zero load (rider at zero); symbol. Source: Orange Book, p. 36 no-observed-effect-level (NOEL) Greatest concentration or amount of a substance, found by experiment or observation, that causes no alterations of morphology, functional capacity, growth, development, or life span of target organisms distinguishable from those observed in normal (control) organisms of the same species and strain under the same defined conditions of exposure. Source: PAC, 1993, 65, 2003 (Glossary for chemists of terms used in toxicology (IUPAC Recommendations 1993)) on page 2076 nodal plane A plane of a system in which the value of the orbital wave-function equals zero. This plane defines a region of zero electron density for the orbital. Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1955 noise The random fluctuations occurring in a signal that are inherent in the combination of instrument and method. Source: PAC, 1989, 61, 1657 (Nomenclature for automated and mechanised analysis (Recommendations 1989)) on page 1663 See also: PAC, 1995, 67, 1745 (Nomenclature, symbols, units and their usage in spectrochemical analysis-XI. Detection of radiation (IUPAC Recommendations 1995)) on page 1750 IUPAC Compendium of Chemical Terminology 999 of 1622 nominal linear flow, in chromatography The volumetric flowrate of the mobile phase divided by the area of the cross section of the column ( ) i.e. the linear flowrate in a part of the column not containing packing. Source: Orange Book, p. 102 nominally labelled tracer A tracer in which the label is present mainly in a specified position. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2526 non-calorimetric thermophysical measurements These include ( , , ) measurements, saturated density measurements, and any other measurements which give information on the ( , , ) surface of a fluid. Source: PAC, 1981, 53, 1805 (Assignment and presentation of uncertainties of the numerical results of thermodynamic measurements (Provisional)) on page 1818 non-crossing rule Electronic states of the same symmetry cannot cross along a reaction coordinate. Note: Only applies to diatomic molecules and not to polyatomic molecules. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 374 non-crystalline electrodes Electrodes in which a support matrix, containing an ion exchanger (either cationic or anionic), a plasticizer solvent, and possibly an uncharged, selectivity-enhancing species, forms the ion-selective membrane which is usually interposed between two aqueous solutions. The support used can be either macroporous [e.g. poly(propylene carbonate) filter, glass frit, etc.] or microporous (e.g. 'thirsty' glass or inert polymeric material such as PVC) yielding with the ion-exchanger and the solvent a 'solidified' homogeneous mixture. These electrodes exhibit a response due to the presence of the ion-exchange material in the membrane. The solvent-polymeric-membrane is an example. IUPAC Compendium of Chemical Terminology 1000 of 1622 Source: PAC, 1994, 66, 2527 (Recommendations for nomenclature of ionselective electrodes (IUPAC Recommendations 1994)) on page 2534 non-destructive activation analysis An activation analysis procedure in which, after the irradiation, no chemical and physical operations are applied which cause a change of any properties of the sample. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1535 non-dissociative chemisorption See: chemisorption Source: PAC, 1976, 46, 71 (Manual of Symbols and Terminology for Physicochemical Quantities and Units - Appendix II. Definitions, Terminology and Symbols in Colloid and Surface Chemistry. Part II: Heterogeneous Catalysis) on page 76 non-draining An adjective describing a chain macromolecule that behaves in a hydrodynamic field as though the solvent within the domain of the macromolecule were virtually immobilized with respect to the macromolecule. Source: Purple Book, p. 61 non-equilibrium reaction If the reactants in a chemical reaction are not initially present in a Boltzmann distribution the reaction is referred to as a non-equilibrium reaction. See: equilibrium reaction Source: PAC, 1996, 68, 149 (A glossary of terms used in chemical kinetics, including reaction dynamics (IUPAC Recommendations 1996)) on page 175 non-graphitic carbon All varieties of solids consisting mainly of the element carbon with two-dimensional long-range order of the carbon atoms in planar hexagonal networks, but without any measurable crystallographic order in the third direction (c-direction) apart from more or less parallel stacking. Note: IUPAC Compendium of Chemical Terminology 1001 of 1622 Some varieties of non-graphitic carbon convert on heat treatment to graphitic carbon (graphitizable carbon) but some others do not (non-graphitizable carbon). See also: amorphous carbon Source: PAC, 1995, 67, 473 (Recommended terminology for the description of carbon as a solid (IUPAC Recommendations 1995)) on page 498 non-graphitizable carbon A non-graphitic carbon which cannot be transformed into graphitic carbon solely by high-temperature treatment up to under atmospheric pressure or lower pressure. Note: The term non-graphitizable is limited to the result of heat treatment without additional influence of foreign matter or neutron radiation. Non-graphitizable carbon can be transformed into graphitic carbon by a high-temperature process via intermediate dissolution in foreign matter and precipitation under high pressure or by radiation damage. Source: PAC, 1995, 67, 473 (Recommended terminology for the description of carbon as a solid (IUPAC Recommendations 1995)) on page 498 non-ideal, linear chromatography Chromatographic process, where the retention is governed by a linear distribution isotherm and normal peak-broadening takes place. Note: This case is commonly assumed in analytical chromatography. Source: PAC, 1996, 68, 1591 (Nomenclature for non-linear chromatography (IUPAC Recommendations 1996)) on page 1594 non-ideal, non-linear chromatography Chromatographic process, where both isotherm curvature and other peak-broadening processes (such as diffusion) contribute to the peak shape. Note: This case comprises most peaks in common practice that are characterised as 'tailing' or 'fronting'. Source: PAC, 1996, 68, 1591 (Nomenclature for non-linear chromatography (IUPAC Recommendations 1996)) on page 1594 non-ideal chromatography Chromatographic process with normal peak-broadening effects. IUPAC Compendium of Chemical Terminology 1002 of 1622 Source: PAC, 1996, 68, 1591 (Nomenclature for non-linear chromatography (IUPAC Recommendations 1996)) on page 1593 non-isotopic labelling Labelling in which the resulting product has a different chemical composition from the initial one. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2521 non-Kekulé molecules Molecules that are fully conjugated, but each of whose Kekulé structures contains at least two atoms that are not π-bonded. Examples of non-Kekule hydrocarbons are trimethylenemethane (2methylidenepropane-1,3-diyl) 1, m-benzodiquinomethane (benzene-1,2-diyldimethyl) 2, and 1,2,4,5 - tetramethylenebenzene (benzene-1,2,4,5-tetrayltetramethyl) 3. Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1955 non-linear chromatography Chromatographic process, where the retention is governed by a non-linear distribution isotherm. Source: PAC, 1996, 68, 1591 (Nomenclature for non-linear chromatography (IUPAC Recommendations 1996)) on page 1593 non-linear distribution isotherm in chromatography A distribution isotherm which is not linear. Note: A non-linear isotherm can have several shapes, as classified by Brunauer et al. In chromatography convex or concave shapes are common, as well as combinations. IUPAC Compendium of Chemical Terminology 1003 of 1622 Source: PAC, 1996, 68, 1591 (Nomenclature for non-linear chromatography (IUPAC Recommendations 1996)) on page 1593 non-linear optical effect An effect brought about by electromagnetic radiation the magnitude of which is not proportional to the irradiance. Non-linear optical effects of importance to photochemistry are harmonic frequency generation, lasers, Raman shifting, upconversion, and others. Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2256 non-linear optical polymer Polymer that exhibits an optical effect brought about by electromagnetic radiation such that the magnitude of the effect is not proportional to the irradiance. Notes: 1. See also: non-linear optical effect. 2. An example of non-linear optical effects is the generation of higher harmonics of the incident light wave. 3. A polymer that exhibits a non-linear optical effect due to anisotropic electric susceptibilities when subjected to electric field together with light irradiation is called an electro-optical polymer. A polymer that exhibits electro-optical behavior combined with photoconductivity is called a photo-refractive polymer. Source: PAC, 2004, 76, 889 (Definitions of terms relating to reactions of polymers and to functional polymeric materials (IUPAC Recommendations 2003)) on page 900 non-linear optical techniques Collective name applied to techniques related to non-linear optical effects. Notes: 1. Some of these spectroscopies are four-wave mixing, hole burning, photon echoes, timeresolved coherent anti-Stokes Raman (CARS), transient grating and stimulated pumping. These techniques differ in the nature of the pulse sequence, the geometry and the choice of a spatial direction (phase matching), as well as the mode of detection. 2. Low order non-linear techniques, such as three-wave mixing, are related to the second order polarization. For a random isotropic medium with inversion symmetry, the secondorder polarization vanishes and then the lowest order optical non-linear techniques, as well as the higher order non-linear techniques are related to the third-order polarization and the corresponding 'hyper-susceptibility'. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 374 IUPAC Compendium of Chemical Terminology 1004 of 1622 non-linearity error in spectrochemical analysis An error caused by any deviation from linearity of the response of the detector to the measured radiant power. Source: PAC, 1988, 60, 1449 (Nomenclature, symbols, units and their usage in spectrochemical analysis VII. Molecular absorption spectroscopy, ultraviolet and visible (UV/VIS) (Recommendations 1988)) on page 1456 non-polarized interphases Interphases for which the exchange of common charged components between the phases proceeds unhindered. Source: PAC, 1983, 55, 1251 (Interphases in systems of conducting phases (Provisional)) on page 1253 non-radiative decay The disappearance of an excited species due to a radiationless transition. Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2256 non-specific adsorption Ions approach an interface differently depending on the forces in play. Ions are non-specifically adsorbed (positively or negatively) when they are subjected in the interphase only to long-range coulombic interactions (attraction or repulsion). They are believed to retain their solvation shell, and in the position of closest approach to the interface they are separated from it by one or more molecular layers. See also: specific adsorption Source: PAC, 1986, 58, 437 (Interphases in systems of conducting phases (Recommendations 1985)) on page 447 non-uniform corrosion Corrosion is non-uniform if the time average of the corrosion current through a unit area depends on its position on the surface. Non-uniform corrosion can be due to inhomogeneities of structure or of composition of the corroding material, or to inhomogeneities of the environment. Special cases of non-uniform corrosion such as pitting or intergranular corrision are sometimes called localized corrosion. IUPAC Compendium of Chemical Terminology 1005 of 1622 Source: PAC, 1989, 61, 19 (Electrochemical corrosion nomenclature (Recommendations 1988)) on page 21 non-uniform polymer A polymer comprising molecules non-uniform with respect to relative molecular mass or constitution or both. See: uniform polymer (3) Source: PAC, 1996, 68, 2287 (Glossary of basic terms in polymer science (IUPAC Recommendations 1996)) on page 2302 non-vertical energy transfer Energy transfer process with a low Franck–Condon factor because the nuclear geometries of the minima on the ground- and excited-state potential energy surfaces of the donor or of the acceptor are strongly displaced. Note: A Franck–Condon transition is always vertical. Thus, it is not strictly correct to speak about non-vertical transition. However, the term implies that the projection of all the vibrational wave functions of the donor state onto all the vibrational functions of the acceptor will result in low Franck– Condon factors. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 374 nonadiabatic coupling This is momentum coupling between two adiabatic potential-energy surfaces. See also: adiabatic Source: PAC, 1996, 68, 149 (A glossary of terms used in chemical kinetics, including reaction dynamics (IUPAC Recommendations 1996)) on page 175 nonadiabatic electron transfer See: diabatic electron transfer Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2255 nonadiabatic photoreaction [obsolete] Synonymous with diabatic photoreaction. Use of the double negative is discouraged. IUPAC Compendium of Chemical Terminology 1006 of 1622 Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2256 nonbonded interactions Intramolecular attractions or repulsions between atoms that are not directly linked to each other, affecting the thermodynamic stability of the chemical species concerned. See also: eclipsing strain, transannular strain Source: PAC, 1996, 68, 2193 (Basic terminology of stereochemistry (IUPAC Recommendations 1996)) on page 2212 nonbonding molecular orbital A molecular orbital whose occupation by electrons does not contribute (or contributes insignificantly) to the binding energy of the molecule. Generally, a nonbonding MO represents the highest occupied molecular orbital of a molecule. Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1955 nonclassical carbocation A carbocation the ground state of which has delocalized (bridged) bonding π- or σ-electrons. (N.B. Allylic and benzylic carbocations are not considered nonclassical.) Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1145 nonclassical structure The structure of molecules or molecular ions that escapes description in terms of conventional rules of valency and stereochemistry. Nonclassical structures are characteristic of carbonium ions with hypercoordinated (see hypercoordination) carbon atoms, e.g., methanium ion 1, pyramidal dication C6H62+ 2 (isomeric to benzene dication), and the molecular species whose structure cannot be adequately represented by the equilibrium (2-norbornyl cation, 3) or resonance of two or more classical structures. From the stereochemical point of view, those structures are assigned to the nonclassical type for which all tetracoordinate carbon bonds extend into a single hemisphere, i.e., the valence angle of a carbon atom is greater than 180°. A hypothetical example is pyramidane, 4, the structure of which corresponds to a local minimum on the C5H4 potential energy surface. IUPAC Compendium of Chemical Terminology 1007 of 1622 Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1955 nonselectively labelled An isotopically labelled compound is designated as nonselectively labelled when the position(s) and the number of the labelling nuclide(s) are both undefined. Source: Blue Book, p. 517 PAC, 1981, 53, 1887 (Nomenclature of inorganic chemistry: II.1 - Isotopically modified compounds) on page 1896 norAffix used to denote the elimination of one methylene group from a side chain of a parent structure (including a methyl group). Source: Blue Book, p. 497 normal The term 'normal' in e.g. 'normal boiling temperature' means the value at a pressure of. Source: PAC, 1994, 66, 533 (Standard quantities in chemical thermodynamics. Fugacities, activities and equilibrium constants for pure and mixed phases (IUPAC Recommendations 1994)) on page 537 normal distribution See: Gaussian band shape Source: PAC, 1990, 62, 2167 (Glossary of atmospheric chemistry terms (Recommendations 1990)) on page 2203 IUPAC Compendium of Chemical Terminology 1008 of 1622 normal kinetic isotope effect See: isotope effect Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1145 normal region (for electron transfer) Region for which the rate constants increase with increasing exergonicity of the reaction in plots relating rate constants for electron transfer, or quantities related to it, with the standard Gibbs energy for the reaction ( ). This region is predicted by the Marcus theory for outer-sphere electron transfer for the case of in the Marcus equation. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 375 normal stress, Force acting normally to a surface divided by the area of the surface. Source: Green Book, 2nd ed., p. 12 normal X-ray level Synonymous with diagram level. Source: PAC, 1991, 63, 735 (Nomenclature, symbols, units and their usage in spectrochemical analysis - VIII. Nomenclature system for X-ray spectroscopy (Recommendations 1991)) on page 738 normal-phase chromatography An elution procedure in which the stationary phase is more polar then the mobile phase. This term is used in liquid chromatography to emphasize the contrast to reversed-phase chromatography. Source: PAC, 1993, 65, 819 (Nomenclature for chromatography (IUPAC Recommendations 1993)) on page 826 IUPAC Compendium of Chemical Terminology 1009 of 1622 normalization A mathematical procedure which ensures that the integral of the square of modulus of a wave-function over all space equals 1. The constant required to ensure that a wave-function is normalized is termed the normalization constant. Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1956 Norrish Type I photoreaction α-Cleavage of an excited carbonyl compound leading to an acyl-alkyl radical pair (from an acyclic carbonyl compound) or an acyl-alkyl diradical (from a cyclic carbonyl compound) as a primary photoproduct. Notes: 1. An example is: 2. Decarbonylation of the primary diradical formed from cyclic carbonyl compounds and subsequent recombination of the biradical afford very interesting cyclic compounds. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 375 Norrish Type II photoreaction Intramolecular abstraction of a γ-hydrogen by an excited carbonyl compound to produce a 1,4diradical as a primary photoproduct. Note: An example is: Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 375 IUPAC Compendium of Chemical Terminology 1010 of 1622 Norrish–Yang reaction Photochemical process leading to the formation of cyclobutanols from excited carbonyl compounds by intramolecular γ-hydrogen abstraction (Norrish Type II photoreaction) and subsequent 1,4diradical combination. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 375 nth order phase transition A transition in which the molar Gibbs energy (or chemical potential) and its th-order derivatives are continuous, whereas the th-order derivatives with respect to temperature and pressure are discontinuous at the transition point. See: first-order transition, second-order transition Source: PAC, 1994, 66, 577 (Definitions of terms relating to phase transitions of the solid state (IUPAC Recommendations 1994)) on page 587 nuclear atom See: coordination entity Source: Red Book, p. 145 Blue Book, p. 337 nuclear chemistry The part of chemistry which deals with the study of nuclei and nuclear reactions using chemical methods. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2516 nuclear decay A spontaneous nuclear transformation. IUPAC Compendium of Chemical Terminology 1011 of 1622 Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2518 nuclear disintegration Nuclear decay involving a splitting into more nuclei or the emission of particles. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1540 nuclear fission The division of a nucleus into two or more parts with masses of equal order of magnitude, usually accompanied by the emission of neutrons, gamma radiation and, rarely, small charged nuclear fragments. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2519 nuclear fuel Material containing fissile nuclides, which when placed in a reactor, enables a chain reaction to be achieved. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1543 nuclear fusion See: nuclear fusion reaction Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 nuclear fusion reaction A reaction between two light nuclei resulting in the production of a nuclear species heavier than either initial nucleus. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1543 IUPAC Compendium of Chemical Terminology 1012 of 1622 nuclear graphite A polygranular graphite material for use in nuclear reactor cores consisting of graphitic carbon of very high chemical purity. High purity is needed to avoid absorption of low-energy neutrons and the production of undesirable radioactive species. Note: Apart from the absence of neutron-absorbing impurities, modern reactor graphites are also characterized by a high degree of graphitization and no preferred bulk orientation. Such properties increase the dimensional stability of the nuclear graphite at high temperatures and in a high flux of neutrons. The term nuclear graphite is often, but incorrectly, used for any graphite material in a nuclear reactor, even if it serves only for structural purposes. Source: PAC, 1995, 67, 473 (Recommended terminology for the description of carbon as a solid (IUPAC Recommendations 1995)) on page 498 nuclear isomers Nuclides having the same mass number and atomic number, occupying different nuclear energy states. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1545 nuclear level One of the energy values at which a nucleus can exist for an appreciable time ( ). Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 nuclear magneton Electromagnetic fundamental physical constant where is the electron rest mass, the proton rest mass and , the Bohr magneton. Source: CODATA Bull. 1986, 63, 1 nuclear particle A nucleus or any of its constituents in any of their energy states. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 IUPAC Compendium of Chemical Terminology 1013 of 1622 nuclear quadrupole moment (spectroscopic) A parameter which describes the effective shape of the equivalent ellipsoid of the nuclear charge distribution, for prolate (e.g. 57Fe, 197Au); for oblate (e.g. 119Sn, 129I) nuclei. Source: PAC, 1976, 45, 211 (Nomenclature and Conventions for Reporting Mossbauer Spectroscopic Data) on page 214 nuclear reactor A device in which a self-sustaining nuclear fission chain reaction can be maintained and controlled. The term is sometimes applied to a device in which a nuclear fusion reaction can be produced and controlled. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1550 nuclear transformation The change of one nuclide into another with a different proton number or nucleon number. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1553 nuclear transition For a nucleus a change from one quantized energy state into another or a nuclear transformation. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1553 nuclearity The number of central atoms joined in a single coordination entity by bridging ligands or metal-metal bonds is indicated by dinuclear, trinuclear, tetranuclear, polynuclear, etc. Source: PAC, 1997, 69, 1251 (Glossary of terms used in bioinorganic chemistry (IUPAC Recommendations 1997)) on page 1288 nucleating agent A material either added to or present in a system, which induces either homogeneous or heterogeneous nucleation. IUPAC Compendium of Chemical Terminology 1014 of 1622 Source: PAC, 1972, 31, 577 (Manual of Symbols and Terminology for Physicochemical Quantities and Units, Appendix II: Definitions, Terminology and Symbols in Colloid and Surface Chemistry) on page 608 nucleation and growth A process in a phase transition in which nuclei of a new phase are first formed, followed by the propagation of the new phase at a faster rate. See: continuous precipitation, discontinuous precipitation Source: PAC, 1994, 66, 577 (Definitions of terms relating to phase transitions of the solid state (IUPAC Recommendations 1994)) on page 587 nucleation Also contains definition of: simultaneous nucleation in colloid chemistry The process by which nuclei are formed in solution. The condensation of a single chemical compound is called homogeneous nucleation. The simultaneous condensation of more than one compound is called simultaneous nucleation. The condensation of a compound on a foreign substance is called heterogeneous nucleation. Source: Orange Book, p. 84 See also: PAC, 1972, 31, 577 (Manual of Symbols and Terminology for Physicochemical Quantities and Units, Appendix II: Definitions, Terminology and Symbols in Colloid and Surface Chemistry) on page 608 nucleation of phase separation in polymer chemistry Initiation of phase domain formation through the presence of heterogeneities. Note: In a metastable region of a phase diagram, phase separation is initiated only by nucleation. Source: PAC, 2004, 76, 1985 (Definition of terms related to polymer blends, composites, and multiphase polymeric materials (IUPAC Recommendations 2004)) on page 1997 nucleic acids Macromolecules, the major organic matter of the nuclei of biological cells, made up of nucleotide units, and hydrolysable into certain pyrimidine or purine bases (usually adenine, cytosine, guanine, thymine, uracil), D-ribose or 2-deoxy-D-ribose and phosphoric acid. IUPAC Compendium of Chemical Terminology 1015 of 1622 See: ribonucleic acids, , deoxyribonucleic acids Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 See also: White Book, p. 110 PAC, 1992, 64, 143 (Glossary for chemists of terms used in biotechnology (IUPAC Recommendations 1992)) on page 161 nucleofuge A leaving group that carries away the bonding electron pair. For example, in the hydrolysis of an alkyl chloride, Cl− is the nucleofuge. The tendency of atoms or groups to depart with the bonding electron pair is called nucleofugality. The adjective is nucleofugal. See also: electrofuge, nucleophile Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1145 nucleon Heavy nuclear particle: proton or neutron. Source: Physical Chemistry Division, unpublished nucleon number Synonymous with mass number. Source: Green Book, 2nd ed., p. 20 IUPAC Compendium of Chemical Terminology 1016 of 1622 nucleophile (nucleophilic) A nucleophile (or nucleophilic reagent) is a reagent that forms a bond to its reaction partner (the electrophile) by donating both bonding electrons. A 'nucleophilic substitution reaction' is a heterolytic reaction in which the reagent supplying the entering group acts as a nucleophile. For example: The term 'nucleophilic' is also used to designate the apparent polar character of certain radicals, as inferred from their higher relative reactivity with reaction sites of lower electron density. Nucleophilic reagents are Lewis bases. Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1146 nucleophilic aromatic photosubstitution Photosubstitution in aromatic compounds with the participation of nucleophiles. Note: While electrophilic substitution is the most common reaction on aromatic compounds in the ground state, nucleophilic substitution is typical for the excited states of these compounds. Some electrophilic aromatic photosubstitutions (upon excitation of the aromatic substrate) have been reported. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 376 nucleophilic catalysis Catalysis by a Lewis base, involving formation of a Lewis adduct as a reaction intermediate. For example, the hydrolysis of acetic anhydride in aqueous solution catalysed by pyridine: See also: electrophilic, nucleophilicity IUPAC Compendium of Chemical Terminology 1017 of 1622 Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1146 nucleophilicity 1. 2. The property of being nucleophilic. The relative reactivity of a nucleophilic reagent. (It is also sometimes referred to as 'nucleophilic power'.) Qualitatively, the concept is related to Lewis basicity. However, whereas Lewis basicity is measured by relative equilibrium constants: nucleophilicity of a Lewis base is measured by relative rate constants of different nucleophilic reagents towards a common substrate, most commonly involving formation of a bond to carbon: See also: electrophilicity, Ritchie equation, Swain–Scott equation Source: PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1146 nucleoproteins Proteins having nucleic acids as prosthetic groups, and thus yielding nucleic acids (or their cleavage products) as well as amino acids on hydrolytic cleavage. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 nucleosides Ribosyl or deoxyribosyl derivatives (rarely, other glycosyl derivatives) of certain pyrimidine or purine bases. They are thus glycosylamines or N-glycosides related to nucleotides by the lack of phosphorylation. It has also become customary to include among nucleosides analogous substances in which the glycosyl group is attached to carbon rather than nitrogen ('C-nucleosides'). See also: nucleic acids Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 See also: White Book, p. 110 IUPAC Compendium of Chemical Terminology 1018 of 1622 PAC, 1992, 64, 143 (Glossary for chemists of terms used in biotechnology (IUPAC Recommendations 1992)) on page 161 nucleotide bases The heterocyclic pyrimidine and purine compounds which are constituents of all nucleic acids. Adenine (A), guanine (G) and cytosine (C) are found in both DNA and RNA, thymine (T) is found (primarily) in DNA and uracil (U) only in RNA. Source: PAC, 1992, 64, 143 (Glossary for chemists of terms used in biotechnology (IUPAC Recommendations 1992)) on page 147 nucleotides Compounds formally obtained by esterification of the 3 or 5 hydroxy group of nucleosides with phosphoric acid. They are the monomers of nucleic acids and are formed from them by hydrolytic cleavage. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1352 See also: White Book, p. 109 White Book, p. 111 PAC, 1992, 64, 143 (Glossary for chemists of terms used in biotechnology (IUPAC Recommendations 1992)) on page 162 nucleus The positively charged central portion of an atom, excluding the orbital electrons. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 IUPAC Compendium of Chemical Terminology 1019 of 1622 nuclide A species of atom, characterized by its mass number, atomic number and nuclear energy state, provided that the mean life in that state is long enough to be observable. Source: PAC, 1994, 66, 2513 (Nomenclature for radioanalytical chemistry (IUPAC Recommendations 1994)) on page 2522 Red Book, p. 35 Orange Book, p. 233 PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 nuclidic mass The rest mass of a nuclide expressed in atomic mass units. Source: PAC, 1982, 54, 1533 (Glossary of terms used in nuclear analytical chemistry (Provisional)) on page 1547 nuisance threshold in atmospheric chemistry That concentration of an air pollutant that is considered objectionable. In the case of a substance with an objectionable odour, it is the smallest concentration of the substance which can be detected by a human being (nose). Source: PAC, 1990, 62, 2167 (Glossary of atmospheric chemistry terms (Recommendations 1990)) on page 2203 number concentration, , Number of entities of a constituent in a mixture divided by the volume of the mixture. See: concentration Source: Green Book, 2nd ed., p. 39 PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 984 number content, Number of defined particles, or elementary entities, of a component in a system divided by the mass of that system. IUPAC Compendium of Chemical Terminology 1020 of 1622 Source: PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 984 number density, Number of particles divided by the volume they occupy. Source: Green Book, 2nd ed., p. 39 PAC, 1990, 62, 2167 (Glossary of atmospheric chemistry terms (Recommendations 1990)) on page 2183 number flow rate, Number of defined particles, or elementary entities of a defined component, crossing a cross-section divided by the time. See: mass flow rate, volume flow rate Source: PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 984 number fraction See: amount fraction Source: Green Book, 2nd ed., p. 41 PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 984 number of entities, Integer number obtained by counting of entities, which are usually molecules atoms or ions. Source: Green Book, 2nd ed., p. 39 PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 985 number-distribution function A distribution function in which the relative amount of a portion of a substance with a specific value, or a range of values, of the random variable(s) is expressed in terms of mole fraction. Source: Purple Book, p. 56 IUPAC Compendium of Chemical Terminology 1021 of 1622 numerical value of a quantity Quotient of the value of a quantity and the unit used in its expression. See: value of a quantity Source: PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 985 observation height, in flame emission and absorption spectrometry The difference in height between the axis of the observed space (optical axis) and the burner tip (in ). The optical axis of the instrument should be the same as the optical axis of the observation space. Other definitions which express the observation height as a fraction of the whole flame height are not recommended because the height to the tip of the flame is not well defined. The observation height should be stated in the method. Source: PAC, 1986, 58, 1737 (Quantities and units in clinical chemistry: Nebulizer and flame properties in flame emission and absorption spectrometry (Recommendations 1986)) on page 1741 observation pathlength, in flame emission and absorption spectrometry The intersection of the optical axis and the observation space (in ). Source: PAC, 1986, 58, 1737 (Quantities and units in clinical chemistry: Nebulizer and flame properties in flame emission and absorption spectrometry (Recommendations 1986)) on page 1741 observation space in flame emission and absorption spectrometry The intersection of the optical beam and that part of the flame where the net signal is at least half of the maximum net signal. The characteristics of the observation space depend on the temperature of the flame, the stoichiometry of the gases and the properties of the processed fluid (presence of proteins for instance). The observation space in atomic absorption is analogous to the observation space of a cuvette in molecular absorption spectrometry. See also: observation volume Source: PAC, 1986, 58, 1737 (Quantities and units in clinical chemistry: Nebulizer and flame properties in flame emission and absorption spectrometry (Recommendations 1986)) on page 1741 IUPAC Compendium of Chemical Terminology 1022 of 1622 observation volume, in flame emission and absorption spectrometry The volume of that part of the flame that is observed through the optical device (in See also: observation space ). Source: PAC, 1986, 58, 1737 (Quantities and units in clinical chemistry: Nebulizer and flame properties in flame emission and absorption spectrometry (Recommendations 1986)) on page 1741 occlusion (molecular) The process of incorporation of foreign substances as molecular species within precipitates as they are formed. Source: Orange Book, p. 85 octahedro- in inorganic nomenclature An affix used in names to denote six atoms bound into an octahedron. Source: Red Book, p. 245 Blue Book, p. 465 odd-electron ion Synonymous with radical ion. Source: PAC, 1991, 63, 1541 (Recommendations for nomenclature and symbolism for mass spectroscopy (including an appendix of terms used in vacuum technology). (Recommendations 1991)) on page 1549 ODMR (Optically Detected Magnetic Resonance) Also contains definitions of: ADMR, DEDMR, DFDMR, PDMR, RYDMR A double resonance technique in which transitions between spin sublevels are detected by optical means. Usually these are sublevels of a triplet and the transitions are induced by microwaves. For different types of optical detection the following expressions are used: ADMR (absorption), DEDMR (delayed emission, non-specified), DFDMR (delayed fluorescence), FDMR (fluorescence), PDMR (phosphorescence). If a reaction yield is followed the expression RYDMR (reaction yield detected magnetic resonance) is used. Source: PAC, 1996, 68, 2223 (Glossary of terms used in photochemistry (IUPAC Recommendations 1996)) on page 2257 IUPAC Compendium of Chemical Terminology 1023 of 1622 odour threshold in atmospheric chemistry The concentration of a compound which produces an odour which is detectable by a human being (nose). For certain compounds this threshold is very low (e.g. 1 part in for certain sulfides). Source: PAC, 1990, 62, 2167 (Glossary of atmospheric chemistry terms (Recommendations 1990)) on page 2204 ohm SI derived unit of electrical resistance,. Source: Green Book, 2nd ed., p. 72 PAC, 1996, 68, 957 (Glossary of terms in quantities and units in Clinical Chemistry (IUPAC-IFCC Recommendations 1996)) on page 985 OLED Acronym for organic light-emitting device. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 377 olefins Acyclic and cyclic hydrocarbons having one or more carbon–carbon double bonds, apart from the formal ones in aromatic compounds. The class olefins subsumes alkenes and cycloalkenes and the corresponding polyenes. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 oligo A prefix meaning 'a few', and used for compounds with a number of repeating units intermediate between those in monomers and those in high polymers. The limits are not precisely defined, and in practice vary with the type of structure being considered, but are generally from 3 to 10, e.g. oligopeptides, oligosaccharides. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 IUPAC Compendium of Chemical Terminology 1024 of 1622 oligomer Also contains definition of: telomer A substance composed of oligomer molecules. An oligomer obtained by telomerization is often termed a telomer. Source: PAC, 1996, 68, 2287 (Glossary of basic terms in polymer science (IUPAC Recommendations 1996)) on page 2289 oligomer molecule Also contains definition of: oligomeric A molecule of intermediate relative molecular mass, the structure of which essentially comprises a small plurality of units derived, actually or conceptually, from molecules of lower relative molecular mass. Notes: 1. A molecule is regarded as having an intermediate relative molecular mass if it has properties which do vary significantly with the removal of one or a few of the units. 2. If a part or the whole of the molecule has an intermediate relative molecular mass and essentially comprises a small plurality of units derived, actually or conceptually, from molecules of lower relative molecular mass, it may be described as oligomeric, or by oligomer used adjectivally. Source: PAC, 1996, 68, 2287 (Glossary of basic terms in polymer science (IUPAC Recommendations 1996)) on page 2289 oligomerization The process of converting a monomer or a mixture of monomers into an oligomer. An oligomerization by chain reaction carried out in the presence of a large amount of chain-transfer agent, so that the endgroups are essentially fragments of the chain transfer agent, is termed telomerization. Source: PAC, 1996, 68, 2287 (Glossary of basic terms in polymer science (IUPAC Recommendations 1996)) on page 2306 oligonucleotides See: oligo, nucleotides Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 See also: White Book, p. 111 IUPAC Compendium of Chemical Terminology 1025 of 1622 oligopeptides See: oligo, peptides Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 See also: White Book, p. 48 oligosaccharides See: oligo, saccharides Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 one-bond-flip Acronym: OBF Photoisomerization of one double bond in a conjugated double bond system. It is a regular E/Z photoisomerization. Compare with bicycle-pedal mechanism, hula-twist mechanism. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 377 one-photon photochromism Photochromic process involving a one-photon mechanism. In this case, the photoproduct B is formed from the singlet or from the triplet state of the thermodynamically-stable molecular entity A. Source: PAC, 2007, 79, 293 (Glossary of terms used in photochemistry, 3rd edition (IUPAC Recommendations 2006)) on page 377 IUPAC Compendium of Chemical Terminology 1026 of 1622 onion morphology Multiphase morphology of roughly spherical shape that comprises alternating layers of different polymers arranged concentrically, all layers being of similar thickness. Source: PAC, 2004, 76, 1985 (Definition of terms related to polymer blends, composites, and multiphase polymeric materials (IUPAC Recommendations 2004)) on page 2004 onium compounds 1. Cations (with their counter-ions) derived by addition of a hydron to a mononuclear parent hydride of the nitrogen, chalcogen and halogen families. (H4N+) ammonium (H3O+) oxonium (H2F+) fluoronium (H4P+) phosphonium (H3S+) sulfonium (H2Cl+) chloronium (H4As+) arsonium (H3Se+) selenonium (H2Br+) bromonium (H4Sb+) stibonium (H3Te+) telluronium (H2I+) iodonium 2. (H4Bi+) bismuthonium Derivatives formed by substitution of the above parent ions by univalent groups. The number of substituted hydrogen atoms is, especially in the case of hydrocarbyl substituents, indicated by the adjectives primary, secondary, tertiary or quaternary. E.g. (Cl2F+) dichlorofluoronium, (CH3)2S+H dimethylsulfonium (a secondary sulfonium ion), Cl(CH3)3P+ 3. chlorotrimethylphosphonium, (CH3CH2)4N+ tetraethylammonium (a quaternary ammonium ion). See also: arsonium compounds, halonium ions, oxonium ions, phosphonium compounds, quaternary ammonium compounds, stibonium compounds, sulfonium compounds Derivatives formed by substitution of the above parent ions by groups having two or three free valencies on the same atom. Such derivatives are, where possible, designated by a specific class name. E.g. RC≡O+ hydrocarbylidyne oxonium ions, R2C=N+H2X− iminium compounds, RC≡NH+ nitrilium ions. Source: PAC, 1995, 67, 1307 (Glossary of class names of organic compounds and reactivity intermediates based on structure (IUPAC Recommendations 1995)) on page 1353 IUPAC Compendium of Chemical Terminology 1027 of 1622 PAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1146 open film Also contains definitions of: closed film, partly open film A film in which mass transfer can occur between the film and the coexisting bulk phases, for all the components. The term partly open film applies to a film in which mass transfer can occur only for certain components. The term closed film applies to those films with fixed mass. Source: PAC, 1994, 66, 1667 (Thin films including layers: terminology in relation to their preparation and characterization (IUPAC Recommendations 1994)) on page 1671 open hearth furnace in atmospheric chemistry Reverberatory furnace, containing a basin-shaped hearth, for melting and refining suitable types of pig iron, iron ore, and scrap for steel production. A large amount of dust from ore and other materials and splashings from slag are carried away by the waste gases; a supplementary chamber is commonly used for collecting slag and dust. Source: PAC, 1990, 62, 2167 (Glossary of atmospheric chemistry terms (Recommendations 1990)) on page 2204 open-shell systems Atomic or molecular systems in which the electrons are not completely assigned to orbitals in pairs. Source: PAC, 1999, 71, 1919 (Glossary of terms used in theoretical organic chemistry) on page 1956 open-tubular column in chromatography A column, usually having a small diameter, in which either the inner tube wall, or a liquid or active solid held stationary on the tube wall acts as the stationary phase and there is an open, unrestricted path for the mobile phase. Source: PAC, 1993, 65, 819 (Nomenclature for chromatography (IUPAC Recommendations 1993)) on page 831 Orange Book, p. 97 IUPAC Compendium of Chemical Terminology 1028 of 1622 operational pH cell An electrochemical cell which is the basis of practical pH measurement consisting of a hydrogen ionresponsive electrode (hydrogen gas, or glass) and a reference electrode immersed in the test solution.	1,008
https://openalex.org/W2782035106	OpenAlex	Open Science	CC-By	2,018	Air-breathing behavior, oxygen concentrations, and ROS defense in the swimbladders of two erythrinid fish, the facultative air-breathing jeju, and the non-air-breathing traira during normoxia, hypoxia and hyperoxia	Bernd Pelster	English	Spoken	11,274	20,367	Journal of Comparative Physiology B (2018) 188:437–449 https://doi.org/10.1007/s00360-017-1142-1 Journal of Comparative Physiology B (2018) 188:437–449 https://doi.org/10.1007/s00360-017-1142-1 ORIGINAL PAPER ORIGINAL PAPER Abstract The jeju Hoplerythrinus unitaeniatus and the traira Hoplias malabaricus are two neighboring genera from the family of erythrinid fish, both possessing a two-chambered physostomous swimbladder. In the jeju the anterior section of the posterior bladder is highly vascularized, and the swimbladder is used for aerial respiration; the traira, in turn, is a water-breather that uses the swimbladder as a buoyancy organ and not for aerial oxygen uptake. Measurement of swimbladder oxygen partial pressure (PO2) of fish kept at 26 °C in normoxic, hyperoxic (28–32 mg O2 L− 1) or hypoxic (1–1.5 mg O2 L− 1) water revealed constant values in traira swimbladder. Under normoxic conditions in the jeju swimbladder PO2 was higher than in traira, and the PO2 significantly increased under hyperoxic conditions, even in the absence of air breathing. In jeju, air-breathing activity increased significantly under hypoxic conditions. Hypoxic air-breathing activity was negatively correlated to swimbladder PO2, indicating that the swimbladder was intensely used for gas exchange under these conditions. In traira, the capacity of the ROS defense system, as assessed by measurement of activities of enzymes involved in ROS degradation and total glu- tathione (GSH + GSSG) concentration, was elevated after 4 h of hyperoxic and/or hypoxic exposure, although swimbladder PO2 was not affected. In jeju, experiencing a higher variability in swimbladder PO2 due to the air-breathing activity, only a reduced responsiveness of the ROS defense system to changing environmental PO2 was detected. Keywords Air-breathing fish · ROS defense · Swimbladder · Hypoxia · Hyperoxia * Bernd Pelster bernd.pelster@uibk.ac.at Air-breathing behavior, oxygen concentrations, and ROS defense in the swimbladders of two erythrinid fish, the facultative air- breathing jeju, and the non-air-breathing traira during normoxia, hypoxia and hyperoxia Bernd Pelster1,2 · Chris M. Wood3 · Ellen Jung3 · Adalberto L. Val4 Received: 5 October 2017 / Revised: 11 December 2017 / Accepted: 19 December 2017 / Published online: 3 January 2018 © The Author(s) 2018. This article is an open access publication Communicated by G. Heldmaier. 4 Instituto Nacional de Pesquisas da Amazônia, Manaus, Brazil Introduction Indeed, in a recent study on erythrinid fishes, we demonstrated that there was a higher ROS defense capacity not only in the swimbladder tissue but also in the muscle tissue, of the facultative air-breathing jeju (Hoplerythrinus unitaeniatus) relative to its non-air- breathing close relative, the traira (Hoplias malabaricus) (Pelster et al. 2016). The jeju uses its swimbladder as an air-breathing organ, whereas the traira uses the swimblad- der only as a buoyancy organ. In non-ventilated gas cavities such as the traira swimbladder, inert gases such as nitrogen accumulate (Piiper 1965), so that PO2 in traira swimbladder is expected to be much lower than in the jeju swimbladder. GSH in its reduced form, and glutathione reductase (GR) subsequently reduces GSSG back to GSH at the expense of NADPH2 (Hermes-Lima 2004). Oxidized NADP+ may then be reduced again by the activity of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, enzymes of the pentose phosphate shunt.i By exposing fish to variable oxygen conditions (hypoxia as well as hyperoxia), it has been shown that the ROS defense system is quite flexible. Hyperoxic exposure not only induces an increased activity of ROS-degrading enzymes (Lushchak et al. 2005a; Lushchak and Bagnyu- kova 2006; Lushchak 2011) but also hypoxia often results in an increase in the activity of these enzymes (Lushchak et al. 2001, 2005b; Wilhelm Filho et al. 2005; Lushchak and Bagnyukova 2007; Tripathi et al. 2013). The latter may be the response to an increase in ROS under hypoxic conditions, but it may also be a preparation of the organ- ism for oxidative stress encountered during recovery from hypoxia, and the concept of a preparation for oxidative stress is widely accepted (Welker et al. 2013). In the present study, we, therefore, hypothesized that the ROS defense system of the two erythrinids, jeju (Hoplerythrinus unitaeniatus) and traira (Hoplias malabaricus), would also show this flexibil- ity to changing oxygen availability. We also hypothesized that the swimbladder of the air-breathing jeju, depending on the air-breathing activity, is frequently facing chang- ing oxygen partial pressures, while the swimbladder of the non-air-breathing traira was expected to face more or less constant low-oxygen partial pressures. Based on the results obtained, we intended to assess the possible contribution of the swimbladder to gas exchange. While the connection between hyperoxia and the gen- eration of ROS has been unequivocal, the relation between hypoxic conditions and the formation of ROS has been debated intensively. Introduction It was originally expected that the reduction in oxygen availability would result in a concomi- tant decrease in ROS production because oxygen is required for the generation of ROS (Welker et al. 2013). Meanwhile, several studies have shown, however, that reduced oxygen availability (hypoxia) may also cause oxidative stress (Wel- ker et al. 2013). In fish, hypoxia-induced oxidative stress has been detected in several species, including goldfish Caras- sius auratus (Lushchak et al. 2001), common carp Cypri- nus carpio (Lushchak et al. 2005b), rotan Perccottus glenii (Lushchak and Bagnyukova 2007), medaka (Oryzias latipes) (Lushchak and Bagnyukova 2007; Oehlers et al. 2007), pia- para (Leporinus elongatus) (Wilhelm Filho et al. 2005) and the Indian catfish Clarias batrachus (Tripathi et al. 2013). Materials and methods Experiments were performed at the Instituto Nacional de Pesquisas da Amazônia (INPA). All procedures were in compliance with Brazilian national and INPA animal care regulations. Erythrinid fish used for this study, the jeju and the traira, were caught by INPA fishermen, brought to the INPA and kept in outdoor fish tanks, supplied with running INPA freshwater. The water was continuously aerated. In the tanks, the fish had free access to air and could, therefore, breathe air voluntarily. Fish were fed daily until the day before the experiment. Water pH was 6.0 ± 0.05, the temperature was 26 ± 1 °C. i To prevent tissue damage in situations of variable oxygen availability when there is the danger of inordinate accumula- tion of ROS, animals have developed a defense system for rapidly breaking down ROS (Storey 1996; Wilhelm Filho 1996). This system consists of low molecular weight anti- oxidants such as glutathione (GSH + GSSG), thioredoxin (Trx-(SH2)/Trx-SS-Trx), ascorbic acid (vitamin C), retinol (vitamin A) or α-tocopherol (vitamin E). In addition, sev- eral enzymes can remove ROS, such as catalase and super- oxide dismutase (SOD). SOD catalyzes the dismutation of O2 − into O2 and H2O2, and catalase decomposes H2O2. Glu- tathione peroxidase (GPx) also decomposes H2O2 using the Introduction demand during periods of aquatic hypoxia, many fish of the Amazon Basin rely on aquatic surface respiration or even use specific tissues or organs for aerial gas exchange (Val and Almeida-Val 1995). In water-breathing fish arterial PO2 typically is well below aerial PO2 (Gilmour and Perry 1994; Kristensen et al. 2010). Epithelia exposed to air in these air breathing organs therefore face much higher PO2 values than tissues of water-breathing fish. Depending on the air-breath- ing activity and the removal of oxygen from the engulfed air, also quite variable PO2 values may be encountered in air-breathing organs. The oxygen concentration in the Amazon Basin is known to vary greatly, depending on season and the daily cycle (Val and Almeida-Val 1995; Muusze et al. 1998; Diaz and Breitburg 2009; Welker et al. 2013). To supplement oxygen Communicated by G. Heldmaier. Communicated by G. Heldmaier. High oxygen concentrations and especially hyperoxic conditions result in the generation of reactive oxygen spe- cies (ROS), not only in mammals but also in fish. Exposure to hyperoxia caused oxidative stress in different tissues of goldfish (brain, liver, kidney, muscle), as evidenced by an increase in protein carbonylation, TBARS (a byproduct of lipid peroxidation) and/or lipid peroxides themselves 1 Institut für Zoologie, Leopold-Franzens-Universität Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria 2 Center for Molecular Biosciences, University Innsbruck, Innsbruck, Austria 3 Department of Zoology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada 4 Instituto Nacional de Pesquisas da Amazônia, Manaus, Brazil (0121 3456789) 3 Journal of Comparative Physiology B (2018) 188:437–449 438 (Lushchak and Bagnyukova 2006; Lushchak 2011). Lipid peroxidation appears to be one of the most rapid responses to hyperoxia and this stress signal may trigger antioxidant systems (Lushchak and Bagnyukova 2006). In the air- breathing fish Heteropneustes fossilis, 3 h of aerial exposure resulted in an increase in protein carbonylation, an increase in TBARS, and after 6 h, H2O2 concentration was elevated in brain tissue (Paital 2013). In muscle tissue, TBARS and H2O2 were elevated after 3 h of exposure and protein car- bonylation after 6 h (Paital 2014). These results support the conclusion that air-exposed epithelia in fish face oxidative stress, and this does not only affect the air-exposed epithelia, it affects the whole organism. Hypoxic and hyperoxic incubations in jeju and traira Fish were transferred to individual 30-L plastic tanks the day before the experiment. Water was aerated using air stones. Fish were transferred to individual 30-L plastic tanks the day before the experiment. Water was aerated using air stones. The next morning, 75% of the water was siphoned out of the tank and replaced with fresh water. For hypoxic condi- tions, water oxygen was lowered to 1–1.5 mg O2 L− 1 (cor- responding to a PO2 of 2.6–3.9 kPa; hypoxia) by bubbling nitrogen into the tank. Fish were incubated under hypoxic conditions for 4 h. Hyperoxic conditions (28–32 mg O2 L− 1, corresponding to a PO2 of 73–83 kPa; incubation time 4 h) were achieved by bubbling oxygen. Control fish were incubated in aerated normoxic water for 4 h. Water oxygen partial pressure was monitored using the WTW portable oxygen electrode for hypoxic and normoxic conditions, and the Radiometer Clark-type oxygen electrode for hyperoxic conditions, as above. Behavioral observations and in vivo measurements of swimbladder oxygen in jeju Jeju with a catheter inserted into the posterior swimbladder were transferred to translucent 2-L plastic tanks, filled with INPA freshwater until about 6 cm below the top of the tank so the fish could breathe air freely. Outside the tank, the catheter was connected to a gas-tight glass syringe so that gas samples from the posterior swimbladder could be taken without disturbing the fish. i Water was aerated using air stones (normoxia = 7–8 mg O2 L− 1, corresponding to a PO2 of 20 kPa). After an accli- mation period of at least 2 h, fish were observed for 30 min by counting every air-breath. Air-breaths could easily be identified because immediately after the fish left the surface, exhaled air bubbles left the opercular cavity via the opercu- lum. Then the water PO2 was lowered to 1–1.5 mg O2 L− 1 (corresponding to a PO2 of 2.6–3.9 kPa or about 17% satu- ration; hypoxia) by bubbling nitrogen into the tank. After the water was equilibrated to the new PO2, air-breathing activity was again monitored for 30 min by counting every air breath. At the end of this hypoxia treatment, water was aerated and the fish were allowed a recovery period of 2 h. Following this recovery period, water was bubbled with pure oxygen to achieve hyperoxic conditions (28–32 mg O2 L− 1, corresponding to a PO2 of 73–83 kPa or about 450% satura- tion). Air-breathing activity was again counted for 30 min. Water oxygen levels were recorded with a portable oxygen electrode and meter (WTW Multi3410 meter, Weilheim, Germany) for normoxic and hypoxic conditions. Hyperoxic conditions were verified using a Clark-type oxygen electrode and the appropriate meter (Radiometer, E5047 electrode, connected to a PHM 73 m, Copenhagen, Denmark). Surgical procedure in jeju Jeju were anaesthetized with neutralized MS222 (0.1 g L− 1). The body wall was opened close to the lateral line below the end of the dorsal fin. A Clay-Adams PE50 catheter (Becton, 3 Journal of Comparative Physiology B (2018) 188:437–449 439 Dickinson and Co., Franklin Lakes, NJ, USA), slightly flared at the insertion end, was inserted into the caudal end of the posterior swimbladder and anchored in place with a surgical suture and a drop of cyanoacrylate tissue glue (Vetbond™, 3M Co., Maplewood, MN, USA). The catheter was guided through the muscle tissue and the skin using a PE160 sleeve, which was anchored to the PE50 with the glue, then sutured to the body surface. After surgery, fish were transferred to 25-L plastic tanks with aerated INPA freshwater for a recov- ery period of at least 16 h. containing the gas sample was penetrated and the oxygen probe advanced to measure the PO2 of the gas sample. After the measurement, the gas sample was returned to the pos- terior swimbladder. The probe was calibrated with air and nitrogen in the gas phase. Aerial oxygen was set to 100% (20 kPa), and nitrogen was set to zero oxygen. Statistics Data are expressed as mean ± 1 SEM with N giving the num- ber of animals analyzed in each species. GSSG + GSH con- centrations are given as nmol g−1wwt (wet weight), enzyme activities as U mg−1protein (µmol min− 1 mg− 1protein) or as mU mg− 1protein (nmol min− 1 mg− 1protein). For statistical analysis three-way repeated measures ANOVA, followed by the Holm-Sidak Multiple Comparison procedures, was used. Fish species (jeju, traira), tissue (anterior bladder, posterior bladder) and incubation PO2 (normoxia, hypoxia, hyperoxia) were used as parameters (factors) 1, 2 and 3, and enzyme activity or GSH + GSSG concentration as variables (data). PO2 values of the anterior and posterior swimbladder of jeju and traira were analyzed using three-way ANOVA followed by the Bonferoni multiple comparison procedures. PO2 val- ues of the anterior and posterior swimbladder of catheter- ized jeju were analyzed using one-way ANOVA followed by the Holm-Sidak multiple comparison test. In rare cases, the normality test or equal variance test failed using the original data. In this case the data were log transformed prior to sta- tistical analysis. The statistical analysis was performed using SigmaPlot 12.0. Trend/regression lines describing the rela- tion between PO2 and number of air breaths were calculated using Excel using a linear and a logarithmic fit. Significant differences between values were accepted for p < 0.05. Tissue preparation in jeju and traira Fish maintained for 4 h under normoxic, hypoxic or hyper- oxic conditions were rapidly anesthetized with an overdose of neutralized tricaine methanesulfonate (MS222; 0.5 g L− 1) and killed by a sharp blow on the head. To prevent further blood flow to the swimbladder and possible modification of swimbladder gas, fish were then immediately injected with 1 mL of 100 mM KCl into the caudal vein. Preliminary tests showed that this stopped the heart within 30 s. Fish were opened laterally and a small portion of the swimblad- der was carefully exposed for measurement of PO2 of the swimbladder using a PreSens oxygen probe housed in a 22G needle, as above. After penetrating the posterior section of the swimbladder, the probe was carefully advanced into the swimbladder lumen and the PO2 recorded. Subsequently, the oxygen tension of the anterior section of the swimblad- der was recorded, using a separate penetration. The whole procedure starting with anesthesia and recording the oxygen tension of the posterior bladder took between 2 and 3 min, and within less than 4 min recording of swimbladder oxygen tensions was completed. Swimbladder gas samples of 0.2–0.3 mL were taken at the end of the control period, and at the end of the hypoxic or hyperoxic incubation. After disconnecting the syringe from the catheter, the syringe was immediately sealed with parafilm and the PO2 of the gas sample was measured using a PreSens optical oxygen probe (PreSens Precision Sensing, Regensburg, Germany), inserted into a 22G needle. With this needle, the parafilm blocking the tip of the glass syringe After recording gas tensions, the swimbladder was quickly dissected. Connective tissue was removed and the remaining tissue was carefully rinsed with saline, cleaned and blotted dry. The anterior swimbladder tissue of jeju and traira and also the posterior part of traira swimbladder was dissected into small portions and immediately frozen 1 1 3 Journal of Comparative Physiology B (2018) 188:437–449 440 in liquid nitrogen. For jeju, only the vascularized (anterior) section of the posterior swimbladder tissue was used for tis- sue preparation. Tissues were then stored in a biofreezer at − 80 °C until further analysis. Biochemical analyses For determination of total glutathione (GSSG + GSH) con- tent of the frozen tissue samples, tissue extracts were pre- pared using 5% metaphosphoric acid (MPA). The frozen tissues were ground to a fine powder and dissolved 1:5 w/v in 5% MPA. Under ice cooling, the solution was homog- enized using a motorized homogenizer. Extracts were cen- trifuged at 13,000 rpm for 15 min at 4 °C and the super- natant was diluted using assay buffer of the GSSG + GSH Assay kit (STA-312; Cell Biolabs, Inc San Diego, USA). GSSG + GSH concentration was determined using the OxiSelect Total Glutathione (GSSG + GSH) Assay Kit (STA-312) Cell Biolabs, Inc San Diego, USA, following the manufacturer’s instructions. For measurement of enzyme activities, the frozen tissue samples were homogenized on ice in 1:5 w/v of ice-cold homogenization buffer (10 mM of TRIS/HCl, 0.1 mM of disodium EDTA, 150 mM of NaCl, pH 7.5 at 25 °C). Under ice cooling, the tissue was homogenized using a motorized homogenizer. Homogenates were centrifuged at 13,000 rpm for 15 min at 4 °C and appropriate dilutions of the superna- tant were used for the enzyme and protein assays. Enzyme activities were measured using a SpectraMax 384Plus microplate spectrophotometer (Molecular Devices, Sunnyvale, CA, USA) with temperature control at 25 ± 0.1 °C. Glutathione reductase (GR; EC 1.6.4.2.) and glutathione peroxidase (GPx; EC 1.11.1.9.) activities were measured using the Glutathione Reductase Assay Kit (No 703202; Cayman Chemical Company, Ann Harbor, USA), and the Glutathione Peroxidase Assay Kit (No 703102; Cay- man Chemical Company, Ann Harbor, USA). Results 2 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju under normoxic, hypoxic and hyperoxic condi- tions. Different small letters denote significant differences between treatments (p < 0.05); N is given in brackets below the columns Fig. 2 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju under normoxic, hypoxic and hyperoxic condi- tions. Different small letters denote significant differences between treatments (p < 0.05); N is given in brackets below the columns Fig. 2 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju under normoxic, hypoxic and hyperoxic condi- tions. Different small letters denote significant differences between treatments (p < 0.05); N is given in brackets below the columns two species revealed significantly higher levels of oxygen in jeju under both hyperoxic and hypoxic conditions (Fig. 1). Oxygen partial pressure in the posterior bladder of jeju measured on live, freely swimming animals via a cath- eter chronically inserted at the caudal end of the blad- der revealed significantly higher PO2 values (Fig. 2) as compared to the values obtained by puncturing the swim- bladder wall and using the optical fiber (Fig. 1a). Under control conditions, the oxygen partial pressure amounted to 12.34 ± 1.2 kPa (N = 16). A similar value was recorded after 30 min of hypoxia, but after 30 min of hyperoxia PO2 was significantly elevated to 18.1 ± 1.56 kPa (N = 10; Fig. 2). This is similar to the trend seen in the terminally sampled jeju after 4 h of incubation under hyperoxia (Fig. 1a), where the PO2 was also elevated as compared to normoxia. As also observed in the animals incubated for 4 h under hypoxic or hyperoxic conditions, hypoxia stimu- lated air-breathing activity, but under hyperoxic conditions no air-breathing activity was observed. two species revealed significantly higher levels of oxygen in jeju under both hyperoxic and hypoxic conditions (Fig. 1). j j yp yp ( g ) Oxygen partial pressure in the posterior bladder of jeju measured on live, freely swimming animals via a cath- eter chronically inserted at the caudal end of the blad- der revealed significantly higher PO2 values (Fig. 2) as compared to the values obtained by puncturing the swim- bladder wall and using the optical fiber (Fig. 1a). Under control conditions, the oxygen partial pressure amounted to 12.34 ± 1.2 kPa (N = 16). Results The mean body mass of jeju used for the experiments was 95.6 ± 5.9 g (mean ± SEM), and the average fork length was 19.6 ± 0.4 cm (N = 26). The mean body mass of traira amounted to 344.1 ± 21.9 g (mean ± SEM), and the average fork length was 31.4 ± 0.7 cm (N = 22). For both species, body mass and fork length of the individual experimental groups were tested separately and there was no difference in body mass or fork length between control, hypoxic or hyperoxic animals. Catalase (Cat; EC 1.11.1.6.) activity was assayed using the Amplex Red Catalase Assay Kit (A22180; Molecular Probes, Eugene, USA). Superoxide dismutase (SOD; EC 1.15.1.1.) activity was measured following a procedure described by McCord and Fridovich (McCord and Fridovich 1969). Briefly, reactive oxygen species generated from xan- thine in the xanthine oxidase reaction cause a reduction of cytochrome c, which is inhibited by the presence of SOD. One unit of SOD activity is defined as the amount of enzyme (per milligram of protein) that inhibits the reduction of cytochrome c observed in the blank without SOD by 50%. Under normoxia, direct measurement of PO2 in the swimbladder on terminally sampled jeju, by direct pen- etration of the bladder with the oxygen probe, revealed a value of 7.08 ± 2.9 kPa in the anterior bladder, and only 2.6 ± 0.54 kPa (N = 6) in the posterior bladder (Fig. 1a). In traira under normoxia, a PO2 of 4.36 ± 2.24 kPa was detected in the anterior bladder, and 2.42 ± 0.62 kPa (N = 6) in the posterior bladder by the same technique (Fig. 1b). For both species, there was no significant difference between the two parts of the swimbladder. Protein concentration in the homogenate was measured with Coomassie Brilliant Blue G-250 (Bradford 1976) using bovine serum albumin as a standard. In jeju, while 4 h of hypoxic incubation did not cause any change in anterior and posterior swimbladder PO2, 4 h of hyperoxia resulted in a significantly elevated PO2 in the 1 3 Journal of Comparative Physiology B (2018) 188:437–449 441 p y gy ( ) two species revealed significantly higher levels of oxygen in jeju under both hyperoxic and hypoxic conditions (Fig. 1). Results Oxygen partial pressure in the posterior bladder of jeju measured on live, freely swimming animals via a cath- eter chronically inserted at the caudal end of the blad- der revealed significantly higher PO2 values (Fig. 2) as compared to the values obtained by puncturing the swim- bladder wall and using the optical fiber (Fig. 1a). Under control conditions, the oxygen partial pressure amounted to 12.34 ± 1.2 kPa (N = 16). A similar value was recorded after 30 min of hypoxia, but after 30 min of hyperoxia PO2 was significantly elevated to 18.1 ± 1.56 kPa (N = 10; Fig. 2). This is similar to the trend seen in the terminally sampled jeju after 4 h of incubation under hyperoxia (Fig. 1a), where the PO2 was also elevated as compared to normoxia. As also observed in the animals incubated for 4 h under hypoxic or hyperoxic conditions, hypoxia stimu- lated air-breathing activity, but under hyperoxic conditions Fig. 1 Oxygen content in kPA measured by direct puncture in the anterior and posterior swimbladder of jeju (a) and traira (b) under normoxic, hypoxic and hyperoxic conditions. Different capital letters denote significant differences between species, small letters denote significant differences within a species (p < 0.05), bars without let- ters are not significantly different; N is given in brackets below the columns. There was a significant overall difference between the two species Fig. 2 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju under normoxic, hypoxic and hyperoxic condi- tions. Different small letters denote significant differences between treatments (p < 0.05); N is given in brackets below the columns Fig. 1 Oxygen content in kPA measured by direct puncture in the anterior and posterior swimbladder of jeju (a) and traira (b) under normoxic, hypoxic and hyperoxic conditions. Different capital letters denote significant differences between species, small letters denote significant differences within a species (p < 0.05), bars without let- ters are not significantly different; N is given in brackets below the columns. There was a significant overall difference between the two species Fig. 2 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju under normoxic, hypoxic and hyperoxic condi- tions. Different small letters denote significant differences between treatments (p < 0.05); N is given in brackets below the columns Fig. ( f = 0.5699ln(x) + 11.764 (R2 = 0.3642; f = PO2 of the bladder; x = number of breaths taken). Results Jeju: control, N = 6, hyper- oxia, N = 9, hypoxia, N = 8; traira: control, N = 15, hyperoxia, N = 9, hypoxia, N = 9 Fig. 3 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju in relation to the number of breaths taken within an observation period of 30 min in a normoxic and b hypoxic jeju. Under hyperoxic conditions, no air-breathing activity was observed Thus, PO2 initially increased with increasing air-breath- ing activity and the lowest PO2 was detected in fish that did not breathe air during the observation period. Between 5 and 25 breaths per 30 min, however, almost no further increase in the PO2 was detectable. Fig. 4 Catalase activity in U mg− 1 protein in anterior (a) and poste- rior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant dif- ferences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differ- ences between the two species, different small letters denote signifi- cant differences between treatments within a species, bars without letters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyper- oxia, N = 9, hypoxia, N = 8; traira: control, N = 15, hyperoxia, N = 9, hypoxia, N = 9 Under hypoxic conditions, the scatter of the data was reduced, and the PO2 of the bladder decreased in a linear fashion with increasing number of breaths taken (Fig. 3b), as described by the equation: (2 f = −0.4374x + 17.072 (R2 = 0.3173; f = PO2 of the bladder; x = number of breaths taken). (2) The number of breaths taken under hypoxic condi- tions (11.7 ± 1.1 breaths/30 min; N = 13) was signifi- cantly higher than under normoxic conditions (7.9 ± 1.8 breaths/30 min; N = 16). 4 h of hyperoxia. In jeju, catalase activity in the posterior section of the swimbladder was significantly higher than in the anterior section under all three conditions tested, but there was no significant difference between normoxic, hypoxic- or hyperoxic-incubated fish (Fig. 4a). Results A similar value was recorded after 30 min of hypoxia, but after 30 min of hyperoxia PO2 was significantly elevated to 18.1 ± 1.56 kPa (N = 10; Fig. 2). This is similar to the trend seen in the terminally sampled jeju after 4 h of incubation under hyperoxia (Fig. 1a), where the PO2 was also elevated as compared to normoxia. As also observed in the animals incubated for 4 h under hypoxic or hyperoxic conditions, hypoxia stimu- lated air-breathing activity, but under hyperoxic conditions no air-breathing activity was observed. Fig. 1 Oxygen content in kPA measured by direct puncture in the anterior and posterior swimbladder of jeju (a) and traira (b) under normoxic, hypoxic and hyperoxic conditions. Different capital letters denote significant differences between species, small letters denote significant differences within a species (p < 0.05), bars without let- ters are not significantly different; N is given in brackets below the columns. There was a significant overall difference between the two species posterior bladder with 11.08 ± 1.8 kPa (N = 5; Fig. 1a). PO2 in the anterior bladder was also elevated to approximately the same value, but the difference was not yet significant. In Fig. 3a, the oxygen partial pressure of the posterior swimbladder of jeju under normoxia, as sampled by cath- eter in freely swimming animals, was plotted against the number of air-breaths taken in the preceding 30 min. The data showed a remarkable variability, but the overall rela- tionship appeared to be hyperbolic and was well described by the equation: fi Visual observation of the swimming activity of traira revealed almost no surfacing activity under any of the experimental treatments. Fish were quiet on the bottom of the tank, and even ventilatory activity was hardly detectable under hypoxic conditions. f = 0.5699ln(x) + 11.764 (R2 = 0.3642; f = PO2 of the bladder; x = number of breaths taken). In traira neither 4 h of hypoxia (1–1.5 mg O2 L− 1) nor 4 h of hyperoxia (28–32 mg O2 L− 1) caused any significant change in the PO2 of the anterior and posterior swimbladder (Fig. 1b). 1 3 Results A comparison of swimbladder PO2 between the (1) 1 3 Journal of Comparative Physiology B (2018) 188:437–449 442 442 Journal of Comparative Physiology B (2018) 188:437 449 Thus, PO2 initially increased with increasing air-breath- ing activity and the lowest PO2 was detected in fish that did not breathe air during the observation period. Between 5 and 25 breaths per 30 min, however, almost no further increase in the PO2 was detectable. Under hypoxic conditions, the scatter of the data was reduced, and the PO2 of the bladder decreased in a linear fashion with increasing number of breaths taken (Fig. 3b), as described by the equation: The number of breaths taken under hypoxic condi- (2) f = −0.4374x + 17.072 (R2 = 0.3173; f = PO2 of the bladder; x = number of breaths taken). 4 h of hyperoxia. In jeju, catalase activity in the posterior Fig. 3 Oxygen content in kPa measured via a catheter in the posterior swimbladder of jeju in relation to the number of breaths taken within an observation period of 30 min in a normoxic and b hypoxic jeju. Under hyperoxic conditions, no air-breathing activity was observed Fig. 4 Catalase activity in U mg− 1 protein in anterior (a) and poste- rior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant dif- ferences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differ- ences between the two species, different small letters denote signifi- cant differences between treatments within a species, bars without letters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyper- oxia, N = 9, hypoxia, N = 8; traira: control, N = 15, hyperoxia, N = 9, hypoxia, N = 9 Fig. 4 Catalase activity in U mg− 1 protein in anterior (a) and poste- rior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant dif- ferences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differ- ences between the two species, different small letters denote signifi- cant differences between treatments within a species, bars without letters are not significantly different. There was also a significant overall difference between the two species. Results 6 Glutathione reductase (GR) activity in mU mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small letters denote significant differences between treatments within a species, bars without letters are not significantly different There was also a Fig. 5 Superoxide dismutase (SOD) activity in U mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small let- ters denote significant differences between treatments within a spe- cies, bars without letters are not significantly different. Jeju: control, N = 11, hyperoxia, N = 8, hypoxia, N = 8; traira: control, N = 14, hyperoxia, N = 9, hypoxia, N = 9 Fig. 6 Glutathione reductase (GR) activity in mU mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small letters denote significant differences between treatments within a species, bars without letters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyperoxia, N = 8, hypoxia, N = 8; traira: control, N = 14, hyper- oxia, N = 9, hypoxia, N = 8 50% in both parts of the swimbladder as compared to nor- moxic- or hyperoxic-incubated fish (Fig. 4b). The overall comparison revealed significant differences between the two species, with specific significant differences in nor- moxia and hyperoxia, but not in hypoxia. Catalase activity in the posterior section of traira was significantly different from the activity measured in the posterior section of jeju. hyperoxic incubation (Fig. 5a). In traira, however, hyper- oxic conditions caused a 1.6- to two-fold increase in SOD activity in the anterior and posterior swimbladder, respec- tively, and this was significantly different from control values as well as from hypoxic values (Fig. Results In traira, no difference was detected between the anterior and the posterior part of the swimbladder, but following 4 h of hypoxia-catalase activity was significantly lower by about As a next step, we measured the activity of ROS- degrading enzymes in anterior and posterior swimbladder tissue of both species following incubation under nor- moxic control conditions, after 4 h of hypoxia and after 1 3 Journal of Comparative Physiology B (2018) 188:437–449 443 50% in both parts of the swimbladder as compared to nor- Fig. 5 Superoxide dismutase (SOD) activity in U mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small let- ters denote significant differences between treatments within a spe- cies, bars without letters are not significantly different. Jeju: control, N = 11, hyperoxia, N = 8, hypoxia, N = 8; traira: control, N = 14, hyperoxia, N = 9, hypoxia, N = 9 Fig. 6 Glutathione reductase (GR) activity in mU mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small letters denote significant differences between treatments within a species, bars without letters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyperoxia, N = 8, hypoxia, N = 8; traira: control, N = 14, hyper- oxia, N = 9, hypoxia, N = 8 Fig. 5 Superoxide dismutase (SOD) activity in U mg− 1 protein in anterior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small let- ters denote significant differences between treatments within a spe- cies, bars without letters are not significantly different. Jeju: control, Fig. Results In traira, GPx activities in the anterior and pos- terior section of the swimbladder were always similar to Fig. 7 Glutathione peroxidase activity in mU mg− 1 protein in ante- rior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Note the, differ- ent scales in panels a and b. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small letters denote significant dif- ferences between treatments within a species, bars without letters are not significantly different. There was also a significant overall differ- ence between the two species. Jeju: control, N = 10, hyperoxia, N = 9, hypoxia, N = 5; traira: control, N = 11, hyperoxia, N = 9, hypoxia, N = 6 Fig. 8 Total GSSG + GSH concentration in anterior (a) and poste- rior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant dif- ferences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differ- ences between the two species, different small letters denote signifi- cant differences between treatments within a species, bars without let- ters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyperoxia, N = 7, hypoxia, N = 8; traira: control, N = 6, hyperoxia, N = 3, hypoxia, N = 6 Fig. 7 Glutathione peroxidase activity in mU mg− 1 protein in ante- rior (a) and posterior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Note the, differ- ent scales in panels a and b. Asterisks mark significant differences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differences between the two species, different small letters denote significant dif- ferences between treatments within a species, bars without letters are not significantly different. There was also a significant overall differ- ence between the two species. Jeju: control, N = 10, hyperoxia, N = 9, hypoxia, N = 5; traira: control, N = 11, hyperoxia, N = 9, hypoxia, N = 6 Fig. Results 5b). Under hypoxic conditions, SOD activity in the posterior bladder was significantly lower than in the anterior bladder. The comparison between the two species showed that under normoxic control conditions, SOD activity in jeju was sig- nificantly higher than in traira. However, under hyperoxic conditions, SOD activity was significantly higher in traira, as compared to jeju. hyperoxic incubation (Fig. 5a). In traira, however, hyper- oxic conditions caused a 1.6- to two-fold increase in SOD activity in the anterior and posterior swimbladder, respec- tively, and this was significantly different from control values as well as from hypoxic values (Fig. 5b). Under hypoxic conditions, SOD activity in the posterior bladder was significantly lower than in the anterior bladder. The comparison between the two species showed that under normoxic control conditions, SOD activity in jeju was sig- nificantly higher than in traira. However, under hyperoxic conditions, SOD activity was significantly higher in traira, as compared to jeju. Superoxide dismutase activity measured in jeju swim- bladder tissue was similar in both sections of the swim- bladder under normoxic and hypoxic conditions, but under hyperoxia SOD activity in the posterior bladder was higher than in the anterior bladder (Fig. 5a). In jeju, overall no differences could be detected in response to hypoxic or 1 3 Journal of Comparative Physiology B (2018) 188:437–449 444 Measurement of GR activity in jeju swimbladder revealed similar activity in both sections of the swimbladder, and no differences could be detected in response to hypoxic or hyperoxic incubation (Fig. 6a). In traira anterior and pos- terior swimbladder tissue, GR activity was also quite simi- lar, but again, following hyperoxic incubation, the activity in both sections of traira swimbladder was significantly higher than under normoxia or following hypoxic incuba- tion (Fig. 6b). In hyperoxia, GR activity was as almost twice as high as in normoxia. The comparison of jeju and traira showed overall significant difference between the two spe- cies. Specifically, there was significantly higher GR activity in jeju swimbladder tissue under normoxic conditions and under hypoxia, but no difference was detected under hyper- oxic conditions. Under normoxia, GPx activity was similar in the anterior and posterior section of jeju swimbladder (Fig. 7a). Hypoxic as well as hyperoxic incubation resulted in a significant enhancement of GPx activity in the posterior part of jeju swimbladder. Glutathione peroxidase activity increased 2.8- fold under hyperoxic conditions and 3.4-fold under hypoxic conditions. Results 8 Total GSSG + GSH concentration in anterior (a) and poste- rior (p) swimbladder tissue of jeju (a) and traira (b) under normoxic, hyperoxic and hypoxic conditions. Asterisks mark significant dif- ferences (p < 0.05) between posterior and anterior values within the same species and treatment. Capital letters denote significant differ- ences between the two species, different small letters denote signifi- cant differences between treatments within a species, bars without let- ters are not significantly different. There was also a significant overall difference between the two species. Jeju: control, N = 6, hyperoxia, N = 7, hypoxia, N = 8; traira: control, N = 6, hyperoxia, N = 3, hypoxia, N = 6 1 3 445 Journal of Comparative Physiology B (2018) 188:437–449 Therefore, a 100 g of jeju would consume about 19 µmol of oxygen within 2 min. A 100 g of jeju would have a swim- bladder volume of about 5 mL (Kramer 1978). Assuming an oxygen content of 60% (12 kPa) of aerial oxygen (meas- ured in the catheter experiments), this would give an oxygen content of about 0.6 mL, equivalent to 26 µmol at STPD. Accordingly, given the time needed for anaesthesia and the further up to 30 s for the time needed for the injection of KCl to stop the heart (1.5–2 min total), the continuation of blood flow could indeed have appreciably reduced the oxygen con- tent of the posterior swimbladder. each other, but varied according to the treatment (Fig. 7b). Under hyperoxia, GPx activity was significantly elevated as compared to hypoxia, but it was not different from the activity measured under control conditions. Overall, GPx activity was significantly higher in jeju as compared to traira, though a specific significant difference was seen only in the hypoxia treatment. Measurement of total glutathione concentration (GSH + GSSG) again resulted in similar concentrations in the anterior and posterior section of jeju swimbladder. Comparing thevalues obtained under normoxic, hyperoxic and hypoxic conditions, however, showed a significant ele- vation of total glutathione concentration under hypoxic as well as under hyperoxic conditions (Fig. 8a). In traira, similar glutathione concentrations were detected under all conditions in both sections of the swimbladder. In the anterior section, the concentration always appeared to be higher, and this difference was significant for control and for hypoxic fish. Comparing the jeju with traira revealed significantly higher concentrations of total glutathione in jeju under all conditions. Critique of methods Swimbladder puncture with a 22G needle and a fiber optic system has been used to measure oxygen content in the anterior and posterior swimbladder in quickly sacrificed animals, and chronic catheterization by inserting PE50 tub- ing into the caudal end of the posterior bladder in freely swimming animals. The results obtained with the catheter were significantly higher and consistent with values reported by Kramer (Kramer 1978), who collected gas by punctur- ing the posterior tip of the bladder and collecting gas in a water-filled funnel, and by Farrell and Randall (Farrell and Randall 1978), who cannulated the posterior bladder simi- lar to our procedure. Comparing the changes of jeju poste- rior bladder oxygen content under normoxic, hypoxic and hyperoxic conditions, both procedures gave similar results qualitatively. Incubation under hypoxic conditions did not change posterior bladder oxygen content, but under hyper- oxia, oxygen content increased. Both procedures appeared to have intrinsic limitations. Using the direct puncture method, the fish had to be anesthetized, and the swimbladder had to be exposed by dissection before the swimbladder wall could be penetrated with the 22G needle. Although the whole pro- cedure was completed within 2 or 3 min, oxygen could have been removed from the bladder during this time and not replaced, resulting in lower oxygen values. In a previous study, we measured an oxygen uptake of 5.6 ± 0.6 µmol g− 1 h− 1 for jeju at a temperature of 29–32 °C (Wood et al. 2016). Results A problem of the catheter in turn is the tightness, and a tiny leak in the gas phase or diffusion through the PE tub- ing may have caused an increase in the oxygen content. The swimbladder typically contains some moisture to avoid desiccation of the epithelia, which sometimes hinders the collection of a clean gas sample. The dead space of the cath- eter may be equilibrated with a mixture of atmospheric and swimbladder gas, which also may result in elevated gas con- tents. Thus, while the direct penetration method may result in an underestimation of swimbladder oxygen content, the catheter may result in an overestimation. We, therefore, assume the real value to be in between these two results. Convincingly, however, both measurements consistently showed that jeju-posterior swimbladder oxygen content was not modified under hypoxic conditions, but it significantly increased under hyperoxic conditions. Flexibility of the ROS defense system The comparison of the ROS defense capacity of the jeju and the traira confirmed our previous study revealing a better ROS defense capacity in the facultative air-breathing jeju (Pelster et al. 2016), although the differences were not as clear as in the previous study. In the present study, catalase activity was even slightly higher in the traira as compared to jeju. Nevertheless, SOD and GR activity and also the concentration of GSH + GSSG were higher in jeju under control conditions. In addition, GR and GPx activity and GSH + GSSG concentration overall were higher in jeju as compared to traira. This confirms our previous observation that the glutathione-based ROS defense system appears more important in jeju than immediate ROS breakdown by SOD (Pelster et al. 2016). The differences in the responses in comparison to our previous study most likely appear to be attributable to the lower temperature used for the present experiments. ROS formation is temperature dependent and increases with higher temperatures. Therefore, the variable oxygen tensions encountered with an enhanced air-breath- ing activity may stimulate the expression of a well-organ- ized ROS defense system in the jeju, especially at higher temperature.l In traira under hyperoxic conditions, swimbladder oxygen content was not different from control or hypoxia-incubated animals, supporting the notion that the swimbladder is not used for gas exchange and not intensely perfused. Accord- ingly, changes in arterial PO2 caused by changing water PO2 will only in the long run influence swimbladder PO2. In the jeju, however, oxygen content of the posterior bladder increased significantly, as observed in both directly punc- tured swimbladders as well as in chronically catheterized swimbladders. No air-breathing was observed under hyper- oxia in our experiments, therefore, the oxygen must have been taken up via the gills and transferred to other organs, including the posterior swimbladder. The posterior bladder has a rich vascularization and thus blood supply, whereas the anterior swimbladder is poorly vascularized. This together with the higher PO2 value recorded in the anterior bladder under normoxic conditions explain why the oxygen content in the anterior bladder was not yet significantly elevated under hyperoxic conditions. In contrast to our previous study, where air breathing con- tinued during hyperoxia (Pelster et al. 2016), in the present study, no air-breathing activity was observed in jeju under hyperoxic conditions. Swimbladder function under hypoxia and hyperoxia As expected, the swimbladder of the jeju had a significantly higher oxygen content than the swimbladder of traira. Gases are resorbed from non-ventilated cavities according to their physical solubility, resulting in an accumulation of the less soluble gases (Piiper et al. 1962; Piiper 1965). The swim- bladder of traira is used as a buoyancy structure and hardly ventilated (Rantin et al. 1992; Val and Almeida-Val 1995; Glass and Rantin 2009; Pelster et al. 2016), therefore, the less soluble nitrogen should accumulate in this bladder. The jeju swimbladder, in turn, is frequently ventilated, and the inhaled gas initially enters the anterior section of the swim- bladder (Randall et al. 1981). The posterior part of the jeju swimbladder is filled from the anterior part. It is highly vas- cularized and used as a gas exchange organ, and we meas- ured a lower PO2 in posterior bladder as compared to the anterior part, although this difference was not yet significant. fi Under normoxic conditions a large scatter in the oxygen content of the swimbladder was observed, which in part was attributable to the time difference between the last breath and sampling of swimbladder gas. In line with the results obtained for traira, the lowest oxygen content in jeju under normoxia was detected in swimbladders that were hardly ventilated. Between 5 and 25 breaths per 30 min, however, 1 3 Journal of Comparative Physiology B (2018) 188:437–449 446 have been performed at temperatures between 25 and 28 °C. The higher temperature significantly stimulates metabolic activity and oxygen requirements, but the solubility of oxy- gen in water decreases with temperature, impairing aquatic oxygen uptake via the gills. It is, therefore, conceivable that at high temperatures jeju resort to aerial respiration even under hyperoxic conditions to supplement oxygen uptake. oxygen content was quite variable and no clear correlation with the number of breaths taken was detectable. In addition to ventilation the intensity with which the bladder was used as an oxygen resource must have an influence, and removal of oxygen from the bladder is dependent on blood flow. In an intensely ventilated swimbladder with frequent gas exchange between the anterior and the posterior bladder, the differ- ences in oxygen content between the two parts is expected to become smaller. Interestingly, under severe hypoxia in traira, almost no activity was detectable, and even ventilatory activity was often hard to detect. Swimbladder function under hypoxia and hyperoxia This again contrasts with our previous study performed at a higher temperature, where traira started to perform aquatic surface respiration during severe hypoxia (Pelster et al. 2016). The present observations suggest that at 26 ± 1 °C, traira adopted a severe depression of metabolic activity to withstand the hypoxic period. Occasionally, this behavior was also seen in jeju, suggesting that the jeju has two options to survive under hypoxia, either resorting to intensive air breathing, or reducing activity and referring to metabolic depression, as does the traira. Under hypoxic conditions, ventilatory activity of the jeju increased, as reported also in previous studies (Kramer 1978; Stevens and Holeton 1978; Oliveira et al. 2004; Perry et al. 2004; Juca-Chagas 2004; Pelster et al. 2016), but the oxygen content did not increase. Indeed, a negative correla- tion between posterior bladder oxygen content and breathing frequency was observed. Testing the PO2 in the gas space above the hypoxic water revealed that in this set up, PO2 in the gas space may drop to 15–20% below aerial PO2 (i.e. to 16–17 kPa). Thus, air-breathing jeju would pick up slightly hypoxic air, but nevertheless the PO2 of this air was much higher than water PO2 with a value of only 2.6–3.9 kPa. The constant PO2 in the swimbladder, therefore, suggests that the intensely ventilated swimbladder is indeed used to supply oxygen to other tissues. 3 Flexibility of the ROS defense system Analysis of the hypoxia response of epaulette shark, threespine stickleback and rainbow trout combined with a literature survey looking at 13 species of fish and one shark revealed that fish in general do not show an increase in antioxidant defense response under hypoxia, and if a response is seen, it is not related to the hypoxia tolerance of the species (Leveelahti et al. 2014). Following reoxygenation after a hypoxic episode, often an enhancement of the ROS defense capacity is observed, which is discussed as a prepa- ration for the oxidative stress encountered during recovery from hypoxia (Hermes-Lima et al. 1998; Lushchak and Bag- nyukova 2006; Welker et al. 2013).if For the jeju variable oxygen content in the bladder and thus in blood are frequently expected, depending on the air-breathing activity, and we indeed observed a significant scatter in the oxygen content of the swimbladder under nor- moxic conditions. Accordingly, the ROS defense system is probably pre-adjusted to variable oxygen concentrations. This may explain why in our experiments we did not observe a marked response in the ROS-detoxifying systems of swim- bladders in animals exposed to either hypoxia or hyperoxia; only GPx activity was elevated in hyperoxic animals. In line with the increase in GPx activity during hyperoxia, total glutathione concentration was elevated in hyperoxia. The present study revealed significant differences in phys- iology and performance of the swimbladder of two eryth- rinid fish. The use of the swimbladder as an air-breathing organ in the jeju results in highly variable PO2 values in the swimbladder, depending on the frequency of breathing air. Swimbladder PO2 also appeared to be related to blood flow to the swimbladder, as suggested by the rapid increase in PO2 (within 30 min) under hyperoxic conditions when no air breaths were taken, and the high variability of blad- der PO2 at a similar number of breaths taken under nor- moxic conditions. The high variability of swimbladder PO2 coincided with an improved ROS defense system, as compared to swimbladder tissue not used for aerial respi- ration. In consequence, hypoxic or hyperoxic conditions did not have a strong influence on the capacity of the ROS defense system. In the traira, which uses the swimbladder as a buoyancy organ and not for breathing air, swimbladder PO2 values remained constant, irrespective of environmental PO2. Flexibility of the ROS defense system A significant reduction in air-breath- ing frequency under hyperoxic conditions has also been reported by Farrell and Randall (Farrell and Randall 1978). The most likely explanation for this difference appears to be the temperature. Our previous study has been performed on board a ship with temperatures of 30–35 °C (Pelster et al. 2016), while the study of Farrell and Randall and this study Considering the flexibility of the ROS defense system, our results revealed that 4 h of hypoxia or hyperoxia were sufficient to induce changes in some of the ROS-degrad- ing enzymes. The responsiveness observed in traira by far exceeded the responsiveness detected in jeju swimbladder tissue. While in the jeju an elevation of activity was only observed for GPx under hyperoxic and hypoxic condi- tions, in traira, in turn, activities of SOD, GR and GPx were increased during hyperoxia, but catalase was reduced under 3 3 Journal of Comparative Physiology B (2018) 188:437–449 447 hypoxia. This was surprising because our data demonstrated that the traira swimbladder plays no role in gas exchange, and the oxygen content was constant under all experimental conditions tested. Changing oxygen availability is detected by oxygen receptors located in the gills, and transmitted to the central nervous system (Lopes et al. 2010; Zachar and Jonz 2012; Milsom 2012). From there, the stimulus to modify the ROS defense capacity must reach various organs in the whole body; conceivably, the signal could be altered blood O2 or ROS levels, and/or other blood-borne or neural pathways. Previous studies have demonstrated that a modi- fication of the ROS defense system is not only detectable in the gills but also in muscle tissue or in the liver, organs not in direct contact with the environmental water (see Introduc- tion). Thus, the signaling cascade also included the swim- bladder in traira. 2000) but in goldfish, most enzyme activities tested were not changed during hyperoxic exposure of 12 h (Lushchak et al. 2005a). Goldfish exposed to anoxic conditions increased catalase activity in liver, GPx and G6PDH activity in brain (Lush- chak et al. 2001) and hypoxia increased SOD activity in liver, brain and gills in common carp (Vig and Nemcsok 1989), and catalase and GPx activity in brain (Lushchak et al. 2005b). References Lygren B, Hamre K, Waagbo R (2000) Effect of induced hyperoxia on the antioxidant status of Atlantic salmon Salmo salar L. fed three different levels of dietary vitamin E. 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J Exp Zool 232:29–39 Gilmour KM, Perry SF (1994) The effects of hypoxia, hyperoxia or hypercapnia on the acid-base disequilibrium in the arterial blood of rainbow trout. J Exp Biol 192:269–284 i Muusze B, Marcon J, Van den Thillart G, Almeida-Val V (1998) Hypoxia tolerance of Amazon fish: respirometry and energy metabolism of the cichlid Astronotus ocellatus. Comp Biochem Physiol Part A 120:151–156 Glass ML, Rantin FT (2009) Gas exchange and control of respiration in air-breathing Teleost fish. In: Glass ML, Wood SC (eds) Cardio- respiratory control in vertebrates—evolutionary and evolutionary aspects. Springer, Heidelberg, pp 99–120 Oehlers LP, Perez AN, Walter RB (2007) Detection of hypoxia-related proteins in medaka (Oryzias latipes) brain tissue by difference gel electrophoresis and de novo sequencing of 4-sulfophenyl iso- thiocyanate-derivatized peptides by matrix-assisted laser desorp- tion/ionization time-of-flight mass spectrometry. 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Flexibility of the ROS defense system Am J Physiol Regul Integr Comp Physiol 280(1):R100–R107 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecom- mons.org/licenses/by/4.0/), which permits unrestricted use, distribu- tion, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecom- mons.org/licenses/by/4.0/), which permits unrestricted use, distribu- tion, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Lushchak VI, Bagnyukova TV, Husak VV, Luzhna LI, Lushchak OV, Storey KB (2005a) Hyperoxia results in transient oxidative stress and an adaptive response by antioxidant enzymes in goldfish tis- sues. Int J Biochem Cell Biol 37(8):1670–1680 Lushchak VI, Bagnyukova TV, Lushchak OV, Storey JM, Storey KB (2005b) Hypoxia and recovery perturb free radical processes and antioxidant potential in common carp (Cyprinus carpio) tissues. Int J Biochem Cell Biol 37(6):1319–1330 Int J Biochem Cell Biol 37(6):1319–1330f Flexibility of the ROS defense system Changing environmental PO2 values, however, caused a much stronger response in the ROS defense system than observed in jeju. This indicated that changes in environmen- tal PO2 may influence the ROS defense system not only in tissues immediately exposed to the varying PO2 but also in organs not directly confronted with the modified PO2. g yp While our data clearly show responses of the ROS defense system to changing oxygen concentration in the swimblad- der, experimental deflation of the swimbladder of Opsanus tau with subsequent increase in swimbladder oxygen content did not result in a change in enzyme activities of the ROS defense system (Morris and Albright 1984). In the European eel, however, the process of silvering, which is the prepara- tion for the deep seawater spawning migration, is connected to a significant improvement of the ROS defense system (Schneebauer et al. 2016). Very probably, this is because much higher oxygen partial pressures are expected to occur in the swimbladder during exposure to elevated hydrostatic pressure during spawning migration than during freshwater residence (Pelster 2015). The response of the ROS defense system to changing oxygen availability appears to be quite variable and species specific. Juvenile Oncorhynchus mykiss exposed for 4 h to hyperoxic conditions (> 40 mg O2 L− 1) show an elevated catalase activity in gills and liver 24 h after the exposure, but not at 1 to 12 h. SOD, total GPx and GR activity, however, were not affected (Ritola et al. 2002a). A very rapid response was observed in red blood cells, which showed an increase in SOD and catalase activity after only 5 min of exposure to hyperoxia (Ritola et al. 2002b). In liver of Atlantic salmon, 140–150% oxygen saturation for several weeks increased SOD and catalase activities (Lygren et al. Acknowledgements Open access funding provided by University of Innsbruck and Medical University of Innsbruck. Financial support by INCT ADAPTA-CNPq/FAPEAM, Science without Borders (Brazil), and NSERC Discovery (Canada) is gratefully acknowledged. ALV is a recipient of a research fellowship from Brazilian CNPq. EJ was sup- ported UBC Go Global. The authors would like to thank Gilda Pelster for support in performing the biochemical assays, and Drs Bill Milsom and Colin Brauner, UBC, for the loan of the Presens system. 1 3 Journal of Comparative Physiology B (2018) 188:437–449 448 during anoxia and reoxygenation. References J Appl Physiol 17:268–274 Randall DJ, Burggren WW, Farrell AP, Haswell MS (1981) The evolu- tion of air-breathing in vertebrates. Cambridge University Press, Cambridge Lushchak VI, Bagnyukova TV (2007) Hypoxia induces oxidative stress in tissues of a goby, the rotan Perccottus glenii. Comp Biochem Physiol B Biochem Mol Biol 148(4):390–397 Rantin FT, Kalinin AL, Glass ML, Fernandes MN (1992) Respiratory responses to hypoxia in relation to mode of life of two erythrinid Lushchak VI, Lushchak LP, Mota AA, Hermes-Lima M (2001) Oxida- tive stress and antioxidant defenses in goldfish Carassius auratus 1 3 Journal of Comparative Physiology B (2018) 188:437–449 449 species (Hoplias malabaricus and Hoplias lacerdae). J Fish Biol 41(5):805–812 Val AL, Almeida-Val VMF (1995) Fishes of the Amazon and their environment. Springer, Berlinf Ritola O, Livingstone DR, Peters LD, Lindström-Seppä P (2002a) Antioxidant processes are affected in juvenile rainbow trout (Oncorhynchus mykiss) exposed to ozone and oxygen-supersatu- rated water. Aquaculture 210:1–19 Vig E, Nemcsok J (1989) The effects of hypoxia and paraquat on the superoxide dismutase activity in different organs of carp, Cyprinus carpio L. J Fish Biol 35(1):23–25 Welker AF, Moreira DC, Campos EG, Hermes-Lima M (2013) Role of redox metabolism for adaptation of aquatic animals to drastic changes in oxygen availability. Comp Biochem Physiol A Mol Integr Physiol 165(4):384–404 Ritola O, Peters LD, Livingstone DR, Lindström-Seppä P (2002b) Effects of in vitro exposure to ozone and/or hyperoxia on super- oxide dismutase, catalase, glutathione and lipid peroxidation in red blood cells and plasma of rainbow trout, Oncorhynchus mykiss (Walbaum). Aquacult Res 33(3):165–175 g y Wilhelm Filho DW (1996) Fish antioxidant defenses—a comparative approach. Brazilian J Med Biol Res 29:1735–1742 Schneebauer G, Hanel R, Pelster B (2016) Anguillicola crassus impairs the silvering-related enhancements of the ROS defense capacity in swimbladder tissue of the European eel (Anguilla anguilla). J Comp Physiol B 186(7):867–877 Wilhelm Filho D, Torres MA, Zaniboni-Filho E, Pedrosa RC (2005) Effect of different oxygen tensions on weight gain, feed conver- sion, and antioxidant status in piapara., Leporinus elongatus (Valenciennes, 1847). Aquaculture 244:349–357 p y Stevens ED, Holeton GF (1978) The partitioning of oxygen uptake from air and from water by erythrinids. References Can J Zool 56(4):965–969 Wood CM, Pelster B, Giacomin M, Sadauskas-Henrique H, Almeida- Val VM, Val AL (2016) The transition from water-breathing to air-breathing is associated with a shift in ion uptake from gills to gut: a study of two closely related erythrinid teleosts, Hopleryth- rinus unitaeniatus and Hoplias malabaricus. J Comp Physiol B 186(4):431–445 Storey KB (1996) Oxidative stress: animal adaptations in nature. 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https://openalex.org/W1509897417_16	Spanish-Science-Pile	Open Science	Various open science	2,015	Metodología para el diagnóstico de averías en motores de inducción mediante el análisis de corrientes estatóricas transitorias utilizando átomos tiempo-frecuencia.	None	Spanish	Spoken	7,203	12,251	En los ensayos realizados, la excentricidad ha sido generada por medio de un alineamiento inadecuado entre el eje del rotor y el de su carga. Dicho alineamiento inadecuado ha sido causado por la colocación de una pletina de acero en forma de L de 3 milímetros de espesor debajo de uno de los extremos de la plataforma, tal y como se muestra en la fotografía tomada al montaje (Fig. 8.1). Las consecuencias de este tipo de 324 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. desalineamiento entre el motor y la carga en términos de la excentricidad generada han sido estudiados en [103], mostrando que el desalineamiento del eje induce cargas en los rodamientos, las cuales resultan ser suficientemente elevadas para tener un impacto significativo en la excentricidad del entrehierro. Además, los efectos de esta excentricidad creada artificialmente, han sido analizados con profundidad en [9], a través del espectro de la corriente del estator del motor de inducción alimentado mediante el inversor y operando en régimen permanente. Fig. 8. 1 Fotografía del montaje realizado en el laboratorio: freno de polvo magnético, motor de inducción de jaula de ardilla (ambos sujetados con cinchas) y detalle de la cuña posicionada en un extremo para forzar un desalineamiento entre los ejes del motor y de la carga. 8.1.3.- Tipos de transitorios. En primer lugar, el motor de inducción ha sido ensayado desconectado de la carga, sin provocar ningún tipo de fallo artificial, con la intención de detectar cualquier tipo de excentricidad inherente, la cual, tal y como se verá en el siguiente apartado, ha resultado ser prácticamente inexistente. Seguidamente, se ha provocado artificialmente una excentricidad en el motor de inducción, y se ha capturado su corriente de estator durante distintos tipos de transitorios. Tal y como se ha comentado anteriormente, resulta fundamental disponer de una metodología de diagnóstico capaz de analizar motores de inducción trabajando en régimen transitorio. Las aplicaciones en las que se alimentan los motores de inducción mediante un inversor suelen ser de velocidad variable, en las cuales los motores raramente trabajan en régimen permanente, siendo además este régimen demasiado corto en caso de aparecer. La velocidad varía cambiando la frecuencia asignada del inversor mientras la frecuencia de deslizamiento se mantiene en valores muy reducidos. 325 Capítulo 8. Este tipo de transitorio resulta muy común: la frecuencia asignada puede aumentar, como es el caso de un arranque, o puede disminuir para bajar la velocidad. Por supuesto, a este tipo de transitorios se unen los provocados por una oscilación de carga. En definitiva, hay varios tipos de transitorios, debiendo enfrentarse la metodología de diagnóstico a uno u otro dependiendo del tipo de aplicación: varios arranques, aumentos y disminuciones de la frecuencia asignada o variaciones de carga con o sin compensación del deslizamiento. 8.1.4.- Instrumentación de medida utilizada. En definitiva, se han realizado varios ensayos en varias condiciones de funcionamiento, capturando en cada caso la corriente transitoria absorbida por el estator. Todas las corrientes capturadas, las cuales se muestran en la Fig. 8.2, han sido capturadas utilizando un sensor de efecto Hall con una frecuencia de muestreo de 5 kHz. Anteriormente a su análisis utilizando MatLab, la frecuencia de muestreo ha sido reducida a 1 kHz utilizando la función decimate de MatLab, pues el límite superior de frecuencia analizado es 100 Hz, resultando más que suficiente la frecuencia de muestreo de 1 kHz, reduciendo de esta forma el tiempo de computación al trabajar con señales con un menor número de puntos. 8.1.5.- Resumen de corrientes capturadas. El conjunto de las corrientes capturadas se muestran en la siguiente figura: Fig. 8. 2 Corrientes del estator del motor de inducción capturadas durante los cinco ensayos realizados. En primer lugar, tal y como se ha avanzado, se ha ensayado el motor funcionando en régimen permanente desacoplado de la carga, sin crear ningún tipo de 326 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. excentricidad (Fig. 8.2, primera corriente). Las cuatro siguientes corrientes han sido capturadas con el motor acoplado a la carga y generando la excentricidad artificial. Así pues, durante el segundo ensayo se ha programado el inversor para que aumente linealmente la frecuencia asignada desde 0 hasta 50 Hz en aproximadamente 2.5 segundos. Se trata pues de un arranque, siendo la corriente capturada la segunda de la Fig. 2.9. Durante el tercer ensayo, en un segundo se ha disminuido la frecuencia también linealmente desde 50 Hz hasta 33, encontrando tanto antes como después los respectivos regímenes permanentes (Fig. 2.9, tercera corriente). Tanto el cuarto como el quinto ensayo (corrientes cuarta y quinta de la Fig. 2.9) han consistido en una sobrecarga del motor. La única diferencia entre ambos ensayos es que en el quinto la compensación del deslizamiento durante la variación de carga está activada (la frecuencia de la FC varía) y en el cuarto está desactivada (la frecuencia de la FC permanece constante). 327 Capítulo 8. 8.2.- Aplicación de la metodología. En el presente apartado se aplica la metodología de diagnóstico desarrollada en la tesis a las señales capturadas según lo indicado en el apartado anterior, mostrando los resultados obtenidos. A tal efecto se sigue la “Guía de aplicación de la metodología de diagnóstico” presentada en el apartado 4.7, detallándose las particularidades de los análisis realizados. De especial importancia es el apartado 8.2.4, en el que se comenta la elección de las pendientes de las cajas de Heisenberg utilizadas en cada caso. En el siguiente apartado (8.3) se analizan los resultados obtenidos concluyendo el diagnóstico. 8.2.1.- Captura de la corriente. El primer paso para diagnosticar una máquina eléctrica utilizando la metodología desarrollada en la presente tesis es capturar una de sus corrientes. Con lo que respecta al diagnóstico de motores de inducción realizado en el presente capítulo, este primer paso ya ha sido expuesto en el punto anterior al comentar los montajes experimentales y la correspondiente captura de distintas corrientes estatóricas del motor a diagnosticar en distintas condiciones transitorias. 8.2.2.- Estimación de las evoluciones de las componentes de averías. Para poder fijar las pendientes de las cajas de Heisenberg a utilizar siguiendo el criterio de la pendiente definido en 4.2, se necesita conocer las evoluciones de las componentes de avería; en este caso, componentes de excentricidad. Cuando el motor es alimentado desde un inversor, estas evoluciones se pueden determinar si se conoce la evolución de la frecuencia fundamental suponiendo frecuencia de deslizamiento nula, tal y como se explica en 4.1.3. En la presente tesis, con el objetivo de eliminar la necesidad de instalar sensores adicionales, se ha propuesto un método que permite obtener la evolución de la frecuencia fundamental a través de la corriente del estator capturada. Dicho método se ha expuesto en el capítulo 5, apartado 5.1, aplicándolo como ejemplo a una de las corrientes cuya captura se detalla en el apartado anterior (8.1). Así pues, en primer lugar se debe aplicar el método descrito en el capítulo 5 para la obtención de la evolución temporal de la frecuencia de la FC. Esta evolución se muestra trazada en azul en las siguientes figuras: • • • • • Corriente en régimen permanente: Fig. 8.3. Corriente de arranque: Fig. 8.5. Transitorio de disminución de la frecuencia: Fig. 8.7. Transitorio de variación de carga sin compensación del deslizamiento: Fig. 8.9. Transitorio de variación de carga con compensación del deslizamiento: Fig. 8.11. Al despreciar la frecuencia de deslizamiento, la frecuencia de los EH principales viene dada por (4.16) tomando k = 1 : f EH = [1 ± k p ] f FC . Como el motor de inducción tiene dos pares de polos, la frecuencia de los EH principales es respectivamente 1.5 y 0.5 la del FC. Por tanto, se debe multiplicar la evolución de la FC por 1.5 y por 0.5, 328 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. obteniéndose una estimación (pues se desprecia la frecuencia de deslizamiento) de la evolución de los EH. Dichas evoluciones se muestran de color rojo en las correspondientes figuras. Se recuerda que la corriente en régimen permanente pertenece al motor desacoplado de la carga sin excentricidad artificial, estando en los 4 transitorios siguientes acoplado a la carga y con la excentricidad creada. 8.2.3.- Elección de la zona del plano tiempo-frecuencia a analizar. Banda de frecuencia La banda de frecuencia analizada debe cubrir la evolución de la FC y la de los EH principales: E1+ y E1− . Tal y como ya se ha comentado, despreciando la frecuencia de deslizamiento la frecuencia de los EH principales es respectivamente 1.5 y 0.5 la del FC. Teniendo en cuenta que en todos los transitorios capturados la evolución de la frecuencia del FC programada tiene como puntos máximo y mínimo 50 y 30 Hz respectivamente (salvo durante el arranque), las frecuencias máximas y mínimas de los EH resultan ser 75 y 15 Hz aproximadamente. Para que las evoluciones se observen cómodamente, resulta suficiente analizar la banda de frecuencia entre 10 y 90 Hz. Como excepción, durante el transitorio de arranque analizado, la frecuencia del FC empieza obviamente en 0 Hz. Por lo tanto, al analizar la corriente de arranque, el mínimo de la banda de frecuencia analizada es obviamente 0 Hz. Intervalo de tiempo Con respecto al intervalo de tiempo considerado, aunque en cada caso se ha analizado la corriente completa, al presentar los resultados se han recortado los tramos inicial y final donde se encuentra el EB producido por la transformada, el cual no aporta ninguna información adicional. En los casos del régimen permanente, la disminución de la frecuencia y las variaciones de carga (sin y con compensación del deslizamiento), los pequeños tramos inicial y final recortados contienen continuaciones de los correspondientes regímenes permanentes, con lo cual se pueden recortar sin perder información. Sin embargo, en lo que respecta al arranque, no se puede recortar el tramo inicial, pues es una parte importante del arranque. Por lo tanto, en su respectivo resultado se puede observar el EB. 8.2.4.- Selección de la familia de átomos tiempo-frecuencia. Diccionario de partida El siguiente paso es el de seleccionar la familia de átomos tiempo-frecuencia con los cuales se correlacionará la corriente capturada. Recordamos que el conjunto de átomos posibles a utilizar para calcular la transformada (es decir, el diccionario) es el formado por las Frequency B-Splines de parámetro entero m = 2 . Una vez seleccionadas las FBS finalmente a utilizar, estas funciones se recortan temporalmente para reducir el tiempo de cálculo según un parámetro de precisión x = 10.13 % , tal y como se explica en el apartado 6.5. 329 Capítulo 8. Matriz de átomos. Tal y como se explica en el apartado de la guía de aplicación relativo al presente paso (4.7.4), el conjunto de puntos del plano tiempo-frecuencia en los cuales se calculará la correlación forman una matriz donde cada columna se asocia a un instante de tiempo muestreado al capturar la señal a analizar, y cada fila se asocia a una frecuencia analizada, siendo el número de frecuencias analizadas suficientemente elevado y encontrándose uniformemente distribuidas sobre la banda de frecuencia a estudiar. Concretamente, para observar la representación de la densidad de energía, resulta suficiente considerar 200 frecuencias uniformemente distribuidas en la banda de frecuencia a analizar. Sin embargo, para obtener una cuantificación de máxima calidad hace falta considerar aproximadamente 1000 frecuencias cada 50 Hz analizados. En los análisis realizado, se han considerado 1000 frecuencias uniformemente distribuidas en la banda correspondiente. Tal y como se observa en (4.45), para fijar una FBS debemos especificar el punto del plano en el que se encuentra centrada y su ancho de banda. Los puntos del plano han quedado definidos al definir la matriz. La pendiente utilizada en cada uno de dichos puntos se especifica en el siguiente apartado, la cual permitirá calcular el ancho de banda utilizando (4.46). Pendientes de las cajas de Heisenberg utilizadas: consideraciones generales. Al conocer las estimaciones de las evoluciones del FC y de E1+ y E1− , se puede aplicar ya el criterio de la pendiente a cada caso. Pero previamente se realizan algunas observaciones de carácter general. A diferencia de las corrientes analizadas en capítulos anteriores, en los que el motor se alimentaba directamente desde la red, y para las cuales se concluyó que una buena opción era tomar una pendiente distinta a cada frecuencia analizada (permitiendo calcular las correlaciones asociadas a una misma frecuencia con una única convolución), aquí las evoluciones a capturar tienen características muy diferentes y por tanto las conclusiones son diferentes. En primer lugar, los análisis realizados en el presente capítulo tienen como objetivo capturar las evoluciones de tan solo tres componentes: la FC y los EH, las cuales se encuentran relativamente juntas, cubriendo una banda de frecuencia relativamente estrecha. Además, las pendientes de las tres evoluciones en cada instante son bastante parecidas. Así pues, si la FC traza una recta horizontal, los EH también. Y si la FC aumenta o disminuye con una determinada pendiente, los EH también aumentan o disminuyen, con pendientes aproximadamente 1.5 y 0.5 la del FC. Concluyendo, a la hora de analizar estas corrientes parece más adecuado tomar una pendiente distinta en cada instante de tiempo (y la misma para todos las frecuencias en dicho instante). Por ejemplo, si en un determinado instante la pendiente de la FC es nula (traza en ese instante una línea horizontal en el plano tiempo-frecuencia), entonces siguiendo el criterio de la pendiente, resulta adecuado utilizar cajas de Heisenberg con mucha base y poca altura para analizarlo. No resulta adecuado perder por completo la localización temporal anulando por completo la pendiente, siendo un buen valor P = 0.25 Hz s . En ese mismo instante, las pendientes de los EH son también nulas, y 330 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. por tanto resulta adecuado utilizar la misma pendiente para analizarlas. ¿Y en las frecuencias comprendidas entre el FC y los EH? ¿Qué pendientes se utilizan? Como no hay ninguna componente importante a dichas frecuencias, se puede utilizar la misma pendiente, siendo por tanto constante respecto a al frecuencia considerada, una vez fijado el instante de tiempo. Si por el contrario, en un determinado instante la evolución del FC tiene una determinada pendiente, pues su frecuencia está aumentando o disminuyendo, entonces hay que utilizar una caja de Heisenberg cuya pendiente sea idéntica a la de la evolución del FC. Para analizar los EH en ese mismo instante, si se aplica el criterio de la pendiente, las cajas de Heisenberg deben tener pendientes aproximadamente 1.5 y 0.5 la utilizada para analizar la FC. Sin embargo, los resultados son suficientemente buenos si las pendientes utilizadas para analizar dichas componentes son iguales a la utilizada para analizar la FC. Análogamente al ejemplo anterior, en las frecuencias comprendidas entre dichas componentes no hay ninguna otra componente importante a detectar. En definitiva, se puede tomar, para cada instante de tiempo, una única pendiente válida para todas las frecuencias asociadas a dicho instante, e igual a la de la evolución de la FC. Pendientes de las cajas de Heisenberg utilizadas: aplicación a cada corriente. Por ejemplo, en el caso de la corriente en régimen permanente (Fig. 8.3), tanto el FC (evolución azul) como los EH (evoluciones rojas) trazan líneas horizontales en el plano tiempo-frecuencia. Por tanto, la pendiente adecuada en este caso es P = 0.25 Hz s , independientemente de cuál de las tres componentes se analice (pues ambas tienen la misma pendiente) y del instante considerado (pues dichas pendientes no cambian con el tiempo). La evolución de la pendiente de las cajas de Heisenberg consideradas en cada instante se muestra en la Fig. 8.4. Al analizar la corriente de arranque (Fig. 8.5), se observa como la frecuencia de la FC aumenta linealmente de 0 a 50 Hz en aproximadamente 2 segundos (teniendo en cuenta que empieza a aumentar pasados los 0.2 segundos iniciales, y finaliza la evolución a los 2.2 segundos). Por tanto, como la evolución es lineal, la pendiente de la caja adecuada para este tramo es constante e igual a P = 25 Hz s . Para analizar las evoluciones de los EH, las pendientes de las cajas utilizadas deberían multiplicarse por 1.5 y 0.5 respectivamente. Sin embargo, tal y como se ha comentado, nos contentamos con aplicar la misma pendiente utilizada para el FC. Así pues, en los distintos instantes que dura el arranque, la pendiente en cada uno de ellos es igual a P = 25 Hz s , independientemente de la frecuencia considerada. Y una vez ha terminado el arranque, como las evoluciones se tornan horizontales, la pendiente de las cajas utilizadas debe caer a un valor 10 veces menor P = 0.25 Hz s . Este cambio en la pendiente no se produce de forma instantánea: una vez finalizado el arranque en 2.2 segundos, la pendiente de las cajas disminuye linealmente con el tiempo alcanzando el valor propio de un régimen permanente a los 3 segundos de iniciarse el transitorio. Sin embargo, para que la transición no sea demasiado brusca, en vez de tomar una pendiente P = 0.25 Hz s en régimen permanente se toma P = 1 Hz s . En definitiva, la evolución de la pendiente de las cajas de Heisenberg consideradas en cada instante de tiempo se muestra en la Fig. 8.6. 331 Capítulo 8. Fig. 8. 3 Análisis de la corriente en régimen permanente: estimación de la evolución temporal de las frecuencias de la FC (azul) y los EH (rojo). Fig. 8. 4 Análisis de la corriente en régimen permanente: pendiente de las cajas de Heisenberg a utilizar en función del instante de tiempo analizado. 332 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 5 Análisis de la corriente de arranque: estimación de la evolución temporal de las frecuencias de la FC (azul) y los EH (rojo). Fig. 8. 6 Análisis de la corriente de arranque: pendiente de las cajas de Heisenberg a utilizar en función del instante de tiempo analizado. 333 Capítulo 8. Al analizar el transitorio en el que la frecuencia asignada disminuye (Fig. 8.7), se observa que la frecuencia del FC es constante e igual a 50 Hz durante los tres primeros segundos del transitorio, disminuyendo linealmente con el tiempo hasta 33 Hz en 1 segundo, quedándose en dicho valor hasta el final. Para analizar el segundo durante el cual la frecuencia del FC disminuye linealmente, se toma una caja de pendiente igual a la de su evolución: P = ( 50 − 33) 1 = 17 Hz s . Para analizar los regímenes permanentes anterior y posterior se toma una caja de pendiente P = 1 Hz s (no utilizando P = 0.25 Hz s con el objetivo de que la transición entre ambas pendientes no sea demasiado brusca). Así pues, se toma una pendiente P = 1 Hz s durante los dos primeros segundos (Fig. 8.8), evolucionando linealmente con el tiempo hasta llegar a P = 17 Hz s cuando se cumplen los tres segundos de duración, manteniéndose en dicho valor durante el segundo que dura la disminución de frecuencia y volviendo linealmente en un segundo al valor correspondiente al régimen permanente. Como en los ejemplos anteriores, las pendientes utilizadas para analizar la evolución del FC en cada instante son las mismas que las utilizadas en las otras frecuencias. Análogamente se obtienen las pendientes a utilizar al analizar las variaciones de carga sin (Fig. 8.9) y con (Fig. 8.11) compensación del deslizamiento. En el primer caso las pendientes son las mismas que si se trata de un régimen permanente (Fig. 8.10), pues la evolución del FC es una recta horizontal y aunque las evoluciones de los EH no son completamente rectilíneas debido a los cambios en el deslizamiento provocados por la variación de carga, esta particularidad no se ha tenido en cuenta al trazarlas en la Fig. 8.9, pues en esta etapa de estimación de la evolución de las componentes se ha supuesto precisamente frecuencia de deslizamiento nula. Así pues, teniendo en cuenta la Fig. 8.9, las pendientes de las cajas de Heisenberg utilizadas en cada instante de tiempo se muestran en la Fig. 8.10. En el segundo caso las pendientes de las cajas durante la variación de carga se ajustan a las de la evolución del FC que aparece para compensar el cambio en el deslizamiento (Fig. 8.11). Concretamente, la frecuencia del FC evoluciona de 50.5 a 53 en 3 segundos según una tendencia aproximadamente lineal. Por tanto, la pendiente de las cajas durante este transitorio es constante e igual a P = ( 53 − 50.5) 3  0.83 Hz s. En los regímenes posterior y anterior se utilizan pendientes propias de un régimen permanente: P = 0.25 Hz s . A los 3 segundos la pendiente cambia linealmente con respecto al tiempo partiendo de P = 0.25 Hz s y llegando a P  0.83 Hz s a los cuatro segundos. A los 7 segundos, cuando la variación de carga ya ha finalizado, la pendiente vuelve a bajar linealmente, alcanzando otra vez el valor propio del régimen permanente a los 8 segundos, manteniéndose así hasta el final (Fig. 8.12). 8.2.5.- Cálculo y representación gráfica de la función de densidad tiempo-frecuencia. Una vez fijada la familia de átomos tiempo-frecuencia, el siguiente paso es calcular la correlación habitual entre los elementos de la familia y la señal a analizar: +∞ h,ψ tc , f c =  h ( t )ψ t∗c , f c ( t ) dt −∞ 334 (8.1) Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 7 Análisis del transitorio de disminución de la frecuencia: estimación de la evolución temporal de las frecuencias de la FC (azul) y los EH (rojo). Fig. 8. 8 Análisis del transitorio de disminución de la frecuencia: pendiente de las cajas de Heisenberg a utilizar en función del instante de tiempo analizado. 335 Capítulo 8. Fig. 8. 9 Análisis del transitorio de variación de carga sin compensación del deslizamiento: estimación de la evolución temporal de las frecuencias de la FC (azul) y los EH (rojo). Fig. 8. 10 Análisis del transitorio de variación de carga sin compensación del deslizamiento: pendiente de las cajas de Heisenberg a utilizar en función del instante de tiempo analizado. 336 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 11 Análisis del transitorio de variación de carga con compensación del deslizamiento: estimación de la evolución temporal de las frecuencias de la FC (azul) y los EH (rojo). Fig. 8. 12 Análisis del transitorio de variación de carga con compensación del deslizamiento: pendiente de las cajas de Heisenberg a utilizar en función del instante de tiempo analizado. 337 Capítulo 8. Finalmente, se calculan las densidades de energía en cada punto del plano analizado: ρ h ( tc , f c ) = h,ψ t , f c 2 c (8.2) Como los átomos de la familia están centrados en una matriz de puntos en el plano tiempo-frecuencia, al aplicar (8.2) obtenemos como resultado una matriz de densidades de energía. Las densidades de energía obtenidas se representan en 2D en el plano tiempo-frecuencia localizando los puntos de la matriz de resultados que contienen los valores máximo y mínimo, asignándoles respectivamente los dos extremos de una determinada escala de colores, para proceder seguidamente a asignar colores intermedios al resto de resultados en función de sus valores relativos a dichos extremos. Concretamente, la escala de colores utilizada es la escala ‘jet’ de MatLab y se han considerando 1024 colores. Por otra parte, para poder observar mejor las evoluciones de las componentes de menor amplitud, mejorando la visualización de los resultados obtenidos, se ha aplicado escala logarítmica. En definitiva, al aplicar la metodología con las pendientes indicadas y según lo explicitado en los subapartados anteriores, los resultados son los mostrados en el siguiente conjunto de figuras: • • • • • Corriente en régimen permanente: Fig. 8.13. Corriente de arranque: Fig. 8.17. Transitorio de disminución de la frecuencia: Fig. 8.21. Transitorio de variación de carga sin compensación del deslizamiento: Fig. 8.25. Transitorio de variación de carga con compensación del deslizamiento: Fig. 8.29. Las tres figuras siguientes que acompañan a cada una de las recién mencionadas muestran los resultados de aplicar, a cada corriente, el método de cuantificación explicado en 8.2.7. 8.2.6.- Aplicación de técnicas de mejora de la visualización. Las figuras obtenidas en el apartado anterior se han generado utilizando escala logarítmica. Por lo demás, evitando alargar demasiado el capítulo, en el presente apartado no se utilizan otras de las técnicas propuestas en la tesis para mejorar la visualización, pues aplicando escala logarítmica ya se observan claramente las evoluciones de las componentes. 8.2.7.- Cuantificación. Para cuantificar la presencia de excentricidad en el motor cuya corriente del estator se analiza, se ha utilizado el método expuesto en el apartado 4.6.2. Se recuerda que, en el primer paso del método se obtiene la evolución real de los EH. Para ello se considera una banda de frecuencia de 2 Hz de amplitud alrededor de la estimación inicial de su evolución. Seguidamente se calcula, para cada instante, el máximo dentro de las respectivas bandas. Las figuras 8.14, 8.18, 8.22, 8.26 y 8.30 (vinculada cada figura a uno de los transitorios analizados), muestran las evoluciones temporales del FC 338 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. (negro), las bandas de frecuencia que deben contener las evoluciones de E1+ y E1− (azul y verde respectivamente), y las evoluciones de dichas componentes finalmente calculadas (trazadas en rojo). El siguiente paso consiste en localizar en la matriz resultado de calcular la AST las evoluciones definitivas de E1+ y E1− trazadas en rojo. El resultado se muestra en tres dimensiones en las figuras 8.15, 8.19, 8.23, 8.27 y 8.31. Finalmente, prescindiendo de la información frecuencial, se obtiene para cada instante (figuras 8.16, 8.20, 8.24, 8.28 y 8.32), la densidad de energía a lo largo de las evoluciones del FC (negro), el E1+ (azul) y el E1− (verde). Las medias de las evoluciones de dichas densidades son los índices que se utilizan para cuantificar la presencia de la avería (Tabla 8.1). Tan solo en el caso del arranque se han suprimido los primeros 1.4 segundos antes de calcular la media, pues durante esta parte las evoluciones de las componentes de avería se confunden con la del FC, falseando por tanto el resultado. El límite a partir del cual se ha calculado la media se ha indicado con una línea vertical roja en la Fig. 8.20. Tabla. 8. 1 Índices que cuantifican la presencia de los dos componentes vinculados a la avería, para cada uno de los transitorios analizados. E1+ (dB) 58.04 34.43 38.26 28.39 31.30 Transitorio Régimen permanente Arranque Disminución de la frecuencia Variación de carga sin compensación Variación de carga con compensación 339 E1− (dB) 59.08 32.29 37.03 25.75 28.84 Capítulo 8. Fig. 8. 13 AST de la corriente en régimen permanente. La pendiente de las cajas utilizadas varía con el instante analizado según lo indicado en la Fig. 8.4. Fig. 8. 14 Análisis de la corriente en régimen permanente: Estimación de la evolución de la FC (azul), bandas de frecuencia alrededor de la frecuencia estimada de los EH (azul EH superior y verde EH inferior) y evoluciones de los EH finalmente calculadas (rojo). 340 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 15 Visualización tridimensional del acople de los resultados de las figuras 8.13 y 8.14: localización de los puntos de la evolución temporal de los EH en la matriz que describe la densidad de energía. Fig. 8. 16 Análisis de la corriente en régimen permanente: Evolución temporal de la densidad de energía vinculada al FC (negro, coincidente con 0 dB) al EH superior (azul) y al EH inferior (verde). 341 Capítulo 8. Fig. 8. 17 AST de la corriente de arranque. La pendiente de las cajas utilizadas varía con el instante analizado según lo indicado en la Fig. 8.6. Fig. 8. 18 Análisis de la corriente de arranque: Estimación de la evolución de la FC (azul), bandas de frecuencia alrededor de la frecuencia estimada de los EH (azul EH superior y verde EH inferior) y evoluciones de los EH finalmente calculadas (rojo). 342 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 19 Visualización tridimensional del acople de los resultados de las figuras 8.17 y 8.18: localización de los puntos de la evolución temporal de los EH en la matriz que describe la densidad de energía. Fig. 8. 20 Análisis de la corriente de arranque: Evolución temporal de la densidad de energía vinculada al FC (negro) al EH superior (azul) y al EH inferior (verde). 343 Capítulo 8. Fig. 8. 21 AST del transitorio de disminución de la frecuencia. La pendiente de las cajas utilizadas varía con el instante analizado según lo indicado en la Fig. 8.8. Fig. 8. 22 Análisis del transitorio de disminución de la frecuencia: Estimación de la evolución de la FC (azul), bandas de frecuencia alrededor de la frecuencia estimada de los EH (azul EH superior y verde EH inferior) y evoluciones de los EH finalmente calculadas (rojo). 344 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 23 Visualización tridimensional del acople de los resultados de las figuras 8.21 y 8.22: localización de los puntos de la evolución temporal de los EH en la matriz que describe la densidad de energía. Fig. 8. 24 Análisis del transitorio de disminución de la frecuencia: Evolución temporal de la densidad de energía vinculada al FC (negro) al EH superior (azul) y al EH inferior (verde). 345 Capítulo 8. Fig. 8. 25 AST del transitorio de variación de carga sin compensación del deslizamiento. La pendiente de las cajas utilizadas varía con el instante analizado según lo indicado en la Fig. 8.10. Fig. 8. 26 Análisis del transitorio de variación de carga sin compensación del deslizamiento: Estimación de la evolución de la FC (azul), bandas de frecuencia alrededor de la frecuencia estimada de los EH (azul EH superior y verde EH inferior) y evoluciones de los EH finalmente calculadas (rojo). 346 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 27 Visualización tridimensional del acople de los resultados de las figuras 8.25 y 8.26: localización de los puntos de la evolución temporal de los EH en la matriz que describe la densidad de energía. Fig. 8. 28 Análisis del transitorio de variación de carga sin compensación del deslizamiento: Evolución temporal de la densidad de energía vinculada al FC (negro) al EH superior (azul) y al EH inferior (verde). 347 Capítulo 8. Fig. 8. 29 AST del transitorio de variación de carga con compensación del deslizamiento. La pendiente de las cajas utilizadas varía con el instante analizado según lo indicado en la Fig. 8.12. Fig. 8. 30 Análisis del transitorio de variación de carga con compensación del deslizamiento: Estimación de la evolución de la FC (azul), bandas de frecuencia alrededor de la frecuencia estimada de los EH (azul EH superior y verde EH inferior) y evoluciones de los EH finalmente calculadas (rojo). 348 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. Fig. 8. 31 Visualización tridimensional del acople de los resultados de las figuras 8.29 y 8.30: localización de los puntos de la evolución temporal de los EH en la matriz que describe la densidad de energía. Fig. 8. 32 Análisis del transitorio de variación de carga con compensación del deslizamiento: Evolución temporal de la densidad de energía vinculada al FC (negro) al EH superior (azul) y al EH inferior (verde). 349 Capítulo 8. 8.3.- Análisis de los resultados. Diagnóstico. En el presente apartado se analizan los resultados obtenidos formulando un diagnóstico. En los dos párrafos siguientes se comentan a grandes rasgos los resultados. Seguidamente se analizan con más detalle cada una de las evoluciones captadas y sus resultados asociados. Finalmente se extraen las conclusiones. Así pues, en primer lugar, al estudiar el resultado relativo al análisis de la corriente en régimen permanente (Fig. 8.13), no se observa ninguna de las dos componentes principales provocadas por la excentricidad. Esta ausencia también se refleja de forma cuantitativa al considerar la media de la densidad de energía asociada a dichas componentes, pues el resultado obtenido es el propio de un cierto nivel de ruido: 58.04 dB en el caso de E1+ y 59.08 dB en el caso de E1− . Se concluye que, según la metodología, el motor está sano. El resultado del diagnóstico es correcto, pues esta corriente se tomó con el motor desacoplado de la carga, sin ningún tipo de excentricidad creada. Por tanto se puede concluir además que la excentricidad inherente del motor analizado es prácticamente nula. Por otra parte, en los resultados relativos a las cuatro corrientes transitorias restantes analizadas (figuras 8.17, 8.21, 8.25 y 8.29), se observa, aparte de la evolución de la FC, las evoluciones de las dos componentes principales causadas por la excentricidad, lo cual indica que dicha avería se encuentra presente en el motor. Al mismo tiempo, se ha cuantificado su presencia al calcular el valor medio de la densidad de energía a lo largo de las trayectorias de E1+ y de E1− , mostrándose los resultados en la tabla 8.1. Todos los resultados indican un elevado nivel energético asociado a E1+ y E1− , con una diferencia mínima con el caso sano de 20 dB. Con respecto a estas cuatro corrientes transitorias, el resultado del diagnóstico es también correcto, pues han sido capturadas creando una excentricidad en el motor. 8.3.1.- Corriente en régimen permanente. El resultado de aplicar la metodología a la corriente en régimen permanente se muestra en las figuras 8.13 a 8.16. En la Fig. 8.13 tan solo se observa la trayectoria de la FC, cuya frecuencia es constante, lo cual es normal pues el motor se encuentra funcionando en régimen permanente. Se han utilizado cajas de Heisenberg de muy poca pendiente, obteniendo una gran resolución en frecuencia, lo cual permite observar con precisión la frecuencia de la FC: 53 Hz. Por otra parte, en la banda de frecuencia analizada, las únicas componentes que pueden aparecer aparte de la FC son las provocadas por fallos como los EH vinculados a la excentricidad. Como no aparecen en el resultado, se puede concluir que el motor está sano. No obstante, la metodología se aplica completa, trazándose en la Fig. 8.14 la estimación de la evolución de la FC (negro), así como las bandas de frecuencia que contienen la evolución de los EH en caso de estar presentes en la corriente (azul y verde). Evidentemente, como los EH no se encuentran presentes en la señal, la hipotética estimación final de su evolución trazada en rojo, está en realidad vinculada a los puntos con mayor nivel de ruido dentro de las respectivas bandas. 350 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. En la Fig. 8.15 se observa la evolución de la FC en tres dimensiones, con una densidad de energía mucho mayor que el resto de puntos del plano, estando las evoluciones de los EH vinculadas a un determinado nivel de ruido. Finalmente, la evolución de la densidad de energía de la FC, trazada en negro en la Fig. 8.16, se confunde con el eje de abscisas, pues al aplicar escala logarítmica y tratarse del máximo valor en todo instante, su densidad de energía asociada es de 0 dB en todo instante. Por otra parte, las densidades de energía asociadas a los hipotéticos EH (verde y azul), se encuentran alrededor de 60 dB, oscilando en función del nivel de ruido en cada instante considerado. 8.3.2.- Corriente de arranque. En la Fig. 8.17, resultado de analizar la corriente de arranque, se observan las trayectorias de la FC y los EH. Las evoluciones captadas son exactamente las que se esperaban plasmadas en la Fig. 8.5. Además, al iniciarse el régimen permanente se pueden ver un conjunto de componentes próximas a la FC evolucionando paralelamente a ella. Dichas componentes son introducidas por el inversor debido a que la compensación del deslizamiento se encuentra activada. Por otra parte, se observa claramente el efecto que produce sobre los resultados el cambio en las dimensiones de las cajas de Heisenberg utilizadas. Durante el aumento de la frecuencia asignada, las cajas tienen una pendiente igual a la de la FC, capturando su energía de la mejor forma posible. Sin embargo, dicha pendiente es demasiado elevada para analizar el régimen permanente. Así pues en 2.2 segundos, cuando ha terminado el arranque, la pendiente de la caja decrece, y por tanto la banda de frecuencia en la que se extiende la presencia de la FC disminuye hasta alcanzar una buena localización en frecuencia. En definitiva, la componente refleja su presencia sobre el plano entre 2.2 y 3 segundos en forma de cuña, debido a que a medida que avanza el tiempo, la pendiente de la caja utilizada disminuye, aumentando la resolución en frecuencia, disminuyendo por tanto el ancho de banda en frecuencia que ocupa la componente. Esto permite determinar con precisión la frecuencia de la FC, resultando ser ligeramente superior a 50 Hz como consecuencia de que la compensación del deslizamiento se encuentra activada. Idénticos comentarios se pueden realizar respecto a la captación de las evoluciones de los EH. Si se hubiera utilizado por ejemplo la STFT, podríamos captar bien el arranque y tener muy mala resolución en frecuencia durante el régimen permanente, o captar muy bien la frecuencia durante el permanente y emborronar por completo la información durante el arranque. Con respecto a la estimación final de las evoluciones de los EH mostradas en la Fig. 8.18, se observa cómo, hasta pasados los primeros 1.5 segundos no se captan adecuadamente, pues los EH se encuentran demasiado cerca de la FC, solapándose y confundiéndose con ella: es por ello que los máximos locales calculados durante estos primeros instantes, los cuales deberían estar asociados a las evoluciones de los EH, se encuentran en los extremos de las respectivas bandas, lo más próximos posibles a la evolución de la FC, vinculados a esta última. En la Fig. 8.17 es a partir del primer segundo que se empiezan a observar los EH, pero hasta llegar a 1.4 segundos el algoritmo de cálculo de máximos locales aplicado para obtener la estimación final de las evoluciones obtiene resultados asociados al FC. Esto mismo se observa muy bien en la Fig. 8.19 (donde se han acoplado en 3D los resultados de las figuras 8.17 y 8.18): las 351 Capítulo 8. evoluciones de los EH al principio se solapan con la evolución de la FC, siendo las densidades de energía asociadas significativas de la FC y no de los EH. En el resto de puntos, donde los máximos calculados sí están asociados a la presencia de los EH, los resultados se encuentran más cerca del FC debido a que en funcionamiento motor la frecuencia de deslizamiento es ligeramente superior a cero, desplazando la estimación final con respecto a la inicial del centro de la banda hacia la FC. En la Fig. 8.19, aparte de los 1.4 primeros segundos, la metodología obtiene adecuadamente las evoluciones de las densidades de energía asociadas a la FC y los EH, representadas a su vez en 2D en la Fig. 8.20. En esta última figura se observa la evolución de la densidad de energía de la FC (negro). Durante los primeros 0.2 segundos, la corriente es todavía muy baja y la densidad de energía tiene valores propios de un nivel de ruido. Durante el arranque, (entre 0.2 y 2.2 segundos), la densidad de energía de la FC es relativamente constante, con una ligera pendiente ascendente. Cuando termina el arranque a los 2.2 segundos y se empiezan a cambiar las dimensiones de las cajas utilizadas, la densidad de energía aumenta pues la caja adquiere una forma más adecuada para capturar la energía de la componente. Una vez se ha terminado ese proceso transitorio de cambio de las dimensiones de las cajas, concretamente a los 3 segundos, la densidad de energía se estabiliza en un determinado valor, el máximo obtenido al calcular la transformada, y que por tanto tiene asignado en escala logarítmica el valor de 0 dB. Por otra parte, la densidad de energía de los EH durante los primeros 1.4 segundos, marcados por una línea vertical roja, está más asociada a la presencia del FC que a la suya (lo cual se observa muy bien en la Fig. 8.19). Una vez terminados estos primeros instantes, existen unas ciertas oscilaciones durante el arranque, y una vez pasados los primeros 2.2 segundos, hay una tendencia ascendente debido al cambio en la pendiente de las cajas, estabilizándose al llegar a 3 segundos, siendo la densidad de energía de E1− un tanto superior. Los valores medios de las funciones azul ( E1+ ) y verde ( E1− ) representadas en la Fig. 8.20 descartando los primeros 1.4 segundos son (tal y como se indica en la Tabla 8.1): 34.43 dB y 32.29 dB respectivamente. 8.3.3.- Transitorio de variación de la frecuencia. El ensayo en el que se ha disminuido la frecuencia asignada desde 50 hasta 32.9 Hz se ha realizado sin compensar la variación del deslizamiento. Es por ello que en el resultado correspondiente (Fig. 8.21), se observan las evoluciones de la FC y los EH, pero no hay ninguna otra componente introducida por el inversor. Estas evoluciones obtenidas son las previstas en la Fig. 8.7. Tal y como se ha explicado, se han tomado cajas propias de un análisis en régimen permanente hasta los 2 primeros segundos y después de los 5 (Fig. 8.8). Entre 2 y tres hay una evolución creciente de la pendiente, adaptándose para obtener adecuadamente la evolución durante la disminución de la frecuencia. Es por ello que se observa cómo se pierde paulatinamente la resolución en frecuencia y tanto la FC como los EH ocupan una banda de frecuencia de amplitud mayor alrededor de su frecuencia real. Seguidamente se capta la disminución de la frecuencia, capturando la energía de las componentes de forma óptima, y a partir de los 3 segundos se recupera la resolución en frecuencia adecuada para volver a analizar las evoluciones horizontales. Si se hubiera utilizado la STFT, se podrían haber captado las frecuencias en permanente con elevada 352 Diagnóstico de excentricidad cuando el motor es alimentado por un inversor. precisión pero emborronando la trayectoria con frecuencia decreciente o se podría haber obtenido adecuadamente dicha disminución de frecuencia renunciando a la resolución durante los permanentes anterior y posterior. En la Fig. 8.22 se observan trazadas de color rojo la estimación final de las evoluciones de los EH, las cuales se encuentran prácticamente en el centro de la banda de frecuencia considerada inicialmente pues el motor está poco cargado y en estas condiciones el suponer frecuencia de deslizamiento nula resulta muy aproximado. Al acoplar este resultado con el mostrado en la Fig. 8.21 trazando ambos en 3D se obtiene la Fig. 8.23. Aquí se observa que la estimación final de las evoluciones de los EH debe ser muy precisa para obtener correctamente su densidad de energía asociada pues a poco que aparezca una desviación, en vez de medir su densidad de energía se mide el nivel de ruido.	17,939
https://openalex.org/W2913422347	OpenAlex	Open Science	CC-By	2,018	Cyto-morphological investigation of cellular content of peripancreatic liquid accumulations in estimation of the clinical course severity in an acute complicated pancreatitis	В. П. Андрющенко	English	Spoken	2,767	11,064	Cyto–morphological investigation of cellular content of peripancreatic liquid accumulations in estimation of the clinical course severity in an acute complicated pancreatitis To study up a character and peculiarities of cyto–morphological changes in cellular material of the peripancreatic liquid accumulations (LA) as a criterion of objective estimation of the clinical course severity in an acute complicated pancre- atitis (ACP). Objective. To study up a character and peculiarities of cyto–morphological changes in cellular material of the peripancreatic liquid accumulations (LA) as a criterion of objective estimation of the clinical course severity in an acute complicated pancre- atitis (ACP). Objective. To study up a character and peculiarities of cyto–morphological changes in cellular material of the peripancreatic liquid accumulations (LA) as a criterion of objective estimation of the clinical course severity in an acute complicated pancre- atitis (ACP) ( ) Маterials and methods. Results of cyto–morphological investigation of the peripancreatic LA sediment, obtained while do- ing interventional ultrasonography in 51 (88%) observations and videolaparoscopy – in 7 (12%) observations in 58 patients with an ACP, were analyzed. ( ) Маterials and methods. Results of cyto–morphological investigation of the peripancreatic LA sediment, obtained while do- ing interventional ultrasonography in 51 (88%) observations and videolaparoscopy – in 7 (12%) observations in 58 patients with an ACP, were analyzed. Маterials and methods. Results of cyto–morphological investigation of the peripancreatic LA sediment, obtained while do- ing interventional ultrasonography in 51 (88%) observations and videolaparoscopy – in 7 (12%) observations in 58 patients with an ACP, were analyzed. Results. Of intraabdominal LA (n=37) observations in 10 (27 %) of them a domination of proliferating cells of mesothelium with their polymorphism was revealed, while in 15 (41%) – degenerative–dystrophic changes like a bulging of cytoplasm, appearance of large «signet–ring like» cells, in 12 (32%) – аtipically changed cells of mesothelium with signs of anizocytosis and anizonucleosis, as well as cyto–morphological changes like in neoplastic process. In retroperitoneal localization of LA (n=21) a more essential changes in the granulocytes nuclei were registered, including, in 9 (43%) observations their structure have got irreversible disorders: karyopycnosis, karyorexis and karyolysis. The depth of structural cellular changes in mesoteliocytes and neutrophil granulocytes have depended upon the ACP severity. p g y p p y Conclusion. Determination of depth of the cellular content cytomorphological disorders in the LA sediment may serve an objective criterion in the ACP severity estimation. j y eywords: acute complicated pancreatitis; liquid accumulations; cytological investigations. ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia Klinichna khirurhiia. 2018 November;85(11):9-12. DOI: 10.26779/2522-1396.2018.11.09 Cyto–morphological investigation of cellular content of peripancreatic liquid accumulations in estimation of the clinical course severity in an acute complicated pancreatitis Реферат Мета. Вивчити характер та особливості цитоморфологічних змін клітинного матеріалу осаду перипанкреатичних рі- динних скупчень (РС) як критерію об’єктивної оцінки тяжкості клінічного перебігу гострого ускладненого панкреа- титу (ГУП). і і ф у ( ) Матеріали і методи. Проаналізовано результати цитоморфологічного дослідження осаду перипанкреатичних РС, добутого під час виконання інтервенційної ультрасонографії – 51 (88%) спостереження та відеолапароскопії – 7 (12%) спостережень у 58 хворих із ГУП. Матеріали і методи. Проаналізовано результати цитоморфологічного дослідження осаду перипанкреатичних РС, добутого під час виконання інтервенційної ультрасонографії – 51 (88%) спостереження та відеолапароскопії – 7 (12%) спостережень у 58 хворих із ГУП. Результати. У разі внутрішньочеревних РС (n=37) у 10 (27 %) спостереженнях виявлено домінування проліферуючих клітин мезотелію з їх поліморфізм, у 15 (41%) – дегенеративно–дистрофічні зміни у вигляді вип ячувань цитоплазми, появу великих «перснеподібних» клітин, у 12 (32%) – атипово змінені клітини мезотелію з проявами анізоцитозу та ані- зонуклеозу, а також цитоморфологічні зміни, подібні до неопластичного процесу. У разі позаочеревинної локалізації РС (n=21) фіксували істотніші зміни ядер гранулоцитів, зокрема, у 9 (43%) спостереженнях їх структура мала незворотні порушення у вигляді явищ каріопікнозу, каріорексису та каріолізису. Глибина структурних змін клітин мезотеліоцитів та нейтрофільних гранулоцитів залежала від ступеня тяжкості ГУП. Результати. У разі внутрішньочеревних РС (n=37) у 10 (27 %) спостереженнях виявлено домінування проліферуючих клітин мезотелію з їх поліморфізм, у 15 (41%) – дегенеративно–дистрофічні зміни у вигляді вип ячувань цитоплазми, появу великих «перснеподібних» клітин, у 12 (32%) – атипово змінені клітини мезотелію з проявами анізоцитозу та ані- зонуклеозу, а також цитоморфологічні зміни, подібні до неопластичного процесу. У разі позаочеревинної локалізації РС (n=21) фіксували істотніші зміни ядер гранулоцитів, зокрема, у 9 (43%) спостереженнях їх структура мала незворотні порушення у вигляді явищ каріопікнозу, каріорексису та каріолізису. Глибина структурних змін клітин мезотеліоцитів та нейтрофільних гранулоцитів залежала від ступеня тяжкості ГУП. р ф р у у Висновки. Визначення глибини цитоморфологічних порушень клітинного складу осаду РС може слугувати об’єктив- ним критерієм оцінки ступеня тяжкості ГУП. р ф р у у Висновки. Визначення глибини цитоморфологічних порушень клітинного складу осаду РС може слугувати об’єктив- ним критерієм оцінки ступеня тяжкості ГУП. Висновки. Визначення глибини цитоморфологічних порушень клітинного складу осаду РС може слугувати об’єктив- ним критерієм оцінки ступеня тяжкості ГУП. р р у ючові слова: гострий ускладнений панкреатит; рідинні скупчення; цитологічне дослідження. Abstract Objective. Klinichna khirurhiia. 2018 November; 85(11) Materials and methods studies The analytical stage provided for a cytological study of na- tive and stained samples. Samples were Pappenheim–stained. Thus, 10 (27%) studies showed dominance in cytological specimens of proliferating mesothelium, characterized by large mesotheliocytes, nuclei and their polymorphism. 15 (41%) cases showed degenerative–dystrophic changes in both cy- toplasm and mesotheliocytes in the form of cytoplasmic pro- trusions, its vacuolation, appearance of foaminess, sometimes with large castration cells. 12 (32%) studies showed the pres- ence of atypically altered mesothelium cells with anisocyto- sis and anisonucleosis, as well as cytomorphological changes similar to neoplastic process (fig. 1, 2). Cytomorphological abnormalities of peripancreatic FC con- tent with their localization in retroperitoneal space (21 ob- servations), in the absence of direct contact of the pancreatic fluid with peritoneum, were most significant in neutrophilic granulocytes. Thus, all studies showed changes in these cells cytoplasm in the form of toxic grains and vacuolations both in individual cells and most neutrophil granulocytes, inten- sive basophilia, decreased specific graininess and appearance of circular colourless spots, which reflected the initial process of degenerative–dystrophic cell changes. p p pp Fluid collections content was transferred into test tube or other, more spacious, clean, dry labware with 5% sodium cit- rate solution at a rate of 2 – 5 ml per 100 ml of the test mate- rial in order to stabilize it and prevent the destruction of cel- lular elements. With a considerable volume of fluid material, it was centrifuged at 1500 rpm for 5 – 10 minutes, after which supernatant was discharged, and 4–6 preparations were re- ceived from the precipitate with drying for 1 hour and subse- quent staining. Further study was performed using BIOLAM and MIKMED–5 light microscopes with a magnification of î40, î90, î100. Microscopy was applied to both native and stained specimen with small and large microscope magnifi- cations (immersion system). The obtained study results were processed using variation statistics of t–test, Fischer test and Ȥð. However, more significant changes were observed in gran- ulocyte nuclei. Thus, in 9 (43%) cases, irreversible structural damages were observed characteristic of the gradually–se- quential stages of death, namely karyopyknosis, karyoclasis and karyolysis (fig. 3, 4). An analysis was performed of cytomorphological changes in mesotheliocytes and neutrophil granulocytes with intra– abdominal and retroperitoneal FC location depending on the severity of the clinical course of disease according to AP clas- sification criteria (Atlanta, 2012) [1] (table 1, 2). Materials and methods studies Cytomorphological study of peripancreatic FC sediment produced during interventional ultrasonography in 51 (88%) and video–laparoscopy in 7 (12%) observations was per- formed in 58 patients who were under the inpatient treatment in the City Pancreatic Centre at the Academic Department of General Surgery of Danylo Halytsky Lviv National Medical University, with a confirmed ACP diagnosis (based on clinical, laboratory–biochemical, radiological and instrumental meth- ods). The age of hospitalised patients ranged from 22 to 74 years. 36 female (33%), 73 male (67%). The cytological study of pancreatic and peripancreatal FC contents provided for the implementation of two stages – pre–analytical and analyti- cal. The first one included qualified collection and transpor- tation of study material, which was important for further re- sult. The analytical stage provided for a cytological study of na- tive and stained samples. Samples were Pappenheim–stained. Fluid collections content was transferred into test tube or other, more spacious, clean, dry labware with 5% sodium cit- rate solution at a rate of 2 – 5 ml per 100 ml of the test mate- rial in order to stabilize it and prevent the destruction of cel- lular elements. With a considerable volume of fluid material, it was centrifuged at 1500 rpm for 5 – 10 minutes, after which supernatant was discharged, and 4–6 preparations were re- ceived from the precipitate with drying for 1 hour and subse- quent staining. Further study was performed using BIOLAM and MIKMED–5 light microscopes with a magnification of î40, î90, î100. Microscopy was applied to both native and stained specimen with small and large microscope magnifi- cations (immersion system). The obtained study results were processed using variation statistics of t–test, Fischer test and Ȥð. Cytomorphological study of peripancreatic FC sediment produced during interventional ultrasonography in 51 (88%) and video–laparoscopy in 7 (12%) observations was per- formed in 58 patients who were under the inpatient treatment in the City Pancreatic Centre at the Academic Department of General Surgery of Danylo Halytsky Lviv National Medical University, with a confirmed ACP diagnosis (based on clinical, laboratory–biochemical, radiological and instrumental meth- ods). The age of hospitalised patients ranged from 22 to 74 years. 36 female (33%), 73 male (67%). The cytological study of pancreatic and peripancreatal FC contents provided for the implementation of two stages – pre–analytical and analyti- cal. The first one included qualified collection and transpor- tation of study material, which was important for further re- sult. ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia and proinflammatory cytokines on cellular elements resulted in pathological changes of all cellular elements as determined in cytological specimens. However, the most significant and indicative were the changes in FC mesotheliocytes with in- tra–abdominal location (37 observations), since these cells, being the main structural elements of serous membranes, in particular parietal and visceral peritoneum, were in direct con- tact with enzymatically active pancreatic exudate. characteristics [2,3]. Is it possible to assess FC content in de- termining the severity of the clinical course of disease? characteristics [2,3]. Is it possible to assess FC content in de- termining the severity of the clinical course of disease? Purpose studies: to study the character and peculiarities of cytomorphological changes of cellular sediment material of flu- id collections (FC) as criterion for objective assessment of the clinical course severity in acute complicated pancreatitis (ACP). Materials and methods studies As can be seen from the data presented in tables, in moderately severe ACP, in contrast to the severe clinical course of the disease, the cytological presentation of intra–abdominal FC was characterized by damage to mesotheliocytes structure in the form of their degenerative–dystrophic changes – in 14 Cyto–morphological investigation of cellular content of peripancreatic liquid accumulations in estimation of the clinical course severity in an acute complicated pancreatitis The updated classification of acute pancreatitis (AP) (Atlanta, 2012) provides an in–depth description of local disease com- plications and, first of all, pancreatic fluid collections (FC) [1]. Thus, the following FC types distinguished. 1. Acute peripancre- atic FC with aseptic contents without signs of pancreas necro- sis and parapancreatic fat, which are aseptic (Acute peripancre- atic flud collection – APFC). 2. Acute necrotic FC with different amounts of both fluid and necrotic tissue fragments associated with the necrosis in pancreas parenchyma and/or parapancre- atic tissue (Acute necrotic collection – ANC). 3. Post–necrotic infected pancreatic and/or parapancreatic FC with contents in the form of a fluid component/purulence and necrotic tis- sue/detritus predominantly infected (Post–necrotic pancreat- ic/retropancreatic flud collection – PNFC). Remarks to this classification indicate that the operation- al tactical approaches in these complications have their own 9 Klinichna khirurhiia. 2018 November; 85(11) http: //hirurgiya.com.ua Discussion This severe clinical course of the disease was characterized by atypical forms of mesotheliocytes and their nuclei, similar to cells of neoplastic genesis, as observed in 11 (84%) observations vs. 1 (4%) cases (Ȥ2=21.368; p < 0.001). An unbiased assessment of ACP severity is the basis for determining treatment strategy and tactics in patients with this disease. However, the existing integral prognostication scales for disease severity as a general intensive care profile (APACHE II, SAPS, MODS, SOFA), and specifically adapted for acute pancreatitis (Ranson, Leere, Blamey, Balthazar, BISAP) are quite cumbersome and inaccessible to practical application [6, 7]. Recently, studies have been widely conducted to identify certain criteria for disease severity, the so–called ACP predictors, such as the emergence of hydrothorax [8], changes in erythron parameters and iron metabolism [9], neutrophils damage system [10]. The proposed method is affordable, cost– effective and fast, which greatly contributes to addressing this issue. The peculiarities of neutrophil granulocyte changes in the retroperitoneal peripancreatic FC were that both in moderately severe and severe ACP there were cytomorphological damages of cells cytoplasm in the form of degenerative–dystrophic changes. However, in the severe course of disease, in addition to cytoplasm, cell nuclei were also affected by destructive process in the form of karyopyknosis, karyoclasis and karyolysis –5 (71%) observations vs. 1 (7%) case (Ȥ2=10.286; p=0.001). Consequently, the depth of structural changes in mesotheliocytes and neutrophil granulocytes depended on ACP severity. The obtained results provided grounds to work out the “Assessment method of acute necrotic pancreatitis severity” and “Prognostication method for clinical course of acute destructive pancreatitis” [4, 5], according to which the evaluation of cytomorphological changes in mesotheliocytes Results The obtained study data provided grounds to state that as a result of the aggressive environment effect of the peripene- trating fluid collections with high levels of activated enzymes Fig. 1. Degenerative–dystrophic changes in cytoplasm. Castration cell. Fig. 2. Atypically altered mesothelium cells. Anisocytosis, anisonucleosis, polymorphism of cells and nuclei. Fig. 2. Atypically altered mesothelium cells. Anisocytosis, anisonucleosis, polymorphism of cells and nuclei. Fig. 1. Degenerative–dystrophic changes in cytoplasm. Castration cell. Fig. 1. 10 Klinichna khirurhiia. 2018 November; 85(11) http: //hirurgiya.com.ua ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia Fig. 3. Vacuolation of the neutrophil granulocytes cytoplasm. Fig. 4. Changes in granulocyte nuclei. Karyopyknosis, karyoclasis and karyolysis. Fig. 4. Changes in granulocyte nuclei. Karyopyknosis, karyoclasis and karyolysis. Fig. 3. Fig. 3. Fig. 4. Changes in granulocyte nuclei. Karyopyknosis, karyoclasis and karyolysis. Fig. 3. Vacuolation of the neutrophil granulocytes cytoplasm. g 3 Vacuolation of the neutrophil granulocytes cytoplasm. dĂďůĞϭ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŝŶƚƌĂͲĂďĚŽŵŝŶĂů&ŵĞƐŽƚŚĞůŝŽĐǇƚĞƐǁŝƚŚŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚƐĞǀĞƌĞW DŽĚĞƌĂƚĞůǇƐĞǀĞƌĞW ;ŶсϮϰͿ ^ĞǀĞƌĞW ;ŶсϭϯͿ ǇƚŽŵŽƌƉŚŽůŽŐŝĐĂůĐŚĂŶŐĞƐŝŶŵĞƐŽƚŚĞůŝŽĐǇƚĞƐ ĂďƐ͘ й ĂďƐ͘ й DĞƐŽƚŚĞůŝŽĐǇƚĞƐƉƌŽůŝĨĞƌĂƚŝŽŶ ϵ ϯϴ ϭ ϴ ĞŐĞŶĞƌĂƚŝǀĞͲĚǇƐƚƌŽƉŚŝĐĐŚĂŶŐĞƐŝŶĐǇƚŽƉůĂƐŵĂŶĚŵĞƐŽƚŚĞůŝƵŵŶƵĐůĞŝ ϭϰ ϱϴ ϭ ϴ WŽůǇŵŽƌƉŚŝƐŵŽĨŵĞƐŽƚŚĞůŝƵŵĐĞůůƐĂŶĚŶƵĐůĞŝ;ĐĞůůƵůĂƌĂƚǇƉŝĂͿ ϭ ϰ ϭϭ ϴϰ dŽƚĂů͘͘͘ Ϯϰ ϭϬϬ ϭϯ ϭϬϬ dĂďůĞϮ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŶĞƵƚƌŽƉŚŝůŝĐŐƌĂŶƵůŽĐǇƚĞƐŽĨƌĞƚƌŽƉĞƌŝƚŽŶĞĂů&ŝŶŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚ ƐĞǀĞƌĞW DŽĚĞƌĂƚĞůǇƐĞǀĞƌĞW ;ŶсϭϰͿ ^ĞǀĞƌĞW ;ŶсϳͿ ǇƚŽŵŽƌƉŚŽůŽŐŝĐĂůĐŚĂŶŐĞƐŽĨŶĞƵƚƌŽƉŚŝůŐƌĂŶƵůŽĐǇƚĞƐ ĂďƐ͘ й ĂďƐ͘ й ĞŐĞŶĞƌĂƚŝǀĞͲĚǇƐƚƌŽƉŚŝĐĐŚĂŶŐĞƐŝŶĐǇƚŽƉůĂƐŵ ϭϯ ϵϯ Ϯ Ϯϵ ĞƐƚƌƵĐƚŝǀĞĚĂŵĂŐĞƐŽĨŶƵĐůĞƵƐʹŬĂƌǇŽƉǇŬŶŽƐŝƐ͕ŬĂƌǇŽĐůĂƐŝƐĂŶĚŬĂƌǇŽůǇƐŝƐ ϭ ϳ ϱ ϳϭ dŽƚĂů͘͘͘ ϭϰ ϭϬϬ ϳ ϭϬϬ ƌŝƐƚŝĐƐŽĨŝŶƚƌĂͲĂďĚŽŵŝŶĂů&ŵĞƐŽƚŚĞůŝŽĐǇƚĞƐǁŝƚŚŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚƐĞǀĞƌĞW dĂďůĞϭ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŝŶƚƌĂͲĂďĚŽŵŝŶĂů&ŵĞƐŽƚŚĞůŝŽĐǇƚĞƐǁŝƚŚŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚƐĞǀĞƌĞW ŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŝŶƚƌĂͲĂďĚŽŵŝŶĂů&ŵĞƐŽƚŚĞůŝŽĐǇƚĞƐǁŝƚŚŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚƐĞǀĞƌĞW dĂďůĞϭ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŝŶƚƌĂͲĂďĚŽŵŝŶĂů&ŵĞƐŽƚŚĞůŝŽĐǇƚĞƐǁŝƚŚŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚƐĞǀĞƌĞW DŽĚĞƌĂƚĞůǇƐĞǀĞƌĞW ;ŶсϮϰͿ ^ĞǀĞƌĞW ;ŶсϭϯͿ ǇƚŽŵŽƌƉŚŽůŽŐŝĐĂůĐŚĂŶŐĞƐŝŶŵĞƐŽƚŚĞůŝŽĐǇƚĞƐ ĂďƐ͘ й ĂďƐ͘ й DĞƐŽƚŚĞůŝŽĐǇƚĞƐƉƌŽůŝĨĞƌĂƚŝŽŶ ϵ ϯϴ ϭ ϴ ĞŐĞŶĞƌĂƚŝǀĞͲĚǇƐƚƌŽƉŚŝĐĐŚĂŶŐĞƐŝŶĐǇƚŽƉůĂƐŵĂŶĚŵĞƐŽƚŚĞůŝƵŵŶƵĐůĞŝ ϭϰ ϱϴ ϭ ϴ WŽůǇŵŽƌƉŚŝƐŵŽĨŵĞƐŽƚŚĞůŝƵŵĐĞůůƐĂŶĚŶƵĐůĞŝ;ĐĞůůƵůĂƌĂƚǇƉŝĂͿ ϭ ϰ ϭϭ ϴϰ dŽƚĂů͘͘͘ Ϯϰ ϭϬϬ ϭϯ ϭϬϬ dĂďůĞϮ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŶĞƵƚƌŽƉŚŝůŝĐŐƌĂŶƵůŽĐǇƚĞƐŽĨƌĞƚƌŽƉĞƌŝƚŽŶĞĂů&ŝŶŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚ ƐĞǀĞƌĞ W dĂďůĞϮ͘ ǇƚŽůŽŐŝĐĂůĐŚĂƌĂĐƚĞƌŝƐƚŝĐƐŽĨŶĞƵƚƌŽƉŚŝůŝĐŐƌĂŶƵůŽĐǇƚĞƐŽĨƌĞƚƌŽƉĞƌŝƚŽŶĞĂů&ŝŶŵŽĚĞƌĂƚĞůǇƐĞǀĞƌĞĂŶĚ ƐĞǀĞƌĞW DŽĚĞƌĂƚĞůǇƐĞǀĞƌĞW ;ŶсϭϰͿ ^ĞǀĞƌĞW ;ŶсϳͿ ǇƚŽŵŽƌƉŚŽůŽŐŝĐĂůĐŚĂŶŐĞƐŽĨŶĞƵƚƌŽƉŚŝůŐƌĂŶƵůŽĐǇƚĞƐ ĂďƐ͘ й ĂďƐ͘ й ĞŐĞŶĞƌĂƚŝǀĞͲĚǇƐƚƌŽƉŚŝĐĐŚĂŶŐĞƐŝŶĐǇƚŽƉůĂƐŵ ϭϯ ϵϯ Ϯ Ϯϵ ĞƐƚƌƵĐƚŝǀĞĚĂŵĂŐĞƐŽĨŶƵĐůĞƵƐʹŬĂƌǇŽƉǇŬŶŽƐŝƐ͕ŬĂƌǇŽĐůĂƐŝƐĂŶĚŬĂƌǇŽůǇƐŝƐ ϭ ϳ ϱ ϳϭ dŽƚĂů͘͘͘ ϭϰ ϭϬϬ ϳ ϭϬϬ and neutrophil granulocytes in peripancreatic FC content can serve as a commonly available criterion for determining ACP severity. and neutrophil granulocytes in peripancreatic FC content can serve as a commonly available criterion for determining ACP severity. (58%) observations vs. 1 (8%) case (Ȥ2=6.593; p=0.01), while the incidence of mesotheliocytes proliferations did not have statistically significant differences – 9 (38%) vs. 1 (8%) of observations (Ȥ2=2.438; p > 0.05) http: //hirurgiya.com.ua ǖǷȑǹȑȃǹǬ ȁȑǼǿǼǯȑȋ Klinichna khirurhiia and indicative in mesotheliocytes with intra–abdominal FC localization and neutrophil granulocytes with their retroperitoneal arrangement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p g 2. Assessment of the extent of identified cytomorphological damage in cellular composition of FC sediment can serve as an objective and publicly available criterion of ACP severety. 8NUDLQD03.1]DLDYQ\N\WDSDWHQWRYODVQ\N\.YLW >,Q8NUDLQLDQ@ 1\FK\WDLOR0<X$QGULXVKFKHQNR'90DKORYDQ\L9$$QGULXVKFKHQ NR93.KDUDNWHU\VW\N\VXFKDVQRKREDNWHULLQRKRFK\QQ\NDSU\KRV WURPXSDQNUHDW\WLWD\LNKNOLQLFKQH]QDFKHQQLDGOLDREJUXQWXYDQQLDOL NXYDOQRLWDNW\N\0H]KGXQDURGQ\L]KXUQDODQW\E\RW\N\\SURE\RW\N\ ± Conclusions 1. Clinical course of ACP shows cytomorphological changes in peripancreatic FC contents, which are the most significant 11 Klinichna khirurhiia. 2018 November; 85(11) http: //hirurgiya.com.ua http: //hirurgiya.com.ua Klinichna khirurhiia. 2018 November; 85(11) References %DQNV3$%ROOHQ7/'HUYHQLV&RRV]HQ+-RKQVRQ&'6DUU0 HWDO$FXWH3DQFUHDWLWLV&ODVVL¿FDWLRQ:RUNLQJURXS&ODVVL¿FDWLRQ RIDFXWHSDQFUHDWLWLV±UHYLVLRQRIWKH$WODQWDFODVVL¿FDWLRQDQG GH¿QLWLRQVE\LQWHUQDWLRQDOFRQVHQVXVXW±GRL JXWMQO±± %ROOHQ7/6LQJK9.0DXUHU55HSDV.YDQ(V+:%DQNV3$HWDO $FRPSDUDWLYHHYDOXDWLRQRIUDGLRORJLFDQGFOLQLFDOVFRULQJV\VWHPVLQ WKHHDUO\SUHGLFWLRQRIVHYHULW\LQDFXWHSDQFUHDWLWLV$P-DVWURHQWHU RO$SU±GRLDMJ :RUNLQJURXS,$3$3$$FXWH3DQFUHDWLWLVXLGHOLQHV,$3$3$HY LGHQFH±EDVHGJXLGHOLQHVIRUWKHPDQDJHPHQWRIDFXWHSDQFUHDWLWLV 3DQFUHDWRORJ\-XO±$XJ6XSSOH±GRLM SDQ 5XV\Q9,)LOLS66+LGURWRUDNF±SUHG\NWRUYD]KNRVWLSURWLNDQQLDKRV WURKRSDQNUHDW\WX8NUDLQVN\L]KXUQDONKLUXUKLL±>,Q 8NUDLQLDQ@ +RGOHYVN\L$,6DYROLXN6,0HOQ\N723URKQRVW\FKQH]QDFKHQQLD SRND]Q\NLYV\VWHP\HU\WURQXWDREPLQX]DOL]DXNKYRU\NKQDKRVWU\L SDQNUHDW\W=ELUQ\NQDXNRY\NKURELW;;,,,]ƍL]GXNKLUXUKLY8NUDLQ\ .\LY±>,Q8NUDLQLDQ@ $LWNHQ(/RXJK9-RQH$2EVHUYDWLRQVWXG\RILQWUD±DEGRPLQDOSUHV VXUHPRQLWRULQJLQDFXWHSDQFUHDWLWLV6XUJHU\0D\± GRLMVXUJ $QGULXVKFKHQNR9Ɋ.RKXW/0$QGUXVKHYVND2<XY\QDNKLGQ\N\ 'DQ\OR+DO\WVN\/YLY1DWLRQDO0HGLFDO8QLYHUVLW\SDWHQWRYODVQ\N 6SRVLERWVLQN\WLD]KNRVWLKRVWURKRQHNURW\FKQRKRSDQNUHDW\WX3DWHQW 8NUDLQ\1R/LVW>,Q8NUDLQLDQ@ +RPROLDNR,97XPDVRYD.3.ORFKNRYD1<H,QGHNVSRVKNRG]KHQ QLDV\VWHP\QHLWUR¿OLY\DNIDNWRUSURKQR]XNKLUXUKLFKQ\NKXVNODGQHQ .OLQNKLU6XSSO±>,Q8NUDLQLDQ@ 12 Klinichna khirurhiia. 2018 November; 85(11)	49,059
https://openalex.org/W2401939449_2	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	1,515	2,793	Algunos entrevistados mencionaron: Este tipo de turismo nos afecta a todos los transportistas… ya le incluye todo para ir a pescar, la casa y toda la transportación… eso es lo que nos afecta… los americanos que se quedaron acá… uno dice: no está mal que se queden, son libres de hacerlo, pero ya que vengan y hagan sus negocios acá, eso ya está mal… están viendo que casi no hay turismo y el poco que pudiera llegar se lo jinetean… cómo voy a creer… échese una vuelta al aeropuerto, se va a cerciorar que mucha gente de aquí americana, dizque van por sus “amigos”… ¡No, que es mi amigo… Vengo por él! [lo expresa imitando a un turista estadounidense], pero ya le está dando el servicio de hotel, el servicio de barco, ya le está dando el servicio de transportación, eso es lo que nos afecta a nosotros los taxistas [Michael, taxista]. Los servicios de luz y agua escasean en temporada alta. Al respecto, un entrevistado comentó: Cuando es la temporada alta, que es de octubre a mayo, sí disminuye el abasto de agua… yo creo que porque somos más con la llegada de los turistas y no alcanza para todos… pero pues yo no creo que se la quiten a los hoteles o a los condominios de los gringos [turista estadounidense] la verdad… y con la luz lo mismo [Aldo, comerciante]. En cuanto a los costos de los turistas estadounidenses con casa en la comunidad, los encuestados distinguen que representan competencia desleal para los prestadores de servicios turísticos locales (66 % está totalmente de acuerdo o de acuerdo, 8.8 % ni de acuerdo ni en desacuerdo y 24.9 % en desacuerdo o totalmente en desacuerdo). Percepción del turismo Para dar más profundidad a la investigación, la percepción de los residentes sobre la actividad turística también fue explorada en el trabajo de campo cualitativo. La mayoría de los entrevistados se muestra favorable a la actividad turística, muchos señalaron esperar mejores momentos y oportunidades con el turismo en Loreto. 142 Qué bueno que el turismo mantenga a Loreto… no sé qué sería sin él… el futuro de Loreto se encuentra en el turismo… vienen tiempos muy buenos para Loreto, ya Mendoza Ontiveros, González Sosa Teoría y Praxis 16 (2014: 117-146) hay más publicidad… la gente empieza a venir más y si llegaran a venir mucho más, eso va a ser muy bueno para todos [José, gerente de hotel]. Está bien que vengan los turistas, nos dan trabajo, si vinieran más estaría mucho mejor la situación… ahorita está muy bajo el turismo pero ya mejorará… es lo mejor para Loreto [Mario, taxista]. Yo sí me beneficio del turismo, está muy bien que vengan a gastar y ayudar a la economía de la comunidad… está muy, muy bien [Aldo, comerciante]. Yo sí quisiera que fueran más los turistas que nos visitan, pero el poco turismo que viene es bien recibido [Raquel, cocinera]. Consideraciones finales Los resultados revelan que los residentes locales apoyan el turismo ya que se aprecian más beneficiados que afectados (en cuanto a empleos, infraestructura, inversión, entre otros aspectos), por ello se advierte una actitud favorable o positiva hacia el turismo y están deseosos de que Loreto siga creciendo para recibir más turismo. Sin embargo, algunos residentes creen que los costos del turismo prevalecen sobre los beneficios (problemas sociales y ambientales, cambio en el estilo de vida, empleos mal pagados, entre otros). De esta forma, desde la teoría del intercambio social se evidencia que estas personas tendieron a evaluar los impactos de forma negativa. Los residentes están dispuestos a participar en tal intercambio si creen que es probable obtener beneficios sin incurrir en costos inaceptables (drogadicción, prostitución, alcoholismo, competencia desleal por parte de turistas residenciales, entre otros). Así, quienes perciben que los beneficios son mayores que los costos están dispuestos a participar en el intercambio y, por lo tanto, apoyan el desarrollo del turismo local futuro porque de ello viven u obtienen su sustento. Asimismo, no se perciben efectos como la generación de basura, el incremento en el tráfico vehicular, aglomeraciones en espacios públicos o aquellos impactos que han emergido en destinos con una mayor afluencia de turistas. En general, la comunidad local del cip Loreto observa que el turismo les trae más beneficios, sobre todo en lo económico y en las obras y programas 143 Impactos socioculturales del turismo en el Centro Integralmente Planeado Loreto. sociales y ambientales que realizan los turistas americanos con casa en la comunidad (que pretenden mejorar el lugar donde viven algunos meses del año) o las personas jubiladas que buscan un buen sitio para pasar sus últimos años. Estos turistas hacen campañas de limpieza en las playas, así como donativos a escuelas y centros de salud e impulsaron el uso del cinturón de seguridad en los automóviles para evitar muertes en accidentes. Cabe mencionar que la percepción es un acto subjetivo que puede no corresponder con la realidad objetiva, pues se percibe desde los marcos de referencia, la historia personal y la experiencia de vida; no obstante, esto no puede ser descalificado o ignorado, ya que a partir de la percepción se toman posturas o se responde a una situación determinada. De ahí la relevancia de conocer el punto de vista de la comunidad; identificar los beneficios y los costos que le atribuyen al turismo puede dar una idea de qué está viendo la gente para predecir el rumbo de sus decisiones y comportamiento. Como hipótesis puede postularse que estos resultados se deben en gran medida a que el turismo en el cip Loreto no se ha masificado como por ejemplo en Cancún y por el perfil del turista que los visita, pues van más para estar en contacto con la naturaleza haciendo actividades como la pesca deportiva o la observación de aves, ballenas o delfines, por lo que aún sienten más beneficios que problemas. Por otra parte, se puede inferir que la percepción de la comunidad local sobre los costos y beneficios del turismo (impactos) no es homogénea, existen variados puntos de vista. Durante los trabajos de campo se comprobó que los residentes que tienen un beneficio directo (económico), como los meseros, restauranteros, recepcionistas, hoteleros, entre otros, tendieron a evaluar positivamente los impactos del turismo: aquellos que se encontraban lejos de las áreas con actividad turística se mostraron neutrales en su percepción; y aquellos afectados por los costos del turismo (económico y social), como taxistas y pescadores, principalmente, hacían evidente su desaprobación hacia el turismo y sus impactos. Fuentes consultadas 144 Coll, C. et al. (1992). Enseñanza y aprendizaje de conceptos, procedimientos y Mendoza Ontiveros, González Sosa Teoría y Praxis 16 (2014: 117-146) actitudes. México: Santillana. Cooper, Ch. et al. (2007). Turismo, teoría y práctica. Madrid: Síntesis. Doğan, H. 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https://openalex.org/W3170053175	OpenAlex	Open Science	CC-By	2,021	Multivariate Normal Approximation on the Wiener Space: New Bounds in the Convex Distance	Ivan Nourdin	English	Spoken	7,689	16,482	Abstract We establish explicit bounds on the convex distance between the distribution of a vector of smooth functionals of a Gaussian ﬁeld, and that of a normal vector with a positive deﬁnite covariance matrix. Our bounds are commensurate to the ones obtained by Nourdin, Peccati and Réveillac (2010) for the (smoother) 1-Wasserstein distance, and do not involve any addi- tional logarithmic factor. One of the main tools exploited in our work is a recursive estimate on the convex distance recently obtained by Schulte and Yukich (2019). We illustrate our abstract results in two diﬀerent situations: (i) we prove a quantitative multivariate fourth moment theorem for vectors of multiple Wiener-Itô integrals, and (ii) we characterise the rate of convergence for the ﬁnite-dimensional distributions in the functional Breuer-Major theorem. arXiv:2001.02188v2 [math.PR] 25 Feb arXiv:2001.02188v2 [math.PR Keywords: Breuer-Major Theorem; Convex Distance; Fourth Moment Theorems; Gaussian Fields; Malliavin-Stein Method; Multidimensional Normal Approximations. AMS 2020 Classiﬁcation: 60F05; 60G15; 60H07 Multivariate normal approximation on the Wiener space: new bounds in the convex distance Ivan Nourdin∗, Giovanni Peccati†and Xiaochuan Yang‡ February 26, 2021 February 26, 2021 2001.02188v2 [math.PR] 25 Feb 2021 ∗University of Luxembourg, Department of Mathematics, inourdin@gmail.com †University of Luxembourg, Department of Mathematics, giovanni.peccati@gmail.com ‡ University of Bath, Department of Mathematical Sciences, xiaochuan.j.yang@gmail.com 1For instance, one has that dc(TF, TG) = dc(F, G), whenever the mapping T : Rm −→Rm is an invertible aﬃne mapping — see e.g. [4, 28] for more details. 1 Introduction In Appendix A we will also implicitly use the well-known representation dW (F, NΣ) = inf (U,V) E ∥U −V∥Rm , where the inﬁmum runs over all couplings (U, V) of F and NΣ. See [30, Ch. I-6] for a proof of this fact, and for further relevant properties of Wasserstein distances. where the inﬁmum runs over all couplings (U, V) of F and NΣ. See [30, Ch. I-6] for a proof of this fact, and for further relevant properties of Wasserstein distances. The main contributions of our paper are described in full detail in Section 1.4 and Section 1.5. Section 1.1 contains some elements of Malliavin calculus that are necessary in order to state our ﬁndings. Section 1.2 discusses some estimates on the smooth distance d2 (to be deﬁned therein) that can be obtained by interpolation techniques, whereas Section 1.3 provides an overview of the main results of [19]. Remark on notation. From now on, every random element is assumed to be deﬁned on a common probability space (Ω, F, P), with E denoting expectation with respect to P. For p ≥1, we write Lp(Ω) := Lp(Ω, F, P). 1 Introduction Fix m ≥1, and consider random vectors F and G with values in Rm. The convex distance between the distributions of F and G is deﬁned as dc(F, G) := sup h∈Im Eh(F) −Eh(G) , (1) (1) where the supremum runs over the class Im of indicator functions of the measurable convex subsets of Rm. For m ≥2, the distance dc represents a natural counterpart to the well-known Kolmogorov distance on the class of probability distributions on the real line, and enjoys a number of desirable invariance properties that make it well-adapted to applications1. The aim of the present note is to establish explicit bounds on the quantity dc(F, G), in the special case where F is a vector of smooth functionals of an inﬁnite-dimensional Gaussian ﬁeld, and G = NΣ is a m-dimensional centered Gaussian vector with covariance Σ > 0. Our main tool is the so-called Malliavin-Stein method for probabilistic approximations [17], that we 1 will combine with some powerful recursive estimates on dc, recently derived in [29] in the context of multidimensional second-order Poincaré inequalities on the Poisson space — see Lemma 2.1 below. Multidimensional normal approximations in the convex distance have been the object of an intense study since several decades, mostly in connection with multivariate central limit theorems (CLTs) for sums of independent random vectors — see e.g. [4, 10, 13, 14] for some classical references, as well as [28] for recent advances and for a discussion of further relevant literature. The speciﬁc challenge we are setting ourselves in the present work is to establish bounds on the quantity dc(F, NΣ) that coincide (up to an absolute multiplicative constant) with the bounds deduced in [19] on the 1-Wasserstein distance dW (F, NΣ) := sup h∈Lip(1) \|Eh(F) −Eh(NΣ)\| , (2) (2) where Lip(1) denotes the class of C1 mappings h : Rm →R with Lipschitz constant not exceeding 1. We will see that our estimates systematically improve the bounds that one can infer from the general inequality q dc(F, NΣ) ≤K q dW (F, NΣ), (3) (3) where K is an absolute constant uniquely depending on Σ. For the sake of completeness, a full proof of (3) is presented in Appendix A, where one can also ﬁnd more details on the constant K. Remark 1.1. In order for the quantity dW (F, NΣ) to be well-deﬁned, one needs that E∥F∥Rm < ∞. 1.1 Elements of Malliavin calculus The reader is referred e.g. to the monographs [17, 23, 24] for a detailed discussion of the concepts and results presented in this subsection. Let H be a real separable Hilbert space, and write ⟨·, ·⟩H for the corresponding inner product. In what follows, we will denote by X = {X(h) : h ∈H} an isonormal Gaussian process over H, that is, X is a centered Gaussian family indexed by the elements of H and such that E[X(h)X(g)] = ⟨h, g⟩H for every h, g ∈H. For the rest of the paper, we will assume without loss of generality that F coincides with the σ-ﬁeld generated by X. 2 2 Every F ∈L2(Ω) admits a Wiener-Itô chaos expansion of the form Every F ∈L2(Ω) admits a Wiener-Itô chaos expansion of the form F = EF + ∞ X q=1 Iq(fq), F = EF + ∞ X q=1 Iq(fq), where fq belongs to the symmetric qth tensor product H⊙q (and is uniquely determined by F), and Iq(fq) is the q-th multiple Wiener-Itô integral of fq with respect to X. One writes F ∈D1,2(Ω) if X q≥1 qq! ∥fq∥2 H⊗q < ∞. For F ∈D1,2(Ω), we denote by DF the Malliavin derivative of F, and recall that DF is by deﬁnition a random element with values in H. The operator D satisﬁes a fundamental chain rule: if ϕ : Rm →R is C1 and has bounded derivatives and if F1, . . . , Fm ∈D1,2(Ω), then ϕ(F1, . . . , Fm) ∈D1,2(Ω) and Dϕ(F1, . . . , Fm) = m X i=1 ∂iϕ(F1, . . . , Fm)DFi. (4) (4) For general p > 2, one writes F ∈D1,p(Ω) if F ∈Lp(Ω) ∩D1,2(Ω) and E[∥DF∥p H] < ∞. For general p > 2, one writes F ∈D1,p(Ω) if F ∈Lp(Ω) ∩D1,2(Ω) and E[∥DF∥p H] < ∞. The adjoint of D, customarily called the divergence operator or the Skorohod integral, is denoted by δ and satisﬁes the duality formula, The adjoint of D, customarily called the divergence operator or the Skorohod integral, is denoted by δ and satisﬁes the duality formula, E[δ(u)F] = E[⟨u, DF⟩H] (5) (5) for all F ∈D1,2(Ω), whenever u : Ω→H is in the domain Dom(δ) of δ. for all F ∈D1,2(Ω), whenever u : Ω→H is in the domain Dom(δ) of δ. 1.1 Elements of Malliavin calculus The generator of the Ornstein-Uhlenbeck semigroup, written L, is deﬁned by the rela- tion LF = −P q≥1 qIq(fq) for every F such that P q≥1 q2q! ∥fq∥2 H⊗q < ∞. The pseudo-inverse of L, denoted by L−1, is the operator deﬁned for any F ∈L2(Ω) as L−1F = −P q≥1 1 qIq(fq). The crucial relation that links the objects introduced above is the identity F = EF −δ(DL−1F), (6) (6) which is valid for any F ∈L2(Ω) (in particular, one has that, for every F ∈L2(Ω), DL−1F ∈ Dom(δ)). It is then immediate that d2(F, NΣ) ≤1 2 E∥MF −Σ∥H.S.. (9) (9) Inequalities in the spirit of (9) were derived e.g. in [18] (in the context of limit theorems for homogeneous sums) and [27] (in the framework of multivariate normal approximations on the Poisson space) — see also [29] and the references therein. 1.2 Bounds on the smooth distance d2 Fix m ≥1 and assume that F = (F1, ..., Fm) is a centered random vector in Rm whose compo- nents belong to D1,2(Ω). Without loss of generality, we can assume that each Fi is of the form Fi = δ(ui) for some ui ∈Dom(δ); indeed, by virtue of (6) one can always set ui = −DL−1Fi (although this is by no means the only possible choice). Let also NΣ = (N1, . . . , Nm) be a centered Gaussian vector with invertible m × m covariance matrix Σ = {Σ(i, j)} = {Σ(i, j) : i, j = 1, ..., m}. Finally, consider the so-called d2 distance (between the distributions of F and NΣ) deﬁned by d2(F, NΣ) = sup h Eh(F) −Eh(NΣ) , d2(F, NΣ) = sup h Eh(F) −Eh(NΣ) , where the supremum is taken over all C2 functions h : Rm →R that are 1-Lipschitz and such that supx∈Rm ∥(Hess h)(x)∥H.S. ≤1; here, Hess h stands for the Hessian matrix of h, whereas where the supremum is taken over all C2 functions h : Rm →R that are 1-Lipschitz and such that supx∈Rm ∥(Hess h)(x)∥H.S. ≤1; here, Hess h stands for the Hessian matrix of h, whereas 3 3 ∥· ∥H.S. (resp. ⟨·, ·⟩H.S.) denotes the Hilbert-Schmidt norm (resp. scalar product), that is, ⟨A, B⟩H.S. = Tr(ABT) = P 1≤i,j≤m A(i, j)B(i, j) and ∥A∥2 H.S. = ⟨A, A⟩H.S. for any m×m matrices A = {A(i, j)} and B = {B(i, j)}. ∥· ∥H.S. (resp. ⟨·, ·⟩H.S.) denotes the Hilbert-Schmidt norm (resp. scalar product), that is, ⟨A, B⟩H.S. = Tr(ABT) = P 1≤i,j≤m A(i, j)B(i, j) and ∥A∥2 H.S. = ⟨A, A⟩H.S. for any m×m matrices A = {A(i, j)} and B = {B(i, j)}. For a given h : Rm →R ∈C2 with bounded partial derivatives, let us introduce its molliﬁ- cation at level √ t, deﬁned by ht(x) := E[h( √ tNΣ + √ 1 −tx)], x ∈Rm. (7) (7) (7) One has One has Eh(NΣ) −Eh(F) = Z 1 0 d dtE[ht(F)]dt = 1 2 m X i=1 Z 1 0 1 √ tE[∂ih( √ tNΣ + √ 1 −tF)Ni] − 1 √1 −tE[∂ih( √ tNΣ + √ 1 −tF)Fi] dt. where MF is the random m × m matrix given by where MF is the random m × m matrix given by MF (i, j) = ⟨DFi, uj⟩H. (8) MF (i, j) = ⟨DFi, uj⟩H. (8) It is then immediate that 1.2 Bounds on the smooth distance d2 Supposing in addition (and without loss of generality) that F and NΣ are independent, we can write, using the Gaussian integration by parts Supposing in addition (and without loss of generality) that F and NΣ are independent, we can write, using the Gaussian integration by parts E[∂ih( √ tNΣ + √ 1 −tF)Ni] = EFENΣ[∂ih( √ tNΣ + √ 1 −tF)Ni] = √ t m X j=1 E[∂2 ijh( √ tNΣ + √ 1 −tF)]E[NiNj], and, combining the duality formula (5) with the chain rule (4), and, combining the duality formula (5) with the chain rule (4), and, combining the duality formula (5) with the chain rule (4), E[∂ih( √ tNΣ + √ 1 −tF)Fi] = ENΣEF[∂ih( √ tNΣ + √ 1 −tF)Fi] = √ 1 −t m X j=1 E[∂2 ijh( √ tNΣ + √ 1 −tF)⟨DFi, uj⟩H]. Putting everything together leads to Putting everything together leads to Eh(NΣ) −Eh(F) = 1 2 Z 1 0 E[⟨(Hess h)( √ tNΣ + √ 1 −tF), Σ −MF ⟩H.S.]dt, 1.3 Bounds on the 1-Wasserstein distance ≤√m ∥Σ−1∥op ∥Σ∥1/2 op , (13) (13) and ∥·∥op is the operator norm for m × m matrices. The estimate (12) is the main result of [19] (see also [17, Theorem 6.1.1]), whereas a self-contained proof of (13) can be found in [17, Proposition 4.3.2]. and ∥·∥op is the operator norm for m × m matrices. The estimate (12) is the main result of [19] (see also [17, Theorem 6.1.1]), whereas a self-contained proof of (13) can be found in [17, Proposition 4.3.2]. 1.3 Bounds on the 1-Wasserstein distance For random vectors F and NΣ as in the previous section, we will now discuss a suitable method for assessing the quantity dW (F, NΣ) deﬁned in (2), that is, for uniformly bounding the absolute diﬀerence \|Eh(F) −Eh(NΣ)\| over all 1-Lipschitz functions h of class C1. 4 Since we do not assume h to be twice diﬀerentiable, the method presented in Section 1.2 no longer works. A preferable approach is consequently the so-called ‘Malliavin-Stein method’, introduced in [16] in dimension 1, and later extended to the multivariate setting in [19]. Let us brieﬂy recall how this works (see [17, Chapter 4 and Chapter 6] for a full discussion, and [1] for a constantly updated list of references). Start by considering the following Stein’s equation, with h : Rm →R given and f : Rm → R unknown: m X i,j=1 Σ(i, j)∂2 ijf(x) − m X i=1 xi∂if(x) = h(x) −Eh(NΣ), x ∈Rm. (10) (10) When h ∈C1 has bounded partial derivatives, it turns out that (10) admits a solution f = fh of class C2 and whose second partial derivatives are bounded — see e.g. [17, Proposition 4.3.2] for a precise statement. Taking expectation with respect to the distribution of F in (10) gives When h ∈C1 has bounded partial derivatives, it turns out that (10) admits a solution f = fh of class C2 and whose second partial derivatives are bounded — see e.g. [17, Proposition 4.3.2] for a precise statement. Taking expectation with respect to the distribution of F in (10) gives Eh(F) −Eh(NΣ) = m X i,j=1 Σ(i, j)E[∂2 ijfh(F)] − m X i=1 E[Fi∂ifh(F)]. We can apply again the duality formula (5) together with the chain rule (4), to deduce We can apply again the duality formula (5) together with the chain rule (4), to deduce that the duality formula (5) together with the chain rule (4), to deduce that Eh(F) −Eh(NΣ) = E[⟨(Hess fh)(F), MF −Σ⟩H.S.], (11) (11) where MF is deﬁned in (8). Taking the supremum over the set of all 1-Lipschitz functions h : Rm →R of class C1, we infer where MF is deﬁned in (8). Taking the supremum over the set of all 1-Lipschitz functions h : Rm →R of class C1, we infer dW (F, NΣ) ≤c1 E∥MF −Σ∥H.S., (12) (12) with c1 = sup h∈Lip(1) sup x∈Rm ∥(Hess fh)(x)∥H.S. 1.4 Main results: bounds on the convex distance The principal aim of the present paper is to address the following natural question: can one obtain a bound similar to (12) for distances based on non-smooth test functions h : Rm →R, such as e.g. indicator functions of measurable convex subsets of Rm? If h is such an indicator function, then we recall e.g. from [29, Lemma 2.2] that, for all t ∈(0, 1), Eh(F) −Eh(NΣ) ≤4 3\|Eht(F) −Eht(NΣ)\| + 20m √ 2 √ t 1 −t, where ht stands for the molliﬁcation at level √ t of h, as deﬁned in (7). Let ft = fht be the solution of the Stein’s equation (10) associated with h = ht. In [6] (see also [29]), it is shown that where ht stands for the molliﬁcation at level √ t of h, as deﬁned in (7). Let ft = fht be the solution of the Stein’s equation (10) associated with h = ht. In [6] (see also [29]), it is shown that max 1≤i,j≤m sup x∈Rm \|∂2 ijft(x)\| ≤c2\| log t\|, max 1≤i,j≤m sup x∈Rm \|∂2 ijft(x)\| ≤c2\| log t\|, 5 5 with c2 = c2(m, Σ) a constant depending only on m and Σ. Combining such an estimate with (11) yields the existence of a constant c3 = c3(m, Σ) > 0 such that Eh(F) −Eh(NΣ) ≤c3 E∥MF −Σ∥H.S.\| log t\| + √ t 1 −t ! . (14) (14) From (14), it is straightforward to deduce the existence of c4 = c4(m, Σ) > 0 such that From (14), it is straightforward to deduce the existence of c4 = c4(m, Σ) > 0 such that dc(F, NΣ) ≤c4 E∥MF −Σ∥H.S. log{E∥MF −Σ∥H.S.} . (15) (15) Comparing (15) with (9) and (12) shows that such a strategy yields a bound on dc(F, NΣ) diﬀer- ing from those deduced above for the distances d2 and dW by an additional logarithmic factor. See also [11, 20] for more inequalities analogous to (15) — that is, displaying a multiplicative logarithmic factor — related respectively to the (multivariate) Kolmogorov and total variation distances. In this paper, we will show that one can actually remove the redundant logarithmic factor on the right-hand side of (15), thus yielding a bound on dc(F, NΣ) that is commensurate to (9) and (12) (with moreover an explicit multiplicative constant). Our main result is the following: Theorem 1.2. Let F = (F1, ..., Fm) = (δ(u1), . . . 1.4 Main results: bounds on the convex distance , δ(um)) be a vector in Rm of centered random variables such that ui ∈Dom(δ), for i = 1, ..., m. Let also NΣ = (N1, . . . , Nm) be a centered Gaussian vector with invertible m × m covariance matrix Σ = {Σ(i, j)}. Then dc(F, NΣ) ≤402 Σ−1 3/2 op + 1 m41/24 q E ∥MF −Σ∥2 H.S. , with MF deﬁned in (8). As anticipated, to prove Theorem 1.2, we shall combine the somewhat classical smoothing estimate (14) with a remarkable bound by Schulte and Yukich [29]. As anticipated, to prove Theorem 1.2, we shall combine the somewhat classical smoothing estimate (14) with a remarkable bound by Schulte and Yukich [29]. 1.5 Applications We illustrate the use of Theorem 1.2 by developing two examples in full detail. Quantitative fourth moment theorems. A fourth moment theorem (FMT) is a mathemat- ical statement implying that a given sequence of centered and normalized random variables converges in distribution to a Gaussian limit, as soon as the corresponding sequence of fourth moments converges to 3 (that is, to the fourth moment of the standard Gaussian distribution). Distinguished examples of FMTs are e.g. de Jong’s theorem for degenerate U-statistics (see [8, 9]) as well as the CLTs for multiple Wiener-Itô integrals proved in [25, 26]; the reader is referred to the webpage [1] for a list (composed of several hundreds of papers) of applications and extensions of such results, as well as to the lecture notes [31] for a modern discussion of their relevance in mathematical physics. Our ﬁrst application of Theorem 1.2 is a quantitative multivariate fourth moment theorem for a vector of multiple Wiener-Itô integrals, considerably extending the qualitative multivariate results proved in [26]. Note that such a result was already obtained by Nourdin and Rosiński [22, Theorem 4.3] for the 1-Wasserstein distance dW . Thanks to Theorem 1.2, it is not diﬃcult to generalize their result to the dc metric. 6 Corollary 1.3. Fix m ≥1 as well as q1, . . . , qm ≥1. Let F = (F1, ..., Fm) where Fi = Iqi(fqi) with fqi ∈H⊙qi. Let NΣ be a centered Gaussian vector with covariance matrix Σ = (EFiFj)i,j∈[m] supposed to be invertible. Then dc(F, NΣ) ≤402 Σ−1 3/2 op + 1 m41/24 q E ∥F∥4 −E ∥NΣ∥4. In particular, for a vector F of multiple Wiener-Itô integrals to be close in the convex distance to a centered Gaussian vector NΣ with matching covariance matrix, it is enough that E∥F∥4 ≈E ∥NΣ∥4. In particular, for a vector F of multiple Wiener-Itô integrals to be close in the convex distance to a centered Gaussian vector NΣ with matching covariance matrix, it is enough that E∥F∥4 ≈E ∥NΣ∥4. The multivariate Breuer-Major theorem. The second example concerns the convergence towards a Brownian motion occurring in the Breuer-Major theorem proved in [5]. Let us brieﬂy recall this fundamental result (see [17, Chapter 7] for an introduction to the subject, as well as [15, 7] for recent advances in a functional setting). 1.5 Applications 7 7 Therefore, choosing A appropriately, it suﬃces to consider the vector Fn = (Fn,1, ..., Fn,m) with Fn,i = 1 √n ⌊nti⌋ X k=⌊nti−1⌋+1 ϕ(Gk), i ∈[m] and obtain the rate of convergence for Fn d −→N(0, σ2Diag(t1 −t0, ..., tm −tm−1)) =: NΣ. (17) (17) The following result provides a quantitative version of this CLT with respect to the distance dc. Recall from [21] that the minimal regularity assumption over ϕ for obtaining rates of convergence via the Malliavin-Stein method is that ϕ ∈D1,4(R, γ), meaning that ϕ is absolutely continuous and both ϕ and its derivative ϕ′ belong to L4(R, γ). We say that ϕ is 2-sparse if its expansion in Hermite polynomials does not have consecutive non-zero coeﬃcients. In particular, even functions are 2-sparse. The following result provides a quantitative version of this CLT with respect to the distance dc. Recall from [21] that the minimal regularity assumption over ϕ for obtaining rates of convergence via the Malliavin-Stein method is that ϕ ∈D1,4(R, γ), meaning that ϕ is absolutely continuous and both ϕ and its derivative ϕ′ belong to L4(R, γ). We say that ϕ is 2-sparse if its expansion in Hermite polynomials does not have consecutive non-zero coeﬃcients. In particular, even functions are 2-sparse. Corollary 1.4. Let Fn and NΣ be given in (17). Suppose that ϕ ∈D1,4(R, γ) with Hermite rank d ≥1. Then, i) There exists a constant C depending only on ϕ, m, Σ such that for each n ∈N, dc(Fn, NΣ) ≤C m X i,j=1 \|Σ(i, j) −E[FiFj]\| + Cn−1 2  X \|k\|<n \|ρ(k)\|   3 2 . ii) If d = 2, ϕ is 2-sparse and b ∈[1, 2], then there exists a constant C depending only on ϕ, m, Σ such that for each n ∈N, ii) If d = 2, ϕ is 2-sparse and b ∈[1, 2], then there exists a constant C depending only on ϕ, m, Σ such that for each n ∈N, ii) If d = 2, ϕ is 2-sparse and b ∈[1, 2], then there exists a constant C depending only on ϕ, m, Σ such that for each n ∈N, dc(Fn, NΣ) ≤C m X i,j=1 \|Σ(i, j) −E[FiFj]\| + Cn−( 1 b −1 2 )  X \|k\|<n \|ρ(k)\|2   1 2  X \|k\|<n \|ρ(k)\|b   1 b . 1.5 Applications iii) If d = 2, ϕ is 2-sparse, and P k∈Z \|ρ(k)\|2 < ∞, then as n →∞, dc(Fn, NΣ) →0. dc(Fn, NΣ) →0. The rest of the note is organized as follows. The proof of Theorem 1.2 is given in Section 2.1, Corollary 1.3 in Section 2.2, Corollary 1.4 in Section 2.3. We use C to denote a generic constant whose value may change from line to line. 1.5 Applications Let {Gk : k ∈Z} be a centered Gaussian stationary sequence with ρ(j −k) = E[GjGk] and ρ(0) = 1; in particular, Gk ∼N(0, 1) for all k. Let ϕ ∈L2(R, γ) where γ(dx) = (2π)−1/2e−x2/2dx denotes the standard Gaussian measure on R. Since the Hermite polynomials {Hk : k ≥0} form an orthonormal basis of L2(R, γ), one has ϕ = X k≥d akHk, with d ∈N and ad ̸= 0. The index d is known as the Hermite rank of ϕ ∈L2(R, γ). Suppose in addition that R R ϕdγ = E[ϕ(G0)] = 0, that is, suppose d ≥1. The Breuer-Major theorem [5] states the following: if P k∈Z \|ρ(k)\|d < ∞, then    1 √n ⌊nt⌋ X k=1 ϕ(Gk) : t ≥0    f.d.d. −→{σW(t) : t ≥0} (16) (16) where W is a standard Brownian motion, f.d.d. −→indicates convergence in the sense of ﬁnite- dimensional distributions, and where W is a standard Brownian motion, f.d.d. −→indicates convergence in the sense of ﬁnite- dimensional distributions, and σ2 := X k≥d a2 kk! X j∈Z ρ(j)k ∈[0, ∞), (That σ2 is a well-deﬁned positive real number is part of the conclusion.) We refer to our note [21] and references therein for results on the rate of convergence in the total variation distance for one-dimensional marginal distributions (that is, in dimension 1). We intend to apply Theorem 1.2 to address the rate of convergence for the following multivariate CLT implied by (16): for every 0 = t0 < t1 < ... < tm = T < ∞,  1 √n ⌊nt1⌋ X k=1 ϕ(Gk), ..., 1 √n ⌊ntm⌋ X k=1 ϕ(Gk)   d −→N(0, Σ(t1, ..., tm)) where d −→indicates converges in distribution, and N(0, Σ(t1, ..., tm)) is a m-dimensional centred Gaussian vector with covariance Σ(t1, ..., tm) having entries σ2ti ∧tj, i, j = 1, ..., m. Notice that for any m × m invertible matrix A, where d −→indicates converges in distribution, and N(0, Σ(t1, ..., tm)) is a m-dimensional centred Gaussian vector with covariance Σ(t1, ..., tm) having entries σ2ti ∧tj, i, j = 1, ..., m. Notice that for any m × m invertible matrix A, dc(F, G) = dc(AF, AG). 1.6 Acknowledgments We thank Simon Campese and Nicola Turchi for pointing out an error in an earlier version. I. Nourdin is supported by the FNR grant APOGee (R-AGR-3585-10) at Luxembourg University; G. Peccati is supported by the FNR grant FoRGES (R-AGR-3376-10) at Luxembourg University; X. Yang was supported by the FNR Grant MISSILe (R-AGR-3410-12-Z) at Luxembourg and Singapore Universities. 8 8 2.1 Proof of Theorem 1.2 We divide the proof into several steps. Step 1 (smoothing). For any bounded and measurable h and t ∈(0, 1), recall its molliﬁcation at level √ t from (7). Then it is plain that ht is C∞with bounded derivatives of all orders and the solution to (10) with h = ht is given by ft(x) := −1 2 Z 1 t 1 1 −s(E[h(√sNΣ + √ 1 −sx)] −E[h(NΣ)])ds, see [29, p.12]. Finally, recall from e.g. [29, Lemma 2.2] that, for any t ∈(0, 1), dc(F, NΣ) ≤4 3 sup h∈Im \|Eht(F) −Eht(NΣ)\| + 20m √ 2 √ t 1 −t. Step 2 (integration by parts). An integration by parts by (5) and (4) (see [17, Chapter 4] for more details), together with Cauchy-Schwarz’s inequality, implies, Step 2 (integration by parts). An integration by parts by (5) and (4) (see [17, Chapter 4] for more details), together with Cauchy-Schwarz’s inequality, implies, \|Eht(F) −Eht(NΣ)\| = E d X i,j=1 Σ(i, j)∂2 ijft(F) −E d X k=1 Fk∂kft(F) (18) = E d X i,j=1 (Σ(i, j) −⟨DFi, uj⟩)∂2 ijf(F) ≤ v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2] v u u u t d X i,j=1 E[∂2 ijf(F)2]. (18) The following remarkable estimate is due to M. Schulte and J. Yukich. The following remarkable estimate is due to M. Schulte and J. Yukich. The following remarkable estimate is due to M. Schulte and J. Yukich. Lemma 2.1 (Proposition 2.3 in [29]). Let Y be an Rm-valued random vector and Σ be an invertible m × m covariance matrix. Then, Lemma 2.1 (Proposition 2.3 in [29]). Let Y be an Rm-valued random vector and Σ be an invertible m × m covariance matrix. Then, sup h∈Im E m X i,j=1 \|∂2 ijft(Y)\|2 ≤ Σ−1 2 op m2(log t)2dc(Y, NΣ) + 530m17/6 . where the left-hand side depends on h through the function ft solving Stein’s equation with test function ht given by (7). Remark 2.2. Lemma 2.1 improves upon the uniform bound (see [6] or [29]) \|∂2 ijft(x)\| ≤C(m, Σ) ∥h∥∞\| log t\|, when some a priori estimate on dc(Y, NΣ) is available. As consequence, when some a priori estimate on dc(Y, NΣ) is available. quence, \|Eht(F) −Eht(NΣ)\| ≤ Σ−1 op m\| log t\|dc(F, NΣ)1/2 + 24m17/12 v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2]. 2.1 Proof of Theorem 1.2 κ ≤4 3 Σ−1 op 2m 58( Σ−1 1/2 op + 1)m17/24 + 24m17/12 γ + 20 ≤ 4 3 × 140 × 2 Σ−1 3/2 op + 1 m41/24 + 20 √ 2m ! γ 3/2 / ≤402 Σ−1 3/2 op + 1 m41/24γ. The proof is complete. The proof is complete. 2.1 Proof of Theorem 1.2 ≤ Σ−1 op m\| log t\|dc(F, NΣ)1/2 + 24m17/12 v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2]. 9 9 Letting Letting κ = dc(F, NΣ), γ = v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2], κ = dc(F, NΣ), γ = v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2], γ = v u u u t d X i,j=1 E[(Σ(i, j) −⟨DFi, uj⟩)2], we have thus established we have thus established κ ≤4 3 Σ−1 op (m\| log t\|√κ + 24m17/12)γ + 20m √ 2 √ t 1 −t. (19) κ ≤4 3 Σ−1 op (m\| log t\|√κ + 24m17/12)γ + 20m √ 2 √ t 1 −t. (19) (19) Step 3 (exploiting the recursive inequality). If γ ≥1/e, then the bound we intend to prove holds trivially (observe that dc(F, NΣ) ≤1 by deﬁnition). Without loss of generality we can and will therefore assume that γ ≤1/e. Let t = γ2. Using the fact that κ ≤1 for the κ on the right-hand side of the (19), one has Step 3 (exploiting the recursive inequality). If γ ≥1/e, then the bound we intend to prove holds trivially (observe that dc(F, NΣ) ≤1 by deﬁnition). Without loss of generality we can and will therefore assume that γ ≤1/e. Let t = γ2. Using the fact that κ ≤1 for the κ on the right-hand side of the (19), one has κ ≤4 3 Σ−1 op (2m\| log γ\| + 24m17/12)γ + 20 √ 2mγ. Therefore, \| log γ\|√κ ≤ Σ−1 1/2 op r8m 3 γ1/2\| log γ\|3/2 + Σ−1 op 32m17/12 + 20 √ 2m 1/2 γ1/2\| log γ\|. Since supx∈(0,1/e] x1/2\| log x\|3/2 ≤4, one has \| log γ\|√κ ≤8 √ 6 3 Σ−1 1/2 op m1/2 + 16 √ 2 Σ−1 1/2 op m17/24 + 8 √ 10m1/2 ≤58 Σ−1 1/2 op + 1 m17/24. Hence, putting the estimate back into (19) with t = γ2 gives κ ≤4 3 Σ−1 op 2m 58( Σ−1 1/2 op + 1)m17/24 + 24m17/12 γ + 20 √ 2mγ ≤ 4 3 × 140 × 2 Σ−1 3/2 op + 1 m41/24 + 20 √ 2m ! γ ≤402 Σ−1 3/2 op + 1 m41/24γ. 2.3 Proof of Corollary 1.4 We follow closely the arguments of [21] and assume without loss of generality that T = 1. First, one can embed the Gaussian sequence in the statement in an isonormal Gaussian process {X(h) : h ∈H}, in such a way that {Gk : k ∈Z} d= {X(ek) : k ∈Z} , {Gk : k ∈Z} d= {X(ek) : k ∈Z} , for some appropriate family {ek} ⊂H verifying ⟨ej, ek⟩H = ρ(k −j) for all j, k. For ϕ = P ℓ≥d aℓHℓ∈L2(R, γ), we deﬁne the shift mapping ϕ1 := P ℓ≥1 aℓHℓ−1 and set un,i := 1 √n ⌊nti⌋ X m=⌊nti−1⌋+1 ϕ1(Gm)em, i ∈[m]. Then, standard computations using (6) lead to Then, standard computations using (6) lead to δ(un,i) = Fn,i. (20) δ(un,i) = Fn,i. (20) Applying Theorem 1.2 and the triangle inequality implies that Applying Theorem 1.2 and the triangle inequality implies that dc(F, NΣ) ≤C m X i,j=1 \|Σ(i, j) −E[FiFj]\| + C v u u t m X i,j=1 Var(⟨DFn,i, un,j⟩) =: I1 + I2. Note that, by the chain rule and the relation D(Gk) = ek, Note that, by the chain rule and the relation D(Gk) = ek, Note that, by the chain rule and the relation D(Gk) = ek, ⟨DFn,i, un,j⟩H = 1 n X k∼ti X ℓ∼tj ϕ′(Gk)ϕ1(Gℓ)ρ(k −ℓ), where k ∼ti means that the sum is taken over k ∈{⌊nti−1⌋+ 1, ..., ⌊nti⌋}, and similarly for the symbol ℓ∼tj. Hence, where k ∼ti means that the sum is taken over k ∈{⌊nti−1⌋+ 1, ..., ⌊nti⌋}, and similarly for the symbol ℓ∼tj. Hence, Var(⟨DFn,i, un,j⟩H) = 1 n2 X k∼ti X ℓ∼tj X k′∼ti X ℓ′∼tj Cov(ϕ′(Xk)ϕ1(Xℓ), ϕ′(Xk′)ϕ1(Xℓ′))ρ(k −ℓ)ρ(k′ −ℓ′) ≤ 1 n2 n X k,k′,ℓ,ℓ′=1 Cov(ϕ′(Xk)ϕ1(Xℓ), ϕ′(Xk′)ϕ1(Xℓ′))ρ(k −ℓ)ρ(k′ −ℓ′) . (21) ≤ 1 n2 n X k,k′,ℓ,ℓ′=1 Cov(ϕ′(Xk)ϕ1(Xℓ), ϕ′(Xk′)ϕ1(Xℓ′))ρ(k −ℓ)ρ(k′ −ℓ′) . (21) (21) The variance is bounded because of the assumption that ϕ ∈D1,4. Once (21) is in place, one can apply Gebelein’s inequality as in [21]. In particular, one infers that (see [21, Proposition 3.4]) The variance is bounded because of the assumption that ϕ ∈D1,4. Once (21) is in place, one can apply Gebelein’s inequality as in [21]. In particular, one infers that (see [21, Proposition 3.4]) dc(Fn, NΣ) ≤C v u u t 1 n2 n−1 X i,j,k,ℓ=0 ρ(j −k)ρ(i −j)ρ(k −ℓ) . 2.2 Proof of Corollary 1.3 We will obtain the desired conclusion as a direct application of Theorem 1.2 with ui = −DL−1Fi, see (6). Indeed, recall that by Step 2 of [22, Proof of Theorem 4.3], for any i, j ∈[m], E[(E[FiFj] −⟨DFi, −DL−1Fj⟩)2] ≤Cov(F 2 i , F 2 j ) −2E[FiFj]2. and, Step 3 of [22, Proof of Theorem 4.3] shows that E[(E[FiFj] −⟨DFi, −DL−1Fj⟩)2] ≤Cov(F 2 i , F 2 j ) −2E[FiFj]2. E[(E[FiFj] −⟨DFi, −DL−1Fj⟩)2] ≤Cov(F 2 i , F 2 j ) −2E[FiFj]2. On the other hand, Step 3 of [22, Proof of Theorem 4.3] shows that m X Cov(F 2 i , F 2 j ) −2E[FiFj]2 = E ∥F∥4 −E ∥NΣ∥4 . On the other hand, Step 3 of [22, Proof of Theorem 4.3] shows that m X i,j=1 Cov(F 2 i , F 2 j ) −2E[FiFj]2 = E ∥F∥4 −E ∥NΣ∥4 . Plugging these estimates into Theorem 1.2 gives the result. Plugging these estimates into Theorem 1.2 gives the result. 10 2.3 Proof of Corollary 1.4 If, in addition, ϕ is 2-sparse, then If, in addition, ϕ is 2-sparse, then dc(Fn, NΣ) ≤C v u u t 1 n2 n−1 X i,j,k,ℓ=0 ρ(j −k)2ρ(i −j)ρ(k −ℓ) . 11 11 Items i)-ii) now follow from these inequalities, as shown in [21]; we include a proof for complete- ness. Applying twice Young’s inequality for convolutions, one has, with ρn(k) = \|ρ(k)\|1\|k\|<n, Items i)-ii) now follow from these inequalities, as shown in [21]; we include a proof for complete- ness. Applying twice Young’s inequality for convolutions, one has, with ρn(k) = \|ρ(k)\|1\|k\|<n, n−1 X i,j,k,ℓ=0 ρ(j −k)ρ(i −j)ρ(k −ℓ) ≤ n−1 X i,ℓ=0 ρn ∗ρn ∗ρn (i −ℓ) ≤ n ∥ρn ∗ρn ∗ρn∥ℓ1(Z) ≤n ∥ρn∥3 ℓ1(Z) , n−1 X i,j,k,ℓ=0 ρ(j −k)ρ(i −j)ρ(k −ℓ) ≤ n−1 X i,ℓ=0 ρn ∗ρn ∗ρn (i −ℓ) ≤ n ∥ρn ∗ρn ∗ρn∥ℓ1(Z) ≤n ∥ρn∥3 ℓ1(Z) , yielding Item i). Rewrite the sum of products as a sum of the product of convolutions by introducing the function 1n(k) := 1\|k\|<n. We have yielding Item i). Rewrite the sum of products as a sum of the product of convolutions by introducing the function 1n(k) := 1\|k\|<n. We have n−1 X i,j,k,ℓ=0 \|ρ(j −k)2ρ(i −j)ρ(k −ℓ)\| = n−1 X i,j,k,ℓ=0 \|ρ(j −k)2ρ(i −j)ρ(k −ℓ)1n(ℓ−i)\| ≤ n−1 X j,ℓ=0 (ρn ∗1n)(ℓ−j)(ρn ∗ρ2 n)(ℓ−j) ≤n⟨ρn ∗1n, ρn ∗ρ2 n⟩ℓ2(Z). ≤ n 1 X j,ℓ=0 (ρn ∗1n)(ℓ−j)(ρn ∗ρ2 n)(ℓ−j) ≤n⟨ρn ∗1n, ρn ∗ρ2 n⟩ℓ2(Z). For b ∈[1, 2], we have For b ∈[1, 2], we have ⟨ρn ∗1n, ρn ∗ρ2 n⟩ℓ2(Z) ≤∥ρn ∗1n∥ ℓ b b−1 (Z) ρn ∗ρ2 n ℓb(Z) (22) ≤∥ρn∥ℓb(Z) ∥1n∥ ℓ b 2b−2 (Z) ∥ρn∥ℓb(Z) ρ2 n ℓ1(Z) ≤(2n) 2b−2 b ρ2 n ℓ1(Z) ∥ρn∥2 ℓb(Z) , (22) yielding Item ii). Now we move to the proof of Item iii). Notice that taking b = 2 for the right-hand side of (22), together with an application of Young’s inequality, yields that yielding Item ii). Now we move to the proof of Item iii). Notice that taking b = 2 for the right-hand side of (22), together with an application of Young’s inequality, yields that ⟨ρn ∗1n, ρn ∗ρ2 n⟩ℓ2(Z) ≤∥ρn ∗1n∥ℓ2(Z) ∥ρn∥3 ℓ2(Z) . Thus, 1 n2 n−1 X i,j,k,ℓ=0 ρ(j −k)2ρ(i −j)ρ(k −ℓ) ≤1 n ∥ρn ∗1n∥ℓ2(Z) ∥ρn∥3 ℓ2(Z) . To proceed, we handle the convolution involving 1n a bit diﬀerently. Set To proceed, we handle the convolution involving 1n a bit diﬀerently. A Proof and discussion of relation (3) Inequality (3) is a direct consequence of the following statement, whose proof exploits a strategy already adopted in [2, Proof of Theorem 3.1]. Proposition A.1. Fix m ≥1, and let NΣ denote a m-dimensional centered Gaussian vector with invertible covariance matrix Σ. Then, for any m-dimensional random vector F one has that hat dc(F, NΣ) ≤2 √ 2 Γ(Σ)1/2 dW (F, NΣ)1/2, (23) h Γ(Σ) i th i i t i t t d ﬁ d b dc(F, NΣ) ≤2 √ 2 Γ(Σ)1/2 dW (F, NΣ)1/2, (23) metric constant deﬁned by dc(F, NΣ) ≤2 √ 2 Γ(Σ)1/2 dW (F, NΣ)1/2, (23) (23) where Γ(Σ) is the isoperimetric constant deﬁned by Γ(Σ) := sup Q,ϵ>0 P(NΣ ∈Qϵ) −P(NΣ ∈Q) ϵ , where Q ranges over all Borel measurable convex subsets of Rm, and Qϵ indicates the set of all elements of Rm whose Euclidean distance from Q does not exceed ϵ. Remark A.2. In [14] it is proved that, for some absolute constants 0 < c < C < ∞, c q ∥Σ∥H.S. ≤Γ(Σ) ≤C q ∥Σ∥H.S., where ∥· ∥H.S. stands as above for the Hilbert-Schmidt norm. When Σ = Im (identity matrix), one has also the well-known estimate Γ(Im) ≤4m1/4 (see [3]), as well as Nazarov’s upper and lower bounds ( ) ( ) where ∥· ∥H.S. stands as above for the Hilbert-Schmidt norm. When Σ = Im (identity matrix), one has also the well-known estimate Γ(Im) ≤4m1/4 (see [3]), as well as Nazarov’s upper and lower bounds e−5/4 ≤lim inf m Γ(Im) m1/4 ≤lim sup m Γ(Im) m1/4 ≤(2π)−1/4 < 0.64, see [14, p. 170]. In [28, Theorem 1.2], it is proved that Nazarov’s upper bound can be reduced from 0.64 to 0.59; see also [4] for related computations in the framework of the multivariate CLT. see [14, p. 170]. In [28, Theorem 1.2], it is proved that Nazarov’s upper bound can be reduced from 0.64 to 0.59; see also [4] for related computations in the framework of the multivariate CLT. Proof of Proposition A.1. We can assume that F and NΣ are deﬁned on a common probability space, and that E∥F−NΣ∥Rm = dW (F, NΣ). Fix a convex set Q, as well as ϵ > 0. 2.3 Proof of Corollary 1.4 Set eρn(k) = ρ(k)1N≤\|k\|<n, bρn(k) = ρ(k)1\|k\|≤N eρn(k) = ρ(k)1N≤\|k\|<n, bρn(k) = ρ(k)1\|k\|≤N so that ρn = eρn + bρn. One has 1 n ∥ρn ∗1n∥ℓ2(Z) ≤1 n ∥eρn∥ℓ2(Z) ∥1n∥ℓ1(Z) + 1 n ∥bρn∥ℓ1(Z) ∥1n∥ℓ2(Z) ≤ X N≤\|k\|<n ρ(k)21/2 + (2N + 1)n−1/2, 1 n ∥ρn ∗1n∥ℓ2(Z) ≤1 n ∥eρn∥ℓ2(Z) ∥1n∥ℓ1(Z) + 1 n ∥bρn∥ℓ1(Z) ∥1n∥ℓ2(Z) ≤ X N≤\|k\|<n ρ(k)21/2 + (2N + 1)n−1/2, from which Item iii) follows. The proof is complete. 12 A Proof and discussion of relation (3) The presence of the factor (log m)1/4 is consistent with the fact that, for the standard Gaussian measure on Rm, the isoperimetric constant associated with all hyper-rectangles of Rm is bounded from above by √log m, see [3, 14]. An estimate analogous to (24) is established by diﬀerent methods in [12, Corollary 3.1]. [13] S.V. Nagaev (1976). An estimate of the remainder term in the multidimensional central limit theorem. In: Proceedings of the Third Japan–USSR Symposium on Probability Theory (Tashkent, 1975). Lecture Notes in Math. 550, 419–438. Berlin: Springer. A Proof and discussion of relation (3) We have that P[F ∈Q] −P[NΣ ∈Q] ≤ P[F ∈Q, ∥F −NΣ∥Rm ≤ϵ] −P[NΣ ∈Q] + ϵ−1E[∥F −NΣ∥Rm] ≤ P[NΣ ∈Qϵ] −P[NΣ ∈Q] + ϵ−1dW (F, NΣ) ≤ Γ(Σ)ϵ + ϵ−1dW (F, NΣ). On the other hand, deﬁning Q−ϵ as the set of those y ∈Q such that the closed ball with radius ϵ centered at y is contained in Q, On the other hand, deﬁning Q−ϵ as the set of those y ∈Q such that the closed ball with radius ϵ centered at y is contained in Q, P[NΣ ∈Q] −P[F ∈Q] ≤ P[NΣ ∈Q, ∥F −NΣ∥Rm ≤ϵ] −P[F ∈Q−ϵ] + ϵ−1E[∥F −NΣ∥] ≤ Γ(Σ)2ϵ + ϵ−1dW (F, NΣ), where we have used the inequality P[NΣ ∈Q, ∥F −NΣ∥Rm ≤ϵ] −P[F ∈Q−ϵ] ≤P[NΣ ∈Q] −P[NΣ ∈Q−2ϵ]. The conclusion follows from a standard optimisation in ϵ. 13 Remark A.3. Fix m ≥1, and let Rm be the collection of all hyper-rectangles of the type R = (−∞, t1] × · · · × (−∞, tm]. In [2, Theorem 3.1] it is proved that, if N is a m-dimensional centered Gaussian vector with identity covariance matrix and F is any m-dimensional random vector, then Remark A.3. Fix m ≥1, and let Rm be the collection of all hyper-rectangles of the type R = (−∞, t1] × · · · × (−∞, tm]. In [2, Theorem 3.1] it is proved that, if N is a m-dimensional centered Gaussian vector with identity covariance matrix and F is any m-dimensional random vector, then sup R∈Rm P[F ∈R] −P[N ∈R] ≤3(log m)1/4dW (F, N)1/2. (24) sup R∈Rm P[F ∈R] −P[N ∈R] ≤3(log m)1/4dW (F, N)1/2. (24) The left-hand side of the previous inequality is usually referred to as the Kolmogorov distance between the distributions of F and N. The presence of the factor (log m)1/4 is consistent with the fact that, for the standard Gaussian measure on Rm, the isoperimetric constant associated with all hyper-rectangles of Rm is bounded from above by √log m, see [3, 14]. An estimate analogous to (24) is established by diﬀerent methods in [12, Corollary 3.1]. The left-hand side of the previous inequality is usually referred to as the Kolmogorov distance between the distributions of F and N. 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Zheng (2010): Multi-dimensional Gaussian ﬂuctuations on the Poisson space. Electron. J. Probab. 15, 1487–1527. [28] M. Raic (2019): A multivariate Berry–Esseen theorem with explicit constants. Bernoulli 25(4A), 2824–2853. [29] M. Schulte and J.E. Yukich (2019): Multivariate second order Poincaré inequalities for Poisson functionals. Electron. J. Probab. 24, no. 130, pp. 1–42. [30] C. Villani (2009). Optimal transport. Old and new. Grundlehren der mathematischen Wis- senschaften 338. Springer: Berlin. [31] N. Zygouras (2019). Discrete stochastic analysis. Lecture notes available on the webpage: https://warwick.ac.uk/fac/sci/statistics/staff/academic-research/zygouras/ 15	17,341
W2801636571.txt_3	German-Science-Pile	Open Science	Various open science	null	None	None	German	Spoken	7,730	19,178	Auch nach 56) sei die in neue sondern der im Themen der pro- im ideologischen auf die Mas- Partei bei der Ver- a lle r die Aufgaben. Organi- Vereinigungen pro- VOAPP (Vsesojuznoe P is a te le j) , die VAPP be- von der P roletarskich P is a t e le j) , im allgemeinen c h a r a k te r is ie r t Zunächst diese land, Kampf was auch Unterscheidung den Grundlinien s o ll auf Im Übrigen beschränkt tags le h ie r RAPP“ w ird. Associacija der Einfluß der geschaffen, besseren in VOAPP» in deren Averbach b lie b. 1928, der zur (Russkaja wie g e s te llte n S c h r i f t s t e l 1er nannte Elemente» führenden Rolle Instrument le ta ris c h e r kennzeich- “ Abweichungen einzelner im formalen Partei des tru g. schonungsloser ihrem vorherrsehenden S c h r i f t s t e ll e r w irklichung Erringung und Averbachs Entwicklung “ neubürgerliche" d ie - p ro letarischen Resolution beeinflussen: “ Schwankungen“ bedeutete; als diese mit Schlußresolution, Handschrift es, sich Im A p ril/M a i der eine sind “ poputbiki" der id e n tifiz ie r te. verabschiedete gegen von VAPP, Allunlons-Kongreß entscheidend der Bereich der bedeutendem Maße die die Sowjetunion FUhrer P a rte i s ta tt. zentrale nen der und mit wird» sich damit t io n zu fördern. die eingeschatzt Z e it "Herrschaft und bewertet die nach dem W illen des XV. auf das Sowjet- Sowjet 1i t e r a t u r auch dort dem Stichwort die L ite r a tu r Rahmen wird n ic h t auch eine zugewiesen, Werken hineinzuwirken und wie Bewertungen eingegangen werden, Aufgabe der in te rn a tio n a le n d a rg e s te llt, in die P a rte i- V o r r e it e r r o l- nämlich mit ihren k a pDiether ita lis c h e Welt Götztis - 9783954791927 Sache der proletarischen WeltrevoluDownloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 28 aus den 30er und 40er zu bezeichnet nennen, Entwicklung tio n of nach kommt zur (1929-1932), easy W riters as a immediate p u t& ik i, ments Anm. of the Entwicklung whether W as RAPP"60 maßen: in W riters gram that the war w ith in ra tu re. depiction its and outlaw ed."61 a ll plan e in g e re ih t: sie in 57) G. vonic 58) 59) Struve, G. P. Reavey, 60) E. 61) Simmons, (1880-1940), 62) V. die only Soviet Soviet 1942, Setschkareff, o u tlin e N.Y. Geschichte by a c tiv ity on Russian w ri- reign folgender- c r itic Averbakh pro- and of the class Soviet lite - execute t h e ir of to found in themselves der Wert, gesteckten wenn in - b e itra g e n ." 62 Congress, Nr. der den FUnfJahres- haben nur Z ie le of uncompromising Erlangung 39, in : 1935, New Haven S. The Sia641. 1947, Culture. S. Part 17. II: 117. S. modern S. "... Pro- L ite r a tu r 1949, of 1943, lit e r a r is c h e auch Setchkareff, Russian S. achieve- of Russian L ite ra tu re , Cambridge/Mass. An the lin e sich Today, of on the problem L ite ra ry L ite ra tu re Outlines [po- S te lle Plan hew to w irts c h a ft 1ichen Ithaca w ith Erzeugnisse The Pan-Soviet Travellers d ic ta to r ia l down the äußert Weise zur Philadelphia Studies, to that Russian Association European Review X I I I , Simmons, Slavic E. Jener M iljukov, L ite r a tu re , Sinne lite r a r is c h e oder "the Five-Year was the * 1929 wird oder and East the was die campaign was promulgated Im selben nenpoli tisehen of la id it among the an anderer the who refused fe s ts te llt: der and framework An extensive program, dazu (RAPP) themselves in s is te d sieht free P ro le ta ria n and by vigorous er of of 1928 gekennzeichnet, als Period comparatively , r e p o r t’ communists, control P la n ." 57 Year Plan Fellow elements einmal interested le ta r ia n to nach e rlä u te r t seized Five and Auch Miljukov p ro le ta ria n bezeichnet, head, others, pressure Simmons ‫״‬A few the or dem Zeitraum of p r o le t a r ia n isa- e sta b lish in g w rite rs , V e r f.] the in of the t e r s . “ 59 succeeded lit e r a r is c h e Five-Year the fie ld governmental part the lit e r a r y increased die "... the with RAPP beim Namen The Russian Association P la n ."5• in "... the duty d. der with sharp break now in Z eit compliance period. Ohne die 1935 Struve eine F eststellung: (R.A.P.P.) d ic ta to r s the in made Nep stammen. b e re its 1928 a ls lit e r a t u r e Reavey and Jahren in : Handbook of Russian lit e r a t u r e. 552. 49. ..., S. 131. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 29 Für westliche 1928 L ite ra tu rw is s e n s c h a ftle r das Ende der der r e la t iv Sowjet 1it e r a t u r. gemacht» und die fre ie n OafUr RAPP a ls bedeutet lite ra ris c h e n wird die Z e it nach Entwicklung P artei deren w illfä h r ig e s die in verantwort 1ich Instrument ange- sehen. Betrachtet bung der der man 30er und RAPP nicht RAPP im sehen Standpunkt Jahre Dies gen 40er aus, bedeutet, dabei die wird zur man in ih re Vorwürfe, die 30er 40er und die indes Jahre te o r ii der Kunstwerkes 1951 sowjetische a u ftr itt? vermerkt Die gii svoej bleibe bei der RAPP."41 Damit v Uber vorgeworfen, sich so: lite r a - im Zusammen- V o rs te llungen auch a ls das We- sowie die Wesen eines h in te r dem B e g r iff " v u l’ garnyj Beziehung zu setzen 63) M. Rozental Moskva otricaet S. Vorwurf, aus s o c io lo dem Jahre podlinnye ... І sociolo- "V ulgärsoziolo- und wehre sich zu den unveränderlichen 0 6. sovetskaja der g e s e lls c h a ftlic h e r Oberfläche (iz d .)» 1936, der Analyse in Bol’ &aja s o c io lo g i! sich special'noj cel'Ju bor’ bu Zusammenhang steht der 402. lesen istorifceakogo aaterializaa."4* an S. P a rte i der oblKestvennogo razvitija Entwicklungen 64) dem die Literaturw issenschaft tipiUny• dija burXuaznoj tavja&Kej protiv s t a t 'i, Würdigung, Weise das o b je ktive der westlichen Die v u l'g a rn o j sociologi■ javljaetsja gismus" f e s t g e s t e llt negative mit lie - hierzu: Vul’ garnyj in oben "Große Sowjet-Enzyklopädie" zakonoaernosti Damit A lle rd in g s ausüben konnte. W as v e rb irg t so cio lo ģ ija "» der L ite ra tu rw is s e n - RAPP bei sah, und des Kunstwerkes m arxistischer wo die sondern aus RAPP erhebt, v period aufzudecken. " v u l ‫״‬garnaja der Bereich die Jahre, RAPP 1932 geführt wie eine RAPP unmarxistische in der anders: Instrument v lija n ie war, b e u r t e ilt. T ä tig k e it gegen der T ä tig k e it von jenem 1І te r a tu r p o li t i - deshalb k r it ik e L ite r a tu r U nfähigkeit, gizm" bylo i hang wurden der sen die daß die sowjetische im lite r a r is c h e n " Osobenno s i l 'n o tu rn o j führend Auflösung v ö llig das w i l l i g e Herrschaft Sicht RAPP negativ fand ih r aus der daß auch die L ite ra tu rw isse n sch a ftle rn w eil auf, konkret, der Akzente konnte, L ite ra tu rg e s c h ic h ts s c h re i- so f ä l l t Bereich danach» das Wirken der werden Jahre, 1ite ra ris c h e n der schaft sowjetische b e u r t e ilt Sicht hat. die sovetskoj lit e r a t u r e. Fakten und dagegen, sie GesetzmäßigK r itib e s k ie Diether Götz - 9783954791927 Downloaded from tom PubFactory 01/10/2019 ènciklopediJa, 9, at M oskva03:46:08AM 1951, via free access 00050394 30 keiten der h is to ris c h -g e s e lls c h a ft 1ichen Bewertung der RAPP die a ls Anwalt sich stand, wird in jener verbunden der und s te llt sredi zu RAPP und fe s t: zu Vollmachten So übte zu die n ic h ts anderes ü brig , oder A lle rd in g s finden des L it e r a t u r in A. 19. 66) G. 67) W. 21958, die Struve, in sich der in den 50er der RAPP " . .. d ik ta to ris c h e Sinne der T ä tig k e it habe eine sovetskoj ..., Russische S. L ite r a tu r sich Letten- RAPP gegen andere Feststellung b lie b Betätigung auch schon d iffe r e n - der RAPP. der Zwar weist RAPP-Leute und auch auf s te lle n. der Tat- habe, die Andererseits Versteifung lite r a tu r y , in - die L ite r a tu r th e o r ie mit zu v e r u r te ile n . " 6 7 id e n tifiz ie r t sie Geschichte der a lle zu Wirken zu sich S c h r if t s t e lle r n Jahren RAPP, le t in äußert selbst Partei T rid c a t' nach der lite r a r is c h e n [h a tte ] Jahren Jahren vorge- L i t e r a t u r lebens gemeinsam 50er L ite ra tu rw isse n ‫־‬ wenn s ie dem Z ie l gegen die den 40er der der den abschließenden Werke die haben. RAPP in Dienst Lettenbauer, S. hätten worin Attacken P o lit ik , hatten, RAPP mit "wenig Egolin, die darauf, geführt die das d ik ta to ris c h e den er daß sich rabotu westlichen schon früheren sich daß sich bescheinigt w ic k e lt, der und zur als sowjetischen hin, so v o s p it a t e lnuju RAPP angegriffenen ihre auf Egolin, sich pisa- zunehmende Polemik Betrachtungsweisen Simmons sie bast scharfen Im ähnlichen d a r s te llt enthalten der versammelt Hinweis um die Struve ric h te n von der S. nach w o llt e n . “ 44 er habe, ver- ideologischen znabi t e l ‫״‬nuju Auflösung die ‫״‬Den 65) ih r e r L in ie , der kommt: sache zu den T e il S c h r ifts te lle r nerhalb die ein wenn die des Volkes" einen aus und bestimmte weiterleben auch g ib t b le ib t S c h r ifts te lle r gegen to r m o z ili Diskussion war. Organisation, L ite r a tu r p o litik 1948 k o n s ta tie r t man die zeichnet der verstoßen " I z o lir u ja Satz le tz tlic h auf eben diese Diese 1it e r a t o r o v . 6 ‫ ״‬ѣ le tz te s c h a ftle r z ie r te r e sie samym sind, so bauer, daß Noch suchen V e rfo lg t w e iter, in P a rte i tem massy Gerade dieser Gründe P artei RAPP "Feinde w eiter rappovcy p a r tii der fU h lt. FUhrung der te le j, erstaunen; beschuldigt, Glaubenssätze eng muß Entwicklung. entder Moskva 1948, 255. L ite ra tu rg e s c h ic h te , Wiesbaden 270. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 31 P artei auf Vorzüge d. den Vorsatz, des V erf..1 sohaft der Planes zu RAPP eine ‫״‬Wegreißen der seine dieser Die re ... Masken* des a ls b is hin scharfen «auf der ih r e r sowie aut K r itik lin 1930 in dert h a tte : sich der der Menschen* und und dem und so- Tugenden e n t h ü llt e . * 0* Simmons übrigens pisatelej RAPP, der RAPP gegenüber wobei Nichtbefolgung zur In auch in ... Schwerpunkt nun Sehri f t te 1- lie g t, Sta- g e fo r- partijnych ne ob-javljali vsjaëeski гаХіяаІІ v sovetskoj saaokritiku die Bereichen Ł vsech pisatelej, ïe Rappovey i bei ukazanijaai prinadleüavlich к ich gruppirovke, kritiku Jah- Parteianweisungen a lle n prenebregali 50er ebenfalls anderen der Oni proti vopostavljail bespartijny■ der S e lb s t k r it ik an G o r 'k ij R A P P a der KPdSU b le ib t der ‫־‬Rukovoditeli ... der an einem B rie f ѵгадаві Anm. FUhrer- Lebens widmete RAPP s ie h t Unfähigkeit partii die die re a lis tis c h e n lebendigen seine fü r Fünfjahresplan, v e r te id ig te sowjetischen auch angeblichen ih re r des P a rte ita g Unduldsamkeit le rn , 1. Propaganda Literaturgeschichtsschreibung zun! XX. der die zur Auflösung. 69 deren sowjetische der Weiterhin Darstellung Laster fü r is t L it e r a t u r , "Vermessenheit“ den Grund L it e r a t u r [gemeint benützen psychologischen wohl die literature. ' ‫ י‬ 68) E. Simmons, S tu ttg a rt S. Through the society, New York 69) Borland H. glass Diskussion w rite lis m . * H. of Soviet macht F ir s t Five-Year Dieser Gedanke der Methode des und dabei ‫״‬ die theoretische und es wird byl 70) Pospelov, A. dlja der T e orija Dementev Russian Tatsache the d ia le c t ic a l w r ite r materia- and Practice 1950, 1940 bei Pospelov, L it e r a t u r daß dieser auf sei RAPP sehr bald during 143. wenn er Grundlage schaffen B e g riff Deshalb S. der stammen und sich diese w o ll- aus dem abn ic h t eignen lite r a tu r - k r it is ie r t worden, g e fo lg e rt: oboznafcenija bolee ..., ( i. 1951, die New York osobennostej ■ySlenija v social istiKeskoj vydvinut Leningrad/Moskva Sirnmons, of that Materialismus" Bereich L ite r a tu r. auf Theory schon daß sie würden, ’ socialistiüeskij G. E. 1i t e r a t u r t h e o r e t isehen method of d iale ktisch e n Auffassung ” ... the L ite r a r y erscheint von Pospelov skogo Sowjet 1i t e r a tu r , von: views demanded 1929-1932, v o rh ä lt, verkennen fü r to Plan der RAPP leaders Soviet s t r a k t - p h ilosophischen würde L ite r a tu r e : der according RAPP-Leuten der Übersetzung h in s ic h t lic h M RAPPs Borland, den im Spiegel Deutsche 1953. daß should ten Mensch 18. innerhalb aufmerksam, the Der 1956, specifi&eskij terain rea liz■ '.” S. 325. d r.), S. tvorfce- literature 224. Russkaja sovetskaja lit e r a t u r a , Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access In derselben vyd vin u li p o lit ib e s k i raskalyvajo Wendet durch man sich westliche Äußerungen, RAPP heraussteilen of d ic ta to r in the T ä tig k e it an. Dagegen die wie realm der und auf in den der W irk lic h k e it der K o n flik t verlangte, RAPP-Theoreti k e r , fand, die Forderung kann wurde die als d. sie Mehrzahl der die s o z ia lis tis c h e “ lebenswahr" Erkenntnis in te r p r e ti e r t durch, Oberk 72) H. Muchnic, From Gorky 73) A. Préchac, La L it t é r a t u r e lin s K.-D. is to r ii ..., Seemann, Tod in Osteuropas zu S. Die - wie wobei sich den RAPP- fUr die und seien die die zu Ende Wirk- von daher Schilderung “ revolutionären sie is t: "Deren [der künstlerische Methode, die in wie Menschen" W irk lic h k e it ihren Ausdruck dem Sinne bezeichnen, zu schildern der sei ... kommunistischen RAPP f e s t h i e l t. R ealist müsse daß es is t K r it ik e r Immer wieder imstande sein, s c h ild e r n ." 74 Zunehmend setzt T ä tig k e it Ausdruck der der RAPP nicht S e lb s th e rrlic h k e it 297. to Pasternak, S icht, N. 13 New York Soviétique, sowjetische h is to ris c h e r (Neue Reihe), Jahren die daß die werden kann a ls 71) 74) der c u l t u r e l " 71 die n ich t is t, sehen RAPP im Rahmen der hin, hätten der Asso- Wertungen im Sinne der V e rf.] an den Vorstellungen betont, Jahre immer an absolute Sowjetunion geraten, r e a lis t is c h die der sich All-Russian den 60er Schilderung, s o ll, so wie daß W irk lic h k e it sich sein in und 70er Regiment "terrorism e eingesetzt P a rte i eine finden solche RAPP d i f fe re n z ie r- became . .. in nach dem “ lebendigen W irk lic h k e it, zunächst als sie L it e r a t u r Anm. man beachtenswert, RAPP, 30er wird, der der sie der vrag’ , der den 60er the Bedeutung der L it e r a t u r mit in wie "and Diskussion Anfang in Zwar b e re its die T ä tig k e it Zunahme der l i t e r a t u r e " 72 ; a tte s tie r t in v o r s t e llt , W rite rs, i l i ... p i s a t e l e j . " 71 das d ik ta to ris c h e diese: Seemann zu 1ichkeitstreue eine "Rappovcy ‫״‬sojuznik Bewertung der RAPP le d ig lic h weist Theoretikern der etwa of lite ra tu rth e o re tis c h e n Romantik" der le d ig lic h schon Simmons - 20er sovetskich fe s t s t e lle n. P ro le ta ria n die der lózung so kann man generell c ia tio n F eststellung: Literaturw issenschaft i e r Betrachtungsweisen auch die vrednyj wieder noch in lie g t tem samym rja d y Jahren zu, ten Richtung о. О., 1961, S. 1977, S. L ite r a tu r p o li t i k in : Jahrbücher (1965), S. fü r s e it 23. 56. Sta- Geschichte 42. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 33 dieser F a ll Organisation, le tz te n d lic h m uß also und und Thomson RAPP tio n a l ‫״‬On classless tated c u ltu re , in fie d p o lit ic a l shows c le a rly P a r ty ." 7* th a t verwahrte sich in den rung nach zur were charged w ith when Soviet the ... the there das Fragen der K r it ik e r n einer reinen der fo r which sich advocated they of the scharf, auf S eit Einmischung the had d ie - standards auch auf by the den um Unabin lit e r a t u r die aber RAPP ja ‫״‬Averbach inkompetenten Funk- Theorie indem er ju s ti- Vorgänge um is t geraten, in RAPP a f f a i r die 1930 von a had indeed been RAPP-FUhrung K r itik fac- was an ominous note wird 1ite ra ris c h e n sehr Propagandakunst der s te llt accused campaign was engineered der der was entering lite r a r y hingewiesen. gegen die having The aftermath D eutlich Schußfeld Hierzu p o s itio n society l it e r a t u r e but dessen E xi- e d it o r ia l The RAPP leaders whole RAPP. Fragen in tete der s te h t. a Pravda ih r Charakters Zusammenhang mit th e ir bemühten Standpunkt zunehmend tionären 1932, auch " RAPP-Diktatur‫״‬ Phänomen, P artei Bemerkungen beziehen Liquidierung theoretischen ein Pravda assumed th a t Diese hängigkeit a ls abused stage; action Die aber in s tru m e n ta lis ie rte n der d i c t a t o r i a l, which KPdSU. unmittelbarem illite r a te. and A u fstie g , having ju s t s o c ia lis t in to le ra n t die of they was ca lle d way in 23 A p r il leadership in te re s ts ; von der werden, Interessenlage p ro le ta ria n the a n a ly s ie rt fe s t: daß ih r ih re s Verschwinden jew eiligen the abhingen h in s ic h t lic h gesehen stenz sondern ... [und] in d ir e k t A g it a t io n s lit e r a t u r in antwor- die eine Forde- Beziehung deutschen N a tio n a ls o z ia lis te n brach- t e / 7* Auf ganz andere der Sowjetunion die sowjetischen XX. P arteitag z ie h t. Einige fen. So kommt 75) B. 76) P. in den Jahren KPdSU Beispiele etwa S. b is s o lle n Trifonova zu The Premature 1917-1946, Hübner, 1ite ra ris c h e n 1928-1932 Veröffentlichungen der Thomson, and Society Sehweisen der 212f. London stößt in S. wenn man Jahre heran- davon verschaf- F eststellung: Revolution. 1972, 60er Eindruck folgender man, in den Jahren nach dem zum Ende der einen Entwicklung Russian L ite ra tu re 224. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access Pobeda socielistifceskogo realizza ne byla legkoi pobedoj. O na byla dostignuta v chode idej- noj i borby iskov naprjaïennych 20-ch godov ... loSit’ tvor£eskich po- Olja togo, ütoby pro- dorogu sociallfttl&eskogo realizna, bylo osvobodlt' sJa teorij, ... ot istolkovanija ot nado foralisti&eskich vul' garno-sociologlfceskogo klassovoj prirody iskusstva, ot Mechanistibeski-p ria itivnogo poni■an!ja svjazi iskusstva s H zn'ju.“ 7 7 Bei Timofeev/Dement ev heißt 'V es: sostojavSejsja besede pisatelej posle s likvidacii R A P P a rukovoditeljani partii 02e$tvennyj ■etod sovetskoj literatury byl chud opredelen как ■etod 30cialisti£esk0g0 realizza. Ū L FUr den genannten Zeitraum iskusstvennaja rye podbas die setzung h ie r f ü r "... ot sei s te llt es turwissenschaft gische es als dar, genauso gegemonija fa lls große monie" Rolle der der 77) Jahre T. Leistung H ilfe der Pospelov der der Ansatz Partei um folgenden Trifonova, L. skoj lit e r a t u r y. S. Timofeev/A. 79) V. Kovalev, S. Asadullaev, RAPP G. n a tü rlic h der überwunden werden "No teoretiKeskaJa wobei dann eben- diese In h a ltlic h is t die s o w je ti- der 50er und der russkoj sovet- zweiten H ä lfte ‫״‬Hege- Istorizm , Baku ( iz d . ) , 1929-1941. 1969, . .., S. t e o r ija S. p a r t i ja I s to r ija lioskva Sovetskoe 1960, S. 8. 77. i t ip o lo g ija s o c ia lis ti- 48. i p a r t ijn a ja o o l’ &uju pefcat* okazali pomomoÄb‫״‬ v dele izvra & b e n ij. “ V. Baza- 1iteraturovedenie za 50 l e t , Leningrad 21. Pospelov, turovedenija , denie - L ite r a - die v u l 9 garno-sociologi&eskich 82) habe w ird, preodolenija S. b e f r e it "v u lg ä r-s o z io lo - damit bemerkt: 1ite ra tu ro ve d e n iju ( i . d r.), habe, Voraus- sowjetischen is t sovetskomu 1968, gemacht ЗОег 5. Dement'ev ‫״‬KommunistiÖeskaja nov koto- Anfang der Realismus". P a rte i gewürdigt der Mnogoobrazie beskogo realizme, ta Gedankenduktus bemüht: Tom I I. 80) i kanonov rappovskogo p e rio d a ." • 0 RAPP überwunden habe. 78) 81) 20er, daß man sich gemeint sehe Literaturw issenschaft 60er "... [is b e z a e t], rappovcev prodolfcalas‫ ״‬nedolgo"e i , die daß F o r ts c h r itte - wie der gewesen, schem i E ntstellungen"•1 konnten, Ende s o z ia lis tis c h e n daß mit lite ra tu r th e o r e tis c h e tefcenij** gewaltige dem W ege des dogmatibeskich Bazanov daß Sowjet 1it e r a t u r “ auf fe s t, rappovcy. 9 7‫״‬ k o n s ta tie rt, und zwar Kovalev " tvorbeskich kul ‫ ״‬t iv ir o v a l i Asadullaev Jahre bor'bo s te llt in : Metodologibeskoe r a z v it ie V. za p j a t ’ desjat Kule&ov le t, ( iz d . ) , sovetskogo Sovetskoe lit e r a - 1ite ra tu ro v e - Diether Götz - 9783954791927 Moskva 1967, S. 51• Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 1. In 2. der sowjetischen L ite r a tu r setzt setzesmäßige und k la re Entwicklung z ia lis tis c h e n Realismus ein. Die des Methode der schaft e in z ig die hin s o z ia lis tis c h e n Bedingungen sich zur an eine ge- Methode des so- Realismus entwickelnden mögliche von Anfang is t unter s o z ia lis tis c h e n und r ic h t ig e den G esell- Methode in der Kunst3. Die Partei h ilf t und Durchsetzung 4. Die den Kunstschaffenden dieser S tr e itig k e ite n etwa im eher l i s t i sehen eine sie den einzelnen Bereich geschadet temporäre tenden zwischen Realismus haben der und die Entwicklung Erscheinung, ln der Organisât ionen Entwicklung Hinentwicklung behindert, Klassenkampf der Methode. 1ite ra ris c h e n L ite r a tu r bei zum sozia- doch waren sie auch Ausdruck Kunst; le tz tlic h vom noch anhal- k e in e s fa lls aber beim le tz te n Punkt wird d e u tlic h , daß die der 60er Jahre die V ie lfa lt Gruppierungen der 20er Jahre zweitrangig sehe Literaturw issenschaft terarischen Bedeutung te ra tu r und negativ fUr die Gesamtentwicklung der UUrdigung 1925 anzutreffen, sie wiederholt der Gesamtbewertung der RAPP-Periode. Die sowjetische sehen den je w e ils chen Elementen der der RAPP, was h in d e rt, des sie verschiedenen einer die was fö rd e rt n ic h t sich s e it die o ft den 30er knappen, nur was n ic h t in daß und Bewertung der äußert gnUgt: ihre s o z ia lis tis c h e n Wissenschaft d if f e r e n z ie r t die von 1925 bzw. Sehweise nur v o lle Entwicklung intensive RAPP in der hinein die noch mit wenigen und gewichtiger, frUhen mehr sein zwi- in h a ltli- Herrschaft die Frage: wird Dar- L ite ra tu i— 50er weichen in d ire kte n d.h. L it e r a - angelegte verkürzenden Darstellung bei und Durchset- sowjetischen in sich diesem Aspekt und b r e it bedeutsam und gewichtig bedeutsamer der noch auf Realismus? Unter b is n ic h t Voraussetzungen und ZK-Resolution verengt ve rstän dlich, Stellung Jahre ih r e r S ow jetiІ - ZK-Resolution 60er bei li- der der der war b e re its der in diese turwissenschaft Tendenz s o w je ti- d a r s te llt; es seien Uberzubewerten. Gerade zung der Jahre mußte, Hinweisen s o ll, s o ll a u s fü h rlich e r beauch Ueise Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 36 geschildert werden. Daß diese sich st immte r 1i t e ra tu rpol i t i scher hig in te rp re ta tio n s b e d U rftig und meinpoli t ischen, Wicklungen die im sowjetische tages ein konkret, Entwicklungen zu Sowjetstaat, s o ll ParteifUhrung hatte starkes Interesse o ffe n s ic h tlic h e n erscheinen den zu lassen, Charakter diesem von ta to risch e lismus und selbst die nicht werden; Sicht Ende die Erwähnt lung mus stark an die te rh in , P artei is t, der der vennych RAPP, 04. der aus an deren Realismus von dieser Literaturw issenschaft gebunden wird. erlassen Datum, dann eine d ire kte her darzustellen sein. bei der wurde, lite ra r is c h e Resolution wird hätte hin Verbindung Bedeutsam i s t wei- an dem mit H ilf e Realismus jener Tag, der gelungen an dem die 1iteraturno-chudoüestmit der vor allem die aufgelöst S c h rifts te lle rk o n g re ß gezogen. der RealisJahre Dies wird noch nä- RAPP durch die Literaturw issenschaft s e it s tie r t der m itg lie d e rs tä rk s te die der Organisationen zum I. Beurteilung Entwick- 60er tische daß sie v o lle n s o z ia lis tis c h e n "О perestrojke auch andere RAPP, hätten können, Prozeß der 1932 angesehen wird, organized j " auf Gruppierungen auslösen Methode des das entscheidende ZK-Resolution Ulas f ä l l t Er- Programmatik Durchbruch zum s o z ia lis tis c h e n aber wurden; lite ra r is c h e n daß der Person G o r 'k ijs 23. berUhmte der sowjetischen daß als denen d ik - zu deutliche des s o z ia lis tis c h e n werden m uß noch, der in können. und Durchsetzung in ausführlichen Rea- g e s te llt Diskussionen Unter s o z ia lis tis c h e n der geraten mit eher des und deren verschiedenartige Gefahr Instrument Ande- Jahre in a ls Raum zu geben. 20er V e rb indlichkeit v e r b in d li- v ie l an die Partei vor zu verleihen. Sowjet 1i t e r a t u r , a llz u der P a rte i- von S c h r if t s t e lle r n opportun, inrierungen der Vielzahl werden: des XX. n ic h t L ib e r a lit ä t V erb in dlich ke it Frage a llg e - gesamten K u l t u r p o li t i k war es nicht in angedeutet nach wie Realismus ih re r jenen Phasen der s o llt e die der Vorgänge und Ent- im Gefolge D is z ip lin ie ru n g ZUge vorherrschten, r e r s e its nur be- in te r p r e ta t ionsf ä- h in s ic h t lic h h ie r daran, r e la t iv e r Gesichtspunkt Darstellungen sind Darstellungen innenpolitischen che Methode des s o z ia lis tis c h e n der ändernden den 70er Jahren? Er&ov sowjea tte - L ite ra tu ro rg a - Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access n is a tio n merhin der so hätten. sistēmu pereme&ivalis“ vernye digen gerufen Weltanschauung im Das Ästhetik zu s c h e ite rt n ic h t in nur der Bemühen aber einen Rolle dann auch das Uber her n ic h t RAPP aber aufgrund seines is t: “ Odnako, e s li to Vychodcev RAPP durch bto s ta li v Dement'ev der RAPP 70er rung", in Zwar ih r e r die der Er&ov, Is t o r ija ..., S. Er&ov, Is t o r ija ..., S. 1979, S. A. vom vsesojuznom godov, Dement'ev, Voprosy s-ezde paßt das der der vom Bewer- 1932 Auflösung le d ig lic h organizacionnye o&ibki das RAPP klingenden der davon, ramki RAPP Ton schlägt lin ii RAPP vse 1 nedostatki. " 4 die der T ä tig k e it DD R in RAPP "terminologische den Verwir- und Organi- 44. 44. russkoj sovetskoj socialistibeskogo sovetskich iskusstvovedenija Moskva Diskussion lite r a tu r y , 242f. 86) sovetskogo Is t o r ija auch bemerkenswert zu anderen S c h r if t s t e lle r n L. Moskva sondern in Bewertung e rfä h rt L. Vychodcev, daß RAPP d e ja te l’ nosti Literaturw issenschaft werden ge- Darstellung llte r a tu r n o - p o liti& e s k o j Haltung a ls a ne poz i t i v noe. 4 •‫״‬ obstanovke 84) P. damit, ausübte, r e z u l't a t y der neuen aufgegeben werden - , vom A p r il 83) 85) er K o n tin u itä t n ich t s e r’ eznejttie auch durch einer le tz te n d lic h im Zusammenhang mit d iffe r e n z ie r te Jahren. die negativnoe, v negativ uporstvo) abwägenden Charakters ZK-Beschluß "... ähnlich - der Richtungen heraus- d i f ferenzierende Den noch am schärfsten an: i Als kunsttheoretischen ocenivat‫״‬ s i l ’ nee obozna&alis* Eine eher izmeniv&ejsja uzkim i. " 1* W ic h tig k e it RAPP abgegebene Gesamturteil, s p ric h t den wo etwa Grundlagen c u ltu r e l" ohne Not prevy&aet Auch "... kann bewertet, wo nach dem "leben- k o n s ta tie rt Überrascht von p r o t i vore£ivo hervorhebt. Er&ov "Programma p o s it iv RAPP die RAPP, sieht diese die tung celom, der im- Als (n a sto j& ivo st‫״‬ überhaupt In kotorych lite ra ris c h e n “ terrorism e s p ie lte. Ergebnis wo sich Furmanov oder dann f o r t : gefordert, immerhin lit e r a t u r - , eine v anderen Reihen s ofciboünymi. “ * ‫נ‬ wird, form ulieren, an, fä h rt RAPP-Theoretik e r Intoleranz RAPP-Organe gegenüber deren wie Fadeev, Werk des Künstlers hartnäckige g e s t e llt. in sredi polo¥enija den K lassikern“ Menschen" die Er tezisov, Jene Äußerungen der "das Lernen bei wird gewesen sei, S c h r i f t s t e ll e r befunden v k lju b a la werden Jahre bekannte Serafimovib RAPP 20er 1977, S. 30. І realizma p is a t e le j, èsteti&eskoj in : Iz mysli na peris to r i! 1930-ch Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 38 sationen starres sches“ Denken Gork i j als Freund-Feind-Denken vorgeworfen, Zeuge genannt, Auseinandersetzung mit te, an Bevor sie die sie durch einen wird der In ih re r im Zusammenhang mit einmal ZK-Resolution fe s t: den vor er fe n .“ ■• Bei anderer den 20er s c h r it t - Vereinigung Begeisterung' B lic k bekannte der p o litis c h e in s te llte erwähnt, RAPP z i e lt e : werden, die Helden Hinwendung der v e r tie fte n der die u r te ilt , d e u tlic h T ä tig k e it Als aber dieses d i f fe re n z ie rte re r g le ic h la u te n (s. a ls wird tu r d ik ta tu r angestrebt 87) N. S. 88) W . Beitz, daß die daß die der in F o rtdes Fordeso etwa jüngere der RAPP-Leute eine hätten, L it e - Beurteilung anlegt: negativ pauschaler, zwar wird und ausgeübt in einem zusammenschauen- überwiegend n ich t Weise g e f ä l l t , aufgegriffen, 85). Gestal- ein RAPP genannt, D D R in RAPP insgesamt U r t e il Anm. dieser anderen Beurteilungsmaßstab sichtspunkt Thun, - V e rtre te r unübersehbar, UdSSR und der der schaf- Sowjet l i t e r a t u r Klassiker Aussagen unter so i s t anschwellen- Bahn zu der wird zu eng ge- psychologischen Averbach. man diese indes re a l i s t І sehen Darstellung bezeichnet. allem f r e ie RAPP genannt Lind wird “ Der um dem mächtig Repräsentanten der einen betrieben, Beschluß auf dem Vorbild der hätte doch die d iffe re n z ie rte n raturwissenschaft sie obwohl namentlich werden führende RAPP gemacht, mehr zu einer Gesichtspunkt, Sowjet l i - 1932 wird vor habe P a r t e ilic h k e it Verunglimpfung vom "engen Rahmen" der sei, russischen Vorwurf versäum- zu erinnern: v e r fa lle n und echte a ls schärfsten n ic h t bestehenden Organi sa tionen war nach Betrachtet in RAPP . .. immerhin noch bei Jahren Bezymenskij den der Vychodcev wird 1iterarischen rung Z e it Schöpfertums S te lle tungsweise der s o z ia lis tis c h e n den Formulierung An Geringerer der “ Geschichte mußte gesprengt Strom kein dem ZK-Beschluß vom A p ril allem Der Rahmen der worden, der "v u lg ä rs o z io lo g i- wird in dieser klaren L ite r a tu r k r it ik nicht der Leitung RAPP der in RAPP der “ adm inistrative ausgezeichnet.“ • 7 te r a tu r “ “. .. großen Verdienste in sich jedoch und aber der Zwar be- sondern bekannte Ge- Art L ite r a - ih r Beitrag 76f. Die Lenin‫״‬sehe K u lt u r p o lit ik ..., S. 553. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 39 zuг lite ra tu rth e o re tis c h e n p o s itiv d a r g e s te llt; früheren Bewertungen der ser dies Sehweise e rö ffn e t vorwiegend n ich t p o s it iv in W enn also die vor allem lebendigen Menschen“ L ite r a tu r schaft so Analyse is t in Realismus sich Länder als der daß die die sich p o s itiv auf die wenn sie wäre. "nach dem herausge- immer häufiger an- Literaturw issenwonach der so- Methode aufzufassen interessanter Jahre RAPP auch von b e rü c k s ic h tig t, 20er d ie - Inhalte gegenwärt igen ein Bei bei 1ite ra tu rth e o - Forderung den der v e rfa lle n der a ls lite r a r is c h e h ie r Sowjet 1it e r a t u r fe h lt. ih re L ite r a tu r , der v ö llig werden können, und man andererseits in Aspekt, von RAPP in Forderung also, bedeutend und jener Begeisterung" Sonderheit der als M öglichkeit, b e u r t e ilt in Standpunkt zeichnet der die Beitrag s o z ia lis tis c h e r z ia lis t is c h e sei, eine bezieht, zutreffenden sich der wird RAPP-Tätigkeit “ adm inistrative Diskussion, wird, gerade hatte retischen s te llt Diskussion Wandel in der ab. Diether Götz - 9783954791927 Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 2.2.4. Der A m 23. Uber ZK-Beschluß vom 23. A p r il die 1932 e rlie ß “ Umbildung der ( “0 perestrojke Oie in sich in wicklung im ersten "... bto k o libestvennyj, ku sstva.H,° gen die NEP-Zeit, 1er “ noch tak zu schaffen, S c h r i f t s t e l 1er seien, so und waren“ “ ... neue Kolchosen eng4 ' genannt; dies in ein vor le rn , H. .. fo rt, namentlich die r e n . “ 92 Die gung sind a lle r mit Lage sehe Fraktion H inter Zeichnung sojuznaja nennung 90) A. v pebati, die hat, analoge der in Zwischenzeit Aufgaben der Anm. d. auf es aus der Bereich (!) - in eine in kommunisti- die damalige Sowjetunion ( bol ‫״‬î&evik i ) “ ; i V e re in i- anderen Bereichen dem XIX. d ir e k tiv y her- Sehri f t s te 11er sich P a r tija sympathisie- RAPP und VOAPP, auch lo s - und KUnst- und die der Osnovnye RAPM Gegenwart Sozialismus Partei S. RAPP, und v e rb irg t 1937, “ Rahmen der v e rfie le n kommunistischen erst aus Be- Z irk e lle b e n “ AbkUrzung VKP(b) FogeleviÜ, Kunstkader 0 r- Maßnahmen auch e r fo lg t der daß diese der Moskva der Komponisten, der KPSS in sich, der - der und KUnst- werden VOAPP, unterstützenden Verband, in geholfen und KUnstler das ZK e rlä ß t Auflösung Kommunist ibeska ja in In e in i- p r o le ta r ischen im 1ite ra ris c h -k U n s t1erisehen Sowjetmacht einem e in h e itlic h e n der is - und Kunstorganis a tionen Gefahr dem Aufbau des folgende: die h ie r von Jenen S c h r if t s t e lle r n Maßnahmen, fe s tg e s te llte n leitet, aber allem so daß der p ro le ta ris c h e r berge vor i "vor p ro le ta ris c h e L it e r a t u r - ‫״‬abgekapseltes allem die Position zu s tä rk e n ."91 zunehmend von den p o litis c h e n lösten, 89) - Assoziation ganisationen sich sei daß Sehri f t s t e 11er nachgewachsen, b e re its Ve r f . ] l it e r a t u r y P artei S c h r if t s t e lle r proletarischen [Russische die b o l'Ä o j S c h rif t s t e l l e r und die p r o le ta r ischer vorhandenen zu ro s t Ent- grundsätz 1ich dostignut es dann w eiter, Verordnung und ... g lie d e r t bisherige und “ fremden Elementen“ , schwach die gody organizaci j " ). die eingegangen p ro le ta ris c h e r um damit fä h rt Verordnung Verordnung auf kebestvennyj und KUnstler herangewachsen trie b en i mit Organisat ionen wird za poślednie Kader Auseinandersetzung habe. T e il Einschränkend heißt Jahren4 ‫״‬ VKP(b)9 die knapp gehaltene Sowjet 1it e r a t u r f e s t g e s t e llt , как das ZK der 1iteraturno-chudoìÉestvennych Teile: der 1932 Literatur— und Kunstorgarnis a tionen" ihrem Umfang eher zwei 04. Be- “ Vse- die Umbe- P arteitag 1952. zakonodatel9stva 50. 91) A. Fogelevib, S. 50, s. 92) A. Fogelevib, S. 50, s. Anhang N r.6. Diether Götz - 9783954791927 Anhang N r.7. Downloaded from PubFactory at 01/10/2019 03:46:08AM via free access 00050394 41 des Kunstschaffens * ,praktische dieser Die Bereich überhaupt rischen e rlä ß t mit Bereich Verordnung ra tu r- als nämlich mit der die Stellung der P artei nen und deren Auflösung solcher bestimmten des tu rp o litisch e m eine setzen. is t die es der á È Die Vorgaben liehen dadurch, (noch) fü r ta k tis c h e r n ö tig Bezug. sind daß vom künftigen Sozialismus verlangt Fogelevib, S. 50, allgemein sympathisieren“ , war, zu und zu u n te r- Natur: wenn es auf die Bildung so gebietet auf es ein lite r a - Organisation zu LiteraturorganisatIonen Diskussion zwischen Dies Bereich Probleme den e in z e l- Bereich erscheint zunächst einmal nimmt auch e in - darum geht, organisatorisch lit e r a r is c h e r die In h a lte gefragt.	14,941
https://openalex.org/W4387338738	OpenAlex	Open Science	CC-By	2,023	The contributing role of family, religious activities and school in protecting the mental well-being of school going adolescents in a rural district of Sri Lanka	N. H. S. Nadeeka	English	Spoken	4,431	9,193	Abstract Introduction: Mental health problems in adolescence are common. Introduction: Mental health problems in adolescence are common. Objectives: To assess the mental health of grade 10 students in Ampara District and their associated factors Objectives: To assess the mental health of grade 10 students in Ampara District and their associated fa Methods: This descriptive study was conducted from May to November 2018. Strengths and Difficulties Questionnaire (SDQ) was used to assess the students’ mental health status under five domains. They were total abnormal scores (TAS), emotional problems (EP), conduct problems (CP), hyperactive/inattention problems (HIP) and peer problems (PP). The prevalence of mental health problems was summarized as percentages. The significance of associated factors was assessed at p<0.05 and presented in adjusted odds ratio (aOR). Results: Of the 1340 students, 11.9% had reported TAS. The prevalence of EP, CP, PP and HIP were 15.1%, 12.9%, 12.1% and 11.7%, respectively. Eight factors associated with abnormal scores were identified. Difficulties in schoolwork were associated with all five domains (aOR=5.9; 95% CI: 2.5, 13.6 for TAS; aOR=4.0; 95% CI: 2.1, 7.7 for EP, aOR=2.9; 95% CI: 1.6, 5.4 for CP, aOR =2.6; 95% CI: 1.4, 4.6 for PP and aOR=2.5; 95% CI: 1.4, 4.5 for HIP). Not involving in religious activities regularly at home was associated with all domains except emotional problems (aOR=2.4; 95% CI: 1.3, 4.3 for TAS, aOR=2.2; 95% CI: 1.2, 3.9 for PP, aOR=2.1; 95% CI: 1.2, 3.6 for HIP and aOR=1.9; 95% CI: 1.2, 3.5 for CP). Not having a close family member to discuss their problems was associated with TAS (aOR=2.4; 95% CI: 1.4, 4.2), EP (aOR=3.4; 95% CI: 1.9, 5.8) and HIP (aOR=2.2; 95% CI: 1.3, 3.7). The presence of stressful life events was associated with TAS (aOR= 1.9; 95% CI: 1.1, 3.4), EP (aOR=1.9; 95% CI: 1.1, 3.5) and CP (aOR=2.3; 95% CI: 1.3, 4.3). Tobacco smoking habit of father was associated with TAS (aOR=2.9; 95% CI: 1.2, 6.6) and CP (aOR=2.3; 95% CI: 1.1, 4.6). The alcohol-drinking habit of father was associated with EP (aOR=3.0; 95% CI: 1.3, 7.0). Not attending to additional tuition classes was associated with EP (aOR=1.9; 95% CI: 1.1, 3.4) and PP (aOR=2.6; 95% CI: 1.4, 4.6). The low educational level of the mother was associated with the TAS (aOR=1.9; 95% CI: 1.2, 3.4). Conclusions & Recommendations: Mental health problems are common among school-going adolescents. Original Research NHS Nadeeka* & Kumudu Wijewardena Received on 8 May 2023 Accepted on 3 July 2023 Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) The contributing role of family, religious activities and school in protecting the mental well-being of school going adolescents in a rural district of Sri Lanka NHS Nadeeka* & Kumudu Wijewardena Department of Community Medicine, Faculty of Medical Sciences, University of Sri Jayewardenepura, Sri Lanka Correspondence: sajaninadeeka@gmail.com DOI: https://doi.org/10.4038/jccpsl.v29i3.8608 Received on 8 May 2023 Accepted on 3 July 2023 https://orcid.org/0000-0003-1201-3514 Original Research Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) The contributing role of family, religious activities and school in protecting the mental well-being of school going adolescents in a rural district of Sri Lanka NHS Nadeeka & Kumudu Wijewardena Department of Community Medicine, Faculty of Medical Sciences, University of Sri Jayewardenepura, Sri Lanka *Correspondence: sajaninadeeka@gmail.com DOI: https://doi.org/10.4038/jccpsl.v29i3.8608 Received on 8 May 2023 Accepted on 3 July 2023 https://orcid.org/0000-0003-1201-3514 Original Research Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) Methods This study was conducted from May 2018 to March 2019 in Ampara Regional Director of Health Service (RDHS) area. All the schools in this area are Sinhala medium schools and belong to three educational zones, namely Ampara, Mahaoya and Dehiattakandiya. Ampara Education Zone consists of four educational divisions, namely Ampara, Uhana, Damana and Lahugala. Mahaoya Zone consists of Mahaoya and Padiyathalawa divisions. Dehiattakandiya Zone consists of Dehiattakandiya West and Dehiattakandiya East divisions. The sample size for the prevalence study was calculated using an appropriate formula (8). With a prevalence of 16.1% of mental health problems (9), 95% confidence interval, 3% precision, 2.1 design effect (10) and 10% non-response rate, it was taken as 1338. The sampling technique used was multistage stratified cluster sampling with probability proportionate to the size of study population of each educational zone in Ampara RDHS Area. In 2018, there were 4278 students in 118 schools with the student population varying from 141 to 1156 in education divisions, while the number of students in a class varied from 2 to 44. Considering the geographical distribution and variations in the student population, all educational divisions were included in the study. A class of grade 10 of type 1AB, 1C or II schools was defined as a cluster, with an average cluster size of 26. The list of schools that was used as the sampling frame was obtained from the planning unit of each zonal educational office and the RDHS Ampara and cross-checked. One school in Ampara Zone was excluded from the study as it did not belong to Ampara RDHS area. All the students of each selected cluster (class of grade 10) were included in the study. As the World Health Organization estimates, 10- 20% of children and adolescents worldwide experience mental disorders, and half of them, begin by the age of 14 years (1). According to a nation- wide survey, nearly one-fifth of Sri Lankan students had mental health problems (4). Adolescents’ mental health is associated with many biological, psychological and social factors. The more risk factors adolescents are exposed to, the greater the impact on their mental health. The protective environment of the family, school and community is extremely important to prevent mental health problems in adolescents. Adolescents with mental health problems are more prone to social exclusion, stigma, poor academic performances, risky behaviours and human rights violations. Abstract All mental health problems were significantly associated with difficulties in schoolwork and most of the others were modifiable family-related factors. School mental health screening programmes are recommended with special attention on students with difficulties in schoolwork. School Mental health awareness and promotional programmes should target the students as well as teachers and parents to create a supportive school and family environment. Keywords: adolescents, associated factors, mental health, SDQ Keywords: adolescents, associated factors, mental health, SDQ Journal of the College of Community Physicians of Sri Lanka 167 Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) their associated factors of school-going adolescents in a rural conflict-affected area of Sri Lanka. Introduction Adolescence is considered as the age from 10 to 19 years and is the transient period between childhood and adulthood. During this sensitive period, a child becomes an adult with intellectual, physical, hormonal and social changes. Therefore, it is a period of increased vulnerability to mental health problems related to the regulation of affect/emotions and behaviour (1). Childhood mental health problems can be categorized into three groups, namely emotional disorders, externalized disorders and developmental disorders. Anxiety, depression, excessive irritability, frustration and anger are some of the common emotional problems (2-3). Examples of behavioural disorders are disruptive disorders, defiant disorders, aggressive behaviours and antisocial behaviours (1-2). Journal of the College of Community Physicians of Sri Lanka Methods Therefore, mental health problems will affect adolescent's development, education, and social life (1, 5). Hence, identifying high-risk individuals, and detecting and treating their mental health problems are a priority. Ampara health district is situated in the Eastern Province of Sri Lanka. Its economy is mainly based on agriculture (6). The area was badly affected by the long-lasted civil war (7). Therefore, this study was conducted to examine the mental health status and Further, to identify the risk factors of the mental Journal of the College of Community Physicians of Sri Lanka Journal of the College of Community Physicians of Sri Lanka 168 Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) together) groups and presented as percentages. To examine the risk factors for abnormal mental health, aOR was calculated using logistic regression models. Data were analysed using the Statistical Package for the Social Sciences (SPSS) version 21. health problems, a cross-sectional analytical study was conducted. The sample size calculated using an appropriate formula (11) was 166, however, since there were only 160 students identified with mental health problems in the prevalence study, they were matched with randomly selected 160 individuals who did not have mental health problems. Results Strengths Difficulties Questionnaire (SDQ) which has been translated to Sinhalese and validated in Sri Lanka was used as the screening tool (4). This tool can be used among 2-17-year-old persons and contains 25 items in five domains on EP, CP, HIP, PP and prosocial behaviour (a positive behavioural component which is not discussed in this study). The scores obtained for EP, CP, HIP and PP were added together to generate the TAS. The standard cut-off points given by the developers were taken to categorise the participants as normal, borderline and abnormal (12). In addition, a self-administered questionnaire was used to gather information on socio-demographic factors, personal, family and school related factors. Out of the 1382 students invited from all three educational zones, 1340 participated in the study (response rate was 97%). There were 608 boys (45.4%). The greater proportion was Buddhists (99.4%). Nearly one-third claimed that their parents were involved in farming-related occupations. Most were from nuclear families (71.3%) and living with both parents (90.7%). More than three quarters (78.9%) of students claimed that their family had enough income for their expenses. Students’ mean score was 9.6 (SD=4.7) for TAS, 2.6 (SD=1.8) for EP, 1.8 (SD=1.4) for CP, 3.4 (SD 1.9) for HIP and 1.8 (SD 1.5) for PP. More girls reported TAS (11.9%) and EP (16.9%), while boys had more CP (14.0%) and PP (14.8%). The proportion of students having HIP was the same for both sexes (11.7%) (Table 1). Journal of the College of Community Physicians of Sri Lanka 169 Journal of the College of Community Physicians of Sri Lanka Data analysis We dichotomized the results obtained for the total SDQ score and subscales as ‘normal’ and ‘abnormal’ (borderline and abnormal groups were combined Table 1: Distribution of the participants having abnormal scores according to total and subscales score and gender (N=1340) SDQ component Sex Abnormal score Normal score Total No. % No. % Total score Female 87 11.9 645 88.1 732 Male 73 12.0 535 88.0 608 Emotional problems Female 124 16.9 608 83.1 732 Male 80 13.2 528 86.8 608 Conduct problems Female 87 11.9 645 88.1 732 Male 85 14.0 523 86.0 608 Hyperactivity/inattention problems Female 86 11.7 646 88.3 732 Male 71 11.7 537 88.3 608 Peer problems Female 72 9.8 660 90.2 732 Male 90 14.8 518 85.2 608 Table 1: Distribution of the participants having abnormal scores according to total and subscales score and gender (N=1340) of the participants having abnormal scores according to total and subscales score and Distribution of the participants having abnormal scores according to total and subscales score a 1340) 169 Nadeeka NHS & Wijewardena K. Data analysis JCCPSL 2023, 29 (3) Table 2: Associated factors of the total abnormal SDQ scores and sub-scales among the students Status B SE Wald df Sig OR 95% CI Total abnormal score (TAS) Having difficulties in schoolwork 1.7 0.4 17.3 1 <0.001 5.9 2.5, 13.6 Not having a close family member to discuss their problems 0.8 0.2 9.2 1 0.002 2.4 1.4, 4.2 No regular religious activities 0.8 0.3 7.9 1 0.005 2.4 1.3, 4.3 Tobacco smoking habit of father 1.0 0.4 6.5 1 0.010 2.9 1.2, 6.6 Past stressful life events 0.6 0.2 5.8 1 0.016 1.9 1.1, 3.4 Low education level of the mother 0.6 0.2 5.8 1 0.016 1.9 1.2, 3.4 Emotional problems (EP) Having difficulties in schoolwork 1.3 0.3 17.7 1 <0.001 4.0 2.1, 7.7 Not having a close family member to discuss their problems 1.2 0.2 19.1 1 <0.001 3.4 1.9, 5.8 Alcohol drinking habit of father 1.1 0.4 6.5 1 0.010 3.0 1.3, 7.0 Past Stressful life events 0.6 0.2 4.9 1 0.026 1.9 1.1, 3.5 Not attending additional tuition classes 0.6 0.2 4.6 1 0.031 1.9 1.1, 3.4 Conduct problems (CP) Having difficulties in schoolwork 1.0 0.1 12.1 1 <0.001 2.9 1.6, 5.4 Past stressful life events 0.8 0.3 7.8 1 0.005 2.3 1.3, 4.3 Tobacco smoking habits of father 0.8 0.3 5.3 1 0.021 2.3 1.1, 4.6 No regular religious activities 0.6 0.2 6.0 1 0.014 1.9 1.2, 3.5 Hyperactive/Inattention problems (IHP) Having difficulties in schoolwork 0.9 0.3 8.8 1 0.003 2.5 1.4, 4.5 Not having a close family member to discuss their problems 0.7 0.2 8.3 1 0.004 2.2 1.3, 3.7 Not regular religious activities 0.7 0.2 7.1 1 0.007 2.1 1.2, 3.6 Peer problems (PP) Not attending additional tuition classes 0.9 0.2 10.0 1 0.002 2.6 1.4, 4.6 No regular religious activities 0.8 0.2 7.1 1 0.007 2.2 1.2, 3.9 Having difficulties in schoolwork 0.7 0.3 4.9 1 0.026 2.1 1.1, 3.9 OR=odds ratio Associated factors of the total abnormal SDQ scores and sub-scales among the students Sex of the controls and cases showed no significant difference in relation to mental health problems. Data analysis However, difficulties in school work, not involving in routine religious activities at home, not having a close family member to discuss their problems, past stressful life events, tobacco smoking habit of father, alcohol drinking habit of father (only for EP), not attending to additional tuition classes (only for EP and PP) and low educational level of the mother (only for TAS) were significantly associated with emotional and behavioural problems of the students (Table 2). The Negelkarke R2 values (0.374 for TAS, 0.295 for EP, 0.175 for CP, 0.135 for HIP and 0.124 for PP) and the Hosmer Lemeshow goodness of fit Chi-squared values (χ2=9.5; p=0.215 for TAS; χ2=5.4; p=0.6 for EP; χ2=0.4; p=0.9 for CP; χ2=6.6; p=0.1 for HIP; and χ2=1.21; p=0.8 for PP) for the models indicated that they were good fits. 170 Journal of the College of Community Physicians of Sri Lanka Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) described in previous studies (16). Religiosity helps to buffer against some types of stressors through improved self-esteem and coping skills to improve mental health (17). Discussion As the response rate was 97%, this study would represent the Sinhalese school-going adolescents in agriculturally based rural areas of Sri Lanka. Positive family relationship was assessed in the study and results suggested that not having a close family member to discuss their problems is associated with TAS (aOR=2.4), HIP (aOR=2.2) and EP (aOR=3.4). Positive family relationships had improved the mental health of adolescents in the United States (18), emphasizing the importance of family relationships regarding the mental health of the students. The prevalence of TAS was 11.9% and that of EP was 15.2%. These findings are not comparable with the previous national survey done in 2016, in which the students reported 8.5% felt lonely, 6.8% had attempted suicide and 38.5% were bullied (13). The prevalence of mental health problems among 13-15 year old students of Jaffna District was 21% (14), while in a national study conducted in 2004, it was 18.9% among school-going adolescents (4). However, the global prevalence of mental health problems in childhood and adolescence was 10-20%, which is comparable to the results of the current study (1). Smoking habit of the father was significantly associated with TAS (aOR=2.9) and CP (aOR=2.3). Alcohol drinking habit of the father was associated only with EP (aOR=3.0). Both maternal and paternal alcohol abuse are risk factors for adolescents’ mental health problems and maternal alcohol abuse is riskier (19). As in the Sri Lankan culture, female alcohol consumption is minimal and it has not been identified as a risk factor in our study (20). In other studies, it was shown that parental smoking predicts higher levels of externalized behavioural problems in adolescents. Biological and neurological effects of smoking may contribute to emotional and behavioural problems among adolescents (21-22). Stressful life events during the past six months were associated with TAS (aOR=1.9), EP (aOR=1.9) and CP (aOR=2.3). Negative life events had increased the risk of emotional and behavioural problems among the Chinese adolescents (23). The detrimental effects of cumulative stress on children's mental health have been described in a study conducted in Jaffna, Sri Lanka (24). In the present study, having difficulties in schoolwork is associated with TAS and all subscales. Not having additional support for academic activities were associated with EP (aOR=1.9) and PP (aOR=2.6). Discussion Having additional support for academic activities may have been beneficial in school performances and lacking that facility may have caused emotional and peer problems among the students. However, as this is a descriptive study, we could not explain the temporal association between mental health problems and difficulties in schoolwork. Mental health problems in childhood increase the risk of poor academic grades during adolescence (15). Hence, it is important to do a mental health assessment of the students especially who are having difficulties in schoolwork refer them to the mental health professionals. In the present study, having difficulties in schoolwork is associated with TAS and all subscales. Not having additional support for academic activities were associated with EP (aOR=1.9) and PP (aOR=2.6). Having additional support for academic activities may have been beneficial in school performances and lacking that facility may have caused emotional and peer problems among the students. However, as this is a descriptive study, we could not explain the temporal association between mental health problems and difficulties in schoolwork. Mental health problems in childhood increase the risk of poor academic grades during adolescence (15). Hence, it is important to do a mental health assessment of the students especially who are having difficulties in schoolwork refer them to the mental health professionals. Not being involved in religious activities regularly at home showed a significant association with TAS (aOR=2.4), CP (aOR=1.9), HIP (aOR=2.1) and PP (aOR=2.2). The importance of religious activities in the mental health promotion of school-going adolescents by improving religious morality, coping, respect for diversity and connectedness are well Parental education plays a key role in adolescents’ mental health. The current study had shown that mothers’ low education level increased the risk of TAS (aOR=1.9). In the international literature too, parents’ educational level, especially that of mothers had been identified as a crucial factor in child Journal of the College of Community Physicians of Sri Lanka Journal of the College of Community Physicians of Sri Lanka 171 Nadeeka NHS & Wijewardena K. JCCPSL 2023, 29 (3) identified as a risk factor, it should be highlighted among the teachers in view of identifying and referring them for mental health services. Further, the importance of positive family environment to prevent and promote mental health of the students has been highlighted in this study. development and their coping abilities in later life. References 1. WHO. Mental Health of Adolescents. World Health Organization, 2021. Available from: who.int/news- room/fact-sheets/detail/adolescent-mental-health. Accessed 5 August 2023. Discussion An increase in maternal educational level has improved the child’s mental health status (25). However, sex, screening time, leisure time activities, family income, child’s chronic illnesses or school culture were not associated with mental health problems in our study sample even though they had been proven to be associated in previously done studies locally and globally (9, 26-30). Public Health Implications Acknowledgements: We would like to acknowledge the students for participating in the study and the parents for consenting to their children to participate. We would like to thank the Ministry of Health and Ministry of Education for supporting us in conducting this study. • The study shows that prevalence of emotional and behavioural problems of school going adolescents is a public health problem in Sri Lanka. Author contributions SN and KW were involved in developing the study design, proposal writing, analysis of data and manuscript writing. SN was involved in data collection. KW supervised the overall project. • Most of their associated factors are modifiable and family related. • The existing school health system could be utilized the address the mental health related issues faced by the adolescents. Conclusions & Recommendations Emotional and behavioural problems are common among school-going adolescents especially among those with difficulties in schoolwork. Hence, it is important to screen and refer them if indicated to the relevant management facilities. Further, programmes should be planned and implemented to improve the mental health literacy among parents aiming alleviation of family related modifiable associated factors of mental health problems. The prevalence of total and all subclasses of mental health problems were more than 10% of the population, thus could be considered a public health problem in Sri Lanka. It seems that children have an unmet need for mental health services. 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https://openalex.org/W268886039_1	Spanish-Science-Pile	Open Science	Various open science	2,019	Diseño e implementación de una solución de administración de tráfico de red basada en DNS y chequeos de disponibilidad	None	Spanish	Spoken	7,264	13,793	Diseño e implementación de una solución de administración de tráfico de red basada en DNS y chequeos de disponibilidad. Tesis para obtener el grado de Magister en Redes de Datos Universidad Nacional de La Plata Facultad de Informática Autor: Nicolás del Río Directora: Mg. Lía Molinari Co - director: Ing. Luis Marrone Noviembre de 2015 El presente trabajo ha sido presentado y expuesto por el tesista en el XX Congreso Argentino de Ciencias de la Computación, en el marco del IX Workshop Arquitectura, Redes y Sistemas Operativos (WARSO) realizado en la Universidad Nacional de La Matanza. Acceso al artículo: http://hdl.handle.net/10915/42219 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ …a mamá y papá … a los amores de mi vida: Mariana, Agus, Santi y Emi Página 1 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Agradecimientos Quiero agradecer a la Profesora Lía Molinari por haberme guiado no sólo en este trabajo final de Tesis, sino a lo largo de toda mi carrera de Grado y Postgrado. Gracias Lía por el apoyo constante. Al Profesor Luis Marrone por los consejos y conocimiento aportado para realizar este trabajo. A mis compañeros de trabajo y colegas que aportaron desde un mate hasta una buena idea para llevar a cabo la tarea. Por último y no menos importante, a mi familia por su apoyo constante. Gracias por bancarme tantas horas frente a la compu. Página 2 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Índice general Capítulo 1 – Introducción ……………………………………………………………………………………………………………………. 1.1 Motivación ………………………………………………………………………………………………………………. 1.2 Objetivos Propuestos ………………………………………………………………………………………………. Capítulo 2 – BGP …………………………………………………………………………………………………………………………………. 2.1 Introducción ……………………………………………………………………………………………………………. 2.2 Características …………………………………………………………………………………………………………. 2.3 Interconexión de redes ……………………………………………………………………………………………. 2.4 Estructura de InterNet ……………………………………………………………………………………………… 2.5 Sistemas Autónomos ……………………………………………………………………………………………….. 2.5.1 Características ……………………………………………………………………………………………. 2.5.2 Clasificación ……………………………………………………………………………………………….. 2.5.3 Requerimientos para obtener un bloque de Direcciones IP ………………………… 2.5.4 Requerimientos para obtener un número de Sistema Autónomo ………………. 2.6 Actualizaciones de rutas y Penalizaciones ……………………………………………………………….. 2.7 Conclusiones finales del capítulo ……………………………………………………………………………… Capítulo 3 – DNS …………………………………………………………………………………………………………………………………. 3.1 Introducción ……………………………………………………………………………………………………………. 3.2 Características y funcionamiento …………………………………………………………………………….. 3.3 Espacio de nombres …………………………………………………………………………………………………. 3.4 Delegación de zonas ………………………………………………………………………………………………… 3.5 Servidores DNS y zonas ……………………………………………………………………………………………. 3.6 Tipos de registros …………………………………………………………………………………………………….. 3.7 Proceso de resolución ……………………………………………………………………………………………… 3.8 Caching ……………………………………………………………………………………………………………………. 3.9 Actualización de Zonas …………………………………………………………………………………………….. 3.10 Extensiones de Seguridad DNSSEC …………………………………………………………………………… 3.11 Conclusiones finales del capítulo ……………………………………………………………………………… Capítulo 4 - Sistemas de Monitoreo …………………………………………………………………………………………………….. 4.1 Introducción ……………………………………………………………………………………………………………. 4.2 Características y funcionamiento …………………………………………………………………………….. 4.3 Arquitectura ……………………………………………………………………………………………………………. 4.4 Principios de monitoreo en Icinga ……………………………………………………………………………. 4.5 Configuración de Icinga ……………………………………………………………………………………………. 4.6 Interfaces de Acceso ………………………………………………………………………………………………… 4.7 Conclusiones finales del capítulo ……………………………………………………………………………… Capítulo 5 - Propuesta de administración de tráfico basada en DNS ……………………………………………………. 5.1 Introducción ……………………………………………………………………………………………………………. 5.2 Arquitectura general de la solución planteada ………………………………………………………… 5.3 Proceso de instalación y configuración de componentes …………………………………………. 5.3.1 Sistema Operativo y entorno base ……………………………………………………………… 5.3.2 Módulo de monitoreo ………………………………………………………………………………… 5.3.3 Módulo de resolución de nombres …………………………………………………………….. 5.3.4 Módulo administrador ……………………………………………………………………………….. 5.3.5 Módulo de Interfaz gráfica …………………………………………………………………………. 5.3.6 Funcionalidad de la Interfaz Gráfica …………………………………………………………… 5.3.6.1 Interfaz de Login …………………………………………………………………………… 5.3.6.2 Dashboard Inicial (Estado General) ………………………………………………. Página 3 9 9 10 11 11 12 16 17 19 19 19 20 21 22 23 25 25 25 26 30 31 35 37 39 42 43 45 46 46 46 47 49 51 54 56 57 57 57 60 61 62 67 70 72 75 75 75 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ 5.3.6.3 Agregar Equipo …………………………………………………………………………….. 5.3.6.4 Administrar Equipos …………………………………………………………………….. 5.3.6.5 Logs ……………………………………………………………………………………………… 5.3.6.6 DNS ………………………………………………………………………………………………. 5.3.6.7 Usuarios ……………………………………………………………………………………….. 5.3.7 Interacción de componentes ……………………………………………………………………… 5.3.7.1 Agregar un host a través de la interfaz gráfica ……………………………… 5.3.7.2 Editar/Eliminar un host a través de la interfaz gráfica …………………… 5.3.7.3 Agregar una zona de DNS a través de la interfaz gráfica ……………….. 5.3.7.4 Eliminar una zona de DNS a través de la interfaz gráfica ………………. 5.3.7.5 Detección de la caída/recuperación de un servicio desde el módulo administrador ………………………………………………………………. 5.3.8 Distribución de la solución Traffic Manager ……………………………………………….. 5.4 Conclusiones finales del capítulo ……………………………………………………………………………... Capítulo 6 - Validación de la Solución y Esquema de Pruebas ……………………………………………………………… 6.1 Introducción ……………………………………………………………………………………………………………. 6.2 Software utilizado para la realización de pruebas ……………………………………………………. 6.3 Primer escenario de Pruebas: Tiempo de convergencia de BGP ………………………………. 6.4 Segundo escenario de Pruebas: Tiempo de convergencia de BGP con penalización ………………………………………………………………………………………………………. 6.5 Tercer escenario de Pruebas: Administración de tráfico de red utilizando DNS y chequeos de disponibilidad ………………………………………………………….. 6.6 Escenarios de Fallas controlados por la herramienta ……………………………………………………….. 6.7 Conclusiones finales del capítulo ……………………………………………………………………………… Capítulo 7 – Conclusiones ……………………………………………………………………………………………………………………. 7.1 Conclusiones finales ………………………………………………………………………………………………… 7.2 Trabajo a futuro ………………………………………………………………………………………………………. Anexo A - Arquitectura REST ……………………………………………………………………………………………………………….. Bibliografía …………………………………………………………………………………………………………………………………………. Página 4 76 78 79 79 80 81 82 83 84 85 86 87 88 89 89 89 90 100 107 124 125 126 126 127 128 130 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Índice de cuadros 3.5.1: Configuración de zona de DNS en servidor BIND …………………………………………..…………………………… 3.6.1: Ejemplo de definición de registros en zona de DNS …………………………………………………..………………. 3.8.1: Prototipo de las funciones gethostbyname y getaddrinfo …………………………………………………….…… 3.9.1: Ejemplo de ejecución del comando nsupdate en GNU/Linux …………………………………………………….. 3.9.2: Ejemplo de ejecución del comando nsupdate en GNU/Linux utilizando mecanismos de autenticación …………………………………………………………………………………………….. 3.10.1: Ejemplo de definición de zona que permite actualizaciones basadas en direcciones IP ……………………………………………………………………………………………………………………. 3.10.2: Ejemplo de definición de zona que permite actualizaciones basadas en certificados digitales …………………………………………………………………………………………………………. 4.4.1: Código de ejemplo de script para Icinga escrito en BASH ………………………………………………………….. 4.4.2: Definición de comando en Icinga ………………………………………………………………………………………………. 4.5.1: Definición de objeto Host en Icinga …………………………………………………………………………………………… 4.5.2: Definición de objeto CheckCommand en Icinga …………………………………………………………………………. 4.5.3: Definición de objeto EventCommand en Icinga …………………………………………………………………………. 4.5.4: Definición de objeto TimePeriod en Icinga ………………………………………………………………………………… 4.6.1: Formato genérico de requerimiento REST a Icinga ……………………………………………………………………. 4.6.2: Requerimiento REST de ejemplo a Icinga ………………………………………………………………………………….. 5.3.1.1: Software instalado en el sistema base ……………………………………………………………………………………. 5.3.1.2: Credenciales de usuario del Sistema Operativo Utilizado ……………………………………………………….. 5.3.2.1: Ejecución de comandos previos a la instalación de Icinga ………………………………………………………. 5.3.2.2: Ejecución de comandos para la instalación de Icinga ……………………………………………………………… 5.3.2.3: Ejecución de comandos para la instalación del motor de base de datos y la interfaz Icinga Web .. ……………………………………………………………………………………………… 5.3.2.4: Direcciones y credenciales de acceso a las interfaces Classic y Web de Icinga ………………………… 5.3.2.5: Credenciales de acceso a la interfaz REST de Icinga ………………………………………………………………… 5.3.2.6: Comandos para monitoreo definidos en Icinga ………………………………………………………………………. 5.3.2.7: Comando de Evento (EventCommand) definidos en Icinga …………………………………………………….. 5.3.3.1: Instalación de software de resolución de nombres BIND ………………………………………………………… 5.3.3.2: Generación de llaves con la herramienta dnssec-keygen ………………………………………………………… 5.3.3.3: Llaves creadas con la herramienta dnssec-keygen ………………………………………………………………….. 5.3.3.4: Definición de claves en servicio DNS para el servidor BIND …………………………………………………….. 5.3.3.5: Definición de zona de DNS en el servidor BIND gestionada por la herramienta Traffic Manager …………………………………………………………………………………………………….. 5.3.3.6: Definición zona de DNS en el servidor BIND con comando allow-transfer ………………………………. 5.3.3.7: Definición de clave y asociación al servidor primario en el servidor DNS secundario ……………… 5.3.3.8: Definición de zona de DNS en servidor secundario …………………………………………………………………. 6.3.1: Traslaciones de red (NAT) configurados para el primer escenario de pruebas …………………………… 6.3.2: Configuración del router R-ISP1 para el primer escenario de pruebas ……………………………………….. 6.3.3: Configuración del router R-AS6500 para el primer escenario de pruebas ………………………………….. 6.3.4: Configuración del router R-AS6501 para el primer escenario de pruebas ………………………………….. 6.3.5: Configuración del router R-AS6502 para el primer escenario de pruebas ………………………………….. 6.5.1: Traslaciones de red (NAT) configurados para el tercer escenario de pruebas en router R-ISP2 …………………………………………………………………………………………………………………….. 6.5.2: Traslaciones de red (NAT) reversos configurados para el tercer escenario de pruebas en los router R-ISP1 y R-ISP2 …………………………………………………………………………….. 6.5.3: Configuración del router R-ISP1 para el tercer escenario de pruebas ……………………………………….. 6.5.4: Configuración del router R-ISP2 para el tercer escenario de pruebas ……………………………………….. 6.5.5: Programa para la realización de chequeos en Windows para el Página 5 33 36 41 43 43 44 44 50 51 52 53 53 53 55 55 62 62 63 63 64 65 65 66 67 67 68 68 68 69 69 70 70 91 92 93 94 95 108 110 111 112 116 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ tercer escenario de pruebas ……………………………………………………………………………………………………………….. 6.5.6: Programa para la realización de chequeos en GNU/Linux para el tercer escenario de pruebas ………………………………………………………………………………………………………………. Página 6 119 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Índice de figuras 2.4.1: Estructura de la red InterNet en niveles ……………………………………………………………………………………. 2.5.2.1: Clasificación de sistemas autónomos ……………………………………………………………………………………… 2.6.1: Penalizaciones en BGP ………………………………………………………………………………………………………………. 3.2.1: Servidores DNS raíz …………………………………………………………………………………………………………………… 3.3.1: Estructura del sistema de nombres DNS en forma de árbol ………………………………………………………. 3.3.2: Inconsistencia de nombres en el sistema DNS …………………………………………………………………………… 3.4.1: Delegación de administración en una estructura de DNS ………………………………………………………….. 3.5.1: Esquema de dominios de DNS y delegación de zonas ………………………………………………………………… 3.7.1: Proceso de resolución de un nombre de DNS ……………………………………………………………………………. 4.3.1: Arquitectura de Icinga versión 2 ………………………………………………………………………………………………… 4.4.1: proceso de ejecución de chequeos a través de plugins en Icinga ………………………………………………. 4.5.1: Utilización de la interfaz icinga2 feature en Icinga …………………………………………………………………….. 4.6.1: Ejemplo de interfaz Classic Interface de Icinga ………………………………………………………………………….. 4.6.2: Ejemplo de interfaz Web Interface de Icinga ……………………………………………………………………………. 5.2.1: Componentes que forman parte de la solución Traffic Manager ………………………………………………. 5.3.5.1: Esquema de base de datos utilizado por la herramienta Traffic Manager ………………………………. 5.3.6.1.1: Pantalla de Login de la herramienta Traffic Manager ………………………………………………………….. 5.3.6.2.1: Pantalla Dashboard Inicia (estado general) de la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.6.3.1: Pantalla para Agregar Host en la herramienta Traffic Manager …………………………………………… 5.3.6.4.1: Pantalla de Administración de Hosts de la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.6.5.1: Pantalla de visualización de Logs de la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.6.6.1: Pantallas para la gestión de zonas de DNS en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.6.7.1: Pantallas para la gestión de usuarios en la herramienta Traffic Manager …………………………….. 5.3.6.7.2: Pantallas para la visualización de Logs de Auditoría en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.7.1.1: Diagrama de Flujo para el agregado de un Host en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.7.2.1: Diagrama de Flujo para Editar/Eliminar un Host en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.7.3.1: Diagrama de Flujo para el agregado de una zona de DNS en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.7.4.1: Diagrama de Flujo para eliminar una zona de DNS en la herramienta Traffic Manager ………………………………………………………………………………………………………………………………….. 5.3.7.5.1: Diagrama de Flujo para la detección/recuperación de un Host en la herramienta Traffic Manager ………………………………………………………………………………………………………………. 6.3.1: Topología de red utilizada en el primer escenario de prueba con BGP ………………………………………. 6.3.2: Prueba de ICMP desde la PC Cliente hacia la IP 190.40.66.11 ……………………………………………………. 6.3.3: Tabla de ruteo del router R-AS6501 ………………………………………………………………………………………….. 6.3.4: Tráfico ICMP desde la PC Cliente hacia la IP 19040.66.11 durante convergencia de BGP …………… 6.3.5: Detección de caída de vínculo por el proceso BGP en router R-AS6500 …………………………………….. 6.3.6: Detección de caída de vínculo por el proceso BGP en router R-AS6501 …………………………………….. 6.3.7: Tabla de ruteo del router R-AS6501 luego de la convergencia de BGP ………………………………………. 6.4.1: Configuración de penalizaciones de BGP en router R-AS6501 ……………………………………………………. 6.4.2: Tabla de ruteo de BGP en router R-AS6501 ……………………………………………………………………………….. 6.4.3: Tabla de ruteo de BGP en router R-AS6501 luego de detectar caída Página 7 18 20 23 26 27 28 31 32 38 48 49 52 54 55 58 74 75 76 78 78 79 80 81 81 82 83 84 85 86 91 96 97 98 99 99 100 101 102 103 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ de red 190.40.66.0/24 ………………………………………………………………………………………………………………………… 6.4.4: Análisis de penalidad para la red 190.40.66.0 en R-AS6501 ………………………………………………………. 6.4.5: Verificación de penalización de BGP por un camino para la red 190.40.66.0 en R-AS6501 ……………………………………………………………………………………………………………… 6.4.6: Penalización completa de BGP para la red 190.40.66.0 en R-AS6501 ………………………………………… 6.4.7: Supresión en la tabla de ruteo global de la ruta penalizada por BGP en R-AS6501 ……………………. 6.4.8: Verificación de pérdida de conectividad desde la PC Cliente hacia la IP 190.40.66.11 ……………….. 6.4.9: Despenalización de ruta en R-AS6501 ……………………………………………………………………………………….. 6.5.1: Topología de red utilizada en el tercer escenario de prueba con la herramienta Traffic Manager …………………………………………………………………………………………………………………………………… 6.5.2: Acceso desde la PC Cliente al servidor Web a través de R-ISP1. Direccionamiento afectado por el NAT ……………………………………………………………………………………………….. 6.5.3: Alta de Host www.ejemplo.com.ar en la herramienta Traffic Manager …………………………………….. 6.5.4: Resultado de la ejecución del programa creado para el tercer escenario de pruebas ………………………………………………………………………………………………………………………….. 6.5.5: Captura de tráfico realizada para el tercer escenario de pruebas ………………………………………………. 6.5.6: Visualización de Logs en la herramienta Traffic Manager para el tercer escenario de pruebas ………………………………………………………………………………………………………………………….. 6.5.7: Captura de tráfico DNS para el navegador Google Chrome 6.5.8: Cache de resolución de registros DNS del navegador Google Chrome (chrome://net-internals#dns) ……………………………………………………………………………………. 6.5.9: Captura de tráfico DNS para el navegador Internet Explorer …………………………………………………….. 6.5.10: Captura de tráfico DNS para el navegador Mozilla Firefox ………………………………………………………. Página 8 103 104 104 105 105 106 107 110 114 117 118 118 121 121 122 122 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Capítulo 1 Introducción 1.1 Motivación Los servicios de red como ser publicación de contenido WWW y mail entre otros, han crecido exponencialmente en los últimos años. El acceso a la red por parte de pequeñas organizaciones, e inclusive usuarios particulares, ha propiciado que la oferta de contenidos sea cada vez mayor, sin contar, en la gran mayoría de los casos, con grandes infraestructuras de hardware, redundancia y comunicaciones. Este crecimiento conlleva a la necesidad de contar con mecanismos de replicación, que garanticen alta disponibilidad. Las grandes organizaciones tienden a configurar sus servicios críticos en servidores redundantes y redes sin puntos únicos de falla con el fin de brindar elevados niveles de SLA1 (Service Level Agreement). Las tendencias actuales, llevan a los administradores a utilizar las técnicas anteriormente descriptas, así como también a desconsolidar los mismos en diversos centros de procesamiento de datos (CPD) con el fin de garantizar niveles mínimos de puntos únicos de falla. Históricamente se ha utilizado el protocolo Border Gateway Protocol (BGP), como única herramienta que permite conmutar el tráfico de red que está siendo dirigido a un centro de procesamiento de datos (CPD) hacia otra locación ante eventuales fallas de red. En la actualidad, contar con un bloque de direcciones IP propio, así como también un Número de Sistema Autónomo (ASN) de BGP en Internet es muy complejo y costoso, y no cualquier usuario puede acceder a ello2. Es por esta razón, que surge la necesidad de contar con una alternativa de solución a la problemática planteada, que sea de mayor alcance para este conjunto de usuarios que no cuenta con una gran infraestructura de comunicaciones. El protocolo Domain Name System (DNS), ha sido y sigue siéndolo, un gran pilar de la red de comunicaciones de Internet. Es a través del cual se permite localizar a los recursos en esta inmensa nube de contenidos. Si se pudieran cambiar los registros de resolución de DNS en base al estado de la red, sería posible direccionar los requerimientos de los usuarios hacia los enlaces o servicios que se encuentren operativos. Esta técnica podría adoptarse como opción a la utilización de un protocolo de ruteo por parte de organizaciones que posean una menor infraestructura. Es por esta razón, que se puede utilizar a este protocolo como herramienta complementaria a los protocolos de ruteo tradicionales. El protocolo BGP, por 1 Acuerdo de nivel de servicio: http://www.sla-zone.co.uk 2 Requerimientos para obtener un número de sistema autónomo (ASN): https://www.arin.net/resources/request/asn.html Página 9 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ su naturaleza, permite identificar las fallas de red y converger por sus propios medios, garantizando un nuevo camino para el ruteo de los datos solicitados. Surge entonces la pregunta: ¿Qué le falta al protocolo DNS para aproximarse al funcionamiento de BGP? La respuesta puede parecer trivial pero no lo es. En principio es posible indicar que lo que le falta al protocolo DNS para aproximarse al funcionamiento de BGP, es una componente que se encargue de determinar los estados de la red, disparar la convergencia y garantizar la consistencia y estabilidad de la red. 1.2 Objetivos Propuestos A lo largo del presente trabajo se analizarán los protocolos BGP y DNS, así como también los servicios de monitoreo que dotarán al servicio de resolución de nombres de una herramienta que permita contestar a la pregunta planteada. Adicionalmente se documentará el desarrollo de la herramienta “Traffic Manager”, la cual permitirá gestionar servicios a través del protocolo DNS y basándose en chequeos de disponibilidad de los mismos, tomar decisiones de convergencia. Finalmente, se realizarán y documentarán las pruebas de performance de la herramienta implementada y se evaluarán los resultados con el fin de compararlos con los resultados de convergencia arrojados por el protocolo BGP de modo tal de llegar a establecer bajo qué circunstancias es conveniente utilizar la tecnología propuesta, en qué casos resulta más favorable la utilización de BGP y qué impacto puede producir la utilización de ambas tecnologías conjuntamente. Página 10 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Capítulo 2 BGP 2.1 Introducción La red InterNet, tiene sus orígenes en su predecesora red ARPANET, la cual nació con el fin de interconectar nodos distantes para que los mismos pudieran transferir información entre ellos. Dicha red, nació en el año 1969 y en poco más de 2 años ya contaba con 40 nodos. Entre los años 1974 y 1982, se crearon una gran cantidad de redes entre las que se destacaron TELNET en 1974 que fue la versión comercial de ARPANET; USENET en 1979 para la utilización del e-mail, BITNET en 1981 que unía universidades de Estados unidos utilizando protocolos de IBM y EUNET en 1982 que unía el Reino Unido con Escandinavia y Holanda. El Protocolo utilizado por las máquinas conectadas a ARPANET se llamó NCP (Network Control Protocol o Protocolo de Control de Red), quien con el tiempo y con el fin de brindar una mayor escalabilidad, dio paso al protocolo TCP/IP en el año 1982. TCP/IP está formado no por uno, sino por varios protocolos, siendo los más importantes el protocolo TCP (Transmission Control Protocol o Protocolo de Control de Transmisión) y el Protocolo IP (Internet Protocol o Protocolo de Internet). Con el fin de que los datos puedan viajar a través de una red TCP/IP, los mismos son divididos en fragmentos o paquetes de información que viajan de forma independiente y se ensamblan al final del proceso. Este concepto se aplica a las redes conmutadas por paquetes, que es el tipo de red que utiliza InterNet hoy en día. El protocolo IP brinda direccionamiento a los nodos de la red de modo tal que se pueda seguir un camino para que los paquetes lleguen hacia su destino. Para ello utiliza un esquema de direccionamiento y un protocolo para la toma de decisiones conocido como “protocolo de ruteo”. La versión actual del protocolo IP es la 6 (definida en la RFC 2460 [1]); aunque las más utilizada, y en la que este trabajo se enfocará, es la versión 4 (definida en la RFC 791 [2]). Con el fin de encaminar los paquetes, se define el concepto de conmutador o router, que opera en el nivel de red del modelo OSI3 examinando las cabeceras de los paquetes IP. Los mismos poseen interfaces conectadas a más de una red, y su función principal es pasar los datos de una red a otra. Los routers permiten interconectar tanto redes de área local (LANs o Local Area Networks) como redes de área extensa (WANs o Wide Area Networks). Otra clave importante en este esquema, son los protocolos de ruteo. Los mismos nutren a los routers de información indispensable para encaminar los datos. En las redes LAN, se utilizan protocolos de ruteo interior (iGP o interior Gateway Protocol) como RIP4 y OSPF5 entre otros; o bien 3 Modelo de interconexión de sistemas abiertos (ISO/IEC): www.iso.org 4 RIP: Protocolo de ruteo interno definido en las RFC 1723 [3] y 2453 [4] 5 OSPF: Protocolo de ruteo interno definido en la RFC 2328 [5] Página 11 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ ruteo estático. En redes del tipo WAN se utilizan protocolos de ruteo exterior (eGP o exterior Gateway Protocol). En este capítulo, se analizarán las características del protocolo BGP (Border Gateway Protocol). 2.2 Características Los protocolos de ruteo exterior como BGP, tienen como propósito intercambiar información de ruteo entre organizaciones, conocidas también como sistemas autónomos. El primer protocolo de ruteo exterior que se implementó fue Gateway to Gateway Protocol (GGP) en el año 1980 y documentado en la RFC 823[6]. Lo sucedió Exterior Gateway Protocol (EGP) en el año 1982 y estandarizado en la RFC 827[7], que luego fue reemplazada por la RFC 904[8] en 1984. A medida que la red InterNet crecía, una mayor cantidad de sistemas autónomos eran interconectados con lo cual se comenzaron a divisar inconvenientes en la performance de EGP. Adicionalmente, surgieron necesidades de encaminar el tráfico de red basándose en métricas diferentes a las utilizadas, para las cuáles el protocolo no había sido implementado. Fue en ese entonces que resultó necesario crear un nuevo protocolo de ruteo conocido como BGP, que se adapte a las necesidades de crecimiento de la red. En junio de 1989 se estandarizó la primera versión del protocolo en la RCF 1105[9] (BGP1). La primera versión del estándar definió muchos de los conceptos que hoy son utilizados por el actual protocolo BGP, como formato de los mensajes y mecanismo de comunicación entre pares. La versión actual de BGP es la 4 (BGP4) y se encuentra documentada en la RFC 1771[10] y RFC 4271[11] desde el año 1995 y 2006 respectivamente. BGP 4 es el protocolo de ruteo exterior que utiliza InterNet desde el año 1995. Las principales ventajas de la versión 4 sobre sus predecesoras fueron la posibilidad de manejar classless inter domain routing (CIDR o enrutamiento entre dominios sin clases) y agregación con el fin de disminuir el tamaño de las tablas de ruteo, conceptos que serán discutidos más adelante. BGP es un protocolo de ruteo del tipo path-vector o vector de caminos, lo que significa que cuando un router recibe información acerca de un destino, recibe la lista completa de nodos que se debe atravesar para alcanzarlo. A través de esta información se calcula el camino óptimo hacia cada punto de la red basando su decisión en la longitud del vector. Con el fin de ocultar los pormenores de cada red, BGP define el concepto de sistema autónomo como: Un conjunto de routers que operan bajo una única política de ruteo y definen una o más organizaciones. [12] La métrica de BGP se basa en la cantidad de saltos, es decir, la cantidad de sistemas autónomos que debe atravesar el paquete para llegar a la red destino. De este modo, la ruta que tenga la menor cantidad de saltos, será la elegida como mejor ruta. Una característica importante de BGP, y lo que lo hace muy potente frente a otros protocolos, es la posibilidad de definir políticas de ruteo. Las políticas son implementadas en cada router, y permiten tomar decisiones diferentes a las que podrían haberse aprendido por el protocolo. Página 12 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ Mediante las políticas se puede manipular el ruteo y se pueden tomar decisiones acerca de dónde o hacia dónde transmitir el tráfico. Las políticas son implementadas a través de filtros, aceptando, rechazando o modificando rutas informadas por los vecinos de modo de manipular el flujo de los datos. BGP utiliza el puerto 179 de TCP para la comunicación con sus vecinos. Esto representa una gran diferencia respecto de otros protocolos, lo cuales corren sobre IP o UDP. Para que un router pueda intercambiar información con otros, primero debe establecer una sesión con uno o más vecinos, a través de la cual intercambiarán mensajes. Dicha sesión debe ser configurada manualmente en ambos extremos y permite mantener un link virtual entre 2 dispositivos, el cual permitirá no solo el intercambio de información, sino el monitoreo del estado de la red. En caso de detectarse una caída sobre la sesión configurada, se podrá inferir que el par está presentando inconvenientes, lo que generará que deban actualizarse las tablas de ruteo para dejar de enviar información a través de él. Cuando un router BGP se enciende, el mismo establece una sesión contra cada uno de sus vecinos configurados, lo que se conoce como “establecer una vecindad”, y envía los correspondientes saludos definidos en la RFC. Si la vecindad es aceptada, entonces se establece el emparejamiento o peering. Una vez establecida la vecindad/peering, ambos routers intercambian información, enviando una copia de sus propias tablas y recibiendo la correspondiente información de su par a través de mensajes del tipo “update”. En este punto es donde las políticas de ruteo cobran un rol importante, ya que en cualquier protocolo de ruteo estándar se enviaría toda la tabla de ruteo, mientras que en BGP se enviará la información que se encuentre definida en la política, ocurriendo lo mismo al incorporar las rutas recibidas a la tabla de ruteo. Una vez que este proceso se haya completado, los routers sólo intercambiarán mensajes del tipo “keepalive” para indicar que la sesión se encuentra activa y mensajes con la porción de la tabla de ruteo que haya sufrido cambio si es que lo hubiera. Con el fin de evitar procesamiento innecesario, las actualizaciones de las tablas de ruteo solo se enviarán al iniciarse un router o bien cuando se produzca algún cambio en la red. El protocolo BGP, define 4 tipos de mensajes que los routers pueden intercambiar con el fin de propagar la información de la red y mantener la misma consistente [13]: • OPEN: Una vez establecida la conexión entre 2 routers, es el primer mensaje de BGP que se envía. Su función principal es informar al vecino acerca de la versión del protocolo que se está utilizando y el número de sistema autónomo. Adicionalmente se enviará información acerca de la duración de la sesión, identificación del router que envía el mensaje; y en el caso de configurarse autenticación, se enviará información de la misma. • KEEPALIVE: Este tipo de mensaje, sirve para la confirmación del mensaje OPEN y para informar a los vecinos que la sesión se encuentra activa. Página 13 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ • NOTIFICATION: Permite cerrar la sesión, cerrando también la conexión TCP lo que causará que los vecinos deban modificar sus tablas de ruteo para dejar de enviar información a través del router que acaba de cerrar la sesión. • UPDATE: Este mensaje sirve para intercambiar información de ruteo y solo es enviado a los vecinos cuando ocurre algún cambio en la topología de red. El mismo contiene las rutas a agregar o eliminar, sus atributos y la longitud de cada una entre otras. La recepción por parte de un router de este mensaje implicará la modificación de sus tablas de ruteo, y que el mismo emita un nuevo mensaje UPDATE al resto de sus vecinos notificando los cambios. Todo esto, si la política local configurada lo define. Una de las características que hacen de BGP un protocolo potente, es la capacidad de tomar decisiones de ruteo basándose en diferentes métricas las cuales son descriptas por un conjunto de atributos y seleccionadas por una política de ruteo. Cuando un router envía información a otro a través de un mensaje UPDATE, la misma es descripta utilizando atributos. Dentro de los más importantes se pueden destacar[14]: • ORIGIN: Determina si la ruta fue aprendida mediante una actualización de BGP o de un protocolo de ruteo interno iGP. Las rutas pueden ser aprendidas por otros vecinos mediante mensajes UPDATE, o bien pueden ser aprendidas a través del protocolo de ruteo interno que utilice el sistema autónomo para el manejo de rutas dentro de la organización. Este atributo permite definir prioridades al momento de seleccionar una ruta, dándole mayor prioridad a una aprendida a través de iGP y luego será preferible una aprendida por eGP (Exterior Gateway Protocol, que en este caso será BGP) • AS_PATH: Contiene una lista de todos los números de sistema autónomo que debe atravesar el router para llegar al destino indicado. Este atributo tiene una especial importancia, ya que permite romper ciclos infinitos en la red (loops) y elegir la mejor ruta. Cada sistema autónomo agrega su número de ASN en este atributo para cada una de las rutas que aprende, antes de reenviarlas. De este modo se forma el camino utilizado por los routers para tomar las decisiones • NEXT-HOP: Cuando un router BGP anuncia una ruta a un vecino, utiliza este atributo para indicar cuál es la dirección IP del router que se deberá utilizar para alcanzar dicho destino. Este atributo es útil, en caso de que el destino que se deba utilizar no sea el mismo router que notificó la ruta por BGP. • LOCAL_PREF: Es un atributo interno definido por cada router que permite ponderar una ruta aprendida por iGP sobre otra aprendida por eGP. En caso de que la misma ruta haya sido aprendida por 2 caminos distintos, mediante este atributo se podrá definir cuál de ellas tendrá mayor prioridad sobre la otra. Página 14 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ • ATOMIC_AGREGATE: Este atributo permite indicar que la ruta en cuestión ha sido obtenida a través de la agregación de rutas más precisas. El mismo indica que las rutas han sido sumarizadas con el fin de disminuir la tabla de ruteo • MED (Multi-Exit-Discriminator): Este atributo tiene ámbito de aplicación entre sistemas autónomos vecinos, y se utiliza en caso de tener más de una entrada hacia un sistema autónomo con el fin de determinar hacia donde deben ser enviados los paquetes y brindar balanceo de carga en la red. Cuando un router recibe información de sus vecinos, lo primero que hará es verificar si su propia política de ruteo admite dicha información. En caso de no admitirla, la misma será descartada inmediatamente. Si la política acepta dicha información, entonces se verificará la nueva ruta respecto a las rutas que actualmente posee el dispositivo. Si la nueva ruta es considerada mejor que otra que actualmente existe, entonces se agregará la nueva ruta a la tabla de ruteo y se eliminará la antigua. Por último, si es que la política lo permite, se informará a los routers vecinos de dicha modificación para que los mismos repitan el procedimiento y la red pueda mantenerse en un estado coherente. El proceso de decisión de encaminamiento de BGP respecto a otros procesos de decisión es más complejo, ya que no solo verifica la dirección de red de destino sino que también toma decisiones en base a los atributos de cada una de las rutas aprendidas. Cuando un router BGP recibe un paquete para su encaminamiento, la primera acción que realizará es verificar la dirección de red destino del mismo con el fin de evaluar si en su tabla de ruteo posee una ruta que le permita llegar a dicha red. En caso de contar con más de una ruta, se seleccionará la menor de ellas utilizando el valor de la máscara de red definida en la tabla de ruteo. En caso de seguir teniendo varias alternativas para seleccionar, analizará el atributo LOCALPREF para dicha entrada y seleccionará la ruta que posea el número mayor. Si aún tuviera más de una ruta a dicho destino, analizará el atributo AS-PATH (lista de sistemas autónomos que debe atravesar) y seleccionará aquella ruta que posea un camino más corto. Para finalizar el proceso, si aún hubiera que tomar una decisión respecto a que ruta utilizar, se seleccionará como ruta con mayor preferencia a aquella que haya sido aprendida a través de iGP y por último aquella que posea un atributo Multi-Exit-Discriminator (MED) más bajo. Una vez seleccionada la ruta, el paquete será conmutado al próximo salto de la red para que el proceso continúe hasta alcanzar el destino. Página 15 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ 2.3 Interconexión de redes Con el fin de que las organizaciones puedan intercambiar información entre ellas, las mismas deben interconectarse entre sí utilizando tecnologías disponibles. La interconexión de redes define los pilares básicos de la arquitectura de la red InterNet. Cuando el volumen de datos que se debe transmitir desde una red hacia otra es bajo tiene sentido contratar el servicio de transporte a uno o más Proveedores de Servicios de InterNet (InterNet Service Provider, ISP). Este modo de conectividad es el que generalmente utilizan las organizaciones para unirse a la red. Cuando los volúmenes de información se incrementan hacia determinados puntos de la red, cobra sentido ampliar el esquema de interconexión, mediante la utilización de conexiones más complejas. Es en este punto donde nace la necesidad de realizar interconexiones con otras redes, con el fin de abaratar costos, ya que la métrica utilizada por los ISP para cobrar sus servicios, se basa en el volumen de tráfico que las organizaciones transfieren. La posibilidad de gestionar el tráfico de red independientemente de los ISP, devengará en un menor costo económico para las empresas. Existen diversas técnicas para la interconexión de redes. Desde el punto de vista técnico las configuraciones son similares. La principal diferencia entre ellas reside en cuestiones comerciales y económicas. La técnica más común es la contratación del servicio a un proveedor de servicios de InterNet. Mediante esta técnica, la organización contrata el servicio, el cuál será provisto por un único proveedor que será el responsable de transportar los datos de la organización hacia otras redes. Si la empresa no cuenta con direccionamiento de InterNet propio, el ISP le proporcionará un bloque de direcciones IP que estará asignado a la organización mientras dure el contrato con el mismo. Si la empresa cuenta con direccionamiento IP propio, entonces tendrá la opción de utilizarlo a través de la red del ISP. Para ello podrá optar por dialogar BGP directamente con otros routers, para lo cual además de poseer su propio bloque de direcciones IP tendrá que contar también con un número ASN propio; o bien podrá anunciar su bloque de direcciones IP a través del ASN del proveedor al que contrata el servicio. En esta configuración el proveedor cobrará a la organización el transporte de los datos basándose en cantidad total de tráfico transmitido, capacidad máxima de enlace que la organización necesitará y otros factores. A medida que el volumen de tráfico crece, la técnica anteriormente descripta puede resultar costosa para las organizaciones, con lo cual deberán evaluarse nuevas alternativas. Otra técnica de interconexión de redes es la conocida como peering, ya citada, en la cual existe un acuerdo de cooperación voluntaria entre una o más organizaciones. Esta técnica se caracteriza por no tener costo alguno de conexión, salvo el costo físico que implique la realización del cableado y/o necesidad de contar con interfaces de red en los routers. En este esquema, las organizaciones que intercambian altos volúmenes de tráfico, acuerdan una metodología de interconexión, de modo tal que el tráfico de red entre ambas Página 16 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ organizaciones, no tenga que ser transportado por sus proveedores de servicio de internet, sino gestionado directamente por las organizaciones. Para este tipo de conexiones generalmente se utilizan enlaces propios definidos entre las organizaciones, de modo tal de no tener que contratar el servicio a algún ISP. Una técnica muy buscada entre organizaciones, entidades y proveedores de servicio de internet para la interconexión es la utilización de puntos neutrales, también conocidos como Network Access Point (NAP) o Internet Exchange Point (IXP). Mediante esta técnica, diversas organizaciones acuerdan un punto común de interconexión de modo tal que permita intercambiar tráfico entre varios puntos a un costo inferior. Esta técnica puede brindar grandes beneficios a las organizaciones que generan tráfico hacia puntos interconectados a los NAP o IXP, dado que el tráfico no deberá pasar a través de diversos ISP para su transporte, sino que el mismo será canalizado localmente hacia el punto común de ruteo. Esta configuración generalmente acarrea costos de mantenimiento y administración, que son divididos entre todos los miembros del NAP con el fin de poder sustentar la arquitectura de comunicación. Para poder unirse a un NAP, generalmente se deben cumplir los requisitos de poseer un número de sistema autónomo propio y un propio bloque de direcciones IP, así como también se debe negociar con cada una de las organizaciones conectadas de qué modo intercambiarán datos. 2.4 Estructura de InterNet InterNet es una gran red de redes cuya arquitectura está basada en la interconexión de las mismas. Generalmente las conexiones forman una estructura jerárquica que va desde conexiones locales dentro de una misma ciudad, a conexiones nacionales e internacionales. Esta jerarquía de conexión permite estructurar a los ISP en niveles, de acuerdo a su grado y tipo de conectividad. Existen genéricamente 3 niveles de clasificación como se explica a continuación y se pueden observar en la figura 2.4.1: • Nivel/Tier 1: Las redes de los grandes operadores globales están clasificadas en esta categoría. Tienen la característica de poseer conexiones (generalmente de fibra óptica) por más de un continente. Desde este tipo de redes se puede acceder a cualquier red de InterNet, ya que es requisito que todas las redes de nivel 1 estén conectadas entre sí. Este tipo de redes forman el núcleo o backbone de InterNet y tienen la característica de no pagar a otro proveedor por servicios de transporte. • Nivel/Tier 2: Son las redes directamente conectadas a las de nivel 1, y que también poseen conexiones entre sí. Tienen un ámbito geográfico más acotado, generalmente a un mismo continente o un conjunto de países. Su principal función es proveer servicio a las redes de nivel 3. Página 17 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ • Nivel/Tier 3: Este tipo de redes tienen la característica de vender tráfico generalmente a organizaciones o usuarios residenciales. Son el último eslabón de la jerarquía y tienen un ámbito geográfico más acotado que las redes de nivel 2. Poseen conexiones entre ellas, a través de NAPs o IXPs y compran tráfico a redes de nivel 1 o 2. Figura 2.4.1: Estructura de la red InterNet en niveles Fuente: Curso de Ruteo avanzado dictado en Enero de 2014 por la Secretaría de Postgrado en la Facultad de Informática La comunicación entre las redes de distinto nivel, se realiza utilizando el protocolo BGP. Cada red posee su propio conjunto de redes IP, las cuáles están representadas por uno o más números de sistema autónomo. Adicionalmente existen políticas de tráfico que indican cómo deben canalizase los datos a través de cada vínculo. Página 18 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ 2.5 Sistemas Autónomos 2.5.1 Características Un sistema autónomo (Autonomous System o AS) está compuesto por un conjunto de redes bajo una política única y coherente de ruteo coordinada por un administrador. Cada sistema autónomo gestiona el modo en el que fluye le información desde su red hacia InterNet y viceversa. Cada Sistema autónomo está identificado por un número único que hace referencia a un conjunto de redes administradas. Este identificador referencia a dichas redes de manera única en la red InterNet. Hasta el año 2007, los números de sistema autónomo estaban definidos por un número entero de 16 bits, lo que permitía una asignación máxima de 65535 identificadores. Debido a la demanda, debió cambiarse el formato de los números con el fin de poder asignar una mayor cantidad de identificadores. La RFC 4983 define la implementación de números de sistema autónomo de 32 bits, los que se representan como un par de enteros x e y, números enteros de 16 bits. En la actualidad, los únicos números que se asignan son identificadores de 32 bits. 2.5.2 Clasificación Los Números de Sistema Autónomo son asignados por la InterNet Asigned Numbers Authority6 (IANA) a Registros regionales de InterNet (RIRs). Los RIR son los encargados de asignar regionalmente los números de sistema autónomo a las organizaciones finales. Una vez establecido, un sistema autónomo puede catalogarse de acuerdo a las siguientes definiciones, dependiendo de su modo de funcionamiento: • STUB: Representa a aquellos sistemas autónomos que solo se poseen un único camino o salida para su conexión a InterNet. Tiene la característica de ser siempre el último número en un camino de sistemas autónomos para representar una ruta. • Mutihomed Non-Transit: se encuentra conectado a través de varios caminos y poseen más de una conexión a InterNet. Característicamente, no rutea tráfico de terceros, es decir que todo el tráfico originado desde el propio sistema autónomo pertenece exclusivamente a sus redes. • Transit: Lleva tráfico de diferentes sistemas autónomos hacia InterNet. Esta clasificación corresponde generalmente a los ISP, los cuales lucran con dicho servicio. 6 La Internet Assigned Numbers Authority tiene autoridad sobre todo el espacio de direcciones IP utilizado en InterNet Página 19 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ La figura 2.5.2.1 muestra un ejemplo de cada una de las clasificaciones anteriores: Figura 2.5.2.1: Clasificación de sistemas autónomos 2.5.3 Requerimientos para obtener un bloque de Direcciones IP Con el fin de poder publicar un servicio en InterNet y que el mismo sea consumido por los usuarios, las organizaciones deben contar con un bloque de direcciones IP asignadas para tal fin. A través de dicho bloque de direcciones, serán identificados los servicios en la red InterNet. Generalmente las organizaciones solicitan a su ISP un subconjunto de direcciones asignadas al mismo durante el período de contrato que establezcan. Mediante dichas direcciones se publica el contenido en la red. El principal problema de este tipo de asignación, es que no permite el manejo de un esquema multiproveedor, ya que el rango de direcciones IP asignado temporalmente a la organización solo será ruteado globalmente mediante BGP a través de la infraestructura de comunicaciones de su ISP. Si la organización quisiera contratar el servicio de conectividad a un segundo proveedor, no podrá rutear el rango de direcciones que le hubiera asignado el primero, a través del nuevo ISP. Con el fin de subsanar este inconveniente, y en el caso de que la organización quisiera contar con un esquema multiproveedor, deberá solicitar y registrar un bloque de direcciones IPv4 propio, mediante la entidad de registro que lo alcance localmente de acuerdo a la distribución internacional establecida por IANA. Página 20 Universidad Nacional de La Plata – Facultad de Informática _____________________________________________________________________________________ El registro de direcciones IPv4 para la región de América Latina y Caribe está a cargo de LACNIC7. Cuando una organización desea obtener un bloque de direcciones IPv4, debe solicitarlo a dicha organización, cumpliendo como mínimo con los siguientes requisitos: • Proveer información detallada mostrando como el bloque solicitado será utilizado dentro de tres, seis y doce meses. • Entregar “planes de subneteo” por al menos un año, incluyendo máscaras de subred y números de hosts sobre cada subred. El uso de VLSM8 es requerido. • Entregar una descripción detallada de la topología de la red de la organización. • Realizar una descripción detallada de los planes de ruteo de la red, incluyendo los protocolos de ruteo a ser usados, así como también cualquier limitación existente. • En caso que el solicitante aún no cuente con un bloque IPv6[12] asignado por LACNIC, solicitarlo para cumplir con la política aplicable. Adicionalmente, la organización deberá demostrar que tiene planes de proveer servicios bajo un esquema multiproveedor, que utilizará y ocupará al menos el 50% de las direcciones IP asignadas en el plazo máximo de 1 año, y pagar el costo de registración inicial y renovación anual. A modo de referencia, de acuerdo al cuadro tarifario de LACNIC publicado en el siguiente sitio: http://www.lacnic.net/web/lacnic/tabla-de-precios, el costo a Noviembre/2015 asciende a U$D2500 (dos mil quinientos dólares) y U$D600 (seiscientos dólares) para registración inicial y renovación respectivamente.	1,819
W4361241643.txt_1	Open-Science-Pile	Open Science	Various open science	2,023	Supplementary Figure 1 from FOXQ1 Is Overexpressed in Colorectal Cancer and Enhances Tumorigenicity and Tumor Growth	None	English	Spoken	73	235	(Supplemental Figure 1) (A) (B) DLD-1 DLD 1 WiDr p21 40 100 Rel mRNA Re el mRNA 50 30 20 10 0 HEK293 800 p21 80 600 60 400 40 200 20 0 siRNA 0 siRNA siRNA p21 β-actin ((C)) HEK293 1000 H1299 120 100 p21 Rel m mRNA p21 β-actin CoLo320 p21 800 80 600 60 400 40 200 20 0 0 p21 80 p 21 β-actin p21 40 0 p21 β-actin.	8,606
https://openalex.org/W4366550170	OpenAlex	Open Science	CC-By	2,023	Functional and effective connectivity analysis of drug-resistant epilepsy: a resting-state fMRI analysis	Eric Jacob Bacon	English	Spoken	10,147	18,509	OPEN ACCESS OPEN ACCESS EDITED BY Ye Wu, Nanjing University of Science and Technology, China REVIEWED BY Elma M. Paredes-Aragon, National Institute of Neurology and Neurosurgery, Mexico Hanjiaerbieke Kukun, First Afﬁliated Hospital of Xinjiang Medical University, China CORRESPONDENCE Han Li leoincmu@gmail.com Shouliang Qi qisl@bmie.neu.edu.cn Eric Jacob Bacon1,2, Chaoyang Jin1, Dianning He1, Shuaishuai Hu3, Lanbo Wang4, Han Li3 and Shouliang Qi1,2* 1College of Medicine and Biological Information Engineering, Northeastern University, Shenyang, China, 2Key Laboratory of Intelligent Computing in Medical Image, Ministry of Education, Northeastern University, Shenyang, China, 3Department of Neurosurgery, Shengjing Hospital of China Medical University, Shenyang, China, 4Department of Radiology, Shengjing Hospital of China Medical University, Shenyang, China RECEIVED 10 February 2023 ACCEPTED 03 April 2023 PUBLISHED 20 April 2023 RECEIVED 10 February 2023 ACCEPTED 03 April 2023 PUBLISHED 20 April 2023 RECEIVED 10 February 2023 ACCEPTED 03 April 2023 PUBLISHED 20 April 2023 CITATION Bacon EJ, Jin C, He D, Hu S, Wang L, Li H and Qi S (2023) Functional and effective connectivity analysis of drug-resistant epilepsy: a resting-state fMRI analysis. Front. Neurosci. 17:1163111. doi: 10.3389/fnins.2023.1163111 Objective: Epilepsy is considered as a neural network disorder. Seizure activity in epilepsy may disturb brain networks and damage brain functions. We propose using resting-state functional magnetic resonance imaging (rs-fMRI) data to characterize connectivity patterns in drug-resistant epilepsy. Bacon EJ, Jin C, He D, Hu S, Wang L, Li H and Qi S (2023) Functional and effective connectivity analysis of drug-resistant epilepsy: a resting-state fMRI analysis. Front. Neurosci. 17:1163111. doi: 10.3389/fnins.2023.1163111 Methods: This study enrolled 47 participants, including 28 with drug-resistant epilepsy and 19 healthy controls. Functional and effective connectivity was employed to assess drug-resistant epilepsy patients within resting state networks. The resting state functional connectivity (FC) analysis was performed to assess connectivity between each patient and healthy controls within the default mode network (DMN) and the dorsal attention network (DAN). In addition, dynamic causal modeling was used to compute effective connectivity (EC). Finally, a statistical analysis was performed to evaluate our ﬁndings. The resting state functional connectivity (FC) analysis was performed to assess connectivity between each patient and healthy controls within the default mode network (DMN) and the dorsal attention network (DAN). In addition, dynamic causal modeling was used to compute effective connectivity (EC). Finally, a statistical analysis was performed to evaluate our ﬁndings. © 2023 Bacon, Jin, He, Hu, Wang, Li and Qi. TYPE Original Research PUBLISHED 20 April 2023 DOI 10.3389/fnins.2023.1163111 TYPE Original Research PUBLISHED 20 April 2023 DOI 10.3389/fnins.2023.1163111 TYPE Original Research PUBLISHED 20 April 2023 DOI 10.3389/fnins.2023.1163111 © 2023 Bacon, Jin, He, Hu, Wang, Li and Qi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Functional and effective connectivity analysis of drug-resistant epilepsy: a resting-state fMRI analysis OPEN ACCESS EDITED BY Ye Wu, Nanjing University of Science and Technology China REVIEWED BY Elma M. Paredes-Aragon, National Institute of Neurology and Neurosurgery, Mexico Hanjiaerbieke Kukun, First Afﬁliated Hospital of Xinjiang Medical University, China CORRESPONDENCE Han Li leoincmu@gmail.com Shouliang Qi qisl@bmie.neu.edu.cn Frontiers in Neuroscience 1. Introduction is acquired when the subject performs speciﬁc tasks. In contrast, a resting state is acquired when the subject rests (Logothetis, 2008). Understanding the properties of the brain network may help guide surgical intervention for better postoperative outcomes in drug-resistant epilepsy. A variety of resting state networks, each showing a deﬁnite spatial topography and putatively corresponding to a speciﬁc brain function (Yun et al., 2022). One of them, known as the default-mode network (DMN), is the most famous and essential network for the resting condition, as it consistently shows increased activity during rest than during active and passive cognitive tasks (Yun et al., 2022). The DMN areas typically comprise the posterior cingulate (PCC), medial prefrontal cortex (MPFC), and inferior parietal cortex (IPC) (Raichle et al., 2015). Same as DMN, the dorsal attention network (DAN) is another crucial resting-state fMRI network known to be active when performing speciﬁc tasks. However, DAN has also been proven to be associated with mesial temporal epilepsy in a resting state (Szczepanski et al., 2013). The DAN is centered on bilateral regions of the frontal and parietal cortex, including the frontal eye ﬁeld (FEF) and the intraparietal sulcus (IPS). Epilepsy is a common and serious neurological disorder that aﬀects around 50 million people worldwide (World Health Organization [WHO], 2023). Drug-resistant epilepsy (DRE) also known as refractory epilepsy is one of the most complex types of epilepsy. DRE is a complicated and heterogeneous condition in which seizures persist despite adequate trials of two or more carefully chosen and used antiepileptic drugs (AEDs). The term “adequate trial” refers to the use of AEDs at therapeutic doses for a suﬃcient period of time. The International League Against Epilepsy (ILAE) deﬁnes adequate trial failure as the persistence of disabling seizures despite the use of two tolerated, appropriately chosen and used AED schedules, either alone or in combination (Scheﬀer et al., 2017). Around 20%–30% of the epileptic population remains refractory to treatment and is considered to have drug-resistant epilepsy (Dalic and Cook, 2016). Diagnosis of DRE has remained challenging for neuroscientists and researchers. This diﬃculty can be explained by a variety of factors, including a lack of consensus deﬁnition, complex seizure semiology, limited diagnostic tools, comorbidities, and medication adherence issues (Jehi et al., 2022). For accurate diagnosis and management, a multidisciplinary approach and careful consideration of the patient’s individual circumstances are required. OPEN ACCESS This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Results: The FC analysis revealed signiﬁcant connectivity changes in patients giving 64.3% (18/28) and 78.6% (22/28) for DMN and DAN, respectively. Statistical analysis of FC was signiﬁcant between the medial prefrontal cortex, posterior cingulate cortex, and bilateral inferior parietal cortex for DMN. For DAN, it was signiﬁcant between the left and the right intraparietal sulcus and the frontal eye ﬁeld. For the DMN, the patient group showed signiﬁcant EC connectivity in the right inferior parietal cortex and the medial prefrontal cortex for the DMN. There was also bilateral connectivity between the medial prefrontal cortex and the posterior cingulate cortex, as well as between the left and right inferior parietal cortex. For DAN, patients showed signiﬁcant connectivity in the right frontal eye ﬁeld and the right intraparietal sulcus. Bilateral connectivity was also found between the left frontal eye ﬁeld and the left intraparietal sulcus, as well as between the right frontal eye ﬁeld and the right intraparietal sulcus. The statistical analysis of the EC revealed a signiﬁcant result in the medial prefrontal cortex and 01 Frontiers in Neuroscience frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 10.3389/fnins.2023.1163111 the right intraparietal cortex for the DMN. The DAN was found signiﬁcant in the left frontal eye ﬁeld, as well as the left and right intraparietal sulcus. Conclusion: Our results provide preliminary evidence to support that the combination of functional and effective connectivity analysis of rs-fMRI can aid in diagnosing epilepsy in the DMN and DAN networks. default mode network, dorsal attention network, resting-state functional magnetic resonance, functional connectivity, effective connectivity, epilepsy Frontiers in Neuroscience frontiersin.org 1. Introduction Treatment options that can help mitigate these challenges include optimizing AED therapy, surgical intervention, non-surgical interventions, complementary and alternative medicine, multidisciplinary care, and education and support. Recent technological advancements have simpliﬁed the process by providing numerous credible methods for analyzing and diagnosing brain diseases. Analyzing resting state fMRI (rs-fMRI) data remains challenging for neuroscientists. Brain connectivity modeling is a well-studied approach to illustrating brain function. One of the popular fMRI methods for studying brain networks is functional connectivity (rs-FC). Rs-FC measurements can detect coherent spontaneous neuronal activities within a brain network (van den Heuvel and HulshoﬀPol, 2010; Hlinka et al., 2011). This method has been explored by many researchers and is based on the temporal correlation between BOLD signals (Blood oxygenation level dependency) in distant brain regions. Lee et al. (2014) investigated changes in FC in brain networks for partial refractory epilepsy. The analysis was based on rs-fMRI using intrinsic connectivity contrast (ICC). Liu et al. (2021) also evaluated functional connectivity patterns in epilepsy associated with focal cortical dysplasia (FCD) to explore the underlying pathological mechanism of this disorder. Rs-FC is probably a good analytical approach to investigate rs-fMRI networks, but recent studies have highlighted some weaknesses and limitations of this approach (Buckner et al., 2013). These weaknesses could be covered by other analytical approaches, such as eﬀective connectivity (EC) analysis. This method provides adequate details on directed causal inﬂuences between diﬀerent regions of interest and covers some of the shortcomings observed during the FC analysis. Many studies have supported this using a similar technique to perform brain The development of new technology in neuroimaging has brought new insights into how brain disease can be diagnosed and cured. Many technologies for observing brain function non- invasively were developed and used to acquire brain signals. Among them, functional magnetic resonance imaging (fMRI) is considered one of the most prominent in the ﬁeld (Glover, 2011). This technology enabled the observation of regional brain activation by detecting the amount of oxygen in blood in each part of the brain. Here, we assume that the active region consumes more oxygen for energy. Resting state and task-based are two main kinds of fMRI utilized in the neuroimaging study. The main diﬀerence between the two lies in the acquisition procedure. Task-based fMRI 02 frontiersin.org Bacon et al. 1. Introduction 10.3389/fnins.2023.1163111 10.3389/fnins.2023.1163111 The others two samples were collected from an open-source website.1 The second sample corresponding to the second group of patients is provided by Thompson et al. (2020). Participants were patients with medically drug-resistant who had elected to undergo neurosurgical treatment for their epilepsy. network assessments and diagnose brain diseases. Using rs-fMRI, Wei et al. (2016) performed an eﬀective connectivity analysis to examine idiopathic generalized epilepsy-related changes in major neurocognitive brain networks. Jiang et al. (2018) also assessed the functional and eﬀective connectivity of the attention and default mode networks of rs-fMRI. Although, the EC method has many advantages and covers some limitations of FC analysis. However, several questions remain unanswered, so relatively few studies characterizing eﬀective or directed connectivity analysis of epileptic disease exist. From all these observations, concrete actions must be taken to create a more eﬃcient and precise method to eliminate these shortcomings. network assessments and diagnose brain diseases. Using rs-fMRI, Wei et al. (2016) performed an eﬀective connectivity analysis to examine idiopathic generalized epilepsy-related changes in major neurocognitive brain networks. Jiang et al. (2018) also assessed the functional and eﬀective connectivity of the attention and default mode networks of rs-fMRI. Although, the EC method has many advantages and covers some limitations of FC analysis. However, several questions remain unanswered, so relatively few studies characterizing eﬀective or directed connectivity analysis of epileptic disease exist. From all these observations, concrete actions must be taken to create a more eﬃcient and precise method to eliminate these shortcomings. The third sample is from the healthy control candidates collected by Gu et al. (2022). The subjects had no history of medical, neurological, or psychiatric disease. None of the subjects was taking medication at the time of testing. The image quality of all subjects and datasets was checked and controlled in a concise manner. The procedure and selection criteria were given in Figure 1. Our current study included a total of 47 subjects, including 28 patients and 19 healthy controls. All patients were collected and evaluated in accordance with standard principles. The evaluation included a detailed clinical history, a neurological examination, a complete neuropsychological assessment, a psychiatric assessment, and inter-ictal and ictal onset patterns in a long-term scalp video-electroencephalogram (video- EEG), MRI, and fMRI. They were all diagnosed with focal epilepsy, and the presurgical evaluation test results for each patient are shown in Table 1. 2.2. Data acquisition All rs-fMRI measurements were acquired and processed with a speciﬁc epilepsy protocol as used in the clinical routine. For sample 1, the MR images were acquired with a PET/MR scanner (SIGNA PET/MR; GE Healthcare, Waukesha, WI, USA) using a 16-channel head coil. The protocol included the following sequences: Sag 3D T1BRAVO (T1w; TR = 8.5 ms, TE = 3.3 ms, ﬂip angle = 12◦, voxel size = 0.469 × 0.469 × 1,000 mm3, FOV = 512 × 512). Resting-state BOLD images were acquired using a SIGNA PET/MR (TR = 2,000 ms, TE = 35 ms, Flip angle = 90 degrees, 3.5 × 3.4 × 4.0 mm3 voxel size). Sample 2 images were acquired as follows: T1-W structural scans were obtained on a 3T GE Discovery 750w (BRAVO, 32 ch head coil, TE = 3.376 ms, TR = 8.588 ms, Flip angle = 12 deg., 1.0 × 1.0 × 0.8 mm voxel size). Resting-state BOLD-fMRI sessions were obtained in a subset of subjects before implantation (4.8 min per session, 32 ch head coil, TR = 2,260 ms, TE = 30 ms, Flip angle = 80 degrees, 3.4 × 3.4 × 4.0 mm3 voxel size). 1. Introduction We propose to analyze functional and eﬀective connectivity based on rs-fMRI in this study. Our study aims to explore brain activities within the DMN and DAN of candidates with DRE. We hypothesize that studying functional and causal interactions within the brain network could help characterize and localize the epileptic zone. We also hypothesize that DRE candidates will have frequent seizures as a result of impaired brain communication caused by abnormal connectivity. Eﬀective and functional connectivity analysis will be performed to achieve our goal. The idea behind such an approach is to eﬀectively contribute to the diagnosis of the epileptogenic zone and facilitate surgical operations by making them more precise. This study was approved by the ethics committee of Shengjing Hospital of China Medical University and the informed consent was signed by the participant or a legal guardian/next of kin (for the participant under the age of 18). Our study is organized as follows. The functional connectivity analysis will ﬁrst be performed using the CONN toolbox to analyze rs-fMRI candidates. It will consist of performing a seed- based correlation analysis to assess the connectivity patterns between each patient and the healthy controls for DMN and DAN separately. Next, the eﬀective connectivity analysis will be performed using the DCM (Dynamic Causal Modeling) approach and the statistical Bayesian modeling inference, including Bayesian model selection and averaging. Finally, we evaluated our ﬁndings through statistical analysis. A two-sample t-test was conducted to diﬀerentiate connectivity within each network for the patients and healthy groups. This original study assesses the applicability of seed-based correlation and DCM analysis based on rs-fMRI data to diagnose epileptic networks. 1 openneuro.org Frontiers in Neuroscience 2.1. Participants The dataset in this study initially consisted of 71 subjects (38 patients and 33 HCs). To increase the population size in this study, the data was collected from three diﬀerent sources (2 samples of patients and a single sample of healthy controls). The ﬁrst sample consisted of 12 patients who underwent presurgical evaluation from January 2018 to July 2019 at Shengjing Hospital of China Medical University. The evaluation involved a detailed clinical history and neurological examination, complete neuropsychological evaluation, psychiatric assessment, inter-ictal and ictal onset patterns in long-term scalp video- electroencephalogram (video-EEG), magnetic resonance imaging (MRI), and fMRI results. All 12 patients underwent surgical resection for medically refractory epilepsy with histopathological conﬁrmation of FCD. Sample 3 data were collected on a 3 Tesla Prisma Siemens Fit scanner using a Siemens 20-channel receive-array coil. Anatomical images were acquired using an MPRAGE sequence (TR: 2,300 milliseconds, TE: 2.28 milliseconds, 1 mm isotropic spatial resolution, FOV: 256 millimeters, ﬂip angle: 8 degrees, matrix size: 256 × 256 × 192, acceleration factor: 2). Each scanning section consisted of an anatomical session, two 10-min resting- state sessions, and several 15-min sleep sessions. Blood oxygenation level-dependent (BOLD) fMRI data were acquired using an EPI sequence (TR: 2,100 milliseconds, TE: 25 milliseconds, slice 03 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 1 Criteria and the procedure for selecting subjects. Criteria and the procedure for selecting subjects. thickness: 4 mm, slices: 35, FOV: 240 mm, in-plane resolution: 3 mm × 3 mm). functional normalization, ART-based (Artifact Detection Tools) functional outlier detection and scrubbing, and functional smoothing (full-width-at-half maximum [FWHM 8-mm Gaussian kernel) were carried out in MNI-space. After pre-processing data, the processing step will follow. The CONN default processing was set as it implements the component-based noise correction method (CompCor) strategy for physiological and other noise source reduction, additional removal of movement, and temporal covariates, temporal ﬁltering, and windowing of the residual blood oxygen level-dependent (BOLD) contrast signal. During the processing step, resting-state signals will be extracted from the gray matter, and the cortex will be divided into diﬀerent regions of interest (ROI). Additionally, the mean time series of each ROI will be extracted as regressors, where other internal processing conditions will be speciﬁed along with the regressors. After that, individual connectivity maps will be created for each participant. Eight nodes derived from the networks were selected as the atlases or regions of interest. 2.1. Participants One can refer to Table 2 for the details concerning the networks and coordinate information of their nodes. 2 www.nitrc:projects/conn Frontiers in Neuroscience 2.5. Effective connectivity This study used the spectral dynamic causal modeling (DCM) approach to determine causal connectivity. All DCM analysis was performed with SPM12 according to the steps described in Sharaev et al. (2016). Spectral DCM analysis uses a neuronally plausible power-law model of the coupled dynamics of neuronal populations to generate complex cross spectra among the measured responses. Figure 1 shows the procedure included running general linear modeling followed by time series extraction, after which the DCM can be speciﬁed for each subject. The data used for the time series extraction includes data fully preprocessed during CONN analyses. After the GLM estimation, time series extraction was unsuccessful for 6 subjects, and we ﬁnally had 22 subjects for our further analysis. TABLE 1 Seizure onset zones for patients determined through the presurgical evaluation. TABLE 1 Seizure onset zones for patients determined through the presurgical evaluation. Patient Seizure onset zone 01 Left frontal 02 Left middle temporal 03 Left frontal 04 Left middle temporal 05 Left middle temporal 06 Left middle temporal 07 Left middle temporal 08 Left middle and lateral temporal 09 Right hippocampus and temporal lobe 10 Left temporal lobe 11 Left hippocampus and middle temporal 12 Right frontal 13 Left mesial temporal lobe 14 Right mesial temporal lobe 15 Right hippocampus 16 Left occipital lobe 17 Left frontal cystic mass 18 Left mesial temporal lobe 19 Left frontal encephalomalacia 20 Right mesial temporal lobe 21 Right anterior frontal lobe 22 Left mesial temporal lobe 23 Left mesial temporal lobe and frontal lobe 24 Left temporal pole 25 Right mesial temporal lobe 26 Right mesial temporal lobe and right frontal pole 27 Right mesial temporal lobe, Possible right frontal base 28 Left mesial temporal lobe The challenge of the eﬀective connectivity analysis was to assess the activities between diﬀerent brain regions for two rs-fMRI networks. To achieve this goal, neural modeling schemes must be speciﬁed, and this will help in making inferences as it provides details about the interactivities and the connectivity strength between diﬀerent regions (nodes) within a speciﬁc network TABLE 2 Network nodes and their coordinates. 2.3. Data processing The functional connectivity analysis in this study uses the CONN toolbox.2 CONN is a powerful and well-known neuroimaging toolbox that helps process task-related and rs- fMRI data (Whitﬁeld-Gabrieli and Nieto-Castanon, 2012; Schurz et al., 2014). For this study, we subdivided our method into three main steps: data preprocessing, processing, and statistical analysis (Figure 2). The classical CONN pre-processing procedure was performed using the default conﬁguration. It included the realignment and unwarping of the functional images, motion correction, slice-timing correction, and co- registration with the structural data (target resolution for functional images = 2 mm). Structural segmentation and normalization, 04 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 10.3389/fnins.2023.1163111 2.4. Functional connectivity control) and highlight the functional connectivity for the group- level analysis. For each region in DMN and DAN, a non-parametric test was used to compare the z-transformed connectivity maps of each drug-resistant patient and the healthy control group. This means that the comparison will be performed 224 (28 patients by 8 regions) times. The obtained results were considered signiﬁcant at a threshold of voxel-wise p < 0.001 uncorrected and cluster-level p < 0.05, false discovery rate (FDR) corrected for between-group comparisons. If one or more signiﬁcant clusters are found for each patient, we consider signiﬁcant connectivity changes. The functional connectivity analysis includes the ﬁrst-level and second-level analyses (Figure 2). The whole analysis was based on the seed-based connectivity (SBC) procedures deﬁned in CONN tool. In the ﬁrst-level analysis, for each subject, every region in DMN and DAN is used as a seed to conduct a seed-to-voxel functional connectivity analysis, such as to compute the correlation maps between the seed and voxels in the rest of the brain. DMN has 4 regions, including the posterior cingulate cortex (PCC), medial prefrontal cortex (MPFC), and bilateral inferior parietal cortex (LIPC and RIPC). DAN has four regions, including the frontal eye ﬁeld (FEF) and the intraparietal sulcus (IPS) at the left and right hemispheres. Finally, a z-transformed connectivity map is obtained for each region of each subject. Moreover, ROI-to-ROI connectivity analysis was performed to estimate the functional connectivity values between each pair of regions in DMN and DAN. The mean connectivity values of each region within each network were extracted for the patient and healthy control groups. In the second-level analysis, we ﬁrst set a contrast (a label of “1” is given to a drug-resistant patient, and “0” is given to a healthy Frontiers in Neuroscience Frontiers in Neuroscience 2.5. Effective connectivity Network Node Coordinate (x, y, z) Default mode network Medial prefrontal cortex (MPFC) 1, 55, −3 Left lateral inferior parietal (LIPC) −39, −77, 33 Right lateral inferior parietal (RIPC) 47, −67, 29 Posterior cingulate cortex (PCC) 1, −61, 38 Dorsal attention network Left frontal eye ﬁeld (L-FEF) −27, −9, 64 Right frontal eye ﬁeld (R-FEF) 30, −6, 64 Intraparietal sulcus (L-IPS) −39, −43, 52 Intraparietal sulcus (R-IPS) 39, −42, 54 TABLE 2 Network nodes and their coordinates. Frontiers in Neuroscience frontiersin.org 05 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 2 The ﬂowchart of the functional and effective connectivity study (FC: Functional connectivity, EC: Effective connectivity, and DCM: Dynamic causal modeling). FIGURE 2 The ﬂowchart of the functional and effective connectivity study (FC: Functional connectivity, EC: Effective connectivity, and DCM: Dynamic causal modeling). FIGURE 3 The speciﬁed models of effective connectivity in the default mode network (DMN) and dorsal attention network (DAN). FIGURE 2 The ﬂowchart of the functional and effective connectivity study (FC: Functional connectivity, EC: Effective connectivity, and DCM modeling). e functional and effective connectivity study (FC: Functional connectivity, EC: Effective connectivity, and DCM: Dynamic causal FIGURE 3 The speciﬁed models of effective connectivity in the default mode network (DMN) and dorsal attention network (DAN). FIGURE 3 The speciﬁed models of effective connectivity in the default mode network (DMN) and dorsal attention network (DAN). FIGURE 3 The speciﬁed models of effective connectivity in the default mode network (DMN) and dorsal attention network (DAN). to determine the best model that balances the data ﬁtting and model convolution (Rosa et al., 2010). Moreover, for the best model, Bayesian Model Averaging (BMA) was conducted (Hinne et al., 2020). The probability-weighted values obtained from the BMA (Friston et al., 2014; Bidhan and Mukesh, 2015). Therefore, six models were constructed for each subject. Figure 3 shows three models that were speciﬁed for the DMN and DAN. In addition, ﬁxed eﬀects (FFX) Bayesian Model Selection (BMS) was conducted (Friston et al., 2014; Bidhan and Mukesh, 2015). Therefore, six models were constructed for each subject. Figure 3 shows three models that were speciﬁed for the DMN and DAN. In addition, ﬁxed eﬀects (FFX) Bayesian Model Selection (BMS) was conducted 06 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 4 The clusters with different functional connectivity connected to the regions [as the seed) in the DMN and DAN network [here are the results of one patient (No. 2.5. Effective connectivity 02) as an example]. (A) Using PCC as the seed; (B) using MPFC as the seed; (C) using LIPC as the seed; (D) using RIPC as the seed; (E) using L-FEF as the seed; (F) using R-FEF as the seed; (G) using L-IPS as the seed; and (H) using R-IPS as the seed. FIGURE 4 FIGURE 4 The clusters with different functional connectivity connected to the regions [as the seed) in the DMN and DAN network [here are the results of one patient (No. 02) as an example]. (A) Using PCC as the seed; (B) using MPFC as the seed; (C) using LIPC as the seed; (D) using RIPC as the seed; (E) using L-FEF as the seed; (F) using R-FEF as the seed; (G) using L-IPS as the seed; and (H) using R-IPS as the seed. ABLE 3 An example of functional connectivity between one patient (No. 02) and healthy controls. TABLE 3 An example of functional connectivity between one patient (No. 02) and healthy controls. TABLE 3 An example of functional connectivity between one patient (No. 02) and healthy controls. Network Node (seed) Region with clusters Peak location (x, y, z) T-value The number of voxels DMN LIPC L-LOC −34 −78 36 7.96 681 RIPC R-LOC 48 −62 26 5.94 421 MPFC MedFC 0 58 −2 7.86 671 PCC Precu 6 −56 32 4.77 2,030 DAN L-FEF – – – – – – R-FEF R-PreCG 28 −6 34 5.04 54 L-IPS L-SPL −34 −48 44 7.13 912 R-IPS R-SPL 34 −44 52 6.72 805 L, left; R, right; IPC, lateral parietal; MPFC, medial prefrontal cortex; IPC, inferior parietal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, intraparietal sulcus; LOC, lateral occipital; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus. L, left; R, right; IPC, lateral parietal; MPFC, medial prefrontal cortex; IPC, inferior parietal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, occipital; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus. L, left; R, right; IPC, lateral parietal; MPFC, medial prefrontal cortex; IPC, inferior parietal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, intraparietal sulcus; LOC, lateral occipital; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus. showing abnormal clusters in patients. Frontiers in Neuroscience 2.5. Effective connectivity For DMN, signiﬁcant connectivity changes were observed in 64.3% (18/28) of patients. In comparison, 78.6% (22/28) of connectivity changes were observed for DAN. parameters models were quantitively analyzed using a classical one- sample t-test to examine the signiﬁcance of the non-zero values. 2.6. Statistical analysis Figure 4 and Table 3 illustrate an example of the observed changes within the DMN and DAN for a single patient. The abnormal locations included the lateral occipital, middle, and superior temporal gyrus, medial frontal cortex, and superior parietal lobule for DMN. Whereas, for DAN, these changes were seen in the precentral gyrus, superior parietal lobule, inferior temporal gyrus, inferior frontal gyrus, and temporal fusiform cortex. Comparing the individual performance of each network, the left lateral parietal lobule and posterior cingulate cortex nodes had the highest number of patients with signiﬁcant connectivity changes, with 42.4% (13/28) for DMN. The best connectivity performance for the DAN, on the other hand, was observed in the left intraparietal sulcus node, yielding 53.6% (15/28). Table 4 contains additional information about the ﬁndings of individual patient for both networks. After performing the functional and eﬀective connectivity analysis, a statistical test was performed to evaluate our study’s results. All the statistical tests were performed using SPSS. Once the average connectivity values among the pairs of each region were estimated in DMN and the DAN, these values were subjected to a two-sample t-test to verify the diﬀerence in connectivity between the patient group and healthy controls. frontiersin.org 3.1. Resting-state functional connectivity Selecting the DMN and DAN networks as regional seeds, the functional connectivity analysis revealed drug-resistant epilepsy Furthermore, the statistical analysis between the connectivity of patient and healthy control groups is shown in Figure 5. For 07 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 TABLE 4 The region with different functional connectivity to the regional seeds in DMN and DAN networks (obtained by comparing the generated functional connectivity mapping between each individual patient and the healthy control group). Sub. 3.1. Resting-state functional connectivity (-), no ﬁndings; L, left; R, right; LP, lateral parietal; MPFC, medial prefrontal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, intraparietal sulcus; OP, occipital lobe; LOC, lateral occipital; IFG, inferior frontal gyrus; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus; PostCG, post cingulate gyrus; ITG, inferior temporal gyrus; MTG, middle frontal gyrus; Cereb, cerebellum; Brain-S, brain stem; LG, lingual gyrus; AG, angular gyrus; oper, operculum; TFusC, temporal fusiform cortex. (-), no ﬁndings; L, left; R, right; LP, lateral parietal; MPFC, medial prefrontal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, intraparietal sulcus; OP, occipital lobe; LOC, lateral occipital; IFG, inferior frontal gyrus; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus; PostCG, post cingulate gyrus; ITG, inferior temporal gyrus; MTG, middle frontal gyrus; Cereb, cerebellum; Brain-S, brain stem; LG, lingual gyrus; AG, angular gyrus; oper, operculum; TFusC, temporal fusiform cortex. the DMN, the two-sample t-test was signiﬁcant between the medial prefrontal cortex and posterior cingulate cortex (p = 0.001) and the bilateral inferior parietal cortex (p = 0.0002 and 0.003). Signiﬁcant connectivity was also found between the posterior cingulate cortex and the right inferior parietal cortex (p = 1.4e-07). For DAN, the two-sample t-test was signiﬁcant between the left and right frontal eye ﬁelds (p = 1.2e-10). Additionally, signiﬁcance was found between the left and right intraparietal sulcus (p = 0.004). DMN and DAN, the fully connected models were the best for 4 of the 6 models speciﬁed for the patient and healthy control groups. For both networks, models 1 and 2 were the best for patients and healthy controls. At the group level, models 1 and 2 were the best for both networks in 22 out of 22 patients (90.9%). A similar scenario was observed for the healthy control group. Models 1 and 2 were the best for both networks in 17 out of 19 (89.5%) patients. For both networks, model 3 was weaker for the patient and healthy control groups. For the patient group, model 3 was better, with 3/22 (13.6%) and 4/22 (18.2%) for DMN and DAN, respectively. For the healthy control group, model 3 was the best for 2 out of 19 subjects (10.5%) for DMN and DAN. Frontiers in Neuroscience 3.1. Resting-state functional connectivity DMN DAN LIPC RIPC MPFC PCC L-FEF R-FEF L-IPS R-IPS – – – – – – – 02 L-LOC R-LOC MedFC Precu – R-PreCG L-SPL R-SPL 03 L-Caudate L- tri IFG – R-MTG L-FP R-ITG PC R-STG 04 L-LOC R-LOC MedFC Precu – R-PreCG L-SPL R-SPL 05 L-LG L-LG – L-LG L-LG L-LG L-LG L-LG 06 L-LOC R-LOC MedFC Precu – R-PreCG L-SPL R-SPL 07 - R-LOC MedFC Precu – R-PreCG L-SPL R-SPL 08 – – – – – – – – 09 L-oper IFG L-AG R-SPL L-oper IFG – – L-ITG R-SPL 10 R-LOC L-LOC R-LOC L-LOC – MedFC – – 11 R-OP L-ITG R-OP L-OP L-TFusC L-TFusC L-MFG L-FG oper 12 L-LOC L-LOC MedFC Precu – R-PreCG R-SPL R-SPL 13 L-LOC – – – – – R-ITG L-LOC 14 – – – – – – – R-SPL 15 – – – – – – R-SPL – 16 – – – – R-MTG L-OP L-PostCG – 17 – R-Cereb – – L-FP – – – 18 – – – – – – L-SPL – 19 – – R-Cereb – R-Cereb – L-SPL – 20 – – – – – – – – 21 – – FP l – L-PreCG – – – 22 R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb L-FP 23 – – R-Cereb – R-Cereb – – – 24 – – R-Cereb – R-SPL – – – 25 – – Brain-S Brain-S Brain-S Brain-S R-LOC Brain-S 26 R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb R-Cereb 27 – – – – L-PreCG – – – 28 R-SPL – R-PreCG - R-Cereb L-MTG L-MTG L-MTG (-), no ﬁndings; L, left; R, right; LP, lateral parietal; MPFC, medial prefrontal cortex; PCC, posterior cingulate cortex; FEF, frontal eye ﬁeld; IPS, intraparietal sulcus; OP, occipital lobe; LOC, lateral occipital; IFG, inferior frontal gyrus; MedFC, frontal medial cortex; Precu, precuneous cortex; FP, frontal pole; SPL, superior parietal lobule; PreCG, precentral gyrus; PostCG, post cingulate gyrus; ITG, inferior temporal gyrus; MTG, middle frontal gyrus; Cereb, cerebellum; Brain-S, brain stem; LG, lingual gyrus; AG, angular gyrus; oper, operculum; TFusC, temporal fusiform cortex. Sub. 3.2. Resting-state effective connectivity Bayesian model selection (BMS) for the patient and healthy control groups is shown in Figure 6. During BMS analysis, for the The results of BMA and the t-test are shown in Tables 5A, B. The one-sample t-test analysis found diﬀerent signiﬁcant 08 frontiersin.org frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 5 The strength of functional connectivity (mean and standard deviation) in the patient and healthy control groups and their comparisons. (A) The strength of FC for each group (patients and controls). (B) The p-value of FC comparisons between the patient and healthy control groups. p < 0.05, orange colors. FIGURE 5 The strength of functional connectivity (mean and standard deviation) in the patient and hea strength of FC for each group (patients and controls). (B) The p-value of FC comparisons be orange colors. FIGURE 5 The strength of functional connectivity (mean and standard deviation) in the patient and healthy control groups and their comparisons. (A) The strength of FC for each group (patients and controls). (B) The p-value of FC comparisons between the patient and healthy control groups. *p < 0.05, orange colors. connections for each network and group. For DMN, the patient group yielded signiﬁcant connectivity from the right inferior parietal cortex to the medial prefrontal cortex (EC = 0.02), bilateral connectivity was found between the medial prefrontal cortex and the posterior cingulate cortex (EC = 0.02), and bilateral connectivity in the inferior parietal cortex (EC = 0.02). Healthy controls did not show signiﬁcant connectivity. strength of interactions within the networks for each group is shown in Figure 7. Finally, the statistical analysis results of the eﬀective connectivity values between the patient and healthy control groups are shown in Tables 5C, D. For the DMN, the two-sample t-test was signiﬁcant from the middle prefrontal cortex to the left inferior parietal (p = 5.5E-05), and from the right inferior parietal cortex (p = 7.93E-09). For DAN, the patient group yielded signiﬁcant connectivity from the right frontal eye ﬁeld to the left intraparietal sulcus (EC = 0.2) and from the right intraparietal sulcus to the left frontal eye ﬁeld (EC = 0.03). Bilateral connectivity was found between the left frontal eye ﬁeld and the left intraparietal sulcus (EC = 0.04, EC = 0.2), as well as between the right frontal eye ﬁeld and the right intraparietal sulcus (EC = 0.13, EC = 0.03). 3.2. Resting-state effective connectivity For the DAN, the two-sample t-test was signiﬁcant from the left frontal eye ﬁeld to the intraparietal sulcus (p = 1.30E-10), the left intraparietal sulcus to the right frontal eye ﬁeld (p = 3.50E-07), and the right intraparietal sulcus to the left and left frontal eye ﬁelds (p = 2.20E-14 and 7.70E-11, respectively). For the healthy control group, signiﬁcant connectivity was observed from the left frontal eye ﬁeld to the left intraparietal sulcus (EC = 0.15), from the right frontal eye ﬁeld to the left intraparietal sulcus (EC = 0.02), and from the left intraparietal sulcus to the right frontal eye ﬁeld (EC = 0.04). Non-trivial connections have been considered, and self-connections in graphs have been ignored for simplicity. In addition, only signiﬁcant connections with a connection strength greater than 0.1 Hz and a probability greater than 0.95 were reported. The winning model that summarizes the Frontiers in Neuroscience Frontiers in Neuroscience frontiersin.org 4.1. Signiﬁcance and importance of this study Drug-resistant epilepsy remains one of the most severe cerebral diseases, and its diagnosis remains very tedious. Resting-state 09 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 6 The Bayesian model selection (BMS) at the single-group level. The columns represent the network (DAN and DMN), and each row represents the study group (patients and healthy controls). FIGURE 6 The Bayesian model selection (BMS) at the single-group level. The columns represent the network (DAN and DMN), and each row represents the study group (patients and healthy controls). et al., 2020). This assertion is conﬁrmed by Widjaja et al. (2013), who reported a decreased FC within the DMN in children with medically refractory epilepsy. Zhou et al. (2020) also highlighted the crucial role of the DAN network by investigating right temporal lobe epilepsy candidates. fMRI is an essential neuroimaging tool that has shown interesting results in diagnosing brain diseases. The purpose of this study is to investigate brain activities in rs-fMRI networks. The primary experiment of this study provides new information in the study of the connectivity of rs-fMRI networks of patients with drug- resistant epilepsy. Functional and eﬀective connectivity analysis approaches were used to assess connectivity behaviors in the default mode and the dorsal attention networks. Overall, the study provides satisfactory results. The DCM analysis performed well and showed connectivity for the DMN and DAN networks. The BMA values and the one-sample t-test analysis revealed signiﬁcant connectivity within patient and healthy control group networks. For DMN, the EC was signiﬁcant in the posterior cingulate cortex and medial prefrontal cortex for patients, while none was signiﬁcant for healthy controls. This result suggests that the posterior cingulate and the middle prefrontal cortex represent two regions with high sensitivity for drug-resistant candidates. In their study, Cook et al. (2019) conﬁrmed this observation in which the eﬀective connectivity of the DMN in The FC analysis showed abnormal activities in 18 out of 28 patients for the DMN compared with the DAN, which showed slightly higher performance showing activities in 22 out of 28 patients. This result supports that DMN and DAN are two crucial rs-fMRI networks to be evaluated during the presurgical analysis of candidates for refractory epilepsy (Blumenfeld et al., 2004; Hinne 10 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 10.3389/fnins.2023.1163111 These assessments have demonstrated noticeable progress. Boerwinkle et al. (2020) prospectively examined the inﬂuence of rs-fMRI on the organization of pediatric epilepsy surgery. Jiang et al. 4.3. Functional and effective connectivity performance comparison Brain connectivity is deﬁned as a pattern of interactions between the diﬀerent areas of the brain. Functional connectivity focuses on the temporal correlation among the activity of diﬀerent brain areas, while eﬀective connectivity relies on the causal interactions among the activity of diﬀerent brain areas. The fundamental diﬀerence between functional and eﬀective connectivity is the temporal implication of the source of the eﬀect, and this study has investigated the importance of functional and eﬀectivity analysis for the presurgical analysis of drug- resistant. We also tried to compare the best approach to be used when trying to investigate rs-fMRI patients. However, our evaluation revealed that both connectivity methods could answer our needs at diﬀerent levels. The descriptions above answer this study’s goal, combining both methods to bring out meaningful answers and facilitate surgical operations for drug- resistant candidates. Finally, for the DMN, the statistical analysis of the FC showed signiﬁcant diﬀerences between the medial prefrontal cortex and the posterior cingulate cortex and also between the posterior cingulate cortex and the right inferior parietal cortex. It was signiﬁcant for DAN between the left and right frontal eye ﬁeld and the intraparietal sulcus. Several researchers who used and compared both analyses approaches conﬁrmed this study’s assumption. Among them, Saetia et al. (2020) studied the interpretability of the eﬀective connectivity model compared with the functional connectivity model. Park et al. (2018) also evaluated the ensuing dynamic eﬀective connectivity in terms of the consistency of baseline connectivity within DMN using the rs-fMRI. They speculated that human brain networks at rest show dynamic functional connectivity induced by eﬀective dynamic connectivity, which can be modeled eﬃciently using dynamic causal modeling and hierarchical Bayesian inference. Mao et al. (2020) investigated the functional connectivity and eﬀective connectivity of the habenula in 34 subjects with irritable bowel syndrome (IBS) and 34 healthy controls and assessed the feasibility of diﬀerentiating IBS patients from healthy controls using a machine learning method. Moreover, for the DMN, the statistical analysis of the EC showed a signiﬁcant diﬀerence from the right inferior parietal cortex to the middle prefrontal and posterior cingulate cortex. Additionally, the diﬀerence was observed from the posterior cingulate cortex to the middle prefrontal cortex and the left inferior parietal cortex. This result suggests that the brain networks of the patient and control groups exhibit diﬀerent characteristics. Xiao et al. 4.3. Functional and effective connectivity performance comparison (2020) evaluated functional connectivity and topological properties of brain networks and found that their alterations were associated with neuropsychological disease. 4.1. Signiﬁcance and importance of this study (2018) also assessed the functional and causal connectivity of the attention networks and default mode network using rs-fMRI and revealed that epileptic activity might disrupt network interactions and further inﬂuence information communication. Zhang et al. (2011) also investigated epilepsy networks using resting-state fMRI, emphasized the importance of local network topology when investigating mechanisms underlying tumor-related epilepsy, and provided motivation for further investigation of the epilepsy process at the network level. patients with left temporal lobe epilepsy was assessed using the spectral DCM approach. Their study revealed connections between the posterior cingulate and medial prefrontal cortex (Cook et al., 2019). In addition, the patient group showed signiﬁcant connectivity from the right intraparietal sulcus and all other nodes for DAN. Left and right bilateral connectivity was also observed in the intraparietal sulcus. This result suggests that the right parietal lobule is a sensitive region in the DAN. Zhou et al. (2020) examined cognitive damage of the DAN in patients with right temporal lobe epilepsy (rTLE) and found a signiﬁcant diﬀerence in the right superior parietal lobule (SPL) and right precuneus (PCU) in patients with rTLE compared with healthy controls. Furthermore, comparing the results of the DCM analysis of the group of patients to that of healthy controls, a signiﬁcant diﬀerence in connectivity was also observed for the two networks. More activities were observed in the patient group compared with the control group. This hyper-activity observed within the group of patients may result from the frequent appearance of seizures in drug-resistant candidates. This last hypothesis (that abnormal interactions may cause frequent seizures in candidates for drug- resistant epilepsy) answers the objective of this study. Thus, the study provides evidence that eﬀective connectivity is a powerful presurgical and post-surgical analysis technique. This analytical approach was used by Jiang et al. (2018). In their study, the Granger causality eﬀective connectivity analysis approach was used to study the attention networks and default mode network of refractory participants. The speciﬁc disrupted networks appear to be associated with the speciﬁc cognitive characteristics of drug- resistant. Frontiers in Neuroscience 4.4. Limitations and challenges TABLE 5 The strength of effective connectivity (mean and standard deviation, in Hz) in the patient and healthy control groups and (A) Group Connection From MPFC From PCC From LIPC From RIPC Patient To MPFC 0 0.02 ± 0.001 0 0.2 ± 0.003 To PCC 0.02 ± 0.001 0 0.07 ± 0.005 0.13 ± 0.002 To LIPC 0.19 ± 0.004 0 0 0.02 ± 0.001 To RIPC 0 0 0.02 ± 0.001 0 Healthy control To MPFC 0 0 0 0.11 ± 0.001 To PCC 0 0 0.07 ± 0.001 0.11 ± 0.001 To LIPC 0.22 ± 0.003 0 0 0 To RIPC 0 0 0 0 (B) Group Connection From L-FEF From R-FEF From L-IPS From R-IPS Patient To L-FEF 0 0 0.2 ± 0.003 0.03 ± 0.001 To R-FEF 0 0 0.01 ± 0.002 0.01 ± 0.001 To L-IPS 0.04 ± 0.002 0.2 ± 0.004 0 0 To R-IPS 0.13 ± 0.006 0.13 ± 0.003 0 0 Healthy control To L-FEF 0 0 0.017 ± 0.001 0.15 ± 0.001 To R-FEF 0 0 0.04 ± 0.003 0.13 ± 0.002 To L-IPS 0.15 ± 0.002 0.02 ± 0.0 –0 0 To R-IPS −0.002 ± 0.001 −0.003 ± 0.0 0 0 (C) Connection From MPFC From PCC From LIPC From RIPC To MPFC 0 0.7 0 0.6 To PCC 0.7 0 0 0.07 To LIPC 5.50E-05 0 0 0.05 To RIPC 0 0 0.7 0 (D) Connection From L-FEF From R-FEF From L-IPS From R-IPS To L-FEF 0 0 0.9 2.20E-14 To R-FEF 0 0 3.50E-07 7.70E-11 To L-IPS 1.30E-10 0.8 0 0 To R-IPS 0.7 0.8 0 0 (A) Default mode network; (B) dorsal attention network; (C) the p-value of comparison of EC between the patient and healthy control groups in DMN; (D) the p-value of comparison of EC scale tests. Third, the lack of post-operative outcomes information in both patient samples makes comparison with our ﬁndings impossible. Fourth, diﬀerent acquisition procedures may have an adverse eﬀect on the interpretability of the result. The rs- fMRI population used in this study was acquired using slightly diﬀerential procedures. For the patients, sample 1 was obtained with the eyes closed, while sample 2 was obtained with the eyes open. For the healthy control group, a mixed eyes open-closed acquisition was used. 4.4. Limitations and challenges Our study proposes to assess the functional and eﬀective connectivity of the default mode and the dorsal attention networks, two networks known to present altered connectivity for epilepsy patients. The vital role of rs-fMRI in assessing altered brain connectivity for epilepsy patients has been investigated by several researchers. Despite the promising results obtained during our study, some challenges and limitations to this approach persist and must be overcome. First, a seed-based analysis is known as a relatively assumption-based approach. It requires the a priori selection of 11 frontiersin.org 10.3389/fnins.2023.1163111 Bacon et al. TABLE 5 The strength of effective connectivity (mean and standard deviation, in Hz) in the patient and healthy control groups and their comparisons. of effective connectivity (mean and standard deviation, in Hz) in the patient and healthy control groups and their comparisons. Frontiers in Neuroscience 4.4. Limitations and challenges This obvious acquisition diﬀerence may have an eﬀect on each candidate’s brain activity in sensorimotor and occipital regions (Wei et al., 2018). This limitation, however, a speciﬁc voxel, atlas, or network. However, the choice of seed may biologically bias the connectivity ﬁndings toward speciﬁc, smaller, or overlapping sub-systems rather than larger, distinct networks (Cole et al., 2010). The combination of a data and hypothesis-driven approach may provide a suitable answer to this problem (McKeown, 2000; Caulo et al., 2011). Second, the data size is crucial during analysis because a small sample size limits the statistical power. The sample size in this study is relatively small, which may impact the results (Grady et al., 2021). This issue must be considered preliminary and needs to be replicated in future studies with larger sample sizes and more detailed 12 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 FIGURE 7 The winning model at the group level. The number shows the connectivity parameters (Hz) of the winning model in the patients and healthy control groups represented by the columns. The rows represent the network type. The solid lines represent connectivity values greater than 0.1 Hz, and their thickness shows the size of the value. The dotted lines represent the connectivity values below 0.1 Hz. The orange represents the parameters with signiﬁcant non-zero values by a one-sample t-test (p < 0.05). The winning model at the group level. The number shows the connectivity parameters (Hz) of the winning model in the patients and healthy control groups represented by the columns. The rows represent the network type. The solid lines represent connectivity values greater than 0.1 Hz, and their thickness shows the size of the value. The dotted lines represent the connectivity values below 0.1 Hz. The orange represents the parameters with signiﬁcant non-zero values by a one-sample t-test (p < 0.05). has not resulted in statistical evidence and only a small portion of brain activity is aﬀected (Agcaoglu et al., 2019). This claim is supported by Patriat et al. (2013), who discovered that when the acquisition procedure was changed, only the visual network changed signiﬁcantly. abnormal functional connectivity for the DMN and DAN. In addition, dynamic causal modeling analysis has shown signiﬁcant eﬀective connectivity within both networks. Finally, the statistical analysis has demonstrated the connectivity diﬀerences within the networks of both patients and healthy control groups. 5. Conclusion Brain connectivity analysis has always been challenging for researchers. This study characterized drug-resistant epilepsy by assessing functional and eﬀective connectivity within resting state networks. The DMN and DAN networks were investigated at a subject and group level. Our analysis provided evidence of Data availability statement The datasets presented in this article are not readily available because of the requirement of the Ethics Committee of Shengjing Hospital of China Medical University (Shenyang, China). Requests to access the datasets should be directed to HL, leoincmu@gmail.com. 4.4. Limitations and challenges Our ﬁndings provide preliminary evidence to support that combining functional and eﬀective connectivity analysis may highly contribute to diagnosing altered brain networks in drug- resistant candidates. The results of this research may oﬀer new insight into the neuropathophysiological mechanisms of brain network dysfunction in drug-resistant epilepsy. In our subsequent studies, we will examine connectivity patterns between the DAN and DMN. Finally we also explored node-based connectivity analysis using the DCM method in this study. The expressiveness or complexity of the underlying neural model limits the interpretability of DCM. This complexity is constrained by the nature of the data at hand (Sadeghi et al., 2020). The strengths of the DCM approach lie in the hemodynamic model that links neuronal population ﬁring to BOLD data, which creates potential mismatches. 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Brain Mapp. 42, 204–219. doi: 10.1002/hbm.25217 Raichle, M. E., MacLeod, A. M., Snyder, A. Z., Powers, W. J., Gusnard, D. A., and Shulman, G. L. (2015). The brain’s default mode network. Annu. Rev. Neurosci. 38, 433–447. Ethics statement The studies involving human participants were reviewed and approved by the Ethics Committee of Shengjing Hospital of China 13 frontiersin.org Bacon et al. 10.3389/fnins.2023.1163111 Science Foundation of Liaoning Province (2020-BS-049 and 2021-YGJC-21), and the Fundamental Research Funds for the Central Universities (N2119010 and N2224001-10). Medical University. Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin. Funding This work was supported by the National Natural Science Foundation of China under Grant (82072008), the Natural Author contributions EB performed experiments and analyzed the data along with SQ and CJ. 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https://openalex.org/W3138224076	OpenAlex	Open Science	CC-By	2,021	Willingness to pay for cataract surgery and associated factors among cataract patients in Outreach Site, North West Ethiopia	Mohammed Assen Seid	English	Spoken	8,816	18,025	PLOS ONE PLOS ONE RESEARCH ARTICLE Mohammed Seid1, Amare Minyihun2, Gizachew Tilahun1, Asmamaw Atnafu2, Getasew AmareID2* 1 Department of Optometry, School of Medicine, College of Medicine and Health Science, University of Gondar, Gondar, Ethiopia, 2 Department of Health Systems and Policy, Institute of Public Health, College of Medicine and Health Science, University of Gondar, Gondar, Ethiopia a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 * getasewa23@gmail.com * getasewa23@gmail.com Methods Received: November 9, 2019 Accepted: March 2, 2021 Published: March 24, 2021 A cross-sectional outreach-based study was conducted on 827 cataract patients selected through a simple random sampling method in Tebebe Gion Specialized Hospital, North West Ethiopia, from 10/11/2018 to 14/11/2018. The data were collected using a contingent valuation elicitation approach to elicit the participants’ maximum willingness to pay through face to face questionnaire interviews. The descriptive data were organized and presented using summary statistics, frequency distribution tables, and figures accordingly. Factors assumed to be associate with a willingness to pay were identified using a Tobit regression model with a p-value of <0.05 and confidence interval (CI 6¼ 0). Peer Review History: PLOS recognizes the benefits of transparency in the peer review process; therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. The editorial history of this article is available here: https://doi.org/10.1371/journal.pone.0248618 OPEN ACCESS Citation: Seid M, Minyihun A, Tilahun G, Atnafu A, Amare G (2021) Willingness to pay for cataract surgery and associated factors among cataract patients in Outreach Site, North West Ethiopia. PLoS ONE 16(3): e0248618. https://doi.org/ 10.1371/journal.pone.0248618 OPEN ACCESS Citation: Seid M, Minyihun A, Tilahun G, Atnafu A, Amare G (2021) Willingness to pay for cataract surgery and associated factors among cataract patients in Outreach Site, North West Ethiopia. PLoS ONE 16(3): e0248618. https://doi.org/ 10.1371/journal.pone.0248618 OPEN ACCESS Citation: Seid M, Minyihun A, Tilahun G, Atnafu A, Amare G (2021) Willingness to pay for cataract surgery and associated factors among cataract patients in Outreach Site, North West Ethiopia. PLoS ONE 16(3): e0248618. https://doi.org/ 10.1371/journal.pone.0248618 Editor: Janhavi Ajit Vaingankar, Institute of Mental Health, SINGAPORE Introduction In Ethiopia, cataract surgery is mainly provided by donors free of charge through outreach programs. Assessing willingness to pay for patients for cataract surgery will help explain how the service is valued by the beneficiaries and design a domestic source of finance to sustain a program. Although knowledge concerning willingness to pay for cataract surgery is substantive for developing a cost-recovery model, the existed knowledge is limited and not well-addressed. Therefore, the study aimed to assess willingness to pay for cataract sur- gery and associated factors among cataract patients in Outreach Site, North West Ethiopia. Introduction Globally, cataract is the second leading cause of blindness, accounting for 33% (52.6 million) blind people next uncorrected refractive error [1, 2]. In Ethiopia, 50% of blindness is also caused by cataracts [3]. Cataract surgery is the only treatment modality [4, 5]. According to Vision 2020 initiatives, the targeted cataract surgical rate (CSR) varies globally from 2000– 5000 per million [6]. It was planned to be 2000 per million for African countries per year, but now it is less than 500 [7]. This underperformance was reasoned out by low patient demand, poor service delivery [7], low family income [8, 9], unable to afford surgical costs [8, 10, 11], logistical constraints, and fear of the surgery [9, 11], lack of knowledge, lack of family support [11], and time [9]. In 2005, the mean cost of the surgery per eye from various European coun- tries was US$ 843.5 [12]. By the year 2004, in the United States of America, the mean cost was estimated to be US$2525 [13]. Moreover, studies from Africa and South East Asia reported that the actual cost exceeds the community’s willingness to pay (WTP) for cataract surgery [14–18]. Overall, the proportion of WTP in Asia was very discrepant, which was ranged from 34% to 90%. Similarly, the mean amount of money participants willing to pay was varied from US$ 7 to US$ 968 [19–22]. However, the mean amount of money participants willing to pay in Africa was ranged from US$2.3 to US$18.5, which was relatively low as compared to Asian nations [19, 21]. In addition to regional variation, WTP for cataract surgery was higher among hospital-based studies and the urban population. However, the findings of outreach-based studies, which were conducted on the rural community, indicated that people are less willing to accept and pay for the surgery. Conclusively, the amount of money WTP is related to setting variation. Moreover, several studies showed that willingness to pay is affected by economic sta- tus, gender, age, family support, knowledge about the surgery, preoperative visual ability, fam- ily size, occupation, level of perceived need, and the characteristics of local eye care programs even if the majority of analysis model were fitted for the proportion of WTP rather than amount of money to pay [15, 17, 19–21, 23–26]. Besides, WTP is also health state dependant which is affected by the preference and perceived health status of participants. PLOS ONE PLOS ONE Factors affecting willingness to pay for cataract surgery Funding: The author(s) received no specific funding for this work. Conclusions Participants’ willingness to pay for cataract surgery in outreach Sites is much lower than the surgery’s actual cost. Early intervention and developing a cost-recovery model with multi- tiered packages attributed to the neediest people as in retired, less educated, severely dis- abled is strategic to increase the demand for service uptake and service accessibility. Competing interests: Authors have declared that no competing interests exist. Abbreviations: CV, Contingent Valuation; EDHS, Ethiopian Demographic and Health Survey; ETB, Ethiopian Birr; GDP, Gross Domestic Product; HEP, Health Extension Program; HEW, Health Extension Worker; HP, Health Post; HSDP, Health Sector Development Program; MWTP, Maximum Willingness to Pay. Results Copyright: © 2021 Seid et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The study involved 827 cataract patients, and their median age was 65years. About 55% of the participants were willing to pay for the surgery. The average amount of money willing to pay was 17.5USD (95% CI; 10.5, 35.00) and It was significantly associated with being still worker (β = 26.66, 95% CI: 13.03, 40.29), being educated (β = 29.16, 95% CI: 2.35, 55.97), free from ocular morbidity (β = 28.48, 95% CI: 1.08, 55.90), duration with the condition, (β = -1.69, 95% CI: -3.32, -0.07), admission laterality (β = 21.21, 95% CI: 3.65, 38.77) and remained visual ability (β = -0.29, 95% CI (-0.55, -0.04). Data Availability Statement: All relevant data are within the paper and its Supporting Information files. 1 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Study design and setting Outreach- based cross-sectional study was conducted. The campaign site was at Tebebe Gion Specialized Referral Hospital, Bahir Dar city, North West Ethiopia. There are only four Ter- tiary and general hospitals for about 14 million people in the catchment area, which provide comprehensive eye care services, including cataract surgical services in North West Ethiopia. However, all of them are urban-centred and not accessible for more than 80% of the commu- nity. Hence, the outreach program was planned for communities where cataract surgical ser- vices are not available and for peoples in disadvantaged areas with low socioeconomic status in North West Ethiopia. Initially, screening for cataracts was conducted in the nearby rural were- das, which are the third-level administrative units in the Ethiopian governance structure, before the campaign. During the screening, Cataract cases were diagnosed and informed about the condition and scheduled to attend the planned outreach program. As a result, a total of 1336 cataract cases were linked and admitted to the campaign from weredas of West Gojjam (628 patients), South Gondar (374 patients), Awi Zone (201 patients), Bahir Dar City (100 patients), and from East Gojjam (33 patients). Thus, more than 93% of cases were from rural weredas and district towns. Therefore, this study’s findings could apply to rural residents admitted for cataract surgery in outreach programs. Introduction This situation affects the research by over/under estimate result which may not reflect the actual price of the intervention [27–33]. In Ethiopia, it is known that the eye care service is urban-centred, which is not easily acces- sible for rural dwellers. As a result, cataract surgery is being delivered through outreach pro- grams, with the tremendous aid of donors, either free of charge or at a very low cost. Even if the program is essential and brought Socioeconomic welfare by reducing avoidable blindness associated with cataracts, this might not be financially sustainable for the long-run. Therefore, assessing WTP from the beneficiaries’ perspective is very important to understand how the users value the service and design a cost-recovery model that assures self-sustaining and high- volume surgical services [22]. Even if the knowledge regarding willingness to pay and the surgery’s actual cost is essential, the existing experience is not going through the intervention. Firstly, the study area’s existing 2 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE Factors affecting willingness to pay for cataract surgery knowledge is mainly centred on the proportion of willingness to pay rather than the amount of money [22]. Secondly, the amount of money WTP was not fully addressed with the appropri- ate analysis model. Thirdly, willingness to pay is highly setting-variant, which mainly varies with socioeconomic status and study setting. Consequently, it is difficult to apply the research finding done in other corners of the globe for this study area. Therefore, this study aimed to assess the willingness to pay for cataract surgery and associated factors among cataract patients at the Northwest Ethiopia outreach site. PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Study population and sampling technique All adult patients age 18 years admitted at the campaign Site, Tebebe Gion specialized Refer- ral Hospital, for cataract surgery was the study population except those who had communica- tion barriers (speech impairment and hearing impairment). The sample size was determined using single population mean estimation formula, by assuming a 95% confidence level, a standard deviation of WTP from the previous study con- ducted in Jimma, Southwest Ethiopia (US$7.5 (138.4 ETB)) (24), and a margin of error within 0.5$ around the parameter at 95% confidence level. The final computed sample size, including a 10% non-response rate, was 926. Thorough sampling procedures were followed to select the study participants. First, screen- ing for cataract was conducted in the nearby weredas so that 1336 cases were appointed for the surgery. The patients were documented and provided identification number. The appointment was based on the rate of surgical procedures planned per day, 170 cataract cases. Thus, the campaign lasted for eight consecutive days. Daily sample participants were calculated propor- tionally based on the daily patient flow. The sampling frame was designed based on the list of daily admitted patients. Hence, based on this proportion, the study participants were selected from cases prepared for admission using a simple random sampling method. 3 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE Factors affecting willingness to pay for cataract surgery Data collection procedures Data were collected using a structured questionnaire through face to face questionnaire inter- view from 10/11/2018 to 14/11/2018. The participants’ maximum willingness to pay (MWTP) for the cataract surgery interims of monetary value was estimated using a bidding game format of contingent valuation (CV) method [34–38]. The questionnaire was pretested on 47 patients at Ebnat primary hospital, the other equivalent outreach site. Measurements and variables Maximum willingness to pay (MWTP) for cataract surgery. Was the independent vari- able measured by questions prepared with a bidding game eliciting approach. After presenting the hypothetical scenario to assess how participants value the cataract surgery [39], the respon- dents were asked to accept or reject any positive price for cataract surgery. Those respondents who had accepted to pay some positive fees were asked their MWTP using the bidding format approach. The bidding game’s starting price was estimated from the actual cost of manual small incision cataract surgery per eye, 2824 ETB from a study done in Southern Ethiopia in 2010 [40]. In the Ethiopian context, the average household out pocket expenditure from the total health financing is around 35% [41, 42]. Based on this rate, the starting price we expected to be paid was 1000 Birr (0.35x2824birr). After presenting the case scenario, all participants were asked: PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 1. Yes Those who said no or unwilling to pay for the service were dropped for the next bidding game, which was used to assess the participants’ maximum willingness to pay for cataract sur- gery. But those who said yes for the first question were asked the maximum price they want to pay by the for the cataract surgery (Fig 1). Pre-operative visual ability/visual function. The visual system’s function can be measured objectively or subjectively. Visual acuity measurement is one of the objective indicators of the visual system functionality. However, it does not indicate the overall visual performance of an individual. Hence, we used the subjective measurement, the visual function/visual ability, to assess the overall visual performance. Hence it was measured by using adapted Visual Function 14 Index (VF-14) tool. Previously, numerous researchers used the tool as a standard. However, based on the socioeconomic and cultural perspective of their study population, the tool was adapted. The response option was prepared with 5-point Likert’s scale format. The scale ranged from the 0–4 value for the degree of visual difficulties’ unable to do’ (scale 0), ’great difficulty’ (scale1), ’moderate difficulty’ (scale 2), ’little difficulty’ (scale 3) and ’no difficulty’ (scale 4). Then each scale was multiplied by 25. So, the value of each response ranges from 0–100. After that, the factored amounts were summed up. Finally, VF index was computed by dividing the summed factored amounts to the number of checked boxes and the visual ability were categorized [43–45]. The calculation is elaborated across the studies, and the final classification is based on the International Council of Ophthalmology as normal or near-normal performance (50 visual ability score) and restricted performance (<50 visual ability score) [46]. Wealth index. Household wealth was measured by asking several questions on the posses- sion of different agricultural products, non-productive assets, and household facilities. The wealth index quintile was derived by Principal Component Analysis (PCA) [47, 48]. Data processing and analysis. The data were coded and entered into Epi Info version 7 and was exported to Stata Version 14 for analysis. The Tobit econometric model was used to PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 4 / 15 PLOS ONE Factors affecting willingness to pay for cataract surgery Fig 1. Iterative bidding technique to elicit willingness to pay for cataract surgery in outreach Sites, North West Ethiopia. 1. Yes First bidding questions: If the cost of cataract surgery for one eye is ETB 1000, are you willing to pay that amount? December 2018 Exchange rate: US$ 1 = ETB 27.8. h //d i /10 1371/j l 0248618 001 Fig 1. Iterative bidding technique to elicit willingness to pay for cataract surgery in outreach Sites, North West Ethiopia. First bidding questions: If the cost of cataract surgery for one eye is ETB 1000, are you willing to pay that amount? December 2018 Exchange rate: US$ 1 = ETB 27.8. https://doi.org/10.1371/journal.pone.0248618.g001 https://doi.org/10.1371/journal.pone.0248618.g001 analyze the determinants of willingness to pay and the maximum amount of money that patients willing to pay. This model is used to estimate parameters when the outcome variable is continuous in one end and has a constrained range in another arm [49–51]. The model is depicted as follow; y ¼ 1 0 ( PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Socio-demographic and economic characteristics of participants A total of 827 participants responded to the interviewer-administered questionnaire with a response rate of 89%. The median age of the participants was 65 years, with an interquartile range of 15. About 55% of the participants were age 65 years. The majority of the partici- pants, 498 (60.2%), were males. The significant portion of the participants, 675(82%), were from rural and had no formal education of 786(95%). Most of the participants were currently married 515(62.2%). About 84% of the participants had a family head role. About two-thirds of the participants, 565 (68.3%), were in a state of still working (Table 1). Participants’ willingness to pay for cataract surgery Fifty-five percent of the participants(456) were willing to pay something for the cataract sur- gery, and the average amount of money participants willing to pay was 500 ETB or 17.9 USD (10.9, 28.77). It was calculated as follows:- AMWTP ¼ X 827ðMWTP1 þ MWTP2þ þMWTP827Þ 827 ¼ 500 birr for cataract surgery service Note—1$USD = 27.9 birrs in May 2019 currency exchange, AMWTP is an average maxi- mum willingness to pay Health-related characteristics of participants The majority of the participants, 638 (77.1%), were not familiarized with cataract surgery either from themselves or their family members. Nearly 714(86%) of the participants had uni- lateral operable cataracts. Preoperatively, the participants’ mean visual ability score was 42.38, with a standard deviation of 23. The majority of the participants, 536(65%), had restricted visual performance, that was less than 50 visual ability score. The mean time spent with the condition was 2years. About 787(95%) of the participants did not report ocular co-morbidity other than cataract. Likewise, most of the participants, 720(87%), did not report any systemic illness. More than half of the participants, 466(56.3%), stated that they utilized community- based health insurance services (Table 2). Ethical considerations Ethical clearance was obtained from the ethical committee of the Institute of Public Health, College of Medicine and Health Science, and the University of Gondar. An official letter of permission was also obtained from Amhara Regional Health Bureau and Bahir Dar Tibebe Gion Specialized Referral Hospital. After explaining the purpose and importance of the study, informed consent was obtained from each study participant. Participants were also informed that they have the right to withdraw from the study if they face any inconvenience during the interview processes. i MWTP ¼ Bo þ B0 Xi þ e > 0 if MWTP 0 Y = Outcome variable; MWTP = Maximum WTP; Xi = Explanatory variables; βo = Slope; β’ = Coefficient; ε = error term; 1 = Success/Yes; 0 = Failure/No β’ = Coefficient; ε = error term; 1 = Success/Yes; 0 = Failure/No The model estimates the marginal effect of an explanatory variable on the expected value of the dependent variable. The descriptive data were organized and presented using summary statistics, frequency distribution tables, and figures accordingly. The Tobit model assumptions [52, 53] like normality, Multicollinearity (VIF = 1.23), and homoscedasticity (Breusch-pagan (hettest, X2 = 1.78, p = 0.18)) of error terms were checked. Regression coefficients (β), 95% confidence interval, and p-value of < 0.05 were used to measure a statistical association’s strength and presence. 5 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE Factors affecting willingness to pay for cataract surgery PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE PLOS ONE Factors affecting willingness to pay for cataract surgery Table 1. Socio-demographic characteristics of the study participants for WTP for cataract surgery in outreach Site, Northwest Ethiopia, 2019 (n = 827). Variables Willing to Pay Total (827) Yes (456) No (371) Frequency % Frequency % Frequency % Sex Male 292 35.3 206 24.9 498 60.2 Female 164 19.8 165 20 329 39.8 Age <65 years 220 26.6 154 18.6 374 45.2 65years 236 28.5 217 26.3 453 54.8 Residence Rural 373 45.1 302 36.5 675 81.6 Urban 83 10.1 69 8.3 152 18.4 Marital status Married 315 38.1 200 24.2 515 62.3 Widowed 111 13.4 118 14.3 229 27.7 Divorced 21 2.5 42 5.1 63 7.6 Single 9 1.1 11 1.3 20 2.4 Educational status Noformal education 430 52 356 43 786 95 Formal education 26 3.1 15 1.9 41 5 Working state Still working 349 42.2 216 26.1 565 68.3 Retired/unemployed 107 13 155 18.7 262 31.68 Household head Yes 387 46.8 308 37.2 695 84 No 69 8.4 63 7.6 132 16 Family size <5 305 36.9 281 34 586 70.86 5 151 18.3 90 10.8 241 29.14 Wealth status Poor 155 18.7 120 14.5 275 33.3 Medium 139 16.8 137 16.6 276 33.4 Rich 162 19.6 114 13.8 276 33.4 https://doi.org/10.1371/journal.pone.0248618.t001 Study participants whose one eye involved with cataracts were 36% (AOR = 0.64; 95% CI: 0.41, 0.98) less likely to pay some positive price for cataract surgery compared to those whose both eyes were involved. Besides, those who are not currently married were 1.55 (AOR = 2.55; 95% CI: 1.12, 2.14) times more likely to pay some positive cataract surgery price. The odds of participants who were still in working condition were 51% (AOR = 0.49; 95% CI: 0.35, 0.69) less likely to pay some positive price for cataract surgery than those who were not working (Table 3). Associated factors for willingness to pay some positive price for cataract surgery The study revealed that the eye involved with cataract, marital status, and occupation as signifi- cant variables for participants’ willingness to pay some positive price for cataract surgery. 6 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE Table 2. Health-related characteristics of the study participants for WTP for cataract surgery in outreach Site, North West Ethiopia, 2019 (n = 827). Variables Willing to pay Total Yes No Frequency % Frequency % Frequency % Familiarity with cataract surgery Yes 106 12.8 83 10.1 189 22.9 No 350 42.3 288 34.8 638 77.1 Eye involved with cataract One eye 408 49.3 306 37 714 86.3 Both eyes 48 5.8 65 7.9 113 13.7 Preoperative Visual ability <50 291 35.2 245 29.6 536 64.8 50 165 20 126 15.2 291 35.2 Time spent with condition <2years 147 17.8 96 11.6 243 29.4 2years 309 37.4 275 33.2 584 70.6 Self-reported ocular morbidity Yes 16 1.9 24 2.9 40 4.8 No 440 53.2 347 42 787 95.2 Self-reported Systemic illness Yes 55 6.7 52 6.3 107 12.9 No 401 48.4 319 38.6 720 87.1 Health insurance Yes 276 33.4 190 22.9 466 56.3 No 180 21.8 181 21.9 361 43.7 https://doi.org/10.1371/journal.pone.0248618.t002 retired/unemployed to active working state increases WTP for cataract surgery by 14.46 USD from mean willingness to pay (dy/dx = 14.46, 95% CI 7.48, 21.51). Participants who had formal education were WTP 29.16 USD more than participants who had no formal education (β = 29, 16, 95%CI 2.35, 55.97). retired/unemployed to active working state increases WTP for cataract surgery by 14.46 USD from mean willingness to pay (dy/dx = 14.46, 95% CI 7.48, 21.51). Participants who had formal education were WTP 29.16 USD more than participants who had no formal education (β = 29, 16, 95%CI 2.35, 55.97). retired/unemployed to active working state increases WTP for cataract surgery by 14.46 USD from mean willingness to pay (dy/dx = 14.46, 95% CI 7.48, 21.51). Participants who had formal education were WTP 29.16 USD more than participants who had no formal education (β = 29, 16, 95%CI 2.35, 55.97). Participants who had no ocular-comorbidity were willing to pay 28.48USD more than those with ocular co-morbidity, holding other variables constant (β = 28.48, 95%CI1.08, 55.90). The study depicted that being free from ocular morbidity increases WTP for cata- ract surgery by 14.26 USD from mean willingness to pay (dy/dx = 14.26, 95%CI 2.48, 26.04). Likewise, participants with unilateral cataracts were WTP 21.21USD more than those with bilateral cataracts, holding other variables constant (β = 21.21, 95%CI3.65, 38.77). PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Associated factors for willingness to pay for cataract surgery and their marginal effects The study showed that the still working participants were willing to pay 26.66USD more than the participants who retired or unemployed, keeping other variables constant (β = 26.66,95% CI13.03, 40.29). This variable’s marginal effect revealed that changing the working state from 7 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Factors affecting willingness to pay for cataract surgery Tobit/OLS/Truncated econometric analysis of factors associated with WTP for cataract surgery Tobit, OLS, and Truncated analysis were performed for factors related to WTP for cataract surgery. Educational status, working condition, visual ability, time spent with the disease, eye involved with cataract, and self-reported ocular co-morbidity were significant variables for WTP for cataract surgery by the Tobit model. In the OLS model, educational status, visual abil- ity, and cataract eyes were significantly associated with WTP for cataract surgery. Besides, par- ticipants’ educational level and visual ability were the significant variables for WTP (Table 5). PLOS ONE Table 3. Logistic regression for associated factors for willingness pay some positive price for cataract surgery in outreach Site, North West Ethiopia, 2019 (n = 827). Variable Category Willing to pay COR (95% CI) AOR (95% CI) P-value Yes No Age < 65 years 220 154 0.76 (0.58, 1.00) 0.86 (0.63, 1.19) 0.37 > = 65 years 236 217 1 Family head Yes 387 308 0.87 (0.60, 1.67) 0.70 (0.47, 1.06) 0.09 No 69 63 1 Residence Rural 373 302 1 Urban 83 69 1.03 (0.72, 1.46) 0.90 (0.59, 1.37) 0.63 Familiarity with cataract surgery Yes 106 83 1 No 350 288 1.05 (0.76, 1.46) 1.29 (0.71, 1.42) Self-reported ocular co-morbidity Yes 16 24 1 No 440 347 0.53 (0.28, 1.01) 0.54 (0.27, 1.06) 0.07 Eye involved with cataract One 408 306 0.55 (0.37,0.83) 0.64 (0.41, 0.98) 0.04 Both 48 65 1 Self-reported systemic co-morbidity Yes 55 52 1 No 401 319 0.84 9)0.56. 1.26 0.95 (0.61, 1.47) 0.82 Health Insurance Yes 276 190 0.68 (0.52, 0.90) 0.82 (0.61, 1.11) 0.20 No 180 181 1 Preoperative Visual Ability < 50 291 245 1 > = 50 165 126 0.91 (0.68, 1.21) 0.80 (0.59, 1.10) 0.17 Time spent with cataract < 2 years 147 96 0.73 (0.54, 0.99) 1.24 (0.90, 1.71) 0.19 > = 2 years 309 275 1 Wealth status Poor 155 120 1 Medium 139 137 1.27 (0.91, 1.78) 1.31 (0.91, 1.91) 0.15 Rich 162 114 0.91 (0.65, 1.27) 1.03 (0.72, 1.47) 0.87 Marital status Married 315 200 1 Not-married 141 171 1.91 (1.44, 2.54) 1.55 (1.12, 2.14) 0.008 Educational status No formal education 430 356 1 Formal education 26 15 0.70 (0.36, 1.34) 0.65 (0.32, 1.33) 0.24 Occupational status Retired/unemployed) 349 216 1 Still working 107 155 0.43 (0.32, 0.58) 0.49 (0.35, 0.69) 0.001 Family size < 5 305 281 1.55 (1.34, 2.10) 1.29 (0.92, 1.80) 0.14 > = 5 151 90 1 Constant 3.89 https //doi o g/10 1371/jo nal pone 0248618 t003 PLOS ONE This vari- able’s marginal effect showed that only having unilateral cataract increases WTP for cataract surgery by 11.20 USD from mean willingness to pay (dy/dx = 11.20, 95% CI 2.68, 19.73). It was also depicted that a one-year increment in years spent with the condition, the partici- pants’ WTP decreases by 1.69 USD, keeping the other variables constant (β = -1.69,95%CI -3.32, -0.07). This variable’s marginal effect indicated that increasing years lived with the con- dition by one year, it may decrease the WTP for cataract surgery by 1.12 USD from the mean (dy/dx = -1.12, 95%CI -2.05, -0.19). As the better eye’s visual ability increases by one unit, the participants’ WTP decreases by 0.29USD (β = -0.29, 95%CI -0.55, -0.04). This variable’s marginal effect revealed that increas- ing the visual ability in the better by one unit can decrease the WTP for cataract surgery by 0.16USD from mean willingness to pay (dy/dx = -0.16,95% CI-0.31, -0.02) (Table 4). 8 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Factors affecting willingness to pay for cataract surgery PLOS ONE Variables for MWTP Category β coefficients SE t-value p-value 95% CI Lower, upper Dy/dx(95% CI) Age N -0.34 0.26 -1.30 0.19 -0.85, 0.17 0.19(-0.48, .098) Being Married (ref, single) D 9.43 6.83 1.38 0.17 -3.98, 22.84 5.31(-2.14, 12.78) Being Urban (ref, rural) D -0.77 8.24 -0.09 0.93 -16.95, 15.42 2.94 (-5.80, 11.68) Formal education (ref, no formal education) D 29.16 13.66 2.14 0.03 2.35, 55.97 18.62(-0.22,37.46) Still working (ref, retired/unemployed) D 26.66 6.94 3.84 0.001 13.03, 40.29 14.49 (7.48, 21.51) Being Family head(ref, No) D 5.25 8.10 0.65 0.517 -10.65, 21.15 -0.44 (-9.61, 8.74) Visual ability N -0.29 0.13 -2.26 0.024 -0.55, -0.04 -0.16(0.31, -0.02) Time spent with the condition N -1.96 0.83 -2.37 0.018 -3.59, 0.34 -1.12 (-2.05, -0.19) Familiarity with cataract surgery (ref, yes) D 0.68 6.79 0.10 0.920 -12.67, 14.03 0.39 (-7.18, 7.96) No ocular co-morbidity (ref, yes) D 28.49 13.96 2.04 0.04 1.08, 55.90 14.26 (2.48, 26.04) No systemic illness (ref, yes) D 0.77 8.65 0.09 0.93 -16.2, 17.76 0.43 (-9.18, 10.06) Involved eye with cataract(ref, both eyes) D 21.21 8.95 2.37 0.018 3.65, 38.77 11.20 (2.68,19.73) Medium ref, poor) D -12.44 7.3 -1.70 0.089 -26.76, 1.89 -7.05(-15.16, 1.05) Rich (ref, poor) D -4.51 6.90 -0.65 0.51 -18.07, 9.04 -2.65 (-10.62, 5.31) Having Health insurance(ref, No) D 9.52 6.02 1.58 0.114 -2.28, 21.33 5.39 (-1.25, 12.04) Family size N 1.52 1.73 0.88 0.38 -1.87, 4.91 0.86 (-1.06, 2.79) Constant -29.02 28.07 -1.03 0.30 -84.12, 26.07 /Sigma 73.08 2.68 67.82 78.34 D, Dummy variable; N, numeric variable; ref, reference category; significant with p-value <0.05. https://doi.org/10.1371/journal.pone.0248618.t004 obit econometric analysis of factors associated with WTP for cataract surgery in outreach Site, North West Ethiopia,2019 aximum likelihood of Tobit econometric analysis of factors associated with WTP for cataract surgery in outreach Site, No volume surgical services. Although more than half of the patients admitted for cataract surgery (55%) were willing to pay something for the surgery, the average amount of money ready to pay for the surgery per eye was 500ETB (17.9USD). volume surgical services. Although more than half of the patients admitted for cataract surgery (55%) were willing to pay something for the surgery, the average amount of money ready to pay for the surgery per eye was 500ETB (17.9USD). Table 5. Tobit/OLS/Truncated econometric analysis of factors associated with WTP for cataract surgery in outreach Site, North West Ethiopia, 2019 (n = 827). PLOS ONE Variables Category Tobit Model OLS Model Truncated Model β coefficients SE P-value β coefficients SE P-value β coefficients SE P-value Age N -0.34 0.26 0.19 -0.23 0.15 0.13 -0.45 0.30 0.13 Being Married (ref, single) D 9.43 6.83 0.17 2.57 4.01 0.56 -11.9 7.72 0.12 Being Urban (ref, rural) D -0.77 8.24 0.93 -1.8 4.86 0.71 -9.1 9.17 0.32 Formal education (ref, no formal education) D 29.16 13.66 0.03 22.7 8.28 0.006 30.3 13.49 0.03 Still working (ref, retired/unemployed) D 26.66 6.94 0.001 12.5 4.02 0.002 -0.46 8.20 0.96 Being Family head(ref, No) D 5.25 8.10 0.517 0.36 4.78 0.94 -11.02 8.92 0.22 Visual ability N -0.29 0.13 0.024 -0.79 0.08 0.008 -0.35 0.14 0.01 Time spent with the condition N -1.96 0.83 0.018 -0.79 0.46 0.09 1.1 0.96 0.29 Familiarity with cataract surgery (ref, yes) D 0.68 6.79 0.920 -0.46 4.05 0.91 -1.8 7.53 0.82 No ocular co-morbidity (ref, yes) D 28.49 13.96 0.04 13.4 7.81 0.09 11.4 17.53 0.52 No systemic illness (ref, yes) D 0.77 8.65 0.93 0.25 5.10 0.96 -2.3 9.88 0.81 Involved eye with cataract(ref, both eyes) D 21.21 8.95 0.018 11.9 5.14 0.02 12.7 10.86 0.24 Medium ref poor) D -12 44 7 3 0 089 -6 9 4 33 0 11 -0 46 8 09 0 52 volume surgical services. Although more than half of the patients admitted for cataract surgery (55%) were willing to pay something for the surgery, the average amount of money ready to pay for the surgery per eye was 500ETB (17.9USD). Table 4. Maximum likelihood of Tobit econometric analysis of factors associated with WTP for cataract surgery in outreach Site, North West Ethiopia,2019 (n = 827). Discussion This study aimed to assess WTP for cataract surgery and associated factors to provide substan- tive evidence for developing a cost-recovery model that assures self-sustaining and high- 9 / 15 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Factors affecting willingness to pay for cataract surgery PLOS ONE PLOS ONE volume surgical services. Although more than half of the patients admitted for cataract surgery (55%) were willing to pay something for the surgery, the average amount of money ready to pay for the surgery per eye was 500ETB (17.9USD). Table 4. Maximum likelihood of Tobit econometric analysis of factors associated with WTP for cataract surgery in outreach Site, North West Ethiopia,2019 (n = 827). Variables for MWTP Category β coefficients SE t-value p-value 95% CI Lower, upper Dy/dx(95% CI) Age N -0.34 0.26 -1.30 0.19 -0.85, 0.17 0.19(-0.48, .098) Being Married (ref, single) D 9.43 6.83 1.38 0.17 -3.98, 22.84 5.31(-2.14, 12.78) Being Urban (ref, rural) D -0.77 8.24 -0.09 0.93 -16.95, 15.42 2.94 (-5.80, 11.68) Formal education (ref, no formal education) D 29.16 13.66 2.14 0.03 2.35, 55.97 18.62(-0.22,37.46) Still working (ref, retired/unemployed) D 26.66 6.94 3.84 0.001 13.03, 40.29 14.49 (7.48, 21.51) Being Family head(ref, No) D 5.25 8.10 0.65 0.517 -10.65, 21.15 -0.44 (-9.61, 8.74) Visual ability N -0.29 0.13 -2.26 0.024 -0.55, -0.04 -0.16(0.31, -0.02) Time spent with the condition N -1.96 0.83 -2.37 0.018 -3.59, 0.34 -1.12 (-2.05, -0.19) Familiarity with cataract surgery (ref, yes) D 0.68 6.79 0.10 0.920 -12.67, 14.03 0.39 (-7.18, 7.96) No ocular co-morbidity (ref, yes) D 28.49 13.96 2.04 0.04 1.08, 55.90 14.26 (2.48, 26.04) No systemic illness (ref, yes) D 0.77 8.65 0.09 0.93 -16.2, 17.76 0.43 (-9.18, 10.06) Involved eye with cataract(ref, both eyes) D 21.21 8.95 2.37 0.018 3.65, 38.77 11.20 (2.68,19.73) Medium ref, poor) D -12.44 7.3 -1.70 0.089 -26.76, 1.89 -7.05(-15.16, 1.05) Rich (ref, poor) D -4.51 6.90 -0.65 0.51 -18.07, 9.04 -2.65 (-10.62, 5.31) Having Health insurance(ref, No) D 9.52 6.02 1.58 0.114 -2.28, 21.33 5.39 (-1.25, 12.04) Family size N 1.52 1.73 0.88 0.38 -1.87, 4.91 0.86 (-1.06, 2.79) Constant -29.02 28.07 -1.03 0.30 -84.12, 26.07 /Sigma 73.08 2.68 67.82 78.34 D, Dummy variable; N, numeric variable; ref, reference category; significant with p-value <0.05. https://doi.org/10.1371/journal.pone.0248618.t004 Table 5. Tobit/OLS/Truncated econometric analysis of factors associated with WTP for cataract surgery in outreach Site, North West Ethiopia, 2019 (n = 827). PLOS ONE Variables Category Tobit Model OLS Model Truncated Model β coefficients SE P-value β coefficients SE P-value β coefficients SE P-value Age N -0.34 0.26 0.19 -0.23 0.15 0.13 -0.45 0.30 0.13 Being Married (ref, single) D 9.43 6.83 0.17 2.57 4.01 0.56 -11.9 7.72 0.12 Being Urban (ref, rural) D -0.77 8.24 0.93 -1.8 4.86 0.71 -9.1 9.17 0.32 Formal education (ref, no formal education) D 29.16 13.66 0.03 22.7 8.28 0.006 30.3 13.49 0.03 Still working (ref, retired/unemployed) D 26.66 6.94 0.001 12.5 4.02 0.002 -0.46 8.20 0.96 Being Family head(ref, No) D 5.25 8.10 0.517 0.36 4.78 0.94 -11.02 8.92 0.22 Visual ability N -0.29 0.13 0.024 -0.79 0.08 0.008 -0.35 0.14 0.01 Time spent with the condition N -1.96 0.83 0.018 -0.79 0.46 0.09 1.1 0.96 0.29 Familiarity with cataract surgery (ref, yes) D 0.68 6.79 0.920 -0.46 4.05 0.91 -1.8 7.53 0.82 No ocular co-morbidity (ref, yes) D 28.49 13.96 0.04 13.4 7.81 0.09 11.4 17.53 0.52 No systemic illness (ref, yes) D 0.77 8.65 0.93 0.25 5.10 0.96 -2.3 9.88 0.81 Involved eye with cataract(ref, both eyes) D 21.21 8.95 0.018 11.9 5.14 0.02 12.7 10.86 0.24 Medium ref, poor) D -12.44 7.3 0.089 -6.9 4.33 0.11 -0.46 8.09 0.52 Rich (ref, poor) D -4.51 6.90 0.51 -3.6 4.15 0.38 -6.1 7.60 0.42 Having Health insurance(ref, No) D 9.52 6.02 0.114 6.7 3.55 0.06 8.2 6.79 0.23 Family size N 1.52 1.73 0.38 0.92 1.03 0.38 0.10 1.89 0.96 Constant -29.02 28.4 94.2 31.3 https://doi org/10 1371/journal pone 0248618 t005 actors associated with WTP for cataract surgery in outreach Site, North West Ethiopia, 2019 (n = 827). PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 10 / 15 PLOS ONE Factors affecting willingness to pay for cataract surgery This result was comparable with outreach based studies done in Jimma, Ethiopia (US$12.4 [22], and in Northern Nigeria (US$18.5) [19]. On the other hand, it is greater than the cam- paign based studies conducted in Tanzania (US$2.3 [20], Malawi (3US$), [21], and Nepal (US $7) [17]. Despite design similarity, they were conducted on a minimal sample size selected with non-probability sampling. This may be a reason for the discrepancy. However, this result was lower than the studies done in Hong Kong (US$ 552) [23] and China (USD 968) [25]. PLOS ONE Both studies involved cataract patients on the waiting list in hospitals and used a bidding for- mat for eliciting the willingness to pay. Relatively, their target price was higher as compared to the present study. Having a difference in target price and research setting might be taken as a reason for the variation. Overall, this amount of money willing to pay matches the subsidized price for cataract sur- gery at General and Tertiary Hospitals. However, it was significantly lower than the current cost of the surgery area, 2824 ETB(101.56USD) [40], and the proposed target price by 50%. Lack of money, lack of knowledge to access the services, and waiting for the campaign were the main reason cited to be unwilling to pay and unable to seek eye care services previously. Currently, cataract surgery is being delivered for districts through outreach programs, with the tremendous aid of donors, free of charge. Nevertheless, this might not be financially sus- tainable for the long-run. This study’s findings imply that if the government or private sectors plan to sustain the program with domestic resources, it is strategic to develop an inclusive cost-recovery model that can assure services accessibility, self-reliance, and equity. The Ara- vind Eye Care System in India is the best lesson: providing high-quality and high-volume (350,000 eye operations per year) services to those who can afford to pay market rates and then uses the profits to fund care for those who cannot. Patients who cannot afford to pay are given cataract surgery for free. However, the government reimburses Aravind US$10 for each procedure [54]. The Ethiopian health care financing policy considers the systematizing fee waiver system, while such approaches are not applied for cataract surgical services so far. Moreover, the WTP’s comparability with the current price of cataract surgery set for Gen- eral and Tertiary Hospitals indicates that there is a probability of integrating the service in the existing health care system at the district level if the Ethiopian government invests on human resource development and infrastructure. The working state’s impact on willingness to pay is depicted that the participants who were still working were WTP 26.66USD more than the participants who were retired/unemployed. This result is supported by a study conducted in rural china [15] though some studies con- cluded that willingness to pay was not affected by the working state [18, 21, 25]. PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 PLOS ONE This relation- ship can be explained in a way that employees can be related to greater productivity. This implies that retired/ unemployed persons would be a potential target for cost- recovery. Persons with formal education were 29.16 USD more WTP than those who had no formal education for surgery. This finding is consistent with a study done in China [24]. This may suggest that educated persons may value their vision more and may have good knowledge about the service’s cost-benefit aspects. However, many studies found that WTP for cataract surgery was not significantly affected by literacy because these studies were done in nations where there is higher educational coverage [15, 18, 22, 23]. Hence, significant variation based on education might not be observed. This result indicated that extra efforts in outreach pro- grams need to be invested in prospering illiterate persons’ knowledge with non-printed media. Patients with no ocular-comorbidity and unilateral cataracts were more willing to pay than those with ocular co-morbidity and bilateral cataract. Previous studies also reported that pre- existing eye diseases and bilateral cataracts affect willingness to pay for cataract surgery [25]. This may be explained so that persons with co-morbidity and bilateral cataract might have poor visual performance. Hence, their productivity and economy may be compromised, PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 11 / 15 PLOS ONE Factors affecting willingness to pay for cataract surgery making them desperate to have good vision and uptake the services. On the other hand, as the visual performance of a better eye decreases by one unit, the participants’ WTP increases by 0.29USD. Similarly, a study reported that persons with poor visual performance were more willing to pay for the surgery [23]. The justification might be that patients with poor visual ability can be highly ambitioned to enhance their daily base activities regardless of their ability to pay. On the contrary, two studies found that as the preoperative visual acuity becomes more impaired, the willingness to pay decreases. The visual functionality was measured objectively [21, 24]. These findings imply that cataract surgery’s subsidization is substantive for patients with a severe visual disability, ocular-comorbidity preexisted condition, and bilateral cataract. Thus, even if resource utilization efficiency is sacrificed, equity among services consumers might be logical. The study also depicted that one year increment in years lived with visual impairment, the participants WTP decreases by 1.69 USD. PLOS ONE The explanation might be that as the duration increase without intervention, the severity of visual disability and complication associated with cataract can be increased. This increases the psycho-socio-economic burden of the condition. Hence, it ends up with a poor vision-related quality of life and poor ability to pay. This implies that early intervention may promote a greater willingness to pay and service utilization. Limitation of the study The method used only shows the service’s values or benefits, which didn’t consider the full eco- nomic evaluation approach. The other limitation of this study is the strategic response bias in which the respondents will intentionally overestimate or underestimate the price, which is not their actual price. We tried to minimize this bias using a well-stated case scenario and Bidding game format as an elicitation approach. PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 Conclusion Willingness to pay for cataract surgery among adults with operable cataracts in outreach sites is much lower than the surgery’s actual cost. People with a state of having a formal education, working state, less time spent with the condition, free from co-morbidity, and bilateral cata- racts were more willing to pay for cataract surgery. Therefore, the provision of cataract surgical services by offering a multi-tiered package attributed to the population’s characteristics is essential and applicable. Acknowledgments The authors would like to thank all respondents for their willingness to participate in the study. We are also grateful to the Amhara public health institute, Tebebe Gion Specialized PLOS ONE \| https://doi.org/10.1371/journal.pone.0248618 March 24, 2021 12 / 15 PLOS ONE Factors affecting willingness to pay for cataract surgery Referral Hospital, and the University of Gondar for material support. Finally, our appreciation goes to data collectors for their unreserved contribution in data collection activities. Author Contributions Conceptualization: Mohammed Seid. Data curation: Mohammed Seid. Formal analysis: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Methodology: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Software: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Writing – original draft: Mohammed Seid. Writing – review & editing: Amare Minyihun, Asmamaw Atnafu, Getasew Amare. Conceptualization: Mohammed Seid. Data curation: Mohammed Seid. Data curation: Mohammed Seid. Formal analysis: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Formal analysis: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Methodology: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Software: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Software: Mohammed Seid, Amare Minyihun, Gizachew Tilahun, Asmamaw Atnafu, Getasew Amare. Writing – original draft: Mohammed Seid. Writing – original draft: Mohammed Seid. Writing – original draft: Mohammed Seid. Writing – review & editing: Amare Minyihun, Asmamaw Atnafu, Getasew Amare. 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https://openalex.org/W2938945084	OpenAlex	Open Science	CC-By	2,019	Correction: Effect of maturity and mineralogy on fluid-rock reactions in the Marcellus Shale	John Pilewski	English	Spoken	812	1,500	Correction: Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale John Pilewski,a Shikha Sharma,a Vikas Agrawal,a J. Alexandra Hakalab and Mengling Y. Stuckmanb Correction for ‘Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale’ by Joh Pilewski et al., Environ. Sci.: Processes Impacts, 2019, DOI: 10.1039/c8em00452h. In the original article, the %R0 values on the x-axis of Fig. 3C were incorrect. The Figure with the corrected values is shown below The authors apologise for this error and state that this error does not aﬀect any of the scientic ndings and interpretations. The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers. Fig. 3 (C) Plot of the relationship between total target VOCs, total target organic acids, TOC, and %R0. The lines indicate the direction of the dat trend and do not represent a calculated ﬁt of the data. Cite this: Environ. Sci.: Processes Impacts, 2019, 21, 905 DOI: 10.1039/c9em90015b rsc.li/espi Environmental Science Processes & Impacts CORRECTION Open Access Article. Published on 10 April 2019. Downloaded on 10/24/2024 11:52:23 AM. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. View Article Online View Journal \| View Issue Correction: Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale John Pilewski,a Shikha Sharma,a Vikas Agrawal,a J. Alexandra Hakalab and Mengling Y. Stuckmanb Correction for ‘Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale’ by Joh Pilewski et al., Environ. Sci.: Processes Impacts, 2019, DOI: 10.1039/c8em00452h. In the original article, the %R0 values on the x-axis of Fig. 3C were incorrect. The Figure with the corrected values is shown below The authors apologise for this error and state that this error does not aﬀect any of the scientic ndings and interpretations. The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers. Fig. 3 (C) Plot of the relationship between total target VOCs, total target organic acids, TOC, and %R0. The lines indicate the direction of the dat trend and do not represent a calculated ﬁt of the data. Cite this: Environ. Sci.: Processes Impacts, 2019, 21, 905 DOI: 10.1039/c9em90015b rsc.li/espi Environmental Science Processes & Impacts CORRECTION Open Access Article. Published on 10 April 2019. Downloaded on 10/24/2024 11:52:23 AM. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. View Article Online View Journal \| View Issue aWest Virginia University Department of Geology & Geography, 330 Brooks Hall, 98 Beechurst Ave., Morgantown, WV 26506, USA. E-mail: shikha.sharma@mail.wvu.edu; Tel: +1-304-293-6717 bNational Energy Technology Laboratory, U. S. Department of Energy, Pittsburgh, PA 15236, USA This journal is © The Royal Society of Chemistry 2019 CORRECTION Correction: Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale Correction: Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale Cite this: Environ. Sci.: Processes Impacts, 2019, 21, 905 John Pilewski,a Shikha Sharma,a Vikas Agrawal,a J. Alexandra Hakalab and Mengling Y. Stuckmanb Correction for ‘Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale’ by John Pilewski et al., Environ. Sci.: Processes Impacts, 2019, DOI: 10.1039/c8em00452h. In the original article, the %R0 values on the x-axis of Fig. 3C were incorrect. The Figure with the corrected values is shown below. The authors apologise for this error and state that this error does not aﬀect any of the scientic ndings and interpretations. Impacts, 2019, 21, 905 DOI: 10.1039/c9em90015b rsc.li/espi on 10 April 2019. Downloaded on 10/24/2024 11:52:23 A nsed under a Creative Commons Attribution 3.0 Unported John Pilewski,a Shikha Sharma,a Vikas Agrawal,a J. Alexandra Hakalab and Mengling Y. Stuckmanb Correction for ‘Eﬀect of maturity and mineralogy on ﬂuid-rock reactions in the Marcellus Shale’ by John Pilewski et al., Environ. Sci.: Processes Impacts, 2019, DOI: 10.1039/c8em00452h. In the original article, the %R0 values on the x-axis of Fig. 3C were incorrect. The Figure with the corrected values is shown below. The authors apologise for this error and state that this error does not aﬀect any of the scientic ndings and interpretations. Fig. 3 (C) Plot of the relationship between total target VOCs, total target organic acids, TOC, and %R0. The lines indicate the direction of the data trend and do not represent a calculated ﬁt of the data. Open Access Article. Published on 10 April 2019. D This article is licensed under a Creativ Fig. 3 (C) Plot of the relationship between total target VOCs, total target organic acids, TOC, and %R0. The lines indicate the direction of the data trend and do not represent a calculated ﬁt of the data. The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers. This journal is © The Royal Society of Chemistry 2019 Environ. Sci.: Processes Impacts, 2019, 21, 905 \| 905 Environ. Sci.: Processes Impacts, 2019, 21, 905 \| 905	39,170
https://openalex.org/W4297206378	OpenAlex	Open Science	CC-By	2,022	LEADERSHIP STRATEGIES ON PERFOMANCE IN COUNTY GOVERNMENT OF UASIN GISHU, KENYA	Jack Wainaina Kariuki	English	Spoken	11,920	19,772	ISSN 2350-1022 ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org Keywords: Working Environment, Organizational Performance. Keywords: Working Environment, Organizational Performance. LEADERSHIP STRATEGIES ON PERFOMANCE IN COUNTY GOVERNMENT OF UASIN GISHU, KENYA Jack Wainaina Kariuki1, Dr. Elizabeth Nambuswa Makokha1.2 , Dr. Duncan Nyaberi1.3 DOI: https://doi.org/10.5281/zenodo.7104938 DOI: https://doi.org/10.5281/zenodo.7104938 Published Date: 22-September-2022 Published Date: 22-September-2022 Published Date: 22-September-2022 Abstract: The purpose of this study was to investigate the effects of leadership strategies on performance of Uasin Gishu County. The study was guided by the following specific objectives; to evaluate the effect of work environment on performance in Uasin Gishu County. The study was guided by Transformational Leadership Theory. A descriptive survey research design was employed with a target population of 95 respondents consisting of the county executive committee members, county chief officers, county directors, chief principals, and principals. The data collection instrument was questionnaires both closed and open ended. Piloting was done to test the validity and reliability of the data collection instrument. Data collected using questionnaires was entered and analyzed using Statistical Package for Social Science (SPSS) software. In addition, inferential statistics that include Analysis of variance, Pearson correlation and multiple regression analysis was used establishing the nature and extent of relationship between variables. To ascertain the relationship between the independent variables and the dependent variable, a regression model was used. In conclusion basing on the findings, the study concluded that working environment has a significant effect on performance of Uasin Gishu county government. The study recommended that the management of the county government of Uasin Gishu should provide their employees a fair and conducive working condition to enable high performance. Due to an ever changing environment, the county government of Uasin Gishu should embrace regular trainings to enhance high and required competencies that facilitates good county government performance. The management of the county government of Uasin Gishu should establish a good relationship with their stakeholders through provision of good reception to enhance customer satisfaction. The study is useful to all the stakeholders in the County governments. Paper Publications Jack Wainaina Kariuki1, Dr. Elizabeth Nambuswa Makokha1.2 , Dr. Duncan Nyaberi1.3 1School of Business, Department of Business. Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000 - 00200, Nairobi Kenya 2College of Human Resource Development, Department of Entrepreneurship, procurement, leadership and management. .Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000 - 00200, Nairobi Kenya DOI: https://doi.org/10.5281/zenodo.7104938 1. INTRODUCTION The core purpose of leadership strategic theory and research is to understand how much influence top executives have over performance (Singh et al. 2016). Empirical and conceptual studies have shown that strategic leadership actions significantly influence performance (Quigley & Graffin, 2017; unfortunately, other studies conclude that their actions are impeded by situational constraints, inertia or random effects, such that they don’t have much leeway over performance (Fitza, 2017; Haveman, 1992). These divergent findings indicate either a lack of evidence in establishing a direct Page \| 37 Page \| 37 Paper Publications Paper Publications Leadership strategies are based on a thorough analysis of the current situation and an informed view of the future. The strategy then provides a series of recommendations to close the gap between the current situation and desired future. Once the leadership strategy is known, a leadership development strategy can be formulated to produce the desired future state, and implications for talent management processes can be identified. When the strategy is implemented, business results will provide feedback on how well the leadership strategy is working and help shape what new business strategies can be considered with the leadership talent that has been developed. In reality, the leadership strategy implications would be much more specific, reflecting the actual opportunities and issues surrounding the key drivers. The key drivers and their associated business strategies should have clear implications for what leaders must do well in order for the organization to succeed. Ultimately, leadership development activities should be designed to ensure that individual leaders and the collective leadership of the organization are prepared to implement the most important strategies related to the key drivers. The leadership strategies may also include specific target dates for each action, who is responsible, barriers to success, costs and implications for leadership development or talent management processes. Various scholars have argued that methodological and statistical limitations, unavailability of relevant control variables and contexts have systematically undermined the effect of strategic leadership on performance (Fitza, 2017; Hambrick & Quigley, 2014; Blettner et al., 2012). The world is increasingly getting complex, and so are local governments. With multiple dynamic and competing factors coming into play, there is need to put in place effective strategic management with good leadership plan to meet modern management challenges in counties .To compete successfully in this environment, counties continually need to improve their performance by reducing cost, innovating products and processes and improving quality, productivity and speed to market(Rumelt,2011). Strategic Competitive advantage is what keeps great organizations ahead of their competitors. Rothaermel (2012) pointed out that the organization which has a competitive advantage performs better financially than other competitors in the industry. Some companies may achieve it without a strategic plan but for most players, its vital to plan strategically (Rothmeyer, 2012) so are the counties since they’re like organizations. Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org association between strategic leadership and performance or of the many confounding variables that make it difficult to demonstrate a clear cause and effect (Knies et al., 2016). Hambrick & Quigley (2014) point out that scholars are yet to agree on the conceptualization and operationalization of contextual conditions strategic leaders face. A leadership strategy makes explicit how many leaders we need, of what kind, where, with what skills, and behaving in what fashion individually and collectively to achieve the total success we seek. A good leadership strategy takes all necessary factors into account. Simply having all of the leadership positions on the organization chart filled will not produce the leadership that is required to implement strategies, adapt to change, support innovation or other important organizational agendas. It is not just having the right number of bodies, it is what those bodies do and how they relate to one another that matters. association between strategic leadership and performance or of the many confounding variables that make it difficult to demonstrate a clear cause and effect (Knies et al., 2016). Hambrick & Quigley (2014) point out that scholars are yet to agree on the conceptualization and operationalization of contextual conditions strategic leaders face. A leadership strategy makes explicit how many leaders we need, of what kind, where, with what skills, and behaving in what fashion individually and collectively to achieve the total success we seek. A good leadership strategy takes all necessary factors into account. Simply having all of the leadership positions on the organization chart filled will not produce the leadership that is required to implement strategies, adapt to change, support innovation or other important organizational agendas. It is not just having the right number of bodies, it is what those bodies do and how they relate to one another that matters. Paper Publications Paper Publications Muasya (2017) the role of strategic leadership in effective strategy implementation a case study of UNICEF Somalia which illustrates very well good relationship between leadership strategies and proves that its very crucial in performance of an organization. Strategic Leadership is conceived as a process where one or more persons influence a group of persons to move in a certain direction. Messick & Krammer (2014) argued that the degree to which the individual exhibits leadership traits depends not only on his characteristics and personal abilities, but also on the characteristics of the situation and environment in which he finds himself. According to Pearce and Robinson (2014), strategic leadership is about coping with change; and more changes always demand more leadership. Hitt, Ireland and Hoskisson (2014) define strategic leadership as the leader’s ability to anticipate, envision, and maintain flexibility to empower others to create strategic change as necessary; it involves managing through others. Capon (2016) defines strategic leadership as the ability to influence a group towards the achievement of goals. He further states that good leadership has strategic vision and is persuasive at implementing strategy to achieve tangible results. Lynch (2015) is of the view that strategic leadership typically involves communicating with and listening to those inside the organization with the aim of spreading knowledge, creating and innovating new areas and solutions to problems. It is the process of allocating resources to support the chosen strategies. The African strategic leadership process includes the various management activities that are necessary to put strategy in motion, institutes strategic controls that monitor progress, and ultimately achieve organizational goals (Barnat, 2014). Thompson, Strickland and Gamble (2010) emphasizes that there can be no doubt that effective organizational leadership and the consistency of a strong organizational culture are two central ingredients in enabling successful execution of a firm’s strategies and objectives. Weak leadership can wreck the soundest strategy; forceful execution of even a poor plan can often bring victory. Strategy implementation may be said to consist of securing resources, organizing these resources and directing the use of these resources within and outside the organization (Mintzberg, 1994). Strategy implementation is only successful when it is backed by effective leadership. House & Mitchell (2014) mentions that effective leaders clearly specify the task, reduce road blocks to task achievement, and increase opportunities for task related satisfaction and improve performance. This clearly shows that strategic leadership is linked to organization performance. Paper Publications 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org spent mostly on resolving issues instead of analyzing the local and global environment. A strategic manager has a vision and a goal but that is hardly enough. Strategic global Leadership is creative, innovative and proactive (Sullivan, 2006). With the changing and highly competitive global business environment, it is paramount for the company to practice strategic leadership. Leaders anticipate troubles and opportunities. They are highly observant and aware of their surroundings and the changes therein. To a strategic leader flexibility means staying open to opportunities as they arise (Mazur, 2010). They observe the emerging trends and patterns and adjust to the global changing business climate. spent mostly on resolving issues instead of analyzing the local and global environment. A strategic manager has a vision and a goal but that is hardly enough. Strategic global Leadership is creative, innovative and proactive (Sullivan, 2006). With the changing and highly competitive global business environment, it is paramount for the company to practice strategic leadership. Leaders anticipate troubles and opportunities. They are highly observant and aware of their surroundings and the changes therein. To a strategic leader flexibility means staying open to opportunities as they arise (Mazur, 2010). They observe the emerging trends and patterns and adjust to the global changing business climate. Regionally strategic leadership is very important element in the running of an organization since its assist to meet the goals and objectives set by an organization hence helps in smooth running of the organization in general. Similarly, for an organization to survive in the market for a long time competitively it must embrace strategic leadership in totality. In Uganda, experts have historically established that good strategic leadership produces good management, performance, accountability of public money, public behavior and better outcomes of public services (Sullivan & Skelcher, 2017). Good strategic leaders in the county government should guide their county government in the proper direction. This means that their values, vision and purpose are for the good of the county government and its stakeholders. Ethical leaders epitomize the values, vision, and purpose of the organization and of the constituents, within an understanding of ethical ideals (Lewis, 2012). Paper Publications Paper Publications Strategic management activities transform the strategic plan into a system that provides strategic performance feedback to decision making and enables the plan to evolve and grow as requirements .Strategy Execution by competent leaders is basically synonymous with Strategy Management and amounts to the systematic implementation of a strategy (Hambrick et al, 2012). The strategic leader considers the benefits of achieving the strategy and finds the challenges worthwhile (Morebusiness. com Editors, 2007). There are four approaches that are used by strategic leaders (Whittington, 2002). The first approach is the classical theory where the objective of the leaders is profit making. Rational thinking is used to attain it. If a business is not profitable it is discarded. The second theory, evolutionary theory is different in that it does not focus on rationality alone. Globally the environment in which the business is operating in is a critical factor. A radical kind of thinking and flexibility is required in complex and dynamic environments. By firms operating strategically in the dynamic environment, they ensure continuity of the organization (Ansoff, 1985). Many scholars have argued classical approach is irrelevant since it does not put into consideration the current dynamic environments (Skaik, 2009). Strategic management ensures work is done. Managers focus on the daily directing and controlling activities. It is the authority and role given to them by the organization. The strategic managerial skills are a great asset though deficient in the current fast-paced changing environment globally (Coach4growth Editors, 2010). This is because strategic management is mostly reactive to situations. Non strategic manager globally do nothing to offset most of the future risk (CMOE Editors, 2010). They may even resist change instead of adapting to it and seeing the opportunities and competitive advantages therein. Strategic Managers end up doing a lot of fire-fighting and are not proactive. Energy is Page \| 38 Page \| 38 Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. Paper Publications Locally ,strategic leadership is more significant as in the case of Masungo, et al., (2015) who established the effect of strategic leadership on the performance of devolved government system in Kakamega County, Kenya whereby they noted that performance of devolved systems was dependent to the strategic leadership .Kitonga, et al.,(2016) who found that leadership strategies are very important in the overall success of the entire organization in general. Similarly Abba (2016) assessed the influence of strategic leadership in strategy implementation in commercial banks whereby he found out that for organizations to meet their long term strategic growth they must put up strategic leadership well in place always , Mutia (2015) did a study on strategic leadership and its influence on church growth in Kenya where he found out that organization should place an emphasis on training and development to upgrade the existing skill and knowledge base of the employees for effective performance Page \| 39 Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org In the year 2010, Kenya ushered in a highly ambitious form of government that sought to change the relationship between the government and citizens when the constitution was promulgated (COK 2010). The constitution saw shifting of government from centralized to decentralized, and from “top-down” to “bottom up”. Its emphasis is on strengthening public participation. It is a core element in Kenya’s strategy to accelerate growth and address long-standing inequalities in economic opportunities, investment, and service delivery in different parts of the country. Forty-seven county governments were created under Chapter eleven of Constitution of Kenya. The functions of county governments in Kenya came into existence in 2013 when the country elected its leaders under the new dispensation of devolution. The county governments are in charge of health care, pre-primary education and maintenance of local roads. They also, collect revenue in their areas of jurisdiction such as taxes on property and entertainment (Kimenyi, 2013). According to the constitution, only 15% of the government revenue is allocated to the counties whereas 85% remains in the central government, COK 2010. Mwangi (2013) states that each county gets its share (i.e. Paper Publications out of the 15%) based on an horizontal formula that was set by the Commission of Revenue Allocation (CRA) based on the weightings: population 45%, poverty index 20%, land area 8%, basic equal share 25%, fiscal responsibility 2%. Because each county is autonomous in determining on how to spend its financial resources in meeting its development agenda, there is need to assess their ability in managing resources for the purposes of effective service delivery to the people (GOK, 2013). The constitution also gives a provision for those counties that mobilize and manage their resources to be rewarded by receiving a higher share of the central government revenue. In Kenya, since independence the Government has been implementing local government reform programmes with the view of enhanced service delivery thus good performance but this effort have met several challenges. This is the baseline why this study seeks to investigate the effect of strategic leadership on the performance of county governments. The county government of Uasin Gishu is managed from the county head office at the former Municipal hall, its enjoys good old structures that have been in existence for a very long time and barely are new structures within the county head offices, which makes the county maintain its original heritage. The county government strategies are important such that they should be aligned with good competent leadership so as to achieve the county government objectives of service delivery to its locals according to local government devolution directions. The research done by Kipkorir (2013) on the factors influencing implementation of strategic plans in local authorities in Migori County suggested that organizational culture, structure, strategic leadership and financial resources affected the implementation of the strategic plan. The study conducted by Obiga (2014) focused on the challenges faced during the formulation and implementation of strategy within the County Government of Nairobi. Another study closely related to this was on the strategic planning and implementation practices in Kisii County conducted by Opano (2013) which suggested that poverty line and high mortality rate were some of the challenges faced by the County Government in implementing the strategic plans. Effective leadership Strategy execution is a process by which strategies are put into concrete action through improvement of programs, budgets, and actions by competent strategic managers. Paper Publications 2. EFFECT OF WORK ENVIRONMENT ON PERFORMANCE IN COUNTY GOVERNMENT OF UASIN GISHU, KENYA Workplace environment is the sum of the interrelationships that exists within the employees and the environment in which they work (Kohun, 2002). According to Heath (2006), this environment involves the physical location as well as the immediate surroundings, behavioral procedures, policies, rules, culture, resources, working relationships, work location, all of which influence the ways employees perform their work. The quality of the workplace environment impacts on employees‟ performance and subsequently influences the organization competiveness. An effective workplace environment management entails making work environment attractive, comfortable, satisfactory and motivating to employees so as to give employees a sense of pride and purpose in what they do (Humphries, 2005). Employees will and are always contented when they feel their immediate environment; both physical sensations and emotional states are in tandem with their obligations (Farh, 2012) and how well employees connect with their organization‟s immediate workplace environment, influences to a great extent their error rate levels, efficiency and innovativeness, collaboration with other employees, absenteeism and, ultimately their retention (Leblebici, 2012) The type of workplace environment in which employees operate determines whether or not such organizations‟ will prosper (Chandrasekhar, 2011). Physical workplace environment contextualizes the office layout and design while psychosocial factors include working condition, role congruity and social support from supervisors. Policies encompass employment conditions of employees derived from industrial instruments and agreements negotiated with employees and unions, along with our Working environment plays an important role towards the employees ‘performance. Working environment is argued to impact immensely on employees’ performance either towards negative or the positive outcomes (Chandrasekar 2011).In the world, there are international organizations who debate the rights of employee. Most people spend fifty percent of their lives within indoor environments, which greatly influence their mental status, actions, abilities and performance (Dorgan, 2014). Better outcomes and increased productivity is assumed to be the result of better workplace environment. Better physical environment of office will boosts the employees and ultimately improve their productivity. Various literature pertain to the study of multiple offices and office buildings indicated that the factors such as dissatisfaction, cluttered workplaces and the physical environment are playing a major role in the loss of employees’ productivity (Croome, 2017). The factors of work environment had changed due to the changes in several factors such as the social environment, information technology and the flexible ways of organizing work processes (Hasun & Makhbul, 2015). Paper Publications The process is often referred to as operational planning and usually comprise the allocation of day-to-day decisions in resource allocations Facts from the extant literature reveals that competitive advantages rooted in organizational internal competencies can be more safer in creating benefits for institutions, in contrast to its external opportunities (Arasa & K`obonyo, 2012). One of the organizational internal competitive superiorities is leadership style (Safarzadeh et al. 2015).Strategic leadership was hypothesized to have a positive impact on organizational innovativeness (Safarzadeh et al. 2015). Also, strategic leaders have been repetitively recognized for their decisive role in recognizing opportunities and taking positive decisions that will have impact on innovation process (Safarzadeh et al. 2015). More so, strategic leadership and organizational innovativeness are considered to be fundamentals for achieving and maintaining strategic competitiveness in the 21st century. For the successful implementation of strategies, the challenge of leadership is to be strong but not rude, to be good but not weak, to be humble but not shy, to be proud but not arrogant, to have humor but without foolishness. Strategy can also be explained as choosing one process or several processes to achieve the main long-term goals of the organization. It also includes courses of action and the allocation of resources needed to achieve the desired objectives. (Elkhdr, 2019). Strategy can be explained as choosing one process or several processes to achieve the main long-term goals of the organization. It also includes courses of action and the allocation of resources needed to achieve the desired objectives. (Elkhdr, 2019). Various scholars have argued that methodological and statistical limitations, unavailability of relevant control variables and contexts have systematically undermined the effect of strategic leadership on performance (Fitza, Page \| 40 Page \| 40 Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org 2017). Boal & Hooijberg (2001) observe that studies on strategic leadership are limited since many have used demographic variables as substitutes for moderating or mediating variables. Crossan et al., (2008) argue that many studies have excluded critical organizational and environmental variables that might moderate or mediate such a relationship. Paper Publications Additionally, empirical literature has solely examined the impact of strategic leadership at the micro levels without integrating both the micro and macro perspectives of leadership (Bornardi et al., 2018; Kim et al., 2014). The core purpose of strategic leadership theory and research is to understand how much influence top executives have over performance (Singh et al. 2016). As noted above the various researches done by scholars were done on already early established organizations which the researchers had previous knowledge and materials to refer to whereas my research is on county governments which are new institutions in Kenya with little or no previous history or materials in Kenya. Despite application of strategic leadership in county government there have been challenges of competency, qualifications, motivation, involvement, bureaucracies, relatable values, reception, clarity, service delivery, and efficiency in county government of Uasin Gishu. Therefore the study seeks to determine the effects of work environment on performance of county government in Uasin Gishu County, Kenya. Paper Publications 2. EFFECT OF WORK ENVIRONMENT ON PERFORMANCE IN COUNTY GOVERNMENT OF UASIN GISHU, KENYA When employees’ are physically and emotionally fit will have the desire to work and their performance outcomes shall be increased. In today’s competitive business environment, organizations can no longer afford to waste the potential of their workforce. There are key factors in the employee’s workplace environment that impact greatly on their level of motivation and performance. The workplace environment that is set in place impacts employee morale, productivity and engagement - both positively and negatively. It is not just coincidence that new programs addressing lifestyle changes, work/life balance, health and fitness - previously not considered key benefits - are now primary considerations of potential employees, and common practices among the most admired companies. In an effort to motivate workers, firms have Page \| 41 Page \| 41 Paper Publications Paper Publications A business organization must produce the right things and it must produce them using the fewest possible inputs if it is to have a strong organizational performance. Businesses typically try to perform well in a number of areas. First, they try to perform well financially. That is, they need to realize a good return on their investment. They need to add as much value as possible in their production process. Second, they try to perform well in terms of the market or non-financial performance. What this means is that they must appropriately manage the determinants that lead to excellent performance. The performance determinants comprise of continuous learning and growth, and efficient internal processes. Didier Noyé (2002) believes that the performance consists in achieving the goals that were given to you in convergence of enterprise orientations. In his opinion, performance is not a mere finding of an outcome, but rather it is the result of a comparison between the outcome and the objective. Unlike other authors, Didier Noyé considers that this concept is actually a comparison of the outcome and the objective. The author’s definition is far from clear, as both outcomes and objectives vary, most often, from one field of activity to another. Author Michel Lebas (1995) characterizes the performance as future-oriented, designed to reflect particularities of each organization / individual and is based on a causal model linking components and products. He defines a ʺsuccessfulʺ business as one that will achieve the goals set by the management coalition, not necessarily one that achieved them. Thus, performance is dependent as much of capability and future. Unlike other authors, Michel Lebas noted the difference between ʺa performanceʺ, ʺperformanceʺ and ʺbeing performantʺ. ʺA performanceʺ is subject generally to a measured result, higher than that provided for or arising from the previous results. ʺA performanceʺ thus indicates always a positive connotation. ʺPerformanceʺ can be both positive and negative and relates to past results. For Whooley (1996), performance is not an objective reality, waiting somewhere to be measured and assessed, but a socially constructed reality that exists in people’s minds, if it exists somewhere. According to the author, performance may include: components, products, consequences, impact and can also be linked to economy, efficiency, effectiveness, cost effectiveness or equity. Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org implemented a number of practices such as performance based pay, employment security agreements, practices to help balance work and family, as well as various forms of information sharing. In addition to motivation, workers need the skills and ability to do their job effectively. And for many firms, training the worker has become a necessary input into the production process. implemented a number of practices such as performance based pay, employment security agreements, practices to help balance work and family, as well as various forms of information sharing. In addition to motivation, workers need the skills and ability to do their job effectively. And for many firms, training the worker has become a necessary input into the production process. Employee performance is the combined result of effort, ability, and perception of tasks (Platt, 2010). Employees‟ performance is imperative for organizational outcomes and success. Many factors influence employee performance; and workplace environment factors stands out as the key determinants of performance. It is the key multi character factor intended to attain outcomes and has a major connection with planned objectives of the organization (Sabir et al. 2012). Favourable workplace environment guarantees the wellbeing of employees as well as enables them to exert themselves to their roles with all energy that may translate to higher performance (Taiwo, 2010). To survive and grow, institutions have to embrace strategies of keeping the quality of the workplace high as a competitive strategy. Since employees are the key resource and greatest expense of any institution, the long-term benefits of a properly designed and user-friendly workplace environment should be factored into any initial cost considerations (Smith, 2011) Thus work environment is key in performance of an organization. The concept of organizational performance is connected to the ideas of effectiveness and efficiency (Kaplan & Norton, 2016). Paper Publications Paper Publications Both Lebas (1995) and Whooley (1996) consider performance as subjective and interpretative, not least, being related to the cost lines, which emphasizes the ambiguous nature of the concept. Neely (2002) believes that performance should consider quantifying the efficiency and effectiveness of actions. This quantification can be expressed both qualitatively and quantitatively. According to the definition of Neely and other authors, performance is closely related to efficiency and effectiveness. Kane (1996) argues that the performance is ʺsomething that a person leaves behind and which exists outside the said purposeʺ. According to Kane, performance is defined at the level of each individual within the organization or at organization level. It is perceived as an understanding of the achieved results. The author emphasizes the particular nature of the definition and the impossibility of outlining a general definition. Therefore, we can speak of an accuracy of the definition at particular level and an ambiguity of it at general level. Page \| 42 Paper Publications Page \| 42 Page \| 42 Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org Traditionally most of the managerial performance measures have been based on financial measures of performance (Mishra and Mohanty, 2014). Also, it is mentioned that these measures include return on assets (ROA), return on equity, and return on sales (Mishra and Mohanty, 2014). In order to define the firm’s performance it is important to mention that “financial indicators reflect the fulfillment of a multinational enterprises’ (MNE’s) economic goals in financial terms (Richter et al., 2017). Also, “market-based financial indicators take the investor’s view (e.g. cumulative abnormal returns, market-to-book ratio, excess value) and concentrate on or involve risk considerations (Richter et al., 2017). According to Richter et al. (2017) all these indicators form “the narrowest conception of business performance and the border conceptualization of performance is operational performance”. In this context it is necessary to mention the following aspects: -“operational indicators focus on those key determinants of success that might lead to financial performance (Richter et al., 2017); -effectiveness measures include reputation, survival, achievement of goals, performance in relation to competitors, etc. 3. METHOD A descriptive survey research design was employed with a target population of 95 respondents consisting of the county executive committee members, county chief officers, county directors, chief principals, and principals. The data collection instrument was questionnaires both closed and open ended. Piloting was done to test the validity and reliability of the research instrument. Piloting was done to test the validity and reliability of the data collection instrument. Data collected using questionnaires was entered and analyzed using Statistical Package for Social Science (SPSS) software. In addition, inferential statistics that include Analysis of variance, Pearson correlation and multiple regression analysis was used establishing the nature and extent of relationship between variables. To ascertain the relationship between the independent variables and the dependent variable, a regression model was used. Paper Publications (Richter et al., 2017 ); -operational effectiveness, organizational effectiveness and financial performance (indicators) are interrelated” (Richter et al., 2017); -organizational effectiveness performance is supposed to be reflected in better financial performance in the medium or long term, because performance improvement is at the heart of strategic management and organization theory” (Venkatraman and Ramanujam 1986; Ginsberg and Venkatraman, 1985; in Richter et al., 2017); -financial performance influences organizations and operational and organizational performance (Richter et al., 2017). Finally they strategic leader who are vital managers must strive to achieve organizational effectiveness by, quality service delivery, customer satisfaction and creation of a conducive environment for service delivery. In addition, they need to perform well in terms of creating customer royalty and building trust. According to (Lehner, 2014) Non-financial performance is by assessing the activities that an organization sees as important to the achievement of its strategic objectives. Typical non financial measures include; measures that relate to customer relationships, employees, operations, quality, cycle-time, and the organization’s supply chain or its pipeline. Paper Publications 4. DISCUSSION Workplace environment is the sum of the interrelationships that exists within the employees and the environment in which they work (Kohun, 2002). According to Heath (2006), this environment involves the physical location as well as the immediate surroundings, behavioral procedures, policies, rules, culture, resources, working relationships, work location, all of which influence the ways employees perform their work. Employees will and are always contented when they feel their immediate environment; both physical sensations and emotional states are in tandem with their obligations (Farh, 2012) and how well employees connect with their organization’s immediate workplace environment, influences to a great extent their error rate levels, efficiency and innovativeness, collaboration with other employees, absenteeism and, ultimately their retention (Leblebici, 2012) The type of workplace environment in which employees operate determines whether or not such organizations‟ will prosper (Chandrasekhar, 2011). The study sought to determine the effects of work environment on performance of county government in Uasin Gishu County, Kenya. The findings are presented in a five point Likerts scale where SA=strongly agree, A=agree, N=neutral, D=disagree, SD=strongly disagree and T=total. Results from table 1 below on whether having good working practices facilitates the performance of county government revealed that 43.0 percent of the respondents strongly agreed, 35.0 percent of them agreed, 1.0 percent of the respondents were neutral, 16.0 percent disagreed while 4.0 percent of them strongly disagreed. These findings implied that having good working practices facilitates the performance of county government. Page \| 43 Page \| 43 Paper Publications ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org Following closely on the issue of attitude was the question of whether relatable values in a working environment facilitates a great performance of county government. The distribution of the responses indicated that 33.0 percent strongly agreed to the statement, 31.0 percent of them agreed, 2.0 percent of them were neutral, 18.0 percent of them disagreed while 16.0 percent of them strongly disagreed to the statement. These findings implied that relatable values in a working environment facilitates a great performance of county government. The respondents were also asked whether a supportive atmosphere as an effective working environment facilitates County government performance. 4. DISCUSSION The distribution of the responses indicated that 35.0 percent strongly agreed to the statement, 32.0 percent of them agreed, none of them were neutral, 18.0 percent of them disagreed while 16.0 percent of them strongly disagreed to the statement. These findings implied that a supportive atmosphere as an effective working environment facilitates County government performance. The respondents were further asked whether a great working environment is contributed much more by the style of leadership in county government. The distribution of the responses indicated that 37.0 percent strongly agreed to the statement, 28.0 percent of them agreed, 5.0 percent of them were neutral while 20.0 percent and 10.0 percent of them disagreed strongly and disagreed to the statement respectively. These findings implied that a great working environment is contributed much more by the style of leadership in county government. : Effect of work environment on performance of county government in Uasin Gishu County, Kenya Table 1: Effect of work environment on performance of county government in Uasin Gishu County, Kenya Statements SA A N D SD Having good working practices facilitates the performance of county government % 43.0 35.0 1.0 16.0 4.0 Relatable values in a working environment facilitates a great performance of county government % 33.0 31.0 2.0 18.0 16.0 A supportive atmosphere as an effective working environment facilitates County government performance % 35.0 32.0 0.0 18.0 16.0 A great working environment is contributed much more by the style of leadership in county government % 37.0 28.0 5.0 20.0 10.0 4.1.2 Multiple Linear Regression Multiple linear regressions were computed at 95 percent confidence interval (0.05 margin error) to show the multiple linear relationship between the independent and dependent variables of the study. International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org 4.1.2.3 Regression Coefficients From the Coefficients table (Table 4.5) the regression model can be derived as follows: 4.1.2.1 Coefficient of Determination (R2) Table 3 shows that the coefficient of correlation (R) is positive 0.476. This means that there is a positive correlation between the effects of leadership strategies on performance of Uasin Gishu county government, Kenya. The coefficient of determination (R Square) indicates that 23.6% of performance of Uasin Gishu county government, Kenya is influenced by leadership strategies. The adjusted R2 however, indicates that 19.5% of leadership strategies is influenced by the performance of Uasin Gishu county government, Kenya leaving 80.5% to be influenced by other factors that were not captured in this study. Table 3: Model Summary Model R R Square Adjusted R Square Std. Error of the Estimate 1 .476a .236 .195 .90429350 a. Predictors: (Constant), Working Environment 4.1.2.2 Analysis of Variance Table 4 shows the Analysis of Variance (ANOVA). The p-value is 0.000 which is < 0.05 indicates that the model is statistically significant in predicting leadership strategies affect performance of Uasin Gishu county government. The results also indicate that the independent variables are predictors of the dependent variable. The ANOVA results indicate that the independent variables significantly (F=7.841, P=0.000) explain the variance of performance of Uasin Gishu county government. Table 4 ANOVAa Model Sum of Squares df Mean Square F Sig. 1 Regression 246.214 4 4.712 7.841 .000b Residual 77.050 80 .832 Total 313.264 85 A. Dependent Variable: Performance of Uasin Gishu County Government B. Predictors: (Constant), Working Environment 4 1 2 3 R i C ffi i t Table 4 ANOVAa 4.1.1 Pearson Correlation The study sought to establish the strength of the relationship between independent and dependent variables of the study. Pearson correlation coefficient was computed at 95 percent confidence interval (error margin of 0.05). Table 2 illustrates the findings of the study. Table 2: Correlation Matrix Performance of Uasin Gishu County Government Working environment Pearson Correlation .899** Sig. (2-tailed) .000 N 85 Results as shown on Table 2 above, the p-value for working environment was found to be 0.000 which is less than the significant level of 0.05, (p<0.05). The result indicated that Pearson Correlation coefficient (r-value) of 0.899, which represented a strong, positive relationship between working environment on performance of Uasin Gishu county government. Table 2: Correlation Matrix Results as shown on Table 2 above, the p-value for working environment was found to be 0.000 which is less than the significant level of 0.05, (p<0.05). The result indicated that Pearson Correlation coefficient (r-value) of 0.899, which represented a strong, positive relationship between working environment on performance of Uasin Gishu county government. Results as shown on Table 2 above, the p-value for working environment was found to be 0.000 which is less than the significant level of 0.05, (p<0.05). The result indicated that Pearson Correlation coefficient (r-value) of 0.899, which represented a strong, positive relationship between working environment on performance of Uasin Gishu county government. Results as shown on Table 2 above, the p-value for working environment was found to be 0.000 which is less than the significant level of 0.05, (p<0.05). The result indicated that Pearson Correlation coefficient (r-value) of 0.899, which represented a strong, positive relationship between working environment on performance of Uasin Gishu county government. Page \| 44 Paper Publications Page \| 44 ISSN 2350-1022 4.1.3 Hypothesis Testing Ho1: Working environment does not have a significant effect on performance of Uasin Gishu county government. From Table 5 above, working environment (β = .598) was found to be positively related performance of Uasin Gishu county government. From t-test analysis, the t -value was found to be 3.571 and the ρ -value 0.000. Statistically, this null hypothesis was rejected because ρ<0.05. Thus, the study accepted the alternative hypothesis and it concluded that working environment affect performance of Uasin Gishu county government. 5. CONCLUSION AND RECOMMENDATION In conclusion basing on the findings, the study concluded that working environment has a significant effect on performance of Uasin Gishu county government. The study recommended that the management of the county government of Uasin Gishu should provide their employees a fair and conducive working condition to enable high performance. Due to an ever changing environment, the county government of Uasin Gishu should embrace regular trainings to enhance high and required competencies that facilitates good county government performance. The management of the county government of Uasin Gishu should establish a good relationship with their stakeholders through provision of good reception to enhance customer satisfaction. International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org = 0.079+ 0.598X4 The results in table 5 indicate that all the independent variables have a significant positive effect on performance of Uasin Gishu county government. The working environment with a coefficient of 0.598 (p-value = 0.000) influences performance of Uasin Gishu county government. According to this model when all the independent variable values are zero; performance of Uasin Gishu county government of will have a score of 0.079. Table 5: Coefficientsa Model Unstandardized Coefficients Standardized Coefficients t Sig. B Std. Error Beta 1 (Constant) .079 .091 .019 .000 Working environment .598 .105 .393 3.571 .000 A. Dependent Variable: Performance of Uasin Gishu County Government Page \| 45 Paper Publications Table 5: Coefficientsa Model Unstandardized Coefficients Standardized Coefficients t Sig. B Std. Error Beta 1 (Constant) .079 .091 .019 .000 Working environment .598 .105 .393 3.571 .000 A. Dependent Variable: Performance of Uasin Gishu County Government Table 5: Coefficientsa A. Dependent Variable: Performance of Uasin Gishu County Government ISSN 2350-1022 International Journal of Recent Research in Mathematics Computer Science and Information Technology Vol. 9, Issue 1, pp: (37-53), Month: April 2022 – September 2022, Available at: www.paperpublications.org REFERENCES [1] Adoli, H. L., & Kilika, J. M. (2020). Conceptualizing the role of leadership strategy in the context of strategic management process: A review of literature. Journal of Economics and Business, 3(4). [2] Al Shobaki, M. 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https://openalex.org/W3112128482_1	Spanish-Science-Pile	Open Science	Various open science	2,021	Impacto del uso del fideicomiso como alternativa de financiamiento en la rentabilidad de las principales empresas agroexportadoras de Lima por el año 2018	None	Spanish	Spoken	7,840	14,147	Impacto del uso del fideicomiso como alternativa de financiamiento en la rentabilidad de las principales empresas agroexportadoras de Lima por el año 2018 Item Type info:eu-repo/semantics/bachelorThesis Authors Gomez Perez, Michelle Valerie Romina; Orrego Ruiz, José Manuel Publisher Universidad Peruana de Ciencias Aplicadas (UPC) Rights info:eu-repo/semantics/openAccess Download date 28/07/2024 20:30:42 Item License http://creativecommons.org/licenses/by-nc-sa/4.0/ Link to Item http://hdl.handle.net/10757/652542 UNIVERSIDAD PERUANA DE CIENCIAS APLICADAS FACULTAD DE NEGOCIOS PROGRAMA ACADÉMICO DE CONTABILIDAD Y ADMINISTRACIÓN IMPACTO DEL USO DEL FIDEICOMISO COMO ALTERNATIVA DE FINANCIAMIENTO EN LA RENTABILIDAD DE LAS PRINCIPALES EMPRESAS AGROEXPORTADORAS DE LIMA POR EL AÑO 2018 TESIS Para optar el título profesional de Licenciado en Contabilidad AUTOR(ES) Gomez Perez, Michelle Valerie Romina (0000-0001-6764-0529) Orrego Ruiz, José Manuel (0000-0002-1122-390X) ASESOR Herrera Guillén, David Alberto (0000-0002-5503-6348) Lima, 08 de febrero de 2020 I AGRADECIMIENTOS El presente trabajo ha sido elaborado con esfuerzo por nuestra parte durante todo el largo proyecto de esta investigación. Agradecer a nuestras familias quienes fueron un gran pilar para la construcción de nuestra etapa profesional y el gran apoyo que nos brindaron. De igual manera, a nuestro estimado asesor quien nos guio durante todo este tiempo y nos permitió concluir satisfactoriamente con esta investigación académica. II RESUMEN La realización de la presente investigación tiene como principal finalidad definir y evaluar el impacto del uso del fideicomiso como herramienta financiera en las principales empresas del sector agroexportador durante el periodo del 2018. La investigación se basa en la búsqueda de una alternativa de financiamiento para las agroexportadoras. El financiamiento es un factor relevante para la expansión e inversión constante de cualquier tipo de negocio; sin embargo, con el fin de reducir el riesgo de cobranzas, las entidades financieras deciden realizar evaluaciones a sus solicitantes de los servicios de subvención de acuerdo con el monto pactado. La herramienta financiera más usual en Perú es el préstamo bancario tradicional y las tasas de interés son impuestas por los bancos después de evaluar al sector. En el caso de las agroexportadoras, considerando su producción estacional y la amplia cantidad de factores de riesgo inherentes al rubro, los bancos cobran una tasa más cara. El fideicomiso es una herramienta financiera muy poco conocida en el mercado; sin embargo, presenta tendencias de crecimiento de demanda en los últimos años. Consiste en ofrecer el mismo capital a una menor tasa con otras condiciones de cobranzas y una evaluación más específica para la entidad solicitante. En el siguiente trabajo realizaremos dicha evaluación y comparación entre el tipo de financiamiento más usual en Perú y el fideicomiso con el fin de demostrar la efectividad de este y la posibilidad de considerarlo como una alternativa financiera rentable. Como resultado de la investigación se validaron las hipótesis satisfactoriamente y se obtuvieron una conclusión acertada. Palabras clave: Fideicomiso, sector agroexportador, financiamiento, tasa de interés, fiduciario, bien fideicometido, riesgo inherente. NIIF 9. III ABSTRACT The present investigation has as its main purpose to define and evaluate the impact of the use of the escrow as a financial tool in the main companies of the agro-export sector during the period of 2018. The investigation is based on the search for a financing alternative for the agroexport companies, due to their production volatility, there are higher interest rates impounded by the financial entities. Financing is a relevant factor for the constant expansion and inversion of any type of business, however, in order to reduce the risk of collection, the financial entities decide to carry out evaluations on their applicants for the subvention services according to the amount agreed. The most common financial tool in Peru is the traditional banking service and the restaurants of interest are being impeached by banks after evaluating the sector. In the case of agro-exports, considering their seasonal production and the wide range of risk factors inherent to the red, the banks charge a more expensive price. The escrow is a financial tool very well known in the market, however, it presents trends of demand growth in the last years. It consists in offering the capitalism to a smaller restaurant with other conditions of payment and a more specific evaluation for the requesting entity. In the next work we will carry out a evaluation and comparison between the most usual type of financing in Peru and the escrow with the purpose of demonstrating the effectiveness of this and the possibility of considering it as a profitable financial alternative. As a result of the investigation, the hypotheses were satisfactorily validated, and a correct conclusion was reached. Keywords: Trust, agro-export sector, financing, interest rate, fiduciary, escrow-asset, inherent risk. IFRS IV INDICE INTRODUCCIÓN ................................................................................................................. 6 Capítulo I. Marco Teórico..................................................................................................... 7 1.1 Antecedentes: ......................................................................................................... 7 1.1.1 Incremento en la actividad agroexportadora en el 2018: ................................... 7 1.1.2 Riesgo del sector Agrícola: ................................................................................ 9 1.1.3 Uso Fideicomiso en la Gestión Financiera ...................................................... 10 1.2 El Fideicomiso ..................................................................................................... 12 1.2.1 Características del Fideicomiso ....................................................................... 14 1.2.2 Partes del Fideicomiso ..................................................................................... 17 1.2.3 Tipos de Fideicomiso ....................................................................................... 19 1.2.4 Contexto del Fideicomiso en el Perú ............................................................... 28 1.3 Sector Agrícola y Agroexportador ....................................................................... 30 1.3.1 Definición y Características del sector ............................................................ 30 1.3.2 Principales Riesgos del Sector Agrícola: ......................................................... 36 1.3.3 Amenazas del Sector Agrícola ......................................................................... 39 1.3.4 Agentes reguladores en el Sector Agrícola: ..................................................... 40 1.4 Evaluación de Financiamiento ............................................................................. 41 1.4.1 Tipos de Financiamiento .................................................................................. 43 1.4.2 Instrumentos de Financiamiento ...................................................................... 46 1.4.3 El financiamiento en el Sector Agroexportador............................................... 47 1.4.4 Indicadores de Rentabilidad ............................................................................. 48 1.5 Tratamiento Contable del Fideicomiso ................................................................ 51 1.5.1 Transferencia de los activos en fideicomiso .................................................... 52 1.5.2 Inversiones y gastos efectuados por el Fideicomiso ........................................ 53 1.5.3 Ingresos periódicos generados por el fideicomiso ........................................... 53 1.5.4 NIIF 9 – Instrumentos Financieros .................................................................. 54 Capítulo II. Plan de Investigación ........................................................................................ 57 2.1. El Problema.......................................................................................................... 57 V 2.1.1. Problema General............................................................................................. 57 2.1.2. Problema Especifico ........................................................................................ 58 2.2. Hipótesis .............................................................................................................. 60 2.2.1. Hipótesis General ............................................................................................. 60 2.2.2. Hipótesis Específica ......................................................................................... 60 2.3. Objetivos .............................................................................................................. 61 2.3.1. Objetivo General .............................................................................................. 61 2.3.2. Objetivo Específico .......................................................................................... 61 Capítulo III. Metodología de trabajo ................................................................................... 63 3.1 Tipo de Investigación ........................................................................................... 63 3.2. Diseño de la Investigación ................................................................................... 65 3.3 Investigación Cualitativa ..................................................................................... 66 3.3.1. Herramienta de recopilación de información ................................................... 66 3.3.2. Tamaño de la muestra cualitativa .................................................................... 68 3.4. Investigación Cuantitativa ................................................................................... 70 3.4.1. Instrumento de recolección de datos cuantitativos .......................................... 71 3.4.2. Determinación de la Población y Muestra ....................................................... 74 3.4.3. Proceso de recaudación de información cuantitativa ....................................... 78 Capítulo IV. Desarrollo de la Investigación ........................................................................ 80 4.1. Aplicación de instrumentos.................................................................................. 80 4.1.1. Evaluación de Marco Teórico .......................................................................... 80 4.1.2. Aplicación de Instrumentos: Entrevista a Profundidad ................................... 83 4.1.3. Aplicación de instrumentos: Encuestas ........................................................... 99 4.2 Caso Práctico ........................................................................................................... 114 4.2.1 Introducción: .................................................................................................. 114 4.2.2 Objetivo: ........................................................................................................ 114 4.2.3 Desarrollo:...................................................................................................... 114 Capítulo V. Análisis de Resultados ................................................................................... 125 VI 5.1. Aplicación de resultados .................................................................................... 125 5.1.1. Resultados del estudio Cualitativo ................................................................. 125 5.1.2. Resultados del estudio Cuantitativo ............................................................... 136 5.1.3. Análisis del caso práctico............................................................................... 143 Capítulo VI. Conclusiones y Recomendaciones ................................................................ 146 6.1. Conclusiones ...................................................................................................... 146 6.2 Recomendaciones .............................................................................................. 150 BIBLIOGRAFÍA ............................................................................................................... 155 ANEXOS ........................................................................................................................... 160 II. Diseño de Encuesta ............................................................................................ 161 III. Relación de Empresas Agroexportadoras 2018 ................................................. 163 IV. Cronograma de Actividades:.............................................................................. 169 V. Matriz de Consistencia:...................................................................................... 170 VII INDICE DE FIGURAS Figura 1. Ranking de producto agrícolas peruanos exportados ...................................................... 8 Figura 2. Estructura fideicomiso de activos .................................................................................. 20 Figura 3. Estructura fideicomiso de flujos .................................................................................... 20 Figura 4. Estructura de fideicomiso mixto.................................................................................... 21 Figura 5. Estructura de fideicomiso de Escrow – Condicionado .................................................. 22 Figura 6. Estructura del fideicomiso de puente inmobiliario........................................................ 23 Figura 7. Estructura del fideicomiso de proyectos inmobiliarios ................................................. 24 Figura 8. Estructura del fideicomiso control de flujos - Carta Fianza .......................................... 25 Figura 9. Estructura del fideicomiso de acuerdo de accionistas ................................................... 25 Figura 10. Estructura de fideicomiso de patrimonio familiar ....................................................... 26 Figura 11. Estructura de fideicomiso de gestión de deuda ........................................................... 27 Figura 12. Participación del Mercado Fiduciario 2018 ................................................................ 29 Figura 13. Cadena productiva del Sector Agrario - Exportación.................................................. 32 Figura 14. Cadena productiva del Agronegocio ........................................................................... 33 Figura 16. Exportaciones Agropecuarios y sus proyecciones al 2021.......................................... 34 Figura 17. Informe del producto bruto interno 2018 .................................................................... 35 Figura 18. Informe del PBI 2018 según actividad económica ...................................................... 35 Figura 19. Estimación de ocurrencia del Fenómeno del Niño-Niña ............................................. 36 Figura 20. Precio Internacional de Alimentos Básicos ................................................................. 37 Figura 21. Boletines Técnicos de plagas y enfermedades en las cosechas ................................... 37 Figura 22. Creación del banco de segundo piso - MiAgro ........................................................... 38 Figura 23. Potencial Fuente de crecimiento en el sector Agrario ................................................. 38 Figura 24. Vinculación del crédito y PBI en el sector Agropecuario. .......................................... 42 Figura 25. Esquema de la Metodología de investigación académica ........................................... 63 Figura 26. Formula de selección de muestra. ............................................................................... 77 Figura 27. Respuestas de la pregunta nro. 1 - Elaboración propia. ............................................ 100 Figura 28.Respuestas de la pregunta nro. 2 - Elaboración propia. ............................................. 101 Figura 29. Respuestas de la pregunta nro. 3 - Elaboración propia. ............................................ 101 Figura 30. Respuestas de la pregunta nro. 4 - Elaboración propia. ............................................ 102 2 Figura 31. Respuestas de la pregunta nro. 5 - Elaboración propia. ............................................ 103 Figura 32. Respuestas de la pregunta nro. 6 - Elaboración propia. ............................................ 103 Figura 33. Respuestas de la pregunta nro. 7 - Elaboración propia. ............................................ 104 Figura 34. Respuestas de la pregunta nro. 8 - Elaboración propia. ............................................ 105 Figura 35. Respuestas de la pregunta nro. 9 - Elaboración propia. ............................................ 105 Figura 36. Respuestas de la pregunta nro. 10 - Elaboración propia. .......................................... 106 Figura 37. Respuestas de la pregunta nro. 11 - Elaboración propia. .......................................... 107 Figura 38. Respuestas de la pregunta nro. 12 - Elaboración propia. .......................................... 107 Figura 39. Respuestas de la pregunta nro. 13 - Elaboración propia. .......................................... 108 Figura 40. Respuestas de la pregunta nro. 14 - Elaboración propia. .......................................... 109 Figura 41. Respuestas de la pregunta nro. 15 - Elaboración propia. .......................................... 109 Figura 42. Respuestas de la pregunta nro. 16 - Elaboración propia. .......................................... 110 Figura 43. Respuestas de la pregunta nro. 17 - Elaboración propia. .......................................... 111 Figura 44. Respuestas de la pregunta nro. 18 - Elaboración propia. .......................................... 111 Figura 45. Respuestas de la pregunta nro. 19 - Elaboración propia. .......................................... 112 Figura 46. Respuestas de la pregunta nro. 20 - Elaboración propia. .......................................... 113 3 INDICE DE TABLAS Tabla 1. Ranking de las 10 primeras empresas Agroexportadoras 2018. ..................................... 33 Tabla 2. Preguntas del objetivo general seleccionadas para entrevista. ....................................... 67 Tabla 3. Preguntas del objetivo específico 1 seleccionadas para la entrevista. ............................ 67 Tabla 4.Preguntas del objetivo especifico 2 seleccionadas para la entrevista. ............................. 68 Tabla 5. Sumilla del especialista sobre fideicomisos.................................................................... 69 Tabla 6. Sumilla del especialista sobre Agroexportadoras ........................................................... 69 Tabla 7. Sumillas del especialista sobre Banca financiera ........................................................... 70 Tabla 8. Preguntas del objetivo general seleccionadas para la encuesta. ..................................... 71 Tabla 9. Preguntas seleccionadas para la encuesta según objetivos y sus dimensiones. .............. 72 Tabla 10. Relación de principales empresas agroexportadoras según valor FOB US$. ............... 75 Tabla 11. Empresas agroexportadoras con oficinas en Lima por el año 2018. ............................ 75 Tabla 12. Tabla de variables de la formula de muestra. ............................................................... 77 Tabla 13. Empresas seleccionadas como muestra de investigación. ............................................ 78 Tabla 14. Escala de Likert utilizada para la encuesta. .................................................................. 79 Tabla 15. Extracto Teórico por Hipótesis General – Elaboración propia. .................................... 81 Tabla 16. Extracto Teórico por Hipótesis Especifica 1 – Elaboración propia. ............................. 82 Tabla 17. Extracto Teórico por Hipótesis Especifico 2 – Elaboración propia.............................. 83 Tabla 18. Respuestas de la encuesta realizada a empresas agroexportadoras. ............................. 99 Tabla 19. Números de proyectos evaluados - Caso Práctico ...................................................... 114 Tabla 20. Datos del caso practico Proyecto. 2 - Opción #1 [OG] .............................................. 115 Tabla 21. Flujo de caja del fideicomiso - Proyecto. 2 [OE2] ..................................................... 115 Tabla 22. Calculo de la TEA....................................................................................................... 116 Tabla 23. Datos del caso práctico Proyecto. 2 - Opción #2 [OG] .............................................. 121 Tabla 24. Flujo de caja del préstamo bancario - Proyecto. 2 ...................................................... 121 Tabla 25. Comparación de Tasas Fideicomiso Vs Préstamo Bancario ...................................... 123 Tabla 26. Comparación de tasas Fideicomiso vs Prestamo bancario ........................................ 124 Tabla 27. Resumen de respuestas de expertos - Objetivo General ............................................. 134 Tabla 28. Resumen de respuestas de expertos - Objetivo especifico 1 ...................................... 135 Tabla 29. Resumen de respuestas de expertos - Objetivo especifico 2 ...................................... 136 4 Tabla 30. Resumen de procesamiento de casos .......................................................................... 137 Tabla 31. Estadística de Fiabilidad sobre el instrumente – Alfa de Cron Bach.......................... 137 Tabla 32. Tabla cruzada Uso del Fid vs Financiamiento Emp. Agroexportadoras .................... 138 Tabla 33. Prueba de Chi-cuadrado Uso del fid y Rentabilidad emp. Agroexportadoras. ........... 139 Tabla 34. Tabla cruzada D1: Tipo de Fideicomiso (Activos)Y: Rentabilidad Emp Agroexp .. 140 Tabla 35. Prueba de Chi-cuadrado Tipo de FideicomisoRentabilidad Emp Agroexp. ............. 140 Tabla 36. Tabla cruzada D2: Normativa ContableY: Rentabilidad Emp Agroexp .................. 141 Tabla 37. Prueba de Chi-cuadrado Normativa ContableRentabilidad Emp. Agroexp. ............ 142 Tabla 38. Comparativo de tasas por opciones evaluadas en el caso practico – Proyecto. 1. ...... 143 Tabla 39. Comparativo de tasas por opciones evaluadas en el caso practico – Proyecto. 2. ...... 143 5 INTRODUCCIÓN Toda organización busca su rentabilidad y a largo plazo la expansión y desarrollo de proyectos. Es entonces cuando los directivos evalúan distintas formas de obtener el capital requerido para tal inversión: ya sea por un ahorro interno de la empresa o un financiamiento externo que es lo más usual. Se deben considerar distintos puntos de vista, ventajas y desventajas de cada herramienta financiera para el propósito de la organización, puntos tales como los presupuestos para reconocer montos solicitados de inversión y los gastos financieros por intereses, así como la periodicidad de los pagos para realizar el contraste en los futuros beneficios económicos. Cada sector tiende a buscar su crecimiento, para tales motivos las entidades buscan fuentes de financiamiento en sus inversiones, sin embargo, uno de los medidores sobre la línea crediticia para la solicitud de los prestamos bancarios es el riesgo inherente a los rubros. El 29 de agosto del 2019, el Diario Gestión publicó una noticia en referencia a la nueva Reestructuración de Fondo Agro Perú para el 2020 por parte del Poder Ejecutivo, proyecto de ley donde se asignaron S/. 440 millones para el desarrollo de las tierras agrícolas en Perú siendo el Agro Banco el encargado de su administración desde el 2009. Con esta noticia se hace más atractivo y relevante un sector fundamental en la economía peruana, la agricultura. Por ende, habrá mayores inversiones en este giro de negocio y las organizaciones agroexportadoras necesitaran capital para poder desarrollar proyectos de expansión. (Redacción Gestión, 2019) Comúnmente las organizaciones optan por un financiamiento tradicional, ya sea por un préstamo bancario, hipotecas o acciones debido a estrategias de mercado adoptada por las instituciones financieras. En el presente trabajo presentaremos al fideicomiso, una opción de herramienta de financiamiento que evaluaremos frente a otras alternativas y concluiremos cual es la más optima para el sector agroexportador. Desarrollaremos el impacto que tiene el fideicomiso en las empresas agroexportadoras de Lima-Perú en el año 2018 y nos explayaremos en compararlo con las anteriores opciones financieras a través de indicadores y ratios y el tratamiento contable que se aplica para este proceso. 6 Capítulo I. Marco Teórico En el presente Capítulo definiremos los principales conceptos que abarca nuestro tema de investigación y sus derivaciones que se encuentren enlazados necesariamente con nuestra disyuntiva. A continuación, se presentará los antecedentes encontrados sobre nuestra problemática en investigación. 1.1 Antecedentes Como investigaciones previas sobre nuestro tema hemos podido encontrar diversas fuentes que nos permitieron llegar al planteamiento de nuestra problemática y el contexto en que se encuentra. Cabe mencionar, que no se hallaron investigaciones donde se formulen nuestras variables de manera conjunta, lo cual resalta la justificación y relevancia de nuestra presente investigación. Por esta razón, con respecto a nuestra problemática seleccionamos tres aspectos sobre Investigaciones del Incremento en la actividad agroexportadora en el 2018, Investigaciones sobre el riesgo del sector Agrícola e Investigaciones del Fidecomiso como Garantía. 1.1.1 Incremento en la actividad agroexportadora en el 2018 En el Perú en los últimos 5 años el crecimiento económico se ha venido paralizando por distintos factores, ello no ha impedido el crecimiento del país, pero si al ritmo del crecimiento porcentual. Es necesario mencionar, que según Informe Anual 2018 del Desenvolvimiento del comercio exterior Agroexportador realizado por Servicios al Exportador (SIICEX) la agroexportación en el país se encuentra en su potencial crecimiento debido a que actualmente el mundo demanda alimentos con nuevas presentaciones organizas y exóticas. (SIICEX, 2019) Adicionalmente, se menciona el crecimiento de 12% en el valor FOB de las agroexportaciones alcanzadas en el año 2018 respecto al año anterior. Con respecto a las agroexportaciones, el Perú se encuentra entre los 10 primeros exportadores mundiales de los siguientes productos agrícolas: 7 Figura 1. Ranking de producto agrícolas peruanos exportados Fuente: (PromPerú) El auge de este sector económico genera en las empresas tener mayor demanda y por ende a incrementar su capacidad productiva, lo cual requiere de una importante inversión. En este punto de crecimiento a nuevos mercados internacionales hace que las grandes empresas agroexportadoras opten por invertir en nuevos proyectos agrícolas. Barrientos Felipa P. menciona que la participación de la agricultura en la economía peruana impulsa a la generación de divisas y empleo a través de la adición de nuevos productos agrícolas por exportar a la demanda del mercado internacional. Asimismo, la exportación de productos agrícolas tradicionales está representadas por el algodón, la caña de azúcar y el café, representando así productos agrícolas commodities internacionalmente. (Barrientos Felipa, 2018) La Agricultura en el Perú ha ido teniendo cada vez más mayor participación en el mercado internacional con producto agrícolas, que aún no cuentan con estándares de consumo y con poco valor agregado, lo cual hace que los otros países competidores no se sientan amenazados frente a sus productos. No obstante, la agricultura ha adquirido en los últimos años nuevos productos como la quinua, uva, palta, mango, banano orgánico y demás, los 8 cuales representan un crecimiento importante en las exportaciones hacia el exterior. (Barrientos Felipa, 2018) 1.1.2 Riesgo del sector Agrícola: Según el informe “Agroindustrias para el desarrollo” señala que el limitado acceso al financiamiento que tienen las empresas en este sector es uno de los problemas claves para el no desarrollo en la agroindustria. El sector Agrario frecuentemente suele tener problemas para acceder u obtener un capital de trabajo para expandirse o adquirir nuevas empresas. (FAO, 2013) Adicional a lo ya descrito, se tiene que los riesgos de los agronegocios difieren o son muy distintos al de los otros sectores económicos, debido que se enfrenta a riesgos de producción y precios anuales de intra e inter-comercialización. Es de esta manera, que se comienza a generar una especie de restricción crediticia en el sector por ignorancia de la entidad bancaria comercial sobre el sector y su potencial crecimiento a largo plazo. Por esta razón, es de suma importancia que las empresas de este sector cuenten con derechos de propiedad sobre sus plantas productoras, ya que esto les servirá de forma de aval y/o garantía para acceder a servicios financieros tradicionales. El FAO también indica lo siguiente: “Dado que las agroindustrias tienen un alto riesgo, pero son un segmento de la economía con un margen relativamente bajo, su éxito exigirá maneras flexibles e innovadoras de cobertura contra el riesgo.” De acuerdo con la información observada podemos determinar que el tema en investigación y sus variables no han sido muy investigados de manera conjunta. Por lo cual, con la presente tesis se busca presentar un tema de coyuntura y que aporte al sector económico en estudio. Asimismo, pudimos observar en artículos y revistas de otros países que el tema si ha sido investigado, pero desde otra perspectiva a la que se tiene en mente dar a la tesis en curso. No obstante, de dichos estudios académicos se utilizaron la información de manera pertinente para el desarrollo de nuestro estado de cuestión a la problemática. 9 1.1.3 Uso Fideicomiso en la Gestión Financiera Según señala Cómitre P., el fideicomiso también es conocido en el mundo empresarial como una herramienta financiera de seguridad y flexibilidad que permite a las empresas mitigar sus riesgos y acceder a un financiamiento con mejores tasas, como también a garantizar el pago de sus obligaciones financieras con la entidad financiera. (Comitre, ¿Cuáles son las ventajas de un fideicomiso?, 2014) El proceso de la creación de un fideicomiso se inicia cuando el fideicomitente cuenta con una necesidad de financiamiento para alguna inversión que necesita verse satisfecha, por lo cual como forma de garantía ofrece al banco y/o entidad que satisfaga dicha necesidad, un bien parte de su patrimonio. Este bien es denominado el Patrimonio autónomo fideicometido, el cual es una figura jurídica separada e independiente del fideicomitente, fiduciario y fideicomisario. No obstante, la entidad financiera fiduciario es quien cuenta con el demonio administrativo de dicho patrimonio, por lo que se encuentra con el deber de remitir informes periódicos sobre las fluctuaciones del valor de dicho activo. (Mac Lean M., 2009) Según (Comitre, ¿Cuáles son las ventajas de un fideicomiso?, 2014), en una entrevista al Diario Gestión, un fideicomiso al ser un contrato con características flexibles y de alta confiabilidad, generaba que el riesgo de la operación financiera en cuestión disminuya. Es decir, que el fideicomiso cumple también como un separador de riesgos lo que genera al cliente conseguir menores tasas de interés. “A menor tasa, menor riesgo; y, a mayor riesgo, mayor tasa” Como señala (Comitre, Bazan, Farfan, Gutierrez, & Valdez, 2015), el fideicomiso representa un tipo de contrato flexible entre en las partes, la empresa y el banco, la cual se puede adaptar a las necesidades que lo requiera el capital. Es decir, que en épocas de recesión para las empresas el uso de esta herramienta permite generar confianza y en épocas de crecimiento, como la de nuestro sector en estudio, ayuda a canalizar los excesos de recursos a través de una mitigación de riesgos. 10 Asimismo, (Cruz-de los Heros, 2018) menciona en su Investigación Académica “El Fideicomiso de Garantía” que el tipo de fideicomiso de mayor uso en el Perú es el de Garantía, ya que cuenta con múltiples ventajas y beneficios, siendo su mayor participación en los sectores de construcción, energía, inmobiliarios, retail, agroindustrias y entre otros. Por otro lado, (Sotomayor Gutierrez & Zarate Dias, 2019) señalan que las altas tasas de interés ofrecidas por las entidades del sistema financiero para un préstamo de alto rango influyen en la toma de decisiones de las empresas y sus proyectos. Como también mencionan que el fideicomiso se presenta como una figura contractual que puede servir para diferentes proyectos de inversión que requiera una empresa. Cabe resaltar que nuestra investigación se basará en el impacto del fideicomiso y el comparativo frente a otras herramientas de financiamiento para poder elegir la opción optima según el sector agroexportador. Este comparativo tuvo un punto de vista más extenso en la investigación (Kennedy Gimenez, 2014), especialista en temas legales cuya investigación realizada en Asunción-Paraguay en el 2014 se basó, como llamo su título, en Las Ventajas del fideicomiso de garantía respecto de las clásicas garantías reales. Citaremos este antecedente para aplicarlo en el sector peruano y sus reacciones en la economía actual sabiendo que el comparativo con las otras opciones de financiamiento como préstamos bancarios y las hipotecas también están en uso en Perú en 2018. Para el entendimiento del tratamiento contable para el uso del fideicomiso, se realizó una investigación el presente año con la finalidad de dirigir su impacto tributario para la renta. (Olano Romero, 2019), estudiante de la Pontificia Universidad Católica de Perú realizó la investigación titulada Análisis crítico del fideicomiso peruano. Entendiendo su tratamiento impositivo de renta cuyo fin era demostrar la reacción en la renta de esta herramienta financiera. 11 1.2 El Fideicomiso Comenzaremos explicando los tipos de orígenes que el termino o la figura de “Fideicomiso” tiene, a continuación, haremos una breve explicación de cada uno de ellos: a) En el derecho Romano: El término de Fideicomiso se origina de las palabras en el idioma latín “fides”, de significado ‘fe’ y “commisus”, de significado ‘confianza o confiado’. Por lo cual, se puede decir que el Fideicomiso era y es un término que connota ‘Confianza’ desde la perspectiva del derecho romano. (Comitre, Bazan, Farfan, Gutierrez, & Valdez, 2015). Según (De la Fuente-Hontañon, 2012) en el derecho Romano, el “Fideicommissum” era de origen sucesorio o testamentario. Es decir, el Testador (persona que realiza el testamento manifestando su voluntad en dicho documento) designa a un heredero como persona de confianza para que administre los bienes heredados para beneficios de una tercera persona mediante una transferencia posterior. Esto en la época Romana no iba de acuerdo con las restricciones que se tenía en ese entonces, lo cual resultaba una evasión al derecho de esa época. De la Fuente (2012) también indica en su informe que existe una probabilidad que hasta el mismo Emperador Romano de esa época haya tenido este tipo de “Testamento o Sucesión Hereditaria”, lo cual implicó la modificación de este término en el derecho Romano lo cual termino haciendo al Fideicomiso una figura del Derecho civil de esa época. Sin embargo, se encontró que la estructura y métodos de este tipo de instrumento de sucesión carecía de acciones con la finalidad de cumplir el objetivo del Testador, es decir que no se garantizaba la finalidad con la que se había constituido el Fideicomiso en un principio. 12 b) En el derecho germánico: El fideicomiso en el Derecho Germánico proviene de la influencia del Derecho Romano, no obstante, este su tuvo que ir adaptando al contexto y cultura de la sociedad germánica. En estas modificaciones se encuentra el nuevo uso del fideicomiso ya no como algo hereditario si no como “Una prenda inmobiliaria”1, es decir la entrega de un bien en garantía a favor del acreedor. Esto se encontraba amparado bajo documentos que servían de sustenta para la transferencia de la propiedad al acreedor, esto es otra diferencia que se tiene con el derecho Romano el cual se consistía únicamente en un pacto verbal entre las partes. Otro uso que le dio a la figura del fideicomiso fue a las transferencias mediante una donación a favor del acreedor, quien se reservaba un derecho de goce vitalicio sobre dicha donación o bien. En conclusión, el fideicomiso por el derecho germánico utilizo más la parte del fiduciario como el administrador del bien, quien tenía que cumplir con sus obligaciones estipuladas por el acreedor o constituyente. En caso, el fiduciario incumplía con sus obligaciones el acreedor podía implementar acciones que reivindicaban mediante documentos probatorios la titularidad del bien a su favor. Esto en el derecho romano no se podía dar, ya que la transferencia era plena y real a favor total del fiduciario. (Comitre, Bazan, Farfan, Gutierrez, & Valdez, 2015). c) En el derecho anglosajón: En este tipo de derecho, conocido común como el Derecho Ingles, se tiene que entender 2 términos, el common law y el equity, los cuales tienen diferencias específicas. Por un lado, el common law se entiende como el derecho común aplicable para todo el país, es decir es el conjunto de jurisprudencia que los abogados y jueces tienen para resolver los casos o controversias en el país. Por otro lado, el equity fue usado en su época por la clase política inglesa a través del canciller más no por los 1 Conocido como el derecho real que tiene como función el asegurar al acreedor el cumplimiento y satisfacción del crédito otorgado a su favor. 13 jueces, esta herramienta busca tratar a las partes bajo las mismas condiciones y circunstancias siendo complementaria al common law garantizando así el alcance de la justicia para todos. Con estos 2 conceptos se genera el “Trust” figura de fideicomiso utilizado en el derecho Ingles naciendo de los términos “use”, significa la transferencia de tierras a favor de una persona quien es encargada de administrarlas a favor del propietario o un tercero. No obstante, en el desarrollo de esta herramienta a lo largo de las épocas, se entendió que era una forma de burlar a las leyes y gozar de beneficios de propiedades que no les pertenecía. Por esta razón el Trust surge como excepción, incluyendo así diferencias que permiten su aplicación las leyes inglesas, tales como la Transferencias de la propiedad al beneficiario una vez cumplida la finalidad del contrato o el encargo que se designó por parte del constituyente; y, el desdoblamiento del derecho de propiedad que resulta ser el resultado de la dualidad del sistema inglés (Common law y equity), es decir la doble titularidad sobre una misma cosa. 1.2.1 Características del Fideicomiso El fideicomiso presenta características que lo diferencian de los instrumentos financieros que se encuentran en las entidades bancarias comerciales, tales como: o Es de carácter multilateral: Las partes que conforman el fideicomiso, son como mínimo 3, y todas encuentran obligadas mediante prestaciones recíprocas entre estas. o Es principal: Es un contrato independiente de cualquier otro; tiene existencia por sí solo con un fin propio. 14 o Es típico: Regulado bajo la (Ley Nº 26702, 1998) LGSF donde se regula la participación del fideicomiso en el Sistema financiero peruano. o Es oneroso: Lo conforman las prestaciones con contenido oneroso, conocidos como las obligaciones garantizadas dentro del contrato, las cuales tienen carácter recíproco entre las partes. o Es formal: La constitución del fideicomiso se efectúa y perfecciona mediante un contrato suscrito entre las partes, formalizado con un público notarial o contrato privado, según sea el caso. Asimismo, (Comitre, Bazan, Farfan, Gutierrez, & Valdez, 2015; Comitre, Bazan, Farfan, Gutierrez, & Valdez, 2015) señalan que, en el caso del Fideicomiso en el Perú, este tiene ciertas características generales dado que los fideicomisos dependen mucho del marco legal del país. A continuación. Mencionaremos las principales características según el marco legal peruano: o La parte líquida de los fondos que conformen parte del fideicomiso no esta afecta a encaje. o La SBS puede remover a un fiduciario solamente en caso de dolo o culpa grave que tenga este por una mala administración del patrimonio fideicometido. o En fideicomisos de titulación, las entidades financieras autorizadas para ejercer función de fiduciarios deben constituir sociedades titularizadoras. o Existe acciones a favor del acreedor si es que existiera algún tipo de perjuicio por el contrato de fideicomiso, esta se dará dentro de un plazo de 6 meses de publicado en el diario oficial el aviso de constitución del fideicomiso; caso contrario la 15 acción tendrá que caducarse a los 2 meses de que el acreedor haya sido notificado sobre el fideicomiso constituido. o Se puede constituir bajo un testamento, al igual que en otros países de Sudamérica. o El cargo de fiduciario constituye un requisito de validez que debe tener el contrato del fideicomiso. Se exonera el fideicomiso testamentario, que se entiendo como constituido desde que se inicia la sucesión hereditaria. Si en caso el fiduciario designado no llega a aceptar el cargo se debe renombrar a un reemplazante o sustituto, si esto se rechaza por segunda vez el fideicomiso se extingue. o El fideicomiso se puede constituir a favor de cualquier tipo de persona, Sin embargo, de existir el caso de que el fideicomisario se encuentre identificado y que el mismo intervenga en el proceso de constitución del fideicomiso, este tendrá derechos a titulo propio. Ello implicaría que el fideicomisario no pueda realizar ninguna modificación en el contrato para sus beneficios. o El fideicomitente puede resolver el fideicomiso, pero únicamente el constituido a titulo gratuito. o Aquellas disposiciones hechas por el fiduciario que traten sobre los bienes del patrimonio fideicometido son anulables y también las disposiciones que exceden o contravengan las facultades establecidas en el contrato para cada una de las partes, con excepción de las transferencias que se hubiesen realizado mediante mercados organizados (Ej. Bolsa de Valores). o Existe la posibilidad que el fideicomisario, fideicomitente y herederos de ambos puedan ceder sus derechos respecto del fideicomiso a cualquier persona que no tenga impedimentos por ley o por el contrato de constitución. 16 o Las responsabilidades por los actos, contratos y cualquier operación que se realice derivados de la finalidad del fideicomiso recaerá personalmente sobre el factor fiduciario. o La empresa fiduciaria no tiene el derecho de propiedad sobre los bienes que conforman el patrimonio fideicometido. o El agente fiduciario puede designar una comisión administradora para dicho fideicomiso, o incluso contratar a personal ad hoc, dependiendo de la finalidad del fideicomiso o de la naturaleza de los actos y contratos que deban realizarse en su función. o En una estructura de fideicomiso en garantía, el acreedor a favor de quien se constituye no podrá ser designado como fiduciario. El fideicomiso es un instrumento legal y financiero que esta especializado según la necesidad del cliente, en otros términos, es el mediador legal entre el beneficiario o fideicomisario y el fideicomitente por sus obligaciones garantizadas de este ultimo con el primero. Al tener esta herramienta un carácter formal, confiable y oneroso, permite que la operación se vuelva más transparente y confiable para todas las partes del fideicomiso. 1.2.2 Partes del Fideicomiso El fideicomiso es constituido a través de una documentación formal y onerosa denominado “Contrato de Fideicomiso” el cual ingresa a registros públicos del gobierno peruano y se requiere su publicación en el Diario Nacional “El Peruano”. Esto es requerido por las entidades reguladoras de las empresas fiduciarias y la legislación peruana. 17 Asimismo, según señala La Ley General del Sistema Financiero (LGSF), en el cual se señala las regalas y normas del mercado financiero peruano, que el fideicomiso debe estar constituido por las siguientes partes formales:  Fideicomitente: Persona natural o jurídica que encarga a un fiduciario, los bienes parte de su patrimonio para un fin en específico. Además, el único requisito es que cuente con capacidad jurídica para realizar la transferencia de los bienes o derechos que conformarán el patrimonio fideicometido.  Fideicomisario: Persona natural o jurídica en cuyo provecho se desarrolla el fideicomiso y se cumple la finalidad especifica establecida en el contrato, beneficiario del fideicomiso. Solo en el caso el caso de un fideicomiso de titulación el fideicomisario y el fiduciario pueden ser la misma persona. Además, la ley contempla la posibilidad de fideicomisarios sucesivos como también la figura de contar con más 5 fideicomisarios para un mismo contrato de fideicomiso.  Fiduciario: Instituciones financieras autorizada por la LGSF para ejercer servicios fiduciarios entre ellos están las empresas de operaciones múltiples, empresas fiduciarias, compañías de seguros y reaseguros y COFIDE. Es la persona o figura en la cual recae la responsabilidad de administración del patrimonio fideicometido. Esta administración consta de la confianza y seguridad que el mismo fiduciario le da en sus operaciones, las cuales deben estar soportadas en el contrato, estas acciones también deben contar con la diligencia y dedicación de un ordenado comerciante y leal administrador. No obstante, el fideicomiso puede contar con muchas más partes que tengan que intervenir en la operación, tales como alguna institución del gobierno, quien realiza un papel fiscalizador, otra persona jurídica o natural, quien se adjudica el pago de las obligaciones directamente, y, otras 18 intervinientes que tengan roles que cumplir con el contrato del fideicomiso y/o la operación por la cual se realiza. 1.2.3 Tipos de Fideicomiso En el sistema financiero peruano, la figura del fideicomiso es utilizada como una herramienta financiera especializada. Es decir, que no es de común uso en las entidades financieras comerciales, las cuales ofrecen productos similares un gran ejemplo es el crédito hipotecario muy utilizado en sectores económicos emergentes. No obstante, existe empresas especializadas y autorizadas por la Superintendencia de Banca, seguros y AFP comúnmente conocida como (SBS), el mercado fiduciario esta conformado por un número pequeño de fiducias, de las cuales se destacan a La Fiduciaria S.A., FIDUPERU y CORFID por su porcentaje de participación en este sector. Siendo así la empresa líder la primera de las mencionadas, de la cual se pudo extraer la información sobre la figura del “Fideicomiso” en el Perú. En la plataforma web de esta página se pudo extraer información sobre los principales tipos de fideicomisos desarrollados en el Perú a la fecha, como también una breve descripción de cada uno (LA FIDUCIARIA S.A., 2019). A continuación, se detallará y explicará las estructuras de fideicomiso desarrolladas en mercado fiduciario peruano: a) Garantía de activos y/o flujos: La finalidad de este tipo de fideicomiso es garantizar el cumplimiento de las obligaciones que la empresa (constituyente) denominado como el fideicomitente, tenga con uno o varios terceros (entidades bancarias, entre otros) denominado como el fideicomisario. Las obligaciones pueden estar conformarse por re-pagos de financiamientos, emisiones de bonos, cumplimientos de acuerdos concretos, entre otras. El patrimonio fideicometido que se ponga a disposición de garantía puede estar compuesto de los siguientes elementos:  Activos: En esta estructura se encuentra todos los tipos de activos tales como terrenos, edificaciones, inmuebles, acciones, entre otros. Según señala la 19 página de la empresa líder en fideicomisos, esta estructura es de mayor uso en el sector industrial y agrario, pues otorgan a sus activos principales como garantía pare financiamiento de largo plazo. Figura 2. Estructura fideicomiso de activos Fuente: (LA FIDUCIARIA S.A., 2019)  Flujos: Aquí se encuentran los derechos de cobros y cualquier otro tipo de flujos dinerarios provenientes de los diferentes ingresos que tenga la empresa constituyente (fideicomitente) con la finalidad de debitar en dichas cuentas corrientes las obligaciones garantizadas que se tiene por un financiamiento liberando así los remanentes a la empresa fideicomitente. Esta estructura es de mayor uso en el sector Retail, las cuales suelen dar en cesión sus cuentas de flujos por tarjetas de crédito o contratos de alquiler (Mall’s). Figura 3. Estructura fideicomiso de flujos Fuente: (LA FIDUCIARIA S.A., 2019)  Mixto: Es la combinación de los dos tipos de patrimonio fideicometidos mencionados anteriormente, cumpliendo así la misma operatividad en su estructura. Con esto se puede generar un solo fideicomiso o dos separados por 20 patrimonio. Su aplicación se da mayormente en Project Finance y en aquellas operaciones de financiamiento que tengan altos riesgos y requerían un nivel más alto de garantías. Figura 4. Estructura de fideicomiso mixto Fuente: (LA FIDUCIARIA S.A., 2019) b) Fideicomiso de Escrow – Condicionado: En esta estructura de contrato de fideicomiso se tiene como objetivo proporcionar al cliente la confianza en determinada operación de “Compraventa de Activos (Empresas)” para que así se realice con mayor transparencia y factibilidad. En este caso el patrimonio fideicometido se conforma por los activos a venderse con sus respectivos precios, los cuales deben mantenerse en el patrimonio hasta que lleguen a ciertos acuerdos mutuos por las partes. Cuando se cumplan las condiciones estipuladas, se transfieren los activos en venta al comprador y sus respectivos precios al vendedor.  Compra de empresas: En el caso que sea venta-compra de Empresas, la estructura facilita a las partes el no intercambio de acciones y flujos dinerarios solo hasta contar con un reporte final de un auditor determinado por las partes. Asimismo, con la constitución del fideicomiso se asegura que las condiciones que se pactaron entre las partes, contando una vez con el reporte del auditor, están se cumplan en su totalidad. 21  Compra de inmuebles: En el caso de Compraventa de inmuebles, la estructura indicada se adecua para fortalecer las garantías de las partes que conformen el fideicomiso en conjunto todo ello establecido en el contrato. Figura 5. Estructura de fideicomiso de Escrow – Condicionado Fuente: (LA FIDUCIARIA S.A., 2019) c) Fideicomiso de Puente Inmobiliario: La estructura de Puente Inmobiliario tiene como objetivo garantizar el uso de un terreno determinado para la realización de un proyecto inmobiliario, todo ello mientras se realizan los estudios y tramites municipales necesarios para llevarlo a flote. En este caso, el patrimonio fideicometido esta conformado por el Terreno en cuestión, adquirido para la construcción del proyecto por parte del fideicomitente (Constructor) mediante un financiamiento otorgado por un fideicomisario (Inversionista), quien participa durante todo el proyecto inmobiliario. Esta estructura de fideicomiso puede tener modificaciones para formar parte de un fideicomiso inmobiliario, lo cual facilitaría la administración de todo el proyecto en general hasta su finalización. 22 Figura 6. Estructura del fideicomiso de puente inmobiliario Fuente: (LA FIDUCIARIA S.A., 2019) d) Fideicomiso Inmobiliario: Esta estructura tiene como objetivo asegurar que el desarrollo de un proyecto inmobiliario se dé con transparencia, facilidad y orden, y realizar todos los pagos del proyecto (costos de la obra, pagos del financiamiento, entre otros). No obstante, el responsable frente a los adquirientes seguirá siendo el promotor/constructor (fideicomitente) del proyecto. El patrimonio está compuesto por todos los activos que forman parte de un proyecto inmobiliario, teniendo la posibilidad de estructurar dos fideicomisos en vez de uno. En esta alternativa, se tienen un Fideicomiso de Activos compuesto por el terreno, planos, seguros, entre otros; y un Fideicomiso de Flujos compuesto por los derechos de cobro del fideicomitente (los aportes dinerarios del promotor/constructor, pagos de los adquirientes, entre otros). En esta estructura es necesario contar con un banco "patrocinador" del proyecto y un supervisor elegido por este, quien aprueba los desembolsos dinerarios según el avance de la obra. 23 Figura 7. Estructura del fideicomiso de proyectos inmobiliarios Fuente: (LA FIDUCIARIA S.A., 2019) e) Fideicomiso Control de flujos (Carta Fianza): Bajo la estructura de Control de flujos, tenemos al termino “Carta Fianza”, esta estructura de uso común en obras públicas y privadas licitadas que tienen como objetivo administrar los fondos otorgados por la entidad que las contrate, en este caso al ganador de la licitación para la realización de una obra especifica. Cabe mencionar que, para participar de la obra en licitación, el fideicomitente (Buena Pro) tiene que haber entregado las “cartas fianza” a la entidad. De esta forma, se administra los fondos mediante un Fideicomiso, donde se minimiza en lo posible el riesgo de ejecución de las cartas fianzas y asegurar el destino de los fondos específicamente para el desarrollo de la obra. En este caso el patrimonio se encuentra compuesto por los “derechos de cobro” sobre los desembolsos de los fondos realizados, en el cual participa un determinado “Supervisor”, quien debe ser elegido por el Fideicomisario, su rol es aprobar los desembolsos solicitados para el desarrollo de la obra de acuerdo en el avance en que se encuentre la misma. 24 Figura 8. Estructura del fideicomiso control de flujos - Carta Fianza Fuente: (LA FIDUCIARIA S.A., 2019) f) Fideicomiso de Acuerdo de Accionistas La estructura de Acuerdo de accionistas tiene como objetivo administrar determinadas acciones de una organización mediante el acuerdo de accionista, asegurando así que lo pactado en dicho “Acuerdo” se cumpla. En este caso el patrimonio fideicometido estaría conformado por las acciones propias de la empresa, las cuales deben ser cedidas por el grupo de Accionistas, cuyas decisiones se encuentran reguladas por la figura del Fiduciario, dicho rol debe estar indicado en el Acuerdo de accionista. Figura 9. Estructura del fideicomiso de acuerdo de accionistas Fuente: (LA FIDUCIARIA S.A., 2019) 25 g) Fideicomiso de Patrimonio Familiar En el caso de Patrimonio familiar, su estructura es similar al de Acuerdo de accionistas debido que se administran acciones de “titularidad” de un determinado grupo familiar, esto se puede dar en una o varias empresas en beneficio del grupo familiar. Esta estructura tiene como objetivo mantener el patrimonio familiar unificado (Un solo paquete de acciones) y asegurar así a los futuros herederos de los accionistas iniciales, quienes se beneficiarían del fideicomiso sin tener que recibir las mencionadas acciones. Es decir, los herederos perciben los derechos fideicomisarios de la operación, donde no existe el retorno del patrimonio. Por ende, estos beneficiarios no cuentan con el poder de disponer sobre estas mismas acciones. El patrimonio fideicometido para este caso este compuesto por las acciones del grupo familiar y/o fondo de la empresa familiar, los cuales serán administrados por el fiduciario soportado bajo un comité donde se presenten las partes. Figura 10. Estructura de fideicomiso de patrimonio familiar Fuente: (LA FIDUCIARIA S.A., 2019) h) Fideicomiso de Gestión de Deuda En la estructura de Gestión de Deuda, a diferencia de los otros casos ya explicado en la presente investigación, en esta el fideicomitente ya no asume como lo es usualmente la deuda frente a un fideicomisario. En esta estructura el fideicomiso de Gestión es quien asume la deuda directamente, mediante una suscripción de un contrato de crédito, pagaré o leasing. De esta forma, se asegura el cumplimiento de 26 las obligaciones generadas por un determinado financiamiento, en donde el fideicomitente se aísla y se mantiene únicamente en el patrimonio autónomo generado por la gestión de deuda. En dicha operación se constituye dos fideicomisos: o Fideicomiso de Gestión: El fideicomitente cede al fideicomiso constituido, sus activos y/o derechos de cobro y flujos. Luego, el propio fideicomiso en su autonomía suscribe una el contrato crediticio con un prestamista o financiero (fideicomisario), garantizando así la obligación del crédito mediante el Fideicomiso de Garantía. o Fideicomiso de Garantía: Al fideicomiso de Gestión ceder su patrimonio (Activos, derechos de cobro y/o flujos) para constituir un nuevo fideicomiso de garantía, para así asegurar el pago de las cuotas crediticias generadas. Este tipo de fideicomiso se fueron desarrollando en proyectos de líneas de transmisión eléctrica en el Sector Energía. Al resultar exitoso para el cliente, se fue implementando en el desarrollo de centros comerciales y empresariales en el Perú. Figura 11. Estructura de fideicomiso de gestión de deuda Fuente: (LA FIDUCIARIA S.A., 2019) 27 1.2.4 Contexto del Fideicomiso en el Perú En el Perú el Fideicomiso se encuentra regulado bajo la (Ley Nº 26702, 1998) , señalando que es el acto jurídico establecido en un contrato en el cual las partes representan un acto de confianza en virtud de la entrega de un patrimonio afecto a un objetivo en específico en favor del beneficiario del fideicomiso. El fideicomiso se encuentra regulado en Perú por la Resolución SBS N° 1010-99 decretado el 11 de noviembre de 1999 donde explica las definiciones de fideicomiso, así como sus requisitos y los tipos aceptados, algunas clases especiales y las normas prudenciales. Tener en cuenta: “Que, la Ley General del Sistema Financiero y del Sistema de Seguros y Orgánica de la Superintendencia de Banca y Seguros - Ley Nº 26702, modificada por las Leyes Nº 27008 y Nº 27102, en adelante Ley General, en su Artículo 242 establece que las empresas de operaciones múltiples, empresas de servicios fiduciarios, empresas de seguros, empresas de reaseguros y COFIDE pueden desempeñarse como fiduciarios en operaciones de fideicomiso” Específicamente en Perú, el uso del fideicomiso ha sido relativamente nuevo siendo una opción poco conocida en la comunidad peruana y por ende poco investigada. En la Pontificia Universidad Católica de Perú se realizó una investigación sobre El sistema fiduciario en el Perú: propuesta para la adopción de un sistema fiduciario mixto realizado por (Vivanco Luyo, 2017), esta información nos servirá de base para entender la situación y el uso moderno del fideicomiso, así como las características modernas que tiene esta nueva opción de financiamiento para la población peruana.	11,407
https://openalex.org/W3181789323	OpenAlex	Open Science	CC-By	2,021	Centralized vs. distributed energy storage – Benefits for residential users	Behnam Zakeri	English	Spoken	11,708	19,081	Aalborg Universitet Centralized vs. distributed energy storage – Benefits for residential users Zakeri, Behnam; Gissey, Giorgio Castagneto; Dodds, Paul E.; Subkhankulova, Dina Published in: Energy DOI (link to publication from Publisher): 10.1016/j.energy.2021.121443 Creative Commons License CC BY 4.0 Publication date: 2021 Document Version Publisher's PDF, also known as Version of record Link to publication from Aalborg University Citation for published version (APA): Zakeri, B., Gissey, G. C., Dodds, P. E., & Subkhankulova, D. (2021). Centralized vs. distributed energy storage – Benefits for residential users. Energy, 236, Article 121443. https://doi.org/10.1016/j.energy.2021.121443 Centralized vs. distributed energy storage – Benefits for residential users Citation for published version (APA): Zakeri, B., Gissey, G. C., Dodds, P. E., & Subkhankulova, D. (2021). Centralized vs. distributed energy storage – Benefits for residential users. 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Take down policy If you believe that this document breaches copyright please contact us at vbn@aub.aau.dk providing details, and we will remove access to the work immediately and investigate your claim. Citation for published version (APA): Zakeri, B., Gissey, G. C., Dodds, P. E., & Subkhankulova, D. (2021). Centralized vs. distributed energy storage – Benefits for residential users. Energy, 236, Article 121443. https://doi.org/10.1016/j.energy.2021.121443 a r t i c l e i n f o Article history: Received 31 December 2020 Received in revised form 4 July 2021 Accepted 6 July 2021 Available online 10 July 2021 Article history: Received 31 December 2020 Received in revised form 4 July 2021 Accepted 6 July 2021 Available online 10 July 2021 Keywords: Electrical energy storage Energy policy Energy system model Decentralized energy Value of energy storage Smart energy systems Distributed energy storage is a solution for increasing self-consumption of variable renewable energy such as solar and wind energy at the end user site. Small-scale energy storage systems can be centrally coordinated by "aggregation" to offer different services to the grid, such as operational ﬂexibility and peak shaving. This paper shows how centralized coordination vs. distributed operation of residential electricity storage (home batteries) could affect the savings of owners. A hybrid method is applied to model the operation of solar photovoltaic (PV) and battery energy storage for a typical UK householder, linked with a whole-system power system model to account for long-term energy transitions. Based on results, electricity consumers can accumulate greater savings under centralized coordination by between 4 and 8% when operating no technology, by 3-11% with electricity storage alone, by 2-5% with stand- alone solar PV, while 0-2% with PV-battery combined. Centralized coordination of home batteries of- fers more optimized electricity prices in the system, and as such, higher private savings to all consumers. However, consumers without onsite energy technologies beneﬁt more than PV-battery owners. There- fore, based on system-level beneﬁts of aggregation, the regulator should incentivize prosumers with PV- battery, who are able to balance their electricity supply-demand even without central coordination, to let their storage be controlled centrally. Possible revenues of storage owners from ancillary services as well as the cost of aggregation (e.g., transaction fees charged by aggregators) are not considered in this analysis. Keywords: Electrical energy storage Energy policy Energy system model Decentralized energy Value of energy storage Smart energy systems © 2021 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Electricity generation from solar PV is not always correlated with electricity demand. For example, in cold climate countries electricity demand peaks typically happen in the evenings when there is no solar energy [1]. Downloaded from vbn.aau.dk on: October 23, 2024 Downloaded from vbn.aau.dk on: October 23, 2024 Energy 236 (2021) 121443 Behnam Zakeri a, b, c, d, , 1, Giorgio Castagneto Gissey b, 1, Paul E. Dodds b, Dina Subkhankulova b Behnam Zakeri a, b, c, d, , 1, Giorgio Castagneto Gissey b, 1, Paul E. Dodds Dina Subkhankulova b a Energy, Climate, and Environment Program, International Institute for Applied Systems Analysis (IIASA), Austria b UCL Energy Institute, University College London, UK a Energy, Climate, and Environment Program, International Institute for Applied Systems Analysis (IIASA), Austria b UCL Energy Institute, University College London, UK c Energy Efﬁciency and Systems, Aalto University, Espoo, Finland d Sustainable Energy Planning, Aalborg University, Copenhagen, Denmark c Energy Efﬁciency and Systems, Aalto University, Espoo, Finland d Sustainable Energy Planning, Aalborg University, Copenhagen, Denmark * Corresponding author. IIASA, Schlossplatz 1, Laxenburg, Austria. E-mail address: zakeri@iiasa.ac.at (B. Zakeri). 1 Authors with equal contribution. a r t i c l e i n f o There are different solutions for increasing the consumption of solar PV onsite, or so called “self- consumption”, which can maximize the beneﬁts of distributed energy generation and minimize the electricity bills of the PV owner [2]. One of the common solutions is to export extra elec- tricity from solar PV to the grid. However, in large-scale penetration of distributed solar PV, the export of electricity from many build- ings to the distribution grid at peak generation times will cause contingencies and grid imbalances [3], resulting in additional costs for the system [4]. Moreover, the value of self-consumption of solar electricity for the private owner is typically much higher compared to the gains from exporting electricity to the grid, as export tariffs are typically lower than purchasing electricity prices [5]. Therefore, 1.3. Private and system-level value of solar PV and energy storage 1.3. Private and system-level value of solar PV and energy storage The private value of solar PV and EES to consumers is the ﬁnancial gain that a consumer can obtain by reducing its electricity bills [30]. Wholesale electricity prices vary widely on an hourly or half-hourly basis and are typically the largest component of elec- tricity costs of consumers, comprising nearly 40e60% of their electricity bills in Europe [20]. Most prosumers4 have been early adopters, environmental enthusiasts, looking for energy security by being independent from the grid, and/or motivated by social and peer effects; not necessarily motivated purely by cost-beneﬁt analysis [31,32]. Yet the savings that prosumers with EES could achieve is a key indicator to show if more widespread adoption of such distributed energy technologies is likely to occur in the future or not. However, the cost of batteries are still at the start of their learning curves [14], which diminishes the ﬁnancial viability of investment in such technologies, from a private owner's perspec- tive [2]. Different studies show that a PV-EES system is not economically viable under current market conditions in different countries without additional ﬁnancial supports [15]or policy in- centives [16,17]. These policies are, for example, capital subsidies [8], enhanced time-of-use tariffs [18,19], peer to peer trading [20], or provision of revenue stacking3 [21]. Revenue stacking is consid- ered as one of the most effective support mechanisms for enhancing economic proﬁtability of EES systems [22], which can be possible by combining the onsite use of EES with offering grid services, such as balancing the load and/or ancillary services as shown in Refs. [23,24]. Numerous studies have investigated the proﬁtability of con- sumer investments in solar PV and EES. Many studies have derived the cost of electricity and assessed the proﬁtability of investments by considering metrics such as the Net Present Value (NPV), In- ternal Rate of Return (IRR), or the Return on Investment (ROI) of the investment. Other work adopts the “grid parity” concept to eval- uate the proﬁtability of storage by considering the levelized cost of electricity [33]. These studies, however, do not take a whole elec- tricity system approach for modelling the future electricity prices, on which the economic proﬁtability of PV-storage systems de- pends. 1.1. Distributed solar PV and energy storage 1.1. Distributed solar PV and energy storage Many governments worldwide plan to increase the share of renewable energy for environmental, economic, and energy secu- rity reasons. For achieving renewable energy targets, different in- centives and support schemes have been put in place to promote the deployment of renewable energy through decentralized and distributed generation, e.g., through solar photovoltaic (PV) at consumer sites. B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 Energy 236 (2021) 121443 the private owner of solar PV prefers to ﬁnd different ways to in- crease their self-consumption, e.g., by storing electricity via elec- trical energy storage2 (EES) systems such as batteries [6]. the private owner of solar PV prefers to ﬁnd different ways to in- crease their self-consumption, e.g., by storing electricity via elec- trical energy storage2 (EES) systems such as batteries [6]. Aggregators can offer the combined capacity of EES technologies in wholesale electricity markets, to meet the needs of the System Operator for load management and ancillary services, e.g., for Fast Frequency Response (FFR) [26]. Different studies have shown that the aggregation of small-scale EES systems could reduce the risk of higher electricity prices at peak times [27], improve social welfare [28], and increase the integration of renewable energy in the grid [29], compared to uncoordinated, independent management of such assests by their owners. As consumers are unlikely to be able to provide such services and exploit arbitrage beneﬁts simulta- neously, they may operate their resources in a way that minimizes their own electricity bills, irrespective of the potential system-level beneﬁts they could offer through aggregation [27]. Fig. 1 illustrates the main features of these two schemes for the operation of distributed energy storage, i.e., the uncoordinated operation of EES by multiple owners for their private beneﬁts (a), versus a centrally coordinated operation of small EES systems through an aggregator. EES can balance the mismatch between onsite solar PV gener- ation and electricity demand by storing electric energy at hours of low demand in daytime and discharging that to meet evening peaks. Different studies have shown that pairing solar PV with batteries (PV-EES) increases self-consumption of solar energy onsite [7] and can offer signiﬁcant cost savings to the private owner. For example, Zhang et al. 3 Revenue stacking or aggregation of beneﬁts means using an EES device for offering multiple services, such as energy arbitrage, balancing services, and T&D support; and receiving revenues for each service. 2 The terms EES, “electricity storage”, “energy storage”, and “storage” are inter- changeably used in this paper for referring to technologies that can store electricity and discharge it back at a reasonable response time. Examples of such technologies include secondary electro-chemical batteries, ﬂow batteries, pumped hydropower storage (PHS), etc. 4 Prosumers are deﬁned as consumers with the ability to produce electricity from solar PV. 1.3. Private and system-level value of solar PV and energy storage A recent study considers the impacts of a changing elec- tricity system on the consumer savings, but does not account for potential impacts of the development of demand-side technologies on the system [2]. This paper extends the previous work by ac- counting for the impact of the EES on the system, which, if neglected, may overestimate the potential beneﬁts of the EES for the owner. Because the larger the capacity of EES in the system offered by many private owners, the lower the value of arbitrage for each EES owner as the price gap between peak and off-peak will diminish. 1.1. Distributed solar PV and energy storage [8] shows that paring solar PV with a home battery in California and Hawaii is a feasible investment with a payback period of less than 10 years for different building types, while others demonstrate possible cost savings for PV-battery owners in high latitude countries in Europe under different en- ergy storage policies [9]. Also, from the system operator's perspective, distributed EES devices can contribute toward balancing the (distribution) grid by reducing peak contingencies [10] and grid management costs [11]. This can offer the Trans- mission and Distribution (T&D) grid operator signiﬁcant cost sav- ings for postponing T&D investments and grid fortiﬁcation measures at the low-voltage level [12,13]. 1.2. Coordination of distributed solar PV-storage systems Last but not the least, Ahmadi et al. [42] applies a two-stage optimal coordination of central and local EES for showing the impact on system cost reduction and voltage proﬁle enhancement. social value. The social (system) value of these resources will depend on whether these resources are being operated to reduce electricity system costs, a beneﬁt for all consumers, or to minimize private electricity costs. Solar PV may reduce electricity demand if it is subject to individual coordination by cost minimizing consumers, which would reduce prices for all consumers in the system [27]. Privately coordinated EES could increase electricity prices as there is potential that most of EES owners charge simultaneously at low price hours resulting in signiﬁcant increase of electricity demand and prices in those hours, affecting all electricity consumers. But private EES devices could also reduce peak demand, hence prices, if they were optimally operated in coordination, lowering electricity prices for all consumers [34]. However, none of the reviewed studies investigate the impact of the aggregation of distributed energy technologies (here PV-EES) on the private value of such technologies, i.e., the additional cost or beneﬁt that the owner bears for letting the aggregator coordi- nate their PV-EES. This is an important question as the deployment of EES by consumers might be affected by the way the technology is operated throughout the system. Answering this question could reduce the uncertainty consumers face when investing in battery storage, thereby facilitating further deployment of storage re- sources when needed. This would help the electricity system to reduce costs and improve security of supply by making such re- sources available to provide multiple other system services. In this respect, it is crucial to understand how the deployment of EES re- sources by consumers could be affected as more EES is aggregated throughout the electricity system. Our study investigates this too. Several studies focusing on EES in different countries have concluded that centralized coordination of distributed energy re- sources could offer numerous system-level advantages. For example, central coordination of EES can offer required ﬂexibility in matching load and supply, reducing the cost of procuring ﬂexible capacity for the system [35,36]. The value of aggregation to an electricity system has been shown to increase as more consumers are aggregated [37], with small contributions by each customer leading to large reductions in electricity costs for all consumers [38]. 1.2. Coordination of distributed solar PV-storage systems Providing grid services in many power systems is regulated by the System Operator with some technical requirements for candi- date technologies. These requirements are commonly speciﬁed as response time, availability, reliability, minimum capacity rating, etc. For example, the requirement for an energy technology for providing balancing services in Finland is a minimum power output of 5 MW [25]. These requirements leave many distributed tech- nologies such as PV-EES systems with a typical size of a few Kilo- watts unqualiﬁed for entering such marketplaces. To overcome such barriers of entry, the available capacity of many small-scale distributed technologies can be aggregated and coordinated by aggregators, which are typically third-party companies beneﬁting from control and transaction fees. Therefore, the owner of a PV-EES system can operate their asset either independently mainly for managing their own generation and demand or, alternatively, they could offer their available storage capacity to be coordinated with other small-scale EES units to participate in wholesale electricity markets through aggregators. The value of solar PV-EES to consumers is different from the value they may offer to the wider electricity system. Solar PV-EES and other distributed energy technologies could provide the elec- tricity system with different services, while offering energy security and cost savings to the owner. However, maximizing the private value of distributed technology may not simultaneously offer the highest system-wide value. Energy security has a private value to the consumer, whereas the ﬂexibility it offers to the system has a 2 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 Fig. 1. Schematic representation of uncoordinated (a) and centrally coordinated (b) operation of distributed electricity storage devices. The main characteristics of each mode of operation, including beneﬁts for the system and the private owner is depicted under each scheme. Fig. 1. Schematic representation of uncoordinated (a) and centrally coordinated (b) operation of distributed electricity storage devices. The main characteristics of each mode of operation, including beneﬁts for the system and the private owner is depicted under each scheme. Fig. 1. Schematic representation of uncoordinated (a) and centrally coordinated (b) operation of distributed electricity storage devi operation, including beneﬁts for the system and the private owner is depicted under each scheme. [41], a whole-system comparison of centralized versus decentral- ized electricity planning is carried out, showing that coordinated planning can save between 7% and 37% of the total system costs. 1.2. Coordination of distributed solar PV-storage systems It is also argued that distributed energy devices could improve social welfare under efﬁcient aggregation and coordinated opera- tion of technologies [28]. Castagneto Gissey et al. [27] investigated the impact of centralized and distributed scheduling of EES on electricity prices, highlighting that a centralized coordination offers 7% lower mean electricity price and 60% lower price volatility in the system. Sousa et al. [39] compares a peer-to-peer (P2P) versus a community market for energy trade, concluding that P2P trade offers the highest social welfare. It is further shown that the aggregator can control the capacity of distributed EES to manage the frequency deviations in the grid in a more effective way [40]; another system-level beneﬁt for all consumers. In a recent study 1.4. Objectives of this study As mentioned earlier, pairing solar PV with EES can maximize the self-consumption of PV electricity for consumers who adopt the technology and minimize their electricity costs. Yet it remains un- clear how the savings that these consumers can expect from their storage device might be affected by the way of coordination of EES in the electricity system. This paper investigates how aggregator- led and consumer-led operation of EES capacity might affect the private economic value of solar PV and EES for a UK electricity consumer with typical domestic electricity consumption. Different future developments of the energy system are explored to analyse 3 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 capital cost of different power plants, are based on the output of the UKTM energy system model [45]. the economic savings a consumer can achieve from investing in PV and batteries. Finally, it is shown that how these savings will be affected when more EES capacity is integrated into the electricity system through aggregation. By identifying these three gaps in the literature, this paper aims to answer the following research questions: the economic savings a consumer can achieve from investing in PV and batteries. Finally, it is shown that how these savings will be affected when more EES capacity is integrated into the electricity system through aggregation. By identifying these three gaps in the literature, this paper aims to answer the following research questions: The modelling framework including the linkage between different models and modules to derive consumer savings is illus- trated in Fig. 2. This framework has been previously applied to calculate solar PV-battery consumer investments [2] and value of storage aggregation to the system and electricity prices [27]. This is extended in this study by iterating electricity demand of pro- sumers, which itself is based on the optimal scheduling of PV-EES according to retail prices, back to the electricity dispatch model. With the updated electricity demand, the electricity dispatch model generates a new set of hourly electricity prices, which will affect the retail price for all consumers, both with and without onsite energy technologies. This process, highlighted in red in Fig. 2, continues until electricity prices converge in two consecutive iterations. 1. How would aggregator-led and consumer-led operations of EES in the electricity system affect savings to a typical consumer who pairs solar PV with storage? 2. 1.4. Objectives of this study Which system evolutions or energy pathways are likely to explain the process by which EES aggregation could affect sav- ings to a consumer pairing solar PV with storage? 3. What is the relationship between savings from pairing solar PV with storage to a private electricity consumer and the level of electricity system-wide storage aggregation? In other words, how would additional aggregation of EES affect the savings to a typical consumer pairing solar PV with storage? The ESMA model has been validated on an hourly basis against both the historical data and future energy scenario developed by National Grid. The results of validation suggest that the hourly demand curve modelled by ESMA stays within an acceptable level of agreement with historical data, e.g., with an average correlations coefﬁcient of 0.92 for 8760 hourly demand data points for the reference year 2015. Similarly, the analysis of hourly electricity prices simulated by ESMA in different season shows a high degree of agreement with historical spot prices, with an average correla- tion of 0.83 in winter, while 0.91e0.93 correlation on other seasons. The comparison of the ESMA's future scenarios with those modelled by National Grid shows a very high degree of agreement, yet some slight differences exist due to different modelling as- sumptions and limitations of ESMA. A detailed analysis on valida- tion of the model is represented in Chapter 5 in Ref. [46]. The remainder of this paper is structured as follows. Section 2 provides the methodology and describes the data used in this study. Section 3 reports our main results, which are discussed in Section 4. Conclusions are drawn in Section 5. 2.1. A multi-level modelling framework 2.1. A multi-level modelling framework The modelling approach is based on soft-linking a national- level, electricity system management model (ESMA) to a con- sumer cost optimization model. The input data of ESMA, i.e., elec- tricity demand, power capacity mix, and fuel prices are based on the UK “future energy scenarios” developed by the national energy regulatory, National Grid [44]. The electricity system model ESMA is designed for evaluating the operation and dispatch of a given power system mix for a time-period of one year (8760 h). It is ideally suited to generate wholesale prices under different sce- narios for EES and the rest of the system. Wholesale electricity prices are then converted into retail electricity tariffs based on different tariff designs, i.e., time of use (ToU), static, and dynamic tariffs. These tariffs are fed into an electricity private cost minimi- zation model that optimizes the use of solar PV and EES for a consumer with a typical electricity consumption proﬁle. This framework accounts for possible future evolutions of the energy system considering how EES deployments are likely to affect sav- ings of consumers. The electricity generation costs, e.g., future Retail electricity prices are calculated by adding a time- dependent mark-up over the wholesale prices, which is assumed to account for the electricity network management and distribu- tion fees [47] (see Appendix F for calculation of prices). Static and dynamic ToU electricity tariffs are calculated based on retail prices, calibrated to historical tariff data (assuming same ratios between tariffs and retail prices as today for future years). 2. Methods Onsite, small-scale batteries and electric vehicle-to-grid storage are some examples of distributed EES technologies for private consumers. The ever-growing electriﬁcation of transport, heating and other sectors are expected to change the pattern and magni- tude of electricity demand over the coming decades [43]. Accurate modelling of electricity demand over such extended periods, i.e., 20e30 years, is crucial to understand how consumer electricity prices will vary in the future and how investment in distributed technologies will return economically. Also, transitions in the electricity supply side will affect wholesale electricity prices. Higher shares of wind and nuclear capacity in the power system will offer different electricity prices and price volatility compared to a thermal power system relying on coal and gas. Hence, assessing the ﬁnancial feasibility of investment in distributed energy tech- nologies with 20e30 years of lifetime needs to be informed by a quantitative model of the overarching energy system for repre- senting the increase in the use of non-conventional energy re- sources and possible transitions in the energy system. The applied modelling work has some limitations and short- comings. Assuming ﬁxed, average fuel prices throughout each year, i.e., ﬁxed gas or biomass prices, may not conform with reality where fuel prices change by season. ESMA does not include electricity consumers under the Economy 7 tariff who beneﬁt from a lower night tariff, which may result in a slight demand and price differ- ence in winter. ESMA represents each technology as a large power plant which is different from the strategy that each single power plant may adopt. The model is run over a 26-year period, 2015e2040, initially with the objective to optimize the consumer's utility based on the lifetimes of distributed PV-EES systems. ESMA minimizes elec- tricity costs and calculates wholesale electricity prices under the assumption of centralized and distributed coordination of demand- side EES technologies. Additional information on the modelling framework and formulation is provided in the Supporting Infor- mation (Appendix A-C). Appendix H summarizes main data sources and assumptions of the model. 2.2. Future energy scenarios The evolution of the energy system over time will impact wholesale electricity prices, and hence, consumer retail prices. A whole systems approach is adopted to account for these future transitions systematically and consistent with the National Grid scenarios, which are based on a broad stakeholder engagement and modelling. Four possible evolutions of the energy system are 4 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 Fig. 2. Relationship between different models used in this analysis. Fig. 2. Relationship between different models used in this analysis. considered according to National Grid's Future Energy Scenarios [44]. These scenarios are chosen as the basis of our analysis as they cover a wide range of future energy pathways represented across two axes for green ambition and prosperity. The GB Ofﬁce of Gas and Electricity Markets (Ofgem), the National Regulatory Authority, has reviewed these scenarios, which gives them more merit for our analysis. The ﬁnancial viability of different combinations of investments in solar PV and EES for a typical UK domestic electricity user5 are examined under different energy scenarios. The household's elec- tricity bill is dependent on the consumer's load proﬁle, and on the electricity generated from solar PV, which exhibit intra-day, monthly and seasonal variations. End users with onsite generation from PV are entitled for feed- in tariffs (FiTs) of £0.049 kWh1 for electricity generation [47] and an export-to-grid tariff of £0.043 kWh1. FiT payments are assumed to cease after 20 years and to increase with the retail price index (RPI) of 3.4% p.a [48]. An average retail electricity tariff is considered based on UK National Statistics: a static tariff of £0.15 kWh1 and dynamic ToU tariff including on-peak £0.16 kWh1 during the day (7:00e23:59) and off-peak tariff of £0.07 kWh1 at nights (0:00e6:59) [48]. Future developments of static tariffs are esti- mated based on the average of wholesale electricity price in each season. We use the static tariff as the basis to derive future values for day and night ToU tariffs (see Appendix H for more details). 5 This user is represented by a three-bedroom dwelling with a load proﬁle dis- playing mean percentage night consumption of 30% and 55% under static and Economy7 ToU tariffs, respectively [47]. 2.2. Future energy scenarios y These four energy transition pathways include: (i) Gone Green, which is the most ambitious renewable expansion scenario, where the UK meets its renewable targets; (ii) Consumer Power, a consumer-centred scenario with energy security and costs as main drivers of decisions; (iii) Slow Progression, a scenario with low ambitions for decarbonization; and (iv) No Progression, where the status quo persists and there is a negligible deployment of re- newables and EES. Gone Green has the highest ambition on re- newables and storage capacity, while No Progression is similar to the present-day energy system and has the lowest capacity of re- newables from all four scenarios. Table A3 in Supplementary Ma- terial shows the key developments of the power sector in 2030 under these future scenarios. Fig. 3 portrays the installed power capacities for each of the future energy scenarios. More details of the share of each generation mode are provided in Appendix H, Table A4. The objective of a residential PV, EES, or PV-EES owner is to minimize the private costs of electricity bills. Under ToU tariffs, the lower rate during the off-peak period is suitable for charging the storage system. When the consumer operates PV, a 4-kW PV sys- tem is considered; and for EES, a 6.4 kWhe3.3 kW battery, with a lifetime of 13 years or 5000 cycles (Li-ion batteries) [49]. The bat- tery capacity degradation and efﬁciency losses are taken into ac- count as described in Appendix B. A discount rate of 5% p.a. is assumed, based on the recommendations of the UK Committee on Climate Change. Appendix G reports the details on the consumer PV-EES optimization model and the data used for modelling PV-EES technologies. 2.3. Consumer electricity cost optimization Two cases of EES scheduling are examined, in which consumers respond to either distributed or centralized coordination. Under the former, demand-side storage resources are autonomously opti- mized by consumers. In a centralized scheduling system, an aggregator coordinates electricity dispatch from EES by iterative negotiation with consumers, whose resources it does not know, enabling them to participate in the wholesale market. Centralized coordination mimics the current arrangements for large-scale EES technologies in the UK and major worldwide liberalized markets, such as for PHS. Transaction costs relating to aggregation are neglected for simplicity. Distributed coordination reﬂects the behaviour of consumers who individually schedule their ﬂexible resources to smoothen their own demand proﬁles and minimize their own electricity bills. More information on our coordination algorithms is provided in Appendix E. The electricity costs are calculated for four consumer technology combinations: (i) no technology; (ii) an EES system (EES-only); (iii) a solar PV system (PV-only); and (iv) both a solar PV and an EES system (PV-EES). We show the value of EES, which is derived by comparing annual electricity costs in the PV-EES scenario relative to the PV-only scenario. The base case scenario for deriving the relative savings of other scenarios is the no-technology case with 5 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 Fig. 3. Electricity generation mix in each future energy scenario [44]. Fig. 3. Electricity generation mix in each future energy scenario [44 static electricity tariffs. electricity bill are compared for consumers whose EES capacity in the electricity system is coordinated by an aggregator and sched- uled centrally. The results are illustrated for four different tech- nology options under static and ToU tariffs and for each future energy scenario. 3. Results Two types of energy storage coordination, i.e., coordinated and distributed, are considered for calculations. The results are based on the data of annual electricity costs and savings, averaged over the modelling period of 2015e2040. The results are reported relative to a base case scenario, i.e., the No Progression scenario under static tariff and with no onsite energy technology investments. The results show that the consumer savings is dependent on the future energy scenarios for the entire energy system. Consumer Power scenario, in which future policies are consumer-centred and promoting distributed generation offers the highest savings for all technology combinations. Gone Green and Consumer Power sce- narios offer 18% and 22% annual savings, respectively, even in the case when the consumer has no investment in distributed tech- nologies, i.e., “No technology”. This is due to higher renewable energy in these scenarios, larger share of electricity storage, and lower electricity prices compared to No Progression. The results show that the evolution of the energy system and the scheduling coordination regime have meaningful impacts on annual savings by the consumer. Distributed coordination generally induces 4e11% lower savings than centralized coordination, whereas the system's evolution accounts for changes in savings by 1e27%. The largest savings occur in scenarios with high storage and renewable capacity. The impact of additional storage capacity in the electricity system on the savings to the consumer when aggregated to participate in the wholesale market is explored too. Fig. 4 compares the average annual savings in the electricity bill in the centralized coordination for two different types of tariffs. The results show that PV-battery offers the highest savings for con- sumers ranging between 81 and 86% depending on the future scenario. However, battery alone offers no higher beneﬁts compared to the no-technology case, as under the static tariff there will be no potential for price arbitrage by EES, as electricity prices are constant for the consumer. The annual savings of the consumer from 3.1. Private savings under centralized and distributed coordination The results of the centralized coordination is presented in Table 1. The annual electricity bills and potential savings in the Table 1 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under centralized coordination. a The savings are shown as difference relative to the base scenario, i.e., consumers having “No technology” onsite, static tariffs, a Progression). relative to the base scenario, i.e., consumers having “No technology” onsite, static tariffs, and under the business-as-usual scenario (No Table 1 A l Table 1 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under centralized coordination. Centralized coordination No Progression Slow Progression Consumer Power Gone Green Tariff Technology Bill (£ p.a.) Savingsa (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) Static No technology 574 e 541 33 449 125 470 104 EES 574 0 541 33 449 125 470 104 PV 363 211 342 232 284 290 297 277 PV-EES 107 467 98 476 78 496 82 492 ToU No technology 540 34 515 59 420 154 449 125 EES 405 169 389 185 321 253 339 235 PV 307 267 298 276 244 330 260 314 PV-EES 92 482 87 487 68 506 73 501 a The savings are shown as difference relative to the base scenario, i.e., consumers having “No technology” onsite, static tariffs, and under the business-as-usual scenario (No Progression) Table 1 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under centralized coordination. 6 Energy 236 (2021) 121443 B. Zakeri, G.C. Gissey, P.E. Dodds et al. investing in solar PV alone (without EES) varies between 37% and Fig. 4. Annual electricity bill savings for a typical consumer with different distributed energy technology options in centralized coordination, under (A) a static and (B) Time- of-use (ToU) tariff, and for different future energy scenarios. The values are the average of 2016e2040 and show % change in savings relative to the base case, which is “No- technology” under a static tariff, and No Progression Scenario with the annual cost of 574£. , y, gains substantially larger annual savings compared with the decentralized scheduling. This is valid for all combinations of technologies, tariffs and future energy scenarios. The consumer is able to accumulate greater savings in the centralized case by be- tween 4 and 8% when operating no technology, by 3e11% with EES alone, by 2e5% with PV alone, and by 0e2% with both PV and EES. More notably, the higher savings in the centralized coordination compared to the distributed scheme decline as the consumer operates more onsite technologies. Operating more technologies implies greater electricity self-sufﬁciency, hence, a lower exposure to the risk of changes in retail electricity prices, which itself is affected by the type of scheduling coordination of EES by other consumers in the system (see Fig. 5). Table 1 A l Consumers with “No technology” make higher electricity bill savings in the centralized coordination scheme due to the system operator being able to improve the balancing of load and ﬂexibility resources, which results in lower peak electricity prices in the system. The lower wholesale electricity prices beneﬁt all con- sumers, including those without investment in any distributed technology. Distributed storage scheduling results in substantially lower integration of EES capacity in the electricity supply. Through arbitrage, storage minimizes the differential between on- and off- peak prices, thereby reducing electricity system costs. Less aggre- gated storage capacity implies a lower ability for the system oper- ator to reduce electricity prices. Hence, in all scenarios, greater private electricity costs and lower private savings are observed relative to centralized scheduling. Table 2 summarizes the ﬁndings for the distributed scheduling. 3.1.1. Consumer's choice of technology and electricity tariffs 3.1.1. Consumer s choice of technology and electricity tariffs The lowest electricity cost in the no-technology case occurs under centralized coordination, Consumer Power and ToU tariffs (£420 p.a.), while the highest costs occur under distributed scheduling, Slow Progression, and static tariffs (£569 p.a.). With ToU tariffs, the EES system can provide 2e3% greater savings rela- tive to static tariffs under distributed coordination compared with centralized coordination. Under ToU, the savings in the EES-only case are £99e126 under centralized coordination versus £101e140 under distributed coordination compared to “No tech- nology” in the respective future scenario. This shows approxi- mately 7% larger savings in distributed scheduling. As the distributed coordination scenario implies a less smoothened sys- tem demand, this leaves a greater ability for the consumer to take advantage between peak and off-peak price differentials. Fig. 4. Annual electricity bill savings for a typical consumer with different distributed energy technology options in centralized coordination, under (A) a static and (B) Time- of-use (ToU) tariff, and for different future energy scenarios. The values are the average of 2016e2040 and show % change in savings relative to the base case, which is “No- technology” under a static tariff, and No Progression Scenario with the annual cost of 574£. The largest savings recorded in the EES-only case occurs under centralized coordination, Consumer Power, and ToU tariffs (£321 p.a.). Conversely, the lowest savings arise under distributed coor- dination, Slow Progression, and static tariffs (£569 p.a.). Table 2 A l Table 2 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under distributed scheduling. The savings are relative to the base case: No technology, static tariff, and No Progression scenario. Distributed scheduling No Progression Slow Progression Consumer Power Gone Green Tariff Technology Bill (£ p.a.) Savings (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) Bill (£ p.a.) Savings (£ p.a.) A. Static No technology 588 0 569 19 476 112 516 72 EES 588 0 569 19 476 112 516 72 PV 378 210 359 229 301 287 327 261 PV-EES 116 472 103 485 83 505 91 497 B. ToU No technology 559 29 541 47 442 146 491 97 EES 419 169 406 182 341 247 370 218 PV 321 267 309 279 256 332 281 307 PV-EES 101 487 91 497 71 517 80 508 Table 2 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under distributed scheduling. The savings are relative to the base case: No technology, static tariff, and No Progression scenario. Fig. 5. Centralized coordination versus distributed scheduling of consumers' energy technologies under time-of-use the (ToU) electricity tariff. The values show the % savings of centralized coordination minus that of distributed scheduling relative to the base case (hence, positive values show that centralized coordination offers greater savings). level of variable renewable generation capacity in the electricity system while inversely related to level of ﬂexible supply capacity. Savings to the consumer under centralized coordination are double in Gone Green relative to Slow Progression due to the higher var- iable renewable generation in the former case, which requires an aggregated storage for balancing variations. Table 3 reports the ratio of variable renewable capacity to each unit of ﬂexible generation capacity, as well as the change in the consumer's annual electricity costs (%) resulting from storage ag- gregation in the electricity system. There is a positive relationship between the share of variable renewables in the system, and the change in electricity prices due to centralized coordination. By dividing the latter by the former, a relatively constant relationship is observed, between 3 and 4%. Demand-side ﬂexibility will be most valuable when supply is inﬂexible, leading to greater savings in the consumer's annual electricity cost under a more system-efﬁcient coordination of storage resources. 3.3. Impact of additional storage deployments on private savings Fig. 6 shows how additional electricity storage capacity is likely to affect savings from storage to a consumer with EES. In this speciﬁc analysis, we consider ToU tariffs only as they are shown to maximize the savings that storage can provide to consumers with solar PV. Additional (aggregated) storage capacity operating in the electricity system can decrease the differential between on- and off-peak electricity wholesale prices, which could in turn reduce the retail tariff on- and off-peak differential. Consumer Power and ToU tariffs (£68 p.a.), whereas the lowest savings occur when scheduling occurs on a distributed basis, under Slow Progression and static tariffs (£103 p.a.). Overall, for different technology mixes, a distributed coordina- tion of energy storage in the electricity system, as well as Slow Progression, and static tariffs tend to minimize annual savings by the consumer. Conversely, central energy storage coordination, Consumer Power and ToU tariffs maximize savings. There is a quasi-exponential fall in the private savings as more electricity storage is installed and aggregated in the wider elec- tricity system. An increase in aggregated storage capacity from 3 GW to 17 GW implies a 20% reduction in the private annual cost savings from storage to the consumer. These results do not hold if considering distributed coordination, as non-aggregated storage capacity has no effect on the marginal savings from private storage capacity. 3.2. Future energy scenarios The results suggest that the centralized coordination of EES resources in the electricity system is always lead to greater savings (up to 11%) for a typical consumer, irrespective of the future evo- lution of the energy system. Yet the order of magnitude by which savings under centralized coordination are larger depends on the relationship between variable renewable energy capacity e mostly includes wind and PV generation e and ﬂexible supply capacity, such as gas plants. If resources are mostly centrally coordinated, consumers can reduce annual electricity costs by 8e11% in Gone Green, by 4e5% in Slow Progression, and by 4e6% in Consumer Power, relative to distributed coordination. Table 2 A l Yet the change in the elec- tricity cost from coordination is small relative to the ratio between renewables and ﬂexible supply. Fig. 5. Centralized coordination versus distributed scheduling of consumers' energy technologies under time-of-use the (ToU) electricity tariff. The values show the % savings of centralized coordination minus that of distributed scheduling relative to the base case (hence, positive values show that centralized coordination offers greater savings). Table 1 A l investing in solar PV alone (without EES) varies between 37% and 51% of the base case costs, with the lower range for No Progression scenario and the highest savings for Consumer Power. If the consumer operates solar PV without EES (PV-only), the electricity bill will decline by 37e57% compared to the no- technology case, and by between 13 and 37% relative to EES-only. The lowest electricity costs for PV-only relate to centralized scheduling, Consumer Power and ToU tariffs (£244 p.a.), whereas the largest costs arise under distributed scheduling, Slow Pro- gression, and static tariffs (£359 p.a.). The results for the battery-alone case show signiﬁcant higher savings under a ToU tariff. When the consumer electricity prices differ between off-peak and peak hours, battery can offer electricity cost savings between 29 and 41%, depending on the future energy scenario. Moreover, investing on solar PV under the ToU tariff im- proves the annual cost savings by 6e10%-point compared to the static tariff (~56£ per year). A PV-battery system offers the highest savings under ToU as well, with a slight improvement compared to the static tariff (i.e., 1-3%-point). Also, the results show that the beneﬁts of the PV-battery options are the least sensitive technology investment to future energy scenarios, offering savings ranging between 84% and 88% for the four energy scenarios. Table 2 sum- marizes the results of centralized coordination for different tariffs, technology choices, and the future scenarios. The combination of solar PV with EES implies a reduction in annual electricity costs by 81e88%, or by £476e506 annually. Therefore, the consumer reduces electricity costs by at a substantial rate of 60% compared to the PV-only case (£176e256 further sav- ings annually). On average across the future energy system sce- narios, ToU tariffs imply 12% larger savings relative to static tariffs for the consumer. In this case, annual electricity costs are between £68e73 p.a. and £71e80 p.a. in the centralized and distributed cases, respectively. When operating a PV-EES system, the consumer achieves maximum savings under centralized coordination, Under centralized scheduling of the consumer's energy tech- nologies in the electricity system, the typical electricity consumer 7 Energy 236 (2021) 121443 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Table 2 Annual electricity bills and possible savings (£ p.a.) for a typical consumer under distributed scheduling. The savings are relative to the base case: No technology, static tariff, and No Progression scenario. Table 3 able to ﬂexible supply capacity (excluding storage), and relationship with savings from demand coordination. Ratio of variable renewable to ﬂexible supply capacity (excluding storage), and relationship with savings from demand coordination. Future energy scenario Ratio of renewable energy capacity to ﬂexible supply capacity Change in annual electricity costs under central coordination (% p.a.) Gone Green 2.62 8.8% Consumer Power 1.97 5.3% Slow Progression 1.81 4.5% Fig. 6. Savings to the typical consumer due to their electricity storage relative to the installed electricity storage capacity in the electricity system. This analysis considers the centralized case with ToU tariffs. Fig. 6. Savings to the typical consumer due to their electricity storage relative to the installed electricity storage capacity in the electricity system. This analysis considers the centralized case with ToU tariffs. 4.2. Potential impact of system variables on the consumer savings to centralized coordination. These results are shown to hold true for different types of technologies and evolutions of the energy system. ial impact of system variables on the consumer saving EES could provide numerous services to the electricity system [54,55], and the possibility for storage capacity to be aggregated can reduce the cost of electricity systems by decreasing peak demand and the need for expensive peaking plants. A few studies have shown the value of storage in high-renewable, inﬂexible power systems [12,34,56]. Studies considering the role of storage in the electricity system generally do not make a distinction between private and system beneﬁts from EES, which we instead consider by considering the impacts of distributed and centralized coordination. Our ﬁndings conﬁrm those of [28,50], and [35] who reported that social welfare increases if storage resources are centrally scheduled. Similarly, Castagneto Gissey et al. [27] compared centralized and distributed coordination and suggested that con- sumers could be nudged into giving away control of their storage devices to provide system beneﬁts. They found that aggregation of EES has a lower electricity system cost compared to private oper- ation by consumers. Our study enhances this work by considering how the private savings that consumers can expect from investing in storage could be affected by the way other consumers operate their storage devices. Our work suggests that storage will be more valuable to energy storers if variable renewable capacity is on average larger than the capacity of ﬂexible supply resources such as gas power plants in the power system. 4.1. Private savings from storage and control scheme This paper shows that the savings that a typical UK electricity consumer can achieve from their EES device could increase if most consumers in the electricity system allow an aggregator to coor- dinate their storage resources. When consumers’ storage capacity is operated to minimize the private costs of these consumers, herding behaviour occurs, leading to charging the consumer EES devices at the same time of the day leading to higher electricity prices relative The impact of centralized coordination of storage resources on the consumer's annual electricity costs generally increases with the 8 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 5. Conclusions The private savings that consumers can gain from their storage device will depend on the evolution of the electricity and energy systems. Consumers contemplating to invest in EES should not only be aware of the quantity of storage capacity deployed in the elec- tricity system but should also monitor the level of renewables that this aggregated storage capacity is likely to meet. This information is important because it affects the operational savings from storage by consumers, hence the probability of them investing in the technology. This could also be a reason for the complexity of cost- beneﬁt calculations by consumers and hence the current lack of EES deployments by domestic users [17,58]. This study investigates the potential economic savings to a UK electricity consumer as a function of energy storage coordination scheme, i.e., central vs. distributed, as well as the system-wide impact of deployment of such storage devices. As more con- sumers, and the wider electricity system, adopt electricity storage technologies, herding behaviour could occur: many cost- minimizing consumers with an incentive to shift electricity de- mand to the same periods of low electricity prices, which will ul- timately lead to an increased electricity demand and price peaks. Storage technologies already face multiple market barriers today. Hence, it is crucial to understand the impact of electricity market design on potential ﬁnancial beneﬁts of a storage owner (storer). Providing consumers with an understanding of how savings from their storage devices could be affected by numerous energy system conditions could improve consumer conﬁdence in the technology and might facilitate deployments. It is more likely for such information to be useful if provided in the form of a software integrated into an easily accessible website that calculates savings from storage based on high temporal and spatial resolution models of the electricity system. Such a model would consider where on the system the consumer is based, as well as the consumer's elec- tricity consumption patterns, among other factors. This would help inform the consumer's decision as to whether a ﬁnancial case to invest in storage exists in their speciﬁc case, and to understand the relationship between their investment on distributed technologies and their overall support for any future energy pathway. This paper examines the possible economic impact of owning a demand-side energy storage on the savings to a typical domestic consumer equipped with a solar PV microgeneration system. Table 3 When variable renewable capacity is large relative to ﬂexible supply capacity, there is a shortage of ﬂexibility on the supply-side, meaning that a system able to centrally coordinate more demand-side storage resources will be more valuable, and would produce more savings to consumers from their storage technology. Yet these insights must be checked against the possi- bility of distributed energy storage coordination to account for the likely scenario in which storage resources belonging to consumers are operated in a way that does not necessarily beneﬁt the system, so long as it beneﬁts the cost-minimizing consumer. The results also show the distributional effect of the centralized coordination on consumers. Those consumers owning ﬂexible technologies such as EES and providing the aggregator with the capacity of their device for load balancing, make relatively lower bill savings compared to those consumers with “No technology”. For example, PV-EES owners make 0e2% additional savings in the centralized scheme while consumers with no technology 2e10%. This is mainly due to the lower electricity prices for all consumers in the centralized coordination compared to a distributed sched- uling, which beneﬁts the most consumers under static tariffs with no technology. Therefore, the regulator should put a policy in place for redistributing some of the system-level beneﬁts back to the EES providers in the centralized coordination. In other words, the positive externality of aggregating distributed EES can be calcu- lated, including lower electricity prices at peak times and lower grid congestion management fees, and a part of that can be used to incentivize EES owners participating in the aggregation scheme. The lack of such incentives can deteriorate the economic attrac- tiveness of centralized coordination schemes for consumers [51e53]. Many consumers would prefer to dispatch their storage re- sources to reduce their own electricity bills rather than to reduce costs to the wider system. Hence, previous studies may have tended to overestimate the utility of storage in reducing electricity prices by assuming large amounts of demand-side energy storage aggre- gation. As additional storage capacity is deployed, the lower gap in peak and off-peak electricity prices diminishes the potential ben- eﬁts, sending a discouraging signal to the market for new in- vestments. Hence, policymakers should closely monitor the ﬂexibility requirements of the system and the willingness of 9 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Table 3 Energy 236 (2021) 121443 consumers could extract from their storage device by providing balancing or ancillary services to the grid have also been neglected. As electricity systems evolve, it will become increasingly important to assess the value of security and the potential provision of grid services through aggregation, as these are effectively substitutes to one another, while having synergies with energy arbitrage [23]. We simpliﬁed the representation of domestic consumers by consid- ering a typical domestic electricity consumer with a representative solar PV production and electricity consumption pattern. Yet these factors may largely vary across consumers and geographical areas. Furthermore, we focused on the role of coordination in the deter- mination of wholesale electricity prices. Yet to uncover the changes in retail tariffs, our modelling work would beneﬁt from an analysis where prices are made depending on capital, fuel, and networks costs in relation to each consumer in the electricity system. consumers to provide ﬂexibility services to the system. This can be done by internalizing the system-level beneﬁts of EES, through introducing incentives for investment in EES. From modelling method perspective, this implies that models of the electricity system should account for the trade-offs between private and system beneﬁts of energy storage aggregation. Yet it is unlikely that consumers will allow an aggregator to control their resources at all unless they are paid a ﬁnancial incentive to do so [57]. The decision by consumers to forego control of their storage resources could meaningfully reduce electricity wholesale prices [27]. This also entails the installation of smart meters and the access to the energy consumption data of private consumers, which they might be unwilling to share. The ability of aggregators and the System Operator to nudge consumers into providing such information could be key to the successful opera- tion of aggregators. 4.3. Additional storage in the electricity system and consumer savings y g y This paper further investigates the relationship between savings to a typical UK electricity consumer using energy storage only for arbitrage versus the amount of aggregated storage capacity deployed by the electricity System Operator. A ﬁve-fold increase in the level of aggregated storage capacity can potentially lead to 20% lower savings to the consumer from their energy storage device. We show that consumers should expect diminishing marginal savings to the private utility of their storage device because of additional aggregated storage capacity if they pay time-dependent electricity tariffs, such as dynamic ToU tariffs. To maximize the value of the storage resources, the system operator should reduce the uncertainty in investing in storage by providing the consumers with the information about amount of deployed storage resources in the system, either centrally or individually coordinated. The scale of reduction in electricity bills of consumers depends on future electricity system evolutions too. We demonstrate that a consumer could expect lower savings from their storage technology if a large amount of storage installed throughout the electricity system. Yet this only occurs if this ca- pacity is subject to aggregation. Annual electricity cost savings from storage to a typical UK consumer could fall by more than 20% if EES capacity were to increase from 3 GW to 17 GW in the system. The policy implication here is that the system operator should provide the data of the existing capacity of storage in the system, planned new storage installations, and the level of aggregation of these assets. This information should ideally be made public together with statistics about the fraction of these resources that are centrally coordinated as this is likely to impact the savings of consumers, lowering that compared to the case no storage de- ployments or aggregation occurred. 5. Conclusions We conclude that pairing solar PV with storage could reduce electricity bills for a typical UK consumer by 80e88%. 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This paper focused on arbitrage using EES, and the value of storage to consumers in providing non-economic beneﬁts such as energy security has not been considered. Similarly, the value that 10 B. Zakeri, G.C. Gissey, P.E. Dodds et al. Energy 236 (2021) 121443 [17] Hossain E, Faruque HMR, Sunny MSH, Mohammad N, Nawar N. A comprehensive review on energy storage systems: types, comparison, current scenario, applications, barriers, and potential solutions, policies, and future prospects. Energies 2020;13:3651. https://doi.org/10.3390/ en13143651. Author statement [22] Gardiner D, Schmidt O, Heptonstall P, Gross R, Staffell I. Quantifying the impact of policy on the investment case for residential electricity storage in the UK. J Energy Storage 2020;27:101140. https://doi.org/10.1016/ j.est.2019.101140. Conceptualization: B.Z., G.C.G., P.E.D., D.S. Methodology, Soft- ware, Data curation, and formal analysis: G.C.G., B.Z. Writing - Original Draft: B.Z., G.C.G. Writing - Review & Editing: B.Z. Visual- ization: B.Z., G.C.G. Supervision: P.E.D. Funding acquisition: G.C.G., P.E.D. [23] Pusceddu E, Zakeri B, Castagneto Gissey G. Synergies between energy arbi- trage and fast frequency response for battery energy storage systems. Appl Energy 2021;283:116274. https://doi.org/10.1016/j.apenergy.2020.116274. [24] Zakeri B, Syri S. Value of energy storage in the Nordic Power market - beneﬁts from price arbitrage and ancillary services. Int. Conf. Eur. Energy Mark. EEM 2016;2016e July. https://doi.org/10.1109/EEM.2016.7521275. Acknowledgments This research was funded by the UK Engineering and Physical Research Council (EPSRC) through the Realising Energy Storage Technologies in Low-carbon Energy Systems (RESTLESS) project (EP/N001893/1), for which the authors are very grateful. 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2011LIMO2001_113	French-Science-Pile	Open Science	Various open science	2,011	L'exotisme dans la littérature latine de Plaute aux écrivains augustéens	None	French	Spoken	1,363	2,138	tigre – tigresse : 33 (n. 3) ; 95-96 ; 104-105 ; 161 ; 185 ; 186 ; 187 ; 188 ; 189 ; 191-193 ; 195 (n. 1 et 2) ; 196200 ; 205 (n. 6) ; 208 ; 215 (n. 5) ; 235 ; 297 ; 344 ; 422 ; 512 ; 519 ; 553 (n. 3) ; 579 ; 585 ; 590-591 ; 593 ; 658 ; 662 ; 665 (n. 1) ; 669-670 ; 672 ; 695 ; 698 ; 699 (n. 4) ; 715 ; 716 ; 845 ; 889 ; 891 ; 892 ; 893 ; 904. Tigre (fleuve de Mésopotamie) : 138 ; 140-142 ; 158 (n. 1) ; 251 (n. 2) ; 512 ; 564 (n. 10) ; 847 ; 865 ; 877 ; 915. Timave : 138 ; 155 ; 580 (n. 3) ; 839-840. tissus et accessoires vestimentaires (voir aussi pourpre, soie) : 40 ; 42 ; 104 (n. 1) ; 223 ; 244 ; 246 ; 266-282 ; 347-352 ; 484 ; 490-491 ; 493 ; 494-495 ; 496 ; 506 ; 510-511 ; 520 ; 524 ; 526 ; 551-552 ; 555 ; 558 ; 559 ; 566 ; 568 ; 573 ; 594-595 ; 598 ; 604 ; 607 ; 611 ; 623 ; 639 ; 647-648 ; 650 ; 660-661 ; 691 ; 734 ; 735736 ; 737-738 ; 740-741 ; 742 ; 745 ; 747 ; 750-751 ; 753-754 ; 755-756 ; 757-758 ; 759 ; 763 ; 764 ; 768 ; 772-773 ; 774 ; 775-776 ; 815 ; 818-819 ; 851. tissus d’ameublement : 246 ; 287-291 ; 494 ; 520 ; 526 ; 551 ; 553 ; 588-589 ; 642 ; 650 ; 691 ; 716 ; 745 ; 747 ; 751 ; 759 ; 772 ; 818 ; 819-820. Tmolus : 160 ; 163 ; 229 ; 255 ; 518 ; 527 ; 540 ; 578. 25 Index thématique torque (collier celte) : 10 ; 345 ; 510-511 ; 554 (n. 2) ; 566 ; 612 ; 623 ; 654 (n. 4) ; 851 ; 873 ; 881 (n. 1) ; 886. 4) ; 73 (n. 2) ; 91 (n. 4) ; 99-100 ; 107 ; 168 ; 171 ; 172 (n. 5) ; 318 ; 340 (n. 3) ; 547 ; 557 (n. 2) ; 586 ; 612-613 ; 618 ; 642-643 ; 653-654 ; 665-668 ; 675 ; 708 ; 747 ; 756 ; 757 ; 758-761 ; 762 ; 764-765 ; 766 ; 903 ; 918. tragule : 354 ; 373. triomphe (cérémonie du) : 39 ; 43 ; 47 (n. 6) ; 104-105 ; 128 ; 142 ; 156 ; 184 ; 186 ; 187 ; 199-200 ; 212 (n. 1) ; 244 (n. 5) ; 282 ; 293 (n. 7) ; 294 (n. 6) ; 301302 ; 308 ; 310 ; 390 (n. 2) ; 506 ; 509 (n. 4) ; 511512 ; 519 (n. 4) ; 553 ; 555 ; 608 ; 616 ; 631 ; 634 (n. 6) ; 687 (n. 2) ; 689 ; 816 ; 818 (n. 2) ; 827 (n. 3) ; 830 ; 832-833 ; 835 (n. 3) ; 839 (n. 3) ; 841 ; 846 (n. 4) ; 847 ; 861 ; 863 (n. 6) ; 867-879 ; 880 (n. 1) ; 885 (n. 2) ; 889 ; 890 ; 891 ; 892 ; 894 ; 898 ; 900 (n. 3) ; 901. X Xerxès : 29 (n. 3) ; 132-133 ; 322 ; 332 (n. 1) ; 360 (n. 2 et 3) ; 410 ; 467-468 ; 485 (n. 4) ; 574 (n. 2) ; 625 ; 711 ; 713 ; 755 (n. 3) ; 814 ; 817 (n. 2) ; 831 ; 901 (n. 4). Triton (lac) : 114 ; 124 (n. 3) ; 326 ; 327 (n. 4) ; 543 (n. 3) ; 707 (n. 4). Troie – Troyens : 15 ; 24 ; 46 (n. 1) ; 78 ; 101 ; 108 ; 111 ; 127 ; 162 (n. 2) ; 163-164 ; 170 ; 171 ; 274 (n. 1) ; 281 (n. 5) ; 333 (n. 5) ; 347 (n. 9) ; 350-351 ; 394395 ; 400 ; 413 (n. 1) ; 462 ; 469 ; 482 ; 488-491 ; 494 ; 509 ; 552 ; 588 (n. 2) ; 597 ; 600 (n. 2, 4, 6 et 7) ; 601 ; 606-607 ; 618 (n. 5) ; 750 ; 763 ; 775-776 ; 810 ; 848 ; 908-909 ; 915-916 ; 917 (n. 4) ; 918. trophées et monumenta : 39 ; 46-47 ; 128 ; 171 (n. 6) ; 293 ; 354 ; 509 ; 801 ; 829 ; 831 (n. 7) ; 832-833 ; 834 ; 843 (n. 3) ; 867 ; 868 ; 879-888. Tyr et Sidon : 79 (n. 4) ; 113 170-171 ; 174 ; 175 (n. 4) ; 269 (n. 6) ; 270 (n. 5) ; 271-276 ; 315 (n. 2) ; 318 (n. b, cf. note complémentaire, p . 921) ; 319 ; 321 (n. 2) ; 350 ; 406 ; 459 ; 469 ; 471-472 ; 476 ; 494-495 ; 533 ; 538 ; 542 ; 574 ; 597 ; 598 ; 609 ; 626 (n. 5) ; 660 ; 663 (n. 1) ; 681 (n. 1) ; 715 ; 742 ; 751 ; 755756 ; 757 ; 758 ; 759 ; 761 (n. 1) ; 764 ; 768 ; 820 ; 841 ; 860 (n. 4) ; 898 ; 908 (n. 3). U Ulysse : 1 ; 22 ; 24 ; 71 ; 74 (n. 2) ; 85 ; 93 (n. 3) ; 596 ; 599 ; 607 ; 692 (n. 4). unicorne (ou licorne ; animal fabuleux) : 219 (n. 2). urus (auroch) : 221-222 ; 382. V Vercingétorix : 638 (n. 1) ; 849 ; 853 (n. 1 et 3) ; 856. viandes exotiques : 214 ; 237-238 ; 242-244 ; 260 ; 745 ; 753 ; 769. voyage : 1-3 ; 4 ; 5 ; 12 ; 18 ; 22 ; 24 ; 27-28 ; 29 (n. 4 et 5) ; 33-36 ; 38-40 ; 52 (n. 2) ; 69 (n. 1) ; 70 ; 72 (n. 26 ERRATA Suite à un problème de traitement de texte qui n’a pu être résolu au moment de l’impression de cette thèse, certaines notes de bas de page se trouvent reportées sur la page suivante. Par ailleurs, nous avons pu, à ce jour, effectuer les corrections suivantes : p. 17, n. 2, l. 14 : engagée par les intrigues p. 22, l. 26-27 : des Premiers livres Académiques. p. 99, l. 13 : est d’ailleurs attribuée p. 126, n. 2, l. 4 : de l’Hister p. 170, n. 2, l. 7 : devant ses troupes qui craignent p. 247, l. 3 : nec miserat India costum p. 261, n. 2, l. 1 : déjà employée chez Plaute p. 264, l. 14 : semble avoir été connue p. 292, n. 9, l. 2 : en Éthiopie p. 304, n. 2, l. 5 : pour les entretiens philosophiques p. 353, l. 12 et p. 389, l. 4 : dans ses Didascalica. p. 375, n. 6, l. 1 : mais tu es p. 449, l. 5 : en comparant Lycé p. 453, n. 2, l. 11 : l’allusion à l’immanitas p. 457, l. 4 : épithète traditionnellement appliquée p. 487, n. 3, l. 3 : dans les Tusc., V, 35, 101, où Cicéron ajoute... p. 498, n. 4, l. 4 : gaieté p. 602, l. 19 : contenir des allusions. p. 654, l. 4 : de nombreux passages relatifs p. 682, n. 1, l. 10 : par approximation analogique ; n. 6, l. 1 : le premier terme p. 757, n. 5, l. 3 : où Properce... ; l. 5 : Marius et Jugurtha p. 816, n. 1, l. 6 : on était allé Nous vous prions de bien vouloir nous excuser pour les désagréments occasionnés par ces erreurs ou omissions, ainsi que par celles qui n’auraient pas été répertoriées.	557
https://openalex.org/W1577211016_5	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	7,105	13,039	“… no sólo los colonizadores sino también los colonizados deben aceptar el estilo de vida de <los de arriba> como el único modelo de la buena vida…. Este proceso va acompañado invariablemente de una devaluación de los propios: de la propia cultura, del propio trabajo, tecnología, estilo de vida, y a menudo también de la propia filosofía de la vida y las propias instituciones sociales…. Los colonizadores imponen esta devaluación al principio por la fuerza y luego la refuerzan a través de la propaganda, de programas educativos, de una modificación de leyes y de la dependencia económica, como resultado de la trampa del endeudamiento, por ejemplo. Finalmente, los colonizados a menudo acaban aceptando e interiorizando esta devaluación como si fuese la situación <natural>. Uno de lo problemas más difíciles para los colonizados (países, mujeres, campesinos) después de un proceso formal de descolonización es el desarrollar su propia identidad: una que no este basada en el modelo del colonizador como la imagen del verdadero ser humano”. MIES, MARIA. “El mito de la recuperación del retraso en el desarrollo”, en MIES, MARÍA. SHIVA, VANDANA. Ecofeminismo. Teoría Crítica y Perspectivas, op. cit., p. 89. 154 PLUMWOOD, VAL. Feminism and the Mastery of Nature, op. cit., pp. 59-60. 155 FREIRE, PAULO. Pedagogia do Oprimido, Ed. Paz e Terra, 1970, p. 22. Traducción al castellano de Jorge Mellado, Pedagogía del Oprimido, Siglo XXI, 2005. 153 70 esfera pública del poder y de la razón en la medida de su asimilación de los valores masculinos (frialdad, impersonalidad, distanciamiento, ausencia de compasión, etc.). Sin embargo, esta reflexión de las características estructurales del dualismo es necesaria para superar la identidad dualizada, siendo uno de los aspectos más importantes para la reconstrucción de las relaciones y de la identidad en términos de un concepto no jerárquico de la diferencia. Las sugerencias de Val Plumwood para que se desarrolle una apropiada relación de la diferencia no jerárquica tendría las siguientes características específicas: Segundo plano (negación) - un concepto no-jerárquico de la diferencia requiere un cambio en los pensamientos, consideraciones, percepciones y tomas de decisiones que reconozcan la contribución de lo que ha sido considerado segundo plano, y que reconozca la dependencia creada por estas relaciones. Exclusión Radical - un concepto no jerárquico de la diferencia afirma la continuidad, replantea relaciones de formas más integradas y rompe la falsa elección híper separada presente, mediante la recuperación de la esfera que fue negada. Incorporación (definición relacional) - un concepto no jerárquico de la diferencia debe revisar la identidad de los dos lados: inferior y superior. Esto puede tener el propósito de redescubrir el lenguaje y la historia del lado inferior, reclamar fuentes de identidad positiva e independiente y afirmar la resistencia. Instrumentalismo - un concepto no jerárquico de la diferencia implica reconocer al otro como un centro de necesidades, valores y esfuerzos por su propia cuenta, un ser cuyos fines y necesidades son respetados e independientes. Homogeneización - un concepto no jerárquico de la diferencia supone reconocer la complejidad y la diversidad de otros, por ejemplo de “otras naciones” que fueron homogeneizadas y marginalizadas en su constitución como “los otros excluidos”, como “el resto”. Estas respuestas corresponden a una síntesis de las principales preocupaciones culturales y conceptuales de varios movimientos de liberación. De esta forma, para establecer claramente lo que está implicado en el dualismo, tenemos que mirar las señales que apuntan la dirección de la salida y es esta la intención de deconstruir la historia del dominio. 71 2.4. Implicaciones de los dualismos Para Plumwood no sólo la identidad masculina en si es subrayada en la concepción platónica de la razón y de la vida de la razón, también la identidad del amo, del señor, del dominador, definida en términos de múltiples exclusiones, y en términos de dominación no solo del femenino sino también del esclavo (lo que usualmente combina opresiones de raza, clase y género), del animal y de lo natural. Conjuntamente con otras teóricas, Plumwood también amplía la perspectiva crítica feminista del tratamiento platónico previamente aplicado a las mujeres y a la razón, apuntando que en la visión del mundo de Platón encontramos una importante fuente del dualismo razón/naturaleza y que el tratamiento que él aplicaba a la naturaleza tiene un importante paralelo con el de las mujeres156. Varios historiadores medioambientales contemporáneos157 han revisado la cultura clásica y han encontrado en la filosofía medioambiental de los griegos un modelo potencial y correctivo para nuestra propia sociedad y tiempo, retratando la sociedad griega como amistosa con el medioambiente, que valoraba las pequeñas escalas, que veía la vida humana como perfectamente sujeta a control y a límites, y que enfatizaba las consecuencias adversas de la arrogancia humana. Se retrata a los griegos respetando y celebrando la naturaleza a través de la adoración de Gaya, la antigua diosa de la tierra. El historiador J. Donald Hughes, que trata la tradición griega en estos aspectos158, amplia a Platón la favorable valoración de las actitudes griegas hacía la naturaleza. Cita la discusión de Platón sobre la deforestación de Ática en Timeo159, y presenta las referencias de Platón sobre Gaya y sobre el alma del mundo como indicativo de una forma orgánica de verlo, como una forma de decir que necesitamos de la 156 IRIGARAY, LUCE. Speculum of the other woman, trans. Gillian C. Gill, Ithaca, NY: Cornell University Press, 1985; HARTSOCK, NANCY C. M. Money, Sex and Power, Northeastern, 1985, op. cit.; RUETHER, ROSEMARY RADFORD. New Woman, New Earth: Sexist Ideologies and Human Liberation, Seabury Press, 1975: New York; SPELMAN, ELIZABETH. The Inessential Woman, Boston, Beacon, 1988. 157 NAESS, ARNE. “Man apart and deep ecology: a reply to Reed”, Environmental Ethics, 1980 pp. 183-92; HUGHES J. DONALD. “Gaia: an ancient view of our planet”, Environmental Review 6, 1982; SALE, KIRKPATRICK. Dewellers in the Land: The Bioregional Vision, Sierra Club Books, 1985, San Francisco; TANNER, R.G. “Environmental Insight in Greek philisophy in antiquy”, Proceedings of Ecopolitics V, Sydney: Centre for Liberal and General Studies, University Of South Wales, 1992 AU; PLUMWOOD, VAL. “Prospecting for an Ecological Gold Among the Platonic Forms”, 1997, Ethics and Environment, 2 (2), pp. 149168; PLUMWOOD, VAL. Feminism and the Mastery of Nature, op. cit., p.73. 158 HUGHES J. DONALD. “Gaia: an ancient view of our planet”. Environmental Review, 1982, p. 6. 159 HUGHES J. DONALD. “Gaia: an ancient view of our planet”. op. cit., p. 7. 72 naturaleza, una forma no muy distinta de la que encontramos hoy en la Hipótesis de Gaya160 que ve el mundo como un organismo vivo. Plumwood argumenta que este retrato de Platón como un ambientalista es erróneo, pues, por ejemplo, en La República161, en la visión de la tierra como madre -en la que se basa este favorable retrato- está claro que él considera falsa esta historia, pues la coloca en la categoría de mentiras útiles (o “mitos magníficos”) que los guardianes dominantes pueden propagar si les sirve para fines sociales. Val señala que la discusión de Platón sobre el mito de la Madre Tierra precede inmediatamente a su discusión sobre las clases de bronce, oro y plata, una historia que él sugiere como útil para estimular la estratificación social, y que, similarmente, la historia de Gaya es presentada como falsa pero útil porque anima a los ciudadanos a luchar por el Estado como si fuera por su propia tierra162. Para Plumwood lo que el mito conduce a promover no es el ambientalismo, sino el militarismo. En sus palabras, Platón es un candidato poco probable a la canonización tanto como protector del medioambiente, como de las mujeres163. Para esta autora, considerar que la filosofía da Platón es favorable al medio ambiente y, además feminista, es un gran error y justifica tal aserción analizando diversos pasajes de sus escritos en los que considera la tierra y la naturaleza como instrumentalizadas y como un mero medio para los fines humanos, que son los únicos seres dignos de consideración moral. Para Val Plumwood Platón es el mentor filosófico y conceptual de la perspectiva de la tierra y del mundo natural como “un vertedero”, como naturaleza no humana y sin forma, caótica y vacía, sin fines en sí misma, desprovista de cualidades, y como receptáculo pasivo de la forma superior dada por la razón en un proceso en que no hay ningún elemento de mutualidad. Dentro de este análisis con una perspectiva de género, la consistente negación y derogación de las mujeres y de las características consideradas femeninas no es meramente una “misoginia casual”, sino un elemento fundamental y un marco extremadamente influyente de la filosofía de Platón, que asocia persistentemente las mujeres y sus vidas con el lado inferior del ser, y el tratamiento de la razón y de la filosofía como opuesta y excluyente del orden inferior. Dentro de este análisis, el “feminismo” atribuido a 160 LOVELOCK, J. E. Gaia, una nueva visión de la vida sobre la tierra. Hermann Blume, 1983. LOVELOCK, J. LYNN, MARGULIS. ATLAN, F. VARELA. MATURANA, H. Y OTROS. Gaia. Implicaciones de la nueva biología. Editorial Kairós, 1989; LOVELOCK, J. Las edades de Gaia. Editorial Tusquets, 1993; LOVELOCK, J. La venganza de la tierra: por qué la tierra está rebelándose y cómo podemos todavía salvar la humanidad. Editorial Planeta, 2007; LOVELOCK, J. Homenaje a Gaia. La vida de un científico independiente. Editorial Laetoli, 2005. 161 LEE, DESMOND. Plato: Timaeus and Critias, Penguin/Mx, 1987, p.181. 162 Ibidem. 163 PLUMWOOD, VAL. Feminism and the Mastery of Nature, op. cit., p.73. 73 Platón164 supone la fuerte devaluación de la mayoría de las mujeres y de sus vidas, pues solo la elite femenina que había sido educada con éxito en los cánones de la razón estaban entre las guardianes de la Kallipolis. Las mujeres participaban en función de sus semejanzas con los hombres, no de sus diferencias. Val Plumwood considera que la esencia de la teoría de Platón es la separación radical entre el orden superior del logos y el orden inferior del caos y entre las esferas racional y material, explicándolas a través de un conjunto de metáforas que enmarcan múltiples exclusiones: maestro y esclavo, hombre y mujer, alma y cuerpo, y el dominio y control de la parte inferior por la superior. Bajo esta mirada, se puede concluir, conjuntamente con Plumwood, que Platón anticipó los principales aspectos del cristianismo y presentó la primera y duradera declaración de los principios del paraíso de la historia occidental. El filósofo proporcionó una consideración inmensamente influyente de la identidad humana, del significado de la muerte y de la relación del alma y el cuerpo. Las implicaciones ecológicas de la doctrina de Platón, que considera al hombre como celestial y no como terrestre, son profundas, pues establece de forma explícita y teórica una consideración del significado y real localización de la vida humana como más allá de la tierra. La tierra no es un hogar para ser apreciado, sino un ensayo, una prueba, un lugar de paso temporal y de significado menor en comparación con el mundo del más allá. Val Plumwood considera que Platón anticipa la posterior negación de Descartes de la dependencia de los sentidos y su forma de tratarlos como fuente de error, así como su desarrollo del dualismo humano/naturaleza en términos del dualismo mente/cuerpo165. Plumwood nos enseña, por tanto, que el primer racionalismo de Platón construyó el genuino ser humano en términos polarizados y opuestos, desprovistos de las cualidades de los animales y de la esfera de la naturaleza, y el siguiente, el de Descartes166, construyó la naturaleza privada de las cualidades propias del hombre. De esta forma el desarrollo del dualismo humano/naturaleza destruye las características compartidas por ambos. Fueron los antiguos racionalistas Cartesianos los que declaran la naturaleza no humana como terra nullis, desprovista de la mente, como un terreno baldío totalmente disponible para la anexión, una esfera fácilmente moldeada por los fines de una razón concebida como ilimitada. 164 ANNAS, JULIA. “Plato’s Republic and Feminism”, Philosophy 51, 1976; NANCY VANHEERST. “Feminism and Plato”. Ancient Philosophy, 1999, entre otras. 165 PLUMWOOD, VAL. Feminism and the Mastery of Nature. op. cit., pp. 69-103 166 Ibidem, pp. 104-119. 74 Con estos elementos se puede concluir que en la evolución del dualismo humano/naturaleza, el primer paso fue la elaboración de la identidad humana como mente o razón, excluyendo o interiorizando la rica gama de otras características humanas y no humanas o construyéndolas como no esenciales. El segundo paso fue la creación de la mente o razón en términos exclusivos y en oposición a la naturaleza. La construcción de la naturaleza en sí misma como inconsciente es el tercer paso. Los primeros dos pasos señalados anteriormente están explícitos en Platón, y el tercero está explícito en Descartes en su tratamiento de la materia original como caos, la materia inconsciente o la forma primitiva del mundo en el que el orden racional debe ser impuesto. El último paso, que Descartes hace explícito, se considera a menudo como el elemento problemático introducido por la Ilustración y para Plumwood hubo una mayor intensificación del dualismo humano/naturaleza en esta época. Siguiendo en este análisis de la filosofía europea occidental, Plumwood pasa a demostrar como Locke167 enseña como la razón, en un individuo autónomo y racional, puede rentablemente apropiarse de toda la naturaleza como si fuera su propia propiedad, tras desposeer y hacer invisibles a los primeros habitantes y su prioridad de los derechos, cuya agencia es excluida por los conceptos eurocéntricos de productividad y trabajo en los procesos de colonización168. Para la autora australiana esta etapa del desarrollo del dominio acentúa la concepción de la racionalidad como egoísmo y de la naturaleza en términos instrumentales y como un recurso para el dominador. Esta es la etapa de la búsqueda y explotación de la utilidad en (o del uso de) la naturaleza, en la que el ego racional es concebido como separado del “otro” alienado, como el auto suficiente usuario de artículos de consumo o de recursos para el conocimiento, en lo que ahora se ha convertido la naturaleza. Estas exclusiones contrastantes incluyen también la ética y la belleza, además de las ideas de la esfera privada, de la casa y de lo femenino, del altruismo, de los valores, de las emociones, de las relaciones y del cuidado. La esfera privada recibe legitimación en la medida en que puede ser contenida e instrumentalizada, hecha para servir a la esfera primaria del orden racional; el dominio de lo 167 Lo mismo afirma Vandana Shiva, respeto a Locke: “Así, en cuanto saca del estado en que lo ha producido y dejado la Naturaleza, (el hombre) incorpora su trabajo y de este modo se lo apropia” como si fuera suyo. LOCKE, JOHN. Two Treatises of Government. J. M. Dent & Sons, Londres, 1991, mencionado en SHIVA, VANDANA. “Reduccionismo y regeneración: crisis en la ciencia”, en MIES, MARÍA. y SHIVA, VANDANA. Ecofeminismo. Teoría Crítica y Perspectivas, op. cit., pp. 44, 54. 168 PLUMWOOD, VAL. Feminism and the Mastery of Nature. op. cit., pp. 111, 117-9. 75 privado y de lo femenino no tiene condición política en sus derechos y puede ser pensado solo si es reducido y sometido bajo el liderazgo de la partícula primaria. Dando un salto en el tiempo en la Tesis del Dominio, la autora pasa a discutir las modernas teorías económicas y liberales. Val apunta que en estas teorías, la nueva concepción de la razón continúa negando y poniendo en segundo plano todas las áreas previamente excluidas y consideradas como naturaleza, tratándolas en forma de aportaciones invisibles a la economía racional. Se añade a esto un intenso instrumentalismo El sujeto económico cada vez más activo, y una nueva forma de dominio trata de definir la racionalidad como egoísmo y la sociabilidad como una asociación instrumental dirigida por intereses propios. En consecuencia, el dualismo yo (self)/otros se convierte en la clave para los conceptos de individualidad y racionalidad. Analizando las aportaciones de Kant, Plumwood describe que ahora el dominio de la ética es el dominio de los que no han sido instrumentalizados, cuyas necesidades y agencia son las únicas tomadas en consideración. Kant utiliza la razón para justificar la división entre los medios y los fines, o entre los instrumentalizados y los que instrumentalizan. Los seres cuya existencia no depende de nuestra voluntad, sino de la naturaleza, tienen solo un valor relativo cuando se trata de seres irracionales, y por esto se llaman cosas; pero los seres racionales se denominan personas, porque su naturaleza ya los señala como fines en sí mismos, es decir, como algo que no puede ser usado como medio y por lo tanto se impone un límite a todo uso arbitrario de tales seres, que son por tanto objetos de respeto.169 En el actual mercado económico liberal las líneas entre el humano utilizador y el no humano utilizado no están, en la práctica, tan claramente marcadas como en la historia oficial liberal del siglo XVIII analizada, y el tratamiento instrumental en la esfera humana es todavía más invasivo. En la vida moderna la naturaleza, las mujeres y todos los demás grupos subordinados siguen siendo instrumentalizados, pues son inscritos en este particular tipo de identidad. Sin embargo el egoísmo y sus cualidades asociadas, como la asertividad y la competitividad (que forman parte de las cualidades tradicionalmente consideradas propias del hombre) son ahora asumidas por las instituciones de la esfera pública y por las grandes corporaciones. 169 51. KANT, IMMANUEL. Fundamentación de la metafísica de las costumbres. Santillana, Madrid, 1996, pp. 50- 76 Como hemos visto, la sedimentación cultural de estos valores de dominación y superioridad se ha desarrollado a través de diversas narrativas adaptadas a cada momento histórico, de tal forma que estos valores fueron incorporados por toda la humanidad (o casi toda) como absolutos. La cultura del dominio, en mi entender, una cultura de falta de respeto hacía los demás y falta de reconocimiento de la fragilidad y dependencia de la vida humana, es la gran responsable de la actual crisis ambiental y social. En otras palabras, el actual caos en que vivimos es una Crisis Ecológica de la Razón, como veremos seguidamente. 77 CAPÍTULO III RACIONALISMO Y LA AMBIGÜEDAD DE LA CIENCIA 78 3.1. La Crisis Ecológica de la Razón The deterioration of the global ecological context of human life demands from our species a clear and adequate response, but we are seemingly immobilized, even though it is clear that at the technological level we already have the means to accomplish the changes needed to live sustainably on and with the earth. So the problem is not primarily about more knowledge or technology; it is about developing an environmental culture that values and fully acknowledges the nonhuman sphere and our dependence on it, and is able to make good decisions about how we live and impact on the non-human world.170 Como hemos podido comprobar en el capitulo II, gran parte del trabajo e investigación de la mayoría de las feministas ecológicas críticas intentan deconstruir las formas de pensamiento racionalizado e instituido desde su fundación, tratando de buscar referencias en la historia y en la filosofía clásica, y desafiando radicalmente el paradigma cultural para (re)leer y (re)contar tanto la historia como los principios teóricos que cimentaron nuestra vida social y cultural. En esta re-lectura estas teóricas intentan encontrar las brechas, las entrelíneas y las tendencias que fueron consideradas neutrales pero que modelaron la razón como forma de distinción y de exclusión. Me atrevería a decir que, desde mi punto de vista, esta es la verdadera e importante revolución intelectual, cultural y académica de nuestra época171 (conjuntamente con la revolución digital) y que es consecuencia directa del fortalecimiento de los grandes movimientos sociales que se desarrollaron más fuertemente en la segunda mitad del siglo pasado. Ahora, esta corriente revolucionaria viene a decirnos –aun que implícitamente– que es hora de recontar la historia, de revisar nuestras concepciones sobre todo lo que fue y es considerado como válido, bueno y justo y que se ha dado por sentado durante toda la evolución del pensamiento occidental –tal y como lo concebimos-. Es hora de re-ubicar todos nuestros héroes –principalmente los héroes de guerra, que se hicieron ricos y crearon imperios tras la invasión, anexión, violación, usurpación, engaño, pillaje, uso de la 170 PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 3. Explico: digo esto, aún siendo consciente de que desde siempre hubo muchas voces contrarias a los movimientos y corrientes políticas dominantes. Lo que pienso es que, ahora, como jamás antes, tenemos derecho de hablar y de pensar y con todo un sistema que no sólo facilita, sino que permite alcanzar grandes distancias y distintas poblaciones, lo que hace con que asuma dimensiones inimaginables. 171 79 fuerza extrema y de exterminio en contra poblaciones desprotegidas y en condiciones sabidamente más vulnerables, y también es hora de rediscutir y reconsiderar nuestros valores, pues es inequívoco que hemos llegado a un punto alarmante tanto ecológica como socialmente172. En esta parte de la Tesis pretendo comprender las principales críticas y denuncias elaboradas por las investigaciones ecofeministas y feministas críticas, que apuntan de forma radicalmente critica, objetiva y categórica a los daños causados por el uso sin criterio de la ciencia y de las nuevas técnicas del mercado económico global y que nos presentan un rico panorama del tema. Tanto Val Plumwood como Vandana Shiva, presentan una dura y radical crítica del mundo considerado desarrollado y muestran lo que sucede en sus países –Australia e India, respectivamente- y en los países considerados como libres para la apropiación, por el hecho de ser “pobres”, subdesarrollados” o menos racionales173. Las dos autoras denuncian duramente las técnicas invasivas y sobre-explotadoras de pesca, la contaminación del suelo, del aire, de las aguas y de todo el planeta, tras el empleo de las técnicas de la agricultura 172 En las clases de Políticas Ambientales ministradas por el Profesor Viriato Soromenho-Marques, del curso del invierno de 2013, de la Facultad de Filosofía de la Universidad de Lisboa, hemos analizado una serie de artículos muy interesantes y actuales que, aunque no sean académicos, demuestran de forma inequívoca que la preocupación hacia el medio ambiente es generalizada y afecta las más diferenciadas ramas de la cultura. Estos artículos fueron publicados por la revista americana TIME, desde los años 70 y son apenas algunos pocos ejemplos de lo mucho que si se viene hablando sobre los problemas y impactos ambientales desde los años 60, momento en que siquiera teníamos el medio ambiente como una rama específica del Derecho. Cada uno de ellos –de estos artículos– dan la sensación de que fueron recién escritos, en función de la actualidad de los temas y de las formas con que son abordados, lo que puede darnos, mas que nada, la sensación de que mucho se ha dicho, y sigue siendo, pero muy poco, de hecho, si ha logrado, debido a la evidente constatación de que desde entonces – de los años 60– los daños ecológicos se han multiplicado de forma alarmante y global. Revista TIME: “Fighting to sabe the Earth”, Monday, Feb 2, 1970; “Environment: Shrinking Shores”, Monday, Aug. 10, 1987; “Environment: The Dirty Seas, Monday, Aug. 01, 1988; “Planet Of The Year: What on EARTH Are We Doing?, Monday, Jan. 02, 1989; LINDEN, EUGENE. “Environment: Owl vs Man”. Monday, Jun. 25, 1990; “Lost Tribes, Lost Knowledge, Monday, Sep. 173 Gran parte de la lucha de las Ecofeministas teóricas y activistas es cambiar el concepto de “pobreza”, en el sentido de que no se puede considerar pobre una cultura que tenga tierras fértiles y agua potable por ejemplo y, dentro de este contexto, denunciar las grandes empresas de la economía globalizada que están convirtiendo estos países en sus basureros (como el caso de países de África que se han convertido en basureros nucleares), o en los que las multinacionales se instalan para apropiarse de sus recursos, como agua, minerales etc. MIES, MARÍA. y SHIVA, VANDANA. Ecofeminismo. Teoría Crítica y Perspectivas, op. cit., pp. 88, 112-35; BOSERUP, ESTER. Women´s Role in Economic Development. Allen and Unwin, Londres, 1960; DAWN. Development, Crises and Alternative Visions: Third World Women´s Perpective. Christian Michelsen Institute, Bergen, 1985. 80 moderna, de la gran concentración en las grandes ciudades, además de la insistente destrucción de los bosques tropicales. Después de presentar cómo se ha desarrollado la historia de la dominación en el mundo en los capítulos anteriores, ahora se pretende indagar cómo estos valores se asentaron y se desarrollaron en las nuevas economías de mercado, institucionalizando el poder en sus vertientes más agresivas. Siguiendo con la elocuente crítica de Val Plumwood, veremos cómo se implantaron estos valores e instituciones como herederos del sistema del dominio y que nos han conducido a lo que ella denomina la crisis ecológica de la razón. Tales valores son el racionalismo y el centrismo, la ciencia y su ambigüedad, el desarrollo del antropomorfismo y del antropocentrismo, y la mercantilización. Como nos ha dicho Gómes-Hervas, con el super hombre nietzscheano como contrafondo, el antropocentrismo moderno ha podido ser interpretado como el símil del “complejo de llegar a Dios, a través de la ciencia que la convierte en omnisciente y a través de la técnica que la convierte en omnipotente174. Como el núcleo del análisis de Val Plumwood es la crisis ecológica en el mundo y la cultura occidental, obviamente, uno de sus puntos de partida es la ciencia, donde afirma que la ciencia se ha convertido en “una forma de pensamiento monológico y dualístico”, donde los científicos se posicionan como “completamente apartados de los objetos en estudio, de tal forma que les niegan la capacidad mental o la intencionalidad”175. De diversas formas bastante convincentes, ambas teóricas mencionadas afirman que, bajo esta mirada extremadamente racionalista, la naturaleza se ha convertido en un objeto totalmente cognoscible y tecnológicamente manejable, además de perfectamente reproducible y reemplazable. Tal pensamiento es altamente anti-ecológico, pues ignora la interconexión entre los humanos, los no-humanos y la naturaleza, además de aumentar la preocupación respeto de la mercantilización de la ciencia, pues la adquisición del conocimiento esta cada vez más justificada por motivaciones instrumentales176. Val Plumwood traza una parodia sobre los auto-denominados Eco-Guardianes, en una alusión a los Guardianes de La República de Platón, tratándolos como si fueran los supuestos científicos ambientales del primero mundo, comprometidos en salvaguardar el 174 GOMEZ-HERAS, JOSÉ M.ª GARCÍA. En armonía con la naturaleza: reconstrucción ambiental de la filosofía. Madrid, Biblioteca Nova, 2010, p. 443 175 PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 45. 176 SHIVA, VANDANA. “Reduccionismo y regeneración: crisis en la ciencia”, en MIES, MARÍA. y SHIVA, VANDANA. Ecofeminismo. Teoría Crítica y Perspectivas, op. cit., pp. 39-42, 71-2, 102. 81 planeta y la naturaleza. Pero ellos no tienen la capacidad de reconocer su propio conocimiento como políticamente situado y, por lo tanto, fallan en reconocer la necesidad de ser socialmente inclusivos y activamente comprometidos en reconocer limites y exclusiones. Sus posiciones privilegiadas les permiten olvidar completamente el hecho de que sus conocimientos son producidos dentro de una estructura social desigual. A través de esta parodia del racionalismo, Plumwood intenta promover una apropiada “racionalidad ecológica”, que conduciría a una ciencia más auto-crítica, plenamente consciente y sensible a su papel activo y conjunto con la sociedad177. Pero su crítica va más allá de la defensa de la justicia social e incluye, también en la misma medida, a la naturaleza no-humana, para desarrollar una exhaustiva y sofisticada crítica del antropocentrismo. Ella afirma que la visión occidental del mundo basada en la razón, esta dividida en dos reinos: de un lado el activo, conocido como “sujeto”, y del otro el pasivo, reconocido como “objeto”. Como resultado, se produce una discontinuidad radical entre los humanos (como los únicos posesores de la razón) y los no-humanos178. A continuación, la autora apunta algunas formas en que el dualismo centrado en los humanos es inherentemente anti-ecológico y arrogantemente radical, como lo es el fracaso de la ética en considerar las limitaciones de la especie humana y la homogenización de toda la naturaleza como inconsciente y, por lo tanto, indigna de consideración moral. Además, esta negación o menosprecio de la dependencia humana de la naturaleza, ha convertido a ésta en un mero problema tecnológico a ser superado179, y como consecuencia, se desarrolla un falso sentido de la autonomía humana. El resultado es que la naturaleza se convierte en un espacio puramente negativo –sin cultura (como ya se ha comentado anteriormente, la naturaleza no es considerada como parte de la cultura en el dualismo cultura/naturaleza), sin desarrollo, sin nada en si misma y, finalmente, su valor solo puede ser considerado en función de los valores e intereses humanos180. Plumwood afirma, enfáticamente, que estos fallos deben de ser 177 PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., pp. 68-70. Ibidem, p. 100. 179 Ibidem, p. 105. 180 BENHABIB, SEYLA. “The generalised and the concret other”, in Seila Benhabib and Drusilia Cornell (eds) Feminism as critique, Polity Press 1987. Cambridge; BENJAMIN, JESSICA. “The bonds of love: rational violence and erotic domination”, in Hester Einsenstein and Alice Jardine (eds) The Future of Difference, New Brunswick, Rutgers University Press 1985, New Brunswick; BENJAMIN, JESSICA. The bonds of love: feminism and the problem of domination, Virago, 1988. New Brunswick, NJ; CHENEY, JIM. “Ecofeminism and deep ecology”, Environmental Ethics vol. 9, 1987; CHODOROW, NANCY JULIA. “Gender, relation and difference in psychoanalytic perspective”, in Hester Einsenstein and Alice Jardine (eds) The Future of Difference, New Brunswick, Rutgers University Press 1985, New Brunswick, NJ; GRIMSHAL, JEAN. Feminist Philosophers, Wheatsheaf, 1986. Brigton, Sx; HARTSOCK, NANCY C. M. Money, Sex and Power, Northeastern, 1985, p.77, Boston, MA; POOLE, ROSS. Morality and Modernity, Routledge, 1985, London; 178 82 superados si queremos tener cualquier esperanza tanto para nuestra supervivencia como para la de la naturaleza181. En las obras aquí analizadas, estas brillantes feministas teóricas y activistas invitan a una radical transformación de nuestra forma de ver el mundo, con ojos profundamente comprometidos con la defensa de la justicia social, con la naturaleza y con el feminismo. Partiendo del punto exacto en que se termina el análisis de la historia del dominio en el planeta, lo retomamos ahora para ver cómo el razonamiento sigue en esta misma trayectoria -la del dominio– solo que ahora con los refinamientos de una razón más sofisticada, globalizada y científica. Los procesos contemporáneos de destrucción ecológica en los bosques, el suelo, el aire y los océanos parecen darnos una clara advertencia de estos puntos ignorados en nuestra economía racionalista. Las principales corrientes y teorías del feminismo ecológico crítico hicieron y siguen haciendo un profundo y critico análisis de los principales daños causados en la naturaleza tras los modernos métodos invasivos de la tecnología racional. Sus principales críticas se dirigen a la absoluta falta de visión – política, social y jurídica – de los impactos altamente nocivos con que las modernas tecnologías están destruyendo la naturaleza en nombre del desarrollo y del aumento de la productividad. Los estudios comprueban, por ejemplo, los efectos en gran escala del mercado global y de la economía pesquera en las poblaciones mundiales de peces: todos los océanos del Planeta están contaminados y el 75% de las poblaciones de peces del mundo son reconocidamente sobreexplotadas o se acercan de la sobreexplotación, restando solamente un 25% que no están amenazadas182. La economía pesquera global es insostenible pues esta fallando al no prevenir y/o impedir un colapso ecológico183. WARREN, KAREN. “The power and the promise of ecological feminism”, Environmental Ethics 12 (2), 1990; PLUMWOOD, VAL. Feminism and the Mastery of Nature, op. cit., p. 151. 181 Ibidem, p. 122. 182 FAO, FISHRIES REPORT N. 638, FAO, 2000, New York, Papers presented at the Expert Consultation on Economic Incentives and Responsible Fisheries, 28th November – 1st December 2000. FAO FISHRIES REPORT SUPPLEMENT N. 638, 9600., ISBN 92-5-104554-2 – FAO Publications, en PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 25. 183 En España y en toda la costa Atlántica la situación no es muy diferente. Un informe publicado en agosto de 2012 por NEF (New Economics Foundation) y OCEAN2012 revela que si la población española solamente consumiera pescado de suministro propio, se quedaría sin pescado el 8 de mayo, pasando a depender del pescado procedente del resto del mundo a partir del 9 de mayo, sobre la base de los niveles de consumo actuales. Los medios de comunicación y los Activistas Ecológicos de diversas vertientes no se cansan de advertir y manifestarse en contra de la sobrexplotación oceánica. Esta sobrexplotación pesquera pone en peligro los océanos y “en los últimos cuarenta años un tercio de la pesca se ha agotado y el 90 por ciento de los grandes predadores marinos, como atunes, tiburones o meros, han sido eliminados. Si la actividad pesquera y la acción 83 La crisis en los océanos y el declive de la pesca mundial demuestra que los tipos más graves de fracaso ecológico surgen de las características de nuestro sistema dominante de racionalidad y de su exceso de confianza y no de la falta de aplicación o extensión de los sistemas de gestión natural, como suele sugerirse. La mayoría de las industrias pesqueras del mundo han sido administradas de forma racional y científica en la mayor parte del siglo XX, pero todas ellas han sido explotadas más allá de los límites sostenibles. Por lo general no se trata de que hayan sido teóricamente reconocidos y objetivamente subestimados los límites ecológicos, tales límites sencillamente no fueron reconocidos ni investigados184. De una forma general, las nuevas tecnologías están propagando enfermedades y contaminación entre los peces locales que son continuamente utilizados como alimento para los peces de piscifactoría, lo que empeora la situación de las poblaciones de peces autóctonos conduciéndolos a la extinción (o extinguiéndoles de hecho, una vez que se utiliza cerca de 4 kilos de los peces locales para llevar 1 kilo de pescado de la factoría al mercado)185. Esto es sólo uno ejemplo de los muchos fallos de la racionalidad ecológica186. Parece que estamos tratando aquí de una inherente inhabilidad en reconocer los límites, restricciones y formas de dependencia de lo que queda fuera de uno mismo. El fracaso en reconocer los límites de otros seres y sistemas es el producto de una visión monológica y profundamente centrada en los humanos. En la misma línea, podemos constatar que la gran falacia de la revolución verde no solo no fue capaz de, siquiera, disminuir el hambre en el mundo, sino que demostró ser más que nada una estrategia de mercado. Menos de 10 grandes corporaciones detentan el comercio global de las principales semillas del planeta. Este nuevo tipo de agricultura “reemplaza los humana continúan ese curso, en 2049 el cien por ciento de la pesca en todos los océanos se "habrá colapsado". Con el objetivo de denunciar esta situación y de buscar soluciones alternativas, el denominado "Green Fórum" ha reunido en Miami a diversos científicos y expertos medioambientales”. Estos son algunos de los mensajes que llegan desde el evento, que ha contado con la colaboración de Televisión Española (28/08/2009); http://www.ecologistasenaccion.org/article20624.html, en 05/09/2012. 184 “La mayoría de los daños ecológicos fueron planeados y llevados a cabo por las universidades altamente cualificadas del occidente, generalmente, bien intencionadas pero educadas, precisamente, en este tipo de racionalidad.”, ORR, DAVID. Ecological Literacy: Education and the Transition to a Postmodern World. 1992, State University of New York Press, Albany, in PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 25. 185 WOOD, C. Our Dying Oceans, 1998, Macleans Magazine, October, 5, pp. 50-59, y PLUMWOOD, VAL. SealsKin, 1992, Meanjin, 51 (1), pp. 45-58, en PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 26. 186 TILMAN, D. y COHEN, J. E. Biosphere 2 and Biodiversity: the Lessons so far, 1996, Science, pp. 11501151; SHIVA, V. y MIES, M. The seeds and the Earth, 1994, (eds) Close to Home: Women Reconnect Ecology, Health and Development, Earthscan, London; STEINBRECHER, R.A. y MOONEY, P. R. “Terminator Technology: the Threat to World Food Security”, 1998. The Ecologist, 28 (5), pp. 276-279, en PLUMWOOD, VAL. Environmental Culture: The Ecological Crisis of Reason, op. cit., p. 26. 84 recursos renovables de la granja, por insumos no renovables de las fabricas” y desplaza el trabajo de las poblaciones locales, sobre todo el de las mujeres, por el de maquinas que producen peligrosos productos agroquímicos, “aumentando la eficiencia del mercado y las ganancias a través de la fragmentación, pero a costa del capital de la naturaleza en suelos fértiles y vivientes”187. Para Vandana Shiva el hecho de que, actualmente, haya en el Tercer Mundo cada vez más pobres padeciendo hambre esta íntimamente relacionado con el modelo patriarcal de progreso que toma las ventas y las ganancias como indicadores de bienestar. El gran problema aquí es que la visión orgánica nutridora de la agricultura, originalmente basada en principios femeninos de cuidado y creatividad fértil (y comparada con la esfera femenina, como ya se ha discutido anteriormente), fue suplantada por una visión masculina de generar ganancias y, al hacer esto, se origina la exclusión de centenas de millares de mujeres, niñas/ niños y pobres del derecho a la alimentación. En este modelo patriarcal, la fragmentación de los procesos separa la silvicultura de la agricultura, ambas de la ganadería y reduce los múltiples productos del bosque a un único producto que es la madera para el uso comercial, olvidándose que de los animales también se obtiene abono y energía y reduciendo éstos a carne, leche y otros recursos. Vandana Shiva denuncia que en pocos años tras la revolución verde se suplantaron cuarenta siglos de conocimiento agrícola que fueron borrados por corporaciones transnacionales y por los técnicos varones del occidente, que homogeneizaron la diversidad de la naturaleza y del conocimiento humano en un modelo reduccionista de agricultura, cambiando las formas orgánicas de vivir y basadas en una economía de la naturaleza y de subsistencia, hacia una economía de mercado y creando la especificidad de las semillas híbridas, los fertilizantes químicos, los plaguicidas, la mercantilización y los grandes regadíos188. VS menciona algunos estudios realizados que impugnan la creencia general de que la agricultura tradicional es primitiva y atrasada189. 187 SHIVA,VANDANA. Abrazar la vida: mujer, ecología y supervivencia, 2004, op. cit., p. 153; MIES, MARÍA. y SHIVA, VANDANA. Ecofeminismo. Teoría Crítica y Perspectivas, op. cit., pp. 48-53. 188 SHIVA,VANDANA. Abrazar la vida: mujer, ecología y supervivencia, 2004, op. cit., pp. 153, 155, 157. 189 Uno de ellos enumera las características de los cultivos tradicionales que eran atribuidos a las mujeres, que utilizaban las cenizas como fertilizante, que crearon numerosas herramientas como la hoz, el pico, la pala y el arado simple, el barbecho y la rotación de cultivos, el aprovechamiento de los desechos orgánicos como abono, el cultivo en terrazas, las plantaciones a nivel, el riego y la recuperación de la tierra mediante la plantación de árboles y, además, que los ochos cereales más importantes (trigo, arroz, maíz, cebada, avena, sorgo, mijo y centeno) todos fueron aclimatados por mujeres. Estos estudios comprueban que estas prácticas agrícolas son tan permanentes y perfectas que en ninguna otra parte se encontrarían mejores ejemplos de conservación escrupulosa de la tierras sin malezas y sobre la ingeniosidad de los dispositivos de conservación del agua, del conocimiento del suelo, así como del momento exacto de la siega y la siembra, la rotación y el sistema de cultivos mixtos, etc. 85 También Vandana Shiva, en su largo estudio, desmonta algunos de los mitos creados por las grandes corporaciones de la revolución verde190, sobretodo el de las “semillas milagrosas”, que fueron privatizadas y comercializadas, quitando a las/os campesinas/os del tercer mundo el dominio de los recursos genéticos vegetales. Las semillas, que son el primer eslabón de la cadena alimenticia –afirma VS- fueron producidas, almacenadas y replantadas por la gente de los campos durante más de cinco mil años, permitiendo que la naturaleza siguiera su curso191. La preservación de la diversidad genética y la posibilidad de autorrenovación de los cultivos comestibles siempre han estado asociados al control del germoplasma, que es la fuente de toda la riqueza vegetal, y fue realizada, desde siempre, por las mujeres y campesinas/os del tercer mundo. Con la revolución verde, las semillas –ahora “milagrosas”– también denominadas como variedades de alto rendimiento, pasaron a controlar el mercado de la agricultura, pero, en realidad, no son de alto rendimiento, y lo que las caracteriza es que responden muy bien a elevados niveles de riego y fertilizantes químicos, pues si no cuentan con los insumos ideales, su rendimiento es bajísimo192. Teniendo en cuenta que requieren más insumos, estas semillas desperdician recursos y son muy vulnerables a plagas y a enfermedades. Para VS La revolución verde ha consistido en lograr cultivos que impresionan por su uniformidad genética y su vulnerabilidad. Además de todo esto, la baja resistencia a las sequías y plagas ha provocado graves pérdidas de cultivos193. Estos valores están en proceso de ser rescatados por el movimiento de la Permacultura, como veremos más adelante. Más detalles en: STANLEY, A. “Daughters of Isis, Daughters of Demeter: When Women Sowed and Reaped” in J. Rothschild (ed.), Women, Technology and Innovation, New York: Pergamon, 1982; HOWARD, A. An Agricultural Testament, London: Oxford University Press, 1940; VOELKER, JA. Report on the improvement of Indian, culture, London: Eyre and Spottiswode, 1893, p-11; SHIVA, VANDANA. Abrazar la vida: mujer, ecología y supervivencia, 2004, op. cit., pp. 158-9. 190 También sobre el tema: PEARSE, ANDREW. Seeds of Plenty, Seeds of Want: Social and economic implications of the Green Revolution, UNRISD and Clarendon Press, Oxford, 1980; DASGUPTA, BIPLAB. The New Agrarian Technology and India, UNRISD and Macmillan, London, 1977; PALMER, I. The New Rice in Indonesia, UNRISD, 1977; Some Social and Economic Aspects of the Introduction of New Varieties of Paddy in Malaysia: A village case study, U.N. Bhati, UNRISD, 1976; ALCÁNTARA, CYNTHIA HEWITT de. Modernizing Mexican Agriculture: Socioeconomic implications of technological change, 1940-1970, UNRISD, 1976; The Social and Economic Implication of Large-Scale Introduction of New Varieties of Foodgrain: Summary of conclusions of a global research project, UNRISD, 1974; The Political Economy of Agrarian Change: An essay on the Green Revolution, UNRISD and Macmillan, London, 1974; V.K.R.V. Rao, Growth with Justice in Asian Agriculture: An exercise in policy formulation, UNRISD, 1974; GRIFFIN, KEITH. The Green Revolution: An economic analysis, UNRISD, 1972; PALMER, I. Science and Agricultural Production, UNRISD, 1972; PALMER. I. Food and the New Agricultural Technology, UNRISD, 1972. 191 Aunque tampoco cabe dudar, en este punto, de la dependencia de la humanidad de las condiciones naturales y las cíclicas crisis de hambre que la asolaban periódicamente, hay que considerar que las autoras aquí analizadas, Val Plumwood y Vandana Shiva, no son contra el desarrollo, sino al que consideran mal desarrollo (VS) y/o a la apropiación de la razón y del desarrollo por el poder (VP), que han convertido cuestiones esenciales de supervivencia en variables económicas.	5,663
https://openalex.org/W2744158200	OpenAlex	Open Science	CC-By	2,017	A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing	Lawrence Tinsley	English	Spoken	7,415	11,428	A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing Lawrence Tinsley a,, Christine Chalk a, John Nicholls a, J€orn Mehnen b, Rajkumar Roy a a Through-life Engineering Services Centre, Cranﬁeld University, MK43 0AL, UK b Dept. of Design, Manufacture and Engineering Management, University of Strathclyde, 75 Montrose St, Glasgow, Scotland, G1 1XJ, UK Lawrence Tinsley , Christine Chalk , John Nicholls , Jorn Mehnen , Rajkumar a Through-life Engineering Services Centre, Cranﬁeld University, MK43 0AL, UK b Dept. of Design, Manufacture and Engineering Management, University of Strathclyde, 75 Montrose St, Glasgow, Scotland, G1 1XJ, UK a Through-life Engineering Services Centre, Cranﬁeld University, MK43 0AL, UK b Dept. of Design, Manufacture and Engineering Management, University of Strathclyde, 75 Montrose St, Glasgow, Scotland, G1 1XJ, UK Open Access funded by Engineering and Physical Sciences Research Council. Under a Creative Commons license CC BY 4.0 http://dx.doi.org/10.1016/j.ndteint.2017.08.001 Received 7 November 2016; Received in revised form 31 July 2017; Accepted 1 August 2017 Available online 3 August 2017 0963-8695/© 2017 Published by Elsevier Ltd. A R T I C L E I N F O Keywords: Pulse thermography Thermal Barrier Coating Thermal diffusivity Oxidation ageing This paper presents an assessment of ageing for thin Thermal Barrier Coatings (TBC) using active thermography. As TBCs undergo ageing during their service life, sintering changes the porosity, elements migrate from the substrate, and micro-cracks build up in the structure of the material, exhibiting a change in thermal conductivity and diffusion properties. As the material ages and these properties change over time, it is possible to exploit trends in this change for characterisation of coating ageing, which would provide a diagnostics tool to estimate remaining useful life. In this study, through-depth diffusivity measurement has been applied to thin EB-PVD coatings which are artiﬁcially aged via oxidation furnace cycles. In order to address the difﬁculties of capturing a fast thermal event in a thin coating, a novel parametric study approach has been carried out to optimise data capture and analysis, maximising available frames for the model ﬁtting step. Through-depth dif- fusivities have been measured during ageing for six samples, yielding a repeatable trend in thermal diffusivity measurements, with three features, which can be exploited for ageing characterisation of thin EB-PVD TBCs, and used as an alarm of imminent failure. with a fast thermal event, and to identify and test exploitable trends in the relationship between the thermophysical property of thin EB-PVD TBCs and age, as measured by single-sided thermography, to qualify a test for measuring remaining life of TBCs. 1. Introduction The performance and durability of gas turbines have been signiﬁ- cantly increased with the application of Thermal Barrier Coatings (TBCs), which help protect components from the intense heat from combustion gases. The coatings reduce the temperature of the surface of the allowing higher temperature operation with improved efﬁciency. Contents lists available at ScienceDirect Contents lists available at ScienceDirect Corresponding author. E-mail address: l.tinsley@cranﬁeld.ac.uk (L. Tinsley). 2. Background Over the last decade there has been considerable research in the application of thermography to the non-destructive testing (NDT) of TBC. This has primarily seen application in the detection of delaminations, monitoring of residual stresses at the interface boundary, and more recently in the assessment of ageing through measurement of thermo- physical property with a particular focus on thick air plasma spray (APS) coatings. The aim of this study is to demonstrate the applicability of the thermographic technique to thin Electron Beam Physical Vapour Deposition (EB-PVD) coatings, and explore the difﬁculties with its practical application. In order to address these difﬁculties, various data capture parameters need to be explored in a parametric study, as well as analysis parameters to maximise frame inclusion without ﬂash afterglow or back wall effects. NDT and E International 92 (2017) 67–74 NDT and E International 92 (2017) 67–74 2.1. Thermal Barrier Coatings, sintering and spallation TBCs are coatings of insulating materials used in aero engine and gas turbines, made from a ceramic, which function to reduce the temperature from the surrounding gases that the coated component is subjected to by up to 50–150 C [1]. These are typically applied by air plasma spray (APS) or electron beam physical vapour deposition (EB-PVD), the structure of which is shown in Fig. 1. During the component service life, the components are placed under a variety of conditions which cause ageing to the materials. These factors include high stresses both thermal and mechanical; vibration, and contamination. The usual failure mech- anism of TBCs is related to oxidation-driven spallation at the interface either between the oxide scale layer, known as Thermally Grown Oxide (TGO), and the bond coat, or between the TGO and the ceramic topcoat, It is the aim of this experiment to explore and overcome difﬁculties e aim of this experiment to explore and overcome difﬁcu NDT and E International 92 (2017) 67–74 L. Tinsley et al. Fig. 1. Structure of a typical EB-PVD TBC. Fig. 1. Structure of a typical EB-PVD TBC. applied, with a 1-dimensional thermal model ﬁtted to the data, allowing estimation of the surface coating diffusivity. The samples were exposed to furnace oxidation on a basis of time at temperature. These were artiﬁcially aged over time at over 1100 C for up to 128 h in 16 h intervals, as this provides 8 stages for testing during the coating life in addition to the initial state. Pulsed thermographic data was taken at each interval of the experiment in order to capture changes in thermal diffusivity of the TBC. The thermographic system used for heat application and camera-ﬂash synchronisation was Thermoscope®II by Thermal Wave Imaging Inc. [11]. The infrared radiometer used alongside this system for the data capture was a FLIR SC7600 series model. The radiometer hosts an Indium Antimonide sensor which possesses a full spatial resolution of 640 512 pixels, in the spectral range of 3–5.1 mm in the Mid Wave Infrared spectrum, and noise equivalent temperature difference of 20 mK. 2.1. Thermal Barrier Coatings, sintering and spallation The data capture setup involved the placing of the sample on a ﬁxture which was located in front of an internally reﬂective ﬂash hood containing two Xenon ﬂash bulbs which deliver a short light burst of approximately 2 kJ and 4 ms in length, with the radiometer facing the sample through an open observation window at a distance of 230 mm from the sample. This setup is shown in Fig. 2. Fig. 1. Structure of a typical EB-PVD TBC. where crack and spallation in the oxide layer between the bond coat and ceramic top coat can grow until failure of the coating. A mechanistic approach has been developed to improve the prediction of TBC life in this regard, with particular focus on the physics of stresses driving the nucleation of cracking at the TGO level, which requires knowledge of the TBC's morphology and service conditions endured [2]. A life prediction method has been developed by Beck et al. [3] based on TGO crack growth kinetics, taking into account minimum and maximum temperatures of thermal cycling, as well as dwell time at maximum temperature, however this approach provides a prognostic method, and is not useful for in-service diagnostics. One of the advantages relating to data capture with the Thermoscope system, is that it synchronises frame captures with the triggering of the ﬂash system, which helps overcome long-standing issues in pulse ther- mography about uncertainty of the timing of the start of the ﬂash pulse in relation to the surrounding frames. However, in order to accurately obtain a measure of diffusivity from the data, it is important to cut the correct segment of the data sequence for the model to ﬁt. Firstly, an offset is required to initiate the data sequence after the ﬂash and its immediate afterglow, and second is the requirement for a frame “window” limit, so that the later frames in the sequence fed to the model ﬁtting do not contain a second inﬂection point where the heat pulse reaches the back wall of the substrate. This window is shown in Fig. 3. Other research by Pint et al. 3.2. Samples Experiments were performed on six samples of identical design. The Experiments were performed on six samples of identical design. The Fig. 2. Thermographic data capture experimental setup. 3.1. Two-layer thermal model A one-dimensional model for the cooling of a uniformly heated, semi- inﬁnite 2-layer system is ﬁtted to the measured temperature evolution over time (1). This is ﬁtted by a non-linear best ﬁt, returning an esti- mation of diffusivity, effusivity, thickness and coefﬁcient of reﬂection, depending on what is known and what is desired [12]. Meanwhile, the life prediction methodology by Busso et al. [2] pro- vides a method to facilitate in-situ life inspections. However, it also has challenges in requiring various inputs to be known, including full description of the TBC and its interface roughness, including the thermal cycling that the part has been subjected to. TðtÞ ¼ Q0 εc ﬃﬃﬃπ p t 2 641 þ 2 X ∞ n¼1 Γne n2l2c αc t 3 75 (1) (1) 2.1. Thermal Barrier Coatings, sintering and spallation [4] takes a different approach and studies migration of elements between the substrate and the TGO, and the resulting stresses in the layer, where Aluminium can diffuse and lead to the formation of other oxides, and Nickel may be more susceptible to the formation of voids, leading to spallation at the interface. Other elements in the alloy can migrate more easily. Between these different mecha- nisms, a bond coat may function in allowing oxide scale formation, but may also be more prone to formation of voids. There are multiple drivers behind TBC failure relating to the structure of coating, the material used, its thermal cycling, minimum and maximum temperature, and mechan- ical loading. These developments are useful in providing an estimate of TBC life on various designs and the dominant factors in well-known service conditions, however, it would be challenging to apply their approach in a service context. 2.2. Diffusivity-based ageing characterisation A variety of research has been conducted on the topic of categorising the remaining useful life of TBCs. There is a requirement in industry to evaluate TBC ageing in-situ during their service life to predict remaining useful life and improve decisions at maintenance intervals. Measurement of stresses or change in material properties can facilitate this need. where ε ¼ ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ kρCp p is the effusivity in W/cm2/K/s0.5, Γ ¼ (εcεs)/(εcþεs) is the reﬂection coefﬁcient between the coating (c) and substrate (s), and lc refers to coating thickness. αc is coating diffusivity in m2/s, t is the time after the ﬂash pulse, in seconds. Bison, Cernuschi, Grinzato, Marinetti & Robba [5] developed a thermographic inspection technique to estimate in-plane and in-depth thermal diffusivity measurements of TBCs, which was applied for through-thickness measurement of APS TBCs, anticipating no difference between the directions. This was later applied to measurement of both in-plane and through-thickness of APS TBCs [6], and then to APS TBCs that were subjected to furnace oxidation cycling [7,8]. As these studies were targeting a niche area of NDT not covered by other techniques, such as Photo Luminescence Piezo Spectroscopy [9], they focused on APS TBCs between 300 and 400 μm in thickness, leaving thin EB-PVD TBCs less than 200 μm thick to other techniques. 3.3.2. Design of experiments The 5 key factors were used as inputs. The trend sought was a stable point in the output of thermal diffusivity estimation and a high coefﬁ- cient of determination, R2. From the equipment used, a full-factorial experiment with all possible levels included would be very large. Thou- sands of levels of integration times are available in custom speciﬁcation, while there are several ﬂash power options available in the Thermoscope II™system. Initially limiting (a) integration time to 5 levels – the cali- brated default, surrounded by four 1000 μs steps spread either side – (b) the ﬂash power to four wide steps, (c) application of soot coating on/off, (d) 5 frame offsets after the ﬂash of 0–4 and (e) window size from 10 to 40 frames (31 levels), initially reduces the number of experiments to a partial factorial of 40 experiments per sample, and 6200 outputs, factoring all analysis parameters (d) and (e). This can be reduced further by limiting the number of window limit parameters to a handful of steps spread within 10–40 frames. The steps chosen were ﬁner within the ﬁrst 10 frames to capture initial variation of the output, which should begin reaching stability beyond 20 frames as both sides of the interface boundary come to be held in the data window, and as extra steps are added in steps of 10 frames (50 ms) beyond that, it is expected for the output to shift from that stable region. With 0 and 1 frame offset, starting the data window at the ﬂash frame timestamped by the Mosaiq system, and 5 ms after the ﬂash: these are expected to vary their output mea- surements signiﬁcantly, as the ﬂash duration and afterglow will domi- nate the ﬁtting of eq. (1). The partial factorial setup reﬂecting this parameter reduction is shown in Table 1. Integration time is the time duration that the pixels on the detector chip are exposed to incident ra- diation before refresh and data readout. An issue with performing high framerate inspection is that in order to obtain a high framerate, the maximum framerate is limited by the integration time used; the longer the integration time, the lower the maximum framerate available. In all experiments, the framerate of 200 Hz was used, as this was the maximum achievable framerate of the longest integration time, and would be available to all experiments, eliminating it as a factor. 3.3. Parametric study A key issue of the experiment involves the wide range of parameters that affect the temperature readings feeding into the diffusivity estima- tion. The full number of factors could be extensive, depending on the available equipment. A laser heat source can have a more carefully controlled heat duration than ﬂash lamps, and a shorter pulse, which in the ideal case modelled in equation (1) would require a higher energy or a more intense beam to facilitate high energy input in a shorter burst. In literature, the setup description of an experiment is usually sufﬁcient to allow an approximate replication; however, some factors, such as inte- gration time of the detector, and the parameters of the exact data window within the full data capture, are not usually disclosed, resulting in some ambiguity which would usually be ﬁlled with direct information requests or educated guesses on which parameters would be ideal. Even then, a narrow selection of parameters without comparison is likely, and there is an uncertainty on the settings of the inputs, and how the resulting output may compare with parallel experiments with parametric variation. From the equipment available, the experiment entailed 5 key factors, each with varying number of levels available for selection with each, which are: (a) integration time of the infrared detector, which is the equivalent of exposure time for a photographic camera, in this case referring to the exposure time before read-out of a shutter free detector, (b) the ﬂash power intensity, which in this case is a % of maximum power intensity, referring to the % of the capacitor bank's full output used for the ﬂash pulse, (c) the application of a soot coating to the surface to improve emissivity, which is usually applied in all experiments consid- ering the translucency of TBCs in infrared and visible light [13,14], however it is desirable for real-world tests to forego this requirement, and (d) the beginning and (e) end of the data window that is used for the calculation. The last two factors are important at the analysis stage because of difﬁculty in determining the termination of the ﬂash and its afterglow, as well as the need with a fast thermal event to use a sufﬁcient number of frames for calculation. 3.3.2. Design of experiments In order to achieve this framerate, ¼ windowing mode was used. Longer integration times would reduce the maximum framerate, while lower framerates would reduce the amount of data available to ﬁt the model to. samples were made with a platinum diffused bond coat, the ceramic was Zirconia-7wt. % Yttria topcoat deposited via Electron Beam Physical Vapour Deposition, of uniform thickness of approximately 150 μm. The substrate used was CMSX-4. 3. Experimental setup Thermophysical material properties are typically measured using the Flash Method [10]. In this experiment, the front-face ﬂash method is Fig. 2. Thermographic data capture experimental setup. 68 Fig. 3. Illustration of the data output showing different regions within the data, from pre- ﬂash (a), ﬂash duration and post-ﬂash heating effects (b), coating data window (c) and heat pulse diffusion into the substrate material (d). L. Tinsley et al. NDT and E International 92 (2017) 67–74 L. Tinsley et al. beginning of the coating's thermal diffusion is uncertain. The window limit introduces an uncertainty where it is logically sound to select an arbitrary point beyond the heat pulse encountering and crossing into the substrate. However, owing to the model-ﬁtting methodology, or the polynomial ﬁtting of the TSR peak second differential method [15], it is important to have abundant data covering both sides of the transition while not using frames affected by the termination of the substrate. Usually for thick coatings and large substrates with slow thermal events, making an educated selection based on calculations is sufﬁcient. How- ever, with thin coating inspection, while the substrate is much thicker in comparison, in terms of diffusion time through a highly conductive metal, both layers experience a fast thermal event for obtainable fram- erates. Seeking the optimum data window for calculating the diffusivity is a key exercise. With only a few dozen frames within the diffusion time available at accessible framerates, any initial frame still in the ﬂash pulse will result in under ﬁtting the solution to the data. A low number of frames beyond the coating-substrate interface will increase the effect of noise and result in a poor ﬁt, while excess frames beyond the interface would encounter the inﬂuence of the rear surface of the substrate. In order to identify the correct data window, as well as setup parameters, a design of experiments methodology was adopted. Fig. 3. Illustration of the data output showing different regions within the data, from pre- ﬂash (a), ﬂash duration and post-ﬂash heating effects (b), coating data window (c) and heat pulse diffusion into the substrate material (d). 3.3. Parametric study factors, a handful of ‘soft’ factors, such as the effects of different waiting time between captures, to offset heat build-up within the ﬂash hood; and data captures at different times of day with changes in ambient tem- perature and humidity in the lab environment. The soft factors tested produced no measurable effects. 3.4. Furnace cycling TBCs Having condensed the number of factors, the samples were inspected at each stage of the experiment from initial pre-exposure to every stage between the 16 h cycles, until the onset of coating spallation. At each stage, samples were removed from the furnace, left to cool for 15 min, at which point they had cooled to approx. 60 C, and were quenched in distilled water. The samples were subjected to a thermographic inspec- tion without then with a graphite soot coating applied to increase emissivity of the surface. This soot coating was partially removed with water in an ultrasonic bath then isopropanol in an ultrasonic bath, and returned to the furnace. Any soot residue would be vaporised in the furnace, leaving the samples clean and uncontaminated for the next soot- free inspection stage. The full cycle of this process is shown in Fig. 4. This allows selection of optimum analysis parameter combinations that result in stable regions of results, seen here in the center set (circled), indicating ﬁrst that offset is a very strongly inﬂuential factor, showing strong variations from left (offset of 0) to right (4); with window limit also having a strong inﬂuence, within each offset subset. As the estima- tions for offset of 2 indicate, the inﬂuence of window limit is signiﬁcantly reduced with the optimum offset value, achieving a stable region where R2 coefﬁcient is high, and diffusivity measurement has a low variation within the offset setting. This was consistently found not to be the case with all other offset values. This also held further to experiments with variations of the other factors, giving a full variation in the outputs, shown in Fig. 6. In the ﬁgure, many experiments were performed with 4. Results 4.1. Parametric study 3.3. Parametric study The resulting inputs and levels would produce 40 physical experi- ments, with 35 analysis variations on each, yielding 1400 outputs for a single set of tests for one sample at each point in the experiment. The partial factorial experiment design was run for a single sample before the furnace cycling stage to reduce the total number of experiments. In the initial stage, 54 experiments were conducted, which included variations of parameters and their levels in Table 1. In addition to the identiﬁed 3.3.1. Data window For ﬂash lamp heat sources, the termination of the heat pulse and For ﬂash lamp heat sources, the termination of the heat pulse and 69 NDT and E International 92 (2017) 67–74 L. Tinsley et al. L. Tinsley et al. Table 1 Design of experiments matrix for parametric study – gaps indicate no additional levels selected, as different factors have different levels available. Index Factor Units Level (1) Level (2) Level (3) Level (4) Level (5) Level (6) Level (7) A Integration time μs 1000 2000 2271 3000 4000 – – B Flash intensity % max power 25 50 75 100 – – – C Soot coating applied No Yes – – – – – D Frame offset number 0 1 2 3 4 – – E Window limit number 10 12 14 16 20 30 40 a single data capture is stretched over its 35 analysis iterations, with every set of several data points representing each of ﬁve offset values between 0 and 4; and within each offset subset are the window limits from Table 1. The full results of this parametric study are shown in Fig. 6, which displays the output estimation of thermal diffusivity against the partial factorial parameters, as well as other variations discussed above. The objective of this exercise being to seek out parameter combinations which achieve stable regions of diffusivity estimation without signiﬁcant variation or noise. As the parameters approach optimal settings, the output stabilises, giving an optimal solution where the outputs show little variability with adjustment of factor levels, as seen in Fig. 5. In the ﬁgure, offset and window limit parameters are varied along the x-axis, where every several points along the axis are the window variations between 10 and 40, within each set for offset parameter, 0–4, from left to right. 4.1. Parametric study 4.1. Parametric study A single experiment of the parametric study is shown in Fig. 5, where A single experiment of the parametric study is shown in Fig. 5, where Fig. 4. Flowchart of inspection process during the furnace cycle. The process repeats until the TBC spalls in either of the last two steps of stage (iii). 70 rt of inspection process during the furnace cycle. The process repeats until the TBC spalls in either of the last two steps of stage (iii). 70 NDT and E International 92 (2017) 67–74 L. Tinsley et al. Fig. 5. Comparison of analysis parameters (x-axis) against their resulting diffusivity measurement in a single data capture at one integration time and ﬂash power. Fig. 5. Comparison of analysis parameters (x-axis) against their resulting diffusivity measurement in a single data capture at one integration time and ﬂash power. data provided ﬁts the ideal model. This can indicate that a strong model ﬁt is possible with temperature data from different integration times, as shown in Fig. 7, which also indicates a weaker ﬁt generally for offset values of 0 and 1 (x-axis values 1–7, and 8–14 respectively). While integration time shows a strong variation in R2 between levels of 1000 and the other levels, their output diffusivity values shown in Fig. 6 – y- axis 50-54 respectively – shows a strong output correlation between integration times of 1000–2271. Integration time is an important factor since shorter values are required for faster framerates, while longer integration times are required for lower temperatures. A low integration time at a given temperature may achieve a low saturation of the sensor. The range of saturation recommended by the manufacturer is 20–80%. From the manufacturer's software, an integration time of 1000 μs, for a surface emissivity of 0.9 yields 5.3% saturation at 20 C, and 7.5% at 30 C. This is the range of sample surface temperatures during the post- ﬂash heat diffusion event. Meanwhile at the same range, 4000 μs pro- vides 21.1% and 30.0% respectively. Objectively, the longest exposure from the set is the only one that meets the recommendations, however, as can be seen in Fig. 7, it produces the worst performance in matching ideal behaviour. 4.1. Parametric study Here is shown the R2 values for 5 experiments at 50% ﬂash power over different integration times. Fig. 7. Searching for stable settings using R2 indicating accuracy of ﬁt of the two-layer model to the data under the given experimental conditions. Here is shown the R2 values for 5 experiments at 50% ﬂash power over different integration times Fig. 7. Searching for stable settings using R2 indicating accuracy of ﬁt of the two-layer model to the data under the given experimental conditions. Here is shown the R2 values for 5 experiments at 50% ﬂash power over different integration times. R2 indicating accuracy of ﬁt of the two-layer model to the data under the given experimental conditions. Here is shown the R2 values for 5 rent integration times. Fig. 7. Searching for stable settings using R2 indicating accuracy of ﬁt of the two-layer model to the data under the given experimenta experiments at 50% ﬂash power over different integration times. from 25 to 100%. In Fig. 8, it can be observed that lower ﬂash powers attain a higher R2 parameter. However, it can also be observed with experiments 1, 14 and 26 of Fig. 6, where ﬂash power of 25% is used, diffusivity is noisier and less stable, when compared to the following ten experiments in each case, where the other 3 ﬂash power levels are used. There are some notable differences between the inspections with and without soot coating. First of all, the tests without soot application exhibit a large initial jump that the coated inspections do not. This may be due to contamination of the coating from the initial application of soot exhibiting effects on the reﬂectance or apparent effusivity of the TBC that endures through an ultrasonic wash and the ﬁrst 16 h furnace run. However, it could also be a result of the decrease in translucency TBCs undergo during service. Finally, the application of soot coating is considered, and as seen in Fig. 9, the application of soot makes little difference to the R2 value for frame offsets 2–4, indicating a consistent ﬁt of the model in either case. Additionally, there is some improvement of coefﬁcient with increase in integration time, however this is exclusively in the 0–1 offset region that has already been excluded. 4.1. Parametric study However, for offsets of 2 and greater, with window limits 14 and greater, the integration times yield high R2 ﬁts, with deviations at window limits of 30 and 40 (x-axis 20, 21, 27, 28, 34, and 35), with greater deviation introduced by integration time 1000 μs. Consequently, two integration times of 2271 and 4000 μs were selected for the main experiment, in order to reduce the total number of data captures, while capturing both demonstrable accuracy, and the objectively ideal setting in spite of its minor deviation. Fig. 5. Comparison of analysis parameters (x-axis) against their resulting diffusivity measurement in a single data capture at one integration time and ﬂash power. experimental parameter variations along the y-axis, while each set of several points along the x-axis represent each of the window variations for each level of offset value. The ﬁrst seven points along the x-axis containing data for offset of 0, with frame limits of 10, 12, 14, 16, 20, 30, 40, allowing both classes of parameters to be compared. In this way, the other factors can be interrogated for the variance of their outputs and can also be tested according to the coefﬁcient of determination, R2, related to the ﬁtted model, indicating how close the The next parameter considered was ﬂash power, ranging in four steps The next parameter considered was ﬂash power, ranging in four steps Variation of estimated diffusivity values spread over experimental factors and levels such as integration time and ﬂash power (y-axis) against variation in analysis factors and leve ff t d i d i ( i ) Fig. 6. Variation of estimated diffusivity values spread over experimental factors and levels such as integration time and ﬂash power (y-axis) against variation in analysis factors and levels of frame offset and window size (x-axis). alues spread over experimental factors and levels such as integration time and ﬂash power (y-axis) against variation in analysis factors and levels Fig. 6. Variation of estimated diffusivity values spread over experimental factors and levels such as integration time and ﬂash power (y-axis) against variation in analysis factors and levels of frame offset and window size (x-axis) 71 L. Tinsley et al. NDT and E International 92 (2017) 67–74 Fig. 7. Searching for stable settings using R2 indicating accuracy of ﬁt of the two-layer model to the data under the given experimental conditions. 4.2. TBC ageing The experiment was performed on a set of 6 EB-PVD YSZ TBC sam- ples. All 6 samples were subjected to up to 128 h of oxidation ageing, with coatings failing between 80 and 128 h. For this experiment, only through-the-thickness diffusivity was measured due to the technical limitations of the experimental apparatus. The results are shown in Figs. 10 and 11. As seen above, there is a sharp early drop in diffusivity in the soot-free inspections, which then levels, before increasing and decreasing once more at the end of coating life. However, the inspection with a soot coating applied shown below is completely free from this initial jump in values, while it still exhibits a later decrease after 16 h, and a similar short increase and decrease in diffusivity values at the end of the coating life. 4.1. Parametric study While there are consistent trends apparent over age, these exhibit a different shape, and are not as pronounced as trends identiﬁed by anal- ysis of APS TBCs [8]. This may be related to the morphological structure differences between APS and EB-PVD, which could be investigated with a variation of experimental setup that can measure both the through-thickness and in-plane thermal diffusivities [16]. The experimental factors and their levels for analysis was narrowed down to a limited subset, which are shown in Table 2. All measurements were taken at 200 Hz. In both the soot coated and uncoated inspections, similar features of change are shown: an initial decrease, a gradual rise until 112 h, and a ﬁnal drop before sample failure. Changes to thermal properties can be multi-faceted in their source. Thermal conductivity is inﬂuenced by point defects and grain boundaries of the ceramic [17]. Meanwhile, TBCs subject to sintering lose intra-columnar ﬁne porosity, producing an in- crease in thermal conductivity during exposure at temperature. As the TGO layer grows under exposure, cavitation, rumpling and local inter- facial separations form at the TGO and propagate over time, forming defects and obstructions to heat conduction [18]. Considering these factors, while non-soot coated TBCs undergo a rapid initial drop in apparent diffusivity, this is mirrored to a lesser extent in the soot coated inspections between 16 and 32 h, the cause of which will not simply be related to the semi-transparency of the TBC. With further sintering effects during ageing, an increase in diffusivity is expected, and is likely the source of the rise up to 96–112 h in both soot coated and uncoated in- spections. Finally, a sudden drop in diffusivity prior to failure may coincide with the development of TGO growth, cavitation and interfacial cracks at the end of TBC life, which would shift the data away from the ideal two-layer model used. 5. Discussion Immediate observations from the results are that there is no unique relationship between diffusivity and coating age that would permit a one- point inspection randomly at any point in the sample's life to determine remaining useful life. However, it is noted that a trend is exhibited in diffusivity change that may be exploitable. In the case with soot coated samples the thermal diffusivity decreases, and then experiences an in- crease at around 96 h, before a ﬁnal decrease. The late-life increase is also exhibited in non-soot coated samples. This pattern could be useful to diagnose the coating reaching the last stages of its life. With only a handful of samples surviving into the ﬁnal decrease stage with the 16 h cycles used, the decrease itself could be more useful as an alarm indi- cating imminent failure, than as a feature of an ageing characterisation technique, unless more frequent inspections could be applied at the ﬁnal stages to catch the onset of the terminal decline. One of the main ﬂaws in this experiment is that the unpainted in- spections are affected by the semi-transparency of TBCs, which are particularly prominent in the mid-wave IR band that has been used. This could be improved with use of a long-wave IR radiometer, or application of different ﬁtting models that can accommodate for multiple layers [19], or semitransparency for emission as well as optical excitation [20,21]. In order to exploit these trends in service, these accommodations would need to be made, along with removing the effects of surface dis- colouration [14], while other techniques can be used to exploit the semi-transparent nature of TBCs, such as with mid-wave infrared reﬂectance imaging [22]. Data presented in this article may be accessed via Open Access [23]. 72 Fig. 8. Testing at constant integration time of 2271 μs, are different levels of ﬂash power tested for their R2 stability over different ﬁtting parameters. Fig. 9. R2 difference achieved without soot, compared to with soot, and with soot at the integration time of 4000 μs. This shows that offset of 0 and 1 frame (x-axis 1-14) are particularly erroneous, resulting in poor ﬁts of the model to the data. L. Tinsley et al. NDT and E International 92 (2017) 67–74 NDT and E International 92 (2017) 67–74 L. Tinsley et al. Fig. 8. Table 2 Table 2 Final experimental setup for furnace cycle experiment. Factor Name Units Levels Levels A Integration time μs 1 2271 B Flash intensity % max power 1 50 C Soot coating Yes/No 2 Yes/No D Frame offset number 1 2 E Window limit number 1 20 E Window limit number 1 20 Fig. 10. Normalised diffusivity box plot of all samples over furnace exposure time, inte- gration time 2271 μs, ﬂash power 50%, offset 2, frame limit 20, without soot, showing a pattern between the samples over ageing. Fig. 11. Results for the same parameters, with soot coating, showing a similar late-age rise trend as without soot. The large step down from an initially high diffusivity has apparently disappeared, although a smaller initial decrease is present. 5. Discussion Testing at constant integration time of 2271 μs, are different levels of ﬂash power tested for their R2 stability over different ﬁtting parameters. integration time of 2271 μs, are different levels of ﬂash power tested for their R2 stability over different ﬁtting parameters. ig 9 R2 difference achieved without soot compared to with soot and with soot at the integration time of 4000 μs This shows that offset of 0 and 1 frame (x-axis 1-14) are particular Fig. 9. R2 difference achieved without soot, compared to with soot, and with soot at the integration time of 4000 μs. This shows that offset of 0 and 1 frame (x-axis 1-14) are particularly erroneous, resulting in poor ﬁts of the model to the data. t, compared to with soot, and with soot at the integration time of 4000 μs. This shows that offset of 0 and 1 frame (x-axis 1-14) are particularly del to the data. Fig. 11. Results for the same parameters, with soot coating, showing a similar late-age rise trend as without soot. The large step down from an initially high diffusivity has apparently disappeared, although a smaller initial decrease is present. 6. Conclusions This paper has shown the application of the active therm ﬂash method for determining through-thickness thermal diffusiv thin EB-PVD TBC samples in both the default condition and emissivity-improving soot coating applied. Under the limiting co of performing the ﬂash test for through-thickness measuremen thermal event, a parametric study for setting up the thermogra Table 2 Final experimental setup for furnace cycle experiment. Factor Name Units Levels Levels A Integration time μs 1 2271 B Flash intensity % max power 1 50 C Soot coating Yes/No 2 Yes/No D Frame offset number 1 2 E Window limit number 1 20 Fig. 10. Normalised diffusivity box plot of all samples over furnace exposure time, inte- gration time 2271 μs, ﬂash power 50%, offset 2, frame limit 20, without soot, showing a pattern between the samples over ageing. Fig. 11. Results for the same parameters, with soot coating, showing a simi rise trend as without soot. The large step down from an initially high diff apparently disappeared, although a smaller initial decrease is present. Table 2 Final experimental setup for furnace cycle experiment. 6. Conclusions This paper has shown the application of the active thermographic ﬂash method for determining through-thickness thermal diffusivity of six thin EB-PVD TBC samples in both the default condition and with an emissivity-improving soot coating applied. Under the limiting constraints of performing the ﬂash test for through-thickness measurement of a fast thermal event, a parametric study for setting up the thermographic data Fig. 10. Normalised diffusivity box plot of all samples over furnace exposure time, inte- gration time 2271 μs, ﬂash power 50%, offset 2, frame limit 20, without soot, showing a pattern between the samples over ageing. 73 L. Tinsley et al. NDT and E International 92 (2017) 67–74 capture and analysis has been developed. The monitoring of thin EB-PVD TBCs during oxidation ageing has been undertaken, and has shown that the method is able to discern changes in through-thickness diffusivity over coating life, and that these changes are still observable without soot coating in the non-ideal performance of the test, even while the soot-free estimation will be inaccurate due to semi transparency. It is observed that there is not a unique relationship between through-thickness diffusivity value and ageing alone, where one range of measured values would translate directly into a range of life estimates, however, a history of measurements may be required for the part, in order to classify its age. The observed trends were not as clear as those observed by other re- searchers with APS TBCs, and this may be related to differences in through-thickness and laminar diffusivities, owing to EB-PVD TBCs' differing structure from APS TBCs. The thermal property changes observed provide three features of diffusivity trends that appear both in coated and uncoated inspections, and may be useful for estimation of coating life, while providing a marker that could indicate TBC close to the end of its life. [5] Bison PG, Cernuschi F, Grinzato E, Marinetti S, Robba D. Ageing evaluation of thermal barrier coatings by thermal diffusivity. Infrared Phys Technol 2007;49(3): 286–91. [6] Bison P, Cernuschi F, Capelli S. A thermographic technique for the simultaneous estimation of in-plane and in-depth thermal diffusivities of TBCs. Surf Coat Technol 2011;205(10):3128–33. [7] Cernuschi F, Bison P, Marinetti S, Campagnoli E. Thermal diffusivity measurement by thermographic technique for the non-destructive integrity assessment of TBCs coupons. Surf Coat Technol 2010;205(2):498–505. [8] Cernuschi F, Capelli S, Bison P, Marinetti S, Lorenzoni L, Campagnoli E, et al. 6. Conclusions Non- destructive thermographic monitoring of crack evolution of thermal barrier coating coupons during cyclic oxidation aging. Acta Mater 2011;59(16):6351–61. [9] Christensen RJ, Lipkin DM, Clarke DR. Nondestructive evaluation of the oxidation stresses through thermal barrier coatings using Cr piezospectroscopy. NDT E Int 1998;31(2):130. [10] Parker WJ, Jenkins RJ, Butler CP, Abbott GL. Flash method of determining thermal diffusivity, heat capacity, and thermal conductivity. J Appl Phys 1961;32:1679–84. [11] Thermal Wave Imaging Inc., Thermoscope II: advanced thermography for in-service inspection. [Online]. Available: http://www.thermalwave.com/1/376/ thermoscope_ii.asp. [Accessed 10 January 2016]. [12] Bison PG, Cernuschi F, Grinzato E, Marinetti S, Robba D. Ageing evaluation of thermal barrier coatings by thermal diffusivity. Infrared Phys Technol 2007;49(3): 286–91. [13] Liebert CH. Emittance and absorptance of NASA ceramic thermal barrier coating system. In: NASA technical paper 1190; 1978. [2] Busso EP, Wright L, Evans HE, Mccartney LN, Saunders SRJ, Osgerby S, et al. A physics-based life prediction methodology for thermal barrier coating systems. Acta Mater 2007;55:1491–503. Acknowledgements [14] Ptaszek G, Cawley P, Almond D, Pickering S. Transient thermography testing of unpainted thermal barrier coating (TBC) systems. NDT E Int 2013;59(C):48–56. [15] Shepard SM. “Reconstruction and enhancement of active thermographic image sequences. Opt Eng 2003;42(5):1337. This work has been ﬁnanced by the UK Engineering and Physical Sciences Research Council through the EPSRC Centre in Through-life Engineering Services (grant no: EP/I033246/1) and the centre's in- dustry partners. [16] Bison P, Cernuschi F, Grinzato E. In-depth and in-plane thermal diffusivity measurements of thermal barrier coatings by IR camera: evaluation of ageing. Int J Thermophys 2008;29(6):2149–61. [17] Klemens PG, Gell M. Thermal conductivity of thermal barrier coatings. Mater. Sci Eng A 1998;245:143–9. [1] Boyce MP. An overview of gas turbines. In: Gas turbine engineering handbook. Third. Oxford, UK: Gulf Professional Publishing; 2006. p. 49. [3] Beck T, Trunova O, Herzog R, Singheiser L. TBCs for gas turbines under thermomechanical loadings: failure behaviour and life prediction. In: EPJ web of conferences, vol. 33; 2012. [4] Pint BA, Wright IG, Lee WY, Zhang Y, Prüßner K, Alexander KB. Substrate and bond coat compositions: factors affecting alumina scale adhesion. Mater Sci Eng A 1998; 245(2):201–11. References [18] Clarke DR, Oechsner M, Padture NP. Thermal-barrier coatings for more efﬁcient gas-turbine engines. MRS Bull 2012;37(10):891–8. [19] Akoshima M, Ogawa M, Baba T, Mizuno M. The thermal diffusivity measurement of the two layer ceramics using the laser ﬂash method. J High Temp Soc 2008;34(5): 227–33. [20] Goldammer M, Baumann J. Analytical modeling of ﬂash thermography results for a layered sample. Nondestruct Eval Reliab Micro Nanomater Syst 2002;4703:211–8. y g g y layered sample. Nondestruct Eval Reliab Micro Nanomater Syst 2002;4703:211–8. [21] Abuhamad M, Netzelmann U. Dual-band active thermography on infrared transparent materials. Quant Infrared Thermogr J 2010;7(2):189–200. [21] Abuhamad M, Netzelmann U. Dual-band active thermography on infrared transparent materials. Quant Infrared Thermogr J 2010;7(2):189–200. [3] Beck T, Trunova O, Herzog R, Singheiser L. TBCs for gas turbines under thermomechanical loadings: failure behaviour and life prediction. In: EPJ web of conferences, vol. 33; 2012. [22] Eldridge JI, Spuckler CM, Martin RE. Monitoring delamination progression in thermal barrier coatings by mid-infrared reﬂectance imaging. Int J Appl Ceram Technol 2006;3(2):94–104. [23] Data underlying this study can be accessed through the Cranﬁeld University repository at http://dx.doi.org/10.17862/cranﬁeld.rd.4985051. 74 Cranfield University CERES Research Repository https://dspace.lib.cranfield.ac.uk/ School of Aerospace, Transport and Manufacturing (SATM) Staff publications (SATM) Tinsley L, Chalk C, Nicholls J, et al., (2017) A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing. NDT & E International, Volume 92, December 2017, pp. 67-74 Downloaded from CERES Research Repository, Cranfield University Tinsley L, Chalk C, Nicholls J, et al., (2017) A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing. NDT & E International, Volume 92, December 2017, pp. 67-74 https://doi.org/10.1016/j.ndteint.2017.08.001 Downloaded from CERES Research Repository, Cranfield University https://doi.org/10.1016/j.ndteint.2017.08.001 A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing Tinsley, Lawrence 2017-08-03 Attribution 4.0 International Tinsley, Lawrence 2017-08-03 2017 08 03 Attribution 4.0 International Tinsley L, Chalk C, Nicholls J, et al., (2017) A study of pulsed thermography for life assessment of thin EB-PVD TBCs undergoing oxidation ageing. NDT & E International, Volume 92, December 2017, pp. 67-74	27,846
tel-03833718-DDOC_T_2022_0121_TOUSSAINT.txt_7	French-Science-Pile	Open Science	Various open science	2,022	A contribution to Industry 4.0: a framework to secure standardised data exchange. Automatic. Université de Lorraine, 2022. English. ⟨NNT : 2022LORR0121⟩. ⟨tel-03833718⟩	None	English	Spoken	7,507	15,739	6.4 Conclusion Data manipulation is a cyber-attack aiming to compromise the integrity of data. These attacks can have significant consequences for organisations. Organisations can implement measures to prevent and mitigate data manipulation attacks. These measures help i) identify and protect critical systems and data; ii) monitor networks and detect suspicious activities, and iii) develop responses for mitigating data manipulation events. In this chapter, we propose a Cybersecurity Framework Profile for data manipulation risk management that aligns with the NIST CSF’s security objectives. This Profile aims to help organisations identify and prioritise opportunities to improve their data integrity strategy. In addition to this data manipulation Profile, we propose implementation recommendations to help organisations implement a comprehensive solution for data traceability. These recommendations consist of a collection of user stories for the Subcategories of the Profile, a set of security metrics, and guidelines for information flow awareness training. 129 Industry 4.0 technologies enables organizations to be more connected, which bring new cybersecurity concerns as it creates more entry points that attackers can find and exploit. Organisations are exposed to a growing number of cyber challenges, and as a result, data are also vulnerable to numerous cyber threats, including data manipulation. The growing complexity of the cyber threats led to an increased speed at which tampered data can propagate through organisations and corrupt data exchanges and datadriven decisions. A formal data traceability solution is needed to track data flows across large and complex networks of organisations, and identify and contain the propagation of corrupted data. To address the data traceability challenge, we 1) reviewed existing cybersecurity frameworks to find the framework that would offer a solid and customisable foundation to support our needs for addressing data manipulation risk; 2) proposed a cybersecurity framework for data manipulation risk management, based on the NIST Cybersecurity Framework, to help organisations prevent and mitigate data manipulation events, and finally 3) proposing implementation guidelines (user stories, security metrics, information flow awareness training) to complement the Profile and allowing organisations to develop and implement a comprehensive data traceability solution for data integrity. CONCLUSION AND FUTURE WORK The digital transformation of manufacturing offered by Industry 4.0 has resulted in the generation of large volumes of data. Organisations consume and exchange this data while preserving their integrity and quality. Organisations’ communication and collaboration rely on fast and reliable data exchanges, but these data exchanges come with data interoperability and data traceability challenges. Using information standards to address these challenges, this thesis proposes a new standard-based framework that 1) presents the transition to adaptive project management and a better requirement elicitation model for the standard development process, and 2) provides a formal data traceability solution through a data manipulation Profile to tackle the cybersecurity issues Summary The digital transformation of manufacturing has led to more connected, automated, and data-driven environments and processes. Data has become a key enabler to processes, exchanges, and decision-making. Manufacturing relies heavily on data and the exchange of this data between the different stakeholders, machines, and systems. Data exchange, by definition, is the process of sending and receiving data in a way in which the data content or meaning has not been tampered with during transmission, in other words that the data received is an accurate representation of the data sent. The digitalisation of manufacturing has emphasised the importance of information management, data exchange, and the interoperability of the different actors in the manufacturing processes. The emergence of new technologies and networked data sources support new opportunities for organisational collaboration through high-speed and high-volume data exchange. In other words, this digital era helped improve the speed, volume, accuracy, and consistency of data exchange and innovations across and within organisations. Data is now available and shareable at a quicker rate than organisations can consume and track it. This speed comes with key challenges. In this thesis, we focused on two of these challenges, data interoperability and data traceability, two interdependent challenges that manufacturers face and must understand to address them. On one hand, faster innovation and collaboration are being hindered by data interoperability challenges. Increased collaboration is associated with an increased number of heterogeneous systems that need to communicate with each other. Information standards are a proven solution, as they provide a common language to the information systems that support business and operations in organisations. However, their development process is often long and complex because it requires numerous, international, and heterogeneous stakeholders (e.g., users, technical experts, implementers) to collaborate and reach 131 agreements, oftentimes on a volunteer basis. The standards development process faces inefficiencies that make it incompatible with the fast-paced environment they need to support and provide interoperability for, as well as the ever-evolving nature of the cyber threats that organisations are facing. These inefficiencies therefore slow down standards’ publication and adoption. In this thesis, we propose a transition from predictive to adaptive project management with the use of Agile methods. We present some Agile practices, through a test case using STEP AP242, that can help standard development teams overcome some of these inefficiencies. Agile provides a way (through processes and tools) to 1) improve the management of the available resources (i.e., volunteer staff) by reducing the need for long-term planning and commitment, and 2) improve collaboration between these communities by supporting a proper and faster communication channel. Despite the benefits Agile brings, it also comes with challenges. We identified a clear need for better traceability and visibility of requirements, their elicitation, their management, and their implementation. To address this need, we developed and proposed a new requirement elicitation model that serves as a foundation for providing requirements and decision traceability and visibility, by recording and leveraging project meetings in a formal way. We also proposed an implementation of this model through a customisation of Jira, a tool used by the STEP development teams for bug and issue tracking. On the other hand, Industry 4.0 has enabled manufacturing to become more digitalised, connected, smarter, and more autonomous, leading to a significant increase in the number and complexity of cybersecurity challenges. The digitalisation of Industry 4.0 supports high-volume and high-speed data exchanges across supply chains, enabling a new data-driven approach to business decisions, but also creating a greater attacks surface. These decisions are exposed to the speed at which tampered data can propagate through organisations and corrupt these decisions. Data manipulation can lead to corrupted decisions and processes, as well as compromised entire supply chains, which can have serious consequences for organisations, such as economical and reputational damages. The mean time to identify (MTTI) such a threat is already close to 215 days [17], and the constant growth of data produced and exchanged is likely to push the MTTI upwards. Organisations generally take longer to recover from data manipulation events because 1) unlike ransomware, they take time to be identified, 2) they use data exchange, a key element to industry 4.0, as a propagation vector, and 3) propagation makes it hard to determine the extent and impact of these attacks. While digital signatures have demonstrated their use in identifying such corruption, there is still a need for a formal data traceability framework to track data exchange across large and complex networks of organisations in order to identify and contain the propagation of tampered data. This thesis analysed existing cybersecurity frameworks and their limitations, and proposed a new standard-based framework, in the form of an extended NIST Cybersecurity Framework Profile, to mitigate, manage and track data manipulation attacks. This Profile aims to help organisations align their data integrity prevention and mitigation requirements and resources with the elements of the NIST Cybersecurity Framework, as 132 well as assess the state of their readiness against such attacks. This Profile comes with implementation recommendations to facilitate its adoption and implementation. These implementation recommendations complement the Profile for developing a formal data trace solution to detect and mitigate data manipulation events. These recommendations consist of user stories, security metrics and guidelines for information flow awareness training. Future work As presented in the previous section, we have focused as a priority on some challenges related to data interoperability and data traceability, but there are still challenges to be addressed in order to have the most complete solution possible to ensure fast and reliable data exchange. The framework we propose in this thesis has some limitations. In the next section, we will discuss these limitations and present solutions on how to address them in the future. Data interoperability The Requirement elicitation model we propose in Chapter 4 supports better traceability and visibility of the requirements and decisions. This model is originally designed for the development of information standards, but it can be used by any project with distributed stakeholders. The model presented in this thesis can be extended, that is new attributes (e.g., user story, use case...) can be added to the different concepts, or values of the enumeration lists can be changed, to meet the specific needs of projects. Moreover, this model currently focuses on requirement and decisions gathering and traceability, but it could be extended to support requirement validation and requirement prioritization. For example, a possible enhancement could be to add an additional concept, “Test”, regarding the validation of a requirement or work item. This concept would be intermediate between the Decision and Requirement concepts, and between the Decision and Work Item concepts. This Test concept would be composed of attributes such as acceptance criteria, unit test, integration test, acceptance test or user test for example. This concept would represent the results of the different testing that support the validation of the requirements and/or work items. The next step for this model is to display the information captured in a meaningful way to the various project stakeholders. Visibility is key to Agile management and project information is often shared graphically in an information radiator that is easily accessible and understandable by all stakeholders. Visual representations of requirements and associated work items help communicate, clarify and validate them, by minimising complexity and misconceptions, as well as identifying gaps in requirements. Identifying the most appropriate visualisation techniques to display the information collected through the model would be the first task in this next step. In addition to these visual requirement models, the model 133 should also support report generation. Like the visual representation of the requirements, reports provide visibility. Reports help communicate how effectively the project is progressing as well as highlight the challenges and risks the project is facing. These two functionalities would bring significant benefits to project teams, ensuring that the requirements align with the business needs, that all the stakeholders understand the requirements and their context, and that the project is progressing well. In this thesis, we focus on improving the standards development process to facilitate and promote their adoption and thus address the data interoperability issues. As mentioned, the standards development process is long, and this thesis proposes a solution to shorten the development iterations and increase the delivery velocity of standards development teams, by transitioning to adaptive project management, leveraging Agile methods, and integrating the Requirement elicitation model. The creation and development of a Requirements acceptance framework can also help understand what slows down the standards development process. This framework will use some of the information captured by the Requirement elicitation model. The objective of the Requirement acceptance framework is the analysis of the decision taken during the meetings. This framework will measure and analyse the process of capturing and accepting the requirements by the participants. It will also help project teams to get an overall idea of how the development process is going. Indeed, this framework will evaluate the decisions taken to understand what may cause some disruptions to the project. Finally, a future step would be to develop long-term storage of standard requirements. The standard development iterations currently take between 18 and 48 months, and standards may be developed through multiple iterations. As a result, it is crucial to ensure that any stakeholders of the standard development can access requirements data during the entire standard development process. An ideal solution would be to store both requirements that have been implemented and requirements that have been rejected in order to maintain traceability of the progress of the industry needs. It would also be beneficial to save information about the context of the requirements and the discussions that have happened about the requirements. In addition, this long-term storage solution will need to ensure the integrity of the requirements data over time. Data traceability In this thesis, we focused on data manipulation, which has become a growing threat in cybersecurity. Data manipulation, the concept of subtly modifying data in order to corrupt processes, decisions, or products, can take different forms. As a result, our future work will focus on defining a cybersecurity taxonomy of data manipulation threats. This taxonomy will characterise and classify these threats to allow a precise definition of vulnerabilities that they exploit. Besides, while tools continue to be refined to monitor 134 and detect data manipulation attacks, educating and training employees about these attacks is also important. Adding this cybersecurity taxonomy to the information flow awareness training we defined in a previous section, will help employees anticipate, recognise, and report suspicious activities and thus help organisations avoid or limit the damage of this kind of attack. The data manipulation Profile we propose provides organisations with a voluntary, risk-based approach for improving their cybersecurity posture and reducing risks regarding data manipulation. It could be interesting to complete this Profile with standardised vulnerability and asset management tools, which would allow organisations to have better visibility into the most vulnerable parts of their system. Some components of the SCAP framework we present in Section 5.1.2.4, could be used for this purpose. First, for asset management, one could use: Asset Identification (AID) and Asset Reporting Format (ARF) to uniquely identify assets and thus facilitate reporting and correlation of asset information throughout and between organisations; the Common Platform Enumeration (CPE) entity to automate software inventories and enable security compliance checking; finally, the identification schema Software Identification Tagging (SWID) to track the software installed on the devices belonging to the organisation. Second, regarding vulnerability management, it might be useful to add to our solution: Common Vulnerabilities and Exposures (CVE) to identify security vulnerabilities and correlate vulnerability information about software; the Common Vulnerability Scoring System (CVSS) to estimate the severity of a vulnerability and thus prioritise the organisation's responses to known vulnerabilities. All these are only examples of what could be done in the event that we complete our Profile with SCAP. Finally, the next step will be to develop a tool to implement the features we discussed in Section 6.3.1. This tool will enable organisations to map and monitor the data flows within their supply chains in order to identify risks and vulnerabilities, and thus better protect them. This tool will also enable organisations to catalog their assets (i.e., physical devices, systems, external systems, software, and applications) and inventory the systems of their employees. This tool will also include features to measure and represent the security metrics we defined in Section 6.3.2, as well as features to help create a security training and generate reports based on the recommendations, we made in Section 6.3.3. In addition, this tool should also include the two points discussed previously, that is allowing employees to access the taxonomy on data manipulation threats and incorporating some of SCAP components to help IT teams with vulnerability and asset management. Moreover, to monitor the activities traffic, it would be helpful to define a framework to monitor the flows of HTTP requests, GET and POST, from the different systems as well as the flows of operations (i.e., create, read, update and delete) on the different databases, to determine who is consuming or modifying data. This framework should be integrated into the tool to help with the monitoring of the data flows. Data is a key asset for organisations as it plays a crucial role in organisational decision-making and strategy. Data exchanges need to be accurate and understandable by all organisational systems and stakeholders for decisions and processes based on these exchanges to be reliable and trustworthy. Therefore, in this thesis, we addressed two main and interdependent challenges related to data exchange: data interoperability and data traceability. This thesis focused on facilitating data exchange while preventing data manipulation through the use of information standards. To conclude, this thesis aimed to reduce the time of 1) development and implementation of data interoperability solutions and 2) data traceability operations in response to cyber-attacks that manufacturers are victims of, in order to meet the speed at which data exchanges are now set up and performed. 136 REFERENCES [1] Reinsel, D., Gantz, J., & Rydning, J. (2018). The digitization of the world from edge to core. IDC. https://www.seagate.com/files/www-content/our-story/trends/files/idc-seagate-dataage-whitepaper.pdf [2] Rüßmann, M., Lorenz, M., Gerbert, P., Waldner, M., Justus, J., Engel, P., & Harnisch, M. (2016). Industry 4.0: The Future of Productivity and Growth in Manufacturing Industries. 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https://openalex.org/W3014515778	OpenAlex	Open Science	CC-By	2,020	Thamar University International Dental Conference - TUIDC 1	Mokhtar Saleh Al-anesi	English	Spoken	11,497	22,166	OPEN ACCESS SCIENTIFIC JOURNAL VOLUME 23 N0 01 suppl 02 - 2020 OPEN ACCESS SCIENTIFIC JOURNAL VOLUME 23 N0 01 suppl 02 - 2020 OPEN ACCESS SCIENTIFIC JOURNAL OPEN ACCESS SCIENTIFIC JOURNAL VOLUME 23 N0 01 suppl 02 - 2020 UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUIT A FILHO” Instituto de Ciência e T ecnologia Campus de São José dos Campos October, 16-18, 2019 Faculty of Dentistry - Thamar University Dhamar, Yemen TUIDC 1 Thamar University International Dental Conference PROCEEDINGS October, 16-18, 2019 Faculty of Dentistry - Thamar University Dhamar, Yemen Assist. Prof. Mokhtar Al-anesi Assist. Prof. Mokhtar Al-anesi SCIENTIFIC COMMITTEE Assist. Prof. Gabr Al-Radom (Head) Professor Abdullah Amran Assoc. Prof. Fadhel Alsanabani Assoc. Prof. Mohsen Al-Hamzi Assoc. Prof. Ahmed Madfa Assist. Prof. Mohammed Alsamhari Assist. Prof. Khalid Aldhorae Assist. Prof. Wadhah Al-Hajj Assist. Prof. Yaser Al-Shraee Assist. Prof. Essam Al-Moraissi Assist. Prof. Mohammed Al-khawlani Assist. Prof. Nashwan Al-Tairi Assist. Prof. Abdulrazzaq Al-Maweri Dr. Nasr Al-Qudaimi Dr. Mohammed Nasser Alhajj MEDIA COMMITTEE Dr. Fawaz Alghobari Dr. Abdulghani Al-Basmi Dr. Omar Alfakih Summary Seccion Page Word of the President ----------------------------------------------------------------------------------- 5 Research Article ------------------------------------------------------------------------------------------------ 7 Index - Title ------------------------------------------------------------------------------------------------------ 37 Index - Author --------------------------------------------------------------------------------------------------- 39 Index - Subject ---------------------------------------------------------------------------------------------- 40 Seccion Word of the President TUIDC 1 TUIDC 1 First of all, I would like to extend our thanks and gratitude to the Brazilian Dental Science journal (BDS) that gave us the opportunity through its official scientific newspaper to add a special supplement for our first scientific conference at Thamar University in Yemen. Thamar University International Dental Conference (TUIDC) is our first international scientific conference represents a new breakthrough in the world of scientific research in a country currently suffering conflicts, turmoil and wars a matter that negatively affects the academic research in all Yemeni universities. Despite of that, with a spirit of determination, the academic staff at the Faculty of Dentistry at Thamar university in Yemen insisted to encounter all these challenges, overcome all obstacles and announce holding this conference which is the first of its kind in Yemen. Holding the conference at this time represents and emphasizes the spirit of hope, fortitude and desire for a better future of scientific research in a country that enjoys peace and stability. Our conference was honored by the participation of elite international academic lecturers and researchers in different fields of dentistry although they could not make it in person to deliver their valuable lectures due to war circumstances that prevented them from traveling to Yemen. Therefore, it was adequate to contact them through Skype application. Moreover, a big number of local professors took part in the conference and enriched the three-day scientific sessions with their lectures and researches in all dentistry and oral health specialties. In addition to the scientific programs in the conference, a number of researchers participated with poster presentations and clinical presentations which have been judged by an evaluation committee and granted three prizes for the first positions. During the conference, a number of scientific workshops and courses have been conducted by specialists in different aspects for training doctors on modern techniques in the big world of dentistry. Word of the President Along with the conference, there was an exhibition for dentistry supplies and materials with the participation of top dentistry companies. With all efforts and all possible ways forward, we held this conference and brought it to light with a good image a matter that has established solid foundations for subsequent conferences with more success in the coming years. TUIDC 1 Finally, I would like to thank my colleagues at the Faculty of Dentistry, Thamar University, Yemen who allowed me to write these lines and represent them as the chairman of the conference. I thank all international and local participants for their response and distinguished participation. I also thank all organizing committees who exerted all efforts to make this event a success. Prof. Mokhtar Al-anesi Conference Chairman of Thamar University International Dental Conference TUIDC1 Prof. Mokhtar Al-anesi Conference Chairman of Thamar University International Dental Conference TUIDC1 Prof. Mokhtar Al-anesi Conference Chairman of Thamar University International Dental Conference TUIDC1 TUIDC 1 Research Article Research Article Thamar University International Dental Conference Research Article Research Article Keywords: Cephalometric X-ray; Caucasian; Yemeni Norms. ESTABLISHMENT OF CEPHALOMETRIC NORMS FOR YEMENI ADULTS AND COMPARING IT WITH CAUCASIAN NORMS Al-Yousefi AA, Al-Labani MA email: dramal2010.as@gmail.com Objective: It was necessary to identify what is considered natural and pleasing for Yemenis because of the increasing numbers of Yemeni patients looking for specialized treatment by orthodontists. Therefore the purposes of our study were to establish norms of cephalometric features for Yemeni adults and to compare it with that of Caucasians. Material and Methods: A total of 100 Yemeni students (fifty males, mean age of 23.6 ±2.1 years, and fifty females, mean age of 21.5 ±3.1 years) with normal occlusions, well balanced faces and no history of orthodontic treatment were involved in the study. Five angular and eighteen linear measurements were used for the skeletal, dental and soft tissue analysis. All participant's Lateral cephalometric radiographs were evaluated. The average values and standard deviations for all angles and measurements were determined. The differences for each measurement between Yemenis and Caucasians were calculated using unpaired t–tests. Results: Yemenis subjects had more retrugnathic mandibular positions (P < 0.05), protrusive mandibular incisors (P < 0.01), more protruded lip positions (P < 0.01), deeper mentolabial sulci (P < 0.01) and steeper mandibular planes (P < 0.001) compared to the Caucasians. Yemenis females had larger lower face height than Caucasian females (P < 0.001). Conclusions: This study shows that Yemeni cephalometric norms are different in comparison to Caucasians and provides specific standards for Yemeni patients to be treated according to their special ones. Keywords: Cephalometric X-ray; Caucasian; Yemeni Norms. 7 7 TUIDC 1 Research Article Thamar University International Dental Conference 02i 02 Research Article Research Article Keywords: Silver; Nanoparticles; Toxicity; Mice. TOXICITY ASSESSMENT OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL ADMINISTRATION IN MICE Almaweri AA, Zaitone SA, Al Refai ANM, Hassan MM Email: maweriabdulrazaq@gmail.com Objective: The aim of this study was to assess the toxicity of silver nanoparticle after topical administration on tongue’s mice. Material and Methods: Sixty male and female mice (15.5 − 18.5 g b.w) were randomly divided into the following experimental groups; Group 1 (vehicle control group): consisted of 20 healthy mice (given 0.5 ml distilled water once daily for 28 days topically on mouse’s tongue). Group 2: (low dose group): consisted of 20 healthy mice, (given 0.5 ml nanosilver solution in concentration of 50 g/mlonceg/ml once daily for 28 days topically on mouse’s tongue in a dose of 5mg/kg). Group 3 (high dose group): consisted of 20 healthy mice, (given 0.5 ml nanosilver solution in concentration of 50 g/ mlonceg/ml twice daily for 28 days topically on mouse’s tongue in a dose of 10mg/kg). During 28 days of exposure, the rats were observed weekly for toxicity symptoms. By the end of study, the rats were sacrificed for hematological, biochemical and histopathology studies. The blood samples were drawn then analyzed for ALP (alkaline phosphatase, GOT (glutamic oxaloacetic transaminase), GPT (glutamic pyruvic transminase), CRE (creatinine), and BUN (blood urea nitrogen). Tongue, kidneys, spleen and liver were removed, weighed, fixed, stained, and examined under light microscopy for histopathological study. This study tested the toxicity of silver nanoparticles (20 nm) over a period of 28 days in mice following Organization for Economic Cooperation and Development (OECD) test guideline No. 408 and Good Laboratory Practices (GLP). Results: There was no significant difference in the organs weight of mice after exposure, as well as no significant changes in all four enzymes tested. Histopathology examination revealed no pathological abnormalities in all treated animals compared to untreated healthy animals. (no inflammatory cell infiltration, no hyperplasia, no fatty degeneration ,no edema ,as well as no necrosis). Conclusion: Silver nanoparticles solution in concentration of 50 g/mlonceg/ml is safe when applied topically on tongue’s mice for a period of 28 days. Keywords: Silver; Nanoparticles; Toxicity; Mice. 8 TUIDC 1 Research Article Thamar University International Dental Conference 03 Research Article Research Article Keywords: Silver; Nanoparticles; Toxicity; Rat. TOXICITY EVALUATION OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL APPLICATION IN ALBINO RATS Almaweri AA, Zaitone SA, Al Refai ANM, Hassan MM Email: maweriabdulrazaq@gmail.com Objective: The objective of this study was to evaluate the toxicity of silver nanoparticles after topical administration on tongue of rats. Material and Methods: Sixty males albino rats (180– 200 g b.w.) were randomly divided into the following experimental groups; Group 1 (vehicle control group): consisted of 20 healthy rats (given 0.5 ml distilled water once daily for 28 days topically on ratʼs tongue). Group 2: (nano-silver 50 group): consisted of 20 healthy rats (given 0.5 ml nanosilver solution 50 g/mlonceg/ml once daily for 28 days topically on ratʼs tongue in a dose of 10mg/kg). Group 3 (nano-silver 100 group): consisted of 20 healthy rats (given 0.5 ml nanosilver solution100 g/mlonceg/ml once daily for 28 days topically on ratʼs tongue in a dose of 10mg/kg). During these 28 days of exposure, the rats were observed weekly for toxicity symptoms. By the end of study, the rats were sacrificed for hematological, biochemical and histopathology studies. Blood samples were drawn then analyzed for GOT (glutamic oxaloacetic transaminase), GPT (glutamic pyruvic transminase), CRE (creatinine), and BUN (blood urea nitrogen). Tongue, kidneys, and liver were removed, weighed, fixed, stained, and examined under light microscopy for histopathological test. This study tested the toxicity of silver nanoparticles (20 nm) over a period of 28 days in rats following Organization for Economic Cooperation and Development (OECD) test guideline No. 408 and Good Laboratory Practices (GLP). Results: There was no significant difference in the organs weight of rats after exposure, as well as no significant changes in all four enzymes tested. Histopathology examination revealed no pathological abnormalities in all treated animals compared to untreated healthy animals (no inflammatory cell infiltration, no hyperplasia, no fatty degeneration, no edema, as well as no necrosis). Conclusion: Silver nanoparticle solution in concentrations of 50,100 g/mlonceg/ml is safe when applied topically for a period of 28 days in rats. 9 TUIDC 1 Research Article Thamar University International Dental Conference Research Article Research Article Research Article Keywords: Impaction; Canines; Third molar; Radiographic; Retrospective; Yemen. PREVALENCE AND PATTERN OF IMPACTED TEETH IN YEMENI POPULATION: A RETROSPECTIVE RADIOGRAPHIC STUDY Amran AN, Amran AG, Al-Gabri FA, Al-Basmi AA, Alhajj MN Email: m.n.alhajj@hotmail.com Amran AN, Amran AG, Al-Gabri FA, Al-Basmi AA, Alhajj MN Objective: The aim of this study was to explore the prevalence as well as the pattern of impacted canines, first premolars, and third molars in both maxillary and mandibular jaws in adult Yemeni population. Material and Methods: This was a retrospective radiographic study conducted in Dhamar city, Yemen. For impacted 3rd molars, angular position and depth of impaction were assessed while, for impacted canines the type of impaction was recorded. The angulation was assessed according to winter’s classification and Archer’s classifications. The depth of impaction was recorded according to Pell and Gregory classification. For impacted canines, the type of impaction was assessed according to Al Zoubi’s classification. Data were analyzed and presented in terms of descriptive statistics (frequencies and percentages). Differences between both genders were calculated using Chi-squared test with P-value < 0.05. Results: Records of 913 patients were collected and evaluated for teeth impaction. The most impacted canines were maxillary canines. The most prevalent type of impaction was Type II. Most prevalent impacted 3rd molars were found in the left side of the mandible followed by impacted 3rd molars in the left side of the maxilla. Mesioangular position was the most frequent position of the mandibular impacted 3rd molars followed by vertical position. Conclusion: Impacted mandibular third molars were the most impacted teeth followed by maxillary canines. Prevalence of impacted 3rd molars teeth was more in females than in males while, prevalence of impacted canines was higher in males. 10 TUIDC 1 Research Article Thamar University International Dental Conference 05 Research Article Research Article Keywords: Tooth loss; Yemen; Khat; Shammah; Smoking. ETIOLOGY AND PREVALENCE OF PERMANENT TOOTH LOSS AMONG SAMPLE OF YEMENI POPULATION Alkadasi BA, Al Ahmari NM, Alraawi MA, Noman NA, Aladimi AA, Al-Moaleem MM Email: perio bak@gmail com Objective: To investigate the reasons of the permanent tooth extraction and its relationship with age and gender. Material and Methods: 662 participants, divided into five age groups 14– 23, 24–33, 34– 43, 44–53, and ≤54-years-old. Oral and radiographic examinations were done for each participant. Causes of tooth loss, age group, gender, khat chewing, Shammah use, smoking, teeth brushing and Miswake using were recorded. The data were statically analyzed with SPSS program using Chi-square tests. The p value ≤ 0.050 were considered statistically significant. Results: From the total number of the participants 335(50.6%) were males. The highest age group was in the 14–23 age-group (43.1%). Dental caries was represented by (49.53), while the periodontal disease was (23.3%). The, failure of root canal treatment, orthodontic and other causes were (3.6%, 11.3%, 11.3%), respectively. There were significant differences between genders and different age groups in relation to causes of tooth extraction. Conclusion: Dental caries was the major reason of tooth loss in the young age groups, while the periodontal disease was increased gradually from the middle to the elder age groups among the participants from Taiz, republic of Yemen. Keywords: Tooth loss; Yemen; Khat; Shammah; Smoking. 11 11 TUIDC 1 Research Article Research Article ch Article Thamar University International Dental Conference Thamar University International Dental Conference 06 Research Article Research Article Keywords: Glass-ionomer; ZrO2 nanoparticles; Microhardness; Water sorption. EVALUATION OF SOME MECHANICAL AND PHYSICAL PROPERTIES OF GLASS-IONOMER RESTORATIVE MATERIAL MODIFIED WITH ZIRCONIUM DIOXIDE NANOPARTICLES Alshaibani DAQ, Nassif MS, El-Refai DA Email: Kalidsaeed2013@gmail.com Objective: The objective of this study was to evaluate of some mechanical and physical properties of glass-ionomer restorative material of adding the zirconium dioxide (ZrO2) nanoparticles. Material and Methods: ZrO2nanoparticles were incorporated into the powder component of promedica (Germany) at 3%, 5% and 7% (w/w). Unblended powder was used as control. Mini- flexural strength, compressive strength, were evaluated using a universal testing machine. Surface microhardness was measured using Vickers microhardness tester. Water sorption and solubility was determined as specified in the ISO standard. Data were analyzed using ANOVA and Tukey’s test. Results: GI-containing 3% and 5% (w/w) ZrO2nanoparticles improved the mini-flexural flexural strength and compressive strength compared to the unmodified GI. However, a decrease in the mechanical properties was found for GI-containing 7% (w/w) ZrO2nanoparticles. GI-containing 3% and 5% (w/w) ZrO2nanoparticles improved the surface microhardness. GI-containing 3% and 5% (w/w) ZrO2 nanoparticles improved water sorption and solubility. However, a decrease in the water sorption and solubility was found for GI- containing 7% (w/w) ZrO2 nanoparticles. Conclusions: GI-containing 3% and 5% (w/w) ZrO2 nanoparticles are a promising restorative material with improved mechanical and physical properties. This novel experimental GI may be potentially used for higher stress-bearing site restorations such as Class I and II. 12 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 07 Research Article Research Article Keywords: Chymotrpsin; Oral serratiopetidase; Corticosteroid; Lower third surgery; Postoperative sequelae. EFFICACY OF SUB-MUCOSAL INJECTION OF CHYMOTRYPSIN, ORAL SERRATIOPEPTIDASE AND ORAL CORTICOSTEROIDS FOR REDUCTION POSTOPERATIVE COMPLICATIONS FOLLOWING IMPACTED LOWER THIRD MOLARS SURGERY: A PROSPECTIVE, RANDOMIZED, DOUBLE- BLINDED, CONTROLLED CLINICAL TRIAL Al-Zendani EA, Al-Selwi AM, Al-Moraissi ES Email: dressamalmoraissi@gmail.com Al-Zendani EA, Al-Selwi AM, Al-Moraissi ES Email: dressamalmoraissi@gmail.com Objective: The study aimed to compare between chymotrypsin, oral serratiopeptidase and oral dexamethasone following lower third molars surgery (L3Ms) in respect of postoperative complications. Material and methods: A randomized, double-blinded, clinical trial was conducted on 60 patients who were the candidate to L3Ms and randomly allocated into the following 3 groups: submucosal chymotrypsin, oral serratiopeptidase and oral dexamethasone (each group = 20). The primary outcomes were postoperative pain (visual analogue scale), facial swelling (tape method) and trismus (maximal mouth opening) at 2nd, 5th and 7th postoperative days. Results: All three groups perform a similar achievement following L3MS. There was no statistically significant difference between the three groups in respect of change in facial pain swelling and trismus (P < 0.05). Conclusion: Sub-mucosal injection of chymotrypsin yield comparable effectiveness in decreasing postoperative sequelae following L3MS when compared to oral serratiopeptidase and corticosteroids. This is a first RCT that assessed benefits of sub-mucosal injection of chymotrypsin after L3Ms. Keywords: Chymotrpsin; Oral serratiopetidase; Corticosteroid; Lower third surgery; Postoperative sequelae. 13 13 TUIDC 1 Research Article Thamar University International Dental Conference 08 Research Article Research Article Keywords: Acid etching; Laser etching; Shear bond strength; Orthodontic brackets; Dental fluorosis. EFFECT OF PICOSECOND LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO FLUORESCED TEETH Helmi JJ, Aldhorae K, Altawili Z, Al-Magaleh S, Shamalah A Email: drdurai2008@gmail.com Objective: Active orthodontic treatment with fixed appliances generally last for 2-3 years and successful bonding ensures good treatment progress. Effort are made therefore constantly to improve bond strength , especially in challenging situations such as bonding to gold , porcelain, and amalgam, but the conditions that challenges the orthodontist most is enamel fluorosis. This study aimed to determine if the irradiation of flourized enamel surfaces with picosecond laser might be viable alternative to acid etching to improve the bonding of orthodontic adhesive to flurosed enamel surface. Material and Methods: Sixty four standard edgewise stainless steel brackets (American Orthodontics, Sheboygan, WI, USA) were bonded using Transbond XT (3M Unitek, Germany) on upper flourized premolars prepared by acid etching with 37% phosphoric acid (a), and TimeBandWidth Duetto laser .shear bond strength was measured with a Zwick Universal testing machine at across head speed of 1mm/min. The obtained shear bond strength values of the groups were statically evaluated using SPSS 20 for windows (SPSS Inc. Chicago, IL, USA) using ANOVA, Kolmogorov-Smirnov, and Levine's test at the statistical significance of P <0.05. Results: Mean shear bond strength for acid etching group (13.27MPa), for ultrashort laser group L (18.5 MPa). Conclusion: Picosecond laser etching imparts clinically acceptable shear bond strength of 16.2 MPa. Moreover, laser irradiation play a key role in the caries preventive effects. Keywords: Acid etching; Laser etching; Shear bond strength; Orthodontic brackets; Dental fluorosis. Keywords: Acid etching; Laser etching; Shear bond strength; Orthodontic brackets; Dental fluorosis. 14 14 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 09 Research Article Research Article Keywords: Dental implants; Inferior alveolar nerve; Nerve lateralization. INFERIOR ALVEOLAR NERVE LATERALIZATION FOR DENTAL IMPLANT PLACEMENT IN ATROPHIED POSTERIOR EDENTULOUS MANDIBULAR AREAS Al-Marebi KG Email: khmaribi2012@gmail.com. Objective: The aim of this study was to confirm the effectiveness of inferior alveolar nerve lateralization performed with the piezotome as an alternative treatment for edentulous patients with severe atrophies posterior mandibular areas with evaluation of postoperative neurosensory function and survival rates of implants. Material and Methods: A prospective cohort study was carried out in edentulous patients with posterior mandibular atrophies who were treated between 2016 and 2017. Ten patients were included that have bone height above the IAN was <5 mm, 15 procedures, with 25 implants were placed with an average of 2.5 implants per patient. Panoramic and cone beam computerized tomography radiographic views were taken at 3 months and 1 year after implant loading to check marginal bone loss. The degree of osseointegration of the implants was determined by examining whether or not the implants were both painless and immobile when under torque and while loading. Follow- up visits were done on the third and eighth weeks, and at 6, 12 and 24 months, during each visit through 2 years of study neurosensory function was checked by a two point- discrimination test, in which a measurement of <15 mm was considered normal. Results: The results of this study showed that the normal rate neurosensory function was 95.7% after 8 weeks with implant success rate of 100% were achieved with all 25 implants placed and the implant survival rate was 98.4%. Conclusion: Inferior alveolar nerve lateralization technique performed with piezotome with immediate implant placement can be a useful and safe surgical procedure due to its satisfactory success rate. 15 15 TUIDC 1 Research Article Thamar University International Dental Conference 10 Research Article Research Article Keywords: Acid etching; Laser etching; Bonding; Shear bond strength; Orthodontic brackets. EFFECT OF ULTRASHORT PULSED LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO ENAMEL SURFACE Aldhorae K, Bourauel C, Meister J, Dohrn A, Frentzen M, Joger A, Daratsianos N, Altawilli Z Aldhorae K, Bourauel C, Meister J, Dohrn A, Frentzen M, Joger A, Daratsianos N, Altawilli Z Email: drdurai2008@gmail.com Aldhorae K, Bourauel C, Meister J, Dohrn A, Frentzen M, Joger A, Daratsianos N, Altawilli Z Email: drdurai2008@gmail com Objective: Acid etching is the best method of bonding resins to enamel. A potential disadvantage of enamel acid etching is the demineralization of the most superficial layer, a matter of concern for orthodontist. As a result of demineralization, the surface become more susceptible to long term acid attack and caries. Laser irradiation has been claimed to improve the adhesion of orthodontic adhesive to enamel surface; therefore, it has been proposed as an alternative to acid etching. The study aimed to determine if the irradiation of enamel surfaces with picosecond laser of different output powers might be viable alternative to acid etching for the bonding of orthodontic adhesive to enamel surface. Material and Methods: Sixty upper central incisors standard edgewise stainless steel brackets (American Orthodontics, Sheboygan, WI, USA) were bonded using Transbond XT (3M Unitek, Germany) on bovine incisors prepared by acid etching with 37% phosphoric acid (a), and TimeBandWidth Duetto laser with different settings (L1, L2, L3, L4, L5) .shear bond strength was measured with a Zwick Universal testing machine at across head speed of 1mm/min. The obtained shear bond strength values of the groups were statically evaluated using SPSS 20 for windows(SPSS Inc. Chicago, IL, USA) using ANOVA, Kolmogorov- Smirnov, and Levine's test at the statistical significance of p <0.05. Results: Mean shear bond strength for groups: a (20.1 MPa), L1 (6.7 MPa), L2 (5.9 MPa), L3 (11.1MPa), L4 (15.4MPa), L5 (16.2MPa). Conclusion: Ultrashort pulsed laser etching imparts clinically acceptable shear bond strength (5.9 MPa-16.2 MPa). Keywords: Acid etching; Laser etching; Bonding; Shear bond strength; Orthodontic brackets. 16 16 TUIDC 1 Research Article Thamar University International Dental Conference 11 Research Article Research Article Keywords: Esthetics; Dental students; Laypersons; Perception; Dentofacial; Dentist. PERCEPTION OF ALTERED DENTOFACIAL AESTHETICS BY DIFFERENT DENTAL PROFESSIONALS AND LAYPERSONS Aldhorae K, Alqadasi B, Assiry A, Al-Tawili Z, Shamalah A Email: drdurai2008@gmail.com Objective: Attractive facial and smile features results from the coordination between intra and extraoral soft tissues. This study was aimed to comprehensively assess the perception of altered dentofacial esthetics between different dental professionals and lay people, and to identify the threshold were different variables being to impair dentofacial attractiveness. Material and Methods: Ten photographs were digitally manipulated involving three facial, two smile, four dental, and one gingival component. Fifty images were randomized and rated according to attractiveness by three groups: general dentist, dental students, and lay people. The participants evaluated the original and manipulated images using a visual analogue scale. The responses were then analyzed using Mann-Whitney test. Results: The results demonstrated threshold levels of noticeable differences between varying levels of discrepancy. The overall perception of esthetic was high among dental students with the highest perception toward facial profile and the lowest toward gingival margin height. No differences were found in perception between males and females. Dental students perceived esthetic components more accurately than dentist. Conclusions: Dental students unexpectedly had a better perception of dentofacial esthetics. Unlike gender, clinical training has a substantial positive effect on the assessment of beauty. 17 TUIDC 1 Research Article Thamar University International Dental Conference 12 Research Article Research Article Keywords: Tooth Abnormalities, Orthodontic patients; Radiography, Panoramic. PREVALENCE AND DISTRIBUTION OF DENTAL ANOMALIES AMONG SAMPLE OF ORTHODONTIC AND NON- ORTHODONTIC PATENTS: A RETROSPECTIVE STUDY Aldhorae KA, Altawili ZM, Assiry A, Alqadasi B, Al-Jawfi KA, Hwaiti H Email: drdurai2008@gmail.com Aldhorae KA, Altawili ZM, Assiry A, Alqadasi B, Al-Jawfi KA, Hwaiti H Objective: The study aimed to address the prevalence and distribution of dental anomalies in a group of Yemeni dental patients, and compare the presence of dental anomalies in patients seeking dental and orthodontic treatment. Material and Methods: This retrospective study was conducted on 1675 digital panoramic radiographs for dental patients aged between 9 and 52 years, who visited orthodontic specialty centers in Sana'a city, Yemen, from January 2018 to February 2019. The digital panoramic radiographs were evaluated for prevalence and distribution of dental anomalies bossed on the description presented by White and Pharaoh (Pharaoh and White, 2008). Results: The distribution of anomalies was 30.61% among the orthodontic patients and 22.96% through non – orthodontic patients. The most frequent anomaly among the subjects was impaction (14-47%), macrodontia (11.8%), microdontia (9.23%), hypodontia (7.48%), dilaceration (5,07%), dens evaginatus (1,91%), dens invaginatus (1.58%), hyperdontia (0.99%) and taurodontism (0.91%) respectively. Among the individuals who had dental anomalies 57.9% of subjects showed one type of anomaly, 30.5% had two types of anomalies, while 11,49% had more than two types of anomalies. The selected dental anomalies in present study showed high prevalence rate among the orthodontic patients. Conclusion: Variations in data and results among different studies suggest the impact of racial, genetic and environmental factors. The high frequency of dental anomalies emphasizes the need of early detection, diagnosis which can be achieved through radiographic imaging, this would avoid potential orthodontic, functional and esthetic problems and further emphasizes awareness to minimize any means of complexity of orthodontic and other dental problems. 18 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 13 13 Research Article Research Article Research Article Keywords: Occlusal discrepancies; Interceptive orthodontic; Orthodontic treatment need; Yemen. OCCLUSAL FEATURES AND ORTHODONTIC TREATMENT NEED AMONG HIGH SCHOOL STUDENTS IN SANA'A CITY: A CROSS-SECTIONAL STUDY Al-Hadad, Aldhorae, Al-Tawili Z Email: drdurai2008@gmail.com Objective Dental malocclusion was defined by the World Health Organization (WHO) as the third largest oral health problem that is related to an individual’s general well-being. This cross study was carried out to assess the occlusal features and orthodontic treatment need in Yemeni high school students who had not any orthodontic treatment. Material and methods: A sample of 1036 subjects (527) males and (509) females attending different high schools in scattered areas in Sana’a city. One examiner visited those high schools assisted students using conventional FDI/WHO method of occlusal traits to evaluate occlusal discrepancies and Index of the Orthodontic Treatment Need (IOTN) in order to estimate the need of orthodontic treatment. Collected data was analyzed using the Chi-square test to compare proportions among groups and the significance threshold was set at P < 0.05. Results: Occlusal discrepancies were including: crowding (42%), of whom 16.6% had crowding of ≥ 2mm. spacing was observed in 20.5% of students, mostly in the maxilla. Midline diastema was also found to occur in 9% of students examined. Asymmetrical molar relationship was only observed in 16.1% of students mostly class I/ class II relationship (8%). Overjet was observed in 90.9% of the subjects, 12.8% had deep bite while anterior open bite observed in 3.5%, 12.2% had anterior cross bite whereas 6.6% had posterior cross bite, 1.3% posterior open bite whilst 0.6 had scissor bite. Asymmetrical canine relationship was only observed in 23.8% of students, mostly of class I/ class II relationship (18.3%). Normative orthodontic needs as assessed by criteria of (IOTN) indicated slightly less than half (38.9%) of the students needed some form of orthodontic treatment (Grade 3, 4 and 5). Of this a quarter (24.3%) definitely needed treatment assessment. Conclusion: The findings suggest a need for a more conservative treatment approach in dealing with malocclusion problems among school students in all direction of Yemen. Keywords: Occlusal discrepancies; Interceptive orthodontic; Orthodontic treatment need; Yemen. 19 TUIDC 1 Research Article esearch Article TUIDC 1 Thamar University International Dental Conference Thamar University International Dental Conference 14 Research Article Research Article Keywords: Nasal; Leishmaniasis; Plaque form; Lupoid; Yemen. NASAL LEISHMANIASIS: CLINICO-EPIDEMIOLOGICAL STUDY Alkulaibi MM Email: kulaibim@gmail.com Objective: Cutaneous (CL) and mucocutaneous leishmaniasis (MCL) are the common forms of leishmaniasis affecting the orofacial region. The nasal leishmaniasis is the commonest facial units to be involved in the face. It displays high variety of clinical forms making the disease more difficult to be diagnosed. In Yemen, CL is prevalent However, detailed describing of the nasal lesions is scarce in the literature. Study Area and Population: Seventy six diagnosed cases with nasal leishmaniasis were screened during a CL surveillance in western highlands, Yemen (2015). Demographical data, history, examination with regard to site, size, number, morphology and any associated satellite papules and/or subcutaneous nodule were recorded. Clinical Observation: Patients aged of ≤ 15 years and > 45 years were more frequent accounting for 51%, 33% of the cases respectively. The plaque form was the most prevalent form of the nasal lesion developed commonly (43%) on the nasal lobule of the middle aged and elderly patient. The plaque form was presented in lupoid form, psoriasiform, erysipeloid form and rhinophyma accounting for 45%, 24%, 18%, and 6% of lesions, respectively. The impetiginous form (Figure 10) was only found in four children. 7 cases showed nasal cartilage perforation. Satellite papule and subcutaneous nodules were found in 25 and 4 of the cases, respectively. Conclusion: The plaque form is the most frequent form of nasal leishmaniasis mainly the lupoid form which was found in the nasal lobule in middle-aged and elderly patient. 20 TUIDC 1 Research Article Thamar University International Dental Conference 15 Research Article Keywords: Dental anomalies; Malocclusion; Prevalence; Primary dentition. PREVALENCE OF MALOCCLUSION AND TOOTH ANOMALIES AMONG 3-5-YEAR-OLD PRE- SCHOOL CHILDREN IN SANA’A CITY, YEMEN Yehia LA, Al-Haddad KA, Al-Labani M, Madfa AA Email: dr.lamia.bin.yehia@gmail.com Objective: Malocclusion and dental anomalies are developmental disorders that occur due to the disruption of the process of craniofacial complex growth and odontogenesis respectively. The aim of this study was to investigate the prevalence of malocclusion, dental morphological and numerical anomalies in the primary dentition of randomly chosen three to five-year-old pre-school children in Sana’a City, Yemen. Material and Methods: This was a cross-sectional study. A sample size of 1106 children (boys: 53.3% and girls: 46.7%) were selected. Criteria used by Foster & Hamilton, Björk et al. and Kreiborg et al. with modification done by Carvalho et al. was used to define occlusal traits, malocclusion and dental anomalies respectively. Results: Bilateral flush terminal molar relationship was found in 60%, followed by: bilateral mesial step (27.9%), asymmetrical relationship (8.5%) and distal step (3.5%). Bilateral class I incisor relationship was present in 42.1% followed by: class II (29.3%), class III (23.8%) and open-bite (4.8%). Bilateral canine class I was observed in (62.8%) followed by: asymmetrical relationship (13.6%), class II (12.5%) and class III (11.1%). Normal overjet of 1-3 mm (39.2%), decreased overjet < 1 mm (32.7%), increased over-jet of > 3 mm (16.9%), edge-to edge (8.8%) and reverse bite (2.4%). Ideal over-bite of 1-50% (64.5%) and increased over-bite of > 50% (19.7%). Physiological spacing present in 77.7%. Anthropoid spacing was found in 81.1%. Posterior cross-bite was detected in 29.9%. Dental anomalies: Localized microdontia (15.7%), double teeth (3.3%), hypodontia (0.9%) and hyperdontia (0.7%). Conclusion: Malocclusion was detected in 52.0% of the children. Dental anomalies were detected in 20.7%. Keywords: Dental anomalies; Malocclusion; Prevalence; Primary dentition. 21 TUIDC 1 Research Article h Article TUIDC 1 Thamar University International Dental Conference Thamar University International Dental Conference 16 Research Article Research Article Keywords: Saliva; Human enamel; Pellicle; Erosion; Step height; Proteins; Ions; SDS-PAGE. PROTEINS FROM WHOLE MOUTH SALIVA MEDIATE GREATER PROTECTION AGAINST SEVERE EROSIVE TOOTH WEAR THAN PROTEINS FROM PAROTID SALIVA USING AN IN VITRO MODEL Mutahar M Email: drmahdi2227@yahoo.com Objective: to investigate how the composition of the acquired enamel pellicle (AEP) affected a laboratory model of Erosive tooth wear (ETW) on human enamel by comparing whole mouth saliva (WMS) to parotid saliva (PS). Material and Methods: A total of 60 enamel specimens were prepared from extracted human teeth and were randomly assigned to 4 experimental groups: WMS (n=20), PS (n=20), artificial saliva (AS, n=10) and deionized water (DW, n=10). AEP samples were eluted from WMS (n=5) and PS (n=5) groups after five cycles (acid, wash and saliva) using filter papers soaked in sodium dodecyl sulfate by mechanically rubbing before mean step height was evaluated using a non-contacting profilometer for all groups (n=10 each). Total protein in AEP was quantified using BCA assay, individual protein components of AEP were separated and analyzed using SDSPAGE and western blot for [mucin5b, albumin, carbonic anhydrase VI (CA VI), statherin]. Specific antibody binding was quantified using purified protein standards of known concentration. Samples of AEP were also analyzed by LC/ MS/MS sequencing. Results: WMS group had significantly less acid-induced erosion (step height [4.16 (0.9) μg/ml oncem]) than PS group [6.41 (0.3) μg/ml oncem]. The amount of total protein, mucin5b and albumin were more dominant in WMS pellicles than PS whereas CA VI and statherin were dominant in PS pellicles. Conclusion: The composition of the acquired enamel pellicle influences the degree of protection from acid attack. AEP from WMS was more protective than that of PS, and its likely mechanisms of protection seem to be as a physical barrier rather than stabilizing the crystal structure. 22 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 17 Research Article Research Article Research Article Keywords: Mandibular fracture; Close reduction; Open reduction; Screw; Intra-maxillary fixation. AN EPIDEMIOLOGICAL STUDY ON THE PATTERN AND PREVALENCE OF MANDIBULAR FRACTURE IN HADRAMOUT GOVERNMENT Baraja MM, Al-Moraissi E Email: drmbaraja655@gmail.com Baraja MM, Al-Moraissi E Email: drmbaraja655@gmail.com Objective: The aim of this study was to determine the etiology, frequency of mandibular fractures among different age and sex, to determine the frequency of types of trauma, and to report the different modalities of treatment provided to among patients of AL-WADI Hospital (2004-2017). Material and Methods: A retrospective analysis of the registration files of various mandibular fracture patients was conducted on cases diagnosed during (2004-2017) in patients of AL-WADI Hospital in Hadramout. Results: Out of 43 patients, forty were males (93%) and three were females (7.0%). We found a peak occurrence in young adults, aged 21-40 years (35%). In case of etiology of fracture, road traffic accidents (RTAs) was the most common (58%). In most (60.5%) of the patients, an open reduction and rigid internal fixation using bone plate and screws with intra-maxillary fixation was done. Conclusion: In the present study, the prevalence of mandible fractures was more prevalent in male patients, especially during the 2ed and 3rd decade of life. The most common cause was road traffic accident. Open reduction and rigid internal fixation using miniplates and screws with intra-maxillary fixation was the most commonly used treatment. Keywords: Mandibular fracture; Close reduction; Open reduction; Screw; Intra-maxillary fixation. 23 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 18 Research Article Research Article Keywords: Root filling quality; Dental students; Dental education; Endodontic treatment. PREVALENCE AND ETIOLOGY OF TRAUMATIC DENTAL INJURIES TO ANTERIOR TEETH AMONG PRIMARY SCHOOL CHILDREN IN SANA'A CITY, YEMEN Mutahar NJ, Al-Haddad K Email: dr.nohamutahar@gmail.com Objective: Dental traumatic injuries are common health problems in child and adolescent populations. Information on prevalence and severity of dental trauma in various age groups of the population has significant implications for preventive public oral health care and other strategies for the population. The aims of this study were: a) To determine the prevalence of traumatic dental injuries to anterior teeth among mixed dentition of schoolchildren in Sana'a city, Yemen, and b) To determine the causes of traumatic dental injuries to anterior teeth among mixed dentition of school children in Sana'a City, Yemen. Material and Methods: A cross sectional study was carried out among 1252 school children among (7-12) years old of three districts in Sana’a City using multistage random sampling method. Examination of permanent incisor teeth was done in accordance with the modified Elli’s Classification (By Mc Donald, Avery and Lynch 1983) using a standard mouth mirror and probe. Subjects who had clinical evidence of trauma were interviewed for details of the injury event by using structured questionnaire. Chi square test was used to analyze the distribution of the measurement in this study at the statistical significance of < 0.05. Results: Among the 1252 school children of 30 public and private schools examined, the prevalence of the TDI from total sample size was 129 case (10.30%) with mean age was(10.76 ± 1.22) A statistically significant difference between two groups was found: 10-12 were 4.2 times (95% CI=2.7–6.5) more prone to traumatic dental injuries compared to 7-9 (p<0.001). Experienced TDI, 107 (82.95%) of these were boys and 22 (17.05%) girls. A statistically significant difference between genders was found: boys were 3.83 times (95% CI=2.45–5.98) more prone to traumatic dental injuries compared to girls (p<0.001).The most commonly affected teeth were maxillary central incisors. Public schools were 1.88 times (95% CI=1.3– 2.7) more prone to traumatic dental injuries compared to Private schools (p<0.01). TDI in public schools were (71.32%) which was approximately twice as higher in private schools being (28.68%). Fall was the most common cause for TDI with (62.01%) and most place of occurrence was street (41.08%). Most common type of fracture was class I (51.61%) and most of them were untreated. Conclusion: The level of dental trauma was high but low percentage of children with trauma seeking treatment stresses. AN AUDIT OF ROOT CANAL FILLING QUALITY PERFORMED BY UNDERGRADUATE PRE- CLINICAL DENTAL STUDENTS, YEMEN (ACCEPTED IN BMC MEDICAL EDUCATION, ARTICLE NUMBER: 350 (2019)) Al-Anesi MS, Alkhawalani MM, Alkheraif AA, Al-Basmi AA, Alhajj MN Email: m.n.alhajj@hotmail.com Objective: Dental students are future dentists. Continuous assessment and improving of the educational curricula will ensure excellent academic performance of dental students and thus providing the community with the best treatment modalities. The aim of this study was to evaluate the root canal filling quality performed in extracted teeth by preclinical undergraduate Yemeni dental students. Material and Methods: Root canal treatment was performed by undergraduate preclinical dental students on 331 extracted human teeth including 741 roots. The teeth were then collected and evaluated radiographically based on three criteria of quality (length, density, and taper). Cohen’s Kappa test was used to assess the agreement between the examiners and Chi-squared test was used for the association between the study variables. The level of significant was set at α <0.05. Results: The results of the study revealed that the overall quality of roots canals fillings was poor. However, more than half of the study sample (53.4%) had adequate length, 13.1% had adequate density, and 14.2% had adequate taper. Anterior as well as single-rooted teeth had significantly better quality than posterior and multi-rooted teeth, respectively. The root canal fillings quality of mandibular teeth was better than that of maxillary teeth with no significant difference (P> 0.05). Conclusion: The findings of the study emphasize the need of improving the endodontic course in the preclinical level and more advanced techniques and instruments should be incorporated. Keywords: Root filling quality; Dental students; Dental education; Endodontic treatmen 24 TUIDC 1 Research Article Thamar University International Dental Conference 19 Research Article Research Article Keywords: Traumatic dental injuries; Incisors; Fractures. PREVALENCE AND ETIOLOGY OF TRAUMATIC DENTAL INJURIES TO ANTERIOR TEETH AMONG PRIMARY SCHOOL CHILDREN IN SANA'A CITY, YEMEN We need for increased awareness in Sana’a population to prevent future complications and high cost treatments. It is importance that all traumatic injuries are diagnosed, treated, and controlled in time 25 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 20 Research Article Keywords: Corrosion resistance; Ni-Cr alloys; Mouthwash; Artificial saliva. INFLUENCE OF CHLORHEXIDINE MOUTHWASHES ON CORROSION RESISTANCE OF NI–CR DENTAL CASTING ALLOYS Alsubari RA, Merzouk N Email: ras800_as@yahoo.com Objective: Metals and alloys have many uses in dentistry. The most commonly used base metal alloys in dentistry are nickel–chromium alloy, which are commonly used for crown and bridge casting. The most important factors that affect the choice of dental metallic alloys are the body acceptability for these alloys, or the so-called biocompatibility. The aim of the present study was to compare the corrosion behavior of the Ni-Cr alloys used for crown and bridge casting in three mouthwashes, in relation to a reference solution, namely Fusayama Meyer saliva, in order to assess the influence of these products on the corrosion resistance of the Ni– Cr dental alloys. Material and methods: The three materials analyzed were Ni-Cr dental alloys: Kera NC®, Realloy N® and 4 all®, which were tested in three mouthwashes: Hexa®, Hexadyle® and Coxydil® as well as in Fusayama-Meyer artificial saliva. The electrochemical measurements were carried out at room temperature, a potentiostat (PGZ 100 France radiometer analytical S.A) was used to perform the electrochemical measurements. Results: The corrosion behavior study showed that in Hexa® mouthwash the 4 all® alloy was the least resistance to corrosion, in Fusayama-Meyer artificial saliva tended to be more corrosive for Kera NC® alloy, in Coxydil® mouthwash exhibited the highest corrosion resistance for Kera NC®, and 4 all® alloys, in Coxydil® and Hexadyle® mouthwashes exhibited the highest corrosion resistance for Realloy N® alloy. Conclusion: The results obtained in the present study will enable us to provide attending practitioners with advice concerning mouthwash to recommended, depending on the treatment phase and the alloy used. So we can advise Coxydil® mouthwash to patients treated with 4 all® and Kera NC® alloys, Coxydil® or Hexadyle® mouthwashes for the patients treated with Realloy N® alloy. 26 TUIDC 1 Research Article Thamar University International Dental Conference 21 Research Article Research Article Keywords: Periodontal disease; Khat chewing; Smoking; Yemeni; Risk factors. ASSOCIATION OF KHAT CHEWING AND OTHER RISK FACTORS WITH PERIODONTAL DISEASES AMONG ADULT DENTAL PATIENTS Alhajj WA, Hwaiti H, Shamala A, Al-Azazi H, Alwesabi M Email: waddah.alhaj@gmail.com Objective: Periodontal diseases are very common dental disease. Many risk factors may play significant role in the periodontal disease initiation and progression. This study was performed to evaluate the effects of khat chewing, smoking, age and sex as risk factors on clinical parameters of periodontal diseases among Yemeni dental patients. Material and Methods: This cross-sectional study was performed on 1231 patients attending the outpatient dental polyclinics of University of Science and Technology during the academic years 2017/2018. Preformed case sheet was used as a tool of data collection in the clinical sessions. All data concerning this study were revised carefully by the supervisors during data collection. All completed sheets were collected throughout the year by the supervisors, then the data were managed using SPSS statistical package. Data cleaning, descriptive statistics, and inferential statistics were then performed. Results: Khat chewers were higher among the study sample than non-chewers counterparts (60.7% vs. 39.3%). Regarding smoking, prevalence of smoking among the study sample was 25.5% (297 patients). Study results indicated that periodontitis are more associated with khat chewing, smoking, female gender and age older than 35 years old. Results also showed that female and age > 35 years were significantly associated with gingival recession. Mean number of the teeth with gingival recession in male patients were more teeth than in females. Regarding furcation involvement, khat chewing, smoking, male gender, and patients > 35 years old were significantly associated with furcation involvement. Conclusion: It was concluded from the present study that khat chewing, smoking, age older than 35 seem to be risk factors of periodontal diseases. 27 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 22 Research Article Research Article Keywords: Khat; Melanin; Risk; Smoking; Yemen. PREVALENCE OF MELANIN PIGMENTATION IN A YEMENI POPULATION AND ITS RELATION TO SOME RISK FACTORS Alhajj WA, Alhajj MN Email: waddah.alhaj@gmail.com Objective: The present study aimed to explore the prevalence of melanin pigmentation in a sample of Yemeni population and its relation to some possible risk factors. Material and Methods: In this cross-sectional study printed questionnaires were introduced to the patients and a clinical examination was performed for each patient. The questionnaire included questions regarding demographic data, and questions regarding some common habits such as smoking, khat chewing and consuming of hot drinks. Melanin pigmentation was assessed regarding its presence, most affected areas and the numbers of affected quadrants. The data were managed and analyzed using SPSS software program at P value < 0.05. Results: Four hundred and forty patients with mean age 29 ± 8.21 years were included in the study. Of them, 67.5% were fair-skinned, 26.8% were smokers, 47.5%” were khat chewers and 33.6% were hot drinks consumers. The prevalence of melanin pigmentation was 62.7%, with class I represented 56.9% of cases. Males showed more prevalence (67.9%) of melanin pigmentation than females (57.7%) with no significant difference. Results also showed more prevalence of melanin pigmentation in patients > 25 years, and dark-skinned patients. Regarding habits, smoking, khat chewing and hot drink consumption habits showed significant associations with melanin pigmentation. Whereas the association of khat chewing and hot drinks alone with melanin pigmentation showed no significant relationship. Conclusion: It can be concluded that Yemeni people had high prevalence of melanin pigmentation with more prevalence of CL I type. Males, patients > 25 years and dark-skinned patients showed more prevalence of melanin pigmentation. Smoking, khat chewing and hot drinks consuming habits had significant associations with melanin pigmentation. 28 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 23 23 Research Article Research Article Research Article Keywords: Apical abscess; Apical periodontitis; Knowledge; Dental practitioners. ATTITUDE AND KNOWLEDGE OF A YEMENI DENTAL PRACTITIONERS TOWARDS MANAGEMENT OF ACUTE APICAL PERIODONTITIS Altawili ZM, Madfa AA Email: dentzee7@gmail.com Objective: Management of acute apical periodontitis is almost every day dental practice, planning for proper outcome for endodontic therapy depends on various factors to maintain the quality of root canal treatment standards including both restoration of root canal system and coronal part, and individual factors including dentist knowledge, attitude and skills. Therefore, the purpose of this study was to address the current knowledge and attitude of Yemeni dental practitioner employed during root canal therapy of tooth associated with acute apical periodontitis. Material and Methods: This cross-sectional study was conducted randomly on 146 general practitioners, who practiced in dental clinics in Sana'a city, Yemen. The collected data was analyzed using the Chi square test and the significance threshold was set at P < 0.05. Results: Of the respondents 93.58% were prescribed antibiotic and 78.2% were anesthetized associated tooth. The low usage of rubber dam was strictly prevalence only 0.64% of the participants were used rubber dam compared to other isolation methods. Among the participants, 57.05% used both hand and rotary means in cleaning and shaping the root canal system. Multiple irrigation protocol was the most followed protocol (69.23%), of whom 39.1% were used Sodium hypochlorite, hydrogen peroxide and chlorhexidine successively. Of the respondents, the mixture of both calcium hydroxide and chlorhexidine (57.05%) as intracanal medicament was more frequent than Calcium Hydroxide (48.07%). Conclusion: the result of this study indicates that the recently graduated dental practitioners were following the recommended standard of endodontic treatment better than those of longer time from the graduation. Keywords: Apical abscess; Apical periodontitis; Knowledge; Dental practitioners. 29 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 24 Research Article Research Article Keywords: Mandibular angle fractures; Mandibular condyle fractures; Mandibular third molars; Occlusal support. THE EFFECT OF THE LOWER WISDOM TOOTH AND OCCLUSAL SUPPORT ON MANDIBULAR ANGLE AND CONDYLE FRACTURES Bin Z, Al-Sharani H, Falci SGM, Jing M Email: hishamm2010@live.com Objective: This retrospective study was intended to assess whether the mandibular angular and condylar fractures are influenced by the existence or absence of the lower third molar (angulation & impaction) as well as the occlusal support. Material and Methods: The study was performed among patients who experienced mandibular fractures in the department of maxillofacial surgery at the three affiliated hospitals of Jinzhou Medical University, Jinzhou, China. The study covered 73 fractured jaws (44 angular & 29 condylar fractures). Data were selected randomly from 2015 till 2019. Patient's medical charts and radiographs were the exact source for data acquisition. The independent variables were the existence or absence of lower wisdom tooth and occlusal support. The system of Pell and Gregory has been applied to assess the various positions of lower impacted M3 while Winter’s classification was used to evaluate its angulations. The dependent variables were the resultant fractures whether angle or condyle. Data analysis was carried out by applying Fisher’s exact test and χ2 distribution test with (p< 0.05) level of significance. Results: The presence of lower M3 or occlusal support is significantly associated with the raised prevalence of angular fractures and conversely diminished the risk of fractures in the condyle. Conclusion: The lack of lower M3 or occlusal support decreases the frequency of angular fractures occurrence and conversely increase the incidence of fractures in the condyle. Keywords: Mandibular angle fractures; Mandibular condyle fractures; Mandibular third molars; Occlusal support. 30 TUIDC 1 Research Article Thamar University International Dental Conference 25 Research Article Research Article Keywords: Full hard stabilization splint; Anterior repositioning splint; Full soft stabilization splint; Arthrogneous TMDs; Myogenus TMDs. DO OCCLUSAL SPLINTS THERAPY CURE TEMPOROMANDIBULAR DISORDERS? Al-Moraissi E Email: dr_essamalmoraissi@yahoo.com Objective: A network meta-analysis (NMA) of randomized clinical trials (RCTs) was performed to assess the effectiveness of various types of occlusal splints and rank them according to their effectiveness in managing TMDs. Material and Methods: An electronic search was undertaken to identify RCTs published until Augst 2019. Predictor variables were control, non-occluding splints, hard stabilisation splint (SS), soft SS, prefabricated splint, minianterior splints, anterior repositioning splint (ARS), and counseling therapy (CT) with and without hard SS. Outcome variables were pain improvement, posttreatment pain intensity, improvement in mouth opening and TMJ sounds. Results: Forty-nine RCTs involving 2000 patients were included. There was a significant decrease in posttreatment pain intensity in arthrogenous TMDs after ARS (low-quality evidence), CT plus hard SS (moderate-quality evidence), minianterior splints (very low-quality evidence)) and hard SS alone (low-quality evidence) when compared to control. There was s significant decrease in posttreatment pain intensity of myogenous TMDs with minianterior splints (very low-quality evidence), soft SS (very low-quality evidence), CT alone (moderate- quality evidence), CT plus hard SS (moderate-quality evidence), and hard SS alone (moderate- quality evidence)., when compared to control. ARS and CT were superior in decreasing TMJ clicking than control and hard SS alone. Conclusion: Based on the current NMA of 49 RCTs, there is a moderate to very low-quality evidence confirming the effectiveness of occlusal splints therapy in the treatment of TMDs. Multimodal therapy consisting of CT & self- care plus hard SS could produce the maximum improvement for TMDs patients. Keywords: Full hard stabilization splint; Anterior repositioning splint; Full soft stabilization splint; Arthrogneous TMDs; Myogenus TMDs. 31 TUIDC 1 Research Article Thamar University International Dental Conference 26 Research Article Research Article NEEDLING THERAPIES IN THE MANAGEMENT OF TEMPOROMANDIBULAR MYOFASCIAL PAIN: A NETWORK META-ANALYSIS OF RANDOMIZED CLINICAL TRIALS Al-Moraissi E, Aladashi O, Goddard G, Christidis N Email: dr_essamalmoraissi@yahoo.com Objective: A network meta-analysis (NMA) of randomized clinical trials (RCTs) was performed aiming to compare the treatment outcome of dry needling, acupuncture, or wet needling using different substances in managing myofascial pain in the orofacial region (TMD- M). Material and Methods: An electronic search was undertaken to identify randomized clinical trials (RCTs) published until September 2019, comparing dry needling, acupuncture, or wet needling using local anaesthesia (LA), botulinum toxin-A (BTX-A), granisetron, platelet-rich plasma (PRP) or passive placebo versus real active placebo in patients with TMD-M. RCTs meeting the inclusion criteria were stratified according to the follow-up time; immediate post- treatment to 3 weeks, and one to 6 months post-treatment. Outcome variables were post- treatment pain intensity, increased mouth opening (MMO) and pressure threshold pain (PPT). Mean difference (MD) was used to analyzed via frequentist NMA using STATA software. Results: Twenty-two RCTs involving 959 patients were included. There was significant pain decrease after PRP when compared to an active/passive placebo and acupuncture. There was a significant improvement of MMO after LA (MD=3.65; CI: 1.18-6.1) and dry needling therapy (MD= 2.37; CI: 0.66-4) versus placebo. Conclusion: Based on this NMA, one can conclude that the effectiveness of needling therapy did not depend on needling type (dry or wet) or needling substance. Also, the present NMA supported the hypothesis that needling therapies have an antinociceptive effect. An effect of needling therapy was not beyond the placebo effect. Keywords: Wet needling therapy; Dry needling; Acupuncture; Temporomandibular disorders; Myofascial pain. 32 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 27 Research Article Research Article Keywords: Conservative treatments; Arthrocentesis; Platelet-rich plasma; Hyaluronic acid; Arthroscopy; Physical therapy; TMJ pain. WHAT IS THE MOST EFFECTIVE TREATMENT FOR ARTHROGENOUS TEMPOROMANDIBULAR DISORDERS? Al-Moraissi E Email: dr_essamalmoraissi@yahoo.com Objective: We conducted a NMA of randomized clinical trials (RCTs) to identify the most effective treatment of arthrogenous TMDs with respect to pain reduction and improved mouth opening. Material and Methods: An electronic search on three major databases was undertaken to identify RCTs published before August 2019, comparing up to fourteen different treatments against control/placebo patients for arthrogenous TMDs with respect to pain reduction and improved mouth opening. The treatment variables were controls/placebo, conservative treatment (muscle exercises and occlusal splint therapy), occlusal splint therapy alone, intraarticular injection (IAI) of hyaluronic acid (HA) or corticosteroid (CS), arthrocentesis with or without HA, CS and platelet-rich plasma, (PRP), arthroscopy with or without HA and PRP, open joint surgery, and physiotherapy. Results: Thirty-six RCTs were identified that performed comparative outcome assessments for pain and 33 RCTs for MMO. At the short term (≤ 5 months), IAI-HA and IAI-CS achieved a substantially greater pain reduction than control/placebo. At intermediate term (≥ 6 months), a statistically significant decrease in posttreatment pain intensity was observed following Arthroscopy-PRP, Arthrocentesis-PRP, Arthroscopy-, TMJ- surgery, IAI-HA, Arthroscopy- alone and Arthrocentesis-HA when compared to the control /placebo groups. Conclusion: There is a very low to moderate quality level evidence supporting that minimally invasive procedures, particularly in combination with IAI of adjuvant pharmacological agents (PRP, HA or CS), are significantly more effective than conservative treatments for both pain reduction and improvement of MMO in both short (≤ 5 months) and intermediate term (6 months to 4 years) periods. Keywords: Conservative treatments; Arthrocentesis; Platelet-rich plasma; Hyaluronic acid; Arthroscopy; Physical therapy; TMJ pain. 33 TUIDC 1 Research Article Thamar University International Dental Conference 28 28 Research Article Research Article Keywords: Endosequence root repair material; iRoot BP; iRoot BP Plus; Pulp capping. EVALUATION OF THE SAME COMPOSITION MATERIALS: IROOT BP PLUS AND ENDOSEQUENCE ROOT REPAIR MATERIAL (ERRM) AS PULP CAPPING AGENTS: A SYSTEMIC REVIEW Mahgoub NM, Alqadasi B, Aldhorae K, Assiry A, Altawili ZM, Hong T Email: drdurai2008@gmail.com Objective: iRoot BP Plus or Endosequence Root Repair Material (EERM) is a premixed bioceramic thick/putty. According to its instruction manual, iRoot BP Plus is composed of Tricalcium silicate, Zirconium oxide, Tantalum pentoxide, Dicalcium silicate, Calcium sulfate, Calcium phosphate monobasic and filler agents. This systematic review was aimed to evaluate and present the iRoot BP Plus material as a pulp capping agent. Material and Method: A systematic search for articles with the scope of the selection criteria undergoing for data extraction was conducted through electronic databases. Studies on evaluation of the cytotoxicity, bioactivity and dentinal bridge formation of iRoot BP, iRoot BP Plus, Endosequence Root Repair Material putty or paste (ERRM) on variant human cells were selected for In-vitro models, and dentinal bridge formation on human and animals’ teeth for In-Vivo models were selected. Results: A total of 22 articles were discussed in the review; 14 in-vitro studies, five in-vivo studies, and three articles with both studies. MTT was the most used method for evaluating cytotoxicity. As for dentinal bridge formation, histological assessment and Micro-CT were used. Human Dental Pulp Cells were the most investigated for In-vitro models and Rats for In- vivo models. Except for one study, all studies involved in this review were primarily examining the material and comparing it to different types of MTA. Conclusion: iRoot BP, iRoot BP Plus, and ERRM are biocompatible materials that enhance Human Dental Pulp Cells' (hDPCs) and other variant human cells proliferation, migration, attachment adhesion, mineralization, and dentinal bridge formation. eywords: Endosequence root repair material; iRoot BP; iRoot BP Plus; Pulp capping. 34 TUIDC 1 Research Article Research Article earch Article TUIDC 1 Thamar University International Dental Conference Thamar University International Dental Conference Research Article Keywords: Amalgam; Composite; Repair; Veneer; Minimal invasive. MINIMAL INVASIVE DENTISTRY: A CLINICAL CASE SERIES REPORT Alsanabani F, Madfa A, Alhamzi M, Alqudaimi G Email: fsanabani@yahoo.com Dental composite restorative materials are newly products in the dental market when compared to the old history of using amalgam restorative materials. However, dental composites are used commonly in the modern dentistry as posterior restorations replacing amalgam because of their esthetic appearance and adhesive properties. Despite of that, dental composite has some drawbacks that limit its using in some cases such as large stress bearing involvement or dentinal margins. On the other hand, dental practitioners should response to the esthetic demands of their patients, at the same time the longevity of restoration and minimal intervention dentistry concept should be considered. Since there is no ideal restorative material existing until now, the aim of this clinical case report series is to present a combination of the advantages of both composite and amalgam materials reflecting minimal invasive dentistry with enhancement of the longevity of the restoration. The clinical techniques using composite to veneer the amalgam restoration or repair rather than replace are easy to apply with low failure risk factors, low cost and low time consuming. 35 TUIDC 1 Research Article Research Article Thamar University International Dental Conference 30 Research Article Research Article Keywords: Cuspid; Dental Prosthesis, Implant-Supported; Dental Implantation, Endosseous. DENTAL IMPLANTS FOR REPLACEMENT BILATERAL CONGENITALLY MISSING MAXILLARY CANINES: THREE YEARS FOLLOW-UP Alsameai AM, Khabaz BA, Al-Moaleem MM Maxillary canines play strategic roles in maintaining the normal function and visibility of teeth. However, rare cases of bilateral permanent congenitally missing maxillary canines were occurring. Replacing these missing canines with dental implant prostheses is an optimal treatment plan for aesthetics and functions. In this case report, we described the replacement of bilateral maxillary canines with osseointegrated implants to support all-ceramic prostheses for a young female patient. This treatment avoids the conventional preparation of adjacent teeth as a part of prosthetic reconstructions. In addition to that all-ceramic crowns are biocompatible with oral cavity structures. Stable short-term result has been achieved with the replacement of the bilateral congenitally missed canines with an implant and cemented all- ceramic crowns, with 3-years cumulative success rate. Keywords: Cuspid; Dental Prosthesis, Implant-Supported; Dental Implantation, Endosseous. 36 INDEXS TUIDC 1 Thamar University International Dental Conference INDEX - Title AN AUDIT OF ROOT CANAL FILLING QUALITY PERFORMED BY UNDERGRADUATE PRE- CLINICAL DENTAL STUDENTS, YEMEN (ACCEPTED IN BMC MEDICAL EDUCATION, ARTICLE NUMBER: 350 (2019)), 18 AN EPIDEMIOLOGICAL STUDY ON THE PATTERN AND PREVALENCE OF MANDIBULAR FRACTURE IN HADRAMOUT GOVERNMENT, 17 ASSOCIATION OF KHAT CHEWING AND OTHER RISK FACTORS WITH PERIODONTAL DISEASES AMONG ADULT DENTAL PATIENTS, 21 ATTITUDE AND KNOWLEDGE OF A YEMENI DENTAL PRACTITIONERS TOWARDS MANAGEMENT OF ACUTE APICAL PERIODONTITIS, 23 DENTAL IMPLANTS FOR REPLACEMENT BILATERAL CONGENITALLY MISSING MAXILLARY CANINES: THREE YEARS FOLLOW-UP, 30 DO OCCLUSAL SPLINTS THERAPY CURE TEMPOROMANDIBULAR DISORDERS?, 25 EFFECT OF PICOSECOND LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO FLUORESCED TEETH, 8 EFFECT OF ULTRASHORT PULSED LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO ENAMEL SURFACE,10 AN AUDIT OF ROOT CANAL FILLING QUALITY PERFORMED BY UNDERGRADUATE PRE- CLINICAL DENTAL STUDENTS, YEMEN (ACCEPTED IN BMC MEDICAL EDUCATION, ARTICLE NUMBER: 350 (2019)), 18 AN EPIDEMIOLOGICAL STUDY ON THE PATTERN AND PREVALENCE OF MANDIBULAR FRACTURE IN HADRAMOUT GOVERNMENT, 17 ASSOCIATION OF KHAT CHEWING AND OTHER RISK FACTORS WITH PERIODONTAL DISEASES AMONG ADULT DENTAL PATIENTS, 21 ATTITUDE AND KNOWLEDGE OF A YEMENI DENTAL PRACTITIONERS TOWARDS MANAGEMENT OF ACUTE APICAL PERIODONTITIS, 23 DENTAL IMPLANTS FOR REPLACEMENT BILATERAL CONGENITALLY MISSING MAXILLARY CANINES: THREE YEARS FOLLOW-UP, 30 DO OCCLUSAL SPLINTS THERAPY CURE TEMPOROMANDIBULAR DISORDERS?, 25 EFFECT OF PICOSECOND LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO FLUORESCED TEETH, 8 EFFECT OF ULTRASHORT PULSED LASER VERSUS CONVENTIONAL ACID ETCHING ON SHEAR BOND STRENGTH OF ORTHODONTIC BRACKETS BONDED TO ENAMEL SURFACE,10 EFFICACY OF SUB-MUCOSAL INJECTION OF CHYMOTRYPSIN, ORAL SERRATIOPEPTIDASE AND ORAL CORTICOSTEROIDS FOR REDUCTION POSTOPERATIVE COMPLICATIONS FOLLOWING IMPACTED LOWER THIRD MOLARS SURGERY: A PROSPECTIVE, RANDOMIZED, DOUBLE-BLINDED, CONTROLLED CLINICAL TRIAL, 7 ESTABLISHMENT OF CEPHALOMETRIC NORMS FOR YEMENI ADULTS AND COMPARING IT WITH CAUCASIAN NORMS, 1 IT WITH CAUCASIAN NORMS, 1 ETIOLOGY AND PREVALENCE OF PERMANENT TOOTH LOSS AMONG SAMPLE OF YEMENI POPULATION,5 YEMENI POPULATION,5 EVALUATION OF SOME MECHANICAL AND PHYSICAL PROPERTIES OF GLASS-IONOMER RESTORATIVE MATERIAL MODIFIED WITH ZIRCONIUM DIOXIDE NANOPARTICLES, 6 EVALUATION OF THE SAME COMPOSITION MATERIALS: IROOT BP PLUS AND ENDOSEQUENCE ROOT REPAIR MATERIAL (ERRM) AS PULP CAPPING AGENTS: A SYSTEMIC REVIEW, 28 NFERIOR ALVEOLAR NERVE LATERALIZATION FOR DENTAL IMPLANT PLACEMENT IN TROPHIED POSTERIOR EDENTULOUS MANDIBULAR AREAS, 9 INFERIOR ALVEOLAR NERVE LATERALIZATION FOR DENTAL IMPLANT PLACEMENT IN ATROPHIED POSTERIOR EDENTULOUS MANDIBULAR AREAS, 9 NFLUENCE OF CHLORHEXIDINE MOUTHWASHES ON CORROSION RESISTANCE OF NI– R DENTAL CASTING ALLOYS, 20 MINIMAL INVASIVE DENTISTRY: A CLINICAL CASE SERIES REPORT, 29 PERCEPTION OF ALTERED DENTOFACIAL AESTHETICS BY DIFFERENT DENTAL PROFESSIONALS AND LAYPERSONS, 11 37 TUIDC 1 Thamar University International Dental Conference INDEX - Title PREVALENCE AND DISTRIBUTION OF DENTAL ANOMALIES AMONG SAMPLE OF ORTHODONTIC AND NON- ORTHODONTIC PATENTS: A RETROSPECTIVE STUDY, 12 PREVALENCE AND ETIOLOGY OF TRAUMATIC DENTAL INJURIES TO ANTERIOR TEETH AMONG PRIMARY SCHOOL CHILDREN IN SANA'A CITY, YEMEN, 19 PREVALENCE AND PATTERN OF IMPACTED TEETH IN YEMENI POPULATION: A RETROSPECTIVE RADIOGRAPHIC STUDY, 4 PREVALENCE OF MALOCCLUSION AND TOOTH ANOMALIES AMONG 3-5-YEAR-OLD PRE- SCHOOL CHILDREN IN SANA’A CITY, YEMEN, 15 PREVALENCE OF MELANIN PIGMENTATION IN A YEMENI POPULATION AND ITS RELATION TO SOME RISK FACTORS, 22 PROTEINS FROM WHOLE MOUTH SALIVA MEDIATE GREATER PROTECTION AGAINST SEVERE EROSIVE TOOTH WEAR THAN PROTEINS FROM PAROTID SALIVA USING AN IN VITRO MODEL, 16 THE EFFECT OF THE LOWER WISDOM TOOTH AND OCCLUSAL SUPPORT ON MANDIBULAR ANGLE AND CONDYLE FRACTURES, 24 TOXICITY ASSESSMENT OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL ADMINISTRATION IN MICE, 2 TOXICITY EVALUATION OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL APPLICATION IN ALBINO RATS, 3 WHAT IS THE MOST EFFECTIVE TREATMENT FOR ARTHROGENOUS TEMPOROMANDIBULAR DISORDERS?, 27 PREVALENCE AND DISTRIBUTION OF DENTAL ANOMALIES AMONG SAMPLE OF ORTHODONTIC AND NON- ORTHODONTIC PATENTS: A RETROSPECTIVE STUDY, 12 PREVALENCE AND ETIOLOGY OF TRAUMATIC DENTAL INJURIES TO ANTERIOR TEETH AMONG PRIMARY SCHOOL CHILDREN IN SANA'A CITY, YEMEN, 19 PREVALENCE AND PATTERN OF IMPACTED TEETH IN YEMENI POPULATION: A RETROSPECTIVE RADIOGRAPHIC STUDY, 4 PREVALENCE OF MALOCCLUSION AND TOOTH ANOMALIES AMONG 3-5-YEAR-OLD PRE- SCHOOL CHILDREN IN SANA’A CITY, YEMEN, 15 PREVALENCE OF MELANIN PIGMENTATION IN A YEMENI POPULATION AND ITS RELATION TO SOME RISK FACTORS, 22 PROTEINS FROM WHOLE MOUTH SALIVA MEDIATE GREATER PROTECTION AGAINST SEVERE EROSIVE TOOTH WEAR THAN PROTEINS FROM PAROTID SALIVA USING AN IN VITRO MODEL, 16 THE EFFECT OF THE LOWER WISDOM TOOTH AND OCCLUSAL SUPPORT ON MANDIBULAR ANGLE AND CONDYLE FRACTURES, 24 TOXICITY ASSESSMENT OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL ADMINISTRATION IN MICE, 2 TOXICITY EVALUATION OF SILVER NANOPARTICLES SOLUTION AFTER TOPICAL APPLICATION IN ALBINO RATS, 3 WHAT IS THE MOST EFFECTIVE TREATMENT FOR ARTHROGENOUS TEMPOROMANDIBULAR DISORDERS?, 27 ADMINISTRATION IN MICE, 2 38 INDEX - Author TUIDC 1 Thamar University International Dental Conference 5 2 3 26 5 21 4 18 Al Ahmari NM Al Refai ANM Al Refai ANM Aladashi O Aladimi AA Al-Azazi H Al-Basmi AA Al-Basmi AA Aldhorae 13 8 28 Aldhorae K Aldhorae K Al-Gabri FA 4 19 Al-Haddad K Al-Haddad KA 15 4 18 Alhajj MN Alhajj MN Alhajj MN 22 29 12 18 18 15 1 8 5 30 17 7 11 12 28 29 5 7 24 8 11 13 12 28 10 21 Alhamzi M Al-Jawfi KA Alkhawalani MM Alkheraif AA Al-Labani M Al-Labani MA Al-Magaleh S Al-Moaleem MM Al-Moaleem MM Al-Moraissi E Al-Moraissi E Alqadasi B Alqadasi B Alqadasi B Alqudaimi G Alraawi MA Al-Selwi AM Al-Sharani H Altawili Z Al-Tawili Z Al-Tawili Z Altawili ZM Altawili ZM Altawilli Z Alwesabi M Amran AG 4 11 12 Assiry A Assiry A Assiry A 28 10 26 10 10 6 Bourauel C Christidis N Daratsianos N Dohrn A El-Refai DA 10 26 2 3 29 12 21 24 10 30 29 15 23 10 20 Frentzen M Goddard G Hassan MM Hassan MM Hong T Hwaiti H Hwaiti H Jing M Joger A Khabaz BA Madfa A Madfa AA Madfa AA Meister J Merzouk N Nassif MS 6 5 21 8 11 Noman NA Shamala A Shamalah A Shamalah A Zaitone SA 2 3 18 10 Zaitone SA Al-Anesi MS Aldhorae K Aldhorae K 11 12 13 21 22 Aldhorae KA Al-Hadad Alhajj WA Alhajj WA Alkadasi BA 5 14 9 2 Alkulaibi MM Al-Marebi KG Almaweri AA Almaweri AA 3 27 26 25 30 29 6 20 23 1 7 4 17 24 8 28 16 19 Al-Moraissi E Al-Moraissi E Al-Moraissi E Alsameai AM Alsanabani F Alshaibani DAQ Alsubari RA Altawili ZM Al-Yousefi AA Al-Zendani EA Amran AN Baraja MM Bin Z Helmi JJ Mahgoub NM Mutahar M Mutahar NJ Thamar University International Dental Conference 5 2 3 26 5 21 4 18 Al Ahmari NM Al Refai ANM Al Refai ANM Aladashi O Aladimi AA Al-Azazi H Al-Basmi AA Al-Basmi AA Aldhorae 13 8 28 Aldhorae K Aldhorae K Al-Gabri FA 4 19 Al-Haddad K Al-Haddad KA 15 4 18 Alhajj MN Alhajj MN Alhajj MN 22 29 12 18 18 15 1 8 5 30 17 7 11 12 28 29 5 7 24 8 11 13 12 28 10 21 Alhamzi M Al-Jawfi KA Alkhawalani MM Alkheraif AA Al-Labani M Al-Labani MA Al-Magaleh S Al-Moaleem MM Al-Moaleem MM Al-Moraissi E Al-Moraissi E Alqadasi B Alqadasi B Alqadasi B Alqudaimi G Alraawi MA Al-Selwi AM Al-Sharani H Altawili Z Al-Tawili Z Al-Tawili Z Altawili ZM Altawili ZM Altawilli Z Alwesabi M Amran AG 4 11 12 Assiry A Assiry A Assiry A 28 10 26 10 10 6 Bourauel C Christidis N Daratsianos N Dohrn A El-Refai DA Falci SGM 24 39 10 26 2 3 29 12 21 24 10 30 29 15 23 10 20 Frentzen M Goddard G Hassan MM Hassan MM Hong T Hwaiti H Hwaiti H Jing M Joger A Khabaz BA Madfa A Madfa AA Madfa AA Meister J Merzouk N Nassif MS 6 5 21 8 11 Noman NA Shamala A Shamalah A Shamalah A Zaitone SA 2 3 18 10 Zaitone SA Al-Anesi MS Aldhorae K Aldhorae K 11 12 13 21 22 Aldhorae KA Al-Hadad Alhajj WA Alhajj WA Alkadasi BA 5 14 9 2 Alkulaibi MM Al-Marebi KG Almaweri AA Almaweri AA 3 27 26 25 30 29 6 20 23 1 7 4 17 24 8 28 16 19 Al-Moraissi E Al-Moraissi E Al-Moraissi E Alsameai AM Alsanabani F Alshaibani DAQ Alsubari RA Altawili ZM Al-Yousefi AA Al-Zendani EA Amran AN Baraja MM Bin Z Helmi JJ Mahgoub NM Mutahar M Mutahar NJ Yehia LA 15 39 INDEX - Subject Acupuncture 26 Anterior repositioning splint 25 Apical periodontitis 23 Arthrocentesis 27 Arthrogneous TMDs 25 Arthroscopy 27 Artificial saliva 20 Bonding 10 Canines 4 Caucasian 1 Close reduction 17 Composite 29 Corticosteroid 7 Dental education 18 Dental fluorosis 8 Dental practitioners 23 Dental students 11 Dental students 18 Dentist 11 Dentofacial 11 Dry needling 26 Endodontic treatment 18 Erosion 16 Fractures 19 Full soft stabilization splint 25 Human enamel 16 Hyaluronic acid 27 Incisors 19 Inferior alveolar nerve 9 Interceptive orthodontic 13 Intra-maxillary fixation 17 Ions 16 iRoot BP 28 iRoot BP Plus 28 Khat 5 Khat chewing 21 Knowledge 23 Laser etching 8 Laser etching 10 Laypersons 11 Leishmaniasis 14 Lower third surgery 7 Lupoid 14 Malocclusion 15 Mandibular condyle fractures 24 Mandibular third molars 24 Melanin 22 Mice 2 Microhardness 6 Minimal invasive 29 Mouthwash 20 Myofascial pain 26 Myogenus TMDs 25 Nanoparticles 2 Nanoparticles 3 Nerve lateralization 9 Ni-Cr alloys 20 Occlusal support 24 Open reduction 17 Oral serratiopetidase 7 Orthodontic brackets 8 Orthodontic brackets 10 Orthodontic treatment need 13 Pellicle 16 Perception 11 Physical therapy 27 Plaque form 14 Platelet-rich plasma 27 Postoperative sequelae 7 Prevalence 15 Primary dentition 15 Proteins 16 Pulp capping 28 Radiographic 4 Rat 3 Repair 29 Retrospective 4 Risk 22 Risk factors 21 Screw 17 SDS-PAGE 16 Shammah 5 Shear bond strength 8 Shear bond strength 10 Smoking 5 Smoking 21 Smoking 22 Step height 16 Temporomandibular disorders 26 Third molar 4 TMJ pain 27 Toxicity 2 Toxicity 3 Veneer 29 Water sorption 6 Yemen 4 Yemen 5 Yemen 13 Yemen 14 Yemen 22 40 TUIDC 1 Thamar University International Dental Conference INDEX - Subject Yemeni 21 Yemeni Norms 1 ZrO2 nanoparticles 6 Acid etching 8 Acid etching 10 Amalgam 29 Apical abscess 23 Cephalometric X-ray 1 Chymotrpsin 7 Conservative treatments 27 Corrosion resistance 20 Cuspid 30 Dental anomalies 15 Dental Implantation 30 Dental implants 9 Dental prosthesis 30 Endosequence root repair material 28 Endosseous 30 Esthetics 11 Full hard stabilization splint 25 Glass-ionomer 6 Impaction 4 Implant-supported 30 Khat 22 Mandibular angle fractures 24 Mandibular fracture 17 Nasal 14 Occlusal discrepancies 13 Orthodontic patients 12 Panoramic 12 Periodontal disease 21 Radiography 12 Root filling quality 18 Saliva 16 Silver 2 Silver 3 Tooth abnormalities 12 Tooth loss 5 Traumatic dental injuries 19 Wet needling therapy 26 41	42,685
https://openalex.org/W2982615270	OpenAlex	Open Science	CC-By	2,019	Performance of Lipoarabinomannan Assay using Cerebrospinal fluid for the diagnosis of Tuberculous meningitis among HIV patients	Richard Kwizera	Latin	Spoken	7,447	16,308	Abstract The diagnostic utility of the Background: Mycobacteria tuberculosis lipoarabinomannan (TB-LAM) antigen lateral flow assay on cerebrospinal fluid (CSF) for the diagnosis of tuberculous meningitis (TBM) has not been extensively studied and the few published studies have conflicting results. Lumbar CSF from 59 HIV-positive patients with suspected TBM Methods: was tested with TB-LAM and Xpert MTB/Rif Ultra. The diagnostic performance of CSF TB-LAM was compared to positive CSF Xpert MTB/Rif Ultra (definite TBM) and a composite reference of probable or definite TBM according to the uniform case definition. The diagnostic utility of the Background: Mycobacteria tuberculosis lipoarabinomannan (TB-LAM) antigen lateral flow assay on cerebrospinal fluid (CSF) for the diagnosis of tuberculous meningitis (TBM) has not been extensively studied and the few published studies have conflicting results. Lumbar CSF from 59 HIV-positive patients with suspected TBM Methods: was tested with TB-LAM and Xpert MTB/Rif Ultra. The diagnostic performance of CSF TB-LAM was compared to positive CSF Xpert MTB/Rif Ultra (definite TBM) and a composite reference of probable or definite TBM according to the uniform case definition. 1 Of 59 subjects, 12 (20%) had definite TBM and five (9%) had Results: probable TBM. With reference to definite TBM, CSF TB-LAM assay had a diagnostic sensitivity of 33% and specificity of 96%. When compared to a composite reference of definite or probable TBM, the sensitivity was 24% and specificity was 95%. There were two false positive tests with TB-LAM (3+ grade). In-hospital mortality in CSF TB-LAM positive patients was 17% compared to 0% in those with definite TBM by Xpert MTB/Rif Ultra but negative LAM. Lumbar CSF TB-LAM has a poor performance in diagnosing Conclusions: TBM. Both urine TB-LAM and Xpert Ultra should be further investigated in the diagnosis of TBM. Any reports and responses or comments on the article can be found at the end of the article. Keywords 1 1-3 4 1 1 5 1,5 1,5 5 1,4,5 1 2 3 4 5 Reviewer Status Invited Reviewers version 2 published 30 Sep 2019 version 1 published 19 Aug 2019 1 2 report report , Institute of Research for Maryline Bonnet Development (IRD), Montpellier, France 1 , University of KwaZulu-Natal Vinod Patel (UKZN), Durban, South Africa 2 19 Aug 2019, :123 ( First published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.1 30 Sep 2019, :123 ( Latest published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.2 v2 Open Peer Review Reviewer Status Invited Reviewers version 2 published 30 Sep 2019 version 1 published 19 Aug 2019 1 2 report report 19 Aug 2019, :123 ( First published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.1 30 Sep 2019, :123 ( Latest published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.2 v2 19 Aug 2019, :123 ( First published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.1 30 Sep 2019, :123 ( Latest published: 4 ) https://doi.org/10.12688/wellcomeopenres.15389.2 v2 Tuberculous meningitis, extra-pulmonary TB, lipoarabinomannan, TB-LAM, Xpert MTB/Rif Ultra, HIV, Diagnostics, cerebrospinal fluid Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Open Peer Review Any reports and responses or comments on the article can be found at the end of the article. RESEARCH NOTE Performance of Lipoarabinomannan Assay using Cerebrospinal fluid for the diagnosis of Tuberculous meningitis among HIV patients [version 2; peer review: 2 approved] Richard Kwizera , Fiona V. Cresswell , Gerald Mugumya , Micheal Okirwoth , Enock Kagimu , Ananta S. Bangdiwala , Darlisha A. Williams , Joshua Rhein , David R. Boulware , David B. Meya1,4,5 Infectious Diseases Institute, College of Health Sciences, Makerere University, Kampala, Uganda Clinical Research Department, London School of Hygiene and Tropical Medicine, London, WC1E 7HT, UK MRC-UVRI and LSHTM Uganda Research Unit, Entebbe, Uganda Department of Medicine, School of Medicine, College of Health Sciences, Makerere University, Kampala, Uganda Division of Infectious Diseases and International Medicine, Department of Medicine, University of Minnesota, Minneapolis, MN, 55455, USA Abstract The diagnostic utility of the Background: Mycobacteria tuberculosis lipoarabinomannan (TB-LAM) antigen lateral flow assay on cerebrospinal fluid (CSF) for the diagnosis of tuberculous meningitis (TBM) has not been extensively studied and the few published studies have conflicting results. Lumbar CSF from 59 HIV-positive patients with suspected TBM Methods: was tested with TB-LAM and Xpert MTB/Rif Ultra. The diagnostic performance of CSF TB-LAM was compared to positive CSF Xpert MTB/Rif Ultra (definite TBM) and a composite reference of probable or definite TBM according to the uniform case definition. Of 59 subjects, 12 (20%) had definite TBM and five (9%) had Results: probable TBM. With reference to definite TBM, CSF TB-LAM assay had a diagnostic sensitivity of 33% and specificity of 96%. When compared to a composite reference of definite or probable TBM, the sensitivity was 24% and specificity was 95%. There were two false positive tests with TB-LAM (3+ grade). In-hospital mortality in CSF TB-LAM positive patients was 17% compared to 0% in those with definite TBM by Xpert MTB/Rif Ultra but negative LAM. Lumbar CSF TB-LAM has a poor performance in diagnosing Conclusions: TBM. Both urine TB-LAM and Xpert Ultra should be further investigated in the diagnosis of TBM. Keywords Tuberculous meningitis, extra-pulmonary TB, lipoarabinomannan, TB-LAM, Xpert MTB/Rif Ultra, HIV, Diagnostics, cerebrospinal fluid Page 1 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Richard Kwizera ( ) Corresponding author: kwizerarichard@ymail.com : Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Writing – Original Author roles: Kwizera R Draft Preparation, Writing – Review & Editing; : Conceptualization, Formal Analysis, Funding Acquisition, Methodology, Project Cresswell FV Administration, Resources, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing; : Formal Analysis, Mugumya G Investigation, Validation, Writing – Review & Editing; : Formal Analysis, Investigation, Methodology, Writing – Review & Editing; Okirwoth M : Data Curation, Investigation, Methodology, Writing – Review & Editing; : Data Curation, Formal Analysis, Methodology, Kagimu E Bangdiwala AS Validation, Writing – Review & Editing; : Methodology, Project Administration, Resources, Supervision, Writing – Review & Editing; Williams DA : Project Administration, Resources, Supervision, Validation, Writing – Review & Editing; : Funding Acquisition, Project Rhein J Boulware DR Administration, Resources, Supervision, Validation, Writing – Review & Editing; : Funding Acquisition, Project Administration, Resources, Meya DB Supervision, Validation, Writing – Review & Editing No competing interests were disclosed. Competing interests: This work was supported by the Wellcome Trust [107742, 107743 and 210772]. This research was also supported in part by Grant information: the National Institute of Neurologic Diseases and Stroke (NINDS) and Fogarty International Center [R01NS086312, K01TW010268]. DBM and RK are currently supported through the DELTAS Africa Initiative grant [DEL-15-011] to THRiVE-2, from Wellcome Trust grant [107742] and the UK government. FVC is supported through a Wellcome Clinical PhD Fellowship [210772]. FVC is an honorary fellow of the Makerere University – UVRI Centre of Excellence for Infection and Immunity Research and Training (MUII-plus). MUII-plus is supported through the DELTAS Africa Initiative [107743]. The DELTAS Africa Initiative is an independent funding scheme of the African Academy of Sciences (AAS), Alliance for Accelerating Excellence in Science in Africa (AESA) and supported by the New Partnership for Africa's Development Planning and Coordinating Agency (NEPAD) with funding from the Wellcome Trust [107743] and the UK Government. The MRC/UVRI and LSHTM Uganda Research Unit is jointly funded by the UK Medical Research Council (MRC) and the UK Department for International Development (DFID) under the MRC/DFID Concordat agreement and is also part of the EDCTP2 programme supported by the European Union. Keywords p p g pp y p e funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. © 2019 Kwizera R . This is an open access article distributed under the terms of the , which Copyright: et al Creative Commons Attribution License permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Kwizera R, Cresswell FV, Mugumya G How to cite this article: et al. Performance of Lipoarabinomannan Assay using Cerebrospinal fluid Wellcome Open Research 2019, for the diagnosis of Tuberculous meningitis among HIV patients [version 2; peer review: 2 approved] 4 :123 ( ) https://doi.org/10.12688/wellcomeopenres.15389.2 Kwizera R, Cresswell FV, Mugumya G How to cite this article: et al. Performance of Lipoarabinomannan Assay using Cerebrospinal fluid Wellcome Open Research 2019, for the diagnosis of Tuberculous meningitis among HIV patients [version 2; peer review: 2 approved] 4 :123 ( ) https://doi.org/10.12688/wellcomeopenres.15389.2 19 Aug 2019, :123 ( ) First published: 4 https://doi.org/10.12688/wellcomeopenres.15389.1 Page 2 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 that urine TB-LAM may help to reduce mortality and predict poor outcomes9,10. The WHO recently added the TB-LAM assay onto its essential diagnostic list and recommended TB-LAM in hospitalised HIV positive adults with signs and symptoms of TB11,12. However, there are conflicting results about TB-LAM assay sensitivity for TBM diagnosis13 in CSF. With reference to definite TBM, Cox et al. found a 75% sensitivity using CSF from the fourth ventricle in an autopsy cohort from 91 HIV-infected adults14. However, Bahr et al. had no positive TB-LAM tests using lumbar CSF from 67 HIV patients with meningitis13. In light of these results, and now that Xpert MTB/RIF Ultra is used instead of Xpert MTB/RIF, we aimed to further explore the utility of CSF TB-LAM test for the diagnosis of TBM among HIV-positive adults presenting with suspected meningitis. Amendments from Version 1 REVISED Based on the two reviewers’ comments to version 1, major changes were made to the conclusion and Table 1 while minor changes were made to the results section as seen in the tracked changes. We added the suggested reference in the discussion section. Table 1 was modified to present the characteristics of patients with diagnosis of definite, probable, possible and not-TBM. A sentence was added on cerebral imaging. We also added a sentence about the patients who were positive with urine LAM and those who were positive with urine Ultra. We added an explanation about the 17% of patients had unknown outcome. A limitation was added that TB culture was not used. However, we have found Ultra to be more sensitive than culture (70% versus 43% against definite/probable) for the diagnosis of TBM in our population (Bahr, Lancet ID, 2018). In the conclusion section, we have mentioned the Fuji LAM that is a new LAM POC test that has higher sensitivity in urine than the determine LAM POC and should also be evaluated for diagnosis of TB meningitis both in urine and CSF. Lastly, based on the second reviewer’s comments, we would like to say that a cellular CSF is common in this population with advanced HIV disease even in the setting of a confirmed TB or CM infection (Cresswell, Int J Infect Dis 2018). The not-TBM group scored <6 points on the uniform case definition (n=16) or had a confirmed alternative diagnosis (n=10 with CM). Methods Study setting and participants Study setting and participants Between April 2018 and June 2019, we assessed and performed diagnostic lumbar punctures on HIV-positive patients admitted to Mulago National Referral Hospital with suspected meningitis in Kampala, Uganda. Screening for TBM was performed cross-sectionally as part of the High Dose Rifampicin for Tuberculous Meningitis (RIFT) trial (ISRCTN registration number ISRCTN42218549, last updated 24/04/2018)15. Therefore, we did not calculate a sample size for the current study but included all participants that fit the screening criteria for the RIFT trial15. All included participants were HIV-infected adults (≥18 years old) who provided written informed consent by participant or surrogate, with a suspected diagnosis of TBM (meningitis symptoms, clinical signs of meningism). Demographic information and baseline characteris- tics for participants were collected through clinical reviews using customized meningitis screening case report forms approved by the relevant ethics committees (Mulago Hospital Research Ethics Committee, Uganda National Council of Science and Technology, and the University of Minnesota). Opening pressures for CSF were measured using a manometer, followed by standard microbiology analysis (CrAg, cell count, protein, glucose, lactate, culture). Any further responses from the reviewers can be found at the end of the article Introduction In many human immunodeficiency virus (HIV) endemic countries, tuberculous meningitis (TBM) is the second most common cause of adult meningitis after cryptococcal meningitis1, and accounts for 1–5% of all tuberculosis (TB) cases2. TBM is the most severe form of TB and causes substantial morbidity and mortality in children and immunocompromised adults3,4. HIV infection is known to increase the risk of death in patients with TBM, as does TBM stage at the time of treatment initiation2,5. As is the case in cryptococcosis, high-quality nursing care is a critical component in managing TBM patients6. Similarly, diagnosis of TBM is very challenging, especially in resource-limited settings where diagnosis relies on a combina- tion of clinical, radiological and laboratory findings. The World Health Organisation (WHO) recommends Xpert MTB/RIF Ultra for the diagnosis of TBM using cerebrospinal fluid (CSF). Culture has many limitations related to turnaround time and sensitivity, and also requires considerable infrastructure and costs7. Therefore, the development of early point of care diagnosis for TBM is a priority. Recent studies have demonstrated that the next generation Xpert MTB/RIF Ultra is the most sensitive diagnostic test in HIV-positive adults7. However, Xpert MTB/RIF Ultra is not a bedside test, and thus access to same day results remain a challenge in many settings7. Diagnostic tests Diagnostic tests In addition to standard microbiology analysis, CSF was tested with TB-LAM (Alere, Massachusetts, USA), and the test strip interpreted as per manufacturer’s instructions. Briefly, the protective foil cover was removed from each test and the strip labelled with the participant’s number. Two drops (or 60μL) of CSF were added to the sample pad. The test was then read after 25 minutes under standard indoor lighting conditions. The reference card was used in interpretation of the results by holding it alongside the patient window. For positive results, purple/gray bars appeared in both the control window and the patient window of the strip. For negative results, one purple/ gray bar appeared in the control window of the strip and no bar appeared in the patient window of the strip. If there was no bar Assays based on the detection of mycobacterial lipoarabinoman- nan (TB-LAM) antigen in urine have emerged as potential point-of-care tests for extra-pulmonary TB8. There is evidence Page 3 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 in the control window of the strip, the result was considered invalid and the test repeated. The strips were retained and cross checked by a second researcher to corroborate the finding. receiving antiretroviral therapy at diagnosis. Among partici- pants reporting a headache (n=57), the median duration of headache was 14 days (IQR: 14, 24). The CSF opening pressures at baseline (n=45) had a median of 200 mmH2O (IQR: 120, 260). Overall, 55% (n=36) had an acellular CSF, whilst those with a CSF lymphocytic pleocytosis had a median CSF white blood cell of 160 cells/μL (IQR: 135, 268) (Table 1). Only about 10% of the participants had cerebral imag- ing done as the CT scanner was dysfunctional for part of the study period. About twenty five percent of the patients had a positive TB-LAM while 20% had a positive urine MTB/Rif Ultra. CSF was also tested with Xpert MTB/Rif Ultra (Cepheid). Briefly, 2ml of sample reagent was added to 1ml of whole CSF and then left to stand at room temperature for 15 minutes. Then, 2ml of the sample mixture was transferred into the Xpert MTB/Rif Ultra cartridge and loaded into the Xpert machine. The test was run for 90 minutes and results from the assay indicate whether or not Mycobacteria TB (MTB) was detected in the sample. Diagnostic tests If MTB was detected, the results also stated whether resistance to rifampin was detected. With respect to the reference standard of definite TBM (positive CSF Xpert TB/Rif Ultra), the CSF TB-LAM assay had a sensitivity of 33% (4/12), specificity of 96% (45/47), positive predictive value (PPV) of 67% (4/6), and negative predictive value (NPV) of 85% (45/53). When compared to a composite reference of definite/probable TBM, the TB-LAM assay had a sensitivity of 24% (4/17), specificity of 95% (40/42), PPV of 67% (4/6), NPV of 76% (40/53) (Table 2). There were two false positive tests with TB-LAM (3+ grade), without any CSF pleocytosis, normal protein, normal glucose, negative cryptococcal antigen, and normal CSF opening pressure. One patient was discharged alive without TB therapy. The second patient had a headache for 60 days at presentation, but they were lost to follow up (i.e. self-discharged) without an etiologic diagnosis. In-hospital mortality in CSF TB-LAM positive patients was 17% (1/6) compared to 0% (0/8) in those with definite TBM by Xpert MTB/Rif Ultra but negative LAM. About 17% of patients had unknown outcome. This was because the study population included patients screened for a clinical trial but only a minority were subsequently enrolled into the trial. We endeavoured to follow screen failures through to hospital discharge but this was not possible in all cases. Conclusion In conclusion, a rapid CSF point of care test for TBM is needed; however, this study demonstrated a poor diagnostic performance of the existing Alere TB-LAM on CSF among HIV-associated tuberculous meningitis. Our results corroborate the findings of a recent Zambian study which demonstrated 22% sensitiv- ity for CSF LAM against a reference standard of TB culture18. While the relatively modest sample size is a limitation, a larger sample size is unlikely to fundamentally alter the find- ings of sensitivity. One explanation could be that TB-LAM is likely not be found in sufficient quantities in lumbar CSF. TB culture was not used, which is also a limitation of the accuracy analysis. However, Xpert Ultra has a sensitivity that is greater than culture in our setting7. The novel Fujifilm SILVAMP TB-LAM (FujiLAM) assay has been shown to have higher sensitivity in urine than the Alere TB-LAM and Test analysis Data were analyzed using STATA version 14 (STATA, College Station, Texas). The disease prevalence, sensitivity, specificity, positive predictive values, negative predictive values and test accuracy were estimated at 95% confidence interval (CI). The diagnostic performance of CSF TB-LAM was compared to positive CSF Xpert MTB/Rif Ultra (definite TBM) and a composite reference of probable or definite TBM according to the uniform case definition16. Summaries were made in frequency & percentages for each baseline characteristic con- sidered as a categorical, and medians (interquartile range) when each characteristic is considered as a continuous variable. For baseline variables with some missing data, we calculated the statistics using the available numbers. Ethical statement Institutional review board approvals for the study and the associated screening process were obtained locally in Uganda (Mulago Hospital Research Ethics Committee, approval number MREC 1260); and from the London School of Hygiene and Tropical Medicine, UK (14388), University of Minnesota (1304M31361) and by the Uganda National Council of Science and Technology (HS136ES). Written informed consent for participation in the study and data publication was obtained from all participants or from their surrogates (e.g. family member or guardian) where the patient had altered mental status and did not have the capacity to provide consent. Results Overall, 59 HIV-positive hospitalized participants with suspected meningitis underwent diagnostic lumbar punctures, of which 20% (12/59) had definite TBM, 9% (5/59) had probable TBM, 25% (15/59) had possible TBM, and 46% (27/59) had not-TBM17. Of those with not-TBM (n=27), 10 had cryptococcosis. Women comprised 50% of participants with an overall median age for all participants of 33 years (interquar- tile range [IQR]: 28, 40). Only 29% of the participants were Page 4 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Table 1. Characteristics of the study population. Results Data presented are percentages (%), medians and interquartile ranges (IQR). N= number of participants with data for each parameter. * Participants with data available. ART = antiretroviral therapy, CSF = cerebrospinal fluid, WBC = white blood cells. Table 2. Summary of diagnostic performance of cerebrospinal fluid mycobacterial lipoarabinomannan assay for tuberculous meningitis. Table 2. Summary of diagnostic performance of cerebrospinal fluid mycobacterial lipoarabinomannan assay for tuberculous meningitis. Reference standard Disease prevalence Sensitivity Specificity PPV NPV Test Accuracy Definite/probable TBM 28.8% (17/59) 23.5% (4/17) 95.2% (40/42) 66.7% (4/6) 75.5% (40/53) 74.6% (44/59) 95% CI 17.8 to 42.1% 6.8 to 49.9% 83.8 to 99.4% 28.7 to 90.8% 70.1 to 80.2% 61.6 to 85% Definite TBM 20.3% (12/59) 33.3% (4/12) 95.7% (45/47) 66.7% (4/6) 84.9% (45/53) 83.1% (49/59) 95% CI 10.9 to 32.8% 9.9 to 65.1% 85.5 to 99.5% 29.3 to 90.6% 78.9 to 89.4% 71 to 91.6% Data presented are the percentage, numerator/denominator, and 95% confidence intervals (CI). Test Accuracy = overall probability that a patient will be correctly classified. PPV = Positive predictive value, NPV = negative predictive value, TBM = tuberculous meningitis. warrants evaluation for diagnosis of TB meningitis both in urine and CSF19. Data availability Underlying data Figshare: CSFLAM_data set revised.xlsx. https://doi.org/10.6084/ m9.figshare.9415853.v117 Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). Acknowledgements We thank institutional support from the IDI research office. Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). warrants evaluation for diagnosis of TB meningitis both in urine and CSF19. Results Baseline characteristics Overall (N=59) Definite TBM (n=12) Probable TBM (n=5) Possible TBM (n=15) Not-TBM (n=27) N* Statistic N* Statistic N* Statistic N* Statistic N* Statistic Women, n (%) 58 29 (50) 12 6 (50) 5 3 (60) 15 4 (26.7) 26 16 (61.5) Age in years, median (IQR) 58 33 (28-40) 12 29 (28-33) 5 26 (24-34) 15 35 (32-43) 26 34 (26-46) On ART, n (%) 47 29 (62) 11 7 (63.6) 2 2 (100) 11 3 (27.3) 23 17 (73.9) Headache, n (%) 57 46 (81) 12 10 (83.3) 5 4 (80) 14 12 (85.7) 26 20 (76.9) Duration of headache, median (IQR) days 45 14 (14-24) 10 17.5 (14-30) 4 14 (10.5-17.5) 12 17.5 (14-31.5) 19 14 (7-30) Glasgow Coma Scale score, mean (SD) 55 13 (2.6) 12 12.5 (2.9) 5 11.8 (2.4) 14 12.7 (2.9) 24 14.3 (2.1) CSF CrAg positive, n (%) 58 10 (17) 12 0 (0) 5 0 (0) 15 0 (0) 26 10 (38.4) CSF Opening Pressure, median (IQR) mmH2o 45 200 (120-260) 7 180 (70-240) 3 260 (95-400) 13 190 (120-270) 22 215 (120-260) Acellular CSF, n (%) 55 36 (55) 11 3 (27.3) 5 1 (20) 14 11 (78.6) 25 21 (84) CSF WBC in those with CSF WBC pleocytosis, median (IQR) cells/μL 55 160 (135-268) 8 280 (162.5-575) 4 173 (130-237.5) 3 80 (35-160) 4 145 (87.5-210) CSF protein, median (IQR) mg/dL 52 57 (28-141) 11 184 (107-316) 5 158 (147-215) 13 44 (35-72) 23 31 (22-61) CSF glucose, median (IQR) mg/dL 32 65 (34-82) 7 44 (19.8-61) 3 90 (68-108) 8 86 (56.3-104) 14 61 (31-80) CSF lactate, median (IQR) mmol/L 36 3.9 (2.2-9) 8 9.7 (8.2-11.2) 4 9.2 (6.3-11.1) 8 3.4 (2.3-8.1) 16 2.4 (1.9-3.8) Duration of hospitalization, median (IQR) days 46 7 (4-14) 9 11 (9-14) 4 14.5 (10-16.5) 9 4 (4-14) 24 5 (2-12.5) Status at discharge Alive, n (%) Dead, n (%) Unknown, n (%) 59 40 (68) 9 (15) 10 (17) 12 8 (66.7) 3 (25) 1 (8.3) 5 2 (40) 1 (20) 2 (40) 15 8 (53.3) 2 (13.3) 5 (33.3) 27 22 (81.5) 3 (11.1) 2 (7.4) Data presented are percentages (%), medians and interquartile ranges (IQR). N= number of participants with data for each parameter. * Participants with data available. ART = antiretroviral therapy, CSF = cerebrospinal fluid, WBC = white blood cells. Data availability Underlying data Acknowledgements We thank institutional support from the IDI research office. Page 5 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 References randomised controlled trial. Lancet. 2018; 392(10144): 292–301. PubMed Abstract \| Publisher Full Text \| Free Full Text randomised controlled trial. Lancet. 2018; 392(10144): 292–301. PubMed Abstract \| Publisher Full Text \| Free Full Text 11. WHO: The use of lateral flow urine lipoarabinomannan assay (LF-LAM) for the diagnosis and screening of active tuberculosis in people living with HIV. Geneva, Switzerland. 2015. Reference Source 2. Thao LTP, Heemskerk AD, Geskus RB, et al.: Prognostic Models for 9-Month Mortality in Tuberculous Meningitis. Clin Infect Dis. 2018; 66(4): 523–32. 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Gupta-Wright A, Corbett EL, Wilson D, et al.: Risk score for predicting mortality including urine lipoarabinomannan detection in hospital inpatients with HIV-associated tuberculosis in sub-Saharan Africa: Derivation and external validation cohort study. PLoS Med. 2019; 16(4): e1002776. PubMed Abstract \| Publisher Full Text \| Free Full Text 19. Broger T, Sossen B, du Toit E, et al.: Novel lipoarabinomannan point-of-care tuberculosis test for people with HIV: a diagnostic accuracy study. Lancet Infect Dis. 2019; 19(8): 852–61. PubMed Abstract \| Publisher Full Text \| Free Full Text 10. Gupta-Wright A, Corbett EL, van Oosterhout JJ, et al.: Rapid urine-based screening for tuberculosis in HIV-positive patients admitted to hospital in Africa (STAMP): a pragmatic, multicentre, parallel-group, double-blind, 10. Gupta-Wright A, Corbett EL, van Oosterhout JJ, et al.: Rapid urine-based screening for tuberculosis in HIV-positive patients admitted to hospital in Africa (STAMP): a pragmatic, multicentre, parallel-group, double-blind, Page 6 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Open Peer Review Current Peer Review Status: Version 2 02 October 2019 Reviewer Report © 2019 Bonnet M. This is an open access peer review report distributed under the terms of the Creative Commons , which permits unrestricted use, distribution, and reproduction in any medium, provided the original Attribution License work is properly cited. No competing interests were disclosed. Competing Interests: Reviewer Expertise: Clinical research on tuberculosis. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Version 1 16 September 2019 Reviewer Report https://doi.org/10.21956/wellcomeopenres.16813.r36425 © 2019 Patel V. This is an open access peer review report distributed under the terms of the Creative Commons , which permits unrestricted use, distribution, and reproduction in any medium, provided the original Attribution License work is properly cited. Vinod Patel Department of Neurology, Nelson R. Mandela School of Medicine, University of KwaZulu-Natal (UKZN), Durban, South Africa Department of Neurology, Nelson R. Mandela School of Medicine, University of KwaZulu-Natal (UKZN), Durban, South Africa The article is well written with no major flaws. Regarding the non-TBM category, although 10 patient had cryptococcal meningitis, detail regarding the non-TBM diagnoses is important as this is a control group and a patient without a clinical meningitis may allow for better specificity. I note that some CSF’s were acellular, what were these diagnoses? Page 7 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Please provide detail (a table with confirmatory findings for TB such as CXR, abdominal ultrasound findings, CSF findings etc.) regarding the aspects considered to arrive at a diagnosis of probable an possible TBM. A similar consideration for possible TBM. This would add clarity on the reliability of th outcomes. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes No competing interests were disclosed. Competing Interests: Reviewer Expertise: Assessing novel tests in the diagnosis of tuberculous meningitis. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Please provide detail (a table with confirmatory findings for TB such as CXR, abdominal ultrasound, scan findings, CSF findings etc.) regarding the aspects considered to arrive at a diagnosis of probable and possible TBM. A similar consideration for possible TBM. No competing interests were disclosed. Competing Interests: This would add clarity on the reliability of the test outcomes. No competing interests were disclosed. Competing Interests: No competing interests were disclosed. Competing Interests: Reviewer Expertise: Assessing novel tests in the diagnosis of tuberculous meningitis. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Author Response 26 Sep 2019 , College of Health Sciences, Makerere University, Kampala, Uganda Richard Kwizera , College of Health Sciences, Makerere University, Kampala, Uganda Richard Kwizera The article is well written with no major flaws. Thank you. Response: The article is well written with no major flaws. Thank you. Response: Regarding the non-TBM category, although 10 patient had cryptococcal meningitis, detail regarding the non-TBM diagnoses is important as this is a control group and a patient without a clinical meningitis may allow for better specificity. I note that some CSF’s were acellular, what were these diagnoses? Please provide detail (a table with confirmatory findings for TB such as CXR, abdominal ultrasound, scan findings, CSF findings etc.) regarding the aspects considered to arrive at a diagnosis of probable and possible TBM. A similar consideration for possible TBM. This would add clarity on the reliability of the test outcomes. y y A cellular CSF is common in this population with advanced HIV disease even in the Response: setting of a confirmed TB or CM infection (Cresswell, Int J Infect Dis 2018). The not-TBM group scored <6 points on the uniform case definition (n=16) or had a confirmed alternative diagnosis (n=10 with CM). Page 8 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Maryline Bonnet Institute of Research for Development (IRD), Montpellier, France This manuscript reports the results of a nested study in a large clinical trial, evaluating the accuracy of the CSF LAM for diagnosis of tuberculosis meningitis using both definite TB and composite reference of probable and definite TB based on standard case definitions. The study reports low sensitivity and high specificity and concludes on the modest role of the CSF LAM for diagnosis of TB meningitis. These results are important and the manuscript is well written. No major comments. Minor comments: In the introduction, I suggest the authors adding the recent publication by Siddiqi .(2019) in their et al references. 1 Minor comments: In the introduction, I suggest the authors adding the recent publication by Siddiqi .(2019) in their et al references. 1 In Results, I would suggest the authors to present the characteristics of patients with diagnosis of probable TB using the uniform case definitions. I am surprised by the low proportion of probable TB as compared to definite TB. It would be interesting to know the score of patients with probable TB using the uniform case definition criteria. Were cerebral imaging criteria used? It would be also interesting to know the proportion of patients that fit the score of possible TB meningitis using the uniform case definitions. It is indeed important to know the proportion of possible TB meningitis cases based on the uniform cases definitions that were finally classified as non TB meningitis for the accuracy analysis. It would be interesting to know the proportion of patients with TB positive results from another specimen than CSF, which is also an important criteria for diagnosis of probable TB meningitis. One option could be to present the patients’ characteristics by definite TB, probable TB and others in Table 1. How do the authors explain that 17% of patients had unknown outcome of death or alive at discharge? It is quite high in a context of a nested study in a clinical trial. TB culture was not used, which is a limitation of the accuracy analysis. However, Xpert Ultra has a sensitivity that is very close to culture. This could be mentioned as a limitation. TB culture was not used, which is a limitation of the accuracy analysis. However, Xpert Ultra has a sensitivity that is very close to culture. This could be mentioned as a limitation. https://doi.org/10.21956/wellcomeopenres.16813.r36338 © 2019 Bonnet M. This is an open access peer review report distributed under the terms of the Creative Commons , which permits unrestricted use, distribution, and reproduction in any medium, provided the original Attribution License work is properly cited. No competing interests were disclosed. Competing Interests: No competing interests were disclosed. Competing Interests: 09 September 2019 Reviewer Report https://doi.org/10.21956/wellcomeopenres.16813.r36338 Maryline Bonnet It would be also very interesting to have the results of the urine LAM if used. In the study by Siddiqui et al. the urine LAM had higher sensitivity than the CSF LAM in patients with presumptive TB meningitis. It would be also very interesting to have the results of the urine LAM if used. In the study by Siddiqui et al. the urine LAM had higher sensitivity than the CSF LAM in patients with presumptive TB meningitis. In the conclusion, the authors could mention the Fuji LAM that is a new LAM POC test that has higher sensitivity in urine than the determine LAM POC and should also be evaluated for diagnosis of TB Page 9 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 sensitivity in urine than the determine LAM POC and should also be evaluated for diagnosis of TB meningitis both in urine and CSF. References 1. Siddiqi OK, Birbeck GL, Ghebremichael M, Mubanga E, Love S, Buback C, Kosloff B, Ayles H, Atadzhanov M, Dheda K, Koralnik IJ: Prospective Cohort Study on Performance of Cerebrospinal F (CSF) Xpert MTB/RIF, CSF Lipoarabinomannan (LAM) Lateral Flow Assay (LFA), and Urine LAM LF Diagnosis of Tuberculous Meningitis in Zambia. . 2019; (8). \| J Clin Microbiol 57 PubMed Abstract Publisher Full Text Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes No competing interests were disclosed. Competing Interests: Reviewer Expertise: Clinical research on tuberculosis. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Author Response 26 Sep 2019 , College of Health Sciences, Makerere University, Kampala, Uganda Richard Kwizera This manuscript reports the results of a nested study in a large clinical trial, evaluating the accura of the CSF LAM for diagnosis of tuberculosis meningitis using both definite TB and composite reference of probable and definite TB based on standard case definitions. sensitivity in urine than the determine LAM POC and should also be evaluated for diagnosis of TB meningitis both in urine and CSF. 1. Siddiqi OK, Birbeck GL, Ghebremichael M, Mubanga E, Love S, Buback C, Kosloff B, Ayles H, Atadzhanov M, Dheda K, Koralnik IJ: Prospective Cohort Study on Performance of Cerebrospinal Fluid (CSF) Xpert MTB/RIF, CSF Lipoarabinomannan (LAM) Lateral Flow Assay (LFA), and Urine LAM LFA for Diagnosis of Tuberculous Meningitis in Zambia. . 2019; (8). \| J Clin Microbiol 57 PubMed Abstract Publisher Full Text Maryline Bonnet The study reports low sensitivity and high specificity and concludes on the modest role of the CSF LAM for diagnosis o TB meningitis. These results are important and the manuscript is well written. Thank you Response: No major comments. Minor comments: In the introduction, I suggest the authors adding the recent publication by Siddiqi Comment: et a Wellcome Open Research 2019, 4:123 Last updated: 02 O Are the conclusions drawn adequately supported by the results? Yes : Thank you for this suggestion. This has been done in the modified Table 1. Response Comment: How do the authors explain that 17% of patients had unknown outcome of death or alive at discharge? It is quite high in a context of a nested study in a clinical trial. This study population included patients screened for a clinical trial but only a minority Response: were subsequently enrolled into the trial. We endeavoured to follow screen failures through to hospital discharge but this was not possible in all cases. Comment: TB culture was not used, which is a limitation of the accuracy analysis. However, Xpert Ultra has a sensitivity that is very close to culture. This could be mentioned as a limitation. We have found Ultra to be more sensitive than culture (70% versus 43% against Response: definite/probable) for the diagnosis of TBM in our population (Bahr, Lancet ID, 2018). However we agree addition to culture would have been positive so this has been added to the limitations. Comment: It would be also very interesting to have the results of the urine LAM if used In the study 1 Comment: One option could be to present the patients’ characteristics by definite TB, probable TB and others in Table 1. : Thank you for this suggestion. This has been done in the modified Table 1. Response Comment: How do the authors explain that 17% of patients had unknown outcome of death or alive at discharge? It is quite high in a context of a nested study in a clinical trial. This study population included patients screened for a clinical trial but only a minority Response: were subsequently enrolled into the trial. We endeavoured to follow screen failures through to hospital discharge but this was not possible in all cases. Comment: TB culture was not used, which is a limitation of the accuracy analysis. However, Xpert Ultra has a sensitivity that is very close to culture. This could be mentioned as a limitation. We have found Ultra to be more sensitive than culture (70% versus 43% against Response: definite/probable) for the diagnosis of TBM in our population (Bahr, Lancet ID, 2018). However we agree addition to culture would have been positive so this has been added to the limitations. Comment: It would be also very interesting to have the results of the urine LAM if used. In the study by Siddiqui . Are the conclusions drawn adequately supported by the results? Yes Author Response 26 Sep 2019 , College of Health Sciences, Makerere University, Kampala, Uganda Richard Kwizera This manuscript reports the results of a nested study in a large clinical trial, evaluating the accuracy of the CSF LAM for diagnosis of tuberculosis meningitis using both definite TB and composite reference of probable and definite TB based on standard case definitions. The study reports low sensitivity and high specificity and concludes on the modest role of the CSF LAM for diagnosis of TB meningitis. These results are important and the manuscript is well written. Thank you Response: No major comments. Minor comments: In the introduction, I suggest the authors adding the recent publication by Siddiqi Comment: et al .(2019) in their references. 1 Page 10 of In the introduction, I suggest the authors adding the recent publication by Siddiqi ent: et al Page 10 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 .(2019) in their references. We have now added this suggested reference in the discussion section. Response: Comment: In Results, I would suggest the authors to present the characteristics of patients with diagnosis of probable TB using the uniform case definitions. Table 1 has been modified to present the characteristics of patients with diagnosis of Response: definite, probable, possible and not-TBM. Comment: I am surprised by the low proportion of probable TB as compared to definite TB. This is likely to be because we used Ultra and not culture or Xpert as the reference Response: standard. We have found Ultra to be better at confirming TBM in our population (Bahr, Lancet ID, 2018) Comment: Were cerebral imaging criteria used? : Only about 10% (n/N) of the participants had cerebral imaging done as the CT scanner Response was dysfunctional for part of the study period. Comment: It would be also interesting to know the proportion of patients that fit the score of possible TB meningitis using the uniform case definitions. These have been added in Table 1. Response: Comment: It would be interesting to know the proportion of patients with TB positive results from another specimen than CSF, which is also an important criteria for diagnosis of probable TB meningitis. 25% of the patients were positive with urine LAM while 20% were positive with urine Response: Ultra. Comment: One option could be to present the patients’ characteristics by definite TB, probable TB and others in Table 1. Are the conclusions drawn adequately supported by the results? Yes the urine LAM had higher sensitivity than the CSF LAM in patients with et al presumptive TB meningitis. 25% of the patients were positive with urine TB-LAM LFA (Alere). The details of the Response: urine TB diagnostics are being analysed currently and will be presented separately. Page 11 of 12 Wellcome Open Research 2019, 4:123 Last updated: 02 OCT 2019 Comment: In the conclusion, the authors could mention the Fuji LAM that is a new LAM POC test that has higher sensitivity in urine than the determine LAM POC and should also be evaluated for diagnosis of TB meningitis both in urine and CSF. g g Thank you for this suggestion. This has been added in the conclusion. Response: No competing interests were disclosed. Competing Interests: Page 12 of 12	26,783
https://openalex.org/W2310062959	OpenAlex	Open Science	CC-By	2,000	Paletli ve Helezonik Karıştırıcılarda Yem Katkı Maddeleri Karışım Homojenliğinin Belirlenmesi	KOÇAK İlhami;ERDOĞAN	Turkish	Spoken	7,011	18,334	Giriş Ayn ı konuda Roche Ilaçlar ı A. Ş .'de yay ınlad ığı ürün katoloğunda, premiksin tan ı m ı , ideal premiks özellikleri ve premiks kalitesini etkileyen faktörler üzerinde durmu ştur. Yay ı nda ayr ıca vitamin ve minerallerin kar ışım içersindeki stabiliteleri ile di ğer aktif maddeler hak ı k ında da bilgi vermiştir (Anonymous 1997). Günümüzde üretilmekte olan karma yemlere, hayvanlar ı n ihtiyac ı olan, vitamin, mineral ve koruyucu özelliği olan baz ı maddeler kat ılmaktad ır. Ancak gereksinim duyulan bu maddeler yeme do ğrudan kat ı lmazIar. Bu aktif maddeler ön kar ışı m şeklinde ve yem katk ı maddeleri ad ı alt ında üretilip, yem fabrikalar ı na bu şekilde pazarlan ırlar. Yem katk ı maddelerinin kar ışı m ı konusunda Merc Animal Health şirketi yay ınlad ığı bültende; kar ışt ırma işleminin nas ıl yap ılmas ı gerekti ği, kar ışım düzgünlüğünün nas ıl sağlanaca ğı , kar ışım kriterlerinin neler oldu ğu, laboratuvar ortam ı n ı n bu konulara nas ı l destek oldu ğ u aç ı klanm ışt ı r. Kar ışt ı rı c ı çe şitlerinden yatay, dikey ve devirmeli kar ışt ı r ıc ılar hakk ında bilgi verilmi ştir (Anonymous 1980). Yem katk ı maddeleri (premiksler), ta şıy ıc ı ve aktif maddelerden olu şur. Ta şı y ıc ı maddeler, kalsiyum karbonat ve bitkisel kökenli maddeler olarak ikiye ayr ı l ı r. Ta şıy ıc ı maddeler yurt içinden rahatl ıkla karşılanabilirken, aktif maddelerin hemen hepsi yurt d ışından sağlanmaktadir. Yine kar ışım konusunda, Pfizer Ilaçlar ı A.Ş. premiks sektörü ile do ğrudan ili şki halinde olan yem sektöründe araştırmalar yapm ışt ır. Bu ara şt ırman ı n yer ald ığı yay ında doğru kar ışı m ı n nas ıl yap ılacağı , kar ışıma nelerin etkili olduğu anlat ı lm ışt ır. Yay ında helezonik kan şt ınc ı lar ile paletli kar ışt ı rıc ılar ın performans analizleri ile bu kan ştınc ılar hakk ında öneriler ve dikey mikserler hakk ında aç ıklay ıc ı bilgi verilmi ştir (Anonymous 1992). Yem katk ı maddelerinin tan ım ı ve içeriği konusunda bir çok ara şt ırmalar yap ı lm ışt ır. Özellikle vitamin ve premiks şirketleri bu konuya çok önem vermi şlerdir. DOI: 10.1501/Tarimbil_0000001002 DOI: 10.1501/Tarimbil_0000001002 TARIM BILIMLERI DERGISI 2000, 6 (4) 92-94 Geliş Tarihi : 04.07.2000 Geliş Tarihi : 04.07.2000 Özet: Bu araşt ı rmada, yatay (paletli ve helezonik) kan şt ı ncı larda yem katk ı maddeleri (Premiks) nin kar ışı m homojenliği incelenmiştir.Giriş bölümünde, yem katk ı maddelerinin özellikleri ve kan şt ı rı c ı tipleri aç ıklanm ışt ı r. Ayr ı ca, bu konuda yap ı lan çal ışmalar gözden geçirilmi ştir. Çal ışmada, paletli ve helezonik kar ışt ı r ıc ı larla denemeler yap ı lm ışt ı r.Bir premiks fabrikas ı nda yap ılmış denemelerden elde edilen kar ışı m örnekleri, analiz edilerek, homojenlik de ğerleri hesaplanm ış ve yorumlanm ışt ı r. Paletli karışt ı rıcı ile uygun homojenlik de ğerine daha k ısa sürede ula şı lm ışt ı r. Anahtar Kelimeler: Yem katk ı maddeleri (premiks), kar ışı m, homojenlik Yüksek Lisans Tezinden Özetlenmi ştir 1 Beypiliç - Bolu 2 Ankara Üniv. Ziraat Fak. Tar ı m Makinalar ı Bölümü - Ankara Paletli ve Helezonik Kar ışt ı rıc ılarda Yem Katk ı Maddeleri Karışım Homojenliğinin Belirlenmesi Ilhami KOÇAK' Doğan ERDOĞAN2 Determining of Homogenity of the Feed Grade Mixture in Mixers with Palette and Auger Abstract: The homogenity of the premix mixture in the horizontal (pallette and auger) mixers was studied in this survey. The Feed additive its characteristics and mixer types discussed at first. In addition, researchs in this field was summerized. The trails with the palette and auger mixers were realized at a premix firm. Then samples were taken and analyzed in laboratory. The values of homogeneity were calculated and interpreted according to the results of the analysis. To suitable homogenity was reached shorter period with pallette mixer than auger mixer. Key Words: Premix, mixture, homogeneity Key Words: Premix, mixture, homogeneity Giriş Bu şirketlerden biri olan Merck Animal Healt, yay ınlad ığı bültende; premiksin tan ı m ı ile birlikte, premiks yap ı m ında kullan ılan hamrr ıaddeler konusunda da aç ı kl ık getirmi ştir (Anonymous 1980). KOÇAK, I. ve D. ERDO ĞAN, "Paletli ve helezonik kari şt ıncilarda yem katk ı maddeleri kar ışı m homojenliğ inin belirlenmesi" KOÇAK, I. ve D. ERDO ĞAN, "Paletli ve helezonik kari şt ıncilarda yem katk ı maddeleri kar ışı m homojenliğ inin belirlenmesi" 93 Kar ışt ı rd ı çeşitleri Amerikada yay ınlanan, tar ım ve besin materyallerinin kar ışı m ı konusunun işlendiği yay ında da ele al ı nm ışt ı r (Niranjan, K., Smith, D. L. O. 1994). Kar ışt ı rd ı çeşitleri Amerikada yay ınlanan, tar ım ve besin materyallerinin kar ışı m ı konusunun işlendiği yay ında da ele al ı nm ışt ı r (Niranjan, K., Smith, D. L. O. 1994). kar ışı m zaman ı ve dizayn ı aç ıs ı ndan dikey, devirmeli ve pnömatik gibi kar ışt ı r ıc ılara göre daha avantajl ı bulunmuşlard ı r. Materyal ve Yöntem Premiks üretiminde en önemli nokta kar ışım ın homojen ç ıkmas ıd ır. Çünkü homojen olmayan kar ışım ın yem sektörü için hiçbir önemi yoktur. Homojenli ğin ne olduğu, niçin gerekli oldu ğu, nelerin etkili oldu ğu, hangi yöntemlerle ölçüldü ğü, nas ıl hesapland ığı , varyasyon katsay ıs ın ın ne oldu ğu konusunda yay ınlar bulunmaktad ı r. Karışım ı oluşturan hammaddelerin kar ışı m ın her bölgesinde eşit olarak saptanmas ı n ı homojenlik olarak belirten Merck Animal Health, ayr ıca karışım zaman ı n ın homojenlik üzerindeki etkisi hakk ında da bilgi vermi ştir (Anonymous 1980). Araşt ırmada, kapasiteleri 1 ton/h olan iki tip kar ışt ı r ıc ı kullan ılm ışt ır. Bunlardan helezonik kar ışt ır ıc ı n ın milinin dönme say ıs ı 48 min -1 dir. Iletim sistemi kay ış-kasnak olan kar ışt ı r ıc ı elektrik motoru ile çal ışt ı r ılmaktatd ı r. Karışt ı r ıc ı tek mil ancak çift şeritlidir. Paletti karişt ır ıc ı n ı n mil devir say ıs ı 32 min-1 dir. Mil say ıs ı iki olup elektrik motoru ile tahrik edilmektedir. Araşt ırmada deneme hammaddeteri olarak ta şıy ıc ı ve aktif maddeler kullan ı lm ışt ır. Ta şıy ıc ı madde olarak kullan ılan kalsiyum karbonat ın (CaCO3) tane büyüklü ğü 400 mm'dir. Aktif madde olarak kullan ı lan Virginamiycin % 100'lüktür. Homojenliğin geçmişi hakk ındaki bilgi, tar ım ve besin materyallerinin kar ışım ı konulu kaynakta incelenmi ştir (Niranjan, K., Smith, D. L. O, 1994). Buhler Brother Ltd. Şirketi yay ınlad ığı bültende de homojenlik hakk ında bilgi vermiştir (Tremer, D. 1995). Denemeler Istanbul il s ın ı rlar ı içindeki bir özel premiks haz ırlama fabrikas ı nda yap ı lmışt ır. Tesis yar ı otomatik olarak çal ışmaktad ı r. Istenilen kalite standard ına ulaşmak için gerekli karışt ırma zaman ı aç ıs ından çe şitli tip karışt ı rıc ı lar aras ında büyük farkl ı l ıklar vard ı r. Karışt ırma zaman ı , bir karışt ı r ıc ı tipinden diğerine 2-30 dakika aras ında değ işebilir. Bununla birlikte her kar ışt ı r ıc ın ın bir optimum kan şt ı rma zaman ı olduğuna dikkat edilmelidir. Çünkü belirli bir homojenlik derecesi elde edildikten sonra kar ışım daha fazla iyile ştirilemez (Anonymous 1996). Materyal ve Yöntem Araşt ı rmada kar ışt ı r ıc ı tipi ile homojenlik aras ındaki ilişkinin, deneysel olarak belirlenmesi planlanm ışt ı r. Paletli kan şt ı rıc ılarda 1, 3, 5, 7 ve 9. Dakikalarda, helezonik kar ışt ı r ıc ılarda ise 10, 20, 30, 35 ve 40 inci dakikalarda örnekler al ı nm ış ve laboratuvarda analiz edilmiştir. Elde edilen değerlerle analitik hesaplamalar yap ılm ış ve istatistiki olarak de ğerlendirilmi ştir. Sonuçlar çizelge ve grafiklerle verilmi ştir. Karışt ı r ıc ı tipinin homojenliğe etkisi büyüktür. Daha önce yap ılan ara şt ı rma sonuçlar ı nda, özellikle yatay kan şıt ı rc ılar üzerinde durulmu ştur. Yatay kar ışt ı rıc ılar, Kasnak Gövde Helezon mili Zincir disti Gövde /_ Palet mili Şekil 1.Helezonik yatay kar ışt ır ıc ı (Anonymous 1996) Şekil 2. Paletli yatay kar ışt ı r ıc ı (Anonymous 1996) Zincir disti Gövde /_ Palet mili Kasnak Gövde Helezon mili Kasnak Şekil 1.Helezonik yatay kar ışt ır ıc ı (Anonymous 1996) Şekil 2. Paletli yatay kar ışt ı r ıc ı (Anonymous 1996) 20 75 10 - 8 10 0 2 4 6 Süre (dakika) Şekil 3. Paletli karışt ı r ıcı da zamana göre CV de ğerleri 5- O 30 - 2 5 - > 20 - U 15 ı o 5 0 TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 0 20 40 60 80 Süre (dakika ) Şekil 4 Helezonik kar ışt ı r ıcda zamana göre CV değerleri TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 94 94 20 75 10 - 8 10 0 2 4 6 Süre (dakika) Şekil 3. Paletli karışt ı r ıcı da zamana göre CV de ğerleri 5- O , , y 20 75 10 - 8 10 0 2 4 6 Süre (dakika) 5- O Bulgular ve Tartışma Paletli ve helozonik karışt ı rıcı ile yap ılan denemelerde elde edilen aktif madde (Virginamycin) miktar ı PPm cinsinden çizelge 1 ve çizelge 2'de belirtilmiştir. Çizelge 1. Paletli kar ışt ı rıc ıda süreye göre, kar ışı m içerisindeki aktif madde miktarlar ı (PPm ve CV de ğerleri Süre (dakika) 1.örnek 2.örnek 3.örnek 4.örnek 5.örnek % CV 1. 50 70 57 45 78 22.9 3. 55 53 49 52 41 10,9 5. 42 48 43 47 45 5,6 7. 39 41 42 40 38 3,9 9. Treme, D. 1995. Symposium on Today's and Equip for the Prodction of compound Feed Swisstech Istanbul 88 March 1-4 Buhler - Miag. Buhler Brothers Ltd. CH-9240, Switzerland. Materyal ve Yöntem 45 50 65 55 60 14,3 Süre (dakika) Şekil 3. Paletli karışt ı r ıcı da zamana göre CV de ğerleri Çizelge 2. Helezonik kar ışt ı r ı c ı da süreye göre kar ışı m içersindeki aktif madde miktarlar ı (PPm) Çizelge 2. Helezonik kar ışt ı r ı c ı da süreye göre kar ışı m içersindeki aktif madde miktarlar ı (PPm) 30 - 2 5 - > 20 - U 15 ı o 5 0 0 20 40 60 80 Süre (dakika ) Şekil 4 Helezonik kar ışt ı r ıcda zamana göre CV değerleri 30 - 2 5 - > 20 - U 15 ı o 5 0 0 20 40 60 80 Süre (dakika) 1.örnek 2.örnek 3.örnek 4.örnek 5.örnek % CV 10. 60 70 130 115 125 32.5 20. 70 115 120 80 65 28.6 30. 80 90 100 95 60 18 40. 70 50 60 65 55 13.1 50. 50 55 50 55 40 12.1 60, 42 43 45 50 45 6.8 70. 50 55 53 55 62 8.8 Elde edilen değerlere göre çizilen grafikler şekil 3 ve şekil 4'de gösterilmi ştir. Denemelerde, her iki kar ışt ırıc ıda da ayn ı çe şit taşı y ıc ı madde ve aktif madde kullan ılmas ına karşı n, paletli kar ışt ı r ıc ıda uygun bir homojenlik de ğerine daha az zamanda ula şı lm ışt ır. Bundan dolay ıd ır ki art ık premiks sektöründe paletli kar ışt ı r ıc ılar tercih edilmektedir. Süre (dakika ) Şekil 4 Helezonik kar ışt ı r ıcda zamana göre CV değerleri Kaynaklar Niranjan, K., D. L. O. Smith, 1994. Mixing proces for agricultural and food Materials; Part 5 Review of mixer types. Journal of agricultural engeneer Research, 59, 145-161. Niranjan, K., D. L. O. Smith, 1994. Mixing proces for agricultural and food Materials; Part 5 Review of mixer types. Journal of agricultural engeneer Research, 59, 145-161. Niranjan, K., D. L. O. Smith, 1994. Mixing proces for agricultural and food Materials; Part 5 Review of mixer types. Journal of agricultural engeneer Research, 59, 145-161. Anonymous, 1980. Mixing microingredients in Feed Merck Animal Healt, Nevjersy. U.S.A. Anonymous, 1992. Yem üretimindeki eksik halka kaliteyi belirler. Pfizer Ilaçlar ı A. Ş . yay ı n ı , Istanbul. Anonymous, 1996. Günümüz yem fabrikalar ı . Pfizer ilaçlar ı yay ı n ı , Istanbul. Anonymous, 1997. Vademecum. Anonymous, 1980. Mixing microingredients in Feed Merck Animal Healt, Nevjersy. U.S.A. Anonymous, 1992. Yem üretimindeki eksik halka kaliteyi belirler. Pfizer Ilaçlar ı A. Ş . yay ı n ı , Istanbul. Anonymous, 1996. Günümüz yem fabrikalar ı . Pfizer ilaçlar ı yay ı n ı , Istanbul. Anonymous, 1997. Vademecum. Roche Ilaçlar ı A.Y. yay ı n ı , I t b l Niranjan, K., D. L. O. Smith, 1994. Mixing proces for agricultural and food Materials; Part 5 Review of mixer types. Journal of agricultural engeneer Research, 59, 145-161. Treme, D. 1995. Symposium on Today's and Equip for the Prodction of compound Feed Swisstech Istanbul 88 March 1-4 Buhler - Miag. Buhler Brothers Ltd. CH-9240, Switzerland. Niranjan, K., D. L. O. Smith, 1994. Mixing proces for agricultural and food Materials; Part 5 Review of mixer types. Journal of agricultural engeneer Research, 59, 145-161. Geliş Tarihi: 07.07.2000 Özet: Araşt ı rma, 1994 ve1995 y ı llarında Ankara koşullarında Atatürk Orman Çiftli ği ve Ankara Üniversitesi Ziraat Fakültesi Kenan Evren Ara şt ı rma ve Uygulama Çiftliği olmak üzere iki farkl ı lokasyonda yürütülmüştür. Araşt ı rman ı n amac ı, m ıs ır çeşitlerinde verim ve baz ı verim öğelerinin adaptasyon ve stabilite özelliklerini belirlemektir. Stabilite parametresi olarak; ortalama, regresyon katsay ıs ı (b), a değeri ve regresyondan sapma de ğeri kullan ı lm ışt ı r. Elde edilen sonuçlara gore; verim yönünden P-3751, P-3394 ve Franca çe şitleri diğer çeşitlere oranla tüm çevrelerde daha uyumlu ve stabil bulunmu ştur. Diğer verim öğeleri yönünden m ıs ı r çeşitlerinin gösterdiği uyum ve stabilitenin farkl ı olduğu anlaşı lm ıştır. Anahtar Kelimeler: Çeşit x çevre ilişkisi, stabilite parametreleri, m ı s ı r. Giriş melezlerin çevreye tepkilerinin tek melezlerden daha az olduğunu ve çift melezlerin stabilite performans ın ın tek melezlere gore daha iyi oldu ğunu aç ıklam ışlard ı r. Tarım ekosisemlerinde yeti ştirilen tüm bitkilerde olduğu gibi m ıs ır bitkisinin verimi de kalitatif bir karakter olup birçok faktörün etkisi alt ındad ır, Bu faktörler genotip ve çevre ko şulları olarak değerlendirilmektedir (Emeklier 1997). Farkl ı genotiplerin çevre ko şullar ına gösterdi ğ i tepki de değ işik olmaktad ır. Genetik bak ımdan saf olan çeşitler kötü çevre ko şullar ından yap ılar ındaki biyolojik kararl ı l ık nedeniyle daha az etkilenmektedir (Becker 1981). Eberhart ve Russell (1966), iki tek melez diallel m ıs ırda genetik farkl ı l ı klar ı incelemiş ve hatlar aras ındaki genetik farkl ı l ıklar ı çevre indeksi üzerindeki regresyonla belirlemiş, ve pek çok melezin regresyondan sapma karelerinin tahmininin s ıf ıra yak ın olduğunu bildirmişlerdir. Ara şt ı r ıc ılar, yağış dağı l ım ın ın önemli bir çevre faktörü olmas ı nedeniyle, erken ve geç ekim zamanlar ı n ın her bir lokasyonda ayr ı bir çevre olarak kullan ılabiteceğini belirtmişler, bunun gibi dü şük ve yüksek bitki popuiasyonu, orta ve yüksek gübreleme oran ı n ın belli bir" bölgede çevre ko şullar ın ı art ı rmak için kullan ı labileceğinr aç ıklam ışlard ı r. Islahç ı aç ıs ından önemli olan, bir bölge için geliştirilen yeni çeşidin o bölgenin kötü çevre ko şullar ında bile ortalama verimin alt ı na dü şmeyecek, iyi koşullarda ise en yüksek verimi verecek gücü stabii alarak göstere bilmesidir (Özgen 1994). Genotiplerin stabilitesini aç ıklamada kullan ılan ortalama değerin zamanla yetersiz olduğunun anlaşılmas ıyla, çeşit x çevre etkile şiminde genotipin pay ın ı belirlemek için de ğ işik yöntemler geliştirilmi ştir, Çe şit x çevre etkile şiminin regresyon analizi kullan ılarak belirlenebileceği ilk kez 1938 y ı l ında Yates ve Cochran taraf ından aç ıklanm ışt ır ( Francis ve Kannenberg 1978). Francis ve Kannenberg (1978), onbe ş tekmelez m ıs ır ı onalt ı farkl ı çevrede yeti ştirerek elde ettikleri verilere değ işik istatistik yöntemlerini uygulam ışlar, genotipleri verim stabilitesine gore grupland ırmada; verim ortalamas ı ve varyasyon katsay ıs ın ın önemli olduğunu belirtmi şlerdir. Ankara Only. Ziraat Fak. Tarla Bitkileri Bölümü -Ankara Stability Analysis and Adaptation of Yield and Some Yield Components in Maize Abstract: The research was conducted in Ankara environments in the fields of Atatürk Orman Çiftli ği and Ankara University, Faculty of Agriculture, Research and Application Farm in the years of 1994 and1995.The objective of this research was to determine adaptation and stability parameters of yield and yield components in maize cultivars. Mean value ( X ), regression coefficient (b), regression line intercept (a) and deviation from regression( S 2 d ) were used as adaptation and stability statistic. According to results, the yields of cultivars, P-3751, P-3394 and Franca were more adaptable and stable than the other genotypes. Stability and adaptability of genotypes as regards of other yield components were different than each others. Key Words: Genotype x environment interaction, stability parameters, maize. Materyal ve Yöntem Roche Ilaçlar ı A.Y. yay ı n ı , Istanbul. Anonymous, 1997. Vademecum. Roche Ilaçlar ı A.Y. yay ı n ı , Istanbul. TARIM BILIMLERI DERGISI 2000, 6 (4), 95-100 Mis ırda Verim ve Baz ı Verim Öğelerinin Adaptasyonu ve Stabilite Analizi H. Yavuz EMEKLIER 1 Melehat AVCI BIRSIN TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 96 ancak yüksek verim ve stabilitenin birlikte olmas ı durumunda amaca ula şılabileceğini belirtmi şlerdir. ancak yüksek verim ve stabilitenin birlikte olmas ı durumunda amaca ula şılabileceğini belirtmi şlerdir. Materyal olarak Mirco, Franca, Ant-90, P-3751, P- 3394, C-6127 ve Executive hibrid m ı s ır çeşitleri kullan ı lm ışt ı r. Heinrich ve ark. (1985). alt ı melez sorgum çe şidinde verim ve verim ö ğeleriyle üç farkl ı gelişme dönemi süresince s ıcakl ık ve yağış rejimleri aras ındaki ilişkiyi regresyon katsay ıs ı n ı kullanarak belirlemi şler ve b de ğeri 1'den küçük olan üç çe şidi stabil, b değeri 1'den olan diğer üç çe şidin ise stabil olmad ığı n ı aç ıklam ışlard ır. Ekim, 70 x 25 cm s ıra aras ı- s ıra üzeri aç ı kl ığı nda olmak üzere 5,7 bitki/ m 2 ekim s ı kl ığında, 5-6 cm derinlikte 5 m'lik bloklara 4'er s ıra yap ı lm ışt ı r. Ara şt ırma y ı llara göre tekrarlanan tesadüf bloklar ı nda 4 tekrarlamal ı olarak yürütülmüştür. Ilk dört hafta içinde parseilerde seyreltme işlemi uygulanm ışt ı r. Ekim s ıras ında diamonyum fosfat (DAP) ve I. ve Il. çapadan sonra 15 kg/da % 33'lük amonyum nitrat gübresi uygulanm ışt ır. Sulama suyu ihtiyac ı " Blaney ve Cridle" metoduna göre saptanarak y ı lda iki kez yap ı lm ışt ı r. Atatürk Orman Çiftili ğinde kar ı k sulama, Ankara Üniversitesi Ziraat Fakültesi Kenan Evren Ara şt ı rma Uygulama Çiftli ğinde ise ya ğmurlama sulama yöntemiyle her sulamada 90 mm'lik su verilmi ştir. Kang ve Gorman (1989), oniki farkl ı lokasyonda yeti ştirilen onyedi melez m ıs ı r çeşidinde genotip x çevre etkileşimini önemli bulmuşlar, stabiliteyi belirlemek için stabilite varyans ın ı kullanm ışlar ve buna göre yedi melez m ı s ı r çeşidin stabil olmad ığı n ı aç ı klam ışlardir. Zencirci ve ark. (1990), Orta Anadolu Bölgesi'ne uygun buğday çeşidinin belirlenmesi için, farkl ı arşt ı rıc ılar taraf ından geli ştirilen de ğ işik stabilite istatistiklerini karşı laşt ırm ışlar; stabilite istatistiklerinin birkaç ı birarada kullan ıld ığı nda ba şar ı n ın daha da artaca ğın ı , belli bir yöre için çe şit önermede, stabilite yan ında çeşit seçiminde; agronomik, morfolojik , patolojik ve teknolojik özelliklerin de göz önünde bulundurulmas ı gerekti ğini belirtmişlerdir. Bitki biyolojik verimi: Her parselde be ş bitkinin ayr ı ayr ı toplam a ğı rl ı klar ı hasat döneminde tart ı larak, Bitki biyolojik verimi: Her parselde be ş bitkinin ayr ı ayr ı toplam a ğı rl ı klar ı hasat döneminde tart ı larak, Bin tane a ğı rl ığı : Hasat sonunda elde edilen üründen 4 x 100 adet tanenin a ğ irliğin ın ortalamas ın ı 10 ile çarparak, Özgen (1994), k ış l ık arpa çeşit ve hatlar ı n ı verim ve verim öğelerinin adaptasyon ve stabilite özelliklerini karşı laşt ırm ış; çeşitlerin çevreye tepkisinin ölçüsü olarak b değerini, stabilite parametreleri olarak S 2d ve r2 de ğerlerini kullanm ış, çeşitlerde stabilite ve adaptasyon yeteneğinin artmas ıyla birlikte verimin de artt ığı n ı belirlemiştir. Birim alan tane verimi: Bir metrekaredeki bitkilerin tane a ğı rl ıklar ı tart ılarak bulunmuştur. Elde edilen verilere E.Ü. Ziraat Fakültesi'nde geliştirilen TAR1ST paket program ı kullan ı larak y ı llar ve yerler üzerinden birle ştirilmiş varyans analizi yap ılm ış , m ıs ır çe şitlerinin çevre ile uyumunu belirleyen stabilite parametreleri ( X, b, a, S 2 d) saptanm ışt ı r. Bu araşt ı rman ın amac ı , Orta Anadolu Bölgesi'nde yeti ştirilen baz ı m ıs ı r çeşitlerinde verim ve verim ö ğelerinin adaptasyon ve stabilite özelliklerini belirlemektir. Giriş Sprague ve Federer (1951), farkl ı çevre koşullar ında yetiştirilen m ıs ır bitkilerine ait verilen analiz etmi şler, çift Heinrich ve ark, (1983), sorgumda stabil olmayan çeşitlerin olanlara oranla daha yüksek verimli oldu ğunu, Yaprak büyüme oran ı (YABO): [ YS: yaprak say ıs ı , KYAU: koçan yapra ğı aya uzunluğ u KYAG: koçan yapra ğı aya genişliği, DBKGG: döllenme ba şlan- g ı ç ı na kadar geçen süre] eşitliği yard ı m ıyla cm 2 / gün olarak hesaplamayla ( Nevado ve Cross 1990, Samanc ı 1991), Bachireddy ve ark. (1992), otuz farkl ı şeker m ıs ı r melezinde genotip x çevre ili şkisini karşılaşt ırmak için yaln ız verim ortalamas ı , Kang taraf ından geliştirilen stabilite varyans istatisti ğinin dikkate al ı nd ığı s ı ralama (KRS) yöntemi ve stabilite varyans istatisti ğinin dikkate al ı nd ığı (KMR) yöntemleri kullanm ışlar; KRS yönteminin genotiplerin stabilitesini belirlemek için kullan ılabileceğ ini belirtmişlerdir. Hasatta tane nemi: Her parselden rastgele al ınan üç koçan ın taneleri harman edilerek, tane nemi f ı r ı n kuru yöntemiyle, TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 Çiçeklenme zaman ında ortadaki iki s ırada be ş bitki rastgele etiketlenmi ş, hasat zaman ında bu bitkiler toprak yüzeyinden orakla biçilerek al ı nm ış ve aşağıdaki özelliklere ili şkin veriler bu materyal üzerinde çal ışı larak saptanm ışt ı r. Bitki boyu: Tepe püskülü ile toprak yüzeyi aras ı nda kalan dikey uzunluk süt olum döneminde cm olarak ölçülerek (Genter ve Camper 1973, Anonim 1980, Emeklier 1985, Alt ı nbaş 1988, Kang ve Pham (1991), farkl ı uluslararas ı m ıs ır verim denemesinde Hühn'ün S, 3 ve S6 (her çevrede genotiplerin verim s ıralamas ı) istatisti ği ile Lin ve Binn'in P, (bütün lokasyon ortalamalar ında maksimum tepki ve kareler ortalamas ı uzakl ığı) istatistiği aras ındaki ilişkiyi ara şt ırd ı klar ı çal ışmalar ı nda, S, 3 ve S, 6 istatistiklerinin verim stabilitesi için yap ılacak seçmelerde kullan ılabileceğini aç ı klam ışlard ı r. Yaprak büyüme oran ı (YABO): YABO = YS x KYAU x KYAG x 0.5 / DBKGS (gün) [ YS: yaprak say ıs ı , KYAU: koçan yapra ğı aya uzunluğ u KYAG: koçan yapra ğı aya genişliği, DBKGG: döllenme ba şlan- g ı ç ı na kadar geçen süre] eşitliği yard ı m ıyla cm 2 / gün olarak hesaplamayla ( Nevado ve Cross 1990, Samanc ı 1991), Yaprak büyüme oran ı (YABO): Bitki boyu M ıs ır çeşitlerinde bitki boyu bak ım ından ele al ınan stabilite parametreleri Çizelge 3'te verilmi ştir. Çizeige incelendiğinde; Mirco ve Executive çe şitlerinin b ve a değeri aç ıs ından ideal tipe yak ın olduğu görülmektedir (Şekil 2). P-3394 çe şidi ise genel ortalaman ın üstündeki bitki boyu ve yüksek b değeri ile iyi çevre ko şullarına iyi uyum gösterebilecektir ( Şekil 2). Ant-90 ve C-6127 çeşitleri ise bitki boyu bak ım ından yüksek b de ğeriyle genel ortalaman ın alt ında kalm ışt ır. Franca çe şidi de genel ortalaman ın alt ındaki bitki boyu, küçük b de ğeri ve yüksek a ve S 2d değerleriyle bölgeye uyum gösterememi ştir. Materyal ve Yöntem M ıs ı rda verim ve baz ı verim öğelerine ili şkin y ı llar ve yerler üzerinden birle ştirilmi ş varyans analizi sonuçlar ı Çizelge 1'de verilmi şitir. Çizelge incelendi ğinde; verim ve diğer özellikler bak ım ından çe şit x çevre etkileşiminin istatistiksel olarak önemli oldu ğu görülmektedir. Araşt ırma, 1994 ve 1995 y ı llar ı nda Atatürk Orman Çiftliği ve Ankara Üniversitesi Ziraat Fakültesi Kenan Evren Ara şt ırma Uygulama Çiftli ği olmak üzere iki lokasyonda yürütülmü ştür. EMEKLIER, H. Y. ve M. AVCI BIRSIN, "M ıs ı rda verim ve baz ı verim öğelerinin adaptasyonu ve stabilite analizi" 97 Çizelge 1. M ıs ı rda verim ve baz ı verim öğelerine ilişkin varyans analizi sonuçlar ı (kareler ortalamas ı ) Çizelge 1. M ıs ı rda verim ve baz ı verim öğelerine ilişkin varyans analizi sonuçlar ı (kareler ortalamas ı ) V.K. S.D. Birim alan tane verim Bitki boyu YABO Hasatta tane nemi Bitki biyolojik verimi Bin tane a ğı rl ığı Genel 111 86758.75 389.34 198.73 5.77 45035.19 2727.01 Çevre 3 2165245 85 6553.12 * 4018.16 97.19 637539.64 52114.93 Hatal 12 51119.36 372.02 132.78 11.19 29116.50 221.58 , Genotip 6 58438.97* 1066.50 683.35 18.12 272443.57 9615.6 G _____çx 18 33248.32* 176.35 85.93 2.05 18114.44** 3096.62 Hatat 72 21832.61 132.22 38.40 0.96 10780.11 420.29 * % 5, ** % 1 düzeyinde önemli * % 5, ** % 1 düzeyinde önemli Öte yandan C-6127 ve Executive çe şitleri küçük b de ğeri, ortalaman ın alt ındaki verimleri ve büyük S 2d ve a değerleri nedeniyle bölgeye uyum sa ğlayamad ıklar ı söylenebilir. Öte yandan C-6127 ve Executive çe şitleri küçük b de ğeri, ortalaman ın alt ındaki verimleri ve büyük S 2d ve a değerleri nedeniyle bölgeye uyum sa ğlayamad ıklar ı söylenebilir. Ara şt ırmada ele al ı nan stabilite parametrelerine göre verimi ortalama yerimden yüksek, regresyon katsay ıs ı (b) 1'e yak ın, a katsay ıs ı küçük ve regresyondan sapma kareler ortalamas ı (S2 d) O'a yak ın olan çeşitler istenen çeşitler olarak değerlendirilmi şlerdir ( Eberhart ve Russelle 1966, Zencirci ve ark. 1990). Bitki alan tane verimi M ıs ır çeşitlerinde verim bak ım ından ele al ınan stabilite parametrelerine ili şkin değerler Çizelge 2'de verilmiştir. Çizelge incelendi ğinde; P-3394, P-3751 ve Franca çe şitlerinde verimin ortalaman ın üstünde, b değerinin ise l'e yak ın olduğu görülmektedir. b de ğerinin l'e yak ın olmas ı çeşitlerin çevre ko şuflar ındaki değ işime iyi uyum sağlad ığı n ı göstermektedir (Finlay ve Wilkinson 1963, Eberhat ve Russell 1966) ( Şekil 1). Öte yandan küçük S2 d ve a değerleriyle Franca ve P-3751 çe şitlerinin bölgenin her türlü ko şullar ında verimli bir şekilde yetiştirilebilece ği anlaşılmaktad ır. Mirco ve Ant-90 çeşitlerinde verim ortalaman ın alt ında olmas ına karşın b değeri 1, a ve S2 d değerleri küçük ç ıkm ışt ır. b değeri bak ı m ından güvenlik s ı n ırlar ı içinde kalan bu çe şitler ( Şekil 1) tüm bölgeler için önerilebilir (Finlay ve Wilkinson 1963, Henrich ve ark. 1985). Çizelge 3. Farkl ı çevrelerde yetiştirilen m ıs ı r çeşitlerinde bitki boyuna ilişkin adaptasyon ve stabilite parametreleri Çeşitler Ortalama b a S2d P-3751 260.7 0.97 -4.68 31.3 P-3394 281.1 1.18 -41.06 141.0 C-6127 267.8 1.05 -20.40 8.6 Franca 271.4 0.58 111.62 50.6 Mirco 276.9 1.02 -0.89 33.4 Ant-90 265.8 1.20 -63.07 10.5 Executive 282.6 0.97 18.49 32.2 Genel ort. 272.3 Çizelge 2. Farkl ı çevrelerde yeti ştirilen m ıs ı r çeşitlerinde tane verimine ilişkin adaptasyon ve stabilite parametreleri Çeşitler Ortalama b a S`d P-3751 1152.82 1.19 -221.29 2178.1 P-3394 1266.27 0.91 212.28 38893.1 C-6127 1092.83 0.84 121.97 9031.2 Franca 1184.10 1.13 -122.49 1040.2 Mirco 1145.27 1.01 -23.80 852.4 Ant-90 1119.83 1.00 -32.27 6456.4 Executive 1094.96 0.89 65.60 4886.0 Genel ort. 1150.86 r ♦P-3751 E P-3394 (5 C-6127 X Franca X Mirco •Ant-90 +Executive 1,4 1,2 - 1 e • IOi I X 0,8 - 0,6 - X 0,4 - 0,2 - 0 255 260 265 270 275 280 285 Bitki boy u(cm) Şekil 2. M ıs ı r çeşitlerinin bitki boyu ortalamas ı ve regresyon katsay ıs ı na göre adaptasyon durumu Reg resyon katsay ıs ı r ♦P-3751 E P-3394 (5 C-6127 X Franca X Mirco •Ant-90 +Executive ♦P-3751 I P-3394 PC-6127 XFranca Mirco •Ant-90 4-Execuliv e 1,4 1,2 x 1 • X 0,8 I 0,6 0,4 0,2 11 O O 1150 1200 1250 1300 Verim (kg/da) Şekil 1. Hasatta tane nemi M ı s ır çeşitlerinde hasatta tane nemi bak ım ından ele al ı nan stabilite parametreleri Çizelge 5'te verilmi ştir. Çizelge incelendiğinde, P-3751, P-3394 ve C-6127 çeşitlerinin genel ortalaman ın alt ında nem değerine sahip olduğu görülmektedir. M ı s ır çeşit ıslah ı nda adaptasyon denemelerinde, çeşitlerin hasat döneminde tane nemlerinin düşük oranda olmas ı istenilen bir özelliktir ( Emeklier 1997). Bu özellik bak ı m ı ndan P-3751 ve C- 6127 çeşitleri dü şük nem oran ı , l'e yak ın b değeri ile küçük a ve S2d değerleriyle uyumludur ( Şekil 4). Franca ve Executive çe şitlerinin genel ortalaman ın üstündeki tane nemi, 1'in alt ındaki b değeri ve büyük a ve S 2d değerleriyle bölgeye uyum gösterememi ştir. M ıs ır çeşitlerinde yaprak büyüme oran ı bak ı m ı ndan ele al ınan stabilite parametreleri Çizelge 4'te verilmi ştir. Çizelge incelendiğinde P-3394, Mirco çe şitlerinde yaprak büyüme oran ı n ı n genel ortalaman ın üstünde ve b değerinin 1 'e yak ı n olduğu görülmektedir. Mirco çe şidi l'e yak ın b değeri , küçük a ve s 2d değerleriyle tüm çevrelere önerilebilecektir ( Şekil 3). Bu özellik bak ı m ından C-6127, Ant-90 ve Executive çe şitleri 1'e yak ın b değeri göstermi ş , ancak genel ortalaman ı n alt ındaki yaprak büyüme oran ı ve büyük a ve S 2d değerleriyle bölgeye uyum sağlayamam ışt ı r. Çizelge 5. Farkl ı çevrelerde yetiştirilen m ıs ı r çeşitlerinin hasatta tane nemine ilişkin adaptasyon ve stabilite parametreleri Çizelge 4. Farkl ı çevrelerde yeti ştirilen m ıs ı r çeşitlerinde yaprak büyüme oran ı na ili şkin adaptasyon ve stabilite parametreleri Çizelge 5. Farkl ı çevrelerde yetiştirilen m ıs ı r çeşitlerinin hasatta tane nemine ilişkin adaptasyon ve stabilite parametreleri Çeşitler Ortalama b a Szd . P-3751 8.39 1.14 -3.27 0.3 P-3394 10.07 0.77 2.14 0.7 C-6127 9.71 1.31 -3.69 0.4 Franca 11.79 0.76 4.03 0.5 Mirco 10.85 1.19 -1.39 0.3 Ant-90 10.39 1.13 -1.17 0.1 Executive 10.28 0.67 3.37 0.3 Genel ort. 10.21 Çeşitler Ortalama b a S`d P-3751 74.70 0.79 6.68 25.2 P-3394 92.73 1.24 -14.12 35.8 C-6127 82.38 1.08 -11.02 16.3 Franca 90.29 0.58 39.81 5.9 Mirco 93.01 0.98 8.14 0.5 Ant-90 84.87 1.17 -16.27 29.3 Executive 84.81 1.13 -13.22 9.4 Genel ort. TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 98 Bitki alan tane verimi M ıs ır çeşitlerinin verim ortalamas ı ve regresyon katsay ıs ına göre adaptasyon durumu Regresyon katsay ısı 0 1050 ♦P-3751 I P-3394 PC-6127 XFranca Mirco •Ant-90 4-Execuliv e 1,4 1,2 x 1 • X 0,8 I 0,6 0,4 0,2 11 O O 1150 1200 1250 1300 Verim (kg/da) Şekil 1. M ıs ır çeşitlerinin verim ortalamas ı ve regresyon katsay ıs ına göre adaptasyon durumu Regresyon katsay ısı 0 1050 Bin tane ağı rl ığı M ıs ır çeşitlerinde koçan hasad ından sonra tarlada kalan bitkilerin has ıl m ıs ır ya da silaj m ıs ır olarak değerlendirilmesi aç ıs ından biyolojik verimin yüksek olmas ı istenmektedir (Emeklier 1997).M ıs ır çeşitleri sapl ı ağı rl ık bak ım ından tüm parametreler ile değerlendirildi ğinde, P-3394 ve Mirco çe şitleri genel ortalaman ın üstündeki verimleri, l'e yak ın b değerleri ve düşük a ve S2d değerleriyle bölgeye uyum göstermi ştir (Çizelge 6, Şekil 5). Franca çe şidi ise genel ortalaman ın üstündeki verimi ve dü şük b değeriyle bölgenin olumsuz koşullar ına iyi uyum sağlayabilmiştir (Şekil 5). Bu özellik bak ım ından P-3751, C-6127, Ant-90 ve Executive çe şitleri genel ortalaman ın alt ında kalm ış ve l'e yak ın b değeri , yüksek a ve S2d değerleriyle bölgeye zay ıf adaptasyon göstermi ştir. M ıs ır çeşitlerinde bin tane a ğı rl ığı bak ım ından ele al ınan stabilite parametreleri Çizelge 7:çle verilmi ştir. Çizelge incelendi ğinde, C-6127,,Mirco ve Ant-90 çe şitleri genel ortalaman ın üstündeki bin tane ağı rl ığı , 1'e yak ı n b Q ğeri, küçük a ve ,,S2d değerleriyle bölgeye uyurp,. sağlam ış ve stabil bir görünüm vermi ştir ( Şekil 6). C-6127 ve Executive çeşitleri ortalamadan yüksek bin tane a ğı rl ığı ve düşük b değeri ile bölgenin olumsuz koşullar ı na iyi uyum sağlayabilecektir (Şekil.. 6). p-375 t ve Franca çeşitleri ise 1'e yak ın b değeri ile genel ortalamay ı geçememiş ve bölgenin tüm ko şullarına uygun adaptasyon gösterememi ştir. Çizelge 7.Farkl ı çevrelerde yetiştirilen m ıs ı r çeşitlerinin bin tane ağı rlığı na ilişkin adaptasyon ve stabilite parametreleri Çizelge 6. Farkl ı çevrelerde yeti ştirilen m ıs ı r çeşitlerinin bitki biyolojik verimine ilişkin adaptasyon ve stabilite parametreleri Çeşitler Ortalama b a S2d P-3751 818.87 1.03 -228.96 1812.8 P-3394 1166.80 1.31 -164.15 5718.1 C-6127 945.72 0.82 112.12 2019.0 Franca 1106.32 0.67 412.74 16089.3 Mirco 1147.32 1.03 98.81 4307.1 Ant-90 941.20 1.09 -171.09 2818.5 Executive 961.75 1.01 -68.47 741.6 Gene! ort. 1012.56 Çizelge 6. Hasatta tane nemi 86.11 1,4 _ 1 ,2 ' (T) 1 0,8 5; 0,6 <ı) 6, 0,4 0,2 0 90 100 50 60 70 80 Yaprak büyüme oran ı Şekil 3. M ı s ı r çeşitlerinin yaprak büyüme oran ı ortalamas ı ve regresyon katsay ıs ına göre adaptasyon durumu 5 7 9 11 13 I e x ♦P-3751 I P-3394 AC-6127 XFranca XMirco •Ant-90 +Executive •P-3751 I P-3394 AC-6127 X Franca X Mirco •Ant-90 +Executive 1,4 ,_ 1 2 nı 1 tıı 2 0,8 c 5), 0,6 '6) 0,4 aı 0,2 0 Hasatta tane nemi (%) Şekil 4. M ı s ı r çeşitlerinin hasatta tane ne ortalamas ı ve regresyon katsay ı s ı na göre adaptasyon durumu 1,4 _ 1 ,2 ' (T) 1 0,8 5; 0,6 <ı) 6, 0,4 0,2 0 90 100 50 60 70 80 Yaprak büyüme oran ı Şekil 3. M ı s ı r çeşitlerinin yaprak büyüme oran ı ortalamas ı ve regresyon katsay ıs ına göre adaptasyon durumu I e x ♦P-3751 I P-3394 AC-6127 XFranca XMirco •Ant-90 +Executive 1,4 _ 1 ,2 ' (T) 1 0,8 5; 0,6 <ı) 6, 0,4 0,2 0 90 100 50 60 70 80 Yaprak büyüme oran ı Şekil 3. M ı s ı r çeşitlerinin yaprak büyüme oran ı ortalamas ı ve regresyon katsay ıs ına göre adaptasyon durumu I e x ♦P-3751 I P-3394 AC-6127 XFranca XMirco •Ant-90 +Executive 5 7 9 11 13 •P-3751 I P-3394 AC-6127 X Franca X Mirco •Ant-90 +Executive 1,4 ,_ 1 2 nı 1 tıı 2 0,8 c 5), 0,6 '6) 0,4 aı 0,2 0 Hasatta tane nemi (%) Şekil 4. M ı s ı r çeşitlerinin hasatta tane ne ortalamas ı ve regresyon katsay ı s ı na göre adaptasyon durumu 99 EMEKLiER, H. Y. ve M. AVCI BIRSIN, "M ıs ırda verim ve baz ı verim öğelerinin adaptasyonu ve stabilite analizi" Kaynaklar Kang, M. S. ve D. P. Gorman, 1989. Genotype x environment interaction in maize. Agron. J. 81:662-664. Alt ı nbaş, M. 1988. M ı s ı rda Erkenciliğin Kal ıt ım ı ve Bunun Baz ı Tar ı msal Özellikler ile Ili şkileri Üzerinde Ara şt ı rmalar. Ege Üniversitesi Fen Bilimleri enstitüsü Tarla Bitkileri ABD, Doktora Tezi, Izmir, 174 s. Kang, M. S. ve H. N. Pham, 1991. Simultaneous selection for high yielding and stable crop genotypes. Agron. J. 83: 161- 165. Anonim, 1980. International Union For The Protection of New Varieties of Plants (UPOV). Guidelines for the conduct of tests for distinctness, homogeneity and stability. Maize TG/2/4. Nevada, M. E. ve H.Z. Cross, 1990. Diallel analysis of relative growth rates in maize synthetics. Crop Science, 30: 549- 552. Özgen, M. 1994. Orta Anadolu ko şullar ı nda k ış l ı k arpan ı n verim ve verim ö ğelerinde adaptasyon ve stabilite analizi. Tr. 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Ziraat Fakültesi Diploma Sonras ı Yüksek Okulu Tarla Bitkileri Bölümü, Doktora Tezi. Ankara 129 s. Zencirci, N., V. Eser ve I. Baran, 1990. Baz ı stabilite istatistiklerinin karşı laşt ı r ı lrnas ı üzerine bir yakla şım. Tarla Bitkileri Mer. Aras. Enst. Yay no: 1990/2, Ankara. Emeklier, H. Y. 1997. Erkenci Hibrid M ıs ı r Çeşitlerinin Verim ve Fenotipik Özellikleri Üzerine Ara şt ı rmalar. Ankara Üniv. Zir. Fak. Yay. TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 100 Sonuç Finlay, K. W. ve G. N. Wilkinson, 1963. The analysis of adap- tation in a plar ıt breeding programme, Aust. J. Agric. Res., 14: 742-754. Ankara koşullar ında 1994 ve 1995 y ıllarında iki farkl ı lokasyonda yetiştirilen yedi hibrid m ıs ır çeşidinde verim ve baz ı verim öğelerinin adaptasyon ve stabilite özelliklerinin belirlendi ği bu çal ışmada; verim yönünden P-3751, P- 3394 ve Franca çe şitlerinin diğer çeşitlere oranla tüm çevrelerde daha uyumlu ve stabil oldu ğu görülmüştür. Gelecekte yap ı lacak adaptasyon çal ışmalar ın ın daha fazla çe şit ve değ işik stabilite parametreleri kullan ılarak yürütülmesi bölgeye uygun çe şit seçiminin isabetli yap ılmas ına olanak sağlayacakt ır. Ayr ıca yüksek verim ve stabilitenin birlikte olmas ı (Henrich ve ark. 1983) ve çe şit seçiminin de agronomik, morfolojik, patolojik ve teknolojik özelliklerini de dikkate al ı nmas ı n ı n (Zencirci ve ark.1990) yararl ı olaca ğı söylenebilir. Francis, T. R. ve L. W. Kannenberg, 1978. Yield stability studies in short-season maize. I. A descriptive method for grouping genotypes. Can. J. Plant Sci. 58: 1029-1034. Genter, C. F. ve Jr. H. M. Camper, 1973. Component plant part development in maize as affected by hybrids and population density. Agronomy Journal, 65: 669-671. Heinrich, G. M., C. A. Francis ve J. D. Eastin, 1983. Stability of grain sorghum yield components across diverse environments. Crop Science, 23: 209-212. Heinrich, G. M., C. A. Francis ve J. D. Eastin ve M. Saeed, 1985. Mechanisms of yield stability in sorghum. Crop science, 25: 1109-1112. Bin tane ağı rl ığı Farkl ı çevrelerde yeti ştirilen m ıs ı r çeşitlerinin bitki biyolojik verimine ilişkin adaptasyon ve stabilite parametreleri Çeşitler Ortalama b a S2d P-3751 282.50 1.29 -132.19 934.0 P-3394 321.25 0.69 98.00 931.0 C-6127 322.12 1.08 -24.24 522.9 Franca 294.25 1.05 -42.57 1762.3 Mirco 351.81 1.02 22.90 914.8 Ant-90 321.62 1.11 -35.22 453.6 Executive 343.12 0.71 113.33 662.1 Genel ort. 319.52 • P-3751 I P-3394 AC-6127 XFranca X Mirco +Executive 1,4 I 1,2 - .1 - • 111+ 0,8 - 0,6 - x 0,4 - 0,2 - O 500 700 900 1100 1300 Sapl ı ağı rl ık (g/bitki) Şekil 5. M ıs ı r çeşitlerinin sapl ı ağı rl ı k ortalamas ı regresyon katsay ıs ı na göre adaptasyon durumu Regresyon katsay ıs ı • P-3751 I P-3394 AC-6127 XFranca XMirco •Ant-90 +Executive 1,4 • 1,2 1 0,8 0,6 0,4 0,2 0 200 250 300 350 400 Bin tane ağırl ığı(g) Şekil 6. M ıs ı r çeşitlerinin bin tane a ğı rl ığı ortalama ve regresyon katsay ıs ı na göre adaptasyon durumu Regresyon katsay ıs ı • P-3751 I P-3394 AC-6127 XFranca X Mirco +Executive 1,4 I 1,2 - .1 - • 111+ 0,8 - 0,6 - x 0,4 - 0,2 - O 500 700 900 1100 1300 Sapl ı ağı rl ık (g/bitki) Şekil 5. M ıs ı r çeşitlerinin sapl ı ağı rl ı k ortalamas ı regresyon katsay ıs ı na göre adaptasyon durumu Regresyon katsay ıs ı • P-3751 I P-3394 AC-6127 XFranca XMirco •Ant-90 +Executive 1,4 • 1,2 1 0,8 0,6 0,4 0,2 0 200 250 300 350 400 Bin tane ağırl ığı(g) Şekil 6. M ıs ı r çeşitlerinin bin tane a ğı rl ığı ortalama ve regresyon katsay ıs ı na göre adaptasyon durumu Regresyon katsay ıs ı Şekil 6. M ıs ı r çeşitlerinin bin tane a ğı rl ığı ortalama ve regresyon katsay ıs ı na göre adaptasyon durumu TARIM BILIMLERI DERGISI 2000, Cilt 6, Say ı 4 Kaynaklar No:1493, Bilimsel Ara şt ı rmalar ve Incelemeler: 817, Ankara, 68 s.	50,138
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https://openalex.org/W3129292335_2	Spanish-Science-Pile	Open Science	Various open science	2,021	La educación moral en las primeras edades a través del teatro español del siglo XVIII y primera mitad del XIX	None	Spanish	Spoken	1,392	2,530	Ediciones Universidad de Salamanca Hist. educ., 39, 2020, pp. 231-248  la educación moral en las primeras edades a través del teatro español del siglo xviii y primera mitad del xix federico briante benítez en sus modales la honestidad con la elegancia; porque ni gusta del desaliño, ni de la demasía y afectación en el adorno»53. En esta línea caminan las opiniones de Josefa Amar y Borbón54, Jovellanos (en su primera sátira «A Arnesto»)55 o el anónimo autor56 de la citada carta publicada por Clavijo y Fajardo en El Pensador; todos ellos critican con dureza la afición de los jóvenes por la frivolidad y los lujos excesivos. Esta llamada a educar en la moderación es lo que subyace al acto II de La bella pastora, de Bruno Solo de Zaldívar; en esta comedia, el dramaturgo aprovecha la conducta ejemplarizante de Rosaura para hacer un encomio de las madres que educan a sus hijas en la contención y en no ser «loca[s], presumida[s] y vana[s]»57. Merece la pena reproducir parte del fragmento: Partenio Benditas sean las madres, que a sus hijas no defraudan de un patrimonio tan grande, como la prudente, sabia educación, que en el mundo no hay tesoro que más valga. Marcela Es verdad. Rosaura barriendo. Partenio ¡Oh, qué ejemplar tan útil, si le tomaran muchas madres con sus hijas, en lugar de las erradas máximas de dirigirlas al uso, al fausto, a la gala y otros lucimientos, que  Morales, José Isidoro: «Comentario de D. José Isidoro y Morales al Excmo. Sr. D. Joseph Mazarredo sobre la enseñanza de su hija», Escritos pedagógicos de la Ilustración (vol. I), Madrid, Ministerio de Educación y Ciencia, 1988, p. 285.  «En efecto, es cosa ridícula que haya madres que contenten los deseos de sus hijas en cuantas diversiones apetecen, en la variedad de vestidos y de adornos y que no piensen en destinar una tercera parte de lo que en esto consumen en procurarles la adquisición de la verdadera sabiduría» [Amar y Borbón, Josefa: Discurso sobre la educación física..., op. cit., p. 310].  «Y tú, mísera España, tú la esperas / sobre la playa, y con afán recoges / la pestilente carga y la repartes / alegre entre tus hijos. Viles plumas, / gasas y cintas, flores y penachos, / te trae en cambio de la sangre tuya, / de tu sangre ¡oh baldón!, y acaso, acaso / de tu virtud y honestidad. Repara / cuál la liviana juventud los busca. / Mira cuál va con ellos engreída / la imprudente doncella; su cabeza, / cual nave real en triunfo empavesada, / vana presenta del favonio al soplo / la mies de plumas y de agrones, y anda / loca, buscando en la lisonja el premio / de su indiscreto afán [...]» [Jovellanos, Gaspar Melchor de: Poesías de Gaspar Melchor de Jovellanos, Oviedo, Instituto de Estudios Asturianos, edición de José Caso González, 1961, p. 239].  «Es muy notable y digno de compasión que un padre que no perdona gasto alguno para procurar a sus hijos establecimientos brillantes, y que a costa de gastos excesivos les mantiene magníficos vestidos y equipajes y multitud de criados inútiles, sólo conozca la economía cuando se trata de buscar y atraer una o más personas que cultiven el espíritu de los mismos hijos [...] aquellos gastos se sacrifican al orgullo y la vanidad, sin tener parte en ellos el sincero deseo de hacer bien a sus hijos» [Labrador, Carmen y Pablos, Juan Carlos de: La educación en los papeles..., op. cit., p. 232].  Solo de Zaldívar, Bruno: La bella pastora y ciudadana en el monte y discreto labrador, en la Biblioteca Histórica Municipal de Madrid (Ms. Tea 1-12-10, B), s. a., f. 18v. Ediciones Universidad de Salamanca Hist. educ., 39, 2020, pp. 231-248 la educación moral en las primeras edades a través del teatro español del siglo xviii y primera mitad del xix federico briante benítez  cuestan mucho y valen nada! Rosaura Dice usted bien y yo digo Barriendo. que esas madres son madrastras58. En el polo opuesto a Rosaura tenemos a Ceferina, La niña desdeñosa de Luciano Francisco Comella59. Ceferina representa el prototipo de la adolescente difícil, soberbia, caprichosa y amante de las modas; como suele ocurrir en otros casos vistos en estas páginas, la mala educación de la chica se debe, en gran parte, a la permisividad y escasa atención que le presta su padre Facundo. Pero quizá el ejemplo más notable de hasta qué punto de ruina moral y económica puede llegar una familia a causa del apego excesivo de sus miembros a la moda nos lo ofrece la comedia de María Rosa Gálvez anteriormente mencionada; en este caso, la oportuna intervención de doña Guiomar, personaje que representa «las luces de la razón entre tanta locura»60, evita que la pésima situación familiar vaya a mayores. 3. Conclusiones Los adelantos científico-médicos, la concienciación de las autoridades y de la ciudadanía en general acerca de la importancia del cuidado de la higiene y la consolidación del sentimiento moderno de familia, entre otros factores, despertaron en el siglo xviii un interés inusitado por las primeras edades de la persona y por todo lo relacionado con ellas, especialmente el tema educativo. A esto se suma que nos encontramos en el siglo de la Ilustración, un movimiento ideológico que confía en el poder edificante y regenerador de la educación para con toda la sociedad; más aún cuando de lo que hablamos es de la educación de los más jóvenes, a los que se les considera como «la esperanza y el nervio de la patria, [...] el plantel de los que algún día han de llegar a ser hombres y mantener el estado», en palabras de Jaime Bonells61. En consecuencia, de la educación que aquellos reciban en sus respectivos hogares dependerá el bien de todo el estado; como dice el también ilustrado Hervás y Panduro, «los hijos bien criados hacen felices las familias y la felicidad de estas es la del estado»62. La educación física e intelectual forma a los jóvenes como «ciudadanos útiles», pero únicamente la educación moral, de la que nos hemos ocupado en este estudio, los hará «ciudadanos útiles y buenos»63, como apostilla Jovellanos en su citada Memoria sobre educación pública; y esto gracias a la práctica de las virtudes, que, según el ideario del Siglo de las Luces, constituía la principal vía para Solo de Zaldívar, Bruno: La bella pastora y ciudadana..., op. cit., ff. 18v.-19r. Comella, Luciano Francisco: La niña desdeñosa, en la Biblioteca Histórica Municipal de Madrid (Ms. Tea 1-133-6, A), s. a.  Trueba, Virginia: El claroscuro de las Luces. Escritoras de la Ilustración española, Barcelona, Ediciones de Intervención Cultural, 2005, p.103.  Bonells, Jaime: Perjuicios que acarrean..., op. cit., p. 318.  Hervás y Panduro, Lorenzo: Historia de la vida..., op. cit., pp. 337-338.  Jovellanos, Gaspar Melchor de: «Memoria sobre educación pública», op. cit., p. 251.   Ediciones Universidad de Salamanca Hist. educ., 39, 2020, pp. 231-248 la educación moral en las primeras edades a través del teatro español del siglo xviii y primera mitad del xix federico briante benítez  alcanzar la felicidad y la fuente esencial de la nobleza humana. A este último asunto hacen referencia muchas de las obras dramáticas aquí estudiadas; así, Duling, en la comedia Cual el padre, así es el hijo, hace saber a Carlos que «la mayor nobleza / es la virtud»64; en La señorita malcriada, doña Clara apunta que «no se trata / de linajes. La conducta / es la que humilla o exalta»65; y en la comedia Cual más obligación es la de padre o la de juez, Pablo afirma que «[...] el villano se hace noble / en lo noble de sus tratos»66. A través de estos ejemplos, que vendrían a sumarse a los que se han ido desgranando a lo largo de estas líneas, es posible comprobar cómo el teatro no se mantuvo ajeno a las corrientes pedagógicas del siglo, antes bien, participó en ellas desempeñando un papel muy activo en la siempre difícil tarea de educar a los más jóvenes.    Enciso Castrillón, Félix: Cual el padre..., op. cit., f. 12v. Iriarte, Tomás de: «La señorita malcriada», op. cit., p. 139. Valladares de Sotomayor, Antonio: Cual más obligación..., op. cit., f. 9r. Ediciones Universidad de Salamanca Hist. educ., 39, 2020, pp. 231-248.	39,393
W1582611568.txt_1	German-Science-Pile	Open Science	Various open science	null	None	None	Unknown	Unknown	5,910	13,374	ETH Library Schutz flussnaher Trinkwasserfassungen bei Flussraum-Aufweitungen in voralpinen Schotterebenen Journal Article Author(s): Hoehn, Eduard; Meylan, Benjamin Publication date: 2009-12 Permanent link: https://doi.org/10.3929/ethz-b-000083773 Rights / license: In Copyright - Non-Commercial Use Permitted Originally published in: Grundwasser 14(4), https://doi.org/10.1007/s00767-009-0111-3 This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 DOI 10.1007/s00767-009-0111-3 Fachbeitrag Schutz flussnaher Trinkwasserfassungen bei Flussraum-Aufweitungen in voralpinen Schotterebenen Eduard Hoehn · Benjamin Meylan Eingang des überarbeiteten Beitrages: 5. 5. 2009 / Online veröffentlicht: 25. 6. 2009 © Springer-Verlag 2009 Kurzfassung In den meisten alpinen und voralpinen Schotterebenen bestehen Wechselwirkungen zwischen Flüssen und Grundwasser. Viele Trinkwasserversorger nutzten dies, indem sie ihre Fassungen in Flussnähe bauten. Neuerdings hat sich im Flussbau die Einsicht durchgesetzt, Hochwasserschutzmaßnahmen mit einer Aufwertung bzw. Aufweitung des Flussraums zu koppeln. Dadurch können diese flussnahen Fassungen gefährdet werden. Meist verringert sich durch Arbeiten im Flussraum die Verweildauer des Grundwassers, und der Anteil an flussnahem Infiltrat in der Fassung erhöht sich. Hochwasserereignisse können zusätzlich zu Durchbrüchen von flussnahem Infiltrat führen, was besonders bei hoher Belastung des Flusses durch Abwasser mit einer Grundwasserverunreinigung einhergeht. Wo Flussraum-Aufweitungen unumgänglich sind, müssen Maßnahmen getroffen werden, um wichtige Trinkwasserfassungen vor Verunreinigung zu schützen. Drei Fälle mit unterschiedlichem Risiko werden diskutiert: i) geringes Risiko bei fehlenden Wechselwirkungen oder bei Exfiltration, ii) Risiko bei Infiltration, während oder nach geplanten Aufweitungen, und iii) hohes Risiko bei problembehafteten Fassungen. Bei Infiltration über eine mächtige ungesättigte Zone zwischen Fluss und Fassung ist das Risiko geringer als bei Infiltration mit direktem hydraulischem Anschluss des Flusses ans Grundwasser. Für Fassungen mit Risiko bei Flussraum-Aufweitungen wer- Eduard Hoehn () Eawag, Wasserforschungs-Institut des ETH-Bereichs, 8600 Dübendorf, Schweiz E-Mail: hoehn@eawag.ch Benjamin Meylan, BAFU, Bundesamt für Umwelt, Sektion Grundwasserschutz, 3003 Bern, Schweiz den technische Maßnahmen zur Minderung des Gefahrenbildes vorgeschlagen. Measures to protect drinking-water wells near rivers from hydraulic engineering operations in peri-alpine flood-plains Abstract In alpine and peri-alpine flood plains, rivers can lose water to the ground water (infiltrating conditions), or gain water from it (exfiltrating conditions). Since sediments within these flood plains are highly permeable, drinking water wells near infiltrating rivers are often highly efficient. To reduce flood risks, engineers couple hydraulic flood protection measures with an enhancement of lateral connectivity. Eventually, the residence time of the infiltrating ground water is reduced and the fraction of infiltrated water rises. Floods lead to the breakthrough of freshly infiltrated water. This can lead to groundwater contamination, if the river is loaded with wastewater. Where measures of hydraulic engineering are necessary, drinking-water wells must therefore be protected from contamination. We discuss three cases of contamination risk: i) low risk under exfiltrating conditions or where river and ground water interaction is weak, ii) increased risk during or after hydraulic operations under infiltration conditions; and iii) generally high risk at wells with low residence times and high fractions of infiltration water. We suggest that the risk is reduced when a thick unsaturated zone separates the river from the well (> 10 m), compared to direct water loss through saturated media. For wells at risk, we propose technical protection measures to reduce the danger. Keywords flood plain · losing river · lateral connectivity · land-use conflict · residence time · mixing · drinking-water supply 123 256 Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 Einleitung In Gebieten mit regem Wasseraustausch zwischen Fluss und Grundwasser entstehen ökologisch wertvolle Auenlandschaften (Ward & Stanford 1995). Zum Schutz vor Hochwasser sowie zur Gewinnung zusätzlicher Landflächen wurden insbesondere im letzten und vorletzten Jahrhundert viele Flüsse in Mitteleuropa stark verbaut, das heißt begradigt und eingedämmt. Diese Schutzmaßnahmen führten zu den bestehenden naturfernen Flusslandschaften mit eingeschränkter Auenvegetation. Die Hochwasserereignisse der vergangenen Jahre haben gezeigt, dass viele dieser Verbauungen den heutigen Sicherheitsanforderungen nicht mehr genügen. Dabei wurde klar, dass den Fließgewässern aus Gründen der Hochwassersicherheit wieder vermehrt Platz eingeräumt werden muss. Auch aus ökologischer Sicht sind Fließgewässer zu revitalisieren. Damit soll zumindest teilweise wieder eine naturnahe Fließwasserdynamik mit Wechselwirkungen zum Grundwasser hergestellt und die mit Dämmen vom Flusslauf getrennten Auenreste wieder an den Fluss angebunden werden. Die notwendigen wasserbaulichen Eingriffe für eine solche Fließgewässer-Revitalisierung, z. B. Aufweitung des Flussraums, Uferanrisse oder Unterlassung von Unterhaltsarbeiten, können bei flussnahen Trinkwasserfassungen zu Konflikten mit der Grundwassernutzung führen. Gemäß der EU-Wasserrahmenrichtlinie (WRRL) sind Grundwasserkörper dann weitergehend zu beschreiben, wenn Wechselwirkungen mit dem entsprechenden Fließgewässer festgestellt werden. Vor der Errichtung von Dämmen (z. B. für Hochwasserschutz) müssen die Auswirkungen dieses Eingriffs auf das Grundwasser geprüft werden. In der Schweiz regelt das Wasserbaugesetz, dass wasserbauliche Eingriffe zum Schutz vor Hochwasserschäden mit Flussraum-Aufwertungen einhergehen müssen, um wieder naturnähere Flusslandschaften zu ermöglichen (Woolsey et al. 2007). Gewässer und Ufer müssen so gestaltet werden, dass die Wechselwirkungen zwischen Fluss und Grundwasser weitgehend erhalten bleiben. Die schweizerische Auenschutzverordnung fordert solche Aufwertungsmaßnahmen auch ohne Eingriffe zugunsten des Hochwasserschutzes. Als Arbeitshypothese gehen wir davon aus, dass flussnahe Trinkwasserfassungen gegenüber einer Verunreinigung des Grundwassers gefährdet sind (Trüeb 1962, 1963, Sontheimer & Nissing 1977). Daraus folgt, dass die zuständigen Behörden den potenziellen Konflikt von FlussraumAufweitungen mit der Grundwassernutzung im flussnahen Bereich kennen und entsprechende Schutzmaßnahmen ergreifen müssen. Die praktischen Möglichkeiten zur Vermeidung bzw. Entschärfung eines solchen Konflikts wurden unseres Erachtens bisher noch zu wenig ausgeleuchtet. Ziel dieser Arbeit ist es deshalb, das Risiko einer Beeinträchtigung flussnaher Fassungen zu beurteilen und technische 123 Maßnahmen zu ihrem Schutz vorzuschlagen. Dies ist umso besser möglich, je genauer die Wechselwirkungen zwischen Grundwasser und Oberflächenwasser (physikalische und biogeochemische Vorgänge an der Grenze Flusswasser/ Grundwasser) vor und nach Flussraum-Aufweitungen bekannt sind. Grundlagen Im Folgenden werden die bisherigen Kenntnisse über die Wechselwirkungen zwischen Fließgewässern und dem Grundwasser gewürdigt und die dazu notwendigen Grundlagen dargelegt. In Schwemmebenen alpiner und voralpiner Einzugsgebiete (z. B. Münchner Schotterebene, Rheintal, Schweizerisches Mittelland) stehen Fließgewässer vielerorts im Austausch mit ungespanntem Grundwasser, mit Raten von bis zu 10 m3 m–2 d–1 (Hoehn 2002). Die Beziehungen zwischen Fließgewässer und Grundwasser sind räumlichen und zeitlichen Veränderungen unterworfen (Trüeb 1962, 1963, Brunke et al. 2003). Kiessandige Schotterablagerungen wirken als sehr gut durchlässige Grundwasserleiter (kf -Werte von im Allgemeinen 10 –4–10–2 m s–1), und die Sohlen der Fließgewässer sind im naturnahen Zustand mehrheitlich gut durchlässig. Falls der Grundwasserspiegel in Flussnähe höher liegt als der Flusswasserspiegel, besteht ein Gefälle zum Fluss, d. h. dieser wirkt als Vorfluter und das Grundwasser exfiltriert. Bei umgekehrtem Gefälle verliert der Fluss Wasser in den Untergrund und das Flusswasser infiltriert (z. B. Woessner 2000). Infiltration kann durch Pumpbetrieb in einer nahe am Ufer gelegenen Grundwasserfassung künstlich erzeugt oder verstärkt werden. Unter gewissen Bedingungen wird die Durchlässigkeit der Flusssohle durch die Bildung einer Kolmationsschicht reduziert, im Extremfall bis hin zur vollständigen Kolmation (z. B. Schälchli 1995). Kolmation vermindert bzw. unterbindet den Austausch von Wasser zwischen Fluss und Grundwasser. Bei Hochwasser kann die Kolmationsschicht wieder aufgerissen werden. Falls der Grundwasserspiegel über der Flusssohle oder, je nach Grad ihrer Kolmatierung, höchstens wenige Dezimeter darunter liegt, infiltriert der Fluss direkt ins wassergesättigte Sediment (z. B. Bouwer 1978, Hoehn et al. 1983). Der hydraulische Anschluss des Flusses an das Grundwasser unterbindet die Zufuhr von Bodenluft. Bei tiefer liegendem Grundwasserspiegel geht der hydraulische Anschluss verloren und es entsteht eine ungesättigte Zone zwischen dem durchlässigen Flussbett und dem Grundwasserspiegel. Bei einer solchermaßen indirekten Infiltration versickert Flusswasser durch eine ungesättigte und belüftete Zone hindurch ins Grundwasser. Die ungesättigte Zone weist häufig eine wesentlich geringere Durchlässigkeit auf als die gesättigte. Bei Infiltrationsbedingungen besteht das Grundwasser aus einer Mischung von verschiedenen Typen: i) flussnahem Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 Infiltrat (hyporheischem Grundwasser), ii) älterem Infiltrat von weiter flussaufwärts, sowie iii) Grundwasser aus der Versickerung von Niederschlagswasser. Unter flussnahem Infiltrat verstehen wir Wasser, welches eine Aufenthaltszeit im Untergrund von < 10 d bzw. < 50 d aufweist, was dem Wert für die äußere Begrenzung der Zone 2/II in der Schweiz bzw. in Deutschland entspricht. Die drei Grundwassertypen treten im Grundwasserleiter oft geschichtet auf und sind chemisch unterschiedlich zusammengesetzt (Abb. 1; z. B. Hoehn et al. 2007). Zwischen dem Fluss und dem Auftreten an einer Messstelle weisen die verschiedenen Grundwassertypen unterschiedlich lange Verweilzeiten im Untergrund auf. Maßgeblich für die Trinkwasserqualität in einer Fassung ist die Verweildauer des Anteils an flussnahem Infiltrat und nicht jene des geförderten Mischgrundwassers. In alpenrandnahen Schotterebenen betragen die Verweilzeiten des flussnahen Infiltrats bis zur Trinkwasserfassung oft nur wenige Tage. Derart kurze Verweilzeiten können unter gewissen Bedingungen bestimmt werden, z. B. mittels • ereignisgesteuerten Zeitreihenanalysen (bis max. 30 d; Cirpka et al. 2007, Vogt et al. 2009), • Markierversuchen, 257 • Bestimmung des Radon-Wasseralters (bis max. 15 d; Hoehn & von Gunten 1989) und • Bestimmung des 3H-3He-Wasseralters (ab 60 d; z. B. Beyerle et al. 1999). Erstere führen zu kontinuierlichen Verteilungen der Aufenthaltszeiten, letztere ergeben ein Mischalter der Wasserprobe. Tabelle 1 zeigt Ereignisse, die zu einer Verminderung der Verweildauer bzw. zu einer Erhöhung des Anteils an flussnahem Infiltrat führen können. Die Anteile der einzelnen Grundwassertypen im geförderten Mischgrundwasser können quantifiziert werden. Grundwasser aus versickerten Niederschlägen ist meist stärker mineralisiert und härter als flussnahes Infiltrat (z. B. Hoehn et al. 1983). Im voralpinen Raum ist die Isotopensignatur (z. B. δ18O; Stichler et al. 1986) ein gutes Maß für die Abschätzung des Anteils an flussnahem Infiltrat. Viele Flüsse enthalten Anteile an (geklärten und ungeklärten) Abwässern. Dies verschlechtert die Qualität des Flusswassers und wirkt sich entsprechend auf die Qualität des Infiltrats aus (z. B. Bourg & Bertin 1994). In der dicht besiedelten Schweiz weist flussnahes Infiltrat Anzeichen einer Belastung mit persistenten organischen Verbindungen aus Abwässern auf (z. B. Hoehn et al. 1983). Älteres Infil Abb. 1 Schematische Darstellung des Infiltrationsvorgangs, mit Grundwasserströmung zu einem flussnahen Brunnen: 1: Flusswasser; 2: flussnahes Infiltrat; 3: älteres, weiter stromaufwärts infiltriertes Grundwasser; 4: Grundwasser aus versickerten Niederschlägen, Hangwasser; 5: Wasser aus Anlage für künstliche Grundwasseranreicherung; A: stark gefährdeter unvollkommener Brunnen, mit flussnahem Infiltrat; B: gefährdeter vollkommener Brunnen, mit Mischgrundwasser; C: gelegen zwischen Fluss und Brunnen B (siehe auch Tab. 2) (verändert, aus Hoehn et al. 2007) 123 258 Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 Tabelle 1 Beispiele von Ereignissen in und an Fließgewässern und ihre Auswirkungen auf das in nahe gelegenen Fassungen mit Infiltratanteil geförderte Grundwasser Ereignis Auswirkung 1 Hochwasserwellen • Dekolmation des Fließgewässerbetts des Flusses • Erhöhung der spezifischen Infiltrations rate (Schluckvermögen) • Erhöhung des Anteils an flussnahem Grundwasser im geförderten Grundwasser • Reduktion der Verweildauer des geförderten Mischgrundwassers 2 Lange Trocken • Kolmation des Fließgewässerbetts perioden • Erniedrigung der spezifischen Infiltrationsrate (Schluckvermögen) • Erniedrigung des Anteils an flussnahem Grundwasser im geförderten Grundwasser • Erhöhung der Verweildauer des geförderten Mischgrundwassers 3 Große Schwankun- • Bei indirekter Infiltration: Verkleinegen des Grundwasrung der Distanz zwischen Flusssohle serspiegels und Grundwasserspiegel, bis hin zur Möglichkeit des Auftretens von direkter Infiltration • Wechsel zwischen Exfiltration und direkter Infiltration, was im unteren Teil von Schotterebenen auftritt trat hingegen ist in seiner Zusammensetzung oft nur noch schwach von Grundwasser aus versickerten Niederschlägen zu unterscheiden. Falls das Infiltrat einen Gehalt an organischem Kohlenstoff von deutlich über 1 µg/g aufweist, kann die mikrobiell mediierte Sauerstoffzehrung, v. a. bei sommerlichen Wassertemperaturen, bedeutend werden (von Gunten et al. 1991). Dies kann bei direkter Infiltration belasteter Flüsse in gewissen Fällen zur Eisenlösung führen. Bei Flussraum-Aufweitungen im Anströmbereich von Fassungen mit flussnahem Infiltratanteil nimmt die Gefährdung noch zu. In den meisten Ländern Europas soll Grundwasser möglichst ohne Aufbereitung als Trinkwasser genutzt werden können. Das geförderte Rohwasser soll chemisch und mikrobiologisch so beschaffen sein, dass die gesetzlich festgelegten Trinkwasserwerte eingehalten werden. Zudem sollen Anlagen und Tätigkeiten das Grundwasser nicht beeinträchtigen. In vielen voralpinen und alpinen Schotterebenen befinden sich bedeutende Trinkwasserfassungen in der Nähe von Flussläufen, weil dort das Dargebot an Grundwasser in der Regel größer und beständiger ist als in größerer Distanz zum Fluss. Diese Fassungen fördern zu einem nennenswerten Anteil Flussinfiltrat. Für den Infiltratanteil solcher Fassungen reicht der Entnahmebereich bis ans Flussufer. Das geförderte Wasser weist wegen der Verdünnung durch das Infiltrat oft eine geringere Belastung durch Nitrat und Pflanzenschutzmittel auf als Grundwasser aus Niederschlägen. 123 Zudem ist es im Normalfall frei von pathogenen Bakterien, denn diese werden in sauberem Kies-Sand relativ rasch eliminiert (z. B. Kustermann 1962; coliforme Keime im Laborversuch: > 90 % innerhalb von 10 Tagen, siehe Merkli 1974; neuere Literatur zusammengefasst in Regli & Huggenberger 2007). Inhaber von Trinkwasserfassungen sind verpflichtet, Wasserschutzgebiete einzurichten (Deutschland: Wasserhaushaltsgesetz, § 19; siehe DVGW 2006) bzw. Schutzzonen auszuweisen (Schweiz: Gewässerschutzgesetz, Art. 20; BUWAL 2004). Kritische Parameter für die Wasserqualität flussnaher Fassungen sind i) der Infiltratanteil, ii) die Verweilzeiten des Infiltrats und iii) die Qualität des Flusswassers. In Deutschland gilt die Uferfiltration vielerorts als erste von mehreren Aufbereitungsstufen für Trinkwasser (z. B. Sontheimer & Nissing 1977, Tufenkji et al. 2002). Idealerweise liegen Fassungen in Distanzen zum Fluss, die es erlauben, das Grundwasser ohne Aufbereitung zu nutzen. Innerhalb der Engeren Schutzzone (Zone II bzw. Zone S2) um eine Trinkwasserfassung herum muss das geförderte Grundwasser eine Verweildauer von mindestens 50 Tagen (DVGW 2006) bzw. 10 Tagen in der Schweiz (Gewässerschutzverordnung, siehe auch BUWAL 2004) aufweisen. Fassungen mit Infiltratanteil sind generell schwieriger zu schützen als Fassungen mit Grundwasser aus Niederschlägen (Regli et al. 2003). Der Infiltratanteil muss auf der Strecke zur Fassung genügend gut gereinigt werden. Wenn dieser weitgehend Salze sowie gut abbaubare und sorbierende Stoffe aus geklärten Abwässern enthält, bestehen keine nennenswerten Probleme mit der Grundwassernutzung zu Trinkwasserzwecken. In industrialisierten Gebieten enthalten geklärte Abwässer jedoch zusätzlich Spuren von nicht abbaubaren, mobilen und, falls in höheren Konzentrationen vorliegend, teilweise toxischen Schadstoffen. Diese können mit dem Infiltrat bis zu den flussnahen Trinkwasserfassungen gelangen. Deshalb stellen wir im Folgenden ein Beurteilungssystem bezüglich des Risikos durch FlussraumAufweitungen zur Diskussion und schlagen Maßnahmen zur Minderung der Gefährdung vor. Methodischer Ansatz für die Beurteilung flussnaher Grundwasserfassungen In den letzten Jahren wurden Fälle bekannt, bei denen Trinkwasserfassungen, welche Wasser mit nennenswertem Infiltrat-Anteil fördern, nach Flussraum-Aufweitungen Qualitätseinbußen erlitten. Die Verweildauer des geförderten Mischgrundwassers nahm ab und fiel unter die geforderten Werte. Zudem nahm der Anteil an flussnahem Infiltrat zu (SVGW 2007). Als methodischen Ansatz für die Beurteilung der Gefährdung flussnaher Grundwasserfassungen Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 durch Flussraum-Aufweitungen schlagen wir vor, folgende hydraulische Kriterien zu verwenden: i) Aufenthaltszeiten des flussnahen Infiltrats, ii) dessen Mischungsanteil im geförderten Wasser und iii) das Vorhandensein oder Fehlen einer ungesättigten Zone zwischen Flusssohle und Grundwasserspiegel. Gegenüber jenen von Regli & Huggenberger (2007) und des SVGW (2007) ist der vorliegende Ansatz insofern verschieden, als er weder von der Reinigungsleistung der Flusssohle noch von der stofflichen Belastung des Fließgewässers abhängt. Die Anwendbarkeit des hier zur Diskussion gestellten Ansatzes wird im Folgenden mit Fallbeispielen aus voralpinen Schotterebenen der Schweiz unterlegt. Bei der Gefährdung der Trinkwasserversorgung als Folge von Flussraum-Aufweitungen unterscheiden wir drei Fälle: i) Generell geringes Risiko, ii) Risiko während oder nach durchgeführten FlussraumAufweitungen, iii) hohes Risiko bei problembehafteten Fassungen, unabhängig von geplanten Flussraum-Aufweitungen (Problemfälle). Wenn die vorgängig beschriebenen Grundlagen quantifiziert vorliegen, kann das Risiko einer Beeinträchtigung der Fassung mit diesem methodischen Ansatz abgeschätzt werden. Fälle mit geringem Risiko Das Risiko einer Verunreinigung des Grundwassers in einer Fassung infolge Flussraum-Aufweitungen ist nicht vorhanden oder gering, falls im aufzuwertenden Flussabschnitt dauernd und ausnahmslos • Beziehungen zwischen Fließgewässer und Grundwasser fehlen, d. h. das Flussbett verläuft natürlicherweise in gering wasserdurchlässigen Deckschichten, oder • das Grundwasser ins Fließgewässer exfiltriert, d. h. die Fließrichtung des Grundwassers sowohl bei niedrigen Grundwasserspiegeln als auch bei Dauerförderung mit der konzessionierten Pumpleistung entweder in Richtung Fließgewässer oder zumindest talparallel verläuft (z. B. im Thurtal, siehe Vogt et al. 2009), oder • der Entnahmebereich des Fließgewässers entweder unterhalb der geplanten Flussraum-Aufweitungen liegt oder stromaufwärts in einer Distanz, welche direkte Fließwege zur Fassung ausschließt. Unter diesen Bedingungen ist die Sicherheit der Trinkwasserversorgung auch dann gewährleistet, wenn das Ufer des Fließgewässers nach einer Aufweitung näher bei der Fassung liegt. Eine Kolmation der Flusssohle über durchlässigem Gestein kann die Beziehungen zwischen Fluss und Grundwasser vermindern und im Extremfall ganz unterbinden. In diesem Fall strömt das Grundwasser unter dem Fluss hindurch, ohne mit flussnahem Infiltrat angereichert zu werden. 259 Während Flussraum-Aufweitungen in der Nähe von Trinkwasserfassungen muss sichergestellt werden, dass solche dichtende Schichten nicht durchbrochen werden, weil sonst zwischen Fließgewässer und Grundwasser ein unerwünschter Austausch erzeugt werden kann. Wenn der Fluss auf künstlich abgedichtetem Bett verläuft, ist nach einer Uferaufweitung damit zu rechnen, dass er ins Grundwasser infiltriert. Aufweitungen in der Nähe von Fassungen dürfen das Fließgewässer nicht aufstauen, damit der Infiltratanteil in der Fassung nicht zunimmt. Auch darf das Grundwasser nicht abgesenkt werden, z. B. mittels Entwässerungskanälen. Als Beweismittel und Wirksamkeitskontrolle sind während einer Zeit von minimal einem Jahr vor und während der Flussraum-Aufweitung folgende Parameter in angepasster Frequenz zu überwachen und ihre Werte mittels Analysen von Zeitreihen auszuwerten (BUWAL 2004): • Abstand zwischen Flusssohle und Grundwasserspiegel, • Verweildauer im Untergrund bzw. Alter des geförderten Grundwassers, • Mischanteile des geförderten Grundwassers, • Chemischer und mikrobiologischer Zustand des Fließgewässers und des geförderten Grundwassers. Zudem sollen Extremsituationen (z. B. Hochwasserspitzen) ereignisorientiert überwacht werden. Der Aufstau von Flüssen bei Laufkraftwerken hat gezeigt, dass es bis zum Aufbau einer stabilen Kolmationsschicht an der Flusssohle Jahre dauern kann (z. B. Limmat, Wettingen: Harder 1935, 1938). Mangels genügend langer Erfahrungen über Veränderungen in der Grundwasserqualität als Folge von Flussraum-Aufweitungen sollte deshalb die Überwachung nach dem Eingriff unseres Erachtens minimal drei Jahre weitergeführt werden. Fälle mit Risiko während oder nach durchgeführten Flussraum-Aufweitungen Insbesondere bei Fließgewässern, welche ins Grundwasser infiltrieren, ist bei einer Flussraum-Aufweitung mit erheblichen Auswirkungen auf das Grundwasser und daher mit dem Risiko einer Verunreinigung von Trinkwasserfassungen zu rechnen. Die WRRL interpretiert ihre Ziele vorab in ökotoxikologischer und strukturmorphologischer Hinsicht. Bei der Umsetzung der im Entwurf vorliegenden Tochterrichtlinie über Grundwasser wurden die Anliegen der Fachleute für eine einwandfreie Versorgung mit Trinkwasser nur ungenügend berücksichtigt (EU 2006). Dies kann sich insbesondere für flussnahe Fassungen nachteilig auswirken. Die derzeitig geforderten Werte berücksichtigen nicht, dass trinkwasserrelevante Stoffe, die sich in der Umwelt als schlecht abbaubar erweisen, den natürlichen Wasserkreislauf nicht belasten sollten (Wirtz 2006). In Deutschland regelt das Arbeitsblatt W 101 (DVGW 2006) den Fall von 123 260 Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 Uferfiltrat und Seihwasser in Kapitel 4.5.4, in dem Sinn, dass geprüft werden muss, ob zumindest Teile des infiltrierenden oberirdischen Gewässers und seines Einzugsgebiets ins Trinkwasserschutzgebiet einbezogen werden sollen. Andernfalls wäre die Reinhaltung dieses Gebiets durch andere Maßnahmen zu gewährleisten. Für flussnahe Fassungen, welche einen erheblichen Anteil Infiltrat fördern, sind oberirdische Gewässer, die mit Abwasser belastet sind, problematisch. In der Schweiz sind wasserbauliche Maßnahmen für die Fließgewässer-Revitalisierung nur außerhalb der Engeren Schutzzone erlaubt und in der Weiteren Schutzzone S3 bewilligungspflichtig (BUWAL 2004). Bei Fassungen mit einem geringen Anteil an flussnahem Infiltrat dürften kurzfristige Überschreitungen von gesetzlich geforderten Werten von Schadstoffen im Fluss durch Misch-, Verzögerungs- und evtl. Abbauvorgänge in den meisten Fällen genügend stark gedämpft werden, um die Anforderungen an Trinkwasser zu erfüllen. Für die Beurteilung des Risikos von Flussraum-Aufweitungen im Einflussbereich flussnaher Trinkwasserfassungen ist zu unterscheiden zwischen Flüssen, die indirekt über eine ungesättigte Zone ins Grundwasser infiltrieren, und solchen mit direktem Anschluss ans Grundwasser (Regli & Huggenberger 2007). Bei einer dauernd indirekten Infiltration ist das Risiko einer Verunreinigung der Trinkwasserfassung eher gering. Ebenso bleibt bei einer geringen Durchlässigkeit der ungesättigten Zone der Anteil an flussnahem Infiltrat eher tief und das Risiko der Beeinträchtigung einer flussnahen Fassung eher klein. Im Folgenden stellen wir für den Fall von indirekter Flussinfiltration in alpinen und voralpinen Schottergebieten Bedingungen zur Diskussion, die aus Sicht der Wasserversorgung idealerweise erfüllt sein sollten, und zwar sowohl bei der Planung von Flussraum-Aufweitungen als auch nach den ausgeführten Arbeiten. Unser Vorschlag knüpft an die Erfahrungen bei der Modellierung der Auswirkungen geplanter Flussraum-Aufweitungen im oberen Bereich des Thurtals an (Bürglen-Weinfelden: Baumann 2009; Schachen-Weinfelden: Vogt et al. 2009). Dort werden zwar deutliche Erhöhungen des Grundwasserspiegels erwartet, die aber nicht zu wesentlich kürzeren Aufenthaltszeiten zu Fassungen führen dürften. i) Die Fassung muss sich in einer Distanz vom Ufer weg befinden, die derart groß ist, dass das flussnahe Infiltrat im gesättigten Bereich des Grundwasserleiters die geforderte Verweildauer von mindestens 10 bzw. 50 Tagen aufweist (Engere Schutzzone). Neben der Selbstreinigung gegenüber pathogenen Mikroorganismen, gewährleistet die geforderte minimale Verweildauer auch eine ausreichende Vorwarnzeit bei unfallbedingten Verunreinigungen des Flusswassers. ii) Im ungesättigten Bereich des Grundwasserleiters mit kiessandigem Material zwischen Flusssohle und Grundwasserspiegel sollte sich eine Zone mit einer Mäch- 123 tigkeit von mindestens 10 m über dem natürlichen, zehnjährigen Grundwasserhöchstspiegel befinden. In vorsichtiger Interpretation des Gesetzes von Darcy wird angenommen, dass eine spezifische Infiltrationsrate, q, von 10 m3 m–2 d–1, eine nutzbare Porosität für Wasser im ungesättigten Bereich (volumetrischer Wassergehalt), p, von 10 %, und eine Potenzialdifferenz, x, von 10 m zwischen Flusssohle und Grundwasserspiegel zu einer Verweildauer, t, von t = x · p/q = 10 d führt. iii) Der Anteil an flussnahem Infiltrat sollte 10 % nicht übersteigen. Damit werden kurzfristige hohe Belastungen von Schad- oder Fremdstoffen im Flusswasser durch Mischungs-, Verzögerungs- und evtl. Abbauvorgänge in der Regel genügend stark gedämpft, sodass die Trinkwassergewinnung nicht gefährdet wird. Unser Vorschlag befindet sich bewusst auf der vorsichtigen Seite: Die Erfahrungen im Thurtal zeigten, dass Mischungsrechnungen über die Endglieder Flusswasser und Grundwasser aus Niederschlägen, z. B. mittels Alkalinität oder Chlorid, bei geringen Konzentrationsunterschieden nur wenig genau sind (Hoehn et al. 2007). Falls alle drei Bedingungen (> 10 m mächtige ungesättigte Zone bzw. 10 d Verweilzeit des Sickerwassers in der ungesättigten Zone, minimal für die Engere Schutzzone geforderte Verweildauer des Grundwassers im gesättigten Grundwasserleiter und < 10 % Anteil an flussnahem Infiltrat) erfüllt sind, besteht unseres Erachtens aus Sicht der Wasserversorgung ein gewisser Spielraum für Flussraum-Aufweitungen. Im kritischeren Fall einer zumindest zeitweiligen direkten Infiltration in den gesättigten Grundwasserleiter besteht höchstens dann ein gewisser Spielraum für Flussraum-Aufweitungen, wenn gezeigt werden kann, dass die Verweilzeiten im heutigen Zustand deutlich über den geforderten minimalen Werten liegen und der Anteil an flussnahem Infiltrat unter dem vorgeschlagenen Wert von 10 % liegt. Das dürfte vor allem in alpinen und voralpinen Schotterebenen nur selten der Fall sein. Bei solchermaßen gefährdeten Trinkwasserfassungen müssten unseres Erachtens die weiter oben erwähnten Überwachungsmaßnahmen auf eine Zeit von minimal 2 Jahren vor, während, und minimal sechs Jahren nach den Aufweitungen – bzw. bis sich die Flusssohle stabilisiert hat – ausgedehnt werden. Gegenüber den Fällen mit geringem Risiko werden hier die Überwachungsmaßnahmen verschärft. Mit dieser Verschärfung beziehen wir uns auf Fälle von Verschlechterungen der Grundwasserqualität als Folge von Flussraum-Aufweitungen. Das Aufbrechen der bestehenden Uferverbauungen im Fluss Wiese bei Basel („Lange Erlen“) führte in einer Fassung der Industriellen Werke Basel in einer Distanz vom revitalisierten Ufer von ca. 140 m zu einer ursächlichen Verringerung der Aufenthaltszeit um etwa einen Faktor von 2 und einer regelmäßigen Verkeimung. Diese Fassung wurde sicherheitshalber vom Netz entfernt (Hug- Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 genberger 2001, SVGW 2007). In einer Auenlandschaft der Aare zwischen Bern und Thun wurde ein Grundwasserwerk und – während des Baus der Fassung – als Revitalisierung ein Infiltrationsgraben sowie Teiche und Feuchtgebiete als „Seitenarm“ der Aare erstellt. Dies führte zu einem Durchbruch von Kleinlebewesen und sporadisch auftretenden bakteriologischen Verunreinigungen. Erst als sowohl der Graben als auch der Seitenarm wieder geschlossen wurden, entsprach die Qualität des geförderten Wassers den gesetzlichen hygienischen Bestimmungen (SVGW 2007). Bereits bei der Planung von Flussraum-Aufweitungen sind Maßnahmen zum Schutz flussnaher Grundwasserfassungen vor Verunreinigung vorzusehen, z. B.: • Bezeichnung einer Distanz vom heutigen Ufer aus für die größte zugelassene Aufweitung des Flussraums. Dies ergibt Zeit für Planung und Durchführung von Gegenmaßnahmen, für den Fall, dass der Fluss sich nach einer Aufweitung weiter in seitlicher Richtung ausdehnen sollte (Baumann 2003). • Verstärktes Aufteilen der vorgesehenen Eingriffe in Etappen, z. B. auf einzelne kleinere Flussabschnitte (Oplatka 1999). Dies erlaubt, Folgen abzuschätzen und allenfalls Gegenmaßnahmen einzuleiten. 261 • Erarbeitung von Alternativlösungen mit weniger starken Auswirkungen auf das Grundwasser in der Nähe von Trinkwasserfassungen (Baumann et al. 2009, Baumann 2003). Dies wird nötig, falls eine Modellierung der Infiltrationsverhältnisse unerwünscht starke Auswirkungen vorhersagt (z. B. Anstieg der Grundwasserspiegel). Mögliche Hochwasserwellen des Flusses und große Schwankungen des Grundwasserspiegels können bewirken, dass eine oder mehrere der oben beschriebenen Bedingungen nicht erfüllt sind, und daher der notwendige Schutz der Wasserversorgung nicht gewährleistet werden kann (Tab. 1). In solchen Fällen ist grundsätzlich auf wasserbauliche Eingriffe zu verzichten, die über die unbedingt erforderlichen Hochwasserschutz-Maßnahmen hinausgehen. Die zuständigen Behörden sollten Bewilligungen für solche Maßnahmen nur nach besonders sorgfältiger Prüfung und mit vorsorglichen Schutzmaßnahmen für die Trinkwasserfassungen erteilen. Innerhalb der Engeren Schutzzone sind Flussraum-Aufweitungen nur dann zulässig, wenn eine Gefahr für die Trinkwassernutzung ausgeschlossen werden kann und sie bei Hochwassergefahr zur Rettung von Menschenleben oder erheblichen Sachwerten unerlässlich sowie standortgebunden sind. Wo auf Aufweitungen innerhalb Tabelle 2 Beispiele technischer Maßnahmen an problembehafteten Grundwasserfassungen mit Infiltratanteil und ihrer Umgebung zur Minderung des Risikos einer Grundwasser-Verunreinigung Maßnahme Auswirkung 1 Generelle Drosselung der Grundwasserförderung 2 3 4 5 6 Bei einer Drosselung der Grundwasserförderung wird der Absenktrichter rund um die Fassung herum kleiner. Damit erhöht sich die Verweildauer, und der relative Anteil an flussnahem Infiltrat verringert sich. Eine Drosselung kann z. B. über eine Verringerung der konzessionierten Wassermenge erreicht werden. Abhängigkeit der Grundwas- Mit dem zeitweiligen Ausschalten der Fassung oder zumindest starken Drosselung der Förderleistung bei serförderung vom Abfluss des und unmittelbar nach Hochwasserwellen in Fließgewässern kann der hochwasserbedingten Verringerung der Verweilzeiten und Erhöhung des Infiltratanteils (Gefälle, Schluckvermögen) entgegengewirkt werden. Fließgewässers Das Pumpregime kann in der Grundwasserfassung wasserstands- oder abflussabhängig gesteuert werden. Einzug von Vollrohren im obe- Falls sich das flussnahe Infiltrat oben im Grundwasserleiter einschichtet, könnte der Anteil an flussnahem ren Bereich des Filterbrunnens Infiltrat im geförderten Grundwasser vermindert werden, wenn nur noch aus größeren Tiefen gepumpt wird. Künstliche Abdichtung von Durch künstliche Abdichtung der Uferzonen kann die Infiltration von Oberflächenwasser in das GrundwasUferzonen des Fließgewässers ser unterbunden werden. Dazu müssen Abdichtungen bis an die Basis des wasserdurchlässigen Kiessands abgeteuft werden, weil sonst das flussnahe Infiltrat den Abdichtungskörper unter- bzw. umströmt. Bei kleinen Fließgewässern mit ungenügender Wasserqualität können die Sohlen abgedichtet werden. Spundwände sind im Kiessand richtungsstabil und genügend gut abdichtend; Schlitzwände sollten nur mit Bentonit als Dichtungsmasse ausgeführt werden, d. h. ohne organische Zusatzstoffe. Beispiele von Abdichtungen beim Aufstau von Flüssen zeigen, dass diese Maßnahme über viele Jahre hindurch wirksam ist (z. B. Rheinaufstau Kraftwerk Säckingen, Aareaufstau Kraftwerk Wildegg-Brugg; Jäckli & Kempf 1972). Nachteil vor allem von Dichtwänden ist, dass es sich um irreversible Eingriffe in den Grundwasserleiter handelt. Abdichtungen sind massive bauliche Eingriffe in den Untergrund, deren Durchführung spezieller Rechtfertigung bedarf. Vorsorgliche Wasseraufberei- Der vorsorgliche Einbau einer Wasseraufbereitung ist eine anerkannte Maßnahme. Das Signal für das tung Zuschalten der Aufbereitung kann z. B. über die Abflussmenge des infiltrierenden Fließgewässers, der Lage des Grundwasserspiegels oder über die Trübung gesteuert werden. Künstliche Anreicherung Wenn das in der Fassung geförderte Mischwasser infolge zu hohen Anteils an flussnahem Infiltrat eine zu kurdes Grundwassers ze Verweildauer aufweist, kann die Möglichkeit erwogen werden, eine Anlage zur künstlichen Anreicherung des Grundwassers zu erstellen. Diese darf (anders als im Vorschlag von Regli et al. 2003) aber nicht zwischen Fließgewässer und Pumpwerk gebaut werden, weil so das Mischalter infolge der sehr geringen Verweildauer des Anreicherungswassers noch kleiner wird (Abb. 1). Wasser müsste landseitig in größerer Distanz von der Fassung weg zur Versickerung gebracht werden. Im Extremfall kann damit die Fließrichtung des Grundwassers so verändert werden, dass kein flussnahes Infiltrat mehr aus dem Fließgewässer in die Fassung gelangt. 123 262 Grundwasser – Zeitschrift der Fachsektion Hydrogeologie (2009) 14:255–263 der Engeren Schutzzone nicht verzichtet werden kann, sind Schutzmaßnahmen für das Grundwasser vorzusehen (Regli et al. 2003, Regli & Huggenberger 2007). Erst wenn diese Möglichkeiten nicht realisiert werden können, wäre eine Verlegung der Fassung bzw. die Beschaffung von Ersatzwasser zu prüfen. Im Fall der Vorbereitungen für die geplanten FlussraumAufweitungen im Rahmen der 3. Thurkorrektion wurden gefährdete Trinkwasserfassungen untersucht. Der in Cirpka et al. (2007) und Hoehn et al. (2007) beschriebene Fall der Fassung Widen, Frauenfeld, ist beispielhaft: Minimale Aufenthaltszeiten bei und nach Hochwasserwellen von knapp 10 Tagen ließen es angebracht erscheinen, in diesem Flussabschnitt auf Aufweitungen zu verzichten. Als Folge der erwähnten Erfahrungen müssen den Wasserversorgern technische Maßnahmen zur Minderung des Risikos einer Beeinträchtigung von Fassungen mit hohem Anteil an flussnahem Infiltrat aufgezeigt werden können. Solche Maßnahmen schlagen wir in Tabelle 2 vor. Deren Auswirkungen auf die Verweildauer des Grundwassers und den Infiltratanteil müssen vorgängig mit einem mit Felddaten gespeisten instationären Strömungsmodell simuliert und vorhergesagt werden. Mit den Maßnahmen aus Tabelle 2 können unter gewissen Umständen unerwünschte Strömungssituationen verbessert werden. Die Tabelle soll aber nicht den Eindruck einer übermäßigen Machbarkeit erwecken. Jede Fassung ist ein Einzelfall, der mit den geometrischen, geologischen und hydrologischen Parameterwerten beurteilt werden muss. Problembehaftete Fassungen Problembehaftete Fassungen weisen auch ohne FlussraumAufweitungen zumindest zeitweise zu geringe Verweilzeiten bis zur Entnahme im Pumpwerk (< 10 bzw. < 50 d; Engere Schutzzone reicht bis ans Ufer) und/oder zu hohe Infiltratanteile verunreinigter Flüsse (> 10 %) auf. Dieser Fall ist vielerorts mit einem direkten hydraulischen Anschluss des Grundwassers ans Fließgewässer verbunden. Bei schlechter Fließgewässerqualität, z. B. bei Stoßbelastungen durch ungenügend geklärte Abwässer oder Regenentlastungen, erhöht sich das Risiko einer Grundwasserverunreinigung deutlich. In diesen Fällen sollte auf den Zeitpunkt des Ablaufs der Konzession zur Entnahme von Grundwasser zu Trinkwasserzwecken ein Fassungsstandort an weniger exponierter Lage gefunden oder Ersatzwasser beschafft werden. Ersteres ist in dicht besiedelten Gebieten schwierig zu realisieren, wo neben den Fließgewässern viele andere raumwirksame Nutzungen mit dem Grundwasserschutz kollidieren. Das Risiko der Verunreinigung einer Grundwasserfassung mit (zu) hohem Infiltratanteil sollte bis zum Ablauf der Konzession mit planerischen und technischen Maßnahmen reduziert werden können. 123 Ausblick Zusammenfassend beurteilen wir Trinkwasserfassungen mit flussnahem Infiltratanteil nach ihrem Verunreinigungsrisiko bei Flussraum-Aufweitungen. Wir beschränken uns dabei auf hydraulische Beurteilungsgrößen und vermeiden einen Ansatz mit Berücksichtigung der stoffspezifischen Gefährdung. Damit glauben wir, einen Beitrag zur besseren Identifizierung solchermaßen gefährdeter Fassungen zu leisten. Aufweitungen des Fließgewässerraums für den Schutz gegen Hochwasser bzw. zur Aufwertung des Gewässers geraten unter Umständen in einen Konflikt mit dem Grundwasserschutz, was Planer und Vollzugsbehörden vor große Herausforderungen stellt. Um eine Grundwasserfassung vor zu hohen Anteilen an flussnahem Infiltrat und zu geringer Verweildauer zu schützen, müssen die Fließwege und die Fließdynamik im Entnahmebereich einer Fassung vor Beginn der Arbeiten gut bekannt sein. Die zahlenmäßige Bestimmung von Verweilzeiten und Infiltratanteil setzt gute Kenntnisse der Durchlässigkeit und der Grundwasserströmung (Fließrichtung und -geschwindigkeit) bei Niedrig-, Mittel- und Hochwasser voraus. Die hydrogeologische Erkundung einer Fassung mit einem nennenswerten Anteil an flussnahem Infiltrat umfasst gegenüber einer solchen für Grundwasserfassungen in einem rundum ausgedehnten Grundwasserleiter ein Mehrfaches an Aufwand. Erst bei genauer Kenntnis der Mischungs- und Strömungssituation kann abgeschätzt werden, ob eine schutzbedürftige Grundwasserfassung auch nach Flussraum-Aufweitungen noch ordnungsgemäß geschützt werden kann. Verweilzeiten und Mischanteile von Grundwasser als Größen für eine Beurteilung von Flussinfiltration dürften sich sinngemäß auch bei der künstlichen Anreicherung von Grundwasser verwenden lassen. Erwünscht sind auch dort belastbare Zahlenwerte, und diese sind mit der Qualität des eingespeisten Rohwassers und dem vorgesehenen Aufwand für die Aufbereitung des geförderten Wassers zu verknüpfen. Danksagung Wir profitierten von Ergebnissen der Arbeitsgruppe „Fluss-Grundwasser-Interaktion“ der Schweizerischen Gesellschaft für Hydrogeologie (Leitung: P. Huggenberger). Wir danken den Herren Daniel Hartmann und Reto Muralt, beide Sektion Grundwasserschutz, Bundesamt für Umwelt (BAFU), für die Durchsicht des Manuskripts. Literatur Baumann, M., Hoehn, E., Jordan, P.: Die 2. Thurgauer Thurkorrektion erfordert einen neuen Blick auf das Thurtaler Grundwasser.- Mitt. Thurgauer natf. 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https://openalex.org/W3165668011	OpenAlex	Open Science	CC-By	2,021	Research models and mesenchymal/epithelial plasticity of osteosarcoma	Xiaofeng Yu	English	Spoken	10,799	21,214	© The Author(s) 2021. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativeco mmons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/ zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Research models and mesenchymal/ epithelial plasticity of osteosarcoma Xiaobin Yu1, Jason T. Yustein2 and Jianming Xu1* Abstract Most osteosarcomas (OSs) develop from mesenchymal cells at the bone with abnormal growth in young patients. OS has an annual incidence of 3.4 per million people and a 60–70% 5-year surviving rate. About 20% of OS patients have metastasis at diagnosis, and only 27% of patients with metastatic OS survive longer than 5 years. Mutation of tumor suppressors RB1, TP53, REQL4 and INK4a and/or deregulation of PI3K/mTOR, TGFβ, RANKL/NF-κB and IGF pathways have been linked to OS development. However, the agents targeting these pathways have yielded disappointing clinical outcomes. Surgery and chemotherapy remain the main treatments of OS. Recurrent and metastatic OSs are commonly resistant to these therapies. Spontaneous canine models, carcinogen-induced rodent models, transgenic mouse models, human patient-derived xenograft models, and cell lines from animal and human OSs have been developed for studying the initiation, growth and progression of OS and testing candidate drugs of OS. The cell plas- ticity regulated by epithelial-to-mesenchymal transition transcription factors (EMT-TFs) such as TWIST1, SNAIL, SLUG, ZEB1 and ZEB2 plays an important role in maintenance of the mesenchymal status and promotion of cell invasion and metastasis of OS cells. Multiple microRNAs including miR-30/9/23b/29c/194/200, proteins including SYT-SSX1/2 fusion proteins and OVOL2, and other factors that inhibit AMF/PGI and LRP5 can suppress either the expression or activity of EMT-TFs to increase epithelial features and inhibit OS metastasis. Further understanding of the molecular mechanisms that regulate OS cell plasticity should provide potential targets and therapeutic strategies for improving OS treatment. Keywords: Osteosarcoma, Metastasis, Experimental model, EMT/MET-related process, EMT-TFs OS arises from primitively transformed cells with a mesenchymal origin [4]. The cancer cells in OS look like early forms of bone cells that normally help gener- ate new bone tissues, but the bone tissues in OS are not as strong as that of normal bones [5]. Each year, about 800 to 900 new cases of OS are diagnosed in the United States [5]. Based on patient ages and causes of OS devel- opment, they can be classified into primary and second- ary OSs. The primary OS typically develops in young patients as a result of abnormal bone development. The secondary OS occurs in patients over 65 years old, and is usually secondary to malignant conditions of Paget’s disease, post-irradiation exposure, severe bone infarct, osteochondroma and osteoblastoma [6]. Based on the location and appearance, OSs can be classified into Yu et al. Cell Biosci (2021) 11:94 https://doi.org/10.1186/s13578-021-00600-w Yu et al. Cell Biosci (2021) 11:94 https://doi.org/10.1186/s13578-021-00600-w Cell & Bioscience Cell & Bioscience Background Osteosarcoma (OS) is the most common type of cancer that initiates at the bone, with a worldwide incidence of 3.4 per million people each year [1]. The 5-year sur- vival rate for classic OS was only 20% during most of the twentieth century until the introduction of adjuvant chemotherapy in the 1970s [2]. The routine treatment of high-grade OS also shifted from amputation to chemo- therapy and limb salvage by 1990, with a subsequent increase in overall survival rate to more than 65% [3, 4]. *Correspondence: jxu@bcm.edu 1 Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA Full list of author information is available at the end of the article Yu et al. Cell Biosci (2021) 11:94 Page 2 of 12 intramedullary, juxtacortical, and extraskeletal OSs, and each of these OS types can be further divided into several subtypes [6]. Intramedullary OS is the most common and the fastest growing type, which accounts for nearly 80% of all OSs. Intramedullary OS develops in the medullary cavity of a long bone, such as femur and tibia. Further- more, there are a number of subtypes of intramedullary OS according to the tumor cell types. The common sub- types include osteoblastic, chondroblastic, fibroblastic, small-cell, and epithelioid OSs. Juxtacortical OS is the second most common type, and they account for 10 to 15% of all OSs. This type of OS develops on the outer sur- face of the bone, or periosteum that is the dense layer of connective tissue covering the bone. Extraskeletal OS is rare and grows slowly, accounting for fewer than 5% of all OSs. Extraskeletal OS does not touch the bone and often arises from soft tissues that have experienced prior radia- tion therapy (Fig. 1a). patients over the age of 60 are the secondary OSs. Unlike OSs in children and young patients, OSs in elder patients more commonly develop at axial locations and in tissue areas that have previously received irradiation or have existing bone abnormalities [11]. OS patients older than 60 years are associated with a higher risk of metastatic disease [8]. Elder men are also associated with higher risks to develop OS than elder women do. In contrast to the OS disparity in black children, OS is more fre- quently observed in white elder adults compared with black and other elder adults [10]. The incidences of OS among 24-year-old and younger individuals are generally consistent in different countries in the world, with most cases diagnosed during puberty. However, the incidences of OS among elder men are higher in some countries including United Kingdom, Australia, and Canada com- pared with other countries [12]. Most OSs occur sporadically, and the exact causes for OS development are still not fully understood. Especially, the vast majority of OS cases in adolescents and young adults are sporadic with no known familial genetic or environmental causes [13–15]. However, it has been noticed that most OS cases occur in patients with cer- tain rare inherited types of cancers or diseases such as retinoblastoma, Li–Fraumeni syndrome, and Roth- mund–Thomson syndrome, which involve chromosomal abnormalities of the tumor-suppressors RB1 and TP53, as well as the DNA helicase REQL4 genes, respectively [6, 11, 16]. Individuals carrying germline RB1 mutations have approximately 1000-fold increased risk to develop OS [17]. Abnormalities in the CDKN2A gene, which codes for p16INK4a, a CDK4 inhibitor, and p14ARF, a p53 stabilizer, also increase the risk of OS development [6]. OS occurs more commonly at the regions of bone growth, which is presumably attributed to the genomic mutations acquired from the active cell proliferation. Most OSs occur in children and young adults at ages from 10 to 30 years old, with the peak incidence in the second decade of life; however, people at any ages can develop OS [7]. OS is the third most common cancer in adolescence, accounting for approximately 3% of all ado- lescent cancers, with only lymphoma and brain tumor being more prevalent [1]. Adolescent OS usually devel- ops in a region where the bone grows rapidly, such as the end region of the long leg or arm bones [8]. In some cases, OS occurs in the humerus, ulna, radius, fibula or pelvis (Fig. 1b) [9]. OS occurs slightly more common in boys versus girls, which may be related to the taller body height correlated with faster bone growth in boys on average. However, girls tend to develop OS slightly ear- lier, which may be related to their earlier growth spurt on average. The incidence of OS is higher in black popula- tion compared with white and other populations [10]. In the United States, more than 50% of all OS arising in b 42% Femur 19% Tibia 8% Jaw 13% Other rcomas b 42% Femur 19% Tibia 8% Jaw 13% Other Osteosarcoma (OS) Intramedullary OS (～～80%) Juxtacortical OS (～～10-15%) Extraskeletal OS (～～5%) Osteoblastic OS Condroblastic OS Fibroblastic OS Small cell OS Epithelioid OS Other a Fig. 1 The types and subtypes (a) as well as the skeletal distribution (b) of osteosarcomas b a Yu et al. Cell Biosci (2021) 11:94 Page 3 of 12 People receiving radiation therapies for treating other types of cancers may have a higher risk to develop OS later from the treated tissue sites. Animal OS‑derived cell lines Along with the rapid development of cancer immuno- therapy, cancer cell lines isolated from spontaneously developed animal tumors have become important syn- geneic models for studying immune suppression and activation, as well as interactions between immune and cancer cells in the same strain animals with identical genetic background (Table 1). The first batch of success- fully established metastatic OS cell lines including K7, K8, K12, K14 and K37 were derived from a spontaneous OS in the distal femur of an 895-day-old female BALB/c mouse [34, 36–40]. These lines were metastatic in vivo and have been used for years to study the process of OS metastasis [41]. The Dunn cell line is another murine OS cell line derived from a spontaneous OS in the tail of a C3H/HeN mouse. This cell line is metastatic in vivo, and its xenograft tumors commonly metastasize to lung and liver in mice [42]. Multiple sublines derived from Dunn cell line have been used as OS angiogenesis and metas- tasis models in screening new compounds and testing candidate drugs [43–46]. The UMR 106-01 cell line was developed from a 32P-induced OS tumor in a Sprague- Dawley rat [32, 47]. This cell line has been well adopted to OS research due to its phenotypical similarities to human OS cells and rapid formation of pulmonary metastasis [48–51]. Besides murine, a number of cell lines have been derived from spontaneous canine OSs. Among these canine OS cell lines, the D-17 cell line was isolated from the lung metastasis of an 11-year-old female poodle. D-17 cells have been widely used in finding therapeutics for bone cancers in dogs [52, 53] (Table 1). Cellular and animal models for studying OS Canine OS models Spontaneous OSs are more common in large dogs com- pared with humans, making dog an attractive model to study this disease [20]. Canine OS is similar to that of human OS in terms of biological features and clini- cal symptoms [21, 22]. It is estimated that over 10,000 cases of canine OS occur annually in the United States. The major difference between canine OS and human OS is that canine OS is a disease of large breed elder dogs (6–12 years of age), which is considered as a limitation to use canine OS models to study human OS. The median disease-free intervals are 4 months after single surgery treatment, and 13 months after combined treatment of surgery and chemotherapy [23–25]. Many genomic alterations involved in human OS pathogenesis are also detected in canine OSs, such as the loss-of-function genetic alterations of the TP53 [26–28], RB [29] and PTEN [30] tumor suppressor genes in both human and canine OSs. Indeed, people treated with radiotherapies at younger ages or received high irra- diation doses have increased risks to develop OS [11]. In addition, people with certain non-cancerous bone diseases such as Paget’s disease and hereditary multiple osteochondromas also have increased risks to develop OS. Specifically, about 1% of Paget’s disease patients actually develop OS [11]. [31, 32]. Murine OS models have been induced by expos- ing animals to radioactive substances such as radium, thorium and roentgen [33]. Despite their high inci- dences, these models probably represent secondary OSs developed in human patients who have received previous irradiation treatment, which might not share molecular mechanisms with primary OSs. Most of the chemically induced mouse OS models have been developed by injecting different chemical carcinogens directly into bones [33]. For example, rats treated with P32-orthophosphate have been shown to develop OS tumors that histologically resemble human OS [31, 34]. However, the carcinogen-induced murine model is more representative of a therapy-induced disease, while most human OSs are sporadic [35]. About 20% of OS have disseminated to the lung, brain or other organs at the time of diagnosis [18]. Chemo- therapy and surgery are the most common treatments for most OS patients [1, 19]. Chemotherapies given before and/or after the surgery significantly reduce the risk of recurrence. The most commonly used chemotherapy drugs include methotrexate, doxorubicin and cisplatin, and two or more of these drugs are usually administrated in combination [19]. The 5-year survival rate of all-stage OSs is about 60%. However, if metastasis has happened at the time of diagnosis, the 5-year survival rate drops significantly to only 27%, which has not been improved significantly over the last four decades [16]. Recurrent or metastatic OSs are usually resistant to currently available standard treatments. Therefore, it is necessary to under- stand the detailed mechanisms responsible for OS devel- opment, progression and metastasis in order to identify novel therapeutic targets for treating OS. Human OS‑derived cell linesi U2OS, the first human OS cell line, was established in the year of 1964, and has been used extensively in many in vitro studies. One limitation of this cell line is that it does not satisfy researchers’ needs for an in vivo Radiation or chemical carcinogen induced OS models Radiation or chemical carcinogen induced OS models Historically, rodent OS models began with the exposure of rats or mice to chemical and radioactive carcinogens Yu et al. Cell Biosci (2021) 11:94 Page 4 of 12 Table 1 The origin, morphology, genetic mutation, tumorigenic capability, and metastatic potential of OS cell lines m/m, mutant/mutant homozygous; w/w, wild type/wild type normal alleles; m/w, mutant/wild type heterozygous; NA, not available; E, epithelial; F, fibroblast; O, osteoblast; Meta, metastasis Cell line Origin Morphology Gene mutation Days to form 1.5–2 cm3 tumor Meta rate (%) Refs. Tumor suppressor Oncogene CDKN2A TP53 RB1 PTEN BRAF KRAS HOS OS of a 13-y-o girl E m/m m/m w/w w/w w/w w/w No tumor in 8 weeks 0% [58, 64] 143B HOS cells, transformed by KRAS Mixed E/F m/m m/m w/w w/w w/w m/w 27 100% [64, 69] MNNG HOS cells, induced by MNNG Mixed E/F m/m m/m w/w w/w w/w w/w 32 100% [64, 66, 70] SaOS-2 OS of a 11-y-o girl E w/w m/m m/m w/w w/w w/w 95 100% [62, 63] SaOS-LM7 Lung Meta from SaOS-2 cells in mouse tibia E w/w m/m m/m w/w w/w w/w 65 100% [71] MG-63 Fibroblastic OS of a 14-y-o boy F m/m m/m w/w w/w w/w w/w 170 0% [59] U-2 OS Sarcoma in tibia of a 15-y-o girl E w/w w/w w/w w/w w/w w/w No tumor in 150 days 0% [60, 61, 72] K7 (K8, …, K12) Spontaneous OS in BALB/c mouse O NA NA NA NA NA NA NA 33.3% (K12) [37, 38, 45] K7M2 Lung meta From K7 cells in mouse tibia O NA NA NA NA NA NA 30 days to reach 0.8 cm3 93.3% [36, 37, 39, 40, 45] Dunn Spontaneous OS from C3H/HeN mouse tail E NA NA NA NA NA NA 25 100% [42, 45] UMR 106-01 32P-induced tumor in rat E NA NA NA NA NA NA 25 days to reach 0.15 cm3 100% [32, 47] D-17 Lung Meta of OS in a 11-y-o female poodle E NA NA NA NA NA NA No tumor in 79 days NA [53] Yu et al. Cell Biosci (2021) 11:94 Page 5 of 12 metastatic model [54–61]. Unfortunately, this was also the case for other several subsequently established human OS cell lines such as HOS and SaOS2 cell lines [58, 62, 63]. Genetically engineered OS mouse models Genetically engineered OS mouse models Cell culture models may impose widespread genetic changes and loss of phenotypic heterogeneity that diverge from the characteristics of the original OSs. Tak- ing SaOS2 as an example, OS cells maintained in culture demonstrate significant changes in phenotype over time [73]. In general, higher passage cells exhibit higher pro- liferation rates and lower alkaline phosphatase activity, while mineralization is more pronounced in late passage cells. Gene expression profiles may also change in culture over time. Genetically engineered mouse (GEM) models may help to provide spontaneously developed OS models with natural tumor environment for studying OS initia- tion, growth and metastasis.hi Radiation or chemical carcinogen induced OS models In 2005, the HOS cells were treated with carcinogen or virus-mediated oncogene expression to induce genetic alterations, and the first metastatic human OS cell line 143B, along with several other derivative cell lines were established from these treated cells [58, 64–71]. Thereafter, many metastatic cell lines have been established in vitro, which have formed the basis for stud- ying the cellular and molecular processes of OS (Table 1). A recent study characterized a set of 19 OS cell lines by profiling their gene expression and epigenetic patterns, and by comparing their differentiation, growth, invasion, and migration capacities in nude mice [72]. These valua- ble data should facilitate investigators to select appropri- ate OS cell lines for their researches. osteogenic cell lineages to model human OSs. Deletion of both Tp53 and Rb genes by Osterix-Cre leads to OS development with high penetrance [80]. Mice with Prx-1- Cre mediated deletion of both Tp53 and Rb genes induces OS development, and also generated poorly differenti- ated soft tissue sarcomas [81]. Mice with Osterix-Cre activated expression of a transgenic shRNA that targets p53 mRNA also develop osteoblastic OS at 100% pene- trance. Although this model exhibits a longer latency to tumor onset, these OSs often develop in long bones and are highly metastatic to lung and liver. More importantly, this model does not develop any non-OS tumors [82]. In addition to c-fos and p53, other proteins such as TWIST1 [83], p14ARF [84], p16INK4a [85], PRKAR1A [86], and p21CIP [87] have been implicated in OS patho- genesis based on findings from human OS samples, and their involvements in OS development have also been demonstrated in GEM models. Their alterations appear to complement the defects in the p53 and Rb pathways. Although these models provide further insights into OS genetics and biology, the long latency combined with low penetrance makes these models less practical. OS cell‑derived xenograft models Human or mouse OS cells are routinely inoculated into immunocompromised mice to grow xenografts and allo- grafts as OS models. The injected cells usually develop solid tumors within days or weeks. The advantages of these models include quick onset, affordable cost, ease of handling and maintenance, and high reproducibility. Although OS cells are ectopically inoculated under the skin in some studies, cell grafts that grow orthotopically in or near bones are considered as more relevant preclin- ical OS models. These OS cell-derived xenograft models have been very useful in identification of factors that are involved in OS invasion and drug candidates that inhibit OS growth [36, 37, 41, 88]. An obvious limitation of these xenograft models is that they do not provide information about the initiation and etiology of OS since it uses fully immortalized OS cells. The first GEM model of OS is the H2K-fos-tg mouse model, where c-fos is overexpressed in osteoblasts to induce OS development. The tumors developed in this model display similar histopathology to human osteo- blastic OS, but these tumor cells do not produce distant metastasis that frequently occurs in human OSs [74]. Many murine OS models have been developed to reca- pitulate p53 and RB mutations in hereditary and sporadic human OSs [75]. Germ-line deletion of p53 results in an OS incidence of 4% in homozygous p53 null mice [76] and 25% in heterozygous p53 mice [77], indicating the importance of p53 loss in OS development. The higher OS incidence in heterozygous versus homozygous p53 knockout mice may be due to the development of other types of cancer that results in early death of the homozy- gous knockout animals. However, homozygous Rb knockout mice are lethal before birth, and heterozygous Rb knockout mice do not develop OS [78, 79]. Patient‑derived xenograft (PDX) OS models Patient‑derived xenograft (PDX) OS models To establish a PDX line, small pieces of fresh tissue from either an incisional (open) biopsy or a percutaneous (needle) biopsy of an OS tumor in a patient are trans- planted into multiple immune-defective mice to grow xenograft tumors [89]. In PDX models, the OS tumor cells are never cultured in vitro and always maintained in an OS-like tissue environment, and thus, PDX models are considered as a much more clinically relevant model to represent both the general features and heterogeneity of human OSs [90, 91]. The PDX models also allow the study of early-stage progression of OS metastasis in vivo. The application of conditional gene manipulation and tissue specific Cre expression in mice have greatly enhanced the ability to induce OS from mesenchymal Yu et al. Cell Biosci (2021) 11:94 Page 6 of 12 The PDX models have been broadly used for screening and testing drugs for developing new therapies [92–94]. However, like any other models, the PDX models are not perfect. Some initial generations of PDXs may take a long time to form tumors, suggesting that these OS cells still experience senescence, immortalization, and/or growth selection processes before a small population of tumor cells grows to form a tumor. Additionally, along with the tumor growth, most, if not all, of the stromal cells such as fibroblasts, immune cells, vascular cells and fat cells in the tumor environment are replaced with mouse cells, which means that the human OS tumor cells still grow in a mouse tissue and cellular environment. can have various degrees of EMT or MET statuses, rang- ing from full epithelial to full mesenchymal states (Fig. 2) [95]. Most sarcoma cells including OS cells exhibit an epithelial or a hybrid mesenchymal and epithelial phe- notype. Since sarcoma cells are inherently locked in a mesenchymal state, they unlikely reprogram to a full epithelial state. However, depending on sarcoma’s histio- types, they can reprogram their degrees of epithelial and mesenchymal states during their growth, progression and metastasis. Indeed, although the features of epithelial and mesenchymal plasticity are variable across OSs with dif- ferent histiotypes, EMT and MET events are frequently observed and regulated by many molecular players [98]. EMT‑promoting transcription factors (EMT‑TFs) in OS and other sarcomas p g Epithelial-to-mesenchymal transition (EMT) and its reverse process, mesenchymal-to-epithelial transition (MET), are required for adapting cell plasticity in many physiological and pathophysiological processes, such as embryonic development, wound healing, fibrosis and cancer metastasis [95]. The cancer cells with an epithelial origin such as breast and prostate adenocarcinoma cells can undergo EMT to acquire mesenchymal gene mark- ers and morphologies. Cancer cells at an EMT state are highly capable to escape from epithelial tumor cell clus- ters, invade into the stromal tissue, and disseminate to distant organs [96, 97]. Once getting in a distant organ, a MET process is thought to help the disseminated can- cer cells to adapt the new tissue environment for prolif- eration and establishment of metastasis [96]. Cancer cells EMT has been extensively studied in carcinomas in which EMT properties are associated with drug resist- ance, invasion and metastasis [96]. Sarcoma has a mes- enchymal origin, and its mesenchymal phenotype is maintained by the functions of EMT-TFs, including TWIST1, SNAIL, SLUG, ZEB1 and ZEB2, and associated with more aggressive behaviors. Depending on different patient cohorts, variable percentages (32–56%) of human OSs have TWIST1 expression. The TWIST1-positve OS cells are associated with metastatic phase III OS tissues and also with poor clinical outcomes [99, 100]. SNAIL is widely expressed in OSs [101]. Knockdown of SNAIL in SaOS2 cells increases E-cadherin expression to promote MET, which is accompanied with decreased cell migra- tory and invasive properties. Conversely, overexpression N N E N E N E N N N N E E N E MET EMT Proliferation Invasion Stemness Epithelial markers Mesenchymal markers Epithelial (E) E/M Mesenchymal (M) ++ + - + ++ +++ + ++ - + ++ ++ + ++ ++ Cell-cell adhesion ++ + - Motility + ++ ++ Fig. 2 The different cellular features associated with the different plasticity states of osteosarcoma (OS) cells. In this model, OS cells can maintain different states of hybrid epithelial/mesenchymal phenotypes characteristic of different expression levels of epithelial and mesenchymal markers. These OS cells can undergo either more epithelial or more mesenchymal transition states, which are correlated with different cellular features associated with the aggressiveness of OS. EMT, epithelial-to-mesenchymal transition; MET, mesenchymal-to-epithelial transition; V, vimentin; E, E-cadherin; N, nucleus. +++, strong or high; ++, moderate; +, weak; −, negative N N N MT Mesenchymal (M) - +++ - ++ ++ - ++ Fig. The signaling pathways and EMT‑TFs that regulate MET The signaling pathways and EMT‑TFs that regulate MET Several signaling pathways can program a MET status of OS cells through regulating the transcriptional activities of EMT-TFs. These cross-talk regulatory networks are depicted in Fig. 3. Specifically, blocking Wnt/LDL recep- tor related protein 5 (LRP5) signaling by a soluble nega- tive dominant form of LRP5 mutant in OS cells markedly upregulates the expression of E-cadherin, an epithelial marker, and downregulates the expression of N-cad- herin, a mesenchymal marker. Inhibition of the Wnt/ LRP5 signaling also downregulates the activity of EMT- TFs such as TWIST1 and SLUG [113]. Synovial sarcoma translocated-synovial sarcoma X1 and 2 (SYT-SSX1/2) interact with SNAIL and SLUG, respectively, to dimin- ish their transcriptional repression activities on E-cad- herin expression, resulting in an increase in E-cadherin expression and an acquisition of epithelial characteristics in synovial sarcoma cells [112]. In OS cells, the autocrine motility factor (AMF), also known as phosphoglucose isomerase (PGI), enhances SNAIL activity. Accordingly, silencing the expression of AMF/PGI can reduce SNAIL activity, which induces terminal differentiation of these OS cells into mature osteoblasts, resulting in suppression of the growth and pulmonary metastasis of these OS cell- derived xenografts in nude mice [114]. Ovo like zinc fin- ger 2 (OVOL2) represses ZEB1 expression by binding to the ZEB1 promoter, so high OVOL2 expression is associ- ated with low ZEB1 expression in human OS. In agree- ment with this finding, overexpression of OVOL2 in OS cells can promote MET and suppress cell migration and invasion [115]. In addition, miRNAs can directly regu- late E-cadherin expression to induce MET or indirectly regulate E-cadherin expression through targeting EMT- TFs. For example, miR-30 and miR-9 can target TWIST1, SNAIL and ZEB1 mRNAs, and miR-23b, miR-29c, miR- 194 and miR-200 can downregulate TWIST1 and ZEB1 mRNAs, resulting in upregulation of E-cadherin expres- sion [116, 117]. It is interesting to notice that, expression EMT‑promoting transcription factors (EMT‑TFs) in OS and other sarcomas 2 The different cellular features associated with the different plasticity states of osteosarcoma (OS) cells. In this model, OS cells can maintain different states of hybrid epithelial/mesenchymal phenotypes characteristic of different expression levels of epithelial and mesenchymal markers. These OS cells can undergo either more epithelial or more mesenchymal transition states, which are correlated with different cellular features associated with the aggressiveness of OS. EMT, epithelial-to-mesenchymal transition; MET, mesenchymal-to-epithelial transition; V, vimentin; E, E-cadherin; N, nucleus. +++, strong or high; ++, moderate; +, weak; −, negative Yu et al. Cell Biosci (2021) 11:94 Page 7 of 12 of SNAIL in SaOS2 cells suppresses E-cadherin expres- sion to promote EMT-like process, which promotes invasion and metastasis of these cells [102]. The expres- sion of SLUG is associated with high-grade cranial bone OSs with high metastatic potentials [103]. Knockdown of SLUG in canine D-17 and human SaOS2 cells signifi- cantly decreases the migration and invasion capabilities of these OS cells by remodeling their actin cytoskeleton organization and disrupting their cellular protrusions. Knockdown of SLUG also inhibits the growth of these cell-derived xenografts in an in vivo chick chorioallantoic membrane (CAM) assay model. Ectopic expression of SLUG in these OS cells increases the expression level of WNT5A, decreases the expression level of the adhesion molecule osteoblast cadherin (OB-Cad), and increases cell motility by promoting the formation of actin-rich cellular protrusions [104]. The expression levels of ZEB1 protein in human sarcoma tissues are also positively cor- related with lung metastasis, which is consistent with the finding showing that ZEB1 knockdown in MG-63 cells significantly inhibits cell invasive capability [105]. The mixed canine benign mammary tumors are composed of epithelial cells and cartilage or bone tissue, which is a species-specific type. The malignant canine mammary tumors include carcinomas, fibrosarcomas and OSs. In comparison with canine mammary epithelial cells and carcinomas, fibrosarcomas express high and OSs express even higher levels of a panel of homeobox genes includ- ing ZEB2. ZEB2 is an EMT-TF important for craniofacial bone formation [106]. Collectively, these studies suggest that the EMT-TFs play important roles in maintaining the mesenchymal status of sarcomas. Understanding the molecular mechanisms underlying these EMT-TFs regu- lated OS cell growth and metastasis may provide new opportunities to identify potential molecular targets for treating OSs. of SNAIL in SaOS2 cells suppresses E-cadherin expres- sion to promote EMT-like process, which promotes invasion and metastasis of these cells [102]. EMT‑promoting transcription factors (EMT‑TFs) in OS and other sarcomas The expres- sion of SLUG is associated with high-grade cranial bone OSs with high metastatic potentials [103]. Knockdown of SLUG in canine D-17 and human SaOS2 cells signifi- cantly decreases the migration and invasion capabilities of these OS cells by remodeling their actin cytoskeleton organization and disrupting their cellular protrusions. Knockdown of SLUG also inhibits the growth of these cell-derived xenografts in an in vivo chick chorioallantoic membrane (CAM) assay model. Ectopic expression of SLUG in these OS cells increases the expression level of WNT5A, decreases the expression level of the adhesion molecule osteoblast cadherin (OB-Cad), and increases cell motility by promoting the formation of actin-rich cellular protrusions [104]. The expression levels of ZEB1 protein in human sarcoma tissues are also positively cor- related with lung metastasis, which is consistent with the finding showing that ZEB1 knockdown in MG-63 cells significantly inhibits cell invasive capability [105]. The mixed canine benign mammary tumors are composed of epithelial cells and cartilage or bone tissue, which is a species-specific type. The malignant canine mammary tumors include carcinomas, fibrosarcomas and OSs. In comparison with canine mammary epithelial cells and carcinomas, fibrosarcomas express high and OSs express even higher levels of a panel of homeobox genes includ- ing ZEB2. ZEB2 is an EMT-TF important for craniofacial bone formation [106]. Collectively, these studies suggest that the EMT-TFs play important roles in maintaining the mesenchymal status of sarcomas. Understanding the molecular mechanisms underlying these EMT-TFs regu- lated OS cell growth and metastasis may provide new opportunities to identify potential molecular targets for treating OSs. expression of tight junction proteins such as claudin-1 and ZO-1 in over 50% of ES/PNET samples, suggesting partial epithelial differentiation in this kind of cancer [109]. It is also reported that patients with leiomyosarco- mas that express high epithelial signature genes includ- ing E-cadherin also have a better prognosis [110, 111]. In OSs, E-cadherin expression levels have been found to be inversely correlated with metastasis potential but posi- tively correlated with good prognosis [99]. However, it should be noted that the process of MET in sarcoma is characterized by increased expression of epithelial-like markers such as E-cadherin, whereas the typical mesen- chymal markers including vimentin remain abundantly expressed in the sarcoma cells [111, 112]. MET in OS and other sarcomas Since sarcomas have a mesenchymal cellular origin, more studies have been carried to understand the role of MET in sarcoma progression. Patients with more epithelial-like carcinomas tend to have better clinical outcomes com- pared with patients with more mesenchymal-like carci- nomas, and a similar trend is also the case for patients with sarcomas. E-cadherin is the first epithelial marker detected in bone and soft tissue sarcomas [107]. A meta- analysis of 812 bone and soft tissue sarcoma tumors demonstrates that low E-cadherin expression is associ- ated with poor five-year overall survival [108]. Ewing sar- coma/primitive neuroectodermal tumor (ES/PNET) cells frequently express epithelial markers such as cytokerat- ins, claudin-1 and ZO-1, and exhibit a partial epithelial differentiation state. This study showed evidence for Yu et al. Cell Biosci (2021) 11:94 Page 8 of 12 SNAIL SLUG TWIST1 ZEB1 SYT-SSX1/2 LRP5 AMF/PGI VDR TIMP1 OVOL2 miR-30 miR-9 miR-23b miR-29c miR-194 miR-30 miR-9 miR-200 E-cadherin MET Fig. 3 The molecular regulatory mechanisms for MET in OS cells. The EMT-inducing transcription factors including SNAIL, SLUG, TWIST1 and ZEB1 are expressed in OS cells which directly or indirectly repress the expression of epithelial genes such as E cadherin to maintain mesenchymal SNAIL SLUG TWIST1 ZEB1 SYT-SSX1/2 LRP5 AMF/PGI VDR TIMP1 OVOL2 miR-30 miR-9 miR-23b miR-29c miR-194 miR-30 miR-9 miR-200 E-cadherin MET Fig. 3 The molecular regulatory mechanisms for MET in OS cells. The EMT-inducing transcription factors including SNAIL, SLUG, TWIST1 and ZEB1 are expressed in OS cells, which directly or indirectly repress the expression of epithelial genes such as E-cadherin to maintain mesenchymal cell features. MET is initiated by inhibiting EMT-TFs through activating upstream signaling pathways such as SYT-SSX1/2, OVOL2 and miRNAs or suppressing AMF/PGI or LRP5 in OS cells. Please refer to the text for related references. VDR vitamin D receptor, TIMP1 TIMP metallopeptidase inhibitor 1 miR-30 miR-9 AMF/PGI MET E-cadherin Fig. 3 The molecular regulatory mechanisms for MET in OS cells. The EMT-inducing transcription factors including SNAIL, SLUG, TWIST1 and ZEB1 are expressed in OS cells, which directly or indirectly repress the expression of epithelial genes such as E-cadherin to maintain mesenchymal cell features. MET is initiated by inhibiting EMT-TFs through activating upstream signaling pathways such as SYT-SSX1/2, OVOL2 and miRNAs or suppressing AMF/PGI or LRP5 in OS cells. Please refer to the text for related references. MET in OS and other sarcomas VDR vitamin D receptor, TIMP1 TIMP metallopeptidase inhibitor 1 EMT-TFs may also serve as potential therapeutic tar- gets in sarcomas. It has been reported that TWIST1 is one of the 45 chemoresistant-signature genes that can predict OS patients’ response to neoadjuvant chemother- apy at the time of diagnosis. Knockdown of TWIST1 in multiple OS cell lines including HOS, SJSA-1 and 143B cells can largely overcome the chemoresistance of these cancer cells [121]. However, another study showed that TWIST1 can increase the chemosensitivity of SaOS2 and MG-63 OS cells to cisplatin treatment by downregulat- ing endothelin-1 (ET-1) [122], because high ET-1 expres- sion increases cell invasion and survival against cisplatin treatment in OS cells [123]. These different results from different studies might be due to the different func- tions of TWIST1-regulated target genes. Therefore, it may depend on specific OS cell context to determine whether TWIST1 can serve as a drug target. It has also demonstrated that OS cells that survived after cisplatin treatment at a sublethal dose exhibit a more mesenchy- mal phenotype and an elevated capacity to metasta- size. Under these circumstances, inhibition of SNAIL can promote cisplatin sensitivity and prevent cisplatin treatment-induced EMT-like process, which results in diminished OS cell growth and survival [124]. Therefore, targeting these EMT-TFs in OS cells may help to induce MET and improve OS cell response to chemotherapy. of one mesenchymal factor is often sufficient to induce EMT in epithelial-derived carcinomas, while expression of two epithelial factors such as GRHL2 and miR-200 are required to drive MET in human rhabdomyosarcoma cells. This may suggest that the expression of epithelial genes in mesenchymal cells requires both transcriptional de-repression (e.g. via miR-200s) and transcriptional acti- vation (e.g. via GRHL2) events to happen [118, 119]. Availability of data and materials Not applicable. Availability of data and materials Not applicable. Author details It becomes obvious that new targets still need to be identified for developing new therapeutic strategies and drugs. Further understanding of the cell plasticity in OS progression could offer new opportunities to address these issues. EMT-TFs, especially TWIST1 and SLUG, play important roles in bone development and remod- eling. These EMT-TFs regulate EMT and MET plastic- ity of OS cells, which is not identical to their regulations in solid tumors of epithelial origin [98]. Further inves- tigation and deeper understanding of the EMT/MET- regulatory machineries in OS cells may help to identify druggable molecular targets. Furthermore, given the complexity of EMT/MET-like regulatory networks and the ability of cancer cells to adapt to stress conditions, targeting one protein or pathway may not be sufficient to completely impede EMT-related process or initiate MET. The future druggable targets that can be identified from the EMT/MET-like regulatory networks for treating 1 Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA. 2 Department of Pediatrics, Texas Children’s Cancer and Hematology Center, and The Faris D. Virani Ewing Sarcoma Center, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA. Received: 4 March 2021 Accepted: 30 April 2021 Clinical implications and therapeutic opportunities of the EMT/MET plasticity of OS cellsh The growing body of data on the regulation of sarcoma cell EMT/MET plasticity may offer clinical implications and therapeutic opportunities to patients. Specifically, the MET phenotype with epithelial marker expression in sarcomas may serve as prognostic markers. Withaf- erin-A (WFA), a naturally derived bioactive compound, is a vimentin inhibitor, which could be a promising drug against vimentin-expressing sarcoma cells. WFA treat- ment causes vimentin cleavage and induces sarcoma cell apoptosis. WFA also significantly inhibits growth, local recurrence and metastasis of the soft tissue sarcoma cell- derived xenografts in vivo [120]. This finding suggests that vimentin may be a good target for inhibiting soft tis- sue sarcomas. Yu et al. Cell Biosci (2021) 11:94 Yu et al. Cell Biosci (2021) 11:94 Page 9 of 12 Conclusion remarks metastasis of OS cells may be used in combination with the current surgery and chemotherapy treatments to achieve better clinical outcomes. Metastasis is the major cause of cancer-related death. When compared with numerous metastasis-related researches carried out for other cancer types such as breast and prostate cancers, much fewer studies of OS metastasis have been conducted so far. OS patients treated in recent years basically still receive essentially unchanged treatments applied in 1970s [16]. These may be due to the absence of consistent genetic mutations, and the relatively low incidence and high heterogeneity of the disease. Furthermore, our limited understanding of the biology about what drives OS cell dissemination from the primary bony sites and their subsequent proliferation at a second tissue environment such as the lung may have hindered our ability to develop new therapies for treating metastatic OSs. To impact the lives of patients suffering from metastatic OSs, it will be necessary to deepen our fundamental knowledge about OS metastasis and its spe- cific vulnerabilities at cellular and molecular levels. The cell signaling pathways implicated in OS biology through genetic and other preclinical studies mainly include PI3K/mTOR [125], TGFβ [126], RANKL/NF-κB [127], and IGF [128]. Unfortunately, clinical studies evaluating the reagents that target these pathways have yielded dis- appointing results. Recently, although HER2 expression has been detected in certain OSs and tested as a thera- peutic target in OS, targeting HER2 with trastuzumab, an FDA approved antibody drug for breast and gastric cancers with HER2 overexpression, is still not very effec- tive to inhibit OS growth [129]. However, immunother- apy using HER2 chimeric antigen receptor (CAR) T-cells may be developed into a promising therapeutic approach for treating HER2-positive OSs [130]. Of note, the role of HER2 in the regulation of EMT/MET plasticity in OS cells is currently unknown. Authors’ contributions JX and XY performed and conceived the review. XY wrote the manuscript and drew all the figures. JX and JY revised the manuscript and figures. All authors read and approved the final manuscript. Abbreviations AMF: Autocrine motility factor; CAM: Chick chorioallantoic membrane; EMT: Epithelial-to-mesenchymal transition; EMT-TF: Epithelial-to-mesenchymal tran- sition transcription factor; ES/PNET: Ewing sarcoma/primitive neuroectoder- mal tumor; ET-1: Downregulating endothelin-1; GEM: Genetically engineered mouse; LRP5: LDL receptor related protein 5; MET: Mesenchymal-to-epithelial transition; Meta: Metastasis; OB-cad: Adhesion molecule osteoblast cadherin; OS: Osteosarcoma; OVOL2: Ovo like zinc finger 2; PDX: Patient-derived xenograft; PGI: Phosphoglucose isomerase; SYT-SSX1/2: Synovial sarcoma translocated-synovial sarcoma X 1/2; TIMP1: TIMP metallopeptidase inhibitor 1; VDR: Vitamin D receptor; WFA: Withaferin-A. Competing interests All authors do not have any conflict of interests to disclose. All authors do not have any conflict of interests to disclose. 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https://openalex.org/W3131140898	OpenAlex	Open Science	CC-By	2,021	Comparative study for connecting new flare capacity to existing flare Systems	Mohamed Shamandy Fouad Et al.	English	Spoken	5,911	13,872	PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 Keywords Flaring system; depressurization; emergency shutdown; conventional blowdown; sequential blowdown; maximizing relieving capacity. Article Received: 18 October 2020, Revised: 3 November 2020, Accepted: 24 December 2020 Flare is the last line of defense in the safe emergency release system in the petroleum and chemical plants. The flare provides a means of safe disposal of the vapor streams from its facilities, by burning them under controlled conditions. Such that adjacent equipment or personnel are not exposed to hazards, and at the same time obeying the environmental regulation of pollution control and public relations requirements. The chemical process used for flaring is a high temperature oxidation reaction to burn combustible components, mostly hydrocarbons, or waste gases from industrial operations [3]. Comparative study for connecting new flare capacity to existing flare Systems Mohamed Shamandy Fouada, Walaa Mahmoud Shehatab , Fatma Khalifa Gadb, Nabil Abdelsadek Abdelaleem c a KHALDA Petroleum Company, Maadi, Egypt b Suez University, Faculty of petroleum and Mining Engineering, Department of petroleum Refining and Petrochemical Engineering. c GENERAL Petroleum Company, Nasr City, Egypt ABSTRACT Process facility operations are equipped with flare system to dispose flammable, toxic, or corrosive vapors to an environmentally acceptable gas for release to the atmosphere from both normal operational venting and relief during abnormal conditions. For safe incineration and radiation considerations flares are located at a remote point from the plant. Flare system when designed is to be limited for a certain capacity for a relieved gas; in addition, it may be designed in excess for future process facilities which may be further added to an existing one. KHALDA Petroleum Company is an owner company of oil and gas plants in Egypt. The company started a remote facility in 2005 named as "QASR start of line" which equipped with a limited flaring system capacity of 416,800 kg/hr for each flare related to phases PH-1 and PH-2. After 10 years of operation the wells depletion occurred which affected the production capacity and hence the company decided to maintain the productivity. A new compression project is designed to improve recovery as the reservoir production rate and pressure decline. The new compression project facilities vent and disposal need a flare system in case of the emergency. This paper discusses two different scenarios available for connecting the new compression project facilities vent and disposal system either to the existing flare systems or to another destination, which better, safer and more economic Introduction In most process facility operations gas and vapor have to be disposed of safely, quickly, and without environmental impact [1]. When the venting gas or vapor cannot be converted to useful energy it is routed to a remote point for safe incineration, which is called faring. Flares are the most economical and customary means of disposing of excess light combustible gases in the process industries. The primary function of a flare is to convert flammable, toxic, or corrosive vapors to an environmentally acceptable gas. These acceptable gases are released to the atmosphere from both normal operational venting and relief during abnormal conditions [1, 2]. 5795 In combustion, the gaseous hydrocarbon (natural gas, propane, ethylene, propylene, butadiene, www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 line (SOL) Qasr Plant. The Qasr gas and condensate free flows from the wellheads through the Qasr Phase I and Phase II facilities and export pipelines to the SHAMS manifold and Salam gas plant under reservoir pressure until year 2015. As the reservoir pressure declines the peak gas rate of 800 mmscfd are no longer be achievable. The Qasr Compression Project is designed to improve recovery as the reservoir production rate and pressure decline. butane and etc.) reacts with atmospheric oxygen to form carbon dioxide (Co2) and water. Several byproducts formed will be carbon monoxide, hydrogen and others dependent upon what is being burned. Efficiency of hydrocarbon conversion is generally over 98% [4-8]. The flare system must be correctly sized to handle the material vented during depressuring, especially peak flow. During the blowdown operation, a large amount of material must be disposed of simultaneously. It is necessary to ensure that there is sufficient flare discharge capacity to do this without violating hydraulic constraints in the flare piping. This causes overpressures or excessive vibrations or exceeding radiation limits from the flare tip. It is also necessary to accurately determine what size of restriction orifices or other flow-controlling devices and flare connections are required for depressuring each section of the plant [9-11]. There are two existing independent HP flare systems, one for Phase I facilities and the other for Phase II facilities. Phase I HP flare system, comprising a duplex stainless steel collection system, carbon steel KO drum, carbon steel riser and stack on which is mounted a sonic flare tip with design capacity of 416,800 kg/h. Introduction Phase II HP flare system, comprising a duplex stainless steel Pipe collection system, carbon steel KO drum, carbon steel riser and stack on which is mounted a sonic flare tip with design capacity of 416,800 kg/h. Calculations of depressurization system reveal whether high volumes of gases will be flowing through the pipe header to the flare. In some cases, simultaneously depressurizing all the process and equipment, vessels, and piping in a plant will be difficult to accomplish (due to pipe sizing and economic impacts). In these cases, sequential or segmented depressurization of vessels should be considered. Providing for the worst vessels first or controlling the system to depressurize the area most affected first are possible scenarios that can be employed [11-14]. This paper discusses two options suggested when connecting a new facility to an existing process plant considering tie-in both flares systems into the existing. The available options for Flare tie-in are sequenced flare to PH-1 or PH-2 existing, flare combination to both PH-1 and/ or PH-2 and independent new compression flare utility. These options are competitive to find the feasible method for atmospheric disposal of the compression area (gas compressors and condensate suction drum) to HP flare blowdown loads to achieve the following objectives: - Case study Qasr is a large, normally pressured gas-condensate reservoir located in the Western Desert of Egypt approximately 525 km west of Cairo. The field is operated by Khalda Petroleum Company (KPC), a joint venture between Apache Corporation and Egyptian General Petroleum Company (EGPC). • Overpressure protection for process and utility systems • Means of automatic depressurizing of the Compression Project process facilities during emergency or upset conditions. Field production is initially handled at the Start of Field production is initially handled at the Start of 5796 PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 • Safe disposal of hydrocarbons which may - from time to time - be vented from the compression plant during abnormal operations and plant maintenance, start-up or shutdown. www.psychologyandeducation.net Compression areas. The plant layout is arranged into individual sections can be considered as separate fire zones. A credit will be taken for the area where the highest risk is occurred to be depressurized firstly then after the area under lower risk. This allows one section to be affected by a fire, without their necessarily being an impact on other sections. • Allow controlled manual depressurization of equipment items for maintenance, inspection and testing. • Minimize the potential pollution of the environment. Phase-1 process facility consists of 12 BDVs used in between the emergency shutdown valves dedicated to be used in emergency shutdown to release the process plant facility hazardous hydrocarbon. These tie-in options will compatible with one of the following blowdown scenarios: These tie-in options will compatible with one of the following blowdown scenarios: These tie-in options will compatible with one of the following blowdown scenarios: Conventional blowdown “Total plant blowdown starting from max. operating pressure reaching to 7 barg within 15 min”. Sequential blowdown starting from max. operating pressure reaching to 7 barg within 15 min., applying the sequential blowdown philosophy between Phase 1, Phase 2 and These tie-in options will compatible with one of the following blowdown scenarios: Conventional blowdown “Total plant blowdown starting from max. operating pressure reaching to 7 barg within 15 min”. Sequential blowdown starting from max. operating pressure reaching to 7 barg within 15 min., applying the sequential blowdown philosophy between Phase 1, Phase 2 and Conventional blowdown “Total plant blowdown starting from max. operating pressure reaching to 7 barg within 15 min”. Sequential blowdown starting from max. operating pressure reaching to 7 barg within 15 min., applying the sequential blowdown philosophy between Phase 1, Phase 2 and Phase-2 process facility consists of 7 BDVs used in between the emergency shutdown valves dedicated to be used in emergency shutdown to release the process plant facility hazardous hydrocarbon. The process facility emergency shutdown valves and blowdown valves is described in Figure 1 for Phase-1 and Figure 2 for Phase-2. 5797 www.psychologyandeducation.net www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 Compression facilities are new facilities added to the existing process facilities (Phase-1 and Phase-2 facilities) to increase the system pressure. These facilities are collected in a compression unit (Figure 4, a), a condensate handling system (Figure 4, b) and fuel system (Figure 4, c) . The condensate system consists of two identical suction drums A (cond. Methodology or phase-2 have reached to 7 barg at 15 minutes. Connection of compression facilities to phase-1 or phase-2 flare system depends on the design capacity of the flaring system (416,800 kg/h). Scenario-1: Implementation an independent new Compression Flare system The blowdown philosophy complies with the API 521 standards to release the hazardous hydrocarbon gas from the maximum pressure at 131 barg during the emergency to be reduced in 15 minutes to reach 7 barg. In such case the process facility can be maintained safe in case of emergency fire or even in the preliminary gas release. Compression areas. Drum A) and B (cond. Drum B) units, each in equal capacities and used in 100% redundancy as one in service and the second one is standby which is assumed to be depressurized and under positive nitrogen pressure to be preserved free of Compression facilities are new facilities added to the existing process facilities (Phase-1 and Phase-2 facilities) to increase the system pressure. These facilities are collected in a compression unit (Figure 4, a), a condensate handling system (Figure 4, b) and fuel system (Figure 4, c) . The condensate system consists of two identical suction drums A (cond. Drum A) and B (cond. Drum B) units, each in equal capacities and used in 100% redundancy as one in service and the second one is standby which is assumed to be depressurized and under positive nitrogen pressure to be preserved free of hydrocarbon liquid and gases. The fuel system contains a scrubber to free the gas from any entrained liquids associated with the gas. Also, there are preheaters to raise the gas stream temperature in a preparation step for conditioning the gas to be subjected to pressure reduction without hydrate formation. The new compression unit facility consists of 12 BDVs. These valves are used in between the emergency shutdown valves to release the process plant facility hazardous hydrocarbon. 5798 www.psychologyandeducation.net www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 Methodology Scenario-1: Implementation an independent new Compression Flare system Results and discussion The simulation is built using Aspen HYSYS program to perform the process plant facility blowdown scenarios in case of fire. The process plant facilities are simulated using Aspen Tech process modeling HYSYS V.10 and corresponding Aspen Flare System Analyzer V 10 for simulating the relieved gas different scenarios. The fire scenario is based on API-521 standard fire case. Each individual process equipment is to be sized according to the mechanical data sheet. Each pipe is to be specified according to the metal type, schedule and the nominal diameter and hence to state the pipeline length to determine the proposed volume of that pipe. In this suggested scenario, a new independent compression flare system will be constructed. The compression initial pressure assumed to be the maximum operating pressure for all equipment. As a result of pressure reduction at the compression suction header, the depressuring pressure will vary according to the operating condition. The blowdown philosophy applied to phase-1 and phase-2 is also applied to the new compression flare system. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing Scenario-1: Implementation an independent new Compression Flare system Scenario-1: Implementation an independent new Compression Flare system This scenario is based on arranging the plant flare systems into individual sections. These sections can be considered as separate fire zones. This allows one section to be affected by a fire, without an impact on the other sections. This design limits the requirement of full plant blowdown, which is desirable because an emergency blowdown results in the loss of a significant mass of hydrocarbons to the atmosphere (more conservative for HC gas). This sequenced scenario depends on relieving the compression facilities to phase-1 or phase-2 flare system after the piping and equipment of phase-1 The simulation is built using Aspen HYSYS v. 10 to perform the process plant facility blowdown scenario in case of fire through phase-1 and phase- 2 facilities. The fire scenario is based on API-521 standard fire case. Total flared gas during blowdown equals the sum of all BDVs Blowdown valves individually in each phase. All system depressuring in the moment of emergency case activation are taken into consideration to achieve the depressuring through 15 minutes. The total peak flow calculated for phase-1 is illustrated in Table 1 and for phase-2 is illustrated in Table-2: Table 1: Total flared gas during blowdown for phase 1 (scenario-1) 5799 www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 EBDV TAG SERVICE TOTAL VOLUME; Initia l Press . Initial Temp. Final Pressu re Orifice Area/Diam eter Blowdown Rate Gas m3 Liq. m3 Barg C Barg mm2/mm Kg/hr. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing 40-BDV-015 FROM FLOW LINE RECEIVER PACKAGE, PRODUCTION HEADER PACKAGE, TO AIR COOLERS 33.5851 131 90 7 776.4 / 31.44 46377 26.816 6.7691 40-BDV-016 FROM INLET TO AIR COOLERS, TEST SEPARATOR, TO INLET OF PRODUCTION SEPARATORS 61.38 131 90 7 1403 / 42.26 83798 49.02 12.36 40-BDV- 018A PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV- 018B PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV-004 From outlet of Production Separators to Phase -2 battery limit ESV at Salam Pipeline 14.613 131 57.8 7 371.3 / 21.74 24600 14.61 0.003 80-BDV-013 (new valve) INTERPIPING BETWEEN COMP. DISCHARGE & TIE IN 47.6795 131 57.8 7 1137 / 38 73139 47.67 0.0095 EBDV TAG SERVICE TOTAL VOLUME; Initia l Press . Initial Temp. Final Pressu re Orifice Area/Diam eter Blowdown Rate Gas m3 Liq. m3 Barg C Barg mm2/mm Kg/hr. 30-BDV- 037 Process gas outlet piping 25.995 0.005 80-BDV- 012 Inter-piping between Comp. Discharge & tie- in 36.304 131 57.8 7 873/ 33.34 56142 36.304 0 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr. 417150 Flare Designed Flow Rate, Kg/hr. 416800 Excess of Design % 0% Shortage of Design % -0.08% Table 2: Total flared gas during blowdown for phase 2 (scenario-1) EBDV TAG SERVICE TOTAL VOLUME; Initial Press. Initial Temp . Final Pressur e Orifice Area / Diamet er Blowdow n Rate Gas M3 Liq. M3 Barg oC Barg mm2/m m Kg/hr. 40-BDV-015 FROM FLOW LINE RECEIVER PACKAGE, PRODUCTION HEADER PACKAGE, TO AIR COOLERS 33.5851 131 90 7 776.4 / 31.44 46377 26.816 6.7691 40-BDV-016 FROM INLET TO AIR COOLERS, TEST SEPARATOR, TO INLET OF PRODUCTION SEPARATORS 61.38 131 90 7 1403 / 42.26 83798 49.02 12.36 40-BDV- 018A PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV- 018B PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV-004 From outlet of Production Separators to Phase -2 battery limit ESV at Salam Pipeline 14.613 131 57.8 7 371.3 / 21.74 24600 14.61 0.003 80-BDV-013 (new valve) INTERPIPING BETWEEN COMP. DISCHARGE & TIE-IN 47.6795 131 57.8 7 1137 / 38 73139 47.67 0.0095 EBDV TAG SERVICE TOTAL VOLUME; Initia l Press . Initial Temp. Final Pressu re Orifice Area/Diam eter Blowdown Rate Gas m3 Liq. m3 Barg C Barg mm2/mm Kg/hr. 30-BDV- 037 Process gas outlet piping 25.995 0.005 80-BDV- 012 Inter-piping between Comp. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing 30-BDV- 031 FROM FLOW LINES THROUGH PRODUCTION HEADER TO AIR COOLERS INLET 33.5851 131 110 7 762.2 / 31.15 43300 26.81 6 6.7691 30-BDV- 032 FROM FLOW LINES THROUGH TEST HEADER TO TEST SEPARATOR INLET 2.2784 131 90 7 60 / 8.73 3581 2.21 0.0684 30-BDV- 033 AIR COOLERS WITH INLET /OUTLET AND BYPASS LINE 11.7 131 92.3 7 279.3 / 18.86 16574 11 0.7 30-BDV- 034 AIR COOLERS WITH INLET /OUTLET AND BYPASS LINE 8.1 131 58 7 208.5 / 16.29 13750 7.6 0.5 30-BDV- 035 INTERCONNECTING PIPING BETWEEN AIR COOLERS & PRODUCTION SEPARATORS 17.63 131 57.8 7 441.7/ 23.71 29234 16.51 1.12 30-BDV- 602 Production Separator A 36.82 131 57.8 7 883.9/ 33.55 56846 29.456 7.364 30-BDV- 702 Production Separator B 36.82 131 57.8 7 883.9 / 33.55 56846 29.456 7.364 30-BDV- 802 Production Separator C 36.82 131 57.8 7 883.9 / 33.55 56846 29.456 7.364 30-BDV- 505 Test Separator 9.1 131 57.8 7 226.5 / 16.9 14553 7.8 1.3 30-BDV- 038 Process gas outlet piping 18.23 131 57.8 7 448.2 / 23.89 28825 18.23 0 26 131 57.8 7 632.1 / 28.37 40653 5800 5800 www.psychologyandeducation.net www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 EBDV TAG SERVICE TOTAL VOLUME; Initia l Press . Initial Temp. Final Pressu re Orifice Area/Diam eter Blowdown Rate Gas m3 Liq. m3 Barg C Barg mm2/mm Kg/hr. 30-BDV- 037 Process gas outlet piping 25.995 0.005 80-BDV- 012 Inter-piping between Comp. Discharge & tie- in 36.304 131 57.8 7 873/ 33.34 56142 36.304 0 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr. 417150 Flare Designed Flow Rate, Kg/hr. 416800 Excess of Design % 0% Shortage of Design % -0.08% Table 2: Total flared gas during blowdown for phase 2 (scenario-1) EBDV TAG SERVICE TOTAL VOLUME; Initial Press. Initial Temp . Final Pressur e Orifice Area / Diamet er Blowdow n Rate Gas M3 Liq. M3 Barg oC Barg mm2/m m Kg/hr. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing Discharge & tie- in 36.304 131 57.8 7 873/ 33.34 56142 36.304 0 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr. 417150 Flare Designed Flow Rate, Kg/hr. 416800 Excess of Design % 0% Shortage of Design % -0.08% Table 2: Total flared gas during blowdown for phase 2 (scenario-1) EBDV TAG SERVICE TOTAL VOLUME; Initial Press. Initial Temp . Final Pressur e Orifice Area / Diamet er Blowdow n Rate Gas M3 Liq. M3 Barg oC Barg mm2/m m Kg/hr. 40-BDV-015 FROM FLOW LINE RECEIVER PACKAGE, PRODUCTION HEADER PACKAGE, TO AIR COOLERS 33.5851 131 90 7 776.4 / 31.44 46377 26.816 6.7691 40-BDV-016 FROM INLET TO AIR COOLERS, TEST SEPARATOR, TO INLET OF PRODUCTION SEPARATORS 61.38 131 90 7 1403 / 42.26 83798 49.02 12.36 40-BDV- 018A PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV- 018B PRODUCTION SEPARATOR 30.69 131 57.8 7 742.1 / 30.74 47725 15.34 15.35 40-BDV-004 From outlet of Production Separators to Phase -2 battery limit ESV at Salam Pipeline 14.613 131 57.8 7 371.3 / 21.74 24600 14.61 0.003 80-BDV-013 (new valve) INTERPIPING BETWEEN COMP. DISCHARGE & TIE-IN 47.6795 131 57.8 7 1137 / 38 73139 47.67 0.0095 Table 2: Total flared gas during blowdown for phase 2 (scenario-1) 5801 www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 80-BDV-014 (new valve) INTERPIPING BETWEEN phase-2 -2 & TIE-IN 15.122 131 57.8 7 375.6/ 21.87 24154 15 0.122 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr. 34751 8 Flare Designed Flow Rate, Kg/hr. 416800 Excess of Design % 17% Shortage of Design % 0.00% suction header, the depressuring pressure will vary according to the operating condition. The total peak flow calculated as described in Table 3: The compression initial pressure assumed to be the maximum operating pressure for all equipment, as a result of pressure reduction at the compression 5 Table 3: Total flared gas during blowdown for the new compression unit EBDV TAG SERVICE TOTAL VOLUME ; Initial Press. Initial Temp. Final Pressure Orifice Area / Diam. Blowdo wn Rate Gas M3 Liq. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing M3 Barg oC Barg mm2/mm Kg/hr 80-BDV- 010 SEPARATORS OUTLET TO COMPRESSION SUCTION 123.865 85 47.58 7 2457/ 55.9 101223 123.86 5 0 80-BDV- 101 COMPRESSION MACHINE TRAIN A 62.3 95 72.76 7 1229/ 39.56 52350 57.8 4.4747 80-BDV- 201 COMPRESSION MACHINE TRAIN B 47.3951 100.23 76.71 7 956.4 / 34.89 42646 44.55 2.8451 80-BDV- 301 COMPRESSION MACHINE TRAIN C 62.2747 95 72.76 7 1229/ 39.56 52350 57.8 4.4747 80-BDV- 401 COMPRESSION MACHINE TRAIN D 47.3951 100.23 76.71 7 956.4 / 34.89 42646 44.55 2.8451 80-BDV- 011 COMPRESSION DISCHARGE TO EXISTING EXPORT FACITILIES 58 130 57.8 7 1370 / 41.76 87377 58 0 80-BDV- 001A CONDENSATE SUCTION DRUM A 28.165 70 57.8 7 511.8 / 25.53 8196.5 19.595 8.57 80-BDV- 001B CONDENSATE SUCTION DRUM B 28.194 70 57.8 7 511.8 / 25.53 8196.5 19.624 8.57 Table 3: Total flared gas during blowdown for the new compression unit 5802 PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 EBDV TAG SERVICE TOTAL VOLUME ; Initial Press. Initial Temp. Final Pressure Orifice Area / Diam. Blowdo wn Rate Gas M3 Liq. M3 Barg oC Barg mm2/mm Kg/hr 80-BDV- 002 FLASH GAS PREHEATER 0.068 70 57.8 7 1.264 / 1.269 40.49 0.068 0 80-BDV- 004A HP FUEL GAS PREHEATER 2.304 130 57.44 7 59.15 / 8.67 1889 2.304 0 80-BDV- 004B HP FUEL GAS PREHEATER 2.258 130 57.44 7 59.15 / 8.67 1889 2.258 0 80-BDV- 006 HP FUEL GAS KO DRUM 7.291 42 50 7 103.3 / 11.47 1935 7.291 0 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr 400738.4 9 Flare Designed Flow Rate, Kg/hr 416800 Excess of Design % 4% Shortage of Design % 0.00% capacity by 17% of design capacity 416800 Kg/hr.). The total flared gas directed to an identical new flare will be 400738 Kg/hr. (which is lesser than the design flare capacity by 4%). The new identical flare system estimated cost with +/- 10% accuracy is illustrated in Table 4. The results above show that the total flared gas directed to phase-1 flare will be 417150 Kg/hr. (it exceeds the design flare capacity by 0.08% of design capacity 416800 Kg/hr.) which is accepted considering the flare design margin. The total flared gas directed to phase-2 flare will be 347518 Kg/hr. the phase-1 flare design requirements. the phase-1 flare design requirements. 2- Once the compression facility starts to relief to phase-2 flare, after Phase-2 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by phase- 2 flare for the following 15 min. is 415604.49 Kg/hr (Table 6). (which is lesser than the design flare capacity by 0.3% of flare design capacity 416800 Kg/hr.). That means that the sequential blowdown solution will meet the phase-2 flare design requirements and also the blowdown criteria. 2- Once the compression facility starts to relief to phase-2 flare, after Phase-2 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by phase- 2 flare for the following 15 min. is 415604.49 Kg/hr (Table 6). (which is lesser than the design flare capacity by 0.3% of flare design capacity 416800 Kg/hr.). That means that the sequential blowdown solution will meet the phase-2 flare design requirements and also the blowdown criteria. Simultaneously depressurizing all the process and equipment, vessels, and piping in a plant will be difficult to accomplish (due to pipe sizing and economic impacts). In these cases, sequential or segmented depressurization of vessels should be considered. Providing for the worst vessels first or controlling the system to depressurize the area most affected first are possible scenarios that can be employed. Sequential blowdown starts from equipment maximum operating pressure to 7 Barg within 15 minutes. This scenario is performed in order to estimate the peak blowdown rate for each zone based on external fire depressuring criteria and to depressurize to 7 Barg in 15 minutes: 3- Adding the new compression facility considering the sequenced flaring technique will cost a new system of uninterrupted power supply (UPS) with logic solver to resume efficient in case of total plant power failure. Use of UPS or emergency diesel generator may be considered so that when the normal power supply is interrupted, a standby power supply would automatically start, in a fraction of a second, to support critical equipment or units. Estimated cost with +/- 10% accuracy for new redundant UPS system to avoid any failure causing direct flaring at the same time is approximately 10000 $. Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing (which is lesser than the design flare Table 4: New Identical Flare System Cost Estimation Equipment Cost ($) 200 m Piping (16” Duplex): Headers, sub headers and supports 400,000 Carbon Steel HP Flare Knock Out Drum 1,000,000 Carbon Steel HP Flare Stack 1,000,000 Total 2,400,000 Table 4: New Identical Flare System Cost Estimation not be affected when one of the existing flares is under maintenance. But on the other hand, will negatively affect the project budget as this requires a high capital cost of the new flare system. Adding a new independent compression flare utility will positively help relieving the new compression surge volume in a minimum time as per API521 standard (15 minutes or less). Independent flare will not affect existing facilities depressuring time and in addition the project will 5803 www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing Scenario-2: Sequenced Flare to Phase-1 and Phase-2 existing the phase-1 flare design requirements. The results of scenario 2 show that: 1- Once the compression facility starts to relief to phase-1 flare after phase-1 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by phase- 1 flare for the following 15 minutes is 418812.49 Kg/hr (Table 5). (which exceeds with accepted limits the design flare capacity of PH-1 by 0.5% of flare design capacity 416800 Kg/hr.). That means that the sequential blowdown solution will not meet 1- Once the compression facility starts to relief to phase-1 flare after phase-1 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by phase- 1 flare for the following 15 minutes is 418812.49 Kg/hr (Table 5). (which exceeds with accepted limits the design flare capacity of PH-1 by 0.5% of flare design capacity 416800 Kg/hr.). That means that the sequential blowdown solution will not meet 58 h l d d Table 5: Total Flared Gas during Blowdown for phase-1 (scenario- 2) EBDV TAG SERVICE TOTAL VOLUME; Initial Press. Initial Temp . Fin al Pre ss. Orifice Area/Diam eter Blowdo wn Rate Balanc e Flow After 15 min GAS M3 LIQ. M3 Barg C Bar g mm2/mm Kg/hr 30-BDV- 031 FROM FLOW LINES THROUGH PRODUCTION HEADER TO AIR COOLERS INLET 33.5851 131 110 7 762.2 / 31.15 43300 1895 26.81 6 6.769 1 30-BDV- 032 FROM FLOW LINES THROUGH TEST HEADER TO TEST SEPARATOR INLET 2.2784 131 90 7 60 / 8.73 3581 144.6 2.21 0.068 4 11.7 131 92.3 7 16574 662.4 Table 5: Total Flared Gas during Blowdown for phase-1 (scenario- 2) PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 EBDV TAG SERVICE TOTAL VOLUME; Initial Press. Initial Temp . Fin al Pre ss. Orifice Area/Diam eter Blowdo wn Rate Balanc e Flow After 15 min GAS M3 LIQ. the phase-1 flare design requirements. M3 Barg C Bar g mm2/mm Kg/hr 30-BDV- 033 AIR COOLERS WITH INLET /OUTLET AND BYPASS LINE 11 0.7 279.3 / 18.86 30-BDV- 034 AIR COOLERS WITH INLET /OUTLET AND BYPASS LINE 8.1 131 58 7 208.5 / 16.29 13750 531.8 7.6 0.5 30-BDV- 035 INTERCONNECTING PIPING BETWEEN AIR COOLERS & PRODUCTION SEPARATORS 17.63 131 57.8 7 441.7 / 23.71 29234 1145 16.51 1.12 30-BDV- 602 Production Separator A 36.82 131 57.8 7 883.9 / 33.55 56846 2512 29.45 6 7.364 30-BDV- 702 Production Separator B 36.82 131 57.8 7 883.9 / 33.55 56846 2512 29.45 6 7.364 30-BDV- 802 Production Separator C 36.82 131 57.8 7 883.9 / 33.55 56846 2512 29.45 6 7.364 30-BDV- 505 Test Separator 9.1 131 57.8 7 226.5 / 16.9 14553 680.2 7.8 1.3 30-BDV- 038 Process gas outlet piping 18.23 131 57.8 7 448.2 / 23.89 28825 1245 18.23 0 30-BDV- 037 Process gas outlet piping 26 131 57.8 7 632.1 / 28.37 40653 1769 25.99 5 0.005 80-BDV- 012 Inter-piping between Comp. Discharge & tie- in 36.304 131 57.8 7 873 / 33.34 56142 2465 36.30 4 0 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr 417150 18074 Flare Designed Flow Rate, Kg/hr 416800 416800 Excess of Design % 0% Shortage of Design % -0.08% Required Compression Flared Gas 400738 .49 In Case of Sequential Blowdown, Total Compression Flared Gas 418812 .49 In Case of Sequential Blowdown, Excess of Design % 0.0% In Case of Sequential Blowdown, Shortage of Design % -0.5% PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 5806 h l d d i Table 6: Total Flared Gas during Blowdown for Phase-2 (scenario-2) EBDV TAG SERVICE TOTAL VOLUME; Ini tial Pr ess . Init ial Te mp . Fi nal Pr ess . Orifice Area/ Diamet er Blow down Rate Bala nce Flow After 15 min Gas m3 Liq. m3 Ba rg C Ba rg mm2/m m Kg/hr . Kg/h r. 40- BDV- 015 FROM FLOW LINE RECEIVER PACKAGE, PRODUCTION HEADER PACKAGE, TO AIR COOLERS 33.56 13 1 90 7 776.4/ 31.44 4637 7 1967 26.8 6.76 40- BDV- 016 FROM INLET TO AIR COOLERS, TEST SEPARATOR, TO INLET OF PRODUCTION SEPARATORS 61.38 13 1 90 7 1403/ 42.26 8379 8 3523 49.0 2 12.3 6 40- BDV- 018A PRODUCTION SEPARATOR 30.6 13 1 57. the phase-1 flare design requirements. 8 7 742.1 / 30.74 4772 5 2086 15.3 15.3 40- BDV- 018B PRODUCTION SEPARATOR 30.6 13 1 57. 8 7 742.1 / 30.74 4772 5 2086 15.3 15.3 40- BDV- 004 From outlet of Production Separators to Phase-2 battery limit ESV at Salam Pipeline 14.603 13 1 57. 8 7 371.3 / 21.74 2460 0 968 14.6 0.00 3 80- BDV- 013 (NEW VALV E) INTERPIPING BETWEEN COMP. DISCHARGE & TIE- IN 47.609 13 1 57. 8 7 1137 / 38 7313 9 3221 47.6 0.00 9 80- BDV- 014 (NEW VALV E) INTERPIPING BETWEEN PH II & TIE-IN 15.12 13 1 57. 8 7 375.6 / 21.87 2415 4 1015 15 0.12 TOTAL FLARED GAS DURING BLOWDOWN, Kg/hr 3475 18 1486 6 Flare Designed Flow Rate, Kg/hr 4168 00 4168 00 Excess of Design % 17% Shortage of Design % 0.00 % Required Compression Flared Gas 4007 38.49 Table 6: Total Flared Gas during Blowdown for Phase-2 (scenario-2) 5806 38.49 www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 EBDV TAG SERVICE TOTAL VOLUME; Ini tial Pr ess . Init ial Te mp . Fi nal Pr ess . Orifice Area/ Diamet er Blow down Rate Bala nce Flow After 15 min Gas m3 Liq. m3 Ba rg C Ba rg mm2/m m Kg/hr . Kg/h r. Total Compression Flared Gas 4156 04.49 In Case of Sequential Blowdown, Excess of Design % 0.3% In Case of Sequential Blowdown, Shortage of Design % 0.0% flare design margin). Once the compression facility starts to relief to Phase-2 flare, after Phase-2 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by Phase-2 flare for the following 15 min. is 415604.49 Kg/hr. (which is lesser than the design flare capacity by 0.3% of flare design capacity 416800 Kg/hr. Adding the new compression facility considering the sequenced flaring technique will cost a new system of UPS with logic solver to resume efficient in case of total plant power failure, it approximately costs 10000 $. Conclusion The two scenarios for connecting a new flare system of the new compression unit to the two existing flare systems are considerd. The two scenarios are implementing a new flare system for the new compression unit and sequential blowdown,. Simulation and calculations revealed that: For scenario 1, if a new flare system is used (Final Pressure 7 barg within 15 min) the total flared gas will be directed to Phase-1 flare will be 417150 Kg/hr. (it exceeds the design flare capacity by 0.08% of design capacity 416800 Kg/hr.) which is accepted considering the flare design margin, and the total flared gas will be directed to Phase-2 flare will be 347518 Kg/hr. (which is lesser than the design flare capacity by 17% of design capacity 416800 Kg/hr.). For compression depressurization, the total flared gas will be directed to an identical new flare will be 400738 Kg/hr (which is lesser than the design flare capacity by 4%). A new flare for the peak load of 400738 Kg/hr will require a large capital cost which is approximately 2,400,000 $. References [1] Karl Kolmetz, Riska Ristiyanti, Kolmetz Handbook of Process Equipment Design FLARE SYSTEMS SAFETY, SELECTION AND SIZING AND TROUBLESHOOTING (ENGINEERING DESIGN GUIDELINE), KLM Technology Group Practical Engineering Guidelines for Processing Plant Solutions, 2015 [2] Carbon Limits AS., Flare Reduction in Egypt report, 15.04.2016. For scenario 2, if sequential blowdown (final Pressure 7 Barg in 15 min.) is used, Once the compression facility starts to relief to Phase-1 flare after Phase-1 piping and equipment have reached to 7 barg at 15 min, the total flared gas will be handled by Phase-1 flare for the following 15 min. is 418812.49 Kg/hr. (which exceeds the design flare capacity by 0.5% of flare design capacity 416800 Kg/hr. which is accepted considering the [3] Mikhail Zhizhin, Kimberly Baugh, Feng- Chi Hsu CIRES, Results from a Survey of Global Natural Gas Flaring from Visible Infrared Imaging Radiometer Suite Data, Chris Elvidge, Earth observation group, NOAA National Centers for Environmental Information, May 18, 2016. [4] 5807 [4] ADAM BADER, CHARLES E.BAUKAL, JR., P.E., WES BUSSMAN, Selecting the www.psychologyandeducation.net PSYCHOLOGY AND EDUCATION (2021) 58(1): 5795-5808 ISSN: 00333077 proper flare system, American Institute of Chemical Engineers, 2011 [5] Daniel A. Crowl, Joseph F. Louvar, Chemical Process Safety, Fundamentals with Applications, 3rd edition ,2011 [6] Douglas Attawy, David A. Coyle, GROUND FLARES – OUT OF SIGHT, OUT OF MIND. LNG Industry, PERTH AUSTRALIA, 11-15 April 2016. [7] Ken Arnold, Maurice Stewart, Surface Production Operations, Volume 2: 2nd Edition, Design of Gas-Handling Systems and Facilities, Gulf Publishing Company, 1999. [8] Nicholas P. Cheremisinoff, Industrial Gas Flaring Practices, PhD, John Wiley & Sons Inc, 2013. [9] Pressure-relieving and depressuring systems, ANSI/API STANDARD 521, FIFTH EDITION, JANUARY 2007 (INCLUDES ERRATA JUNE 2007). [10] Frank Lees, Lee’s Loss Prevention in the Process Industries, 3rd Edition, Volume 1, Butterworth-Heinemann, 2005. [11] Flare Details for General Refinery and Petrochemical Service, ANSI/API STANDARD 537, SECOND EDITION, DECEMBER 2008. [12] Don W. Green, Robert H. Perry, Perry's Chemical Engineers' Handbook, 8th Edition, McGraw-Hill Companies, Inc., 2008 [13] Dennis P. Nolan, Handbook of Fire and Explosion Protection Engineering Principles: for Oil, Gas, Chemical and Related Facilities, 4th edition, Gulf Professional Publishing 2019. [14] Charles E. Baukal Jr., The John Zink Hamworthy Combustion Handbook : Fundamentals (Industrial Combustion), Volume 1 , 2nd edition, CRC Press, 2012 5808 www.psychologyandeducation.net	32,963
62/hal.inria.fr-hal-00913639-document.txt_1	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	367	590	Musique T ouCheR la CoRde seNsible Contrairement aux apparences, le piano est un système acoustique et mécanique sophistiqué dont la facture demeure à ce jour très largement empirique. Les connaissances, très précises, des concepteurs et fabricants de pianos sont issues de siècles d'expérimentations, d'échecs, de succès Et intriguent beaucoup les chercheurs en acoustique musicale. En utilisant des méthodes scientifiques pour modéliser le fonctionnement d'un piano et de ses différents éléments, il est possible de confirmer ou non ces connaissances et d'aller plus loin dans la compréhension des phénomènes mis en jeu. Chacune des étapes acoustiques intervenant lors de l'exécution d'une note (voir encadré) peut être modélisée par une équation particulière. C'est le travail qui a été mené par Juliette Chabassier, d'Inria : elle a établi et discrétisé (découpé en tout petits intervalles de temps et d'espace) un modèle général puis calculé numériquement les solutions grâce à des méthodes spécialement développées. « Ce travail s'inscrit dans le cadre de mon doctorat en mathématiques appliquées co-encadré par Patrick Joly, directeur du projet INRIA POEMS et spécialiste de modélisation et analyse numérique pour la propagation des ondes, et Antoine Chaigne, directeur de l'Unité de Mécanique de l'ENSTA et spécialiste d'acoustique musicale », précise-t-elle. Grâce à ce modèle et aux simulations numériques qui en découlent, la jeune scientifique est parvenue à reprodu fidèlement des formes expérimentales, mais aussi, par exemple, à connaître le champ de pression de l'air sur un ensemble de points tout De façon schématique, lorsqu'une touche du clavier est jouée, un mécanisme très précis dé- autour du piano, impossible à mesurer dans la réalité sans perturber le système. multiplie son mouvement et actionne un mar« L'une des applications de ce modèle et de sa discrétisation numéteau qui frappe entre une et trois cordes à la rique est bien sûr l'aide à la facture instrumentale pour valider le savoir fois (selon la note choisie). Les cordes se metmillénaire et empirique tent alors en vibration et c'est le chevalet qui des concepteurs et fabritransmet l'énergie des cordes à la table d'harmonie, qui vibre elle aussi, mettant en mouve- cants de pianos mais aussi approfondir notre comment les molécules d'air avoisinantes, et préhension des phénoentrainant la propagation d'un son dans l'air.	3,335
https://openalex.org/W4223979515	OpenAlex	Open Science	CC-By	2,022	Political Thought and the Struggle for Sovereignty in Ethiopian-Japanese Relations (1927–1936)	Sara Marzagora	English	Spoken	13,015	19,249	Link to publication record in King's Research Portal Citation for published version (APA): Marzagora, S. (2023). Political Thought and the Struggle for Sovereignty in Ethiopian-Japanese Relations (1927–1936). INTERNATIONAL HISTORY REVIEW, 45(1), 95-113. https://doi.org/10.1080/07075332.2022.2058586 Citing this paper Pl h C t g t s pape Please note that where the full-text provided on King's Research Portal is the Author Accepted Manuscript or Post-Print version this may differ from the final Published version. If citing, it is advised that you check and use the publisher's definitive version for pagination, volume/issue, and date of publication details. 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Download date: 25. Oct. 2024 The International History Review Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=rinh20 CONTACT Sara Marzagora sara.marzagora@kcl.ac.uk This article has been republished with minor changes. These changes do not impact the academic content of the article. 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. KEYWORDS Ethiopia; Japan; Ethiopian political thought; young Ethiopians; League of Nations; Abyssinian crisis; Pan-Asianism; sovereignty; economic dependency ABSTRACT Ethiopia and Japan tightened their diplomatic and economic relationships between 1927 and 1931. Haile Selassie and the “progressive” intellectuals around him were striving to strengthen Ethiopia’s claim to sovereignty. Japan as an example of non-white success added ammunition to their claim that Ethiopia had a right to sit at the table of the "great nations". The Japanese government sought to expand its trade, and probably understood its economic penetration in Ethiopia as a tentative imperial foothold in the African continent. The two governments never envisioned the alliance as a substitute for their respective pro-Western foreign policy, but sectors of their societies thought otherwise. Both the Young Ethiopians and the Japanese Pan-Asianists were critical of their respective governments’ capitalist internationalism and thought that the racism that shaped Western attitudes towards non-Europeans should be antagonised, not appeased. The analysis of the international context with which Ethiopian elites had to contend allows us to recontextualise and reframe the “alienation debate” in Ethiopian intellectual history. The “turn to Japan” should not be understood as an xenophilic infatuation with a fash- ionable foreign model, but on the contrary as a sophisticated and calcu- lated response to the international relations of the time. Political Thought and the Struggle for Sovereignty in Ethiopian-Japanese Relations (1927–1936) Sara Marzagora To cite this article: Sara Marzagora (2022): Political Thought and the Struggle for Sovereignty in Ethiopian-Japanese Relations (1927–1936), The International History Review, DOI: 10.1080/07075332.2022.2058586 To link to this article: https://doi.org/10.1080/07075332.2022.2058586 © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group Published online: 14 Apr 2022. Submit your article to this journal Article views: 58 View related articles View Crossmark data © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group Published online: 14 Apr 2022. Submit your article to this journal Article views: 58 View related articles View Crossmark data THE INTERNATIONAL HISTORY REVIEW https://doi.org/10.1080/07075332.2022.2058586 Political Thought and the Struggle for Sovereignty in Ethiopian-Japanese Relations (1927–1936) Sara Marzagora Comparative Literature, King’s College London, London, UK Sara Marzagora Comparative Literature, King’s College London, London, UK Comparative Literature, King’s College London, London, UK 1. Ethiopia, sovereignty, and international law Scholarship on Ethiopian intellectual history has been dominated by the so-called “alienation debate”. The debate departed from the premise that there has been an intellectual failure in Ethiopian history from the beginning of the twentieth century onwards. The reason for this fail- ure, scholars have argued, lies in the derivative nature of Ethiopian political thought. Rather than charting a locally-rooted politico-economic model of development, Ethiopian intellectuals have always opted for imitating others. Christopher Clapham calls this “the politics of emulation”: “a series of attempts by ‘modernising’ Ethiopians to identify the mechanisms of developmental suc- cess of countries perceived as having some similarity to their own”.1 This imitative understanding of development has been considered evidence of “cultural dislocation”, in the words of Messay Kebede, or even more strongly “native colonialism”, in the words of Yirga Gelaw Woldeyes.2 y y The widespread admiration for Japan among the Ethiopian intellectuals of the beginning of the twentieth century has been interpreted as an early instance of such derivative conception of 2 S. MARZAGORA development. For Clapham, Japan was the “preferred paradigm” of the two or three decades before 1936. This was a period, as we shall see, in which Amharic print culture frequently fea- tured calls for Ethiopia to follow the Japanese model, to the point that the intellectuals active in the interwar period have become known in the scholarship as “the Japanisers”. Clapham argues that, like the other imitative trends that preceded it and followed it, the Japanese paradigm was “fatally flawed”.3 Its failure came from an inadequate knowledge of Japan and an overestimation of the similarities between Japan and Ethiopia. The model was aspirational but completely unviable in practice. Bahru Zewde agrees that “the impassioned pleas of the ‘Japanizers’ remained a subjective urge unsupported by objective reality”.4 Ethiopia simply did not have the “capacity to emulate either the technological or the organisational achievements of late nine- teenth century Japan”.5 In this article, I situate the thought of the Ethiopian “Japanisers” within the global politico- economic power relations of the interwar years. An analysis of the international context with which Ethiopian elites had to contend will allow me to recontextualise and reframe the “alienation debate” in Ethiopian intellectual history. The “turn to Japan” should not be under- stood as an idiosyncratic choice for an attractive foreign model, but on the contrary as a calcu- lated response to the international relations of the time. 1. Ethiopia, sovereignty, and international law Ethiopia entered the twentieth century as an independent polity, having defeated an Italian invasion attempt in 1896 at the Battle of Adwa. In the interwar years, it was, together with Liberia, the only formally independent polity in the African continent. Ethiopian elites were aware that this independence was fragile. Ethiopia was surrounded by Italian, French and British colonies, and each of the three colonisers had expansionist interests in Ethiopia, whether in terms of economic penetration or straight-up territorial annexation. The 1906 Tripartite Treaty signed by Italy, France and Britain divided Ethiopia into European spheres of influence, making it painfully clear to Ethiopian elites that the Europeans remained a threat.6 The Ethio-European bor- der negotiations of the first years of the twentieth century were signed in conjunction with trea- ties on extraterritoriality, which further inscribed Ethiopia’s subordinate position internationally. The 1908 Franco–Ethiopian Treaty of Amity and Commerce (also known as the “Klobukowski Treaty”) represented the “first meaningful attempt by Europeans to extend their extraterritorial jurisdiction into Ethiopia”.7 Although Ethiopia’s extraterritoriality regime was weaker than other extraterritoriality regimes, for example in East Asia, this should not be interpreted as a sign of Ethiopia’s international strength. On the contrary, according to Hailegabriel Feyissa, extraterritori- ality was weaker in Ethiopia precisely because European powers considered it a matter of time before Ethiopia fell into European hands. Capitalist integration also proceeded along deeply unequal lines. The Djibouti-Addis Ababa railway, initially a private concession to a French com- pany and later taken over by the French state without Emperor Menelik’s consent, is regarded as “one of the very few successful illustrations of European economic imperialism in early 20th-cen- tury Ethiopia”.8 Menelik tried to offset the greater influence France had gained through the rail- way by granting the British a concession to set up Ethiopia’s first bank. Charles Schaefer, who studied the history of the bank, concluded that in its first years the bank acted “as a tool of British imperialism”.9 Everybody who had personal, political or ideological stakes in Ethiopia’s government was apprehensive about Ethiopia’s political fragility on the international scene. Ethiopia should develop, intellectuals and public figures kept repeating, or otherwise risked losing its independ- ence. The development Ethiopia should pursue was framed in terms of “catching up with Europe”. 1. Ethiopia, sovereignty, and international law In fact, although the intellectuals of this period are known as “the Japanisers”, calls for Ethiopia to follow in Europe’s footsteps were infinitely more frequent in Amharic print culture than articles or books mentioning Japan. This already puts into discussion Clapham’s assessment that Japan was the “preferred paradigm” for Ethiopian intellectuals at the time. And “becoming more like Europe” was not just the xenophilic call of culturally alienated intellectuals. On the contrary, it was about boosting those political and economic credentials that could allow the THE INTERNATIONAL HISTORY REVIEW 3 3 Ethiopian elites to demonstrate they were perfectly able to make the country progress without European intervention. Most of the diplomatic squabbles between the Ethiopian government and European governments in these years revolved around the control of borders. European colonial administrators repeatedly complained to Emperor Menelik that pastoralist groups were crossing the borders from Ethiopia into European colonies for cattle raids or armed skirmishes, creating economic and political disturbances.10 The Ethiopian government was aware of what was at stake in those complaints: the inability to enforce border legislation could be constructed as evidence of the Ethiopian government’s failure to meet the standards of sovereign statehood. This, in turn, could be used by European powers as an excuse for an invasion. For Ethiopia to maintain its independence, a process of reforms had to start, and framed in the right language. In a Eurocentric international system that served white supremacist values, the criteria that determined sovereign statehood all derived from few Western European “prototypes”. Ethiopian political culture had traditionally operated according to different criteria. The old Ethiopian empire did not have a neatly demarcated territorial identity. Much like other pre-colonial African polities, the control over people and resources was much more important than the control over a set territory. While sovereignty was increasingly being defined in international law as the stable ability of an “effective” and “responsible” government to administer a “permanent population” and “a defined territory”,11 older power configurations in the Horn of Africa relied on a much more varied and layered typology of relations. 1. Ethiopia, sovereignty, and international law The Ethiopian Empire, for example, exacted trib- ute from other polities, but being “tributary to” did not necessarily mean “under the control of”.12 Terms like “subject to” and “tributary to”, argues Reid, are “frustrating and unsatisfying umbrella terms used to describe –and disguise– a wide range of political, diplomatic and military relationships, varying degrees of influence and/or control, and levels of short- and long-range regional suzerainty or hegemony”.13 From the modern period onwards, though, the global space was gradually reconfigured. International law worked to create a specific kind of individual and collective subjectivity that emerged out of, and in turn served to reproduce, the capitalist order. The non-European world was gradually “forced to reconstitute its myriad alternative normative orders and practices of self-organisation in this way or face the consequences – consequences which have ranged from outright colonisation to more recent forms of armed intervention”.14 This reconstitution worked via both coercion and desire: the new legal subjectivity was imposed at gunpoint or under the threat of military violence, but also framed as a “modernity” and “progress” that non-Europeans should aspire to.15 It is often difficult to separate the two: when Ethiopian intellectuals talked about the need to become “more like Europe” they aspired to a better society as much as they feared foreign conquest. Modernisation did not have to entail Westernisation, but in a country like Ethiopia, whose sovereignty was recognised only intermittently, modernisation could not mean something radically different from Westernisation either. Ethiopia could try to beat Europe at its own rules, but it was not in a political, economic or military position to set the rules. The encounter between Ethiopian emperors and international law introduced new conceptual frameworks for the exercise and legitimation of power, and Ethiopian emperors learnt to appro- priate the language of international law to boost their own political position. Menelik was aware that his international standing depended on making selected concessions to the new normative framework of sovereign statehood. Scholars that have traced the history of his diplomatic deal- ing with Europeans have traced precisely how, over decades, he gradually came to master the new international legal language to his advantage. After some initial diplomatic blunders, the Emperor realised that it was advantageous, if not straight-up fundamental for Ethiopia’s inde- pendence, to continue gesturing towards the international criteria for statehood. 1. Ethiopia, sovereignty, and international law Around the time of the Battle of Adwa, for example, Emperor Menelik engaged in sustained military cam- paigns to the south, south-west and south-east of the traditional highland core of the Ethiopian empire, using the presence of his men on the ground as evidence of “effective occupation” dur- ing his border negotiations with the three European powers that had annexed neighbouring 4 S. MARZAGORA territories.16 As Rose Parfitt has noted, these treaties did not imply the recognition, on the part of Europeans, of a pre-existing Ethiopian sovereignty, but rather it was the treaties themselves that constituted Ethiopia as a sovereign state and called into being a relationship of for- mal equality.17 territories.16 As Rose Parfitt has noted, these treaties did not imply the recognition, on the part of Europeans, of a pre-existing Ethiopian sovereignty, but rather it was the treaties themselves that constituted Ethiopia as a sovereign state and called into being a relationship of for- mal equality.17 Menelik was probably aware that his policy of concessions, especially when it came to key infrastructure (the railway) and economic institutions (the bank) essentially entailed outsourcing his sovereign prerogatives, but as long as he could play the three European powers off each other or off a fourth party, he could hope to minimise the dangers of dependency. Plus, there was more to be gained by the presence of a central bank than there was to be lost by the fact that this central bank was foreign-run. Railways, telegraph lines, banks were tools to integrate Ethiopia into the global capitalist economy, an integration that states recognised as sovereign were expected to embrace. In this sense, Menelik sought to defend Ethiopia’s independence by conceding to an international order which, despite the promise of “sovereign equality”, actually locked Ethiopia in a dependent position. At the same time, though, the Emperor was clear that subordination did not mean capitula- tion. Some of the concessions he made to European powers were formal and not substantive. In 1907, he formed Ethiopia’s first Council of Ministers, announcing to foreign powers that “it is some time since we thought of introducing a European system into our country. You have always [ … ] said it would be good if we [ … ] would adopt some of the European system. 1. Ethiopia, sovereignty, and international law I have now started to appoint a ministry”.18 De facto, though, the Council of Ministers operated like a traditional advisory body. The appointees “had only the vaguest ideas about their responsibilities” and Menelik himself “made only perfunctory use of the new ministry”.19 Already at the time of Menelik, then, the political debate in Addis Ababa was not “whether or not to ‘become more European’, but rather how to obtain those aspects of European existence which might allow the Empire to remain what it already was”.20 The challenge faced by Ethiopian Emperors was that international equality was predicated on sameness – on firmly renouncing Ethiopia’s political culture and transforming Ethiopia and its people according to the international definition of statehood and citizenry. Defending a degree of “otherness” was extremely dangerous, as non-compliance with the international legal order would be seen as a failure to meet the standards of sovereignty and civilisation. But as risky as it was, the Emperors had to defend a degree of “otherness”, because their domestic legitimacy in the eyes of the Ethiopian establishment depended on another epistemological framework – the political theology of the divinely-ordained monarchy. There was indeed a strong political faction in Ethiopia of church leaders and aristocrats (usually defined as “conservatives” in the scholar- ship) which strongly resisted the Emperor’s concessions to the international order and did not want to compromise on Ethiopia’s political identity. That Ethiopian Emperors could not rule with- out the support of this faction would become evident in 1916, when Menelik’s appointed heir Iyasu was deposed in a palace coup organised by the aristocracy, and in the end also validated by the head of the Ethiopian Orthodox Church. The coup makers disapproved of Iyasu’s redistri- bution of power away from Addis Ababa towards the eastern regions of the Empire and were also apprehensive about his pro-Ottoman leanings during the First World War.21 Ras Tafari, who was appointed heir to the throne under Empress Zawditu after the coup, had to contend with this political faction and their intransigent defence of Ethiopia’s traditional political culture, while also appeasing the international community’s demands for change and reform. The establishment of the League of Nations at the end of the First World War was celebrated by many within the Ethiopian government as a possible way out of the conundrum of politico- economic subordination. 1. Ethiopia, sovereignty, and international law Tafari saw the League as an institution that could create an inter- national level playing field, protecting small nations and restructuring international relations on more equal premises. He applied for membership on behalf of Ethiopia in September 1923. The League’s response showed that Tafari’s hopes were partly misplaced, as Ethiopia’s application was accepted only conditionally. Ethiopia had a right to remain a member of the League for as THE INTERNATIONAL HISTORY REVIEW 5 long as it worked towards meeting three “special obligations” related to the eradication of slav- ery and the slave trade. Ethiopia’s sovereignty was at the same time recognised and considered insufficient. The Ethiopian Empire “clearly bore some resemblances to the international legal con- ception of statehood. Yet it also continued to manifest certain other traits which European offi- cials and lawyers found impossible to ‘recognise’ as belonging to a state. [ … ] Ethiopia [ … ] was narrated, and constituted, quite literally, as a simultaneously sovereign and less-than-sover- eign entity”.22 Ethiopian intellectuals attempted to compensate for Ethiopia’s military, diplomatic, and eco- nomic weakness with a discursive offensive that cast their country as kith and kin of Europe on the grounds of its Christian past. Christianity had always been a central source of identity for the Ethiopian monarchy in previous centuries, and it could now be used as a useful weapon in the Ethiopian diplomatic arsenal. The League of Nation’s founding charter had been called a “covenant” precisely to evoke the Biblical notion of God’s covenant. Religion was a key source of identity for Western powers and a key mechanism through which Eastern “otherness” (Muslim, Hindu, Buddhist) was constructed. Ethiopia, though, could use religion as a powerful source of “sameness”. By stressing the Christian character of Ethiopian civilisation, the Ethiopian intelligent- sia claimed that Ethiopia naturally belonged to the club of the world’s “great nations”, despite not conforming to all international criteria for sovereign statehood. The bid for international acceptance was already perfectly summarised by Af€aw€arḳG€abr€a- Iyy€asus (1868-1947) in his 1908 Guide du Voyageur en Abyssinie: if Ethiopia was lagging behind Europe, one should “blame the governments in power up until this day in Ethiopia, a country so renowned and distinguished for its old traditions as a big Christian empire”.23 Ethiopia’s present underdevelopment was due to a series of accidental historical conjunctures, Af€aw€arḳargues, and did not reflect in any ways Ethiopia’s intrinsic civilisational standing. 1. Ethiopia, sovereignty, and international law It was the unfortunate suc- cession of bad leaders that held Ethiopia back, but in terms of its civilisational essence, Ethiopia was a renowned and distinguished Christian country, much like its European counterparts. Now that finally Ethiopia had a good ruler in Emperor Menelik, it was rapidly catching up with her bigger Christian sisters in Europe. The Guide du Voyageur is similarly adamant that Ethiopian Christians were more civilised than the Muslim or “pagan” inhabitants of the Ethiopian lowlands, many of whose lands had been incorporated in the Ethiopian state only few decades before in the wake of Menelik’s expansion- ist campaigns of the end of the nineteenth century.24 The establishment of this civilisational hier- archy between Christian and non-Christian Ethiopians capitalised on older prejudice, but should not be seen as merely the product of the cultural chauvinism of Ethiopian Christian elites. Once again, we should interpret the construction of this domestic hierarchy in the context of Ethiopia’s bid for sovereign statehood. Recent research on the Ottoman empire has shown that the ability to colonise and civilise other lands and people was regarded at the time as an import- ant legal evidence of “civilisation” and thus “sovereignty”.25 The Ethiopians did not have their colonial empire, but had their own internal others, racially and religiously different from their rul- ers, towards which the state could claim to be performing the same civilisational, developmental duties that the international community was performing towards the Ethiopian rulers. The Ethiopian government could be trusted not only with abiding by international law, but also enforcing its same principles on people constructed as “even-more-backwards”.26 2. The Japanese model in Ethiopian political thought Many other non-European polities were faced with the same dilemmas as Ethiopia, and were struggling to assert their sovereign statehood in a Eurocentric international system. Japan was one of them. Ethiopia turned to Japan and Japan turned to Ethiopia for the same reasons: to enhance their respective claims to sovereignty. Even at the peak of Ethio-Japanese relations in 6 S. MARZAGORA the early 1930s, though, theirs was always a hierarchical relationship, and the rest of the article will explore the nature and meaning of this hierarchy. The Japanese ruling elites, for once, had looked at Africa as a possible avenue of economic or territorial expansion. Just like other polities that had a hybrid status under international law – alternatively recognised as sovereign and non-sovereign, or defined as “semi-civilised” – Japan kept a close eye on European colonialism and variously pondered whether to join in the colonial expansionist momentum. Strong of its victory in the 1894-95 war against China, Japanese intel- lectuals and policymakers studied very carefully European colonial administration in Africa and Asia. The Japanese government’s believed that “a European-style empire [ … ] would earn Japan the respect of the Great Powers and eventually lead to the recognition of Japanese equality”.27 The colonisation of Korea was seen as an important step towards Japan’s assertion of full sover- eignty rights on the international stage. British colonial administration in Egypt was closely moni- tored and ended up being chosen as the model for Japan’s rule in Korea.28 The Japanese were ready to take advantage of any opportunities to increase their empire, including beyond East Asia.29 When it came to Ethiopia, for example, Japanese authorities con- sidered sending an officer, a doctor, and an accountant to join the Italian expeditionary force that invaded Ethiopia in 1895, as a possible gateway to a greater military and politico-economic presence in the African continent. Some members of the Japanese elite explicitly called for Japan to participate in the partition of the African continent. In an 1899 pamphlet titled Afurika no Zento (“The Future of Africa”), for example, a professor of law at the Tokyo Imperial University, Tomizu Hirondo, predicted that the “whites” would reap great economic benefits from the colonisation of Africa, and Japan would lose out if it did not join in. 2. The Japanese model in Ethiopian political thought Tomizu Hirondo’s reasoning makes again a clear connection between colonialism and sovereignty: if the “yellow” race did not work hard to catch up with the "whites", he states as a corollary to his call for a Japanese colonisation of Africa, they would end up being themselves “enslaved”. Tomizu was advocating some kind of sub-imperialism, or defensive colonisation, much like the attempt on the part of the Ottoman Empire to take part to the Scramble for Africa a couple of deca- des before.30 But if the Ottoman Empire had ultimately succumbed during the First World War, Japan was on the rise, and its imperial conquests in East Asia and the Pacific were expanding. This must have been a source of admiration for Ethiopian thinkers, who held the hope of one day bringing the whole of the Horn of Africa under Ethiopian control. To Ethiopian intellectuals, Japan repre- sented maybe the only example of a “small” nation that became a global power contender on par with Europe, while at the same time successfully defending a degree of “otherness”. This was not only the perception of Ethiopian thinkers, of course. Japan’s victory against Russia in 1905 had a profound impact on Asian and African intellectuals, and the decades that followed saw an outpouring of political pamphlets, newspaper articles, and treatises advocating the Japanese model in places like India, China, Vietnam, Egypt, Iran, and Turkey.31 We see here the core ten- sion that would shape Ethio-Japanese relations into the 1930s: while at least some members of the Japanese ruling elites looked at Africa as a possible target of imperial expansion, the Ethiopians saw Japan as a case study to buttress their own bid for independence. The first mention of Japan in Amharic print culture is perhaps a 1900 poem by G€abr€a-Ə gziabher Gila-Maryam (1860s-1914) which clearly frames modernity as a defensive endeavour: “He who accepts it, fears no one/He will become like Japan, strong in everything”.32 G€abr€a- Heyw€at Bayk€adan (1886-1919) concluded his 1912 At:e M@nil@k-nna Ityopya (“Emperor M@nil@k and Ethiopia”) with the advice to L@jj Iyasu, who had succeeded an ailing Mǝnilǝk on the throne, to follow the example of the Japanese government, especially when it came to educational pol- icy: “when the Japanese Government finds someone willing to go to Europe to learn, it supports them by giving them money. [ … ] As a result, the people (of Japan) opened their eyes. 2. The Japanese model in Ethiopian political thought They became rich, strong and respectable. [ … ] China and Asia have been following the path of Japan with great enthusiasm”.33 In a 1925 speech, W€arḳen€ah Ə s€ate (1865-1952) also argued that 7 THE INTERNATIONAL HISTORY REVIEW 7 7 education was the key to Japan’s success: “Realizing that to be successful in life they ought to imbibe European knowledge and imbibe it fast, [the Japanese] began to work diligently and were able to reach in sixty years the level of development that it has taken others centuries. [ … ] The reason behind the success of the Japanese to successfully defend their independence is their mastery of knowledge and education in due time. [ … ] Let us follow this amazing and praiseworthy example of far-sightedness and resoluteness of an entire people”.34 By that time, a consensus had arisen that Ethiopia was exactly sixty years behind Japan, calculated from the Meiji restoration of 1868. “Sixty years ago”, remarked Deressa Am€ante on a 1927 issue of the Amharic newspaper B@rhan@nna S€alam, “Japan was in the same state as Ethiopia”.35 G€abr€a-Ə gziabher Gila-Maryam, G€abr€a-Heyw€at Bayk€adan, W€arḳen€ah Ə s€ate, and Deressa Am€ante represented a new type of intellectual in Ethiopia: foreign-educated, well-travelled, glo- bally-oriented, and fluent in foreign languages. They were all in various ways involved with the imperial court and sought to influence policy-making. Bahru Zewde describes them as Ethiopia’s “pioneers of change” or “progressives”, in opposition to the “conservative” bloc. From their expe- riences abroad, these “progressive” intellectuals were all aware that foreign recognition had become as important for Ethiopian emperors as domestic recognition, and sought ways to reinforce the powers of the state in order to strengthen Ethiopia’s claim to sovereignty. They all saw the expansion of education as essential to create the new type of individual and collective subjectivity required by international law. Educational policy would be fundamental to create a secular political domain under the control of the Emperor and limit the powers of the Orthodox Church, which up until that moment had detained the monopoly over education and provided a powerful counterbalance to the power of the imperial crown. Not all of the “pioneers” embraced a rigidly unilinear conception of development, but the quantification of Ethiopia’s lag at sixty years must have been empowering in a moment when the future of Ethiopia’s independence seemed uncertain. 3. The 1931 Ethiopian constitution By the time he finally sat on the imperial throne in 1930, Haile Selassie could count on a good- sized group of “progressive” intellectuals. After the coronation, he moved to further centralise power into his own hands, and it is at this point that the Japanese model became operative. The 1931 Ethiopian constitution was closely modelled on the 1868 Meiji constitution, in terms of both content and division into articles.36 What is the significance of this choice? Ethiopia’s precarious international standing had not changed, and Haile Selassie’s priority was to reinforce Ethiopia’s claim to sovereignty. The pres- ence of modern legislation was again regarded as of the main attributes of a sovereign state. The more persuasively a state proved capable to enforce the rule of law on its territory, the more easily Europeans could be convinced to scrap unequal treaties and extraterritoriality clauses. Unequal treaties were “gradually abandoned as the Japanese, Chinese, Siamese and other ‘semi-sovereign’ states provided institutional proof of their capacity to ‘furnish’ Europeans (and, if necessary, their trading partners) with these rights independently – for instance, with the promulgation of the ‘Meiji Constitution’ in Japan. Only after these entities had ‘voluntarily’ inter- pellated as self-governing states of their own accord were they re-recognised, as it were, as ‘full’ (or fuller) subjects of international law”.37 The legal framework laid out by the 1868 Meiji consti- tution had allowed Japan to overwrite some of the unequal treaties that had previously regu- lated Japanese-Western relations. That the Ethiopian constitution served precisely this same purpose was transparently admitted by the intellectual who wrote the first draft of the document, T€akl€a-Hawaryat T€akl€a-Maryam (1884-1977). 2. The Japanese model in Ethiopian political thought The number could also be used to reassure Europeans by offering them a timeline for Ethiopia’s development “deliverables”, so to say. These intellectuals were aware that Ethiopia did not meet, at the time, the “standard of civilisation” and that the preservation of their country’s independence was premised on convincing European powers that Ethiopia had a stable government committed to fulfilling the international obligations of sovereign statehood. The adoption of the teleological historical framework of the "sixty years", therefore, was not a sign of cultural alienation, but the opposite – evidence of these intellectuals’ lucid understanding of their present moment, in which this teleology of progress, enshrined in international law, determined the recognition of both Ethiopia’s sovereignty and their own personal sovereignty as subjects. The Japanese case proved that it was possible to complete the developmental arc from “backward” to “advanced”, and to do in in a relatively short amount of time. The “sixty years” narrative, of course, implied that Japan had made it: in 1868, it started an irresistible process of modernisation that kept stably unfolding over the decades, turning Japan into a world power in just roughly half a century. At a first glance, such extensive discursive engagement with Japan on the part of Ethiopian intellectuals could be puzzling. Ethiopia’s claim to sovereignty and membership to the League of Nations revolved almost entirely around Christianity. Ethiopia was an ancient Christian monarchy and thus a natural member of the family of Christian nations. Japan was not Christian, so it might be unclear how references to Japan’s developmental trajectory might support the Ethiopian case. The answer is that the example of Japan complemented, rather than supporting, Ethiopia’s case. By stressing the Christian character of the Ethiopian monarchy and its commitment to civil- ise the non-Christians within its territory, Ethiopian ruling elites could claim sameness with Europe – the sameness on which political recognition and sovereign equality were predicated. Yet, civilisation was defined by European powers (and as a result also by international law) not only based on religion, but also based on race. Civilisation was Christian and white. On this latter criterion, the Ethiopians were destined to remain “other”, and thus to be permanently con- structed as uncivilised and unable to form an “effective” and “responsible” government. This is 8 S. MARZAGORA S. 2. The Japanese model in Ethiopian political thought MARZAGORA 8 where Japan was useful: not only was Japan a “small” state that had become “big”, but specific- ally a non-white state that had become “big” and gained full sovereign rights. The Yellow Peril discourse in the West implicitly highlighted the racial significance of Japan’s rise and the way it challenged the racist underpinnings of international legal categories. Japan, then, allowed the Ethiopians to claim that their racial “otherness” would not prevent them from rising to the level of European nations politically, economically, and militarily. Or, put differently, Japan was the proof that Ethiopia’s racial “otherness” did not condemn the country to be conquered and colon- ised. Stretched even more, Japan was seen as a guide on how to gain international acceptance while defending a degree of “otherness”, or better, while reserving the right not to entirely com- ply with the Western definition of civilisation. The 1931 Ethiopian constitution is a case in point. 3. The 1931 Ethiopian constitution In the speech he gave before all Ethiopian notables at the official promulgation of the constitution, he explained that “Ethiopia shall be protected and pacified by Law, which means, on the one hand, that she will improve her standard of living and grow wealthy, and on the other hand, that she will gain the respect of her neighbouring and other countries of the world, and benefit by concluding treaties in consonance with her ultimate interests”.38 The fact that the Emperor had “voluntarily” gifted a constitution to his people from above, and that the constitution came into effect “in a peaceful and orderly manner” was in itself “sufficient to gain for Ethiopia equal respect with other civilised people”.39 The Ethiopian people should thank the Emperor for this gift (“unparalleled in the history of the world”) because “we know that [the con- stitution] is not only for us”, but will also provide "for the safety of the independence of the future generations”.40 If the constitution strategically “interpellated” Ethiopia “as a self-governing state”, it did not do it along Western lines. In other words, the form of the constitution catered to the Eurocentric definition of statehood, but its content did not. The document approved in July 1931, as noted by virtually every historian, simply reconfirmed all the power hierarchies that already structured the Ethiopian state. In particular, the constitution insisted that the power of the Emperor was absolute, inviolable, and incontestable. T€akl€a-Hawaryat explains this choice in his speech before THE INTERNATIONAL HISTORY REVIEW 9 9 the Ethiopian nobility, where he lists four possible forms of government. The first three are decli- nations of a monarchical government with the king at the top: “tyrannical government”, “government by patience and customs”, and “united government acting according to the Law”. The fourth type is the “government of the people”, i.e. a democratic government, and T€akl€a- Hawaryat rules it out for Ethiopia in unambiguous terms: “such a government is not for unedu- cated and backward people; even amongst civilized people it continues to cause bloodshed”.41 This is an intriguing statement, as it seems to make two partly contradictory points. The first part of the sentence alludes at the usual unilinear teleology of development, and acknowledges that Ethiopia is still lagging behind. 3. The 1931 Ethiopian constitution The second part of the sentence seems to dismiss the demo- cratic form altogether, or at least expresses doubts over whether it is really the most advanced. The third form of government, what T€akl€a-Hawaryat calls “united government acting according to the Law”, meaning essentially a constitutional monarchy, is not only appropriate for Ethiopia in its current stage of development, but might be the best in absolute terms. T€akl€a-Hawaryat’s argument is cunningly crafted to rebut any possible Western objections to the authoritarian character of the constitution: firstly, he concedes to Western powers that Ethiopia is behind, and its uneducated citizens are not yet ready to participate in political decision-making. Then, he reminds Europeans that their democratic governments did not fare any better in terms of pre- venting violence and war. The last, unspoken weapon in his argumentative arsenal is precisely that the constitution is copied from the Japanese one: if the Meiji constitution worked for Japan to the point that Japan gained full international sovereignty, why couldn’t Ethiopia follow the same trajectory? In other words, the Meiji constitution allowed T€akl€a-Hawaryat to comply to the rules of the international society and partly resist them at the same time. It offered a blueprint for a form of sovereign statehood that was not completely predicated on the values of democracy that Western states ostensibly considered the hallmark of modernity from the Enlightenment onwards. It rather offered a modern way to legally codify the emperor’s sacredness and inviolability (chapter 1, article 3 of the Meiji constitution and chapter 1, article 5 of the Ethiopian constitution). In other words, it allowed the Ethiopian elites to fit into the existing international society and into its hegemonic ideology on unilinear progress, while at the same time defending a degree of “otherness” and preserve the political theology that for centuries had regulated power in the Empire. In this sense, the form of the constitution was designed to appease the international society, while its content was designed to appease the conservative power bloc at home. While this mediation was not always easy, the Ethiopian government did not want (nor it could afford) to stray from it. The years immediately after Haile Selassie’s coronation saw a diplo- matic and economic convergence between the Ethiopian and the Japanese state. 4. Economic and political relationships between the two governments From the point of view of the Ethiopian government, a rapprochement with Japan had a num- ber of advantages. Japan did not pose any direct territorial threat to Ethiopia, and an ally outside of Europe could come in handy if Britain, France or Italy started acting against Ethiopia’s inter- ests. Japan could also be a useful ally at the League of Nations, where the Ethiopian government feared isolation. But at a more basic level, every foreign alliance would boost Ethiopia’s state cre- dentials on the international scene and heighten Haile Selassie’s standing in the eyes of his sub- jects. The “capacity to enter into relations with the other states”, as the 1933 Montevideo convention would put it, was seen as a defining trait of sovereign statehood.44 Relationships with foreign powers could also serve the domestic power framework of the absolute monarchy. The American consul Addison Southard observed that the Ethiopians wanted a Japanese legation set up in Addis Ababa “to enhance the pride and prestige of their Emperor”. Haile Selassie knew that scenes of foreigners from powerful countries bowing to him would have a huge impact on Ethiopian public opinion. According to Southard, it was not difficult to imagine Haile Selassie’s officials “pridefully remarking, ‘See how even the great Emperor of Japan sends an important representative to bend the knee to our even greater Haile Selassie!’”.45 Besides the potential to boost Haile Selassie’s domestic and international reputation, a rap- prochement with Japan was also thought to offer economic benefits. Japanese products were much cheaper than Western ones: Japanese cotton cloth, for example, was eighty percent cheaper than American cloth.46 Already by 1918, Japanese cloth had superseded American mus- lin as the biggest textile import.47 This was a conspicuous slice of Ethiopia’s trade balance: in 1935 cotton cloth made up half of Ethiopia’s imports.48 Closer commercial relationships with Japan could also lessen Ethiopia’s economic dependency on Europe, although the Ethio- Japanese commercial relationship reproduced previous patterns of dependency: Ethiopia would export raw cotton and import cotton cloth. Despite the internal opposition of some prominent Ethiopian economists, G€abr€a-Heyw€at Bayk€adan above all,49 the Ethiopian government seemed prone to continue with the economic strategy to diffuse dependency in order to minimise its risks. Building an international reputation of good capitalist players, the Ethiopian government hoped, would bring benefits able to offset the possible dangers of asymmetrical commercial relationships. 3. The 1931 Ethiopian constitution Although it is tempting to see this convergence as a South-South alliance against the West, the relationship between the Ethiopian government and the Japanese government always kept to the spirit of international law. Neither the Ethiopian government nor the Japanese one intended to antagonise Western powers, and they both retraced their steps once Western states opposed Ethio-Japanese alliance. In fact, the alliance kept to the spirit of international law so much that it replicated the usual hierarchies of international relations. Japan was interested in Ethiopia as a source of raw materials and as a market for its manufactured goods. At least some representa- tives of the Japanese Ministry of Foreign Affairs held disparaging views of the Ethiopians as bar- baric, lazy, and uncultured.42 The members of the Japanese government did not “overly romanticize any presumed racial solidarity binding the Japanese with the Ethiopians”.43 This stood in contrast with some important sectors of Japanese civil society, as we shall see, to the point that Ethio-Japanese relations came to be precisely defined by two different worldmaking projects: the two governments’ and non-government actors’. 10 S. MARZAGORA 4. Economic and political relationships between the two governments The Japanese government maintained however a restrained approach to trade relationships with Ethiopia. In 1927, the two governments signed a (fairly standard for the time) treaty of friendship and commerce, but the Japanese government hesitated on whether to immediately set up a legation in Addis Ababa or delay the move until later. For Japan, Ethiopia offered the possibility to boost Japan’s manufacturing exports in a country that was not yet saturated with European goods, but the Japanese government was hesitant about the business opportunities offered by Ethiopia. A report on the state of the Ethiopian economy commissioned by the Japanese Ministry of Commerce did not yield encouraging results. The purchasing power in Ethiopia was extremely low,50 the cash economy very small, and transportation infrastructure for the most inadequate except for the French-owned Djibouti-Addis Ababa railway. The report rec- ommended to reinforce Japan’s position as main source of Ethiopia’s cloth imports by opening a direct maritime trade route between Japan and Ethiopia, thus bypassing the Indian merchants that the Japanese state had been relying on to deliver the products in the last leg of their Indian Ocean journey. The Japanese government, however, did not act on this recommendation. The diplomatic relationships between the two countries became closer in 1930-1. With the economic downturn of the Great Depression, Japan was now more interested in exploring com- mercial opportunities in Africa.51 Haile Selassie invited the Japanese government to his 1930 cor- onation ceremony, and the Japanese sent their ambassador in Turkey to attend the ceremony as Japan’s official delegate. To reciprocate the diplomatic visit, an Ethiopian delegation travelled to Japan in 1931, headed by the powerful Ethiopian Minister of Foreign Affairs H@ruy W€ald€a-Selasse 11 11 THE INTERNATIONAL HISTORY REVIEW 11 (1878-1938). The main objective of the visit was to develop closer commercial ties and stimulate Japanese investments in Ethiopia, but the Ethiopians covertly hoped to discuss an arms deal as well.52 The Ethiopian delegation received a very warm welcome, but the arms deal did not materialise. The Japanese government was, at the time, dealing with the international fallout of the Manchurian invasion, and was reluctant to get involved in any other potentially controversial military arrangement. In the forty days they spent in Japan, H@ruy and his party attended a num- ber of high-profile receptions and visited factories, offices, industrial farms, zoos, theatres, rail- ways, shrines, museums, and military training schools. 4. Economic and political relationships between the two governments As part of his visit, he met Emperor Hirohito, to whom H@ruy reportedly announced: Our Ethiopian Emperor is deeply impressed with Japanese Empire’s remarkable and great progress of the last sixty years, and is moved with surprise that the Japanese Empire accomplished such a great deed in such a short time. [ … ] He is determined to advocate to his whole nation to take the Great Japanese Empire as the best model.53 This visit marked the highest point of the diplomatic relationships between the two countries, although the Ethiopian government seemed keener than the Japanese one to tighten the alli- ance. H@ruy W€ald€a-Selasse was so impressed with his trip that, once back in Ethiopia, he promptly put together a booklet titled Mahd€ar€a B@rhan H€ag€ar Japan (“The Place of Light: the Country of Japan”, 1932). The book listed a number of historical similarities between Ethiopia and Japan, especially when it came to their monarchical history. The argument that the two countries had nearly identical historical trajectories allowed H@ruy to make a case for a closer political alliance. It also allowed him to make the usual argument that, given that the two coun- tries had always proceeded in history along parallel lines, Ethiopia would surely become like Japan in sixty years. Mahd€ar€a B@rhan H€ag€ar Japan was translated into English by Oreste Vaccari (1886-1980), an Italian linguist and Orientalist scholar, and then from English into Japanese by Oreste’s Japanese wife Enko (1896-1983). Much like Haile Selassie thought that a Japanese lega- tion in Addis Ababa would increase the prestige of the Ethiopian crown, the Japanese govern- ment proved keen to capitalise on H@ruy’s praise. The Japanese translation of Mahd€ar€a B@rhan H€ag€ar Japan was published in Tokyo in 1934 with a preface penned by the Minister of Foreign Affairs Kijuro Shidehara.54 Haile Selassie had been keen to show to his subjects how even Japanese appointees from the Far East bowed to his power, and the Japanese government was keen to show that even a remote country like Ethiopia was impressed with Japan’s greatness. 5. The role of non-government actors in Ethio-Japanese relations Some businessmen very lucidly noted that any Japanese commercial inroads would be perceived by the Italians, British and French as a threat to their interests, and recom- mended (against the report previously commissioned by the Japanese government) that the Japanese conduct all of their business via foreign intermediaries. The Japanese newspaper Osaka Mainichi sent to Ethiopia correspondents that were militant about tightening Ethio-Japanese relations. One of them, Yamauchi Masao, volunteered to person- ally negotiate with the Japanese government a loan for Ethiopia. He said he was disappointed that his government had turned down H@ruy’s request for a loan in 1931, and offered to lobby the Japanese government on behalf of Ethiopia to rectify the situation. Another correspondent of the Osaka Mainichi who felt that his government was not doing enough for Ethiopia was Shoji Yunosuke. Shoji and Yamauchi had too initially travelled to Ethiopia in search of business fortunes, but they also had a much larger political vision for Ethio-Japanese relations. They were both Pan-Asianists, and conceived of the Ethio-Japanese alliance as a racial coalition of non- white people against white supremacy. Shoji articulated this vision explicitly in his writings: Colored peoples in Asia and Africa have been suffering for a long time under white oppression. [ … ] The security and dangers facing Eastern peoples and the various peoples of Asia and North Africa are as the security and dangers for Japan itself. This concept is called Pan-Asianism. These oppressed peoples all view Japan as Asia’s leader, [and] friendship between Japan and Ethiopia has rapidly developed in such an atmosphere.56 This quote perfectly illustrates the constitutive ambivalence of Pan-Asianism. Japanese Pan- Asianists wrote powerful indictments of European colonialism, denouncing Western racism and calling for Asia’s liberation. The liberated Asia they envisioned, however, would be under Japanese leadership. Pan-Asianism, in this sense, “served not only as a basis for early efforts at regional integration in East Asia, but also as a cloak for expansionism and as a tool for legitimiz- ing Japanese hegemony and colonial rule”.57 In this sense, Pan-Asianism was, for a sector of the Japanese intelligentsia, a project of anti-Western sub-imperialism, and this explains why many Pan-Asianist organisations were also ultra-nationalist. As a consequence, Japanese Pan-Asianists talked about Asian freedom without problematising Japanese colonialism in Korea and China – a contradiction that undermined the Pan-Asian project and that caused profound rifts at the 1926 Pan-Asiatic conference in Nagasaki. 5. The role of non-government actors in Ethio-Japanese relations The Japanese government favoured diplomatic and commercial ties with Ethiopia, but was also reluctant to intensify them beyond a basic level of mutual cordiality. Some sectors of Japanese civil society thought otherwise. The economic relationships between the two countries were mostly driven by private businessmen, who, following H@ruy’s 1931 visit to Japan, started seeing Ethiopia as a potential new market as well as a land of untapped resources. Ethiopia’s develop- ment policy, as we have mentioned, mostly relied on concessions to foreigners for the building of infrastructure or the economic valoristion of the land. In the 1920s and early 1930s, a number of Japanese businessmen travelled to Ethiopia seeking concessions. These businessmen were very few in number, but their economic activities had a big impact on the international relations of the time. A case in point is Kitagawa Takashi, a Japanese merchant who travelled to Ethiopia in 1932 on behalf of the Nagasaki Association for Economic Investigation of Ethiopia (known as Nikkei-sha). Calvitt Clarke describes him as a “glib-talking and unscrupulous fixer”.55 His track record is indeed quite dubious. In Ethiopia, he was received by H@ruy W€ald€a-Selasse, to whom Kitagawa asked to lease more than 12,300,000 acres of land. Nikkei-sha would use the land to grow cotton, tobacco, tea, rice, wheat, fruit trees, vegetables, and medicinal plants, both for the Ethiopian market and for export. Kitagawa also asked for exclusive rights to grow opium – a risky 12 S. MARZAGORA 12 S. MARZAGORA 12 proposal, since the League of Nations was working to curb and control the opium trade. To manage the concession, Kitagawa envisioned an (overambitious) settlement scheme which would bring 650,000 Japanese to Ethiopia. The Ethiopian and the Japanese government seemed to ini- tially support Nikkei-sha’s plans, and they both provisionally authorised Nikkei-sha to grow medi- cinal plants. Kitagawa flaunted the concession as a done deal, although he had yet to sign any contract, and the news soon leaked to international newspapers, triggering a domino effect of exaggerated and sensationalist reports. Other Japanese businessmen took the initiative to explore Ethiopia’s economic potential, for example representatives of the Kanegafuchi Spinning Company and of the Chukyo Trading Company of Nagoya. Most of these initiatives came to nothing. Japanese businessmen soon found confirmation that purchasing power in Ethiopia was too low to sustain a Japanese import/ export business. 5. The role of non-government actors in Ethio-Japanese relations We can assume that the Pan-Asianists’ intentions towards Ethiopia were not different: the solidarity towards Ethiopia was probably conceived as a way to (at the very least) bring Ethiopia under Japanese control. Since the rise of Pan-Asianist ideas towards the end of the nineteenth century, Japanese Pan- Asianists always had a tense relationship with their government. They were frustrated by “the government’s realist course and its efforts at establishing a positive image of Japan in the West” to counteract the Yellow Peril fears.58 The Pan-Asianists were highly critical of their government’s capitalist internationalism and its political cooperation with Western countries. The rejection of Japan’s Racial Equality Proposal at the 1919 Paris Peace Conference further entrenched the Pan- Asianists’ resentment against the West and their disillusionment towards the League of Nations. THE INTERNATIONAL HISTORY REVIEW 13 The Yellow Peril narrative showed the extent of the discriminatory and pejorative views Westerners held towards Easterners. For th Pan-Asianists, these views should not be appeased or placated, but opposed in their entirety. For the Japanese government, the Pan-Asianists were a source of apprehension, as their vocal anti-Western ideas could undermine Japan’s relationships with Western powers. The Pan-Asianists’ “turn to Ethiopia” of the early 1930s generated a similar apprehension among the Japanese government. The growing number of pro-Ethiopian publica- tions of the 1930s and the way they captured the imagination of the Japanese reading public soon started interfering with Japan’s foreign policy. This situation had a striking parallel in Ethiopia, where a new generation of foreign-educated Ethiopians, nicknamed the “Young Ethiopians” by foreign commentators, was becoming increas- ingly critical of Western racism, causing more than a diplomatic headache for the Ethiopian gov- ernment. The Young Ethiopians were distinct from the “progressives”, who were generally older and occupied higher positions of power and prestige. Many Young Ethiopians worked in the public administration, but their relationship with the imperial bureaucracy was strained. “Most foreign commentators", observes Pankhurst, “differentiated between the Young Ethiopians and the Ethiopian Government – and in some cases even regarded the former as the critics of latter”.59 The Young Ethiopians’ hostility towards outsiders was notorious, and foreign observers, no doubt offended in their feelings of racial superiority, often accused them of “xenophobia”. Two successive British Ministers to Ethiopia, first Charles Bentinck in 1927 and then Sir Sydney Barton in 1928-29, repeat the accusation. 5. The role of non-government actors in Ethio-Japanese relations Both were condescendingly unsympathetic towards the Ethiopian people, and they felt outraged that the Ethiopians were not inclined to conform to their idea of racial hierarchies. Bentinck complained of the situation in the following terms: As in China and India, so in Abyssinia, there is a small section of the younger generation which has received a smattering of Western education. These young men are satisfied with having scratched the surface and think they know enough. In the town of Addis Ababa they consort in terms of equality with the riff-raff of Armenia and Greece, and in some cases France and Russia. They get the idea that they are not only the equal, but the superior of the white man, and they strive to show this in various forms: refusing to pay salaries due to Europeans for services rendered, and by throwing them penniless into the streets or using personal violence against their persons and properties, etc.60 No doubt these were precisely the racist attitudes that the Young Ethiopians resented. The colonial machinations of European powers towards Ethiopia further incensed them. The December 1925 agreement between the Italians and the British, whereby the British recognised Italy’s exclusive economic influence in Western Ethiopia and Italy supported the British project to build a dam on Lake Tana in Northern Ethiopia, spurred waves of turmoil in Addis Ababa. The Young Ethiopians were reportedly at the forefront the protests. According to British traveller Charles Rey, “foreigners were stopped in their cars and allowed to drive on only if they were cer- tified to be neither British nor Italian”.61 A similar instance of unrest happened in 1933, when several Young Ethiopians were thrown out of a Greek coffee shop, the Tabaris Cafe. The motivations of the cafe owner are unclear, but the expelled evidently felt they were being discriminated against. They returned six days later to smash up the place in reprisal, causing the intervention of the police and the closure of the cafe. In the incident were reportedly involved all the major figures associated to the Young Ethiopians: Kidan€a-Maryam Ab€arra, F€ak€ad€a-Sellase H@ruy, W€ald€a-Giyorgis W€ald€a-Yohann@s, M€akonnen Habt€a-W€ald, B€asahwer€ad H€abt€a-W€ald, and Ayy€al€a G€abr€a. The terrain was fertile for the Japanese Pan-Asian arguments to take hold among this youth, and the Ethiopian government must have been worried about an anti-Western, anti-white ideological escalation. More individual initiatives jeopardised the Ethiopian government’s and the Japanese govern- ment’s relations with Europe. 5. The role of non-government actors in Ethio-Japanese relations When Shoji Yunosuke left Ethiopia in 1933, Haile Selassie entrusted him with delivering a gift to Sumioka Tomoyoshi, who had hosted the 1931 Ethiopian delegation headed by H@ruy W€ald€a-Selasse. Sumioka was, like Shoji, a pro-Ethiopian Japanese nationalist and Pan-Asianist. He appears in a now-famous photo that portrays the four members of the 14 S. MARZAGORA S. MARZAGORA Ethiopian delegation sitting on their knees (except for H@ruy, who is seemingly sitting on a low stool) in the front row, all wearing Japanese kimonos. Sumioka and his wife are standing behind them in the back, Sumioka also wearing the same type of kimono as the Ethiopians, while his wife is wearing a traditional Ethiopian white cotton robe called net:ela. The solidarity between the two people is symbolically represented by the swapping of traditional clothes: Sumioka’s Japanese wife is wearing Ethiopia’s national dress, while the four Ethiopian men are wearing Japan’s national dress. The Ethiopian on the right end of the front row was Lǝjj (a title for young noblemen) Araya Abebe. Araya seems to have desired to marry a Japanese woman even before his 1931 grand tour of Japan, but the 1931 trip must have strengthened his resolve. At first, Haile Selassie and H@ruy W€ald€a-Selasse did not oppose the plan, and H@ruy cautiously asked Sumioka if he could help broker the marriage. Sumioka was enthusiastic at the idea. g In June 1933, Japanese newspapers published the announcement that the “immensely popu- lar” Ethiopian “prince” (vastly inflated credentials in both cases) that had visited Japan a year and a half before was looking for a Japanese wife. Sumioka sent H@ruy a shortlist of the best applicants and when the final decision was made in Addis Ababa in January 1934, H@ruy sum- moned Yamanouchi to the imperial palace to cable the news back to Sumioka in Japan. The chosen candidate, Kuroda Masako, came from a well-off aristocratic family, but had also received a modern education, was fluent in English, and fond of sports. The Japanese press was soon inundated by romanticised reports celebrating the fairy tale of the impending “royal wedding”. The press affiliated to Pan-Asian associations, on its part, aggressively promoted the wedding as a step towards the urgent alliance of non-white people against white colonialism. Kuroda herself seems to have operated precisely within the hierarchical logic of Japan’s Pan-Asianists. 5. The role of non-government actors in Ethio-Japanese relations She “saw herself as the first of many who would emigrate to Ethiopia”, explaining that “with its ever- increasing population, Japan would have to found colonies abroad”.62 The peak of the marriage preparations coincided with the peak of the media frenzy about Nikkei-sha’s alleged deal with the Ethiopian government. Japan’s political and economic pres- ence in Ethiopia seemed to have widened. This was mostly due to the private initiative of con- cession hunters and ideologically militant Pan-Asianist activists, with a certain degree of synergy between the two, for example about the possibility of a large Japanese population transfer. On the Ethiopian side the situation was similar: the marriage idea was a private initiative of Araya himself, and while H@ruy and Haile Selassie initially gave the young bachelor their blessing, the marriage was not something that H@ruy and Haile Selassie had themselves sought to arrange in the first place. The Japanese Pan-Asianists and the Young Ethiopians had similar ideas about how their respective governments should position themselves globally. They both perceptively understood that the universalism professed by international law and international organisation was a fac¸ade to defend Western colonial interests. The international society promised acceptance and recogni- tion to all those people that met the legal requirements of statehood and civilisation, without actually intending to grant any to non-white, non-European “others”. The compliance with those requirements, if anything, would make non-European states more dependent on Western diplo- matic endorsement and more vulnerable to Western economic penetration. The international space was constituted according to racist principles that made any Western talk of sovereign equality empty. In the global system of the time, Japan and Ethiopia would always be in a subor- dinate position – international law was designed for this very purpose. Why trying to constantly appease the Westerners, considering the dehumanising, discriminatory views they held of non- white, non-Christian, non-European people? As we have seen, however, the political vision of Japanese Pan-Asianists was not a remaking of the world under more egalitarian principles of horizontal solidarity. It was rather the creation of an alternative world-system in Asia, in which Japan could rise to prominence without any Western interference. The relationship with Ethiopia was probably understood in a similarly hierarchical way. It is not that genuine solidarity was THE INTERNATIONAL HISTORY REVIEW 15 15 15 absent, but Japan was always nationalistically presented as the alternative solution to a Western- dominated world. 5. The role of non-government actors in Ethio-Japanese relations On their part, the Young Ethiopians repeated some of the arguments of the “conservative” power bloc – maybe paradoxically, considering that in terms of their foreign education and glo- bal outlook they could not be more different from the far less travelled and less cosmopolitan aristocrats and church authorities that made up the bulk of the conservative faction. The two groups converged on their disapproval of Haile Selassie’s policy of integrating Ethiopia in the world-system as fast as possible in order to protect its sovereignty. The expansionist ambitions of European powers were in front of everyone’s eyes to see, making it clear that foreigners could not be trusted. The Europeans had abundantly demonstrated their disregard for Ethiopian tradi- tions, so how could the government be sure that they would suddenly change their mind if Ethiopia managed to develop its economy and infrastructures? Playing by the rules of those who wanted to conquer Ethiopia and undermine its cultural traditions would put the country even more in danger. Hence, the conservatives pushed for isolationism as a solution: Ethiopia had to go its own way, close its borders, refuse to integrate in the world-system. While the conserva- tives were reluctant to compromise on Ethiopia’s political identity, the Young Ethiopians were instead keen to see a transformed and modernised Ethiopia. Just, Ethiopia did not need foreign- ers to develop: it had all of the resources (including the human resources, and here the Young Ethiopians referred to themselves) for a self-driven development, without concessions to foreign- ers, foreign advisors, and foreign businessmen, who only worked to advance the interests and colonial schemes of their own countries. While the connection drawn between political and eco- nomic sovereignty was cogent, the Young Ethiopians’ argument that Ethiopia could modernise in isolation stood in contrast with their view of modernity as a project of global politico-eco- nomic integration. . The Young Ethiopians were right, though, that this project was a project of domination of a restricted group of countries over the rest of the world, and that Ethiopia would ultimately lose out in the process. 6. Government vs. non-government actors: The denouement of Ethio- Japanese relations These ideas were dangerous for the Ethiopian and the Japanese government, whose objective remained to fit into the international system and maintain good relationships with Western powers, while also trying to resist as much as possible a full assimilation into the civilisational ideals promoted by Western nations. The tension between these two different visions of world- making came to the fore almost immediately. As some Japanese had feared, Britain, France, and above all Italy immediately felt threatened by Japan’s diplomatic and commercial inroads in Ethiopia. Starting from September 1933, European newspapers reported on Nikkei-sha’s alleged deal with the Ethiopian government in exaggerated terms, inflating the scale of the concession and framing it as a premeditated chal- lenge to British, French and Italian interests in the area. News of an impending Ethio-Japanese arms deal further amplified this narrative. The European reports about the engagement between Araya and Kuroda in early 1934 were similarly sensationalist. Like in the romanticised articles published by the Japanese press, European newspapers talked about a “royal wedding” between the Japanese and the Ethiopian imperial dynasties. London was alarmed at the rumours that the Japanese intended to run the cotton trade on their own, without anymore relying on Indian commercial mediation. Rome, whose colonial ambitions towards Ethiopia were increasingly trans- parent, had an even stronger reaction. Italy knew that the rumours were largely false, but will- ingly amplified them, in an attempt to justify its looming invasion of Ethiopia as a defence of European interests against the Yellow Peril.63 16 S. MARZAGORA 16 S. MARZAGORA The press coverage of Ethio-Japanese ties was for the most part overblown. Araya and Kuroda came from the upper echelons of their respective societies, but were far from a “prince” and “princess” in their own imperial houses. Araya, for once, was only remotely related to the Emperor and was an overall unimportant figure in political and dynastic terms. Rumours about an alleged Japanese commercial takeover of Ethiopia were also hyperbolic. The concession to Nikkei-sha had never been finalised, and (to Ethiopia’s disappointment) no Ethio-Japanese arms deal had ever been signed. Commenting on the international controversy, Addison Southard reported that there was only a dozen of Japanese in the whole of Ethiopia, and most Japanese business ventures in Ethiopia had failed. 6. Government vs. non-government actors: The denouement of Ethio- Japanese relations Although the Ethiopian government had cunningly advertised the country’s business potential, in reality there was little capacity, in terms of resour- ces and infrastructure, to seriously incentivise any foreign investment.64 Accused of turning against European powers and fearing serious repercussions, the Ethiopian government and the Japanese government rushed to shut down the rumours. Neither had the least intention to compromise their relationship with the Great Powers. H@ruy explained to inter- national correspondents that he had indeed held preliminary talks with private Japanese individ- uals about a possible land concession, but that the deal was never finalised. He maintained that Ethiopia was interested in doing business with Japan while also maintaining cordial relationships with Europe. The Ethiopian government also backtracked on the marriage plans, withdrawing their support. The Japanese government intervened heavy-handedly to investigate matters. They were in a much more difficult position than the Ethiopian government, since Japanese public opinion was resolutely pro-Ethiopia and Japan’s Pan-Asian organisations were escalating their pro-Ethiopia campaign. Italy’s military support for China against the Japanese in 1932 had further turned Japanese public opinion against Rome. The Japanese government “did not agree with either Yamanouchi or Shoji and certainly took little but embarrassed notice of Kuroda’s hopes. Tokyo could not allow a free hand in Ethiopia to ambitious, pan-Asiatic adventurers such as Kitagawa—or for that matter Yamanouchi himself”.65 The government proactively tried to reassure Italy of its peaceful intentions, denying all the rumours of Japanese military aid to Ethiopia. The hierarchical nature of the relationship between Japan and Ethiopia quickly became appar- ent: Japan did not need Ethiopia as much as Ethiopia needed Japan. The Japanese ambassador in Rome, Yotaro Sugimura, went as far as precociously announcing in July 1935 that Japan would remain neutral in Italy’s upcoming war on Ethiopia. More tensions followed, but by the summer of 1935, Rome and Tokyo started reconciling and the press frenzy started subsiding. Italy’s strat- egy to paint Japan as a two-faced enemy who was secretly scheming against the West, though, had worked. Britain and France acquiesced to Mussolini’s attack on Ethiopia, among other rea- sons, also because their fears of Japanese meddling with European colonial affairs had been rekindled. The Italian narrative also contributed to persuading the Soviet Union, who had a his- tory of military tensions with Japan, to abandon its anti-imperialist policy and take a pro-Italian position at the League of Nations. 6. Government vs. non-government actors: The denouement of Ethio- Japanese relations The Ethiopian government, though, had not completely given up on Japan as a possible ally – or at the very least a possible source of military aid in the brewing conflict with Italy. Af€aw€arḳ G€abr€a-Iyy€asus, who in the meantime had been appointed Ethiopian ambassador to Italy, asked Sugimura for military help, which Sugimura refused. The last Ethiopian bid to revive the relation- ship with Japan was entrusted to Daba Birru, who had accompanied H@ruy to Japan in 1931 and can be seen on the left in the famous photo with Sumioka Tomoyoshi. In September 1935, when the Italian invasion was inevitable, and faced with the disappointing inaction of the League of Nations, Daba travelled to Japan in search of help. Daba’s mission was at the same time a public relations success and a diplomatic failure. Shoji accompanied Daba on the trip and his newspaper Osaka Mainichi gave ample coverage to Daba’s visit. Daba was hosted as a guest of honour by all those ultra-nationalist and Pan-Asianist organisations that had embraced the Ethiopian cause. The Black Dragon Society had organised THE INTERNATIONAL HISTORY REVIEW 17 17 17 an Ethiopia Crisis Committee, many existing associations (including women’s and youth associa- tions) took a firm pro-Ethiopia stance, and many new organisations were set up to campaign for Ethiopia. All these organisations scrambled to host Daba as their guest of honour, gathering large crowds to welcome Daba in each leg of his tour and to attend each of Daba’s public speeches. All the major exponents of Japan’s ultra-nationalist and Pan-Asianist political wings were involved, including Toyama Mitsuru, the founder of the Black Dragon Society and of the Dark Ocean Society. Influential Japanese, though, were largely absent, and Daba’s talks with offi- cials from the Japanese Ministry of Foreign Affairs came to nothing. The Japanese government was firmly committed to its policy of non-interference in Ethio-Italian tensions. The complete denouement of the Ethio-Japanese alliance was sealed by the mutual recognition of December 1936, when Italy recognised Japanese rule in Manchukuo and Japan recognised Italian rule in Ethiopia. The Ethiopian government’s policy to play Britain, France and Italy off against a fourth power could only work so far. The Italian justification for the invasion before the League of Nations was what the Ethiopians had always feared. 6. Government vs. non-government actors: The denouement of Ethio- Japanese relations Italy claimed that Ethiopia did not meet the criteria for sovereign statehood and that it was a tyrannical regime whose lowlands, far from being “civilised” its lowlands, had actually seen a recrudescence of the slave trade. This, Italy argued, was evidence that Ethiopia had not met the “special obligations” on which its membership (and its sovereignty) were condi- tional. Parfitt has shown that, from the point of view of how international law was constituted at the time, these Italian claims were not legally false, precisely because the “special obligations” had constructed Ethiopia as simultaneously sovereign and non-sovereign and premised its mem- bership on meeting of specific civilisational requirements, such as the eradication of the slave trade. As a consequence, we should not interpret the Abyssinian crisis as the failure of the League to apply international law, but rather as evidence that the problem was in the way inter- national law was itself constituted. From this point of view, the Young Ethiopians and the Japanese Pan-Asianists had lucidly captured that, structurally, the international system was never, and could never be equal. The issue was not the application of the law – the issue was that the law was designed from the start to reproduce that politico-economic inequality between the centres and peripheries of capitalism.66 7. Conclusions: “a politics of emulation”? There were many layers to the intensification of Ethio-Japanese relations in the interwar period. Ethiopian elites could use the Japanese case to challenge the racist ideology that underpinned the international system. Against that white supremacist ideology, Japan proved that non-white people were perfectly able to develop and modernise without the colonial intervention of the West. The Ethiopian government might have been a racial “other”, but was perfectly able to lead the country on the path to progress. This argument complemented the Ethiopians’ main bid for sovereignty, which claimed sameness with Europe on religious grounds and cast the Ethiopian government as the civiliser of its non-Christian lowlands. Overall, the Ethiopian government strove to abide by the rules of the international system, while having to defend some of its pre- vious political identity in order to placate some powerful conservative domestic factions. Japan was seen as a model for this type of mediation between domestic and international political cul- ture. The Ethiopian convergence towards Japan should be seen as a way to assert a more authoritative juridical personhood within the existing international system. The Japanese government, like the Ethiopians, also chose a policy of alliances with Western powers and, initially, of participation in international organisations such as the League of Nations. The intensification of Ethio-Japanese diplomatic relationships in 1930-31 was not, in any way, intended to challenge the international law of the time, but on the contrary to boost the two country’s sovereignty rights – in the case of Japan, also through a (very hypothetical and 18 S. MARZAGORA S. MARZAGORA 18 18 never materialised) colonial takeover of Ethiopia. In this sense, the relationship between the two countries was always hierarchical, and reproduced the same pattern of economic dependency that Ethiopia had with Western states. The power relation between the two countries became apparent when Japan chose to side with Mussolini over Haile Selassie when Italy invaded Ethiopia in 1935. The Italian justification for the invasion hinged precisely on Ethiopia not meet- ing the parameters of sovereign statehood, including failing to civilise its ethnically more “backwards” lowlands. Both Ethiopia and Japan had militant social groups that opposed their government’s pro- Western policies, and called for a more confrontational foreign policy against the racism that underscored Western diplomacy and economic policies. Both the Japanese Pan-Asianists and the Young Ethiopians opposed their government’s capitalist internationalism and conciliatory response to Western discriminatory attitudes. Notes (There are no surnames in Ethiopia, so Ethiopian authors have been referred to and listed by their first name fol- lowed by their father’s name, as common practice in Ethiopian studies). 7. Conclusions: “a politics of emulation”? These were the two factions that pushed more aggressively for a tightening of Ethio-Japanese relations – and between the two, the Japanese Pan-Asianists were particularly militant in their campaign. The way in which Japanese Pan- Asianists proactively tried to forge closer politico-economic relationships between Japan and Ethiopia in the interwar years is evidence of their expansive understanding of Pan-Asianism. Ethiopia was seen as potentially part of the Pan-Asian project, as some kind of “Orient writ large”. Despite their vastly different social positions, the Young Ethiopians shared many of the fears of the conservative power bloc. They thought that opening Ethiopia to Westerners would increase, not decrease, the risk of being conquered, and that Westerners could not be trusted to respect Ethiopia’s independence. This was a long-sighted assessment. Japan’s Pan-Asianists and the Young Ethiopians had intended to come together precisely to oppose European colonialism and white supremacy, and the Europeans reacted precisely by reasserting European colonial pre- rogatives and white supremacy. Reducing these complex power relations to a “politics of emulation” is a simplification to put it mildly. Calls for Ethiopia to become “more like Japan” were fewer in number than calls for Ethiopia to become “more like Europe”, and in both cases, these were sophisticated ideological pitches in support of a fragile claim to sovereignty. The conservatives’ position should not be seen as a patriotic defence of “tradition” and, equally, the progressives’ concessions to the inter- national system should not be seen as an intellectually-flawed betrayal of that “tradition”. The two blocs had different assessments of the workings of the international system and of the best strategy to protect Ethiopia’s independence, and argued for different solutions on how to medi- ate domestic and international political cultures. In other words, calls to “emulate” others were not for the sake of emulation per se, and should not be taken as evidence of a derivative, uncreative political thought. On the opposite, they were sophisticated responses to an inter- national system designed to reproduce Eurocentric politico-economic hierarchies, and thus stacked against Ethiopia. We are bound to misrepresent political thought if we analyse it as detached from its material conditions. (There are no surnames in Ethiopia, so Ethiopian authors have been referred to and listed by their first name fol- lowed by their father’s name, as common practice in Ethiopian studies). Notes on contributor Sara Marzagora is an intellectual and literary historian of Ethiopia and the Horn of Africa. She is currently a Lecturer (Assistant Professor) of Comparative Literature at King’s College London. Disclosure statement No potential conflict of interest was reported by the authors. THE INTERNATIONAL HISTORY REVIEW 19 THE INTERNATIONAL HISTORY REVIEW 19 ORCID Sara Marzagora http://orcid.org/0000-0003-3989-3080	5,150
https://openalex.org/W4288041843_1	Spanish-Science-Pile	Open Science	Various open science	2,022	Novel data-driven control approaches : application in essential oil extraction processes	None	English	Spoken	7,930	10,295	PONTIFICIA UNIVERSIDAD JAVERIANA DOCTORAL THESIS NOVEL DATA-DRIVEN CONTROL APPROACHES Application in essential oil extraction processes Author: Advisor: Eng. Freddy F. Valderrama Gutiérrez, M.Sc Prof. Fredy O. Ruiz, Ph.D. A thesis submitted in fulfillment of the requirements for the degree of Ph.D. in Engineering in the Departamento de Electrónica Facultad de ingenierı́a Pontificia Universidad Javeriana December 12, 2019 Abstract In this dissertation are developed new methods for the design of controllers from data for linear systems, useful when the plant model is unavailable, two frameworks have been considered using set-membership techniques (SMT) and the Youla-Kucera parametrization (YK-DDC). The developed methods have been extensively evaluated in simulation and experimental settings. In SMT, the Unknown But Bounded (UBB) noise structure affecting the experimental data has been assumed, and a proper data-based criterion is stated according to the reference model criterion problem. Such formulation allows to state hard bounds on controller parameters via linear programming techniques. SMT approach has been experimentally validated on an active suspension system via a Monte-Carlo experiment, and it has been compared with VRFT methodology. Results have shown that using the same information about the plant to be controlled, the VRFT controllers are strongly affected by the size of the dataset while the SMT controllers exhibit good performances, even when they are estimated from reduced datasets. Two SMT extensions have been proposed, the first one allows to design limited-complexity controllers using sparse identification methods, the proposed algorithm avoids solving big combinatorial problems that arise when the dimension of the vector parametrizing the candidate controllers is large and the number of desired parameters is much lower. A benchmark flexible transmission model is employed to illustrate the performance of the proposed methodology in comparison to the sparse correlation based tuning approach (SCbT). It is found that both approaches offer similar performance when the size of the data set is much larger than the dimension of the controller parameters vector. Notwithstanding, the SCbT controllers are strongly affected when data set size is reduced, while SMT controllers exhibit good performances even when the controller parameters are estimated from reduced data sets. The second extension proposes a procedure to estimate controllers capable of approaching a given closed-loop reference model and a sensitivity transfer function (Two-degree-of-freedom controllers). To do this, an efficient solution based on convex optimization is proposed. In literature, this the first attempt that employs a SM formulation. In order to validate our approach, the Errors-in-Variables approach proposed in literature for one DoF controllers has been adapted to solve the two degree-of-freedom problem. Our method and the adapted method are developed within a Set Membership framework. Results indicate that the computational cost, in terms of execution time of the adapted approach is higher than in our setting, even three hundred times bigger, while similar estimation errors are presented in both approaches. The Youla-Kucera parametrization has been employed to solve the problem of controllers design without requiring a process model. The main contribution of this part of the thesis is that the proposed controller structure allows reaching more stringent reference models than those proposed previously in the literature, maintaining a convex formulation and a procedure to estimate the closed-loop stability. An extension of YK-DDC methodology to the MIMO case is also proposed. Such a methodology has been experimentally validated on a 2-DoF Helicopter. This is an important model from the control engineering point of view due to its wide non-linear characteristics, highly cross-coupling effects, and instability in open-loop. Experimental results showed that our approach achieves better results compared with the LQR+I controller proposed by the system manufacturer since the former meets the control requirement with lower settling time and smaller maximum overshoot. However, given that the 2DOF helicopter is unstable an initial stabilizing controller is required to collect the dataset and to construct a cascade control strategy. The modeling and control of essential oil extraction processes are comprehensively studied, and novel strategies for optimal extraction operation and temperature regulation are proposed. A lab-scale ohmic distiller able to extract the essential oil from 100[g] of vegetal mass (aromatic herb) has been designed and constructed by the author. The selected characteristics of the distiller allowed to obtain measurable essential oil quantity from Eucalyptus, in order to obtain the kinetic extraction curves. In steam distillation, an optimal control problem has been formulated and solved in order to save energy during the extraction process while maintaining the yield of extraction. Our solutions show that optimal steam flow trajectories are not necessarily constant, as previously mentioned in the literature. Given some phenomenological similarities between steam distillation and Ohmic assisted hydrodistillation (OAHD), the optimal input trajectories found to the former were extended to OAHD. Experimental results indicate that our input trajectory allows important energy savings, maintaining the yield statistically equal to the yield with a constant input. In addition, the proposed YK-DDC approach is applied for the regulation of temperature in an ohmic-heater, where experimental results show that our controller offers better tracking results than a procedure extracted from recent literature. Acknowledgement This project was supported by Gobernación de Boyacá (Colombia), convocatoria 733 de 2015. Contents 1 Introduction 10 2 A set-membership approach to DDC 2.1 Statement of the problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.2 Set-Membership approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.3 Active suspension Tuning Case Study . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.3.1 Controller tuning problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.3.2 Monte-Carlo experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.3.3 Process disturbance experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.4 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 15 16 17 21 21 23 25 29 3 Limited-complexity Controller Tuning 3.1 Sparse controller tuning problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.2 A sparse Set Membership framework for controller tuning . . . . . . . . . . . . . . . . .. 3.2.1 The Feasible Parameters Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.2.2 Finding a sparse controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.3 Numerical Case Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.3.1 Monte-Carlo test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.4 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 30 31 32 35 36 38 41 41 4 2 DoF controller structures 4.1 Problem formulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.2 Sensitivity tuning via Set-membership . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.3 Closed-loop reference model tuning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.4 Illustrative examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.4.1 Comparison Set-membership approaches . . . . . . . . . . . . . . . . . . . . . . .. 4.4.1.1 Computational solution . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.4.1.2 Result analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.4.2 Comparison against VRFT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 4.5 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 43 43 45 47 48 48 49 49 51 52 5 A Youla-Kucera Parametrization for Data-driven Controllers Tuning 5.1 Statement of the problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.2 A stabilizing controller structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.2.1 A structure for Q. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.2.2 The Q filter in terms of data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.3 Q̂ tuning scheme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.3.1 Correlation approach to tune Q̂ . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.3.2 Selecting a proper number of parameters . . . . . . . . . . . . . . . . . . . . . . .. 5.3.3 Procedure to tune Q̂ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 54 55 55 56 56 57 57 58 59 4 5.3.4 Stability margin estimation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Numerical examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.4.1 Flexible transmission . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.4.2 Non-minimum phase plant equipped with integrator . . . . . . . . . . . . . . . .. Extension to MIMO case . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.5.1 Q̂ tuning scheme in the MIMO case . . . . . . . . . . . . . . . . . . . . . . . . .. 5.5.2 2-DOF helicopter case study . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.5.2.1 Controller tuning problem . . . . . . . . . . . . . . . . . . . . . . . . .. 5.5.2.2 Experiment design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5.5.2.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 60 60 60 65 66 68 68 69 69 70 72 6 Application in essential oil extraction processes 6.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2 An optimal control approach to steam distillation of essential oils from aromatic plants .. 6.2.1 Extraction process model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.1.1 Process description . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.1.2 Model description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.1.3 Model analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.2 Optimal oil extraction control problem . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.2.1 Optimal control problem . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.3 Results & discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.3.1 Case I: Maximizing yield . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.3.2 Case II: Minimizing energy . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.3.3 Case III: Trade-off between yield and energy . . . . . . . . . . . . . . .. 6.2.3.4 Sensitivity analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2.3.5 Applicability discussion . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3 Ohmic-assisted hydrodistillation (OAHD) . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3.1 Process description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3.2 Ohmic heating distiller design . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3.2.1 Camera design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3.2.2 Condenser designing . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4 Experimental tests for input power trajectories in OAHD process . . . . . . . . . . . . .. 6.4.1 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.1.1 Vegetal mass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.1.2 The mixture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.1.3 Experimental setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.1.4 Experiment description . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.2 Experiments performed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4.2.1 Scenario I: Constant input power trajectory . . . . . . . . . . . . . . .. 6.4.2.2 Scenario II: Decreasing input power trajectory . . . . . . . . . . . . . .. 6.4.3 Analysis and comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.5 Controlling the heating curve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.5.1 Controller design tuning problem . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.5.2 Tuning procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.6 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 73 73 74 74 74 74 76 77 77 78 78 78 79 80 82 82 82 83 84 85 86 87 87 88 88 88 89 89 89 91 92 92 93 95 7 Conclusions 97 5.4 5.5 5.6 5 A Annexes 99 A.1 Annex I: VRFT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 A.2 Annex II: sparse-CbT algorithm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 A.3 Annex III: Errors in Variables to 2DoF controller tuning problem . . . . . . . . . . . . . . 101 6 List of Tables 2.1 2.2 2.3 Active suspension parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Performance results of Monte-Carlo experiment . . . . . . . . . . . . . . . . . . . . . . .. Performance results of process disturbance experiment . . . . . . . . . . . . . . . . . . .. 22 27 29 3.1 3.2 3.3 SCbT convergence results for λ = 0.01 . . . . . . . . . . . . . . . . . . . . . . . . . . . .. SCbT convergence results for λ = 0.12 . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Results for Monte-Carlo experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 40 40 41 4.1 4.2 4.3 Parameters estimated via both approaches for SN R = 20dB . . . . . . . . . . . . . . . .. Parameters estimated via both approaches for SN R = 30dB . . . . . . . . . . . . . . . .. Average computation time for both approaches . . . . . . . . . . . . . . . . . . . . . . .. 50 50 50 5.1 5.2 5.3 Stability results for infinity norm criterion for both approaches. . . . . . . . . . . . . . .. Quality control results for both approaches in both cases. . . . . . . . . . . . . . . . . .. Results for YK-DDC and LQR+I controller in the case of 2DOF helicopter . . . . . . .. 63 64 71 6.1 6.2 6.3 6.4 6.5 6.6 Model parameters and initial conditions . . . . . . . . . . . . . . . . . . . . . . . . . . .. Results for different γ values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Results for different weighting factors . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Sensitivity analysis (γ = 0.02): Effect on saved energy . . . . . . . . . . . . . . . . . . .. Energy consumption in two different trajectories . . . . . . . . . . . . . . . . . . . . . .. Yield obtained in the experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 77 81 81 82 91 92 7 List of Figures 1.1 Thesis organization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 12 2.1 Assumed feedback control structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.2 Tuning problem block diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.3 Active suspension platform . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.4 Active suspension model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.5 Open-loop step response of the active suspension. . . . . . . . . . . . . . . . . . . . . . .. 2.6 Active suspension nominal frequency response. . . . . . . . . . . . . . . . . . . . . . . .. 2.7 Step response of Reference models M (q −1 ) for different α values. . . . . . . . . . . . . .. 2.8 Sample of suspension travel with added output noise. . . . . . . . . . . . . . . . . . . . .. 2.9 Controller structure selection results. N=1000 . . . . . . . . . . . . . . . . . . . . . . . .. 2.10 Controller structure selection results. N=100 . . . . . . . . . . . . . . . . . . . . . . . .. 2.11 Controller parameters dispersion from Monte-Carlo experiment. N=1000. . . . . . . . .. 2.12 Sample closed-loop step response. N=1000. . . . . . . . . . . . . . . . . . . . . . . . . .. 2.13 Dataset for process disturbance experiment. N=300. . . . . . . . . . . . . . . . . . . . .. 2.14 Closed-loop step responses. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 16 18 22 22 23 23 24 24 25 26 26 27 28 28 3.1 3.2 3.3 3.4 3.5 Assumed feedback control structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Model Matching block diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Lower noise bounds for different reference models and controller complexities (mmax ). .. Sparsity results for different ǫmax values. . . . . . . . . . . . . . . . . . . . . . . . . . . .. Results for bases selection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 31 33 39 40 42 4.1 4.2 4.3 4.4 4.5 4.6 4.7 Two-Degree-of-Freedom feedback control system . . . . . . . . . . . . . . . . . . . . . .. Block diagram for sensitivity tuning . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Block diagram for closed-loop model reference tuning . . . . . . . . . . . . . . . . . . . .. Bode plot results. 2 DoF controllers, example 1 . . . . . . . . . . . . . . . . . . . . . . .. Magnitude bode plots (loop specifications). 2 DoF controllers, example 2 . . . . . . . .. Results for sensitivity, example 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Results for reference model, example 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 43 46 47 51 52 52 53 5.1 Block diagram for Q tuning problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. s 5.2 Closed-loop with representation of the controller error Q̂−Q 1−M . . . . . . . . . . . . . . . .. 5.3 Reference models M1 class for diferrent α values . . . . . . . . . . . . . . . . . . . . . .. 5.4 Cost function for reference models M1 , evaluated for different α and mq parameters. . .. 5.5 Step response and bode plot of reference model M1 for α = 0.5 . . . . . . . . . . . . . .. 5.6 Results for 1000 controllers tuning via our approach.M1 . . . . . . . . . . . . . . . . . .. 5.7 Results for 1000 Q filters tuned via our approach. M1 . . . . . . . . . . . . . . . . . . .. 5.8 Step response and bode plot of reference model M2 . . . . . . . . . . . . . . . . . . . . .. 5.9 Cost function for reference model M2 for different mq values. . . . . . . . . . . . . . . .. 5.10 Results for 1000 controllers tuning via our approach.M2 . . . . . . . . . . . . . . . . . .. 57 60 61 62 62 63 63 64 65 65 8 5.11 Cascade control for the non-minimum phase plant . . . . . . . . . . . . . . . . . . . . .. 5.12 Results for 100 controllers tuning via our approach. Cascade control . . . . . . . . . . .. 5.13 2-DOF helicopter system. Courtesy : Quanser Inc. . . . . . . . . . . . . . . . . . . . . .. 5.14 2-DOF helicopter system connected with the stabilizing controller. . . . . . . . . . . . .. 5.15 Pitch and Yaw signals obtained for PRBS input . . . . . . . . . . . . . . . . . . . . . . .. 5.16 Pitch and Yaw positions via YK-DDC controller . . . . . . . . . . . . . . . . . . . . . .. 5.17 Cascade controller for the 2DOF helicopter . . . . . . . . . . . . . . . . . . . . . . . . .. 66 67 69 70 70 71 72 6.1 Steam distiller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.2 Model analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.3 Results in case I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.4 Results in case II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.5 Results in case III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.6 Effect of gamma and Pareto front . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.7 Effect on input trajectory produced by variations of Ktr and h parameters . . . . . . . .. 6.8 Optimal trajectory for non-automated plants . . . . . . . . . . . . . . . . . . . . . . . .. 6.9 Left: Ohmic heating system constructed. Right: Ohmic heating designed . . . . . . . .. 6.10 Ohmic heating camera with axial electric field . . . . . . . . . . . . . . . . . . . . . . . .. 6.11 Ohmic heating camera- Measures in [mm] . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.12 Condenser-clevenger designed. Linear measures in [mm] . . . . . . . . . . . . . . . . . .. 6.13 Experimental setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.14 Example for constant input power trajectory . . . . . . . . . . . . . . . . . . . . . . . .. 6.15 Essential oil accumulated in the Clevenger . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.16 Yield obtained in five experiments with constant input power trajectory . . . . . . . . .. 6.17 Analogous trajectories for steam distillation (a) and OAHD (b) . . . . . . . . . . . . . .. 6.18 Example for decreasing input power trajectory . . . . . . . . . . . . . . . . . . . . . . .. 6.19 Yield obtained in five experiments with decreasing input power trajectory . . . . . . . .. 6.20 Reaction curve in OAHD for u = 100% . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6.21 Reference model for temperature control in OAHD process . . . . . . . . . . . . . . . .. 6.22 Dataset used for both strategies in temperature control for OAHD process . . . . . . . .. 6.23 Tracking results for CbT controller in OAHD process . . . . . . . . . . . . . . . . . . . .. 6.24 Tracking results for YK-DDC controller in OAHD process . . . . . . . . . . . . . . . . .. 6.25 Tracking step results for CbT and YK-DDC controllers in OAHD process . . . . . . . .. 75 77 79 79 80 80 81 82 83 84 85 87 88 89 89 90 90 91 91 93 93 94 94 95 95 A.1 VRFT tuning problem block diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 9 Chapter 1 Introduction Currently, data acquisition technology permits to collect a large amount of measurements from industrial plants. When enough plant data are available in order to design a controller, there exist two main approaches in the scientific literature. In the standard model-based controller design procedure, a plant model is estimated (identified) and then employed to design a controller (two-step design). This controller is not necessarily optimal because the control loop performance is restricted by modeling errors. On the other hand, the available data can be employed to directly design a controller, avoiding the plant model estimation. This approach has been named Direct Data-Driven controllers (DDC) tuning. DDC is of particular interest in real-world practical applications. The interested reader can referred to Formentin et al. (2014), where an interesting comparison between Model-based controller design and DDC tuning is presented. There exist adaptive (iterative) and non-iterative DDC techniques, in the first ones the controller parameters are adjusted at each time instant, in the last ones the controller is designed based on the information contained in one batch of experimental data (one-shot), these approaches are known also as off-line techniques. In this work, we are interested in off-line techniques in order to avoid the typical problems of adaptive control, moreover, since a single set of data is employed in several model-based control design methods, a fair comparison between off-line DDC techniques and model-based design can be carried out. In the linear setting, some non-iterative controller design methods for direct data-driven controller tuning are shown and compared in (Van Heusden et al., 2011b), the studied methods are correlation approach (CbT), periodic errors in variables (EiV), the inverse controller (IC) and prediction error methods (PEM). All of them are constructed within a stochastic framework. A more recent review work of DDC methods can be found in (Hou and Wang, 2013). The authors perform a qualitative comparison and briefly explain each one technique. A different approach to solve the DDC problem follows a deterministic formulation using Set-membership techniques. There exist few results that follow this approach, one solution can be found in Cerone et al. (2017), also, for control of non-linear plants a technique named as Direct feedback control (DFK) has been developed in Novara et al. (2013a). DFK method can be used only in the restrictive case where the full state is measurable. The more relevant techniques for this thesis are virtual reference feedback tuning (VRFT) and correlation-based tuning (CbT). VRFT was proposed in (Guardabassi and Savaresi, 2000), it is a noniterative data-driven method that can be used to select the controller parameters for a LTI system, the problem is formulated as a controller parameters identification problem via introducing a virtual reference signal. Principal characteristics of VRFT are: (i) It is one-shot method in the linear case (noise free). (ii) The controller to find is linearly parametrized. (iii) When the measurements are noisy, the solution is addressed via instrumental variables. This requires a second experiment or a plant model. 10 (iv) The resulting optimization problem is convex. (v) There exist many extensions of VRFT, for instance, in (Campestrini et al., 2016) the MIMO case is tackled, and a non-linear framework is studied in (Formentin and Savaresi, 2011). In (Rojas, 2011) other extensions and applications of VRFT are studied, such author, indicates that in data-driven control it is difficult to find a controller that assures stability, and he proposes a stability test, this test requires previous identification of the plant, this is undesirable because in direct data-driven controller techniques the aim is to avoid developing a model of the plant. Regarding to CbT, it is worth to mention that exist two correlation based approaches for data-driven controller design, one of them is iterative (iCbT) (Karimi et al., 2002), another is non-iterative (CbT-GS) (van Heusden et al., 2011), as above mentioned, in this work our interest is limited to non-iterative techniques, that is why principal characteristics of CbT-GS are listed: (i) The problem is solved via model reference criterion, but it doesn’t need a virtual reference signal, (ii) In the same way that in VRFT, the controller is linearly parametrized, (iii) It is one-shot method and is better suited to deal with measurement noise (iv) In CbT-GS a sufficient conditions is proposed to guarantee closed-loop stability, (v) This method has not been extend to non linear framework. Few extensions on non-iterative CbT have been found in literature, one of them is about the tuning of controller parameters taking into account stability constraints for non-minimum phase plants (Matsuo et al., 2013), meanwhile in (Yubai et al., 2009) and (Usami, 2010) was addressed the MIMO case. In all the methods mentioned above, the main ingredients of a DDC problem are a set of input-output data generated by the plant to be controlled, a closed-loop reference model where performance specifications are embedded, and a given controller structure, usually parametrized by a fixed set of basis functions. When the set of bases is not consistent with the reference model, the resulting controller can yield to closed-loop instability. In this sense, in Kergus et al. (2019) a set of conditions to define the behaviours that the closed-loop can reach are proposed, in order to select an achievable reference model. According to Novara (2019) the drawbacks of DDC are: (i)Only sufficient conditions are currently available to guarantee closed-loop stability. (ii) Direct design is statistically less efficient than model-based design with correct model parametrization, and (iii) If the controller can be freely chosen, the problem of model parametrization is not skipped, but simply transformed into that of controller selection. Several applications of DDC have been reported in the literature, but none of them has considered the essential oil extraction process, this process is of high interest for this thesis. Essential Oil (EO) is an agro product which has a great prospect to be developed (Kusuma et al., 2018). The essential oils market accounted for USD 4.46 Billion in 2016, according to the Observatory of Economic Complexity (Simoes, 2018), and it is projected to reach USD 11.19 Billion by 2022. Essential oils are complex mixtures of volatile compounds extracted from a large number of plants. EO is stored in different parts of plants, for example in leaves, flowers, stems, roots, etc. There exist multiple essential oil extraction techniques (Stashenko, 2009). The most employed ones are hydro-distillation, steam distillation, and steam-water distillation. Some new techniques have been proposed, such as supercritical fluids extraction (SFE), microwave-assisted hydrodistillation (MWD), and ohmic-assisted hydrodistillation (OAHD). In this thesis, we are interested in OAHD since its main advantage over conventional distillation is that, shorter extraction times and lower energy consumption are obtained, thereby, it is environmentally friendly. The dissertation is organized in chapters and each one can be read independently. Figure 1.1 depicts the organization of the thesis, numbered by the respective chapter and their roles in the framework of this thesis. First, in Chapter 2, considering previous results by the author in Valderrama and Ruiz (2014) a setmembership approach to give solution to DDC design problem is proposed, our formulation is named Set-Membership tuning (SMT). In SMT, the Unknown But Bounded (UBB) noise structure affecting the 11 Applications & benckmarks Set-Membership approach (SISO plants). Active suspension Chapter 2. Youla-Kucera parametrization (SISO plants). Chapter 5. Set-Membership (Limited- complexity controllers) Flexible transmission Chapter 3. Set-Membership (Extension to the MIMO case) Chapter 5. 2DoF Helicopter Set-Membership (2 DoF controller structuctures) Chapter 4. Essential oil extraction processes Controlling the heating curve in OAHD Chapter 6. An optimal control approach steam distillation Experimental tests for input power trajectories in OAHD Chapter 6. Chapter 6. OAHD design and construction Chapter 6. Figure 1.1: Thesis organization experimental data has been assumed, and a proper data-based criterion is stated according to the reference model criterion problem. Such formulation allows to state hard bounds on controller parameters via linear programming techniques. We compare our approach with the Virtual Reference Feedback Tuning (VRFT) technique which assumes an stochastic description of the unknown signals. The two approaches are evaluated on an experimental case study, consisting of the controller tuning for an active suspension system. The results are obtained via a Monte-Carlo experiment, one hundred controllers are tuned by both methods employing different noise realizations and the performance is measured experimentally. Results show that both approaches offer a similar performance when the size of the dataset is much larger than the dimension of the controller parameters vector. However, for reduced datasets, the SMT approach gives consistent results while the VRFT method is not able to extract useful information. The same behavior is observed when the two approaches are applied to datasets affected by process disturbances. It is observed that the Root Mean Squared Error of the resulting loops can be up to 30 times 12 lower using the Set Membership method for reduced datasets. The results of this Chapter were published in (Valderrama and Ruiz, 2019).	40,708
https://openalex.org/W4205531402	OpenAlex	Open Science	CC-By	2,021	DISTORÇÕES CAUSADAS PELA FALTA DE CALIBRAÇÃO RADIOMÉTRICA E EFEITOS ATMOSFÉRICOS NA ANÁLISE DE DADOS DE SENSORIAMENTO REMOTO ORBITAL	André WELTER	Portuguese	Spoken	8,892	17,413	Resumo O artigo investiga distorções nas informações extraídas de dados de sensores remotos quando não são feitas calibração radiométrica e correção de efeitos atmosféricos. Foram comparados valores de pixels em unidades de Níveis Digitais, reflectância sem correção (reflectância no topo da atmosfera) e reflectância com correção (reflectância de superfície) de efeitos atmosféricos, obtidos de alvos específicos (área queimada, corpo d’água, cultura agrícola, vegetação nativa e solo exposto) de uma imagem multiespectral adquirida pelo satélite Landsat 7, órbita 226, ponto 071, sobre a região de Cuiabá – MT, do dia 28/09/2002. Em seguida, foram comparados índices de vegetação da diferença normalizada obtidos a partir dos dados não calibrados (ND) e reflectâncias de superfície (com correção) a fim de verificar o impacto da utilização dos dados brutos. A análise demonstra a importância de se considerar os efeitos atmosféricos nos estudos realizados com informações obtidas por sensores remotos orbitais (que, em muitos casos, não são corrigidas), uma vez que as assinaturas espectrais extraídas dos dados sem tratamento destoam significativamente do esperado, segundo a literatura especializada. Palavras-chave: Detecção remota. Plataformas orbitais. Espalhamento atmosférico. Reflectância de superfície. Reflectância no topo da atmosfera. DISTORÇÕES CAUSADAS PELA FALTA DE CALIBRAÇÃO RADIOMÉTRICA E EFEITOS ATMOSFÉRICOS NA ANÁLISE DE DADOS DE SENSORIAMENTO REMOTO ORBITAL André WELTER1 Thiago STATELLA2 Geneci Marques de LIMA3 Geneci Marques de LIMA3 1 Serviço de Cartografia, Instituto Nacional de Colonização Agrária – INCRA. E-mail: andrelwgeo@gmail.com 2 Departamento de Área de Construção Civil, Instituto Federal de Educação, Ciência e Tecnologia de MT – IFMT. E-mail: thiago.statella@cba.ifmt.edu.br 3 Departamento de Área de Construção Civil, Instituto Federal de Educação, Ciência e Tecnologia de MT – IFMT. E-mail: limageneci46@gmail.com 3 Departamento de Área de Construção Civil, Instituto Federal de Educação, Ciência Tecnologia de MT – IFMT. E-mail: limageneci46@gmail.com 1 Serviço de Cartografia, Instituto Nacional de Colonização Agrária – INCRA. E-mail: andrelwgeo@gmail.com 2 2 Departamento de Área de Construção Civil, Instituto Federal de Educação, Ciência Tecnologia de MT – IFMT. E-mail: thiago.statella@cba.ifmt.edu.br 3 1 Serviço de Cartografia, Instituto Nacional de Colonização Agrária – INCRA. E-mail: andrelwgeo@gmail.com 2 Departamento de Área de Construção Civil, Instituto Federal de Educação, Ciência e Tecnologia de MT – IFMT. E-mail: thiago.statella@cba.ifmt.edu.br Abstract This paper investigates the distortions in information extracted from remote sensing data when radiometric calibration and atmospheric effects correction are not carried out. For the comparison, we used pixel values from raw data in Digital Numbers, from the top of the atmosphere reflectance (TOA) uncorrected G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 1 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de data, and from the surface reflectance data (SR), gathered from selected targets (such as burned crop, native vegetation, rivers and bare soil) in a multispectral image acquired by the Landsat 7 satellite, row 226, path 071, depicting the Cuiabá city in MT state, on 28/09/2002. Next, we compared Normalized Difference Vegetation Index calculated from the uncalibrated data (DN) and from the surface reflectances (atmosphere corrected) so that we could verify the errors when using raw data. The analysis shows the importance of the atmospheric correction prior the analysis when dealing with orbital sensors (in several cases not corrected), given that the spectral signatures of the targets were severely drifted from the ones expected according to the specialized literature. Keywords: Remote detection. Orbital platform. Atmospheric reflection. Surface reflectances. Top of the atmosphere reflectance. Resumen El artículo investiga las distorsiones en la información extraída de los datos del sensor remoto cuando no se realizan la calibración radiométrica y la corrección de los efectos atmosféricos. Los valores de píxeles se compararon en unidades de niveles digitales, reflectancia sin corrección (reflectancia en la parte superior de la atmósfera) y reflectancia con corrección (reflectancia superficial) de los efectos atmosféricos, obtenidos de objetos específicos (quemadas, cuerpo de agua, cultivo, vegetación nativa y suelo expuesto) de una imagen multiespectral adquirida por el satélite Landsat 7, órbita 226, punto 071, sobre la región de Cuiabá - MT, del 28/09/2002. Además, se compararon los índices de vegetación de diferencia normalizada obtenidos de los datos no calibrados (ND) y las reflectancias de la superficie (con corrección) para verificar el impacto del uso de datos sin procesar. El análisis demuestra la importancia de considerar los efectos atmosféricos en estudios realizados con información obtenida por sensores orbitales remotos (que, em muchos casos, no se corrigen), ya que las caracteristicas espectrales extraídas de los datos sin tratamiento difieren significativamente de lo esperado, según la literatura especializada. g p g p Palabras clave: Detección remota. Plataformas orbitales. Dispersión atmosférica. Reflectancia superficial. Reflectancia en la parte superior de la atmósfera. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . INTRODUÇÃO Atualmente, os métodos de Sensoriamento Remoto tornaram-se uma ferramenta indispensável para diversas atividades práticas que estão inseridas no cotidiano das pessoas que vivem no meio urbano e rural (como monitoramento de culturas agrícolas, análise para regularização fundiária, G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 2 Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital mapas de localização e traçado de rotas, etc.). Dados de sensores remotos são utilizados para otimizar custos de aquisição de informações espectrais sobre grandes áreas, facilitar planejamento, monitoramento ambiental ou de produção, visando obter melhores resultados e maior controle sobre os processos. Contudo, quando não tratadas adequadamente, imagens de satélite fatalmente levarão o usuário a equívocos e análises incorretas. As imagens de satélite resultam da leitura obtida, por sensores orbitais, da Radiação Eletromagnética (REM) refletida pela superfície terrestre (radiância). Esta energia, quando na região do espectro visível, sofre espalhamento e absorção pelos vários constituintes atmosféricos antes de chegar ao sensor. Os menores comprimentos de onda na região do visível sofrem espalhamento molecular (ou de Rayleigh), causado por moléculas de gás. Partículas de poeira e fumaça atenuam os maiores comprimentos de onda do espectro visível, num efeito conhecido como espalhamento Mie. Ainda, há o espalhamento não seletivo, causado por vapor d’água, e cuja magnitude independe do comprimento de onda da energia eletromagnética. Apesar disso, alguns trabalhos não apresentam a correção atmosférica em seus dados (ANDRADE et al., 2018; SILVIA et al., 2011), alguns aplicam métodos aproximados (LEITE et al., 2017), como o Dark Object Subtraction (que opera no contradomínio dos ND) e, ainda, há trabalhos em que a calibração radiométrica e correções atmosféricas não são aplicadas (RIBEIRO et al., 2018; ZANZARINI et al., 2013; VELASCO et al., 2007). Não raro, o contradomínio das imagens utilizadas é um intervalo de Níveis Digitais (ND, ou tons de cinza), quando deveria ser um intervalo de radiâncias (em ) ou reflectâncias, que são grandezas com significado físico, diretamente correlacionadas às características físico-químicas dos alvos. Este artigo tem o objetivo de demonstrar as distorções ocorridas nas análises de imagens quando a calibração radiométrica e efeitos atmosféricos são negligenciados. WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de de uma imagem multiespectral adquirida pelo satélite Landsat 7 (órbita/ponto 226/071), sobre Cuiabá – MT e região, no dia 28/09/2002, período do ano de estiagem, sem a presença de nuvens na cena. Além disso, foram comparados índices de vegetação por diferença normalizada (Normalized Difference Vegetation Index - NDVI) obtidos a partir das reflectâncias no topo da atmosfera e de superfície a fim de verificar o impacto da utilização dos dados não corrigidos dos efeitos da atmosfera em vez dos dados tratados. Uma vez que toda cena adquirida por plataforma orbital possuirá distorções causadas pela atmosfera (a depender apenas do tipo de mecanismo espalhador em ação), tais efeitos são bem caracterizados neste caso de estudo, não havendo a necessidade de se processar um conjunto maior de imagens. O trabalho está organizado da seguinte forma: no item 2 são abordados aspectos teóricos sobre a correção atmosférica de dados de sensores remotos, no item 3 apresenta-se a área de estudo, no item 4 são dados detalhes do método empregado para analisar as imagens, no item 5 são exibidos os resultados e, por fim, no item 6 faz-se a conclusão do trabalho, que é seguida do item de referências bibliográficas. INTRODUÇÃO Isto é demonstrado graficamente e quantitativamente pela comparação entre valores de pixels em unidades de ND, reflectância no topo da atmosfera (TOA – Top of the Atmosphere) e reflectância de superfície (RS, ou seja, reflectância corrigida de efeitos atmosféricos), obtidos de alvos específicos (área queimada, corpo d’água, cultura agrícola, vegetação nativa e solo exposto), G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 3 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de CORREÇÃO DE EFEITOS ATMOSFÉRICOS Segundo Novo (2010), no Sensoriamento Remoto da superfície terrestre a principal fonte de radiação eletromagnética é o Sol. A radiação gerada pelas reações em seu interior se propaga na forma de ondas ou partículas em todas as direções e está sujeita à interação com a atmosfera e seus componentes. Os componentes da atmosfera que mais influenciam na absorção da radiação eletromagnética que efetivamente chega à superfície da Terra são: o vapor d’água (H2O), o Oxigênio (O2), o Ozônio (O3) e o Gás Carbônico (CO2), que formam uma atmosfera opaca que barra parte radiação antes de ocorrer a interação com os objetos da superfície terrestre. Para Antunes et al. (2003) a reflectância obtida da mistura de sinal da atmosfera e da superfície é dita reflectância aparente ou reflectância no topo da atmosfera (TOA), sendo a reflectância definida como a razão entre o fluxo refletido pelo alvo e o fluxo incidente no mesmo (NOVO, 2010). G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 4 Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital A atmosfera absorve ou espalha a radiação eletromagnética de forma diferenciada em função dos seus comprimentos de onda. As partículas presentes na atmosfera, como aerossóis, poeiras, moléculas de diversos gases com diferentes tamanhos interferem na radiação que chega aos alvos terrestres ou ao sensor. Ou seja, a energia eletromagnética ao atingir a atmosfera é por esta espalhada, e parte desta energia espalhada retorna para o espaço, vindo a contaminar a energia refletida ou emitida pela superfície e que é detectada pelos sensores orbitais (PONZONI et al., 2015). A diminuição do contraste entre objetos de uma mesma cena, a redução da possibilidade de identificação de pequenos objetos ou a diferenciação entre objetos de uma cena são consequência dos efeitos da atmosfera nas imagens obtidas por sensoriamento remoto, além da clara alteração na radiância medida pelo sensor. Novo (2010) entende que a correção dos efeitos atmosféricos é importante especialmente em três casos específicos: quando se tem a necessidade de recuperar o valor da grandeza radiométrica medida, ou seja, quando se quer conhecer a reflectância, emitância ou retroespalhamento do objeto em estudo, para poder utilizar estes valores em modelos empíricos ou teóricos; quando se pretende utilizar algoritmos que se baseiam em operações aritméticas entre bandas, como o NDVI; ou quando se objetiva comparar imagens de diferentes datas em termos de propriedades dos objetos na cena. A correção atmosférica é essencial para as análises de imagens de satélite, devido ao fato que a interação entre os gases e as partículas presentes na atmosfera com o fluxo de energia eletromagnética vinda do sol prejudica a passagem das ondas eletromagnéticas através da atmosfera, causando interferências. Essas interferências acontecem devido a três tipos de interações atmosféricas na energia reemitida por um alvo, quando este é visualizado do espaço. Estas alterações são conhecidas como espalhamento, absorção e reflectância (NOVO, 2010). Existem três formas de espalhamentos: Rayleigh, Mie e não-seletivo. O espalhamento Rayleigh é produzido pelas partículas de gases constituintes da atmosfera cujos diâmetros são menores que o λ (comprimento de onda) da radiação. O espalhamento Mie, provocado por fumaça e poeira, ocorre quando o G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital 3 6 6 Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital 3 6 5 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de diâmetro das partículas presentes na atmosfera é aproximadamente igual ao λ da radiação. E o espalhamento não-seletivo, provocado por vapor d’água, ocorre quando o diâmetro das partículas espalhadoras é muito maior que os comprimentos de onda, neste caso a REM de diferentes comprimentos de onda será espalhada com igual intensidade. Todo espalhamento ocorre pela absorção e remissão aleatória de radiação por átomos ou moléculas presentes na atmosfera (JENSEN, 2009). Vários métodos para correção destes efeitos são encontrados na literatura, desde os mais simples, de correção relativa, como o DOS (Dark Object Subtraction), cujo contradomínio são ND (e partem da premissa de que em toda cena existe um alvo com radiância nula, o que não se verifica em muitas situações), até os mais precisos, que utilizam algoritmos para modelagem da transferência radiativa da atmosfera, e que operam sobre radiância e/ou reflectância (CHAVEZ, 1988; MATTHEW et al., 2002; RUMORA et al., 2020). Estes últimos são conhecidos como de correção absoluta, e requererem a prévia calibração radiométrica dos dados brutos, processo pelo qual os valores de ND (quantização das radiâncias) são convertidos em radiância (valor com significado físico realmente medido pelo sensor). Isso pode ser feito a partir dos parâmetros de calibração radiométrica dos sensores, normalmente disponíveis para usuários no caso de sensores orbitais por meio de relatórios técnicos e/ou documentação fornecida pelo fabricante das câmeras. Para as imagens do sensor ETM+ a bordo do Landsat 7, utilizado nesta pesquisa, é comum a adoção do modelo 6S (Second Simulation of the Satellite Signal in the Solar Spectrum), baseado no cálculo da transferência radiativa na atmosfera. Segundo Antunes et al. (2003) o modelo 6S é uma versão aprimorada do 5S (Simulação do sinal de satélite no espectro solar), feito pelo Laboratoire d'Optique Atmospherique. O modelo 6S foi desenvolvido por Vermote et al. (1997), para simular a radiância que chega nos sensores em bandas dentro do espectro solar, entre 0,25 m a 4,00 m. Sendo assim, pode-se calcular a reflectância de superfície (RS) do alvo, isto é, a reflectância livre dos efeitos de espalhamento atmosférico. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Na figura 1, estão marcados, com anotações em vermelho, os alvos estudados: área queimada, corpo d’água, cultura agrícola, vegetação nativa e solo exposto. A imagem é exibida na composição B(3)G(4)R(5), para realçar a resposta da vegetação no infravermelho próximo e também áreas queimadas no infravermelho de ondas curtas. Esta cena, nesta data, foi escolhida por apresentar uma grande diversidade de alvos com características espectrais distintas, tais como áreas de área queimada, corpos d’água, cultura agrícola, vegetação nativa (de grande porte) e solo exposto, bem como uma quantidade muito pequena de nuvens (concentradas no canto superior direito da cena). CENA DE ESTUDO A imagem multiespectral escolhida para o trabalho foi a adquirida pelo sensor ETM+ a bordo do satélite Landsat 7, órbita/ponto 226/071, na data de 28/09/2002, na região de Cuiabá – MT, nas bandas 1 (azul, 0,45 – 0,52 m), 2 (verde, 0,52 – 0,60 m), 3 (vermelho, 0,63 – 0,69 m), 4 (infravermelho próximo, 0,76 – 0,90 m), 5 (infravermelho médio, 1,55 – 1,75 m) e 7 (infravermelho médio, 2,08 – 2,35 m). Cada canal possui resolução espacial de 30 m e resolução radiométrica de 8 bits (ou 256 níveis de quantização). A resolução temporal da plataforma é de 16 dias. A imagem foi escolhida por apresentar diversos tipos diferentes de cobertura do solo sem cobertura de nuvens, além de estar disponível no servidor do USGS nas versões sem calibração radiométrica e com reflectância de superfície. A figura 1 exibe a imagem da área de estudo, no sistema de referência WGS84, e sistema de projeção UTM, fuso 21 S. Figura 1- Imagem Landsat 7 (órbita/ponto 226/071) da área de estudo, composição B(3)G(4)R(5), sistema de referência WGS84, sistema de projeção UTM, fuso 21 Figura 1- Imagem Landsat 7 (órbita/ponto 226/071) da área de estudo, composição B(3)G(4)R(5), sistema de referência WGS84, sistema de projeção UTM, fuso 21 Fonte: Autoria própria Figura 1- Imagem Landsat 7 (órbita/ponto 226/071) da área de estudo, composição B(3)G(4)R(5), sistema de referência WGS84, sistema de projeção UTM, fuso 21 3)G(4)R(5), sistema de referência WGS84, sistem UTM, fuso 21 Fonte: Autoria própria Fonte: Autoria própria G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 7 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . MÉTODO A figura 2 exibe um fluxograma com cada uma das etapas executadas no presente trabalho. Figura 2 - Fluxograma das etapas executadas Figura 2 - Fluxograma das etapas executadas Fonte: Autoria própria Fonte: Autoria própria Fonte: Autoria própria Foram adquiridas duas versões da mesma cena Landsat 7 (sensor ETM+) junto ao Serviço Geológico dos Estados Unidos (USGS, disponível em https://earthexplorer.usgs.gov). Uma delas em unidades de ND (sem calibração radiométrica e sem correção atmosférica) e a outra com correção atmosférica (em unidades de reflectância de superfície, portanto). As imagens Landsat 7 com correção atmosférica são processadas (USGS, 2019a; 2019b) com o G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 8 . Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital LEDAPS (Landsat Ecosystem Disturbance Adaptive Processing System). No processo, utiliza-se o código 6S (Second Simulation of the Satellite Signal in the Solar Spectrum) como modelo de transferência radiativa da atmosfera para a conversão dos dados brutos em reflectância de superfície. LEDAPS (Landsat Ecosystem Disturbance Adaptive Processing System). No processo, utiliza-se o código 6S (Second Simulation of the Satellite Signal in the Solar Spectrum) como modelo de transferência radiativa da atmosfera para a conversão dos dados brutos em reflectância de superfície. A versão bruta da imagem, em unidades de ND (com resolução radiométrica de 8 bits, isto é, com tonalidades de cinza variando dentro do intervalo [0, 255]), foi submetida a um processamento para converter seu contradomínio em unidades de reflectância no topo da atmosfera, no intervalo [0,1]. Previamente, aplicou-se uma filtragem por mediana (máscara 3x3), para eliminar diversos ruídos (erros de bit, inerentes ao próprio sistema sensor ou causados em decorrência de falhas no processo de transmissão dos dados) presentes na cena. Em seguida, aplicaram-se os parâmetros de calibração radiométrica (CHANDER et al., 2009; GÜRTLER et al. 2005) para a conversão de ND em radiância espectral, em unidades de , empregando-se a equação 1 (USGS, 2019b): ( ) (1) ( ) (1) ( ) ( ) (1) (1) Em que é a radiância espectral ( ) na banda , e são, respectivamente, as radiâncias máxima e mínima esperadas para a banda e ND são os valores da imagem de entrada. Os valores de radiância, então, foram convertidos em reflectância no topo da atmosfera (TOA) pela seguinte equação (USGS, 2019b; PONZONI et al., 2015): . (2) . (2) Em que é a reflectância espectral na banda  no topo da atmosfera, d é a distância Terra-Sol média, em unidades astronômicas (UA), é a irradiância solar média (PONZONI et al., 2015; NOVO, 2010) no topo da G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 6 9 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Em que é a reflectância no infravermelho próximo e é a reflectância no vermelho. Como se percebe pela equação 4, o NDVI é calculado a partir das reflectâncias nas bandas do vermelho e infravermelho próximo. A razão de bandas utilizando dados brutos de ND não é, em rigor, um índice de vegetação. Isso porque, ao não se fazer a calibração radiométrica dos dados, negligencia-se a função de transferência dos CCD, que é particular para cada uma das bandas (ganhos e offsets diferentes). Entretanto, como vários autores assim o fazem, tal razão de bandas será também chamada neste trabalho de NDVI, apenas por simplicidade. Os efeitos desse erro (isto é, a não adoção de reflectâncias na equação 4) serão analisados no próximo item de resultados, quando serão comparados os índices formados a partir de RS e ND. RESULTADOS E DISCUSSÃO A seguir, no item 5.1, são apresentadas e discutidas as assinaturas espectrais obtidas para cada alvo de interesse ao longo da resolução espectral do Landsat 7, calculadas para valores de ND, TOA e RS. No item 5.2, apresentam- se os NDVI calculados para valores de ND e RS. WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de atmosfera (em ) para a banda  e  é o ângulo zenital solar no centro da cena no momento da aquisição. atmosfera (em ) para a banda  e  é o ângulo zenital solar no centro da cena no momento da aquisição. A distância Terra-Sol média d pode ser obtida da seguinte relação (STEIN et al., 2002): * ( ) + (3) (3) Em que DDA é o dia do ano, contado a partir do dia 01 de janeiro do ano da tomada da cena. Em que DDA é o dia do ano, contado a partir do dia 01 de janeiro do ano da tomada da cena. A partir de então, estavam disponíveis três versões da mesma cena: os dados brutos, em unidades de ND, uma imagem de reflectância no topo da atmosfera (TOA, sem correção atmosférica) e outra imagem de reflectância de superfície (RS, corrigida de efeitos atmosféricos pela aplicação do algoritmo 6S). Para comparação destes três dados, foram selecionadas as seguintes classes de informação: área queimada, corpo d’água, cultura agrícola, vegetação nativa e solo exposto, identificadas a partir de interpretação visual de composições coloridas das cenas. Amostras com área de 9 pixels (3 x 3) de cada uma das classes foram coletadas, e seus valores médios foram calculados nas bandas 1, 2, 3, 4, 5 e 7. Optou-se por distribuições pequenas (9 pixels) para se evitar que a variância das amostras dentro de cada retângulo fosse grande. Além disso, a média garante que erros de registro entre as bandas não interfiram nas análises posteriores (se apenas o valor de um pixel fosse coletado, não seria possível assegurar que ele representasse exatamente a mesma área de 30 x 30 m no terreno em todas as bandas). Em seguida, foram calculados índices de vegetação NDVI para os dados de reflectância de superfície e para os dados em níveis digitais, conforme a equação 4 (JENSEN, 2009): (4) G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 0 Comparação entre curvas espectrais As curvas espectrais dos alvos área queimada, corpo d’água, cultura agrícola, vegetação nativa e solo exposto (marcados na figura 1) foram obtidas a partir da interpretação das cenas e extração de valores de pixels (médias de uma região de 3 x 3 pixels centrada em cada classe de informação) em todas as 7 bandas do sensor. Para cada alvo, foram construídas três curvas com base em seus valores nas imagens bruta (ND), de reflectância no topo da atmosfera (TOA) e reflectância de superfície (RS). Para facilitar a visualização dos valores, as quantidades de ND foram normalizadas para o intervalo [0,1], de maneira que pudessem ser exibidos em mesma escala que os dados de reflectância nos gráficos. A figura 3 mostra os valores para o alvo queimada. Este alvo tem como característica espectral baixos valores nas bandas do visível (bandas 1-3) e G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 1 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de infravermelho próximo (banda 4). Da análise desta figura percebe-se que a curva de ND é completamente diferente as curvas de reflectância, e não condiz com o comportamento esperado. Ao analisar as curvas de reflectância para queimada, é possível perceber o efeito da atmosfera na curva de TOA, fazendo com que as bandas apresentem valores mais elevados que aqueles da RS, especialmente na banda do azul, em que o espalhamento atmosférico é maior. Figura 3 - Gráfico dos valores de ND, TOA e RS por banda do alvo queimada. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 INTENSIDADE Figura 3 - Gráfico dos valores de ND, TOA e RS por banda do alvo queimada. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 INTENSIDADE Figura 3 - Gráfico dos valores de ND, TOA e RS por banda do alvo queimada Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Figura 3 - Gráfico dos valores de ND, TOA e RS por banda do alvo queimada. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 , REFLECTÂNCIA DE SUPERFÍCIE Fonte: Autoria própria A figura 4 exibe os valores obtidos para o alvo corpo d’água sem sedimentos suspensos. 3 7 2 G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Figura 4 - Gráfico dos valores de ND, TOA e RS por banda do alvo corpo d’água. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Figura 4 - Gráfico dos valores de ND, TOA e RS por banda do alvo corpo d’água. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE Figura 4 - Gráfico dos valores de ND, TOA e RS por banda do alvo corpo d’água. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 7 Banda 5 O comportamento que mais destoa entre as três curvas na figura 4 está em ND, principalmente nas bandas 1, 2 e 3 (bandas do visível), com comportamento parecido a partir da banda 4 até a banda 7 (bandas do infravermelho próximo e médio). O comportamento esperado deste tipo de alvo pode ser visto na figura 5, a qual exibe o comportamento espectral de alguns tipos de alvos. Figura 5 - Comportamento espectral esperado dos alvos Grama, Concreto, Sol Arenoso, Telhados, Solo Exposto, Asfalto, Grama Artificial e Água Limpa. Fonte JENSEN (2009) Figura 5 - Comportamento espectral esperado dos alvos Grama, Concreto, Solo Arenoso, Telhados, Solo Exposto, Asfalto, Grama Artificial e Água Limpa. Fonte: JENSEN (2009) G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 Fonte: Modificado de Jensen (2009) Fonte: Modificado de Jensen (2009) Fonte: Modificado de Jensen (2009) Fonte: Modificado de Jensen (2009) G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital 2 0 2 0 . G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 3 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Na figura 5, percebe-se que a reflectância da água é muito baixa, se comparada com outros alvos, e, por isso, ela tende a se apresentar bastante escura nas imagens de satélite. Sua reflectância é nula nas bandas do infravermelho, devido à absorção da radiação eletromagnética nestes comprimentos de onda. A água sem sedimentos suspensos apresenta maior reflectância na região espectral do azul, por causa do espalhamento molecular. Na figura 4, ao se comparar as curvas de TOA e RS percebe-se que o espalhamento atmosférico provoca uma reflectância maior em todas as bandas, mas mais acentuadamente nos menores comprimentos de onda. Além disso, a forma da curva composta por valores de ND é completamente descaracterizada por causa da falta de calibração radiométrica dos dados e correção atmosférica. Nas figuras 6 e 7 exibem-se as curvas obtidas para os alvos vegetação nativa (de grande porte) e cultura agrícola. As características das curvas de TOA e RS são parecidas no infravermelho, destoando mais no visível. Contudo, é evidente a interferência atmosférica nos dados de reflectância TOA, assim como, do ganho do sensor para o caso do ND. O aspecto da curva obtida com os dados de ND é bastante diferente das curvas de SR e TOA. Nos valores absolutos das curvas de SR e TOA, nas bandas 1, 2 e 3 (visível), observa-se que só na reflectância de superfície (RS) o comportamento está de acordo com o esperado para vegetação (conforme figura 5), ou seja, valores baixos nas bandas do azul e vermelho, e maiores na banda do verde. Isso deve ocorrer para a vegetação sadia, que absorve energia nas faixas do azul e vermelho para fotossíntese, e reflete mais no verde. Alterações neste comportamento são indícios de estresse da planta (causado, por exemplo, pela falta de irrigação, de nutrientes, por pragas, etc.). Percebe-se que efeitos atmosféricos alteram a assinatura esperada para a vegetação, especialmente na banda do azul. Uma análise desta curva levaria a conclusões erradas sobre o estado da vegetação, portanto. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 4 Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Figura 6 - Gráfico dos valores de ND, TOA e RS por banda do alvo cultura agrícola. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria d l l fi 8 i d Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Figura 6 - Gráfico dos valores de ND, TOA e RS por banda do alvo cultura agrícola. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE Figura 6 - Gráfico dos valores de ND, TOA e RS por banda do alvo cultura agrícola. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Figura 6 - Gráfico dos valores de ND, TOA e RS por banda do alvo cultura agrícola. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Figura 6 - Gráfico dos valores de ND, TOA e RS por banda do alvo cultura agrícola. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 Fonte: Autoria própria REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Figura 8 - Gráfico dos valores de ND, TOA e RS por banda do alvo solo exposto. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 0,60 0,70 0,80 INTENSIDADE Figura 8 - Gráfico dos valores de ND, TOA e RS por banda do alvo solo exposto. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 0,60 0,70 0,80 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA Figura 8 - Gráfico dos valores de ND, TOA e RS por banda do alvo solo exposto. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Figura 8 - Gráfico dos valores de ND, TOA e RS por banda do alvo solo exposto. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,10 0,20 0,30 0,40 0,50 0,60 0,70 0,80 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE Fonte: Autoria própria Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Fonte: Autoria própria Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE REFLECTÂNCIA DE SUPERFÍCIE REFLECTÂNCIA NO TOPO ATMOSFERA NÚMERO DIGITAL Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. Os valores de ND foram normalizados para o intervalo [0, 1] para melhor visualização Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. Os valores de ND foram normalizados para o intervalo [0, 1] para melho visualização Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. Os valores de ND foram normalizados para o intervalo [0, 1] para melho visualização Figura 7 - Gráfico dos valores de ND, TOA e RS por banda do alvo vegetação nativa. Os valores de ND foram normalizados para o intervalo [0, 1] para melh visualização Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35 0,40 INTENSIDADE Banda 1 Banda 2 Banda 3 Banda 4 Banda 5 Banda 7 REFLECTÂNCIA NO TOPO ATMOSFERA No caso do alvo solo exposto, na figura 8, mais uma vez, a curva de ND difere completamente do comportamento esperado, caracterizado pelas curvas de reflectância (conforme figura 5). A presença de efeitos atmosféricos fica evidente na comparação das curvas de TOA e RS. Na curva de TOA os valores de reflectância são maiores em todas as bandas. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 5 Índice de Vegetação por Diferença Normalizada – NDVI A fim de analisar qualitativamente e quantitativamente o impacto da falta de calibração radiométrica e correção atmosférica nas informações extraídas de imagens de sensoriamento remoto, foram calculados NDVI a partir da imagem de ND e da imagem de RS. Em consonância com as comparações feitas nas figuras (3 a 8), em que todos os alvos sem correção se comportaram de maneira diferente em relação às assinaturas espectrais esperadas (nos dados corrigidos), nota-se grande discrepância entre os NDVI. Os mapas temáticos gerados a partir dos índices são exibidos na figura 9. A paleta de cores e as classes dos mapas foram configuradas para ser coincidentes. 3 7 6 G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Figura 9 – Mapas temáticos gerados a partir dos NDVI calculados com valores de ND (A) e RS (B). As classes temáticas foram definidas de maneira idêntica para ambos, conforme a legenda Figura 9 – Mapas temáticos gerados a partir dos NDVI calculados com valores de ND (A) e RS (B). As classes temáticas foram definidas de maneira idêntica para ambos, conforme a legenda Figura 9 – Mapas temáticos gerados a partir dos NDVI calculados com valores de ND (A) e RS (B). As classes temáticas foram definidas de maneira idêntica para ambos, conforme a legenda Fonte: Autoria própria Fonte: Autoria própria Os valores de um NDVI ocupam o intervalo [-1,1]. Nestes índices, pixels de vegetação sadia apresentam valores próximos de 1. Solo exposto, geralmente, apresenta valores positivos baixos, entre 0,3 e 0. Valores negativos são produzidos por sombra e água. O intervalo de valores predominantes no NDVI da imagem de ND (figura 9A) é de [-0,4, -0,2], que corresponderia a pixels de sombra ou água, enquanto na imagem com correção atmosférica (RS) o intervalo predominante (figura 9B) é de [0,2, 0,4], correspondendo a solo exposto. Existe, portanto, significativa diferença para os resultados de NDVI. Esta diferença é causada pela interferência incontestável da atmosfera na reflectância em produtos gerados por sensoriamento remoto orbital, além da falta de calibração radiométrica. Índice de Vegetação por Diferença Normalizada – NDVI A seguir, nas figuras 10 até 14 são exibidos os alvos analisados em composição colorida B(3)G(4)R(5), e os NDVI calculados a partir dos dados de ND e RS. Estes estão representados em mapas temáticos, para os quais uma mesma legenda de classes foi empregada. De maneira geral, os NDVI são G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 7 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de bastante diferentes para os dados brutos e corrigidos, provando que análises feitas sobre razões de bandas calculadas a partir de dados brutos (erroneamente chamados de NDVI) conduzem a conclusões erradas sobre o comportamento dos alvos estudados. Figura 10 – Classe de informação: queimada. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Figura 11 – Classe de informação: corpo d’água. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Figura 10 – Classe de informação: queimada. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) reflectância de superfície em (C) Fonte: Autoria própria Figura 11 – Classe de informação: corpo d’água. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir reflectância de superfície em (C) Figura 11 – Classe de informação: corpo d’água. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Fonte: Autoria própria 3 7 8 G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Figura 14 – Classe de informação: solo exposto. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Figura 14 – Classe de informação: solo exposto. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir reflectância de superfície em (C) Da análise das figuras 10 a 14 fica evidente que a prática comum de se utilizarem os dados de ND diretamente na equação 4 produz resultados enviesados. As classes de informação são completamente descaracterizadas nos NDVI calculados dos valores de ND. Informações extraídas de tais produtos estarão comprometidas e a análise destas informações levará a conclusões erradas sobre o comportamento dos alvos apresentados. Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Figura 12 – Classe de informação: cultura agrícola. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Figura 13 – Classe de informação: vegetação nativa. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Figura 12 – Classe de informação: cultura agrícola. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Fonte: Autoria própria Figura 13 – Classe de informação: vegetação nativa. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir d reflectância de superfície em (C) Figura 13 – Classe de informação: vegetação nativa. Composição colorida B(3)G(4)R(5) em (A), NDVI a partir dos dados brutos em (B) e NDVI a partir da reflectância de superfície em (C) Fonte: Autoria própria Fonte: Autoria própria G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 7 9 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Somados à falta de calibração radiométrica, os efeitos atmosféricos alteram as curvas de reflectância, como demonstrado. Trabalhos que utilizem os valores de reflectância em suas análises podem ser comprometidos e não representar fidedignamente a realidade em campo, caso não levem em consideração as interferências causadas pela atmosfera, como absorção e espalhamento. Foi demonstrado que, apenas quando foram utilizados dados de reflectância de superfície, obtidas pela utilização do algoritmo 6S, as curvas de reflectância espectral dos alvos foram coincidentes com as esperadas. Por fim, na geração de mapas temáticos, a partir do cálculo de NDVI dos dados de ND e RS, observa-se grande diferença nos intervalos de classificação, que geram um aspecto visual diferente para os índices baseados em reflectância de superfície e número digital. Mais uma evidência, portanto, da necessidade de se aplicarem a calibração radiométrica e a correção de efeitos atmosféricos aos dados de sensoriamento remoto orbital antes que estes possam ser analisados. CONSIDERAÇÕES FINAIS Os resultados obtidos demonstram a importância incontestável de se aplicar a calibração radiométrica e correção atmosférica para análises de imagens de satélite. Nos gráficos em que se apresentam os valores de reflectância de superfície, reflectância no topo da atmosfera e número digital das bandas 1, 2, 3, 4, 5 e 7, para os alvos área queimada, cultura agrícola, vegetação nativa, copo d’água e solo exposto é visível o comportamento de curva diferente e valores absolutos diferentes para o mesmo pixel, nos diferentes produtos. Obviamente, valores de ND [0, 255] serão largamente diferentes dos valores de reflectância [0, 1] por estarem em escalas diferentes, mas, para além disso, as formas das assinaturas espectrais dos alvos também são amplamente diferentes. De fato, a assinatura esperada para os alvos é completamente descaracterizada quando são empregados valores de ND. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 8 0 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de JENSEN, J. Sensoriamento e remoto do ambiente: uma perspectiva em recursos terrestres. Tradução, prefácio e notas: Jose Carlos Neves Epiphanio (coordenador).São Jose dos Campos - SP: Parêntese, 2009. LEITE, A. P.; SANTOS, G. R.; SANTOS, J. E. O. Análise temporal dos índices de vegetação NDVI e SAVI na estação experimental de Itatinga utilizando imagens Landsat 8. Revista Brasileira de Energias Renováveis. Vol. 6, n. 4, p. 606-623, 2017. MATTHEW, M. W.; ADLER-GOLDEN, S. M.; BERK, A.; FELDE, G.; ANDERSON, G. P.; GORODETZKY, D.; SHIPPERT, M. Atmospheric correction of spectral imagery: evaluation of the FLAASH algorithm with AVIRIS data. In Applied Imagery Pattern Recognition Workshop, 2002. Proceedings. (pp. 157-163). IEEE. NOVO, E. M. L. M. Sensoriamento remoto: princípios e aplicações. São Paulo: Blucher, 2010. PONZONI, F. J.; PINTO, C. T.; LAMPARELLI, R. A. C.; JUNIOR, J. Z.; ANTUNES, M. A. H. Calibração de Sensores Orbitais. São Paulo: Oficina de Textos, 2015. RIBEIRO, B. M. R.; PETRY, F. A.; LIMBERGER, A. R. Análise Temporal de dados NDVI para o município de Toledo PR, obtidos de imagens Landsat 8. Revista Cultivando o Saber, vol. 11, n. 2, p. 149-159, Abr/Jun. 2018. RUMORA, L.; MILER, M.; MEDAK, D. Impact of Various Atmospheric Corrections on Sentinel-2 Land Cover Classification Accuracy Using Machine Learning Classifiers. ISPRS International Journal of Geo-Information, 9(4), 277, 2020. SILVIA, A. M.; MOREIRA, E. B. M. Avaliação espaço-temporal do índice de vegetação (NDVI) no manguezal de Maracaípe – PE, através de imagens do satélite Landsat 5 TM. In: XV Simpósio Brasileiro de Sensoriamento Remoto, Curitiba. Anais do Simpósio Brasileiro de Sensoriamento Remoto, 2011. p. 1570-1575. STEIN, A.; MEER, F. D.; GORTE, B. Spatial Statistics for Remote Sensing. Springer, 2002. STEIN, A.; MEER, F. D.; GORTE, B. Spatial Statistics for Remote Sensing. Springer, 2002. REFERÊNCIAS ANDRADE, C. B.; OLIVEIRA, L. M. M.; OMENA, J. A. M.; VILLAR A. C.; GUSMÃO, L.; RODRIGUES, D. F. B. Avaliação de índices de vegetação e características fisiográficas no Sertão Pernambucano. Revista Brasileira de Meio Ambiente, v.4, n.1. p. 97-107, 2018. ANTUNES, H. M. A.; FREIRE, B. M. R.; BOTELHO, S. A.; TONIOLLI H. L. Correções Atmosféricas de Imagens de Satélites Utilizando o Modelo 6S. In: ANTUNES, H. M. A.; FREIRE, B. M. R.; BOTELHO, S. A.; TONIOLLI H. L. Correções Atmosféricas de Imagens de Satélites Utilizando o Modelo 6S. In: XXI Congresso Brasileiro de Cartografia, Belo Horizonte. Anais do Congresso Brasileiro de Cartografia, 2003. p. 1-6. CHANDER, G. C.; MARKHAM, B. L.; HELDER, D. L. Summary of Current radiometric calibration coefficients for Landsat MSS, TM, ETM+ and EO-1 ALI sensors. Remote Sensing of Environment, v. 113, n. 5, 2009. CHAVEZ, P. S. An improved dark-object subtraction technique for atmospheric scattering correction of multispectral data. Remote sensing of environment, 24(3), 459-479, 1988. GÜRTLER, S.; EPIPHANIO, J. C. N.; LUIZ, A. J. B.; FORMAGGIO, A. R. Planilha Eletrônica para o Cálculo da Reflectância em Imagens TM e ETM+ Landsat. Revista Brasileira de Cartografia, v. 2, n. 57, 2005. G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 8 1 WELTER, André; STATELLA, Thiago; LIMA, Geneci Marques de NOVO, E. M. L. M. Sensoriamento remoto: princípios e aplicações. São Paulo: Blucher, 2010. PONZONI, F. J.; PINTO, C. T.; LAMPARELLI, R. A. C.; JUNIOR, J. Z.; ANTUNES, M. A. H. Calibração de Sensores Orbitais. São Paulo: Oficina de Textos, 2015. Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital Distorções causadas pela falta de calibração radiométrica e efeitos atmosféricos na análise de dados de sensoriamento remoto orbital USGS – United States Geological Survey. LANDSAT 4-7 Surface Reflectance (LEDAPS) Product Guide, version 2. Relatório Técnico. 2019a. USGS – United States Geological Survey. LANDSAT 4-7 Surface Reflectance (LEDAPS) Product Guide, version 2. Relatório Técnico. 2019a. USGS – United States Geological Survey. LANDSAT 7 Data Users Handbook, version 2. Relatório Técnico. 2019b. VELASCO, G. N.; POLIZEL, J. L.; COLTRI, P. P.; LIMA, A. M. L. P.; FILHO, D. F. S. Aplicação do Índice de Vegetação NDVI (Normalized Diference Vegetation G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 8 2 Index) em Imagens de Alta Resolução no Município de São Paulo e suas Limitações. Revista da Sociedade Brasileira de Arborização Urbana, vol. 2, n. 3, 2007. VERMOTE, E. F.; TANRE, D.; DEUZE, J. L.; HERMAN, M. & MORCRETTE, J. J. Second Simulation of the Satellite Signal in the Solar Spectrum, 6S: an overview. IEEE Trans. Geosc. and Remote Sens. vol. 35, n. 3, p. 675- 686, 1997. ZANZARINI, F. V.; PISSARRA, T. C. T.; BRANDÃO, F. J. C.; TEIXEIRA, D. D. B. Correlação Espacial do índice de vegetação (NDVI) de imagem Landsat/ETM+ com atributos do solo. Revista Brasileira de Engenharia Agrícola e Ambiental, vol. 17, n. 6, p. 608-614, 2013. Recebido em 12 de Agosto de 2020 Aceito em 16 de Dezembro de 2020 G E O G R A F I A , V . 4 5 , N . 2 , j u l . / d e z . 2 0 2 0 . 3 8 3	31,424
https://openalex.org/W4387186845_8	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Latin	Spoken	8,614	19,661	Iuda, te laudabunt fratres tui; + manus tua in cervicibus inimicorum tuorum; * adorabunt te filii patris tui. Catulus leonis Iuda: * a praeda, fili mi, ascendisti; requiescens accubuit ut leo * et quasi leaena; quis suscitabit eum? Non auferetur sceptrum de Iuda * et baculus ducis de pedibus eius, donec veniat ille, cuius est, * et cui erit oboedientia gentium; ligans ad vineam pullum suum * et ad vitem filium asinae suae lavabit in vino stolam suam * et in sanguine uvae pallium suum; [267] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 393 nigriores sunt oculi eius vino * et dentes eius lacte candidiores. Zabulon in litore maris habitabit + et in statione navium, * pertingens usque ad Sidonem. Issachar asinus fortis, * accubans inter caulas vidit requiem quod esset bonam, * et terram quod optima; et supposuit humerum suum ad portandum * factusque est tributis serviens. Dan iudicabit populum suum * sicut una tribuum Israel. Fiat Dan coluber in via, cerastes in semita, + mordens calcanea equi, ut cadat ascensor retro. * Salutare tuum exspectabo, Domine! Gad, latrones aggredientur eum, * ipse autem aggredietur calcaneum eorum. Aser, pinguis panis eius, * et praebebit delicias regales. Nephtali cerva emissa, * dans cornua pulchra. Arbor fructifera Ioseph, + arbor fructifera super fontem: * rami transcendunt murum. Sed exasperaverunt eum et iurgati sunt, * et adversati sunt illi habentes iacula. Et confractus est arcus eorum. * et dissoluti sunt nervi brachiorum eorum per manus Potentis Iacob, * per nomen Pastoris, Lapidis Israel. Deus patris tui erit adiutor tuus, * et Omnipotens bendicet tibi benedictionibus caeli desuper, + benedictionibus abyssi iacentis deorsum, * bendictionibus uberum et vulvae. Benedictiones patris tui confortatae sunt + super benedictiones montium aeternorum, * desiderium collium antiquorum; fiant in capite Ioseph * et in vertice nazaraei inter fratres suos. Beniamin lupus rapax; + mane comedet praedam * et vespere dividet spolia. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Deus patris nostri erit adiutor noster, et omnipotens benedicet nobis. 394 GABRIEL RAMIS MIQUEL [268] Oratio de cantico (Br Goth 342) Appareat quaesumus, Domine, mentibus nostris agnitio tuae divinitatis, quae benedictionis habere gratiam praebeat nobis; ut proficiamus et crescamus iugiter in bonis multiplicibus incrementis. Deus patris nostri erit adiutor noster, et omnipotens benedicet nobis. R/ Amen. Adiuvante tua dextera, Deus noster, qui vivis et regnas in saecula saeculorum. R/ Amen. Psalmus matutinarius (Br Goth 342; T35.2, 327; AL 177) (Ps 75, 5.2) Illuminans tu mirabilis, a montibus aeternis. V/ Notus in Iudaea Deus, in Israel magnum nomen eius. R/ A montibus aeternis. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ A montibus aeternis. Psalmi laudate 148-150 Antiphona (Br Goth 342; T35.2, 338; AL 241) (Cfr. Ps 149, 3; cfr. Ps 150, 3-4) In psalterio et cithara hymnum dicamus; in choro laus resonet Domino in chordis et organo. (Ut in ordinario in officio feriali) Ant. In psalterio et cithara hymnum dicamus; in choro laus resonet Domino in chordis et organo. Lectio (Br Goth 342-343) (Gn 46,28 - 47,12) Lectio libri Genesis. R/ Deo gratias. In illis diebus: Missit autem Iudam ante se Ioseph […] et alebat eos omnemque domum patris sui praebens cibaria singulis. R/ Amen. [269] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 395 Hymnus (Br Goth 197; T35.2, 330, 64) Deus aeterni luminis candor inenarrabilis, venturus diei iudex, qui mentis occulta vides. Tu mundi constitutor es Tu sempiterno throno sedes, Iudex, ex alto Humilis, venisti pati pro nobis. Tu regnum caelorum tenes, et totus in Verbo tu es, per Filium cuncta regis Sancto Spiritui fons es. Tu Sabaoth omnipotens, Tu Hosanna summi culminis, tibi laus est mirabilis, Tu es prima Anastasis. Trinum nomen alta vides, unum per omnia potes, mirumque per signum crucis tu rector immensae lucis. Tu fidei auditor es, et humiles Tu respicis, tibi altum vides thronum, tibi divinus est honor. Christoque, aeterno Deo Patri, cum Sancto Spiritu, In te solvamus munera a saeculis in saecula. Amen. Versus (AL 158, 170) (Ps 62, 7-8) V/ In matutinis meditabor in te. R/ Quia factus es adiutor meus. Supplicatio (Br Goth 259) Oremus. Redemptorem mundi, Dominum nostrum Iesum Christum, cum omni supplicatione rogemus; ut nos, gratia ipsius, et per intercessionem sanctae Mariae Virginis, et Genetricis suae, ut sancto tempore quadragesimae, nobis remissionem peccatorum, et pacem nobis donare dignetur. R/ Praesta aeterne omnipotens Deus. Kyrie eleison. R/ Christe eleison, Kyrie eleison. 396 GABRIEL RAMIS MIQUEL Completuria (Br Goth 281, 264; T35.2, 331, 4; LOPs 469) Domine, apud quem est salus plena iustitiae et perfectio beatitudinis incorruptae: da nobis, ut in legis tuae meditatione diei ac noctis tempora transeamus, et ab impiorum consilio, atque a peccatorum via, vel a pestilentiae cathedra declinemus; quo velut lignum fructiferum, aquarum decursibus enutritum, et fructibus repleamur, et gratia decoremur. R/ Amen. Per misericordiam ipsius Dei nostri, qui est benedictus et vivit et omnia regit, in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) Benedictio (Br Goth 341) Diac. Humiliate vos benedictioni. Presb. Dominus sit semper vobiscum. R/ Et cum spiritu tuo. Omnipotentis Dei vos gratia protegat, famemque vestram spirituali cibo reficiat. R/ Amen. Sitim quoque vestram iustitiae fluminibus irriget, atque novum dulcedinis suae animis vestris infundat saporem. R/ Amen. Ut tam carne contriti quam spiritu sanctificati, ab eo mereamini caelesti dono beatificari. R/ Amen. Per misericordiam ipsius Dei nostri, qui vivit et omnia regit in saecula saeculorum. R/ Amen. [270] [271] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 397 Psallendum (Br Goth 343; T35.2, 332) (Ps 118, 43.41) Ne auferas de ore meo verbum veritatis, Domine; quia in iudiciis tuis speravi. V/ Et veniat super me misericordia tua, Domine; salutare tuum et eloquium tuum. R/ Quia in iudiciis tuis speravi. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Quia in iudiciis tuis speravi. Oratio (Br Goth 343, 330; T35.2, 1498; LOPs 284) Tua super nos, Domine, misericordia, te dum rogamus, adveniat; tuumque salutare cor nostrum erudiat; eloquii eruditio mentem nostram enutriat; ut qui in tuis speramus sermonibus, tuis per omnia consolemur virtutibus; ac dum spes nostra in te dirigitur, spei ipsius a te nobis petitio concedatur. R/ Amen. Praestante ipsius misericordia Dei nostri, qui in Trinitate, unus Deus, vivit et gloriatur in saecula saeculorum. R/ Amen. Conclusio (Ut in ordinario in officio feriali) Ad tertiam Lectio (Br Goth 343) (Prv 11, 23 – 12, 3) Lectio libri Proverbiorum Salomonis. R/ Deo gratias. Fili: Desiderium iustorum omne bonum est […] et radix iustorum non commovebitur. R/ Amen. Lectio (Br Goth 343-344) (Gn 47, 15-26) Lectio libri Genesis. R/ Deo gratias. 398 GABRIEL RAMIS MIQUEL [272] In illis diebus: Venit cuncta Aegyptus ad Iosep […] et factum est a Ioseph in legem absque terra sacerdotali, quae libera ab hac conditione est. R/ Amen. Psalmus 44 Antiphona (Br Goth 344; T35.2, 333) (Ps 44, 10) Populi confitebuntur tibi in aeternum, et in saeculum saeculi. Eructavit cor meum verbum bonum, * dico ego opera mea regi. Lingua mea calamus scribae * velociter scribentis. Speciosus forma es prae filiis hominum, + diffusa est gratia in labiis tuis, * propterea benedixit te Deus in aeternum. Accingere gladio tuo super femur tuum, potentissime, * magnificentia tua et ornatu tuo. Et ornatu tuo procede, currum ascende + propter veritatem et mansuetudinem et iustitiam. * Et doceat te mirabilia dextera tua: sagittae tuae acutae populi sub te cadent * in corda inimicorum regis. Sedes tua, Deus, in saeculum saeculi; * sceptrum aequitatis sceptrum regni tui. Dilexisti iustitiam et odisti iniquitatem, * propterea unxit te Deus, Deus tuus, oleo laetitiae prae consortibus tuis. Myrrha et aloe et casia omnia vestimenta tua; * e domibus eburneis chordae delectant te. Filiae regum in pretiosis tuis; * astitit regina a dextris tuis ornata auro ex Ophir. Audi, filia, et vide et inclina aurem tuam * et obliviscere populum tuum et domum patris tui; et concupiscet rex speciem tuam. * Quoniam ipse est dominus tuus, et adora eum. Filia Tyri cum muneribus; * vultum tuum deprecabuntur divites plebis. Gloriosa nimis filia regis intrinsecus, * texturis aureis circumamicta. In vestibus variegatis adducetur regi; * virgines post eam, proximae eius, afferuntur tibi. Afferuntur in laetitia et exsultatione, * adducuntur in domum regis. [273] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 399 Pro patribus tuis erunt tibi filii; * constitues eos principes super omnem terram. Memor ero nominis tui * in omni generatione et generatione; propterea populi confitebuntur tibi in aeternum * et in saeculum saeculi. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Populi confitebuntur tibi in aeternum, et in saeculum saeculi. Oratio (Br Goth 344, 284; T35.2, 334, 80, 569) Fac nos, Domine, diligere iustitiam et odire iniquitatem, ac per speciem, tuam pulchritudinem quaerentes, deducat nos mirabiliter dextera tua; et, mortificato in nobis incentivo fornicationis, accinge lumbos nostros gladio castitatis. R/ Amen. Per ineffabilem bonitatem tuam, Deus noster, qui vivis et cuncta dominaris in saecula saeculorum. R/ Amen. Psalmus 45 Antiphona (Br Goth 344; T35.2, 335) (Ps 45, 8) Dominus virtutum nobiscum, susceptor noster Deus Israel. Deus est nobis refugium et virtus, * adiutorium in tribulationibus inventus est nimis. Propterea non timebimus, dum turbabitur terra, * et transferentur montes in cor maris. Fremant et intumescant aquae eius, * conturbentur montes in elatione eius. Fluminis rivi laetificant civitatem Dei, * sancta tabernacula Altissimi. Deus in medio eius, non commovebitur; * adiuvabit eam Deus mane diluculo. Fremuerunt gentes, commota sunt regna; * dedit vocem suam, liquefacta est terra. Dominus virtutum nobiscum, * refugium nobis Deus Iacob. Venite et videte opera Domini, * quae posuit prodigia super terram. Auferet bella usque ad finem terrae, * arcum conteret et confringet arma et scuta comburet igne. 400 GABRIEL RAMIS MIQUEL [274] Vacate et videte quoniam ego sum Deus: * exaltabor in gentibus et exaltabor in terra. Dominus virtutum nobiscum, * refugium nobis Deus Iacob. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant Dominus virtutum nobiscum, susceptor noster Deus Israel. Oratio (Br Goth 344, 225; T35.2, 336; MPs 16; LOPs 358) Deus, virtus et refugium nostrum: esto nobis adiutor in tribulationibus; ut in nullo nostra metuat conscientia, cum te iudicante conturbata fuerit terra; fluminis tui impetus, tamquam in sanctis habitans, laetificat civitatem; atque in eo sumat ex vultu tuo adiutorium, cum Sancti Spiritus, te largiente, perceperit donum. R/ Amen. Per ineffabilia mirabilia sua, Deus noster, qui vivit et regnat in saecula saeculorum. R/ Amen. Psalmus 53 Antiphona (Br Goth 344; T35.2, 337; cfr. AL 182) (Cfr. Ps 53, 3-4a) Deus, in nomine tuo salvos nos fac, et in virtute tua libera nos. Deus, exaudi orationem nostram. Deus, in nomine tuo salvum me fac * et in virtute tua iudica me. Deus, exaudi orationem meam, * auribus percipe verba oris mei! Quoniam superbi insurrexerunt adversum me, + et fortes quaesierunt animam meam * et non proposuerunt Deum ante conspectum suum. Ecce enim Deus adiuvat me, * et Dominus susceptor est animae meae. Converte mala super inimicos meos * et in veritate tua disperde illos. Voluntarie sacrificabo tibi, * confitebor nomini tuo, Domine, quoniam bonum est; quoniam ex omni tribulatione eripuit me, * et super inimicos meos despexit oculus meus. [275] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 401 Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Deus, in nomine tuo salvos nos fac, et in virtute tua libera nos. Deus, exaudi orationem nostram. Responsorium (Br Goth 344-345; T35.2, 338; cfr. AL 190) (Cfr. Is 33, 2; cfr. Ps 101, 2) Domine, miserere nobis, te enim expectavimus tota die; esto brachium nostrum in fortitudine, et salus nostra in tempore tribulationis. V/ Domine, exaudi orationem nostram, et clamor noster ad te perveniat. R/ In tempore tribulationis. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ In tempore tribulationis. Preces (Br Goth 345) Diac. Flectamus genua. Presb. Deprecemur Dominum. R/ Kyrie eleison. Christe eleison. Kyrie eleison. Diac. Erigite vos. Dicamus omnes: miserere, Domine, et exaudi nos, quia pius es. V/ Ex toto corde et ex tota mente orantes ad te, exaudi, Domine. R/ Quia pius es. V/ Qui habitas in caelis et humilia respicis, paenitentibus dona indulgentiam. R/ Quia pius es. V/ Civitatem hanc, et omnes habitantes in ea, protege, Christe, dextera virtutis tuae. R/ Quia pius es. V/ Remissionem peccatorum, et emmendationem morum, concede, Domine. R/ Quia pius es. 402 GABRIEL RAMIS MIQUEL [276] Hymnus (Br Goth 270) Frenentur ergo corporum cupidines, detersa et intus emicet prudentia: sic excitato perspicax acumine, liberque flatu laxiore spiritus rerum Parentem rectius precabitur. Elias tali crevit observantia vetus sacerdos, ruris hospes aridi: fragore ab omni quem remotum et segregem sprevisse tradunt criminum frequentiam, casto fruentem syrtium silentio. Honor, potestas, plenitudo gratiae Deo perenni, Trinitati simplici, aeterna, cuius est beata gloria, replet globi, rerumque cuncta machina, tenetque regnum perpetim per saecula. Amen. Supplicatio (Ut in ordinario in officio feriali) Completuria (Br Goth 345; T35.2, 341; LOPs 371) Salva nos, Domine, in tuo nomine, et libera in virtute; educ de tribulatione, et reple iucunditate; ne a salute nostra praesidium retrahas, ne quominus liberemur virtutem impendas; ut dum nobis salus tuo praestatur ex munere, tibi gloria rependatur in laudem, et nobis libertas contingat in requiem. R/ Amen. Praestante ipsius misericordia Dei nostri, qui in Trinitate, unus Deus, vivit et gloriatur in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) Benedictio (Ut in ordinario in officio feriali) Conclusio (Ut in ordinario in officio feriali) [277] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 403 Ad sextam Lectio (Br Goth 345) (Is 63, 15-64, 8a) Lectio libri Isaiae Prophetae. R/ Deo gratias. Attende de caelo et vide […] Ne irascaris, Domine, nimis et ne ultra memineris iniquiatatis. R/ Amen. Psalmus 57 Antiphona (Br Goth 346; T35.2, 342) (Ps 57, 2b) Recte iudicate filii hominum. Numquid vere, potentes, iustitiam loquimini, * recte iudicatis filios hominum? Etenim in corde iniquitates operamini, * in terra violentiam manus vestrae concinnant. Alienati sunt peccatores ab utero; * erraverunt a ventre, qui loquuntur falsa. Venenum illis in similitudinem serpentis, * sicut aspidis surdae et obturantis aures suas, quae non exaudiet vocem incantantium * et venefici incantantis sapienter. Deus, contere dentes eorum in ore ipsorum; * molas leonum confringe, Domine. Diffluant tamquam aqua decurrens, * sicut fenum conculcatum arescant. Sicut limax, quae tabescens transit, * sicut abortivum mulieris, quod non vidit solem. Priusquam sentiant ollae vestrae rhamnum, * sicut viventes, sicut ardor irae absorbet eos. Laetabitur iustus, cum viderit vindictam, * pedes suos lavabit in sanguine peccatoris. Et dicet homo:“Utique est fructus iusto, * utique est Deus iudicans eos in terra”. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Recte iudicate filii hominum. 404 GABRIEL RAMIS MIQUEL [278] Oratio (Br Goth 346, 421, 1032; T35.2, 345; LOPs 525) Deus, cuius sunt eloquia casta, et iudicia recta: tribue nobis casto ore verba tua depromere, et corde purissimo iudiciorum tuorum mandata servare; extende itaque arcum tuum ob defensionem mostram; et ad infirmanda inimicorum consilia, spiritualia dirige tela; molasque confringe leonum, quae tuum demoliri nituntur sceptrum; ut in te laetetur copia redemptorum, qui es corona iustorum; tibique decantemus psalmum laetitiae, cuius munere pervenitur ad fructum iustitiae. R/ Amen. Praestante ipsius divinitatis clementia, qui in Trinitate, unus Deus, vivit et regnat in saecula saeculorum. R/ Amen. Psalmus 125 Antipohna (Br Goth 346; T35.2, 343) (Ps 125, 4) Converte, domine, captivitatem nostram, sicut torrens in austro. In convertendo Dominus captivitatem Sion, * facti sumus quasi somniantes. Tunc repletum est gaudio os nostrum, * et lingua nostra exsultatione. Tunc dicebant inter gentes: * “Magnificavit Dominus facere cum eis”. Magnificavit Dominus facere nobiscum; * facti sumus laetantes. Converte, Domine, captivitatem nostram, * sicut torrentes in austro. Qui seminant in lacrimis, * in exsultatione metent. Euntes ibant et flebant * semen spargendum portantes; venientes autem venient in exsultatione * portantes manipulos suos. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Converte, domine, captivitatem nostram, sicut torrens in austro. [279] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 405 Oratio (Br Goth 346, 166-167, 372; LOPs 452) Convertere, Domine, captivitatem nostram, ab illecebrosis inimici seductionibus redimendo; et reple consolatione indulgentiae, quos desolatos prospicis ob admissionem nequitiae; da nobis quaesumus, contriti cordis humilitatem quaerere, et inquinamenta, quae nos polluunt, fluentis lacrimarum abluere; sic etiam confessionis remedia seramus in lacrimis, ut indulgentiae fructus metamus in gaudiis. R/ Amen. Praestante sua misericordia, qui in Trinitate, unus Deus, vivit et regnat in saecula saeculorum. R/ Amen. Psalmus 123 Antiphona (Br Goth 346; T35.2, 344) (Ps 123, 8) Adiutorium nostrum in nomine Domini, qui fecit caelum et terram. Nisi quia Dominus erat in nobis, * dicat nunc Israel, nisi quia Dominus erat in nobis, * cum exsurgerent homines in nos: forte vivos deglutissent nos, * cum irasceretur furor eorum in nos. Forsitan aqua absorbuisset nos, * torrens pertransisset animam nostram; forsitan pertransissent animam nostram * aquae intumescentes. Benedictus Dominus, * qui non dedit nos in direptionem dentibus eorum. Anima nostra sicut passer erepta est * de laqueo venantium: laqueus contritus est, * et nos erepti sumus. Adiutorium nostrum in nomine Domini, * qui fecit caelum et terram. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Adiutorium nostrum in nomine Domini, qui fecit caelum et terram. 406 GABRIEL RAMIS MIQUEL [280] Responsorium (Br Goth 346; T35.2, 347; AL 337) (Cfr. 2Reg 19, 15; cfr. Ps 66, 5; cfr. Ps 9, 10.35; Ps 79, 2) Qui sedes super thronum et iudicas aequitatem; esto refugium pauperum in tribulatione, quia tu solus laborem et dolorem consideras. V/ Qui regis Israel, intende; qui deducis velut ovem Ioseph, qui sedes super cherubim. R/ Esto refugium pauperum in tribulatione, quia tu solus laborem et dolorem consideras. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Esto refugium pauperum in tribulatione, quia tu solus laborem et dolorem consideras. Preces (Br Goth 346) Diac. Flectamus genua. Presb. Deprecemur Dominum. R/ Kyrie eleison. Christe eleison. Kyrie eleison. Diac. Erigite vos. Dicamus omnes: propitiare populo tuo, Christe, et miserere. V/ Intende nostris tandem precibus, intende iam propitius. R/ Et miserere. V/ Nos peccavimus, nos inique egimus. R/ Et miserere. V/ Averte iram tuam, et dona indulgentiam. R/ Et miserere. V/ A peccatis ac delicti nostris purifica nos, Domine. R/ Et miserere. [281] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 407 Hymnus (Br Goth 271-272) Ionam prophetam mitis ultor excitat, poenae imminentis iret ut praenuncius; sed nosset ille cum minacem iudicem servare malle, quam ferire, ac plectere, tectam latenter vertit Tharsos fugam. Iussus perire solus e cunctis reus, cuius voluta crimen urna expresserat, praeceps rotatur, et profundo immergitur; exceptus inde belluinis faucibus, alvi capacis vivus hauritur specu. Celsam paratis pontibus scandit ratem, udo revincta fune puppis solvitur; itur per altum, fit procelosum mare; tunc causa tanti quaeritur periculi, sors in fugacem missa vatem decidit. Honor, potestas, plenitudo gratiae Deo perenni, Trinitati simplici, aeterna, cuius est beata gloria, replet globi, rerumque cuncta machina, tenetque regnum perpetim per saecula. Amen. Supplicatio (Ut in ordinario in officio feriali) Oratio (Br Goth 346, 338-339; T35.2, 349, 296; LOPs 187) Adiutorium nostrum in nomine tuo est, Domine; da nobis odire mundi illecebram, et venantium voluptatum praecavere muscipulam; ut ipse insidiantem perimas, ipse confidentem munias atque defendas. R/ Amen. Concedente clementia sua, qui in Trinitate, unus Deus, vivit et regnat in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) 408 GABRIEL RAMIS MIQUEL [282] Benedictio (Ut in ordinario in officio feriali) Conclusio (Ut in ordinario in officio feriali) Ad nonam Lectio (Br Goth 346) (Prv 23, 22-25) Lectio libri Proverbiorum Salomonis. R/ Deo gratias. Fili, audi patrem tuum […] et exultet quae genuit te. R/ Amen. Lectio (Br Goth 346-348) (Gn 47, 27 – 48, 22) Lectio libri Genesis. R/ Deo gratias. In illis diebus: Habitavit ergo Israel in Aegypto […] quam tuli de manu Amorrei in gladio et arcu meo. R/ Amen. Psalmus 59 Antiphona (Br Goth 348; T35.2, 350) (Cfr. Ps 59, 12b) Da nobis auxilium, Deus, in tribulatione. Deus, reppulisti nos, destruxisti nos. * Iratus es. Convertere ad nos! Concussisti terram, confregisti eam; * sana contritiones eius, quia commota est. Ostendisti populo tuo dura, * potasti nos vino vertiginis. Dedisti metuentibus te signum, * ut fugiant a facie arcus. Ut liberentur dilecti tui, * salvos fac dextera tua et exaudi nos. Deus locutus est in sancto suo: + “Laetabor et partibor Sichimam * et convallem Succoth metibor. Meus est Galaad, et meus est Manasses, * et Ephraim fortitudo capitis mei. Iuda sceptrum meum, * Moab olla lavacri mei. Super Idumaeam extendam calceamentum meum, * super Philistaeam vociferabor”. [283] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 409 Quis adducet me in civitatem munitam? * Quis deducet me usque in Idumaeam? Nonne tu, Deus, qui reppulisti nos; * et non egredieris, Deus, in virtutibus nostris? Da nobis auxilium de tribulatione, * quia vana salus hominis. In Deo faciemus virtutem, * et ipse conculcabit tribulantes nos. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Da nobis auxilium, Deus, in tribulatione. Oratio (Br Goth 348, 401; T35.2, 351, 594; LOPs 71) Da nobis, Domine, auxilium in tribulatione, quia vana est salus ab homine; in te enim faciemus virtutem, qui et confortas famulos, et ad nihil redigis inimicos. R/ Amen. Auxiliante ipsius misericordia Domini nostri, qui omnia regit in saecula saeculorum. R/ Amen. Psalmus 65 Antiphona (T35.2, 352) (Ps 65, 4) Omnis terra adoret te, et psallat tibi, Domine. Iubilate Deo, omnis terra, + psalmum dicite gloriae nominis eius, * glorificate laudem eius. Dicite Deo: “Quam terribilia sunt opera tua. * Prae multitudine virtutis tuae blandientur tibi inimici tui. Omnis terra adoret te et psallat tibi, * psalmum dicat nomini tuo”. Venite et videte opera Dei, * terribilis in adinventionibus super filios hominum. Convertit mare in aridam, + et in flumine pertransibunt pede; * ibi laetabimur in ipso. Qui dominatur in virtute sua in aeternum, + oculi eius super gentes respiciunt; * rebelles non exaltentur in semetipsis. Benedicite, gentes, Deum nostrum * et auditam facite vocem laudis eius; qui posuit animam nostram ad vitam * et non dedit in commotionem pedes nostros. 410 GABRIEL RAMIS MIQUEL [284] Quoniam probasti nos, Deus; * igne nos examinasti, sicut examinatur argentum. Induxisti nos in laqueum, * posuisti tribulationes in dorso nostro. Imposuisti homines super capita nostra, + transivimus per ignem et aquam, * et eduxisti nos in refrigerium. Introibo in domum tuam in holocaustis;* reddam tibi vota mea, quae protulerunt labia mea, * et locutum est os meum in tribulatione mea. Holocausta medullata offeram tibi cum incenso arietum, * offeram tibi boves cum hircis. Venite, audite, et narrabo, omnes, qui timetis Deum, * quanta fecit animae meae. Ad ipsum ore meo clamavi * et exaltavi in lingua mea. Iniquitatem si aspexi in corde meo, * non exaudiet Dominus. Propterea exaudivit Deus, * attendit voci deprecationis meae. Benedictus Deus, qui non amovit orationem meam * et misericordiam suam a me. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Omnis terra adoret te, et psallat tibi, Domine. Oratio (Br Goth 348, 165, 670-671; T35.2, 353: LOPs 78) Omnis terra adoret te, Domine, et tibi psallat vitae participio incorruptae; ut sicut sunt omnia tuo formata opificio, ita cuncta tuo pariter subdantur imperio. R/ Amen. Auxiliante misericordia divinitatis suae, qui in Trinitate, unus Deus, vivit et regnat in saecula saeculorum. R/ Amen. Psalmus 69 Antiphona (Br Goth 348; T35.2, 354) (Cfr. Ps 69, 2) In adiutorium nostrum intende, Domine; ad adiuvando nos festina. [285] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 411 Deus, in adiutorium meum intende; * Domine, ad adiuvandum me festina. Confundantur et revereantur, * qui quaerunt animam meam. Avertantur retrorsum et erubescant, * qui volunt mihi mala. Convertantur propter confusionem suam, * qui dicunt mihi: “Euge, euge”. Exsultent et laetentur in te omnes, qui quaerunt te; + et dicant semper: “Magnificetur Deus”, * qui diligunt salutare tuum. Ego vero egenus et pauper sum; * Deus, ad me festina. Adiutor meus et liberator meus es tu; * Domine, ne moreris. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. In adiutorium nostrum intende, Domine; ad adiuvando nos festina. Hymnus (Br Goth 273) Mirator hostis posse limum tabidum tantum laboris sustinere, ac perpeti. Explorat arte sciscitator callida, Deus ne membris sit receptus terreis: sed, increpata fraude, post tergum ruit. Hoc est, quod atri livor hostis invidet mundi, polique quod gubernator probat, altaris aram quod facit placabilem, quod dormientis excitat cordis fidem, quod limat aegram corporis libidinem. Hoc nos sequamur, quisque nunc pro viribus, quod consecrati tu Magister dogmatis tuis dedisti, Christe sectatoribus: ut, cum vorandi vicerit libidinem, late triumphet imperator spiritus. Honor, potestas, plenitudo gratiae Deo perenni, Trinitati simplici, aeterna, cuius est beata gloria, replet globi, rerumque cuncta machina, tenetque regnum perpetim per saecula. Amen. Supplicatio (Ut in ordinario in officio feriali) Completuria (Br Goth 348, 712; T35.2, 356; LOPs 254) In adiutorium nostrum intende, Domine, nosque ad adiuvandum festina; 412 GABRIEL RAMIS MIQUEL [286] nihil in bonis operibus conatus nostri te desistente proficiunt, a quo gressus hominis diriguntur: tu es enim per quem nostra roboratur fragilitas, et expiatur iniquitas, tribuitur iustitia redditurque corona. R/ Amen. Ipso auxiliante Domino, qui cum Patre et Spiritu Sancto, unus Deus, gloriatur in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) Benedictio (Ut in ordinario in officio feriali) Conclusio (Ut in ordinario in officio feriali) Ad vesperum Oblatio luminis (Ut in ordinario in officio feriali) Vespertinum (Br Goth 348; T35.2, 357) (Ps 26, 1) Illuminatio mea et salus mea Dominus. Psalmus 128 Antiphona (Br Goth 348; T35.2, 358) (Ps 128, 8) Benedictio Domini super vos, benedicimus in nomine Domini. Saepe expugnaverunt me a iuventute mea, * dicat nunc Israel, saepe expugnaverunt me a iuventute mea, * etenim non potuerunt adversum me. [287] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 413 Supra dorsum meum araverunt aratores, * prolongaverunt sulcos suos. Dominus autem iustus * concidit cervices peccatorum. Confundantur et convertantur retrorsum * omnes, qui oderunt Sion. Fiant sicut fenum tectorum, * quod, priusquam evellatur, exaruit; de quo non implevit manum suam, qui metit, * et sinum suum, qui manipulos colligit. Et non dixerunt, qui praeteribant: + “Benedictio Domini super vos, * benedicimus vobis in nomine Domini”. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Benedictio Domini super vos, benedicimus in nomine Domini. Alleluiaticum (Br Goth 248; T35.2, 359) (Ps 118, 37-38) Averte oculos meos ne videant vanitatem, et in via tua vivifica me, Domine. V/ Statue servo tuo eloquium tuum, in itinere tuo. R/ Et in via tua vivifica me, Domine. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Et in via tua vivifica me, Domine. Hymnus (Br Goth 287; T35.2, 360, 93, 612) Christe, lumen perpetuum astrorum, fulgor siderum, noctem nostram illumina, conserva pura pectora. Custodi corda omnium fideliumque maxima protectione propia tua, Christe, potentia. Nullis incedat laqueis nullis gravetur hostibus, nec maculetur corpus lascivia per soporem. Ut gratuletur anima vidisse lucem prosperam, implesse vota debita reddenda tibi, Trinitas. Deo Patri sit gloria, eiusque soli Filio cum Spiritu Paraclito in sempiterna saecula. Amen. 414 GABRIEL RAMIS MIQUEL [288] Versus (AL 154) (Ps 16, 5) V/ Perfice gressus meos in semitis tuis. R/ Ut non moveantur vestigia mea. Supplicatio (Br Goth 259) Oremus. Redemptorem mundi, Dominum nostrum Iesum Christum, cum omni supplicatione rogemus; ut nos, gratia ipsius, et per intercessionem sanctae Mariae Virginis, et Genetricis suae, ut sancto tempore quadragesimae, nobis remissionem peccatorum, et pacem nobis donare dignetur. R/ Praesta aeterne omnipotens Deus. Kyrie eleison. R/ Christe eleison, Kyrie eleison. Completuria (Br Got 341; T35.2, 361, 182, 313) Servis tuis haec est, Domine, vita sub spe vitae in hoc saeculo laborare, et per illuminationem gratiae tuae iugiter ad te ieiuniis et orationibus vigilare. Consecra ergo, quaesumus, ieiunium nostrum, et repelle ab animis nostris nostrorum sarcinam delictorum; quatenus hic superato diabolo, mereamur cum omnibus sanctis tuis congaudere in regno perpetuo. R/ Amen. Per misericordiam tuam, Deus noster, qui es benedictus, et vivis et omnia regis in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) [289] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 415 Benedictio (Br Goth 341) Diac. Humiliate vos benedictioni. Presb. Dominus sit semper vobiscum. R/ Et cum spiritu tuo. Omnipotentis Dei vos gratia protegat, famemque vestram spirituali cibo reficiat. R/ Amen. Sitim quoque vestram iustitiae fluminibus irriget, atque novum dulcedinis suae animis vestris infundat saporem. R/ Amen. Ut tam carne contriti quam spiritu sanctificati, ab eo mereamini caelesti dono beatificari. R/ Amen. Per misericordiam ipsius Dei nostri, qui vivit et omnia regit in saecula saeculorum. R/ Amen. Psallendum (Br Goth 343; T35.2, 332) (Ps 118, 43.41) Ne auferas de ore meo verbum veritatis, Domine; quia in iudiciis tuis speravi. V/ Et veniat super me misericordia tua, Domine; salutare tuum et eloquium tuum. R/ Quia in iudiciis tuis speravi. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Quia in iudiciis tuis speravi. Oratio (Br Goth 343, 330; T35.2, 1498; LOPs 284) Tua super nos, Domine, misericordia, te, dum rogamus adveniat; tuumque salutare cor nostrum erudiat: eloquii eruditio mentem nostram enutriat; ut qui in tuis speramus sermonibus, tuis per omnia consolemur virtutibus; ac dum spes nostra in te dirigitur, spei ipsius a te nobis petitio concedatur. R/ Amen. Praestante ipsius misericordia Dei nostri, qui in Trinitate, unus Deus, vivit et gloriatur in saecula saeculorum. R/ Amen. 416 GABRIEL RAMIS MIQUEL [290] Conclusio (Ut in ordinario in officio feriali) Feria IV secundae hebdomadae Quadragesimae Ad matutinum Psalmus III Antiphona (Br Goth 288; T35.2, 362) (Ps 3, 4b-5) Gloria mea et exaltans caput meum. Voce mea ad Dominum clamavi, et exaudivit me de monte sancto suo. (Ut in ordinario in officio feriali) Ant. Gloria mea et exaltans caput meum. Voce mea ad Dominum clamavi, et exaudivit me de monte sancto suo. Collecta de psalmo III (Br Goth 288, 467; T35.2, 50, 185, 567, 878, 1044; LOS 543; S6, 19; OV 1188; LOPs 321) Gloria nostra, Deus noster: da nobis ut in te sine fine gloriemur; quosque fecisti de capitis exaltatione sublimes, quaesumus ut efficias de totius corporis consummata salute securos; elevet abiectionem nostram tua magna celsitudo; ut qui humilitate tua a terrenis erigimur, exaltatione quoque tua ad caelestia pervehi mereamur. R/ Amen. Per misericordiam ipsius Dei nostri, qui vivit et omnia regit in saecula saeculorum. R/ Amen. Missa Psalmus 136 Antiphona (Br Goth 348; T35.2, 364) (Cfr. Ps 136, 3c) Cantemus Domino de canticis Sion, ut recordetur nostri et liberet nos. [291] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 417 Super flumina Babylonis, illic sedimus et flevimus, * cum recordaremur Sion. In salicibus in medio eius * suspendimus citharas nostras. Quia illic rogaverunt nos, qui captivos duxerunt nos, * verba cantionum, et, qui affligebant nos, laetitiam: * “Cantate nobis de canticis Sion”. Quomodo cantabimus canticum Domini * in terra aliena? Si oblitus fuero tui, Ierusalem, * oblivioni detur dextera mea; adhaereat lingua mea faucibus meis, * si non meminero tui, si non praeposuero Ierusalem * in capite laetitiae meae. Memor esto, Domine, adversus filios Edom * diei Ierusalem; qui dicebant: “Exinanite, exinanite * usque ad fundamentum in ea”. Filia Babylonis devastans, * beatus, qui retribuet tibi retributionem tuam, quam retribuisti nobis; beatus, qui tenebit * et allidet parvulos tuos ad petram. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Cantemus Domino de canticis Sion, ut recordetur nostri et liberet nos. Oratio (Br Goth 348; T35.2, 365; LOPs 199) Peregrinantis ecclesiae tuae, Domine, memor esto, ac dum super flumina Babylonis deflentes sedimus, praetereuntis saeculi impetu non trahamur; sed a praesentis vitae liberati contagio, ad Ierusalem supernam erigatur intentio. R/ Amen. Concedente eius misericordia, qui vivit et regnat Deus in saecula saeculorum. R/ Amen. Psalmus 72 Antiphona (Br Goth 349; T35.2, 366; AL 476) (Cfr. Ps 72, 28) Nobis adhaerere Deo bonum est, ponere in Deo spem nostram. Quam bonus rectis est Deus, * Deus his, qui mundo sunt corde! Mei autem paene moti sunt pedes, * paene effusi sunt gressus mei, 418 GABRIEL RAMIS MIQUEL [292] quia zelavi super gloriantes, * pacem peccatorum videns. Quia non sunt eis impedimenta, * sanus et pinguis est venter eorum. In labore mortalium non sunt * et cum hominibus non flagellantur. Ideo quasi torques est eis superbia, * et tamquam indumentum operuit eos violentia. Prodit quasi ex adipe iniquitas eorum, * erumpunt cogitationes cordis. Subsannaverunt et locuti sunt nequitiam, * iniquitatem ab excelso locuti sunt. Posuerunt in caelo os suum, * et lingua eorum transivit in terra. Ideo in alto sedent, * et aquae plenae non pervenient ad eos. Et dixerunt: “Quomodo scit Deus, * et si est scientia in Excelso?”. Ecce ipsi peccatores et abundantes in saeculo * multiplicaverunt divitias. Et dixi: “Ergo sine causa mundavi cor meum * et lavi in innocentia manus meas; et fui flagellatus tota die, * et castigatio mea in matutinis”. Si dixissem: “Loquar ut illi”, * ecce generationem filiorum tuorum prodidissem. Et cogitabam, ut cognoscerem hoc; * labor erat in oculis meis, donec intravi in sanctuarium Dei * et intellexi novissima eorum. Verumtamen in lubrico posuisti eos, * deiecisti eos in ruinas. Quomodo facti sunt in desolationem! * Subito defecerunt, perierunt prae horrore. Velut somnium evigilantis, Domine, * surgens imaginem ipsorum contemnes. Quia exacerbatum est cor meum, * et renes mei compuncti sunt; et ego insipiens factus sum et nescivi: * ut iumentum factus sum apud te. Ego autem semper tecum; * tenuisti manum dexteram meam. In consilio tuo deduces me * et postea cum gloria suscipies me. Quis enim mihi est in caelo? * Et tecum nihil volui super terram. Defecit caro mea et cor meum; * Deus cordis mei, et pars mea Deus in aeternum. [293] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 419 Quia ecce, qui elongant se a te, peribunt; * perdidisti omnes, qui fornicantur abs te. Mihi autem adhaerere Deo bonum est, * ponere in Domino Deo spem meam, ut annuntiem omnes operationes tuas * in portis filiae Sion. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Nobis adhaerere Deo bonum est, ponere in Deo spem nostram. Oratio (Br Goth 349, 409, 678; T35.2, 367, 635; LOPs 84) Nobis adhaerere Deo bonum est, tibi Domino bonum est; sed tu desiderium boni sic in nobis accumula, ut spes, quae nos tibi associat, nulla fidei titubatione vacillet, sed caritatis stabilitate perduret. R/ Amen. Concedente clementia sua, qui in Trinitate, unus Deus, vivit et regnat in saecula saeculorum. R/ Amen. Psalmus 105 Antiphona (Br Goth 321; T35.2, 368) (Ps 105, 4) Memento nostri, Domine, in beneplacito tuo. Confitemini Domino, quoniam bonus, * quoniam in saeculum misericordia eius. Quis loquetur potentias Domini, * auditas faciet omnes laudes eius? Beati, qui custodiunt iudicium * et faciunt iustitiam in omni tempore. Memento nostri, Domine, in beneplacito populi tui, * visita nos in salutari tuo, ut videamus bona electorum tuorum, + ut laetemur in laetitia gentis tuae, * ut gloriemur cum hereditate tua. Peccavimus cum patribus nostris, * iniuste egimus, iniquitatem fecimus. Patres nostri in Aegypto non intellexerunt mirabilia tua, + non fuerunt memores multitudinis misericordiarum tuarum * et irritaverunt ascendentes in mare, mare Rubrum. Et salvavit eos propter nomen suum, * ut notam faceret potentiam suam. Et increpuit mare Rubrum, et exsiccatum est, * et deduxit eos in abyssis sicut in deserto. 420 GABRIEL RAMIS MIQUEL [294] Et salvavit eos de manu odientis * et redemit eos de manu inimici. Et operuit aqua tribulantes eos: * unus ex eis non remansit. Et crediderunt verbis eius * et cantaverunt laudem eius. Cito obliti sunt operum eius * et non sustinuerunt consilium eius; et concupierunt concupiscentiam in deserto * et tentaverunt Deum in inaquoso. Et dedit eis petitionem ipsorum * et misit saturitatem in animas eorum. Et zelati sunt Moysen in castris, * Aaron sanctum Domini. Aperta est terra et deglutivit Dathan * et operuit super congregationem Abiram. Et exarsit ignis in synagoga eorum, * flamma combussit peccatores. Et fecerunt vitulum in Horeb * et adoraverunt sculptile; et mutaverunt gloriam suam * in similitudinem tauri comedentis fenum. Obliti sunt Deum, qui salvavit eos, * qui fecit magnalia in Aegypto, mirabilia in terra Cham, * terribilia in mari Rubro. Et dixit quia disperderet eos, * nisi affuisset Moyses electus eius: stetit in confractione in conspectu eius, * ut averteret iram eius, ne destrueret eos. Et pro nihilo habuerunt terram desiderabilem, * non crediderunt verbo eius. Et murmuraverunt in tabernaculis suis, * non exaudierunt vocem Domini. Et elevavit manum suam super eos, * ut prosterneret eos in deserto et ut deiceret semen eorum in nationibus * et dispergeret eos in regionibus. Et adhaeserunt Baalphegor * et comederunt sacrificia mortuorum; et irritaverunt eum in adinventionibus suis, * et irrupit in eos ruina. Et stetit Phinees et fecit iudicium, * et cessavit quassatio, et reputatum est ei in iustitiam * in generationem et generationem usque in sempiternum. Et irritaverunt eum ad aquas Meriba, * et vexatus est Moyses propter eos, [295] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 421 quia exacerbaverunt spiritum eius, * et temere locutus est in labiis suis. Non disperdiderunt gentes, * quas dixit Dominus illis. Et commixti sunt inter gentes * et didicerunt opera eorum. Et servierunt sculptilibus eorum, * et factum est illis in scandalum. Et immolaverunt filios suos * et filias suas daemoniis. Et effuderunt sanguinem innocentem, + sanguinem filiorum suorum et filiarum suarum, * quas sacrificaverunt sculptilibus Chanaan. Et infecta est terra in sanguinibus, + et contaminati sunt in operibus suis * et fornicati sunt in adinventionibus suis. Et exarsit ira Dominus in populum suum * et abominatus est hereditatem suam et tradidit eos in manus gentium, * et dominati sunt eorum, qui oderunt eos. Et tribulaverunt eos inimici eorum, * et humiliati sunt sub manibus eorum. Saepe liberavit eos; + ipsi autem exacerbaverunt eum in consilio suo * et corruerunt in iniquitatibus suis. Et vidit tribulationem eorum, * cum audivit clamorem eorum. Et memor fuit testamenti sui * et paenituit eum secundum multitudinem misericordiae suae. Et dedit eos in miserationes * in conspectu omnium, qui captivos duxerant eos. Salvos nos fac, Domine Deus noster, * et congrega nos de nationibus, ut confiteamur nomini sancto tuo * et gloriemur in laude tua. Benedictus Dominus, Deus Israel, + a saeculo et usque in saeculum. * Et dicet omnis populus: “Fiat, fiat”. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Memento nostri, Domine, in beneplacito tuo. Oratio (Br Goth 321, 428; T35.2, 369, 139; LOPs 121) Memento nostri, Domine, in beneplacito populi tui, et in tuo nos visita salutari; ut qui nostris cum patribus coram te inique gessimus, a damnationis origine et actualis peccati contagione, tua reduce gratia liberemur. R/ Amen. 422 GABRIEL RAMIS MIQUEL [296] Concedente gratia miserationis illius, cuius regnum et imperium iuge ac gloriosum in saecula saeculorum. R/ Amen. Responsorium (Br Goth 349; T35.2, 370; AL 181) (Cfr. Ps 125, 1; Lam 5, 16; cfr. Ps 73, 7; Ps 78, 1; Ez 22, 26; 23, 38; Is 64, 1a; cfr. Is 64, 2b; 63, 19) Facti sumus, Domine, tamquam alieni a te; vae nobis quia peccavimus, inimici tui polluerunt sancta tua. Utinam disrumperes caelos, et descenderes; et innotesceres nomen tuum, et in ira tua gentes conturbarentur. V/ Facti sumus quasi in principio, cum non dominaneris nostri, neque invocaretur nomen tuum super nos. R/ Utinam disrumperes caelos, et descenderes; et innotesceres nomen tuum, et in ira tua gentes conturbarentur. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Utinam disrumperes caelos, et descenderes; et innotesceres nomen tuum, et in ira tua gentes conturbarentur. Psalmus L Antiphona (Br Goth 349; T35.2, 371; AL 185) (Cfr. Ps 50, 3.4.11) Secundum multitudinem miserationum tuarum lava me ab iniustitia mea; mala mea dimitte, et omnes iniquitates meas dele. (Ut in ordinario in officio feriali) Ant. Secundum multitudinem miserationum tuarum lava me ab iniustitia mea; mala mea dimitte, et omnes iniquitates meas dele. Oratio (Br Goth 349; T35.2, 372; MPs 22, 313; LOPs 365) Miserator Domine, secundum magnam misericordiam tuam, nobis propitius miserere; ut quorum te non latent occulta, his de te confidentibus donetur misericordia manifesta; [297] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 423 quique pro admissis interno rugitu gemimus, et pro remissione peccatorum nostrorum criminum suspiramus, non nos a facie tua proicias, sed tibi soli debita pietate ad indulgentiae portum adducas. R/ Amen. Tua protegente misericordia, Deus noster, qui vivis et regnas in saecula saeculorum. R/ Amen. Canticum VT (Br Goth 349-350) (Iob 3, 3-26) Antiphona (Br Goth 349; T35.2, 373) (Cfr. Iob 3, 24) Quasi inundantes aquae, sic rugitus noster ante te, Domine Deus noster. “Pereat dies, in qua natus sum, * et nox, in qua dictum est: “Conceptus est homo”. Dies ille vertatur in tenebras; + non requirat eum Deus desuper * et non illustretur lumine. Obscurent eum tenebrae et umbra mortis; + occupet eum caligo * et involvatur amaritudine. Noctem illam tenebrosus turbo possideat; + non computetur in diebus anni, * nec numeretur in mensibus. Sit nox illa solitaria nec laude digna; + maledicant ei, qui maledicunt diei, * qui parati sunt suscitare Leviathan. Obtenebrentur stellae crepusculi eius; + expectet lucem et non sit, * nec videat palpebras aurorae, quia non conclusit ostia ventris, qui portavit me, * nec abstulit mala ab oculis meis. Quare non in vulva mortuus sum? * Egressus ex utero non statim peri? Quare exceptus genibus? * Cur lactatus uberibus? Nunc enim dormiens silerem * et somno meo requiescerem cum regibus et consulibus terrae, * qui aedificant sibi solitudines; aut cum principibus, qui possident aurum * et replent domos suas argento. Aut sicut abortivum absconditum non subsisterem, * vel qui concepti non viderunt lumen. Ibi impii cessaverunt a tumultu, * et ibi requieverunt fessi robore. 424 GABRIEL RAMIS MIQUEL [298] Et quondam vincti pariter sine molestia * non audierunt vocem exactoris. Parvus et magnus ibi sunt, * et servus liber a domino suo. Quare misero data est lux * et vita his, qui in amaritudine animae sunt? Qui expectant mortem, et non venit, * et effodiunt quaerentes illam magis quam tesauros; gaudentque vehementer * et laetantur sepulcro. Viro, cuius abscondita est via, * et circumdedit eum Deus tenebris. Antequam comedam suspiro, * et quasi inundantes aquae sic rugitus meus. Quia timor, quem timebam, evenit mihi, * et, quod verebar, accidit. Non dissimulavi, non silui, non quievi, * et venit super me indignatio”. Gloria et honor Patri et Filio et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Quasi inundantes aquae, sic rugitus noster ante te, Domine Deus noster. Oratio de Cantico (Br Goth 350) Quamdiu sumus, Domine, in huius vitae exilio, nec aut malitiae nequitia vigeat, nec nostrae virtutis robur enervet, aut precipitatis irae turbulenta commotio tranquillitatem nostrae mentis obscuret, aut candorem munditiae carnalium voluptatum delectatio coenosa commaculet; sed futuri iudicii praestolatio ab omnibus inquinamentis ac sordibus nos emundet. R/ Amen. Per misericordiam ipsius Dei nostri, qui vivit et omnia regit in saecula saeculorum. R/ Amen. Psalmus matutinarius (Br Goth 350; T35.2, 374) (Cfr. Ps 66, 3.2) Illumina, Domine, vultum tuum super nos, et miserere nobis. Ut cognoscamus in terra viam tuam. V/ Deus misereatur nobis, et benedicat nos; illuminet Dominus vultum suum super nos. R/ Ut cognoscamus in terra viam tuam. [299] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 425 V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Ut cognoscamus in terra viam tuam. Psalmi laudate 148-150 Antiphona (Br Goth 350; T35.2, 375; cfr. AL 181) (Cfr. Ps 148, 5a.9) Laudate nomen Domini montes et omnes colles, ligna fructífera et omnes cedri. (Ut in ordinario in officio feriali) Ant. Laudate nomen Domini montes et omnes colles, ligna fructífera et omnes cedri. Lectio (Br Goth 350-351) (Gn 49, 28-50) Lectio libri Genesis. R/ Deo gratias. In illis diebus: Benedixit Iacob filiis suis benedictionibus propriis […] repositus est in loculo in Aegypto. R/ Amen. Hymnus (Br Goth 201; T35.2, 111) Deus Pater ingenite et Fili unigenite, quos Trinitas unitas Sancto connectit Spiritui. Nos lucis ortus admonet, grates deferre debitas; tibique laudes dicere, quod nox obscura praeterit. Te frustra nullus invocat, nec cassis umquam vocibus amator tui luminis ad caelum vultus erigit. Diem precamur bonum; ut nostros, Salvator, actus sinceritati perpeti pius benigne instruas. Et tu suspirantem, Deus, vel vota supplicantium, vel corda confitentium, semper benignus aspice. Deo Patri sit gloria, eiusque soli Filio, cum Spiritu Paraclito in sempiterna saecula. Amen. Versus (AL 158, 170) (Ps 62, 7-8) V/ In matutinis meditabor in te. R/ Quia factus es adiutor meus, Domine. 426 GABRIEL RAMIS MIQUEL [300] Supplicatio (Br Goth 259) Oremus. Redemptorem mundi, Dominum nostrum Iesum Christum, cum omni supplicatione rogemus; ut nos, gratia ipsius, et per intercessionem sanctae Mariae Virginis, et Genetricis suae, ut sancto tempore quadragesimae, nobis remissionem peccatorum, et pacem nobis donare dignetur. R/ Praesta aeterne omnipotens Deus. Kyrie eleison. R/ Christe eleison, Kyrie eleison. Completuria (Br Goth 87; T35.2, 378, 113, 201; LOPs 251) Illumina, Domine, vultum tuum super nos, et benedictionem tuam ubertim nostris infunde pectoribus; quo corda nostra, lucis tuae hilaritate perfusa, tuam in terris valeant agnoscere viam, et omnibus singulari gentibus fidei sacrae laetentur notitiam. R/ Amen. Per misericordiam tuam, Deus noster, qui es benedictus, et vivis et omnia regis in saecula saeculorum. R/ Amen. Pater noster (Ut in ordinario in officio feriali) Petitio (Ut in ordinario in officio feriali) Benedictio (Br Goth 341; T35.2, 314, 183) Diac. Humiliate vos bendictioni. Praesb. Dominus sit semper vobiscum. R/ Et cum spiritu tuo. Omnipotentis Domini vos gratia protegat, famemque vestram spirituali cibo reficiat. R/ Amen. Sitim quoque vestram iustitiae fluminibus irriget, atque novum dulcedinis suae animis nostris infundat saporem. [301] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 427 R/ Amen. Ut tam carne contriti quam spiritu vivificati, ab eo mereamini caelesti dono beatificari. R/ Amen. Per misericordiam ipsius Dei nostri, qui est benedictus, et vivit et omnia regit in saecula saeculorum. R/ Amen. Psallendum (Br Goth 352; T35.2, 379) (Ps 118, 122-123) Excipe servum tuum in bono, ut non calumnientur mihi superbi. V/ Oculi mei defecerunt in salutare tuum, et in eloquium iustitiae tuae. R/ Ut non calumnientur mihi superbi. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Ut non calumnientur mihi superbi. Oratio (Br Goth 352, 506, 539, 552; T35.2, 1522, 1496; LOPs 289) Iudicium et iustitiam doce nos facere, Domine, qualiter valeamus superborum calumnias conclucare; excipe servos tuos in bonum, qui sperant in salutare tuum; quorum interiores oculi Christi tui expectant regnum, hos partem habere facito in consortio electorum. R/ Amen. Per misericordiam tuam, Deus noster, qui es benedictus, et vivis et omnia regis in saecula saeculorum. R/ Amen. Conclusio (Ut in ordinario in officio feriali) Ad tertiam Lectio (Br Goth 352) (Eccli 10, 2-10; 11,1) Lectio libri Ecclesiastici Salomonis. R/ Deo gratias. Fili: Secundum iudicem populi […] et in medio magnatorum consedere illum faciet. R/ Amen. 428 GABRIEL RAMIS MIQUEL [302] Lectio (Br Goth 352-353) (Ex 1 – 2, 10) Principium libri Exodi. R/ Deo gratias. Haec sunt nomina filiorum Israel […] vocavitque nomen eius Moysen dicens: “Quia de aqua tuli eum”. R/ Amen. Psalmus 124 Antiphona (Br Goth 353; T35.2, 380) (Ps 124, 1) Qui confidunt in Domino, sicut mons Sion; non commovebitur in aeternum. Qui confidunt in Domino, sicut mons Sion: * non commovebitur, in aeternum manet. Ierusalem, montes in circuitu eius, + et Dominus in circuitu populi sui * ex hoc nunc et usque in saeculum. Quia non requiescet virga iniquitatis super sortem iustorum, * ut non extendant iusti ad iniquitatem manus suas. Benefac, Domine, bonis * et rectis corde. Declinantes autem per vias pravas + adducet Dominus cum operantibus iniquitatem. * Pax super Israel! Gloria et honor Patri, et Filio, et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Qui confidunt in Domino, sicut mons Sion; non commovebitur in aeternum. Oratio (Br Goth 353, 158, 339; T35.2, 381, 300; LOPs 188) Benefac, Domine, bonis, et rectis corde regimen salutaris impende; atque in tuae plebis custodia semper vigila, et malignantium rabiem tua dextera frena. R/ Amen. Adiuvante sua misericordia, qui vivit et regnat, Deus, in saecula saeculorum. R/ Amen. Psalmus 81 Antiphona (Br Goth 353; T35.2, 382; cfr. AL 169) (Cfr. Ps 81, 8) Surge Deus, iudica terram, qui dominaris in omnibus gentibus. [303] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 429 Deus stetit in concilio divino, * in medio deorum iudicat. “Usquequo iudicabitis inique * et facies peccatorum sumetis? Iudicate egeno et pupillo, * humilem et pauperem iustificate. Eripite pauperem * et egenum de manu peccatoris liberate”. Nescierunt neque intellexerunt, in tenebris ambulant; * movebuntur omnia fundamenta terrae. Ego dixi: “Dii estis, * et filii Excelsi omnes”. Vos autem sicut homines moriemini * et sicut unus de principibus cadetis. Surge, Deus, iudica terram, * quoniam tu hereditabis in omnibus gentibus. Gloria et honor Patri, et Filio, et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Surge Deus, iudica terram, qui dominaris in omnibus gentibus. Oratio (Br Goth 353-354, 381, 417; T35.2, 383, 680; T35.5, 184; OV 531; LOPs 396) Exurge, Domine, qui iudicas terram; et qui fidem omnium gentium pius possessor inhabitas, ne patiaris nos in tenebris remanere; sed fac nos lumen tuae veritatis aspicere; ut fundamentum fidei nostrae, non super arenam, quam turbo deiiciat, aedificemus, sed super petram, cuius tu es firmitas collocemus. R/ Amen. Adiuvante sua misericordia, qui in Trinitate, unus Deus, gloriatur in saecula saeculorum. R/ Amen. Psalmus 82 Antiphona (Br Goth 354; T35.2, 384; cfr. AL 237) (Cfr. Ps 82, 2) Deus, quis similis tibi? ne taceas neque compescaris Deus. Deus, ne quiescas, ne taceas * neque compescaris, Deus, quoniam ecce inimici tui fremuerunt, * et, qui oderunt te, extulerunt caput. Adversus populum tuum malignaverunt consilium * et cogitaverunt adversus eos, quos abscondisti tibi. Dixerunt: “Venite, et disperdamus eos de gente, * et non memoretur nomen Israel ultra!”. 430 GABRIEL RAMIS MIQUEL [304] Quoniam cogitaverunt unanimiter, * adversum te testamentum statuerunt: tabernacula Idumaeorum et Ismaelitae, * Moab et Agareni, Gebal et Ammon et Amalec, * Philistaea cum habitantibus Tyrum. Etenim Assur sociabatur cum illis; * facti sunt in adiutorium filiis Lot. Fac illis sicut Madian et Sisarae, * sicut Iabin in torrente Cison. Disperierunt in Endor, * facti sunt ut stercus super terram. Pone duces eorum sicut Oreb et Zeb * et Zebee et Salmana, omnes principes eorum, qui dixerunt: * “Hereditate possideamus pascua Dei!”. Deus meus, pone illos ut rotam * et sicut stipulam ante ventum. Sicut ignis, qui comburit silvam, * et sicut flamma devorans montes, ita persequeris illos in tempestate tua * et in procella tua turbabis eos. Imple facies eorum ignominia, * et quaerent nomen tuum, Domine. Erubescant et conturbentur in saeculum saeculi * et confundantur et pereant; et cognoscant quia nomen tibi Dominus: * tu solus Altissimus super omnem terram. Gloria et honor Patri, et Filio, et Spiritui Sancto, * in saecula saeculorum. Amen. Ant. Deus, quis similis tibi? ne taceas neque compescaris Deus. Responsorium (Br Goth 354; T35.2, 385) (Cfr. Ier 8, 6.5) Attendite et auscultabitis, dicit Dominus; nemo quod bonum est loquitur. Nullus est qui agat paenitentiam super peccato suo. V/ Quare aversus est populus iste in Ierusalem, aversione contentiosa? Apprehenderunt mendacium, et noluerunt reverti. R/ Nullus est qui agat paenitentiam super peccato suo. V/ Gloria et honor Patri et Filio et Spiritui Sancto, in saecula saeculorum. Amen. R/ Nullus est qui agat paenitentiam super peccato suo. [305] RECONSTRUCCIÓN DEL BREVIARIO HISPANO-MOZÁRABE … 431 Preces (Br Goth 354) Diac. Flectamus genua. Presb. Deprecemur Dominum. R/ Kyrie eleison. Christe eleison. Kyrie eleison. Diac. Erigite vos. Tu, misericors Domine, eripe nos a flagello irae tuae, propitiare et miserere nobis.	26,140
tel-03771553-2022LIMO0054.txt_7	French-Science-Pile	Open Science	Various open science	2,022	Etude expérimentale et rationnelle de la dégradation des couches de surfaces de chaussées. Génie civil. Université de Limoges, 2022. Français. ⟨NNT : 2022LIMO0054⟩. ⟨tel-03771553⟩	None	French	Spoken	7,559	14,439	ΔA (mm2) ΔB (mm2) SF BBSG 287129,5 81836,6 3,50 BBAO 41153,5 37101 1,10 BBAO+ 69194 68000 1,02 Tableau IV-1 : Paramètres de caractérisation des courbes d’orniérage IV.2.3.1. Influence de la température La Figure IV-9a illustre l’évolution de l’ornière après 100.000 cycles pour les couches de BBAO et de BBAO+. Pour le BBAO, l’augmentation de la température influence fortement la stabilité du mélange à haute température. La profondeur d’ornière a significativement augmenté lorsque la température passe de 30 à 35°C. Durant cette phase d’essai, le taux d’évolution de l’ornière passe de 1 mm à 1. 5 mm. Par ailleurs, quand les essais ont été réalisés entre 45 et 55°C, le taux d’augmentation de ce dernier est de 2,5 mm et l’augmentation de la température de 30°C à 55°C conduit à une augmentation de l’orniérage de 40%. Dans cette phase, la température accélère le mouvement des granulats, qui se manifeste par des soulèvements latéraux, indiquant une rupture par cisaillement du mélange (Figure IV-9b). 159 Figure IV-9 : a) Courbes d’orniérage après 100.000 cycles, b) Ornière BBAO à 160000 cycles L’influence de la température sur la couche de BBAO+ est marquée par deux parties. La première partie concerne les essais réalisés à des températures plus ou moins élevés (entre 20°C et 30°C) alors que la deuxième partie concerne les basses températures (entre 12°C et 17°C). L’influence de la température sur le développement de l’orniérage dans la première partie est similaire aux phénomènes remarquées et expliquées pour le BBAO. Par ailleurs, à basses températures, le taux d’augmentation de ’orniérage est nul malgré l’augmentation du nombre de passage (Figure IV-9a). IV.3. Suivi de l’orniérage sur l’enrobé à froid (Agreco) L’objectif de ce paragraphe est d’étudier la performance de l’enrobé à froid vis–à–vis de l’orniérage. La Figure IV-10 illustre l’évolution de la profondeur de l’ornière en fonction du nombre de cycles de chargement à des températures comprises entre 13 et 30°C. Les températures relevées correspondent aux températures moyennes lors des essais. 160 Figure IV-10 : Courbe de l’orniérage de l’Agreco Les analyses montrent trois phases de développement de l’ornière. Dans la première phase, l’ornière augmente rapidement mais la vitesse d’orniérage tend à diminuer atteignant une valeur constante ou avec cinétique d’évolution très faible dans la deuxième phase (ce qui tend à considérer dans la profession les matériaux à froid anti-orniérants). Au cours de cette deuxième phase, on remarque une diminution de la vitesse d’évolution de l’ornière, qui est probablement due au murissement des enrobés à froid. La présence de ce phénomène réside dans le fait qu'il existe de nombreux défauts dans l’enrobé, à savoir le pourcentage assez important des vides qui de 15% dans ce mélange ainsi que des dislocations dans les agrégats. C'est le mouvement de ces dislocations, en particulier dans le liant bitumineux, sous chargement répété qui provoque les déformations permanentes des enrobés bitumineux. La densité des dislocations augmente progressivement avec des chargements répétés. Cependant, la fraction de ces dislocations mobiles diminue simultanément car les dislocations croissantes se rencontrent fréquemment. Par conséquent, les intersections de dislocations peuvent réduire leur mobilité, ce qui entraîne une réduction progressive de la déformation permanente par cycle. Ainsi, le processus physique de l'étape primaire est le mouvement (ou microflux) des dislocations, généralement appelé écrouissage. Avec l'accumulation des déformations permanentes, l’Agreco durcit sous des charges répétées jusqu'à ce qu'il atteigne un point où les microfissures s'amorcent et se développent. La déformation permanente de chaque cycle cesse alors de diminuer et le stade secondaire ou stable apparaît. L'apparition de microfissures entraîne le développement de plus de dislocations. Dans le même temps, ce processus fournit également « l’espace » supplémentaire pour la mobilité des dislocations et ramollit davantage l’enrobé bitumineux. L'étape secondaire est atteinte lorsque la vitesse d'écrouissage est dynamiquement égale à celle de l'adoucissement. L'équilibre dynamique se traduit par l'accumulation linéaire des déformations permanentes au cours de l'étape 161 secondaire. Enfin, la vitesse d’orniérage augmente brutalement avec l’augmentation de la température qui passe de 15°C à 30°C, il apparait clairement une cinétique concave de rupture. IV.1. Modèle prédictif de l'orniérage basé sur les essais à grande échelle Lorsque les conditions environnementales ne changent pas de manière appréciable, la plupart des études d'essais de chaussées empiriques et accélérées présentent une tendance concave de déformation (profondeur d'ornière) par rapport au nombre d'applications de charge. La courbe A de la Figure IV-11 montre cette tendance. Elle illustre la façon dont les matériaux durcissent avec le chargement. Le même incrément du nombre d'applications de charge (DN) produit un incrément plus important de profondeur d'ornière (DRD) vers le début que vers la fin de la durée de vie de la chaussée (Archilla & Madanat, 2001). La courbe B illustre ce qui se passe lorsque l'environnement passe à des conditions plus défavorables. Par exemple, le phénomène de dégel affecte négativement la performance des matériaux non liés, et les températures élevées de l'air affectent négativement la performance des mélanges des enrobés bitumineux. Dans de telles situations, la tendance de la courbe A est interrompue par les sauts dans les incréments de profondeurs d'ornière indiqués par la courbe B. De cette manière, les sauts de profondeur d'ornière avec des changements des conditions environnementales peuvent être pris en compte dans la spécification des incréments de profondeur d'ornière. Pour modéliser les incréments de profondeur d'ornière pour chaque période, il est important de considérer les mécanismes d'orniérage et les emplacements d'origine des déformations dans la structure de la chaussée. L'orniérage apparaît comme une conséquence de la densification et de l'écoulement plastique dans les couches de chaussée. Il est très difficile d'attribuer la part d'orniérage due à la densification et la part due à l'écoulement car les deux mécanismes se produisent dans toutes les couches de chaussée à des degrés différents. Cependant, près de la surface de chaussée, l'écoulement joue un rôle plus important que la densification, mais l'importance de l'écoulement par rapport à la densification diminue avec la profondeur. Ainsi, bien que les deux mécanismes ne puissent être séparés, il faut s'attendre à des compromis différents entre les variables décrivant ces phénomènes près de la surface de la chaussée et celles en profondeur de la structure de chaussée. 162 Figure IV-11 : Profondeur de l’orniérage en fonction du nombre de cycle (d'après Archilla & Madanat, 2001) Par conséquent, l'incrément de profondeur d'ornière pour chaque période est modél isé comme la somme incrément pre naissance proche de environnement sont différents. Proche de la surface, l’enrobé bitumineux a des performances fortement affectées par des températures élevées de l'air. En revanche, les couches sous-jacentes de la structure de chaussée sont généralement construites avec des matériaux non liés qui sont plutôt affectés par les phases de gel - dégel. IV.1.1. Modélisation des performances d'orniérage Pour caractériser la sensibilité de l’ornière vis-à-vis du nombre de cycles de chargement et de la température, nous avons effectué une étude statistique sur les mesures d’ornière pour les quatre matériaux. L’analyse statistique est basée sur une étude de la régression linéaire des variables suivants : Rd (profondeur de l’ornière), T (température) et N (nombre de cycles). Les résultats de l’analyse des corrélations de Pearson (valeur P) sont présentés dans le Tableau IV-2. On remarque que la profondeur de l’ornière (Rd) a une corrélation plus élevée avec la température (T) qu'avec le nombre de cycle (N). En utilisant les résultats des relevés de la profondeur de l’ornière, nous avons effectué une analyse de régression multiple pour un total de 10 valeurs expérimentales en utilisant les trois conditions de température, le nombre de cycles et la profondeur d’ornière comme paramètres. Sur la base de notre analyse, nous avons généré un modèle de prédiction de l'orniérage exprimé dans l'équation IV-3. Le Tableau IV-3 répertorie les valeurs prédites des paramètres générés par l'analyse de la régression multiple ainsi que le coefficient de corrélation et la proportion significative (valeur P). 163 BBSG N T Rd BBAO N T Rd BBAO+ N T Rd Agreco N T Rd (résultats exprimés en %) N T Rd Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation -0.8 0.04 6 0.5 0.0005 6 -0.8 0.04 6 0.98 0.0005 6 0.5 0.0005 6 0.98 0.0005 6 - (résultats exprimés en %) N T Rd Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation -0.84 0.01 6 0.38 0.009 6 -0.84 0.01 6 0.78 0.007 6 0.38 0.009 6 0.78 0.007 6 - (résultats exprimés en %) N T Rd Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation -0.87 0.001 6 0.3 0.009 6 -0.87 0.001 6 0.58 0.005 6 0.3 0.009 6 0.58 0.005 6 - (résultats exprimés en %) N T Rd Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation Coefficient de corrélation Valeur P Nb de point d’observation -0.70 0.02 6 0.28 0.0001 6 -0.70 0.02 6 0.78 0.002 6 0.28 0.0001 6 0.78 0.002 6 Tableau IV-2 : Corrélations entre divers paramètres (N,T,Rd) Rd =αT β Nδ Eq IV-3 Avec Rd la profondeur de l'ornière, T la température et N le nombre de cycles. Les paramètres α, β et δ sont des paramètres propres à chaque matériau. α dépend de la rigidité du matériau (plus le module de rigidité est faible plus α est important). β indique la sensibilité du matériau à la température et δ montre sa sensibilité au nombre de cycles. BBSG α=0.0035 β=0.87 δ=0.43 Statistique t 1.95 9.85 4.064 p-valeur BBAO α=0.0004 β=1.01 δ=0.5 Statistique t 2.43 4.13 2.6 p-valeur BBAO+ α=0.0001 β=1.01 δ=0.5 Statistique t 6.4 2.13 0.9 p-valeur Agreco α=0.031 β=0.62 δ=0.43 Statistique t 3.41 4.13 2.71 p-valeur <0.001 <0.001 <0.001 <0.001 Erreur-type -0.48 0.00001 0.07 Erreur-type -0.02 1.25 0.00007 Erreur-type -0.4 2.68 3.71 Erreur-type -1 1.25 1.71 Tableau IV-3 : Résultats de l'analyse de régression multiple sur les données La Figure IV-12 montre une comparaison des valeurs prédites et les mesures. Les comparaisons montrent que les modèles proposés pour prédire la profondeur de l'orniérage ont montré une bonne corrélation avec les résultats expérimentaux. Les modèles proposés sont valables pour les températures croissantes. Par ailleurs, l’analyse des équations montre une évolution quasi-linéaire de l’ornière en fonction de la température et une évolution « racine-carré » en fonction du nombre de cycle de chargement. Figure IV-12: Comparaison entre l’orniérage calculé et mesuré pour les quatre matériaux IV.1.2. Discussion des modèles et analyse des paramètres Pour déterminer la sensibilité des paramètres dans le modèle proposé, nous avons réalisé une analyse paramétrique afin d’estimer leur impact sur l’orniérage. IV.1.2.1. Effet du paramètre α La Figure IV-13 montre l’effet du coefficient sur l’évolution de la profondeur de l’orniérage en fonction du nombre de cycles. L’augmentation de α implique une augmentation rapide de l’orniérage. Pour cette simulation, α traduit la vitesse du développement de l’orniérage pour une température constante (T=20°C et β=1) avec augmentation du nombre de cycle. Figure IV-13 : Étude paramétrique sur α 166 Le paramètre β caractérise la sensibilité du matériau à la température. La Figure IV-14 montre l’influence de la variation de β sur l’évolution de la profondeur de l’ornière. Pour un coefficient α =0,0005 et δ =0.87, le paramètre β engendre un effet significatif sur l’orniérage. Pour chacune des valeurs, l’augmentation de ce dernier provoque une augmentation drastique de la profondeur de l’ornière. Ce paramètre engendre aussi un changement de la courbe d’ornière, plus le coefficient est important plus la tangente à la courbe est importante. Figure IV-14 : Étude paramétrique sur β IV.1.2.2. Effet du paramètre δ Le paramètre δ caractérise l’effet du nombre de cycles sur la profondeur de l’ornière. La Figure IV-15 montre l’évolution de l’ornière en fonction de la variation de δ. Son augmentation n’a un effet significatif qu’à partir δ = 0.6. La vitesse d’évolution de la profondeur n’est pas influencée par l’évolution de δ jusqu’à cette même valeur. Figure IV-15 : Étude paramétrique du paramètre δ IV.1.3. Principe d’équivalence temps - température appliqué à l’orniérage L’objectif de ce paragraphe est d’appliquer le Principe d’Equivalence Temps-Température (Di Benedetto et al., 2008) à partir des modèles de prédiction de l’orniérage établie dans le paragraphe précédent. D ‘autre part, Khosravifar et al. (Khosravifar et al., 2015) ont étudié la 167 possibilité d'utiliser la fonction de translation de température obtenue à partir des essais de module dynamique pour construire une courbe maîtresse des déformations et pour prédire la déformation cumulée en termes de cycles de chargement réduits. La figure IV-16 B illustre la déformation axiale plastique par rapport au nombre réduit de cycles de charge (NR) dans l'espace log – log. Les déformations axiales plastiques à différentes températures se translatent relativement bien pour former une seule déformation par rapport à un nombre réduit de cycles de chargement, confirmant la validité de PETT dans le domaine viscoplastique dans essais uniaxial sous chargements répétés. Les lignes en pointillés de la figure IV-16B présentent la prédiction de deuxième phase de la courbe de l’orniérage basée sur des essais à différentes températures mais après les avoir déplacées à la température de référence de 20°C. Figure IV-16 : PETT appliquée au essais uniaxial sous chargements répétés à trois températures différentes (à gauche), Courbe maîtresse des deformations permanente à Tref = 20°C (à droite). (D’après Khosravifar et al ., 2015) Là où le PETT s’applique, à partir des modèles issus des essais en vraies grandeurs à une température et à un nombre de cycles donnés (Ti,NTi), l’évolution de la profondeur de l’ornière reste la même pour un essai effectué à un nombre de cycles et une température de référence (Tref,NTref). Le nombre de cycles équivalent est exprimé par le coefficient de translation décrit comme suit : NTref≡Nref = NTi ∗ aT Eq IV-4 La Figure IV- schématise le PETT appliqué au modèle issu des essais en vraies grandeurs : 168 Figure IV-17 : PETT appliqué au modèle d’orniérage Pour établir le PETT, nous allons utiliser le modèle de prédiction de l’orniérage du BBAO définit comme suit : RD(BBAO) = 0,0004 ∗ N 0,50 ∗ T 1,01 Eq IV-5 D’après l’équation Eq IV-5 et pour la même profondeur de l’ornière la relation devient : Nref log(0,0004 ∗ Nref 0,50 ∗ Tref 1,01 ) = log(0,0004 ∗ ( aT )0,50 ∗ Ti 1,01 ) Eq IV-6 Il est possible de ramener l’équation IV-6 à une forme plus simple : Ti ) Tref Log(aT) = 2 × Log ( Eq IV-7 La fonction de translation aT peut se définir à partir de la relation entre la température et le Logarithme de aT. La Figure IV- montre l’évolution du coefficient de translation en fonction de la température pour une température choisie à 25°C. A l’échelle semi-logarithmique la fonction du coefficient aT est linéaire avec une pente positive. Figure IV-18 : Relation entre la température et le logarith me du facteur aT à partir de l’équation IV-7 Dans la littérature il existe plusieurs expressions pour exprimer l’évolution du coefficient de translation en fonction de la température. Cependant dans le domaine des enrobés bitumineux c’est le modelé WLF qui est largement utilisé car il traduit correctement le comportement des enrobé bitumineux. Pour ce faire nous essayé de déterminer les paramètres du modèle WLF défini dans l’équation VI-8 −C (T−T ) 1 R log(aT ) = (T−T )+C R Equation VI-8 2 Pour ce faire on s’est basé sur le modèle développé dans l’équation VI-7 pour déterminer les paramètres du modèle C1 et C2. La méthode suivie consiste à l’optimisation de l’erreur entre les deux modèles (Equation IV-7et 8) qui a été fait sur le module Solveur sur Excel pour une température de référence de 20°C.A partir de cette analyse on détermine les coefficient C1 et C2 définit comme suit :C1= -2,1 ; C2= 50.Les travaux entreprirent par Andriescu et Hesp (Andriescu & Hesp, 2009) pour l’application du PETT aux déformations permanentes Ce qui semble cohérent avec nos résultats. La figure IV-19 montre la comparaison entre le modèle issu de notre calcul est le modèle WLF. Les résultats montrent une très bonne superpositions 170 entre les deux modèles et que la fonction log( aT ) est une fonction croissante en fonction de la température. Figure IV-19 : Comparaison entre le modèle analytique et le modèle WLF Comme expliqué dans la méthodologie de l’utilisation du PETT, la fonction de translation et d'ajustement de la courbe maîtresse est utilisée pour décaler horizontalement la profondeur de l’ornière à chaque température pour former la courbe maîtresse à la température de référence de 25°C. La Figure IV- représente l’ornière en fonction du nombre de cycles dans l'espace log-log. Figure IV-20 : Construction de la courbe maitresse La profondeur de l’orniérage à différentes températures diminue pour former une seule courbe par rapport à un nombre de cycles de chargement. Cela confirme la validité du PETT dans le domaine viscoplastique. Les droites présentent l’évolution de l’ornière prédites à partir du modèle à différentes températures après avoir été déplacées à la température de référence de 20°C. 171 Afin d’exploiter le PETT pour réduire le nombre d’essai et pour suivre l’évolution de l’ornière, il est important de prédire efficacement ce dernier à n'importe quelle température en utilisant les résultats des essais à partir d’une température constante. Pour valider cela, nous avons une température de référence de 20°C et nous essayons de tracer la courbe d’orniérage à une température constante de 30°C et une courbe d’orniérage à trois température différentes. La figure IV-21 montre la comparaison entre le palier de la courbe de l’orniérage BBAO à 30°C et l’orniérage calculé à partir du PETT pour la même température et le même de nombre de cycle. Les résultats montrent une bonne corrélation entre les calculs et les mesures. L’erreur de prédiction à partir du PETT est inférieur à 6%. Figure IV-21 : Comparaison entre mesure et calcul issu du PETT à une température constante (30°C) Le deuxième cas de la validation du PETT est d’essayer de tracer une courbe d’ornière à des paliers de température différentes. Nous avons choisi la dernière partie de la courbe d’orniérage du BBAO dont les essais ont été déroulés entre 30 et 40 °C. La figure IV-22 montre les résultats de la comparaison entre les mesures et les calculs issus du PETT. Figure IV-22 : Comparaison entre mesure et calcul issu du PETT à des températures différentes L’application du principe d’équivalence temps température permets de prédire d’une manière satisfaisante l’orniérage à différentes températures aussi, l’erreur entre les mesures et l’orniérage calculé étant inférieur à 3.5%. 172 D’autre part l’application du PETT à l’orniérage permets de prédire des courbes d’orniérage à des températures constantes (Figure IV-23) et de prévoir l’évolution de l’ornière pour n’importe quelle situation, cependant cette configuration doit faire objet à une validation expérimentale. Le contrôle de la température et le maintien d’une température étant possible dans notre halle à l’aide des panneaux chauffants pilotés en température. Figure IV-23 : Courbes d’orniérages calculées à différentes températures IV.2. Conclusion du chapitre IV Dans ce dernier chapitre, nous avons étudié les mécanismes d'orniérage de quatre matériaux de surface différents. Les profils d'orniérage des quatre matériaux de surface présentent une forme en W. Le BBAO+ a une performance d'orniérage plus constante. L'analyse de la zone de déformation a suggéré que la déformation permanente la plus importante s'est produite principalement dans l’enrobé à froid. La performance vis à vis de l’orniérage dépend néanmoins du revêtement bitumineux et de sa rigidité. Un modèle de prédiction d'orniérage intégrant les essais accélérés a été développé. Ce modèle intègre le nombre de cycles et la température. Les résultats ont montré que l’orniérage est plus sensible à la température. Finalement, nous avons étudié la possibilité d'utiliser le principe d’équivalence temps température pour prédire la profondeur de l’orniérage. Conclusion générale et perspectives De nouvelles dégradations en surface des chaussées bitumineuses, telles que la fissuration par le haut et l’orniérage à petit rayon, causées par le passage répété de véhicules lourds ont conduit à s'interroger sur le comportement de la couche de surface liée au contact pneuchaussée. Aussi, pour augmenter la durée de vie de ces ouvrages, les futures méthodes de dimensionnement devront mieux prendre en compte l’impact du contact réel sur l'état de contrainte et de déformation de la surface de la chaussée. Dans l’objectif de proposer une future méthode rationnelle de dimensionnement de la couche de surface, ce document a présenté l’analyse du comportement mécanique de la couche de surface à l’aide d’essais en vraies grandeurs et un modèle numérique -analytique. Dans un premier temps, quatre matériaux de surface ont été soumis au simulateur de trafic MLS10. Les mesures ont été comparés aux résultats du modèle semi-analytique SAM et du logiciel ViscoRoute. Les différentes analyses en surface montrent alors que les déformations dépendent fortement de la position de la charge. Nous avons démontré que : - Au bord extérieur du pneumatique et dans la direction transversale, la couche de surface est sollicitée en traction. L’amplitude maximale des déformations en traction dépend principalement de la rigidité, de la température et de l’épaisseur de la structure. - Dans l’axe du jumelage, la couche de surface est sollicitée en compression. Le signal mesuré et calculé a été marqué par un saut de déformation dans la zone centrale provoqué par la contribution des contraintes dans la déformation transversale par effet Poisson. Plus le coefficient de Poisson est important plus le saut de déformation est marqué. Ce phénomène explique qu’en surface et dans l’axe du jumelage, l’enrobé bitumineux subit des contraintes longitudinales nettement supérieure aux contraintes transversales, ce qui signifie que le revêtement bitumineux est plus sollicité horizontalement que transversalement. - A l’interface de la couche de surface et la couche de grave bitume, les quatre matériaux ont été sollicités dans l’axe du jumelage en contraction et au bord extérieur du pneu en extension. L’amplitude maximale de la déformation en contraction dépend de la rigidité de l’enrobé bitumineux. A faible profondeur de la surface, un saut de déformation a été observé dans les signaux calculés et mesurés. - Les modélisations numériques réalisées sur SAM et ViscoRoute ont permis de prédire raisonnablement et de manière équivalente les déformations réelles en dehors de la zone de roulement (en dehors du contact). Par contre, dans l’axe du jumelage, les 174 modélisations élastiques et viscoélastique donnent des résultats moins satisfaisants. La complexité de l'état de déformation, la non-uniformité de la géométrie et de la structure du pneumatique et l'effet non linéaire des matériaux peuvent expliquer ces différences. - Les conditions d’interface ont un effet sur les modélisations, les interfaces viscoélastiques donnent une meilleure prédiction du comportement de la structure. Ensuite, les mécanismes de l’orniérage pour les quatre matériaux de surface ont été étudié et discuté comme suit : - L’analyse de la forme du profil d‘ornière a démontré que le béton bitumineux AntiOrniérant modifié + (BBAO+) présente une meilleure résistance au cisaillement que les BBAO et BBSG. L’analyse de l’évolution de la profondeur d’ornière en fonction du nombre de cycles a confirmé que le BBAO+ était beaucoup plus performant vis-à-vis de l’orniérage. L’étude de la zone de déformation a suggéré également que la déformation permanente la plus importante s'est produite principalement dans l’enrobé à froid Agreco. - Le suivi de l’ornière à l’aide des essais en vraies grandeurs à des températures contrôlées a contribué au développement de modèles empiriques de prédiction de l'orniérage. Ces modèles ont intégré le principe d’équivalence temps-températures pour une bonne concordance avec les résultats d’essais en vraies grandeurs. Ces modèles intègrent trois paramètres principaux : nombre de cycles, rigidité du matériau et érature. Par une analyse paramétrique, nous avons démontré que l’orniérage est plus sensible à la température. Enfin, le comportement de la couche de surface dans la zone de contact a été étudié pour le cas particulier d’un virage. Pour cela, les contraintes surfaciques ont été mesurées et simulées par des calculs Semi-Analytiques en intégrant la vraie géométrie du pneu. Nous avons montré que les pressions de contact dépendent fortement de l’angle d’inclinaison en virage. L’aire de contact pneu chaussée est devisé en deux zones : la zone d’adhérence à l'avant et la zone de glissement à l’arrière de l'aire de contact. Le comportement dans ces deux zones est différent. Les contraintes de cisaillement deviennent plus importantes avec l’augmentation de l’angle d’inclinaison jusqu’à produire une rupture par cisaillement dans les virages. La déformation principale est concentrée au niveau du bord intérieur du pneumatique. Elle augmente avec l'angle d'inclinaison. Ces déformations importantes démontrent alors que le virage est plus sensible aux fissurations descendantes. Bien que les travaux présentés dans ce document aient montré l’efficacité des modélisations et l’intérêt de l’utilisation d’un simulateur de trafic (MLS10) pour leur validation, nous devons 175 poursuivre par des analyses plus approfondies du comportement de la couche de surface en vue du développement d’un outil de calcul de dimensionnement. Une confrontation à des essais à petite échelle pourrait être entreprise. Une étude plus approfondie des caractéristiques en fatigue des matériaux s’appuyant sur les essais en laboratoire s’avère nécessaire. Elle permettrait de conforter les résultats de la durée de vie de la couche de surface vis-à-vis des fissurations descendantes et de l’orniérage. La poursuite du développement du code de calcul Semi-Analytique entreprit par Manyo (Manyo,2019), utilisant les résultats présentés dans cette thèse devra être fait. Cet outil numérique permettra la validation des essais en vraies grandeurs et permettra d’introduire une loi viscoplastique. 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W2786725705.txt_9	German-Science-Pile	Open Science	Various open science	null	None	None	Unknown	Spoken	7,668	8,668	In ferio r courts had fiv e ju d g es. s u p re m e c o u r t u n t i l 1 8 U5 . ) (G eorgia d id n ot have a C o u r t s e s s i o n s , w h ic h l a s t e d t w i c e a y e a r , t o o k p l a c e one a f t e r t h e o t h e r i n t h e ^ S t e p h e n s , R e c o l l e c t i o n s , 362-65; S t e p h e n s P a p e r s , LC. f i v e days county s e a t s AHS D i a r y , 22 J u l y 1 8 3 ^ , ^ J o h n s t o n and B r o w n e , S t e p h e n s , 9 0 ; C lev elan d , Stephens , 43. Two days a f t e r a d m i s s i o n t o t h e b a r , S t e p h e n s g e t h i s f i r s t e n g a g e m e n t as a l a w y e r — a t a c o n t i n g e n t f e e o f $ 1 8 0. C raw fordville, i t t u r n e d o u t , w o u l d p r o v i d e him w i t h a c o m f o r t a b l e l i v i n g. AHS D i a r y , 2k J u l y 1834, S t e p h e n s P a p e r s , LC. In h i s f i r s t y e a r o f p r a c t i c e S t e p h e n s made f o u r h u n d r e d d o l l a r s. C l e v e l a n d , S t e p h e n s , 44. 53 o f each c i r c u i t. Stephens p r a c t i c e d in the n o r th e r n c i r c u i t : co u n ties o f W ilk e s, Greene, E lb e r t, F r a n k lin , He was f o r t u n a t e t o b e i n t h i s th e most i l l u s t r i o u s and th e s t a t e ' s Colum bia, and O g le th o rp e. c i r c u i t b e c a u s e i t was f a r a n d away in the s t a t e , c o n ta in in g o u ts ta n d in g law yers sharpest p o litic ia n s : C h a r le s J e n k i n s , George C raw ford, a n d Andrew M i l l e r o f Ri c h m o n d , f o r e x a m p l e ; H o w e ll Cobb o f C l a r k e ; Ge or ge G i l m e r a nd J o s e p h Lumpkin o f O g l e t h o r p e ; J ames Thomas, F r a n c i s the G a r n e tt Andrews, Con e, W i l l i a m Dawson, R o b e r t Toombs, a n d o t h e r s. The n a t u r e o f t h e work t h r e w t h e s e members o f t h e l e g a l p r o f e s s i o n i n t o c lo s e c o n ta c t w ith each o t h e r. enm ities, formed in t h e v i l l a g e t a v e r n s la ste d for years. Stephens court, F r i e n d s h i p s , n o t to mention a trip and h o t e l s on t h e c i r c u i t o f t e n 27 f o r m e d s u c h a f r i e n d s h i p on h i s first trip to a c ir c u it re m a rk a b le f o r n o t h i n g e t h e r t h a n t h e f a c t t h a t he rode on a b o r r o w e d h o r s e and h a d t o c h a n g e i n t o h i s o n l y p r e s e n t a b l e s u i t o f c lo t h e s i n th e bushes b e f o r e r i d i n g i n t o town. S t e p h e n s had b e e n a l a w y e r f o r a week when he made t h e a c q u a i n t a n c e o f R o b e r t Toombs i n Washington c o u r t. The p r o f e s s i o n a l c o n n e c t i o n t h u s blossom ed i n t o a f a s t f r i e n d s h i p between t h e two. exceptions, i t was t o b e t h e f o r m e d s oo n With one o r two c lo s e s t frie n d sh ip o f Stephens' life. 28 27 P l e a s a n t A. S t o v a l l , Rober t T o o m b s : S t a t e s m a n , S p e a k e r , Sol d i e r , Sage . . . (New Y o r k , 1 8 9 2 T 7 1 5 - 1 6 ; Von A b e l e , S t e p h e n s , 55. 28 Toombs o f biography o f Robert character D e t a i l s o f Toombs' e a r l y l i f e i n W i l l i a m Y. Th ompson, R o b e r t G e o r g ia (Baton Rouge, 1 9 6 6 ) , 3 -1 2. Thompson's i s t h e l a t e s t o f Toombs. The b e s t i s s t i l l U l r i c h B o n n e r P h i l l i p s , L i f e Toombs (New Y o r k , 1 9 1 3 ). A l s o u s e f u l f o r a s t u d y o f Toombs' is S tovall, supra. 5^ Toombs was two y e a r s o l d e r t h a n S t e p h e n s. fifth He was b o r n t h e son o f a w e l l - t o - d o p l a n t e r in W ilkes c ounty i n 1810. contemptuous o f a l l U tterly a u t h o r i t y , Toombs h a d b e e n e x p e l l e d f ro m F r a n k l i n College a f t e r a s e r i e s of serious in fr a c tio n s o f the r u le s , including s w e a r i n g a nd a t t a c k i n g a p a i r o f f e l l o w s t u d e n t s w i t h a c l u b a nd pisto l. He w e n t on t o g r a d u a t e a t Uni on C o l l e g e i n S c h e n e c t a d y , New Y o r k , a n d t h e n h e s p e n t a y e a r a t V i r g i n i a Law S c h o o l , l e a v i n g t h e r e befo re o b ta in in g a degree. He h a d b e e n a p r a c t i c i n g l a w y e r f o r f o u r y e a r s when h e met S t e p h e n s. The r e l a t i o n s h i p b e t w e e n t h e s e two men p r o v e s t h e o l d saw t h a t opposites a t t r a c t. Toombs was e v e r y t h i n g t h a t S t e p h e n s was n o t. A b i g man, s t r o n g a nd h e a l t h y , h i s h u g e h e a d e l e g a n t l y t o p p e d w i t h g r e a t s h o c k s o f d a r k bro wn h a i r , Toombs was g r e g a r i o u s , quick-tongued, p a s sio n a te , and r i c h. w i t h b o t h h a n d s a nd r o a r e d i n i t s tero u s, profane, im pulsive, He was a man who g r a b b e d l i f e face i f i t dared to p r o t e s t. B ois and g i v e n t o i n t e m p e r a n c e - ~ h e h a d an e n d u r i n g f o n d n e s s f o r a l c o h o l i n j u s t a b o u t any o f i t s was as s k e p t i c a l o f r e l i g i o n a l l u r i n g m a n i f e s t a t i o n s — Toombs as S t e p h e n s was p i o u s , as c o n v i v i a l and h e a r t y a s S t e p h e n s was m o r o s e a n d i n t r o s p e c t i v e , a n d he was a s g i v e n t o m e l a n c h o l y as B a c c h u s him self. Bob Toombs, i n s h o r t , w o u l d seem t o b e t h e l a s t man on e a r t h t o be com p atib le w ith S te p h e n s. But i t so happened t h a t t h e i r c h a r a c t e r s f o r m e d an a l m o s t p e r f e c t m e s h. They w e r e o p p o s i n g —y e t life forces. T h a t was S t e p h e n s a n d Toombs. d o m in e e r i n g , and o u t g o i n g ; studious. com plim entary— The one i m p e r i o u s , t h e o t h e r c o n t r o l l e d , i n t r o s p e c t i v e , and T h e i r r e l a t i o n s h i p , b o t h p o l i t i c a l a n d p e r s o n a l , became 55 a legend. O t h e r t h a n h i s w i f e , upon whom Toombs b e s t o w e d a s o l i c i t o u s and g e n t l e a f f e c t i o n t h a t b e l i e d h i s g r u f f , b l u s t e r y e x t e r i o r , S tephens was t h e o n l y p e r s o n i n t h e w o r l d c a p a b l e o f i n c l u e n c i n g Bob Toombs. T h e i r d i s a g r e e m e n t s f o r o x e r f o r t y y e a r s c o u l d b e n u m b e r e d c n one hand. Toombs, w r o t e a s e n s i t i v e o b s e r v e r , " l o v e d A l e x a n d e r H. S t e p h e n s w i t h a t e n d e r n e s s t h a t was a l m o s t p a t h e t i c a nd was a s much b e l o v e d by him . . . a n d , i n a l l m a t t e r s o f i m p o r t a n c e , Mr. Toombs came u p , i n t h e e n d , on Mr. S t e p h e n s ’ s s i d e . " Like h i s 29 f r i e n d Toombs, S t e p h e n s was a g o o d l a w y e r. By t h e e a r l y l&UOs t h e two w e r e u n q u e s t i o n a b l y t h e t e s t l a w y e r s on t h e n c r t h e r n circu it. The way t h a t t h e s y s t e m o p e r a t e d — t h e r e was n o a p p e a l from a s u p e r i o r c o u r t j u d g m e n t — p l a c e d a h i g h premium on t h e o b t a i n i n g o f jury v e rd ic ts. This in tu r n r e q u i r e d o f advocates b e f o r e th e b a r e l o quence and p e r s u a s i v e n e s s. in abundance. And t h e s e q u a l i t i e s Stephens p ossessed F o r t h e m o s t p a r t , h i s m an n e r was t o c o n v i n c e j u r i e s by l o g i c a l e x p o s i t i o n o f t h e p r i n c i p l e s he q u i c k l y g r a s p e d. involved in th e case. But S t e p h e n s was a p o w e r f u l a d v o c a t e b e c a u s e he knew j u s t when t o a p p l y t h e o t h e r t o o l s o f h i s t r a d e. fiery These He c o u l d be a n d p a s s i o n a t e when t h e o c c a s i o n d e m a n d e d , e m p l o y i n g s a t i r e ev o k in g sympathy f o r h i s clien t or t o sway t h e e m o t i o n s o f b e n c h and jury alik e. By a l l speaker. a c c o u n t s S t e p h e n s was an a r r e s t i n g a n d c o m p e l l i n g p u b l i c Hi s v o i c e was t h e f i r s t thing to s tr ik e a l i s t e n e r. I t was ^ 9 v a r i n a H o w e ll D a v i s , J e f f e r s o n D a v i s , Ex- P r e s i d e n t o f t h e C o n f e d e r a t e S t a t e s o f A m e r i c a : A Memoir b y His W i f e , 2 v o l s . (New Y o r k , 1 8 9 0 ) , I , it l O - 1 1. 56 high-pitched, alm ost f e m in in e , could p la y i t like a ly re. " l e t us r e a s o n t o g e t h e r " a n d when S t e p h e n s was i n t o p f o r m , he Now low a n d s o f t i n a c o n v e r s a t i o n a l t o n e , now l o u d , p i e r c i n g , and p e n e t r a t i n g , w ith words and p h r a s e s b e i n g p r o p e l l e d a t t h e a u d ie n c e l i k e r i v e t s. Nor was h i s v o i c e S t e p h e n s ' m o st e f f e c t i v e t o o l a s an o r a t o r. one e v e n b e t t e r , He h a d an u n c a n n y a b i l i t y t o g u a g e t h e q u a l i t y a n d t e m p e r o f an a u d i e n c e a n d t o a d j u s t h i s m a n n e r a n d a r g u m e n t a c c o r d i n g l y. W h e r e e v e r h e s p o k e — on t h e s t u m p , i n a c o u r t r o o m , o r on t h e f l o o r o f C o n g r e s s — S t e p h e n s commanded u n d i v i d e d a t t e n t i o n . ^ How much in com e S t e p h e n s d e r i v e d fro m h i s d i f f i c u l t to say. in come f o r a l a w y e r. In s h o r t , a y e a r was a " l a r g e " a man w o u l d n o t g e t r i c h p r a c t i c i n g Large fee s were t h e e x c e p tio n r a t h e r than t h e r u l e : planters, is R i c h a r d J o h n s t o n , who was a d m i t t e d t o t h e b a r i n 181+3, s a y s t h a t t h r e e o r f o u r t h o u s a n d d o l l a r s law. law p r a c t i c e to avoid th e h a s s le s the w e a lth ie s t a n d d e l a y s an a p p e a l t o l aw e n t a i l e d , r e s o r t e d m o st o f t e n t o o u t - o f - c o u r t a r b i t r a t i o n by m u t u a l f r i e n d s. Bu t t h e l e s s p r o m i n e n t h a d no s u c h c o n v e n i e n c e s , a n d t h e l e g a l b u s i n e s s was a s t e a d y one a t l e a s t , n o t s u b j e c t t o t h e v i c i s s i t u d e s o f t h e season l i k e p la n tin g. And c i t y p r a c t i c e k e p t a g oo d l a w y e r b u s y. " N e a r l y e v e r y c i t i z e n o f p r o m i n e n c e i n t o w n , " w r o t e a Columbus l a w y e r , 30 R i c h a r d M a lc o l m J o h n s t o n , A u t o b i o g r a p h y o f C o l . R i c h a r d M a lc o lm J o h n s t o n ( W a s h i n g t o n , 1 9 0 0 ) , 1.13, lU2-i+3 W itnesses never f a i l e d t o remark S t e p h e n s ' manner o f s p e a k i n g , and a lm o s t a l l a t t e s t e d t o th e a t t e n t i o n a S te p h en s' speech com pelled. Even f o r e i g n e r s who u n d e r s t o o d n o t a wor d o f E n g l i s h w e r e m e s m e r i z e d b y S t e p h e n s ' o r a t o r y. New York T r i b u n e , 15 F ' e b r u a r y 1 8 5 9 , q u o t e d i n R a b u n , " S t e p h e n s , " 595. 57 "was a d e f e n d a n t i n o n e o r more s u i t s . " 31 I t is quite possible th at t h e p r a c t i c e o f a s u p e r i o r l a w y e r l i k e S t e p h e n s y i e l d e d him more t h a n th e normal revenue. Ho w ev er, t h e s e p r o s p e r o u s d a y s w e r e s t i l l h o t summer m o n th s o f J u l y a n d A u g u s t , 1 8 3 ^. f i n d i n g i t s way t o h i s in the fu tu re in the So l i t t l e b u s i n e s s was c o u rth o u se o f f i c e t h a t Stephens began t o th in k a b o u t mov ing w e s t t o A la b am a o r M i s s i s s i p p i. The e n t i r e month o f A u g u s t n e t t e d h im one p r o m i s s o r y n o t e f o r t h e m a g n i f i c e n t sum o f t w e n t y five d o llars. literally , And t h e f i r s t h a r d c a s h h e r e c e i v e d f o r h i s s e r v i c e s — f o r i t was f o u r s i l v e r h a l f - d o l l a r s — was f o r s p e a k i n g i n c o u r t f o r one Ja me s F a r m e r on 1 S e p t e m b e r. O t h e r t h a n d r a w i n g up a few l e g a l d o c u m e n t s , S t e p h e n s h a d a c c o m p l i s h e d n o t h i n g as a l a w y e r. He c e r t a i n l y d i d n o t l a c k t h e t i m e t o w r i t e i n q u i r i n g l e t t e r s in newer p o r t i o n s of the s ta t e. Was t h e r e p e r h a p s a f u t u r e to friends fo r a sharp y o u n g l a w y e r o u t t h e r e 7^^ Then came a b r e a k. purchased h is On 10 S e p t e m b e r , t h e same day t h a t S t e p h e n s f i r s t h o r s e — he c o u ld s t a n d t h e em barrassm ent o f b o rro w in g o n e no l o n g e r — a man by t h e name o f Ja me s H i l s m a n came t o t h e o f f i c e and employed h i s d rifter, services. b u t h e n e e d e d a l a w y e r a n d p r o m i s e d t o pa y t w e n t y d o l l a r s. S t e p h e n s , upon h e a r i n g h i s The s t o r y was t h i s : copy in s to r y , took the case. H i l s m a n ' s w i f e , n e e Amanda Askew, h a d i b i d . , 103; SHC/NC, k 2. 31johnston, scrip t H i l s m a n was a n e ' e r - d o - w e l l , a b e s o t t e d " A utobiography of R. J . M o s e s , " type 3^AHS t o L S , 20 A u g u s t 1 8 3 ^ , S t e p h e n s P a p e r s , MC; AHS D i a r y , 7 S e p t e m b e r 183^+, i b i d . , LC: Ja me s W. E'sby t o AHS, 27 S e p t e m b e r 1 8 3 ^ , ibid. 58 b e e n a widow when h e m a r r i e d h e r. Her f i r s t h u s b a n d , a man named Uriah B a t t l e , had d ied s h o r t l y a f t e r the b i r t h o f a d au g h te r. father, a w e a l t h y Hancock c o u n t y p l a n t e r , im m ediately took out l e t t e r s o f g u a r d i a n s h i p on t h e p e r s o n a n d p r o p e r t y o f t h e i n f a n t. Hilsman s t a g g e r e d i n t o t h e p i c t u r e the e ld e r B a ttle B a ttle 's and m a r r i e d Amanda. Then N aturally, c o u l d n o t l o o k upon t h i s t u r n o f e v e n t s w i t h e q u a n i m i t y , a n d on t h e b a s i s o f h i s letters o f g u a r d i a n s h i p he had t h e c h i l d s e i z e d f r o m i t s m o t h e r a n d c a r r i e d t o h i s home. Hilsman t h e n a p p e a le d t o t h e law t o r e c o v e r t h e c u s to d y o f h i s w i f e ' s child. The c a s e e x c i t e d an e x t r a o r d i n a r y amount o f i n t e r e s t i n T a l i a f e r r o and t h e s u r r o u n d i n g c o u n t i e s. and i n f l u e n t i a l f a m ily. The B a t t l e s w e r e an e x t e n d e d P u b lic o p in io n overw helm ingly sym pathized w i t h th em a n d t h e i r s i d e o f t h e c a s e. When t h e c a s e f i n a l l y w e n t t o trial, some o f t h e m fro m t w e n t y o r a l a r g e crowd o f a r e a c i t i z e n s , t h i r t y m iles o f f ( f o r t h a t t i m e w h a t am o u n t e d t o a w h o l e d a y ' s j o u r n e y ) , was on h a n d. The B a t t l e s ' l a w y e r was no ne o t h e r t h a n S w e p s t o n J e f f r i e s , S t e p h e n s ' m entor and a h i g h l y r e g a r d e d a t t o r n e y. But i t was t h e y o u n g e r l a w y e r who w o u l d p r e v a i l t h a t d a y. A l t h o u g h h e was unknown t o n i n e - t e n t h s o f t h e c o u r t r o o m c r o w d , the p a le , t r e m b l i n g , a n d g h a s t l y - t h i n y o u n g man w i t h t h e p i p i n g v o i c e soon t r a n s f i x e d them w i t h h i s e lo q u e n c e. His t he m e was t h e s a n c t i t y o f m o t h e r h o o d , t h e l aw o f n a t u r e t h a t r e q u i r e d e v e n f o r e s t b r u t e s f i g h t to th e death fo r th e s a f e ty o f t h e i r o f f s p r in g. h a d f i n i s h e d , m o st o f t h e c o u r t r o o m , we a r e t o l d , to By t h e t i m e he including the five 59 judges were in t e a r s. drifter; F o r g o t t e n was H i l s m a n , t h e g o o d - f o r - n o t h i n g f o r g o t t e n were th e B a t t l e s ' f o r t u n e and r e p u t a t i o n , t h e w h e re w ith a ll t o provide s p le n d id ly fo r th e c h i l d 's u p b rin g in g. B a ttle 's l e t t e r s were s e t a s i d e. The m o t h e r r e g a i n e d c u s t o d y o f h e r c h i l d , a n d t h e c a s e made S t e p h e n s ' E v id en tly , though, th is r e p u t a t i o n .^3 r e p u t a t i o n d i d n o t grow f a s t en o ug h t o s u i t h i m , f o r he c o n t i n u e d t o i n v e s t i g a t e t h e p o s s i b i l i t i e s m ov ing o u t o f C r a w f o r d v i l l e. of Hi s i n t e r e s t i n p o l i t i c s was s t e a d i l y gro w in g , and f o r a w h i l e he to y e d w ith t h e i d e a o f g o in g i n t o new spaper w o r k , w r i t i n g A. B. L o n g s t r e e t o f t h e A u g u s t a S t a t e R i g h t s S e n t i n e l about p o s s ib le openings. But, r e s t l e s s as he w a s , S te p h e n s by t h i s t i m e h a d become t o o a t t a c h e d t o C r a w f o r d v i l l e s e r i o u s l y t o c o n s i d e r leaving i t. I t was a b o u t t h i s t i m e t h a t J e f f r i e s o f f u l l p a r t n e r s h i p i n Co lu m b u s. He r e f u s e d. He now h a d s e t h i s h e a r t n o t o n l y on s t a y i n g , b u t a l s o on b u y i n g b a c k h i s when t h e o p p o r t u n i t y a r o s e. made him t h e o f f e r fa th e r's The " h o o k s o f s t e e l , " old property as h e l a t e r c a l l e d t h e m , t h a t b o u n d h im t o C r a w f o r d v i l l e w e r e a l r e a d y f i r m l y embedded a f t e r o n l y a few m o n th s i n h i s c h ild h o o d home."^ I t was now 1835 a n d S t e p h e n s was malting a l i v i n g. Business ■ ^ J o h n s t o n a nd Br o wn e , S t e p h e n s , 9 5 - 9 7.	22,347
https://openalex.org/W2914278961	OpenAlex	Open Science	CC-By	2,019	Qualitative Interview Studies of Working Mechanisms in Electronic Health: Tools to Enhance Study Quality	Marianne Therese Smogeli Holter	English	Spoken	9,693	16,035	Qualitative Interview Studies of Working Mechanisms in Electronic Health: Tools to Enhance Study Quality Marianne TS Holter1, MA; Ayna B Johansen1, PhD; Ottar Ness2, PhD; Svend Brinkmann3, PhD; Mette T Høybye4,5, PhD Hå B d 1 PhD Marianne TS Holter1, MA; Ayna B Johansen1, PhD; Ottar Ness2, PhD; Svend Brinkmann3, PhD; Mette T Høybye4,5, PhD; Håvar Brendryen1, PhD 1Norwegian Centre for Addiction Research (SERAF), Institute of Clinical Medicine, University of Oslo, Oslo, Norway 2Department of Education and Lifelong Learning, Norwegian University of Science and Technology (NTNU), Trondheim, Norway 3Department of Communication and Psychology, Aalborg University, Aalborg, Denmark 4Elective Surgery Centre, Regional Hospital Silkeborg, Silkeborg, Denmark 5Department of Clinical Medicine, Interacting Minds Centre, Aarhus University, Aarhus, Denmark *all authors contributed equally Corresponding Author: Marianne TS Holter, MA Norwegian Centre for Addiction Research (SERAF) Institute of Clinical Medicine University of Oslo Postboks 1039 Blindern Oslo, 0315 Norway Phone: 47 93623061 Email: marianne.holter@protonmail.com Corresponding Author: Marianne TS Holter, MA Norwegian Centre for Addiction Research (SERAF) Institute of Clinical Medicine University of Oslo Postboks 1039 Blindern Oslo, 0315 Norway Phone: 47 93623061 E il i h lt @ t il JOURNAL OF MEDICAL INTERNET RESEARCH JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al Abstract Future development of electronic health (eHealth) programs (automated Web-based health interventions) will be furthered if program design can be based on the knowledge of eHealth’s working mechanisms. A promising and pragmatic method for exploring potential working mechanisms is qualitative interview studies, in which eHealth working mechanisms can be explored through the perspective of the program user. Qualitative interview studies are promising as they are suited for exploring what is yet unknown, building new knowledge, and constructing theory. They are also pragmatic, as the development of eHealth programs often entails user interviews for applied purposes (eg, getting feedback for program improvement or identifying barriers for implementation). By capitalizing on these existing (applied) user interviews to also pursue (basic) research questions of how such programs work, the knowledge base of eHealth’s working mechanisms can grow quickly. To be useful, such interview studies need to be of sufficient quality, which entails that the interviews should generate enough data of sufficient quality relevant to the research question (ie, rich data). However, getting rich interview data on eHealth working mechanisms can be surprisingly challenging, as several of the authors have experienced. Moreover, when encountering difficulties as we did, there are few places to turn to, there are currently no guidelines for conducting such interview studies in a way that ensure their quality. In this paper, we build on our experience as well as the qualitative literature to address this need, by describing 5 challenges that may arise in such interviews and presenting methodological tools to counteract each challenge. We hope the ideas we offer will spark methodological reflections and provide some options for researchers interested in using qualitative interview studies to explore eHealth’s working mechanisms. (J Med Internet Res 2019;21(5):e10354) doi: 10.2196/10354 J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 1 (page number not for citation purposes) The Need to Identify the Working Mechanisms of Automated Electronic Health Programs The qualitative interview is also a pragmatic research method, as many researchers already conduct interviews with program users as part of an applied research goal (developing or implementing an intervention). In the process of conducting interviews with program users, a researcher may become intrigued by a more basic research question and may perhaps consider the pragmatic solution of pursuing both the applied and the basic research goal in the same interviews by simply adding questions to the existing interview guide. We believe that such studies mixing basic and applied research goals have the potential of becoming an important asset to the field, by accumulating knowledge on more general issues that may help us understand how eHealth therapy works. Building the next generation of automated electronic health (eHealth) programs will require a shift of attention from the performance of individual programs to a joint effort of understanding eHealth’s working mechanisms [1,2]. The term eHealth is a broad term that may refer to many forms of technological health support; this paper concerns itself with automated Web-based interventions for mental and physical health purposes [3], or eHealth programs. The outcomes of individual eHealth programs are well established; a vast majority of the research published between 1996 and 2013 concerned outcome (74%). However, much fewer publications focused on processes (26%) [4]. Consequently, the working mechanisms that underlie the outcomes of individual interventions are substantially less documented [5-7]. This is not only theoretically unsatisfactory; it is also problematic when it comes to designing new eHealth programs, as there are few, if any, field-specific theories of models that can be used to inform program development. However, to become such an asset, the interviews conducted in these studies should provide what in qualitative methodology is known as thick descriptions or rich data [20,21]. Rich data are usually considered a requirement for a valid qualitative analysis, and the concept signifies having enough data of sufficient quality relevant to the research question, including both variation (ie, data breadth) as well as details and nuances (ie, data depth) [22,23]. Data that are not rich—that lack in breadth or depth—might threaten the study’s quality or the potential reach of its conclusions. Thus, getting rich data that inform the research question is an important aspect of a qualitative study. KEYWORDS th; mobile health; telehealth; mHealth; interviews as topic; health care evaluation mechanisms; data collectio telemedicine; eHealth; mobile health; telehealth; mHealth; interviews as topic; health care evaluation mech telemedicine; eHealth; mobile health; telehealth; mHealth; interviews as topic; health care evaluation mechanisms; data collection J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 1 (page number not for citation purposes) https://www.jmir.org/2019/5/e10354/ XSL•FO RenderX JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al for uncovering important insights about the processes that may be involved in automated eHealth therapy [16-19]. The Need to Identify the Working Mechanisms of Automated Electronic Health Programs In the case of qualitative interviews, producing rich data means conducting interviews in a way that makes the participants spend a lot of time talking about the aspects that are central to the investigation, including both breadth and depth in their descriptions. This may seem straightforward, but it can be surprisingly difficult in practice. Instead, the development of eHealth programs often relies on rather static traditional behavior change theories [2] or models from face-to-face therapy [8-11], under the assumption that the principles are transferrable to automated eHealth therapy. However, the interaction between a program user and an automated eHealth program is in many cases not static; many programs include different degrees of interactivity and tailoring [3,5], making traditional behavior change theories potentially unsuitable [1,2]. On the other hand, using models from face-to-face therapy may not be appropriate either, as automated eHealth therapy by definition does not involve human contact. As automated eHealth programs are neither static nor involve human contact, it is possible (or even likely) that the way they achieve their effects is not explained with established theories and models [1,2]. This suggests a need for research that can identify eHealth’s working mechanisms, knowledge on which it is possible to build eHealth-specific theories and models. https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 2 (page number not for citation purposes) JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al behavior change processes (Figure 1, adapted from Moen and Middelthon’s discussion of interviews) [34]. A health intervention’s working mechanisms can be conceptualized as how the interaction between the person and the health intervention influences the person’s internal change processes. The interaction, in turn, can be described as a combination of the interaction’s content (the what of the interaction) and the interactional processes (the how of the interaction). For example, psychotherapy’s working mechanisms can be described as the therapy sessions’ influence on the client’s internal change processes. The therapy sessions, in turn, can be described as comprising 2 main elements: their content (eg, the topic discussed) and the interactional processes, when and how often interaction is initiated, how the interaction unfolds, how the next interaction is initiated, and so on. We opted for trying to improve the interview method; however, we found no guidelines within the field of eHealth for how to conduct high-quality qualitative interview studies on potential eHealth working mechanisms. Therefore, we started looking more closely at the interviews we had conducted, asking ourselves what had gone wrong. This process led to the identification of a handful of problems that we believed were likely to have contributed to the difficulties in getting rich data on how the participants related to the program. As we started defining these problems, we discovered that we had also encountered several of them in other eHealth studies we had been involved in [29-33], and we therefore believed they could be relevant beyond the specific study we were currently engaged in. We wanted our experiences to be of benefit to other researchers with similar agendas and interests, and we therefore sought to describe the problems we had encountered in a way that would maximize their generalizability. Thus, through discussion among ourselves and with other researchers, we conceptualized 5 interview challenges: achieving a joint understanding of the interview topic, keeping participants from straying off the focus of enquiry, aiding recall of specific program experiences, avoiding negative influence of the social interview situation, and structuring the dual-aim interview. Having identified the challenges, we consulted the literature on qualitative methodology to identify methodological tools to counteract each challenge. However, interventions may differ according to how much the 2 interacting parties—the person and the intervention—influence the interactional content and the interactional processes. JOURNAL OF MEDICAL INTERNET RESEARCH In the case of psychotherapy, both the client and the therapist highly influence both components of the interaction. Taking another example, a person reading a self-help book is also interacting with a health intervention: things also act, and people interact with them, in that the properties of a thing influence how a course of action involving that thing unfolds [35]. The working mechanisms of a self-help book can therefore also be described in terms of the interactional content and the interactional processes. However, the relative influence of the 2 interacting agents (reader and book) differ from the case of psychotherapy. The book decides the interactional content, although the reader largely decides the interactional processes: when and how often interaction is initiated (when to read again), how the interaction unfolds (what to read in what sequence), how the next interaction is initiated (picking up the book), and so on. Returning to the study that had started this process [24], we changed the interview method to include some of the tools we had identified. This markedly enhanced the quality of subsequent interviews, producing rich data to answer how the participants related to the eHealth program as well as whether this way of relating influenced change. Thus, although the original interview method generated scant data on the basic research question, the revised interview methodology led to interviews that could answer the same research question with rich data. Considering the working mechanisms of an eHealth program, many programs will influence both the interactional content and the interactional processes. As with a self-help book, the interactional content will usually to a large extent be decided by the program. Moreover, just as a self-help book, the program is a thing, and many people are likely to think of things such as computer programs as inanimate objects with content. Indeed, the most prominent feature of eHealth programs is their content [3,10], even though they also may substantially influence the interactions with the user [3]. For example, eHealth programs may influence when and how often the interaction takes place (eg, through reminders to log on), how the interaction unfolds (eg, by responding with tailoring to user input), how the next interaction is initiated (eg, through invitation), and so on [5,10]. JOURNAL OF MEDICAL INTERNET RESEARCH Some of these interactional processes may not be experienced directly by the individual user, for example, in the case of tailoring, the program may be adapted specifically to the user’s input, but she or he nevertheless only sees 1 version of the program, masking the actual interaction. In sum, although eHealth programs may have a substantial influence on the interactional processes, the average program users may primarily focus on their content and think of them as inanimate objects that do not interact. In other words, to the user, the interaction with the program can be largely invisible (Figure 2). In short, although it seemed a pragmatic solution to use already-planned interviews to pursue the answer to a basic eHealth research question, we experienced that getting rich data on the basic research question was challenging. In the absence of guidelines for conducting high-quality qualitative interviews specifically adapted to the field of eHealth, the process we entered into led to an enhanced methodological awareness and specific methodological tools for increasing study quality. The main focus of this paper is to share the identified challenges and tools with the research community. However, before doing so, we will offer what we consider to be a handy heuristic for understanding some of these methodological challenges: the invisible interaction between eHealth program and program user. https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 3 (page number not for citation purposes) Difficult in Practice: The Case of a Study on a Person-To-Program Alliance The reflections that are presented in this viewpoint paper arose from some of the authors’experiences with a specific interview study [24] (in review), the aim of which was to explore a potential person-to-program alliance. The study in question included the development of an alliance-supporting program [25], and in an early study phase, the interviews had both an applied and a basic purpose: the applied purpose was getting feedback for program improvement, and the basic purpose was exploring how the participants related to the program. By exploring how participants related to the program, we hoped to achieve a better understanding of a person-to-program alliance [26-28] as a potential eHealth working mechanism. However, it was surprisingly difficult to conduct interviews that would yield rich data on how the participants related to the program, and the initial interviews resulted in scant data to answer the research question. This left us with 3 options: (1) answering the basic research question with scant data, which would limit the conclusions we could draw, (2) abandoning the basic research question as unanswerable, or (3) trying to generate richer data by changing the way the interviews were being conducted. Qualitative Interviews: A Promising and Pragmatic Method for Studying Electronic Health Programs’ Working Mechanisms eHealth’s working mechanisms can be studied using various methods, but a promising and pragmatic venue of investigation is the qualitative interview, that is, “professional conversations (...) where knowledge is constructed in the inter-action between the interviewer and the interviewee (...) about a theme of mutual interest” [12]. The qualitative interview is a promising method for investigating eHealth working mechanisms as it grants unique access to participants’experiences and as it is especially suited to explore what is unknown [13]. Providing a means to explore the unknown makes qualitative interviews a potent method for generating new knowledge and theory [14,15], and some interview studies have already demonstrated their potential J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 2 (page number not for citation purposes) XSL•FO RenderX JOURNAL OF MEDICAL INTERNET RESEARCH A Handy Heuristic: The Invisible Interaction We suggest that a person’s interaction with any health intervention can be visualized in terms of a triangle, which includes the individual help seeker, the intervention, and the J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 3 (page number not for citation purposes) https://www.jmir.org/2019/5/e10354/ XSL•FO RenderX Achieving a Joint Understanding We believe it is a truism that experience is multifaceted and that an experience can be described from many perspectives. For example, a client may describe a therapy session from a factual perspective of when and where it took place, from an experiential perspective of his or her emotions before, during, and after the session, from a historical perspective of the session as a stage in his or her spiritual development, and so on. If the interviewer’s questions are mostly descriptive, there may be a scarcity of cues concerning which perspective to assume, leaving the decision up to the participant—and the participant’s choice may not be the researcher’s choice. This may be especially challenging in studies on eHealth working mechanisms, as the interactional processes may not be part of the participant’s conscious experience. Therefore, descriptive questions asking for the participant’s program experiences will perhaps not cause him or her to talk about the (invisible) person-program interaction but rather about the program as a thing with a content. On the other side of the conversation, the interviewer may fear that more direct questions onto the focus of enquiry will put words in the participant’s mouth and disqualify any subsequent answer. A final tool to clarify and exhaust the interview topic is to ensure the possibility of conducting follow-up interviews [41]. A follow-up interview gives both the interviewer and the participant an opportunity to reflect on what was talked about in the first interview, allowing new insights or aspects to emerge [22]. It also gives the researcher an opportunity to clarify questions or test interpretations with the participant directly [39], giving more nuanced data and enhancing validity. Coresearcher design and vignettes foster clearer communication, but they may also threaten the study’s validity if the researcher holds on to his or her initial assumptions about the studied process, failing to acknowledge unexpected perspectives. To ensure that these tools strengthen and not hamper the quality of the study, the researcher should adopt what in psychotherapy is known as the beginners mind: remaining curious and receptive, open to all possibilities [42-44]. Furthermore, the interviewer must throughout the research process practice reflexivity, that is, considering how she or he may be affecting the study with “(…) thoughtful, conscious self-awareness” [44]. Reflexivity about, for example, preunderstandings, motivations, and the influence of previous experiences can lead to important insights [43,44]. Achieving a Joint Understanding When a researcher sets out to explore a potential eHealth working mechanism in an interview, it may be difficult to achieve a joint understanding of the interview topic together with the participant. For example, as mentioned previously, in the study that was the starting point for this paper, the researchers were interested in understanding how the participants related to the program [24]. The interview guide comprised mainly of descriptive interview questions—questions that ask the participant to describe a specific experience, which are usually recommended to get close to the participants’ own experience [13]. Examples of descriptive interview questions were could you tell me what you thought and felt the first time you used the program and could you describe the role the program has had in your quit attempt (the program was for helping people quit smoking). However, the researchers struggled with superficial answers that did not seem to reveal anything about how the participants related to the program (such as I thought the program was fine) until 1 participant called the program a secret friend. The researchers were puzzled. Was this person’s program experience unique? Why did other participants not talk about the program in this way at all? However, apart from a few statements similar to this one, the interviews were not generating data to answer the research question of how the participants related to the program. Another and more direct way of fostering a joint understanding of the interview topic is to involve the participants as coresearchers, or using epistemic interviewing [13,14,39,40]. In traditional qualitative interviewing, participants describe their subjective experiences and the data are analyzed and interpreted afterward by the researcher [13,14]. In contrast, a coresearcher design entails that the researcher shares his or her current understanding of the research topic and asks for the participant’s views, and the research questions are investigated in collaboration. Involving participants as coresearchers also changes the roles of the interviewer, who becomes a sort of participant contributing with his or her perspective. This joint exploration entails that much of the analysis and validation is done in the interview [14,39]. Failure to get rich data on a research question may indicate a marginal phenomenon—or that the interviewer is failing to communicate the focus of enquiry in a way that facilitates joint understanding with the participant. JOURNAL OF MEDICAL INTERNET RESEARCH JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al her attention toward them. In other words, being largely invisible, the interactional processes may not be part of the participant’s conscious experience that she or he is ready to share in an interview. This may create or contribute to certain challenges with exploring eHealth working mechanisms through interviews. We will now present 5 such challenges and suggest methodological tools to counteract them. The invisible interaction is a useful heuristic when considering the challenges of interview studies for exploring eHealth’s working mechanisms. We previously stated that an asset of qualitative interviews is their potential to explore eHealth working mechanisms from the program user’s perspective. However, from this perspective, part of the program’s working mechanisms—the interactional processes— are maybe invisible to the participant, unless she or he purposefully directs his or Figure 1. Working mechanisms of a behavior change intervention. Figure 1. Working mechanisms of a behavior change intervention. Figure 2. Working mechanisms of an automated electronic health intervention. Figure 2. Working mechanisms of an automated electronic health intervention. Figure 2. Working mechanisms of an automated electronic health intervention. Figure 2. Working mechanisms of an automated electronic health intervention. https://www.jmir.org/2019/5/e10354/ https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 4 (page number not for citation purposes) https://www.jmir.org/2019/5/e10354/ JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al guide. The vignette can be constructed on the basis of a participant account, on relevant literature, or on the researcher’s current understanding of the processes under study. The interviewer might introduce the vignette by saying that she or he wants to share a story with the participant. After recounting the vignette, the interviewer can ask for the participant’s reactions and ask follow-up questions (eg, if the participant has experienced anything similar or can provide a different perspective). Using several vignettes in the same interview can be a useful way of illustrating different perspectives on the research topic. This will implicitly communicate to the participant that all answers are acceptable, ensuring that the vignettes function to guide the conversation but not restrict the answers [36,38]. Achieving a Joint Understanding Reflexive insights that could be of importance for analysis should be documented (eg, through memos or notes) [15,43,44]. Finally, documented reflections should be made part of the analysis and be made explicit to the reader [44]. An interviewer can use several methodological tools to foster a joint understanding of the interview topic with the participant. One such tool is vignettes: vivid, exemplifying prose stories that guide the conversation toward a particular aspect of the participant’s experience [36-38]. The interview vignette is constructed before the interviews and included in the interview https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 5 (page number not for citation purposes) Keeping Participants From Straying off the Focus of Enquiry To allow time for joint exploration of the person-program interaction, it is necessary to limit the interview time spent on XSL•FO RenderX XSL•FO RenderX XSL•FO RenderX JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al To her surprise, several participants who were still active program users and had completed most sessions up until the time of the interview had difficulties remembering any particular program session at all. matters that are not at the core of the research question. Returning to Figure 2 and the triangle of program, user, and behavior change, the relative importance of each triangle endpoint will vary according to the research question: some parts of the triangle will be the focus of enquiry, whereas the other parts will be contextual. For example, in 1 study [33] (paper under preparation), the researchers interviewed patients who had gambling problems and had used a Web-based referral site to connect with problem gambling services. The focus of enquiry was their use of the website; the gambling problems were the context. However, the interviewer struggled with keeping the conversation focused on the website, as participants talked mostly about their personal history with gambling problems. When asked about their experiences with the website, they appeared to feel alienated and at a loss. Consequently, there was a lot of interview data on the participants’behavior change efforts—but little data on their use of the website. matters that are not at the core of the research question. Returning to Figure 2 and the triangle of program, user, and behavior change, the relative importance of each triangle endpoint will vary according to the research question: some parts of the triangle will be the focus of enquiry, whereas the other parts will be contextual. For example, in 1 study [33] (paper under preparation), the researchers interviewed patients who had gambling problems and had used a Web-based referral site to connect with problem gambling services. The focus of enquiry was their use of the website; the gambling problems were the context. However, the interviewer struggled with keeping the conversation focused on the website, as participants talked mostly about their personal history with gambling problems. When asked about their experiences with the website, they appeared to feel alienated and at a loss. Consequently, there was a lot of interview data on the participants’behavior change efforts—but little data on their use of the website. Keeping Participants From Straying off the Focus of Enquiry Recalling specific program experiences may be challenging as although participants may be active program users at the time of the interview, they are not engaging with the program at that particular moment (unless you are combining the interview with a think-aloud-technique, discussed below) [45]. That means that to talk about program experiences, the participants must retrieve memories. However, program sessions may be short, and the participants are likely to use the program in between their other daily business. Consequently, program use may not be encoded as distinct episodic memories to begin with [46]; rather, these memories may be intertwined with other memories of everyday life. Thus, if the interviewer asks the participant to describe a program session, his or her question may not contain the right memory cues [47] to trigger memories of program use, and the participant may seemingly not recall any sessions at all. The invisible interaction may amplify this problem: if the participant is unaware of the program influencing the interaction, these program aspects will be even more difficult to retrieve on demand. If the participants continuously stray off the focus of enquiry by spending time on contextual aspects, it can threaten the data richness. Aspects that are contextual to the researcher may be aspects the participant wants to share or aspects she or he believes to be important to the investigation. The interviewer may try to lead the conversation back onto the focus of enquiry, but the participant may return to the contextual aspects, turning the interview into a battle over topic. Apart from being unpleasant for both, the result may be scant data on the focus of enquiry. When the focus of enquiry is potential eHealth working mechanisms, the invisible interaction may add to the challenge of straying off the topic. As the participant may be largely unaware of the interactional processes, she or he will instead talk about the aspects of which she or he is aware: the change processes (in isolation of the program) or the program (in isolation of the change processes). Information about the behavior change and about the program is certainly relevant contextual information, but talking about these aspects in isolation should not dominate the interview. There are, however, methodological tools to amend the problem with recall in the interview situation: 1 such tool is to get live access to the person-program interaction through the think-aloud procedure [45]. Keeping Participants From Straying off the Focus of Enquiry In the think-aloud procedure, the participants go through (parts of) the program during the interview as the interviewer instructs the participant to think aloud, reporting all thoughts without censoring them. The interviewer should not interrupt the participant’s flow of thoughts, and follow-up questions should be saved for after the think-aloud procedure is completed [45]. However, there are some limitations to this approach: unless the program comprises just 1 website or session, the researcher cannot use the think-aloud procedure to go through all program content, requiring him or her to select the most relevant sessions. Furthermore, when the focus of enquiry is working mechanisms within the invisible person-program interaction, the interviewer’s presence may draw attention from the program’s role in the interaction, adding to its invisibility. However, if these issues do not apply, the think-aloud procedure can enable a researcher to study possible eHealth working mechanisms as they happen, potentially removing the problem of recall. The interview conversation can be kept from straying off the research topic by using in-interview questionnaires to keep contextual answers short. The questionnaire can include questions addressing contextual issues (eg, How long have you been worried about your gambling? Or Have you tried restricting how much you gamble before?), together with any other questions that might serve as relevant analytic background (eg, demographics). The interviewer may fill out the questionnaire together with the participant at a suiting point during the interview. Using a piece of paper to fill out the answers will help keep the answers short, by providing limited space and communicating a wish for answers that the interviewer can write down. Short contextual answers will in turn leave more time for the focus of enquiry. Another tool for aiding recall is asking memory-facilitating interview questions. If program experiences have not been encoded as specific episodic memories, the interviewer’s phrasing of questions becomes increasingly important, as the words she or he uses will influence the participant’s memory-retrieval process by serving as memory cues [47]. The interviewer’s choice of words can be guided by mapping the participant’s program habits early in the interview. Knowledge of program habits can in turn be used to phrase questions in ways that contain memory cues; reflecting what the participant was doing before using the program, where she or he was, and his or her emotional state at the time of the experience [47]. Such memory-facilitating interview questions may help the https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 6 (page number not for citation purposes) Structuring the Dual-Aim Interview Gender stereotypes are not the only potential social disturbances in an interview—other social roles may be prominent, and within eHealth research, the interviewer may be particularly prone to be perceived as an interviewer or clinician or interviewer or developer. Perceiving the interviewer as also a clinician may cause the participant to think of him or her as a therapeutic interactional partner and to be less attentive to the therapeutic agency of the eHealth program. Similarly, perceiving the interviewer as also a program developer may highlight the program as a thing made by someone else, making it more difficult to see the program’s role as a therapeutic agent—or cause the participant to self-censor negative experiences, as 1 of the authors experienced in 2 different studies [29,31]. In both cases, the interviewer’s presence may cause the participant to think of the interviewer as the interacting agent, pushing the experience of the program as an interacting agent to the background and adding to the interaction’s invisibility. In sum, the social interview situation may cause the participant to talk differently about his or her program experiences than she or she would have otherwise. The consequence of this may be less rich data, or data that do not correctly represent the participant’s experience. It was mentioned in the introduction that qualitative interviews are pragmatic for exploring potential eHealth working mechanisms as the development or implementation of eHealth programs often entail user interviews anyway. Therefore, researchers who are interested in exploring potential eHealth working mechanisms may do so through existing interviews with applied purposes. However, when applied and basic research goals are mixed like this in the same interview study, it may create an additional challenge in getting rich data on the basic research question. In the study that inspired this paper [24], early interviews had both an applied research goal (getting feedback for improving the program) and a basic research goal (understanding how the participants related to the program as a potential eHealth working mechanism). The interview guide started with questions addressing possible sources for program improvement (participants’ likes or dislikes, specific program elements). Toward the end of the interview guide, questions on how the participants related to the program gradually increased in number (Has the program ever made you happy? Have you ever been upset by the program?). Aiding Recall of Specific Program Experiences Sometimes participants may not recall program experiences in sufficient detail to answer the interviewer’s questions. In the study that inspired this paper [24], the interviewer asked the participants to tell her about a program session they remembered especially well, thinking that she would use this session as a starting point for further descriptive interview questions [13]. XSL•FO RenderX JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al can make the interviewer appear as a clinician or a program developer, the interviewer may try to change these circumstances beforehand, for example, by changing the interview location or considering how to dress or talk. Alternatively, these issues can be addressed explicitly in the beginning of the interview, clarifying the interviewer’s role [19]. During the interview, the interviewer should try to monitor the social exchange [43], making notes of elements that may be impacting the conversation. After the interview, anything that might be of importance to the analysis should be documented [44]. These notes should be included somewhere easily accessible (eg, in the interview transcript or in a separate document) and analyzed as data that might inform, confirm, or qualify the analysis. Regarding the danger of additionally concealing the invisible interaction through the social exchange between the interviewer and the participant, the interviewer can try to arrange the interview situation so that it includes all 3 as potential agents: the participant, the program, and the interviewer [34]. participant disentangle the recall of program experiences from everyday life. As a final note on program recall, it may not be necessary for the participant to remember any particular program session at all; the researcher must consider what level of detail is necessary to answer the research questions meaningfully. For some research questions, the sum of program experiences may be more important than any particular experience. If so, using the interview to discuss the participant’s overall experience with the program can be more meaningful than facilitating recall of specific sessions [22]. Avoiding Negative Influence of the Social Interview Situation All interviews are also social situations, and aspects of the social situation will influence the data [48]. In 1 of the interviews from the study that inspired this paper [24], a female interviewer interviewed a male participant, with the goal of understanding how he related to the eHealth program he had used. The interview was brief and disappointing; the participant’s answers were short, and the topic was exhausted quickly. It was not until later that the interviewer became aware that she had been afraid of the participants judgment; that he would perceive her as a typical woman, valuing emotions (interactional processes; relating to the program) over facts (the program content). This subconscious fear had caused her to rush through the questions (which she during the interview had found awkward), partly answering some of them on behalf of the participant and ending the interview early. Finally, it is important to acknowledge that although the social interview situation may sometimes be a negative influence on the data, it can also be an asset. Through the interviewer’s reflexivity, the social situation may generate insights that would otherwise be missed. The interview in which the interviewer had rushed through the questions as she feared being labeled an emotional woman was considered as empirical material highlighting a possibly relevant aspect of how people relate to a program, namely, that relating to a program may go against social norms and produce feelings of embarrassment (in this case, as felt by the interviewer). https://www.jmir.org/2019/5/e10354/ J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 7 (page number not for citation purposes) Concluding Thoughts Conducting qualitative interviews is a promising and pragmatic approach for identifying the working mechanisms of automated eHealth programs. Existing user interviews for applied purposes can be used to also pursue basic research questions on eHealth working mechanisms. Researchers planning to conduct user interviews for applied purposes would be wise to ensure the possibility to pursue research questions concerning potential eHealth working mechanisms by including this purpose in the study information provided to ethics boards and prospective participants. However, getting rich data on eHealth working mechanisms through qualitative interviews may be challenging. In this paper, we suggest that challenges may arise partly due to what we have described as the invisible interaction: that eHealth programs affect the program users’ change processes through their content and how they influence the person-program interaction, but that their influence on the interaction is largely invisible to the user. We have described 5 interview challenges and suggested tools from qualitative methodology to counteract each challenge. These tools may serve as a step toward a set of guidelines for conducting interview studies on eHealth working mechanisms, with the goal of generating rich data that will improve the quality and reach of the findings. Findings from high-quality interview studies can in turn be used to build more general, theoretical knowledge about the working mechanisms of automated eHealth programs. Through theorizing the general working mechanisms of eHealth interventions, we believe that the next generation of eHealth programs can be developed to fully take advantage of this medium’s potential. Interviews with both applied and basic research aims may serve both aims through topical blocks and clear introductions. The transition can be facilitated by structuring the interview in topical blocks [49]: one covering the applied research question, another covering the basic research question. Topical blocks enable the interviewer to focus on 1 research question at a time, facilitating active listening and choosing following-up questions. The topical blocks should be kept separate; therefore, if the participant says something relevant for research question number 1 in the topical block of research question number 2, the interviewer’s follow-up questions on this should be saved for the respective topical block. Furthermore, the transition between the different topical blocks should be made explicit through small introductions: first, a general introduction to the interview along with a presentation of the topical blocks, then separate introductions preceding each topical block. Acknowledgments This research is funded by grant no. 228158/H10 from the Research Council of Norway. The authors wish to thank Filip Drozd, Caroline L Brandt, and Maja Wilhelmsen for contributing to this paper with their experiences with qualitative eHealth research. Their inputs were valuable corrections of the authors’ initial presumptions, and they contributed to making this paper more applicable to a larger part of the field. The authors also wish to thank Professor Azy Barak for his encouragement and valuable insights regarding the need for theoretical work within the field of eHealth. The authors additionally wish to thank Anne-Lise Middelthon for inspiring insights and advice on qualitative methodology. The second author of this paper, Ayna B Johansen, tragically passed away before this paper was published. We are grateful for her enthusiasm for this paper and how she helped shape its content. She is remembered and missed. Structuring the Dual-Aim Interview However, most participants answered interview questions on how they related to the program briefly and superficially, resulting in scant data. The potentially negative influence of the social interview situation can be counteracted with methodological tools. The researcher acknowledging the potential negative influence of roles and stereotypes, both before and after the interviews, can minimize their negative effect. Before an interview, researchers should reflect on potentially salient social aspects and whether something should be done about them [43]. If circumstances Mixing applied and basic research aims can be problematic as different aims may require different interviewing modes. For the interviewer, changing from an applied interviewing mode to a basic interviewing mode will involve changing the point of his or her focal attention, that is, what to listen for and which XSL•FO RenderX XSL•FO RenderX Holter et al JOURNAL OF MEDICAL INTERNET RESEARCH whereas the basic topical block involves a coresearcher design with joint exploration. Providing the interview with structure and appropriate introductions helps both the participant and the interviewer into the right frame of mind, moving from 1 research question to another. In addition, structuring the dual-aim interview into topical blocks ensures that both research questions are being covered, instead of leaving this overview for analysis. follow-up questions to ask. For the participant, changing interviewing modes will involve changing how she or he is expected to answer, from talking more superficially about the breadth of his or her program experiences (applied mode) to talking in depth about a few aspects (basic mode). If the transition between the different modes is not explicit to the participant, she or he may answer interview questions with the wrong mindset —basic interview questions as if they were applied questions or applied interview questions as if they were basic. Unclear transitions may also cause the interviewer to miss important leads in the participant’s answers because of the need to split his or her attention between the 2 research questions. The invisible interaction may exacerbate this challenge: in applied research, the program is treated as a thing, whereas in the search for basic working mechanisms, the program can be considered an interacting agent. An unclear transition between applied and basic research goals may make it more difficult for the participant to take the perspective of the program as an agent influencing the interaction. Concluding Thoughts The introductions can even specify the interviewing modes and what the researcher expects of the participant in each section, for example, that the applied topical block involves factual questions and answers, https://www.jmir.org/2019/5/e10354/ Conflicts of Interest None declared. 1. Hekler EB, Michie S, Pavel M, Rivera DE, Collins LM, Jimison HB, et al. Advancing models and theories for digital behavior change interventions. Am J Prev Med 2016 Nov;51(5):825-832 [FREE Full text] [doi: 10.1016/j.amepre.2016.06.013] [Medline: 27745682] 1. Hekler EB, Michie S, Pavel M, Rivera DE, Collins LM, Jimison HB, et al. Advancing models and theories for digital behavior change interventions. Am J Prev Med 2016 Nov;51(5):825-832 [FREE Full text] [doi: 10.1016/j.amepre.2016.06.013] [Medline: 27745682] 1. Hekler EB, Michie S, Pavel M, Rivera DE, Collins LM, Jimison HB, et al. Advancing models and theories for digital behavior change interventions. Am J Prev Med 2016 Nov;51(5):825-832 [FREE Full text] [doi: 10.1016/j.amepre.2016.06.013] [Medline: 27745682] J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 8 (page number not for citation purposes) J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 8 (page number not for citation purposes) https://www.jmir.org/2019/5/e10354/ XSL•FO RenderX JOURNAL OF MEDICAL INTERNET RESEARCH Holter et al 2. Riley WT, Rivera DE, Atienza AA, Nilsen W, Allison SM, Mermelstein R. Health behavior models in the age of mobile interventions: are our theories up to the task? 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[doi: 10.1177/1354067X16650816] Abbreviations eHealth: electronic health J Med Internet Res 2019 \| vol. 21 \| iss. 5 \| e10354 \| p. 10 (page number not for citation purposes) https://www.jmir.org/2019/5/e10354/ Holter et al JOURNAL OF MEDICAL INTERNET RESEARCH Edited by G Eysenbach; submitted 09.03.18; peer-reviewed by A Lee, A Sheon; comments to author 18.08.18; revise 17.01.19; accepted 07.02.19; published 06.05.19 Please cite as: Holter MTS, Johansen AB, Ness O, Brinkmann S, Høybye MT, Brendryen H Qualitative Interview Studies of Working Mechanisms in Electronic Health: Tools to Enhance Study Quality J Med Internet Res 2019;21(5):e10354 URL: https://www.jmir.org/2019/5/e10354/ doi: 10.2196/10354 PMID: 31066683 ©Marianne TS Holter, Ayna B Johansen, Ottar Ness, Svend Brinkmann, Mette T Høybye, Håvar Brendryen. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 06.05.2019. This is an open-access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work, first published in the Journal of Medical Internet Research, is properly cited. The complete bibliographic information, a link to the original publication on http://www.jmir.org/, as well as this copyright and license information must be included. JOURNAL OF MEDICAL INTERNET RESEARCH https://www.jmir.org/2019/5/e10354/ XSL•FO RenderX	44,113
https://openalex.org/W4312187020	OpenAlex	Open Science	CC-By	2,022	Future Technologies for Train Communication: The Role of LEO HTS Satellites in the Adaptable Communication System	Alessandro Vizzarri	English	Spoken	17,027	28,066	Citation: Vizzarri, A.; Mazzenga, F.; Giuliano, R. Future Technologies for Train Communication: The Role of LEO HTS Satellites in the Adaptable Communication System. Sensors 2023, 23, 68. https://doi.org/10.3390/ s23010068 sensors sensors sensors Alessandro Vizzarri 1,, Franco Mazzenga 2 and Romeo Giuliano 1 Alessandro Vizzarri 1,, Franco Mazzenga 2 and Romeo Giuliano 1 1 Department of Engineering Science, Guglielmo Marconi University, 00198 Rome, Italy 2 Radiolabs Consortium, Department of Enterprise Engineering, University of Roma Tor Vergata, 00133 Rome, Italy y * Correspondence: a.vizzarri@unimarconi.it Abstract: The railway sector has been characterized by important innovations regarding digital tech- nologies for train-to-ground communications. The actual GSM-R system is considered an obsolescent technology expected to be dismissed by 2030. The future communication systems in the rail sectors, such as Adaptable Communication Systems (ACS) and Future Railway Mobile Communication Systems (FRMCS), can manage different bearers as 4G/5G terrestrial technologies and satellites. In this environment, the new High Throughput Satellite (HTS) Low-Earth Orbit (LEO) constellations promise very interesting performances from data rate and coverage points of view. The paper ana- lyzes the LEO constellations of Starlink and OneWeb using public data. The Rome–Florence railway line is considered for simulations. The results evidence the LEO satellite can provide interesting performance in terms of visibility, service connectivity, and trafﬁc capacities (up to 1 Gbps). This feature enables the LEO to fully manage a high amount of data, especially in the railway scenarios of the next years when video data applications will be more present. Keywords: low-earth orbit satellite; 5G; railway; ACS; FRMCS 1. Introduction The European Green Deal deﬁnes important objectives in the transport sector, in particular, climate neutrality by 2050. Involving advanced telecommunication technologies in the rail sector allows not only increasing safety and security of the transport sector but also a reduction in the emissions of CO2. The telecommunication infrastructure is crucial for future train management systems such as European Rail Trafﬁc Management System/European Train Control System (ERTMS/ETCS) in Europe. Academic Editors: Kai Liu, Xinxiang Zhang, Ye Wang and Feng Guo Academic Editors: Kai Liu, Xinxiang Zhang, Ye Wang and Feng Guo y p y p The ERTMS/ETCS are a crucial component of the Adaptable Communication System (ACS) and Future Railway Mobile Communication System (FRMCS), able to use different communication technologies. Received: 28 October 2022 Revised: 12 December 2022 Accepted: 18 December 2022 Published: 21 December 2022 The increase in demand for communication and signaling applications requires broad- band communication systems allowing accurate train positioning, automatic train operation (ATO), and efﬁcient predictive maintenance of rail infrastructure. The 4G/5G terrestrial networks can be affected by: (i) the limitation of the available radio spectrum and (ii) the ne- cessity to identify efﬁcient communication technologies able to overcome the obsolescence of the actual communication standard (GSM-R). The end of life for GSM-R is expected to be around 2030. Citation: Vizzarri, A.; Mazzenga, F.; Giuliano, R. Future Technologies for Train Communication: The Role of LEO HTS Satellites in the Adaptable Communication System. Sensors 2023, 23, 68. https://doi.org/10.3390/ s23010068 2. Review of Satellite Usage in Railway Ref. [3] provides a review of the most important features and impacts introduced by the new Satellite communication (SatCom) technologies, also known as Non-Terrestrial Networks (NTN), according to 3GPP taxonomy. They can provide innovative onboard processing techniques, space data acquisition, collection, and post-analysis. In terms of enabled applications, these new SatCom constellations enable the network integration with terrestrial networks (such as 4G/5G/B5G) and innovative applications in the ﬁeld of earth observation and communications in the transport sectors, such as rail, road, aeronautics, and maritime. Furthermore, the paper underlines that the NTN can improve the 5G network reliability, especially in the case of moving platforms such as cars, trains, and airplanes. The Internet of Things (IoT) networks can also beneﬁt from the NTN. The methodology adopted for the review is based on ﬁve items: system and architecture, air interface, medium access, networking, and test and prototype. Satellite Network Automation, Resource Orchestration, Quantum Key Distribution (QKD) through Optical SatComs, and Machine Learning-based applications are only some key innovative features of NTN. Ref. [4] deﬁnes a techno-economic model to be used for the sustainability assessment of LEO adoption. The LEO constellations provided by Starlink, OneWeb, and Amazon Kuiper are analyzed in terms of Free Space Path Loss, Carrier-to-Noise ratio, and Channel capacity depending on the number of satellites. Finally, the mean capacity per subscriber density is also calculated. The main conclusions underline the role of LEO satellites in communication technologies. The analysis results show that these new SatCom constellations are suitable in the case of a maximum of 0.1 users per km2 if they intend to offer a connectivity service comparable to the other broadband solutions. Ref. [5] is focused on LEO usage in the rail sector, especially in the case of the High- Speed Railway (HSR). The terrestrial networks (such as 4G) can offer a connectivity bitrate up to 100 Mbps covering a cell where HSR is moving up to 350 km/h. The link drop phenomenon and the high number of requested handovers are the main challenges to be managed. LEO systems are a valid alternative to terrestrial communication technologies also thanks to the implementation of content delivery able to guarantee an acceptable delay in the order to 20–30 ms. The authors deﬁne an integrated terrestrial-satellite network (ITSN) as providing high data rates and connectivity without interruptions. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). Satellite technologies are broadband communication systems able to integrate terres- trial networks and even to substitute them when terrestrial is not available. The Geostationary Earth Orbit (GEO) and Medium-Earth Orbit (MEO) satellites are already used in the rail sector also for communication applications from/to the train. However, the considerable values of end-to-end delay limit the usage of GEO and MEO in the rail environment [1,2]. https://www.mdpi.com/journal/sensors Sensors 2023, 23, 68. https://doi.org/10.3390/s23010068 Sensors 2023, 23, 68 2 of 20 Both terrestrial communication systems (4G/5G) and GEO/MEO satellite constella- tions are called Traditional Bearers (TB). Due to the previous factors, it is necessary that additional bearers, called Alternative Bearers (AB), should be identified. Among these ABs, the LEO new constellations arose a crescent interest thanks to the improvement of performance concerning GEO/MEO satellites. The paper is organized as follows. Section 2 presents the state of the art of usage of LEO constellations in the rail sector. Section 3 describes the railway scenarios and applications present in the railway context. Section 4 analyzes the architectures of ACS and FRMCS systems. Section 5 reports the LEO satellites’ role within the FRMCS/ACS infrastructure. Section 6 provides the simulation performed with the LEO constellations (such as Starlink and OneWeb). The corresponding simulation results are also included. Section 7 provides a performance analysis of LEO and GEO satellites. The LEO spectral efﬁciency analysis is also presented. Finally, the main conclusions are drawn in Section 8. 2. Review of Satellite Usage in Railway ITSN can integrate different communication technologies (also known as bearers) thanks to the implementation of the Multipath TCP (MPTCP) protocol. The role of caches is essentially to combine the trafﬁc data and to serve the end-users proactively. In [6], the Intelligent Train Tracking System (ITTS) is analyzed in terms of uplink Medium Access Control (MAC) protocol and the corresponding probability of train de- tection enabled by the usage of LEO satellites. In particular, a Very High Throughput (VHT) Medium Access Control (MAC) protocol is analyzed and deﬁned as a discrete-time Markov Chain. The analysis results show that the VHT MAC protocol provides a maximum Sensors 2023, 23, 68 3 of 20 throughput better than Slotted Aloha (SA) MAC protocol. It makes ITTS more reliable, robust, and resilient to LEO satellites. throughput better than Slotted Aloha (SA) MAC protocol. It makes ITTS more reliable, robust, and resilient to LEO satellites. Ref. [7] presents a review of the 3rd Generation Partnership Project (3GPP). It also underlines the role of LEO SatComs as innovative communication technology supporting the terrestrial 5G networks and beyond. All the satellite vendors need to be involved more and more in the activities of 3GPP and other standardization bodies on 5G technology, especially 5G New Radio (NR). In the case of Sixth Generation (6G) systems, the LEO satellites play a crucial role since they can offer ubiquitous global connectivity. In [8] the role of LEO in future autonomous transportation systems is described in terms of signal latency, cost, and performance. The paper provides a review of the main methodologies to optimize the LEO communication systems, focusing on the space segment, ground segment, user segment, LEO Networks, impact on autonomous vehicles, and cost/coverage issues. The authors also consider the high-speed trains to be served by 5G and LEO systems, together with Machine Learning-based approaches. They underline the important role of LEO communication systems for communication, navigation, and sensing functionalities in the autonomous transport sector. In [9], the Second Generation Digital Video Broadcasting via Satellite (DVB-S2) is used for datacast transmission for railway applications requiring data integrity and guaranteeing the QoS targets. The paper proposes unicast features to support the mobility of devices. The authors deﬁne an innovative architecture based on two link-layer protocols for data encapsulation: Multiple Protocol Encapsulation (MPE) and Generic Stream Encapsulation (GSE). 3.2. Railway Applications and Trafﬁc Classes 2. Review of Satellite Usage in Railway The paper provides a comparison of them and shows how extending the GSE headers to support the LL-FEC identiﬁer descriptors. The ACS Trafﬁc Class IDs have been deﬁned in accordance with the FRMCS require- ments [2,10]. Table 1 shows the ACS Trafﬁc Classes, while Table 2 the FRMCS requirements. 3.1. Railway Scenarios Trafﬁc Class ID Reference FRMCS Application Category Latency Reliability Setup Time 0 ACS control plane FFS (for further study) FFS FFS 1 Voice Low Normal Normal 2 Critical Voice/Critical Video Low Low Immediate 3 Video/critical data (legacy apps)/Non-critical data Normal Low Normal 4 Very Critical data Ultra-Low Ultra-Low Immediate 5 Critical data Low Ultra-Low Immediate 6 Messaging Best Effort Low Normal 7 File transfer Best Effort Normal Normal Table 2. FRMCS Service Requirements. Link Quality FRMCS–Functional Requirement FRMCS–System Requirement Service Attribute Value Impact Latency Low Ultra-Low ≤10 ms Impact on service responsiveness (i.e., mission-critical services) Latency Low Low ≤100 ms Latency Normal Normal ≤500 ms Latency Normal Best Effort >500 ms Packet Loss (%) High Ultra-Low 1–99.9999% Impact on service throughput Packet Loss (%) High Low 1–99.9% Packet Loss (%) Normal Normal 1–99% Table 2. FRMCS Service Requirements. The ﬁrst priority is to avoid train derailment [11] and then to guarantee the security and safety of the train transport system [12]. Signaling application to support transport operation. Signaling information is exchanged between the train and the Control and Command Center as a burst-based message. The packets report the train position (Posi- tion Report, PR) and the corresponding authorization to move across the line (Movement Authority, MA). The complete mechanism is speciﬁed by the European Rail Trafﬁc Manage- ment System/European Train Control System (ERTMS/ETCS). Critical voice application is performed by the train’s personnel to communicate with the other rail nodes and emer- gency entities. Multi-call can be also performed. Critical video applications are used for surveillance purposes. Critical data applications regard the exchange of data on the safety of the train and the considered rail line, such as the communication with the track-side or eventual railway interruption. Non-critical data applications regard the information on train or railway line maintenance and Passenger Information System. 3.1. Railway Scenarios S2R Ju identiﬁed several railway scenarios [2]. The railway scenarios are essentially grouped into ﬁve categories: mainline, urban/metro, regional, freight, station/yard/depot. The mainline line is the line used by different trains and often it connects towns, representing the main route. In the last years, it was enhanced and upgraded to “High- speed Rail”. In this special mainline, the trains are moving at a higher speed than the traditional rail trafﬁc, thanks to dedicated rolling stock and tracks. In High-speed Rail the trains can reach a speed of 200 km/h, even 250 km/h in some cases. This depends on the speciﬁc conditions of the railway track. In the future, a train is expected to reach a speed of up to 500 km/h. In this case, the safety and reliability of railway lines are crucial aspects to be managed carefully. Voice and data services are the main applications. Urban and metro lines are included in Rapid Transit Railways (RTR) operating in urban and suburban areas. Therefore, they are characterized by public transport with high capacity. RTR lines have also the exclusive right of way and are allocated to speciﬁc assets, such as separated tunnels or elevated railways. Regional railway lines operate out of the urban areas and provide transport services to the passenger connecting different towns and cities. There are several stops across the line. Freight rail lines are characterized by low density and higher distances across several regions. They are often formed by single tracks and they are affected by the lack of modern communication technologies. They are still based on old safety systems and manual operations. Finally, the station, yard, and depot are the last railway scenario categories. The station is a railway location where a passenger train can start, stop or end its journey. The yard is constituted of a group of tracks to arrange the trains (such as train-shunting procedure), and other purposes. The train depot is the location where trains are recovered and maintained. The ACS Trafﬁc Class IDs have been deﬁned in accordance with the FRMCS require- ments [2,10]. Table 1 shows the ACS Trafﬁc Classes, while Table 2 the FRMCS requirements. Sensors 2023, 23, 68 4 of 20 Table 1. ACS Trafﬁc Classes. Table 1. ACS Trafﬁc Classes. 4. ACS and FRMCS System Architecture side, the single rail application can interface with the ACS control side and/or ACS user plane, as shown in Figure 1. application side, the single rail application can interface with the ACS control side and/or ACS user plane, as shown in Figure 1. ACS user plane, as shown in Figure 1. Figure 1. Adaptable Communication System (ACS) architecture. Figure 1. Adaptable Communication System (ACS) architecture. Figure 1. Adaptable Communication System (ACS) architecture. Figure 1. Adaptable Communication System (ACS) architecture Figure 1. Adaptable Communication System (ACS) architecture. Figure 1. Adaptable Communication System (ACS) architecture. The ACS is one of the two communication systems for the railway sector. The other one is represented by the Future Railway Mobile Communication System (FRMCS) frame- work. In 2015, the ETSI Technical Committee for Rail Telecommunications (TC RT) started to work on the Next Generation Radio for Rail and to study the Future Railway Mobile Communication System. The main FRMCS technical standard documents are constituted by ETSI TS 103 764 and ETSI TS 103 765, where FRMCS System Architecture and corre- sponding building blocks and functions are specified respectively as shown in Figure 2 The ACS is one of the two communication systems for the railway sector. The other one is represented by the Future Railway Mobile Communication System (FRMCS) framework. In 2015, the ETSI Technical Committee for Rail Telecommunications (TC RT) started to work on the Next Generation Radio for Rail and to study the Future Railway Mobile Communication System. The main FRMCS technical standard documents are constituted by ETSI TS 103 764 and ETSI TS 103 765, where FRMCS System Architecture and corresponding building blocks and functions are speciﬁed, respectively, as shown in Figure 2. The ACS is one of the two communication systems for the railway sector. The other one is represented by the Future Railway Mobile Communication System (FRMCS) frame- work. In 2015, the ETSI Technical Committee for Rail Telecommunications (TC RT) started to work on the Next Generation Radio for Rail and to study the Future Railway Mobile Communication System. The main FRMCS technical standard documents are constituted by ETSI TS 103 764 and ETSI TS 103 765, where FRMCS System Architecture and corre- sponding building blocks and functions are specified, respectively, as shown in Figure 2. p g g p , p y, g Figure 2. Future Railway Mobile Communication System (FRMCS) system architecture. Figure 2. 4. ACS and FRMCS System Architecture Future Railway Mobile Communication System (FRMCS) system architecture. Figure 2. Future Railway Mobile Communication System (FRMCS) system architecture. Figure 2. Future Railway Mobile Communication System (FRMCS) system architecture. Figure 2. Future Railway Mobile Communication System (FRMCS) system architecture. The FRMCS client is built in the ACS directly in terms of ACS on the server network side and the corresponding app. Theoretically, the ACS should not interact with the MNO network, instead, it is an overlay infrastructure on top of the MNO network. The FRMCS, The FRMCS client is built in the ACS directly in terms of ACS on the server network side and the corresponding app. Theoretically, the ACS should not interact with the MNO network, instead, it is an overlay infrastructure on top of the MNO network. The FRMCS, The FRMCS client is built in the ACS directly in terms of ACS on the server network side and the corresponding app. Theoretically, the ACS should not interact with the MNO network, instead, it is an overlay infrastructure on top of the MNO network. The FRMCS, on the other hand, presupposes the direct interaction of the MNO because the FRMCS server interacts with the MNO network according to the service requirements requested by the terminals on board the train. Recently, it seems that ACS also foresees the possibility of having servers (as in the case of FRMCS servers) that interact with the MNO network. Due to this approach, the two systems, ACS and FRMCS, will be identical from an architectural point of view. ACS uses IETF protocols also for signaling (SIP protocol), precisely because it performs an overlay transmission. 4. ACS and FRMCS System Architecture In the post-GSM-R era, two integrated approaches are deﬁned to overcome the dismis- sion of GSM-R systems. Both of them envisage the possibility of using different carriers as train-to-ground communication technologies to provide multiple connectivities. The technical speciﬁcations of ACS are deﬁned in [1,2]. The speciﬁcations deﬁne an overall system architecture able to use different communication technologies independently. This makes ACS a cost-effective and scalable communication system for managing multiple access networks and enabling the bearer’s independence principle. g p p p The ACS also enables (i) efficient QoS control and management, (ii) throughput enhance- ment through carrier aggregation, and (iii) resilience improvement through data redundancy. As shown in Figure 1, the ACS Gateway (ACS GW) is deployed both on the client-side (ACS Onboard GW ACS OGW) and server side (ACS network GW ACS NGW) The ACS also enables (i) efficient QoS control and management, (ii) throughput enhance- ment through carrier aggregation, and (iii) resilience improvement through data redundancy. ( ) Q g , ( ) g p ment through carrier aggregation, and (iii) resilience improvement through data redundancy. As shown in Figure 1, the ACS Gateway (ACS GW) is deployed both on the client-side (ACS Onboard GW, ACS OGW) and server-side (ACS network GW, ACS NGW). g gg g , ( ) p g y As shown in Figure 1, the ACS Gateway (ACS GW) is deployed both on the client-side (ACS Onboard GW, ACS OGW) and server-side (ACS network GW, ACS NGW). From the bearer point of view, in ACS the Control plane and User plane are separated and the ACS GW interfaces with radio communication bearers at the IP level only. Moreover, the ACS GW performs the following actions: (i) setup of Tunnels over the IP bearers, (ii) intelligent control of bearers, and tunnel monitoring. As mentioned, the ACS creates the tunnels and manages the registration, the management of communication sessions, and the control of information and events services for the applications. From the application Sensors 2023, 23, 68 5 of 20 e ACS atio cation h side, the single rail application can interface with the ACS control side and/or ACS user plane, as shown in Figure 1. application side, the single rail application can interface with the ACS control side and/or ACS user plane, as shown in Figure 1. ACS user plane, as shown in Figure 1. 5.1. LEO Satellites and ACS In the simulations, the LEO satellite constellations from Starlink and OneWeb compa- nies are considered. LEO systems are deﬁned as satellites having elliptical/circular orbits ranging from 500 to 2000 km above the Earth’s surface and below the Inner Van Allen Belt. The orbit period can vary from 90 min to a couple of hours. The LEO system’s radius is from 3000 to 4000 km [13]. The launch of the ﬁrst two Starlink prototypes of LEO satellites started in February 2018. In September 2022, Starlink launched nearly 3000 satellites. The second generation of Starlink LEO satellites will enable them to be connected to the smartphone directly [14]. Sensors 2023, 23, 68 6 of 20 bruary Regarding OneWeb LEO systems, the constellation of 648 satellites will be completed by the end of 2022 [15]. Regarding OneWeb LEO systems, the constellation of 648 satellites will be completed by the end of 2022 [15]. 5.2. Adopted Methodology 5.2. Adopted Methodology The scope of the performance analysis is to study and analyze the LEO signal in terms of SINR and related bit rates offered to the train, considering opportune modulation and code scheme (MCS). The adopted methodology is shown in Figure 3. The scope of the performance analysis is to study and analyze the LEO signal in terms of SINR and related bit rates offered to the train, considering opportune modulation and code scheme (MCS). The adopted methodology is shown in Figure 3. Figure 3. The adopted methodology. Figure 3. The adopted methodology. The workflow in Figure 3 is based on the necessity to compose the LEO Satellite sce- nario. It consists of: The workﬂow in Figure 3 is based on the necessity to compose the LEO Satellite scenario. It consists of: • System modeling; • Radio propagation model; • Identiﬁcation of the railway line; • LEO Ground station coordinates; • Starlink LEO satellites coordinate; • Composition of the LEO satellite scenario; • Simulations of the LEO constellation for the evaluation of the radio link budget. • Simulations of the LEO constellation for the evaluation of the radio link budget. The ﬁrst step is to create the LEO satellite scenario to be considered for the simulation and analysis. To compose this scenario system, modeling is necessary to reference architec- ture, satellite terminal, earth ground station, and railway application. Moreover, the railway line allows a deﬁnition of a realistic scenario, as well as the opportune radio propagation model to be used for the radio link budget. The LEO satellite scenario is completed thanks to the possibility to import the coordinates of real existing LEO constellations (such as Starlink Starlink and One Web). The simulations are then performed to calculate the radio link budget and the corresponding SINR. The simulation results provide the SINR trend values of LEOs signals along the considered railway line. In the present paper, the performance analysis is carried out at the system level. p p p p y y In Figure 4 the considered system architectures for non-terrestrial networks (NTN) networks are shown [15]. In Figure 4 the considered system architectures for non-terrestrial networks (NTN) networks are shown [15]. Figure 4a depicts the reference architecture of solutions for New Radio (NR) to support non-terrestrial networks (NTN) with a transparent payload. 5.2. Adopted Methodology 5.2. Adopted Methodology Figure 4a depicts the reference architecture of solutions for New Radio (NR) to support non-terrestrial networks (NTN) with a transparent payload. Sensors 2023, 23, 68 7 of 20 vel. (NTN) (a) (b) Figure 4. Non-terrestrial networks (NTN) with transparent payload. 3GPP-based solutions for NR (a), The considered LEO system architecture (b). Figure 4. Non-terrestrial networks (NTN) with transparent payload. 3GPP-based solutions for NR (a), The considered LEO system architecture (b). (a) (a) (b) (b) Figure 4. Non-terrestrial networks (NTN) with transparent payload. 3GPP-based solutions for NR (a), The considered LEO system architecture (b). Figure 4. Non-terrestrial networks (NTN) with transparent payload. 3GPP-based solutions for NR (a), The considered LEO system architecture (b). Figure 4a depicts the reference architecture of solutions for New Radio (NR) to sup- port non-terrestrial networks (NTN) with a transparent payload. NTN system architecture with the transparent payload is usually constituted by the following elements: NTN system architecture with the transparent payload is usually constituted by the following elements: • A satellite with a transparent payload and several beams. The Field Of View (FOV) of the satellite is depending on the onboard antenna diagram and minimum elevation angle. • A transparent payload for ﬁltering, converting, and amplifying the radio frequency, modulation, demodulation, coding, and decoding. • The User Equipment (UE) is able to receive the satellite signal in the considered service area. • Satellite gateways for the interconnection with a public data network. g y • The Feeder link between the satellite gateway and the satellite. • The service link between the User Equipment (UE) and the satellite (or UAS platform). Figure 4b shows the considered LEO system architecture with the integration with ACS devices in the rail sector. The LEO terminal and the satellite antenna are installed into the On Board Unit (OBU). The LEO terminal and the satellite antenna are installed into the On Board Unit (OBU). They are both present in the ACS OGW. The Rail App is connected to ACS OGW through a L l A N t k (LAN) ti t d b Eth t t i i t l They are both present in the ACS OGW. The Rail App is connected to ACS OGW through a y p pp g Local Area Network (LAN) connection supported by Ethernet transmission protocol. 5.2. Adopted Methodology 5.2. Adopted Methodology ACS OGW is connected to the LEO satellite in orbit through satellite radio access technology depending on the available modulation and Coding scheme (MCS). As evidenced in Figure 4b, the LEO satellite can be connected to the Radio Block Center (RBC) according to three different possible schemes: (1) the RBC is connected to the 5G network and external data network (i.e., Wide Area Network (WAN) context), (2) the RBC is connected to the external data network (i.e., Wide Area Network (WAN) context), and (3) the RBC is connected directly to the ground station (gateway). The RBC is equipped with a network-side ACS NGW, capable of closing the tunnel (stable between ACS OGW and AVS NGW) and then transmitting the information ﬂow to the server-side Rail App. Sensors 2023, 23, 68 8 of 20 8 of 20 As regards the role of satellite nodes on the network, not having further details on the Starlink/OneWeb LEO transmission format, it has been hypothesized that the satellite is transparent and non-regenerative. That is, it is capable of transmitting to the ground station the information ﬂow it receives from the ACS OGW on board the train. From the point of view of access or switching protocols, if the ACS OGW is connected to the 5G network through a direct connection with the gNB (case (1) of Figure 4b), the service link realizes the Uu Access Interface. In the other cases ((cases (2) and (3)) of Figure 4b), the interconnections are based on the IP protocol. g ), p Since the Modulation and Coding Scheme (MCS) is not known in the case of Starlink and OneWeb constellation, the DVB-S2+RCS protocol was considered, as speciﬁed in [16]. The performance evaluations concerned only the radio access network between the satellite and the OBU not in terms of success rate, as in the case of the 5G network [17]. Since the Modulation and Coding Scheme (MCS) is not known in the case of Starlink and OneWeb constellation, the DVB-S2+RCS protocol was considered, as speciﬁed in [16]. The performance evaluations concerned only the radio access network between the satellite and the OBU not in terms of success rate, as in the case of the 5G network [17]. Since the MCS for Starlink and OneWeb is not available, the performance analysis is based on the C/N + I and the corresponding data rate. 5.2. Adopted Methodology 5.2. Adopted Methodology Since the MCS for Starlink and OneWeb is not available, the performance analysis is based on the C/N + I and the corresponding data rate. The main assumptions regard: • The upstream and downstream paths are analyzed separately; • The LEO operator fully manages the satellite-to-ground gateway and guarantees the necessary reliability and system capacity; necessary reliability and system capacity; • The WAN connection between the ground station and the RBC is not analyzed. The radio propagation model of the Digital Video Broadcasting DVB-S2 satellite com- munication system is used to perform the simulation, as in [18]. The analysis also takes into account the attenuation models defined in the recommendation ITU-R P.618-13 [19]. The Rome–Florence Italian railway line is considered to perform the analysis. It can accommodate the mainline, regional, and freight railway lines. The LEO Ground Stations are placed along the railway line connecting the cities of Rome and Florence. All the ground stations or Refer- ence Points (RPs) are referred to through the corresponding geodetic coordinates. RBC server is located in another Italian city, Bologna. In Table 3 the geodetic coordinates of some ground stations are listed. ble 3. Geodetic coordinates of some ground stations along the Rome–Florence railway line. Table 3. Geodetic coordinates of some ground stations along the Rome–Florence railway line. Table 3. Geodetic coordinates of some ground stations along the Rome–Florence railway line. Station Latitude–Longitude [Degree] Roma, Termini 41.90134, 12.50034 Arezzo 43.47592731434826, 11.797549853039794 Firenze, Santa Maria Novella 43.76737, 11.27318 The Starlink and One Web LEO satellite geodetic coordinates are imported from the corresponding available Two Line Element (TLE) ﬁles [20]. Then, the LEO satellite scenario can be composed in terms of LEO satellites, ground stations, and radio propagation model. The simulations are mainly focused on the radio link budget calculations. In this sense, the main satellite parameters are to be calculated. Starting from the azimuth, elevation, and range values of each satellite concerning each one of the selected ground stations or Reference Points (RPs), the following parameters are calculated: • Best and worst case for LEO satellite in terms of visibility. • Propagation delay of the best satellite. The best satellite is the visible satellite with a minimum distance from the reference point for each time interval. • Propagation delay of the worst satellite. The worst satellite is the visible satellite with a maximum distance from the reference point for each time interval. 5.2. Adopted Methodology 5.2. Adopted Methodology • Distribution of visibility intervals of all satellites for each RP. The satellite is visible when its elevation angle is greater or equal to 15◦. • Distribution of the visibility intervals of the best satellite for each RP. • Link budget to calculate the C/(N + I) ratio. Sensors 2023, 23, 68 9 of 20 The procedure used to evaluate the link budget for each RP considers the best, the worst, and the “persistent” satellites. The persistent satellite is deﬁned as the satellite having the highest visibility time interval from the considered RP. When a speciﬁc satellite becomes no longer visible, the modem selects another satellite with its visibility time interval. The link budget is calculated also considering all the time intervals during the day. The procedure is detailed in the following points. p g p Calculations have been carried out considering the real geodetic coordinates of the Starlink/OneWeb LEO satellites and those of the Reference Points (RP). Calculations have been carried out considering the real geodetic coordinates of the Starlink/OneWeb LEO satellites and those of the Reference Points (RP). 1. Set the physical transmission/reception parameters of LEOs and RPs. As in [16], the transmission/ reception parameters used for the DVB-S2 system have been considered for calculations. 2. Calculate the LEO-RP distances by simulating the movement of LEO satellites and assessing the satellites that are visible from each RP at the speciﬁed time instant. g p 3. Given the LEO-RP distance, calculate the corresponding path loss (free space path loss model is considered). 3. Given the LEO-RP distance, calculate the corresponding path loss (free space path loss model is considered). 4. Calculate the corresponding C/(N + I). Receiver noise parameters and interference effects considered in the calculation are the same in [16]. 4. Calculate the corresponding C/(N + I). Receiver noise parameters and interference effects considered in the calculation are the same in [16]. The following Equation (1) describes how to calculate the radio link budget of the considered LEO satellite constellation [21,22]. The formula is in dB. The following Equation (1) describes how to calculate the radio link budget of the considered LEO satellite constellation [21,22]. The formula is in dB. PRX = PTX + GTX −LTX −FSPL −LM + GRX −LRX (1) (1) where: PRX [dBm]: received power PTX [dBm]: transmitter output power GTX [dBi]: transmitter antenna gain LTX [dB]: transmitter loss FSPL [dB]: Free Space Path Loss LM [dB]: additional losses due to fading, body loss, atmospheric loss, etc. GRX [dBi]: receiver antenna gain LRX [dB]: receiver losses The Carrier-to-(Noise + Interference) ratio is given by the following formula: SINR = C/(N + I) (2) (2) where N is the thermal noise power and I is the interference power received. The Equation (2) allows calculating the following parameters: • The ratio of the energy per transmitted symbol to single-sided noise power spectral density, given by the following Equation (3): Es/(N0 + I0) (3) Es/(N0 + I0) (3) where Es is the energy per useful symbol. • The ratio between the energy per information bit and single-sided noise power spectral density is given by the following Equation (4): Eb/(N0 + I0) (4) (4) where Eb = Es −10 × log10. According to [23] the opportune modulation and coding scheme (MCS) are considered considering the target margin (in dB). The available bit rate is the parameter indicating the Sensors 2023, 23, 68 10 of 20 10 of 20 speed of the LEO communication link. It can be calculated from the Spectral efﬁciency (for example in terms of MCS) and the transmission bandwidth, as speciﬁed in the Equation (5): ulation Results speed of the LEO communication link. It can be calculated from the Spectral efﬁciency (for example in terms of MCS) and the transmission bandwidth, as speciﬁed in the Equation (5): ulation Results Available Bit Rate = Spectral Efﬁciency × bandwidth (5) ults refer to the LEO system model defined in Section 5.1. The rail- orence is considered in the simulations. The RPs are placed along (5) rail- ong Available Bit Rate = Spectral Efﬁciency × bandwidth (5) ults refer to the LEO system model defined in Section 5.1. The rail- orence is considered in the simulations The RPs are placed along 6. Simulation Results ne from Rome Florenc ntioned railway line. The simulation results refer to the LEO system model deﬁned in Section 5.1. The railway line from Rome–Florence is considered in the simulations. The RPs are placed along the mentioned railway line. io e ai ay i e gure 5 shows the number of LEO satellites (represented by the asteriks) from Starlink eWeb that are visible in Italy during the day, i.e., with an elevation angle greater ° As from Figure 5 the number of Starlink satellites is almost constant during the Figure 5 shows the number of LEO satellites (represented by the asteriks) from Starlink and OneWeb that are visible in Italy during the day, i.e., with an elevation angle greater than 15◦. As from Figure 5, the number of Starlink satellites is almost constant during the day, except for some peaks. The number of OneWeb satellites is more variable during the day. As mentioned, it depends on the different deployment strategies of the two companies. . As from Figure 5, the number of Starlink satellites is almost constant during the cept for some peaks. The number of OneWeb satellites is more variable during the s mentioned, it depends on the different deployment strategies of the two compa- (a) (b) Figure 5. The number of LEO Satellites visible during the day: Starlink (a), OneWeb (b). 0 5 10 15 20 25 Time of the Day [h] 20 30 40 50 60 70 80 90 No. of visible Satellites on Italy 0 5 10 15 20 25 Time of the Day [h] 2 4 6 8 10 12 14 16 18 No. of visible Satellites on Italy Figure 5. The number of LEO Satellites visible during the day: Starlink (a), OneWeb (b). 022, 22, x FOR PEER REVIEW As from Figure 5, in the Rome–Florence railway line around n. 30–40 of Star (a) 0 5 10 15 20 25 Time of the Day [h] 20 30 40 50 60 70 80 90 No. of visible Satellites on Italy 022, 22, x FOR PEER REVIEW (b) 0 5 10 15 20 25 Time of the Day [h] 2 4 6 8 10 12 14 16 18 No. of visible Satellites on Italy (a) (b) 5. The number of LEO Satellites visible during the day: Starlink (a), OneWeb (b). Figure 5. The number of LEO Satellites visible during the day: Starlink (a), OneWeb (b). 6. Simulation Results ne from Rome Florenc ntioned railway line. As from Figure 5, in the Rome–Florence railway line around n. 30–40 of Sta As from Figure 5, in the Rome–Florence railway line around n. 30–40 of Starlink LEO satellites are visible, n. 8 in the case of OneWeb. The number of visible LEO satellites depends on the different strategies and launch plans adopted by the two companies. satellites are visible, n. 8 in the case of OneWeb. The number of visible LEO satellites de- pends on the different strategies and launch plans adopted by the two companies. Figure 6 shows the distance (in km) of a couple of LEO Satellites visible from an RP (placed in Rome) in a specific time inter al As from Figure 5, in the Rome–Florence railway line around n. 30–40 of Starlink LEO satellites are visible, n. 8 in the case of OneWeb. The number of visible LEO satellites depends on the different strategies and launch plans adopted by the two companies. satellites are visible, n. 8 in the case of OneWeb. The number of visible LEO satellites de- pends on the different strategies and launch plans adopted by the two companies. Figure 6 shows the distance (in km) of a couple of LEO Satellites visible from an RP (placed in Rome) in a specific time inter al p g p p y p Figure 6 shows the distance (in km) of a couple of LEO Satellites visible from an RP (placed in Rome) in a speciﬁc time interval. (placed in Rome) in a specific time interval. p g p p y p Figure 6 shows the distance (in km) of a couple of LEO Satellites visible from an RP (placed in Rome) in a speciﬁc time interval. (placed in Rome) in a specific time interval. (a) (a) (b) Figure 6. Range distance of a couple of LEO Satellites visible during the day from RP located in Rome: Starlink (a), OneWeb (b). Figure 6. Range distance of a couple of LEO Satellites visible during the day from RP located in Rome: Starlink (a), OneWeb (b). (b) (b) (a) Figure 6. Range distance of a couple of LEO Satellites visible during the day from RP located in Rome: Starlink (a), OneWeb (b). Figure 6. Range distance of a couple of LEO Satellites visible during the day from RP located in Rome: Starlink (a), OneWeb (b). 6. Simulation Results ne from Rome Florenc ntioned railway line. at 1500 km in Bologna and 1200 km in Rome. The same OneWeb satellite can be visible at 1900 km in Rome, and at 2150 km in Bologna during the same time interval (from 1:30 and 2:00 a.m.). For example, the same Starlink satellite can be visible at 8 a.m. at 1500 km in Bologna and 1200 km in Rome. The same OneWeb satellite can be visible at 1900 km in Rome, and at 2150 km in Bologna during the same time interval (from 1:30 and 2:00 a.m.). Figure 8 plots the distance range of the best and worst LEO visible satellites during h d f S li k d i li i d i i l f O W b F S li k h d g g g g ( ) Figure 8 plots the distance range of the best and worst LEO visible satellites during the day for Starlink and in a limited time interval for OneWeb. For Starlink, the second-best available satellite is also included. the day for Starlink and in a limited time interval for OneWeb. For Starlink, the second- best available satellite is also included. (a) (a) (b) Figure 8. Range distance of the best and worst LEO Satellite visible during the day: Starlink (a), OneWeb (b). Figure 8. Range distance of the best and worst LEO Satellite visible during the day: Starlink (a), OneWeb (b). (b) (b) (a) Figure 8. Range distance of the best and worst LEO Satellite visible during the day: Starlink (a), OneWeb (b). Figure 8. Range distance of the best and worst LEO Satellite visible during the day: Starlink (a), OneWeb (b). In Figure 8, the distance ranges of the best and worst satellites are indicated in black and red, respectively. In the case of Starlink, the distance of the second-best available sat- ellite is indicated in blue. For Starlink, the best satellite has a minimum distance variable from 300 to 600 km, while the worst one is from 1300 to 1550 km (even 1600 km). The second “best” satellite ranges from 300 to 800 km. For OneWeb, the best satellite is from 500 to 1600 km, while the worst satellite is around 2200 km. I h l b f h d l In Figure 8, the distance ranges of the best and worst satellites are indicated in black and red, respectively. 6. Simulation Results ne from Rome Florenc ntioned railway line. As from Figure 6, the distance ranges from 1200 to 600 km and from 2600 to 2400 km in the case of Starlink and OneWeb, respectively. Fi 7 h th di t f i l LEO t llit i ibl d i th d As from Figure 6, the distance ranges from 1200 to 600 km and from 2600 to 2400 km in the case of Starlink and OneWeb, respectively. As from Figure 6, the distance ranges from 1200 to 600 km and from 2600 to 2400 km in the case of Starlink and OneWeb, respectively. Figure 7 shows the distance range of a single LEO satellite visible during the day As from Figure 6, the distance ranges from 1200 to 600 km and from 2600 to 2400 km in the case of Starlink and OneWeb, respectively. Sensors 2023, 23, 68 11 of 20 2400 km 11 of 20 2400 km Figure 7 shows the distance range of a single LEO satellite visible during the day from two different RPs, one in Rome and one in Bologna, in the case of Starlink and OneWeb constellations. from two different RPs, one in Rome and one in Bologna, in the case of Starlink and One- Web constellations. (a) (a) (b) Figure 7. Range distance of the same LEO Satellite visible during the day from two Ground Stations (Rome and Bologna): Starlink (a), and OneWeb (b). Figure 7. Range distance of the same LEO Satellite visible during the day from two Ground Stations (Rome and Bologna): Starlink (a), and OneWeb (b). (b) (b) (a) Figure 7. Range distance of the same LEO Satellite visible during the day from two Ground Stations (Rome and Bologna): Starlink (a), and OneWeb (b). Figure 7. Range distance of the same LEO Satellite visible during the day from two Ground Stations (Rome and Bologna): Starlink (a), and OneWeb (b). As from Figure 7, due to the limited distance of about 400 km from Rome to Bologna- Florence in terms of LEO systems coverage, the two different RPs see the same satellite. As from Figure 7, due to the limited distance of about 400 km from Rome to Bologna- Florence in terms of LEO systems coverage, the two different RPs see the same satellite. IEW 12 of 21 For example, the same Starlink satellite can be visible at 8 a.m. 6. Simulation Results ne from Rome Florenc ntioned railway line. Figure 10 shows the vis different PRs, in the case of seen in Rome, while the gre visibility duration is around persistent satellite. Consider (a) (b) Figure 9. Cumulative Density Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day: Starlink (a), OneWeb (b). Figure 9. Cumulative Density Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day: Starlink (a), OneWeb (b). 022, 22, x FOR PEER REVIEW 13 of 21 LEO satellite provides a propagation delay of 4.2–5.6 ms, and 4.5–6 ms, respectively, in 50% and 80% of cases. Figure 10 shows the visibility of the best and most persistent satellites seen from two different PRs, in the case of Starlink. The blue/black curves refer to the persistent satellite seen in Rome, while the green and red curves to the best satellite seen in Bologna. The visibility duration is around 1.5 and 4 minutes in 80% and 50% of cases, considering the persistent satellite. Considering the LEO satellite, the visibility duration decreases to val- ues under 30 and 60 s in 80% and 50% of cases. (b) ation delay of 4.2–5.6 ms, and 4.5–6 ms, respecti ty of the best and most persistent satellites seen fr ink. The blue/black curves refer to the persistent s and red curves to the best satellite seen in Bolog and 4 minutes in 80% and 50% of cases, consider the LEO satellite, the visibility duration decreases (a) visibil persis (b) % and visibil Figure 9. Cumulative Density Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day: Starlink (a), OneWeb (b). Figure 9. Cumulative Density Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day: Starlink (a), OneWeb (b). p g y ues under 30 and 60 s in 80% and 50% of cases. 6. Simulation Results ne from Rome Florenc ntioned railway line. In the case of Starlink, the distance of the second-best available satellite is indicated in blue. For Starlink, the best satellite has a minimum distance variable from 300 to 600 km, while the worst one is from 1300 to 1550 km (even 1600 km). The second “best” satellite ranges from 300 to 800 km. For OneWeb, the best satellite is from 500 to 1600 km, while the worst satellite is around 2200 km. Sensors 2023, 23, 68 12 of 20 is from 12 of 20 is from In Figure 9, the Cumulative Distribution Function (CDF) of the propagation delay the case of the best and worst LEO satellite visible during the day. In Figure 10, the Com- plementary CDF (CCDF) of the visibility range of both the best and persistent satellite In Figure 9, the Cumulative Distribution Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day. In Figure 10, the Complementary CDF (CCDF) of the visibility range of both the best and persistent satellite seen by two RPs (one in Rome and one in Bologna). Both plots are referred to as Starlink and One Web. the case of the best and worst LEO satellite visible during the day. In Figure 10, the Com- plementary CDF (CCDF) of the visibility range of both the best and persistent satellite seen by two RPs (one in Rome and one in Bologna). Both plots are referred to as Starlink and One Web. In Figure 9, the Cumulative Distribution Function (CDF) of the propagation delay in the case of the best and worst LEO satellite visible during the day. In Figure 10, the g y g , plementary CDF (CCDF) of the visibility range of both the best and persistent satellite seen by two RPs (one in Rome and one in Bologna) Both plots are referred to as Starlink g y g Complementary CDF (CCDF) of the visibility range of both the best and persistent satellite seen by two RPs (one in Rome and one in Bologna). Both plots are referred to as Starlink and One Web. seen by two RPs (one in Rome and one in Bologna). Both plots are referred to as Starlink and One Web. (a) 22, 22, x FOR PEER REVIEW LEO satellite provides a pro 50% and 80% of cases. 6. Simulation Results ne from Rome Florenc ntioned railway line. As from Figure 9, Star of 1.8–1.5 ms, and 2−5 ms, 0 1 2 3 4 5 6 7 Satellite Visibility Time Interval - Best and Persistent Satellite [min] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CCDF Persistent Satellite - Site: Roma Best Satellite - Site: Roma Persistent Satellite - Site: Bologna Best Satellite - Site: Bologna As from Figure 9, Starlink best and worst LEO satellite provides a propagation delay of 1.8–1.5 ms, and 2−5 ms, respectively, in 50% and 80% of cases. One Web best and worst (a) (b) Figure 10. Complementary Cumulative Distribution Function (CCDF) of the visibility range of the best and most persistent satellite in visibility, seen from two different Ground Stations (Rome and Bologna): Starlink (a), OneWeb (b). 0 1 2 3 4 5 6 7 Satellite Visibility Time Interval - Best and Persistent Satellite [min] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CCDF Persistent Satellite - Site: Roma Best Satellite - Site: Roma Persistent Satellite - Site: Bologna Best Satellite - Site: Bologna Figure 10. Complementary Cumulative Distribution Function (CCDF) of the visibility range of the best and most persistent satellite in visibility, seen from two different Ground Stations (Rome and Bologna): Starlink (a), OneWeb (b). est and worst LEO satellite provides a propagatio ctively, in 50% and 80% of cases. One Web best an (b) (b) (a) Figure 10. Complementary Cumulative Distribution Function (CCDF) of the visibility range of the best and most persistent satellite in visibility, seen from two different Ground Stations (Rome and Bologna): Starlink (a), OneWeb (b). Figure 10. Complementary Cumulative Distribution Function (CCDF) of the visibility range of the best and most persistent satellite in visibility, seen from two different Ground Stations (Rome and Bologna): Starlink (a), OneWeb (b). Considering OneWeb LEO constellations, the visibility duration of the persistent sat- ellite is around 6.8 and 6.5 min in 80% and 50% of cases. It decreases to 1–2 min and 2–3 min in 80% and 50% of cases. Figures 11 and 12 show the Cumulative Distribution Function (CDF) of the achieva- As from Figure 9, Starlink best and worst LEO satellite provides a propagation delay of 1.8–1.5 ms, and 2−5 ms, respectively, in 50% and 80% of cases. 6. Simulation Results ne from Rome Florenc ntioned railway line. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. ble C/(N + I) in the case of best, worst, and persistent satellites. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. C/(N + I) in the case of best, worst, and persistent satellites. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. data collected from all the RPs placed along the considered railway line. (a) (b) Figure 11. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite 8 10 12 14 16 18 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite Figure 11. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). 2022, 22, x FOR PEER REVIEW 14 of 21 (a) 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite 22, 22, x FOR PEER REVIEW (b) 8 10 12 14 16 18 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite (b) (a) W Figure 11. 6. Simulation Results ne from Rome Florenc ntioned railway line. One Web best and worst LEO satellite provides a propagation delay of 4.2–5.6 ms, and 4.5–6 ms, respectively, in 50% and 80% of cases. ble C/(N + I) in the case of best, worst, and persistent satellites. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. 1 Figure 10 shows the visibility of the best and most persistent satellites seen from two different PRs, in the case of Starlink. The blue/black curves refer to the persistent satellite seen in Rome, while the green and red curves to the best satellite seen in Bologna. The visibility duration is around 1.5 and 4 minutes in 80% and 50% of cases, considering the persistent satellite. Considering the LEO satellite, the visibility duration decreases to values under 30 and 60 s in 80% and 50% of cases. est Satellite ersistent Satellite Worst case Satellite 0.7 0.8 0.9 Considering OneWeb LEO constellations, the visibility duration of the persistent satellite is around 6.8 and 6.5 min in 80% and 50% of cases. It decreases to 1–2 min and 2–3 min in 80% and 50% of cases. Sensors 2023, 23, 68 13 of 20 and 2–3 13 of 20 and 2–3 Figures 11 and 12 show the Cumulative Distribution Function (CDF) of the achievable C/(N + I) in the case of best, worst, and persistent satellites. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. Figures 11 and 12 show the Cumulative Distribution Function (CDF) of the achieva- ble C/(N + I) in the case of best, worst, and persistent satellites. In Figure 11 the plot refers to the downlink path, while in Figure 12 to the uplink path. The CDF takes into account the data collected from all the RPs placed along the considered railway line. (a) (b) Figure 11. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). 6. Simulation Results ne from Rome Florenc ntioned railway line. Considering the OneWeb constellation, the best satellite provides a C/(N + I) value of around 13.8 dB and 12.5 dB in 80% and 50% of cases, and 11.5 dB and 10.5 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the persistent satellite provides a value of C/(N + I) around 12.5 dB and 12 dB in 80% and 50% of cases. Considering the OneWeb constellation, the best satellite provides a C/(N + I) value of around 13.8 dB and 12.5 dB in 80% and 50% of cases, and 11.5 dB and 10.5 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the persistent satellite provides a value of C/(N + I) around 12.5 dB and 12 dB in 80% and 50% of cases. ( ) In Figure 12 the uplink path is analyzed. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is con- sidered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. ( ) In Figure 12 the uplink path is analyzed. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is considered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. In Figure 12 the uplink path is analyzed. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is con- sidered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. In Figure 12 the uplink path is analyzed. 6. Simulation Results ne from Rome Florenc ntioned railway line. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. Considering the OneWeb constellation, the best satellite provides a C/(N + I) value of around 13.8 dB and 12.5 dB in 80% and 50% of cases, and 11.5 dB and 10.5 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the persistent satellite provides a value of C/(N + I) around 12.5 dB and 12 dB in 80% and 50% of cases. Considering the OneWeb constellation, the best satellite provides a C/(N + I) value of around 13.8 dB and 12.5 dB in 80% and 50% of cases, and 11.5 dB and 10.5 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the persistent satellite provides a value of C/(N + I) around 12.5 dB and 12 dB in 80% and 50% of cases. 6. Simulation Results ne from Rome Florenc ntioned railway line. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). Figure 11. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). (a) 12 13 14 15 16 17 18 19 20 21 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite (a) (b) Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). 12 13 14 15 16 17 18 19 20 21 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite 6 8 10 12 14 16 18 20 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). (b) 6 8 10 12 14 16 18 20 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite (b) (a) Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. 6. Simulation Results ne from Rome Florenc ntioned railway line. 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite 8 10 12 14 16 18 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (DL) Best Satellite Persistent Satellite Worst case Satellite Figure 11. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in downlink: Starlink (a), OneWeb (b). nsors 2022, 22, x FOR PEER REVIEW 14 of 21 (a) (b) Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and per- sistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite Finally the persistent satellite 12 13 14 15 16 17 18 19 20 21 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite 6 8 10 12 14 16 18 20 C/(N+I) [dB] 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 CDF (UL) Best Satellite Persistent Satellite Worst case Satellite Figure 12. Cumulative Distribution Function (CDF) of the C/(N + I) in case of best, worst, and persistent satellites, visible by all Ground Stations, in uplink: Starlink (a), OneWeb (b). In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite Finally the persistent satellite Figures 11 and 12 show the Cumulative Distribution Function (CDF) of the achievable C/(N + I) in the case of best, worst, and persistent satellites. 6. Simulation Results ne from Rome Florenc ntioned railway line. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. It reaches a value of 15.4 dB and 15.2 dB in 80% and 50% of cases in the case of the worst satellite. Finally, the persistent satellite provides a C/(N + I) value of around 17.7 dB and 16.7 dB in 80% and 50% of cases. In Figure 11 the downlink path is analyzed. Considering the Starlink constellation, in the case of the best satellite, the C/(N + I) value of the best satellite value is around 18 dB and 17.7 dB, respectively in 80% and 50% of cases. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis The LEO constellations from Starlink can provide a path loss ranging from 175 to 180 dB in the case of the worst satellite. This is the situation of the LEO satellites with a higher distance from the considered terrestrial RP. In the best case, the path loss ranges from 164 dB to 173 dB. This is valid for the closest satellite to the considered terrestrial RP. distance from the considered terrestrial RP. In the best case, the path loss ranges from 164 dB to 173 dB. This is valid for the closest satellite to the considered terrestrial RP. The OneWeb LEO constellations can provide a path loss from 178 to 185 dB in the case of the worst satellite In the best case, it ranges from 167 dB to 180 dB higher distance from the considered terrestrial RP. In the best case, the path loss ranges from 164 dB to 173 dB. This is valid for the closest satellite to the considered terrestrial RP. The OneWeb LEO constellations can provide a path loss from 178 to 185 dB in the case of the worst satellite. In the best case, it ranges from 167 dB to 180 dB. dB to 173 dB. This is valid for the closest satellite to the considered terrestrial RP. The OneWeb LEO constellations can provide a path loss from 178 to 185 dB in case of the worst satellite In the best case it ranges from 167 dB to 180 dB The OneWeb LEO constellations can provide a path loss from 178 to 185 dB in the ca of the worst satellite. In the best case, it ranges from 167 dB to 180 dB. case of the worst satellite. In the best case, it ranges from 167 dB to 180 dB. From the service requirements point of view, the rail applications in the future situ- ation (“tomorrow”) will require more bandwidth than the today situation (“today”), as shown in Table 4 [1,2]. From the service requirements point of view, the rail applications in the future situation (“tomorrow”) will require more bandwidth than the today situation (“today”), as shown in Table 4 [1,2]. Table 4. Rail application requirements. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis The performance of LEO constellations can be also analyzed through a comparison with GEO constellations in terms of path loss [24]. Figure 13 gives a graphic represen- tation of the path loss achievable using GEO and LEO constellations by Starlink (a) and OneWeb (b), in the function of elevation degrees. For both of these LEO constellations, the path loss is calculated considering the best and worst LEO satellites. VIEW 15 of 21 (a) (b) Figure 13. Comparison of GEO and LEO constellations in terms of path loss. Starlink (a), OneWeb (b). Figure 13. Comparison of GEO and LEO constellations in terms of path loss. Starlink (a), OneWeb (b). (b) (a) (b) (a) Figure 13. Comparison of GEO and LEO constellations in terms of path loss. Starlink (a), OneWeb (b). Figure 13. Comparison of GEO and LEO constellations in terms of path loss. Starlink (a), OneWeb (b). Figure 13 compares the path loss achievable by using GEO and LEO satel Figure 13 compares the path loss achievable by using GEO and LEO satelli g p p y g The path loss obtained by HEO satellites is in yellow, by the worst LEO in red and the best LEO in blank. g p p y g The path loss obtained by HEO satellites is in yellow, by the worst LEO in red and the best LEO in blank. According to [24], the path loss achievable by a GEO satellite at 2 GHz frequency ranges from 189 to 192 dB for elevation angles ranging from 10 and 90 degrees. The line- of-sight conditions are assumed and the atmospheric losses are responsible for small de- viations of the mentioned values of path loss. According to [24], the path loss achievable by a GEO satellite at 2 GHz frequency ranges from 189 to 192 dB for elevation angles ranging from 10 and 90 degrees. The line-of-sight conditions are assumed and the atmospheric losses are responsible for small deviations of the mentioned values of path loss. p The LEO constellations from Starlink can provide a path loss ranging from 175 to 180 dB in the case of the worst satellite. This is the situation of the LEO satellites with a higher distance from the considered terrestrial RP. In the best case, the path loss ranges from 164 dB to 173 dB. This is valid for the closest satellite to the considered terrestrial RP. 6. Simulation Results ne from Rome Florenc ntioned railway line. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is considered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. In Figure 12 the uplink path is analyzed. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is con- sidered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. In Figure 12 the uplink path is analyzed. Considering the Starlink constellation, the C/(N + I) is around 18.5 dB and 18 dB in 80% and 50% of cases, if the best satellite is considered. The values decrease to 13.5 dB and 13.2 dB in 80% and 50% of cases if the worst satellite is analyzed. Finally, the analysis of the persistent satellite provides a value of C/(N + I) around 17.5 dB and 16 dB in 80% and 50% of cases. Sensors 2023, 23, 68 14 of 20 Considering the OneWeb constellation, the C/(N + I) is around 12.5 dB and 12 dB in 80% and 50% of cases if the best satellite is analyzed. It is around 10 dB and 9.5 dB in 80% and 50% of cases if the worst satellite is considered. Finally, the persistent satellite provides a value of C/(N + I) around 12 dB and 11 dB in 80% and 50% of cases. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis GEO and LEO comparison in terms of one-way delay and single link Bit Rate in Downlink (DL) and Uplink (UL). As in [25,26], GEO satellites (as Inmarsat) provide a worse performance in terms of one-way delay (250 ms) than the LEO satellites (10–15 ms). The LEO satellite can guarantee a single link bit rate of around 100 Mbps and 20 Mbps in DL and UL, respectively. From Tables 4 and 5, it emerges that the LEO satellites can provide a bit rate of 100 and 20 Mbps in DL and UL, respectively. These values are sufﬁcient to support the bit rate required by both video LQ and critical video applications for the rail sector. There are several beneﬁts and opportunities deriving from the usage of LEO HTS satellites. In terms of capacity, at present, the LEO HTS satellites can guarantee a sufﬁcient capacity to absorb tomorrow’s service trafﬁc of a lift number of trains. From the latency point of view, the GEO satellites are not compatible with the new services provided by the ACS classes that are characterized by low transmission latency, such as Automatic Train Operation (ATO) and Autonomous Train Control (ATC). They are the most latency-sensitive rail applications. The LEO HTS satellites can provide a latency of 5–6 ms compatible with the requirements of rail applications mentioned in Table 2. If the control center can be served by one or at most two satellites, LEO HTS satellites are able to satisfy the requirements of low latency services. They can also satisfy the ultra-low latency requirements thanks to a single LEO HTS satellite (one-hop scenario) that can be seen from two different reference points (RPs) (as in the case of the Rome-Florence rail section). It means in this case the single LEO HTS can serve both the rail control center and the ACS on board the train. LEO HTS satellites can also improve the broadband internet connectivity destinated to the onboard rail personnel (Staff). The rail staff can use more bandwidth. Finally, the trackside and wayside devices across the railway lines can also beneﬁt from the usage of LEO HTS. They can improve the evolution of the trackside technologies used for monitoring the railway ecosystem, such as rolling stock, and tracks. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis Rail Application Number of Trains Today Bit Rate DL [Mbps] Today Bit Rate UL [Mbps] Tomorrow Bit Rate DL [Mbps] Tomorrow Bit Rate UL [Mbps] Video LQ @ 500 kbps 12 0.00 0.00 0.00 14.4 Critical video @500 kbps 12 0.00 0.00 0.00 7.2 Table 4. Rail application requirements. Rail Application Number of Trains Today Bit Rate DL [Mbps] Today Bit Rate UL [Mbps] Tomorrow Bit Rate DL [Mbps] Tomorrow Bit Rate UL [Mbps] Video LQ @ 500 kbps 12 0.00 0.00 0.00 14.4 Critical video @ 500 kbps 12 0.00 0.00 0.00 7.2 Table 4. Rail application requirements Table 4. Rail application requirements. In tomorrow’s situation, the video applications for the rail sector will be more present and will require ever more challenging performance. A number of eight on board the train In tomorrow’s situation, the video applications for the rail sector will be more present and will require ever more challenging performance. A number of eight on board the train Sensors 2023, 23, 68 15 of 20 15 of 20 are considered only for UL. It means the transmission analyzed is from the ACS OGW to the ACS NGW. The video format is Video Low Quality (LQ) at 500 kbps. A single camera for video critical application at 500 kbps is also considered for only tomorrow’s situation. In today’s situation, it is not considered. A total number of 12 trains within the Rome–Florence railway line is also estimated. As from Table 4, the Video LQ is not implemented in today’s situation. In tomorrow’s situation, it will be a reality and it will require a bit rate of 14.4 Mbps only for the UL link. The same is for the critical video. It will be only implemented in tomorrow’s situation and it will require a bit rate of 7.2 Mbps. q p The GEO and LEO satellites provide different performances in terms of bit rate and one-way delay. Table 5 summarizes the main features of Geo and LEO satellites in terms of one-way delay and single link Bit Rate in Downlink (DL) and Uplink (UL). Table 5. GEO and LEO comparison in terms of one-way delay and single link Bit Rate in Downlink (DL) and Uplink (UL). Satellite One-Way Delay [ms] Single Link Bit Rate DL [Mbps] Single Link Bit Rate UL [Mbps] GEO 250 1; 4 1; 4 LEO 10; 15 100 20 Table 5. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis They can also enable the interconnection with the trackside sensor network and they can contribute to the optimization of safety in speciﬁc points considered as critical, and, in general, the trafﬁc management in the railway sector. The performance of LEO constellations can be also analyzed in terms of channel utilization efﬁciency and then spectral efﬁciency. The spectral efﬁciency η is given by the ratio in Equation (6): Sensors 2023, 23, 68 16 of 20 16 of 20 η = C B (6) η = C B (6) where C is the Capacity, and B is the Channel bandwidth. where C is the Capacity, and B is the Channel bandwidth. here C is the Capacity, and B is the Channel bandwidth. The relation between the spectral efﬁciency η and the SINR is given by the Shannon formula that in the more general case can be written as: η = A × log2 (1 + SINR G ) (7) (7) where G is the Gap and A is a constant that has been evaluated as follows. As in [27], it has been observed that Starlink adopts OFDM modulation using modulations- coding-schemes (MCS) ranging from QPSK to 64 QAM and coding from 1/3 to 4/5. p g coding-schemes (MCS) ranging from QPSK to 64 QAM and coding from 1/3 to 4/5. g ( ) g g g Given the table of the minimum SINR required for each MCS and then the correspond- ing spectral efﬁciency, we have determined A and G values, as follows: nMCS ∑ n=1 [ηMCS_n −A × log2 (1 + SINR_n G )] 2 (8) (8) where ηMCS_n is the spectral efﬁciency achieved when the n-th MCS is considered and the SINR_n is the corresponding minimum required SINR. where ηMCS_n is the spectral efﬁciency achieved when the n-th MCS is considered and the SINR_n is the corresponding minimum required SINR. The MCS data used in (8) are reported in Table 6 and we have obtained A = 0.8368 and G = 1.4. Table 6. Different MCS values used for LEO performance analysis. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis As expected, Oneweb provides a low spectral efficiency cor- responding to the 16QAM with a code rate of 1/3 up to 1/2. These values will be improved with the increase the number of LEO satellites to be launched in the next years and the use of lower satellite orbits. In Figure 15 the CDF of the best, persistent and worst LEO satellites in the uplink are depicted. The behaviour of LEO satellites from Starlink is shown in Figure 15a, Oneweb LEO satellites in Figure 15b. (a) , 22, x FOR PEER REVIEW As from Figure 14, In from above 3.1 dB which c mat with a code rate of 2/ responding to the 16QAM with the increase the num use of lower satellite orbit In Figure 15 the CDF depicted. The behaviour o (b) nk, the best satellite provides a spectral efficienc onds to a minimum MCS using 16QAM modu xpected, Oneweb provides a low spectral effic code rate of 1/3 up to 1/2. These values will be LEO satellites to be launched in the next year best, persistent and worst LEO satellites in the u satellites from Starlink is shown in Figure 15a (b) worst L nk is s (a) I depic Figure 14. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). Figure 14. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). LEO satellites in Figure 15b. (a) (a) (b) Figure 15. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). Figure 15. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). (b) (b) (a) Figure 15. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). Figure 15. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). Considerations similar to those carried out for spectral efficiency in downlink can be applied in the uplink case. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis In Figure 15 the CDF of the best, persistent and worst LEO satellites in the uplink are depicted. The behaviour of LEO satellites from Starlink is shown in Figure 15a, Oneweb LEO satellites in Figure 15b. (a) (b) Figure 15. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). Considerations similar to those carried out for spectral efficiency in downlink can be Figure 15. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of uplink transmissions: Starlink (a), OneWeb (b). h f h b d ll h l k (a) (b) Figure 14. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). Figure 14. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). 2022, 22, x FOR PEER REVIEW 18 of 21 As from Figure 14, In Starlink, the best satellite provides a spectral efficiency ranging from above 3.1 dB which corresponds to a minimum MCS using 16QAM modulation for- mat with a code rate of 2/3. As expected, Oneweb provides a low spectral efficiency cor- responding to the 16QAM with a code rate of 1/3 up to 1/2. These values will be improved with the increase the number of LEO satellites to be launched in the next years and the use of lower satellite orbits. In Figure 15 the CDF of the best, persistent and worst LEO satellites in the uplink are depicted. The behaviour of LEO satellites from Starlink is shown in Figure 15a, Oneweb LEO satellites in Figure 15b. (a) (b) Figure 14. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). Figure 14. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). 022, 22, x FOR PEER REVIEW 18 of 21 As from Figure 14, In Starlink, the best satellite provides a spectral efficiency ranging from above 3.1 dB which corresponds to a minimum MCS using 16QAM modulation for- mat with a code rate of 2/3. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis 8 C l i In Figure 15 the CDF of the best, persistent and worst LEO satellites in the uplink are depicted. The behaviour of LEO satellites from Starlink is shown in Figure 15a, Oneweb LEO satellites in Figure 15b. 8. Conclusions The railway sector has been characterized by the introduction of different communi- Considerations similar to those carried out for spectral efﬁciency in downlink can be applied in the uplink case. 7. GEO/LEO Comparison and LEO Spectral Efﬁciency Analysis MCS ηMCS Minimum SNR [dB] per MCS QPSK 0.67 1.00 QPSK 1.00 2.00 QPSK 1.33 4.30 QPSK 1.50 5.50 QPSK 1.60 6.20 16QAM 2.00 7.90 16QAM 2.67 11.30 16QAM 3.00 12.20 16QAM 3.20 12.80 64QAM 4.00 15.30 64QAM 4.50 17.50 64QAM 4.80 18.60 In Figures 14 and 15 we plot the CDF of the spectral efﬁciency evaluated in accordance with (7) where A and G are given by solving the minimization of (8). The best, persistent, and worst satellites both in downlink (DL) and uplink (UL) have been considered. The plot has been obtained using (8) and considering the SINR depicted in Figures 11 and 12. g In Figure 14a we plot the CDF of spectral efﬁciency in (7) for the downlink of the best, persistent and worst Starlink LEO satellites are depicted. In Figure 14a we plot the CDF of spectral efﬁciency in (7) for the downlink of the best, persistent and worst Starlink LEO satellites are depicted. As from Figure 14, In Starlink, the best satellite provides a spectral efﬁciency ranging from above 3.1 dB which corresponds to a minimum MCS using 16QAM modulation format with a code rate of 2/3. As expected, Oneweb provides a low spectral efﬁciency corresponding to the 16QAM with a code rate of 1/3 up to 1/2. These values will be improved with the increase the number of LEO satellites to be launched in the next years and the use of lower satellite orbits. 17 of 20 k of the Sensors 2023, 23, 68 (a) (b) Figure 14. Cumulative Density Function (CDF) of spectral efficiency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). Figure 14. Cumulative Density Function (CDF) of spectral efﬁciency for the best, persistent and worst satellites in case of downlink transmissions: Starlink (a), OneWeb (b). Sensors 2022, 22, x FOR PEER REVIEW 18 of 21 As from Figure 14, In Starlink, the best satellite provides a spectral efficiency ranging from above 3.1 dB which corresponds to a minimum MCS using 16QAM modulation for- mat with a code rate of 2/3. As expected, Oneweb provides a low spectral efficiency cor- responding to the 16QAM with a code rate of 1/3 up to 1/2. These values will be improved with the increase the number of LEO satellites to be launched in the next years and the use of lower satellite orbits. a io e o o tions have been 8. Conclusions It can range from 800 Mbps to some Gbps. To calculate the Bit Rate, the DVB-S2 modulation schemes are considered. The product of spectral efﬁciency with the Bandwidth gives the available minimum bit rate. The simulation results are performed considering the Rome–Florence railway line. They evidence the presence of a group of LEO satellites covering Italy and the railway line. The number of LEO satellites depends on the LEO operator strategy, from 30–40 satellites by Starlink to 8 by OneWeb. The minimum distance ranges from 500 km to 1200 for Starlink and OneWeb satellites, respectively. Due to the distance of 400 km between Rome and Bologna (where the RBC is located), the PRs along this railway see the same satellite. In terms of propagation delay, it can vary from 1.8–1.5 ms, and 2–5 ms, respectively, in 50% and 80% of cases considering Starlink’s best and worst LEO satellites. Considering One Web’s best and worst LEO satellites, it can vary from 4.2–5.6 ms, and 4.5–6 ms, respectively, in 50% and 80% of cases. In terms of C/(N + I) value, Starlink provides downlink/uplink values from 18/18.5 dB (best satellite), 15.4/13.5 dB (worst satellite), and 17.7/17.5 dB (persistent satellite). p One Web provides downlink/uplink values from 13.8/12.5 dB (best satellite), 11.5/10 dB (worst satellite), and 12.5/12 dB (persistent satellite). Providing a considerable capacity enables LEO systems to manage a high quantity of data. This is very important in the case of bandwidth-demanding speciﬁc ACS trafﬁc classes. Moreover, this makes LEO technology a competitor of terrestrial communication technologies, such as 4G/5G, where they are not available in a speciﬁc area. Moreover, the analysis shows that the LEO satellite connectivity service is always guaran- teed to the considered RPs. However, the LEO constellations are affected by the typical issues related to the satellite systems, such as geomorphology, tunnels, or other physical obstacles. These issues have to be considered to model an efficient LEO satellite service. The high data rate and the large radio coverage offered by the LEO technologies can also guarantee high levels of connection reliability. This feature is determinant in the case of mission-critical applications. The LEO satellite operator is committed to managing the QoS/QoE aspects of the services. Future analysis needs to take into account the second generation of LEO satellites promising high performance in terms of coverage and data rate offered to the end user. a io e o o tions have been 8. Conclusions p dismissed by 2030, the ACS and FRMCS constitute the future communication technolo- gies in the rail sector. Beyond the terrestrial 4G/5G technologies, new LEO satellite con- stellations play a crucial role. The objective of the present work is to suggest that the use of LEO HTS satellites for railways can constitute valid alternative bearers for the new ACS/FRMCS communication systems used in the railway sector. These intelligently use all the communication technol- The railway sector has been characterized by the introduction of different communica- tion technologies able to control and manage trains. The train-to-ground communications have been based on the terrestrial communication GSM-R. Since it is expected to be dis- missed by 2030, the ACS and FRMCS constitute the future communication technologies in the rail sector. Beyond the terrestrial 4G/5G technologies, new LEO satellite constellations play a crucial role. y y g y ogies available in a certain place and at a certain time. What we have demonstrated in this article is that the new LEO HTS satellites are able, by themselves (and even in extreme situations, where the satellite is the only technology available), to support traffic for today and, above all, for tomorrow’s scenarios. More chal- The objective of the present work is to suggest that the use of LEO HTS satellites for railways can constitute valid alternative bearers for the new ACS/FRMCS communica- tion systems used in the railway sector. These intelligently use all the communication technologies available in a certain place and at a certain time. ), pp y , , lenging railway applications will be increasingly pervasive, including the new ATO/ATC railway automation services. The paper analyzes the performance in terms of available C/(N + I) and the bit rate What we have demonstrated in this article is that the new LEO HTS satellites are able, by themselves (and even in extreme situations, where the satellite is the only technology available), to support trafﬁc for today and, above all, for tomorrow’s scenarios. More chal- Sensors 2023, 23, 68 18 of 20 lenging railway applications will be increasingly pervasive, including the new ATO/ATC railway automation services. y The paper analyzes the performance in terms of available C/(N + I) and the bit rate offered by LEO satellites. Starlink and OneWeb constellations have been analyzed. The minimum elevation angle is 15 degrees. The aggregated trafﬁc capacity depends on the Modulation and Coding Scheme (MCS). a io e o o tions have been 8. Conclusions The handover mechanism and the service continuity have to be evaluated according to the next generation of LEO satellites. Author Contributions: Conceptualization, F.M., R.G. and A.V.; methodology, F.M. and A.V.; software, F.M. and A.V.; validation, R.G. and F.M.; formal analysis, R.G. and F.M.; investigation, A.V.; resources, F.M. and R.G.; data curation, F.M. and A.V.; writing—original draft preparation, A.V. and F.M.; writing—review and editing, R.G. and A.V.; visualization, A.V.; supervision, F.M. and R.G.; project administration, F.M.; funding acquisition, R.G. and F.M. All authors have read and agreed to the published version of the manuscript. Funding: This research was partially developed within the AB4Rail project: www.ab4rail.eu, (accessed on 1 January 2021). Grant Agreement No. 101014517. AB4Rail project is funded by Shift2Rail Joint Undertaking and European Union’s Horizon 2020 research and innovation programmes. Institutional Review Board Statement: Not applicable. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable. Informed Consent Statement: Not applicable. Data Availability Statement: Not Applicable. Conﬂicts of Interest: The authors declare no conﬂict of interest. Conﬂicts of Interest: The authors declare no conﬂict of interest. 19 of 20 19 of 20 Sensors 2023, 23, 68 References 1. Shift2Rail, X2Rail-1 D3.3, Annex Guidelines for Choice of Technology. 2018. Available online: https://projects.shift2rail.org/s2r_ ip_TD_D.aspx?ip=2&td=0ac388f4-f808-484c-9f5e-432447324e36 (accessed on 30 November 2022). p p p ( ) 2. Shift2Rail, X2Rail-1 D3.1 User & System_Requirements_(Telecommunications). 2018. Available online: h 2. Shift2Rail, X2Rail-1 D3.1 User & System_Requirements_(Telecommunications). 2018. Available online: https://projects.shift2rail. org/s2r_ip_TD_D.aspx?ip=2&td=0ac388f4-f808-484c-9f5e-432447324e36 (accessed on 30 November 2022). 2. Shift2Rail, X2Rail 1 D3.1 User & System_Requirements_(Telecommunications). 2018. 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Second Generation Framing Structure, Channel Coding, and Modulation Systems for Broadcasting, Interactive Services, News Gathering, and Other Broadband Satellite Applications. EBU-UER: Grand-Saconnex, Switzerland, 2014. 24. Lin, X.; Rommer, S.; Euler, S.; Yavuz, E.A.; Karlsson, R.S. 5G from Space: An Overview of 3GPP Non-Terrestrial Networks. IEEE Commun. Stand. Mag. 2021, 5, 147–153. [CrossRef] g 25. E-SAT. Inmarsat’s Services. Available online: https://www.e-sat.fr/en/inmarsat (accessed on 8 December 2022). 20 of 20 Sensors 2023, 23, 68 26. Ookla Insights Articles. Starlink Slowed in Q2, Competitors Mounting Challenges. 2022. Available online: https://www.ookla. com/articles/starlink-hughesnet-viasat-performance-q2-2022 (accessed on 8 December 2022). g p q ( ) 27. Pekhterev, S. The Bandwidth of the StarLink Constellation . . . and the Assessment of Its Potential Subscriber Base in the USA. Sat- Magazine, 2021. Available online: http://www.satmagazine.com/story.php?number=1026762698#:~{}:text=The%20bandwidth% 20of%20the%20StarLink%20satellite%20will%20be%2020%20Gbps,64QAM%20(Quadrature%20Amplitude%20Modulation) (accessed on 30 November 2022). Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.	42,048
https://openalex.org/W2060238548	OpenAlex	Open Science	CC-By	2,007	Postpartum Mastitis and Community-acquired Methicillin-resistant<i>Staphylococcus aureus</i>	Pavani Reddy	English	Spoken	2,516	4,582	Pavani Reddy,* Chao Qi,* Teresa Zembower,* Gary A. Noskin,* and Maureen Bolon* This single-center, case-control study documents a relative increase in methicillin resistance among 48 cases of Staphylococcus aureus–associated postpartum mastitis during 1998–2005. Of 21 cases with methicillin resistance, 17 (81%) occurred in 2005. Twenty (95%) isolates con- tained the Staphylococcus cassette chromosome mec type IV gene; this suggests that the increase is due to community-acquired methicillin-resistant S.aureus. Forty-eight cases of S. aureus–associated PPM were identified during the study period; 21 cases were due to MRSA and 27 cases were due to MSSA. A relative increase in MRSA PPM was noted in the later years of the study (Figure 1, p = 0.04). MRSA and MSSA patients did not differ significantly with respect to age, pregnancy his- tory, or symptoms at the time of initial evaluation. In addi- tion, MRSA and MSSA patients did not differ in terms of potential risk factors for infection, such as diabetes, group B β-hemolytic streptococcus colonization, artificial rup- ture of membranes, epidural anesthesia, vaginal lacera- tions, episiotomy, cesarean section, or intrapartum antibiotic use (Table). P ostpartum mastitis (PPM) occurs in as many as one third of breastfeeding women in the United States and leads to breast abscess formation in ≈10% of cases (1,2). Although breast milk cultures are not routine in PPM man- agement, the growth of potentially pathogenic bacteria (such as β-hemolytic streptococci or Staphylococcus aureus) is associated with longer time to recovery and more frequent abscess formation (3). S. aureus is the most common bacterium isolated from such cultures, represent- ing 37%–50% of isolates (4,5). P Ten (48%) MRSA and 11 (41%) MSSA patients required hospitalization. Although these inpatients did not Figure 1. Cases of Staphylococcus aureus–associated postpartum mastitis at a single institution, 1998–2005. Cochrane-Armitage test for linear trend suggests a relative increase in methicillin-resistant cases during the study period; p = 0.04. Reports of methicillin-resistant S. aureus (MRSA) PPM among young, healthy women lacking traditional risk factors for MRSA have emerged in the past few years (6,7). Isolates in these cases of community-acquired infection (CA-MRSA) remain susceptible to multiple non–β-lactam antibiotics and possess distinct molecular features (8). Although risk factors associated with skin and soft tis- sue infections due to CA-MRSA have been described (8,9), characteristics unique to patients with CA-MRSA PPM are unknown. To identify risk factors, complications, and outcomes among patients with CA-MRSA PPM, we conducted a retrospective, case-control study to include all S. DISPATCHES DISPATCHES (SCC) mec type IV gene, which is commonly associated with community-acquired infection. (SCC) mec type IV gene, which is commonly associated with community-acquired infection. Northwestern University Feinberg School of Medicine, Chicago, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA The Study We considered for analysis all patients from Northwestern University’s Prentice Women’s Hospital and affiliated Lynn Sage Comprehensive Breast Center with wound, fluid, drainage, or breast milk cultures positive for S. aureus from January 1998 through December 2005. Case-patients were defined as patients with PPM and a corresponding culture positive for MRSA. Control- patients were defined as patients with PPM and a corre- sponding culture positive for methicillin-susceptible S. aureus (MSSA). Patients who had no evidence of mastitis or who had a history of MRSA were excluded from the study. SCCmec types I–V were identified by a PCR-based multiplex assay; rapid bacterial DNA extraction and PCR amplification were performed as described elsewhere (10). Pavani Reddy,* Chao Qi,* Teresa Zembower,* Gary A. Noskin,* and Maureen Bolon* Pavani Reddy,* Chao Qi,* Teresa Zembower,* Gary A. Noskin,* and Maureen Bolon* aureus–associated cases at a single institution over an 8- year period. MRSA isolates were analyzed by PCR for the presence of the Staphylococcus cassette chromosome Figure 1. Cases of Staphylococcus aureus–associated postpartum mastitis at a single institution, 1998–2005. Cochrane-Armitage test for linear trend suggests a relative increase in methicillin-resistant cases during the study period; p = 0.04. Figure 1. Cases of Staphylococcus aureus–associated postpartum mastitis at a single institution, 1998–2005. Cochrane-Armitage test for linear trend suggests a relative increase in methicillin-resistant cases during the study period; p = 0.04. Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 13, No. 2, February 2007 298 DISPATCHES DISPATCHES Antimicrobial agent use was documented for 18 of 27 MSSA cases; in all 18 cases, isolates were susceptible to the initial antibiotic of choice. Initial regimens included penicillinase-resistant penicillins (10 patients), first-gener- ation cephalosporins (2 patients), macrolides (1 patient), tetracyclines (1 patient), β-lactam/β-lactamase inhibitors (1 patient), vancomycin (1 patient), and clindamycin (2 patients). Duration of therapy for MSSA PPM, document- ed in 12 of 18 cases, was a median of 13.5 days (range 9–27 days). PPM due to CA-MRSA appears to be increasing at our institution. Among 17 MRSA-infected mothers in 2005, delivery dates spanned >9 months without overlap, which suggests that MRSA was independently acquired rather than outbreak-related. In addition, although isolates were not subjected to molecular typing by pulsed-field gel elec- trophoresis, PCR results suggest that 16 (94%) of MRSA isolates in 2005 were community-acquired. The epidemiology of CA-MRSA PPM is poorly understood. Notably, nearly twice as many MRSA-infect- ed than MSSA-infected women were multiparous in this study (57% vs. 33%, respectively). The prevalence of CA- MRSA is increasing among young children, and intrafa- milial transmission of isolates has been documented (11,12). Therefore, mothers with young children may be at increased risk for CA-MRSA PPM. Alternatively, these patients may serve as a reservoir for MRSA in the commu- nity, transmitting this organism to family members. Medical record review of affected patients did not show transmission of S. aureus to infants or other family members. In 1 MRSA patient, a perirectal abscess devel- oped 5 months after the mastitis resolved. Intraoperative cultures of the abscess grew MRSA with identical suscep- tibilities, which suggests persistent colonization; however, typing of the isolates was not performed. Of 21 MRSA isolates available for PCR analysis, 20 possessed SCCmec IV. The remaining isolate contained SCCmec II (Figure 2) and displayed resistance to clin- damycin. In contrast, 95% of isolates with SCC mec IV were clindamycin susceptible. In the current study, women with MRSA were signifi- cantly less likely to receive adequate and timely antimicro- bial drug treatment, but consequences of this difference are unclear. Lee et al. suggest that small CA-MRSA abscesses in children can be managed effectively with incision and drainage alone (13). Indeed, most women in this study underwent incision and drainage or wound aspiration with- out significant differences in outcomes. DISPATCHES Although MSSA patients were more likely to undergo breast abscess incision and drainage than their MRSA counterparts, both methods are considered appropriate surgical interventions (14). Mastitis and Community-acquired MRSA Mastitis and Community-acquired MRSA In 17 of 21 MRSA cases, antibiotic use was document- ed. Twelve patients received antibiotics effective against MRSA, but only 2 received effective coverage at therapy onset (both received clindamycin). Patients initially received a penicillinase-resistant penicillin (10 patients), a first-generation cephalosporin (3 patients), a β-lactam/β- lactamase inhibitor (1 patient), or some combination of the above (6 patients). Median time to effective coverage for MRSA was 5 days (range 0–16 days); adequate antimicro- bial agents included vancomycin (4 patients), trimetho- prim-sulfamethoxazole (1 patient), clindamycin (9 patients), rifampin (2 patients), or some combination of the above (4 patients). Median duration of therapy, document- ed in 8 of 12 effective regimens, was 19 days (range 14–62 days). differ in duration of symptoms before admission, length of stay, or leukocyte count, MRSA patients were more likely to have fever. One patient in each group required readmis- sion for recurrent symptoms (Table). differ in duration of symptoms before admission, length of stay, or leukocyte count, MRSA patients were more likely to have fever. One patient in each group required readmis- sion for recurrent symptoms (Table). Forty-six study patients had an abscess associated with mastitis; most (39 patients) underwent needle aspira- tion. Of these patients, 7 (41%) MRSA and 5 (23%) MSSA patients required repeat aspiration. Notably, 9 MSSA patients underwent incision and drainage a median of 4.5 days after aspiration (range 0–17 days), whereas only 1 MRSA patient required subsequent débridement (1 day later). Reasons for this difference are not clear; however, the more frequent use of serial ultrasound-guided aspira- tion in breast abscess management in recent years (when most MRSA cases occurred) may account for this finding. 299 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 13, No. 2, February 2007 Conclusions To our knowledge, this is the largest case-control study of patients with MRSA-associated PPM. Although S. aureus is the most common etiologic agent of PPM, cases caused by MRSA have rarely been described. Epidemic MRSA cases, linked to the hospital transmission of a com- munity-acquired isolate, have been observed more recently (6). Our study suggests that CA-MRSA is an increasingly common pathogen in spontaneous cases of PPM. Although related cases of infant infection were not found, charts of household contacts were not reviewed in this study; cases of S. aureus transmission to infants or other family members may have been undetected. Several authors have reported mother-to-infant transmission of MRSA through breast milk (15,16). Although decoloniza- tion measures in MRSA-colonized patients have not demonstrated long-term effectiveness (17), the possibility of infant MRSA acquisition may warrant further evalua- tion of such measures in infected, breastfeeding mothers. Figure 2. A) PCR with specific primers for class B mec complex (1.3 kb) and type 2 ccr complex (1.0 kb) identifies isolates containing Staphylococcus cassette chromosome (SCC) mec type IV: lanes 1, 2, and 4-7. B) When control strains are used, PCR identifies SCCmec type II in isolate 3. Lane M, molecular mass marker As with any retrospective case-control study, ours had several limitations. First, the study population is small, which limits the generalizability of the results. Second, patients were added to the study by using results of posi- tive cultures; consequently, cases likely represented more severe and complicated infections in which cultures were necessary after routine therapeutic measures failed. Third, although PPM has been associated with multiple patient factors (i.e., difficulty breastfeeding, tobacco use, and stress), a thorough risk assessment is limited by retrospec- tive study. In addition, medical record review may not indicate certain CA-MRSA risk factors, such as socioeco- nomic status, history of incarceration, or exposure to day care facilities. Finally, although the study results suggest a Figure 2. A) PCR with specific primers for class B mec complex (1.3 kb) and type 2 ccr complex (1.0 kb) identifies isolates containing Staphylococcus cassette chromosome (SCC) mec type IV: lanes 1, 2, and 4-7. B) When control strains are used, PCR identifies SCCmec type II in isolate 3. Lane M, molecular mass marker Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 13, No. 2, February 2007 300 Mastitis and Community-acquired MRSA References 1. Barbosa-Cesnik C, Schwartz K, Foxman B. Lactation mastitis. JAMA. 2003;289:1609–13. 16. Behari P, Englund J, Alcasid G, Garcia-Houchins S, Weber SG. Transmission of methicillin-resistant Staphylococcus aureus to preterm infants through breast milk. Infect Control Hosp Epidemiol. 2004;25:778–80. 2. Foxman B, D’Arcy H, Gillespie B, Bobo JK, Schwartz K. Lactation mastitis: occurrence and medical management among 946 breast- feeding women in the United States. Am J Epidemiol. 2002;155:103–14. 17. Loveday HP, Pellowe CM, Jones SR, Pratt RJ. A systematic review of the evidence for interventions for the prevention and control of methicillin-resistant Staphylococcus aureus (1996-2004): report to the Joint MRSA Working Party (Subgroup A). J Hosp Infect. 2006;63(Suppl 1):S45–70. 3. Osterman KL, Rahm VA. Lactation mastitis: bacterial cultivation of breast milk, symptoms, treatment, and outcome. J Hum Lact. 2000;16:297–302. 4. Marshall BR, Hepper JK, Zirbel CC. Sporadic puerperal mastitis: an infection that need not interrupt lactation. JAMA. 1975;233:1377–9. Address for correspondence: Pavani Reddy, Division of Infectious Diseases, Northwestern University Feinberg School of Medicine, 676 N Saint Clair St, Suite 200, Chicago, IL 60611, USA; email: p- reddy2@md.northwestern.edu 5. Niebyl JR, Spence MR, Parmley TH. Sporadic (non-epidemic) puerperal mastitis. J Reprod Med. 1978;20:97–100. 6. Saiman L, O’Keefe M, Graham PL III, Wu F, Said-Salim B, Kreiswirth B, et al. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women. Clin Infect Dis. 2003;37:1313–9. NEW Acknowledgments 11. Herold BC, Immergluck LC, Maranan MC, Lauderdale DS, Gaskin RE, Boyle-Vavra S, et al. Community-acquired methicillin-resistant Staphylococcus aureus in children with no identified predisposed risk. JAMA. 1998;279:593–8. MSSA and MRSA isolates were identified from the micro- biology database by Mike Malczynski. MRSA strains used as controls for SCCmec typing, including type I (NCTC10442), type II (N315), type III (85/2082), type IV (CA05), and type V (WIS [WBG8318]-JCSC3624), were kindly provided by Dr Teruyo Ito. We also thank Dr Patricia Garcia for her support and guidance. 12. Jones TF, Creech CB, Erwin P, Baird SG, Woron AM, Schaffner W. Family outbreaks of invasive community-associated methicillin- resistant Staphylococcus aureus infection [cited 2006 Dec 26]. Clin Infect Dis [serial online]. 2006 Mar. Available from http://www.journals.uchicago.edu/CID/journal/issues/v42n9/38813 /38813.web.pdf Dr Reddy is currently a fellow in the Division of Infectious Diseases at Northwestern University Feinberg School of Medicine. Her research and clinical interests include surgery- related infections and multidrug-resistant, nosocomial pathogens. 13. Lee MC, Rios AM, Aten MF, Mejias A, Cavuoti D, McCracken GH Jr, et al. Management and outcome of children with skin and soft tissue abscesses caused by community-acquired methicillin-resist- ant Staphylococcus aureus. Pediatr Infect Dis J. 2004;23:123–7. ant Staphylococcus aureus. Pediatr Infect Dis J. 2004;23:1 14. Dener C, Inan A. Breast abscesses in lactating women. World J Surg. 2003;27:130–3. 15. Kawada M, Okuzumi K, Hitomi S, Sugishita C. Transmission of Staphylococcus aureus between healthy, lactating mothers and their infants by breastfeeding. J Hum Lact. 2003;19:411–7. Mastitis and Community-acquired MRSA recent increase in MRSA PPM, an assessment of incidence would require further prospective analysis. 7. Laibl VR, Sheffield JS, Roberts S, McIntire DD, Trevino S, Wendel GD Jr. Clinical presentation of community-acquired methicillin- resistant Staphylococcus aureus in pregnancy. Obstet Gynecol. 2005;106:461–5. In summary, CA-MRSA has emerged as an increas- ingly common pathogen in PPM. Therapy against CA- MRSA should be considered in refractory or severe cases of PPM until wound, drainage, or breast milk cultures can be obtained. Adjunct surgical drainage or aspiration is often warranted in such cases. Additional study is required to determine the utility of routine cultures in postpartum mastitis, the prevalence of CA-MRSA in this emerging problem, and the consequences of CA-MRSA colonization for breastfeeding infants. 8. Naimi TS, LeDell KH, Como-Sabetti K, Borchardt SM, Boxrud DJ, Etienne J, et al. Comparison of community- and health care-associ- ated methicillin-resistant Staphylococcus aureus infection. JAMA. 2003;290:2976–84. 9. Charlebois ED, Perdreau-Remington F, Kreiswirth B, Bangsberg DR, Ciccarone D, Diep BA, et al. Origins of community strains of methicillin-resistant Staphylococcus aureus. Clin Infect Dis. 2004;39:47–54. 10. Zhang K, McClure J, Elsayed S, Louie T, Conly JM. Novel multi- plex PCR assay for characterization and concomitant subtyping of Staphylococcal cassette chromosome mec types I to IV in methi- cillin-resistant Staphylococcus aureus. J Clin Microbiol. 2005;43:5026–33. Job Postings and Conference Announcements For information on how to publish your job and conference announcements in Emerging Infectious Diseases, please contact the CDC Foundation (http://www.cdcfoundation.org), 50 Hurt Plaza, Suite 765, Atlanta, GA 30303, USA. Phone: (404) 653-0790. Fax: (404) 653-0330. Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 13, No. 2, February 2007 301	15,503
https://openalex.org/W2874742762	OpenAlex	Open Science	CC-By	2,018	Perceptions of traditional Chinese medicine for chronic disease care and prevention: a cross-sectional study of Chinese hospital-based health care professionals	Xiaoqing Fan	English	Spoken	8,081	13,534	Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 https://doi.org/10.1186/s12906-018-2273-y Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 https://doi.org/10.1186/s12906-018-2273-y Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 https://doi.org/10.1186/s12906-018-2273-y Open Access © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Perceptions of traditional Chinese medicine for chronic disease care and prevention: a cross-sectional study of Chinese hospital- based health care professionals Perceptions of traditional Chinese medicine for chronic disease care and prevention: a cross-sectional study of Chinese hospital- based health care professionals Xiaoqing Fan1, Fanli Meng1* , Dahui Wang1, Qing Guo2, Zhuoyu Ji1, Lei Yang1 and Atsushi O Xiaoqing Fan1, Fanli Meng1* , Dahui Wang1, Qing Guo2, Zhuoyu Ji1, Lei Yang1 and Atsushi Ogihara3 Abstract Background: In China, demands for disease prevention and health care and the prevalence of chronic non- communicable diseases have increased. TCM and general hospitals are increasingly utilizing TCM strategies for chronic non-communicable disease care and prevention. This study aimed to investigate health care professionals’ (HCPs’) perceptions of TCM for prevention, their TCM knowledge, and their abilities to provide such services in TCM and general hospitals. Methods: This cross-sectional study investigated Chinese medicine hospitals and Chinese medicine departments in general hospitals in five Chinese cities. A self-designed questionnaire used to study 400 HCPs focused on basic demographic data, the demand for and effects of TCM for prevention and treatment, and their perceptions of such service implementation. The data analysis included chi-squared tests and descriptive and multi-factor analyses. Results: The 335 HCP respondents comprised 230 (68.7%) females and 105 (31.3%) males, 75.5% of whom overall had knowledge of TCM preventive and health care services. Respondents older than 40 years (28.6%) had greater knowledge of and satisfaction with TCM for preventive and health care services than younger respondents. Moreover, 97.7% of the older respondents were clearly willing to provide TCM preventive services for chronic diseases, 67.8% of whom indicated that their hospitals already provided TCM for prevention and treatment. According to the chi-squared test results, the TCM service characteristics in hospitals, hospital outlooks regarding TCM and TCM development in hospitals were the primary factors affecting the respondents’ perceptions of TCM for chronic disease care and prevention. The multivariate analysis showed high satisfaction as significantly associated with older providers and those with lengthier work experience, particularly among those who worked in hospitals that provided typical TCM services and had positive attitudes towards TCM. Conclusion: The study HCPs had relatively satisfactory knowledge of and positive attitudes towards TCM for chronic disease care and prevention and would use it in practice. Their perceptions and satisfaction levels correlated closely with the successful application of TCM for preventive care and treatment in hospitals. While the use of TCM for prevention and treatment was well developed in some hospitals, further improvements are warranted. Keywords: Traditional Chinese medicine, Chronic disease care and prevention, Chinese medicine hospitals, Health care professionals, Perception, Circumstance * Correspondence: fanxq1030@163.com 1Medical School of Hangzhou Normal University, No. 16, Xuelin Street, Jianggan District, Hangzhou City 310036, Zhejiang Province, China Full list of author information is available at the end of the article Background preventive care and treatment systems, which have greatly improved their national health [7]. Although conventional health care has advanced worldwide, demonstrated by lower morbidity and mortality rates and improved life quality, complementary and alternative medicine (CAM) is widely used worldwide for preventive care and treat- ment services [8]. Global utilization of CAM is 9.8–76.0%, varying in the US, UK, Australia, and South Africa [5]. CAM services, such as herbal therapy, massage, acupunc- ture and moxibustion, and folk remedies, are similar in many countries [8]. g Over thousands of years of practice, traditional Chinese medicine (TCM) has accumulated many methods, ap- proaches, and beliefs for chronic disease care and preven- tion, which embody the philosophy of ‘Zhi-Wei-Bing’. Traditional Chinese medicine approaches to prevention and health care include emotional adjustment, seasonal health care, dietary care, herbal therapy, acupuncture, massage, detoxification, foot baths, etc. Traditional Chin- ese medicine is one type of traditional medicine that, as a complement and alternative to Western medicine, is widely accepted in China. Compared to Western medi- cine, traditional medicine has unique advantages, such as its safety, lower cost, effectiveness, and convenience, which leads to its high acceptance in an increasing num- ber of countries [1, 2]. Recently, the ‘Zhi-Wei-Bing’ philosophy has attracted the attention of numerous researchers in China and abroad, and many public health investigations are being performed in China. Most investigators are directing greater attention towards TCM preventive and health care services in the community rather than those by health care professionals in Chinese medicine hospitals. Thus far, the Chinese government has instituted nearly 100 regulations to support the development of TCM, par- ticularly for chronic disease care and prevention. In De- cember 2016, China introduced the first law on TCM [3]. Additionally, the Outline of the Strategic Plan for the De- velopment of Traditional Chinese Medicine (2016–2030) and the Chinese Medicine Health Services Development Plan (2015–2020) detailed TCM service processes and standards for prevention and health care and directed hos- pitals and preventive health care institutions to implement the standards. Most regulations emphasized the import- ance of ‘Zhi-Wei-Bing’, which is crucial to realizing the unique role and advantages of TCM [4]. The concept of ‘Zhi-Wei-Bing’ is not only an important part of TCM but also a unique part of traditional Chinese culture. © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 2 of 10 Page 2 of 10 Background The ‘Zhi-Wei-Bing’ philosophy includes disease prevention, treatment, and rehabilitation, which is similar to the con- cept of prevention described by the World Health Organization (WHO) [5]. Prevention and health care practitioners are the most active factors in traditional Chinese medicine hospitals, which are considered the principle facilities providing TCM preventive and health care services [2, 7]. This study aimed to investigate health care professionals’ atti- tudes, general practices, utilization willingness, and sug- gestions regarding the provision of TCM for prevention and health care in hospitals. This study also explored possible factors affecting health care professionals’ per- ceptions of and satisfaction with TCM for preventive and health care services. The findings will ideally provide guidance for medical professionals not only in China but also in other countries who seek to focus attention on chronic diseases and traditional medicine. Data collections Preventive health care professionals practice in two settings in urban China: independent health care workers work for centres for disease control and prevention and rehabilita- tion organizations, whereas in hospitals, preventive health care professionals provide part-time prevention and health care services in addition to disease treatment [6]. We chose health care professionals working in hospitals as the target respondents in this survey, including medical doctors, nurses, medical technicians, and in-hospital pharmacy staff. The reasons for this decision were that first, their work is particularly for ‘preventive purposes’. Second, they account for 63% of all preventive health care professionals. Last, the primary staff using TCM for chronic disease care and pre- vention are typically found in traditional Chinese medicine hospitals and traditional Chinese medicine departments in general hospitals. Though general hospitals mainly use allo- pathic Western medicine, their TCM departments typically use TCM products and technology for disease prevention and treatment, similar to TCM hospitals. A literature review showed that chronic diseases, such as heart disease, stroke, cancer, diabetes and other chronic non-communicable diseases, resulted in a 63% mortality rate worldwide [2]. The prevalence of chronic disease is notably high in China, and continues to rise. The World Bank estimated that the cost of disease-related medical care (e.g., pertaining to cardiovascular diseases, diabetes, chronic obstructive pulmonary disease, etc.) will grow by nearly 50% in 2010–2030 in China [6]. The number of chronic disease patients, whose medical care costs ac- count for 70% of the total disease cost burden in China, will increase to nearly 300 million [6]. Some studies have shown that one-third of human diseases might be avoided through prevention and appropriate disease management [2]. Thus, the prevention of chronic diseases is increas- ingly important. An appropriate prevention and health care system would play a positive role in promoting the development of social economy. Many countries have established national Page 3 of 10 Page 3 of 10 Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 For the investigation, Beijing, Tianjin, Shanghai, Hangzhou, Guangzhou were chosen as research sites because they were the first to pilot the use of TCM for prevention and health care. Their developments of TCM for pre- ventive and health care services were relatively favourable and representative. The five cities are distrib- uted across the north, centre, and south of China, with similar economic levels, policy environments, and health services development. Measurement The original questionnaire was designed based on the results of the literature review, group discussion and underwent several revisions according to feedback from health care professionals in TCM hospitals and general hospitals. This was aimed to ensure the description and wording was intelligible for all participants and improve the validity of the questionnaire. The pre-survey was conducted in Hangzhou, China, using a total of 80 ques- tionnaires (response rate, 97%). Combined with the sta- tistics from the State Administration of Traditional Chinese Medicine, the sample was representative. Based on the reflects from pre-survey, necessary modifications were made. Then a simple random sampling method was used. At the beginning, the random number table method was used to select five TCM hospitals and five general hospitals in the chosen cities. Then, the ques- tionnaires were distributed to the chief hospital adminis- trators by the State Administration of Traditional Chinese Medicine and were completed by health care Statistical analysis h d b The database was established using Epidata 3.0, and the statistical analysis was performed using SPSS 20.0 for Windows. Categorical data were summarized as percent- ages, and continuous data were aggregated as the mean and standard deviation (SD). Initially, the demographic in- formation of the respondents, their attitudes towards TCM, willingness to utilize TCM and TCM practice were analysed in a descriptive manner. Then, a chi-squared test was used to assess any possible relationships between the respondents’ answers (such as their attitudes, willingness and practice in hospitals) and their gender, age, education level, work experience, and practice of TCM for preven- tion and health care in hospitals. The level of statistical significance was defined as P < 0.05. Data collections The research was reviewed and ap- proved by the Ethics Committee of Hangzhou Normal University. professionals who provided TCM preventive and health care services. Finally, 400 health care professionals were surveyed; after eliminating incomplete and unclear ques- tionnaires, the total number of responses was 373 (93.25%). After deleting administrative staff and support staff who were not health care professionals, 335 target re- spondents remained, yielding a response rate of 83.75%. The information we collected encompassed basic demographic data, the perceptions of health care profes- sionals and the current circumstances, and demands for and effects of TCM for prevention and health care in hospitals. Trained investigators (undergraduates and graduate students) collected the questionnaires immedi- ately upon completion and determined whether any in- formation was missing. When information was missing, the investigators contacted the respondents to modify the questionnaires. Trained investigators were also re- cruited to input the questionnaires. A random assess- ment was conducted before the data entry, and if the error rate exceeded 1%, re-entry was required. y The survey was conducted from July to October 2015. One TCM hospital and one TCM department at a general hospital were selected from each of the five cities. The total sample size of 400 was estimated based on 15–20 times the number of questionnaire questions. According to the National Health and Family Planning Commission of the People’s Republic of China (PRC) and State Admin- istration of Traditional Chinese Medicine (National Statis- tical Abstract of Chinese Medicine (2010–2015)), there were 646,000 TCM health care professionals in TCM hos- pitals and general hospitals in 2015. The number of TCM health care professionals differs between TCM hospitals and general hospitals, and approximately 48,000 TCM health care professionals work in TCM hospitals. The number of health care professionals providing TCM pre- ventive and health care services in Chinese medicine hos- pitals is nearly 3 times the number in general hospitals. Therefore, we selected 60 individuals from TCM hospitals and 20 from general hospitals. The health care profes- sionals included practising physicians (i.e., clinical, TCM and public health physicians), occupational assistant phy- sicians, nurses, medical technicians and pharmacy staff. Results Compared with Western medicine, the advantages of Table 1 Demographic characteristics of the health care professionals (n = 335) Variables Frequency (%) Gender Male 105 (31.3) Female 230 (68.7) Age (years) 20–29 135 (40.3) 30–39 104 (31.1) 40–49 52 (15.5) 50–59 34 (10.1) ≥60 10 (3.0) Education Senior high school/secondary school 9 (2.7) College 31 (9.3) Undergraduate 118 (35.2) Masters and above 177 (52.8) Department Internal medicine 120 (35.8) Surgery 19 (5.7) Obstetrics and gynaecology 13 (3.9) Paediatrics 13 (3.9) Chinese medicine 77 (23.0) Other departments 93 (27.8) Position Practising physician (clinical) 66 (19.7) Practising physician (TCM) 168 (50.1) Practising physician (public health) 4 (1.2) Occupational assistant physician 1 (0.3) Nurse 60 (17.9) Medical technician 27 (8.1) Pharmacy staff 9 (2.7) Title Chief physician 20 (6.0) Deputy chief physician 47 (14.0) Intermediate 107 (31.9) Primary and below 161 (48.1) Work experience ≤1 year 65 (19.4) 2–5 years 82 (24.5) 6–10 years 69 (20.6) 11–15 years 41 (12.2) > 15 years 78 (23.3) Table 2 Health care professionals’ perception of TCM preventive and health care services Items N (%) Familiarity Very familiar 56 (16.7) Basic understanding 197 (58.8) Unsure 49 (14.6) Basically no understanding 28 (8.4) Never heard of it 5 (1.5) Advantages Prevention and control of complications 242 (72.2) Few side effects 255 (76.1) Good healing effect 189 (56.4) High patient adherence 165 (49.3) Low cost 130 (38.8) Easy to use 121 (36.1) Willingness to adopt Yes 166 (49.6) Might try 161 (48.1) No 8 (2.4) Suitable method of promotion Educational presentation 255 (76.1) TV & networking 241 (71.9) Newspapers & books 223 (66.6) Community promotion 211 (63.0) Face-to-face discussion 213 (63.6) Family & friends 117 (34.9) Others 27 (8.1) Table 1 Demographic characteristics of the health care professionals (n = 335) Variables Frequency (%) Gender Male 105 (31.3) Female 230 (68.7) Age (years) 20–29 135 (40.3) 30–39 104 (31.1) 40–49 52 (15.5) 50–59 34 (10.1) ≥60 10 (3.0) Education Senior high school/secondary school 9 (2.7) College 31 (9.3) Undergraduate 118 (35.2) Masters and above 177 (52.8) Department Internal medicine 120 (35.8) Surgery 19 (5.7) Obstetrics and gynaecology 13 (3.9) Paediatrics 13 (3.9) Chinese medicine 77 (23.0) Other departments 93 (27.8) Position Practising physician (clinical) 66 (19.7) Practising physician (TCM) 168 (50.1) Practising physician (public health) 4 (1.2) Occupational assistant physician 1 (0.3) Nurse 60 (17.9) Medical technician 27 (8.1) Pharmacy staff 9 (2.7) Title Chief physician 20 (6.0) Deputy chief physician 47 (14.0) Intermediate 107 (31.9) Primary and below 161 (48.1) Work experience ≤1 year 65 (19.4) 2–5 years 82 (24.5) 6–10 years 69 (20.6) 11–15 years 41 (12.2) > 15 years 78 (23.3) When analysing the relationships between the demo- graphic characteristics and the health care professionals’ perceptions, we found that only position significantly impacted the medical staff’s perception (χ2 = 35.540, P < 0.01); moreover, practising TCM physicians had the deepest knowledge of TCM for prevention and health care (data table not shown). Results Demographic characteristics of health care professionals As shown in Table 1, most (230, 68.7%) of the 335 health care professionals surveyed were female. Most respon- dents were 20–39 years old (n = 135; 40.3%), and 28.6% (n = 96) were 40 years of age and older. The education level of the respondents was relatively high; 88.0% of the participants reported having undergraduate or higher edu- cation, whilst those with a college education or below were in the minority (n = 40; 12.0%). Most (n = 234; 69.8%) of the participants were practising physicians (i.e., TCM or clinical physicians). A small proportion (20%) of the respondents were chief or deputy chief physicians. Re- garding the department in which they worked, 35.8% were in the internal medicine department and 23% were in the TCM department. Approximately half of the respondents had more than 6 years of work experience. Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 4 of 10 When analysing the relationships between the demo- graphic characteristics and the health care professionals’ perceptions, we found that only position significantly impacted the medical staff’s perception (χ2 = 35.540, P < 0.01); moreover, practising TCM physicians had the deepest knowledge of TCM for prevention and health care (data table not shown). Health care professionals’ perception of TCM for prevention and health care Table 2 presents among the 335 health care professionals surveyed, the knowledge rate of TCM for prevention and health care was 75.5%, including familiarity with or a basic understanding of TCM preventive and health care services. Concurrently, the knowledge rate of the group over 40 years of age was higher than that of the group younger than 40 years. Health care professionals’ perception of TCM for prevention and health care Table 2 presents among the 335 health care professionals surveyed, the knowledge rate of TCM for prevention and health care was 75.5%, including familiarity with or a basic understanding of TCM preventive and health care services. Concurrently, the knowledge rate of the group over 40 years of age was higher than that of the group younger than 40 years. Compared with Western medicine, the advantages of TCM for prevention and health care were acknowledged. More than 50% of the respondents believed that the Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 5 of 10 biggest advantages of TCM related to the prevention and control of complications, its few side effects and its satis- factory effects on healing. Overall, most (97.7%) health care professionals had used TCM for chronic disease pre- vention, and most respondents in this study believed that educational presentations, the use of electronic media and networking were effective methods to promote TCM for prevention and health care. promote the role of TCM in chronic disease care and pre- vention (74.3%). In contrast, 23.9% of the health care pro- fessionals believed it necessary to improve their attitudes towards health care delivery. Circumstances and effects of TCM for prevention and treatment in hospitals Table 4 shows that 86.3% of the respondents were sup- portive or completely supportive of using TCM methods of prevention and health care to prevent and control chronic diseases, whilst only 0.9% expressed that they were ‘not supportive’ of or ‘against’ such use. Health care professionals’ practice of TCM for prevention and health care in hospitals Table 3 shows that most (83.8%) of the health care pro- fessionals believed that TCM for prevention and health care was necessary for chronic disease prevention and control, whilst merely 3.3% considered it unnecessary. Moreover, 67.8% of the respondents believed that chronic disease care and prevention in their own hospi- tals already exhibited characteristics of TCM, though 23.0% were uncertain. Health care professionals had different views on the current development of TCM for prevention and health care in hospitals: 50.1% believed that it was well-developed, 45.7% agreed that it has some fundamental development but lacks advantages, and only 4.2% believed the current situation was poorly developed. Table 5 shows that 40% of the health care professionals were either satisfied or absolutely satisfied with the ser- vice diversity, charges, medical facilities and awareness of TCM for prevention and treatment provided by their hospitals. Nearly 50% of health care professionals had a neutral attitude towards TCM preventive health care ser- vice projects. Regarding their overall evaluation of TCM prevention and treatment, 50.4% of the health care pro- fessionals had a neutral attitude, whereas 44.2% were ‘satisfied’ or ‘absolutely satisfied’. Health care professionals believed that an effective method to bolster the use of TCM in prevention and health care was to enhance Chinese medicine knowledge through education and training (74.9%) and to vigorously vice projects. Regarding their overall evaluation of TCM prevention and treatment, 50.4% of the health care pro- fessionals had a neutral attitude, whereas 44.2% were ‘satisfied’ or ‘absolutely satisfied’. Analysis of the factors affecting health care professionals’ satisfaction outlook of hospitals towards TCM for prevention and treatment was a positive factor as was whether chronic disease care and prevention services in hospitals had TCM characteristics. Whilst other conditions remained unchanged, the worse the outlook of the hospital towards TCM prevention and treatment, the greater the possibility of low satisfaction (OR = 0.551, P < 0.01). Additionally, the results also showed that in hospitals where chronic disease care and prevention exhibited characteristics of TCM, health care professionals were more likely to be satisfied (OR = 0.247, P < 0.01). As demonstrated by the univariate analysis (Table 6), age and working experience (P < 0.05) were significantly asso- ciated with health care professionals’ overall satisfaction with TCM for prevention and treatment. Health care pro- fessionals 40 years of age and older or those who had more work experience were more likely to be satisfied with TCM for prevention and treatment in hospitals. Health care professionals 60 years of age and older had the highest overall satisfaction. Additionally, satisfactory development of TCM preventive and health care services, positive outlooks of hospitals towards TCM for prevention and treatment, and chronic disease prevention and treat- ment methods incorporating TCM characteristics were significantly associated with high satisfaction. Health care professionals’ practice of TCM for prevention and health care in hospitals Table 3 Health care professionals’ prevention and treatment practices in hospitals Items N (%) Requirements for implementation Completely needed 137 (40.9) Basically needed 142 (42.4) Uncertain 45 (13.4) Basically not needed 9 (2.7) Completely not needed 2 (0.6) Incorporates elements of TCM Yes 227 (67.8) No 31 (9.3) Uncertain 77 (23.0) Aspects for enhancement Bolster TCM training 251 (74.9) Vigorously promote the role of TCM in chronic disease care and prevention 249 (74.3) Enrich the content of TCM services 224 (66.9) Increase the amount of TCM equipment 194 (57.9) Increase TCM prevention and health care personnel 179 (53.4) Lower fees 88 (26.3) Improve service attitudes 80 (23.9) Others 10 (3.0) Table 4 Circumstances and patient satisfaction regarding TCM for prevention and health care in hospitals N (%) Health care professionals’ attitudes towards TCM for prevention and health care Very supportive 139 (41.5) Supportive 150 (44.8) Neutral 43 (12.8) Not supportive 1 (0.3) Against 2 (0.6) Development of TCM for prevention and health care in hospitals Well developed 168 (50.1) Some basic development but lacks advantages 153 (45.7) Poorly developed 14 (4.2) Patients’ satisfaction towards TCM prevention and health care Absolutely satisfied 32 (9.6) Satisfied 174 (51.9) Neutral 120 (35.8) Not satisfied 8 (2.4) Absolutely not satisfied 1 (0.3) Table 3 Health care professionals’ prevention and treatment practices in hospitals Table 3 Health care professionals’ prevention and treatment practices in hospitals Items N (%) Requirements for implementation Completely needed 137 (40.9) Basically needed 142 (42.4) Uncertain 45 (13.4) Basically not needed 9 (2.7) Completely not needed 2 (0.6) Incorporates elements of TCM Yes 227 (67.8) No 31 (9.3) Uncertain 77 (23.0) Aspects for enhancement Bolster TCM training 251 (74.9) Vigorously promote the role of TCM in chronic disease care and prevention 249 (74.3) Enrich the content of TCM services 224 (66.9) Increase the amount of TCM equipment 194 (57.9) Increase TCM prevention and health care personnel 179 (53.4) Lower fees 88 (26.3) Improve service attitudes 80 (23.9) Others 10 (3.0) Table 4 Circumstances and patient satisfaction regarding TCM for prevention and health care in hospitals N (%) Health care professionals’ attitudes towards TCM for prevention and health care Very supportive 139 (41.5) Supportive 150 (44.8) Neutral 43 (12.8) Not supportive 1 (0.3) Against 2 (0.6) Development of TCM for prevention and health care in hospitals Well developed 168 (50.1) Some basic development but lacks advantages 153 (45.7) Poorly developed 14 (4.2) Patients’ satisfaction towards TCM prevention and health care Absolutely satisfied 32 (9.6) Satisfied 174 (51.9) Neutral 120 (35.8) Not satisfied 8 (2.4) Absolutely not satisfied 1 (0.3) Fan et al. means statistically significan Health care professionals’ practice of TCM for prevention and health care in hospitals BMC Complementary and Alternative Medicine (2018) 18:209 Page 6 of 10 Table 5 Health care professionals’ satisfaction levels towards TCM for prevention and health care Items (i.e., concerning TCM preventive health care service projects) Satisfaction level Absolutely not satisfied Not satisfied Neutral Satisfied Absolutely satisfied Are you satisfied with the diversity of services? 5 (1.5%) 29 (8.7%) 162 (48.4%) 117 (34.9%) 22 (6.6%) Are you satisfied with the charges? 6 (1.8%) 27 (8.1%) 175 (52.2%) 109 (32.5%) 18 (5.4%) Are you satisfied with the medical facilities? 5 (1.5%) 34 (10.1%) 180 (53.7%) 99 (29.6%) 17 (5.1%) Are you satisfied with the promotion methods? 7 (2.1%) 25 (7.5%) 157 (46.9%) 126 (37.6%) 20 (6.0%) Are you satisfied with the overall project? 5 (1.5%) 13 (3.9%) 169 (50.4%) 126 (37.6%) 22 (6.6%) Table 5 Health care professionals’ satisfaction levels towards TCM for prevention and health care Analysis of the factors affecting health care professionals’ satisfaction Discussion Though older individuals might become familiar with TCM at an earl- ier age than young individuals, because chronic diseases are more prevalent among older adults, the latter have greater requirements for preventive and therapeutic ser- vices [8]. Some surveys showed that health care profes- sionals’ knowledge of TCM for prevention and health care were primarily obtained from government-issued literature [2]. Thus, internal promotional systems are an important method to improve their knowledge, along with publicizing the concept and philosophy of TCM. In our study, health care professionals acknowledged that educational presentations, the use of television and net- working were more effective methods of disseminating TCM prevention and treatment information, whilst face-to-face discussions and recommendations from family and friends were less effective. These findings indicate that health care professionals are likely to trust official sources of information; therefore, government, social media and professional bodies might exert a posi- tive effect on the further popularization of TCM for pre- vention and treatment. Approximately half of the health care professionals be- lieved that current TCM preventive and health care ser- vices in the hospital had fundamental advantages, such as a favourable political congruence, professional personnel, broad trust among patients and others; however, some dis- advantages remained. Some findings indicated that only 28.75% of practitioners believed that existing preventive and health care services demonstrated TCM characteris- tics, far less than those who believed that the services had no TCM characteristics (59%) [5, 6]. While despite the overall positive attitude towards CAM therapies, Ghan- aian nurses do not perceive themselves to have sufficient knowledge of CAM [30]. To date, the Chinese govern- ment has introduced and implemented numerous policies and regulations throughout the country, particularly re- garding TCM use in community health service centres. TCM for prevention and treatment is acknowledged and used increasingly by Chinese citizens. However, the use of Experts and researchers concluded that health care practitioners were optimistic about CAM; 67% would rec- ommend its use and 63.3% believed that combination therapy using TCM and Western medicine was superior to Western medicine alone [8, 10, 11]. Medical personnel in Africa also had satisfactory experiences with CAM and identified an urgent need to include CAM in the health systems [12, 13]. The health care professionals in this study were positive about applying and constructing a pre- vention and health care system incorporating TCM char- acteristics. Discussion BMC Complementary and Alternative Medicine (2018) 18:209 Page 7 of 10 Table 7 Logistic regression analysis of the factors affecting health care professionals’ overall satisfaction Input variables β SD Wald χ2 P OR (95% CI) Constant −3.494 1.123 9.687 0.002 0.030 Development of TCM prevention and health care services in working hospitals −0.009 0.431 0.000 0.983 0.991 (0.425, 2.307) Attitude of working hospitals towards TCM for prevention and health care −0.597 0.194 9.423 0.002 0.551 (0.376, 0.806) Whether chronic disease care and prevention in working hospitals has TCM characteristics −1.400 0.284 24.317 0.000* 0.247 (0.141, 0.430) P < 0.05 hospitals should implement chronic disease care and pre- vention projects. Substantial data suggest that health care professionals would prefer to choose and incorporate CAM into their health services [14–17]. Perhaps due to the long history of TCM, its role has been widely con- firmed [18]. The philosophy of ‘Zhi-Wei-Bing’ and its characteristics of simplicity, convenience, lower cost, and effectiveness in TCM have been accepted and promoted widely, and TCM treatment and health care services have been widely applied. Furthermore, perhaps the disadvan- tages of Western medicine in the treatment of chronic dis- eases have fuelled health care professionals’ optimism about TCM [19]. According to patients, CAM may not only cure [20] but also may counterbalance the adverse ef- fects of Western medicine [21–23], improve physical and mental health, promote positive medical outcomes and wellness [24–27], etc. TCM may also enhance the com- prehensive outcomes of preventive and treatment efforts and heighten the effectiveness of health interventions and assessments [28]. The conventional HCPs were ready to accept the idea of integrative medicine and the potential role of TCM products and practices in healthcare delivery [29]. However, practitioners’ understanding of CAM must be improved, with specific attention to issues of safety, regulation of evidence-based practice of CAM products and services in Ghana [29]. technologies in the work, the more HCPs can understand the concept of TCM “Zhi-Wei-Bing”, recognize its neces- sity more, and have greater confidence in implementation. Interestingly, older health care professionals (40 years of age and older) had better knowledge of TCM for pre- vention and treatment than younger health care profes- sionals (40 years of age and younger). Discussion This study investigated health care professionals’ percep- tions of the current circumstances surrounding TCM for prevention and treatment in TCM hospitals and TCM de- partments in general hospitals. A total of 75.5% of the re- spondents had knowledge of TCM for chronic disease care and prevention, which approximated the fair knowledge (50–75%) reported by Suganya et al. in India [9]. To some extent, these results reflect the inadequate understanding of TCM for chronic disease care and prevention among health care professionals. At the same time, we can also see that HCPs with a better knowledge of TCM preventive and health care services are more likely to adopt and apply it in daily work. So it is necessary to improve their knowledge through more TCM training and medical practice. Also the more opportunities of applying TCM preventive health care Furthermore, the significant variables in the univariate analysis were used as independent variables, and all pos- sible factors affecting health care professionals’ overall satisfaction were analysed using a step-wise logistic re- gression model. Finally, as Table 7 summarizes, two in- dependent variables were entered into the regression equation: the outlook of hospitals towards TCM for pre- vention and treatment and whether the chronic disease care and prevention services in hospitals had TCM characteristics. According to the factors affecting the overall satisfaction of the respondents in the logistic regression analysis, the Table 6 Univariate analysis of different population characteristics and satisfaction with TCM for prevention and health care Items Absolutely not satisfied (%) Not satisfied (%) Neutral (%) Satisfied (%) Absolutely satisfied (%) χ2 P Age (years) 20–29 1.9 3.1 47.5 38.1 9.4 40.85a 0.004 30–39 1.8 4.5 53.6 31.8 8.2 40–49 0.0 3.8 44.2 46.2 5.8 50–59 0.0 0.0 70.6 23.5 5.9 ≥60 0.0 9.1 36.4 54.5 0.0 Work experience (years) ≤1 1.4 5.2 42.9 42.9 7.5 28.607a 0.027* 2–5 1.1 1.8 57.7 25.5 13.9 6–10 3.0 7.7 52.4 32.1 4.8 11–15 2.7 4.4 35.4 54.9 2.7 > 15 0.0 1.3 55.1 36.8 6.8 *means statistically significan Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 7 of 10 Fan et al. Discussion The government must vigorously cultivate TCM HCPs, not only to increase training opportunities, but also to im- prove training contents, emphasize skills training, and set training content according to their work characteristics and work needs.The main skills of HCPs can include health status identification technology, health state inter- vention technology, Chinese medicine health assessment and physical fitness assessment, common prevention and rehabilitation programs, TCM preventive health care guidelines for high-risk groups of common diseases, and TCM health care technical operating norms. Furthermore, hospitals’ outlooks on the application of TCM for chronic disease care and prevention and whether the services reflect TCM characteristics might contribute to differences in the perspectives and satisfac- tion levels of health care professionals. Overall, we sug- gest the improved development of TCM preventive and health care services, positive outlooks of hospitals towards TCM for prevention and treatment, and the incorporation of TCM-based methods into hospitals’ practices concerning chronic disease care and preven- tion. Hospitals should strengthen the training of high-level TCM HCPs, improve the reward mechanism, and prevent the outflow of talents. As hospitals become more proactive regarding TCM practice for prevention and treatment, health care professionals will be more likely to have better knowledge of and higher satisfaction with TCM. Traditional Chinese medicine preventive and health care services should be rooted in TCM hospitals and general hospitals as centres of care, and aligned with community health service centres. A training system for HCPs in Chinese medicine should be established. It is necessary to require practitioners to have corresponding qualifications, such as qualifications of practicing physicians. Then a reasonable training plan should be established and training materials should be prepared. And combining job training with college educa- tion will make training more effectively. This study has some limitations. The first is related to the background setting and small sample size. When the questionnaires were distributed, some administrative and support staff surveys were admixed into the survey, which led to smaller matched samples. The five selected cities were TCM pilot locations, and their TCM develop- ment was relatively advanced; thus, the research results were not representative of the whole country. Second, the data were gathered from health care professionals’ self-reporting regarding their TCM awareness and prac- tices, without spot visits and objective verification. Discussion Most respondents were willing to utilize TCM measures for chronic disease care and prevention. This finding correlates with the results of another study in China, in which 86% of the medical personnel had positive attitudes towards TCM prevention and treatment [5]. Furthermore, most health care professionals agreed that Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 8 of 10 TCM for prevention and treatment should be made available in hospitals for both health care professionals and patients. In order to implement evidenced-based practice and teach in line with current evidence nurses need to critically examine and reflect on the impact of culture, society and the media on their own health be- liefs [35]. Regarding the lack of work experience, orga- nized education and training opportunities may address this challenge. The chi-squared test showed a significant association between older age and longer work experi- ence with higher satisfaction with TCM among health care professionals. The enthusiasm of health care profes- sionals directly affects their service attitude and quality; thus, the overall enhancement of TCM for prevention and treatment in hospitals is crucial. conventional methods such as those of Western medicine remains the first and most common choice because of their excellent medical systems and rapid results [12, 31]. Fewer than half of the health care professionals were con- tent with the current TCM preventive and health care ser- vices provided, and half reported having a neutral attitude. This finding indicates that the role of TCM in current pre- vention and health care remains unclear and that the role of TCM has not been fully explored. The characteristics and advantages of TCM for preven- tion and treatment are powerful weapons for chronic dis- ease care and prevention [32]. TCM hospitals and community health service institutions already have the capability to provide TCM prevention and health care ser- vices. However, they still need to diligently study and de- velop TCM prevention and health care service products, and to provide team building for medical personnel. Discussion To the best of our knowledge, this study was one of the few domestic projects to evaluate current TCM for chronic disease care and prevention from the perspective of health care professionals in hospitals rather than com- munities, which might be a starting point for similar follow-up studies. The evaluation of the specific satisfac- tion and outcome indicators of TCM for prevention and treatment is necessary. According to the single factor analysis, position and work experience were significantly associated with health care professionals’ perceptions of TCM for prevention and health care services. Studies in Iran found that 88.4% of the participants had no previous complemen- tary and traditional medicine education [17]. Therefore, the training of health care team members, particularly that of nurses and doctors, regarding the applications, benefits and side effects of complementary and trad- itional medicine is recommended [13]. To address the challenge of inadequate familiarity with and knowledge of TCM in China, continuing education in TCM is rec- ommended for above-mentioned targeted health care professionals [33, 34]. Education for HCPs make us ac- knowledge HCPs’ beliefs strongly influence health teach- ing for patients and families. HCPs need to critically examine and reflect on the impact of culture, society and the media on their own health beliefs [35]. Add- itionally, reliable and accessible information concerning Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Page 9 of 10 Page 9 of 10 Authors’ contributions FXQ was the author of the first draft of the article, was responsible for collecting and processing the original data and was primarily responsible for the research results and authenticity of the data. MFL, the general manager of the study, managed the project funding and the design, writing and editing of the manuscript. MFL assisted with the articles and research materials. JZY performed the data analysis. WDH, GQ, YL and OA assisted with the study design. FXQ is the first author, and MFL is the corresponding author for this manuscript. All authors read and approved the final manuscript. 11. Joubert A, Kidd-Taylor A, Christopher G, Nanda J, Warren R, Lindong I, et al. Complementary and alternative medical practice: self-care preferred vs. practitioner-based care among patients with asthma. J Natl Med Assoc. 2010;102:562–9. 12. Adib-Hajbaghery M, Hoseinian M. Knowledge, attitude and practice toward complementary and traditional medicine among Kashan health care staff, 2012. Complement Ther Med. 2014;22:126–32. 13. Mountifield R, Andrews JM, Mikocka-Walus A, Bampton P. Doctor communication quality and friends’ attitudes influence complementary medicine use in inflammatory bowel disease. World J Gastroenterol. 2015; 21:3663–70. Abbreviations Abbreviations CAM: Complementary and Alternative Medicine; HCP: Health care professional; TCM: Traditional Chinese Medicine; WHO: World Health Organization 5. Hua C, Conghua J. Preventing health care practitioners from preventive service for Chinese medicine investigation and analysis of current situation cognition. J Trad Chin Med Manag. 2009;17:906–9. 6. Zhifang Z, Qing G. Research progress on the integration of disease and health management in traditional Chinese medicine. J Zhejiang Trad Chin Med. 2013;48:386–7. Competing interests 18. Zhou XD, Li L, Hesketh T. Health system reform in rural China: voices of health workers and service-users. Soc Sci Med. 2014;117:134–41. 19. Cai Y, Boyd DL, Coeytaux RR, Østbye T, Wu B, Mao Z. Treatment of chronic conditions with traditional Chinese medicine: findings from traditional Chinese medicine hospitals in Hubei, China. J Altern Complement Med. 2015;21:40–5. Author details 1 1Medical School of Hangzhou Normal University, No. 16, Xuelin Street, Jianggan District, Hangzhou City 310036, Zhejiang Province, China. 2Zhejiang Chinese Medical University, No. 548, Binwen Road, Binjiang District, Hangzhou City 310053, Zhejiang Province, China. 3Faculty of Human Sciences, Waseda University, 1-104 Totsukamachi, Shinjuku-ku, Tokyo 169-8050, Japan. In this study, we surveyed hospital-based health care pro- fessionals concerning TCM for prevention and treatment. The respondents had relatively satisfactory knowledge of and positive attitudes towards TCM for prevention and confirmed its contribution to chronic disease care and prevention. Moreover, their perceptions and satisfaction levels were closely related to the application of TCM for prevention and treatment. Currently, the use of TCM for prevention and treatment has been well developed in some hospitals, though further improvement is needed. To enhance the use of TCM for prevention and treatment, professional training of medical personnel must be pro- vided along with enriched service contents and service levels. This study also provides guidance for establishing chronic disease care and prevention service programs. Received: 10 July 2017 Accepted: 27 June 2018 Availability of data and materials Data are available upon request. 10. Song MH, Ung COL, Lee VWY, Hu Y, Zhao J, Li P, et al. Community pharmacists’ perceptions about pharmaceutical service of over-the-counter traditional Chinese medicine: a survey study in Harbin of China. BMC Complement Altern Med. 2017;17:9. Acknowledgements g The authors wish to thank the ten cooperative hospitals for their help with the data collection, and all the study participants. 7. World Health Organization. WHO tools to prevent and control noncommunicable diseases http://www.who.int/topics/noncommunicable_ diseases/en. Accessed 25 July 2017. References 1. Chinese Medicine Act of The People's Republic of China. http://www.npc. gov.cn/npc/xinwen/syxw/2016-12/25/content_2006116.htm. Accessed 25 Dec 2013. 2. Chujie C, Huafeng P. Guangzhou City Community health services utilization actuality and user demand for medical services in the analysis. Chin Gen Practice. 2010;13:1742–4. 2. Chujie C, Huafeng P. Guangzhou City Community health services utilization actuality and user demand for medical services in the analysis. Chin Gen Practice. 2010;13:1742–4. 2. Chujie C, Huafeng P. Guangzhou City Community health services utilization actuality and user demand for medical services in the analysis. Chin Gen Practice. 2010;13:1742–4. 3. State Administion of Traditional Chin Med of the People’s Republic of China. https://bgs.satcm.gov.cn/zhengcewenjian/. Accessed 17 Dec 2017. p p China. https://bgs.satcm.gov.cn/zhengcewenjian/. Accessed 17 Dec 2017. 4. National Health and Family Planning Commission of China. National Basic Public Health Service (Third Edition). Beijing: Government of China;2017(13). p g g g j 4. National Health and Family Planning Commission of China. National Basic Public Health Service (Third Edition). Beijing: Government of China;2017(13). Ethics approval and consent to participate This research was approved by the Ethics Committee of Hangzhou Normal University on 25 March 2015 (reference number 2015–011). Before starting the questionnaire, all participants who ultimately completed the questionnaire were informed of the purpose, content and other details of our study and completed the questionnaire after providing consent. This study did not involve the participants’ personal data, excluding clinical trials or other treatment interventions. All respondent information is confidential, and the participants may remain informed regarding the research progress of this study and may voluntarily decide whether to continue to participate. 14. Xue CCL, Zhang AL, Lin V, Da Costa C, Story DF. Complementary and alternative medicine use in Australia: a national population-based survey. J Altern Complement Med. 2007;13:643–50. 15. Metcalfe A, Williams J, McChesney J, Patten SB, Jetté N. Use of complementary and alternative medicine by those with a chronic disease and the general population – results of a national population based survey. BMC Complement Altern Med. 2010;10:58. 16. Ock SM, Choi JY, Cha YS, Lee J, Chun MS, Huh CH, et al. The use of complementary and alternative medicine in a general population in South Korea: results from a national survey in 2006. J Korean Med Sci. 2009;24:1–6. 17. Eisenberg DM, Davis RB, Ettner SL, Appel S, Wilkey S, Van Rompay M, et al. Trends in alternative medicine use in the United States, 1990-1997: results of a follow-up national survey. JAMA. 1998;280:1569–75. 17. Eisenberg DM, Davis RB, Ettner SL, Appel S, Wilkey S, Van Rompay M, et al. Trends in alternative medicine use in the United States, 1990-1997: results of a follow-up national survey. JAMA. 1998;280:1569–75. 18. Zhou XD, Li L, Hesketh T. Health system reform in rural China: voices of health workers and service-users Soc Sci Med 2014;117:134–41 Competing interests The authors declare that they have no competing interests. Funding Thi 8. Jonas WB. Advising patients on the use of complementary and alternative medicine. Appl Psychophysiol Biofeedback. 2001;26:205–14. This research was supported by funding from the State Administration of Traditional Chinese Medicine of the People’s Republic of China (2012BAI41B00). 9. Suganya M, Vikneshan M, Swathy U. Usage of complementary and alternative medicine: a survey among Indian dental professionals. Complement Ther Clin Practice. 2017;26:26–9. Availability of data and materials Data are available upon request. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Page 10 of 10 Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 20. Sawni A, Thomas R. Pediatricians’ attitudes, experience and referral patterns regarding complementary/alternative medicine: a national survey. BMC Complement Altern Med. 2007;7:18. 21. Posadzki P, Watson LK, Alotaibi A, Ernst E. Prevalence of use of complementary and alternative medicine (CAM) by patients/consumers in the UK: systematic review of surveys. Clin Med. 2013;13:126–31. 22. Sirois FM. Provider-based complementary and alternative medicine use among three chronic illness groups: associations with psychosocial factors and concurrent use of conventional health-care services. Complement Ther Med. 2008;16:73–80. 23. Fletcher CE, Mitchinson AR, Trumble E, Hinshaw DB, Dusek JA. Providers’ and administrators’ perceptions of complementary and integrative health practices across the veterans health administration. J Altern Complement Med. 2017;23:26–34. 24. Birdee GS, Phillips RS, Davis RB, Gardiner P. Factors associated with pediatric use of complementary and alternative medicine. Pediatrics. 2010;125:249–56. 25. Melchart D, Gaisbauer M, Brenke R, Riker U, Liao JZ, Hager S, et al. Observational studies in a network of hospitals using complementary medicine – part I: methods and comparative overview of the results. Forsch Komplementarmed. 1998;5:18–25. 26. Arthur K, Belliard JC, Hardin SB, Knecht K, Chen CS, Montgomery S. Reasons to use and disclose use of complementary medicine use – an insight from cancer patients. Cancer Clin Oncol. 2013;2:81–92. 27. Melchart D, Hager S, Liao JZ, et al. Observational studies in a network of hospitals using complementary medicine. Part II: detailed results from the Hospital for Traditional Chinese Medicine in Kotzting. Forsch Komplementmed. 1998;5:81–6. 28. Nworu CS, Udeogaranya PO, Okafor CK, Adikwu AO, Akah PA. Perception, usage and knowledge of herbal medicines by students and academic staff of University of Nigeria: a survey. Eur J Intergr Med. 2015;7(3):218–27. 29. Kretchy IA, Okere HA, Osafo J, Afrane B, Sarkodi J, Debrah P, et al. Perceptions of traditional, complementary and alternative medicine among conventional healthcare practitioners in Accra, Ghana: implications for integrative healthcare. J Integr Med. 2016;14(5):380–8. 30. Gyasi RM, Abass K, Adu-Gyamfi S, Accam BT. Nurses' knowledge, clinical practice and attitude towards unconventional medicine: implications for intercultural healthcare. Complement Ther Clin Pract. 2017;29:1–8. 31. Bahall M, Legall G. Knowledge, attitudes, and practices among health care providers regarding complementary and alternative medicine in Trinidad and Tobago. BMC Complement Altern Med. 2017;17:144. 32. Chang LH, Wang J. Fan et al. BMC Complementary and Alternative Medicine (2018) 18:209 Publisher’s Note Integration of complementary medical treatments with rehabilitation from the perspectives of patients and their caregivers: a qualitative inquiry. Clin Rehabil. 2009;23:730–40. 33. Clement YN, Williams AF, Khan K, Bernard T, Bhola S, Fortuné M, et al. A gap between acceptance and knowledge of herbal remedies by physicians: the need for educational intervention. BMC Complement Altern Med. 2005;5:20. 34. Van Staden AM, Joubert BAG. Interest in and willingness to use complementary, alternative and traditional medicine among academic and administrative staff in Bloemfontein, South Africa. Afr J Tradit Complement. 2014;11(5):61–6. 35. Cao R, Stone TE, Petrini MA, Turale S. Nurses' perceptions of health beliefs and impact on teaching and practice: a Q-sort study. Int Nurs Rev. 2018; 65(1):131–44.	27,431
https://openalex.org/W4319065526	OpenAlex	Open Science	CC-By-SA	2,022	PERLINDUNGAN DATA PRIBADI PASIEN TERHADAP SERANGAN CYBER CRIME	Arya Wirai Khalifatullah	Indonesian	Spoken	3,379	8,168	Perlindungan Data Pribadi Pasien Terhadap Serangan Cyber Crime Article Info Article history: Received Dec 9, 2022 Revised Dec 20, 2022 Accepted Dec 31, 2022 Kata Kunci: Perlindungan, Data Pribadi, Cyber Crime Keywords: Protection, Personal Data, Cyber Crime Hak adalah kewenangan (badan hukum) yang dimiliki seseorang untuk memperoleh hak yang diperoleh pasien. Hak pasien adalah mendapatkan penjelasan tentang peraturan rumah sakit, pelayanan medis, atau penyakit pasien. Demikian juga rumah sakit juga harus menginformasikan kepada pasien tentang gangguan kesehatan pasien atau apa yang diderita pasien dengan lancar, jelas, dan pasti, tanpa ada gangguan dari rekam medis, maka kami selaku penyelenggara rumah sakit berusaha semaksimal mungkin melindungi data pribadi pasien. atas penderitaan yang dialami di rumah sakit, karena di era sekarang ini sangat berbahaya adanya berita dari luar tentang pembobolan data pasien di media sosial atau ini sangat mempengaruhi identitas pasien dan pihak rumah sakit untuk menyebarkan nama baik. Walaupun Undang-undang Kesehatan tidak secara jelas menyebutkannya, namun pengertian rumah sakit dapat disimpulkan sebagai salah satu bentuk fasilitas pelayanan kesehatan. Sebagaimana dirumuskan dalam Pasal 1 angka 7 bahwa “Fasilitas pelayanan kesehatan adalah alat dan/atau tempat yang digunakan untuk menyelenggarakan upaya pelayanan kesehatan, baik preventif, kuratif, maupun rehabilitatif yang dilakukan oleh pemerintah daerah dan masyarakat. Sanskara Hukum dan HAM Sanskara Hukum dan HAM Sanskara Hukum dan HAM Vol. 01, No. 02, Desember, pp. 47~53 Vol. 01, No. 02, Desember, pp. 47~53 pp Perlindungan Data Pribadi Pasien Terhadap Serangan Cyber Crime Afifah Fitri Apsari1, Anifatun Lutfiyah2, Arya Wirai Khalifatullah3, Erina Nugrahaningtyas4, Ervina Anisya Qoriah5, Gesit Syaifrudin Zukhri6, Muh. Rizal Rosyid Ridho7 Administrasi Rumah Sakit, Fakultas Ilmu Kesehatan, Universitas Duta Bangsa, Surakarta, Indonesia Jl. KH. Samanhudi 93 Sondakan, Laweyan, Surakarta- Phone 0271-712826 affhftrapsr@gmail.com1, anifatun.lutfiyah@gmail.com2, aryawirai1209@gmail.com3, erinanugraha8@gmail.com4, anisyaqoriah@gmail.com5, gesitzuhri3@gmail.com6, ryzelfoxx12@gmail.com7 Article Info ABSTRAK Article history: Received Dec 9, 2022 Revised Dec 20, 2022 Accepted Dec 31, 2022 Hak adalah kewenangan (badan hukum) yang dimiliki seseorang untuk memperoleh hak yang diperoleh pasien. Hak pasien adalah mendapatkan penjelasan tentang peraturan rumah sakit, pelayanan medis, atau penyakit pasien. Demikian juga rumah sakit juga harus menginformasikan kepada pasien tentang gangguan kesehatan pasien atau apa yang diderita pasien dengan lancar, jelas, dan pasti, tanpa ada gangguan dari rekam medis, maka kami selaku penyelenggara rumah sakit berusaha semaksimal mungkin melindungi data pribadi pasien. atas penderitaan yang dialami di rumah sakit, karena di era sekarang ini sangat berbahaya adanya berita dari luar tentang pembobolan data pasien di media sosial atau ini sangat mempengaruhi identitas pasien dan pihak rumah sakit untuk menyebarkan nama baik. Walaupun Undang-undang Kesehatan tidak secara jelas menyebutkannya, namun pengertian rumah sakit dapat disimpulkan sebagai salah satu bentuk fasilitas pelayanan kesehatan. Sebagaimana dirumuskan dalam Pasal 1 angka 7 bahwa “Fasilitas pelayanan kesehatan adalah alat dan/atau tempat yang digunakan untuk menyelenggarakan upaya pelayanan kesehatan, baik preventif, kuratif, maupun rehabilitatif yang dilakukan oleh pemerintah daerah dan masyarakat. ABSTRACT The right is the authority (legal entity) that a person has to obtain the rights obtained by the patient. The patient's right is to receive an explanation of hospital rules, medical services, or the patient's illness. Likewise, the hospital must also inform the patient about the patient's health problems or what the patient is suffering from smoothly, clearly, and definitely, without any distraction from the medical record, so we as the implementation of the hospital try as much as possible to protect the patient's personal data for the suffering Kata Kunci: Perlindungan, Data Pribadi, Cyber Crime Keywords: Protection, Personal Data, Cyber Crime ABSTRACT The right is the authority (legal entity) that a person has to obtain the rights obtained by the patient. The patient's right is to receive an explanation of hospital rules, medical services, or the patient's illness. Likewise, the hospital must also inform the patient about the patient's health problems or what the patient is suffering from smoothly, clearly, and definitely, without any distraction from the medical record, so we as the implementation of the hospital try as much as possible to protect the patient's personal data. for the suffering that has been experienced in the hospital, because in this era it is very dangerous to have news from outside about data breaches of patients on social media or this greatly affects the identity of patients and the hospital for spreading good names. Even though the Health Law does not clearly state it, the definition of a hospital can be concluded as a form of health service facility. As formulated in Article 1 point 7 that "Health service facility is a tool and/or Sanskara Hukum dan HAM (SHH) r 48 r 48 place that is used to carry out health service efforts, both preventive, curative and rehabilitative carried out by the regional government and the community. This is an open access article under the CC BY-SA license. This is an open access article under the CC BY-SA license. Corresponding Author: Name: Arya Wirai Khalifatullah Institution: Universitas Duta Bangsa Email: aryawirai1209@gmail.com Name: Arya Wirai Khalifatullah Institution: Universitas Duta Bangsa Email: aryawirai1209@gmail.com Sanskara Hukum dan HAM (SHH) r 49 r Berdasarkan pengertian di atas dapat disimpulkan bahwa Rumah Sakit merupakan usaha yang mempunyai ciri sendiri dan bertujuan pada pemeriksaan medis dan tindakan medis yang dilakukan terhadap pasien rawat jalan dan pasien inap di Rumah Sakit. Tugas Rumah Sakit rumusan yuridisnya dapat dilihat pada ketentuan Pasal 1 butir 1 Undang-undang Nomor 44 Tahun 2009 tentang Rumah Sakit, seperti disebutkan pada pasal ini, bahwa: “Rumah Sakit adalah institusi pelayanan kesehatan yang tugas pokonya adalah menyelenggarakan pelayanan Kesehatan perorangan secara paripurna yang menyediakan pelayanan rawat inap, rawat jalan dan gawat darurat” Rahasia kedokteran atau rahasia medis merupakan hak pasien. Menjaga rahasia medis termasuk kewajiban bagi profesi rekam medis dan administrasi rumah sakit dalam menjalankan tugasnya di rumah sakit (Kurniawan , A. L., & Setiawan, 2021). Apabila rahasia rekam medis dan administrasi rumah sakit dibocorkan dapat dimintai pertanggung jawaban bagi pelanggarannya, khususnya pasal 332 kitab Undang-undang hukum pidana (KUHP). Pemerintah juga mengadakan dan mengatur sistem informasi masyarakat, seperti informasi kesehatan. Sistem untuk mengembangkan sistem informasi adalah pelaporan, pendataan, pembagian kasus, serta Kesehatan dan kejadian penyakit lainnya. Dengan adanya informasi kesehatan tersebut pemerintah meringankan masyarakat agar memperoleh hak akses terhadap pelayanan masyarakat. Hak dari informasi kesehatan merupakan hak dasar sosial yang bersumber dari hak asasi manusia. Sementara dalam pelayanan Kesehatan dikenal adanya hak bagi rahasia medis (medical secrecy). Kewajiban menyimpan rahasia pasien adalah salah satu tugas dokter, rekam medis, dan petugas administrasi rumah sakit untuk menjaga martabat dan hak pasien. Hak atas informasi medis, tindakan medis memiliki hak atas isi catatan medis begitu juga Kerugian pasien adalah bila tidak dapat pelayanan yang sangar memuaskan dan begitu pasien sangat meresahkan kerugian atas pelayanan yang sangat kurang memuaskan, seharusnya pihak rumah sakit menyediakan penanganan terhadap pasien agar merasa puas agar pasien tidak terasa kalau lagi sakit, kerugian pasien yang mengalami kebocoran data mengakibatkan pasien menjadi rugi contohnya sebagai pengajuan klaim asuransi palsu dan pencurian identitas. 2. TINJAUAN PUSTAKA Dalam penelusuran pustaka, peneliti menemukan banyak sekali teori konsep yang ada dalam berbagai literatur ilmiah yang sesuai dengan hal yang peneliti bahas yaitu mengenai Cycber crime dalam bidang kesehatan. Namun, dari banyaknya sumber literatur yang ada, ternyata kurang sekali penelitian yang menganalisis pengaruh Cyber crime terhadap data pasien. Oleh karena itu, penulis mengambil referensi beberapa konsep dari literatur ilmiah tersebut untuk dikembangkan dalam penelitian ini. 1. PENDAHULUAN Hak adalah wewenang (badan hukum) yang dimiliki seseorang untuk mendapatkan hak yang didapatkan pasien, Hak pasien yaitu menerima penjelasan tentang aturan rumah sakit, pelayanan medis, atau penyakit yang di derita pasien. Pasien berhak menerima pelayanan spesial dari pihak rumah sakit dan pasien berhak di rawat oleh dokter dan tanpa campur tangan pihak luar, dan pasien berhak mengajukan pengaduan atau komplain dari pihak rumah sakit dan pihak rumah sakit juga harus tetap menerima dengan lapang dada, bila menerima masukan atau komplain dari pasien dan pasien juga harus sabar dalam pelayanan karna pihak rumah sakit menangani pasien dengan penuh ikhlas dan sabar. Begitu juga pihak rumah sakit juga harus memberitahu kepada pasien tentang masalah kesehatan pasien atau yang sedang di derita pasien dengan halus, jelas, dan pasti, tanpa ada setingan dari rekam medis, begitu juga kita selaku pelaksanaan rumah sakit berusaha semaksimal mungkin untuk menjaga data diri pasien atas penderitaan yang pernah dialami di rumah sakit, karna di era sekarang sangat berbahaya berita dari luar tentang adanya kebobolan data diri pasien di media sosial atau hal ini sangat mempengaruhi identitas pasien dan pihak rumah sakit atas penyebaran nama baik. Karna di era sekarang banyak orang yang ingin memalsukan data pasien untuk hal yang tidak masuk akal. Rumah Sakit yakni suatu jenis sarana pelayanan kesehatan, memiliki tugas utama melayani Kesehatan perorangan serta tugas pelayanan lainnya. Meski dalam Undang-undang Kesehatan tidak menyebutkan secara jelas, namun pengertian Rumah Sakit bisa disimpulkan sebagai suatu bentuk fasilitas pelayanan kesehatan. Seperti dirumuskan pada Pasal 1 butir 7 bahwa “Fasilitas pelayanan kesehatan adalah suatu alat dan/atau tempat yang digunakan untuk menyelenggarakan upaya pelayanan kesehatan, baik promtif, preventif, kuratif maupun rehabilitatif yang dilakukan oleh pemerintah daerah dan masyarakat. Pengertian Rumah Sakit dirumuskan pada Pasal 1 butir 1 Undang-undang Nomor 44 Tahun 2009 tentang Rumah Sakit bahwa: “Rumah Sakit adalah institusi pelayanan Kesehatan perorangan secara paripurna yang menyediakan pelayanan rawat inap, rawat jalan, dan gawat darurat.” Vol. 01, No. 02, Terbit: pp. 47-53 Sanskara Hukum dan HAM (SHH) Sanskara Hukum dan HAM (SHH) 2.2 Pengaruh Cyber crime Cyber crime yang merupakan suatu kejahatan di Internet termasuk ke dalam tindakan kriminal. Hal ini menjadi ancaman serius bagi peradaban modern. Orang-orang menggunakan Internet untuk tujuan yang baik, tetapi peretas dan penjahat dunia maya menggunakan saluran yang sama untuk tujuan yang buruk. Ini telah menjadi masalah besar di zaman modern karena mempengaruhi semua orang di abad ke-21. telah menjadi lebih umum daripada sebelumnya. Kejahatan dapat dilakukan di mana saja orang terhubung melalui Internet. Orang dapat dengan mudah berkomunikasi satu sama lain tanpa menghirup udara. Ini memudahkan penjahat untuk melakukan kejahatan dan melarikan diri tanpa tertangkap. Selain itu, kejahatan dunia maya jauh lebih sulit dilacak daripada kejahatan fisik. Hal ini membuat sulit untuk mengadili penjahat dunia maya dan memulihkan properti yang dicuri. Pengaruh Cycber crime sendiri telah banyak dirasakan oleh masyarakat dalam berbagai sektor. Dari penelitian sebelumnya, telah dianalisis bagaimana pengaruh Cycber crime dalam sektor keuangan. Dalam sektor keuangan, Cycber crime sendiri telah meretas Cyber security compilance yang dilakukan oleh hacking (Kwarto F, Angsito M/UBM, 2018). Sedangkan dalam penelitian sekarang, analisis yang dilakukan adalah mengenai perlindungan data pribadi pasien terhadap serangan cycber crime. Kebaruan dari penelitian sebelumnya adalah analisis dari sektor yang berbeda yaitu sektor kesehatan sebagai upaya menanggulangi pengaruh Cycber crime dalam masyarakat. 2.1 Cycber crime Kejahatan dunia maya sangat meresahkan korban yang ditimpanya. Apalagi, sekarang banyak sekali jenis kejahatan itu sendiri. Salah satunya adalah Cycber crime yang dalam kasusnya telah meringkus data-data jutaan orang. Menurut Gregory (2005), Cyber crime adalah suatu bentuk kejahatan dunia maya dengan menggunakan media komputer yang terhubung ke internet, dan mengeksploitasi komputer lain yang terhubung dengan internet juga. Adanya lubang-lubang keamanan pada sistem operasi menyebabkan kelemahan dan terbukanya lubang yang dapat digunakan para hacker, cracker dan script kiddies untuk menyusup ke dalam komputer tersebut. Pendapat lain menyebutkan bahwa Cycber crime adalah kejahatan di mana tindakan kriminal hanya bisa dilakukan dengan Vol. 01, No. 02, Terbit: pp. 47-53 Sanskara Hukum dan HAM (SHH) r 50 teknologi cyber dan terjadi di dunia cyber (Fajri, dalam Tavani 2008). Dari kedua pendapat para ahli di atas dapat disimpulkan bahwa Cyber Crime adalah suatu kejahatan yang dilakukan menggunakan komputer kepada komputer lainnya yang saling terhubung ke internet demi mendapatkan keuntungan dari pihak tersebut. Walaupun tujuan utama adalah mendapatkan keuntungan berupa data, kadang alasan adanya Cycber crime ini juga tidak berdasar misalnya hanya ingin menyalahgunakan kecanggihan internet tanpa mengambil keuntungan. 4. HASIL DAN PEMBAHASAN Data pribadi merupakan segala informasi yang berkaitan dengan pribadi seseorang. Seiring dengan berjalannya waktu, perkembangan teknologi yang juga semakin canggih. Hal ini dapat memicu potensi terjadinya pelanggaran terhadap data pribadi yang menjadi semakin besar. Kegiatan kriminal yang dilakukan dengan menggunakan komputer atau jaringan komputer bertujuan sebagai alat atau sasaran pembobolan data disebut sebagai kejahatan dunia maya atau cyber crime (Aswandi, R., Muchsin, P. R., & Sultan, 2020). Pada prinsipnya hukum adalah pengaturan sikap (perilaku) seseorang dan masyarakat, menghukum bagi mereka yang melanggarnya. Meskipun dalam dunia maya, hukum juga harus ditegakkan untuk mengatur perilaku masyarakat. Pengaruh dari tindak kejahatan ini sangat berpengaruh terhadap kerugian masyarakat (Rumlus, M. H., & Hartadi, 2020). Undang-undang Nomor 11 Tahun 2008 tentang Informasi dan Transaksi Elektronik (ITE) yang menjadi landasan hukum untuk kasus kejahatan dunia maya. Dengan UU ITE, masyarakat pengguna informasi di Indonesia dapat terlindungi. Hal ini penting karena jumlah pengguna internet dan teknologi terus meningkat dari tahun ke tahun. Kemajuan teknologi sangat mempengaruhi pola pikir serta gaya hidup manusia, sebab itu banyak kasus kejahatan dengan menggunakan komputer atau jaringan teknologi (Oktaviani, S., Dewata, Y. J., & Fadlian, 2021). Di satu sisi dengan penggunaan internet dapat memberikan kemudahan bagi masyarakat dalam membantu aktivitas dan pekerjaannya. Disisi lain juga memudahkan pihak tertentu untuk melakukan suatu kejahatan kriminal (crybercrime). Karena cybercrime merupakan fenomena yang berkembang pesat dan tidak mengenal batas geografis, maka penting untuk diwaspadai karena berbeda dengan jenis kejahatan pada umumnya (Ersya, 2017). Seperti yang terjadi, kasus tahun lalu jutaan data dan informasi kesehatan penduduk Indonesia bocor, sekitar 279 juta data yang direkap oleh Badan Pengelola Jaminan Sosial Kesehatan (BPJS) juga bocor. Dari kasus tersebut menandakan bahwa keamanan data di Indonesia masih sangat rentan. Bagi pasien itu sendiri jika data pribadinya bocor seperti NIK, nomor hp, dan alamat rumahnya, ini pasti akan digunakan si pelaku untuk kejahatan seperti pinjaman online atau untuk klaim hak asuransi palsu, dan kalau pasien memiliki riwayat penyakit seperti HIV dan tersebar ke publik maka akan berisiko terkucilkan masyarakat karena penyakitnya tersebut. Bila masyarakat sudah mengetahui bahwa rumah sakit tersebut mengalami kebocoran data, sangat berpengaruh pada sektor pelayanan rumah sakit dan akan menimbulkan dampak besar pada rumah sakit tersebut. 2.3 Cycber Crime dalam Sektor Kesehatan Efek kejahatan dunia maya pada sektor kesehatan sangat mengkhawatirkan. Sektor kesehatan terkena dampak langsung dari kejahatan dunia maya dan juga merupakan salah satu sektor yang paling rentan terhadap kejahatan dunia maya. Dampak kejahatan dunia maya sangat serius dan dapat berdampak signifikan pada bidang kesehatan. Oleh karena itu, penting untuk memahami berbagai jenis kejahatan dunia maya dan bagaimana hal itu dapat berdampak negatif pada sektor kesehatan. Hal ini disebabkan penggunaan teknologi yang ekstensif, catatan pasien, dan keterlibatannya dengan bidang medis. Banyak institusi kesehatan menggunakan komputer untuk mengelola institusi mereka. Aplikasi utama yang digunakan oleh institusi kesehatan adalah spread sheet, email, pengolah kata, dan database. Semua aplikasi ini rentan terhadap kejahatan dunia maya dan dapat dibajak dengan mudah. Hal ini mempersulit institusi kesehatan untuk melindungi diri dari serangan online. Analisis yang dilakukan penelitian kali ini adalah mengenai perlindungan data pribadi pasien terhadap serangan cybercrime karena kejahatan dunia maya satu ini sangat perlu untuk ditanggulangi. Pada penelitian terdahulu, terdapat penelitian yang bertujuan untuk menganalisa upaya yang dilakukan pemerintah dalam rangka melindungi data pribadi khususnya terkait dengan rekam medis pasien Covid-19 terhadap bahaya kejahatan dunia maya seperti Cycber crime (Kurniawan, A.L, Setiawan A/UNS, 2021). Kebaruan dari penelitian ini dengan penelitian sekarang adalah perlindungan yang sebelumnya hanya diperuntukkan untuk pasien Covid-19 akan diimplementasikan untuk pasien konvensional sebagai langkah seribu menghadapi gencarnya dampak Cycber crime. Vol. 01, No. 02, Terbit: pp. 47-53 Sanskara Hukum dan HAM (SHH) r 51 3. METODE PENELITIAN Penelitian ini menggunakan metode penelitian hukum normatif yakni suatu proses guna menemukan suatu hukum terbuka, prinsip hukum, atau doktrin hukum untuk menjawab isu hukum yang dihadapi. Selain itu, juga digunakan untuk menghasilkan suatu argumentasi teori serta konsep yang baru dengan menganalisis beragam peraturan kaidah (norma), dan perundang- undangan sebagai objek. 5. KESIMPULAN Berdasarkan kesimpulan di atas data pasien pribadi mempunyai masalah pedoman yang terjadi kebocoran data pribadi pasien. Bagaimanapun juga data pasien yang bocor itu membuat masalah yang serius, bahayanya kebocoran data pasien bisa membahayakan keberadaan pasien. Berdasarkan analisis yang disampaikan dalam pembahasan maka kesimpulan yang bisa diambil yakni bahwa perlindungan data pasien atau data pribadi merupakan suatu hal yang sangat privasi jika data ini bocor akan berdampak dan risiko apabila terjadi kebocoran data pribadi pasien. Dampak kebocoran data rumah sakit, bagi rumah sakit data pasien akan menjadi kerugian ekonomi rumah sakit dan akan membuat nama dan kepercayaan kepada masyarakat menjadi rusak. Oleh karena itu, apabila terjadi penyalahgunaan data pribadi, untuk masyarakat sekitar, dan selalu disampaikan secara langsung dan diajukan kepada pihak yang berwenang. Sanskara Hukum dan HAM (SHH) r 52 r 5 Terdapat tantangan terhadap kebocoran data pasien terhadap celah kebocoran data pasien. Tidak diragukan, lebih mudah untuk mengakses data pribadi pasien ketika disimpan di ruang digital. Namun, karena ruang digital terbuka, kita harus memastikan keamanan digitalnya untuk mencegah diakses orang lain. Pentingnya melindungi data pribadi menjadikan suatu tantangan yang harus diatasi karena kurangnya kesadaran tersebut, terutama karena data kesehatan mencakup informasi yang sensitif (Rizkinaswara, 2020). Literasi harus digunakan untuk mendidik semua komponen, pemilik, pengontrol, dan prosesor. Pasien adalah pemilik data pribadi, Rumah sakit atau departemen kesehatan adalah pengontrol data pribadi dan penyedia layanan cloud adalah pemroses data. Menurut Dirjen Samuel menyatakan bahwa UU PDP telah ditetapkan telah ditetapkan, dengan ditetapkannya UU maka setiap institusi harus memiliki Data Protection Officer (DPO). Pesatnya perkembangan wearable devices dan health apps. Dirjen Samuel menjanjikan perangkat yang dapat dipakai dan aplikasi kesehatan yang juga memiliki potensi kebocoran data pribadi, dengan menerapkan pengaturan dan prosedur yang sama di rumah sakit dalam perlindungan data pribadi. Data kesehatan pasien merupakan data yang memiliki sifat rahasia sehingga tidak dapat dipublikasikan kecuali berdasarkan ketentuan yang telah diatur dalam Peraturan Perundang- Undangan. Penelitian ini bertujuan untuk mengetahui dampak serta risiko dari kebocoran data pribadi pasien. Dampak kebocoran data rumah sakit, bagi rumah sakit data pasien akan menjadi kerugian ekonomi rumah sakit dan akan membuat nama dan kepercayaan kepada masyarakat menjadi rusak. Oleh karena itu, apabila terjadi penyalahgunaan data pribadi, untuk masyarakat sekitar, dan selalu disampaikan secara langsung dan diajukan kepada pihak yang berwenang. Pengelolaan sendiri atau upaya pencegahan, jika peraturan yang berlaku tidak mencakup sistem penyalahgunaan data pribadi. Sejauh ini masyarakat mempunyai kebijakan dan ketentuan, selama yang diatur tersendiri dalam beberapa masalah yang mengatur tentang perundang- undangan, sehingga lebih lengkap, jelas, dan pasti. Saat ini, perlindungan data pribadi dilindungi oleh sejumlah undang-undang dan peraturan. Undang-undang ITE mengatur privasi, Pengaturan yaitu hanya orang berwenang dengan jalur hukum. Kelemahan itu muncul dari uraian pasal 26 UU ITE. Kurangnya dalam pasal tersebut yakni masalah kontraktor dan layanannya. 4. HASIL DAN PEMBAHASAN Jika tidak ada kebijakan masalah hal pembocoran ini tidak segera di lanjut tangan maka akan menjadikan ke tidak nyamanan pasien yang sedang di rawat, karena mereka takut jika data mereka akan tersebar bila tidak ada penanggung jawab terhadap pasien, menjadikan mereka tidak nyaman saat berkas mereka di minta pada pihak rumah sakit, untuk penanganan kasus ini harus di selesaikan dengan kesepakatan bersama dan tanggung jawab pihak rumah sakit serta menjadikan rumah sakit nyaman dan data pasien terpercaya (aman). Vol. 01, No. 02, Terbit: pp. 47-53 Sanskara Hukum dan HAM (SHH) Vol. 01, No. 02, Terbit: pp. 47-53 Aswandi, R., Muchsin, P. R., & Sultan, M. (2020). Perlindungan Data Dan Infoormasi Pribadi Melalui Vol. 01, No. 02, Terbit: pp. 47-53 DAFTAR PUSTAKA Vol. 01, No. 02, Terbit: pp. 47-53 r 53 Sanskara Hukum dan HAM (SHH) r Indonesian Data Protection System (IDPS). Legislatif, 167–190. Indonesian Data Protection System (IDPS). Legislatif, 167–190. Ersya, M. P. (2017). Permasalahan Hukum Dalam Menanggulangi Cyber Crime Di Indonesia. Journal Of Moral Use the “Insert Citation” Button to Add Citations to This Document. And Civic Education, 1(1), 50–62. Kurniawan , A. L., & Setiawan, A. (2021). Perlindungan Data Rekam Medis Sebagai Bentuk Perlindungan Data Pribadi Pasien Selama Pandemi Covid 19. Jurnal Hukum Dan Pembangunan Ekonomi, 9(1), 95–112. Oktaviani, S., Dewata, Y. J., & Fadlian, A. (2021). ertanggung Jawaban Pidana Kebocoran Data BPJS Dalam Perspeltif UU ITE. De Juncto Delicti: Journal Of Law, 1(2), 146–157. Rizkinaswara, L. (2020). Tantangan Perlindungan Data Pribadi Di Sektor Kesehatan. , ( ) g g Rumlus, M. H., & Hartadi, H. (2020). Kebijakan Penanggulangan Pencurian Data Pribadi Dalam Media Elektronik. Jurnal HAM, 285–299. J Santoso, A. A., Musta’in, & Utomo, E. K. (2021). Hukum Keperawatan. Trans Info Media. Santoso, A. P., Listyorini, P. I., Suyatno, A., Pujiyanto, R., & Oktaviani, I. (2021). Adminnistrasi Kebijakan Rumah Sakit. 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https://openalex.org/W4295441812	OpenAlex	Open Science	CC-By	2,022	Contactless Heartbeat Measurement Using Speckle Vibrometry	Shuhao Que	English	Spoken	6,799	11,946	Document status and date: Published: 08/09/2022 Document status and date: Published: 08/09/2022 Document Version: Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) Document Version: Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) DOI: 10.1109/EMBC48229.2022.9871712 DOI: 10.1109/EMBC48229.2022.9871712 Please check the document version of this publication: • A submitted manuscript is the version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. 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If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, please follow below link for the End User Agreement: Contactless Heartbeat Measurement Using Speckle Vibrometry Citation for published version (APA): Que, S., Verkruysse, W., van Gastel, M., & Stuijk, S. (2022). Contactless Heartbeat Measurement Using Speckle Vibrometry. In 2022 44th Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC) (pp. 4604-4610). Article 9871712 Institute of Electrical and Electronics Engineers. https://doi.org/10.1109/EMBC48229.2022.9871712 Citation for published version (APA): Que, S., Verkruysse, W., van Gastel, M., & Stuijk, S. (2022). Contactless Heartbeat Measurement Using Speckle Vibrometry. In 2022 44th Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC) (pp. 4604-4610). Article 9871712 Institute of Electrical and Electronics Engineers. https://doi.org/10.1109/EMBC48229.2022.9871712 Citation for published version (APA): Que, S., Verkruysse, W., van Gastel, M., & Stuijk, S. (2022). Contactless Heartbeat Measurement Using Speckle Vibrometry. In 2022 44th Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC) (pp. 4604-4610). Article 9871712 Institute of Electrical and Electronics Engineers. https://doi.org/10.1109/EMBC48229.2022.9871712 Download date: 25. Oct. 2024 Contactless Heartbeat Measurement Using Speckle Vibrometry Shuhao Que1, Willem Verkruijsse2, Mark van Gastel2 and Sander Stuijk1 Shuhao Que1, Willem Verkruijsse2, Mark van Gastel2 and Sander Stuijk1 Inside the speckle pattern, the granular spots of intensity that can be observed by using a defocused camera are called speckles. The speckle effect results from the interference among waves that share the same frequency but differ in phase and amplitude [10]. The laser speckle effect has long been investigated (e.g., estimation of ego-motion [11]) and SV has been used for various applications including speech [9] and heartbeat extraction [9] [12] [13]. SV en- ables magnification of micro-vibrations on the surface and therefore allows for heartbeat detection similar to Seismocar- diogram (SCG) [14] and Gyrocardiogram (GCG) [15]. Other popular motion based remote sensing technologies include laser Doppler vibrometry (LDV) and radio frequency (RF). LDV can detect vibration velocity of the surface where the laser spot is focused, by comparing the frequency shift be- tween the emitted and reflected laser beams [16]. Marchionni et al. [17] demonstrated the feasibility of non-contact mon- itoring of heart rate and respiration rate on preterm infants using LDV. Wang et al. [18] even managed to use pulsed laser vibrometer to monitor cardiac activity through clothes. RF can extract heartbeat by processing the phase variation information of the microwave radar signal [19]. Feasibility of heart rate extraction using RF has been demonstrated by [20] and [21]. Despite the contactless nature of LDV and RF for heartbeat monitoring, there are also downsides to consider. LDV can only detect the movement whose direction is along the incident laser beam [22] and the measured surface has to be reasonably reflective [23] [19] which often requires the use of retro-reflective materials on the skin [24] [25]. As for RF, the heartbeat-induced motion is very small compared to other detected motion, which renders isolation of the cardiac component difficult [26]. Furthermore, the positioning of the radar device and measuring distance is limited by limitations on power emission because of safety [26]. By comparison, SV does not require strict camera-surface positioning like LDV and no retro-reflective materials are needed since laser speckle patterns can be formed on any kind of optically rough surface (e.g., human skin and commonly used textiles). Besides, the use of a laser beam enables SV to measure at a further distance and be more precise and flexible in terms of region of interest selection. *This study was approved by the ethical review board of the Eindhoven University of Technology and funded by the NWO UMOSA project (17117). 1S. Que and S. Stuijk are with Eindhoven University of Technology, Eind- hoven, The Netherlands. s.que@tue.nl; s.stuijk@tue.nl. 2M. van Gastel and W. Verkruijsse are with Philips Research, Eind- hoven, The Netherlands. mark.van.gastel@philips.com; wim.verkruijsse@philips.com. This work is licensed under a Creative Commons Attribution 3.0 License. For more information, see http://creativecommons.org/licenses/by/3.0/ Contactless Heartbeat Measurement Using Speckle Vibrometry Shuhao Que1, Willem Verkruijsse2, Mark van Gastel2 and Sander Stuijk1 Remote Photoplethysmography (PPG) [27] is another con- tactless sensing technology for heart rate monitoring which Abstract— Monitoring of heart rate in patients in the general ward is necessary to assess the clinical situation of the patient. Currently, this is done via spot-checks on pulse rate manually or on heart rate using Electrocardiogram (ECG) by nurses. More frequent measurements would allow early detection of adverse cardiac events. In this work, we investigate a contactless measurement setup combined with a signal processing pipeline, which is based on speckle vibrometry (SV), to perform con- tactless heart rate monitoring of human subjects in a supine position, mimicking a resting scenario in the general ward. Our results demonstrate the feasibility of extracting heart rate with SV through varying textile thicknesses (i.e., 8 mm, 32 mm and 64 mm), with an error smaller than 3 beats per minute on average compared to the ground-truth heart rate derived from ECG. www.tue.nl/taverne Take down policy If you believe that this document breaches copyright please contact us at: openaccess@tue.nl providing details and we will investigate your claim. Download date: 25. Oct. 2024 I. INTRODUCTION In hospitalized patients, heart rate (or pulse rate) as one of the vital signs is measured to assess the clinical situation and possible deterioration of the patient [1] [2]. Currently, monitoring of heart rate in the general ward is done by nurses spot-checking pulse rate through pulse oximeter or heart rate through ECG every 4-6 hours [3]. Besides, according to a recent study [4], one-third of vital sign spot-checks are not done on time and one quarter are incomplete [2]. Such limited frequency might not be sufficient for early detection of adverse cardiac events since patients may experience deterioration between monitoring intervals [5] [2]. Thus, more frequent measurements are preferred. Current existing wearable devices have been proposed for continuous heart rate monitoring in the general ward, such as ViSi Mobile and HealthPatch [6]. Apart from ECG and pulse oximeter [2], other contact sensors also offer opportunity for general ward continuous heart rate monitoring, such as fabric-integrated multimode optical fiber [7] and finger plethysmography [8]. However, despite their wearable nature, these contact-based modalities still introduce inconvenience to patients’ move- ment. Therefore, in this work, we investigate the feasibility of using speckle vibrometry (SV) as a contactless alternative for more frequent heart rate measurement. SV technology can detect the motion-induced temporal variations of the laser speckle patterns [9]. A laser speckle pattern is essentially a random intensity distribution, pro- duced when a coherent laser beam is reflected from a surface that is optically rough on the scale of the laser wavelength. Remote Photoplethysmography (PPG) [27] is another con- tactless sensing technology for heart rate monitoring, which is based on a camera to detect blood volume changes on the skin. In the general ward, textiles (e.g., bedsheet) are commonly used by patients. Remote PPG requires visible skin and textiles might pose a risk that it occludes the skin. Therefore, using ECG as reference, this work aimed at validating the feasibility of using SV for contactless heart 4604 rate measurement through textiles while the subjects were lying on their back. We first conducted on-skin measurements on four selected anatomical locations (i.e., sternum, tricuspid, aortic and central abdomen) and selected tricuspid, which achieved the best performance with on average 0.3 beats of difference per minute compared with ECG, as the best aiming location for through-textile measurements. A. SV and Motion Extraction Our setup is based on a front-defocused camera system to register the motion of the imaged speckle pattern. Following the theoretical explanations presented by Zalevsky et al. in [9], when the camera is defocused, the imaging plane is shifted from the detected object to a plane positioned at a distance L1 (see Figure 1) and L2 is the distance between the camera focus plane and the camera lens. Under this imaging condition, neither transversal nor axial movement affects the distribution of the speckle pattern [9]. The tilt of the detected object manifests as displacements of the speckle pattern imaged in the camera, as presented by the following equation: Y X Laser Camera & Lens L1 L2 Fig. 2. Experimental setup for SV heart rate measurement. The blue rectangle indicates the positioning of the camera and the laser (in the green rectangle) is attached to the camera lens in an aligned fashion. The red arrows x and y indicate the x- and y-axes in the camera field of view, which also correspond to the two directions of the SV measurement. d = L1F L2 tan α (1) (1) where d denotes a relative shift of the speckle pattern due to object tilt, α denotes the tilt angle of the object and F denotes the focal length of the imaging lens. The shift of the speckle pattern is proportional to the amount of camera defocus (i.e. L1 L2 ) [28]. 2 Video recordings of the laser speckle pattern were pro- cessed by a motion extraction algorithm to obtain heartbeat signals. In this work, we used the sub-pixel image regis- tration algorithm proposed by Guizar-Sicairos et al. [29], where sub-pixel translations of the speckles along both x- and y-axes were obtained by first locating the cross-correlation peak using a coarse-to-fine up-sampling approach and then dividing the coordinates of the peak by the up-sampling factor. The values of L1, L2 and F in our experimental setup were about 0.6 m, 0.4 m, and 0.05 m respectively, under which condition, we selected the up-sampling factor as 100, which indicates a sub-pixel accuracy of 0.01 pixels Fig. 2. Experimental setup for SV heart rate measurement. The blue rectangle indicates the positioning of the camera and the laser (in the green rectangle) is attached to the camera lens in an aligned fashion. I. INTRODUCTION The ob- tained results showcase the feasibility of using SV to monitor heart rate at several spots on the skin including one that is relatively distant from the heart (i.e., central abdomen) and one spot (i.e., tricuspid area) through textiles. that guarantees the detection of the minimum angular change represented as 0.00013 pixels/mm. The up-sampling factor of 100 was empirically selected to be sufficient to detect the motions induced by heartbeats. II. METHODS II. METHODS B. Experimental Setup and Protocols The experimental setup of SV for contactless heartbeat monitoring is shown in Figure 2. A monochrome camera with a 2.35 megapixel CMOS sensor (UI-3060CP-M-GL, IDS Imaging Development Systems GmbH) was used. An Aixiz Class-I green laser (AD-532-1-830) was mounted to a C-mount camera lens (M111FM50, Tamron) in parallel. The focus distance L2 of the camera lens was set to the minimal value of 0.4 m (see Figure 2). The laser and camera view were along the z-axis. The distance between the camera and the bed surface was 125 cm, rendering the measuring distance approxmately 100 cm. For SCG/GCG signal acquisition, an inertial measurement unit (BWT901CL, WitMotion) was firmly attached to the subjects’ sternum using double-sided skin tape (Nitto, ST-502-1). For the single-lead ECG signal acquisition, a Mind Media Nexus-10 was used with the BioTrace+ NX10 software package. This configuration was used in all experiments. Data from different devices was synchronized by introducing motion artifacts at the beginning of the measurements, yielding an estimated misalignment range from 0 ms to 150 ms. A. SV and Motion Extraction CoƩon Bedsheet (1 mm / layer) Furry Blanket (4 mm / layer) CoƩon Comforter (32 mm / layer) Fig. 4. Textile materials used for the study. (3) where M denotes the median and N denotes the number of x- and y-coordinate pairs of the whole recording in the selected Cartesian quadrant C. The same procedure was applied to x- and y-axes of GCG to obtain Gr for a fair waveform comparison with Dr. However, x- and y-axes of SCG appeared to be too noisy so only the z-axis Az was used for waveform comparison. 2) Peak detection: To detect the peaks in the SCG, GCG and SV signals, we propose the following algorithm consisting of three steps. Steps 1 and 2 were adopted from Tadi et al. [30]: Fig. 4. Textile materials used for the study. are presented in Table I. The detailed recording procedure for each subject is indicated in Table II. It is worth mentioning that the textiles were folded tightly and symmetrically once, once and three times to respectively obtain 2 layers of comforters, 2 layers of blankets and 3 layers of bedsheets. 8 and 16 layers of blankets were thus obtained by stacking 4 and 8 pieces of folded blankets respectively. 12 and 24 layers of bedsheets were thus obtained by stacking 4 and 8 pieces of folded bedsheets respectively. While each subject remained static in bed, the textiles were directly placed on them covering their whole upper torso without additional adjustment. For each separate recording, the textiles were removed and placed back on the subject. • Step 1: the signal was first passed through a fourth-order Butterworth IIR high-pass filter with a cutoff frequency equal to 0.5 Hz, with the purpose of removing baseline wander and respiratory components. The Hilbert transform was applied to the high-pass filtered signal to obtain its analytic signal. Since Hilbert transform yields a 90◦phase shift, the magnitude of the analytic signal was computed to derive the Hilbert amplitude envelope. • Step 2: the computed Hilbert amplitude envelope was then passed through a third-order Butterworth IIR band-pass filter with cutoff frequencies of 0.5 Hz and 3 Hz to obtain a simpler structured waveform that contains the same periodicity as the original signal, named as the filtered Hilbert envelope signal. 1https://docs.scipy.org/doc/scipy/reference/ generated/scipy.signal.find_peaks.html A. SV and Motion Extraction Although the participants were all healthy with a normal resting heart rate range (60-100 beats per minute), the lower bound 0.5 Hz (i.e., 30 beats per minute) and the upper bound 3 Hz (i.e., 180 beats per minute) were chosen since in the general ward patients might exhibit abnormal heart rates. A. SV and Motion Extraction The red arrows x and y indicate the x- and y-axes in the camera field of view, which also correspond to the two directions of the SV measurement. The Ethical Review Board (ERB) of the Eindhoven Uni- versity of Technology approved the data collection and processing protocol (ERB2020EE3) for this study. After obtaining their informed consent forms, a total of 9 healthy human subjects participated in the study (five males). Data collection was conducted on each subject while the subject was breathing spontaneously in a supine position. An in- dication of the selected anatomical locations is shown in Figure 3. The used textile materials are shown in Figure 4. The specifications of the devices used for the experiments Lens Sensor Imaged laser speckle pattern L1 L2 laser beam camera focus plane α d F speckle motion signal L1 L2 Laser beam Camera focus plane time ..... Fig. 1. Schematic of defocused camera system for speckle vibrometry. Fig. 1. Schematic of defocused camera system for speckle vibrometry. 4605 aortic sternum (SV) sternum (SCG/GCG) tricuspid central abdomen Fig. 3. Indication of the selected anatomical locations. CoƩon Bedsheet (1 mm / layer) Furry Blanket (4 mm / layer) CoƩon Comforter (32 mm / layer) Fig. 4. Textile materials used for the study. aortic sternum (SV) sternum (SCG/GCG) tricuspid central abdomen Fig. 3. Indication of the selected anatomical locations. aortic sternum (SV) sternum (SCG/GCG) tricuspid central abdomen For each one-minute recording of SV measurements, to locate one polar angle value that represents the dominant vibration direction of the heartbeat induced peaks, the am- plitudes calculated by q D2x + D2y were derived in four Cartesian quadrants respectively. The Cartesian quadrant with the highest 90th percentile values of the amplitudes were selected and the dominant vibration angle was the median of all existing θ values within this plane, denoted by ˆθ. The derivation of ˆθ is presented below:              C = argmaxCj{P90( q D2x + D2y)[(x,y)∈Cj]}, Cj = j-th Cartesian quadrant θi = arctan Dyi[(xi,yi)∈C] Dxi[(xi,yi)∈C] i = 1,2,3,...,N ˆθ = M({θi}) (3) Fig. 3. Indication of the selected anatomical locations. CoƩon Bedsheet (1 mm / layer) Furry Blanket (4 mm / layer) CoƩon Comforter (32 mm / layer) Fig. 4. Textile materials used for the study. Dr = Dx · cosˆθ + Dy · sinˆθ (2) C. Evaluation Metrics L2 (m) 0.4 TABLE II RECORDING PROCEDURE ON EACH SUBJECT Anatomical location Textile material Layer thickness Number of layers Duration of Repetitions per Number of one recording subject subjects Sternum N/A 0 mm 0 1 minute 3 9 Tricuspid N/A 0 mm 0 1 minute 3 9 Aortic N/A 0 mm 0 1 minute 3 9 Central abdomen N/A 0 mm 0 1 minute 3 9 Tricuspid N/A 0 mm 0 1 minute 3 9 Tricuspid area Furry blanket 8 mm 2 1 minute 3 9 Tricuspid area Furry blanket 32 mm 8 1 minute 3 9 Tricuspid area Furry blanket 64 mm 16 1 minute 3 9 Tricuspid area Cotton sheet 8 mm 3 1 minute 3 9 Tricuspid area Cotton sheet 32 mm 12 1 minute 3 9 Tricuspid area Cotton sheet 64 mm 24 1 minute 3 9 Tricuspid area Cotton comforter 32 mm 1 1 minute 3 9 Tricuspid area Cotton comforter 64 mm 2 1 minute 3 9 TABLE I Anatomical location Textile material Layer thickness Number of layers Duration of Repetitions per Number of one recording subject subjects Sternum N/A 0 mm 0 1 minute 3 9 Tricuspid N/A 0 mm 0 1 minute 3 9 Aortic N/A 0 mm 0 1 minute 3 9 Central abdomen N/A 0 mm 0 1 minute 3 9 Tricuspid N/A 0 mm 0 1 minute 3 9 Tricuspid area Furry blanket 8 mm 2 1 minute 3 9 Tricuspid area Furry blanket 32 mm 8 1 minute 3 9 Tricuspid area Furry blanket 64 mm 16 1 minute 3 9 Tricuspid area Cotton sheet 8 mm 3 1 minute 3 9 Tricuspid area Cotton sheet 32 mm 12 1 minute 3 9 Tricuspid area Cotton sheet 64 mm 24 1 minute 3 9 Tricuspid area Cotton comforter 32 mm 1 1 minute 3 9 Tricuspid area Cotton comforter 64 mm 2 1 minute 3 9 signal to locate the original peak locations through window-searching of local maximal amplitudes. The range of the i-th window was defined as [0.5∗(peaki − peaki−1), peaki+A], where A was empirically selected to be 150 ms to avoid locating the peak of the next cardiac cycle. from two subjects which show clear raw waveforms. ECG demonstrates the most stable peak morphology among all. C. Evaluation Metrics Firstly, we proposed a method to derive a single measure- ment for SV. Secondly, using ECG as a reference, combined with two peak detection algorithms that are presented below, average heart rates (HRa) were derived from the beat-to-beat intervals of the ECG and SV signals for comparison and quality assessment. 1) Derivation of SV measurement Dr: A SV signal has two directions, i.e., along its x- and y-axes. These are denoted as Dx and Dy (see Figure 2) in this work. By mapping the Cartesian coordinates to polar coordinates, we located the dominant heartbeat-induced vibration direction, denoted by ˆθ, and thereby we derived a single measurement for SV denoted by Dr, which is independent of the camera-surface orientation. The conversion equation is presented below: • Step 3: we employed the find_peaks1 function from the Scipy library of Python to perform peak detection on the filtered Hilbert envelope signal with the required minimal peak height set as 0. The envelope peaks, i.e., the detected peak locations of the filtered Hilbert envelope signal, were then mapped back to the original Dr = Dx · cosˆθ + Dy · sinˆθ (2) Dr = Dx · cosˆθ + Dy · sinˆθ (2) 4606 4606 TABLE I DEVICE SPECIFICATIONS Camera & Lens Laser Accelerometer ECG Positioning of camera & laser Frame rate (fps) 360 Wavelength (nm) 532 Fs (Hz) 200 Fs (Hz) 256 Elevation (degrees) 90 Exposure (ms) 2.002 Power (mW) ≤1 Acc (g) ±2 Nr. of leads 1 Distance (cm) 100 Resolution (pixels) 280×330 Gyr (degree/s) ±250 Aperture (f-stop) F/1.8 Focus dist. (4) (4) 4) Average peak amplitude of Dr: By employing the peak detection algorithm mentioned above, both the maximal positive and negative amplitudes of SV measurements were determined within each search window formulated in Step 3. The average peak amplitude was thus derived from the average of three repeated measurements from each subject. B. Feasibility of SV Heart Rate Measurement Figure 8 illustrates the distributions of AHREa between ECG and on-skin SV signals calculated from all 9 subjects. It can be observed that for all anatomical locations, the median AHREa are less than 1 BPM with the largest deviation less than 3 BPM, indicating the feasibility of using SV to monitor heart rate on uncovered skin. SV signal on both tricuspid and aortic yielded the lowest AHREa while central abdomen the highest, but these differences were not significant. Figure 9 C. Evaluation Metrics In the upper plot (i.e., subject 5), the peak morphology of SV measurement Dr is most similar to that of GCG measurement Gr, both exhibiting low variability. In the lower plot (i.e., subject 2), the peak morphology of Dr is also most similar to that of Gr, both exhibiting larger variability. As for the R-peak detection of ECG signals, the imple- mentation of the widely adopted algorithm proposed by W. Engelse and C. Zeelenberg [31] with the modifications pro- posed by A. Lourenco et al. [32] was used, which proved to be capable of correctly detecting noisy single heartbeats [32]. For through-textile measurement, Figure 6 shows 10- second segments from subject 2 with respect to different textile materials and layer thicknesses. Subject 2 was se- lected because the collected measurements in all textile coverage scenarios exhibited clear raw waveforms. It can be observed that textile layers dampened the amplitudes of SV measurements. Figure 7 further shows that for most subjects, regardless of the textile materials, textile layers have dampening effect on the amplitudes of SV measurements in general. Exceptions are with 8 mm of bedsheet (subjects 4, 7, and 8). We speculate that 8 mm of bedsheet can enhance high-frequency heartbeat motions and dampen low-frequency respiratory ones in certain cases, which are probably de- pendent on body mass index (BMI), respiration-to-heartbeat magnitude ratio etc. and require further investigation. p y g y g 3) Average absolute heart rate errors: Using ECG as reference, the average absolute heart rate error (AHREa) of each subject’s three repeated measurements can be calculated from the following equation: AHREa = P3 i=1 \|HRa(ECGi) −HRa(SVi)\| 3 A. Waveform of SV Measurement No statistical significance was found according to Kruskal- Wallis H test (p-value=0.126). blanket bedsheet comforter Average Positive and Negative Amplitudes [pixels] Subject ID Pixels 0 mm 8 mm 32 mm 64 mm Fig. 7. Average amplitudes of the positive and negative peaks in the raw waveform of Dr from all 9 subjects. BPM Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 10. Average absolute heart rate error (BPM) of on-skin heartbeat measurement: comparison between males and females. The numbers of data points for each anatomical location are 16 and 20 for females and males respectively. No statistical significance was found according to Kruskal- W lli H t t ( l 0 126) blanket bedsheet comforter Average Positive and Negative Amplitudes [pixels] Subject ID Pixels 0 mm 8 mm 32 mm 64 mm Fig. 7. Average amplitudes of the positive and negative peaks in the raw waveform of Dr from all 9 subjects. blanket bedsheet comforter Average Positive and Negative Amplitudes [pixels] Subject ID Pixels 0 mm 8 mm 32 mm 64 mm Fig. 7. Average amplitudes of the positive and negative peaks in the raw waveform of Dr from all 9 subjects. Distribution of the Average Absolute Heart Rate (BPM) Error Average Positive and Negative Amplitudes [pixels] Average Positive and Negative Amplitudes [pixels] BPM Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 10. Average absolute heart rate error (BPM) of on-skin heartbeat measurement: comparison between males and females. The numbers of data points for each anatomical location are 16 and 20 for females and males respectively. No statistical significance was found according to Kruskal- Wallis H test (p-value=0.126). Fig. 7. Average amplitudes of the positive and negative peaks in the raw waveform of Dr from all 9 subjects. demonstrates that for the four anatomical locations, despite the difference in body shape between the two genders around the chest area, no significant differences were found in the distributions of AHREa. If we combine the four anatomi- cal locations, as shown in Figure 10, still no statistically signifcant difference was found between two genders (p- value=0.126). At each anatomical location, there is one male subject that exhibits higher AHREa values. We speculate that it was because of the subject’s BMI (>25), which might have negative impact on heart rate estimation based on SV. A. Waveform of SV Measurement Figure 11 illustrates that compared with on-skin SV, all three textile materials did not introduce significant differ- ences with 8 mm or 32 mm of layer thickness. However, when the layer thickness was 64 mm, bedsheet layers yieled slight increase in both median AHREa and interquartile range (IQR) values (p-value=0.031). Both comforter and blanket layers yielded significant increases (p-value=0.031, p-value=0.012). Therefore, it can be concluded that SV can provide accurate heart rate measurements even through 64 mm of bedsheets with median AHREa less than 2 BPM, which should suffice for through-textile measurement in the general ward. However, if the textile material was a blanket or a comforter, SV was less accurate when the layers have a thickness of 64 mm and exhibited larger values of both me- dian AHREa (i.e., more than 2 BPM) and IQR, where blanket layers yielded the worst performance. The blanket material used in the experiments was the softest, which potentially made it more predisposed to relative displacements between the skin surface and the textiles and that could hinder the propagation of vibration from the heart to the textile layer surface. tricuspid aortic sternum central abdomen BPM N = 9 Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 8. Average absolute heart rate error (BPM) of on-skin heartbeat measurement. The number of data points for each anatomical location is 9 (i.e., N = 9), composed by the average results of 3 repetitions from all 9 subjects. No statistical significance was found among these distributions according to Kruskal-Wallis H test [33]. tricuspid aortic sternum central abdomen BPM N = 9 Distribution of the Average Absolute Heart Rate (BPM) Error Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 8. Average absolute heart rate error (BPM) of on-skin heartbeat measurement. The number of data points for each anatomical location is 9 (i.e., N = 9), composed by the average results of 3 repetitions from all 9 subjects. No statistical significance was found among these distributions according to Kruskal-Wallis H test [33]. 0 mm 8 mm 32 mm 64 mm layer thickness N = 9 BPM Fig. 11. Average absolute heart rate errors (BPM) of through-textile heartbeat measurement. The number of data points for each textile cov- erage scenario is 9 (i.e., N = 9), composed by the average results of 3 repetitions from all 9 subjects. A. Waveform of SV Measurement A. Waveform of SV Measurement Since both GCG and SCG signals were obtained from the sternum, we chose SV measurements Dr collected from the sternum for comparison, along with ECG. Figure 5 displays the ensemble peaks from these four measurements collected 4607 Ensemble Peaks Normalized Time Axis [a.u.] Normalized Y-Axis [a.u.] SV: Dr GCG: Gr SCG: Az ECG Subject 5 Subject 2 ECG SV: Dr GCG: Gr SCG: Az Normalized Y-Axis [a.u.] Fig. 5. Ensemble peaks of ECG, on-sternum SV, GCG and SCG from subjects 5 and 2. The corresponding measurements were synchronised based on motion artifacts with peak misalignment less than 150 ms. Each graph is composed by 50 segments sampled from one continuous measurement. Ensemble Peaks Normalized Time Axis [a.u.] Normalized Y-Axis [a.u.] SV: Dr GCG: Gr SCG: Az ECG Subject 5 Subject 2 ECG SV: Dr GCG: Gr SCG: Az Normalized Y-Axis [a.u.] Normalized Y-Axis [a.u.] Fig. 5. Ensemble peaks of ECG, on-sternum SV, GCG and SCG from subjects 5 and 2. The corresponding measurements were synchronised based on motion artifacts with peak misalignment less than 150 ms. Each graph is composed by 50 segments sampled from one continuous measurement. 0 2 4 6 8 10 no layer blanket bedsheet blanket bedsheet blanket bedsheet comforter comforter 2 0 -2 2 0 -2 2 0 -2 0 2 4 6 8 10 0 2 4 6 8 10 1 0 -1 1 0 -1 2 2 0 -2 -4 1 0 -1 1 0 -1 0.50 0.25 0.00 -0.25 -0.50 Pixels Time [s] 10-Second Segments of Raw Through-Textile Dr of One Subject 0 2 4 6 8 10 0 mm 8 mm 32 mm 64 mm Fig. 6. 10-second segments of raw Dr of subject 2. The proposed peak detection algorithm was applied to all these segments. The y-axis of each individual plot denotes pixels and the x-axis denotes time in seconds. Fig. 6. 10-second segments of raw Dr of subject 2. The proposed peak detection algorithm was applied to all these segments. The y-axis of each individual plot denotes pixels and the x-axis denotes time in seconds. 4608 BPM Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 10. Average absolute heart rate error (BPM) of on-skin heartbeat measurement: comparison between males and females. The numbers of data points for each anatomical location are 16 and 20 for females and males respectively. A. Waveform of SV Measurement According to Kruskal-Wallis H test, statistical significance was found between 0 mm and 64 mm (blanket: p-value=0.012; bedsheet: p-value=0.031; comforter: p-value=0.031). 0 mm 8 mm 32 mm 64 mm layer thickness N = 9 BPM tricuspid aortic sternum central abdomen \|ECG - Dr\| BPM Distribution of the Average Absolute Heart Rate (BPM) Error Fig. 9. Average absolute heart rate error (BPM) of on-skin heartbeat measurement: comparison between males and females. The numbers of data points for each anatomical location are 4 and 5 for females and males respectively. No statistical significance was found according to Kruskal- Wallis H test. tricuspid aortic sternum central abdomen \|ECG - Dr\| BPM Distribution of the Average Absolute Heart Rate (BPM) Error Distribution of the Average Absolute Heart Rate (BPM) Error Distribution of the Average Absolute Heart Rate (BPM) Error 32 mm layer thickness Fig. 11. Average absolute heart rate errors (BPM) of through-textile heartbeat measurement. The number of data points for each textile cov- erage scenario is 9 (i.e., N = 9), composed by the average results of 3 repetitions from all 9 subjects. According to Kruskal-Wallis H test, statistical significance was found between 0 mm and 64 mm (blanket: p-value=0.012; bedsheet: p-value=0.031; comforter: p-value=0.031). Fig. 9. Average absolute heart rate error (BPM) of on-skin heartbeat measurement: comparison between males and females. The numbers of data points for each anatomical location are 4 and 5 for females and males respectively. 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CONCLUSION With proper on-skin selection of anatomical locations, SV is able to provide accurate heartbeat measurements through 4609 textiles within 3 BPM error on average, serving as a contact- less alternative to ECG in the general ward. Our current study is limited to the supine resting position and the anatomical location is limited to tricuspid area in case of through-textile measurement. Therefore, future work needs to investigate if SV still works when the resting position is lateral and if other anatomical locations that are relatively distant from the heart (e.g., central abdomen) can also provide through-textile heartbeat measurement. Subjects were asked to remain still during recording, where motion distortions that can happen in the general ward were not taken into consideration. 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https://openalex.org/W2162619348	OpenAlex	Open Science	CC-By	2,012	The scientific basis for a satellite mission to retrieve CCN concentrations and their impacts on convective clouds	Daniel Rosenfeld	English	Spoken	16,756	28,342	Correspondence to: D. Rosenfeld (daniel.rosenfeld@huji.ac.il) Received: 25 January 2012 – Published in Atmos. Meas. Tech. Discuss.: 10 February 2012 Revised: 21 July 2012 – Accepted: 25 July 2012 – Published: 23 August 2012 Abstract. The cloud-mediated aerosol radiative forcing is widely recognized as the main source of uncertainty in our knowledge of the anthropogenic forcing on climate. The current challenges for improving our understanding are (1) global measurements of cloud condensation nuclei (CCN) in the cloudy boundary layer from space, and (2) disentangling the effects of aerosols from the thermodynamic and meteo- rological effects on the clouds. Here, we present a new con- ceptual framework to help us overcome these two challenges, using relatively simple passive satellite measurements in the visible and infared (IR). The idea is to use the clouds them- selves as natural CCN chambers by retrieving simultaneously the number of activated aerosols at cloud base, Na, and the cloud base updraft speed. The Na is obtained by analyzing the distribution of cloud drop effective radius in convective elements as a function of distance above cloud base. The cloud base updraft velocities are estimated by double stereo- scopic viewing and tracking of the evolution of cloud surface features just above cloud base. In order to resolve the vertical dimension of the clouds, the ﬁeld of view will be 100 m for the microphysical retrievals, and 50 m for the stereoscopic measurements. The viewing geometry will be eastward and 30 degrees off nadir, with the Sun in the back at 30 degrees off zenith westward, requiring a Sun-synchronous orbit at 14 LST. Measuring simultaneously the thermodynamic environ- ment, the vertical motions of the clouds, their microstructure and the CCN concentration will allow separating the dynam- ics from the CCN effects. This concept is being applied in the proposed satellite mission named Clouds, Hazards and Aerosols Survey for Earth Researchers (CHASER). the proposed satellite mission named Clouds, Hazards and Aerosols Survey for Earth Researchers (CHASER). 1.1 Uncertainty in aerosol cloud-mediated forcing drives the climate uncertainty The effects of aerosols on clouds are recognized by the IPCC (2007) as the largest sources of uncertainty in the quantiﬁ- cation of anthropogenic perturbations to the Earth’s energy balance. Therefore, the reduction of this uncertainty is neces- sary for improving our understanding of the present climate and for the reduction of the uncertainties in the prediction of future climates (e.g., Andreae et al., 2005; Kiehl, 2007). The uncertainty in aerosol cloud-mediated radiative forcing is composed of two possibly large and highly uncertain op- posite effects from shallow and deep clouds (Rosenfeld et al., 2012a). This underlines the importance of conducting global measurements of the aerosol effects on clouds. This paper introduces a new concept for retrieving from space micro- physical and dynamical properties of convective clouds and reconstructs from that the cloud condensation nuclei (CCN), the vertical motions of the cloud tops, and the height at which rain and cloud glaciations initiate. This will allow disentan- gling the effects of aerosols (CCN and ice nuclei) and me- teorology (cloud vertical motion, temperatures and depth) on The scientiﬁc basis for a satellite mission to retrieve CCN concentrations and their impacts on convective clouds D. Rosenfeld1, E. Williams2, M. O. Andreae3, E. Freud1, U. P¨oschl3, and N. O. Renn´o4 1Institute of Earth Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel 2Parsons Laboratory, Massachusetts Institute of Technology (MIT), 77 Massachusetts Avenue, Cambridge, MA, 02139, USA 3Biogeochemistry Department, Max Planck Institute for Chemistry, P.O. Box 3060, 55020 Mainz, Germany 4Department of Atmospheric, Oceanic and Space Sciences, University of Michigan, 1531C Space Research Building, 2455 Hayward St., Ann Arbor, MI 48109, USA Correspondence to: D. Rosenfeld (daniel.rosenfeld@huji.ac.il) Correspondence to: D. Rosenfeld (daniel.rosenfeld@huji.ac.il) Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ doi:10.5194/amt-5-2039-2012 © Author(s) 2012. CC Attribution 3.0 License. Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ doi:10.5194/amt-5-2039-2012 © Author(s) 2012. CC Attribution 3.0 License. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations The aerosols were found even to affect the potential of deep convective clouds to produce large hail and tornadoes, where clouds with added pollution aerosols become potentially more damaging (Rosenfeld and Bell, 2011 and references therein). The impact of aerosols on cloud invigoration and the vertical distribution of latent heating can also modulate regional and large scale circula- tion systems, as observed by Bell et al. (2008) and simulated by Fan et al. (2012). p On a more technical level, this study focuses on the way by which combining a large number of various satellite- retrieved properties can provide new fundamentally impor- tant retrieved entities, while doing it on a sufﬁciently high resolution that can resolve the smallest convective elements. Characterization of the exact performance of retrieving each of the input-retrieved parameters constitutes a major study in its own right, which requires simulation and validation at the relevant scales. This pertains to properties such as cloud sur- face temperature, cloud particle effective radius, cloud phase, precipitable water above the cloud top, cloud surface verti- cal motion, and relation between cloud top rising rate and cloud updraft. This exact characterization has not yet been achieved. The realization of the kind of beneﬁts that can be realized when these parameters are combined in the frame- work presented here will hopefully serve as a motivation for the additional studies required for the exact characterization. For error calculations of these properties and for calculat- ing the error propagation through the high level products, we will use references where available or some estimates based on theoretical considerations. 1.2 Ways by which aerosols affect cloud microphysical, precipitation and radiation properties Aerosols are extremely important to cloud radiative forcing because cloud droplets are nucleated on pre-existing aerosols that serve as cloud condensation nuclei (CCN) and ice nuclei. The cloud droplet concentrations, Nd, that form near cloud base depend on the supersaturation (S) activation spectrum, CCN(S), and on the cloud base updraft speed, wb. The CCN that actually nucleate cloud droplets in speciﬁc clouds are re- ferred to as activated CCN, and their number density before dilution by mixing with ambient non-cloudy air is denoted as Na. The mixing with ambient air reduces Na to Nd, as long as no new droplets are nucleated. Such new nucleation can occur well above cloud base when cloud droplets are de- pleted by conversion into precipitation and the supersatura- tion increases (Pinsky and Khain, 2002). This is rarely an issue in non-precipitating boundary layer clouds. This num- ber determines the variability of the fundamental properties of the clouds in a given meteorological situation. The albedo of clouds with a given value of vertically-integrated liquid water path increases with Nd (Twomey, 1977). With greater Nd smaller droplets are formed, reducing coalescence and inhibiting rain production (Gunn and Phillips, 1957; Squires, D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2040 the properties of the observed cloud and precipitation proper- ties. A satellite mission for measuring the cloud properties is described in Renno et al. (2012). The full characterization of the cloud–aerosol–precipitation interactions can be achieved when combined with precipitation properties observed by other satellites or ground based measurements. Here, we pro- vide the scientiﬁc basis for these kinds of measurements from space, showing the feasibility of what has been considered impossible until now. 1958). The dynamic response to the rain suppression length- ens the life-time and increases the cloud cover when sup- pressing precipitation in clouds, at least in the case of shal- low heavily drizzling marine stratocumulus (Albrecht, 1989; Rosenfeld et al., 2006; Lebsock et al., 2008; Wang et al., 2011; Goren and Rosenfeld, 2012). In contrast, adding CCN to non-precipitating clouds can enhance their evaporation and mixing with the ambient air due to the decrease in cloud drop size (e.g., Wood, 2007; Jiang et al., 2009; Chen et al., 2011). In deep convective clouds rain forms eventually, but the aerosol-induced delay in its formation to greater heights was shown to cause in some conditions cloud invigoration (Andreae et al., 2004; Rosenfeld et al., 2008a; Fan et al., 2012) and additional electriﬁcation (Yuan et al., 2011). The invigoration was shown to occur mainly for situations with weak wind shear and for clouds with warm base, in which there is large vertical distance between cloud base and the freezing level (Fan et al., 2009; Li et al., 2011). The aerosol- induced vertical growth and the consequent expansion of the anvils into cirrus was observed (Koren et al., 2010) and simu- lated (Fan et al., 2012) to inﬂict large positive radiative forc- ing, in contrast to the strong negative forcing that is caused by the aerosol effect on shallow clouds, at least in heavily drizzling marine stratocumulus (e.g., Albrecht, 1989). In the context of tropical cyclones, the invigoration of clouds at the periphery of the storms occurs at the expense of a reduc- tion in the amount of air converging into the storm center, hence reducing the maximum wind speeds (Rosenfeld et al., 2012b and references therein). Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2041 from satellite observations where aerosol optical properties are compared to cloud radiative properties. This practice is inaccurate, mainly because of the poor ability to convert the aerosol optical depth (AOD) into concentrations of cloud condensation nuclei (CCN) that actually interact with the clouds (Andreae, 2009). The inaccuracies are caused by the fact that aerosols are not measured at the same height as the cloud base, cloud contamination (i.e., clouds enhancing the aerosol signals), and aerosol swelling in the moist envi- ronment near the clouds (Koren et al., 2007). Furthermore, aerosols in the cloudy boundary layers are often obscured by the clouds so that there is no way to measure them directly re- gardless of these issues with accuracy. This obviously occurs in conditions where the importance for measuring them is greatest because they interact with the clouds that obscure or distort their satellite view. Inaccuracies in the cloud proper- ties are caused by mixing clouds that feed from the boundary layer with clouds that have elevated bases above the bound- ary layer, and hence are not interacting with the aerosols there. Another yet unresolved challenge is correct retrieval of the properties of ensembles of convective clouds. A seri- ous limitation is the inability of the aerosols and clouds in conditions of high fractional cloud coverage to be retrieved simulataneously. As a result, the practice of relating retrieved cloud to retrieved aerosol properties is plagued by large er- rors in both parameters, which may cause both random errors and systematic biases. Mathematically, adding random noise to two physically linearly related variables (e.g., Y = aX) causes the correlation to decrease and the slope to tend to- ward zero. Therefore, the large observational errors in the de- pendence of clouds on aerosol properties have likely caused an underestimate of the slopes of their relationships, which is deﬁned to be the observed aerosol indirect effect (AIE). This error might explain the low estimates of the observed AIE as compared to results of model simulations (e.g., Quass et al., 2009; Penner et al., 2011). Table 1. The spectral bands of the Multi-Spectral Imager, which will have a footprint of 100 m. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Wave band Channel name SNRa or NEDTb 450–470 nm Blue SNR = 4 560–580 nm Green SNR = 4 670–690 nm Red SNR = 4 1070–1200 nm Vapor wide SNR = 5 1120–1150 nm Vapor narrow SNR = 5 1365-1395 nm Cirrus absorption SNR = 2 2080–2150 nm 2.1 Phase water SNR = 2 2200–2310 nm 2.3 Phase ice SNR = 2 3420–3950 nm 3.7 Effective radius SNR = 5; NEDT = 0.2 K at 246 K 8.3–9.25 µm 8.7 Temperature NEdT = 0.1 K at 232 K 10.2–11.2 µm 10.7 Temperature NEdT = 0.1 K at 270 K 11.2–12.3 µm 11.7 Temperature NEdT = 0.2 K at 300 K a SNR, signal-to-noise ratio. Required SNR values calculated for 1 % albedo and 30◦ solar and viewing angles. b NEDT, noise equivalent differential temperature. Cloud vertical motions will be obtained by using a se- quence of stereoscopic images that map the evolution of the cloud surface. The Multi Angle Imager (MAI) onboard CHASER has this capability. The MAI consists of three visi- ble high resolution cameras. The center camera points 30◦off nadir eastward. The other two cameras point 30◦off nadir across-track (eastward) and ±30◦off nadir along-track to achieve a stereo geometry with the Sun at the back as illus- trated in Fig. 1. The ﬁeld of view of each camera is 50 m by 102.4 km, with a spatial resolution of 50 m at their ground center track. The three cameras cross scan the same ground track, providing a sequence of three images 1 minute apart, from three different viewing angles. The idea of vertical mi- crophysical proﬁling from space was introduced by Rosen- feld and Lensky (1998), using the Advanced Very High Res- olution Radiometer (AVHRR) observing the temperature and particle effective radius (re) at the tops of convective clouds at various stages of vertical development. This idea was im- proved to a proposed side scanning satellite (Martins et al., 2011), and further developed by enhancing the resolution and with the addition of the MAI for CHASER (Renno et al., 2012). 1.3 Uncertainties in global measurements of cloud-mediated radiative forcing by aerosols Based on this brief summary of aerosol effects on clouds, it is evident that having accurate measurements of CCN acti- vation spectra as a function of water vapor supersaturation – CCN(S) – is vital for a quantitative understanding of the weather systems and the aerosol impacts on the Earth energy budget and climate. This information is a necessary compo- nent for disentangling the effects of aerosols from those of the greenhouse gases on the Earth energy budget, and hence for understanding climate change and for climate prediction. – CCN(S) – is vital for a quantitative understanding of the weather systems and the aerosol impacts on the Earth energy budget and climate. This information is a necessary compo- nent for disentangling the effects of aerosols from those of the greenhouse gases on the Earth energy budget, and hence for understanding climate change and for climate prediction. Global measurements are practical only from space. Aerosol properties such as aerosol optical depth have been measured from space, but the interpretation of the measur- able quantities into CCN(S) is still challenging. The radiative forcing caused by impacts of aerosol on clouds is inferred Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 1. An illustration of the measured vertical proﬁles of cloud pa- rameters pointing 30◦off track toward sun-illuminated cloud sur- faces. Fig. 1. An illustration of the measured vertical proﬁles of cloud pa- rameters pointing 30◦off track toward sun-illuminated cloud sur- faces. physical entities determining Na, which in turn determine the remaining cloud properties (e.g., cloud drop effective radius and initiation of rain as a function of height above base). Re- trieving Na and cloud base updraft velocities is required for disentangling the impacts of dynamics from microphysics on the various cloud properties. The concept of using ground-based cloud observations for retrieving CCN was originally proposed by Feingold et al. (1998). They retrieved CCN using measurements of wb with vertically pointing Doppler cloud radar, and Nd esti- mated from a combination of cloud radar and passive mi- crowave measurements. Obtaining the size distribution of the aerosols by lidar or other measurements can provide the CCN supersaturation activation spectrum. This method was applied to ground-based measurements of marine stratocu- mulus clouds at Point Reyes, California, to obtain the aerosol effects on cloud radiative forcing (McCominsky and Fein- gold, 2011). They showed that the variability in liquid water path (LWP) that occurs due to 3-D effects and cloud mixing tends to reduce the measured strength of the albedo effect. Examples from aircraft measurements (from Freud et al., 2011) are shown in Figs. 2–4. Another fortunate fact is the highly repeatable tight linear relationship between rv and cloud droplet effective radius, re (see Fig. 5), a parameter that can be measured by satellites (Nakajima and King, 1990). Freud et al. (2011) explain this methodology in more detail. The proposed methodology represents a signiﬁcant ad- vancement, as it can use satellite measurements and therefore provide global coverage for retrieved CCN and cloud proper- ties. Furthermore, variability in LWP for a cloud with a given depth and Nd is not a source of error for this methodology. Based on this physical description, Na is the most funda- mental microphysical property of a convective cloud. It de- termines the rate at which the droplets grow with cloud depth in the updraft and in turn the rate at which they are con- verted into precipitation-sized particles. Na also affects the radiative properties of the clouds, as higher concentrations reduce the droplet sizes for a given amount of cloud wa- ter (Twomey, 1977). D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2042 The latter is determined by CCN(S) and the updraft speed at cloud base. In a hypothetical cloud parcel rising adiabati- cally, the diameters of the cloud droplets grow in proportion to the cube root of the adiabatic cloud liquid water content [mixing ratio], qLa. This occurs because qLa = 4/3 ρLπr3 vNa, where rv is the mean cloud droplet volume radius, and ρL is the water density. Na can be readily calculated because qLa is a known thermodynamic function of the cloud depth, D (the vertical distance above cloud base). Real clouds are far from being adiabatic, of course, but fortunately the mixing and dilution of convective clouds occurs in a way that leaves the rv as if the clouds were nearly adiabatic. This occurs because clouds mix nearly inhomogeneously (e.g., Paluch and Baumgardner, 1989). That is, cloud droplets directly ex- posed to mixing evaporate completely leaving the remaining droplets unaffected. This leads to a reduction of Nd and of the cloud water content, qL, but mixing does not affect rv. Bur- net and Brenguier (2007), based on aircraft measurements with a sampling rate of 10 Hz (providing 10-m resolution in cloud), reported a state that is more intermediate between the two extreme mixing states, whereas Paluch and Baumgard- ner (1989) and Freud et al. (2011) using a sampling rate of 1 Hz, or a scale of 100 path length in clouds. According to Lehmann et al. (2009) mixing is expected to become more homogeneous at smaller scales. This apparent disagreement can be resolved when realizing that mixing goes to the ho- mogeneous at the very small scales (<10 m). Taking it to the limit, at the scale of the single droplet the mixing by deﬁ- nition is homogeneous, as the droplet must evaporate gradu- ally and not vanish instantaneously when exposed to dry air. When observing larger volumes, the time or space scale at which the droplets evaporate becomes smaller with respect to the observation scale. At the extreme, there is a clear de- marcation between cloud and cloud free air, as water clouds do not diffuse gradually into their environment as smoke or cirrus clouds do. The satellite views the clouds at the 100 m scale, which is similar to the aircraft measurement scale at 1 Hz being ∼100 m ﬂight path. Therefore, the 100 m scale is the relevant one for the application to CHASER. Fig. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Na reﬂects not only CCN(S), but also the updraft velocities, as manifested in the actual supersat- uration that these CCN were exposed to. However, direct 2 A new proposed concept for measuring aerosol indirect effect This paper introduces a new concept for measuring simulta- neously CCN(S) and the microphysical and dynamical prop- erties of the clouds for assessing quantitatively the cloud- interactions with global coverage from satellites. A proposed satellite mission named “Clouds, Hazards, and Aerosols Sur- vey for Earth Researchers” (CHASER) is based on this con- cept (Renno et al., 2012). The satellite has two main instru- ments: the Multi-Spectral Imager (MSI) that will have a ﬁeld of view of 100 m by 102.4 km at the center of the ground track for all wavelengths. The satellite view will point 30◦ off nadir eastward, with the Sun at 30◦off zenith on the west, at a local solar time of 14 LST. The MSI will have 12 spec- tral bands in the visible, near infared (IR) and thermal IR, as shown in Table 1. Traditionally, CCN are measured in laboratory or ﬁeld ex- periments with a cloud chamber where known water vapor supersaturation (S) is applied to an air sample and the ac- tivated cloud droplets are counted. Here, we view clouds as natural cloud chambers where the number of nucleated aerosols in cloud droplets, Na, and S can be retrieved, and hence the CCN spectrum can be obtained. Speciﬁcally, the approach uses the retrieved Na from the dependence of the cloud drop size on the potential amount of condensed cloud water as a function of distance above cloud base, along with retrieved updrafts near cloud base, wb, for calculating the CCN activation spectra as a function of S. This information is used to map CCN in the cloudy boundary layer globally. This is important because this maps CCN and wb, the fundamental www.atmos-meas-tech.net/5/2039/2012/ www.atmos-meas-tech.net/5/2039/2012/ Atmos. Meas. Tech., 5, 2039–2055, 2012 D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2043 Fig. 2. Aircraft observations of tight relationships between re and altitude in a cluster of deep convective clouds over India. Each red circle marks the re measured in a 1-s (∼100 m) ﬂight path. The dark curve shows the proﬁle of the adiabatic re (rea). The ﬁgure is taken from Freud et al. (2011), The Na is calculated by the method- ology described in Freud et al. (2011). It can be seen that for any given cloud depth the scatter of re is fairly small, and that values are normally within 2 µm from rea, despite including measurements with down to 5 % of adiabatic water. The blue circles indicate the expected effective radii in case of fully homogeneous mixing with entrained air which has 50 % relative humidity (RH), which was ob- tained from a nearby sounding. They are calculated for the mixing proportions of the actually measured adiabatic fractions. Similar re- lations are shown elsewhere in the world by Rosenfeld and Freud (2012). Fig. 2. Aircraft observations of tight relationships between re and altitude in a cluster of deep convective clouds over India. Each red circle marks the re measured in a 1-s (∼100 m) ﬂight path. The dark curve shows the proﬁle of the adiabatic re (rea). The ﬁgure is taken from Freud et al. (2011), The Na is calculated by the method- ology described in Freud et al. (2011). It can be seen that for any given cloud depth the scatter of re is fairly small, and that values are normally within 2 µm from rea, despite including measurements Fig. 3. Relationship between re and D for various values of Na in cm−3. The value derived from the proﬁle of re measured by the aircraft is Na = 610 cm−3 (580 mg−1), within 5 % of the directly aircraft-measured maximum cloud base droplet concentration. Fig. 3. Relationship between re and D for various values of Na in cm−3. The value derived from the proﬁle of re measured by the aircraft is Na = 610 cm−3 (580 mg−1), within 5 % of the directly aircraft-measured maximum cloud base droplet concentration. Fig. 2. Aircraft observations of tight relationships between re and altitude in a cluster of deep convective clouds over India. Each red circle marks the re measured in a 1-s (∼100 m) ﬂight path. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations The dark curve shows the proﬁle of the adiabatic re (rea). The ﬁgure is taken from Freud et al. (2011), The Na is calculated by the method- ology described in Freud et al. (2011). It can be seen that for any given cloud depth the scatter of re is fairly small, and that values are normally within 2 µm from rea, despite including measurements with down to 5 % of adiabatic water. The blue circles indicate the expected effective radii in case of fully homogeneous mixing with entrained air which has 50 % relative humidity (RH), which was ob- tained from a nearby sounding. They are calculated for the mixing proportions of the actually measured adiabatic fractions. Similar re- lations are shown elsewhere in the world by Rosenfeld and Freud (2012). Fig. 3. Relationship between re and D for various values of Na in cm−3. The value derived from the proﬁle of re measured by the aircraft is Na = 610 cm−3 (580 mg−1), within 5 % of the directly aircraft-measured maximum cloud base droplet concentration. the extra time and potential spectrum broadening due to typ- ically weaker updrafts and preferred (partial) evaporation of the smaller droplets upon mixing in drier air. Additionally, the rate of droplet coalescence is proportional to ∼r5 e (Freud and Rosenfeld, 2012), which practically implies the exis- tence of a threshold re above which efﬁcient warm rain for- mation can occur. This threshold is reached at a known height above cloud base with a given Na, because the vertical proﬁle of re closely follows the theoretical adiabatic condensational growth curve, even in diluted clouds. The small observed de- viations are mainly caused by deviations from purely inho- mogeneous mixing which causes partial droplet evaporation. Consequently, the minimal cloud depth for precipitation for- mation (Dp) must theoretically change nearly linearly with Na because the adiabatic water content increases nearly lin- early with Dp, with a slope that depends on the cloud base pa- rameters. This means that in highly polluted clouds or where strong cloud-base updrafts occur (i.e., large Na), clouds may need to grow well above the freezing level, even in the trop- ical atmosphere, before precipitation forms either by warm or by mixed-phase processes. For full detail see Freud and Rosenfeld (2012). D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations measurement of Na is usually not possible because entrain- ment of sub-saturated ambient air into the cloud decreases the cloud droplet concentrations by evaporation and dilu- tion. Even the cores of deep convective clouds, where aircraft measurements are normally avoided due to the strong verti- cal motions and icing hazards, are prone to entrainment. This is mainly because of the fairly small horizontal extent and the strong turbulence in and near the convective cloud towers. 4 Na controls the height for the onset of warm rain The droplet condensational growth is determined by the number of activated CCN (Na) and the height above cloud base, D. When the droplet effective radius (re) exceeds ∼14 µm, considerable precipitation mass is likely to form in growing convective clouds regardless of the extent of entrain- ment and mixing. This is because the low droplet concen- trations and liquid water content in the diluted parts of the cloud, which slow the coalescence rate, are compensated by Atmos. Meas. Tech., 5, 2039–2055, 2012 3 Na as a fundamental extensive cloud property In this section we show that the number of activated aerosols at cloud base, Na, is a fundamental property that determines to a large extent the microstructure of the cloud as a whole. Na is determined by the aerosol size distribution, chemical composition, and water vapor supersaturation at cloud base. Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations www.atmos-meas-tech.net/5/2039/2012/ Nd (cm-3) re (m) (cm-3) Clo tion, Nd ( oud drop e oncentrat effective ud drop co radius, re Clou e (m) Time 0 5 10 15 20 0 5 10 15 20 India Israel California Texas y = 1.08x R= 0.993 y = 1.08x R= 0.991 y = 1.08x R= 0.989 y = 1.08x R= 0.995 Effective Radius [µm] Mean Volume Radius [µm] Mean Volume Radius [µm] Fig. 4. The cloud drop concentration, Nd, and effective radius, re, at a horizontal cross section through a convective tower. Note the relative stability of re up to the cloud edge while Nd is highly vari- able. This is a manifestation of the nearly inhomogeneous mixing that dilutes Nd while keeping re nearly constant. Fig. 5. The cloud droplet effective radius, re, versus the droplet mean volume radius, rv, for 1 Hz averaged droplet size distributions measured for various locations, cloud types and by different cloud droplet probes. Note the universally highly repeatable and tight re– rv relationship (from Freud and Rosenfeld, 2012). Fig. 5. The cloud droplet effective radius, re, versus the droplet mean volume radius, rv, for 1 Hz averaged droplet size distributions measured for various locations, cloud types and by different cloud droplet probes. Note the universally highly repeatable and tight re– rv relationship (from Freud and Rosenfeld, 2012). www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2044 feld etal.: Satellite mission to retrieve CCN concentrations 0 5 10 15 20 0 5 10 15 20 India Israel California Texas y = 1.08x R= 0.993 y = 1.08x R= 0.991 y = 1.08x R= 0.989 y = 1.08x R= 0.995 Effective Radius [µm] Mean Volume Radius [µm] Fig. 5. The cloud droplet effective radius, re, versus the droplet mean volume radius, rv, for 1 Hz averaged droplet size distributions measured for various locations, cloud types and by different cloud droplet probes. Note the universally highly repeatable and tight re– rv relationship (from Freud and Rosenfeld, 2012). 2044 D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Nd (cm-3) re (m) (cm-3) Clo tion, Nd ( oud drop e oncentrat effective ud drop co radius, re Clou e (m) Time Fig. 4. The cloud drop concentration, Nd, and effective radius, re, at a horizontal cross section through a convective tower. Note the relative stability of re up to the cloud edge while Nd is highly vari- able. This is a manifestation of the nearly inhomogeneous mixing that dilutes Nd while keeping re nearly constant. 0 5 10 15 20 0 5 10 15 20 India Israel California Texas y = 1.08x R= 0.993 y = 1.08x R= 0.991 y = 1.08x R= 0.989 y = 1.08x R= 0.995 Effective Radius [µm] Mean Volume Radius [µm] Fig. 5. The cloud droplet effective radius, re, versus the droplet mean volume radius, rv, for 1 Hz averaged droplet size distributions measured for various locations, cloud types and by different cloud droplet probes. Note the universally highly repeatable and tight re– rv relationship (from Freud and Rosenfeld, 2012). Nd (cm-3) re (m) (cm-3) Clo tion, Nd ( oud drop e oncentrat effective ud drop co radius, re Clou e (m) Time Fig. 4. The cloud drop concentration, Nd, and effective radius, re, at a horizontal cross section through a convective tower. Note the relative stability of re up to the cloud edge while Nd is highly vari- able. This is a manifestation of the nearly inhomogeneous mixing that dilutes Nd while keeping re nearly constant. 5 Retrieving Nd from satellites observations of clouds can be used for retrieving Na and its precursors, CCN(S) and wb. Satellite retrieval of the column-integrated number of cloud droplets [cm−2] is based on the retrieved vertical liquid wa- ter path (LWP). When LWP is given, the vertically averaged cloud droplet number density Nd can be estimated. Indirect measurements of Na by satellite and lidar retrievals were previously applied to shallow marine stratiform clouds, with the main assumption that the clouds are composed of nearly adiabatic elements (Bennartz, 2007; Brenguier et al., 2000; Schuller et al., 2003; Snider et al., 2010). These retrievals had large uncertainties mainly due to violation of the adiabatic assumption and were not always validated with direct mea- surements. The retrieval of Nd worked well in several cases of extreme homogeneous marine stratocumulus, where these assumptions are closest to reality (Painemal and Zuidema, 2011). Furthermore, this methodology is not applicable to convective clouds due to their larger departure from the as- sumption that they are close to adiabatic as compared to ma- rine startocumulus, and also due to the variable cloud top heights and depths at scales smaller than the typical satellite sensor resolution of >1 km. www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 1. Retrieve re(D), the vertical proﬁle of re, as a function of vertical distance above cloud base. 1. Retrieve re(D), the vertical proﬁle of re, as a function of vertical distance above cloud base. rain forms, then the unique relation between the vertical pro- ﬁle of re and Na weakens. Therefore, the derivation of Na is expected to be most accurate for re values smaller than ∼14 µm, where coalescence starts to dominate the cloud drop size distribution (Freud and Rosenfeld, 2012; Rosenfeld et al., 2012c). Errors in the measurements of re and departures from the extreme inhomogeneous assumption would affect the determination of Na, and therefore the effects of mixing would need to be accounted for. rain forms, then the unique relation between the vertical pro- ﬁle of re and Na weakens. Therefore, the derivation of Na is expected to be most accurate for re values smaller than ∼14 µm, where coalescence starts to dominate the cloud drop size distribution (Freud and Rosenfeld, 2012; Rosenfeld et al., 2012c). Errors in the measurements of re and departures from the extreme inhomogeneous assumption would affect the determination of Na, and therefore the effects of mixing would need to be accounted for. 2. Convert the satellite retrieved re to rv using the very tight and invariable linear relations between the two quantities, as shown in Fig. 2 here and more fully in Fig. 5 of Freud and Rosenfeld (2011). 3. Invoke the assumption of inhomogeneous mixing, which in the extreme condition means that rv equals rva, the adiabatic rv. This allows retrieving Na in the follow- ing way: Accurate measurements of re require vertical resolutions of the order of 100 m. Moreover, accurate measurements of re as a function of the vertical distance above cloud base, D, require the minimization of 3-dimensional effects. The reduction of this effect can be accomplished by making mea- surements of re at wavelengths of about 3.7 µm, because the strong absorption of radiation by water at this wavelength reduces 3-dimensional effects (Rosenfeld et al., 2004; Paine- mal and Zuidema (2011). At shorter wavelengths where ab- sorption is weaker (1.6 and 2.1 µm), a larger fraction of the light exiting a measured cloud pixel has been scattered by cloud particles inside and outside of the intended cloud measurement volume, thus degrading the accuracy of re- trieved re. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Therefore, measurements at 3.7 µm are inherently more accurate for retrieving Na than measurements at 1.6 or 2.1 µm. Na = 3/(4qLaρLπr3 va). (1) (1) 4. Correct Na for the extent of homogeneous mixing and the resultant reduction of rv with respect to rva. This correction is done based on the relative humidity near the cloud. This procedure will be tuned by analysis of already available aircraft measurements (Freud et al., 2011). Instead of correcting for the extent of homogeneous mix- ing (step 4 above), it can be assumed that the highest re val- ues from each level in the cloud (there will be a range of re values for each level because of the high resolution of the re- trievals) represent the least diluted cloud elements, and there- fore are closest to the adiabatic re. Therefore, Eq. (1) may be used for deriving Na (after replacing re with rv) from the ﬁt- ting of the best re proﬁle to the highest re values at each level. This proﬁle would resemble the enveloping curve shown in Fig. 2 (dark grey line), which was calculated for an assumed Na. Charts like the one shown in Fig. 3 can be used for ob- taining Na when re(D) is retrieved. This way for deriving Na does not require accounting for the mixing inhomogeneity, so it is quite simple and straight-forward. On the other hand it utilizes only a relatively small number of re values from each proﬁle, and therefore may be more sensitive to noise in the data; the resultant Na would be biased in the case that the utilized re values would be from cloud elements far from adiabatic. Taking into account the deviation from the extreme inho- mogeneous mixing scenario when deriving Na from the ver- tical proﬁle of re is not straightforward. Freud et al. (2011) showed that with the inhomogeneous mixing assumption Na was over-estimated on average by ∼30 %. This is because the measured re needs to be extrapolated to its adiabatic value (and converted to rv) before Eq. (1) can be applied. How much to add to the measured re depends mainly on the amount of entrainment and the relative humidity (RH) of the entrained air. The RH may be estimated from the measure- ments of the absorption of the water vapor (in the 1120– 1150 nm band) near the edges of the clouds. 6 The scientiﬁc basis for retrieving Na Recent studies based on airborne measurements in deep con- vective clouds show that the cloud droplet effective radius (re) anywhere in the cloud is a strong function of the num- ber of activated CCN (Na) and the distance above cloud base (D) (Pawlowska et al., 2000; Andreae et al., 2004; Freud et al., 2008 and 2011; Freud and Rosenfeld, 2012; Rosenfeld et al., 2008b). The entrainment of sub-saturated air from the surroundings of the cloud and the mixing with the cloudy air, although reducing the cloud droplet concentrations and the liquid water content by evaporation and dilution, appar- ently does not have a strong effect on re. The common ex- planation is that the mixing is strongly inhomogeneous, i.e., that the cloud droplets at the edges of a blob of entrained sub-saturated air quickly and completely evaporate while in- creasing the vapor content in the entrained air until it is sat- urated and cannot evaporate additional droplets. This lowers the droplet concentrations but leaves the shape of the droplet size spectra unchanged and hence does not affect re. This is why if extreme inhomogeneous mixing is assumed and the cloud base properties are known, Na can be derived from the vertical proﬁle of re as long as the droplets grow mainly by condensation. When droplet coalescence is active and warm The fact that Nd is vertically averaged and is subject to an unknown extent of dilution with ambient air renders it to be related only loosely to CCN. In contrast, Na is the fundamental property representing the combined effects of CCN(S) and cloud base updraft velocity, wb, which in turn determine other cloud properties. Consequently, the objec- tive of this study is to introduce a method by which satellite Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ 2045 www.atmos-meas-tech.net/5/2039/2012/ Atmos. Meas. Tech., 5, 2039–2055, 2012 7 The scientiﬁc basis for obtaining the updraft velocity near cloud base Cloud vertical motions can be obtained in principle by us- ing a sequence of stereoscopic images that map the evolu- tion of the cloud surface. The Multi Angle Imager (MAI) on- board CHASER has this capability. Two stereoscopic views 1 min apart can in principle provide the vertical component of the cloud surface evolution, by tracking protuberances at the cloud envelope. Tracking such protuberances near cloud base with an oblique view from the side can provide the ver- tical motion of these features, which approximates the mean updraft at cloud base. An illustration of such cloud features is given in Fig. 6. The challenge here is separating the observed movement into horizontal and vertical components. This can be assisted by the independent knowledge of the height for the central view of the cloud surface. This height can be ob- tained from the cloud temperature as measured by the MSI that is aligned with the center camera of the MAI. The cloud surface temperature can be uniquely related to cloud surface height in convective clouds, when using the moist adiabatic lapse rate and the cloud base temperature. Systematic bias errors in the height–temperature relations in the same cloud cluster would not affect the accuracy of the retrieval, and the vertical lapse rate is rather constant, as determined by known thermodynamic factors that do not vary much within the do- main. The accuracy of the height retrieval by two Multi-angle Imaging SpectroRadiometer (MISR) cameras is about 300– 500 m (Hovarth and Davis, 2001; Moroney et al., 2002). The MISR has a horizontal nadir resolution of 275 m, versus the 50 m footprint of the MAI. This alone should reduce the error by a factor of 5, resulting in an accuracy of 60–100 m. The off nadir tilt of the MAI viewing geometry and the height ref- erence provide additional signiﬁcant improvement in the ac- curacy. This addresses the main source of inaccuracy, which is the uncertainty with respect to the separation of the ob- served cloud motion to vertical and horizontal components. The error of the motion of perfectly tracked cloud features is determined by uncertainty between the projections of the vertical and horizontal components of the cloud motion on the viewing angle from the satellite. Fixing one of the com- ponents allows the unambiguous determination of the other. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations It is proposed that the vertical proﬁle of vapor near clouds will be retrieved by the derivative of the precipitable water as a function of height, when measured above the cloudy pixels at different heights in the cloud cluster of interest. The relative humidity will be calculated based on the known temperature as a func- tion of height, as obtained from the measured temperatures of the reﬂecting reference cloud tops at the various heights. The accuracy of this method has not yet been determined, but it is expected to be much more accurate than the alterna- tives of forecast/reanalysis or derivation by satellite inversion techniques (e.g., Singh and Bhatia, 2006 and the references therein), because in this method the clouds serve as reﬂectors at known heights within the range of interest. Furthermore, the ability to calculate many vertical proﬁles for the same cloud around its sunlit side allows quantifying the local gra- dients and inhomogeneity of the moisture ﬁeld. The increase in re with cloud depth during the droplet con- densational growth depends on Na and the mixing ratio of the water vapor at cloud base. When re reaches ∼14 µm in the convective clouds, warm rain starts to form (e.g., Rosenfeld, 1999 and 2000; Rosenfeld and Gutman, 1994; Gerber, 1996). As a result, if the cloud base temperature and pressure are known, the cloud depth at which re reaches the 14 µm warm rain threshold (Dp) depends only on Na. However, since the mixing ratio of the adiabatic water increases nearly linearly with cloud depth, Na and Dp should be linearly related for constant cloud base properties. Freud and Rosenfeld (2012) demonstrated this simple relationship with a small number of deep convective clouds over Israel and India. It would be interesting to see whether this relationship between Dp and Na holds for a larger number of cases in different parts of the g y Here, we retrieve Na using the following algorithm: www.atmos-meas-tech.net/5/2039/2012/ Atmos. Meas. Tech., 5, 2039–2055, 2012 2046 D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations world based on the proposed methodology to derive Na from space. If it holds, then it would be very useful to include this simple parameterization in coarse-resolution models that do not resolve single clouds. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations The objective retrieval of CCN requires the ability to re- trieve re accurately for the smallest possible fair weather boundary layer clouds and still resolve their vertical mi- crostructure. The re is retrieved based on the observed ex- tent of absorption of solar radiation in the clouds. Because 3.7 µm radiation is absorbed in water 10 times more strongly than 2.2 µm radiation, clouds are more opaque at 3.7 µm and hence less affected by light arriving from outside of the mea- suring volume. The smallest practical size that can be used with little interference from adjacent cloud volume at 3.7 µm is about 100 m (Rosenfeld et al., 2004). Taking into account the 3-D effects that are inherent to such small convective clouds allows the retrieval of re in convective clouds to an accuracy of 1 µm (Zinner et al., 2008). An error of a factor e in the retrieved re would be ampliﬁed by e3 in the calcu- lated Na. This means that an overestimate error of 1 µm for a cloud with re=15 µm would propagate to a bias error in Na of (16/15)3 = 1.21. This is a manageable uncertainty. Fig. 6. Illustration of the topography of three small convective cloud segments. The contours show the possible development lines of the cloud top along three time steps, i.e., three views of the same cloud at t1, t2 and t3. These identiﬁable cloud features just above cloud base develop vertically at a similar rate as the cloud base updraft. The MAI 50 m resolution is illustrated at the scale bar. The resolu- tion appears to be sufﬁcient for mapping the marked perturbations and track these features along a sequence of three 1-min images, and map the average topographies for t1 to t2 and for t2 to t3. The differentiation of the two topographic maps will provide the cloud vertical motion. It appears that the retrieved updraft near cloud base, wb, would be the largest for cloud 1 and the smallest for cloud 3. www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations State-of-the-art in situ measurements are based on size selection of aerosol parti- cles followed by their activation to CCN in a cloud chamber, so that the dependence of activation on size and composition can be studied separately (Rose et al., 2008; Su et al., 2010). In order to separate the vertical and horizontal compo- nents, it is necessary to assume something as being ﬁxed. In the situation of a well-mixed boundary layer from which convective clouds develop, the base of the clouds is ﬂat at the convective condensation level, as determined by thermody- namic considerations. This height varies at a negligible rate compared to the vertical motions within the clouds. There- fore, cloud surface elements that are close to the base can be assumed to have the same horizontal motion as the cloud base. Under such an assumption, all the relative angular dis- placement of the cloud surface with respect to the cloud base can be ascribed to the vertical dimension. The mean horizon- tal wind has to be taken into account from the average motion of cloud bases. These data are most beneﬁcial for improving our under- standing of how aerosols affect climate if they are used to test and improve the prognostic aerosol models that are incor- porated into climate models. For this purpose, the CCN(S) spectra obtained from the CHASER measurements of the concentration of activated CCN as a function of the water vapor supersaturation, Na (S), are considered to represent the CCN concentrations as a function of particle size (d) and composition (Reutter et al., 2009). Figure 7 illustrates the re- lation between S and Na calculated as a function of the up- draft speed near the cloud base (wb), which can be described by Eq. (2): In more general terms, the base of convective clouds is ﬂat and at constant height. This provides a ﬁxed reference for the MAI’s three views of a single cloud and allows the horizontal motion at cloud base to be determined. The knowledge of the horizontal motion at the cloud base allows the separation of the horizontal and vertical components of the motion of cloud protuberances measured by MAI. The accuracy of the vertical speed retrieval is approx- imately equal to the vertical motion of one pixel of 1z = 50 m between measurements 1t = 60 s apart, resulting in 1z/1t = 0.8 m s−1. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations spectra could be represented by a CCN concentration nor- malized to S = 1 %, and an exponent b. More recent studies have moved away from this approach for two reasons. First, it was found that the power law was often a poor approximation for the observed CCN spectra, and therefore, b values could not be reliably associated with speciﬁc aerosol types (Gunthe et al., 2009; Rose et al., 2010). The second and more impor- tant reason is that this representation treats CCN as a “black box” by using a parameterization that does not contain any fundamental physical properties, and thus it is not amenable to prognostic modeling. We must have cloud base height and temperature for making a retrieval. Clouds have to be some distance apart in order to see the entire sides down to the cloud base. This distance can be computed by a simple geometrical calculation. One km deep clouds have to be at least 600 m apart for the retrieval to be possible. The cloud base and a piece of ground as its background, for a reference to the cloud base, needs to be observed. The calculation of the supersaturation of the water vapor pressure near cloud base requires the determination of the updraft speed at cloud base (wb). It is assumed that the verti- cal speed of cloud protuberances near cloud base is approx- imately equal to the updraft speed. The challenge is to sepa- rate the horizontal and the vertical components of the motion. This is accomplished by the procedure described next. Current CCN research shows that the activation of an aerosol particle at a speciﬁc S is predominantly determined by the number of water-soluble molecules or ions that it contains. The number of soluble molecules or ions, in turn, depends on the aerosol volume and composition (the num- ber of soluble molecules per unit volume). The volume of the aerosol particle is speciﬁed by its dry diameter while the composition is speciﬁed by the hygroscopicity param- eter, κ (Petters and Kreidenweis, 2007). In this approach, the CCN properties of aerosols can be related directly to their basic physical and chemical properties, and therefore are amenable to prognostic modeling. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Tracking at least 10 pixels of a sin- gle protuberance increases the accuracy of the calculation to ∼0.2 m s−1. This provides measurements with the neces- sary accuracy for calculating S even in marine stratocumu- lus clouds. However, a more cautious estimate of 0.5 m s−1 would still provide useful data for marine stratocumulus and will certainly sufﬁce for other kinds of convective clouds. S =3.78 w0.64 b N−0.4 a . (2) (2) These calculations indicate that overestimates of 20 % in Na cause S to be underestimated by 7 %, whereas overestimates of 20 % in wb cause S to be overestimated by 12 %. Since the error of the estimates of Na and wb is ∼10 % to 20 %, the error in the calculation of S is ∼10 %. These calculations indicate that overestimates of 20 % in Na cause S to be underestimated by 7 %, whereas overestimates of 20 % in wb cause S to be overestimated by 12 %. Since the error of the estimates of Na and wb is ∼10 % to 20 %, the error in the calculation of S is ∼10 %. Since aerosol particle size and composition affect CCN activation and can vary independently of one another, there is no unique way to convert a CCN(S) spectrum into a CCN(d, S) spectrum that would allow the retrieval of the desired functions Na(d) and κ(d). That is, the particle num- ber concentration and hygroscopicity cannot be retrieved as a function of size without additional assumptions. Fortu- nately, recent research has yielded considerable information about κ from both theoretical studies and laboratory and ﬁeld 7 The scientiﬁc basis for obtaining the updraft velocity near cloud base This means that using a ﬁxed reference height of a ﬂat cloud base at the top of a well-mixed boundary layer for all three views can further improve substantially the accuracy with re- spect to the calculation above by at least a factor of about two, leading to an accuracy of about 50 m. Viewing the cloud at an angle of 30 degrees from the zenith allows the cloud bases to be measured, unless they are tilted towards the satellite by an angle that approaches 30 degrees. The tilt angle can be identiﬁed by the multi- angle views. The determination of the cloud base in the cen- tral view is assisted by the cloud temperature. The warmest cloudy pixels represent temperature just above cloud base. The cloud base updraft is highly correlated to the cloud top rate of rising above it (Blyth et al., 2005). Aircraft measure- ments show that the cloud top rising rate is on average half of the peak updraft within the cloud below the tops. Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ 2047 Atmos. Meas. Tech., 5, 2039–2055, 2012 9.1 Retrieving errors in cloud drop effective radius A comparison with observations shows that modeled and observed κ agree to ≤0.05 for 10 out of 14 stations. Field campaigns have also shown that κ does not usually change strongly with size over the range relevant to CCN active in convective clouds (0.05–0.5 µm). The accuracy of retrieval of water vapor above clouds by differential absorption near 0.9 µm was shown to be about 0.2 mm (Albert et al., 2001). The accuracy is expected to be much improved at the 1.2 µm waveband due to the greater absorption of vapor at that range. p p g An overestimate error of 0.2 mm in the precipitable wa- ter was calculated to incur an underestimate error of about 0.5 µm in re for re = 14 µm. The error decreases for smaller re. This is a rather large sensitivity. The retrieval of re at 2.1 µm is free of errors incurred by temperature measure- ments and has 1/3 of the 3.7 µm error due to inaccuracies in precipitable water above cloud. While the retrieved errors of re at 2.2 µm are generally higher due to the smaller de- pendence on intervening vapor and the lack of need to sep- arate the solar and thermal components of the radiation, the greater absorption at 3.7 µm (Mitchel, 2002) minimize the 3- dimensional effects in the convective cloud elements, thus improving the accuracy for small cloud elements and for small distances above cloud base. The solar reﬂectance of the same opaque cloud in the visible is calculated to be at 3.7 µm half of the reﬂectance at 2.1 µm . The retrieved re at 3.7 µm of the smallest resolvable cloud depth of 100 m with re of 7 µm, Once κ is known or assumed, the distribution CCN(S) can be inverted to a distribution CCN(d) because for each κ there is a functional relationship between the supersatura- tion and the critical particle diameter above which particles can be activated to cloud droplets. Thus, each supersatura- tion step in the CCN(S) spectrum corresponds to a size step in the CCN(d) spectrum, and the difference in CCN num- ber concentration between the supersaturation steps yields the number of CCN in the corresponding size interval (Su et al., 2010). 9.1 Retrieving errors in cloud drop effective radius The sensitivity of the retrievals of re was tested for this study, using the MODIS Airborne Simulator aircraft data near the southern tip of Florida on 28 July 2006, at a sensor zenith an- gle around 30◦eastward of nadir, and solar zenith angle 35◦ westward of zenith. The surface footprint at this geometry was near 70 m. An error of 1◦K overestimate in brightness temperature of 3.7 µm incurs an error of 0.8 µm underestimate in re for re = 14 µm. According to Table 1, an instrument measure- ment noise of NEDT of 0.2◦K in the 3.7 µm channel would translate to added noise in re of 0.16 µm. Fig. 7. The CCN spectra can be calculated using measurements or model simulations of Na and S(wb). This chart was drawn using data from P¨oschl et al. (2010) for the activation of organic aerosol particles with κ = 0.13 as observed for pristine tropical rainforest air. κ = 0.3 is more representative for most other continental aerosol regimes. Increasing κ would lower the S values in the ﬁgure (Reut- ter et al., 2009). The error becomes larger at greater re and less at smaller re. An error of 1◦K overestimate in brightness temperature of 10.7 µm incurs an error of 0.4 µm overestimate in re for re = 14 µm. The error becomes slightly larger at smaller re. According to Table 1, an instrument measurement noise of NEDT of 0.1◦K in the 10.7 µm channel would translate to added noise in re of 0.04 µm. An error of a factor of 1.1 overestimating the precipitable water above low cloud tops at 20◦C over Florida during sum- mer conditions incurs an error of about 0.6 µm underestimate in re for re = 14 µm. The error increases slightly for larger re. measurements. This research shows that submicron aerosols, which constitute essentially almost all CCN, can usually be represented as linear mixtures of an organic component with κ ≈0.1 to 0.2 and an inorganic component with κ ≈0.6 (Gun- the et al., 2009; Rose et al., 2011). Where appropriate, an additional sea salt component with a κ ≈1.3 can be intro- duced. This approach has provided a global model of κ dis- tributions that is in very good agreement with observations (Pringle et al., 2010). D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2048 Fig. 7. The CCN spectra can be calculated using measurements or model simulations of Na and S(wb). This chart was drawn using data from P¨oschl et al. (2010) for the activation of organic aerosol particles with κ = 0.13 as observed for pristine tropical rainforest air. κ = 0.3 is more representative for most other continental aerosol regimes. Increasing κ would lower the S values in the ﬁgure (Reut- ter et al., 2009). tributions have larger errors associated with them than κ has, both can be used as priors with speciﬁed weights and uncer- tainties, and an optimal κ and CCN(d) can be derived from the CCN(S) spectrum. 8 Combining Na and Wb for obtaining the CCN spectra Earlier researchers used CCN spectra extensively because they can be obtained directly from in situ measurements with cloud chambers with control on varying the supersaturation. Often, these CCN spectra were parameterized using a power law of the form CCN(S) = CCN (1 %) ×Sb, such that the Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ 10.1 Measurements of thermodynamic parameters that affect clouds Clouds are affected primarily by thermodynamical condi- tions which determine to a large extent where and when they form, their updraft speeds and vertical extent. Aerosols mod- ulate the cloud properties by affecting the cloud drop sizes and ice nucleation process, and in turn the rate of conversion of cloud water to hydrometeor, glaciation, latent heat release and evaporation. This, in turn, affects the cloud dynamics and feeds back to the meteorology. As shown already in Section 6, an error of a factor e in the retrieved re would be ampliﬁed by e3 in the calculated Na. This means that an overestimate error of 1 µm for a cloud with re=15 µm would propagate to a bias error in Na of (16/15)3 = 1.21. When this uncertainty is added to a sim- ilar error of about 20 % due to deviations from the assumed mixing model, the error in Na grows to a factor of 1.45. Retrieving the CCN was described in the previous sec- tions. In addition, the thermodynamic phase (ice versus liq- uid water) of the clouds can be retrieved by the method de- scribed by Martins et al. (2011), using the 2.1 and 2.3 µm wavebands. A number of parameters that are a manifestation of the cloud forcing can be measured by CHASER. Such pa- rameters are: The concentration of CCN can be obtained if the maxi- mum vapor supersaturation at cloud base height, S, is known. This, in turn, can be obtained from cloud base updraft speed, which can be retrieved from very high-resolution (50 m) dual stereoscopic images at oblique view of 30 degrees off nadir for the center camera. Assuming that a cloud protu- berance seen from the three angles can be perfectly matched and tracked over the 2 min (validation of this assumption re- quires further research) and using the beneﬁt of knowledge that cloud base is ﬂat over a well mixed boundary layer can yield vertical motions wb near cloud base with an accuracy of 0.2–0.5 m s−1. According to Eq. (2), a bias error of 20 % in wb causes S to be biased by 12 % in the same direction. When the errors are combined in the same direction, a 45 % bias error in Na causes S to be biased by 14 % in the oppo- site direction. 9.1 Retrieving errors in cloud drop effective radius An alternative approach would be to apply a model that predicts both aerosol size distribution and κ on the basis of on aerosol source/sink and transport processes, such as the Global Model of Aerosol Processes (GLOMAP) (Spracklen et al., 2005a, 2005b, 2010). Although the size dis- www.atmos-meas-tech.net/5/2039/2012/ www.atmos-meas-tech.net/5/2039/2012/ Atmos. Meas. Tech., 5, 2039–2055, 2012 2049 D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations leads to an error of only 10 % in the critical particle diameter of CCN activation (Kreidenweis et al., 2009; P¨oschl et al., 2009), and the actual inﬂuence of κ on cloud droplet num- ber is even smaller because of compensation effects between updraft velocity, aerosol hygroscopicity and water vapor su- persaturation (Reutter et al., 2009). Field measurement data conﬁrm that the prediction of CCN concentration depends much more strongly on the variability of aerosol particle con- centration and size than on the variability of κ (e.g., Gunthe et al., 2009; Rose et al., 2010, 2011). The accuracy of in- dividual CCN retrievals will depend on the accuracy of the available aerosol measurement and modeling data, and will thus vary for different regions and atmospheric conditions. In any case, κ is expected to be one of the least uncertain and least critical aerosol parameters. is negligibly affected from surface properties when its liquid water content exceeds 0.25 g m−3 (Rosenfeld et al., 2004). The required geometrical depth or water content of a cloud with similar re for having the same opacity at 2.1 µm is more than double. Therefore, an effort is made here to retrieve re at both wavelengths and use them in combination. The way by which they will be combined requires additional study. Taking into account the 3-D effects in the 2.1 µm wave- band allows a retrieval accuracy of 1 µm (Zinner et al., 2008). Using the 3.7 µm waveband is likely to increase the accuracy. The quantiﬁcation requires repeating the study of Zinner at al. (2008) in this waveband. 9.2 Error propagation in the calculation of CCN (S, d) Freud et al. (2011) have shown based on aircraft measure- ments of vertical proﬁles of re and Nd that the assumption of extreme inhomogeneous mixing results in systematic over- estimate bias of Na by a factor of about 1.3, when the cloud environment is dry, with RH of about 50 %. Taking into ac- count the actual RH in the vicinity of the cloud may allow decreasing this bias and its uncertainty, as the difference be- tween homogeneous and inhomogeneous mixing vanishes as the environmental RH goes to 100 %. 10.1 Measurements of thermodynamic parameters that affect clouds This allows the retrieval of the CCN(S) with an accuracy of 25 %. 1. The cloud vertical growth rate. This is a manifestation of the atmospheric instability and forcing. It can be ob- tained from the stereoscopic analysis of the MAI. 2. The sensible (Qh) and latent (Qe) surface heat ﬂuxes. The magnitude of the ratio between these ﬂuxes (B = Qh/Qe) determines to a large extent the thermodynamic surface properties and cloud forcing. Over the ocean B is of the order of 0.1, whereas B is 5–10 times larger over land. B can be obtained from comparisons of the surface skin to surface air temperature, where the lat- ter can be calculated using the dry adiabatic lapse rate extended from the cloud base to the surface. This lapse rate can be retrieved because cloud base height, temper- ature and the surface temperature can all be retrieved by CHASER. 2. The sensible (Qh) and latent (Qe) surface heat ﬂuxes. The magnitude of the ratio between these ﬂuxes (B = Qh/Qe) determines to a large extent the thermodynamic surface properties and cloud forcing. Over the ocean B is of the order of 0.1, whereas B is 5–10 times larger over land. B can be obtained from comparisons of the surface skin to surface air temperature, where the lat- ter can be calculated using the dry adiabatic lapse rate extended from the cloud base to the surface. This lapse rate can be retrieved because cloud base height, temper- ature and the surface temperature can all be retrieved by CHASER. In order to obtain CCN(S, d), where d is the aerosol par- ticle diameter, the hygroscopicity growth parameter of the aerosol, κ, must be known. With state-of-the-art global atmo- spheric chemistry and transport models, the average devia- tion between predicted and measured κ values and CCN con- centrations can usually be kept below ∼30 % (e.g., Pringle et al., 2010; Spracklen et al., 2011). An error of 30 % in κ Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ www.atmos-meas-tech.net/5/2039/2012/ This can be ob- tained from tracking layer cloud elements with the MAI, elements which can conﬁdently be assumed not to grow vertically very fast, at various heights. The essence of the difference between the land and ocean surface is the change in the sensible heat ﬂuxes, relative humidity and respective cloud base height, and the resul- tant cloud base updrafts and organization which in turn con- tribute to the difference between the maritime and continen- tal clouds to an unknown extent. Having the measurements of both the aerosols and these thermodynamic properties, along with the relative humidity and cloud vertical motions aloft, will provide a dataset from which it will be possible to dis- entangle the effects of the aerosols from the other effects. The substantiation for this claim is provided in the next sub- section, which addresses the roles of both thermodynamic and aerosols in affecting clouds in ways that are difﬁcult to separate. The plausibility of two possible explanations is under- scored by the ﬁrmly established contrast in cloud conden- sation nuclei concentrations between land and ocean that is found in many regions (Table 3). Oceanic air is often clean, while continental air is polluted in many areas (Hogan, 1977; Schaefer and Day, 1981). This difference (Table 2) is widely attributed to the prevalence of aerosol sources over land (by smoke from ﬁre, air pollution, emissions from vegetation, and lofting of mineral dust and by volcanic eruptions), and the weaker aerosol sources over the sea. The general con- trast in CCN concentrations between polluted and clean re- gions is an order of magnitude (Andreae, 2009). However, the land/ocean contrast in cloud base height is as certain as the contrast between mostly polluted land and clean ocean in CCN (Williams and Satori, 2004; Williams et al., 2005), while the cloud base height is entirely independent of CCN concentrations. The thickness of warm cloud (difference be- tween freezing level height and cloud base height) is nec- essarily generally greater over ocean and favors coalescence and the removal of cloud water as warm rain. The thermody- namic effect of thicker warm cloud over oceans mimics the aerosol effect there of cleaner air, smaller CCN concentra- tion, larger cloud droplets and increased coalescence. Over oceans, the surface sensible heat ﬂux is smaller and so the cloud base updraft strength is smaller (Table 3), nucleating www.atmos-meas-tech.net/5/2039/2012/ 050 D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations Table 2. Contrast in land–ocean behavior in moist convection. Phenomenon Thermodynamic Explanation Aerosol Explanation “Warm rain” convection more prevalent over ocean Reduced time for coalescence in stronger continental updraft Larger droplets over ocean Lightning activity more prevalent over land Greater instability over land and stronger cloud base updraft Enhanced delivery of cloud water to mixed phase region Large hail more prevalent over land Stronger updraft and thinner warm rain region over land Enhanced delivery of cloud water to mixed phase region Upper tropospheric cirrus more prevalent over land More vigorous ice production in stronger updraft over land Greater numbers of small ice particles nucleated over land D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2050 Table 2. Contrast in land–ocean behavior in moist convection. Table 2. Contrast in land–ocean behavior in moist convection. 3. Vertical proﬁle of the temperature and moisture. These properties can be retrieved from the vertical proﬁles of cloud surface temperature and the precipitable water above cloud elements at various heights. 3. Vertical proﬁle of the temperature and moisture. These properties can be retrieved from the vertical proﬁles of cloud surface temperature and the precipitable water above cloud elements at various heights. 3. Vertical proﬁle of the temperature and moisture. These properties can be retrieved from the vertical proﬁles of cloud surface temperature and the precipitable water above cloud elements at various heights. crustal materials and liquid water. The stronger heating of the land surface is responsible for greater air–surface tem- perature contrast and greater instability there (Williams and Renno, 1993), and for the larger updrafts at both cloud base height (Williams and Stanﬁll, 2002) and at higher levels of deep convection (Jorgenson and LeMone, 1989). The rela- tively recent upsurge of interest and observational attention to atmospheric aerosol (Molini´e and Pontikis, 1995; Rosen- feld and Woodley, 2003; Hicks et al., 2005) has led to alter- native physical explanations for the land/ocean contrasts (Ta- ble 2) that depend on the regulation of cloud droplet size and compete with the thermodynamic explanations so strongly that the two effects are difﬁcult to distinguish from each other (Williams et al., 2002; Williams and Stanﬁll, 2002). New satellite-based methods are needed to address both explana- tions. 4. Vertical proﬁle of horizontal winds. www.atmos-meas-tech.net/5/2039/2012/ 11 Summary The scientiﬁc basis for a satellite mission to retrieve CCN concentrations and investigate their impacts on convective clouds is introduced here. It addresses the outstanding prob- lem of cloud-mediated aerosol impacts on cloud radiative and thermodynamic forcing. The approach measures simul- taneously the cloud composition and thermodynamic prop- erties, and infers the number of activated CCN, Na, from the vertical evolution of the retrieved cloud re in convec- tive elements. The Na can be retrieved with an accuracy of at least 45 %. To achieve this goal, multispectral measurements of the sun-illuminated side of the clouds at a resolution of 100 m are planned by the proposed CHASER satellite mis- sion (Renno et al., 2012). The uncertainty in the model of mixing (homogeneous versus extreme inhomogeneous) can add another similar level of uncertainty, but this may be im- proved with assessment of the ambient relative humidity. The disentanglement of thermodynamic and aerosol con- tributions to land/ocean contrasts in moist convection will re- quire the simultaneous measurement of thermodynamic vari- ables (CAPE, sensible heat ﬂux, cloud base height, updraft speed) and the CCN at cloud base height, on the same set of clouds, comparing cloud properties over clean and polluted continental and marine regions. Then sensitivity analyses can be performed based on partial derivatives with respect to key thermodynamic and aerosol variables to quantify the various contributions. Compelling evidence for a role for anthropogenic aerosol in the competition with thermodynamics in modifying moist convection is found in recent ﬁndings on the “weekend ef- fect” (Bell et al., 2009; Rosenfeld and Bell, 2011), which are supported by the review of Sanchez-Lorenzo et al. (2012). In parallel with aerosol measurements showing a deﬁcit con- centration on weekends are ﬁndings that lightning, hail and tornadoes are all suppressed (by 10–20 %) over weekends relative to week days. In the absence of evidence for changes in thermodynamic quantities on the weekly time scale over continents, a distinct role for aerosol remains. The concentration of CCN can be obtained if the maxi- mum vapor supersaturation at cloud base height, S, is known. This, in turn, can be obtained from cloud base updraft speed, which can be retrieved from very high resolution (50 m) dual stereoscopic images at oblique view of 30 degrees off nadir for the center camera. 10.2 The entangled roles of aerosols and meteorology in controlling clouds Comprehensive studies of aerosol effects on light- ning activity on the interannual time scale are lacking. More recent efforts to distinguish thermodynamic from aerosol contributions in explaining local variations in oceanic lightning activity, arriving at the conclusion that aerosol ef- fects are dominant, can be found in Yuan et al. (2011). al., 2009). Comprehensive studies of aerosol effects on light- ning activity on the interannual time scale are lacking. a smaller fraction of the aerosols into cloud droplets and al- lowing more time for coalescence, and again favoring warm rain (Table 2). More recent efforts to distinguish thermodynamic from aerosol contributions in explaining local variations in oceanic lightning activity, arriving at the conclusion that aerosol ef- fects are dominant, can be found in Yuan et al. (2011). Considering now the other three phenomena in Table 2 – lightning, large hail and upper tropospheric cirrus clouds – all three depend on the supply of condensate to the upper cloud. Lightning is mediated by graupel colliding with ice crystals. This graupel forms from supercooled water that has survived the coalescence process. Large hail forms by accre- tion of supercooled water in strong updrafts. Finally, upper tropospheric cirrus cloud depends on the nucleation of ice particles in the mixed phase region that is promoted by strong continental updrafts and by aerosol-rich air. 10.2 The entangled roles of aerosols and meteorology in controlling clouds Contrasts in the behavior of moist convection between land and ocean have been increasingly well quantiﬁed on a global basis by satellite observations over the last twenty years, and are summarized in Table 2. Large hail (Williams et al., 2005), lightning activity (Christian et al., 2003) and upper level cir- rus cloud (Kent et al., 1995) are all substantially more preva- lent over land than over ocean. In contrast, the prevalence of warm rain showers, in which ice particles are absent, is found over oceans (Williams and Stanﬁll, 2002; Liu and Zipser, 2009). The traditional physical explanations for these contrasts, also reported in Table 2, are generally based on thermody- namic differences between land and ocean, which in turn are linked to contrasts in the surface properties (Table 3) of Atmos. Meas. Tech., 5, 2039–2055, 2012 www.atmos-meas-tech.net/5/2039/2012/ D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 205 Table 3. Contrast in land–ocean physical and meteorological properties. Property Land Ocean Surface heat capacity Small (0.2–0.5 cal g m−1 ◦C−1), immobile surface Large (1 cal g m−1 ◦C−1), mobile surface Air-surface temp. contrast large (1–10 ◦C) Small (∼1 ◦C) Bowen ratio Large (0.2–1) Small (0.1) Boundary layer CCN High in polluted regions (>1000 cm−3) Low in clean regions (∼100 cm−3) Cloud base height High (1000–4000 m) Low (500 m) Thickness of “warm” cloud Small (0–3000 m) Large (∼4000 m) Cloud base updraft speed High (∼5 m s−1) Small (∼2 m s−1) CAPE Larger (0–3000 j kg−1) Smaller (0–2000 j kg−1) Convective updraft speed High (>10 m s−1) Low (<10 m s−1) D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 205 Table 3. Contrast in land–ocean physical and meteorological properties. Property Land Ocean Surface heat capacity Small (0.2–0.5 cal g m−1 ◦C−1), immobile surface Large (1 cal g m−1 ◦C−1), mobile surface Air-surface temp. contrast large (1–10 ◦C) Small (∼1 ◦C) Bowen ratio Large (0.2–1) Small (0.1) Boundary layer CCN High in polluted regions (>1000 cm−3) Low in clean regions (∼100 cm−3) Cloud base height High (1000–4000 m) Low (500 m) Thickness of “warm” cloud Small (0–3000 m) Large (∼4000 m) Cloud base updraft speed High (∼5 m s−1) Small (∼2 m s−1) CAPE Larger (0–3000 j kg−1) Smaller (0–2000 j kg−1) Convective updraft speed High (>10 m s−1) Low (<10 m s−1) D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2051 al., 2009). D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations J. Roy. Meteorol. Soc., 131, 1171–1190, doi:10.1256/qj.03.180, 2005. Brenguier, J. L., Pawlowska, H., Schuller, L., Preusker, R., Fis- cher, J., and Fouquart, Y.: Radiative properties of boundary layer clouds: Droplet effective radius versus number concentration, J. Atmos. Sci., 57, 803–821, 2000. Burnet, F. and Brenguier, J. L.: Observational study of the entrainment-mixing process in warm convective clouds, J. At- mos. Sci., 64, 1995–2011, 2007. In summary, the ability to retrieve simultaneously the cloud microphysical and thermodynamic properties and the aerosols in the underlying boundary layer will enable a ma- jor improvement in the accuracy of aerosol indirect radiative forcing and reduce substantially the magnitude of our uncer- tainty, which presently dominates the overall uncertainty in the magnitude of anthropogenic climate forcing. Chen, T. C., Xue, L., Lebo, Z. J., Wang, H., Rasmussen, R. M., and Seinfeld, J. H.: A comprehensive numerical study of aerosol- cloud-precipitation interactions in marine stratocumulus. Atmos. Chem. Phys., 11, 9749–9769, doi:10.5194/acp-11-9749-2011, 2011. Christian, H. J., Blakeslee, R. J., Boccippio, D. J., Boeck, W. L., Buechler, D. E., Driscoll, K. T., Goodman, S. J., Hall, J. M. Koshak, W. J., Mach, D. M., and Stewart, M. F.: Global fre- quency and distribution of lightning as observed from space by the Optical Transient Detector, J. Geophys. Res., 108, 4005, doi:10.1029/2002JD002347, 2003. Acknowledgements. The idea of retrieving CCN from space was stimulated by the discussions at the working group of remote sens- ing cloud–aerosol–precipitation–climate interactions, sponsored by the International Space Science Institute in Bern, Switzerland. We thank NASA Glenn Research Center, the University of Michigan, and DLR for supporting the development of the CHASER Mission concept. Many engineers, managers and technical staff played important roles on the development of CHASER. We also thank Vanderlei Martins, whose cloud side scanner inspired the develop- ment of the CHASER mission. Fan J., Yuan, T., Comstock, J. M., Ghan, S., Khain, A., Le- ung, R. L., Li, Z., Martins, V. J., and Ovchinnikov, M.: Dom- inant role by vertical wind shear in regulating aerosol effects on deep convective clouds, J. Geophys. Res., 114, D22206, doi:10.1029/2009JD012352, 2009. Fan, J., Rosenfeld, D., Ding, Y., Leung, L. R., and Li, Z.: Potential aerosol indirect effects on atmospheric circulation and radiative forcing through deep convection. Geophys. Res. Lett., in press, doi:10.1029/2012GL051851, 2012. Edited by: A. Macke Feingold, G., Yang, S., Hardesty, R. M., and Cotton, W. D. Rosenfeld etal.: Satellite mission to retrieve CCN concentrations 2052 sources and aerosol transport models with an accuracy of bet- ter than 30 %. For a given CCN(S), this allows the calcula- tion of the size spectra of the CCN aerosols to an accuracy of better than 10 %, CCN(d) is an essential aerosol property for simulating aerosol effects on cloud microstructure and precipitation forming processes. The estimates of accuracy should be reﬁned based on simulations and validation against actual aircraft campaigns that simulate the satellite measure- ments and provide in situ validation of the cloud and CCN properties. Andreae, M. O., Rosenfeld, D., Artaxo P., Costa, A. A., Frank, G. P., Longo, K. M., and Silva-Dias, M. A. F.: Smoking rain clouds over the Amazon, Science, 303, 1337–1342, 2004. Andreae, M. O., Jones, C. D., and Cox, P. M.: Strong present-day aerosol cooling implies a hot future, Nature, 435, 1187–1190, 2005. Andreae, M. O.: Correlation between cloud condensation nu- clei concentration and aerosol optical thickness in remote and polluted regions, Atmos. Chem. Phys., 9, 543–556, doi:10.5194/acp-9-543-2009, 2009. Bell, T. L., Rosenfeld, D., Kim, K.-M., Yoo, J. M., Lee, M. I., and Hahnenberger, M.: Midweek increase in U.S. summer rain and storm heights suggests air pollution invigorates rainstorms, J. Geophys. Res., 113, D02209, doi:10.1029/2007JD008623, 2008. The aerosol properties are retrieved from the cloudy boundary layer, where other satellite-based methods of di- rect aerosol measurements are blind. This method is also free of problems that have plagued direct aerosol measurements, such as swelling of the particles at high RH, cloud contami- nation, and aerosols that are not at the same level as the cloud base. Bell, T. L, Rosenfeld, D., and Kim, K.-M.: Weekly cycle in light- ning: Evidence of storm invigoration by pollution, Geophys. Res. Lett., 36, L23805, doi:10.1029/2009GL040915, 2009. Bennartz, R.: Global assessment of marine boundary layer cloud droplet umber concentration from satellite, J. Geophys. Res., 112, D02201, doi:10.1029/2006JD007547, 2007. Retrieving the vertical microstructure and thermodynamic properties of the clouds as well as the horizontal and vertical cloud motions allows obtaining simultaneous CCN concen- trations, dynamics and thermodynamics of the clouds. This will make it possible to address effectively the outstanding challenge of disentangling meteorology from the aerosol ef- fects on clouds, and hence the aerosol radiative and thermo- dynamic climate forcing. Section 9 introduces an approach for doing that. Blyth, A. M., Lasher-Trapp, S. G., and Cooper, W. A.: A study of thermals in cumulus clouds, Q. 11 Summary The analysis with the beneﬁt of knowl- edge that cloud base is ﬂat over a well mixed boundary layer can yield vertical motions, wb, near cloud base with an accu- racy of 0.2–0.5 m s−1. When the retrieval errors of Na are combined in the same direction, a 45 % bias error in Na causes S to be biased by 14 % in the opposite direction. This allows the retrieval of the CCN(S) with an accuracy of 25 %. 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https://openalex.org/W3207750568_1	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	1,357	2,355	Alcántara, M., García, M. y Rivas, C. (2020). Politics and Political Elites in Latin America. Challenges and Trends. Cham: Springer. 353 pp. Luis Antonio González1 DOI: https://doi.org/10.15446/cp.v16n31.97928 Por más de 25 años, el Proyecto de Élites Parlamentarias en América Latina de la Universidad de Salamanca (PELA-USAL), dirigido por el profesor Manuel Alcántara, ha llenado un vacío en los estudios de opinión enfocados en las élites políticas. El proyecto busca conocer quiénes son los legisladores, cuál ha sido su trayectoria y qué opiniones, valores y creencias tienen con respecto a temas diversos como la confianza institucional, la satisfacción con la democracia, las relaciones ejecutivo-legislativo, la política exterior, el género o la ideología personal y partidista. La relevancia de estudiar las élites políticas deriva no solo del papel central que desempeñan estos actores dentro de sus funciones formales (deliberación de la agenda pública, canalización de demandas ciudadanas, elaboración de políticas, revisión y aprobación del presupuesto, control político, etc.), sino de su influencia, cada vez mayor desde la década de los noventa en la toma de decisiones. Con información recabada en el PELA-USAL se han publicado numerosos artículos científicos, capítulos y libros. Su libro más reciente es Politics and Political Elites in Latin America. Challenges and Trends, editado por Manuel Alcántara, Mercedes García Montero y Cristina Rivas. Esta obra recoge todos los años de experiencia del proyecto para analizar temas de relevancia política en América Latina desde la perspectiva de los legisladores nacionales, por lo que constituye el esfuerzo más importante y enriquecedor de los últimos años en el estudio de las élites políticas en la región. 1 Profesor asistente del departamento de Ciencia Política y Relaciones Internacionales, Universidad del Norte, Barranquilla, Colombia. Correo: ltule@uninorte.edu.co. ORCID: https://orcid.org/0000-0002-3336-8640 Este artículo está publicado en acceso abierto bajo los términos de la licencia Creative Commons Reconocimiento-NoComercial-SinObraDerivada 2.5 Colombia. \| VOL. 16, N.º 31 ENE-JUN 2021 · ISSN IMPRESO 1909-230X · EN LÍNEA 2389-7481 /PP. 301-304 301 LUIS ANTONIO GONZÁLEZ El capítulo introductorio, escrito por Mélany Barragán, Cristina Rivas y José Manuel Rivas, presenta las bases teórico-metodológicas que guían el libro. Los autores retoman los principales debates en el estudio de la élite política, para después adentrarse en los aspectos metodológicos y la evolución del proyecto, así como en los retos que implica la definición del diseño del cuestionario y de la unidad de análisis, la realización del trabajo de campo y la exploración de datos. A partir del siguiente capítulo el libro se divide en dos partes. La primera se compone de estudios que abordan cuestiones sobresalientes de la política latinoamericana como la calidad de la democracia, la carrera de los legisladores, las relaciones ejecutivo-legislativo y la participación y representación de las mujeres. Abriendo esta sección, Theresa Kernecker aborda un tema central en los estudios legislativos como la carrera política de los congresistas. Con base en las preferencias de los legisladores de 17 países entre 2006 y 2012, Kernecker contrasta el argumento predominante en la literatura latinoamericana sobre el deseo de los representantes de continuar con su carrera fuera del recinto legislativo (progressive ambition). En el tercer capítulo, Cristina Rivas y Manuel Alcántara analizan la confianza de los legisladores latinoamericanos en las principales instituciones políticas. A través del índice de confianza política los autores observan si la confianza en el Congreso, presidente, poder judicial y partidos políticos se mantiene estable o ha ido cambiando conforme al desempeño político y económico de los gobiernos. En una línea similar, Ignacio Arana y Carolina Guerrero, en el Capítulo 7, indagan en torno a los factores que explican la confianza de los legisladores en el presidente en turno. Por su parte, Mar Martínez Rosón y Araceli Mateos comparan información del PELA-USAL con el barómetro de las Américas de la Universidad de Vanderbilt. Este trabajo es imprescindible para identificar similitudes y diferencias entre la élite política y la ciudadanía con relación a la satisfacción en la democracia. Otro capítulo que combina datos de dos proyectos es el de Asbel Bohigues y Manuel Alcántara. Aquí los autores utilizan bases de datos del PELA-USAL y V-Dem para identificar posibles relaciones entre las opiniones de las élites políticas y las variedades de las democracias. El Capítulo 5, escrito por Margarita Corral y Mar Martínez Rosón, está dedicado al estudio de las instituciones informales y su relación con variables socioeconómicas, la carrera política y el acceso a recursos por 302 \| ISSN IMPRESO 1909-230X • EN LÍNEA 2389-7481 • UNIVERSIDAD NACIONAL DE COLOMBIA • FACULTAD DE DERECHO, CIENCIAS POLÍTICAS Y SOCIALES • DEPARTAMENTO DE CIENCIA POLÍTICA RECENSIONES parte de los legisladores. Uno de los principales hallazgos de estas autoras es que los legisladores con experiencia política previa son más proclives a ofrecer beneficios particulares y la relación es más significativa cuando forman parte del partido gobernante. Más adelante, el trabajo de Mercedes García Montero y Cecilia Rodríguez se centra en el género y la participación política de las legisladoras latinoamericanas. En este, las investigadoras del Instituto de Iberoamérica exploran si algunas variables institucionales vinculadas al género – como la experiencia, carrera o dedicación política– explican los perfiles de las legisladoras. Desde otra perspectiva, Asbel Bohigues y Scott Morgenstern analizan la percepción de los congresistas sobre Estados Unidos y China a través de dos dimensiones: la influencia de estos países en la región y la confianza percibida hacia sus gobiernos. A los autores les interesa saber si la opinión de las élites latinoamericanas sobre ambas potencias está relacionada y hasta qué punto el rechazo o apoyo pueden ser complementarios. La segunda parte del libro profundiza en estudios de caso y destaca las actitudes de los legisladores con respecto a la ideología, la democracia y las instituciones políticas. Carlos Ranulfo Melo, Manoel Leonardo Santos y Rafael Câmara comparan tres países estructuralmente diferentes, pero con trayectoria dictatorial militar similar, como Brasil, Chile y Uruguay, con la finalidad de identificar hasta qué punto la autoubicación ideológica de los diputados está relacionada con la percepción que se tiene respecto de otros temas sustantivos. En el siguiente capítulo, Cristian Márquez y Patricia Marenghi analizan la variabilidad del apoyo a la democracia por parte de los legisladores mexicanos. Este tipo de estudios permite conocer la satisfacción con la democracia, establecer niveles de congruencia entre las preferencias de la élite y la ciudadanía, e identificar patrones de cambio o estabilidad en estas a lo largo de más de dos décadas. Adriana Ramírez Baracaldo y José Manuel Rivas Otero, por su parte, se enfocan en la opinión de la oposición parlamentaria colombiana con la finalidad de indagar en los factores que influyen para que los legisladores se identifiquen como parte de ésta. Según Ramírez y Rivas, dentro de los factores más significativos para sentirse parte de la oposición está la afiliación partidista, la ideología y el nivel de confianza en el presidente. Daniel Chasquetti y Lucía Selios observan, para el caso uruguayo, que las opiniones de los legisladores en este país se han mantenido 16 (31) 2021 ENERO-JUNIO • PP. 301-304 303 LUIS ANTONIO GONZÁLEZ relativamente estables durante cerca de veinte años. Esta regularidad tiene que ver con el hecho de que Uruguay mantiene un sistema de partidos institucionalizado y una estructura de competencia favorable a la estabilidad en las opiniones de los legisladores. Esto último es resultado de una clara identificación ideológica del elector con los partidos políticos. El libro cierra con el capítulo de Mélany Barragán, donde se comparan las actitudes de las élites parlamentarias en Guatemala y Panamá para evidenciar diferencias sustantivas en aspectos variados del sistema político entre estos dos países centroamericanos. En conjunto, el libro deja entrever que las actitudes y los valores de la élite política son importantes. Estos constituyen un sustento empírico de gran valor para estudiar temas tan variados de las democracias representativas como la profesionalización de la política, el género, las relaciones ejecutivo-legislativo, la ideología o la confianza institucional. La riqueza temática de la obra se complementa con las diferentes aproximaciones metodológicas y perspectivas teóricas que los autores imprimen a cada uno de sus capítulos. 304 \| ISSN IMPRESO 1909-230X • EN LÍNEA 2389-7481 • UNIVERSIDAD NACIONAL DE COLOMBIA • FACULTAD DE DERECHO, CIENCIAS POLÍTICAS Y SOCIALES • DEPARTAMENTO DE CIENCIA POLÍTICA.	19,321
https://openalex.org/W3185847377	OpenAlex	Open Science	CC-By	2,021	Determining soil particle-size distribution from infrared spectra using machine learning predictions: Methodology and modeling	Élizabeth Parent	English	Spoken	8,688	17,322	RESEARCH ARTICLE Determining soil particle-size distribution from infrared spectra using machine learning predictions: Methodology and modeling Elizabeth Jeanne ParentID, Serge-E´ tienne ParentID, Le´on Etienne Parent Department of Soils and Agrifood Engineering, Universite´ Laval, Que´bec, Canada elizabeth.parent.1@ulaval.ca a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 Editor: Debjani Sihi, Emory University, UNITED STATES Received: April 29, 2020 Accepted: May 16, 2021 Published: July 20, 2021 Received: April 29, 2020 Accepted: May 16, 2021 Published: July 20, 2021 Copyright: © 2021 Parent et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: Data and R code elaborated by Elizabeth Parent and Serge-Etienne Parent are available at: https://github.com/ eliparent/particle-size-distribution-Ft-NIR- spectroscopy. Abstract Accuracy of infrared (IR) models to measure soil particle-size distribution (PSD) depends on soil preparation, methodology (sedimentation, laser), settling times and relevant soil fea- tures. Compositional soil data may require log ratio (ilr) transformation to avoid numerical biases. Machine learning can relate numerous independent variables that may impact on NIR spectra to assess particle-size distribution. Our objective was to reach high IRS predic- tion accuracy across a large range of PSD methods and soil properties. A total of 1298 soil samples from eastern Canada were IR-scanned. Spectra were processed by Stochastic Gradient Boosting (SGB) to predict sand, silt, clay and carbon. Slope and intercept of the log-log relationships between settling time and suspension density function (SDF) (R2 = 0.84–0.92) performed similarly to NIR spectra using either ilr-transformed (R2 = 0.81–0.93) or raw percentages (R2 = 0.76–0.94). Settling times of 0.67-min and 2-h were the most accurate for NIR predictions (R2 = 0.49–0.79). The NIR prediction of sand sieving method (R2 = 0.66) was more accurate than sedimentation method(R2 = 0.53). The NIR 2X gain was less accurate (R2 = 0.69–0.92) than 4X (R2 = 0.87–0.95). The MIR (R2 = 0.45–0.80) performed better than NIR (R2 = 0.40–0.71) spectra. Adding soil carbon, reconstituted bulk density, pH, red-green-blue color, oxalate and Mehlich3 extracts returned R2 value of 0.86– 0.91 for texture prediction. In addition to slope and intercept of the SDF, 4X gain, method and pre-treatment classes, soil carbon and color appeared to be promising features for rou- tine SGB-processed NIR particle-size analysis. Machine learning methods support cost- effective soil texture NIR analysis. RESEARCH ARTICLE Determining soil particle-size distribution from infrared spectra using machine learning predictions: Methodology and modeling Elizabeth Jeanne ParentID*, Serge-E´ tienne ParentID, Le´on Etienne Parent Department of Soils and Agrifood Engineering, Universite´ Laval, Que´bec, Canada RESEARCH ARTICLE OPEN ACCESS Citation: Parent EJ, Parent S-t, Parent LE (2021) Determining soil particle-size distribution from infrared spectra using machine learning predictions: Methodology and modeling. PLoS ONE 16(7): e0233242. https://doi.org/10.1371/ journal.pone.0233242 Editor: Debjani Sihi, Emory University, UNITED STATES PLOS ONE PLOS ONE PLOS ONE PLOS ONE Particle-size distribution predicted by infrared spectroscopy otherwise, the drag force for sedimentation increases faster than predicted by Stokes’ law [3]. Because organic matter binds soil particles [4], organic matter is often destroyed using a perox- ide pre-treatment as method modifier to disperse soil particles. Laser techniques are much faster than sedimentation techniques but tend to underestimate clay-size particles due to insuf- ficient particle dispersion despite sonication pre-treatment, and this may require using conver- sion equations [5,6]. Inc., Groupe Gosselin FG, Prochamps Inc., and Ferme Daniel Bolduc Inc. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This does not alter our adherence to PLOS ONE policies on sharing data and materials. On the other hand, visible and near infrared (VIS–NIR: 350–2500 nm) spectroscopy is an efficient soil quality and fertility screening tool [7] because spectra correlate well with several chemical, physical and mineralogical properties [8]. The VIS represents the visible light range between 350 and 780 nm, from violet to red. Because most routine soil and plant laboratories are equipped with NIR spectrometers for forage analysis, they may contribute to documenting soil characteristics at low cost. Where VIS is not available, soil RGB (red-green-blue) can be assessed from the Munsell color chart using computer models (“munsell2rgb” from the “aqp” package). Mid-infrared (MIR) is generally more accurate than NIR [9] but requires more sam- ple preparation, limiting its application as routine determination method. Infrared spectroscopy (IRS) more accurately predicts clay than sand and silt contents [10] because the IR spectrum is sensitive to clay mineralogy [10–12] and total reflectance decreases as grain size increases [13,14]. Light absorption is also influenced by soil features such as parti- cle roundness [15–18], soil pH, and the Mehlich-3 soil test for Ca, Mg and Mn [19]. The IRS detects Al-OH (2200 nm) and Fe-OH (2290 nm) [20,21] that in turn have an impact on soil structure [22] and IR reflectance [23]. The VIS-NIR spectra are sensitive to soil moisture and C content [24]. Organic matter, multi-nutrient extraction and reconstituted bulk density from scooped soil samples are also common features quantified in routine laboratories. Competing interests: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This does not alter our adherence to PLOS ONE policies on sharing data and materials. Introduction Soil particle-size distribution (PSD) is of prime importance for plant growth and soil manage- ment [1]. Mimicking particle sedimentation in natural water bodies, PSD has been tradition- ally quantified using the sieve-pipette method that determines particle mass, and the sieve- hydrometer or sieve-plummet balance method that measures changes in suspension density [2]. Sedimentation techniques are thought to overestimate the concentration of plate-like clay particles that do not fit into Stokes’ law [3]. Reynolds’ number should be less than 0.05, Funding: LEP NSERC-CG-2254 Le´on Etienne Parent http://www.vrrc.ulaval.ca/fileadmin/ulaval_ ca/Images/recherche/bd/chercheur/fiche/51840. html This project was funded by the Natural Sciences and Engineering Council of Canada (CG- 2254 and CRDPJ 385199-09), Cultures Dolbec 1 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 PLOS ONE The sedimentation methods provide percentages of sand, silt, and clay from the log-log relationship between settling time and suspension density. The slope and intercept return pro- portions of sand, silt and clay at pre-selected settling times that may vary between laboratories [1], thus affecting the accuracy of IRS models. Providing more flexibility using the slope and intercept of the log-log relationship and reducing the arbitrariness of the settling time selection may allow increased reliability of IRS models calibrated against sedimentation methods. There are unattended sources of error in IRS calibration. There is systematic negative covariance between sand, silt and clay fractions due to resonance within the ternary diagram [25]. Indeed, there are D-1 degrees of freedom in a D-part composition [26]. Not considering the problem of closure to 100% in statistical analysis, confidence intervals about means of pro- portions may take values outside of the compositional space, i.e. < 0 or > 100% [27], and the measures of distance and dissimilarity are non-Euclidian [28]. To return unbiased statistical results, orthonormal balances among subsets of components can be computed as D-1 isomet- ric log ratios (ilr) [29]. The back-transformed ilr values allow recovering proportions of sand, silt, and clay totalling exactly 100% within the limits of the ternary diagram. For both the sedimentation and laser techniques there are several soil pre-treatments (per- oxide, sodium hypochlorite, sodium hexametaphosphate, sonication intensity), soil features, calibration techniques, options (NIR 2X, NIR 4X, settling times or suspension density function for sedimentation; pump and stirrer spin, refractive index of the medium, real or imaginary refractive index, density for laser) and expressions (percentages, ratios) that influence results of particle-size distribution. Machine learning (ML) is an emerging data mining technique of artificial intelligence that can unravel patterns and rules in large data sets [30] and predict target variable from input data [31]. Machine learning methods can account for numerous independent variables that may impact on NIR spectra to assess accurately soil particle-size distribution. 2 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 PLOS ONE Particle-size distribution predicted by infrared spectroscopy This paper is presented in two parts, one focusing on methodology and the other on model- ing. In the methodology section, we hypothesized that methods and pre-treatments return dif- ferent results and thus cannot be combined to run a single IRS calibration set without input information on methodologies. In the modeling section, we hypothesized that: 1. PLOS ONE IRS cannot accurately predict PSD from laser methods due to underestimation of the clay fraction; 2. IRS accurately predicts sand fraction determined using sand sieving methods; 3. IRS more accu- rately predicts PSD from sedimentation methods after accounting for soil features; 4. IRS pre- diction accuracy for PSD is improved using the slope and intercept of the log-log relationship and isometric log ratios compared to raw percentages and pre-determined settling times; and 5. MIR prediction accuracy for PSD is higher compared to the NIR spectra. The objective of this study was to calibrate infrared determination methods against routine PSD methods, methodological modifiers and soil features to reach high IR-ML model accuracy for routine soil texture determination. Characterization of soils The data set of 1298 soil samples collected in the arable layer (0–20 cm) was obtained from sev- eral research institutions in Que´bec, Canada (Fig 1). The main crops were maize (Zea mays) cereals and forages on coarse- to fine-textured soils, potatoes (Solanum tuberosum) on sandy loams and loamy sands, and cranberries (Vaccinium macrocarpon) on sandy soils. The soils were mainly Inceptisols and Spodosols. Soil samples were air-dried then passed through a 2-mm sieve [32,33]. The 2-mm sieved soil was 3-mL scooped and then weighed to determine the reconstituted bulk density as per- formed routinely in soil testing laboratories. Soil color was assessed on dry samples using the Munsell chart and then transformed into RGB percentages using “munsell2rgb” in R. The Fig 1. Repartition of soil sampling areas on eastern provinces in Canada. Map tiles by Stamen Design, under CC BY 3.0. Data by OpenStreetMap, under ODbL. Contains information from OpenStreetMap and OpenStreetMap Foundation, which is made available under the Open Database License. http://maps.stamen.com/#toner/12/37.7706/-122.3782. https://doi org/10 1371/journal pone 0233242 g001 Fig 1. Repartition of soil sampling areas on eastern provinces in Canada. Map tiles by Stamen Design, under CC BY 3.0. Data by OpenStreetMap, under ODbL. Contains information from OpenStreetMap and OpenStreetMap Foundation, which is made available under the Open Database License. http://maps.stamen.com/#toner/12/37.7706/-122.3782. htt //d i /10 1371/j l 0233242 001 https://doi.org/10.1371/journal.pone.0233242.g001 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 3 / 16 PLOS ONE Particle-size distribution predicted by infrared spectroscopy total carbon (Ct) was quantified using the Leco CNS analyzer (Leco Corporation, St. Joseph, Michigan). For pH determination, 10 g of soil was mixed with 20 mL 0.01M CaCl2. For oxalate extracts, a 0.5 g sample of soil was mixed with 20 mL of oxalate solution (0.2 M of ammonium oxalate and 0.2 M of oxalate acid) and agitated for 4 h in the dark [34]. The mixture was centri- fuged at 2000 rpm for 5 min and then filtered through Whatman no. 40 paper. Concentrations of P, Fe, Ca, Al, Mn and Si were quantified by ICP-OES. Soils were also extracted using the routine Mehlich3 method [35]. Concentrations of P, Ca, Mg, Fe, Al, Mn, Zn and Cu were quantified by ICP-OES. Particle-size analysis Particle size distribution was analyzed in 50–100 g samples using the sedimentation method [36]. A separate batch of soils was pre-treated with peroxide for comparison with the no-per- oxide pre-treatment. Samples were mixed with 0.05 M hexametaphosphate and agitated at 300 rpm for 16 h. The mixture was transferred to a 1 L cylinder and hand-shaken for 30 sec. Sus- pension density readings (g L-1) taken after 0.75-, 5-, 120-, 420-, and 1440- min were referred to as the sedimentation multi-point method. Samples reporting suspension density after 0.67 and 120 min as originally suggested [37] were assigned to the 2-point 2-h sedimentation method. The clay fraction was also recorded after 7-h settling time, close to the 6-h settling time used by Gee and Bauder (1979) [38]. Although the clay fraction was overestimated as compared to the 7-h sedimentation multi-point method, the 2-point 2-h sedimentation method was selected as our reference because it has been widely used as a proximate method in soil surveys combined with tactile assessment. After taking the last reading, the entire hydrometer contents from sedimentation method were passed through 1-, 0.5-, 0.25-, 0.10- and 0.05- mm sieves under tap water to clean the coarser particles of any adhering finer parti- cles and to determine the sand-size distribution. The suspension density was transformed into particle-size percentages by mass using standard equations [37,38]. The sedimentation curve relating suspension density to settling time was log-transformed (ln) to determine the slope and intercept as model parameters. Samples were also analyzed using the Mastersizer 2000 Laser particle size analyzer (Malvern Instruments, Worcestershire, UK, measurements at 633 nm and 466 nm) combined with Hydro 2000G (800-mL tap water volume, 500-rpm stirrer and 2000-rpm pump) with or without ultrasonic action at nominal 40 kHz frequency for 2 min [39]. Only samples within 10 to 20% of obscuration were retained in the database. The refractive index of the medium was set at 1.5. PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 Spectral data acquisition The air-dried and 2-mm sieved samples [40] were placed into a 5-cm quartz cup then scanned using a Nicolet Antaris FT-NIR analyzer (Thermo Electron Corp., Ann Arbor, Michigan). Absorbance was measured with gains of 2X or 4X. Triplicated spectra were scanned 30 times in the range of 9090 to 4000 cm-1 (1100 to 2500 nm) at a resolution of 2 cm-1 (0.3 nm at 1250 nm) [41]. A library of 3069 NIR spectra with 2X gain and 668 spectra with 4X gain were obtained after probabilistic quotient normalization and first derivation [42]. All samples were stored in white plastic storage containers. The soil and KBr samples were ground to less than 74 μm using a mortar and pestle [43], then oven-dried at 105˚C for 3 h [44]. A 2 g sample was weighed and mixed with 200 g KBr powder. The mixture was pressed at 517 MPa for 3 min using a manual hydraulic press (Carver, Model 4350.L, Carver Inc., Wabash, Indiana) to yield clear 13-mm-diameter pellets. The mid-infrared spectra were scanned using a transmission DTGS detector Varian 1000 FT-IR Scimitar series spectrometer (Varian Inc., Palo Alto, California). Spectra were in the range of 4000 to 400 cm-1 (2500 to 25 000 nm) at 4 cm-1 (0.6 nm) resolution. Each pellet was scanned 10 times and rotated once manually at 90˚. A library of 413 MIR spectra was obtained. Calculation The three particle-size fractions and carbon content were isometric log ratio (ilr) transformed as follows [29]: ilr carbonjclay:silt:sand ½ ¼ ffiffiffi 2 3 r ln ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi clay silt sand p carbon ! ð1Þ ilr clayjsilt:sand ½ ¼ ffiffiffi 2 3 r ln ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi silt sand p clay ! ð2Þ ilr siltjsand ½ ¼ ffiffiffi 1 2 r ln silt sand ð3Þ ilr carbonjclay:silt:sand ½ ¼ ffiffiffi 2 3 r ln ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi clay silt sand p carbon ! ð1Þ ð1Þ ilr clayjsilt:sand ½ ¼ ffiffiffi 2 3 r ln ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi silt sand p clay ! ð2Þ ilr siltjsand ½ ¼ ffiffiffi 1 2 r ln silt sand ð3Þ ilr clayjsilt:sand ½ ¼ ffiffiffi 2 3 r ln ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi silt sand p clay ! ð2Þ ð2Þ ilr siltjsand ½ ¼ ffiffiffi 1 2 r ln silt sand ð3Þ ð3Þ PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 4 / 16 4 / 16 PLOS ONE Particle-size distribution predicted by infrared spectroscopy Soil characterization There was a large spectrum of soil properties (Table 1). In comparison, soils of the region have been reported to range from 5 to 137 g organic matter kg-1, 560 to 19 992 mg Feoxalate kg-1 and 270 to 36 638 mg Aloxalate kg-1, and 4–494 g sand kg-1, 40–734 g silt kg-1, and 6–796 g clay kg-1 [47]. Particle-size distribution is presented in Fig 2. Orthonormal balances were normally dis- tributed except for [clay \| silt,sand] using the laser method (Fig 3). Indeed, the clay content was systematically underestimated, and the silt content systematically overestimated by the laser method. Spectral data modeling Statistical analyses were conducted using the R-3.6.1 version [45] with the packages “tidy- verse”, “reshape2”, “stringi”, “signal”, “mvoutlier”, “caret”, “compositions”, “soiltexture”, “aqp”, “patchwork”, “broom”, “ggExtra”, “ggmap”, “neuralnet” and “outliers”. The machine learning (ML) method was GBM (Stochastic Gradient Boosting) that outperformed other ML models including neural networks. Neural networks was tested with a hidden of 3 and an acti- vation function settled at “logistic” (S2 Table). The NIR and MIR spectra were normalized and passed through a binning process with 10 cut points and a Savitsky-Golay smoothing filter. After first derivation, spectra compressed into scores using principle component analysis (PCA) [46]. Because of the diversity of the soil samples, we did not remove outliers. Sand, silt and clay percentages, oxalate and Mehlich-3 extracts composed of 0.2N CH3COOH-0.25N NH4N03-0.015NNH4F-0.013NHN03-0.001M EDTA were ilr-transformed. Every set was divided into training (70%) and testing (30%). Methodologies Paired t-tests comparing methods are presented in Table 2. Comparisons involving the perox- ide pre-treatment must be interpreted with care due to the smaller number of observations (26 to 38). Mean ilr differences between the reference 2-point 2-h sedimentation method and other methods are illustrated in Fig 4. The difference between the laser and the 2-point 2-h 5 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 Particle-size distribution predicted by infrared spectroscopy PLOS ONE Particle size distribution of studied soils in the Canadian textural diagram. https://doi.org/10.1371/journal.pone.0233242.g002 sand sieving. Sand contents did not differ significantly between the peroxide pre-treatments or multi-points calculation without pre-treatment. Except for the peroxided pre-treated dataset, the [carbon \| clay,silt,sand] balances were differentially influenced by methodologies. sand sieving. Sand contents did not differ significantly between the peroxide pre-treatments or multi-points calculation without pre-treatment. Except for the peroxided pre-treated dataset, the [carbon \| clay,silt,sand] balances were differentially influenced by methodologies. PLOS ONE Table 1. Ranges of soil properties (0–20 cm) in the data set (particle-size distribution according to the multi-point 7-h sedimentation method using 45-sec settling time for sand and 7-h settling time for clay). Site Mean Standard deviation Minimum Maximum pH (0.01 M CaCl2) 5.36 0.67 3.37 7.90 g kg-1 Sand total 613 24 1 986 • 1–2 mm 41 50 0 282 • 0.5–1.0 mm 108 115 1 541 • 0.25–0.5 mm 191 126 5 580 • 0.1–0.25 mm 235 154 3 766 • 0.25–0.01 mm 118 103 0 587 Silt 256 15 5 816 Clay 131 13 5 839 Total C 25.5 2.4 0.1 443 Total N 1.9 1.6 0.7 19 Total S 0.3 0.4 0 10.3 mg kg-1 Feoxalate 5599 2449 447 10400 Aloxalate 4000 2693 528 15346 Mnoxalate 309 304 7 2287 Caoxalate 30 14 3 153 Poxalate 803 464 68 2266 Sioxalate 829 578 88 5585 PMehlich3 118 62 6 362 CaMehlich3 1776 1437 113 5103 MgMehlich3 191 224 5 1572 FeMehlich3 232 108 72 715 AlMehlich3 1265 431 310 2241 MnMehlich3 25 20 5 206 ZnMehlich3 8 9 0 130 CuMehlich3 4 3 0.4 30 % Green 47 8 28 68 Blue 34 10 11 58 Red 54 7 31 75 g cm-3 Reconstituted bulk density 1.06 0.16 0.71 1.46 https://doi.org/10.1371/journal.pone.0233242.t001 sedimentation methods depended on the ilr coordinate (also shown in Fig 3). After 2-min of sonication, the [silt \| sand] balance differed slightly between methods while the [clay \| silt, sand] balance differed markedly between methods, indicating insufficient dispersion of clay- size particles. As expected, the multi-point 7-h sedimentation method returned a smaller proportion of clay-size particles compared to the 2-point 2-h sedimentation method due to longer settling time. The peroxide pre-treatment for the multi-point 7-h sedimentation method tended to increase clay and silt fractions. The [silt \| sand] balance resulting from the no-peroxide pre- treatment preceding sedimentation methods was comparable across methods, with the excep- tions of the laser and pretreated multi-point 7-h sedimentation methods. Excluding the 0-min sonicated laser and comparable methods, the amount of sand was slightly lower compared to PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 6 / 16 PLOS ONE Particle-size distribution predicted by infrared spectroscopy Fig 2. Particle size distribution of studied soils in the Canadian textural diagram. https://doi.org/10.1371/journal.pone.0233242.g002 Fig 2. Particle size distribution of studied soils in the Canadian textural diagram. https://doi.org/10.1371/journal.pone.0233242.g002 Fig 2. Spectral data modeling To conduct predictions to details of methodologies and features, the data set was divided into eight subsets (Table 3). Prediction models were run for each subset. In general, prediction accuracies were weakest for the silt fraction and were similar whether data were crude or ilr- 7 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 Fig 3. Distribution of ilr textural variables across methodologies. https://doi.org/10.1371/journal.pone.0233242.g003 Table 2. Comparison of methods (method1 minus method2) using paired t-test and confidence intervals (p 0.05). Target variable Method1 Method2 p.value N [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Sedimentation peroxide ns 38 [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Laser 0 min 46 [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Laser 2 min 106 [Carbon \| Clay,Silt,Sand] Sedimentation 2-pointpoint No peroxide Sedimentationmulti-pointpoint No peroxide 763 [Carbon \| Clay,Silt,Sand] Sedimentation 2-pointpoint peroxide Sedimentation multi-points peroxide ns 227 [Clay \| Silt,Sand] Sedimentation No peroxide Sedimentation peroxide 38 [Clay \| Silt,Sand] Sedimentation No peroxide Laser 0 min 46 [Clay \| Silt,Sand] Sedimentation No peroxide Laser 2 min 106 [Clay \| Silt,Sand] Sedimentation 2-point No peroxide Sedimentation multi-point No peroxide 763 [Clay\| Silt,Sand] Sedimentation2-point peroxide Sedimentation multi-point peroxide 227 [Silt \| Sand] Sedimentation No peroxide Sedimentation peroxide ns 38 [Silt \| Sand] Sedimentation No peroxide Laser 0 min 46 [Silt\| Sand] Sedimentation No peroxide Laser 2 min 106 [Silt \| Sand] Sedimentation 2-point No peroxide Sedimentation multi-point No peroxide 763 [Silt \| Sand] Sedimentation 2-point peroxide Sedimentation multi-point peroxide 227 Sand Sedimentation No peroxide Sieving 746 Sand Sedimentation peroxide Sieving ns 26 Sand Laser 0 min Sieving 8 34 Sand Laser 2 min Sieving 77 Sand Sedimentation multi-point No peroxide Sieving ns 358 Sand Sedimentation multi-point peroxide Sieving 84 ns, ,: Non-significant and significant at the 0.05 and 0.01 levels, respectively. N: Sample size. https://doi.org/10.1371/journal.pone.0233242.t002 PLOS ONE Particle-size distribution predicted by infrared spectroscopy PLOS ONE Particle-size distribution predicted by infrared spectroscopy Fig 3. Distribution of ilr textural variables across methodologies. Fig 3. Distribution of ilr textural variables across methodologies. Fig 3. Distribution of ilr textural variables across methodologies. Table 2. Comparison of methods (method1 minus method2) using paired t-test and confidence intervals (p 0.05). Spectral data modeling Target variable Method1 Method2 p.value N [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Sedimentation peroxide ns 38 [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Laser 0 min 46 [Carbon \| Clay,Silt,Sand] Sedimentation No peroxide Laser 2 min 106 [Carbon \| Clay,Silt,Sand] Sedimentation 2-pointpoint No peroxide Sedimentationmulti-pointpoint No peroxide 763 [Carbon \| Clay,Silt,Sand] Sedimentation 2-pointpoint peroxide Sedimentation multi-points peroxide ns 227 [Clay \| Silt,Sand] Sedimentation No peroxide Sedimentation peroxide 38 [Clay \| Silt,Sand] Sedimentation No peroxide Laser 0 min 46 [Clay \| Silt,Sand] Sedimentation No peroxide Laser 2 min 106 [Clay \| Silt,Sand] Sedimentation 2-point No peroxide Sedimentation multi-point No peroxide 763 [Clay\| Silt,Sand] Sedimentation2-point peroxide Sedimentation multi-point peroxide 227 [Silt \| Sand] Sedimentation No peroxide Sedimentation peroxide ns 38 [Silt \| Sand] Sedimentation No peroxide Laser 0 min 46 [Silt\| Sand] Sedimentation No peroxide Laser 2 min 106 [Silt \| Sand] Sedimentation 2-point No peroxide Sedimentation multi-point No peroxide 763 [Silt \| Sand] Sedimentation 2-point peroxide Sedimentation multi-point peroxide 227 Sand Sedimentation No peroxide Sieving 746 Sand Sedimentation peroxide Sieving ns 26 Sand Laser 0 min Sieving 8 34 Sand Laser 2 min Sieving 77 Sand Sedimentation multi-point No peroxide Sieving ns 358 Sand Sedimentation multi-point peroxide Sieving 84 ns, ,: Non-significant and significant at the 0.05 and 0.01 levels, respectively. N: Sample size. https://doi.org/10.1371/journal.pone.0233242.t002 Table 2. Comparison of methods (method1 minus method2) using paired t-test and confidence intervals (p 0.05). 8 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 PLOS ONE Particle-size distribution predicted by infrared spectroscopy Fig 4. Mean differences (p 0.025) in textural balances between several methodologies against the reference 2-point 2-h sedimentation method without pre-treatment. https://doi.org/10.1371/journal.pone.0233242.g004 Fig 4. Mean differences (p 0.025) in textural balances between several methodologies against the reference 2-point 2-h sedimentation method without pre-treatment. Fig 4. Mean differences (p 0.025) in textural balances between several methodologies against the reference 2-point 2-h sedimentation method without pre-treatment. https://doi.org/10.1371/journal.pone.0233242.g004 https://doi.org/10.1371/journal.pone.0233242.g004 https://doi.org/10.1371/journal.pone.0233242.g004 transformed. As expected, the clay fraction determined by laser in Set1 was poorly related to the NIR spectra (R2 = 0.45–0.64) whether the PSD was expressed as ilr or % (Table 3). Com- bining laser and sedimentation methods in Set2 improved the clay predictions with R2 values of 0.85–0.91. Spectral data modeling In Set3, PSD predictions were more accurate with the 2-point 2-h sedimentation method (R2 = 0.49–0.79) than with the multi-point sedimentation method (R2 = 0.44–0.70). Modeling Set4 showed that NIR was less accurate (R2 = 0.40–0.71) compared to MIR (R2 = 0.45–0.80). Accuracy was higher with NIR-4X (R2 = 0.87–0.95) than with NIR-2X (R2 = 0.69– 0.92) (Set5). Models for Set6 showed that data expressed as ilr or % (R2 = 0.76–0.94) for inter- polated PSD were similarly accurate to the slope and intercept from the relationship between settling time and suspension density (R2 = 0.84–0.92). Several features in Set7 improved model accuracy (R2 = 0.86–0.91) compared to no features at all (R2 = 0.82–0.97). The total carbon content contributed the most to the increase in accuracy, followed by colors and oxalate extracts. Finally, the carbon content showed higher performance with raw data (R2 = 0.63– 0.89) than with ilr-transformed data (R2 = 0.28–0.84). Compared to a model without features (R2 = 0.97), the reconstituted bulk density (R2 = 0.99), color (R2 = 0.96), oxalate (R2 = 0.98) and Mehlich-3 (R2 = 0.98) extracts were similar to the accuracy of the NIR model. For the sand fraction, the NIR model calibrated against the sand-sieving method showed higher accuracy (R2 = 0.66) compared to the sedimentation 2-point 2-h method (R2 = 0.53). PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 Discussion Methodologies. The IR technologies are generally calibrated against a single methodol- ogy. In this paper, several methodologies were IR-tested on comparable soil samples. The present paper presented IR-SGB results across several methodologies and features that are rarely addressed altogether in the literature. Machine learning integrated the available PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 9 / 16 Particle-size distribution predicted by infrared spectroscopy PLOS ONE Table 3. Accuracy, slope, intercept, and number of observations of models. Set Dependent Variables Independent variables Laboratory method for calibration Sand Silt Clay C Sand Silt Clay C Value interpretation N Adjusted R2 RMSE Set1 Ilr 2X Laser 0.80 0.79 0.45 0.28 10.80 10.64 1.32 0.67 - 92 % 0.86 0.80 0.64 0.63 9.11 10.35 1.06 0.48 - Set2 Ilr Lab method, PT, NIR- 2X 2-point 2-h sedimentation + Laser 0.90 0.85 0.80 0.58 9.05 6.94 5.27 0.72 + 485 % 0.89 0.91 0.85 0.71 9.30 7.17 4.21 0.62 ++ Set3 Ilr NIR-2X Multi-point 7-h sedimentation 0.63 0.44 0.70 0.40 16.17 13.22 6.94 0.92 - 667 2-point 2-h sedimenttion 0.76 0.49 0.79 0.36 13.42 11.50 6.60 0.95 - Set4 Ilr MIR 2-point 2-h sedimentation 0.75 0.45 0.80 0.21 14.75 11.34 7.88 0.77 - 222 NIR-2X 0.71 0.40 0.69 0.02 15.78 11.81 9.75 0.86 - Set5 Ilr NIR-4X 2-point 2-h sedimentation 0.94 0.87 0.95 0.85 7.26 6.15 3.62 0.48 ++ 311 NIR-2X 0.83 0.69 0.92 0.76 12.42 9.40 4.78 0.62 + Set6 Ilr PT, NIR-2X 2-point 2-h sedimentation 0.90 0.81 0.93 0.84 8.75 6.98 3.94 0.73 + 860 % 0.91 0.76 0.94 0.89 8.40 7.74 3.84 0.61 + Slope, Intercept 0.90 0.84 0.92 - 8.63 6.34 4.29 - ++ Set7 Ilr All features, NIR-2X 2-point 2-h sedimentation 0.91 0.86 0.90 0.69 9.27 5.86 6.06 0.36 ++ 156 No feature, NIR-2X 0.94 0.82 0.97 0.97 7.78 7.67 3.07 0.71 ++ Carbon, NIR-2X 0.96 0.88 0.97 0.95 6.59 6.34 3.36 0.97 ++ Bulk density, NIR-2X 0.89 0.63 0.94 0.99 10.59 10.84 4.66 0.52 - pH, NIR-2X 0.92 0.78 0.96 0.94 8.84 8.39 3.53 1.06 + Color, NIR-2X 0.94 0.86 0.95 0.96 7.55 6.64 4.10 0.84 ++ Oxalate, NIR-2X 0.95 0.84 0.96 0.98 7.19 7.17 3.71 0.54 ++ Mehlich3, NIR-2X 0.89 0.70 0.93 0.98 10.62 9.83 4.71 0.64 + Set8 % NIR-2X Sand sieving 0.66 - - - 12.15 - - - + 746 2-point 2-h sedimentation 0.53 - - - 14.73 - - - - C: Carbon; PT: Pre-treatments (no peroxide or peroxide); MIR: MIR scores; RMSE: Root mean square error; N: Sample size. : Value interpretation on sand, silt and clay contents. -: Vulnerable estimations; +: Approximated estimations; ++: Research application estimations; +++: Quality control estimations [48,49]. C: Carbon; PT: Pre-treatments (no peroxide or peroxide); MIR: MIR scores; RMSE: Root mean square error; N: Sample size. PLOS ONE : Value interpretation on sand, silt and clay contents. -: Vulnerable estimations; +: Approximated estimations; ++: Research application estimations; +++: Quality control estimations [48,49]. https://doi.org/10.1371/journal.pone.0233242.t003 information on pre-treatments, methods and features to predict soil texture at low cost in routine laboratories. Sedimentation methods. The peroxide pre-treatment tended to increase the clay and silt fractions at the expense of the sand fraction, indicating that clay- and silt-size particles were freed from the clay- or silt-organic matter complexes in micro-aggregates [50]. While metals such as Fe are sequestered by soil organic matter [51] or associated with it [52], iron may be freed from organic matter using hydrogen peroxide to form reactive iron hydroxide [53] that may, conversely, positively impact soil aggregation [54]. Peroxide-treated soils may thus require additional dispersion prior to sedimentation [55]. Nevertheless, the 2-point 2-h sedi- mentationwith and without peroxide pre-treatments showed comparable results or minor dif- ferences for the [clay \| silt,sand] and [silt \| sand] balances. Laser method. The Mie theory considers soil particles to be spherical (Malvern Instru- ments Ltd., 2007). Parameters such as the refractive index of the medium, the real or imaginary refractive index and the density could be included to improve accuracy. The 2-min sonicated laser method and the sand sieving method returned closed results. The 40 kHz sonication for a duration of 2 min appeared to be suitable for sandy soils [56]. Variants include 36 kHz for 3 10 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 PLOS ONE Particle-size distribution predicted by infrared spectroscopy min [57] and 30 kHz for 30 min [2]. Although fragile quartz grains may be broken using ultra- sonication [58], the [silt \| sand] balance for the sedimentation method showed results close to the 2-min sonicated laser method. According to Storti and Balsamo (2010) [59], the high-strength materials are not as affected by the procedures as the low-strength materials. Using water as the dispersing agent and 1750- , 700- rpm for the pump and stirrer speeds, Sochań et al. (2012) [60] obtained R2 values of between 0.67 and 0.95 for high-strength materials in silt loamy to sandy soils, values close to our raw data results (R2 = 0.64–0.86). For clayed soils, a surfactant or solvent may replace tap water [61]. As small particles increase in number and combine into sand-sized aggregates, pre- treatments may be required and adjusted to soil specificities. Spectral data modeling In this study, we used subsets 92 samples and GBM as the machine learning method for par- ticle-size prediction compared to suggested subsets 130 samples for cLHS and FCMS [62]. Here, only Set1 for laser samples did not meet that criterion. The Boosted Regression Trees is another suitable ML method for complex predictions [41] and is relatively fast compared to random forest and neural network algorithms. Model accuracies in validation were in the range reported in the literature where R2 values have been found to vary between 0.46 and 0.94 for clay [10,63,64] and between 0.53 and 0.82 for sand. More generally, the R2 values may vary widely between 0.05 and 0.84 [65,66]. Prediction accuracy. Because the 2-point 2-h sedimentation with and without pre-treat- ment showed comparable results, they could be combined for IRS calibration with method and treatment information inputs. Hence results of soil surveys could provide a large database for IRS calibration purposes. Laser methods produced results with lower accuracy compared to the sedimentation meth- ods, due to underestimation of the clay fraction. The [silt \| sand] balance from the laser method, however, was close to that of the 2-point 2-h sedimentation method, despite higher silt and sand percentages for the laser method, indicating advantage for log-ratioing. Higher accuracies from the laser method were obtained by Blott and Pye (2006) [67] across a wide range of soils, sediments and powders. Zobeck (2004) [68] related results from a LS-230 laser diffraction particle size analyzer to those of the pipette method and obtained R2 values of 0.97, 0.99, and 0.99 for the < 2-, < 50-, and < 100 –μm in non-calcareous soils, using a shape factor of 0.2 compared to the default 1.0 shape factor for the laser method. Nevertheless, we found that the laser method did not produce results as consistent as the sedimentation method for clay predictions by IRS, indicating regional, soil-specific, calibration. Mehlich-3 extracts, reconstituted bulk density and pH did not improve prediction accuracy. Soil features such as total carbon content, colors and amorphous materials (oxalate extracts) increased model prediction accuracy. Prediction accuracy of carbon content could perform with features as reconstituted bulk density as well as Mehlich-3, colors and oxalate extracts. PLOS ONE There is no standard procedure to disperse soil samples for the laser method because pre-treatment is soil specific. PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 Supporting information S1 Table. Slope, intercept, and number of observations of models. (DOCX) S2 Table. Accuracy of GBM and neural network models using k-fold cross validation (k = 10). (DOCX) S1 File. (PDF) S1 Table. Slope, intercept, and number of observations of models. (DOCX) S1 Table. Slope, intercept, and number of observations of models. (DOCX) S2 Table. Accuracy of GBM and neural network models using k-fold cross validation (k = 10). (DOCX) S1 File. (PDF) Spectral data modeling Using suspension density function parameters instead of arbitrary settling times did not increase the accuracy of PSD predictions but provided a uniform base to run NIR models as various settling times have direct impact on predictions. Then, in Set3, we observed that 0.67-min and 2-h settling times were more accurate than 0.75-min and 7-h periods for sand and clay NIR predictions. In the present study, MIR spectra were more accurate than NIR for sand, silt and clay deter- minations. The NIR method can provide accurate prediction for clay as it has been also found to be accurate for cation exchange capacity (CEC) with R2 values of 0.82 [41] and 0.81 [63]. On PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021 11 / 16 PLOS ONE Particle-size distribution predicted by infrared spectroscopy the same direction, Viscarra Rossel et al. (2006) [9] concluded that MIR was more suitable than NIR for texture and carbon determination, due to higher incidence of spectral bands combined with higher intensity and specificity of the signal compared to NIR. To further sup- port NIR calibration and model accuracy, the R-coded GBM machine learning model used in the present study came across several soil textural classes, carbon contents and features that have not been addressed simultaneously in past research. Conclusions In this paper, IRS inaccurately predicted the PSD’s clay fraction using laser methods. However, IRS accurately predicted PSD against sedimentation and sieving methods after adding soil fea- tures such as color, total carbon content and concentration of amorphous materials related to soil genesis and classification. Soil pre-treatments and the need for dispersing agents could be adjusted to the nature and concentration of binding agents for silt-, clay-size particles and fine particles adhering to sand particles. Combined with method and treatment as features, post- screening total carbon content and color routinely determined in service laboratories can improve IRS accuracy for mineral soils. Features as reconstituted bulk density and Mehlich-3 extracts could be added as features for higher-C soils. The GBM returned similar results whether particle-size data were analyzed as raw pre- determined settling times (percentages) or as ilr-transformed percentages. The GBM returned similar accuracies using the slope and intercept of the log-log relationship between settling time and suspension density, ilr-transformed percentages or raw percentages. The MIR and NIR 4X gain methods performed better than the NIR 2X gain method. However, additional features increased NIR 2X predictability. Modeling the log-log relationship between settling time and suspension density provided greater flexibility in the choice of soil-specific settling times. The GBM model proved to be a powerful tool to process the results of several analytical methods used worldwide to determine soil grain-size distribution [8]. More methodologies could be included in the future such as the pipette methods, and chemical and sonication methods to disperse microaggregate. Author Contributions Author Contributions Conceptualization: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Formal analysis: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Funding acquisition: Le´on Etienne Parent. Investigation: Elizabeth Jeanne Parent, Le´on Etienne Parent. Methodology: Elizabeth Jeanne Parent, Serge-E´tienne Parent, Le´on Etienne Parent. Project administration: Le´on Etienne Parent. Resources: Le´on Etienne Parent. Software: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Validation: Serge-E´tienne Parent. Visualization: Le´on Etienne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – review & editing: Le´on Etienne Parent. Author Contributions Conceptualization: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Formal analysis: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Funding acquisition: Le´on Etienne Parent. Investigation: Elizabeth Jeanne Parent, Le´on Etienne Parent. Methodology: Elizabeth Jeanne Parent, Serge-E´tienne Parent, Le´on Etienne Parent. Project administration: Le´on Etienne Parent. Resources: Le´on Etienne Parent. Software: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Validation: Serge-E´tienne Parent. Visualization: Le´on Etienne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – review & editing: Le´on Etienne Parent. Author Contributions Conceptualization: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Formal analysis: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Funding acquisition: Le´on Etienne Parent. Investigation: Elizabeth Jeanne Parent, Le´on Etienne Parent. Methodology: Elizabeth Jeanne Parent, Serge-E´tienne Parent, Le´on Etienne Parent. Project administration: Le´on Etienne Parent. Resources: Le´on Etienne Parent. Software: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Validation: Serge-E´tienne Parent. Visualization: Le´on Etienne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – review & editing: Le´on Etienne Parent. Conceptualization: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Funding acquisition: Le´on Etienne Parent. Investigation: Elizabeth Jeanne Parent, Le´on Etienne Parent. Methodology: Elizabeth Jeanne Parent, Serge-E´tienne Parent, Le´on Etienne Parent. Project administration: Le´on Etienne Parent. Methodology: Elizabeth Jeanne Parent, Serge-E´tienne Parent, Le´on Etienne Parent. Project administration: Le´on Etienne Parent. Software: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Software: Elizabeth Jeanne Parent, Serge-E´tienne Parent. Validation: Serge-E´tienne Parent. Visualization: Le´on Etienne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – original draft: Elizabeth Jeanne Parent. Writing – review & editing: Le´on Etienne Parent. 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Particle size distribution analysis of sand-sized particles by laser diffraction: an experi- mental investigation of instrument sensitivity and the effects of particle shape. Sedimentology. 2006; 53 (3):671–85. References 67. Blott SJ, Pye K. Particle size distribution analysis of sand-sized particles by laser diffraction: an experi- mental investigation of instrument sensitivity and the effects of particle shape. Sedimentology. 2006; 53 (3):671–85. 68. Zobeck T. M. RAPID SOIL PARTICLE SIZE ANALYSES USING LASER DIFFRACTION. Appl Eng Agric. 2004; 20(5):633–9. 16 / 16 PLOS ONE \| https://doi.org/10.1371/journal.pone.0233242 July 20, 2021	5,101
W4387619361.txt_1	Open-Science-Pile	Open Science	Various open science	2,023	Design, Development and Performance Evaluation of Semi Automated Citrus Juice Extractor Machine	None	English	Spoken	3,941	6,497	Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 Design, Development and Performance Evaluation of Semi Automated Citrus Juice Extractor Machine 1 Ugwu Benedict Nnamdi1, Ani John Ndubuisi2 and Ilo Paulinus Chukwudi3 Department of Mechanical and Production Engineering, Enugu State University of Science and Technology, Agbani Enugu State, Nigeria 2 Department of Mechanical and Production Engineering, Enugu State University of Science and Technology, Agbani Enugu State, Nigeria 3 Department of Mechanical and Production Engineering, Enugu State University of Science and Technology, Agbani Enugu State, Nigeria 1 Corresponding Author: benedict.ugwu@esut.edu.ng Received: 01-09-2023 Revised: 15-09-2023 Accepted: 29-09-2023 ABSTRACT Citrus juice extractor machine has been design, fabricated and can be found in numerous studies at different areas. These citrus juice extractor machine is a semi automated one that operates from the entrance of citrus into the hopper basket till the filteration of the juice for consumption. The operational procedure for the extraction of juice starts from the stacking of neatly cleaned citrus of same size in the basket position ontop of the extractor machine which in turn passes through the hopper to the extraction chamber where the oranges are cut into two equal halves for the twin knaggy ball shearing then the pulps are dropped through the remaning collector to the residue collection bin while the extracted juice flows through the seize to a container for consumption. The optimal operating speed of the juice extraction machine was found to be 15rpm, while the optimum feed was found to be F1 (2.5 kg/min). The average juice extraction efficiency at optimum speed (S) and feed rate (F) was 64%; juice yield at optimum Speed and Feed was 60%; juice extraction losses was 35% at optimum Speed and Feed. Keywords: remaining collector, top basket hopper, juice extraction chamber, knaggy ball I. INTRODUCTION Citrus plants are grown and cultivated in most parts of world with north eastern India as the originators [1]. The world production of oranges was led by Brazil, China, India, EU, USA, Mexico, Egypt as the largest producers in 2019-2020 projected as 76 million metric tons (84 million short tons), [2]. The fiscal Year 2020/2021 showed Brazil as the leading universal orange producer, with production volume of 14.7 million metric tons against the overall production volume of fresh oranges summed to about 47.45 million metric tons which is expected to increase to 47.8 million metric tons in 2022/2023 [3], The Year 2022/2023 projection shows that the top ten citrus producing counties are Brazil, China, European Union, Mexico, United States, Egypt, Turkey, South Africa, Morocco and Vietnam [4]. Due 140 countries were reported to be involved in citrus production [5]. The fruit is in abundant during its harvesting period and always in short supply and expensive during its off season with about 30% loss of the fresh fruit [6]. Attempts to store these fruits in its fresh and natural form have been fruitless as a result of its perishable nature evident in its elevated moisture content, deplorable post-harvest management and marketing approach [7]. Research conducted on the physical properties of orange using reasonable quantities that is above hundred revealed that the mass of the orange fruit ranges between 160 - 170g [8 – 9]. Orange in its natural form has shelf life between 2-10 days but it can still be improved upon by storing in a proscribed temperature or pressure, procession into more established products like juices, jellies and jam, refrigeration, packaging in plastic films, use of food surface coatings, chemical treatments and irradiation [10-11]. However, the cost of the fruit is so little as to compensate for the cost of using this method in countries like Nigeria [12]. Fruit processing into juice is adjudged to be an improved way of storage, preservation and value addition. Juice extraction and separation thus becomes a new market opportunity for modern consumption of fruit products. Orange extraction evolution commenced by hand extraction of the juice which is quite dawdling, tiresome and unhealthy; while the demand for juice consumption increases by machine extractor existence [13]. The juice extraction machines have the benefits of less time usage, enhanced efficiency, improve juice yield and less waste and junk [14]. Locally fabricated juice extractors have existed but have its shortcoming in terms of limited output [15]. These juice extractor is an improved extractorl machine which utilizes https://asejar.singhpublication.com 58 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 the pressing means to extract juices from some citrus fruit [16]. Citrus (oranges) and citrus yield have enriched nutritional contents; that are rich and inexpensive sources of vitamins (particularly vitamin C), minerals and dietary fiber required for healthy living [17]. Fruit juices are the extracted tissues fluid content of the fruit or the extraction of fermentable fluids of the fruit for the use of human beings, through mechanical process from entirely nutritional fruits [10]. Agricultural mechanization is a sure way to boosting home-grown design, invention and manufacture of most machines and equipment required for technology advancement [18]. Although, different size and shape of citrus fruits exist, but there is amazingly little or no basic disparity in the extraction process of juice from them [16]. To address this problem there is an urgent need to design and fabrication an automated fruit juice extractor appropriate for this range of fruits for commercial needs of the farmers and the majority of our teeming populace [17]. This developed juice extractor machine will be a simple, reasonably priced machine that is eco-friendly, energy efficient and flexible in the processing of citrus fruits. This machine outcome will provide local substitute to imported brands of processed fruit juices machines and also reduction of wastage during the citrus harvest seasons. Figure 1: Solid work Design of Juice Extractor Machine Figure 2: Exploded view of the Extraction Chamber https://asejar.singhpublication.com 59 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 II. Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 DESIGN CONSIDERATIONS AND CALCULATIONS The machine is fabricated such that it will remain stable while in operation. It consists of electric motor, hopper, orange remaining collectors, shafts, Knaggy rotary balls, gears, waste outlet, juice outlet and main frame. The juice extractor design works on three principles: convey, slice and squeeze. The machine components are held and supported by a rectangular shape main frame formed to give a compact design and stronger outlook. The hopper is mounted on top of the extraction chamber. This feed hopper is circular in shape and attached to a shaft to enable it feed the oranges into extraction chamber of the machine. perforation placed below the two knaggy balls at the extraction chamber. The already squeezed orange halves are trashed through the pulp outlet as residual waste on both sides of the machine. The diagram in figures 1- 4 explains the operational sequence of the orange juice extractor machine. An orange juice extractor has many components, each of which contributes to the overall performance and ultimately affects the operational results. Good choice of material and well-designed components that are complementary to each other will keep the overall performance within an acceptable limit. Selecting suitable materials for the various machine components is one of the most important tasks in the design.The decision of what materials to be used was made before the actual dimensions of the parts were determined since load and stress calculations are based on the properties of the materials. Along with the choice of a material, the process which the material would be formed into the machine was also considered. Although, a given material may have the required properties for satisfactory service but it is also important to decide on the forming/shaping process of the machine part as the two combined determines the cost of the part or machine and represents the maximum value of money to be expended. As important as the stress and deflection of mechanical parts are, the selection of a material is not only based upon these factors alone. Other factors that are considered for a satisfactory material selection are weight, thermal properties, resistance to corrosion, resistance to wear, availability, cost, appearance, safety and ease of fabrication. The various properties and functions of each component in the machine determine the choice of material for its construction. For the machine housing, stainless steel is used.The whole machine can be described in three compartments namely: carrier (hopper), Extracting chamber and Frame as shown in solid model of figure 1. The main function of the frame in the design of the extractor machine is its provision of the appropriate relative position of the units and component parts mounted on it over all working conditions and period of service. The strength of the machine is the second required consideration made in the design of the extractor machine. The frame structure was design to accommodate different weights of components mounted on it, and it is made of mild steel with overall dimension of 393mm × 220mm × 581mm. The hopper (item A) in figure 1 is an essential part of the machine that guides and retains fruit to be processed. It acts as a container and at the same time helps in the gradual introduction of the orange fruit into the juice-extracting chamber. The circular shaped hopper is located on top of the housing. Galvanized metal sheet was used for the “hopper” all the same stainless steel or alloys of aluminum can substitute. The juice extraction is carried out as shown in figure 4, extraction chamber. It consists of the orange collector and rotary balls which are arrange on the shaft along its length alternatively. It also consists of the knife that cut the oranges in two equal halves. The knife is situated at the center of the two orange collectors. Furthermore, the juice collector and sieve are also located in this compartment. The sieve filters the juice while the collector collects it into the cup. For better filtering it is expected that sieves should be changed after each day of operation. The other components of the extractor include: Shafts, Gears (spur gears) and chain drivers, Electric motor which rotates the shaft connected to the knaggy balls, Bearings, Sieve for juice filtration after extractor, Waste separator for separating the chaff from the juice being extracted, Switch: For turning the machine ON/OFF Rotary balls for pressing and extracting the orange juice. Shaft is a rotating member of the machine, see figure 1 (item D). Mild steel or galvanized steel can serve as the material for shaft. The press components (orange collectors and rotary balls) which are of the same material (Teflon material or wood were mounted on the shaft, set alternatively and evenly at equal intervals along its length. The shafts are seven out of which six carries spur gears of the same number of teeth while the seventh carries bevel gear for the hopper which has lesser number of teeth. The bevel gear engages with another bevel gear attached on one horizontal shaft. The six gears are arranged in parallel, with three gears on each side. Two orange collectors and two rotary balls are driven by two other gears that transmit torque to the gears on the shaft of orange collector and rotary balls Rotary balls are made up of Teflon material but wood can also be used. Each consists of three balls on a single axis of 1200 from each other and radius of 57 mm balls attached at the ends. The length and size of the rotary balls is tantamount to the size of the compressing chamber (collector). For effective pressing two pairs of rotary balls were developed. The weight of the hopper, shaft, gears, housing structure are been carried by the machine base and support. The orange collector is a circular shaped object, constructed with a Teflon material or wood; it is two in number with each having three doom shaped recesses that collects the orange. The collector has a height of 100 mm and radius of 73 mm https://asejar.singhpublication.com 60 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 and is been rotated by a shaft attached to one of the six gears. A gear is a toothed wheel that works by engaging another toothed system arranged to change the speed or direction of transmitted motion. Figure 3: Exploded view of the Juice Extractor Machine Figure 3 shows seven spur gears G1, G2, G3…G7 mounted on seven different shafts. Gear G1 rotates at 1 revolution in 4 seconds (15rpm). Where, N1 = Np = 15 teeth, N2 = Ng = 45 teeth, N3 = Ng = 45 teeth, P = 1/3hp = 1.2KW Tooth pressure angle (Ø) is 200 full depth involutes gear of steel material. The angular speed of the gear: (1) (2) Where: TP = Pinion Torque (Nm), P= Power, hp (KW); NP = Pinion speed, rpm. Since, NP = 3 revolutions per 12seconds = 15 rpm Motor power rating (P) = But, 1hp = 0.746 KW TP = 19.8 Nm https://asejar.singhpublication.com 61 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 2.1 Gear Ratio The desired gear ratio is determined by the designer from the given input speed and expected output speed. It is the ratio of rotational speed of input gear to that of the output gear and can also be referred to as velocity ratio. (3) Where, NP = Number of teeth on pinion, Ng = Number of teeth on gear, DP = pitch diameter of pinion, Dg = Pitch Diameter of gear. Therefore, Where, Ng = 45, NP = 15 But (4) Where, Tf = output torque, Ti = input torque, Vr = Velocity ratio Since we have Pinion torque (TP ) = 19.8 Nm, Ti = Input torque =?, Vr = Velocity ratio = 3 Hence, Tf = 19.8 x 3 = 59.4 Nm 2.2 Determination of Shaft Diameter on the Basis of Strength The basic considerations in the design of shaft strength, are as follows:The maximum shear stress, τmax, of the shaft is calculated using equation (5) from [19] as shown below, √ (5) Where, , d = Shaft diameter, , , Bending moment is insignificant in this case. Kt is equal to 1.0 for rotating shaft with steadily applied loads. Then equation (5) becomes, (6) } √{ Therefore, Assuming, (7) , T = 10.2N.m From equation (6), √{ } 2.3 Bearing Design For shafts connected by gears in rotating machines without any impact, the load factor, „K‟ assumes any value ranging from 1.1 – 1.5. The desired Life rating „L‟ of the bearing is determined from the equation: ( ) (8) Where, L = Rating of bearing, C= dynamic capacity in Newton, P = equivalent load = 62.98N K = 3, for ball bearings. Assuming 90% of HTE bearing life rating will reach thirty (30) million revolutions before failure, such that L = 30, Then, equation (8) becomes https://asejar.singhpublication.com 62 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Therefore, Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 , √ 2.4 Evaluation of Juice Extractor Performance The evaluation of the electric powered juice extractor performance was performed after the fabrication for the purposes of establishing the optimum juice extracting parameters. The extracted parameters include: 2.4.1 Operating Factors The five levels extraction speed (S) inludes (S 1 = 225, S2 = 150, S3 = 95, S4 = 60 and S5 = 15rpm) and corresponding three levels feed rate (F) as (F1 = 2.5, F2 = 3.0, and F3 = 3.5 kg/min). 2.4.2. Performance Parameters The following are the performance parameters juice yield, JY (%), juice extraction efficiency, EF (%) and extraction losses, EL (%). The gear arrangement aided in obtaining different extraction speeds during the performance test. Using equations 9, 10 and 11 as proposed by [20] and verified in [21]and [22]. The following juice yield, extraction efficiency and extraction loss were calculated as follows: ( ) ( ( (9) ) x 100 (10) ) (11) Where: MJE = Juice extracted mass (kg), MRW = Residual waste/dry pulp mass (kg), MFS = Feed sample mass (kg), JY = Juice yield (%), Eeff = Efficiency of extraction (%), EL = Loss of extraction (%). The juice constant was obtained from the ratio of sum of masses of juice extracted and juice in chaff to the mass of fruit fed in. ( ) (12) Where: x = Juice constant of fruit (decimal), MJC = Mass of Juice in the chaff, The juice constant (x) for orange were determined to be within the range of 0.54 and 0.68, and applied in equation (13) to compute the juice extraction efficiency of the orange as shown below: For example, From Table 3 and at feed rate, F1 = ( ) = 2.5kg/min (13) 2.5 Machine Performance Results https://asejar.singhpublication.com 63 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. RESULTS AND DISCUSSION The Tables 1 and 2 with Figures 4, 5 and 6. Explain the evaluation test results carried out on the juice extraction machine. 3.1 Juice Yield The result in table 1 illustrates the orange juice extractor optimum feed rate as F 3 (3.5 kg/min). More juice yield was obtained from orange fruits because there are sufficiently juicy in nature. Consequently, more juice yield was extracted at each operating speed of the fabricated electric powered juice extractor than was obtained from manually operated extractor. Broadly, the overall juice yield of these machine decreases as the juice extractor machine operating speed increases and vice-versa (Figure 4). As the speed of operation increases, there is a significant reduction in juice extraction from the oranges. The losses may be due to vibrational high operating speeds which causes splashing of juice around the walls of the xtractor, thus reducing its yield. Hence, to obtain high juice yield a controlled speed should be sustained when operating the machine for an optimal juice extraction. 3.2 Extraction Effficiency Figure 5 showed the orange juice extraction efficiency of the machine. The extraction efficiency of machine rose to a maximum value of 70% at optimum operating speed of 15rpm and optimum feed rate F3 (3.5 kg/min), and then declines at speeds beyond 15rpm. Fruits were not completely squeezed at high speeds which thn caused the reduction in the amount of juice extracts compared to when it was tested at a slow and steady speeds. Thus, the performance evaluation tests proved the optimum extraction efficiency of the electric powered juice extractor machine to relies on the fruit nature and the extraction speed of the machine to produce the juice extracts . The rotary balls have a major responsibility to play since the sizes of orange fruit that passes down to the collectors where the fruit juice were finally extracted is considered as prime. The roughness rises as speed increases while there is a decline in extraction time as the extraction speed rises. IV. CONCLUSION The juice extraction machine was successfully designed, neatly constructed, assembled judiciously and tested. The optimum operating speed of the juice extraction machine was found to be 15rpm, while the optimum feed was found to be F1 (2.5 kg/min). The average juice extraction efficiency at optimum speed (S) and feed (F) was 64%; juice yield at optimum S and F was 60%; juice extraction losses was 35% at optimum S and F. The juice extractor machine cost about N125, 000, which makes it affordable for both small and medium-scale farmers in the country. Conflicts of Interest The authors wish to declare no conflict of interest. https://asejar.singhpublication.com 66 \| P a g e Applied Science and Engineering Journal for Advanced Research ISSN (Online): 2583-2468 Volume-2 Issue-5 \|\| September 2023 \|\| PP. 58-67 Peer Reviewed and Refereed Journal DOI: 10.54741/asejar.2.5.9 REFERENCES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. L. G. Albrigo, L. L. Stelinski, & L. W. Timmer. (2019). Citrus. (2nd ed.), CABI Publishers. https://doi.org/10.1079/9781845938154.0000. Lloyd, P. J., & Vautier, K. M. (1999). Chapter 10: The world trade organization. in Promoting Competition in Global Markets. 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https://openalex.org/W3130787411	OpenAlex	Open Science	CC-By	2,021	Stochastic Ordering of Complexoform Protein Assembly by Genetic Circuits	Mikkel H. Jensen	English	Spoken	11,449	18,166	Editor: James R. Faeder, University of Pittsburgh, UNITED STATES Editor: James R. Faeder, University of Pittsburgh, UNITED STATES Received: January 21, 2020 Accepted: May 28, 2020 Published: June 29, 2020 Peer Review History: PLOS recognizes the benefits of transparency in the peer review process; therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. The editorial history of this article is available here: https://doi.org/10.1371/journal.pcbi.1007997 Copyright: © 2020 Jensen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. PLOS COMPUTATIONAL BIOLOGY PLOS COMPUTATIONAL BIOLOGY RESEARCH ARTICLE Stochastic ordering of complexoform protein assembly by genetic circuits Mikkel Herholdt JensenID1, Eliza J. MorrisID1, Hai TranID2, Michael A. NashID3,4, Cheemeng TanID5 1 Department of Physics and Astronomy, California State University, Sacramento, California, United States of America, 2 Department of Chemistry, California State University, Sacramento, California, United States of America, 3 Department of Chemistry, University of Basel, Basel, Switzerland, 4 Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland, 5 Department of Biomedical Engineering, University of California, Davis, California, United States of America a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 * mikkel.jensen@csus.edu (MHJ); cmtan@ucdavis.edu (CT) * mikkel.jensen@csus.edu (MHJ); cmtan@ucdavis.edu (CT) Abstract Top-down proteomics has enabled the elucidation of heterogeneous protein complexes with different cofactors, post-translational modifications, and protein membership. This hetero- geneity is believed to play a previously unknown role in cellular processes. The different molecular forms of a protein complex have come to be called “complex isoform” or “com- plexoform”. Despite the elucidation of the complexoform, it remains unclear how and whether cellular circuits control the distribution of a complexoform. To help address this issue, we first simulate a generic three-protein complexoform to reveal the control of its dis- tribution by the timing of gene transcription, mRNA translation, and protein transport. Over- all, we ran 265 computational experiments: each averaged over 1,000 stochastic simulations. Based on the experiments, we show that genes arranged in a single operon, a cascade, or as two operons all give rise to the different protein composition of complexoform because of timing differences in protein-synthesis order. We also show that changes in the kinetics of expression, protein transport, or protein binding dramatically alter the distribution of the complexoform. Furthermore, both stochastic and transient kinetics control the assem- bly of the complexoform when the expression and assembly occur concurrently. We test our model against the biological cellulosome system. With biologically relevant rates, we find that the genetic circuitry controls the average final complexoform assembly and the variation in the assembly structure. Our results highlight the importance of both the genetic circuit architecture and kinetics in determining the distribution of a complexoform. Our work has a broad impact on our understanding of non-equilibrium processes in both living and synthetic biological systems. OPEN ACCESS Citation: Jensen MH, Morris EJ, Tran H, Nash MA, Tan C (2020) Stochastic ordering of complexoform protein assembly by genetic circuits. PLoS Comput Biol 16(6): e1007997. https://doi.org/10.1371/ journal.pcbi.1007997 Citation: Jensen MH, Morris EJ, Tran H, Nash MA, Tan C (2020) Stochastic ordering of complexoform protein assembly by genetic circuits. PLoS Comput Biol 16(6): e1007997. https://doi.org/10.1371/ journal.pcbi.1007997 Introduction Proteins are synthesized in specific orders to assemble large protein complexes, such as micro- tubule, proteasome, ribosomes, and cellulosome. These protein complexes are assembled both inside and outside cells through the coordination of gene expression, protein transport, and binding processes. Prior work has been assuming that protein complexes have a homogeneous composition of protein members. Yet, recent top-down proteomics shows that protein com- plexes can compose of different cofactors, post-translational modifications, and protein mem- bership [1–3]. The different molecular forms of a protein complex have come to be called the complex isoforms or complexoforms [1]. For example, recent work shows that the yeast homo- tetrameric FBP1 complex can co-exist with 0 to 4 phosphorylated amino acids [4]. Bacteria cel- lulosomes are also found to exist in heterogeneous compositions [5–11]. Furthermore, a recent computational study [12] has investigated the formation of protein complexes using existing data on protein-protein interaction networks. This prior work shows that the compo- sition of a protein complex can drift over time even when the simulation starts from the same initial condition. The work suggests that other cellular mechanisms must exist to prevent the compositional drift of some protein complexes. While the elucidation of complexoforms is now possible, finding the mechanisms that con- trol the distribution of complexoforms is challenging. There are two competing paradigms for understanding the formation of complexoforms. A classical but unproven paradigm assumes that temporal ordering of protein synthesis is essential for accurate assembly of protein com- plexes. Yet, most biochemical studies mix proteins in a test tube to study protein complex assembly, neglecting the underlying cellular networks that control synthesis and transport of the proteins. This approach suggests a second, contradicting paradigm: the underlying cellular networks are not essential for the assembly of quaternary protein complexes. One way to resolve the contradiction is to examine genome sequences for the spatial arrangement of genes in the genome: if genes are arranged in specific patterns that correlate with their synthesis order or the assembly sequence of protein complexes, then the underlying gene networks may be essential for the protein assembly. Indeed, a study of the human genome shows that housekeeping genes expressed in most tissues show strong clustering [13]. The data suggest that the ordering of protein synthesis may be conserved for critical protein complexes [14]. PLOS COMPUTATIONAL BIOLOGY PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Physics and Astronomy Chien Hu Research Award Program, the Edwin L. Iloff Endowment, and NSF (1808237). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. further research into the importance and the underlying mechanisms of the variation. Here, we study the arrangement of genes as a key factor that modulates the variation of protein complexes. We run computer simulations to investigate how various reaction parameters control the variation of a protein complex. Finally, we extend our framework to study the variation of an enzymatic complex that digests cellulose. Our work has a broad impact on the understanding of protein-complex assembly and set up the new research direction about the variation of protein complexes. Competing interests: The authors have declared that no competing interests exist. Competing interests: The authors have declared that no competing interests exist. Data Availability Statement: All data are included in S1 Data. Data Availability Statement: All data are included in S1 Data. Multiple protein subunits can come together to form protein complexes that play critical functional roles in a cell. Recent advancement in measurement technologies has revealed tremendous variation in the members of protein complexes. The recent results motivate Funding: The work is supported by Human Frontier Science Program (FR) (RGY0080/2015), California State University, Sacramento, Department of 1 / 18 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 Introduction For non-critical protein complexes, either each cell-type evolves its protein synthesis order to achieve unique complexoforms, or the protein synthesis order is not an important factor. A central question currently challenging the field is, therefore, what underlying rules govern protein-synthesis or assembly order, and the limit of the rules in controlling the distribution of a complexoform. Here, we use stochastic computational simulations to model protein expres- sion and assembly from the bottom-up. We first simulate a three-protein model system to investigate how complexoform composition is affected by the timing of gene transcription, PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 2 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits mRNA translation, and protein transport, as well as the gene circuit architecture. Due to the small number of chemical agents and proteins often involved in protein assembly processes, ordinary equilibrium statistical mechanics approaches are not expected to adequately describe the eventual protein assembly outcome. We show that genetic circuits, physical transport, and binding kinetic rates all modulate the distribution of complexoform. Comparison of the sto- chastic simulations to deterministic solutions reveal two distinct kinetic regimes: a slow equi- librium regime in which the average structure is well predicted by equilibrium statistical mechanics, and a fast nonequilibrium regime in which small-number statistics leads to struc- tures far from equilibrium predictions. This work points to new and previously unappreciated mechanisms of modulating the distribution of a complexoform. Our work also highlights the importance of both gene circuit architecture and nonequilibrium stochasticity in regulating the formation of complexoforms. Our work provides a possible resolution of the gene-circuit vs. protein-only paradigms: gene-circuit-based regulation of complexoform distribution may be important only under the select kinetic regime and cellular context. https://doi.org/10.1371/journal.pcbi.1007997.g001 The type of genetic circuit modulates complexoform assembly When a small number of molecules are involved in each process, the exact ratio of each is stochastically determined. To establish the average distribution of complexoform, simulations are run 1,000 times for each genetic cir- cuit and each set of parameters. This number of simulations is found to be sufficient to reach a reproducible average to within an uncertainty of 1 percentage point of XX, XY, and YY. We set up four common types of genetic circuits expressing two proteins, X and Y. The series uncoupled circuit expresses the two proteins from the same operon, but the translation occurs at two separate ribosome binding sites. The series coupled circuit expresses the two pro- teins from the same operon and the translation occurs through a single ribosome binding site. The cascade circuit expresses the two proteins from two separate promoters linked in a cas- cade. Finally, the parallel circuit expresses the two proteins independently from two identical promoters. We first investigate the role of the specific genetic circuit architecture in determining the protein assembly by assuming that the rate constants are identical. This assumption helps to isolate the effect of gene circuit architecture on the formation of complexoform. It will be relaxed in subsequent simulations. Each genetic circuit modulates the distribution of the com- plexoforms differently (Fig 2). For the cascade, series uncoupled, and series coupled circuits, the final complexoforms are predominantly XX. This trend is most evident for the series cou- pled circuit (84% XX, 15% XY, 1% YY) and the cascade circuit (86% XX, 10% XY, 4% YY). The results arise because these circuits produce the mRNA for X or protein X first. For the series uncoupled circuit, protein X is less prevalent in the final complexoform, but still exceeds pro- tein Y (45% XX, 38% XY, 17% YY). The parallel circuit stochastically produces similar amounts of X and Y in the final complexoform (45% XX, 12% XY, 43% YY). These results indicate that even with identical rate constants and initial concentrations of all the reactants, the genetic cir- cuit architecture can significantly modulate the final distribution of the complexoform. Fig 2. Genetic circuit architecture modulates complexoform assembly. The type of genetic circuit modulates complexoform assembly Protein expression and assembly typically involve a relatively small number of each type of molecule (<1000 per type of protein [15]). In such cases, equilibrium statistical mechanics may not accurately describe the process of protein expression and assembly, since the ensem- ble of reactants is insufficiently small. Thus, the process becomes inherently stochastic in nature. Here, we use a stochastic model to examine the heterogeneity in protein expression and assembly. Our model system consists of a genetic circuit expressing two types of proteins, denoted X and Y. The proteins are exported to a sub-cellular location and then bind to scaffold proteins, each consisting of two docking sites. This model system allows us to investigate the role of tim- ing and genetic circuit architecture in modulating the protein assembly, irrespective of the exact number of types of proteins and binding sites. In this model, the binding between the proteins creates a protein complex (Fig 1). The proteins compete for 10 scaffold proteins. The Fig 1. Illustration of the complexoform assembly model system. Simulated processes and rates are indicated by arrows and italicized text. In the model, genetic circuitry coding for either protein X or protein Y is initiated by a promoter and transcribed to mRNA. Translational machinery then produces proteins X and Y, which are exported and compete for a limited number of scaffold proteins, each with two docking sites. The simulation also incorporates a rate of loss of mRNA and external protein. Generic simulations are carried out with 10 external scaffold proteins (i.e., 20 docking sites). Fig 1. Illustration of the complexoform assembly model system. Simulated processes and rates are indicated by arrows and italicized text. In the model, genetic circuitry coding for either protein X or protein Y is initiated by a promoter and transcribed to mRNA. Translational machinery then produces proteins X and Y, which are exported and compete for a limited number of scaffold proteins, each with two docking sites. The simulation also incorporates a rate of loss of mRNA and external protein. Generic simulations are carried out with 10 external scaffold proteins (i.e., 20 docking sites). https://doi.org/10.1371/journal.pcbi.1007997.g001 https://doi.org/10.1371/journal.pcbi.1007997.g001 https://doi.org/10.1371/journal.pcbi.1007997.g001 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 3 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits final complexoforms can thus consist of XX, XY, or YY. The type of genetic circuit modulates complexoform assembly Four different genetic circuits expressing proteins X and Y (parallel, cascade, series uncoupled, and series coupled) are simulated with identical first-order rates of transcription (1 t-1), translation (0.1 [C]-1t-1), export (1 t-1), and binding (1 [C]-1t-1) to 10 scaffold proteins each with 2 docking sites. The pie charts indicate the average outcomes from 1,000 stochastic simulations, showing the percentage of XX (black), YY (white), and XY/YX complexoforms (grey). The simulation results demonstrate that the genetic circuit architecture controls the final protein assembly structure. https://doi org/10 1371/journal pcbi 1007997 g002 Fig 2. Genetic circuit architecture modulates complexoform assembly. Four different genetic circuits expressing proteins X and Y (parallel, cascade, series uncoupled, and series coupled) are simulated with identical first-order rates of transcription (1 t-1), translation (0.1 [C]-1t-1), export (1 t-1), and binding (1 [C]-1t-1) to 10 scaffold proteins each with 2 docking sites. The pie charts indicate the average outcomes from 1,000 stochastic simulations, showing the percentage of XX (black), YY (white), and XY/YX complexoforms (grey). The simulation results demonstrate that the genetic circuit architecture controls the final protein assembly structure. https://doi.org/10.1371/journal.pcbi.1007997.g002 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 4 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Reaction rates of both protein expression and physical processes modulate complexoform assembly Next, we determine the impact of the reaction constants on protein assembly. We simulate each genetic circuit while individually varying the translation rate, the export rate, and the binding rate. For comparison, we also solve each condition deterministically. Fig 3 shows the mean distribution of the complexoform from stochastic simulation (circles) and deterministic solutions (lines). In all cases, the kinetic rates modulate the final distribution of the complexo- form. Slow translation, export, or binding all result in a distribution of complexoform of form. Slow translation, export, or binding all result in a distribution of complexoform of Fig 3. Computational simulations of complexoform assembly using a generic model system. Stochastic solutions (XX: black circles; YY: white circles; XY: grey circles) and deterministic solutions (XX: solid black line; YY: dashed line; XY: solid grey line) for parallel (first column), cascade (second column), series uncoupled (third column), and series coupled (fourth column). For the parallel circuit, the deterministic solutions for XX and YY overlap perfectly, so only one line is visible on the graph. For each circuit, variations in translation rate, protein export rate, and binding rate reveal an “equilibrium” regime, in which the stochastic and deterministic solutions are in relatively good agreement, and a “non-equilibrium” regime, in which the stochastic and deterministic solutions disagree. More stringent genetic circuits (e.g., the series coupled circuit) exhibits a lower discrepancy between the stochastic outcome and the deterministic solution. https://doi org/10 1371/journal pcbi 1007997 g003 Fig 3. Computational simulations of complexoform assembly using a generic model system. Stochastic solutions (XX: black circles; YY: white circles; XY: grey circles) and deterministic solutions (XX: solid black line; YY: dashed line; XY: solid grey line) for parallel (first column), cascade (second column), series uncoupled (third column), and series coupled (fourth column). For the parallel circuit, the deterministic solutions for XX and YY overlap perfectly, so only one line is visible on the graph. For each circuit, variations in translation rate, protein export rate, and binding rate reveal an “equilibrium” regime, in which the stochastic and deterministic solutions are in relatively good agreement, and a “non-equilibrium” regime, in which the stochastic and deterministic solutions disagree. More stringent genetic circuits (e.g., the series coupled circuit) exhibits a lower discrepancy between the stochastic outcome and the deterministic solution. Reaction rates of both protein expression and physical processes modulate complexoform assembly https://doi.org/10.1371/journal.pcbi.1007997.g003 5 / 18 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits roughly 25% XX, 50% XY, and 25% YY. Our results show that varying the kinetic rates in the stochastic model alters the distribution of the complexoform, demonstrating that the genetic and physical rate constants, as well as the exact genetic circuit architecture, can modulate the eventual protein assembly. We note that the rate of mRNA degradation also alters the distribu- tion of the complexoform, but that the effect is captured by altering the rate of translation (i.e., varying the ratio of the translation rate to the mRNA degradation rate), as noted in S3 Appen- dix. In contrast, the promoter binding rate, while changing the deterministic solutions, does not affect the stochastically predicted distribution of the complexoform (see S4 Appendix). As any one of the rates is increased, the stochastic distribution of the complexoform begins to deviate from the equilibrium outcome of equal amounts of X and Y in the assemblies. For the parallel circuit, any single stochastic simulation produces either predominantly XX or YY assemblies, each with a 50/50 chance, and has a very low chance of producing any heteroge- neous XY complexoforms. Since the structure of the cascade, series uncoupled, and series cou- pled genetic circuits all favor protein X expression before protein Y, higher kinetic rates tend to produce XX complexoform in these circuits. In contrast, if a sufficiently large number of both types of proteins reach the scaffold protein before the docking sites are occupied, any binding site has an equal chance of binding either protein X or Y, and the four different com- plexoforms (X+X, Y+Y, X+Y, and Y+X) have an equal chance of occurring. A homogeneous XX or YY should, therefore, each occur 25% of the time, while a heterogeneous XY or YX assembly should account for 50% of the final scaffold proteins. This 25/50/25 distribution in the final protein complex is seen in almost all cases when the rates are sufficiently slow (Fig 3). For example, a sufficiently slow rate of protein export results in the combinatorically predicted the 25/50/25 distribution for all four types of genetic circuits (Fig 3, second row). Reaction rates of both protein expression and physical processes modulate complexoform assembly In this case, the protein complex is independent of the underlying genetic circuit architecture: even though the cascade, series uncoupled, and series coupled circuits all synthesize protein X before pro- tein Y, a bottleneck in a subsequent physical process ensures that the amounts of X and Y have time to reach similar levels before accessing the docking sites. Kinetic rates drive either equilibrium or non-equilibrium assembly Our simulation results suggest that the protein assembly process can be divided into two fun- damental limiting time scales: One slow regime, in which the slow kinetic rates allow for equal amounts of protein X and Y to access the docking sites, and a fast regime, in which one protein undergoes rapid expression, transport, and binding, thus dominating the complexoform. If the assembly process is sufficiently slowed down by one or more rate-limiting steps, the pro- tein assembly process can be treated as a problem of equilibrium statistical mechanics; how- ever, this is not the case if the protein expression and assembly process are sufficiently rapid. To better quantify these two limiting time scales, which we term the “equilibrium” and “non-equilibrium” regimes, we numerically solve the set of coupled differential equations deterministically and compare our solutions to the stochastic simulation results. In the deter- ministic approach, the assembly is not limited by a small number of reactants in the system, and the deterministic numerical solutions, therefore, represent the complexoform in the limit of large-number statistical mechanics. If the stochastically driven protein assembly reaches an equilibrium state, the stochastic simulations and numerical solutions should agree. In contrast, a discrepancy between the stochastic simulations and deterministic numerical solutions would suggest that the protein assembly is not well described by predictions from large ensemble sta- tistical mechanics, but instead is governed by non-equilibrium, small-number statistics. The predictions from the deterministic solutions are shown together with the stochastic results (Fig 3). The discrepancy between the stochastic simulation average and the Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 6 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Fig 4. The discrepancy between the deterministic and stochastic solutions, and stochastic variation in protein assembly structure for four stochastically simulated genetic circuits. The discrepancy between the stochastic simulations and deterministic solutions (solid line) tends to be greater when rates are high and assembly proceeds quickly, suggesting that the protein assembly in this limit is not accurately represented by the corresponding deterministic solution. However, when assembly is slow, the predicted assembly matches the equilibrium prediction from mixing large amounts of the constituents X and Y, and all circuits yield similar protein assembly products. Circuits with a more controlled sequence of expression (e.g., the series coupled circuit) exhibit an overall better agreement with the deterministic solution. Kinetic rates drive either equilibrium or non-equilibrium assembly The stochastic variation in assembly (dashed line) also varies between the different circuits, suggesting that the type of genetic circuit, as well as the kinetics of assembly, both play into determining the variability of the assembled structure. htt //d i /10 1371/j l bi 1007997 004 Fig 4. The discrepancy between the deterministic and stochastic solutions, and stochastic variation in protein assembly structure for four stochastically simulated genetic circuits. The discrepancy between the stochastic simulations and deterministic solutions (solid line) tends to be greater when rates are high and assembly proceeds quickly, suggesting that the protein assembly in this limit is not accurately represented by the corresponding deterministic solution. However, when assembly is slow, the predicted assembly matches the equilibrium prediction from mixing large amounts of the constituents X and Y, and all circuits yield similar protein assembly products. Circuits with a more controlled sequence of expression (e.g., the series coupled circuit) exhibit an overall better agreement with the deterministic solution. The stochastic variation in assembly (dashed line) also varies between the different circuits, suggesting that the type of genetic circuit, as well as the kinetics of assembly, both play into determining the variability of the assembled structure. nistic and stochastic solutions, and stochastic variation in protein assembly structure for four stochastically simulated h l d d l ( l d l ) d b h h h d bl d Fig 4. The discrepancy between the deterministic and stochastic solutions, and stochastic variation in protein assembly structure for four stochastically simulated genetic circuits. The discrepancy between the stochastic simulations and deterministic solutions (solid line) tends to be greater when rates are high and assembly proceeds quickly, suggesting that the protein assembly in this limit is not accurately represented by the corresponding deterministic solution. However, when assembly is slow, the predicted assembly matches the equilibrium prediction from mixing large amounts of the constituents X and Y, and all circuits yield similar protein assembly products. Circuits with a more controlled sequence of expression (e.g., the series coupled circuit) exhibit an overall better agreement with the deterministic solution. The stochastic variation in assembly (dashed line) also varies between the different circuits, suggesting that the type of genetic circuit, as well as the kinetics of assembly, both play into determining the variability of the assembled structure. Kinetic rates drive either equilibrium or non-equilibrium assembly deterministic solution is calculated as the sum of the differences for each of the three types of complexoform assemblies (XX, YY, or XY/YX) (Fig 4) together with the variation in the assem- bly structure from the stochastic simulations. The stochastic simulations agree with the deter- ministic solutions in all cases and for all genetic circuits when any one of the kinetic rates is sufficiently slow, corresponding to the equilibrium regime. The equilibrium regime does not imply that all complexoform assemblies would be identical. Indeed, in the equilibrium regime, the complexoform is still stochastically determined and varies from simulation to simulation, as is evident from the non-zero stochastic variation at slow rates (Fig 4, dashed lines). How- ever, in the equilibrium regime, the average stochastic outcome is well predicted by treating the problem as a large-ensemble deterministic system. In contrast, when any one of the rates is increased, the stochastic average deviates from the deterministic solution, and the determin- istic equilibrium solution no longer accurately predicts the average complexoform from the deterministic solution is calculated as the sum of the differences for each of the three types of complexoform assemblies (XX, YY, or XY/YX) (Fig 4) together with the variation in the assem- bly structure from the stochastic simulations. The stochastic simulations agree with the deter- ministic solutions in all cases and for all genetic circuits when any one of the kinetic rates is sufficiently slow, corresponding to the equilibrium regime. The equilibrium regime does not imply that all complexoform assemblies would be identical. Indeed, in the equilibrium regime, the complexoform is still stochastically determined and varies from simulation to simulation, as is evident from the non-zero stochastic variation at slow rates (Fig 4, dashed lines). How- ever, in the equilibrium regime, the average stochastic outcome is well predicted by treating the problem as a large-ensemble deterministic system. In contrast, when any one of the rates is increased, the stochastic average deviates from the deterministic solution, and the determin- istic equilibrium solution no longer accurately predicts the average complexoform from the PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 7 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits stochastic simulation, corresponding to the non-equilibrium regime. The transition from the equilibrium to the non-equilibrium regime is seen as a rise in the discrepancy between the sto- chastic simulation average and the deterministic solution (Fig 4, solid lines). Kinetic rates drive either equilibrium or non-equilibrium assembly In the non-equi- librium regime, the protein assembly arrests into a non-equilibrium structure as vacant docking sites are depleted before reaching the equilibrium structure predicted by the deter- ministic solution. The system transitions between the equilibrium and non-equilibrium regimes when either the genetic expression rates or the physical rates of protein export and binding are altered, and generally coincides with a peak in the stochastic variation in the pro- tein assembly structure. A notable exception is observed for the simulations of the series cou- pled genetic circuit. Since this circuit imposes the most stringent structure on both the transcription and translation of the proteins, the average stochastic structure agrees well with the deterministic solution, even as the protein export or binding rate is increased. Our results also show that altering any of the rates of the system, in turn, changes the aver- age total time to fully occupy the available docking sites, which we term time to assembly (Tasb). The average fraction of XX, XY, and YY complexoform in the stochastic simulation is shown as a function of the average Tasb in Fig 5. Contrary to our expectation, the fraction of each complexoform falls onto a single curve, regardless of which of the rates is being changed. Although creating an earlier bottleneck by slowing down the earlier processes, such as transla- tion, results in the longest Tasb, the total Tasb is highly correlated with the complexoform com- position, and the average assembly outcome from the stochastic simulation can be correlated to a single parameter, the Tasb. Here again, however, the series coupled genetic circuit is an exception; when the translation rate in this genetic circuit is lowered, the sequential architec- ture of the circuit results predominantly in XX complexoform. This result again suggests that the gene circuit architecture can exert a high level of control over the protein assembly struc- ture, even when the assembly is slow, and a large number of proteins are involved in stochasti- cally forming the structure. Comparisons to other biological model systems To compare our modeling results to typical rates and concentrations in biological systems, and to determine whether protein assembly in biological systems generally fall in the equilibrium or non-equilibrium regime, we run our computational models with typical biochemical and physical rates of protein expression, export, and binding for bacterial cellulosome. This system is modeled as a scaffold protein with 10 binding sites to which proteins X and Y can bind. Rates representative of the bacterial cellulosome complexoform, as well as relevant references, are summarized in Table 1. Since it is difficult to estimate the exact external per-molecule binding rate and loss rate of individual proteins, we simulate cellulosome assembly with a range of possible protein binding rates while keeping the loss rate fixed, thus varying the ratio of binding to loss rate for the external protein. Each condition is simulated 1,000 times to gen- erate a histogram of the outcomes, as quantified by the number of binding sites on the scaffold protein occupied by protein X (the rest being occupied by protein Y; see Fig 6). From these simulations, we quantify both the variation in stochastic assembly, as well as the discrepancy between stochastic simulations and deterministic solutions. The simulation results for each of the four genetic circuits (parallel, cascade, series uncoupled, and series coupled) are summa- rized in Fig 7, with each of the translation rate, protein export rate, and protein binding rate being varied about the estimated physiological value from literature and previous experimen- tally reported values. Our simulations show that, as observed for the generic model system, the type of genetic circuit exerts a great deal of control over the eventual protein assembly. For example, a parallel Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 8 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits circuit will result in an average assembly of 50% X and 50% Y (Fig 7, column 1), whereas the Fig 5. The genetic circuits generate similar assembly times, regardless of which rate is varied. Shown is the stochastically simulated average fraction of XX (black), YY (white), and XY complexoform (grey) in the protein assemblies when varying translation rate (circles), protein export rate (squares), and protein binding rates (diamonds). Each circuit exhibits a transition from the fast non-equilibrium regime to the slow equilibrium regime, although the exact transition time is dependent on the type of genetic circuit. Comparisons to other biological model systems A notable exception is the series coupled circuit, indicating that the highly sequential architecture of this circuit modulates the protein assembly structure in both the fast and slow regimes. https://doi.org/10.1371/journal.pcbi.1007997.g005 Fig 5. The genetic circuits generate similar assembly times, regardless of which rate is varied. Shown is the stochastically simulated average fraction of XX (black), YY (white), and XY complexoform (grey) in the protein assemblies when varying translation rate (circles), protein export rate (squares), and protein binding rates (diamonds). Each circuit exhibits a transition from the fast non-equilibrium regime to the slow equilibrium regime, although the exact transition time is dependent on the type of genetic circuit. A notable exception is the series coupled circuit, indicating that the highly sequential architecture of this circuit modulates the protein assembly structure in both the fast and slow regimes. https://doi.org/10.1371/journal.pcbi.1007997.g005 https://doi.org/10.1371/journal.pcbi.1007997.g005 circuit will result in an average assembly of 50% X and 50% Y (Fig 7, column 1), whereas the cascade circuit and the series circuits are weighted towards X to varying degrees (Fig 7, col- umns 2, 3, and 4). As was also observed for the generic model, the parallel genetic circuit exhibits the highest degree of variation in the assembly outcome, whereas the more structured genetic circuits have lower variability in the final assembly composition. One major difference between the generic model (with arbitrary rates) and the cellulosome model is that whereas the stochastic simulations of the generic model would often deviate significantly from the deterministic solutions, the simulations with the rates for the cellulosome system exhibits vir- tually no such deviations between the stochastic average and the deterministic solution. One exception to this is the parallel circuit, where a very fast rate of expression could have resulted in an assembly consisting of either X or Y (see also Fig 6C). Overall, however, the model using biological cellulosome rates predicts good prediction between the deterministic and stochastic circuit will result in an average assembly of 50% X and 50% Y (Fig 7, column 1), whereas the cascade circuit and the series circuits are weighted towards X to varying degrees (Fig 7, col- umns 2, 3, and 4). As was also observed for the generic model, the parallel genetic circuit exhibits the highest degree of variation in the assembly outcome, whereas the more structured genetic circuits have lower variability in the final assembly composition. Comparisons to other biological model systems One major difference between the generic model (with arbitrary rates) and the cellulosome model is that whereas the stochastic simulations of the generic model would often deviate significantly from the deterministic solutions, the simulations with the rates for the cellulosome system exhibits vir- tually no such deviations between the stochastic average and the deterministic solution. One exception to this is the parallel circuit, where a very fast rate of expression could have resulted in an assembly consisting of either X or Y (see also Fig 6C). Overall, however, the model using biological cellulosome rates predicts good prediction between the deterministic and stochastic PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 9 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Table 1. Rates and concentrations used in simulating cellulosome assembly. Value References Rates kpro (RNAP promoter binding) 5.6107 M-1s-1 = 0.09 s-1 [16] kpro- (RNAP promoter unbinding) 0.20 s-1 kgene (RNAP synthesis rate constant) 0.36 s-1 [17] km (transcription) 0.03 s-1 [17–20] kmrna-loss (mRNA degradation) 0.002 s-1 [21] kp (translation) 0.03 s-1 [19,20,22] kout (diffusive protein export) 0.5 s-1 [23–24] kbind (protein association rate) 1106 M-1s-1 = 0.002 s-1 [25–26] kbind- (protein dissociation rate) 0 kout-loss (protein loss) 0.0002 s-1 [27] Reactants P (promoter DNA) 1 RNAP (RNA polymerase) 100 [28] Rib (ribosome) 5 [29–30] Docking sites per scaffold protein 10 [5] Note: Two proteins are expressed and bind to an external scaffold with 10 binding sites. Intracellular molar t ti t d t l l ll i th t 1 M i l t 6 108 l l ll N t th t Table 1. Rates and concentrations used in simulating cellulosome assembly. Note: Two proteins are expressed and bind to an external scaffold with 10 binding sites. Intracellular molar concentrations are converted to molecules per cell assuming that 1 M is equal to 6108 molecules per cell. Note that the protein assembly process is assumed to be irreversible for the purposes of this simulation, whereas actual measurements in the literature does show that the complex does disassemble over time. https://doi.org/10.1371/journal.pcbi.1007997.t001 simulations, although the stochastic simulations still predict significant variability in the final protein assembly. Interestingly, the simulations also reveal that the cellulosome kinetic rates place the system at the transition between the fast and slow limits seen in the generic model. Comparisons to other biological model systems For example, in the cascade genetic circuit, the biological rates predict an average assembly simulations, although the stochastic simulations still predict significant variability in the final protein assembly. Interestingly, the simulations also reveal that the cellulosome kinetic rates place the system at the transition between the fast and slow limits seen in the generic model. For example, in the cascade genetic circuit, the biological rates predict an average assembly Fig 6. Stochastic simulations and deterministic solutions for a cellulosome model system, in which two proteins X and Y expressed in a parallel gene circuit bind to 10 scaffold binding sites to form a complexoform. The outcome of 1,000 stochastic simulations is indicated as grey bars, while the deterministic solution is indicated by a dashed line. The model parameters are summarized in Table 1, with the translation rate constant in this figure being varied across several orders of magnitude: 3.10-5 s-1 (A), 3.10-2 s-1 (B), and 3.101 s-1 (C). For each simulation condition, the stochastic variation in cellulosome assembly is calculated as the standard deviation of the distribution of stochastic simulations. The discrepancy between the stochastic simulations and deterministic solution is calculated as the average difference between each simulation and the deterministic solution. https://doi org/10 1371/journal pcbi 1007997 g006 Fig 6. Stochastic simulations and deterministic solutions for a cellulosome model system, in which two proteins X and Y expressed in a parallel gene circuit bind to 10 scaffold binding sites to form a complexoform. The outcome of 1,000 stochastic simulations is indicated as grey bars, while the deterministic solution is indicated by a dashed line. The model parameters are summarized in Table 1, with the translation rate constant in this figure being varied across several orders of magnitude: 3.10-5 s-1 (A), 3.10-2 s-1 (B), and 3.101 s-1 (C). For each simulation condition, the stochastic variation in cellulosome assembly is calculated as the standard deviation of the distribution of stochastic simulations. The discrepancy between the stochastic simulations and deterministic solution is calculated as the average difference between each simulation and the deterministic solution. https://doi.org/10.1371/journal.pcbi.1007997.g006 https://doi.org/10.1371/journal.pcbi.1007997.g006 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 10 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Fig 7. Computational simulations of cellulosome assembly consisting of a scaffold with 10 available binding sites for proteins X and Y for four stochastically simulated genetic circuits. The model parameters are summarized in Table 1. Comparisons to other biological model systems The vertical axis indicates the number of scaffold binding sites occupied by protein X. The average of 1,000 stochastic simulations is indicated by solid circles for each condition, with error bars indicating the standard deviation from the simulation. Deterministic solution results are indicated by dashed lines. The discrepancy between stochastic and deterministic solutions (calculated as the average difference between each simulation and the deterministic solution) is indicated by solid lines. Grey dots indicate the estimate of physiological rates (Table 1), about which each rate is varied. Fig 7. Computational simulations of cellulosome assembly consisting of a scaffold with 10 available binding sites for proteins X and Y for four stochastically simulated genetic circuits. The model parameters are summarized in Table 1. The vertical axis indicates the number of scaffold binding sites occupied by protein X. The average of 1,000 stochastic simulations is indicated by solid circles for each condition, with error bars indicating the standard deviation from the simulation. Deterministic solution results are indicated by dashed lines. The discrepancy between stochastic and deterministic solutions (calculated as the average difference between each simulation and the deterministic solution) is indicated by solid lines. Grey dots indicate the estimate of physiological rates (Table 1), about which each rate is varied. https://doi.org/10.1371/journal.pcbi.1007997.g007 with about 90% protein X; however, if the binding rate had been 10-fold slower (or the loss rate 10-fold higher), the fraction of protein X in the assembly drops to 77%, and a 1000-fold change predicts a fraction of 56% X, i.e., almost equal amounts of X and Y. Both protein bind- ing and protein loss could depend on the exact extracellular conditions (e.g., the degree of crowding), thus impacting the assembly purely by altering the kinetic rates involved. Thus, our data demonstrate that both the kinetic rates and the genetic circuit can moderate the eventual assembly outcome in the case of the bacterial cellulosome. Discussion In summary, our work shows that the underlying genetic circuit architecture does modulate the protein assembly. However, it is the interplay between the circuit architecture and the genetic and physical rate kinetics that together determine the protein assembly structure. We demonstrate two distinct behaviors of kinetic assembly: a slow equilibrium regime, in which the average assembly is well described by equilibrium statistical mechanics, and a fast non- equilibrium regime, in which the average assembly arrests before the system reaches equilib- rium. Regardless of the equilibrium or non-equilibrium regime, the cumulative protein PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 11 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits concentrations (i.e., the total amount of protein available to bind over time) determine the eventual complexoform distribution (S5 Appendix). Furthermore, we demonstrate that the two regimes can be regulated by tuning any of the kinetic rates involved in the protein expres- sion and assembly process, whether biochemical or physical. The arresting of the assembly into a non-equilibrium structure has previously been observed on much larger length scales, such as in the dynamic arrest occurring in macroscopic protein assemblies such as biopolymer networks, in which the kinetics of assembly can highly affect the non-equilibrium assembly structure [31–34]. Our work shows that similar dynamic arrest can occur on a much smaller scale as well, in the assembly of protein complexes involving just a handful of individual pro- teins. The results highlight new mechanisms, in addition to restrictive or preferential binding, through which systems can control stochastic processes such as protein assembly. The results also underscore the importance of both kinetics and stochastic non-equilibrium behavior in addition to the genetic circuit architecture as modulators of protein assembly processes. There are several future considerations for our results. First, our work did not consider the details of how genetic context affects the transcription or translation rate of genes. For instance, a prior work [35] shows that the arrangement of two genes can affect the level, dynamic range, and sensitivity of their expression. The spatial arrangement of the two genes will affect the expression of our parallel genetic module. Specifically, if the parallel genetic module is inducible by certain chemicals, its sensitivity and dynamic range of induction would also be affected by the arrangement of the two genes, as suggested by the prior work. Discussion In our work, we have not considered the induction dynamics of the genetic module, which may serve as another regulatory force of complexoforms. Second, one challenge in our work is to obtain the analytical solution of the stochastic equa- tions. For instance, previous work by Laurenzi, Renyi, Mcquarrie, and Ishida [36–39] have solved the analytical solutions for A+B$C. To obtain the solution, the authors exploit the con- servation of mass between A, B, and C. Finding similar analytical solutions to our model involves two challenges. First, there are multiple bimolecular reactions in our model. Second, it is challenging to incorporate the conservation of mass for intermediate reactions of our model. There is, however, a qualitative agreement between our simulations and prior analytical solution. For instance, Laurenzi shows that stochastic solution deviates from a deterministic solution when the number of molecules is low. We observe this trend generally for all our results because the molecular number in our system is low. Furthermore, Laurenzi shows that the variance of molecular concentrations peaks at intermediate reaction time when the reac- tion becomes irreversible. Likewise, we observe that when the reaction rate becomes slower, our system takes a long time to approach the steady-state solution. The longer transient kinet- ics increase the duration when the system exhibits a high variance of molecular concentration. This enhanced variance likely leads to the significant deviation between the stochastic and deterministic solutions when reaction rate constants are high. Third, the arrangement of two genes in the series coupled module may affect the expression level of the genes. Typically, the second gene is expressed at a lower level than the first gene. A prior work [40] has quantified the effect of GC content, size, and folding energy of the first gene on the expression of the second gene. Incorporating these details will improve the predic- tion power of our model on the distribution of complexoforms. Our study points to the needs of quantifying heterogeneity of complexoform inside cells and studying the regulatory mechanisms of the heterogeneity. New mass spectrometry tech- nology and modeling tools may be used to reveal new insights into complexoforms that were not possible before. For instance, the proteasomes of Rhodococcus consisted of both alpha (α1, α2) and beta (β1, β2) subunits [41]. The subunits can form four variants of complexoform (α1β1, α1β2, α1β2, α2β1) with similar proteasome functions. Discussion Does natural Rhodococcus control PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 12 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits the heterogeneity of the proteasome at the genetic or protein level, or both? The Trp synthase of E. Coli also consists of two α and two β subunits. The genes are arranged in the same order as the assembly order of the subunits [42]. Does the assembly process of the synthase fall in the equilibrium or non-equilibrium regimes? Answer to the question requires detailed measure- ments of the kinetic parameters that are not available yet. Finally, the Pili of E. Coli consists of four different protein subunits, and the gene order is arranged opposite of the assembly order [43]. Does this contradiction of gene and assembly order indicate that the assembly is solely determined by protein-protein binding and that there is less heterogeneity in the final protein complex? To this end, when the kinetic parameters of these processes become available, our modeling framework may be used to predict the heterogeneity of the complexoform and the equilibrium vs. non-equilibrium control of the systems. Furthermore, a meta-analysis of genome sequences and protein-protein interaction maps may be used to reveal the relative abundance of each circuit architectures for assembling known complexoforms. Modeling protein expression, export, and binding Our computational model consists of a set of coupled biochemical and physical processes, starting from transcription, and ending with the binding of protein products to 10 scaffold proteins, each with two docking sites to form a final complexoform. In the first process, an RNA polymerase binds to a promoter to synthesize mRNA. This transcription step is followed by the translation of two different protein products, denoted X and Y. The proteins are then transported for binding to a scaffold protein to form a three-protein complexoform. The model, which is summarized schematically in Fig 1, also incorporates the degradation of mRNA and diffusive loss of proteins after transport. The transcription and translation of the two proteins occur through one of four genetic cir- cuits: (1) a parallel expression circuit, in which transcription and translation of the two pro- teins happen concurrently and independently; (2) a cascade expression circuit, in which the transcription and translation of protein X create the needed transcription activator for the sub- sequent transcription of protein Y; (3) a series uncoupled expression circuit, in which the tran- scription of the two genes occurs sequentially but with uncoupled translation; and (4) a series coupled expression circuit, in which both the transcription and translation are sequential. Within each of these four different genetic circuit architectures, the rates of genetic expression and physical export and protein binding processes are varied in the model. In all cases, the evo- lution of the system is arrested as the docking sites are exhausted, and no further changes to the protein assembly structure can occur once they are all occupied. The set of biochemical and physical processes defining each genetic circuit is listed in S1 Appendix. All processes are modeled as linear reactions, in which the reaction is linearly dependent on a rate constant and on the concentration of the reactants. Thus, for a set of reac- tions of the form A þ B þ ! k1 Q Q þ R þ ! k2 Z A þ B þ ! k1 Q Q þ R þ ! PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 Computational model implementation All computer simulations are done using Matlab (Mathworks, Natick MA). The built-in ode45 differential equation solver is used for all deterministic numerical solutions. The initial condi- tions of the deterministic differential equation solutions are identical to those of the stochastic model, except that the amount of each reactant is treated as a continuous, rather than a discrete variable, and are allowed non-integer values. Stochastic simulations use a custom-written Gillespie algorithm [44] in Matlab, in which each reaction outlined in S1 Appendix is treated stochastically. In this algorithm, each possible reaction is numerically simulated using a Monte Carlo technique by generating a tentative reaction time for each reaction based on the concentrations and reaction rates at that instant in time, and in each case, executing the reaction with the shortest tentative reaction time. This process is repeated until all binding sites are occupied, after which no further evolution of the protein assembly can occur. Unlike in the deterministic solution, the stochastic simulation only allows for discrete changes in the amount of each reactant, and only integer values are allowed for each reactant. To account for the stochastic variation between simulations, each set of conditions is simulated 1,000 times and averaged. For each set of initial conditions, we calculate the average assembly structure for the stochastic simulation by counting the docking site occupancy (i.e., the final average number of scaffold proteins with XX, XY, or YY count for each run of the simulation). The standard deviation is calculated for each of the three types of assemblies. We quantify the stochastic variation in the simulation as the sum of these three standard deviations. To compare the deterministic and stochastic solutions, we calculate the difference between the amounts of each type of protein complex predicted from the deterministic solution with the average amount observed from the stochastic simulation. The sum of these three differ- ences is used as a measure of the overall discrepancy between the deterministic and stochastic computational solutions. For each genetic circuit, we vary the translation rate, the protein export rate, and the protein binding rate to investigate the impact of kinetics on the modulation of the final protein assem- bly. The initial conditions and rates are all listed in S2 Appendix. Modeling protein expression, export, and binding k2 Z the rate of change of any reactant is described by a first-order differential equation: d½Q dt ¼ þk1 A ½ B ½ k2 Q ½ R ½ 13 / 18 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits Here, [] indicates the concentration of each reactant. Any additional reactions involving the reactant Q is incorporated as additional, linear terms, and the rate of change of each reactant is modeled by an additional differential equation. In this model, any arbitrary number of reac- tions involving any number of reactants can thus be written as a set of coupled, first-order dif- ferential equations. For large ensembles (i.e., large concentrations of all reactants involved), these coupled differential equations are deterministic, and the outcome for each reactant can be predicted exactly by solving the equations numerically. However, for a sufficiently small number of molecules, the outcome becomes stochastic and must be approached using stochas- tic algorithms. We, therefore, solve the coupled differential equations both deterministically to obtain exact numerical solutions, and stochastically using a stochastic algorithm. Computational model implementation In order to investigate the impact of the relative, rather than the absolute, kinetic rates on the final assembly structure in the model system, simulations are done using arbitrary but dimensional units: time is indi- cated in units of t, concentrations are indicated in units of [C], and rates are indicated in units of [C]-1t-1 or t-1. Biological rates of the cellulosome complex Alth h th t ll l l t i t f t l ti d l d f Although the exact cellulosome complex can contain many types of catalytic modules and scaf- foldin binding sites [5], we adopt a model in which the external scaffold has 10 available sites to which two different proteins can directly bind, as this model captures the competing role of both timing and gene circuit architecture in modulating the complexoform. While some rates relevant to complexoform enzyme expression and assembly have been previously measured and reported in the literature, such as the promoter binding and unbinding rates [16] and the rates of the protein-protein binding interaction [25–26], we infer or estimate other rates to an order of magnitude, as described below. Previous reports have shown that mRNA degradation occurs with half-lives ranging from a few minutes in E. Coli [21] and ~20 minutes in S. Cerevisiae yeast [45] to several hours in H. Sapiens cells [46]. Therefore, we assume a typical bacterial mRNA half-life of 5 minutes, corre- sponding to a decay rate of 0.002 s−1. The rate of protein export is assumed for on the size of 50–100 kDa [23], suggesting an effective diffusion coefficient of about 1 μm2s-1 through a cytoplasmic environment [24]. We, therefore, estimate that diffusive transport in a bacterium with a size of ~2 μm would take on the order of 2 seconds, corresponding to a rate of export on the order of 0.5 s-1 after completed expression. We assume that RNA polymerase creates RNA at a rate of about 45 nucleotides per second, and that ribosomal translation occurs at a rate of about 15 amino acids per second [17–20,22]. Since 3 base pairs code for one amino acid, the rate of transcription and translation are roughly equal. Assuming a typical protein size of about 50 kDa and an average amino acid size of 110 Da, one full protein would take 30 seconds of transcription and translation, corresponding to a rate of 0.03 s-1 for both processes. In order to consider reaction rates and concentrations in terms of the number of molecules of each reactant inside a bacterium, we convert molar concentrations to molecules per bacteria volume with E. Coli as a reference. A concentration of 1 M corresponds to 6108 molecules/ bacterium, assuming a bacterial volume of 1 fL. PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 Biological rates of the cellulosome complex In order to compare our simulation results in arbitrary units to biological systems, we run both deterministic and stochastic simulations using typical concentrations and biochemical 14 / 18 PLOS Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits and physical rates of protein expression, export, and binding for the bacterial cellulose-degrad- ing multi-enzyme complexes, known as cellulosomes. We modeled the assembly of cellulo- somes, which comprise scaffold proteins containing repeated copies of Cohesin (Coh) domains and catalytic enzymes containing Dockerin (Doc) domains. Each Coh within the scaffold serves as a binding site for the corresponding Doc domain located at the C-terminus of each enzyme. The enzymes are secreted from the bacteria and assemble onto the extracellu- lar scaffold proteins through Coh-Doc binding. The Coh-Doc interactions, albeit high-affinity, are promiscuous, and dozens of different enzymes containing homologous and closely related terminal Doc sequences are able to bind to any of the Coh domains within the scaffold. Although the exact cellulosome complex can contain many types of catalytic modules and scaf- foldin binding sites [5], we adopt a model in which the external scaffold has 10 available sites to which two different proteins can directly bind, as this model captures the competing role of both timing and gene circuit architecture in modulating the complexoform. While some rates relevant to complexoform enzyme expression and assembly have been previously measured and reported in the literature, such as the promoter binding and unbinding rates [16] and the rates of the protein-protein binding interaction [25–26], we infer or estimate other rates to an order of magnitude, as described below. and physical rates of protein expression, export, and binding for the bacterial cellulose-degrad- ing multi-enzyme complexes, known as cellulosomes. We modeled the assembly of cellulo- somes, which comprise scaffold proteins containing repeated copies of Cohesin (Coh) domains and catalytic enzymes containing Dockerin (Doc) domains. Each Coh within the scaffold serves as a binding site for the corresponding Doc domain located at the C-terminus of each enzyme. The enzymes are secreted from the bacteria and assemble onto the extracellu- lar scaffold proteins through Coh-Doc binding. The Coh-Doc interactions, albeit high-affinity, are promiscuous, and dozens of different enzymes containing homologous and closely related terminal Doc sequences are able to bind to any of the Coh domains within the scaffold. Author Contributions Conceptualization: Michael A. Nash, Cheemeng Tan. Formal analysis: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Funding acquisition: Michael A. Nash, Cheemeng Tan. Investigation: Mikkel Herholdt Jensen, Eliza J. Morris, Hai Tran. Methodology: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Software: Mikkel Herholdt Jensen, Eliza J. Morris, Hai Tran, Cheemeng Tan. Validation: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Writing – original draft: Mikkel Herholdt Jensen, Cheemeng Tan. Writing – review & editing: Mikkel Herholdt Jensen, Eliza J. Morris, Hai Tran, Micha Nash, Cheemeng Tan. Conceptualization: Michael A. Nash, Cheemeng Tan. Formal analysis: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Methodology: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Methodology: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Software: Mikkel Herholdt Jensen, Eliza J. Morris, Hai Tran, Cheemeng Tan. Validation: Mikkel Herholdt Jensen, Eliza J. Morris, Cheemeng Tan. Writing – original draft: Mikkel Herholdt Jensen, Cheemeng Tan. Writing – review & editing: Mikkel Herholdt Jensen, Eliza J. Morris, Hai Tran, Michael A. Nash, Cheemeng Tan. Biological rates of the cellulosome complex We assume one copy of each gene (typically the case for bacteria with one chromosome), with about 100 RNA polymerases per gene [28] (estimated from ~23 RNA polymerase per lacZ gene). Although there are about 20,000 ribo- somes in one E. Coli bacterium [29], these are employed in the expression of roughly 4,000 dif- ferent genes [30]; we, therefore, assume 5 available ribosomes. It is difficult to estimate the exact external per-molecule binding rate and loss rate of indi- vidual proteins. The cohesin-dockerin association rate has been measured to be about 1106 M-1s-1 [25–26], and assuming that the outside reaction volume is similar to the volume of a single bacterium, the external binding rate is 0.002 s-1 per molecule. Protein lifetimes are typi- cally on the order of tens of minutes or several hours [27]. Assuming a lifetime of about an hour, this implies a protein loss rate of 0.0002 s-1 per molecule. All the values used for the cellulosome assembly simulations are summarized in Table 1. Since most values are estimated to an order of magnitude, we vary the translation, export, and binding rates across many orders of magnitude to elucidate whether the cellulosome Computational Biology \| https://doi.org/10.1371/journal.pcbi.1007997 June 29, 2020 15 / 18 PLOS COMPUTATIONAL BIOLOGY Stochastic ordering of protein assemblies by genetic circuits complexoform assembly process occurs in the equilibrium or non-equilibrium regime. In the simulation, we vary the ratio of the protein association rate and diffusive loss rate. Although data shows a slow dissociation of the dockerin/cohesin complex, we assume that the binding is irreversible to allow us to study the structure of the completed assembly right after protein expression, transport, and assembly. We note that with a slowly reversible binding, the com- plexoform distribution should eventually equilibrate with the external solution concentrations of protein in solution over long time scales. All data generated for and presented in this study are available in full in the S1 Data. 2. Skinner OS, Havugimana PC, Haverland NA, Fornelli L, Early BP, Greer JB, et al. An informatic frame- work for decoding protein complexes by top-down mass spectrometry. Nature methods. 2016; 13 (3):237. https://doi.org/10.1038/nmeth.3731 PMID: 26780093 Supporting information S1 Appendix. (PDF) S2 Appendix. (PDF) S3 Appendix. (PDF) S4 Appendix. (PDF) S5 Appendix. (PDF) S1 Data. 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hal-00763812-2012_ISCAS_Palumbo.txt_1	French-Science-Pile	Open Science	Various open science	2,012	Multi-Purpose Systems: A Novel Dataflow-Based Generation and Mapping Strategy. 2012 IEEE International Symposium of Circuits and Systems, May 2012, South Korea. pp.NC. ⟨hal-00763812⟩	None	English	Spoken	2,619	4,808	Multi-Purpose Systems: A Novel Dataflow-Based Generation and Mapping Strategy Nicolas Siret, Matthieu Wipliez, Jean François Nezan, Francesca Palumbo, Luigi Raffo To cite this version: Nicolas Siret, Matthieu Wipliez, Jean François Nezan, Francesca Palumbo, Luigi Raffo. Multi-Purpose Systems: A Novel Dataflow-Based Generation and Mapping Strategy. 2012 IEEE International Symposium of Circuits and Systems, May 2012, South Korea. pp.NC. �hal-00763812� HAL Id: hal-00763812 https://hal.science/hal-00763812 Submitted on 11 Dec 2012 HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Multi-Purpose Systems: A Novel Dataflow-Based Generation and Mapping Strategy Nicolas Siret, Matthieu Wipliez and Jean-François Nezan Francesca Palumbo and Luigi Raffo European university of Brittany, France INSA, IETR, UMR 6164, F-35708 RENNES Email: name.surname@insa-rennes.fr DIEE - University of Cagliari, Italy Piazza D’Armi 09123 Cagliari Email: <francesca.palumbo; luigi>@diee.unica.it Abstract— The Dataflow Process Networks (DPN) Model of Computation (MoC) has been used in different ways to improve time-to-market for complex multipurpose systems. The development of such systems presents mainly two problems: (1) the manual creation of the multi-purpose specialized hardware infrastructures is quite error-prone and may take a lot of time for debugging; (2) the more hardware are the details to be handled the greater the effort required to define an optimized components library. This paper tackles both problems, leveraging on the combination of the DPN MoC with a coarse-grained reconfigurable approach to hardware design and on the exploitation of the DPN MoC for the synthesis of target-independent hardware codes. Combining two state of the art tools, namely the Multi-Dataflow Composer tool and the Open RVCCAL Compiler, we propose a novel dataflow-based design flow that provide a considerable on-chip area saving targeting both FPGAs and ASICs. I. Introduction. The embedded systems world in the last few years has changed: new systems are required to be more flexible with respect to their predecessors. Standards in video codecs and telecommunication infrastructures are constantly evolving and applications are getting more complex every day. These requirements led to the development of complex, heterogeneous, multi-purpose systems. An optimal solution to target them is offered by the reconfigurable hardware design paradigm, able to guarantee: (1) flexibility (2) high performance (through native specialization capabilities) and (3) possible long-term adaptability. One of the most critical aspect with such highly specialized platforms turned out to be the compilation toolchain: dealing with multi-purpose systems, the definition of the proper bit-streams set for platform configuration is not trivial [1], [2]. Moreover, the more is the specialization the more can be the time required to develop and debug the complex and heterogeneous Funtional Units (FUs) that may compose the overall platform. In this paper we aim at addressing the abovementioned issues to decrease the time-to-market of modern multi-purpose computational intensive systems, leveraging mainly on the Dataflow Process Networks (DPN) Model of Computation (MoC). The main novelty of this research resides in proposing a novel design flow for multi-purpose reconfigurable systems, exploiting two state of the art tools: the Multi-Dataflow Composer, MDC tool, and the Open RVC-CAL Compiler, Orcc. Coarse-grained runtime reconfiguration (CGRR) and the DPN MoC have been combined to address the tricky problem of assembling, in a reasonable time and without any user intervention, specialized and flexible CGRR platforms automatically mapping on them complex software applications. The rest of this paper is organized as follow: Sect. II provides an overview of the MDC tool and Orcc, together with the description of the proposed dataflow-based design paradigm, Sect. III discusses the achieved performance and Sect. IV concludes with some final remarks. II. Dataflow-Based Design Flow The main goal of the proposed novel design flow is the reduction of the time-to-market for the design of complex multi-purpose reconfigurable systems, combining together two approaches based on the DPN MoC: the MDC and Orcc. A. The Multi-Dataflow Composer tool The Multi-Dataflow Composer tool exploits the composability property of the DPN models to enable high level dataflow descriptions to be mapped directly on hardware. Originally conceived to be compliant to MPEG Reconfigurable Video Coding (RVC) [3] applications, it was then extended to any RVC compliant application [4], [5]. The MDC analyses input dataflows, recognizes similarities among them and assembles a unique HDL datapath where the common FUs are shared, through the intervention of dedicated data switching units (the Sbox units in Fig.1b). The output of the MDC is the multipurpose CGRR platform implementing all the given input networks. The Sbox units allow on-the-fly switching among the implemented dataflows without any low-level reconfiguration. This reconfiguration mechanism, provides an efficient and quick substrate reconfiguration and allows the multi-purpose system to switch in a single clock cycle among the dataflows, without any shut-down overhead. Moreover, the produced CGRR platform is completely target-independent. D A A F G A C B E Dataflow network 2 Dataflow network 3 E G B E Orcc-VHDL B Dataflow network 1 N:N Orcc-VHDL N:N STEP-1 Dataflow network 3 Dataflow network 1 RVC– CAL compliant files G Dataflow network 2 C B F D A E G Actors: .vhd files STEP-1 MDC tool A A D F N:1 G Sbox Units: .v files Multi-Purpose CGRR Platform C HDL code 2 B LUT E Configuration B E G HDL code 1 HDL code 3 HDL Synthesis A Sbox B Sbox Sbox E C D F STEP-2 Sbox G Post-optimizations Orcc-VHDL STEP-2 Hardware ASIC-FPGA (a) Orcc-VHDL Synthesis Flow. Fig. 1. (b) Dataflow-Based Novel Design Flow. Summary of Design Flows. B. The Open RVC-CAL Compiler Orcc was initially conceived to produce software code from dataflow programs in the RVC domain [6], [7]. It has been recently extended, to implement an Electronics System Level Design (ESLD) framework, by adding the High Level Synthesis (HLS). The ESLD is a recent methodology which aims at improving the productivity of system architects by offering an efficient support of hardware/software co-designing, and providing HLS [8]. In one word, HLS tools compile high-level of abstraction programs into synthesizable hardware codes. Thereby, Orcc (namely Orcc-VHDL for the hardware code generation) generates a target-independent hardware code [9] which can be simulated and synthesized on any kind of hardware platforms. In our approach of the HLS, a dataflow program is compiled in hardware code in two-steps, depicted in Fig 1a. First the VHDL code is generated using Orcc-VHDL, then it is synthesized for a specific target using any state of the art HDL-to-gates synthesizers. The main advantages of such two-steps approach are: (1) the generated VHDL is understandable, updatable and portable, and, (2) the design flow is closer to the one designers are accustomed to. C. Novel Design Flow Fig.1b presents the novel dataflow-based design toolchain proposed in this paper. By coupling the MDC tool and Orcc-VHDL to process a common set of RVCCAL compliant networks, it is possible (without users intervention) to: (1) automatically derive the HDL CGRR platform of the multi-purpose system (MDC), (2) to generate the hardware library of FUs (Orcc-VHDL) and (3) to handle the on-the-fly runtime programmability (MDC). Since both the MDC tool and Orcc-VHDL generate target-independent hardware code, the provided RTL description is compliant with any (recent) synthesizing tools. Therefore, runtime reconfiguration on the given coarsegrained substrate can take place both on ASICs and on FPGAs with the following benefits: ASIC CGRR provides high performance and gains in flexibility. • FPGA CGRR allows resource sharing, so that a smaller FPGA may be used, while reducing power consumption and avoiding the overhead of a partial/complete context switch. • The availability of a complete composition and generation framework for multi-purpose heterogeneous systems based on a common formalism, constituted by the DPN MoC and the RVC-CAL standard, enables to move some steps towards the possibility of closing the gap between hardware and software, natively addressing the tricky problem of automatically mapping software on highly specialized heterogeneous hardware. Moreover, this approach has to be considered orthogonal to different reference domains. In fact, by simply providing the applications according to the reference formalism, any multi-flow system can be synthesized. D. Comparison with Related Works Other automated tools have been developed to automatically map applications on a coarse-grained reconfigurable system. Nevertheless, as far as we know none of them is able to automatically derive the library of FUs as long as This section presents the results obtained adopting the proposed dataflow-based design flow on two different multi-standard platforms: the first one running a very simple 1-D Inverse Discrete Cosine Transform (IDCT1D), while the second a 2-D Inverse Discrete Cosine Transform (IDCT2D). The IDCT1D is a static dataflow network and a reference application in signal and image processing. The IDCT2D isa hierarchical dynamic dataflow network that executes two IDCT1D and transposes the signal between the two executions. A. Performance on the IDCT1D On the top of Fig. 2 two different IDCT1D dataflow models are presented, whereas on the bottom their combination. The original IDCT1D dataflow networks share just one single actor; therefore, the benefits of assembling a CGRR platform are not straightforward. However, this use case is sufficient to show the validity the approach with respect to the one cycle runtime reconfiguration. Moreover, despite the number of inserted Sbox units (7) is bigger than the number of FUs (1) placed in common, both on a Xilinx Virtex6 and an Altera Spartan3 FPGAs it was possible to save around the 5% of area. B. Performance on the IDCT2D To implement a multi-standard version of the IDCT2D we have decided to assemble a CGRR architecture (CGRRA) able use alternatively both the IDCT1D models presented on the top of Fig. 2. This platform, for exploration purposes, has been compared to a static dual mode architecture (SDMA). Table I summarizes the overall number of involved FUs, as long as the overheads required for the SDMA and for the CGRRA. Table II summarizes the performance obtained using the Altera Quartus II and the Xilinx ISE synthesisers for both the CGRRA and the SDMA. The increased complexity due to Sbox units insertion, when FUs are shared, leads to a very small penalty in terms of operating frequency, while area benefits are very promising: nearly 31% fewer FUs Sboxes LUTs IDCT 2D model 1 IDCT 2D model 2 SMDA CGRA 11 12 23 15 0 0 0 7 0 0 0 1 TABLE I slices and, respectively, 28% (Quartus) and 23% (ISE) fewer LUTs. CGRA Benefit Max. Frequency [MHz] Slices [#] LUTs [#] Power Dissip. [mW] 119 2242 3178 420 124 1537 2278 408 +4,2 -31,5 -28,3 -2,7 Max. Frequency [MHz] Slices [#] LUTs [#] Power Dissip. [mW] 116 2462 2465 409 114 1666 1894 401 +1,7 -32,3 -12,2 -2 Synth. Quartus II SDMA Xilinx ISE III. Performance Assessment Model IDCT2D Use-Case: Required Hardware Components. TABLE II IDCT2D Use-Case: Synthesis on FPGAs. For ASIC synthesis we have adopted a 90nm CMOS technology. In Fig.3 the variation of the area of the CGRRA and the SDMA as the operating frequency is changed is shown. The longer critical paths, as in the FPGA exploration, led to a small frequency penalty (3% less in the CGRRA case). On the contrary, the benefit in terms of area is very promising (nearly the 40%). 500000,00 SDMA CGRRA 450000,00 400000,00 Area [µm2] the CGRR platform from high-level programs descriptions, as the proposed approach does. Kim et al. in [10] succeed in executing different applications on a fixed coarse-grained tile-based architecture composed of generic Processing Elements, rather the proposed framework allows the synthesis of a library of actor-specific optimized FUs as long as the definition of a customized application-specific dataflow-driven platform. Ogrenci et al. in [11] present a target-dependent approach, where no automatic derivation of the overall system is provided. Angermeier et al. in [12] propose a datafloworiented approach to reconfigurable systems design specifically addressing Partial Reconfiguration on FPGAs. This approach is target specific and FUs are not automatically derived. 350000,00 300000,00 250000,00 200000,00 655 665 675 685 695 705 715 725 Operating Frequency [MHz] Fig. 3. Implementation results on a 90nm technology. IV. Conclusion and perspectives In this paper we have proposed a novel design flow whose aim is two-fold. First, it contributes to close the gap between hardware specification and software development by automatically mapping complex software application on a given hardware platform. The proposed flow also IDCT1D model 1 IDCT1D model 2 IDCT1D CGRRA Fig. 2. The IDCT1D Use-Case. fulfils integration and specialization needs required by modern embedded systems design by leveraging on a reconfigurable hardware substrate. These goals are ensured by the combination of the OrccVHDL and the MDC tool. These tools are respectively able to provide efficient code generation using HLS from RVCCAL dataflow models to VHDL and to create a CGRR platform, extremely suitable for multi-purpose systems, relieving hardware developers from applications similarities analysis and composition. The proposed design paradigm is completely targetindependent. It allowed to achieve very promising results both on FPGAs and ASICs targets, where respectively the 30% and the 40% of area saving has been reached, at a negligible frequency penalty. More importantly exploiting the DPN MoC and combining the MDC tool with the Orcc-VHDL no users’ intervention are required in the design flow. Research directions in this field will regard mainly two other important hot-topics in embedded systems design: power management (exploiting the Sbox-based reconfiguration mechanism) and design space exploration (allowing an exchange of informations among Orcc-VHDL and the MDC). References [1] S. Carta, S. Pani, and L. Raffo, “Reconfigurable Coprocessor for Multimedia Application Domain,” Jorunal of VLSI Signal Processing Systems, vol. 44, no. 1-2, pp. 135–152, 2006. [2] V. Kumar and J. Lach, “Highly flexible multimode digital signal processing systems using adaptable components and controllers,” EURASIP J. Appl. Signal Process., vol. 2006, january. [3] M. Mattavelli, I. Amer, and M. Raulet, “The Reconfigurable Video Coding Standard [Standards in a Nutshell],” Signal Processing Magazine, IEEE, vol. 27, no. 3, pp. 159 –167, May 2010. [4] F. Palumbo, D. Pani, E. Manca, L. Raffo, M. Mattavelli, and G. Roquier, “RVC: A multi-decoder CAL Composer tool,” in IEEE international Conference on Design and Architectures for Signal and Image Processing (DASIP), 2010, pp. 144–151. [5] F. Palumbo, N. Carta, and L. Raffo, “The multi-dataflow composer tool: A runtime reconfigurable hdl platform composer,” in to be published in the IEEE international Conference on Design and Architectures for Signal and Image Processing (DASIP), 2011. [6] M. Wipliez, G. Roquier, and J.-F. Nezan, “Software Code Generation for the RVC-CAL Language,” Springer journal of Signal Processing Systems, 2009. [7] J. Gorin, M. Wipliez, J. Piat, F. Preteux, and M. Raulet, “An LLVM-based decoder for MPEG Reconfigurable Video Coding,” in IEEE international Workshop on Signal Processing Systems (SIPS), 2010. [8] P. Coussy and A. Morawiec, High-Level Synthesis: From Algorithm to Digital Circuit. Springer, 2008. [9] N. Siret, M. Wipliez, J. Nezan, and A. Rhatay, “Hardware code generation from dataflow programs,” in IEEE international Conference on Design and Architectures for Signal and Image Processing (DASIP), 2010, pp. 113 –120. [10] K. Yoonjin and R. Mahapatra, “Design Space Exploration for Efficient Resource Utilization in Coarse-Grained Reconfigurable Architecture,” IEEE Transactions on Very Large Scale Integration (VLSI) Systems, vol. 18, no. 10, pp. 1471–1482, oct. 2010. [11] S. Ogrenci Memik, E. Bozorgzadeh, R. Kastner, and M. Sarrafzadeh, “SPS: A strategically programmable system,” in Proceedings of the Int. Parallel and Distributed Processing Symposium (IPDPS), apr 2001. [12] J. Angermeier, S. Wildermann, E. Sibirko, and J. Teich, “Placing Streaming Applications with Similarities on Dynamically Partially Reconfigurable Architectures,” in Reconfigurable Computing and FPGAs (ReConFig), 2010 International Conference on, dec. 2010, pp. 91 –96.	28,603
https://openalex.org/W1999278752	OpenAlex	Open Science	CC-By	2,013	Molecular Dynamic Simulation to Explore the Molecular Basis of Btk-PH Domain Interaction with Ins(1,3,4,5)P4	Dan Lü	English	Spoken	7,419	14,787	Hindawi Publishing Corporation The Scientific World Journal Volume 2013, Article ID 580456, 10 pages http://dx.doi.org/10.1155/2013/580456 Hindawi Publishing Corporation The Scientific World Journal Volume 2013, Article ID 580456, 10 pages http://dx.doi.org/10.1155/2013/580456 Hindawi Publishing Corporation The Scientific World Journal Volume 2013, Article ID 580456, 10 pages http://dx.doi.org/10.1155/2013/580456 Academic Editors: P. Minkiewicz, Y. Muto, and W. A. Thompson Copyright © 2013 Dan Lu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Bruton’s tyrosine kinase contains a pleckstrin homology domain, and it specifically binds inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4), which is involved in the maturation of B cells. In this paper, we studied 12 systems including the wild type and 11 mutants, K12R, S14F, K19E, R28C/H, E41K, L11P, F25S, Y40N, and K12R-R28C/H, to investigate any change in the ligand binding site of each mutant. Molecular dynamics simulations combined with the method of molecular mechanics/Poisson-Boltzmann solvent- accessible surface area have been applied to the twelve systems, and reasonable mutant structures and their binding free energies have been obtained as criteria in the final classification. As a result, five structures, K12R, K19E, R28C/H, and E41K mutants, were classified as “functional mutations,” whereas L11P, S14F, F25S, and Y40N were grouped into “folding mutations.” This rigorous study of the binding affinity of each of the mutants and their classification provides some new insights into the biological function of the Btk-PH domain and related mutation-causing diseases. 1 Center for Systems Biology, Soochow University, Suzhou 215006, China 3 Department of Bioinformatics, Medical College, Soochow University, Suzhou 215123, China 3 Department of Bioinformatics, Medical College, Soochow University, Suzhou 215123, China Correspondence should be addressed to Guang Hu; huguang@suda.edu.cn Correspondence should be addressed to Guang Hu; huguang@suda.edu.cn Received 28 July 2013; Accepted 7 September 2013 Academic Editors: P. Minkiewicz, Y. Muto, and W. A. Thompson 2. Materials and Methods 2.1. Btk PH-Ins(1,3,4,5)P4 and Mutation Structures. The X- ray structure of the PH domain of Btk (PDB ID code: 2Z0P (A chain); resolution: 2.58 ˚A) [18], which is bound to Ins(1,3,4,5)P4, was used as the initial structure for all simu- lations. The PH domain has the common PH domain fold, which has a seven-stranded antiparallel 𝛽-sheet grouped into a 𝛽-sandwich and a 𝛼1 helix. For far distance away from the binding pocket, the Zn2+ ion in the structure was deleted for simplification and the forces between the Zn2+ ion and original four amino acids, His143 and Cys154/155/165, were manually added to reduce the flexibility of protein structure. Mutant structures (L11P, K12R, S14F, K19E, F25S, R28C/H, Y40N, E41K, and K12R-R28C/H) were obtained and visu- alized using the Sybyl software package (Tripos, St. Louis, MO). Ligand of Ins(1,3,4,5)P4 was extracted from the PH domain complex as an independent small molecule for the molecular dynamics (MD) and MM/PBSA simulation. All initial mutation structures were minimized by Sybyl using a distance-dependent dielectric function, with a nonbonded cut-off of 8 ˚A. Amber charges were assigned to the protein while Gasteiger-H¨uckel charges were given to Ins(1,3,4,5)P4. The whole system was minimized until no more atom colli- sions were found. According to previous studies, XLA-causing mutations can be mainly classified into two groups [24, 25]. The first group of mutations appears to prevent the formation of stable native-like structures, which are identified as “folding mutations.” In the second group named as “functional muta- tions,” the mutations do not affect the overall fold, though they may disrupt the ligand binding affinity leading to functional defects. Specially, K12 and R28 are significant in most functional mutations involved in the interaction with a ligand because a functionally based covariant pair is created, which contacts with a bound negatively charged ligand [26]. Generally, experimental studies have indicated the biological influence of amino acid mutation [24, 25, 27], but the proposed explanations do not exactly reveal much detailed molecular basis of the effects of amino acid mutations, such as a change in hydrogen bonds, binding sites, and binding free energy. In this paper, we have studied Btk-PH domain binding with the Ins(1,3,4,5)P4 structure in wild type and 11 well- known mutants [24, 25, 27], L11P, K12R, S14F, K19E, F25S, R28C/H, Y40N, E41K, and K12R-R28C/H. The method of molecular mechanics/Poisson-Boltzmann solvent-accessible surface area (MM/PBSA) was used to compare the binding affinity [28–32]. 2. Materials and Methods Three main aims were mainly raised and discussed in this study: (1) the classification of the eleven mutants, (2) the effects of amino acid mutations on the bio- logical function of the Btk-PH domain, and (3) the possible coordinate interactions of amino acid residues. Our simula- tions indicated that mutations K12R, K19E, R28C/H, E41K, and K12R-R28C/H were “functional mutations,” because neg- ative binding free energies were identified in the MM/PBSA calculation for these structures, whereas the others were considered to be “folding mutations” whose binding free energies are positive. The ligand clearly moves away from the binding pocket in R28C/H mutation structures. In K12R mutation structures, the ligand sites in the top of the binding Pocket, which results in the evident decrease of binding free energy. Thus, the biological function of Btk is weakened in all three mutant structures. The K19E mutation leads to a major change in the 𝛽1-𝛽2 loop, leading to weak membrane targeting and the suppression of Btk activation. The E41K mutation increases the positive charge of the 𝛽3-strand, which increases the positively charged surface. This may provide a cavity for binding with another ligand, which could enhance the membrane targeting and activation of Btk. Remarkably, the two double amino acids mutant structures K12R-R28C/H retain a similar ligand binding affinity to the wild type structure, because new coordinate interactions by the amino acid pairs R12 and K53 are found to play important roles in the binding with Ins(1,3,4,5)P4. We hope that our 2.2. MD Simulation. Before the MD simulation, the charge information of Ins(1,3,4,5)P4 was calculated using the RESP method [33] encoded in the AMBER suite program (version 9) [34], followed by Gaussian 03 for calculations at the Hartree-Fock (HF)/6-31G∗level. After the application of the AMBER03 force field to proteins [35] and the general AMBER force field (gaff) to ligands [36], each complex was loaded into the AMBER9 program, adding Na+ ions with a 1 ˚A grid to neutralize the system. Finally, an 8 ˚A water TIP3PBOX was loaded into the system to form a complex environment. Subsequently, energy minimization was performed to remove any inappropriate contacts. 1. Introduction critical for signal transmission. Several missense mutations in the PH domain have been widely studied and so far con- sidered to be the only known cause of the disease [12]. The PH domain is responsible for binding with phosphatidylinositols, showing the importance for the regulation of membrane targeting. Thus, a mutation in the PH domain can influence the binding affinity with a ligand, membrane targeting, and the activation of Btk [13, 14]. Of the various phos- phatidylinositols, the Btk-PH domain has higher specificity and binding affinity with inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4) [15–17]. The crystal structure of a complex of the Btk-PH domain with Ins(1,3,4,5)P4 (PDB ID code: 2Z0P [18]) shows that the Btk-PH domain recognizes Ins(1,3,4,5)P4 in a canonical manner [19, 20].h Bruton’s tyrosine kinase (Btk) is a member of the Tec family of kinases and the only known one associated with human disease [1, 2]. Previous studies have indicated the significance of Btk in B-cell development, differentiation, and signaling [3, 4]. Once the Btk-dependent signal transduction pathway is inactivated, B cells remain at the pre-B-cells stage leading to X-linked agammaglobulinemia (XLA) in humans, which is one of the most frequently inherited immunodeficient dis- orders in human, and X-linked immunodeficiency (Xid) in mice [5–8]. The Btk protein contains Src-homology 2 and 3 domains (SH2 and SH3), a catalytic SH1 domain, a Tec-homology (TH) domain, and an N-terminal pleckstrin homology (PH) domain [9–11]. Studies have shown that XLA mutations in Btk can be mapped to all five domains of the kinase, which are The PH domain is a structural protein domain contain- ing approximately 120 amino acid residues that retains a highly conserved three-dimensional organization of different The Scientific World Journal 2 findings help to explain the relationship between residue mutations and biological function, as well as the molecular basis of related diseases. proteins, despite their poorly conserved primary sequences [21–23]. The core structure is a 𝛽-sandwich of two almost orthogonal 𝛽-sheets consisting of three and four strands, respectively. The opposite edge of the structure is capped by an amphipathic C-terminal 𝛼-helix. Six loops connect the 𝛽-strands, while the amino acids in the 𝛽1-𝛽2 and 𝛽3-𝛽4 loops form a hydrogen bond network that interacts with Ins(1,3,4,5)P4 [19]. 2. Materials and Methods Binding free energy (Δ𝐺bind) of ligand on Btk-PH domain was calcu- lated by single trajectory method of MM/PBSA, according to the following equation: der Waals energy (𝐸vdw), which were calculated using the same force field as that of MD simulations with no cut- off. Notably, the 𝐸int was assigned to zero in (1), since the torsion angle energies of the complex and the separated part were calculated from the same trajectory. In the MM/PBSA calculation, 2000 snapshots of each model were extracted from the last 10 ns of the trajectories at time intervals of 5 ps. Δ𝐺bind = 𝐺complex −𝐺PH −𝐺li, (1) (1) where the free energies of complex, PH domain, and ligand are denoted as 𝐺complex, 𝐺PH, and 𝐺li, respectively. Free energy (𝐺) was calculated according to following equations: 2. Materials and Methods MD simulations were conducted with a nonbonded cut- off of 8 ˚A, the dielectric constant of 1.0, integration step of a 2 fs time interval, and Particle Mesh Ewald (PEM) for long-range electrostatic interaction calculation [37], using the following protocol: (1) 500 ps of heating equilibration with weak restraints on each complex in Cartesian space using a harmonic potential, where the temperature was gradually increased to 300 K, with constant volume, and Langevin dynamics for temperature control; (2) 500 ps of density equi- libration, 300 K thermal bath at 1.0 atm of pressure (atm = 101.3 kPa) periodic boundary conditions with isotropic posi- tion scaling, and Langevin dynamics for temperature control; (3) 1 ns of constant pressure equilibration with weak restraint on the ligand and 20 ns of constant pressure equilibration at 300 K. 3 3 The Scientific World Journal 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 4 2 1 0 3 2 1 0 K12RR28H K12RR28C E41K K19E K12R WT Y40N F25S S14F L11P R28H R28C RMSD (angstrom) RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) Figure 1: Time dependence of the root-mean-square deviations (RMSDs) for the C𝛼atoms from their initial structure of 20 ns MD simulations of all complex structures. 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 K12RR28H K12RR28C E41K K19E K12R WT RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 3 2 1 0 3 2 1 0 3 2 1 0 3 2 1 0 3 4 2 1 0 3 2 1 0 Y40N F25S S14F L11P R28H R28C RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) K12RR28C RMSD (angstrom) S14F L11P RMSD (angstrom) RMSD (angstrom) Figure 1: Time dependence of the root-mean-square deviations (RMSDs) for the C𝛼atoms from their initial structure of 20 ns MD simulations of all complex structures. he root-mean-square deviations (RMSDs) for the C𝛼atoms from their initial structure of 20 ns MD simulations 2.3. MM-PBSA Calculations for Binding Free Energy. 3. Results and Discussions 4 K12RR28H K12RR28C E41K K19E K12R WT RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 The Scientific World Journal 4 The Scientific World Journal Y40N F25S S14F L11P R28H R28C RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 6 4 2 0 6 4 2 0 6 8 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 Y40N F25S S14F L11P RMSD (angstrom) RMSD (angstrom) Figure 2: Time dependence of the root-mean-square deviations (RMSDs) for the ligand from its initial structure of 20 ns MD simulations of all complex structures. the protein and the ligand. Meanwhile, the RMSD value of the ligand Ins(1,3,4,5)P4 was also calculated to further indicate the stability of the structures. In Figure 2, it was obvious that the ligand of WT, K12R, K19E, E41K, K12R-R28C, and K12R-R28H was more stable than that of R28C, R28H, L11P, S14F, F25S, and Y40N. Therefore, it was believed that R28C, R28H, L11P, S14F, F25S, and Y40N mutants might share some similar instabilities, which was in good accordance with the predefined classifications based on experimental results [27].fi will result in the loss of function of the PH domain com- pletely, although S14F was considered to be a “functional mutation” in an earlier study [24, 25]. On the contrary, the other 7 mutations have negative binding free energies. With the stable RMSD values, it is proposed that the mutations of K12R, K19E, R28C/H, E41K, and K12R-R28C/H may be considered as “functional mutations,” which do not affect the overall fold but can be expected to disrupt the binding affinities between the protein and the ligand [24, 27]. In this paper, the binding free energies of seven mutation structures were carefully compared to the wild type, using Δ𝐺bind as the criterion. Different mutational simulations provided impor- tant information about the affinity drift of the ligand-binding protein complex, which were further discussed below. ii In order to offer a better criterion for classification of the mutants, the MM/PBSA method was used to estimate the binding affinities of the PH domain and the ligand in the 12 structures. Energy results for all the MM/PBSA calculations shown in Table 1 indicated that these mutations could be clearly divided into two groups. 3. Results and Discussions MD simulations were used to investigate the complexes of 11 mutants and wild type PH domain and the interaction behav- ior between the PH domain and the ligand Ins(1,3,4,5)P4. Details of the simulation setup are provided in Materials and Methods. The root-mean-square deviation (RMSD) values of the C𝛼atoms relative to the initial coordinates were calcu- lated using the 20 ns trajectory data. As shown in Figure 1, the RMSD tended to be flat after 5 ns’ simulation in WT, K12R, K19E, E41K, K12R-R28C, K12R-R28H, R28C, and R28H at about 2 ˚A, indicating stable conformations. As for the RMSD values for other four mutants, relative large fluctuations were observed during the 20 ns’ simulation, suggesting unsta- ble structures and maybe poor binding affinities between 𝐺= 𝐸gas + 𝐺sol −𝑇𝑆config, 𝐸gas = 𝐸vdw + 𝐸ele + 𝐸int, 𝐺sol = 𝐺elec + 𝐺SA, (2) where 𝐸gas defines the molecular mechanical energy, 𝐺sol defines a solvation free energy, which is further decomposed to polar (𝐺elec) and nonpolar (𝐺SA) terms, and 𝑆config defined as configurational entropy, which is normally derived from normal model analysis. 𝐸gas was the sum of contributions from internal energies including bond, angle, and torsion angle energies (𝐸int), electrostatic energy (𝐸ele), and van 4 The Scientific World Journal K12RR28H K12RR28C E41K K19E K12R WT Y40N F25S S14F L11P R28H R28C RMSD (angstrom) RMSD (angstrom) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 0.0 4.0 8.0 12.0 16.0 20.0 Time (ns) 6 4 2 0 6 4 2 0 6 8 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 6 4 2 0 Figure 2: Time dependence of the root-mean-square deviations (RMSDs) for the ligand from its initial structure of 20 ns MD simulations of all complex structures. 3. Results and Discussions It was convinced that the “folding mutations” could not fold into a stable native-like structure to perform the function of the PH domain [27]. In our results, it is obvious that the four mutations, L11P, S14F, F25S, and Y40N, own a positive binding free energy, though the enthalpy change is favorable, implying the formation of complex is not allowed by thermodynamics. As aforemen- tioned, these four mutations also had unstable RMSD value. Therefore, it is most likely that the four “folding mutations” 3.1. Structural and Binding Affinity Analysis of “Functional Mutations”. According to the MM/PBSA simulations, the binding affinity of the 7 “functional mutations” was ranked to offer a better understanding of risks in disease, which was displayed in Table 1. 3.1. Structural and Binding Affinity Analysis of “Functional Mutations”. According to the MM/PBSA simulations, the binding affinity of the 7 “functional mutations” was ranked to offer a better understanding of risks in disease, which was displayed in Table 1. As aforementioned, the RMSD values of the ligand in R28C and R28H mutants were greater than 4, which was two times larger than that of wild type. Therefore, it is most The Scientific World Journal 5 Table 1: Ranked calculated binding free energies (kcal/mol) using MM/PBSA for the twelve protein-ligand systems. 3. Results and Discussions The mutations removed the hydrogen bond interaction between R28 and the ligand while forming new hydrogen bond interactions with residues on the 𝛽4 strand (K53) and 𝛽1-𝛽2 loop (K17), respectively, which would not provide suitable side chains to bind with the ligand. Accordingly, it was considered that the R28C/H mutations had greatly reduced the binding affinity with the ligand. And the biological function of the two mutants was severely weakened, which may be considered as a primary factor that leads to associated diseases such as XLA and Xid [5]. likely that the ligand completely moves away from the original binding pocket of the PH domain and this was consistent with the result of the binding free energy calculation for R28C/H mutants (Table 1). It is clear that the neutral charged cysteine has a sulfur atom in its side chain, which imposes a larger steric restriction compared with arginine. Although it has the same positive charge as arginine, histidine has a five-member ring that produces more steric hindrance than the side chain of arginine. Structural analysis in Figure 3 verified that the ligand was already out of the binding pocket. The mutations removed the hydrogen bond interaction between R28 and the ligand while forming new hydrogen bond interactions with residues on the 𝛽4 strand (K53) and 𝛽1-𝛽2 loop (K17), respectively, which would not provide suitable side chains to bind with the ligand. Accordingly, it was considered that the R28C/H mutations had greatly reduced the binding affinity with the ligand. And the biological function of the two mutants was severely weakened, which may be considered as a primary factor that leads to associated diseases such as XLA and Xid [5]. g g Structural analysis of the wild type showed that K12, which was located at the bottom of the binding pocket, formed hydrogen bonds with the 3- and 4-phosphates of the ligand, which is critical for the twist of 𝛽1-𝛽2 loops [24]. In the K12R mutant, there was obvious decrease in binding free energy in the MM/PBSA simulation (Table 1), which could be explained by the difference in the conformation and the change in the hydrogen bond interactions of the ligand. Sharing the same positive change as lysine, the three atoms in arginine tail formed a conjugated system, which decreased the flexibility and increased the steric hindrance of this residue, leading to the conformational change required for ligand binding. 3. Results and Discussions ELE VDW PB SA PBTOT 𝑇Δ𝑆 Δ𝐺bind WT −3031.77 (9.01a) −0.16 (0.07) 2961.60 (7.63) −4.28 (0.47) −74.61 (5.10) −45.29 (1.43) −29.32 (1.49) K12R-R28C −3119.66 (10.33) −0.12 (0.05) 3044.04 (8.61) −4.25 (0.18) −79.99 (3.37) −52.50 (1.45) −27.49 (1.06) K19E −3143.08 (8.92) −0.17 (0.03) 3069.08 (7.38) −4.21 (0.16) −78.38 (5.33) −51.09 (1.60) −27.29 (0.73) K12R-R28H −3029.46 (8.74) −0.05 (0.02) 2962.73 (6.43) −4.12 (0.21) −73.30 (2.02) −51.66 (1.37) −21.64 (0.66) E41K −2855.47 (7.73) −0.28 (0.16) 2789.39 (5.95) −4.16 (0.23) −70.52 (3.12) −50.08 (1.52) −20.44 (0.38) K12R −2922.34 (9.17) −0.08 (0.03) 2859.80 (8.38) −4.30 (0.21) −66.92 (3.62) −52.84 (1.57) −14.08 (0.94) R28C −2612.17 (5.95) −1.90 (0.23) 2552.82 (5.03) −3.69 (0.16) −64.94 (1.87) −57.28 (2.47) −7.66 (0.03) R28H −2591.08 (6.16) −1.61 (0.17) 2533.38 (4.65) −3.70 (0.17) −63.01 (1.16) −56.80 (2.54) −6.21 (0.02) S14F −2573.79 (7.06) −3.37 (0.74) 2545.38 (6.99) −3.42 (0.25) −35.20 (2.47) −57.49 (3.02) 22.29 (0.98) L11P −2599.24 (7.61) −1.75 (0.41) 2578.10 (6.70) −3.94 (0.33) −26.83 (1.42) −58.75 (4.47) 31.92 (2.02) Y40N −2550.35 (7.0) −2.57 (0.59) 2531.48 (6.23) −3.92 (0.27) −25.36 (1.84) −58.34 (3.89) 32.98 (1.68) F25S −2501.74 (7.17) −2.59 (0.69) 2487.39 (6.64) −3.77 (0.19) −20.71 (1.54) −58.32 (4.15) 37.61 (1.82) aThe statistical error was estimated on the basis of the deviation between block averages. ELE: electrostatic energy; VDW: van der Waals energy; PB: the PB solvation energy; SA: the surface area energy (nonpolar solvation energy); PBTOT: sum of ELE, VDW, PB, and SA. The absolute temperature (𝑇) was set to 300 K in the MM-PBSA calculations. Δ𝐺bind: subtracting 𝑇Δ𝑆from PBTOT. d calculated binding free energies (kcal/mol) using MM/PBSA for the twelve protein-ligand systems. ligand-conformational changes due to hydrogen bond rup- ture. The time evolution of the root-mean-square fluctuation (RMSF) from the initial structure was also analyzed to further evaluate the ligand binding behavior of these mutations. likely that the ligand completely moves away from the original binding pocket of the PH domain and this was consistent with the result of the binding free energy calculation for R28C/H mutants (Table 1). It is clear that the neutral charged cysteine has a sulfur atom in its side chain, which imposes a larger steric restriction compared with arginine. Although it has the same positive charge as arginine, histidine has a five-member ring that produces more steric hindrance than the side chain of arginine. Structural analysis in Figure 3 verified that the ligand was already out of the binding pocket. 3. Results and Discussions The hydrogen bond with the 3-phosphates of the ligand was also fully ruptured in K12R mutant, resulting in a loss of specificity in the Btk-PH domain for the Ins(1,3,4,5)P4 ligand. As shown in Table 2, the K12R mutant abolished the hydrogen bond network formed by K12, R28, and the ligand; thereby the contribution of the conserved covariation pair on the biological function of the Btk-PH domain was removed. As shown in Table 1, according to the MM/PBSA simu- lations, the free binding free energies of the mutants K19E, E41K, and K12R-R28C/H were similar to the wild type, whereas the K12R mutant was relatively smaller (−14.08 kcal/ mol). Furthermore, compared with the wild type, the ligand was still located in the binding pocket of these mutants, as shown in Figures 3(d)–3(h), although there were different In the wild type, as shown in Figure 3(a), K19 contacts D148 via a hydrogen bond, which moves the 𝛽1-𝛽2 loop closer to the tail of the structure and moves it away from the 6 The Scientific World Journal 6 (a) (b) (c) (d) (e) (f) (g) (h) The snapshot structures taken at time 20 ns of wild type and all “functional mutations” complexes from MD simula nding important residues are labeled and shown in stick, while hydrogen bonds are displayed in yellow dashed li ex; (b) R28C mutation complex; (c) R28H mutation complex; (d) K12R mutation complex; (e) K19E mutation comp mplex; (g) K12R-R28C mutation complex; (h) K12R-R28H mutation complex. (a) (b) (c) (d) (a) (b) (a) (c) (d) (c) (d) (f) (f) (h) (e) (f) (h) (e) (g) ( ) (g) (h) (g) Figure 3: The snapshot structures taken at time 20 ns of wild type and all “functional mutations” complexes from MD simulation. Ligands and surrounding important residues are labeled and shown in stick, while hydrogen bonds are displayed in yellow dashed lines. (a) Wild type complex; (b) R28C mutation complex; (c) R28H mutation complex; (d) K12R mutation complex; (e) K19E mutation complex; (f) E41K mutation complex; (g) K12R-R28C mutation complex; (h) K12R-R28H mutation complex. Figure 3: The snapshot structures taken at time 20 ns of wild type and all “functional mutations” complexes from MD simulation. Ligands and surrounding important residues are labeled and shown in stick, while hydrogen bonds are displayed in yellow dashed lines. 3. Results and Discussions 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) WT E41K K19E 𝛽1-𝛽2 loop 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) WT E41K K19E 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) K12R K12RR28H K12RR28C 𝛽1-𝛽2 loop 𝛽1-𝛽2 loop Figure 4: Residue fluctuations obtained by average residual fluctuations between residues 1 and 70 for six simulations, wild type (WT), K19E, d h l ll d l d l Th d f 𝛽𝛽l h hl h d 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) K12R K12RR28H K12RR28C 𝛽1-𝛽2 loop RMSF (angstrom) RMSF (angstrom) K12RR28C Figure 4: Residue fluctuations obtained by average residual fluctuations between residues 1 and 70 for six simulations, wild type (WT), K19E, E41K, K12R, and K12R-R28C/H, over the 20 ns simulations are illustrated in solid lines. The residues of 𝛽1-𝛽2 loop are highlighted. ligand. In the mutant K19E, a stable hydrogen bond network is formed among S14, T20, and the ligand after the rupture of the hydrogen bond between E19 and D148 (Table 2), which leads to a higher binding free energy with the ligand than the wild type while leading to an almost 90∘change in the direction of the 𝛽1-𝛽2 loop, as shown in Figure 3(e). The RMSF analysis in Figure 4 also confirmed that the 𝛽1-𝛽2 loop had much ligand. In the mutant K19E, a stable hydrogen bond network is formed among S14, T20, and the ligand after the rupture of the hydrogen bond between E19 and D148 (Table 2), which leads to a higher binding free energy with the ligand than the wild type while leading to an almost 90∘change in the direction of the 𝛽1-𝛽2 loop, as shown in Figure 3(e). The RMSF analysis in Figure 4 also confirmed that the 𝛽1-𝛽2 loop had much greater fluctuations than the wild type. 3. Results and Discussions (a) Wild type complex; (b) R28C mutation complex; (c) R28H mutation complex; (d) K12R mutation complex; (e) K19E mutation complex; (f) E41K mutation complex; (g) K12R-R28C mutation complex; (h) K12R-R28H mutation complex. 7 The Scientific World Journal Table 2: Comparison of hydrogen bond interactions of five “functional mutations” and wild-type (WT) structures. Only hydrogen bonds with occupation ratio of the trajectory greater than 30%, atomic distance of two heavy atoms smaller than 3 ˚A, and atomic angle smaller than 60∘are presented. The rank of amino acid according to its occupation ratio of the trajectory is from large to small. ph g p j y g WT E41k K19E K12R K12R-R28H K12R-R28C Arg28 3HB with 3-PGa Lys12 3HB with 3-PG Ser14 1HB with 4-PG Lys26 4HB with 1-PG Ser 21 2HB with 5-PG 1HB with 6-OH𝛽 Gln15 1HB with 4-PG Lys17 2HB with 4-PG Lys53 2HB with 3-GP Arg28 3HB with 3-PG Lys12 3HB with 3-PG Ser 14 2HB with 4-PG Ser 21 2HB with 5-PG Gln15 1HB with 4-PG Lys26 2HB with 1-PG Lys53 2HB with 3-GP Arg28 3HB with 3-PG Lys12 3HB with 3-PG Ser 14 1HB with 4-PG Thr 20 1HB with 5-PG Lys17 2HB with 4-PG Gln15 1HB with 4-PG Ser 21 2HB with 5-PG 1HB with 6-OH Lys26 2HB with 1-PG Lys53 2HB with 3-PG Arg12 2HB with 4-PG Ser 21 1HB with 5-PG Lys17 2HB with 5-PG Ser 14 1HB with 4-PG Gln15 1HB with 4-PG Lys26 1HB with 1-PG Arg12 3HB with 3-PG Ser 14 1HB with 4-PG Lys53 2HB with 3-PG Gln15 3HB with 4-PG Lys17 2HB with 4-PG Ser 21 1HB with 5-PG 1HB with 6-OH Lys26 1HB with 1-PG Arg12 3HB with 3-PG Lys53 2HB with 3-GP Ser 14 1HB with 4-PG Gln15 3HB with 4-GP Lys17 2HB with 4-PG Ser 21 2HB with 5-PG 1HB with 6-OH Lys26 1HB with 1-PG aHB and PG represent hydrogen bond and phosphate group, respectively. 𝛽OH represents OH group in the inositol ring. 3. Results and Discussions 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) WT E41K K19E 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 Residues RMSF (angstrom) K12R K12RR28H K12RR28C 𝛽1-𝛽2 loop 𝛽1-𝛽2 loop Figure 4: Residue fluctuations obtained by average residual fluctuations between residues 1 and 70 for six simulations, wild type (WT), K19E, E41K, K12R, and K12R-R28C/H, over the 20 ns simulations are illustrated in solid lines. The residues of 𝛽1-𝛽2 loop are highlighted. ligand. In the mutant K19E, a stable hydrogen bond network is greater fluctuations than the wild type. Therefore, it is possible WT E41k K19E K12R K12R-R28H K12R-R28C Arg28 3HB with 3-PGa Lys12 3HB with 3-PG Ser14 1HB with 4-PG Lys26 4HB with 1-PG Ser 21 2HB with 5-PG 1HB with 6-OH𝛽 Gln15 1HB with 4-PG Lys17 2HB with 4-PG Lys53 2HB with 3-GP Arg28 3HB with 3-PG Lys12 3HB with 3-PG Ser 14 2HB with 4-PG Ser 21 2HB with 5-PG Gln15 1HB with 4-PG Lys26 2HB with 1-PG Lys53 2HB with 3-GP Arg28 3HB with 3-PG Lys12 3HB with 3-PG Ser 14 1HB with 4-PG Thr 20 1HB with 5-PG Lys17 2HB with 4-PG Gln15 1HB with 4-PG Ser 21 2HB with 5-PG 1HB with 6-OH Lys26 2HB with 1-PG Lys53 2HB with 3-PG Arg12 2HB with 4-PG Ser 21 1HB with 5-PG Lys17 2HB with 5-PG Ser 14 1HB with 4-PG Gln15 1HB with 4-PG Lys26 1HB with 1-PG Arg12 3HB with 3-PG Ser 14 1HB with 4-PG Lys53 2HB with 3-PG Gln15 3HB with 4-PG Lys17 2HB with 4-PG Ser 21 1HB with 5-PG 1HB with 6-OH Lys26 1HB with 1-PG Arg12 3HB with 3-PG Lys53 2HB with 3-GP Ser 14 1HB with 4-PG Gln15 3HB with 4-GP Lys17 2HB with 4-PG Ser 21 2HB with 5-PG 1HB with 6-OH Lys26 1HB with 1-PG aHB and PG represent hydrogen bond and phosphate group, respectively. 𝛽OH represents OH group in the inositol ring. aHB and PG represent hydrogen bond and phosphate group, respectively. 𝛽OH represents OH group in the inositol ring. 3. Results and Discussions Despite the higher steric hindrance after the substitution of glutamate with lysine, the ligand slightly shifts to the 𝛽1-𝛽2 loop in Figure 3(f), which results in a reduced flexibility of the 𝛽1-𝛽2 loop (RMSF in Figure 4). The binding free energy of the E41K mutant was also smaller than the wild type, as shown in Table 1. Thus, E41K may retain most of the biological functionality of the wild type. However, the mutation in E41K increased the positively charged complex surface of the region located above the 𝛽3 and 𝛽4 strand, which was favorable for interaction with the negatively charged membrane surface, while it also offered a second ligand binding site. Therefore, it is likely that the E41K mutant may increase the activation of Btk, which is known as a gain of function [24, 27]. 3.2. Covariant Pair Identification in K12R-R28C/H Mutations. Of the disease-causing mutations, the mutations in K12 and R28 always coexist with other mutations, although with lower frequencies, which may be due to the diversity of the PH domain. Single mutations of K12 and R28 have been known to weaken the biological function of the Btk-PH domain to different degrees, while it is also reported that K12 and R28 can form a conserved covariant pair that maintains the bind- ing affinity of Ins(1,3,4,5)P4, which plays an important role in the biological function of the Btk-PH domain [26]. Therefore, the double residue mutations of K12R-R28C/H were closely analyzed to underly the molecular basis. Compared with the wild type structure, the ligand was still located in the bottom of the binding pocket of the two mutations (Figures 3(g) and 3(h)). The MM/PBSA simulations indicated a similar free binding energy for K12R-R28H and K12R-R28C (Table 1). This could be explained by the formation of a new covariant pair between R12 and K53 as K12-R28. Unlike the K12R mutation, the hydrogen bond network of 3-phosphates was kept through R12 and K53 in the mutation of K12R-R28C/H, which ensured the preservation of ligand specificity, whereas all hydrogen bond interactions between H28/C28 and lig- and were ruptured (Table 2). Interestingly, a new conserved hydrogen bond was discovered between K53 and the ligand during the MD simulations. 3. Results and Discussions Therefore, it is possible the significant change in the 𝛽1-𝛽2 loop of this mutation could eliminate the positively charged surface that interacts with the negatively charged membrane surface, resulting in poor membrane targeting and incomplete activation of Btk. This may mean that the biological function of Btk is not fully available. Therefore, K19E was also a loss-of-function The Scientific World Journal 8 mutation, although it had a similar binding affinity with the ligand than the wild type, which was in good agreement with the experimental results [24, 27]. Y40N were considered to be “folding mutations,” whereas mutants K12R, K19E, R28C/H, E41K, and K12R-R28C/H were classified into “functional mutations.” The effects of “functional mutations” on the biological function of the Btk-PH domain have also been predicted. In the R28C/H mutation, the ligands completely left the binding pocket and formed new interactions with the 𝛽1-𝛽2 loop and the 𝛽4 strand, respectively. For the K12R mutation, the biological function of the Btk-PH domain may be reduced by the lack of hydrogen bond interaction with 3-phosphates when the ligand is formed, which determines the specificity of the Btk- PH domain. As for the K19E mutation, the change in the residue charge and the trend of the 𝛽1-𝛽2 loop eliminated the positively charged surface that interacted with the negatively charged membrane surface, resulting in poor membrane targeting and incomplete activation of Btk. In contrast, E41K increased the positive surface of the Btk-PH domain, thereby increasing the possibility of the binding with a second ligand, which may result in a gain of function. Of note, the K12R- R28C/H mutant retained a high binding affinity without the displacement of the ligand from the binding pocket. A new odd positively charged pair was formed between R12 and K53 in the K12R-R28C/H mutation, which was similar to the K12-R28 in the wild type. This formation may interact with a bound negatively charged inositol phosphate, which may preserve the biological function of the Btk-PH domain and this should be further investigated experimentally. Compared with K19E, the mutant of E41K has the opposite effect on the biological function of Btk. As shown in Figure 3(a), E41 was located in the 𝛽3 strand with some distance from the ligand, but it is known to have great importance in the contact with the membrane. 3. 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https://openalex.org/W3041898132	OpenAlex	Open Science	CC-By	2,020	CCR2 of Tumor Microenvironmental Cells Is a Relevant Modulator of Glioma Biology	Matthäus Felsenstein	English	Spoken	13,926	27,004	Received: 27 June 2020; Accepted: 10 July 2020; Published: 13 July 2020 Received: 27 June 2020; Accepted: 10 July 2020; Published: 13 July 2020 Abstract: Glioblastoma multiforme (GBM) shows a high inﬂux of tumor-associated macrophages (TAMs). The CCR2/CCL2 pathway is considered a relevant signal for the recruitment of TAMs and has been suggested as a therapeutic target in malignant gliomas. We found that TAMs of human GBM specimens and of a syngeneic glioma model express CCR2 to varying extents. Using a Ccr2-deﬁcient strain for glioma inoculation revealed a 30% reduction of TAMs intratumorally. This diminished immune cell inﬁltration occurred with augmented tumor volumes likely based on increased cell proliferation. Remaining TAMs in Ccr2-/- mice showed comparable surface marker expression patterns in comparison to wildtype mice, but expression levels of inﬂammatory transcription factors (Stat3, Irf7, Cox2) and cytokines (Ifnβ, Il1β, Il12α) were considerably aﬀected. Furthermore, we demonstrated an impact on blood vessel integrity, while vascularization of tumors appeared similar between mouse strains. The higher stability and attenuated leakiness of the tumor vasculature imply improved sustenance of glioma tissue in Ccr2-/- mice. Additionally, despite TAMs residing in the perivascular niche in Ccr2-/- mice, their pro-angiogenic activity was reduced by the downregulation of Vegf. In conclusion, lacking CCR2 solely on tumor microenvironmental cells leads to enhanced tumor progression, whereby high numbers of TAMs inﬁltrate gliomas independently of the CCR2/CCL2 signal. Keywords: tumor-associated macrophages (TAMs), blood vessel integrity; tumor angiogenesis; CCR2/CCL2 signaling; GBM CCR2 of Tumor Microenvironmental Cells Is a Relevant Modulator of Glioma Biology Matthäus Felsenstein 1,†, Anne Blank 1, Alexander D. Bungert 1, Annett Mueller 1, Adnan Ghori 1, Irina Kremenetskaia 1, Olga Rung 1, Thomas Broggini 1,†, Kati Turkowski 1, Lea Scherschinski 1, Jonas Raggatz 1, Peter Vajkoczy 1,2,,‡ and Susan Brandenburg 1,‡ 1 Department of Experimental Neurosurgery Charité, Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, Germany; matthaeus.felsenstein@charite.de (M.F.); anne.blank@charite.de (A.B.); alexander.bungert@charite.de (A.D.B.); am2374@medschl.cam.ac.uk (A.M.); adnan.ghori@charite.de (A.G.); irina.kremenetskaia@charite.de (I.K.); olga.rung@web.de (O.R.); thomas.broggini@gmail.com (T.B.); kati.turkowski@mpi-bn.mpg.de (K.T.); lea.scherschinski@charite.de (L.S.); jonas.raggatz@outlook.com (J.R.); susan.brandenburg@charite.de (S.B.) g 2 Department of Neurosurgery Charité, Universitätsmedizin Berlin, 10117 Berlin, Germany Correspondence: peter.vajkoczy@charite.de; Tel.: +49-30-450-560-002 p p j y † Present address: Department of Surgery, Charité—Universitätsmedizin Berlin, 10117 Berlin, Germany (M.F.); Department of Physics University of California at San Diego La Jolla CA 92093 USA (T B ) † Present address: Department of Surgery, Charité—Universitätsmedizin Berlin, 10117 Berlin, Germany (M Department of Physics, University of California at San Diego, La Jolla, CA 92093, USA (T.B.). ‡ Peter Vajkoczy and Susan Brandenburg contribute equally to this work. ‡ Peter Vajkoczy and Susan Brandenburg contribute equally to this work. Received: 27 June 2020; Accepted: 10 July 2020; Published: 13 July 2020 Received: 27 June 2020; Accepted: 10 July 2020; Published: 13 July 2020 www.mdpi.com/journal/cancers cancers cancers cancers Article Keywords: tumor-associated macrophages (TAMs), blood vessel integrity; tumor angiogenesis; CCR2/CCL2 signaling; GBM cancers cancers 1. Introduction Glioblastoma multiforme (GBM) is a poorly diﬀerentiated human brain tumor entity (WHO IV◦) and displays markedly aggressive and invasive features. GBM tumors account for 60–70% of all malignant gliomas and have high recurrence rates with a dismal prognosis [1,2]. Large numbers of non-dysplastic cells belong to the glioma mass [3], including tumor-associated macrophages (TAMs) which make up 5–30% of all cells within the tumor compartment [4,5]. Cancers 2020, 12, 1882; doi:10.3390/cancers12071882 www.mdpi.com/journal/cancers 2 of 19 Cancers 2020, 12, 1882 TAMs inﬁltrate due to chemotactic signals [6,7]. For example, CCL2 was shown to be released from glioma cells [5,8]. Its chemokine (C–C motif) receptor 2 (CCR2) is mainly located on the surface of TAMs [9,10]. The impact of this interaction has been demonstrated in several brain pathologies such as in atherosclerosis, autoimmune encephalitis, or even post-stroke inﬂammatory condition [11–14]. Furthermore, overexpression of CCL2 in glioblastoma rodent models showed that CCL2 increased the amount of inﬁltrating TAMs [15,16]. An anti-CCL2-antibody increased the survival time and reduced the number of TAMs [17]. Targeting the CCR2/CCL2 pathway showed eﬀectiveness in other tumor models as it suppresses tumor progression [18–20]. Hence, the CCR2/CCL2 axis has been identiﬁed as an interesting target for tumor therapy, and collected data indicate that CCR2/CCL2 signaling plays a relevant role in the tumor-myeloid cell-interaction. Yet, the precise function of CCR2-signaling for glioma progression as well as angiogenesis has not been elucidated. Previously, the function of CCR2/CCL2 signal for glioma biology was investigated by focusing on its ligand CCL2. Both, Ccl2-/- mice and anti-CCL2 antibodies were investigated [17,21,22]. In our study, we used Ccr2-/- mice [23] in combination with the GL261 immunocompetent mouse glioma model to evaluate the importance of the CCR2 expression on the tumor microenvironment, especially on TAM recruitment and their phenotype as well as glioma vascularization. We demonstrate an impaired inﬂux of myeloid cells in Ccr2-deﬁcient animals, whereby remaining TAMs displayed a varied expression of pro-inﬂammatory molecules. Ultimately, we observed enhanced tumor growth and an increase in tumor blood vessel integrity. Using the Ccr2-/- knockout model, severe glioma microenvironmental alterations were uncovered, which do not appear beneﬁcial. 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissue It has been demonstrated that CCR2/CCL2 signaling plays a pivotal role in chemo-attraction during neuro-inﬂammatory processes [15,24]. Analyzing the expression level of CCR2 within GBM patient samples using the TCGA database, we found that 51.5% of all patient specimens show regulated CCR2 expression (19.3% up-regulation and 32.2% down-regulation; Figure 1a). Additionally, we investigated surgically resected human brain tissues at our institution and observed CCR2 positive staining both in epilepsy (EP) (3/3) as well as GBM tissue samples (6/6) revealing variable expression levels in glioma specimens (Figure 1b). Moreover, we detected CCR2 expression in freshly isolated myeloid cells (CD11b+) derived from corresponding human brain specimens. CD11b+ cells of GBM samples predominantly displayed down-regulation with respect to myeloid cells from epilepsy tissues, but two samples showed strong up-regulation of CCR2 in GBM (Figure 1c). Additionally, we detected CCR2 positive TAMs in three of six human GBM samples after immunoﬂuorescence staining while no CCR2 protein was expressed in IBA1+ cells of EP tissues (Figure 1d). This suggests the relevance of the CCR2/CCL2 signaling for myeloid cells in a subset of human GBMs. To ﬁnd a suitable model system, we investigated CCR2 expression in mice with BL6/J background. We observed CCR2 expression in TAMs of tumors of the syngeneic GL261 glioma mouse model both on RNA [25] and protein level (Figure 1e). Flow cytometric analyses revealed only minor expression of CCR2 on myeloid cells (CD11b+CD45+) in naïve mouse brains while CCR2 was up-regulated in tumor-bearing mice (Figure 1f). The percentage of CCR2+ cells within the CD11b+CD45+ myeloid cell population increased signiﬁcantly (Figure 1g) implicating their reactivity on the CCR2/CCL2 signaling pathway. Cancers 2020, 12, 1882 3 of 19 Cancers 2019, 11, x 3 of 2 Figure 1. CCR2 is expressed within human and murine glioma tissues. (a) Expression data of CCR2 from glioblastoma patients were used from TCGA Affymetrix U133A data (528 GBM patient samples). The relative gene expression of each patient is depicted (each column represents one patient). Black lines define the z-score threshold (0.5). (b) Human epilepsy (EP) and GBM brain tissue sections stained for CCR2. Scale bars 100 µm (n = 3–6). (c) Myeloid cells (CD11b+) were isolated from freshly homogenized human brain tissues, and RNA was extracted. Realtime-PCR for CCR2 is Figure 1. CCR2 is expressed within human and murine glioma tissues. 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu * p < 0.001 (unpaired Student’s t-test). Altogether, we found a remarkable expression of CCR2 in tumor-associated macrophages in human and murine glioblastomas. To understand the role of CCR2 within the tumor microenvironment, we used a suitable rodent tumor model in combination with Ccr2-/- mice. Altogether, we found a remarkable expression of CCR2 in tumor-associated macrophages in human and murine glioblastomas. To understand the role of CCR2 within the tumor microenvironment, we used a suitable rodent tumor model in combination with Ccr2-/- mice. 2.2. Diminished Inﬁltration of TAMs in Brain Tumor Tissues of Ccr2KO Mice 2.2. Diminished Inﬁltration of TAMs in Brain Tumor Tissues of Ccr2KO Mice 2.2. Diminished Inﬁltration of TAMs in Brain Tumor Tissues of Ccr2KO Mice TAMs were isolated from naïve and tumor-bearing wildtype (WT) as well as Ccr2-/- (Ccr2KO) mice. RNA has been extracted with subsequent gene expression analyses. As expected, Ccr2KO mice did not express Ccr2 following 21-days of glioma growth, in strong contrast with WT animals (Figure 2a). We sought to investigate homogenized brain tumor tissues for immune cell inﬁltration and observed a remarkable shift (Figure 2b). In Ccr2KO mice, the myeloid cell fraction (CD11b+CD45+) was signiﬁcantly reduced while the lymphocyte population (CD11b-CD45+) increased (Figure 2c). However, total numbers of T cells, whether eﬀector T cells or immune-suppressive regulatory T cells, were unchanged (Figure S1a–c), and B cell count was slightly reduced (Figure S1d). The decrease in TAMs was veriﬁed by immunoﬂuorescence staining (Figure 2d). In particular, intratumoral areas showed reduced numbers of TAMs in Ccr2-deﬁcient mice (Figure 2e) while peritumoral regions were not aﬀected (Figure 2f). We suggest that the altered ratio of myeloid cells and lymphocytes was likely based on diminished TAM inﬂux. To exclude basal diﬀerences in myeloid cell distribution in the CNS, microglia of naïve mice were counted revealing similar numbers and comparable morphology in wildtype and transgenic mice (Figure S2a,b). In gliomas, both microglia and macrophages can inﬁltrate the tumor tissue while their individual contribution is intensely debated [7,26]. Even if we did not analyze them separately in vivo, we stimulated microglia from naïve brain tissues and also generated macrophages from bone marrow with tumor-conditioned media. We observed that stimulated microglia and macrophages of both mouse strains expressed equal markers in vitro, e.g., IBA1, CD11b, CD68, and F4/80, respectively (Figure S2c–e) demonstrating viable and inducible myeloid cells independently of Ccr2. 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu (a) Expression data of CCR2 from glioblastoma patients were used from TCGA Aﬀymetrix U133A data (528 GBM patient samples). The relative gene expression of each patient is depicted (each column represents one patient). Black lines deﬁne the z-score threshold (0.5). (b) Human epilepsy (EP) and GBM brain tissue sections stained for CCR2. Scale bars 100 µm (n = 3–6). (c) Myeloid cells (CD11b+) were isolated from freshly homogenized 3 of 19 Cancers 2020, 12, 1882 Figure 1. CCR2 is expressed within human and murine glioma tissues. (a) Expression data of CCR2 from glioblastoma patients were used from TCGA Affymetrix U133A data (528 GBM patient samples). The relative gene expression of each patient is depicted (each column represents one patient). Black lines define the z-score threshold (0.5). (b) Human epilepsy (EP) and GBM brain tissue sections stained for CCR2. Scale bars 100 µm (n = 3–6). (c) Myeloid cells (CD11b+) were isolated from f hl h i d h b i ti d RNA t t d R lti PCR f CCR2 i Figure 1. CCR2 is expressed within human and murine glioma tissues. (a) Expression data of CCR2 from glioblastoma patients were used from TCGA Aﬀymetrix U133A data (528 GBM patient samples). The relative gene expression of each patient is depicted (each column represents one patient). Black lines deﬁne the z-score threshold (0.5). (b) Human epilepsy (EP) and GBM brain tissue sections stained for CCR2. Scale bars 100 µm (n = 3–6). (c) Myeloid cells (CD11b+) were isolated from freshly homogenized 4 of 19 Cancers 2020, 12, 1882 human brain tissues, and RNA was extracted. Realtime-PCR for CCR2 is presented (n = 6–15). (d) Representative images of frozen human brain sections stained for IBA1 and CCR2 are depicted (arrowheads deﬁne IBA1+ cells). Scale bars 100 µm (n = 3–6). (e) Murine brain tumor sections were analyzed for IBA1 and CCR2 on day 21 of glioma growth. A representative image of the intratumoral area is shown (square illustrates magniﬁcation of tumor tissue area; arrowheads deﬁne IBA1+ cells expressing CCR2). Scale bars 100 µm (n = 3). (f) Murine brain cells were stained with CD11b, CD45, and CCR2 antibodies and analyzed via ﬂow cytometry. Dot plots represent the expression of CCR2 of CD11b+CD45+ myeloid cells within naïve (N) and tumor-bearing (T) brains. (g) Graph depicts percentage of CCR2+ cells within the CD11b+CD45+ cell fraction (n = 10–12). 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu Ccr2-deﬁciency is known to aﬀect the migration of TAMs [23], however, their proliferative and apoptotic activity may be additionally inﬂuenced, which could explain diﬀerences in TAM counts of the intratumoral area. We, therefore, stained for Ki67 (Figure 2g) and DNA fragmentation (Figure 2i) of IBA1+ cells but could not detect changes in proliferation (Figure 2h) or apoptosis (Figure 2j). In line with the current literature, we assume that the reduced accumulation of TAMs in glioma tissue of Ccr2KO mice is predominantly due to diminished CCR2-dependent migration rather than alteration of their cellular activity. However, 70% of myeloid cells inﬁltrate the tumor tissue independently of the CCR2/CCL2 signaling axis (Figure 2e). 5 of 19 rs 2020, 12, 1882 5 o rs 2019, 11, x 5 o Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deficient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by flow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunofluorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines define the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunofluorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deﬁcient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). ** p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by ﬂow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by flow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunofluorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines define the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunofluorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the tumor area of WT and Ccr2KO mice (d21). Squares illustrate magnified areas. Scale bars 100 µm. (h,j) Graphs depict rate of proliferation (h) and calculation of apoptotic IBA1+ cells (j) (n = 6). p < 0.01; ns, Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deﬁcient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by ﬂow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunoﬂuorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines deﬁne the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunoﬂuorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the tumor area of WT and Ccr2KO mice (d21). Squares illustrate magniﬁed areas. Scale bars 100 µm. (h,j) Graphs depict rate of proliferation (h) and calculation of apoptotic IBA1+ cells (j) (n = 6). p < 0.01; ns, not signiﬁcant (unpaired Student’s t-test). 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deficient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by flow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunofluorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines define the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunofluorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the tumor area of WT and Ccr2KO mice (d21). Squares illustrate magnified areas. Scale bars 100 µm. (h,j) Graphs depict rate of proliferation (h) and calculation of apoptotic IBA1+ cells (j) (n = 6). p < 0.01; ns, i ifi ( i d S d ’ ) Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deﬁcient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by ﬂow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunoﬂuorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines deﬁne the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11). (d) Tumors of WT and Ccr2KO brains were analyzed via immunoﬂuorescence staining for myeloid cells (IBA1+; d21). Representative images of the tumor border are depicted. Dashed lines deﬁne the peritumoral area (pt; 100 µm region from tumor border into normal brain tissues) and surround the intratumoral area (it). Scale bars 100 µm. (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunoﬂuorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the tumor area of WT and Ccr2KO mice (d21) Squares illustrate Cancers 2020, 12, 1882 Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deficient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). ** p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by flow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10–11) (d) Tumors of WT and Ccr2KO brains were analyzed via Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deﬁcient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). ** p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni’s multiple comparison test). (b) Tumor-bearing brains of WT and Ccr2KO mice (d21) were analyzed by ﬂow cytometry. Representative dot plots show gates for TAMs (CD11b+CD45+) and lymphocytes (CC11b-CD45+). (c) The graph depicts the calculation of myeloid cells and lymphocytes (n = 10 11) (d) Tumors of WT and Ccr2KO brains were analyzed via immunoﬂuorescence staining for Figure 2. IBA1+ cells accumulate less in tumor-bearing Ccr2-deficient mouse brains. (a) RT-PCR for Ccr2 of CD11b+ cells from naïve (N) and tumor-bearing (T; d21) brain tissues of the wildtype (WT), as well as Ccr2KO (KO) mice, is presented (n = 3–4). ** p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni’s multiple comparison test). 2.1. CCR2-Expression of Microglia/Macrophages in Human and Murine Glioblastoma Tissu (e,f) The calculation for the number of IBA1+ cells within the it (e) and pt (f) area are illustrated (n = 6). (g,i) Representative images of immunoﬂuorescence staining for IBA1 and Ki67 (g) or apoptosis (i) in the tumor area of WT and Ccr2KO mice (d21). Squares illustrate magniﬁed areas. Scale bars 100 µm. (h,j) Graphs depict rate of proliferation (h) and calculation of apoptotic IBA1+ cells (j) (n = 6). p < 0.01; ns, not signiﬁcant (unpaired Student’s t-test). Cancers 2020, 12, 1882 6 of 19 2.3. Ccr2-Deﬁciency Leads to Accelerated Brain Tumor Growth 2.3. Ccr2-Deﬁciency Leads to Accelerated Brain Tumor Growth In other tumor models, it has been described that Ccr2-deﬁciency led to reduced tumor sizes [20]. Hence, we analyzed glioma progression at consecutive time points using MRI (Figure 3a). Unexpectedly, we measured an increase of tumor volumes at day 7 and 14, while sizes even doubled in Ccr2KO mice compared to WT at day 21 (Figure 3b). Early animal loss in the Ccr2KO group up to day 21 has not been observed. However, survival experiments were not performed. Despite augmented tumor progression, no diﬀerences in glioma cell invasion were noticed after the histopathological examination (Figure 3c). Moreover, we assessed the cellular activity of tumor tissues and discovered no diﬀerences in apoptosis (Figure 3d,e) but an increased cell proliferation in Ccr2KO animals (Figure 3f) by up to 25% (Figure 3g). The rapid and uncontrolled tumor cell proliferation restricts the ability of oxygen. Therefore, hypoxia is a typical feature of gliomas [27,28]. Here, we analyzed hypoxia-regulated molecules such as HIF1α and BNIP3 that are commonly up-regulated during tumor progression [27–29]. HIF1α, as a pivotal hallmark in response to hypoxia, showed strong expression (Figure 3h), but no diﬀerences between mouse strains were observed (Figure 3i). In our glioma model, we found BNIP3 located in the nuclei (Figure S3a). This localization is associated with increased tumor cell survival [29,30]. But again, WT and Ccr2KO mice revealed comparable results (Figure S3b). Thus, despite the higher proliferative activity of tumors in Ccr2KO mice, we could not detect signiﬁcant changes in hypoxic area distribution implicating a suﬃcient supply of oxygen and nutrients in these tumors. In addition, we investigated the expression of immune relevant molecules that can modulate the tumor microenvironment. We focused on interferon-gamma (IFNγ), a cytokine which facilitates antitumor immunity [31] and programmed death ligand 1 (PDL1) relevant for mediation of immunosuppression [32]. Interestingly, we found reduced IFNγ expression (Figure 3j,k) and increased PDL1 signal (Figure 3l,m) in gliomas of Ccr2-/- mice that indicated a rather immunosuppressive milieu in these tumors. Our results demonstrate that the reduced number of inﬁltrating TAMs is accompanied by accelerated glioma growth, an increased tumor cell proliferation, and a more tumor supportive micromilieu in Ccr2KO mice. 2.4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Genes .4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Ge Despite enormous research eﬀorts to understand the TAM function in glioma, their precise role remains unclear. Recent studies identiﬁed these cells as a mixed population with pro-inﬂammatory and immune suppressive properties [14,33], but ultimately promote tumor growth [25]. Consequently, TAMs were likely involved in the tumor progression of recently applied glioma models. To assess the general functionality of TAMs, we validated class-identifying molecules. Characteristic surface markers of tumor-associated macrophages, like CD11b and CD68, were found in gliomas of both mouse strains, without noticeable diﬀerences in expression (Figure 4a,b). Furthermore, we investigated antigen-presenting molecules (MHCI, MHCII) as well as costimulatory ligands (CD80, CD86) important for the induction of immune cell responses. As previously deﬁned, only low amounts of TAMs carry MHCI in murine glioma tissues [34] which we were also able to detect in Ccr2KO mice (Figure 4c). The expression of MHCII and co-stimulatory molecules of Ccr2KO were comparable to WT (Figure 4d,e). However, analyzing the expression of various immune-regulatory genes, belonging to the family of transcription factors or cytokines, revealed changes of TAMs in glioma-bearing hemispheres of Ccr2KO mice (Figure 4f) while the basic expression of some genes of microglia from naïve mice already vary (Table S2). We identiﬁed Stat3, Irf7, Cox2, and Ifnβ as being up-regulated, while Il1β and Il12α show down-regulation under tumor condition. Ifnβ, that displayed strong up-regulation, has previously been described to induce expression of PDL1 in a subpopulation of myeloid cells, the myeloid-derived suppressor cells (MDSCs) [35] which are known to have immune-suppressive functions. Thus, we investigated the expression of PDL1 by IBA1+ cells within the tumor area. We found a slight increase of TAMs that express PDL1 but without reaching signiﬁcance (Figure S4). 7 of 19 Cancers 2020, 12, 1882 Cancers 2019, 11, x 7 of 20 Figure 3. Glioma growth is enhanced in Ccr2-deficient mice. (a) Tumor sizes of gliomas were measured with MRI on day 7, 14, and 21 after inoculation with GL261 cells. Representative MRI images are shown for the WT (upper row) and Ccr2KO (lower row) mice. Red dashed lines define tumor borders. (b) Graph displays calculated tumor volumes (n = 10–12). * p < 0.001 (one-way ANOVA and Bonferroni´s multiple comparison test). (c) Representative images of hematoxylin-eosin- stained tissue sections of tumor-bearing WT and Ccr2KO brains are shown for day 21 (n = 4–5). Squares indicate magnified regions of the tumor border. Scale bars 1000 µm. .4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Ge (d,f) Apoptosis (d) and proliferation (f) of the intratumoral area (d21) were analyzed by immunofluorescence staining of ApopTag (d) and Ki67 (f). Scale bars 100 µm. (e,g) Graphs depict the amount of apoptotic (e) and proliferative (g) cells within glioma tissues (n = 6). (h) HIF1α was stained to detect hypoxia in tumor tissues (d21). Squares indicate magnified regions. Scale bars 100 µm. (i) Calculation for area of HIF1α expression (n = 5). (j–m) Tissues were stained for IFNγ (j) and PDL1 (l). Scale bars 100 µm. Positive areas of IFNγ (k) and PDL1 were calculated (m) (n = 4–5). *** p < 0.001; * p < 0.05; ns, not significant (unpaired Student’s t-test). Figure 3. Glioma growth is enhanced in Ccr2-deﬁcient mice. (a) Tumor sizes of gliomas were measured with MRI on day 7, 14, and 21 after inoculation with GL261 cells. Representative MRI images are shown for the WT (upper row) and Ccr2KO (lower row) mice. Red dashed lines deﬁne tumor borders. (b) Graph displays calculated tumor volumes (n = 10–12). * p < 0.001 (one-way ANOVA and Bonferroni’s multiple comparison test). (c) Representative images of hematoxylin-eosin-stained tissue sections of tumor-bearing WT and Ccr2KO brains are shown for day 21 (n = 4–5). Squares indicate magniﬁed regions of the tumor border. Scale bars 1000 µm. (d,f) Apoptosis (d) and proliferation (f) of the intratumoral area (d21) were analyzed by immunoﬂuorescence staining of ApopTag (d) and Ki67 (f). Scale bars 100 µm. (e,g) Graphs depict the amount of apoptotic (e) and proliferative (g) cells within glioma tissues (n = 6). (h) HIF1α was stained to detect hypoxia in tumor tissues (d21). Squares indicate magniﬁed regions. Scale bars 100 µm. (i) Calculation for area of HIF1α expression (n = 5). (j–m) Tissues were stained for IFNγ (j) and PDL1 (l). Scale bars 100 µm. Positive areas of IFNγ (k) and PDL1 were calculated (m) (n = 4–5). * p < 0.001; * p < 0.05; ns, not signiﬁcant (unpaired Student’s t-test). Figure 3. Glioma growth is enhanced in Ccr2-deficient mice. (a) Tumor sizes of gliomas were measured with MRI on day 7, 14, and 21 after inoculation with GL261 cells. Representative MRI images are shown for the WT (upper row) and Ccr2KO (lower row) mice. Red dashed lines define tumor borders. (b) Graph displays calculated tumor volumes (n = 10–12). .4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Ge 8 of 19 8 of 19 Cancers 2020, 12, 1882 Cancers 2020, 12, 1882 8 of 19 Cancers 2019, 11, x 9 of 20 Figure 4. Tumor-associated macrophages (TAMs) of WT and Ccr2KO mice carry similar surface markers but show differences in expression of transcription factors as well as cytokines at day 21 of tumor growth. (a–c) Representative images of brain tumor sections of WT and Ccr2KO mice stained for IBA1 and CD11b (a) or CD68 (b) or MHCI (c) are depicted (n = 4–5). Squares indicate magnified regions (a,b). Arrowheads define IBA1+ cells expressing MHCI (c). Scale bars 100 µm. (d) Brain tumor suspensions were analyzed by flow cytometry after staining with CD11b, CD45, and MHCII, CD80, or CD86 antibodies. Histograms represent molecule expression of CD11b+CD45+ myeloid cells within WT and Ccr2KO mice including isotype controls. (e) The graph depicts the percentage of molecule expression (n = 6–14). Blue lines define basic molecule expression in naïve mice. (f) RT-PCRs for Figure 4. Tumor-associated macrophages (TAMs) of WT and Ccr2KO mice carry similar surface markers but show diﬀerences in expression of transcription factors as well as cytokines at day 21 of tumor growth. (a–c) Representative images of brain tumor sections of WT and Ccr2KO mice stained for IBA1 and CD11b (a) or CD68 (b) or MHCI (c) are depicted (n = 4–5). Squares indicate magniﬁed regions (a,b). Arrowheads deﬁne IBA1+ cells expressing MHCI (c). Scale bars 100 µm. (d) Brain tumor suspensions were analyzed by ﬂow cytometry after staining with CD11b, CD45, and MHCII, CD80, or CD86 antibodies. Histograms represent molecule expression of CD11b+CD45+ myeloid cells within WT and Ccr2KO mice including isotype controls. (e) The graph depicts the percentage of molecule expression (n = 6–14) Blue lines deﬁne basic molecule expression in naïve mice (f) RT-PCRs Figure 4. Tumor-associated macrophages (TAMs) of WT and Ccr2KO mice carry similar surface markers but show differences in expression of transcription factors as well as cytokines at day 21 of tumor growth. (a–c) Representative images of brain tumor sections of WT and Ccr2KO mice stained for IBA1 and CD11b (a) or CD68 (b) or MHCI (c) are depicted (n = 4–5). Squares indicate magnified regions (a,b). Arrowheads define IBA1+ cells expressing MHCI (c). Scale bars 100 µm. (d) Brain tumor suspensions were analyzed by flow cytometry after staining with CD11b, CD45, and MHCII, CD80, or CD86 antibodies. .4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Ge * p < 0.001 (one-way ANOVA and Bonferroni´s multiple comparison test). (c) Representative images of hematoxylin-eosin- stained tissue sections of tumor-bearing WT and Ccr2KO brains are shown for day 21 (n = 4–5). Squares indicate magnified regions of the tumor border. Scale bars 1000 µm. (d,f) Apoptosis (d) and proliferation (f) of the intratumoral area (d21) were analyzed by immunofluorescence staining of ApopTag (d) and Ki67 (f). Scale bars 100 µm. (e,g) Graphs depict the amount of apoptotic (e) and proliferative (g) cells within glioma tissues (n = 6). (h) HIF1α was stained to detect hypoxia in tumor tissues (d21). Squares indicate magnified regions. Scale bars 100 µm. (i) Calculation for area of HIF1α expression (n = 5). (j–m) Tissues were stained for IFNγ (j) and PDL1 (l). Scale bars 100 µm. Positive areas of IFNγ (k) and PDL1 were calculated (m) (n = 4–5). * p < 0.001; * p < 0.05; ns, not significant (unpaired Student’s t test) Figure 3. Glioma growth is enhanced in Ccr2-deﬁcient mice. (a) Tumor sizes of gliomas were measured with MRI on day 7, 14, and 21 after inoculation with GL261 cells. Representative MRI images are shown for the WT (upper row) and Ccr2KO (lower row) mice. Red dashed lines deﬁne tumor borders. (b) Graph displays calculated tumor volumes (n = 10–12). * p < 0.001 (one-way ANOVA and Bonferroni’s multiple comparison test). (c) Representative images of hematoxylin-eosin-stained tissue sections of tumor-bearing WT and Ccr2KO brains are shown for day 21 (n = 4–5). Squares indicate magniﬁed regions of the tumor border. Scale bars 1000 µm. (d,f) Apoptosis (d) and proliferation (f) of the intratumoral area (d21) were analyzed by immunoﬂuorescence staining of ApopTag (d) and Ki67 (f). Scale bars 100 µm. (e,g) Graphs depict the amount of apoptotic (e) and proliferative (g) cells within glioma tissues (n = 6). (h) HIF1α was stained to detect hypoxia in tumor tissues (d21). Squares indicate magniﬁed regions. Scale bars 100 µm. (i) Calculation for area of HIF1α expression (n = 5). (j–m) Tissues were stained for IFNγ (j) and PDL1 (l). Scale bars 100 µm. Positive areas of IFNγ (k) and PDL1 were calculated (m) (n = 4–5). * p < 0.001; * p < 0.05; ns, not signiﬁcant (unpaired Student’s t-test). 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice Sustained angiogenesis is regarded as a central hallmark of cancer development, and stimulation of blood vessel growth leads to the mandatory supply of nutrients and oxygen during glioma growth [16]. Changes in vascularization may be responsible for increased tumor sizes. Therefore, we analyzed the vascular structure by immunoﬂuorescence staining. To exclude basal diﬀerences, the contralateral hemisphere of WT and Ccr2KO brains were investigated (Figure S5a), revealing comparable vessel densities (Figure S5b) and vascularized areas (Figure S3c). Comparing tumor tissue of both mouse strains (Figure 5a), we found similar vessel counts (Figure 5b) and no changes in the blood vessel covered areas (Figure 5c). Additionally, the vasculature of the peritumoral area appeared to be unaﬀected by Ccr2-deﬁciency (Figure S5d–f). Overall, a similar vascular structure of both mouse strains has been characterized. Sustained angiogenesis is regarded as a central hallmark of cancer development, and stimulation of blood vessel growth leads to the mandatory supply of nutrients and oxygen during glioma growth [16]. Changes in vascularization may be responsible for increased tumor sizes. Therefore, we analyzed the vascular structure by immunofluorescence staining. To exclude basal differences, the contralateral hemisphere of WT and Ccr2KO brains were investigated (Figure S5a), revealing comparable vessel densities (Figure S5b) and vascularized areas (Figure S3c). Comparing tumor tissue of both mouse strains (Figure 5a), we found similar vessel counts (Figure 5b) and no changes in the blood vessel covered areas (Figure 5c). Additionally, the vasculature of the peritumoral area appeared to be unaffected by Ccr2-deficiency (Figure S5d–f). Overall, a similar vascular structure of both mouse strains has been characterized. Figure 5. Glioma vasculature is unaffected by Ccr2-deficiency. (a) Representative images of brain tumor sections of WT and Ccr2KO mice stained for CD31 are depicted (d21). Scale bars 100 µm. (b,c) Graphs indicate vessel count (b) and vascularized area (c) within the tumor tissues (n = 6). (d) Confocal microscopy of brain tumor sections from mice perfused with FITC-Lectin and stained for CD31 (overview, mosaic). Square indicates the magnified region (63×). T, tumor tissue; dashed line, tumor border. (e) The graph presents the percentage of perfused vessels (n = 3–4). ns, not significant (unpaired Student’s t-test). Figure 5. Glioma vasculature is unaﬀected by Ccr2-deﬁciency. (a) Representative images of brain tumor sections of WT and Ccr2KO mice stained for CD31 are depicted (d21). Scale bars 100 µm. .4. TAMs of Ccr2KO Mice Show Classical Markers but Exhibit a Modiﬁed Expression of Inﬂammatory Ge Histograms represent molecule expression of CD11b+CD45+ myeloid cells within WT and Ccr2KO mice including isotype controls. (e) The graph depicts the percentage of molecule expression (n = 6–14). Blue lines define basic molecule expression in naïve mice. (f) RT-PCRs for indicated genes of CD11b+ cells from tumor-bearing brain tissues of WT as well as Ccr2KO mice are Figure 4. Tumor-associated macrophages (TAMs) of WT and Ccr2KO mice carry similar surface markers but show diﬀerences in expression of transcription factors as well as cytokines at day 21 of tumor growth. (a–c) Representative images of brain tumor sections of WT and Ccr2KO mice stained for IBA1 and CD11b (a) or CD68 (b) or MHCI (c) are depicted (n = 4–5). Squares indicate magniﬁed regions (a,b). Arrowheads deﬁne IBA1+ cells expressing MHCI (c). Scale bars 100 µm. (d) Brain tumor suspensions were analyzed by ﬂow cytometry after staining with CD11b, CD45, and MHCII, CD80, or CD86 antibodies. Histograms represent molecule expression of CD11b+CD45+ myeloid cells within WT and Ccr2KO mice including isotype controls. (e) The graph depicts the percentage of molecule expression (n = 6–14). Blue lines deﬁne basic molecule expression in naïve mice. (f) RT-PCRs for indicated genes of CD11b+ cells from tumor-bearing brain tissues of WT as well as Ccr2KO mice are presented (n = 3–4). Blue line deﬁnes basic molecule expression in naïve WT mice. *** p < 0.001; * p < 0.05 (unpaired Student’s t-test). Cancers 2020, 12, 1882 9 of 19 Thus, myeloid cells accumulating in intratumoral areas of Ccr2KO mice are characterized by similar expression of TAM-markers and surface molecules required for T cell activation. However, transcription factors as well as soluble molecules were expressed diﬀerently without meaningful polarization of the myeloid cells in Ccr2-deﬁcient mice. Cancers 2019, 11, x 10 of 20 presented (n = 3–4). Blue line defines basic molecule expression in naïve WT mice. * p < 0.001; p < 0.01; * p < 0.05 (unpaired Student’s t-test). 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice In addition, vessels of tumors in Ccr2KO mice showed a greater coverage with pericytes (Figure 6d,e), which can also be a sign for higher maturity of blood vessels [37]. We recently demonstrated that TAMs are often associated with glioma vasculature and involved in angiogenesis by expression of pro-angiogenic factors [25]. Interestingly, despite the reduction of TAM counts in transgenic mice, the interaction of IBA1+ cells with tumor blood vessels remained at approximately 18% (Figure 6f,g). This indicates a preference of TAMs for the perivascular niche in Ccr2KO mice, which could facilitate the improved vessel functionality. Therefore, we investigated whether TAMs of Ccr2KO mice are involved in pericyte recruitment or angiogenesis by expression of various genes such as Pdgfβ, Ang1/2, and Mmp2/9, but were not able to discover signiﬁcant alterations compared to the wildtype (Figure S5g). However, Ccr2-deﬁciency caused a reduction of the angiogenic molecule Vegf in TAMs of the tumor-bearing hemisphere (Figure 6h), likely due to improved nutrient and oxygen supply of stabilized vessels. Cancers 2019, 11, x 11 of 20 dependent on Ccr2-deficiency. In addition, vessels of tumors in Ccr2KO mice showed a greater coverage with pericytes (Figure 6d,e), which can also be a sign for higher maturity of blood vessels [37]. We recently demonstrated that TAMs are often associated with glioma vasculature and involved in angiogenesis by expression of pro-angiogenic factors [25]. Interestingly, despite the reduction of TAM counts in transgenic mice, the interaction of IBA1+ cells with tumor blood vessels remained at approximately 18% (Figure 6f,g). This indicates a preference of TAMs for the perivascular niche in Ccr2KO mice, which could facilitate the improved vessel functionality. Therefore, we investigated whether TAMs of Ccr2KO mice are involved in pericyte recruitment or angiogenesis by expression of various genes such as Pdgfβ, Ang1/2, and Mmp2/9, but were not able to discover significant alterations compared to the wildtype (Figure S5g). However, Ccr2-deficiency caused a reduction of the angiogenic molecule Vegf in TAMs of the tumor-bearing hemisphere (Figure 6h), likely due to improved nutrient and oxygen supply of stabilized vessels. Thus, we observed enhanced vascular integrity in tumors of Ccr2KO mice, which may be one of the responsible factors for augmented tumor growth. Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deficient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice (b,c) Graphs indicate vessel count (b) and vascularized area (c) within the tumor tissues (n = 6). (d) Confocal microscopy of brain tumor sections from mice perfused with FITC-Lectin and stained for CD31 (overview, mosaic). Square indicates the magniﬁed region (63×). T, tumor tissue; dashed line, tumor border. (e) The graph presents the percentage of perfused vessels (n = 3–4). ns, not signiﬁcant (unpaired Student’s t-test). Figure 5. Glioma vasculature is unaffected by Ccr2-deficiency. (a) Representative images of brain tumor sections of WT and Ccr2KO mice stained for CD31 are depicted (d21). Scale bars 100 µm. (b,c) Graphs indicate vessel count (b) and vascularized area (c) within the tumor tissues (n = 6). (d) Confocal microscopy of brain tumor sections from mice perfused with FITC-Lectin and stained for CD31 (overview, mosaic). Square indicates the magnified region (63×). T, tumor tissue; dashed line, tumor border. (e) The graph presents the percentage of perfused vessels (n = 3–4). ns, not significant (unpaired Student’s t-test). Figure 5. Glioma vasculature is unaﬀected by Ccr2-deﬁciency. (a) Representative images of brain tumor sections of WT and Ccr2KO mice stained for CD31 are depicted (d21). Scale bars 100 µm. (b,c) Graphs indicate vessel count (b) and vascularized area (c) within the tumor tissues (n = 6). (d) Confocal microscopy of brain tumor sections from mice perfused with FITC-Lectin and stained for CD31 (overview, mosaic). Square indicates the magniﬁed region (63×). T, tumor tissue; dashed line, tumor border. (e) The graph presents the percentage of perfused vessels (n = 3–4). ns, not signiﬁcant (unpaired Student’s t-test). Still, tumor blood vessels can be characterized by increased permeability and leakiness, leading to impaired function [36]. To study the integrity of blood vessels in tumor mice we analyzed vessel perfusion and stability. Injection of FITC-labeled lectin (Figure 5d) revealed comparable fractions of Still, tumor blood vessels can be characterized by increased permeability and leakiness, leading to impaired function [36]. To study the integrity of blood vessels in tumor mice we analyzed vessel perfusion and stability. Injection of FITC-labeled lectin (Figure 5d) revealed comparable fractions 10 of 19 Cancers 2020, 12, 1882 of lectin perfused tumor vessels in WT and Ccr2KO mice (Figure 5e). Interestingly, the albumin covered area was reduced in glioma tissues of Ccr2KO animals (Figure 6a,b) accompanied by less albumin-stained tumor vessels (Figure 6c) indicating increased stability and less permeability of blood vessels dependent on Ccr2-deﬁciency. 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not significant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni´s multiple comparison test). Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deﬁcient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not signiﬁcant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni´s multiple comparison test). Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deficient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not significant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni´s multiple comparison test). Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deﬁcient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not signiﬁcant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni´s multiple comparison test). Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deficient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not significant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not significant (one-way ANOVA and Bonferroni´s multiple comparison test). Figure 6. Vascular integrity is improved of glioma grown in Ccr2-deﬁcient mice. (a,d,f) Representative images of brain tumor sections of WT and Ccr2KO mice (d21) stained for CD31 and albumin (a), Desmin (d) or IBA1 (f) are depicted. Arrowheads indicate vessels in contact with IBA1+ cells (f). Scale bars 100 µm. (b,c,e,g) Graphs present area of albumin staining (b), vessels colocalized with albumin (c), pericyte-covered vessels (e) and vessels interacting with IBA1+ cells (g). * p < 0.05; ns, not signiﬁcant (unpaired Student’s t-test). (h) RT-PCR for Vegf of CD11b+ cells (myeloid cells) from naïve (N) and tumor-bearing (T) brain tissues of WT as well as Ccr2KO mice is depicted (d21; n = 3–4). ** p < 0.01; * p < 0.05; ns, not signiﬁcant (one-way ANOVA and Bonferroni´s multiple comparison test). 3. Discussion Overall, we investigated the impact of Ccr2-deficiency in the glioma microenvironment on Thus, we observed enhanced vascular integrity in tumors of Ccr2KO mice, which may be one of the responsible factors for augmented tumor growth. 3. 3. Discussion Overall, we investigated the impact of Ccr2-deﬁciency in the glioma microenvironment on tumor progression. We have been able to show variable CCR2 expression in human GBM tissues and high expression in murine gliomas. Using the syngeneic GL261 glioma model, increased tumor volumes in Ccr2-/- mice were measured, accompanied by reduced inﬁltration of TAMs and higher integrity of blood vessels. The decrease in myeloid cell recruitment has previously been observed in Ccr2-knockout tumor models [18,20]. However, analyzing the impact of Ccr2-deﬁciency on tumor sizes and vascularization led to diﬀerent results. In prostate cancer, tumor volumes were unaﬀected, while cell proliferation and angiogenesis were diminished [18]. In lung carcinoma, smaller tumors with strong vascular remodeling were found [20]. Thus, the relevance of CCR2 on tumor progression varies and depends on the tumor’s environmental context and location. The glioma microenvironment in the immune-privileged brain is diﬀerent due to resident microglia that accumulate in addition to peripheral macrophages [5]. A recent study described only a small impact on the survival of glioma-bearing Ccr2 transgenic mice whereby tumor volumes were not evaluated [38]. Due to our experimental set-up with a focus on glioma biology and a deﬁned endpoint, we were not able to link our results to survival data of this study. Nevertheless, the authors observed reduced numbers of TAMs with immunosuppressive features as well [38]. Targeting the ligand of CCR2, CCL2, in prostate cancer [18,19] and glioma [17,21] led to a reduction of tumor growth and prolonged survival. Under these conditions, not only the recruitment of TAMs is inhibited but also the attraction of regulatory T cells via CCR4 [22] and the autocrine mechanism on glioma cells, expressing CCL2 and CCR2 [39], is interrupted [18,40]. Thus, observations in our used knockout mouse model could be referred to diminished TAM inﬁltration, while targeting CCL2 led to additional inhibitory eﬀects of the CCR2/CCL2 pathway. Following up on our experiments, Ccr2-knockout impaired the intratumoral TAM accumulation but was not entirely impeded. Still, 70% of IBA1+ cells were recruited into the glioma area independently of the CCR2/CCL2 signal. Considering current literature, it is assumed that only macrophages express CCR2 [21,22]. This would imply, preferential inﬁltration of microglia into gliomas of Ccr2-/- mice because these immune cells can migrate independently of CCR2. However, ﬁnal evidence for this hypothesis is lacking, and we were not diﬀerentiating between these both cell populations. 2.5. Improved Vascular Integrity in Ccr2-Deﬁcient Mice 2.5. Improved Vascular Integrity in Ccr2-Deficient Mice Discussion Overall, we investigated the impact of Ccr2-deficiency in the glioma microenvironment on Thus, we observed enhanced vascular integrity in tumors of Ccr2KO mice, which may be one of the responsible factors for augmented tumor growth. Cancers 2020, 12, 1882 11 of 19 11 of 19 3. Discussion Even if CCL2 is believed to be the major recruiter of TAMs in gliomas [15,16,41,42], other factors like CX3CL1 [43], CXCL12, M-CSF and GM-CSF [44] can mobilize TAMs into brain tumor areas, and become relevant under Ccr2-deﬁciency. Besides IBA1+ cells, we also investigated the inﬁltration of eﬀector T cells as well as regulatory T cells but detected no signiﬁcant diﬀerences while B cell counts were slightly reduced. A more diﬀerential classiﬁcation of T and B lymphocytes and their distribution was not performed, further challenging a detailed characterization of the immune cell composition in Ccr2-/- mice. Remaining intratumoral TAMs in Ccr2-/- mice appeared functional and showed classical features of the monocytic lineage, characterized by carrying IBA1, CD68, CD11b as well as antigen-presenting molecules. Isolated CD11b+ cells from tumors of both mouse strains expressed anti- and pro-inﬂammatory molecules, revealing a mixed phenotype as expected for the wildtype [33]. We referred to this cell population from tumor-bearing mice to myeloid cells. This cell population could be composed of microglia and macrophages which carry similar surface markers [26], while neutrophils could be precluded in this glioma model [34]. We decided not to further stratify subpopulations based on lacking evidence of their speciﬁc functions. It should be noted that we observed partly diﬀerent basal expression of genes in microglia of naïve wildtype and Ccr2-/- animals. We assume that the detected diﬀerences in gene expression of myeloid cells during glioma progression are relevant, but at this point, we are not able to pre-exclude an inﬂuence of divergent baseline values. The molecular signature of TAMs did not display a speciﬁc tumor-supportive polarization nor appear to induce the production of immunosuppressive cytokines Il10 or Tgfβ. Nevertheless, we detected a reduction of markers like iNos, Il1β, and Il12α, important for tumor growth inhibition [43,45], when comparing Ccr2-deﬁcient TAMs to wildtype cells. Furthermore, CD11b+ cells 12 of 19 Cancers 2020, 12, 1882 of Ccr2KO mice overexpressed Ifnβ and Stat3, molecules that are relevant for the immunosuppressive phenotype of MDSCs [35]. However, we could not ﬁnd elevated PDL1 expression within the TAMs of Ccr2-deﬁcient mice that should be induced by Ifnβ [35]. Even if PDL1 is unchanged in the IBA1 cell population, we observed an overall increase of PDL1 expression in the tumor tissue of Ccr2-/- mice. 3. Discussion Notably, tumor cells can utilize the PD1/PDL1 pathway to circumvent the immune surveillance by expression of PDL1 [32], and in human glioma, PDL1 was found to be a negative prognostic indicator [46]. While PDL1 is expressed by tumor cells and immune cells, IFNγ is primarily expressed by T cells as well as NK cells, and characterized as an anti-proliferative agent [31]. We did not co-stain IFNγ with immune cell markers. Nevertheless, we found reduced IFNγ positive staining in the glioma area. These data implicate an attenuated anti-tumor response and a rather immunosuppressive tumor micromilieu in Ccr2-deﬁcient mice which could support the enhanced glioma progression. Nutritional vessel supply and tumor-induced angiogenesis are important malignant characteristics of the GBM and are regarded as pivotal hallmarks of carcinogenesis [16]. Here, the blood-brain barrier integrity is frequently lost due to the angiogenic activity of the tumor [47], which allows for passive transport of cells and proteins as well as the reorganization of the vascular structure. We found higher pericyte coverage of tumor blood vessels and less albumin extravasation. Pericytes retain blood-brain barrier function and are known to be involved in vessel stability [37]. Our data implicate reduced blood-brain barrier degradation in Ccr2-deﬁcient mice, which has previously been demonstrated in other brain diseases [48]. We speculate that the tightened blood-brain barrier supports the decreased inﬁltration of myeloid cells from the periphery and encourages tumor maintenance. Our study aimed to elucidate the role of Ccr2-deﬁciency within the tumor microenvironment. Here, we focused on the myeloid cell population due to their well-characterized CCR2 expression [9,10], but also other leukocytes and endothelial cells express CCR2 under respective conditions [49,50]. Indeed, we found CCR2 positive cells negative for IBA1 in human and murine tumor tissues. We speculate that these are mainly glioma cells that are known to express CCR2 [39]. However, tumor microenvironmental cells other than the myeloid cells were not analyzed in detail. Thus, we cannot pre-exclude their contribution to the observed eﬀects in the glioma mouse model used. In contrast with previous studies, which targeted the CCR2/CCL2 pathway primarily in alternative experimental models [40,51], we found enhanced tumor volumes in Ccr2-/- mice. However, divergent data obtained in transgenic mouse models and under treatment conditions have been described previously [52]. It should be considered that targeting antibodies ubiquitously aﬀect all receptors or ligands, respectively. 3. Discussion In comparison, by using the model applied here, only cells of the tumor microenvironment (e.g., myeloid cells, endothelial cells) possess Ccr2-deﬁciency, while glioma cells still express Ccr2, hence retaining functionality towards CCL2 and other signals. We assume that if the entire CCR2/CCL2 signaling axis is inﬂicted by antibody treatment, tumor growth is inhibited, while speciﬁc knockout of tumor microenvironmental cells by using a Ccr2-/- transgenic mice leads to enhanced glioma progression. This progression is accompanied by reduced TAM accumulation, the strong proliferation of tumor cells, suﬃcient oxygen, and nutrient supply of the tumor tissue, ultimately supported by stabilized vasculature in Ccr2-/- mice. We speculate that CCR2-dependent myeloid cells are crucial to controlling glioma growth while the remaining CCR2-independent TAMs are unable to arrest or decelerate tumor expansion. We demonstrate that a more detailed investigation of TAMs is required to identify and characterize tumor-controlling subpopulations, which likely helps to ﬁnd more eﬀective treatment options. 4.4. MRI Analysis Animals were measured with a 7.0 Tesla animal scanner (Bruker Pharma Scan®, Billerica, MA, USA). To achieve high contrast and resolution, contrast agent Magnevist® (Bayer AG, Leverkusen, Germany) was injected into the tail vein. Mice were anesthetized with 1.5–2.0% isoﬂurane (Forene, Abbot, Abbot Park, IL, USA) in a narcotic mixture, together with O2/N2O (30/70%). Brain slices were recorded with T1- and T2-weighted 2-dimensional turbo spin-echo sequences by Paravision 4.0 software (Bruker Bio Spin®, Ettlingen, Germany). Volumes of tumors were calculated with Analyze 5.0 (AnalyzeDirect, Overland Park, KS, USA). 4.3. Intracerebral Tumor Cell Inoculation The tumor cell line GL261 was cultivated in high-glucose Dulbecco´s modiﬁed Eagles medium (DMEM) Gibco™(ThermoFisher Scientiﬁc, Waltham, MA, USA) and contained stable glutamine and sodium pyruvate, 10% fetal calf serum, 100 units of penicillin/mL and 100 µg/mL of streptomycin. Tumor cells were incubated for three days at 37 ◦C and 5% CO2 atmosphere until they reached a conﬂuence of 80%. Cells were diluted in PBS. 2 × 104 cells/µL were used for stereotactic cell inoculation. Anesthetized mice were ﬁxed in a stereotactic frame. The position for stereotactic implantation was 1 mm anterior and 2 mm lateral to the right from bregma. Via Hamilton syringe (Roth, Karlsruhe, Germany) 1 µL cell solution was injected into parenchyma at 3 mm depth. 4.1. Human Specimens Tissue samples were obtained during therapeutic surgical treatment (Department of Neurosurgery, Charité Universitätsmedizin Berlin, Germany) from 2013 to 2014. Here, epilepsy patients (6 cases) Cancers 2020, 12, 1882 13 of 19 who underwent temporal pole resection, and patients suﬀered from GBM (15 cases) were included. Neuropathologists assessed WHO tumor grade by standard prognostic markers. who underwent temporal pole resection, and patients suﬀered from GBM (15 cases) were included. Neuropathologists assessed WHO tumor grade by standard prognostic markers. All procedures involving human specimens were carried out following the ethical standards of the national research committee and consistent with the Helsinki Declaration and its later amendments or comparable ethical standards. Approval of the Ethical Committee of Charité-Universitätsmedizin Berlin was received (application number: EA4/065/13) and all analyses were carried out following the deﬁned obligations of scientiﬁc working with patient material. Informed consent was obtained from all subjects. j Tissue samples were embedded in 4% PFA for 24 h and subsequently dehydrated in a serial dilution of sucrose. Afterward, samples were frozen in liquid nitrogen. Frozen sections of 10 µm were prepared. Another tissue part was used for microglia/macrophage isolation. Gene expression data were achieved from the GBM patient dataset available through The Cancer Genome Atlas (TCGA; Aﬀymetrix U133A [53,54]). 4.2. Animals C57Bl6/Jmice(WT)wereobtainedfromCharlesRiverLaboratories(Sulzfeld, Germany). Homozygous Ccr2-/- animals (C57Bl6/J background; [23,55]) were received from Prof. Dr. Mathias Heikenwälder (HMGU Munich and DKFZ Heidelberg, Germany) and bred at FEM Bayer. Experiments were performed with sex- and age-matched animals. All animal experiments were conducted according to German Laws for Animal Protection and the National Institute of Health Guidelines for Care and Use of Laboratory Animals (LaGeSo No. G0152/09; G0281/14). 4.8. Immunoﬂuorescence Staining For each tumor, 3 to 8 sections were investigated and up to 9 images per section were taken depending on tumor size and staining. Images were analyzed by ImageJ Software (NIH; https://imagej.nih.gov/ij/). 4.8. Immunoﬂuorescence Staining Brain sections were blocked with PBS/0.5% Casein (Sigma-Aldrich) for 30 min. The following primary antibodies were used: rb anti-IBA1 (1:250; Wako Chemicals, Neuss, Germany), gt anti-IBA1 (1:100; Abcam, Cambridge, UK), rat anti-CD31 (1:50; BD Pharmingen, Heidelberg, Germany), rat anti-CD4 (1:50; BD Pharmingen), rat anti-CD8a (1:50; BD Pharmingen), rat anti-CD45R (B220; 1:50; BD Pharmingen), rb anti-Desmin (1:100; Abcam), rb anti-Albumin, rat anti-CD11b (1:100; Abcam), rat anti-CD68 (1:200; AbDSerotec, Oxfordshire, UK), rat anti-MHCI (1:100; Abcam), rb anti-Ki67 (1:100; ThermoFisher Scientiﬁc), gt anti-HIF1α (1:50; R&D Systems), rb anti-BNIP3 (1:100; Abcam), rb anti-PDL1 (1:100; Abcam), rb anti-IFNγ (1:100; Abcam), rb anti-FOXP3 (1:50; Abcam), gt anti-mouse CCR2 (1:100; Abcam), mouse anti-human CCR2 (1:50; Abcam). After 2 h incubation, sections were washed two times with PBS/0.5% Casein. Secondary antibodies were applied to sections: anti-rb Dylight488, anti-rb AF488, anti-gt Dylight488, anti-rat FITC, anti-rat Cy3, anti-rb Cy3, anti-gt Cy3, anti-mouse Rhodamine Red, anti-rb AF647 or anti-gt AF647 (1:200; Dianova, Hamburg, Germany). Tissue was subsequently incubated for 1.5 h. Sections were washed with PBS and water. Finally, slices were covered with DAPI-containing mounting medium (Dianova). For staining of Desmin, before blocking step, sections were ﬁxed 10 min at −20 ◦C with Methanol, and for detection of CCR2 on mouse sections Antigen Retrieval Reagent (R&D Systems, Minneapolis, MN, USA) was used for 30 s. For staining of HIF1α, BNIP3, PDL1, IFNγ, and FOXP3, sections were permeabilized by 0.1% Triton X-100 (Sigma-Aldrich) in PBS or TBS (FOXP3) for 10 min. The staining of FOXP3 was performed by using TBS instead of PBS in all steps of the procedure. For detection of apoptotic cells, sections were stained with Apoptosis Detection Kit (ApopTag®, Merck Millipore, Burlington, MA, USA), based on TUNEL-technique, according to manufacturer protocol. Finally, IBA1-staining was performed as described. For detection of apoptotic cells, sections were stained with Apoptosis Detection Kit (ApopTag®, Merck Millipore, Burlington, MA, USA), based on TUNEL-technique, according to manufacturer protocol. Finally, IBA1-staining was performed as described. Human brain sections were treated with Autoﬂuorescence Eliminator Reagent (Millipore) accordingly manufacturer’s instructions before blocking. Human brain sections were treated with Autoﬂuorescence Eliminator Reagent (Millipore) accordingly manufacturer’s instructions before blocking. Images were acquired by ﬂuorescence microscope Zeiss Axio Observer Z1 (Zeiss MicroImaging GmbH) or confocal microscope SP8 (Leica Biosystems, Nussloch, Germany). Images were acquired by ﬂuorescence microscope Zeiss Axio Observer Z1 (Zeiss MicroImaging GmbH) or confocal microscope SP8 (Leica Biosystems, Nussloch, Germany). Sections were analyzed systematically. 4.5. Lectin Application Mice were anesthetized, and 0.1 mL of 1 mg/mL lectin (Dylight488 Lycopersicon Esulentum (Tomato) Lectin; Vector Laboratories, Burlingame, CA, USA) was slowly injected via tail vein. After 5 min, mouse was perfused with 4% paraformaldehyde (PFA, Sigma-Aldrich, St. Louis, MO, USA). 14 of 19 Cancers 2020, 12, 1882 14 of 19 4.7. Immunohistochemistry For hematoxylin-eosin-staining, frozen sections were ﬁxed in absolute ethanol, dyed with hematoxylin and subsequently rinsed with water. Afterwards, sections were dyed in eosin, followed by a short wash in tap water. Then, ascending ethanol series and ﬁnally, xylene was used for tissue dehydration. The Zeiss Axio Observer Z1 Microscope (Zeiss MicroImaging GmbH, Jena, Germany) was used for taking mosaic pictures. 4.6. Cardial Brain Perfusion To prepare tissue for histological analysis, at day 21 animals were anesthetized. PBS or PFA were used for perfusion into the left heart-ventricle. Brains were directly used for the preparation of brain cell suspensions or post-ﬁxed in 4%-PFA and dehydrated in ascending sucrose solution (10%/20%/30%). Frozen brains were processed in a microtome. 10 µm or 50 µm sections were prepared and stored at −80 ◦C. 4.9. Cell Homogenization On day 21, mice were perfused with 10 mL PBS, brains extracted and cerebella, as well as olfactory bulbs, removed. Brains of mice or 400 mg human brain tissue were minced and digested in enzyme mixtures from the Papain Neuronal Tissue Dissociation Kit (Miltenyi Biotec, Bergisch Gladbach, 15 of 19 Cancers 2020, 12, 1882 Germany) according to the manufacturer´s protocol. Cell suspensions were ﬁltered (Pre-separation ﬁlter, 30 µm, Miltenyi Biotec). Homogenates were used either for direct ﬂow cytometric analyses or speciﬁc CD11b+ cell isolation. Germany) according to the manufacturer´s protocol. Cell suspensions were ﬁltered (Pre-separation ﬁlter, 30 µm, Miltenyi Biotec). Homogenates were used either for direct ﬂow cytometric analyses or speciﬁc CD11b+ cell isolation. 4.11. Isolation of Bone Marrow Cells and Generation of Macrophages 4.11. Isolation of Bone Marrow Cells and Generation of Macrophages Femurs of WT and Ccr2-/- mice were rinsed with PBS. Following washing, the cells were plated into a cell culture ﬂask with RPMI medium 1640 (ThermoFisher Scientiﬁc; including 10% FCS, 1% penicillin/streptomycin) and 20 ng/mL recombinant M-CSF (Miltenyi Biotec). The medium was changed every second day for eight days. Cells were detached by a cell scraper and used for culture experiments. Macrophages with a purity of 90% were obtained. 4.12. RNA Isolation Real-Time PCRs The isolated CD11b+ cells of murine and human brains were used for RNA extraction via PureLink™RNA Mini Kit (Invitrogen™, Carlsbad, CA, USA) according to the manufacturer’s instructions. Here, CD11b+ cells of six murine brain tumor hemispheres or three naïve brains were pooled to generate one RNA sample. The quality of RNA was veriﬁed with Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Reverse transcription was performed with puriﬁed RNA using QuantiTect Reverse Transcription Kit (Qiagen, Hilden, Germany) according to the manufacturer instructions. For semi-quantitative real-time PCR, reactions were performed in triplicates in a volume of 25 µL, containing 12.5 µL of SYBR Premix Ex Taq Kit (Takara Bio Inc., Kusatsu, Japan), forward and reverse primers, following the manufacturer’s protocol. Used primers are listed in Supplementary Table S1. Expression levels were normalized to 18S mRNA levels. Relative expression levels were determined by the ∆∆Ct method. 4.10. Isolation of CD11b+ Cells Following mechanic and enzymatic digestion of mouse and human brain tissues, cells were labeled with anti-CD11b magnetic microbeads (Miltenyi Biotec). The cell suspension was added to a MACS LS column (Miltenyi Biotec), which was placed in a magnetic ﬁeld. Non-labeled cells were washed out, and CD11b positive cells were eluted. This procedure was repeated with an MS column (Miltenyi Biotec). Positive cells with high purity (>95%) were achieved. 4.13. Culture of Myeloid Cells In Vitro and Immunocytochemistry For the cultivation of microglia following isolation by anti-CD11b microbeads from naïve mouse brains, cells were seeded at a density of 2 × 105 cells/well in 8 well glass bottom chamber slides (ThermoFisher Scientiﬁc) and cultured in tumor-conditioned DMEM medium for seven days. The medium was changed every second day. For the cultivation of macrophages, cells were seeded at a density of 1.2 × 105 cells/well in 8 well glass bottom chamber (ThermoFisher Scientiﬁc) and cultured in tumor-conditioned RPMI medium 1640 for four days. The medium was changed every second day. The tumor-conditioned medium was obtained by collecting the supernatant of GL261 cells which were grown for three days to 80% conﬂuence in DMEM or RPMI 1640. The supernatant was centrifuged two times by maximum speed to exclude glioma cells. Immunocytochemistry of microglia and macrophages was performed after ﬁxation of the cells with 4% paraformaldehyde in PBS for 20 min. Subsequently, microglia were permeabilized for 10 min with 0.3% Triton X-100 in PBS and blocked in 1% Casein/PBS for 1 h. Primary antibodies were incubated 2 h at room temperature: rat anti-CD11b (1:100; Abcam), rat anti-CD68 (1:100; AbDSerotec), and rabbit anti-IBA1 (1:200; WAKO). For visualization, cells were incubated with ﬂuorochrome-conjugated secondary antibodies (anti-rabbit Dylight488 and anti-rat Cy3; Dianova) for 1.5 h protected from light. The macrophages were treated for 10 min with methanol at −20 ◦C and blocked in 1% Casein/PBS for 30 min. Primary antibodies were incubated 2 h at room temperature: rat anti-F4/80 (1:100; Abcam) and Cancers 2020, 12, 1882 16 of 19 16 of 19 goat anti-IBA1 (1:100; Abcam). As secondary antibodies, anti-rat Cy3 and anti-goat AF647 (Dianova) were used. After staining, chambers were removed and slides were mounted with DAPI-containing mounting medium (Dianova). Images were acquired at 20× magniﬁcation using an inverse ﬂuorescence microscope Zeiss Axio Observer Z1 (Carl Zeiss MicroImaging GmbH). 4.14. Flow Cytometry To assess immune cells, cells were washed with PBS/0.5% BSA. Staining was performed with PE anti-CD11b (BD Pharmingen) and APC anti-CD45 (BD Pharmingen), or FITC anti-CD11b (1:100; BD Pharmingen), PE anti-CCR2 (1:20; R&D Systems) and APC anti-CD45 (1:200). Isotype control rat IgG2b (R&D Systems) was used for CCR2-antibody. Subsequently, cells were incubated for 20 min on ice. DAPI was added to detect dead cells. For other surface molecules, cells were ﬁxed. Here, the cell suspension was washed with PBS, ﬁxed for 20 min with 2% PFA, and washed with PBS/0.5% BSA. Cells were permeabilized with 0.5% saponin (exception MHCII, PBS/0.5% BSA) and staining was performed in 0.5% saponin (MHCII only 0.5% BSA/PBS). Antibodies PE anti-CD11b (1:200) and APC anti-CD45 (1:200) were used in combination with the respective speciﬁc antibodies coupled to FITC: anti-I-Ab (1:50), anti-CD80 (1:100), anti-CD86 (1:100) as well as isotype controls rat IgG2a, Armenian hamster IgG and mouse IgG2a (BioLegend, San Diego, CA, USA). Cells were incubated for 20 min at room temperature. Cells were washed with 0.5% saponin and analyzed by ﬂow cytometry. All samples were analyzed with BD FACS Canto II (BD Pharmingen) and evaluated with FlowJo software (Ashland, OR, USA). 4.15. Statistical Analysis All data presented with mean and error bars indicate standard deviation. GraphPad Prism Software (San Diego, CA, USA) was used to calculate statistical signiﬁcance. Diﬀerences were evaluated by two-tailed unpaired Student´s t-test or as indicated. Signiﬁcant values were presented as p < 0.05. References 1. Louis, D.N.; Ohgaki, H.; Wiestler, O.D.; Cavenee, W.K.; Burger, P.C.; Jouvet, A.; Scheithauer, B.W.; Kleihues, P. The 2007 WHO classiﬁcation of tumours of the central nervous system. Acta Neuropathol. 2007, 114, 97–109. [CrossRef] 2. Ohgaki, H.; Kleihues, P. Epidemiology and etiology of gliomas. Acta Neuropathol. 2005, 109, 93–108. [CrossRef] 3. Charles, N.A.; Holland, E.C.; Gilbertson, R.; Glass, R.; Kettenmann, H. The brain tumor microenvironment. Glia 2011, 59, 1169–1180. [CrossRef] 4. Roggendorf, W.; Strupp, S.; Paulus, W. Distribution and characterization of microglia/macrophages in human brain tumors. Acta Neuropathol. 1996, 92, 288–293. [CrossRef] 5. Watters, J.J.; Schartner, J.M.; Badie, B. Microglia function in brain tumors. J. Neurosci. Res. 2005, 81, 447–455. [CrossRef] 6. Kushchayev, S.V.; Kushchayeva, Y.S.; Wiener, P.C.; Scheck, A.C.; Badie, B.; Preul, M.C. Monocyte-derived cells of the brain and malignant gliomas: The double face of Janus. World Neurosurg. 2014, 82, 1171–1186. [CrossRef] 7. Müller, A.; Brandenburg, S.; Turkowski, K.; Müller, S.; Vajkoczy, P. Resident microglia, and not peripheral macrophages, are the main source of brain tumor mononuclear cells. Int. J. Cancer 2015, 137, 278–288. [CrossRef] 8. Badie, B.; Schartner, J.; Klaver, J.; Vorpahl, J. In vitro modulation of microglia motility by glioma mediated by hepatocyte growth factor/scatter factor. Neurosurgery 1999, 44, 1077–1082. [CrossRef] mediated by hepatocyte growth factor/scatter factor. Neurosurgery 1999, 44, 1077–1082. [CrossRef] 9. Zhai, H.; Heppner, F.L.; Tsirka, S.E. Microglia/macrophages promote glioma progression. Glia 2011, 59, 472–485. [CrossRef] Zhai, H.; Heppner, F.L.; Tsirka, S.E. Microglia/macrophages promote glioma progression. Glia 2011, 59 472–485. [CrossRef] 10. Ellert-Miklaszewska, A.; Dabrowski, M.; Lipko, M.; Sliwa, M.; Maleszewska, M.; Kaminska, B. Molecular deﬁnition of the pro-tumorigenic phenotype of glioma-activated microglia. Glia 2013, 61, 1178–1190. [CrossRef] 11. Komohara, Y.; Horlad, H.; Ohnishi, K.; Fujiwara, Y.; Bai, B.; Nakagawa, T.; Suzu, S.; Nakamura, H.; Kuratsu, J.; Takeya, M. Importance of direct macrophage-tumor cell interaction on progression of human glioma. Cancer Sci. 2012, 103, 2165–2172. [CrossRef] 12. Markovic, D.S.; Vinnakota, K.; Chirasani, S.; Synowitz, M.; Raguet, H.; Stock, K.; Sliwa, M.; Lehmann, S.; Kalin, R.; van Rooijen, N.; et al. Gliomas induce and exploit microglial MT1-MMP expression for tumor expansion. Proc. Natl. Acad. Sci. USA 2009, 106, 12530–12535. [CrossRef] p 3. Galarneau, H.; Villeneuve, J.; Gowing, G.; Julien, J.P.; Vallieres, L. Increased glioma growth in mice depl of macrophages. Cancer Res. 2007, 67, 8874–8881. [CrossRef] 14. 5. Conclusions In our study of a murine glioma model, the reduced accumulation of TAMs in Ccr2-deﬁcient mice is accompanied by increased tumor volumes based on the enhanced proliferation of glioma cells, changes in their inﬂammatory gene expression and improved integrity of tumor blood vessels. Notably, approximately 70% of TAMs inﬁltrated glioma tissues independently of the CCR2/CCL2 signal which underlines the role of other important recruitment factors. Supplementary Materials: The following are available online at http://www.mdpi.com/2072-6694/12/7/1882/s1, Figure S1: Inﬁltration of T cells into tumor tissues is unaﬀected by Ccr2-deﬁciency but B cell counts are reduced, Figure S2: Myeloid cells of naïve Ccr2-deﬁcient mice show same morphology and marker expression as the wildtypes, Figure S3: BNIP3 is localized in the nucleus and expressed by tumors of the wildtype and Ccr2KO mice, Figure S4: Vascular structure of brains is similar between mouse strains. Table S1: Primer sequences, Table S2: Basic expression of molecules. Author Contributions: Conceptualization, M.F., P.V. and S.B.; investigation, M.F., A.B., A.D.B., A.M., A.G., I.K., K.T., L.S., J.R. and S.B.; resources, O.R. and T.B.; data curation, M.F. and S.B.; writing—original draft preparation, M.F. and S.B.; writing—review and editing, A.B., A.D.B., A.M., T.B., K.T., L.S. and P.V.; visualization, M.F. and S.B.; supervision, P.V. and S.B.; project administration, S.B.; funding acquisition, P.V. All authors have read and agreed to the published version of the manuscript. Author Contributions: Conceptualization, M.F., P.V. and S.B.; investigation, M.F., A.B., A.D.B., A.M., A.G., I.K., K.T., L.S., J.R. and S.B.; resources, O.R. and T.B.; data curation, M.F. and S.B.; writing—original draft preparation, M.F. and S.B.; writing—review and editing, A.B., A.D.B., A.M., T.B., K.T., L.S. and P.V.; visualization, M.F. and S.B.; supervision, P.V. and S.B.; project administration, S.B.; funding acquisition, P.V. All authors have read and agreed to the published version of the manuscript. Funding: This research was funded by the German Research Foundation (DFG; SPP1190 and Excellence Cluster NeuroCure). A.M. was funded by a doctoral scholarship from the Sonnenfeld foundation. Acknowledgments: We acknowledge support from the German Research Foundation (DFG) and the Open Access Publication Fund of Charité—Universitätsmedizin Berlin. The authors thank Susanne Mueller (Charité Core Facility 7T Experimental MRIs, Charité University Medicine Berlin) for her assistance in MRI measurement. Additionally, we thank Sabine Seidlitz and Melina Nieminen-Kelhä for organizational support. We thank Mathias Heikenwälder (HMGU Munich and DKFZ Heidelberg, Germany) for providing the initial Ccr2-/- mice. 5. Conclusions 17 of 19 Cancers 2020, 12, 1882 Conﬂicts of Interest: The authors declare no conﬂict of interest. 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https://openalex.org/W1484878777_2	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	1,942	4,802	php?option=com_content&task=view&id=117&Itemid=29 http://imgranados.wordpress.com/ Modelo de comunicaciones digitales (Octavio Islas) para http://redalyc.uaemex.mx/pdf/649/64911103.pdf ambientes comunicativos glocales, en el imaginario de la sociedad de la ubicuidad Modelo conceptual (de una sociedad en red) http://www.utwente.nl/cw/theorieenoverzicht/Theory%20clusters/Communication%20Processes/Network%20Theory%20 and%20analysis_also_within_organizations-1.doc/ Web 2.0 Framework http://rossdawsonblog.com/weblog/archives/2007/05/launching_the_w.html Web 2.0 Landscape http://rossdawsonblog.com/weblog/archives/2007/05/launching_the_w.html El futuro del ciclo de vida de los media http://rossdawsonblog.com/weblog/archives/2008/07/ launch_of_the_f.html Futuro de los medios: herramientas estratégicas http://rossdawsonblog.com/weblog/archives/2008/07/ launch_of_futur.html Tendencias en la vida en redes http://rossdawsonblog.com/weblog/archives/2006/12/trend_ map_for_2.html Cuadrantes de comunicación y colaboración conversacional http://www.brooksandrus.com/blog/2008/05/05/influencersand-social-media/ Social capital of Blogspace - Ross Mayfield http://picsicio.eu/domain/relationship-economy.com/ Mapa de redes sociales de David Pollar http://blogs.salon.com/0002007/2005/11/02.html#a1327 El prisma de conversación digital de Brian Solis y Jesse Thomas http://adolescentesantelosmedios.blogspot.com/2009/07/ la-comunicacion-adolescente-se.html Captology http://edweb.sdsu.edu/people/ARossett/pie/Interventions/ captology_2.htm Nuevo modelo de comunicación http://tumurodigital.wordpress.com/ Impacto de las nuevas tecnologías 2.0 http://cursos.cepcastilleja.org/mod/forum/discuss. php?d=5670 El poder de las herramientas 2.0 en empresas e instituciones http://www.fernandarubio.com.ar/archives/1433 Mapa de medios sociales en España Modelo estratégico SMM http://protocoloycomunicacion.blogspot.com/2009_02_01_ archive.html http://isaacvidal.blogspot.com/ Modelo de Universidad + comunicación corporativa tradi- http://simbiodiversidad.blogspot.com/2009/10/medioscional sociales-y-cultura-digital.html Universidad + comunicación digital http://simbiodiversidad.blogspot.com/2009/10/mediossociales-y-cultura-digital.html Modelo de estrategias de comunicación digital en las insti- http://nomada.blogs.com/jfreire/publicaciones/ tuciones culturales Difusión como parte de la producción http://nomada.blogs.com/jfreire/2009/07/herejas-de-losnuevos-procesos-culturales.html Campaña de medios sociales de Gary Hayes and Laurel http://www.flickr.com/photos/garyhayes/2973684461/ Papworth 2008 Modelo de Ross Dawson encuadre de estrategia de medios http://www.rossdawsonblog.com/weblog/archives/2009/07/ launch_of_socia.html Anagramas, Volumen 9, Nº 18, pp. 93-110 - ISSN 1692-2522 - enero-junio de 2011. 210 p. Medellín, Colombia 105 Andrés Felipe Giraldo Dávila Nuevo modelos de comunicación Dirección URL Modelo de Chaffey y Smith sobre categorías de comunicación http://emarketingacademia.com/ electrónicas Involve Introduction http://www.dload.com.au/Involve/Introduction Mapa interactivo de conocimiento Modelo integrado de comunicación 360 ° http://semanticstudios.com/publications/semantics/000006. php http://blog.mattiaskindell.com/page/4/ Expansión de Brief de RRPP http://prwarrior.typepad.com/my_weblog/about-me.html. Negocios y redes sociales http://www.soravjain.com/2009/12/social-business-modelcollated-ideas.html http://www.nathaliemagniez.com/ecosystems/businessecosystem-instead-of-business-model/ http://www.headland.co.uk/pages/brand-development Ecosistema de negocios Diagrama estratégico digital Cómo ayudan las redes sociales http://www.compromisoempresarial.com/rsc/multimedia/2010/06/como-ayudan-las-redes-sociales/ PKI, Public Key Infrastructure (Infraestructura de clave pública) http://unesrinfe5.blogspot.com/ Modelo Posting en los social media marketing http://www.soravjain.com/2009/11/posting-month-how-tocreate-right-kind.html http://seoamit.wordpress.com/2009/02/10/roi-model-forsocial-media/ http://reddigital.cnice.mecd.es/2/james/01James_3.html Modelo de los medios sociales ROI Modelo del alumno digital y el nuevo profesor Modelo de esquema guía de Internet para comercio elec- http://www.programadorphp.org/blog/recursos-softwaretrónico avanzados-para-empresas/ El modelo organizativo en red http://pedropabloramos.nireblog.com/ Gráficos de comunicación corporativa http://comunisfera.blogspot.com/2008/01/blog-post.html http://3.bp.blogspot.com/_HF-3vmiEjpo/R57xGMLmSDI/ AAAAAAAAAGE/0P1PrALwlzI/s1600-h/nuevo-esquema.jpg http://expressivitytheory.com/communication_model.html Modelo de comunicación digital Elementos básicos del sistema de comunicación digital Knowledge Visualization http://wirelesscafe.wordpress.com/2009/07/10/tutorial-ibasic-elements-of-digital-communication-system/ http://www.idiagram.com/ideas/knowledge_visualization.html The geosocial universo h t t p : / / w w w. i n s i d e f a c e b o o k . c o m / w p - c o n t e n t / uploads/2010/08/082010-rdup-social-sp.jpg Hypodermic to networked models http://cb3blog.wordpress.com/category/political-communications/ Integration of Online and Offline Media/Communication http://megoyanagi.thoseinmedia.com/2010/04/03/volChannels 26-%E2%80%93-how-to-effectively-integrate-mediacommunication-channels-part-2/ Keys for Social Media success: http://michaelcnewhouseblog.blogspot.com/2010_04_01_archive.html Receiving messages http://www.daisyrust.com/category/digital-art-hacks/ Resale of advertising spaces Marketing mix Unified communications http://digital-signage-software.net/2009/11/02/roi-digitalsignage-networks/ http://katecarruthers.com/blog/2009/02/marketing-principlesfor-the-digital-age/ http://www.processflows.co.uk/vertical-markets/public-sector/ Fuente: elaboración propia. 106 ANAGRAMAS ‘Realidad virtual’: análisis del marco teórico para explorar nuevos modelos de comunicación Conclusiones Los 53 modelos de comunicación encontrados en la primera pesquisa del proyecto de investigación “Nuevos modelos de comunicación” permiten describirse desde una o varias de las tres posturas replanteadas en el marco teórico. Algunos modelos miden el vínculo entre la plataforma virtual con la realidad; otros analizan las relaciones dadas entre los procesos informáticos y las ciencias humanas; y, en alguna proporción, los esquemas reflexionan sobre la transformación del dilema público-privado en una sociedad conectada por la virtualidad. El replanteamiento teórico del proyecto investigativo reemplazó la prospectiva etnográfica que desde un comienzo se había trazado como metodología. Enfatizar que no es necesario hacer encuestas para diseñar modelos de comunicación sino revisar información derivada de la misma virtualidad, entregó una mejor orientación al objetivo inicial de la investigación que giró en torno a medir si la interacción virtual de los individuos afecta la comunicación cara a cara. Retortillo (2000) argumenta desde Ortega y Gasset que en la actualidad la tecnología es el ‘medio natural’ de la humanidad donde las modificaciones y avances de la una transforman y modifican la otra. Luego, los cambios tecnológicos hacen que la naturaleza del ser humano también sufra modificaciones El propósito de los nuevos modelos de comunicación es analizar la importancia que los procesos de transformación tecnológica tienen sobre las interacciones humanas, tanto en el ámbito privado como en los asuntos públicos, donde lo virtual ha trastocado sustancialmente los conceptos de espacialidad, geografía y temporalidad. Modelar la comunicación no pende de una encuesta o sondeo de opinión, sino de las emergentes tendencias que la misma virtualidad define a través de la revisión de información y el análisis descriptivo de fuentes emanadas de Internet, tal y como benéficamente se obtuvo en varios meses de análisis de información dentro de la misma plataforma virtual. Para la revisión de nuevos modelos de comunicación se tuvieron en cuenta elementos como la realidad virtual y la sociabilidad privada para medir aspectos comunicativos, públicos o privados, derivados de la utilización de herramientas virtuales, los cuales permiten a investigadores y profesionales de la comunicación teorizar, analizar y planificar. La virtualidad y la realidad no son dos mundos diferentes sino parámetros diferenciadores que miden la relación entre individuos en la vida social. Ni lo virtual representa todas las estructuras humanas ni es la única dimensión válida como realidad en un mundo inundado por las tecnologías de la información. Las condiciones de tiempo y espacio deben ser revalidadas con la enorme influencia de la tecnología en asuntos como la vida privada, el trabajo, la productividad y la educación. El mundo virtual y la realidad convergen en una misma esfera donde se interpretan símbolos y significados para la interacción humana. La tecnología incrementa el rendimiento del hombre y propicia la innovación, la competitividad y las competencias, además de determinar los comportamientos en diversos contextos sociopolíticos. Las relaciones humanas están intervenidas por las actualizaciones progresivas de la tecnología que conduce un nuevo pensamiento para conducir diversas conjeturas conceptuales en el campo de estudio de la comunicación y sus vertientes como la publicidad, el mercadeo, el periodismo, las organizaciones y las relaciones públicas. Barbieri (2004) analiza que los nuevos modelos de comunicación emergen a partir del uso de herramientas virtuales y su enfoque derivado de la ‘Teoría de la Interface’, la cual se refiere a que la interface más importante en el actual contexto de la comunicación es el encuentro entre el hombre y la máquina. De allí parte un primer embrión de la comunicación, donde la virtualidad deriva otras formas de mediación a través de diversos procesos de información y confección de mensajes que a la postre comunican. El uso de hardware y software –puntualiza Barbieri- crea una interface cuando pasa del procesamiento de información a las interacciones. Dicha perspectiva teórica permitirá entrelazar aspectos de la ingeniería junto a los de la comunicación, ya que dispositivos, programación y aplicativos tienen inferencia en Anagramas, Volumen 9, Nº 18, pp. 93-110 - ISSN 1692-2522 - enero-junio de 2011. 210 p. Medellín, Colombia 107 Andrés Felipe Giraldo Dávila las formas en que la sociedad y sus individuos se comunican. Una contracción paradigmática está cuando el hombre, absorbido por la máquina, limita sus capacidades de comunicarse cara a cara con sus semejantes, asunto que desborda los objetivos trazados en la investigación pero que sería materia de reflexión en diversos campos de las ciencias humanas. Las TIC crean nuevos espacios de interacción donde las relaciones desestiman el valor de la espacialidad física reemplazada por un escenario interactivo que modifica la percepción real de lo físico. Para ejercer algún trabajo, realizar alguna transacción bancaria, promover un líder político o entablar una cita amorosa ya no se requiere de factores geográficos sino virtuales, que afectan a posteriori las relaciones que se dan en tiempo y espacio real. La telefonía móvil, las cámaras fotográficas, las descargas de música, las herramientas de office, los programas de diseño son dispositivos que generan productos para ser aplicados en un escenario virtual carburado por un ordenador y permiten conectar al individuos en escenarios virtuales privados o públicos que se pueden analizar por la teoría de la interface. La interface virtual conecta a las personas con las máquinas en un dispositivo que posibilite la construcción de comunidades, grupos u organizaciones, las cuales se ven altamente influidas en sus interacciones por el aporte que la virtualidad ofrece. La realidad de la humanidad, es decir, la virtualidad, posibilita el análisis descriptivo de nuevos modelos de comunicación que ofrezcan una representación del mundo. Usar el ciberespacio para asumir la realidad del mundo apunta a fortalecer las interacciones humanas y una mejor comprensión del mundo. El hombre debe desarrollar capacidades, habilidades y hábitos en el uso de herramientas virtuales para jugar un papel activo en una sociedad de la información donde el conocimiento es un valor agregado y de competitividad. Quizá hoy no es necesario agradecer a quienes hicieron de este mundo virtual-real una posibilidad, ya que la sociedad ha asumido estos cambios como un proceso natural en sí mismo. 108 Alexandre Hohagen, Presidente de Google para América Latina enfatiza en lo siguiente: Imagínese lo que era su vida hace poco más de 10 años, sin Google, iPod, YouTube, Facebook, Amazon, los smartphones y la televisión digital. A pesar de los grandes cambios que estas nuevas tecnologías aportan a nuestra vida, la verdad es que todavía es incierta la transformación y el legado que nos dejarán (Portafolio, enero 21 de 2011, p.31) Roy Amara, investigador y científico de la Universidad de Stanford, citado por Hogagen sentencia: “Tendemos a sobre estimar el impacto de las nuevas tecnologías en el corto plazo y a subestimar sus efectos en el largo plazo” (Ibíd.) Las prioridades de la agenda pública, los estudios investigativos de la comunicación, la decantación de la sociedad del conocimiento y la innovación apuntan hacia el impacto de las transformaciones tecnológicas y sus usos prácticos, por lo que ese ámbito de la virtualidad necesariamente debe apuntar a una utilidad en lo tangible, es decir, la realidad. Referencias bibliográficas Alsina, M. (2001): Teorías de la comunicación: ámbitos, métodos y perspectivas. Universidad Autónoma de Barcelona, Universidad Jaime I, Universidad Pompeu Fabra, Universidad de Valencia. Amor, D. (2002): The e-bussiness (r) evolution. New Jersey: Prentice Hall. Barbieri, D (2004): “Fearful Simmetry: reflexiones sobre el concepto de interface”. Razón y Palabra, No. 38, abril-mayo [en línea] http://www.razonypalabra.org. mx/anteriores/n38/dbarbieri.html Recuperado el 01/07/2010 19:23 hs. Botero Montoya, L. (2007): Teoría de públicos: lo público y lo privado en la perspectiva de la comunicación. Medellín: Sello Editorial Universidad de Medellín. Cabero, J. (2001): Tecnología educativa: diseño y utilización de medios en la enseñanza. Barcelona: Paidós. Echeverría, J. (1999): Los señores de aire: telépolis y el tercer entorno. Barcelona: Destino. ANAGRAMAS ‘Realidad virtual’: análisis del marco teórico para explorar nuevos modelos de comunicación García Blanco, J. (2002): Virtualidad, realidad, comunidad. Un comentario sociológico sobre la semántica de las nuevas tecnologías, Papers No.68 pp. 81-106. Hohagen, Alexandre (2011): La década que no se termina. Portafolio. Bogotá: enero 21 de 2011, p.31. Islas, O. (2003): El desarrollo tecnológico como detonador de algunas crisis epistemológicas que enfrentan las ciencias de la comunicación. En: Razón y Palabra, No. 33, junio-julio [en línea] http:/ www.razonypalabra.org.mx/anteriores/n34/oislas. html Consultado el 29/06/2010 08:32 hs. López García, G (2005): Modelos de comunicación en Internet. Valencia: Tirant Lo Blanch. Martínez, G. (2001): Lo público y lo virtual en escena, en: Revista Venezolana de Gerencia, octubre/diciem- bre, año/vol. 6, No.16, pp. 573-592. Universidad del Zulia. Retortillo Franco, F. (2000): Realidad y virtualidad: ¿Verdadera dicotomía en el entorno de las tecnologías de la información y la comunicación? En: AA. VV., Nuevas tecnologías, viejas esperanzas: las nuevas tecnologías en el ámbito de la discapacidad y las necesidades educativas especiales, pp: 257-262, I Congreso Internacional de Nuevas Tecnologías y Necesidades Educativas Especiales, Murcia. Siles González, I. (2005) Internet, virtualidad y comunidad. Revista Ciencias Sociales, No. 108:55-69. Wolton, D. (2000) Internet ¿Y después? Barcelona: Gedisa. Anagramas, Volumen 9, Nº 18, pp. 93-110 - ISSN 1692-2522 - enero-junio de 2011. 210 p. Medellín, Colombia 109.	11,359
W2944003723.txt_1	Open-Science-Pile	Open Science	Various open science	2,018	Discovering the deep: Exploring remote Pacific marine protected areas	None	English	Spoken	37	85	Oceanography Vol. 31, No. 1, Supplement, March 2018 New Frontiers in Ocean Exploration The E/V Nautilus, NOAA Ship Okeanos Explorer, and R/V Falkor 2017 Field Season GUEST EDITORS \| Nicole A. Raineault, Joanne Flanders, and Amya Bowman.	27,882
W2086029987.txt_1	German-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	829	1,721	Document generated on 07/14/2024 12:46 a.m. Laval théologique et philosophique BRUGGER, Walter, Der dialektische Materialismus und die Frage nach Gott Jaromír Danek Volume 37, Number 3, 1981 Hegel (1831-1981) URI: https://id.erudit.org/iderudit/705890ar DOI: https://doi.org/10.7202/705890ar See table of contents Publisher(s) Faculté de philosophie, Université Laval ISSN 0023-9054 (print) 1703-8804 (digital) Explore this journal Cite this review Danek, J. (1981). Review of [BRUGGER, Walter, Der dialektische Materialismus und die Frage nach Gott]. Laval théologique et philosophique, 37(3), 377–377. https://doi.org/10.7202/705890ar Tous droits réservés © Laval théologique et philosophique, Université Laval, 1981 This document is protected by copyright law. Use of the services of Érudit (including reproduction) is subject to its terms and conditions, which can be viewed online. https://apropos.erudit.org/en/users/policy-on-use/ This article is disseminated and preserved by Érudit. Érudit is a non-profit inter-university consortium of the Université de Montréal, Université Laval, and the Université du Québec à Montréal. Its mission is to promote and disseminate research. https://www.erudit.org/en/ CO\Il'TES 1{f''if)IS simplicité et une sérénité remarquables, un message difficile à monnayer. L'A. ne prétend pas avoir résolu tous les problèmes que soulève la critique néorestamentaire; mais il donne confiance en la démarche que suit cette critIque et dans la qualité des résultats qu'elle atteint. "Ce livre est une invitation à la confiance, à la recherche ", disait l'A. tout au début de son ouvrage (p. 9); son invitation sera sans doute entendue d'un large public. Nous souhaitons que beaucoup de lecteurs puissent reprendre pour leur compte personnel la rétlcxion que l'A. livre au terme de son étude: ,,!\13 confiance dans les documents primaires de I~ foi chrétienne s'est trouvée fortifiée plutôt que secouée. L'érudition ne me donne pas la foi. mais clk augmente ma conviction que ma foi n'est pas mal placée" (p. 156). Paul-Émile LAN(;~VIN, S.J. déformation (anti-hégélienne, dis\lll'-l1ousj de 1" téléologje mondiale Dl! universelk, tran'jpo~ant J'interrogation ,ur les sources et les finalités de la nature et de l'histoire dans le domaine d'une ontologie naïvement positive. À travers les considératIons du livre. qUI tstle résultat d'une connaissance profonde de la littérature marxiste (contemporaine en partIculIer. voir l'index bibliographique), le lecteur attentif voudrait retrouver davantage la problématique de J'histoire en tant que révolutIon dramatique de la liberté, prohlématique opposée à l'a,,omalisme idéologique de l'athéisme mar:xistc-lélllniSlt' L'aspect éthique de la critiqUé du matériaiisille dialectique (inséparable de sa version dite" hi,torique »)) exigerait une confront:Jtion explicite avec l'humanisme chrétien et idéaliste en général Jaromir DA';lK Walter BR(;(;(;ER: Der dialektische Materialismus und die Frage nach Got!. (Le matérialisme dialectique ct la question de Dieu.) - J. Berchmam Verlag, Miinchen 1980. 254 p. (Collection: Wissenschaft und Gegenwart science et présent), La structure de cet ouvrage - le fond de l'étude très rigoureuse est nécessairement marqué par la normativité de la conception théologique du monde - est la suivante: Déterminations prélimlllaires - critiques des preuves de Dieu (kantienne et marxiste) - "matérialisme" - dialectique ·matérialiste. Les déterminations préliminaires svstématisent les concepts principaux tels que vérité, logique formelle et dialectique, praxis; principes ontologiques de la connaissance; matérialisme, réalisme, idéalisme, athéisme; conception scientifique du monde et idéologie, ... La position intuitive de ces concepts au début constitue cependant une faiblesse de la critique: ces concepts devraient achever l'analyse plutôt qu'être délimités par un apriorisme méthodique encore accentué par le fait qu'à la problématique du refus marxiste des preuves de Dieu est préalable la description de la critique kantienne du même domaine de la pensée. Suit le véritable noyau de l'étude: la question sur la matière pose le problème du monisme matérialiste dont l'absoluité évoque un pathos lypique pour un dogmatisme réduisant la richesse du monde objectivo-subjectif au mouvement des formes de la matIère. L'analyse de la dialectique matérialiste est à résumer comme démontrant la 377 Ei' COLLABORATtON, La teologia italiana oggL Facolta teologica dell' Italia settentriona!c, Milano, 1979.414 pages, 17 x 24 cm. Le présent volume est un hommage, à l'oeca",," de son 700 anniversaire de nalssance, à Mgr ( arlo Colombo, président de la Faculté de thé,)logie de l'Italie septentrionale, Il est fréquent de recueillir, en pareille occasion, en l'honneur de quelqu'un qui s'est distingué dans le domaine scientifique, la contribution de certains admirateurs. Les responsables de la présente publication ont préféré une autre solution, plus difficile, mais comhien plus valable, celle d'orienter en un dessein organique ces dIvers travaux. Il s'agissait, en somme, de rien moins que de faire l'histoire de la théologie contemporaine dont Mgr C. Colombo est indiscutablement un protagoniste. C'est avec des œuvres philosophiques, alors qu'il était professeur en cette discipline, quc Mgr C. Colombo commença sa carrière scientifïque au Séminaire de Venegono (1933-1938). Professeur de théologie dogmatique depuis 1938, il s'intéressa au problème de la guerre, de la reconstruction religieuse, sociale et politique de l'Italie. Son œuvre proprement théologique se situe entre les années 1946 et 1964 alors qu'il fut ordonné évêque titulaire de Vittoriana, le 7 mars 1964. Dès lors sa recherche théologique fit place aux aspects plutôt pastoraux de la théologie. L'étude en cours embrasse donc da,antage que les années COli vertes par l'activité théologique.	9,097
https://openalex.org/W2758803283	OpenAlex	Open Science	CC-By	2,017	Bit- and Power-Loading—A Comparative Study on Maximizing the Capacity of RSOA Based Colorless DMT Transmitters	Simon A. Gebrewold	English	Spoken	19,142	33,178	ETH Library Article Bit- and Power-Loading—A Comparative Study on Maximizing the Capacity of RSOA Based Colorless DMT Transmitters Simon Arega Gebrewold 1,2,, Romain Bonjour 1, Romain Brenot 3, David Hillerkuss 1,4 and Juerg Leuthold 1 Simon Arega Gebrewold 1,2,, Romain Bonjour 1, Romain Brenot 3, David Hillerkuss 1,4 and Juerg Leuthold 1 1 Institute of Electromagnetic Fields (IEF), ETH Zurich, 8092 Zurich, Switzerland; rbonjour@ethz.ch (R.B.); david.hillerkuss@huawei.com (D.H.); Juergleuthold@ethz.ch (J.L.) 1 Institute of Electromagnetic Fields (IEF), ETH Zurich, 8092 Zurich, Switzerland; rbonjour@ethz.ch (R.B.); david.hillerkuss@huawei.com (D.H.); Juergleuthold@ethz.ch (J.L.) 2 Now with RANOVUS GmbH, Nordostpark 07, 90411 Nuremberg, Germany 3 Thales Research and Technology and CEA Leti, Route de Nozay, a Joint Lab of Alcatel-Lucent Bell Labs France, III-V Lab, 91460 Marcoussis, France; romain.brenot@3-5lab.fr 4 Now with Huawei Technologies Duesseldorf GmbH, Optical & Quantum Laboratory, Riesstrasse 25-C3, 80992 Munich, Germany * Correspondence: simon@ranovus.com; Tel.: +49-176-9093-9230 R d 6 A 2017 A d 4 S b 2017 P bl h d 27 S b 2017 4 Now with Huawei Technologies Duesseldorf GmbH, Optical & Quantum Laboratory, Riesstrasse 25-C3, 80992 Munich, Germany y * Correspondence: simon@ranovus.com; Tel.: +49-176-9093-9230 Received: 6 August 2017; Accepted: 4 September 2017; Published: 27 September 2017 Abstract: We present a comparative study of the capacity increase brought by bit- and power-loading discrete multi-tone (DMT) modulation for low-cost colorless transmitters. Three interesting reﬂective semiconductor optical ampliﬁer (RSOA) based colorless transmitter conﬁgurations are compared: First, an ampliﬁed spontaneous emission (ASE) spectrum-sliced source; second, a self-seeded RSOA ﬁber cavity laser (FCL) and third, an externally seeded RSOA. With bit- and power-loaded DMT, we report record high line rates of 6.25, 20.1 and 30.7 Gbit/s and line rates of 4.17, 10.1 and 24.5 Gbit/s in a back-to-back and in a 25 km nonzero dispersion shifted ﬁber (NZDSF) transmission experiments for the three transmitter conﬁgurations, respectively. In all the experiments, BER (bit error ratios) below an FEC (forward error correction) limit of 7.5 × 10−3 were achieved. Keywords: bit- and power-loading; colorless transmitter; discrete multi-tone (DMT); reﬂective semiconductor optical ampliﬁer (RSOA); WDM-PON Author(s): This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. applied sciences 1. Introduction The scheme is low-cost and simple, but so far only bit rates below 1 Gbit/s have been achieved for each wavelength, with this scheme. The limitations of this scheme are high ﬁltering losses and susceptibility to chromatic dispersion due to a broad optical spectrum. p y p p p • Another cost-efﬁcient source—however with improved performance—is the self-seeded RSOA ﬁber cavity laser (RSOA-FCLs) as suggested in [20,21] and depicted in Figure 1b. In this case, a mirror is placed at the RN behind the WDM ﬁlter. This establishes a feedback mechanism to the RSOA to form a Fabry-Perot laser resonator. The operating wavelength of each ONU’s RSOA-FCL is determined by the WDM ﬁlter port it is connected to. This passive assignment of the emission wavelength simpliﬁes the wavelength control. Thus far, RSOA-FCL have shown to transmit up to 10 Gbit/s [22,23]. • The highest performance—however, a relatively expensive scheme—is the externally seeded RSOA, shown in Figure 1c. Here, a bank of lasers, one for each ONU, is installed at the CO. The WDM ﬁlter at the RN redistributes the individual cw (continuous wave) laser lines to each ONU. At the ONUs the RSOAs amplify the seeding light and encode the US signal. Externally seeded RSOAs have already shown transmission beyond 20 Gbit/s with pulse amplitude modulation (PAM) [24] and more than 25 Gbit/s using discrete multi-tone (DMT) [14,25]. However, in both cases, controlled offset ﬁltering by a WDM ﬁlter or an additional delay line interferometer was required, which makes the system sensitive to any wavelength drift. In addition, the extra lasers at the CO increases cost and energy consumption. In these three schemes, the modulation bandwidth limitation of RSOAs is a challenge that has to be addressed to fulﬁll the high data rate requirements of WDM-PONs. There have been different approaches to alleviate this issue. One solution is to improve the speed of the RSOAs by optimizing the physical design of the device [26]. Initial implementations of RSOA, mainly in the externally seeded scheme, employed non-return-to-zero (NRZ) amplitude-shift keying (ASK) modulation with either electronic [27,28] or optical signal processing [28,29] or a combination of electronic and photonic signal processing [30] to demonstrate 10 Gbit/s. Return-to-zero (RZ) and NRZ are compared in [31]. Advanced modulation formats have also been demonstrated for higher spectral efﬁciency with electronic and optical equalization in [12]. 1. Introduction Maximizing the bit-rate of colorless, inexpensive and “poorly” performing transmitters has become a research topic on its own. This is of particular importance in wavelength division multiplexed (WDM) passive optical networks (PONs) that are considered a possible solution to cope with the capacity surge in access networks [1,2]. WDM-PONs are being considered for applications requiring bidirectional high capacity and secure links. WDM-PONs are therefore ideal solutions to serve ﬁber to the home and building (FTTH/B) users with high capacity demands and also ideal to cover the needs of mobile fronthaul (ﬁber to the antenna, FTTA) [3,4]. The latter is characterized by small cell sizes with dense base stations. If WDM-PON are to be mass deployed, large quantity of wavelength speciﬁc optical network units (ONUs) would be required, which highly complicates the inventory. Thus, the challenge is to develop ONU transmitters that allow the installation of identical devices across all subscribers. The solution are colorless transmitters that operate across a broad wavelength range, while their speciﬁc operation wavelength is controlled by an external factor [5–8]. Installing identical colorless transmitters across all ONUs consequently, reduces the number of specialized components—thus simplifying the inventory. In addition, relatively low-quality mass producible colorless sources will make WDM-PON more economical. www.mdpi.com/journal/applsci Appl. Sci. 2017, 7, 999; doi:10.3390/app7100999 www.mdpi.com/journal/applsci Appl. Sci. 2017, 7, 999 2 of 17 For low-cost colorless transmitters, the speciﬁc operating wavelength can be controlled by two main methods. A ﬁlter at a remote location can automatically choose the transmission wavelength of an ONU [9,10]. Alternatively, colorless operation can also be implemented by using an external seed laser placed at the central ofﬁce (CO) to determine the upstream wavelength [11–16]. One promising colorless source is based on reﬂective semiconductor optical ampliﬁers (RSOAs). RSOAs are ideal sources as they offer a broad optical bandwidth at a low cost. There are three main architectures to integrate RSOAs in a WDM-PON as upstream (US) transmitters at the ONU. A theoretical and experimental comparison of the three architectures has been presented in [17]. • The simplest and most economical scheme is using RSOAs as ampliﬁed spontaneous emitters (ASE) source in a spectrum-sliced scheme, as shown in Figure 1a. The US signal is directly modulated onto the ASE of the RSOA. At the remote node (RN), a WDM ﬁlter slices the modulated ASE, passing only the assigned sub bands from each ONU to the central ofﬁce (CO) [18,19]. 1. Introduction In general, DMT has been used to maximize capacities of bandwidth limited intensity modulated direct detected (IM-DD) systems [25,34–36]. DMT is a type of OFDM with real-valued output signals that can be easily detected with inexpensive direct detection receivers. As a multicarrier format, DMT also brings the ﬂexibility of adapting the modulation format and power of any individual subcarrier to cope with the frequency dependence of the channel signal-to-noise ratio (SNR). In general, DMT has been used to maximize capacities of bandwidth limited intensity modulated direct detected (IM-DD) systems [25,34–36]. DMT is a type of OFDM with real-valued output signals that can be easily detected with inexpensive direct detection receivers. As a multicarrier format, DMT also brings the flexibility of adapting the modulation format and power of any individual subcarrier to cope with the frequency dependence of the channel signal-to-noise ratio (SNR). In general, DMT has been used to maximize capacities of bandwidth limited intensity modulated direct detected (IM-DD) systems [25,34–36]. DMT is a type of OFDM with real-valued output signals that can be easily detected with inexpensive direct detection receivers. As a multicarrier format, DMT also brings the ﬂexibility of adapting the modulation format and power of any individual subcarrier to cope with the frequency dependence of the channel signal-to-noise ratio (SNR). In this paper, we employ bit- and power-loaded DMT to show that the ASE spectrum-sliced scheme, the self-seeded RSOA-FCL, and the externally seeded RSOA configurations are viable US transmitters. The results are an expansion of our recent work [37], where only the maximized bit rates are presented. In this work, the SNR offered by each sources has been measured and compared. It is found that the SNR improves by more than 5 dB when switching from an ASE spectrum-sliced scheme to a self-seeded RSOA-FCL, and it improves by another ~5 dB when switching from a self- seeded RSOA-FCL to an externally seeded RSOA. Finally, transmission experiments have been performed. We demonstrate 6.25, 20.1, and 30.7 Gbit/s in back-to-back (BtB) and 4.17, 10.1 and 24.5 Gbit/s over 25 km NZDSF (non-zero dispersion shifted fiber) for the ASE spectrum-sliced, RSOA- FCL and externally seeded RSOA sources, respectively. No offset filtering was needed in any of these experiments [14 25] In this paper, we employ bit- and power-loaded DMT to show that the ASE spectrum-sliced scheme, the self-seeded RSOA-FCL, and the externally seeded RSOA conﬁgurations are viable US transmitters. 1. Introduction Wavelength division multiplexed passive optical network (WDM-PON) topologies with reflective semiconductor optical amplifier (RSOA) based colorless upstream (US) transmitters: (a) Amplified spontaneous emitter (ASE) spectrum-sliced transmitter: the US signal is directly modulated on to the ASE of the RSOA. The WDM filter at the remote node (RN) passes only the assigned sub band from each RSOA at the optical network units (ONUs); (b) Self-seeded RSOA-FCL (fiber cavity laser): a Fabry-Perot cavity indicated by the red dashed lines is formed between the RSOA and a mirror at the RN. The cavity is embedded in the distribution fiber (DF) of the PON; (c) Externally seeded RSOA: a laser at the central office (CO) seeds the RSOA at the ONU. The RSOA encodes the US signal onto the seed laser line. In all case, the ONUs are connected by the DF to the RN. The RN consists of the WDM filter and it is connected to the CO through the feeder fiber (FF) Figure 1. Wavelength division multiplexed passive optical network (WDM-PON) topologies with reﬂective semiconductor optical ampliﬁer (RSOA) based colorless upstream (US) transmitters: (a) Ampliﬁed spontaneous emitter (ASE) spectrum-sliced transmitter: the US signal is directly modulated on to the ASE of the RSOA. The WDM ﬁlter at the remote node (RN) passes only the assigned sub band from each RSOA at the optical network units (ONUs); (b) Self-seeded RSOA-FCL (ﬁber cavity laser): a Fabry-Perot cavity indicated by the red dashed lines is formed between the RSOA and a mirror at the RN. The cavity is embedded in the distribution ﬁber (DF) of the PON; (c) Externally seeded RSOA: a laser at the central ofﬁce (CO) seeds the RSOA at the ONU. The RSOA encodes the US signal onto the seed laser line. In all case, the ONUs are connected by the DF to the RN. The RN consists of the WDM ﬁlter and it is connected to the CO through the feeder ﬁber (FF). In general, DMT has been used to maximize capacities of bandwidth limited intensity modulated direct detected (IM-DD) systems [25,34–36]. DMT is a type of OFDM with real-valued output signals that can be easily detected with inexpensive direct detection receivers. As a multicarrier format, DMT also brings the flexibility of adapting the modulation format and power of any individual subcarrier to cope with the frequency dependence of the channel signal-to-noise ratio (SNR). 1. Introduction Polar RZ PAM4 with delay interferometer optical signal processing has achieved 20 Gbit/s [32]. Recently, multicarrier schemes such as adaptive orthogonal frequency division multiplexing (Adaptive-OFDM) [33] have been implemented for improved performance of the externally seeded scheme. Adaptive OFDM [25] and bit- and power-loaded DMT [14], both with optical equalization have already demonstrated beyond 25 Gbit/s transmission. 3 of 17 3 of 17 Appl. Sci. 2017, 7, 999 Appl. Sci. 2017, 7, 999 Figure 1. Wavelength division multiplexed passive optical network (WDM-PON) topologies with reflective semiconductor optical amplifier (RSOA) based colorless upstream (US) transmitters: (a) Amplified spontaneous emitter (ASE) spectrum-sliced transmitter: the US signal is directly modulated on to the ASE of the RSOA. The WDM filter at the remote node (RN) passes only the assigned sub band from each RSOA at the optical network units (ONUs); (b) Self-seeded RSOA-FCL (fiber cavity laser): a Fabry-Perot cavity indicated by the red dashed lines is formed between the RSOA and a mirror at the RN. The cavity is embedded in the distribution fiber (DF) of the PON; (c) Externally seeded RSOA: a laser at the central office (CO) seeds the RSOA at the ONU. The RSOA encodes the US signal onto the seed laser line. In all case, the ONUs are connected by the DF to the RN. The RN consists of the WDM filter and it is connected to the CO through the feeder fiber (FF). Figure 1. Wavelength division multiplexed passive optical network (WDM-PON) topologies with reﬂective semiconductor optical ampliﬁer (RSOA) based colorless upstream (US) transmitters: (a) Ampliﬁed spontaneous emitter (ASE) spectrum-sliced transmitter: the US signal is directly modulated on to the ASE of the RSOA. The WDM ﬁlter at the remote node (RN) passes only the assigned sub band from each RSOA at the optical network units (ONUs); (b) Self-seeded RSOA-FCL (ﬁber cavity laser): a Fabry-Perot cavity indicated by the red dashed lines is formed between the RSOA and a mirror at the RN. The cavity is embedded in the distribution ﬁber (DF) of the PON; (c) Externally seeded RSOA: a laser at the central ofﬁce (CO) seeds the RSOA at the ONU. The RSOA encodes the US signal onto the seed laser line. In all case, the ONUs are connected by the DF to the RN. The RN consists of the WDM ﬁlter and it is connected to the CO through the feeder ﬁber (FF). Figure 1. 2. DMT Signal Generation and Reception g p DMT is a multi-carrier transmission t In our experiments, the modulation schemes ranged from binary phase shift keying (BPSK with 1 bit/symbol) to 64QAM with 6 bit/symbol. The constellation diagrams of the modulation formats are shown in Figure 3. After bit mapping, the power of each subcarrier is optimized in a power-loading stage depending on the SNR profile, see Figure 2. This is done such that all subcarriers will end up with a relatively similar bit error ratio (BER) performance even if they have different modulation formats. The adaptive bit-and power-loading algorithm is based on Chow’s method discussed in [38]. In a next step, an inverse fast Fourier transform (IFFT) with Hermitian symmetry is performed to obtain the real valued DMT symbols. A cyclic prefix is added to each DMT symbol before being serialized and loaded into an arbitrary waveform generator (AWG). The AWG converts the DMT signal to the analog domain before it is fed to an RF amplifier and encoded to the optical domain by means of an RSOA. Figure 2. Discrete multi-tone (DMT) signal generation and reception flowchart. A pseudorandom bit sequence (PRBS) is generated, which is then rearranged into a parallel bit-stream. Subsequently, adaptive bit mapping and power-loading are implemented. Inverse fast Fourier transform (IFFT) is then used to encode the quadrature amplitude modulation (QAM) symbols on the DMT subcarriers. Cyclic prefix (CP) is added on the DMT symbols before the signal is converted back to serial. The serial data is loaded onto an arbitrary waveform generator (AWG) which converts the digital signal to analog using an inbuilt digital to analog converter (DAC). On the receiver side the signal is detected by a photodiode and sampled by a real-time oscilloscope (RTO).Next, timing synchronization is performed to recover the DMT clock. Then the serial signal is converted into a parallel data stream. After CP removal, FFT is used to decode the DMT subcarriers. Frequency and phase equalization of the received QAM symbols is performed. Here, we measure the error vector magnitude (EVM) and signal-to-noise ratio (SNR). Then, the symbols on each subcarrier are demapped to recover the transmitted PRBS for bit error ratio (BER) measurements. Figure 2. Discrete multi-tone (DMT) signal generation and reception ﬂowchart. A pseudorandom bit sequence (PRBS) is generated, which is then rearranged into a parallel bit-stream. Subsequently, adaptive bit mapping and power-loading are implemented. 2. DMT Signal Generation and Reception g p DMT is a multi-carrier transmission t DMT is a multi-carrier transmission technique with real-valued orthogonal subcarriers. The signal processing ﬂow charts for a DMT transmitter and receiver are given in Figure 2. A pseudorandom bit sequence (PRBS) is generated to test the transmitter. The bits are then rearranged from serial into parallel according to the number of subcarriers and the information content in the respective subcarriers. Each parallel bit stream is then mapped onto its individual constellation (e.g., quadrature amplitude modulation—QAM). DMT gives us the ﬂexibility to encode a different modulation format onto every subcarrier. Therefore, depending on the available SNR at each subcarrier the number of bits per symbol can be adapted to achieve the highest possible throughput, a feature called bit-loading. In our experiments, the modulation schemes ranged from binary phase shift keying (BPSK with 1 bit/symbol) to 64QAM with 6 bit/symbol. The constellation diagrams of the modulation formats are shown in Figure 3. After bit mapping, the power of each subcarrier is optimized in a power-loading stage depending on the SNR proﬁle, see Figure 2. This is done such that all subcarriers will end up with a relatively similar bit error ratio (BER) performance even if they have different modulation formats. The adaptive bit-and power-loading algorithm is based on Chow’s method discussed in [38]. In a next step, an inverse fast Fourier transform (IFFT) with Hermitian symmetry is performed to obtain the real valued DMT symbols. A cyclic preﬁx is added to each DMT symbol before being serialized and loaded into an arbitrary waveform generator (AWG). The AWG converts the DMT signal to the analog domain before it is fed to an RF ampliﬁer and encoded to the optical domain by means of an RSOA. q g signal processing flow charts for a DMT transmitter and receiver are given in Figure 2. A pseudorandom bit sequence (PRBS) is generated to test the transmitter. The bits are then rearranged from serial into parallel according to the number of subcarriers and the information content in the respective subcarriers. Each parallel bit stream is then mapped onto its individual constellation (e.g., quadrature amplitude modulation—QAM). DMT gives us the flexibility to encode a different modulation format onto every subcarrier. Therefore, depending on the available SNR at each subcarrier the number of bits per symbol can be adapted to achieve the highest possible throughput, a feature called bit-loading. 2. DMT Signal Generation and Reception g p DMT is a multi-carrier transmission t Inverse fast Fourier transform (IFFT) is then used to encode the quadrature amplitude modulation (QAM) symbols on the DMT subcarriers. Cyclic preﬁx (CP) is added on the DMT symbols before the signal is converted back to serial. The serial data is loaded onto an arbitrary waveform generator (AWG) which converts the digital signal to analog using an inbuilt digital to analog converter (DAC). On the receiver side the signal is detected by a photodiode and sampled by a real-time oscilloscope (RTO).Next, timing synchronization is performed to recover the DMT clock. Then the serial signal is converted into a parallel data stream. After CP removal, FFT is used to decode the DMT subcarriers. Frequency and phase equalization of the received QAM symbols is performed. Here, we measure the error vector magnitude (EVM) and signal-to-noise ratio (SNR). Then, the symbols on each subcarrier are demapped to recover the transmitted PRBS for bit error ratio (BER) measurements. Figure 2. Discrete multi-tone (DMT) signal generation and reception flowchart. A pseudorandom bit sequence (PRBS) is generated, which is then rearranged into a parallel bit-stream. Subsequently, adaptive bit mapping and power-loading are implemented. Inverse fast Fourier transform (IFFT) is then used to encode the quadrature amplitude modulation (QAM) symbols on the DMT subcarriers. Cyclic prefix (CP) is added on the DMT symbols before the signal is converted back to serial. The serial data is loaded onto an arbitrary waveform generator (AWG) which converts the digital signal to analog using an inbuilt digital to analog converter (DAC). On the receiver side the signal is detected by a photodiode and sampled by a real-time oscilloscope (RTO).Next, timing synchronization is performed to recover the DMT clock. Then the serial signal is converted into a parallel data stream. After CP removal, FFT is used to decode the DMT subcarriers. Frequency and phase equalization of the received QAM symbols is performed. Here, we measure the error vector magnitude (EVM) and signal-to-noise ratio (SNR). Then, the symbols on each subcarrier are demapped to recover the transmitted PRBS for bit error ratio (BER) measurements. Figure 2. Discrete multi-tone (DMT) signal generation and reception ﬂowchart. A pseudorandom bit sequence (PRBS) is generated, which is then rearranged into a parallel bit-stream. Subsequently, adaptive bit mapping and power-loading are implemented. Inverse fast Fourier transform (IFFT) is then used to encode the quadrature amplitude modulation (QAM) symbols on the DMT subcarriers. 1. Introduction The results are an expansion of our recent work [37], where only the maximized bit rates are presented. In this work, the SNR offered by each sources has been measured and compared. It is found that the SNR improves by more than 5 dB when switching from an ASE spectrum-sliced scheme to a self-seeded RSOA-FCL, and it improves by another ~5 dB when switching from a self-seeded RSOA-FCL to an externally seeded RSOA. Finally, transmission experiments have been performed. We demonstrate 6.25, 20.1, and 30.7 Gbit/s in back-to-back (BtB) and 4.17, 10.1 and 24.5 Gbit/s over 25 km NZDSF (non-zero dispersion shifted ﬁber) for the ASE spectrum-sliced, RSOA-FCL and externally seeded RSOA sources, respectively. No offset ﬁltering was needed in any of these experiments [14,25]. The organization of the paper is as follows First we discuss the steps for generating and analyzing y p y g y experiments [14,25]. The organization of the paper is as follows. First, we discuss the steps for generating and analyzing a DMT signal. In Section 3, we present the experimental setups. In Section 4, we measure The organization of the paper is as follows. First, we discuss the steps for generating and analyzing a DMT signal. In Section 3, we present the experimental setups. In Section 4, we measure the SNR of the RSOA-FCL by varying WDM ﬁlter bandwidth, cavity length and bias current. We compare 4 of 17 4 of 17 Appl. Sci. 2017, 7, 999 Appl. Sci. 2017, 7, 999 these results with the optimum SNR of the ASE spectrum-sliced and externally seeded RSOA schemes. Finally we present the results of bit- and power-loaded DMT transmission experiments. p p p y RSOA schemes. Finally we present the results of bit- and power-loaded DMT transmission experiments. 2 DMT Signal Generation and Reception 2. DMT Signal Generation and Reception g p DMT is a multi-carrier transmission t Cyclic preﬁx (CP) is added on the DMT symbols before the signal is converted back to serial. The serial data is loaded onto an arbitrary waveform generator (AWG) which converts the digital signal to analog using an inbuilt digital to analog converter (DAC). On the receiver side the signal is detected by a photodiode and sampled by a real-time oscilloscope (RTO).Next, timing synchronization is performed to recover the DMT clock. Then the serial signal is converted into a parallel data stream. After CP removal, FFT is used to decode the DMT subcarriers. Frequency and phase equalization of the received QAM symbols is performed. Here, we measure the error vector magnitude (EVM) and signal-to-noise ratio (SNR). Then, the symbols on each subcarrier are demapped to recover the transmitted PRBS for bit error ratio (BER) measurements. 5 of 17 red by Appl. Sci. 2017, 7, 999 transmitted and r i h Figure 3. Constellation diagrams of modulation formats used for the bit- and power-loaded DMT signal generation. Binary phase shift keying (BPSK) encodes 1 bit/symbol. Quadrature phase shift keying (QPSK) encodes 2 bit/symbol. The quadrature amplitude modulation (QAM) formats: 8QAM, 16QAM, 32QAM, and 64QAM encode 3, 4, 5 and 6 bit/symbol, respectively. Figure 3. Constellation diagrams of modulation formats used for the bit- and power-loaded DMT signal generation. Binary phase shift keying (BPSK) encodes 1 bit/symbol. Quadrature phase shift keying (QPSK) encodes 2 bit/symbol. The quadrature amplitude modulation (QAM) formats: 8QAM, 16QAM, 32QAM, and 64QAM encode 3, 4, 5 and 6 bit/symbol, respectively. Figure 3. Constellation diagrams of modulation formats used for the bit- and power-loaded DMT signal generation. Binary phase shift keying (BPSK) encodes 1 bit/symbol. Quadrature phase shift keying (QPSK) encodes 2 bit/symbol. The quadrature amplitude modulation (QAM) formats: 8QAM, 16QAM, 32QAM, and 64QAM encode 3, 4, 5 and 6 bit/symbol, respectively. Figure 3. Constellation diagrams of modulation formats used for the bit- and power-loaded DMT signal generation. Binary phase shift keying (BPSK) encodes 1 bit/symbol. Quadrature phase shift keying (QPSK) encodes 2 bit/symbol. The quadrature amplitude modulation (QAM) formats: 8QAM, 16QAM, 32QAM, and 64QAM encode 3, 4, 5 and 6 bit/symbol, respectively. 3. Experimental Setups In our experiments, we investigated the three colorless US transmitters in WDM-PON scenarios, see Figure 4. For all experiments, the transmitted DMT signal is generated offline with MATLAB and converted to an analog waveform by an AWG (Keysight M8195A—60 GSa/s, Santa Rosa, CA, USA). Figure 4a shows the ROSA in the ONU is mo 3. Experimental Setups power of the RSOA. At the remote note, an OBPF (tuned to the emission center of the RSOA, which is at 1537.92 nm) band limits the modulated ASE-signal to the assigned wavelength channel. The filtered signal is then sent to the CO. The filter bandwidth has been chosen to be identical to the filter bandwidth of the other schemes. Figure 4b shows the setup for the self-seeded RSOA-FCL. Here, a Faraday rotator (FR) is placed at ONU after the RSOA. In addition, a Faraday rotator mirror (FRM) is placed at the RN after a coupler behind the WDM filter. The FRM and the RSOA reflective facet form the Fabry-Perot resonator cavity embedded in the DF. In case of RSOA with high polarization dependent gain In our experiments, we investigated the three colorless US transmitters in WDM-PON scenarios, see Figure 4. For all experiments, the transmitted DMT signal is generated ofﬂine with MATLAB and converted to an analog waveform by an AWG (Keysight M8195A—60 GSa/s, Santa Rosa, CA, USA). The analog signal is ampliﬁed and a DC bias current is added before being fed to an RSOA. In all cases, a 1 mm long InGaAsP quantum well C-band RSOA (provided by III-V labs, 10G-RSOA-11-S, Palaiseau, France) is placed at the ONU. The distribution ﬁber (DF) links the ONU to the RN. At the RN, a ﬂat top optical band-pass ﬁlter (OBPF) emulates the WDM ﬁlter. RN and CO are connected by the feeder ﬁber (FF). (HPDG), the FR and FRM cancel out the birefringence effect in the cavity and maintain a constant polarization at the RSOA input [42]. The coupler at the RN has a coupling ratio of 90/10, where 90% The three implementations of the RSOA based transmission links are shown in Figure 4 and described in more detailed now. of the power is kept in the cavity and 10% is transmitted to the CO. The large amount of the power kept in the cavity is required to overcome cavity losses and thereby to achieve a net gain and with this lasing. In addition, the high power in the cavity maintains the RSOA in the nonlinear operation region for better noise and residual modulation suppression [17,43]. Figure 4c shows the experimental setup for the externally seeded RSOA scheme. 2. DMT Signal Generation and Reception g p DMT is a multi-carrier transmission t The analog signal is amplified and a DC bias current is added before being fed to an RSOA. In all cases, a 1 mm long InGaAsP quantum well C-band RSOA (provided by III-V labs, 10G-RSOA-11-S, Palaiseau, France) is placed at the ONU. The distribution fiber (DF) links the ONU to the RN. At the RN, a flat top optical band-pass filter (OBPF) emulates the WDM filter. RN and CO are connected by the feeder fiber (FF). The three implementations of the RSOA based transmission links are shown in Figure 4 and described in more detailed now. In the receiver, a direct detection scheme is used to detect the optical signals. The electrical signals are then fed into a real-time oscilloscope (RTO). The RTO effectively digitizes the signal with a resolution of ~5.8 bits. In a ﬁrst step, timing synchronization is performed using the non-data aided DMT timing estimation technique discussed in [39]. Subsequently, the signal is parallelized and the cyclic preﬁx is removed.The subcarriers within the electrical signal are then derived by the fast Fourier transform (FFT). To properly decode the bits within a subcarrier, blind phase and frequency offset estimation methods are applied on every subcarrier [40]. The error vector magnitude (EVM) of the QAM symbols on each subcarrier is calculated and converted into SNR [41]. Then, the QAM symbols are de-mapped to extract the received bit pattern. The BER is measured by comparing the transmitted and received bit patterns on each subcarrier. The overall BER is also measured by comparing the total transmitted and received bits. Figure 4a shows the ROSA in the ONU is mo 3. Experimental Setups At the optical network units (ONUs), the upstream signal from the arbitrary waveform generator (AWG) is amplified and added to a bias current to drive the RSOA. The ONU and the remote node (RN) are connected by the distribution fiber (DF). At the RN, an optical bandpass filter (OBPF) mimics the WDM filter. In the case of the RSOA-FCL in (b), the Faraday rotator mirror (FRM) is placed behind the filter and a coupler to form the Fabrey-Perot cavity, shown by the red dashed line. For the externally-seeded RSOA in (c), the seeding cw laser is place at the central office (CO). On the receiver side at the CO, pre-amplification and direct detection are performed. A real-time oscilloscope (RTO) records the electrical signal after the photodiode. An optical spectrum analyzer (OSA) is used to monitor the optical signal at the CO. EDFA: Erbium doped fiber amplifier, VOA: variable optical attenuator. I the th ee he e e i le e t o ly a i le o ti al ha el ea u e e t due to la k of Figure 4. Experimental setups of three RSOA based colorless transmitter schemes in a WDM-PON scenario: (a) ASE spectrum-sliced, (b) self-seeded RSOA-FCL, and (c) externally seeded RSOA. At the optical network units (ONUs), the upstream signal from the arbitrary waveform generator (AWG) is ampliﬁed and added to a bias current to drive the RSOA. The ONU and the remote node (RN) are connected by the distribution ﬁber (DF). At the RN, an optical bandpass ﬁlter (OBPF) mimics the WDM ﬁlter. In the case of the RSOA-FCL in (b), the Faraday rotator mirror (FRM) is placed behind the ﬁlter and a coupler to form the Fabrey-Perot cavity, shown by the red dashed line. For the externally-seeded RSOA in (c), the seeding cw laser is place at the central ofﬁce (CO). On the receiver side at the CO, pre-ampliﬁcation and direct detection are performed. A real-time oscilloscope (RTO) records the electrical signal after the photodiode. An optical spectrum analyzer (OSA) is used to monitor the optical signal at the CO. EDFA: Erbium doped ﬁber ampliﬁer, VOA: variable optical attenuator. equipment, however more than 16 channels have been demonstrated [2,21]. In all cases, the link budget between the ONU and the CO is ~12 dB (i.e., combination of all insertion losses of components in the link between ONU and CO). Figure 4a shows the ROSA in the ONU is mo 3. Experimental Setups Here, a laser source (emitting at 1537.92 nm, similar to the other setups) at the CO seeds the RSOA at the ONU Figure 4a shows the setup for the ASE spectrum-sliced scheme. In this scheme, the current of an ROSA in the ONU is modulated with the data. This encodes the information onto the ASE output power of the RSOA. At the remote note, an OBPF (tuned to the emission center of the RSOA, which is at 1537.92 nm) band limits the modulated ASE-signal to the assigned wavelength channel. The ﬁltered signal is then sent to the CO. The ﬁlter bandwidth has been chosen to be identical to the ﬁlter bandwidth of the other schemes. Figure 4b shows the setup for the self-seeded RSOA-FCL. Here, a Faraday rotator (FR) is placed at ONU after the RSOA. In addition, a Faraday rotator mirror (FRM) is placed at the RN after a coupler behind the WDM ﬁlter. The FRM and the RSOA reﬂective facet form the Fabry-Perot resonator cavity embedded in the DF. In case of RSOA with high polarization dependent gain (HPDG), the FR and FRM cancel out the birefringence effect in the cavity and maintain a constant polarization at the RSOA input [42]. The coupler at the RN has a coupling ratio of 90/10, where 90% of the power is kept in the cavity and 10% is transmitted to the CO. The large amount of the power kept in the cavity is required to overcome cavity losses and thereby to achieve a net gain and with this lasing. In addition, the high power in the cavity maintains the RSOA in the nonlinear operation region for better noise and residual modulation suppression [17,43]. 6 of 17 Appl. Sci. 2017, 7, 999 Figure 4c shows the experimental setup for the externally seeded RSOA scheme. Here, a laser source (emitting at 1537.92 nm, similar to the other setups) at the CO seeds the RSOA at the ONU through the ﬁber link. The seeding power entering the RSOA has been optimized for the highest SNR. The RSOA now only serves as a modulator and ampliﬁer for the respective laser wavelength. Appl. Sci. 2017, 7, 999 6 of 17 through the fiber link. The seeding power entering the RSOA has been optimized for the highest SNR. The RSOA now only serves as a modulator and amplifier for the respective laser wavelength. Figure 4. Figure 4a shows the ROSA in the ONU is mo 3. Experimental Setups At the CO, a direct detection receiver detects the signals from the three transmitters. Here, a 90/10 coupler sends 10% of the power to an optical spectrum analyzer (OSA) to monitor the signal. A variable optical attenuator (VOA) is used to control the power entering the preamplifier Erbium doped fiber amplifier (EDFA). A pre-amplified direct detection receiver is used here to guarantee that the signal quality measured here is as much as possible independent of the receiver scheme. A 5 nm flat top OBPF suppresses the out of band ASE noise from the EDFA. The optical signal is finally detected by a photodiode (PIN, responsivity 0.6 A/W), which is directly connected to a RTO (DSOX96204Q—20 GHz bandwidth and 40 GSa/s). The oscilloscope records the signal for offline post-processing with MATLAB as outlined in Figure 2. 4. SNR Characterization In this section we present the measured SNRs versus subcarrier frequency offered by the three In the three schemes, we implement only a single optical channel measurement due to lack of equipment, however more than 16 channels have been demonstrated [2,21]. In all cases, the link budget between the ONU and the CO is ~12 dB (i.e., combination of all insertion losses of components in the link between ONU and CO). At the CO, a direct detection receiver detects the signals from the three transmitters. Here, a 90/10 coupler sends 10% of the power to an optical spectrum analyzer (OSA) to monitor the signal. A variable optical attenuator (VOA) is used to control the power entering the preampliﬁer Erbium doped ﬁber ampliﬁer (EDFA). A pre-ampliﬁed direct detection receiver is used here to guarantee that the signal quality measured here is as much as possible independent of the receiver scheme. A 5 nm ﬂat top OBPF suppresses the out of band ASE noise from the EDFA. The optical signal is ﬁnally detected by a photodiode (PIN, responsivity 0.6 A/W), which is directly connected to a RTO (DSOX96204Q—20 GHz bandwidth and 40 GSa/s). The oscilloscope records the signal for ofﬂine post-processing with MATLAB as outlined in Figure 2. Figure 4a shows the ROSA in the ONU is mo 3. Experimental Setups Experimental setups of three RSOA based colorless transmitter schemes in a WDM-PON scenario: (a) ASE spectrum-sliced, (b) self-seeded RSOA-FCL, and (c) externally seeded RSOA. At the optical network units (ONUs), the upstream signal from the arbitrary waveform generator (AWG) is amplified and added to a bias current to drive the RSOA. The ONU and the remote node (RN) are connected by the distribution fiber (DF). At the RN, an optical bandpass filter (OBPF) mimics the WDM filter. In the case of the RSOA-FCL in (b), the Faraday rotator mirror (FRM) is placed behind the filter and a coupler to form the Fabrey-Perot cavity, shown by the red dashed line. For the externally-seeded RSOA in (c), the seeding cw laser is place at the central office (CO). On the receiver side at the CO, pre-amplification and direct detection are performed. A real-time oscilloscope (RTO) records the electrical signal after the photodiode. An optical spectrum analyzer (OSA) is used to monitor the optical signal at the CO. EDFA: Erbium doped fiber amplifier, VOA: variable optical attenuator. In the three schemes, we implement only a single optical channel measurement due to lack of Figure 4. Experimental setups of three RSOA based colorless transmitter schemes in a WDM-PON scenario: (a) ASE spectrum-sliced, (b) self-seeded RSOA-FCL, and (c) externally seeded RSOA. At the optical network units (ONUs), the upstream signal from the arbitrary waveform generator (AWG) is ampliﬁed and added to a bias current to drive the RSOA. The ONU and the remote node (RN) are connected by the distribution ﬁber (DF). At the RN, an optical bandpass ﬁlter (OBPF) mimics the WDM ﬁlter. In the case of the RSOA-FCL in (b), the Faraday rotator mirror (FRM) is placed behind the ﬁlter and a coupler to form the Fabrey-Perot cavity, shown by the red dashed line. For the externally-seeded RSOA in (c), the seeding cw laser is place at the central ofﬁce (CO). On the receiver side at the CO, pre-ampliﬁcation and direct detection are performed. A real-time oscilloscope (RTO) records the electrical signal after the photodiode. An optical spectrum analyzer (OSA) is used to monitor the optical signal at the CO. EDFA: Erbium doped ﬁber ampliﬁer, VOA: variable optical attenuator. Figure 4. Experimental setups of three RSOA based colorless transmitter schemes in a WDM-PON scenario: (a) ASE spectrum-sliced, (b) self-seeded RSOA-FCL, and (c) externally seeded RSOA. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance Here, we perform three studies to analyze the performance of the RSOA-FCL: First, we investigate the impact of the filter bandwidth in the cavity. Second, we study the SNR dependence on cavity length and bias current. Last, we explore the transmission performance for different cavity lengths at optimized operation conditions. In all cases, we use the SNR over the modulation frequency as a measure for the link performance and for comparing parameters. The SNR over the modulation frequency is measured according to the following procedure We Here, we perform three studies to analyze the performance of the RSOA-FCL: First, we investigate the impact of the ﬁlter bandwidth in the cavity. Second, we study the SNR dependence on cavity length and bias current. Last, we explore the transmission performance for different cavity lengths at optimized operation conditions. In all cases, we use the SNR over the modulation frequency as a measure for the link performance and for comparing parameters. The SNR over the modulation frequency is measured according to the following procedure. We transmit DMT signals and evaluate the SNR on each subcarrier at the receiver. Here, the modulation format and power of each subcarrier is identical. We generate a DMT signal having 256 subcarriers with QPSK as modulation format and equal electrical power for all subcarriers. A PRBS 11 is used as the transmitted bit sequence. The total symbol rate is 10 GBd. The IFFT size is 512 and a 2.5% cyclic prefix is added to each DMT symbol. The signal is generated offline with MATLAB An AWG generates the analog signal with 0 5 The SNR over the modulation frequency is measured according to the following procedure. We transmit DMT signals and evaluate the SNR on each subcarrier at the receiver. Here, the modulation format and power of each subcarrier is identical. We generate a DMT signal having 256 subcarriers with QPSK as modulation format and equal electrical power for all subcarriers. A PRBS 11 is used as the transmitted bit sequence. The total symbol rate is 10 GBd. The IFFT size is 512 and a 2.5% cyclic preﬁx is added to each DMT symbol. The signal is generated offline with MATLAB. An AWG generates the analog signal with 0.5 Vpp (peak-to-peak voltage). The output of the RF amplifier was optimized to ~3.5 Vpp. transmitter schemes. Be ea u i the ele t i a 4. SNR Characterization measuring the electrical modulation bandwidth. In a second step, we then focus on the self seeded RSOA-FCL. We study the impact of the WDM filter bandwidth in the RN, the fiber cavity length and In this section, we present the measured SNRs versus subcarrier frequency offered by the three transmitter schemes. Before presenting the results, we study the performance of the RSOA itself by 7 of 17 Appl. Sci. 2017, 7, 999 measuring the electrical modulation bandwidth. In a second step, we then focus on the self-seeded RSOA-FCL. We study the impact of the WDM ﬁlter bandwidth in the RN, the ﬁber cavity length and the bias current on the SNR offered by the RSOA-FCL. Once the optimum operation parameters and limitations of the RSOA-FCL have been understood, we adjust the operation points of the ASE spectrum-sliced and the externally seeded RSOA. So for instance we set the WDM ﬁlter bandwidth of the ASE spectrum-sliced source to be identical with the RSOA-FCL. The input power for the externally seeded transmitters is also set for maximum performance. Here we only give the operation points that have been selected while one can ﬁnd more details on ﬁnding the ideal operation points of an ASE spectrum-sliced transmitter in [44,45] and the externally seeded transmitter [25,46]. Appl. Sci. 2017, 7, 999 7 of 17 the bias current on the SNR offered by the RSOA-FCL. Once the optimum operation parameters and limitations of the RSOA-FCL have been understood, we adjust the operation points of the ASE spectrum-sliced and the externally seeded RSOA. So for instance we set the WDM filter bandwidth of the ASE spectrum-sliced source to be identical with the RSOA-FCL. The input power for the externally seeded transmitters is also set for maximum performance. Here we only give the operation points that have been selected while one can find more details on finding the ideal operation points of an ASE spectrum-sliced transmitter in [44,45] and the externally seeded transmitter [25,46]. 4.1. Electrical Bandwidth of the RSOA 4.1. Electrical Bandwidth of the RSOA We start our investigation by measuring the small signal modulation frequency response of the RSOA itself. We chose the externally seeded conﬁguration to exclude the impact of dispersion and the external cavity. The frequency response for 50–120 mA bias current and at an input power of −3 dBm are plotted in Figure 5, which shows that the modulation response improves with higher bias current. At 100 mA, the RSOA exhibits −3 and −6 dB bandwidths of 2.4 and 4.55 GHz, respectively. We start our investigation by measuring the small signal modulation frequency response of the RSOA itself. We chose the externally seeded configuration to exclude the impact of dispersion and the external cavity. The frequency response for 50–120 mA bias current and at an input power of −3 dBm are plotted in Figure 5, which shows that the modulation response improves with higher bias current. At 100 mA, the RSOA exhibits −3 and −6 dB bandwidths of 2.4 and 4.55 GHz, respectively. Figure 5. Measured small signal modulation frequency response of an RSOA at 50–120 mA bias currents. The modulation bandwidth increases with bias current. At 100 mA, the RSOA has −3 and −6 dB bandwidths of 2.4 and 4.55 GHz. Figure 5. Measured small signal modulation frequency response of an RSOA at 50–120 mA bias currents. The modulation bandwidth increases with bias current. At 100 mA, the RSOA has −3 and −6 dB bandwidths of 2.4 and 4.55 GHz. Figure 5. Measured small signal modulation frequency response of an RSOA at 50–120 mA bias currents. The modulation bandwidth increases with bias current. At 100 mA, the RSOA has −3 and −6 dB bandwidths of 2.4 and 4.55 GHz. Figure 5. Measured small signal modulation frequency response of an RSOA at 50–120 mA bias currents. The modulation bandwidth increases with bias current. At 100 mA, the RSOA has −3 and −6 dB bandwidths of 2.4 and 4.55 GHz. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance A bias tee combines the amplified signal with the specific bias current to drive the RSOA. On the receiver side, an RTO records the signal detected by a photodetector. Subsequently, the signal is demodulated in an offline digital signal processing step The measured EVM of the subcarriers is used to calculate the The signal is generated ofﬂine with MATLAB. An AWG generates the analog signal with 0.5 Vpp (peak-to-peak voltage). The output of the RF ampliﬁer was optimized to ~3.5 Vpp. A bias tee combines the ampliﬁed signal with the speciﬁc bias current to drive the RSOA. On the receiver side, an RTO records the signal detected by a photodetector. Subsequently, the signal is demodulated in an ofﬂine Appl. Sci. 2017, 7, 999 8 of 17 digital signal processing step. The measured EVM of the subcarriers is used to calculate the SNR of each subcarrier. This way, we can determine the frequency resolved SNR of the transmission link across the whole modulation bandwidth. This allows us to determine the usable bandwidth of the transmission link. The useable bandwidth is the frequency band, within which the SNR is high enough to still allow for a BER below 10−3 with BPSK modulation. Theoretically, a minimum SNR of 6.7 dB is required. Appl. Sci. 2017, 7, 999 8 of 17 enough to still allow for a BER below 10−3 with BPSK modulation. Theoretically, a minimum SNR of 6 7 dB is required First, we investigated the impact of the WDM ﬁlter bandwidth in the RN. We compared OBPF bandwidths of 0.6 nm and 2 nm for three cavity lengths of 15 m, 115 m and 1 km standard single mode ﬁber (SSMF). The bias current was maintained at 100 mA. Figure 6 shows the SNR over the modulation frequency for both cases. The SNR is higher for the ﬁlter with wider bandwidth for all cavity lengths. For a 0.6 nm ﬁlter, the 15 and 115 m cavities have a peak SNRs of ~14 dB. The maximum SNR obtained for the 1 km cavity with 0.6 nm is only ~12 dB. The usable bandwidth of the 15, 115 m and 1 km cavities with 0.6 nm ﬁlter was ~4.6, 4.5 and 2.7 GHz. When a 2 nm ﬁlter has been used in the receiver one ﬁnds SNRs that are 5 dB higher. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance The usable bandwidth then increases by ~40% to ~6.7, ~6.4 and ~4.5 GHz. The results are summarized in Table 1. 6.7 dB is required. First, we investigated the impact of the WDM filter bandwidth in the RN. We compared OBPF bandwidths of 0.6 nm and 2 nm for three cavity lengths of 15 m, 115 m and 1 km standard single mode fiber (SSMF). The bias current was maintained at 100 mA. Figure 6 shows the SNR over the modulation frequency for both cases. The SNR is higher for the filter with wider bandwidth for all cavity lengths. For a 0.6 nm filter, the 15 and 115 m cavities have a peak SNRs of ~14 dB. The maximum SNR obtained for the 1 km cavity with 0.6 nm is only ~12 dB. The usable bandwidth of the 15, 115 m and 1 km cavities with 0.6 nm filter was ~4.6, 4.5 and 2.7 GHz. When a 2 nm filter has been used in the receiver one finds SNRs that are 5 dB higher. The usable bandwidth then increases by ~40% to ~6.7, ~6.4 and ~4.5 GHz. The results are summarized in Table 1. Figure 6. Measured SNR comparison of self-seeded RSOA-FCL transmitter with WDM filter bandwidths of 0.6 and 2 nm in RN. The dotted and solid lines show the measured SNR for the 0.6 and 2 nm filters, respectively. For both cases, three different cavity lengths, 15 m, 115 m and 1 km were used. 0.6 nm filter cavity has peak SNR which is ~5 dB lower than that of the 2 nm one. Figure 6. Measured SNR comparison of self-seeded RSOA-FCL transmitter with WDM ﬁlter bandwidths of 0.6 and 2 nm in RN. The dotted and solid lines show the measured SNR for the 0.6 and 2 nm ﬁlters, respectively. For both cases, three different cavity lengths, 15 m, 115 m and 1 km were used. 0.6 nm ﬁlter cavity has peak SNR which is ~5 dB lower than that of the 2 nm one. Figure 6. Measured SNR comparison of self-seeded RSOA-FCL transmitter with WDM filter bandwidths of 0.6 and 2 nm in RN. The dotted and solid lines show the measured SNR for the 0.6 and 2 nm filters, respectively. For both cases, three different cavity lengths, 15 m, 115 m and 1 km were used. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance For example, at 100 mA, the reflected power into the RSOA was ~−6 dBm and ~−2 dBm for the 0.6 and 2 nm filters, respectively. Thus, higher losses means that the RSOA is less saturated. Therefore, the RSOA will be unable to efficiently suppress the residual modulation and noise As a consequence the RIN will be higher and the SNR is degraded [47] The second Table 1. Summary of results to compare the peak signal-to-noise ratio (SNR) and usable SNR bandwidth for the RSOA-FCL (reﬂective semiconductor optical ampliﬁer ﬁber cavity laser) with 0.6 nm or 2 nm ﬁlter bandwidths. For both ﬁlters, the ﬁber cavity length varied between 15 m, 115 m and 1 km. OBPF: optical band-pass ﬁlter. explanation based on [48] is that in case of highly multimode sources, increasing the number of modes (in this case wider filter) will distribute the noise over the multiple modes thus lowering the noise spectral density and RIN. Subsequently, we continue the investigation with the 2 nm filter only. Table 1. Summary of results to compare the peak signal-to-noise ratio (SNR) and usable SNR bandwidth for the RSOA-FCL (reflective semiconductor optical amplifier fiber cavity laser) with 0.6 nm or 2 nm filter bandwidths. For both filters, the fiber cavity length varied between 15 m, 115 m and 1 km. OBPF: optical band-pass filter. RSOA-FCL Cavity Length OBPF −3 dB Bandwidth Peak SNR (dB) Usable SNR Bandwidth (GHz) 15 m 0.6 nm 14.3 4.6 2 nm 19.45 6.7 115 m 0.6 nm 14.1 4.5 2 nm 19.5 6.4 1 km 0.6 nm 11.9 2.7 2 nm 17.7 4.5 RSOA-FCL Cavity Length OBPF −3 dB Bandwidth Peak SNR (dB) Usable SNR Bandwidth (GHz) 15 m 0.6 nm 14.3 4.6 2 nm 19.45 6.7 115 m 0.6 nm 14.1 4.5 2 nm 19.5 6.4 1 km 0.6 nm 11.9 2.7 2 nm 17.7 4.5 One explanation for the SNR degradation with narrower ﬁlters would be that a narrower ﬁlter leads to higher losses [47]. For example, at 100 mA, the reﬂected power into the RSOA was ~−6 dBm and ~−2 dBm for the 0.6 and 2 nm ﬁlters, respectively. Thus, higher losses means that the RSOA is less saturated. Therefore, the RSOA will be unable to efﬁciently suppress the residual modulation and noise. As a consequence the RIN will be higher and the SNR is degraded [47]. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance 0.6 nm filter cavity has peak SNR which is ~5 dB lower than that of the 2 nm one. Figure 6. Measured SNR comparison of self-seeded RSOA-FCL transmitter with WDM ﬁlter bandwidths of 0.6 and 2 nm in RN. The dotted and solid lines show the measured SNR for the 0.6 and 2 nm ﬁlters, respectively. For both cases, three different cavity lengths, 15 m, 115 m and 1 km were used. 0.6 nm ﬁlter cavity has peak SNR which is ~5 dB lower than that of the 2 nm one. One explanation for the SNR degradation with narrower filters would be that a narrower filter leads to higher losses [47]. For example, at 100 mA, the reflected power into the RSOA was ~−6 dBm and ~−2 dBm for the 0.6 and 2 nm filters, respectively. Thus, higher losses means that the RSOA is less saturated. Therefore, the RSOA will be unable to efficiently suppress the residual modulation and noise. As a consequence the RIN will be higher and the SNR is degraded [47]. The second Table 1. Summary of results to compare the peak signal-to-noise ratio (SNR) and usable SNR bandwidth for the RSOA-FCL (reﬂective semiconductor optical ampliﬁer ﬁber cavity laser) with 0.6 nm or 2 nm ﬁlter bandwidths. For both ﬁlters, the ﬁber cavity length varied between 15 m, 115 m and 1 km. OBPF: optical band-pass ﬁlter. One explanation for the SNR degradation with narrower filters would be that a narrower filter leads to higher losses [47]. For example, at 100 mA, the reflected power into the RSOA was ~−6 dBm and ~−2 dBm for the 0.6 and 2 nm filters, respectively. Thus, higher losses means that the RSOA is less saturated. Therefore, the RSOA will be unable to efficiently suppress the residual modulation and noise. As a consequence the RIN will be higher and the SNR is degraded [47]. The second Table 1. Summary of results to compare the peak signal-to-noise ratio (SNR) and usable SNR bandwidth for the RSOA-FCL (reﬂective semiconductor optical ampliﬁer ﬁber cavity laser) with 0.6 nm or 2 nm ﬁlter bandwidths. For both ﬁlters, the ﬁber cavity length varied between 15 m, 115 m and 1 km. OBPF: optical band-pass ﬁlter. One explanation for the SNR degradation with narrower filters would be that a narrower filter leads to higher losses [47]. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance The similar performance originates from negligible chromatic dispersion for the two lengths. In both cases, the frequency dependent SNR increases with the bias current, which is related to the improved RSOA frequency response for higher bias currents as shown in Figure 5. The SNR degradation for higher modulation frequencies is due to the bandwidth limitations of the RSOA as discussed in Figure 5. At 110 mA, both achieve maximum SNR of ~19 dB. The usable bandwidth where the SNR is still sufficiently high to receive a BPSK signal is ~6.7 and ~6.4 GHz for cavity lengths of 15 m and 115 m, respectively. Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). In Figure 7c e ha e plotted the SNR for the longer 1 km SSMF ca ity It is ay lo er than those Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance The second explanation based on [48] is that in case of highly multimode sources, increasing the number of modes (in this case Appl. Sci. 2017, 7, 999 9 of 17 wider ﬁlter) will distribute the noise over the multiple modes thus lowering the noise spectral density and RIN. Subsequently, we continue the investigation with the 2 nm ﬁlter only. Appl Sci 2017 7 999 9 of 17 Next, we further investigated the impact of the distribution ﬁber cavity length and the bias current on the SNR of the self-seeded RSOA-FCL. We considered cavity lengths with 15 m, 115 m and 1 km SSMF to mimic WDM-PONs with distribution ﬁbers in the range of tens of meters up to a few kilometers, for example in FTTA [49]. The RSOA bias current was set at 80, 100, 110 and 120 mA. Figure 7a,b show similar performance for cavities with 15 m and 115 m length. The insets in Figure 7a,b which are the transmitted (black curve) and the received (red curve) DMT electrical spectrums also show similar characteristics. The similar performance originates from negligible chromatic dispersion for the two lengths. In both cases, the frequency dependent SNR increases with the bias current, which is related to the improved RSOA frequency response for higher bias currents as shown in Figure 5. The SNR degradation for higher modulation frequencies is due to the bandwidth limitations of the RSOA as discussed in Figure 5. At 110 mA, both achieve maximum SNR of ~19 dB. The usable bandwidth where the SNR is still sufﬁciently high to receive a BPSK signal is ~6.7 and ~6.4 GHz for cavity lengths of 15 m and 115 m, respectively. Appl. Sci. 2017, 7, 999 9 of 17 Next, we further investigated the impact of the distribution fiber cavity length and the bias current on the SNR of the self-seeded RSOA-FCL. We considered cavity lengths with 15 m, 115 m and 1 km SSMF to mimic WDM-PONs with distribution fibers in the range of tens of meters up to a few kilometers, for example in FTTA [49]. The RSOA bias current was set at 80, 100, 110 and 120 mA. Figure 7a,b show similar performance for cavities with 15 m and 115 m length. The insets in Figure 7a,b which are the transmitted (black curve) and the received (red curve) DMT electrical spectrums also show similar characteristics. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). in Figure 7a,b. This can be understood by the fact that total accumulated chromatic dispersion is larger. Since the RSOA-FCL is highly multi-mode, chromatic dispersion leads to considerable mode partition noise. This noise increases the RIN [50,51], causing a SNR degradation. A bit higher loss of the 1 km fiber cavity makes only a minor contribution to the SNR increase. At 110 mA, the maximum SNR is ~17 dB only and the usable bandwidth is now ~4.5 GHz. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, d A b f l h f b d k h l d Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). Figure 7. Measured SNR versus subcarrier frequency of self-seeded RSOA-FCLs operated at 80, 100, 110 and 120 mA bias currents for cavity lengths of (a) 15 m, (b) 115 m and (c) 1 km. The plots indicate that the SNR bandwidth generally increases with the bias current. The 1 km cavity shown in (c) has a reduced SNR peak and bandwidth when compared to the shorter cavities. The dashed line indicates the minimum SNR of 6.7 dB required to transmit BPSK, thus, it determines the usable bandwidth. In each plot, the insets show a sample electrical domain DMT spectra. The black and the red curves show the transmitted and received DMT electrical spectrum, respectively, at 110 mA. The received DMT spectrum for the 1 km cavity RSOA-FCL shown in plot (c) inset has a higher noise level compared to that of the 15 and 115 m cavities spectra shown in insets of (a,b). Figure 7. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance In contrast to the RSOA-FCL with shorter cavity lengths, the SNR in the 1 km cavity is not increasing with bias current. We expect that this originates from the strong chirp during direct modulation at higher bias currents. The chirp causes spectral broadening which increases chromatic dispersion penalty. This induces an additional In Figure 7c we have plotted the SNR for the longer 1 km SSMF cavity. It is way lower than those in Figure 7a,b. This can be understood by the fact that total accumulated chromatic dispersion is larger. Since the RSOA-FCL is highly multi-mode, chromatic dispersion leads to considerable mode partition noise. This noise increases the RIN [50,51], causing a SNR degradation. A bit higher loss of the 1 km ﬁber cavity makes only a minor contribution to the SNR increase. At 110 mA, the maximum SNR is ~17 dB only and the usable bandwidth is now ~4.5 GHz. In contrast to the RSOA-FCL with shorter cavity lengths, the SNR in the 1 km cavity is not increasing with bias current. We expect that this Appl. Sci. 2017, 7, 999 10 of 17 originates from the strong chirp during direct modulation at higher bias currents. The chirp causes spectral broadening which increases chromatic dispersion penalty. This induces an additional SNR degradation for higher bias currents [52]. This is supported by the large noise level in the received electrical DMT spectrum of the 1 km RSOA-FCL that can be seen in the inset of Figure 7c (red curve). However, the negative impact of the dispersive cavity on the SNR can be avoided by using O-band RSOAs. In the O-band where chromatic dispersion is minimum, RSOA-FCL cavity could be extended beyond 1 km [22,53]. Appl. Sci. 2017, 7, 999 10 of 17 SNR degradation for higher bias currents [52]. This is supported by the large noise level in the received electrical DMT spectrum of the 1 km RSOA-FCL that can be seen in the inset of Figure 7c (red curve). However, the negative impact of the dispersive cavity on the SNR can be avoided by using O-band RSOAs. In the O-band where chromatic dispersion is minimum, RSOA-FCL cavity could be extended beyond 1 km [22,53]. y Finally, we investigated the SNR after transmission over 25 km NZDSF (chromatic dispersion: ~5 ps/nm·km). 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance This ﬁber is used to emulate a low dispersion PON ﬁber—similar to what one might expect if the system were operated in the O-Band. The probed SNR for the three cavity lengths for both a BtB and a 25 km transmission are plotted in Figure 8. When comparing the BtB and 25 km SNRs, the maximum SNR values for the respective cavity lengths did not show major change. Rather, the usable SNR bandwidth was reduced to ~3.3, 3.2 and 2.8 GHz for the 15 m, 115 m and 1 km cavities, respectively. The results are summarized in Table 2. This shows a 40–50% bandwidth reduction compared to the BtB case. The ﬁnite signal bandwidth of the directly modulated RSOA results in chirp after propagation that induces power fading and bandwidth reduction. In addition, such highly multi-mode source suffers from mode partition noise induced by the NZDSF dispersion which increases the RIN. y [ , ] Finally, we investigated the SNR after transmission over 25 km NZDSF (chromatic dispersion: ~5 ps/nm·km). This fiber is used to emulate a low dispersion PON fiber—similar to what one might expect if the system were operated in the O-Band. The probed SNR for the three cavity lengths for both a BtB and a 25 km transmission are plotted in Figure 8. When comparing the BtB and 25 km SNRs, the maximum SNR values for the respective cavity lengths did not show major change. Rather, the usable SNR bandwidth was reduced to ~3.3, 3.2 and 2.8 GHz for the 15 m, 115 m and 1 km cavities, respectively. The results are summarized in Table 2. This shows a 40–50% bandwidth reduction compared to the BtB case. The finite signal bandwidth of the directly modulated RSOA results in chirp after propagation that induces power fading and bandwidth reduction. In addition, such highly multi-mode source suffers from mode partition noise induced by the NZDSF dispersion which increases the RIN. Figure 8. Measured SNR to compare the impact of transmission fiber on the performance of an RSOA- FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non- zero dispersion shifted fiber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km fiber cavity lengths. The figure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km fiber. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance bl f l f h f f h OA h d k Figure 8. Measured SNR to compare the impact of transmission ﬁber on the performance of an RSOA-FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non-zero dispersion shifted ﬁber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km ﬁber cavity lengths. The ﬁgure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km ﬁber. Figure 8. Measured SNR to compare the impact of transmission fiber on the performance of an RSOA- FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non- zero dispersion shifted fiber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km fiber cavity lengths. The figure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km fiber. Figure 8. Measured SNR to compare the impact of transmission ﬁber on the performance of an RSOA-FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non-zero dispersion shifted ﬁber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km ﬁber cavity lengths. The ﬁgure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km ﬁber. Figure 8. Measured SNR to compare the impact of transmission fiber on the performance of an RSOA- FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non- Figure 8. Measured SNR to compare the impact of transmission ﬁber on the performance of an Figure 8. Measured SNR to compare the impact of transmission fiber on the performance of an RSOA- FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non- zero dispersion shifted fiber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km fiber cavity lengths. The figure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km fiber. Figure 8. 4.2. RSOA-FCL Performance 4.2. RSOA-FCL Performance Measured SNR to compare the impact of transmission ﬁber on the performance of an RSOA-FCL: The solid and dotted lines show the measured SNR for BtB (back-to-back) and after 25 km non-zero dispersion shifted ﬁber (NZDSF) transmission, respectively. In both cases, the results are for 15 m, 115 m and 1 km ﬁber cavity lengths. The ﬁgure shows that the SNR bandwidth narrows by up to 50% after transmission over 25 km ﬁber. y p , cavities for back-to-back (BtB) and 25 km non-zero dispersion shifted fiber (NZDSF) transmission. RSOA-FCL Cavity Length Tx Peak SNR (dB) Usable SNR Bandwidth (GHz) 15 m BtB 19.45 6.7 25 km NZDSF 19.01 3.2 115 m BtB 19.5 6.4 25 km NZDSF 19.2 3.2 1 km BtB 17.7 4.5 25 km NZDSF 17.1 2.8 4.3. Performance Comparison of the Three Transmitter Schemes Here, we compared the SNR performance of the ASE spectrum-sliced with the self-seeded RSOA-FCL and the externally seeded RSOA sources. For this experiment, the ASE spectrum-sliced source was equipped with the 2 nm filter similar to that of the RSOA-FCL For the RSOA-FCL we Table 2. Summary of results for the SNR performance of the RSOA-FCL with 15 m, 115 m and 1 km cavities for back-to-back (BtB) and 25 km non-zero dispersion shifted ﬁber (NZDSF) transmission. RSOA-FCL Cavity Length Tx Peak SNR (dB) Usable SNR Bandwidth (GHz) 15 m BtB 19.45 6.7 25 km NZDSF 19.01 3.2 115 m BtB 19.5 6.4 25 km NZDSF 19.2 3.2 1 km BtB 17.7 4.5 25 km NZDSF 17.1 2.8 4.3. Performance Comparison of the Three Transmitter Schemes y p , cavities for back-to-back (BtB) and 25 km non-zero dispersion shifted fiber (NZDSF) transmission. RSOA-FCL Cavity Length Tx Peak SNR (dB) Usable SNR Bandwidth (GHz) Table 2. Summary of results for the SNR performance of the RSOA-FCL with 15 m, 115 m and 1 km cavities for back-to-back (BtB) and 25 km non-zero dispersion shifted ﬁber (NZDSF) transmission. RSOA-FCL and the externally seeded RSOA sources. For th source was equipped with the 2 nm filter, similar to that of 4.3. Performance Comparison of the Three Transmitter Schemes took the 15 m and 1 km cavities, which offered the best and the worst performance, respectively. Both RSOA-FCLs were equipped with a 2 nm filter. The RSOA input and output power for the externally seeded scheme was −3 dBm, and ~6 dBm, respectively. The bias current for all experiments was set to 100 mA. Figure 9a,b show the measured SNR values for BtB and transmission over 25 km NZDSF, Here, we compared the SNR performance of the ASE spectrum-sliced with the self-seeded RSOA-FCL and the externally seeded RSOA sources. For this experiment, the ASE spectrum-sliced source was equipped with the 2 nm ﬁlter, similar to that of the RSOA-FCL. For the RSOA-FCL, we took the 15 m and 1 km cavities, which offered the best and the worst performance, respectively. Both 11 of 17 ers a 6.7 on with Appl. Sci. 2017, 7, 999 and 2.8 GHz for th GHz bandwidth RSOA-FCLs were equipped with a 2 nm ﬁlter. The RSOA input and output power for the externally seeded scheme was −3 dBm, and ~6 dBm, respectively. The bias current for all experiments was set to 100 mA. Figure 9a,b show the measured SNR values for BtB and transmission over 25 km NZDSF, respectively. It can be seen that the externally seeded RSOA and the self-seeded RSOA roughly offer a 10 dB and a 5 dB higher SNR over the ASE spectrum-sliced source across the whole spectral band. The self-seeded RSOA-FCL with the shorter cavity outperforms the longer cavity FCL by about 1–2 dB across the spectral band. And while the externally seeded RSOA exhibited the highest SNR bandwidth of ~9.6 GHz for BtB, the 15 m and 1 km long RSOA-FCLs offer useable bandwidths of 6.7 and 4.5 GHz and the ASE spectrum-sliced scheme features a usable bandwidth of only 2.5 GHz. p y p ( y ) dispersion of the transmission fiber. The narrower usable bandwidth recorded after transmission will influence the maximum achievable bit rate. We attribute the higher performance observed for the RSOA-FCL when compared to the ASE spectrum-sliced source to three main factors. First, the RSOA-FCL has a significantly narrower spectrum, which significantly reduces the impact of chromatic dispersion [17,50]. Second, the RSOA-FCL has a larger output power. For example, at 100 mA, the RSOA-FCL has an output power of −3.4 dBm while the ASE spectrum sliced scheme emits −11.8 dBm. RSOA-FCL and the externally seeded RSOA sources. For th source was equipped with the 2 nm filter, similar to that of 4.3. Performance Comparison of the Three Transmitter Schemes Third, the saturated RSOA in the cavity nonlinearly suppress the amplitude noise in the lasing cavity, which lowers the RIN. These three contribute to the improve SNR of the RSOA-FCL compared to the ASE spectrum sliced source [17]. Figure 9. Comparison of the measured SNR for the three colorless transmitter schemes: RSOA ASE spectrum-sliced (SS), self-seeded RSOA-FCL and externally seeded (ES) RSOA. (a) Shows the BtB (back-to-back) SNR measurement. The externally seeded RSOA (green line) outperforms the RSOA- FCL (red and brown) and the ASE spectrum-sliced scheme (blue) by ~10 and ~5 dB SNR across the whole frequency range, respectively; (b) Shows the SNR after 25 km NZDSF transmission. The SNR bandwidth is reduced due to dispersion of the transmission fiber. In general, the RSOA-FCL offers performance midway between an ASE spectrum sliced and an externally seeded RSOA. Figure 9. Comparison of the measured SNR for the three colorless transmitter schemes: RSOA ASE spectrum-sliced (SS), self-seeded RSOA-FCL and externally seeded (ES) RSOA. (a) Shows the BtB (back-to-back) SNR measurement. The externally seeded RSOA (green line) outperforms the RSOA-FCL (red and brown) and the ASE spectrum-sliced scheme (blue) by ~10 and ~5 dB SNR across the whole frequency range, respectively; (b) Shows the SNR after 25 km NZDSF transmission. The SNR bandwidth is reduced due to dispersion of the transmission ﬁber. In general, the RSOA-FCL offers performance midway between an ASE spectrum sliced and an externally seeded RSOA. Figure 9. Comparison of the measured SNR for the three colorless transmitter schemes: RSOA ASE Figure 9. Comparison of the measured SNR for the three colorless transmitter schemes: RSOA spectrum-sliced (SS), self-seeded RSOA-FCL and externally seeded (ES) RSOA. (a) Shows the BtB (back-to-back) SNR measurement. The externally seeded RSOA (green line) outperforms the RSOA- FCL (red and brown) and the ASE spectrum-sliced scheme (blue) by ~10 and ~5 dB SNR across the whole frequency range, respectively; (b) Shows the SNR after 25 km NZDSF transmission. The SNR bandwidth is reduced due to dispersion of the transmission fiber. In general, the RSOA-FCL offers performance midway between an ASE spectrum sliced and an externally seeded RSOA. ASE spectrum-sliced (SS), self-seeded RSOA-FCL and externally seeded (ES) RSOA. (a) Shows the BtB (back-to-back) SNR measurement. RSOA-FCL and the externally seeded RSOA sources. For th source was equipped with the 2 nm filter, similar to that of 4.3. Performance Comparison of the Three Transmitter Schemes The source of the SNR degradation with transmission is the interplay of the chirp (induced by the direct modulation of the RSOA) and the dispersion of the transmission ﬁber. The narrower usable bandwidth recorded after transmission will inﬂuence the maximum achievable bit rate. 5.1. Maximizing the Capacity of the RSOA-FCL In our experiments, we performed bit- and power-loading based on Chows algorithm [38]. Chow’s algorithm can briefly be described as follows. It takes in the SNR characterization of a transmitter at the subcarrier frequencies of the DMT signal. It also uses the SNR requirement of all advanced modulation formats (see Figure 3) to achieve a predetermined BER. Then, it disregards We attribute the higher performance observed for the RSOA-FCL when compared to the ASE spectrum-sliced source to three main factors. First, the RSOA-FCL has a signiﬁcantly narrower spectrum, which signiﬁcantly reduces the impact of chromatic dispersion [17,50]. Second, the RSOA-FCL has a larger output power. For example, at 100 mA, the RSOA-FCL has an output power of −3.4 dBm while the ASE spectrum sliced scheme emits −11.8 dBm. Third, the saturated RSOA in the cavity nonlinearly suppress the amplitude noise in the lasing cavity, which lowers the RIN. These three contribute to the improve SNR of the RSOA-FCL compared to the ASE spectrum sliced source [17]. RSOA-FCL and the externally seeded RSOA sources. For th source was equipped with the 2 nm filter, similar to that of 4.3. Performance Comparison of the Three Transmitter Schemes The externally seeded RSOA (green line) outperforms the RSOA-FCL (red and brown) and the ASE spectrum-sliced scheme (blue) by ~10 and ~5 dB SNR across the whole frequency range, respectively; (b) Shows the SNR after 25 km NZDSF transmission. The SNR bandwidth is reduced due to dispersion of the transmission ﬁber. In general, the RSOA-FCL offers performance midway between an ASE spectrum sliced and an externally seeded RSOA. 5. Bit- and Power-Loaded DMT for Highest Bit Rates In this section, bit- and power-loaded DMT transmission is used to maximize the data throughput for any of the three bandwidth limited sources. We proceed in two steps. We again first study the impact of the cavity length on the RSOA-FCL and the achievable data throughput in more detail. Only when we know the optimum operation conditions of the RSOA-FCL, we investigate and compare the maximum capacity of all three schemes. When transmitting the signals over 25 km NZDSF ﬁber, the usable SNR bandwidth reduced to ~1.6 GHz for the ASE spectrum-sliced source. The RSOA-FCL declines to a usable bandwidths of 3 and 2.8 GHz for the 15 m and 1 km long cavities. And the externally seeded RSOA still offers a 6.7 GHz bandwidth with sufﬁcient SNR for transmission. The source of the SNR degradation with transmission is the interplay of the chirp (induced by the direct modulation of the RSOA) and the dispersion of the transmission ﬁber. The narrower usable bandwidth recorded after transmission will inﬂuence the maximum achievable bit rate. 5. Bit- and Power-Loaded DMT for Highest Bit Rates In this section, bit- and power-loaded DMT transmission is used to maximize the data throughput for any of the three bandwidth limited sources. We proceed in two steps. We again first study the impact of the cavity length on the RSOA-FCL and the achievable data throughput in more detail. Only when we know the optimum operation conditions of the RSOA-FCL, we investigate and compare the maximum capacity of all three schemes. When transmitting the signals over 25 km NZDSF ﬁber, the usable SNR bandwidth reduced to ~1.6 GHz for the ASE spectrum-sliced source. The RSOA-FCL declines to a usable bandwidths of 3 and 2.8 GHz for the 15 m and 1 km long cavities. And the externally seeded RSOA still offers a 6.7 GHz bandwidth with sufﬁcient SNR for transmission. 5.1. Maximizing the Capacity of the RSOA-FCL In our experiments, we performed bit- and power-loading based on Chows algorithm [38]. Chow’s algorithm can brieﬂy be described as follows. It takes in the SNR characterization of a transmitter at the subcarrier frequencies of the DMT signal. It also uses the SNR requirement of all advanced modulation formats (see Figure 3) to achieve a predetermined BER. Then, it disregards subcarriers with the lowest SNR that cannot support transmission at the predetermined BER for even the simplest format. The power of such unusable subcarriers is then transferred to usable subcarriers. For subcarrier with non-integer bit per symbol, the bit number is rounded off to the nearest integer, which requires the power to be re-adapted accordingly. This power adaptation step (power-loading) helps to achieve similar BER for all subcarriers. Appl. Sci. 2017, 7, 999 12 of 17 subcarriers with the lowest SNR that cannot support transmission at the predetermined BER for even the simplest format. The power of such unusable subcarriers is then transferred to usable subcarriers. For subcarrier with non-integer bit per symbol, the bit number is rounded off to the nearest integer, First, we performed bit- and power-loaded DMT transmission for the RSOA-FCL with 15 m, 115 m and 1 km cavities. The signal was optimized for a BtB experiment and for transmission over 25 km NZDSF. Figure 10a–c show the SNR, and the bit- as well as the power-loading patterns of the DMT and the measured per-subcarrier BER for 15 m, 115 m and 1 km long RSOA-FCLs, respectively. The 15 and 115 m cavities can support up to 5 bit/symbol (32QAM) for low frequency subcarriers. In case of the 1 km RSOA-FCL, the highest order modulation format is 16QAM, with 4 bits/symbol. The power-loading pattern is arranged to guarantee a minimum SNR difference for all subcarriers with the same modulation format, so that a similar BER performance can be obtained. The solid lines in the BER plots indicate the overall BER, i.e., the total BER across all subcarriers. It can be seen that the overall BER is below 9.29 × 10−3 in all instances, which is the limit for an FEC (forward error correction) with 12.5% overhead [54]. which requires the power to be re-adapted accordingly. This power adaptation step (power-loading) helps to achieve similar BER for all subcarriers. 5.1. Maximizing the Capacity of the RSOA-FCL First, we performed bit- and power-loaded DMT transmission for the RSOA-FCL with 15 m, 115 m and 1 km cavities. The signal was optimized for a BtB experiment and for transmission over 25 km NZDSF. Figure 10a–c show the SNR, and the bit- as well as the power-loading patterns of the DMT and the measured per-subcarrier BER for 15 m, 115 m and 1 km long RSOA-FCLs, respectively. The 15 and 115 m cavities can support up to 5 bit/symbol (32QAM) for low frequency subcarriers. In case of the 1 km RSOA-FCL, the highest order modulation format is 16QAM, with 4 bits/symbol. The power-loading pattern is arranged to guarantee a minimum SNR difference for all subcarriers with the same modulation format, so that a similar BER performance can be obtained. The solid lines in the BER plots indicate the overall BER, i.e., the total BER across all subcarriers. It can be seen that the overall BER is below 9.29 × 10−3 in all instances, which is the limit for an FEC (forward error correction) with 12.5% overhead [54]. Figure 10. Measured SNR, bit- and power-loading pattern, and bit error ratio (BER) versus sub-carrier frequencies and the achieved total line rate for different RSOA-FCL cavity lengths: Plots (a–c) show the results for 15, 115 m and 1 km long cavities, respectively. The red and orange colors indicate BtB (back-to-back) and 25 km NZDSF transmission measurements. In all cases, the overall BER (solid lines in the BER plots) are kept below the FEC limit of 9.29 × 10−3 [53]; (d) Plots the measured total line rate for 15, 115 m and 1 km long cavities, both for BtB and after 25 km transmission. In a BtB measurement, the RSOA-FCL offers up to 20 Gbit/s line rates. The line-rate is still 10 Gbit/s when transmitting signals over 25 km. Figure 10d shows the maximum line rates obtained with RSOA-FCL cavity lengths of 15 m, 115 m and 1 km length for a BtB and a transmission over 25 km NZDSF Maximum capacities are obtained Figure 10. Measured SNR, bit- and power-loading pattern, and bit error ratio (BER) versus sub-carrier frequencies and the achieved total line rate for different RSOA-FCL cavity lengths: Plots (a–c) show the results for 15, 115 m and 1 km long cavities, respectively. The red and orange colors indicate BtB (back-to-back) and 25 km NZDSF transmission measurements. 5. Bit- and Power-Loaded DMT for Highest Bit Rates In this section, bit- and power-loaded DMT transmission is used to maximize the data throughput for any of the three bandwidth limited sources. We proceed in two steps. We again ﬁrst study the impact of the cavity length on the RSOA-FCL and the achievable data throughput in more detail. Only when we know the optimum operation conditions of the RSOA-FCL, we investigate and compare the maximum capacity of all three schemes. Appl. Sci. 2017, 7, 999 12 of 17 12 of 17 5.2. Comparison of Maximizing Capacity 5.2. Comparison of Maximizing Capacity Second, we compared the maximum capacity achievable with bit- and power-loaded ASE spectrum-sliced, two types of self-seeded FCL RSOAs and externally seeded RSOAs. Second, we compared the maximum capacity achievable with bit- and power-loaded ASE spectrum-sliced, two types of self-seeded FCL RSOAs and externally seeded RSOAs. Second, we compared the maximum capacity achievable with bit- and power-loaded ASE spectrum-sliced, two types of self-seeded FCL RSOAs and externally seeded RSOAs. Second, we compared the maximum capacity achievable with bit- and power-loaded ASE spectrum-sliced, two types of self-seeded FCL RSOAs and externally seeded RSOAs. Figure 11a,b show BtB and 25 km DMT transmission results, respectively. For low subcarrier frequencies, the ASE spectrum-sliced source supported up to 3 bit/symbol (8QAM), the RSOA-FCL up to 5 bit/symbol (32QAM), and the externally seeded RSOA up to 6 bit/symbol (64QAM). The power-loading pattern shown in Figure 11a,b led to relatively similar SNRs for subcarriers with the same modulation format. As a result, the BER for different subcarriers was relatively constant over the modulation frequency. The solid lines show the overall BER. For all schemes, the overall BERs were kept below 7.5 × 10−3 (10% overhead FEC) [54]. Figure 11a,b show BtB and 25 km DMT transmission results, respectively. For low subcarrier frequencies, the ASE spectrum-sliced source supported up to 3 bit/symbol (8QAM), the RSOA-FCL up to 5 bit/symbol (32QAM), and the externally seeded RSOA up to 6 bit/symbol (64QAM). The power-loading pattern shown in Figure 11a,b led to relatively similar SNRs for subcarriers with the same modulation format. As a result, the BER for different subcarriers was relatively constant over the modulation frequency. The solid lines show the overall BER. For all schemes, the overall BERs were kept below 7.5 × 10−3 (10% overhead FEC) [54]. Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. 5.1. Maximizing the Capacity of the RSOA-FCL In all cases, the overall BER (solid lines in the BER plots) are kept below the FEC limit of 9.29 × 10−3 [53]; (d) Plots the measured total line rate for 15, 115 m and 1 km long cavities, both for BtB and after 25 km transmission. In a BtB measurement, the RSOA-FCL offers up to 20 Gbit/s line rates. The line-rate is still 10 Gbit/s when transmitting signals over 25 km. Figure 10. Measured SNR, bit- and power-loading pattern, and bit error ratio (BER) versus sub-carrier frequencies and the achieved total line rate for different RSOA-FCL cavity lengths: Plots (a–c) show the results for 15, 115 m and 1 km long cavities, respectively. The red and orange colors indicate BtB (back-to-back) and 25 km NZDSF transmission measurements. In all cases, the overall BER (solid lines in the BER plots) are kept below the FEC limit of 9.29 × 10−3 [53]; (d) Plots the measured total line rate for 15, 115 m and 1 km long cavities, both for BtB and after 25 km transmission. In a BtB measurement, the RSOA-FCL offers up to 20 Gbit/s line rates. The line-rate is still 10 Gbit/s when transmitting signals over 25 km. Figure 10d shows the maximum line rates obtained with RSOA-FCL cavity lengths of 15 m, 115 Figure 10. Measured SNR, bit- and power-loading pattern, and bit error ratio (BER) versus sub-carrier frequencies and the achieved total line rate for different RSOA-FCL cavity lengths: Plots (a–c) show the results for 15, 115 m and 1 km long cavities, respectively. The red and orange colors indicate BtB (back-to-back) and 25 km NZDSF transmission measurements. In all cases, the overall BER (solid lines in the BER plots) are kept below the FEC limit of 9.29 × 10−3 [53]; (d) Plots the measured total line rate for 15, 115 m and 1 km long cavities, both for BtB and after 25 km transmission. In a BtB measurement, the RSOA-FCL offers up to 20 Gbit/s line rates. The line-rate is still 10 Gbit/s when transmitting signals over 25 km. Appl. Sci. 2017, 7, 999 13 of 17 13 of 17 Figure 10d shows the maximum line rates obtained with RSOA-FCL cavity lengths of 15 m, 115 m and 1 km length for a BtB and a transmission over 25 km NZDSF. Maximum capacities are obtained for short FCL cavities of 15 and 115 m length. 5.1. Maximizing the Capacity of the RSOA-FCL The capacities are between 20 and 10 Gbit/s depending on the transmission distance. For longer cavity lengths the capacity dropped to 12.4 Gbit/s due to a reduced SNR and a limited bandwidth for the BtB experiment and 7 Gbit/s could at best be transmitted over 25 km. Appl. Sci. 2017, 7, 999 13 of 17 for short FCL cavities of 15 and 115 m length. The capacities are between 20 and 10 Gbit/s depending on the transmission distance. For longer cavity lengths the capacity dropped to 12.4 Gbit/s due to a reduced SNR and a limited bandwidth for the BtB experiment and 7 Gbit/s could at best be transmitted over 25 km. 5.2. Comparison of Maximizing Capacity 5.2. Comparison of Maximizing Capacity All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Figure 11. 6. Conclusions We show how transmission capacities of low-cost low-quality sources can be optimized for maximum capacities by means of bit- and power-loaded DMT transmission. Record high transmission data rates for an ASE spectrum-sliced, self-seeded RSOA-FCL and externally seeded RSOA of 6.25, 20 and 30.7 Gbit/s in BtB and 4.2, 12.4 and 24.5 Gbit/s over 25 km NZDSF transmission are reported. To the best of our knowledge, these are the highest capacities reported for such inexpensive and colorless sources. Our experiments show that even lowest cost sources such as spectral sliced ASE sources or RSOA-FCLs can offer quite some capacity if properly operated, for example, for O-band upstream transmission. More precisely, ASE sources can provide capacities in the range of 6 to 4 Gbit/s depending on the distance. Fabry-Perot sources such as the RSOA-FCL may offer capacities in the order of 20 to 10 Gbit/s for distances up to 25 km. Finally, more costly externally seeded colorless sources may provide capacities in the range of 30 to 25 Gbit/s if operated with an optimized bit- and power loaded DMT transmission format without offset ﬁltering. Acknowledgments: This work was supported by the EU FP7 projects ERMES (grant agreement no. 288542) and Fox-C (grant agreement No. 318415). Sterlite Technologies Ltd. is acknowledged for providing the ﬁber. Author Contributions: Simon Arega Gebrewold conceived the concept, designed and performed the experiments, carried out the signal processing and analysis and wrote the paper. Romain Bonjour contributed in the concept development, experiment and manuscript writing. Romain Brenot designed and fabricated the RSOAs. David Hillerkuss organized the experiment and wrote the paper. Juerg Leuthold conceived and developed the concept, designed the experiment, wrote the paper and supervised the work. Conﬂicts of Interest: The authors declare no conﬂict of interest. 5.2. Comparison of Maximizing Capacity 5.2. Comparison of Maximizing Capacity This can be explained by the relatively wider linewidth of the two sources, thus incurring higher dispersion penalty. However, O-band RSOAs [53], can be used to lower dispersion penalty and avoid the need for NZDSF. O-band has already been used for upstream transmission in legacy PONs to relax the requirement on inexpensive poor sources [1,55]. It is interesting to note that the chirp-dispersion induced usable bandwidth reduction results in considerable capacity reduction, especially in the case of the ASE spectrum-sliced and RSOA-FCL transmitters. This can be explained by the relatively wider linewidth of the two sources, thus incurring higher dispersion penalty. However, O-band RSOAs [53], can be used to lower dispersion penalty and avoid the need for NZDSF. O-band has already been used for upstream transmission in legacy PONs to relax the requirement on inexpensive poor sources [1,55]. Table 3. Summary of achieved line rate and the respective bit error ratio (BER) for the three transmitter schemes. The results are for a BtB (back-to-back) and after 25 km NZDSF transmission. ASE: ampliﬁed spontaneous emitters. Table 3. Summary of achieved line rate and the respective bit error ratio (BER) for the three transmitter schemes. The results are for a BtB (back-to-back) and after 25 km NZDSF transmission. ASE: ampliﬁed spontaneous emitters. ASE Spectrum-Sliced RSOA-FCL (15 m) RSOA-FCL (1 km) Externally Seeded RSOA Line Rate (Gbit/s) BER Line Rate (Gbit/s) BER Line Rate (Gbit/s) BER Line Rate (Gbit/s) BER BtB 6.25 5.8 × 10−3 20.1 5.8 × 10−3 12.4 5.8 × 10−3 30.7 5.7 × 10−3 25 km NZDSF 4.2 2.3 × 10−3 10.1 4 × 10−3 7 5.8 × 10−3 24.5 7.1 × 10−3 5.2. Comparison of Maximizing Capacity 5.2. Comparison of Maximizing Capacity Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Figure 11. Measured SNR, bit- and power-loading pattern and per-subcarrier BER for the three transmitter schemes: RSOA ASE spectrum-sliced (SS) (blue), self-seeded RSOA-FCL (red and brown) and externally seeded (ES) RSOA (green). (a) and (b) show results for back to back case and 25 km transmission. The ASE spectrum sliced scheme can support modulation formats with up to 3 bits/symbol while RSOA-FCL and externally seeded RSOA can encode up to 6 bits/symbol for low subcarrier frequencies. The solid line in the bottom plot indicates the overall BER that can be achieved with either of the three transmission schemes. All bit-error ratios are within the FEC (forward error correctio) limit of 7.5 × 10−3. Table 3 summarizes the achieved maximum line rates for the three transmitter schemes and the corresponding overall BER. In a BtB experiment the ASE spectrum-sliced, the RSOA-FCL and the externally seeded sources were delivering capacities up to 6.25, 20.1 and 30.7 Gbit/s, respectively. For a link with 25 km the sources could still deliver maximum capacities of 4.2, 10.1 and 24.5 Gbit/s. It is interesting to note that the chirp-dispersion induced usable bandwidth reduction results in Table 3 summarizes the achieved maximum line rates for the three transmitter schemes and the corresponding overall BER. In a BtB experiment the ASE spectrum-sliced, the RSOA-FCL and the externally seeded sources were delivering capacities up to 6.25, 20.1 and 30.7 Gbit/s, respectively. For a link with 25 km the sources could still deliver maximum capacities of 4.2, 10.1 and 24.5 Gbit/s. 14 of 17 Appl. 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https://openalex.org/W4396572262	OpenAlex	Open Science	CC-By	2,022	ESCUTAÇÃO: ENCONTROS ENTRE CRIANÇAS, ASSISTÊNCIA SOCIAL E PANDEMIA	Letícia Maísa Eichherr	Portuguese	Spoken	7,898	17,057	ESCUTAÇÃO: ENCONTROS ENTRE CRIANÇAS, ASSISTÊNCIA SOCIAL E PANDEMIA “ESCUCHA-ACCIÓN”: ENCUENTROS ENTRE NIÑOS, ASISTENCIA SOCIAL Y PANDEMIA “LISTENING-ACTION”: ENCOUNTERS BETWEEN CHILDREN, SOCIAL ASSISTANCE AND PANDEMIC ARTIGO Letícia Maísa Eichherr e Lílian Rodrigues da Cruz http://dx.doi.org/10.1590/1807-0310/2022v34263894 Universidade Federal do Rio Grande do Sul, Porto alegre/RS, Brasil Resumo: Neste artigo objetiva-se discutir questões relacionadas aos encontros das crianças com a assistência social a partir de um recorte da pesquisa de mestrado realizada em um Serviço de Convivência e Fortalecimento de Vínculos. Apostamos nos traçados singulares da metodologia cartográfica para o acompanhamento dos acontecimentos cotidianos do serviço e para o exercício epistêmico de descolonização do pensamento. Pretendemos, assim, contextualizar os discursos vinculados às crianças consideradas público prioritário do serviço e objetos da política pública e refletir sobre a pandemia da Covid-19 que escancara as desigualdades sociais. Entendemos que o olhar crítico desde uma perspectiva parcial potencializa a desconstrução de discursos totalizantes e universais sobre as infâncias e propomos a escutação como aposta ética-estética-política de uma escuta menor em tempos e sistemas maiores, que podem brutalizar o cotidiano de trabalho nas políticas públicas. p p Palavras-chave: Política pública; Criança; Psicologia social; Assistência social; Covid-19. Resumen : Este artículo tiene como objetivo discutir cuestiones relacionadas con los encuentros de los niños con la asistencia social, a partir de un fragmento de una investigación de maestría realizada en un Servicio de Convivencia y Fortalecimiento de Vínculos. Apostamos por las huellas únicas de la metodología cartográfica para el seguimiento de los acontecimientos cotidianos del servicio y para el ejercicio epistémico de descolonización del pensamiento. Pretendemos, por tanto, contextualizar los discursos vinculados a los niños considerados público prioritario del servicio y objeto de la política pública y reflexionar sobre la pandemia de la Covid-19, que abre las desigualdades sociales. Entendemos que la mirada crítica desde una perspectiva parcial potencia la deconstrucción de discursos totalizadores y universales sobre la infancia y proponemos la “escucha-acción” como una apuesta ético-estética-política de una escucha más pequeña en tiempos y sistemas más grandes que puede embrutecer el trabajo cotidiano en las políticas públicas. Palabras clave: Política pública; Niño; Psicología social; Asistencia social; Covid-19. Abstract: This article aims to discuss issues related to children’s encounters with social assistance, based on an excerpt from a master's research carried out in the Service for Coexistence and Strengthening of Bonds. We bet on the unique traces of the cartographic methodology for the monitoring of the daily events of the service and for the epistemic exercise of decolonization of thought. Introdução Neste artigo, apresentamos um recorte de uma pesquisa que se compôs nas tessituras dos encontros das crianças com as políticas públicas de assistência social em tempos de pandemia da Covid-19, mais especificamente em um Serviço de Convivência e Fortalecimento de Víncu los (SCFV) do município de Porto Alegre/RS. Contextualizamos alguns enquadres discursivos que podem direcionar a estigmas e este reótipos colados às crianças consideradas público prioritário do SCFV, refletimos sobre o tempo da pandemia que escancara as desigualdades sociais e a necropolítica da sociedade brasileira e propomos criançamentos na assistência social inventando a palavra escutação para falar de uma escuta menor em tempos maiores que brutalizam o cotidiano de trabalho nas políticas públicas. Criançamento é uma palavra inventada pelo poeta Manoel de Barros (2010, p. 339) que queria “chegar ao criançamento das palavras”, fazer uma revolução da linguagem e brincar com o sentido acostumado das palavras. Neste artigo, fazer criançamento é uma tentativa de desconstrução dos discursos que enquadram as infâncias em modelos universais para abrir-se ao encontro com a po tência do que elas dizem, fazem, veem acerca do mundo, tal qual o menino que, “diante de uma casa em demolição, observa: ‘Olha, pai! Estão fazendo um terreno!’” (Rosa, 1967/2009, pp. 30-31). ESCUTAÇÃO ESCUTAÇÃO ISSN 1807-0310 ISSN 1807-0310 ESCUTAÇÃO: ENCONTROS ENTRE CRIANÇAS, ASSISTÊNCIA SOCIAL E PANDEMIA “ESCUCHA-ACCIÓN”: ENCUENTROS ENTRE NIÑOS, ASISTENCIA SOCIAL Y PANDEMIA “LISTENING-ACTION”: ENCOUNTERS BETWEEN CHILDREN, SOCIAL ASSISTANCE AND PANDEMIC We intend, therefore, to contextualize the speeches linked to children considered a priority public of the service and objects of public policy and to reflect on the Covid-19 pandemic that opens up social inequalities. We understand that the critical look from a partial perspective enhances the deconstruction of totalizing and universal discourses about childhood. Besides, we propose listening-action as an ethical-aesthetic-political bet of a smaller listening in larger times and systems, which can brutalize the daily work in public policies. Keywords: Public Policy; Child; Social Psychology; Social Assistance; Covid-19. Este artigo está licenciado sob uma Licença Creative Commons. Com essa licença você pode compartilhar, adaptar, para qualquer fim, desde que atribua a autoria da obra, forneça um link para a licença, e indicar se foram feitas alterações. ESCUTAÇÃO ESCUTAÇÃO Esses critérios específicos podem gerar estereótipos e práticas menoristas, porém os mo dos de implementar e executar as políticas públicas podem configurar-se como resistência à própria história na qual elas se constituem, marcadas por contradições e ideias normatizado ras. Nesse sentido, Lílian Cruz, Betina Hillesheim e Neuza Guareschi (2005) discorrem sobre a história das práticas psicológicas nas políticas públicas e como contribuem para legitimar as “crianças como objeto psico-médico-biológico, passíveis de serem medidas, testadas, ordenadas e denominadas normais e anormais” (p. 44) e, com isso, produzem “subjetividades desqualifi cadas – famílias incompetentes e negligentes – ... e uma infância tida como ideal, desejável, normal” (p. 46). De forma semelhante, Cecília Coimbra e Maria do Nascimento (2008) ressal tam que as práticas e teorias cientificistas, racistas e eugênicas atuam na produção de subje tividades “que têm caracterizado a população infanto-juvenil subalternizada como perigosa, violenta, criminosa e, muitas vezes, percebida como não humana” (p. 1). Os SCFV estão inseridos no mesmo território de abrangência dos CRAS que se localizam em áreas de maior vulnerabilidade social de um município (MDS, 2014; Política Nacional de Assistência Social - PNAS, 2004). Estão nas vilas, nas comunidades periféricas, nas favelas, numa atuação com a infância considerada pobre e que é “alvo maciço dos saberes/poderes modernos, a qual é compreendida como um problema político e econômico, que exige esforços no sentido de seu ordenamento e controle” (Hillesheim & Cruz, 2008, p. 195). Com as atividades do SCFV, consideradas de caráter preventivo, há uma tentativa de gestão dos riscos sociais e, pela probabi lidade de prever acontecimentos, busca-se modificar o que é indesejado com ações mitigadoras, buscando a antecipação do futuro numa intervenção no presente (Hillesheim & Cruz, 2008). Simone Hüning, Rosângela Cabral e Maria Ribeiro (2018) destacam que os territórios, principalmente da assistência social, se constituem pelas desigualdades no investimento e no governo das cidades e, ligado a isso, há discursos totalizantes de desqualificação e criminaliza ção direcionados aos moradores, que incidem na execução das políticas públicas, perpassadas pela racialização, por preconceitos e pela idealização de modos de vida. Notas sobre o Serviço de Convivência e Fortalecimento de Vínculos O SCFV é um serviço da Política Nacional de Assistência Social (PNAS) ofertado de forma complementar ao trabalho social com famílias realizado nos Centros de Referência da Assistência Social (CRAS) e nos Centros de Referência Especializado da Assistência Social (CREAS) com “vistas a prevenir a ocorrência de situações de risco social e fortalecer os vínculos familiares e co munitários” (Ministério do Desenvolvimento Social - MDS, 2016, p. 13). É organizado em gru pos, dividido por ciclos de vida, observando as necessidades, especificidades e diversidades das/ os participantes, tendo um máximo de 30 usuários sob mediação de uma educadora social, com encontros regulares e atividades planejadas (MDS, 2016, 2017a, 2017b). Segundo as orientações técnicas do extinto Ministério do Desenvolvimento Social e Combate à Fome (MDS, 2017a, p. 8), “o SCFV possui um caráter preventivo e proativo, pautado na defesa e afirmação de direitos e no desenvolvimento de capacidades e potencialidades dos usuários, com vistas ao alcance de alternativas emancipatórias para o enfrentamento das vulnerabilidades sociais”. A pesquisa que embasa este artigo foi realizada em um SCFV do município de Porto Ale gre/RS, com crianças e adolescentes de 6 a 14 anos, tendo aprovação do Comitê de Ética em Pesquisa do Instituto de Psicologia da Universidade Federal do Rio Grande do Sul. Em Porto Alegre/RS, os SCFV atendem aproximadamente dez mil crianças e adolescentes de 6 a 15 anos1. Com o início da pandemia da Covid-19, as atividades presenciais do SCFV foram suspensas durante alguns meses, sendo retomadas no final de 2020 e organizadas para garantir o distanciamen to social: horários reduzidos, poucas crianças simultaneamente e agendamento prévio. Anteriormen te, eram em dias úteis, turnos diários, inversos ao da escola, de até quatro horas (MDS, 2014). O público prioritário do SCFV são crianças e adolescentes que vivenciam situações de trabalho infantil, violências, negligências, fora da escola, defasagem escolar, acolhimento ins titucional, egressos ou em cumprimento de medida socioeducativa, com medidas de proteção, situação de rua, entre outras (MDS, 2016; 2017a; 2017b). 2 ISSN 1807-0310 2 ISSN 1807-0310 ISSN 1807-0310 ESCUTAÇÃO Dessa forma, o SCFV, assim como outros serviços da assistência social, precisa de insis tência “em um fazer sensível às diferenças entre territorialidades e as experiências tantas vezes invisibilizadas”, tal qual relatado por Hüning et al. (2018, p. 66). Um exercício que perpassa a ideia de que os modos de execução das políticas públicas podem ser resistência às durezas cotidianas e às macropolíticas que objetificam as crianças. As trabalhadoras atuantes nesses es paços podem fazer o jogo de reprodução de modelos e métodos que não permitem criar saídas para os processos de singularização e de produção de desejos, mas também podem trabalhar numa micropolítica que constrói novos modos de subjetividade (Guattari & Rolnik, 1985), potencializando intervenções em que a voz das crianças seja escutada, suas problematizações e perspectivas de olhar consideradas e seus modos de vida não criminalizados. Ademais, a insistência e a resistência também se relacionam com as (im)possibilidades de efeti var a política pública de assistência social diante de tempos de desmontes, uma vez que ao olharmos os documentos técnicos utilizados nessa escrita, datados de um período em que havia no governo federal o Ministério do Desenvolvimento Social (de 2004 a 2018), responsável por políticas sociais e por uma ampla produção de conhecimento e orientações às trabalhadoras da PNAS, entendemos que a redução deste ministério a uma secretaria do Ministério da Cidadania no atual (des)governo Bolsonaro, desde janeiro de 2019, é simbólica. A conhecida PEC da Morte (Emenda Constitucional nº 95 de 2016) e a Portaria nº 2362 de 23 de dezembro de 2019, que reduz o repasse de verbas aos municípios, são exemplos dos cortes orçamentários, do desfinanciamento e da inconstância de re cursos e investimentos que acontecem desde o golpe de 2016 e que impactam diretamente na oferta dos serviços previstos na PNAS, consequentemente no SCFV, nos programas sociais como o Bolsa Família e o Benefício de Prestação Continuada, recoloca o Brasil no Mapa da Fome da Organização das Nações Unidas e “traz de volta práticas conservadoras da assistência social, como o primeiro damismo e práticas moralizantes e higienizadoras de controle da população pobre” (Delgado, To lentino, Barbosa, Machado, & Nunes, 2022, pp. 43-44). ESCUTAÇÃO Em consonância à proteção social e à prevenção de riscos sociais, é indicado que a atuação do SCFV nos territórios seja de forma a identificar suas potencialidades e de valorizá-los enquanto lu gar de pertença dos sujeitos (MDS, 2017b), sendo fundamental compreender que os territórios são produzidos nas tramas e tessituras das interações e “das relações cotidianas, seus modos de vida, sua configuração topográfica e morfológica, suas condições de urbanização e infraestrutura urbana, suas condições de moradia que compõem o cenário e a história ali vivida” (MDS, 2017b, p. 52). Entretanto, um dos desafios do SCFV em realizar ações cuidadosas, acolhedoras, sobretudo não coloniais ou violentas, é perpassado por discursos de ódio internalizados na sociedade, rela cionados à produção de crianças e jovens perigosos em determinados territórios e que se mate rializam em falas propagadas por cidadãos de bem, como um dos que encontramos durante a pes quisa e que diz o quão importante é ter um espaço como o SCFV para as crianças não ficarem na rua e não virarem bandidos. Discursos que produzem subjetividades no contemporâneo e que são direcionados às crianças do SCFV que têm em seus corpos a inscrição da intersecção dos marca dores sociais da diferença de raça, gênero, classe e território e que, tantas vezes, são enquadrados como alvos da necropolítica, objetos de uma política pública e público prioritário de um serviço. Cabe pontuar que os mesmos documentos que apontam para critérios e enquadramentos trazem pistas sobre a importância de um espaço sensível e do olhar às singularidades, desta cando que “promover bons encontros, que fortaleçam a potência de agir pode impulsionar a ação para enfrentar situações conflituosas, alterar condições de subordinação, estabelecer diá logos, desejar e atuar por um mundo mais digno e mais justo” (MDS, 2017b, p. 28). ISSN 1807-0310 3 ISSN 1807-0310 ESCUTAÇÃO “Como o que se passa entre é o mais interessante, resta ao cartógrafo estar suficientemente poroso a estas microssensibilidades que se instauram nas zonas fronteiriças” (Costa, 2014, p. 67). Zonas essas que são acontecimentos da própria vida e que convocam às apostas em políticas do criançar e criançamentos, para uma desconstrução dos discursos totalizantes sobre as infâncias. Leni Dornelles (2010, p. 17) ressalta que as crianças têm sido “fabricadas como sujeitos que escolhem, decidem, optam e de alguma maneira muito cedo precisam assumir os efeitos de suas decisões” que as enquadram em infâncias emergentes na atualidade, como as daguerra, dareli giosidade, daciberinfância, e outras, no contexto que pesquisamos: doSASE3, davila, ou com ou tros adjetivos que podem colar e produzir mais enquadres, como criança-abusada, criança-pobre, criança-vulnerável, criança-em-risco. Mesmo que esses adjetivos não sejam uma escolha, vive mos um tempo que individualiza a questão social e culpabiliza os sujeitos, inclusive as crianças. Sabemos que o encontro das crianças com o SCFV acontece por motivos potencialmente tristes, como descrito anteriormente. Considerando que o SCFV é complementar ao trabalho social realizado com famílias, as crianças que ali chegam costumam ser encaminhadas por ou tros serviços, como CRAS, CREAS, serviço de abordagem social de rua, acolhimento institu cional e a partir de discussões de casos com a rede intersetorial que busca um espaço protetivo às crianças que vivenciam situações de vulnerabilidades sociais e violação de direitos. Vivências que não são estáticas e não devem ser definidoras de estereótipos das crianças doSASE, aquelas também nomeadas em risco, potencialmente perigosas, que precisam ser tu teladas, que precisam de intervenção e condução para que sobrevivam à infância, uma etapa da vida a ser superada, e tornem-se adultos trabalhadores. Entretanto, para que não se reproduzam esses discursos no cotidiano do SCFV, no planeja mento das atividades e nas discussões entre profissionais e serviços, é necessário um constante exercício de perguntar-se e de olhar os atravessamentos do “sistema-mundo capitalista, patriar cal, eurocêntrico, cristão, moderno e colonialista” (Grosfoguel, 2016, p. 32) na nossa constituição enquanto sujeitos, que se expressam na forma como trabalhamos, escutamos e intervimos, reco nhecendo que há uma produção social de subalternidades que afetam diretamente as crianças que se encontram no SCFV, assim como estruturam infâncias desejáveis e universais. Cartografar e criançar A cartografia, enquanto escolha metodológica desta pesquisa, se dá como uma afirmação da potência da heterogeneidade da vida e acontece “no plano da experiência, lá onde conhecer e fazer se tornam inseparáveis, impedindo qualquer pretensão à neutralidade” (Passos & Bar ros, 2015, p. 30). É considerada uma pesquisa intervenção, com uma inevitável implicação dos envolvidos, que borra as fronteiras “de quem conhece e do que é conhecido, de quem analisa e do que é analisado” (Passos & Barros, 2015, p. 25). O traçado singular do percurso cartográfico desta pesquisa se deu a partir da inserção da primeira autora no SCFV, durante quatro meses (dezembro de 2020 a março de 2021), acompa nhando os encontros entre crianças, trabalhadoras e acontecimentos do cotidiano do serviço, com registros em diário de campo, fotografias, desenhos e escritas das crianças. Festa de Natal, almoço de Ano Novo, caminhar pelo território, pintar pneu para fazer um vaso de flor, cuidar do pátio, desenhar com aquarela, tecer filtro dos sonhos e mandalas, jogar UNO, Fla-Flu2 e xadrez, fazer roda de conversa na sala e no pátio, levar os trabalhos finais na escola, torcer para passar de ano, comer melancia gelada, filmar as piruetas no campo de futebol e o gol de bicicleta, fazer silêncio no refeitório, escrever pela primeira vez em um com putador, lavar as mãos e passar álcool em gel, higienizar a mesa, escutar uma história, contar outra história, sorrir com os olhos, ter esperança que a pandemia acabe. ISSN 1807-0310 ESCUTAÇÃO Obirin Odara (2021, s/p) pontua o quão colonial é associar às crianças a ideia de seres faltantes, que precisam crescer, superar a infância e viver a adultidade como auge do tempo cronológico da vida. A questão é que a criança não será algo quando crescer porque não devemos projetá-la como alguém que ainda deve conquistar seu lugar no mundo, fazendo dela um sujeito que vive à espera do depois. O que elas têm a contribuir com nossa comunidade agora, a partir desse lugar mágico que é a infância? Talvez a pergunta que precise ser feita é para os adultos: o que vocês foram e não são mais? A criança já é. Reflexões que convocam à descolonização do nosso olhar para “a concepção iluminista da infância e de sua educação como preparação para o futuro, para o adulto que será, o que desapropria a experiência humana das crianças” (Faria, Barreiro, Macedo, & Santos, 2015, p. 14). Às crianças, como se concordássemos com a ausência de sua fala e de um lugar próprio e possível no mundo, desejamos que mais rapidamente cresçam e sejam o futuro da nação. Anete Abramowicz e Tatiane Rodrigues (2014, p. 464) dizem que “há um preceito hegemônico que fala de infância e da potência da criança sob os auspícios da economia, como investimento social e capital do futuro”. Hillesheim e Cruz (2008, p. 197) apontam que “ao relacionar os fatores (de risco) do presente, busca-se fabricar o futuro: a infância (hoje) de risco deve ser transformada no adulto produtivo (de amanhã)”. As crianças que encontramos no SCFV, nomeadas por muitos de vulneráveis e em risco, fa lam de outras coisas, de muitas coisas. Falam de seus modos de ser criança neste tempo indigesto e sombrio que parece ser o presente, muito mais do que o futuro. Assim, optamos “pelo presente possível, pela utopia do aqui agora, ao invés da ‘velha’ utopia futurista de que o ‘amanhã será melhor’ e [que por isso] devemos nos esforçar para crescer; preferimos nos manter ‘crianças’” (Noguera & Barreto, 2018, p. 641). Afinal, nas suas diferenças e singularidades, as crianças nos movimentam a pensar na potência das diferentes infâncias, que não cabem num conceito único, mesmo que pareçam ter que caber num espaço-tempo delimitado, aquele do zero aos 12 anos. ESCUTAÇÃO Apostamos no tempo de criançar, na potência da criança em afetar e ser afetada, sua dis ponibilidade de construir mundos, disposição para invenção, para imprevistos e para surpre ender-se (Vicentin, 2020). Um verbo que abre passagem aos movimentos das crianças e das infâncias no contemporâneo. Que deixam entrar em cena as perguntas que as próprias crianças nos fazem acerca das perspectivas de futuro que a adultidade tem negligenciado. ESCUTAÇÃO Betina Hillesheim e Neuza Guareschi (2007) pontuam que, pela busca da universalidade necessária no projeto moderno de ciência, que homogeneíza sujeitos, minimizando as diferen ças e maximizando as semelhanças, a Psicologia acabou legitimando um modelo de infância ideal que exclui possibilidades outras de ser criança através de “uma produção discursiva que constitui, a partir de suas descrições, uma infância desejada, normal, ajustada e, da mesma for ma, indesejada, patológica, desajustada” (Hillesheim & Guareschi, 2007, p. 88). Sabemos, sobretudo, que a produção de conhecimentos em nosso sistema-mundo, não so mente sobre as infâncias, é constituída a partir de determinadas escolhas epistemológicas em de trimento de outras, sustentadas pelo colonialismo e pelo racismo que se “constituíram num apa rato global de destruição de corpos, mentes e espíritos”, como aponta Sueli Carneiro (2005, pp. 101-102). Assim, olhar as infâncias de uma perspectiva parcial e situada requer um deslocamento epistêmico e uma abertura para questionamentos acerca da naturalidade com que as olhamos e a universalidade das verdades ditas e produzidas sobre elas. Donna Haraway (1995/2009, p. 25) destaca que “a visão é sempre uma questão do poder de ver - e talvez da violência implícita em nossas práticas de visualização. Com o sangue de quem foram feitos os meus olhos?”. Refletimos, com isso, sobre uma pergunta que é direcionada às crianças: o que você quer ser quando crescer? ISSN 1807-0310 ESCUTAÇÃO ESCUTAÇÃO O seu morro é uma comunidade quilombola que a cada chuva convive com o risco de desmo ronamento. Um morro invisibilizado pelo poder público, sem acessibilidade, sem transporte público. Já Menino E disse que não tinha sonhos. Era difícil sonhar lá do alto do morro. Da sua janela vê o movimento da polícia, os meninos do tráfico. Sua mãe lhe diz que precisa andar bem arrumado. Talvez uma das suas preocupações esteja relacionada aos dados do Anuário Brasileiro de Segurança Pública de 2021 sobre mortes violentas intencionais de crianças e ado lescentes no ano de 2020, que apontam que “a partir dos 10 anos, mas ainda mais fortemente a partir dos 15 anos, o perfil das vítimas se torna predominantemente de pessoas negras do sexo masculino” (Reinach, 2021, p. 232), com aumento da letalidade por intervenção policial. É a materialidade da interseccionalidade dos marcadores sociais da diferença de raça, gênero, classe e território, como apontado por Carla Akotirene (2019), que escancara que os meninos, negros, pobres e da periferia são os principais alvos das diferentes violências que assolam o Brasil, dentre elas a violência policial. Achille Mbembe (2016) desenvolve o conceito de necropolítica que nos ajuda a entender as dinâmicas de uma racionalidade soberana e colonial que produz políticas de morte, ditando quais são os corpos que podem viver e quais devem morrer, criando, assim, mundos de morte e inimigos ficcionais que podem ser exterminados a qualquer custo e a qualquer momento. Ao falar sobre a ocupação colonial contemporânea, Mbembe aponta para a centralidade da raça e do racismo que estruturam a submissão de determinados corpos ao poder soberano que define “quem importa e quem não importa, quem é ‘descartável’ e quem não é” (Mbembe, 2016, p. 135). Renato Noguera (2020, 2021) nomeia de necroinfância o dispositivo da necropolítica que tem como objeto as crianças negras, destituindo-lhes a vida e produzindo crianças matáveis, invisíveis, violáveis e que nascem com um alvo no peito. O autor nos convoca à urgência de um pacto político-social pela vida das crianças negras, tendo em vista as desigualdades sociais que as afetam, seja no acesso à escola ou à saúde, seja pela alta vitimização em assassinatos e outras violências. Conceição Evaristo (2017, p. 84) ecoa a necroinfância em poesia denúncia quando diz: “pedimos que as balas perdidas percam o nosso rumo e não façam do corpo nosso, os nossos filhos, o alvo”. ISSN 1807-0310 Tempos de pandemia e da necropolítica Boaventura Santos (2020), ao refletir sobre a pandemia do coronavírus, pontua a exis tência de zonas de invisibilidade estruturadas pelo capitalismo, o colonialismo e o patriarcado, que produzem sombras e ausência de comoção, e que essas “zonas de invisibilidade poderão multiplicar-se em muitas outras regiões do mundo, e talvez mesmo aqui, bem perto de cada um de nós. Talvez baste abrir a janela” (Santos, 2020, p. 9). É improrrogável abrir a janela, as cortinas e os vidros que podem criar dificuldades à visão. Menino E e Diego4, crianças que encontrei no SCFV, falavam de sonhos. Diego sonhava com uma casa que não fosse no morro, mesmo que de lá veja todas as estrelas no horizonte. ISSN 1807-0310 ISSN 1807-0310 6 ESCUTAÇÃO ESCUTAÇÃO dos familiares, da rede de apoio, das escolas; muitas não tiveram a garantia da possibilidade do isolamento social e dos cuidados sanitários preconizados; a perda econômica nas famílias, que fez aumentar a pobreza, o trabalho infantil e a insegurança alimentar; a dificuldade no acesso ao ensino e a consequente evasão escolar; a morte de familiares, em que muitas crianças ficaram órfãs; rupturas nas interações sociais; aumento das violências domésticas e intrafami liares, com diminuição das denúncias. dos familiares, da rede de apoio, das escolas; muitas não tiveram a garantia da possibilidade do isolamento social e dos cuidados sanitários preconizados; a perda econômica nas famílias, que fez aumentar a pobreza, o trabalho infantil e a insegurança alimentar; a dificuldade no acesso ao ensino e a consequente evasão escolar; a morte de familiares, em que muitas crianças ficaram órfãs; rupturas nas interações sociais; aumento das violências domésticas e intrafami liares, com diminuição das denúncias. Uma pandemia que escancara as desigualdades sociais e os abismos já existentes no Bra sil. O sucateamento das políticas públicas, a má gestão do combate à pandemia e a precarização de determinadas vidas são retratos da necropolítica e do racismo estrutural que é considerado “uma forma sistemática de discriminação que tem a raça como fundamento, e que se manifesta por meio de práticas conscientes ou inconscientes que culminam em desvantagens ou privilégios para indivíduos, a depender do grupo racial ao qual pertençam” (Almeida, 2018, p. 25). Diego entendia bem sobre isso. Contou que dava para perceber que os árbitros de futebol dão mais cartão vermelho para os jogadores negros5. Mesmo que não nomeie de racismo estrutural, seu corpo sabe o que é. Também sabe do isolamento social que sua comunidade quilombola vive no alto do morro, mesmo antes da pandemia. Então, quando falamos que as crianças são o futuro da nação, a quais crianças direcionamos o olhar, as expectativas de futuro e o direito de sonhar? Barros (2010, p. 172) conta que um “homem tinha sido escolhido, desde criança, para ser ninguém e nem nunca. De forma que quando se pensou em fazer alguma coisa por ele, viu-se que o caso era irremediável e escuro”. ISSN 1807-0310 ESCUTAÇÃO Maya Angelou (1978/2018, s/p) fala, também, de um racismo cotidia no vivenciado pelas crianças negras: “Na escola nova, um pesadelo, meninos puxam meu cabelo (meninas imbatíveis de cabelos crespos incríveis), eles não me assustam nada”. Este pode ser um jeito para Diego responder ao tema que buscou na escola: escolha três monstros e escreva por que tem medo deles. As opções xerocadas numa folha: Drácula, Frankens tein, fantasma, Coringa, lobisomem, e alguns outros mais. Ele dizia que não sabia quais escolher, talvez porque já haja tantos monstros reais que esses da lista realmente não assustam nada. Ao longo do ano de 2020, foi difícil imaginar as crianças sem o encontro com as profes soras, educadoras, colegas, amigos da escola e do SCFV. Distantes das poéticas públicas (Ca ppellari, 2019), das apostas micropolíticas que se fazem nos encontros com as políticas públi cas. Passamos, então, a escutar histórias de um tempo pandêmico quando as crianças retornam fisicamente aos serviços e transbordam os acontecimentos da vida em palavras, pois eles já não se sustentam apenas na memória. Menino E conheceu o pai na pandemia. Já se falavam pelo facebook. Contou que seu pai é branco e que sua mãe é parda e que se parece mais com o pai. Para Diego, foi difícil fazer os trabalhos da escola durante a pandemia, pois não tinha internet. Menino E também não. Diego queria saber quantas pessoas tem no mundo hoje, pois muitas morreram pelo coronavírus. Isabel Brandão (2021) e Márcia Buss-Simão e Juliana Lessa (2020) relatam alguns impac tos da pandemia na vida das crianças, como o isolamento social que provocou o afastamento ISSN 1807-0310 ISSN 1807-0310 ESCUTAÇÃO (p. 56); valorização, como “estratégia que considera as questões e problemas do outro como pro cedentes e legítimos” (p. 57); reconhecimento dos limites e das possibilidades das situações vivi das por si e pelos outros; experiência de reconhecer e nomear suas emoções, buscando aprender sobre os sentimentos, afetações, disparadores de sensações negativas e intensas. (p. 56); valorização, como “estratégia que considera as questões e problemas do outro como pro cedentes e legítimos” (p. 57); reconhecimento dos limites e das possibilidades das situações vivi das por si e pelos outros; experiência de reconhecer e nomear suas emoções, buscando aprender sobre os sentimentos, afetações, disparadores de sensações negativas e intensas. Escutação: Por uma escuta menor “Tudo que eu tinha ouvido sobre eles era como eram pobres, assim havia se tornado im possível pra mim vê-los como alguma coisa além de pobres. Sua pobreza era minha história única sobre eles” (Adichie, 2009, transcrição de vídeo). Chimamanda Adichie (2009), ao falar do perigo de uma história única e contar da sua experiência com a família de Fide, parece di zer um tanto das experiências vivenciadas na assistência social, em que é pressuposto escutar histórias e que, muitas delas, são contadas por outros que não os personagens principais. Judi ciário, conselho tutelar, escola, unidade de saúde. Quando se trata de histórias de crianças, pelo viés da proteção integral e da garantia de direitos, há grandes chances da escuta ser perpassada por palavras fatídicas: risco, violência, negligência. A escutação não é apenas uma palavra inventada, é uma tentativa ética-estética-política de delirar as reflexões sobre as possibilidades dos encontros com outros, outras, outres, em um co tidiano de escuta na assistência social. A escutação não é isenta, não é neutra, é atravessada pelo silêncio, pelos afetos, pelo lugar social ocupado por quem escuta. É construída numa relação de corpos, de saberes, de poderes e pode ser patriarcal, colonial, capitalista, eurocêntrica, racista e sexista (Grosfoguel, 2016). Fazer escutação diz da necessidade de perceber, acompanhar e interrogar os processos subjetivos e sociais que transversalizam a escuta e o corpo de pesqui sadora, escutadora, psicóloga, educadora. É necessário um olhar atento para si, para estranhar-se, reconhecer privilégios e para o que se reproduz no lugar que se ocupa. Há sempre que se considerar ilusões de ótica, que podem ser causadas por lentes da branquitude, do racismo, do adultocentrismo, da cishetero normatividade. Podemos atentar-se a isto com algumas perguntas cotidianas: “Como ver? De onde ver? Quais os limites da visão? Ver para quê? Ver com quem? Quem deve ter mais do que um ponto de vista? Nos olhos de quem se joga areia? Quem usa viseiras? Quem interpreta o campo visual?” (Haraway, 1995/2009, p. 28). É necessário um olhar atento para si, para estranhar-se, reconhecer privilégios e para o que se reproduz no lugar que se ocupa. Há sempre que se considerar ilusões de ótica, que podem ser causadas por lentes da branquitude, do racismo, do adultocentrismo, da cishetero normatividade. Podemos atentar-se a isto com algumas perguntas cotidianas: “Como ver? De onde ver? Quais os limites da visão? Ver para quê? Ver com quem? ESCUTAÇÃO Lembremos que nós, pessoas brancas, estruturadas na branquitude, somos historicamente quem têm o poder de escolher e não escolher histórias, tendo em vista que o racismo estrutural privilegia pessoas brancas (Kilomba, 2008/2019), que não se veem como racializadas e sim como sujeitos universais e superiores (Schucman, 2012), onde “o silêncio, a omissão, a distorção do lugar do branco na situação das desigualdades ra ciais no Brasil têm um forte componente narcísico, de autopreservação” (Bento, 2014, p. 30). Enquanto pesquisadoras e profissionais operadoras das políticas públicas, há que se des colonizar o olhar e a escuta, desaprender o racismo, fazer algo desde o nosso lugar social. Ha raway (1995/2009, p. 30) argumenta “a favor de políticas e epistemologias de alocação, posicio namento e situação”, pontuando que a perspectiva parcial e localizada requer responsabilidade “tanto pelas suas promessas quanto por seus monstros destrutivos” (p. 21). Santos (2020, p. 15) destaca que “qualquer quarentena é sempre discriminatória, mais difícil para uns grupos sociais do que para outros” e designa o Sul como um espaço-tempo po lítico, social e cultural e uma “metáfora do sofrimento humano injusto causado pela exploração capitalista, pela discriminação racial e pela discriminação sexual”. Apesar do autor não nomear as crianças como um grupo específico que compõe esse Sul da pandemia, compreendemos que elas estão em todos os grupos descritos por ele, afetadas pelas suas condições enquanto mu lheres, trabalhadores precarizados, moradores das periferias pobres das cidades, a população de rua, os refugiados e imigrantes, as pessoas com deficiência, os idosos. Perguntamo-nos, assim, quais são os resquícios de uma pandemia na vida das crianças do SCFV. Quais vivências são possíveis de virar palavras? Quais permanecem inscritas em seus corpos, mas que precisam um pouco mais de encontros e escutas para transbordar? Há pistas para o SCFV nos documentos orientadores (MDS, 2017b), escritos antes da pan demia e que sustentam que o SCFV é lugar de acolhimento e de afeto e de produzir jeitos possí veis, micropolíticos, nas brechas, nas frestas, nas rachaduras das concretudes cotidianas: escuta, em que há “interesse na história e apreço pelo trajeto vivido pelo sujeito que narra, busca dos motivos e não das justificativas, busca do entendimento e não do julgamento sobre as situações” ISSN 1807-0310 ISSN 1807-0310 ESCUTAÇÃO que criam zonas de invisibilidade e silenciamento e como abrir brechas, rachaduras nas dure zas históricas e estruturais. São exercícios cotidianos de descolonização do pensamento. que criam zonas de invisibilidade e silenciamento e como abrir brechas, rachaduras nas dure zas históricas e estruturais. São exercícios cotidianos de descolonização do pensamento. A escuta menor está interessada nas luzes, histórias, vozes menores, na zona onde navegam as forças, as intensidades, os voos, as viagens silenciosas, isto é, os escapes de saúde. Uma escuta que não procura formatar uma identidade, mas conhecer as forças intrusas que habitam em nós. (Serfaty, 2016, p. 13) Escutação é escuta+ação+olhar que escapam e resistem aos usos acostumados e acomo dados dos ouvidos, da boca, dos olhos, das mãos que escrevem relatórios, dos pés que cami nham nos territórios rumo às visitas domiciliares. Sobretudo, não é preciso ver, ouvir, falar, andar, pois se faz com o corpo que se tem, transita entre territórios físicos e subjetivos, dos jeitos possíveis e no tempo necessário, com todos os sentidos, não só os conhecidos. Escutação: Por uma escuta menor Quem deve ter mais do que um ponto de vista? Nos olhos de quem se joga areia? Quem usa viseiras? Quem interpreta o campo visual?” (Haraway, 1995/2009, p. 28). A escutação, uma escuta-ação-cuidado vai se tecendo, sobretudo, de um modo menor, aquele minoritário, do qual Gilles Deleuze e Félix Guattari (1975/2017) nos falam. Escutação é escuta menor, opera em devir minoritário, um acontecimento do encontro com outro, outra, outres. É o que se passa entre-dois, em que há dupla captura - dos que escutam, dos escutados; pode ser silencioso, quase imperceptível. Não há modelo a seguir, nem jeitos certos ou erra dos. É devir. “Os devires são geografia, são orientações, direções, entradas e saídas” (Deleuze & Parnet, 1980/1998, p. 02). Sendo devir, sempre muda, tanto quanto muda a intercessora, interlocutora, escutadora, não as imita, mas inventa. Tecemos a escuta menor junto da literatura menor de Deleuze e Guattari (1975/2017), que dizem que “uma literatura menor não é a de uma língua menor, mas antes a que uma mi noria faz em uma língua maior” (Deleuze & Guattari, 1975/2017, p. 30). Busca-se, desse modo, pistas para um exercício minoritário em uma escuta maior, que é perpassado por reconhecer o que são os maiores que nos atravessam, quais são os saberes e poderes engendrados na escuta ISSN 1807-0310 9 ISSN 1807-0310 Criançamentos finais As crianças foram mostrando, ao longo da pesquisa e na sutileza dos encontros no SCFV, que o falatório teórico dos adultos pode tornar-se colonizador e opressor, buscando enquadres às vivências e modos de estar no mundo, numa justificativa de criar possibilidades da existên cia de um futuro a partir de uma intervenção individual com discurso motivacional que facil mente cai na lógica da meritocracia. Ao questionar os instituídos, as rotinas estabelecidas, as próprias perguntas feitas, transfor mando-as em outras perguntas, as normas que as encapsulam num tempo cronológico para ser criança, as crianças nos mostram que viver a infância no morro, na vila, na rua, na sinaleira, na pandemia, na escola, no SCFV, no acolhimento institucional, não as fazem menos crianças ou com menos infância. Elas falam não só em palavras, mas em gestos, olhares, desenhos, das singulari dades de suas histórias e convocam a um olhar descolonizador das infâncias, a contrapelo daquela majoritária, desejada e propagada socialmente, com aberturas às epistemologias ameríndias, afri canas, latino-americanas, feministas, como uma escolha ética e política de resistência. É pela escutação que confirmamos o que Deleuze e Guattari (1975/2017, p. 42) diziam: “não há tão grande, nem revolucionário, quanto o menor”. Ao propormos a escutação como uma escuta menor, entendemos que há uma dimensão individual do fazer profissional ético cotidiano, mesmo diante da precarização do trabalho e do desfinanciamento da política de assistência social, como aposta na possibilidade de perguntar-se, reconhecer perspectivas de visão e privilégios, e identificar discursos totalizantes direcionados às crianças que se encontram não apenas nos ser viços da assistência social, mas também em tantos outros lugares, como na escola, na mídia, nas redes sociais. Discursos totalizantes que seguem sustentando modelos universais, eurocêntricos, brancos, ideais de viver e reproduzindo perguntas e modos de lidar com as crianças na perspec tiva de almejar um futuro e um vir a ser dentro desses mesmos padrões. Afinal, perguntamos “o que você quer ser quando crescer?” esperando determinadas respostas dentro de um viés econô mico, de trabalho, de formação universitária, de sucesso profissional. Escutação faz delirar verbos, embaralhar equações, inventar sonhos, “sonhar o contrário: saber criar um devir-menor” (Deleuze & Guattari, 1975/2017, p. 43). ISSN 1807-0310 10 ESCUTAÇÃO ESCUTAÇÃO Notas 1 Informação em: http://www2.portoalegre.rs.gov.br/fasc/default.php?p_secao=1488 Informação em: http://www2.portoalegre.rs.gov.br/fasc/default.php?p_secao=1488 2 Fla-Flu é a forma como o jogo de futebol de mesa é nomeado no Rio Grande do Sul. Em outros lugares do Brasil também é chamado de pebolim, totó ou pacau. 2 Fla-Flu é a forma como o jogo de futebol de mesa é nomeado no Rio Grande do Sul. Em outros lugares do Brasil também é chamado de pebolim, totó ou pacau. 3 SASE é a sigla de Serviço de Apoio Sócio Educativo, como era nomeado o SCFV antes da publicação da Tipificação Nacional dos Serviços Socioassistenciais de 2009. 3 SASE é a sigla de Serviço de Apoio Sócio Educativo, como era nomeado o SCFV antes da publicação da Tipificação Nacional dos Serviços Socioassistenciais de 2009. 3 SASE é a sigla de Serviço de Apoio Sócio Educativo, como era nomeado o SCFV antes da publicação da Tipificação Nacional dos Serviços Socioassistenciais de 2009. 4 Nomes fictícios. 5 Conversa após o jogo de futebol entre Paris Saint Germain e Basaksehir, onde o jogador camaronês Webó recebeu cartão vermelho. Notícia https://ge.globo.com/futebol/futebol-in ternacional/liga-dos-campeoes/noticia/uefa-suspende-cartao-vermelho-recebido-por-webo -na-partida-entre-psg-e-basaksehir.ghtml 5 Conversa após o jogo de futebol entre Paris Saint Germain e Basaksehir, onde o jogador camaronês Webó recebeu cartão vermelho. Notícia https://ge.globo.com/futebol/futebol-in ternacional/liga-dos-campeoes/noticia/uefa-suspende-cartao-vermelho-recebido-por-webo -na-partida-entre-psg-e-basaksehir.ghtml ISSN 1807-0310 11 ISSN 1807-0310 11 ESCUTAÇÃO Referências Abramowicz, Anete & Rodrigues, Tatiane C. (2014). Descolonizando as pesquisas com crianças e três obstáculos. Educação & Sociedade [online], 35(127), 461-474. https://doi. org/10.1590/S0101-73302014000200007 Adichie, Chimamanda (2009). O perigo de uma única história [vídeo]. Conferência de TE DGlobal. https://www.ted.com/talks/chimamanda_ngozi_adichie_the_danger_of_a_single_ story?language=pt Akotirene, Carla (2019). Interseccionalidade. Pólen. Almeida, Silvio (2018). Racismo estrutural. Letramento. Angelou, Maya (1978/2018). A vida não me assusta. (A. Paiva, trad.). DarkSide Books. (Ori ginal publicado em 1978) Barros, Manoel de (2010). Poesia completa. Leya. Bento, Maria A. (2014). Branqueamento e branquitude no Brasil. In I. Carone & M. Bento, (Orgs.), Psicologia Social do Racismo: Estudos sobre branquitude e branqueamento no Brasil (pp. 25-58). Vozes. Brandão, Isabel (2021). Infância em tempos de pandemia. HOLOS, 3, 1-17. https://doi. org/10.15628/holos.2021.11951 Buss-Simão, Márcia & Lessa, Juliana S. (2020). Um olhar para o(s) corpo(s) das crianças em tempos de pandemia. Zero-a-Seis, 22(nspe.), 1420-1445. https://doi.org/10.5007/1980-4512. 2020v22nespp1420 Cappellari, Amanda (2019). Poéticas insurgentes: o desligamento institucional por maiori dade [Dissertação de Mestrado, Psicologia Social e Institucional, Universidade Federal do Rio Grande do Sul, Porto Alegre/RS]. Carneiro, Aparecida Sueli (2005). A Construção do Outro como Não-Ser como fundamento do Ser [Tese de doutorado, Programa de Pós-graduação em Educação, Universidade de São Paulo, São Paulo/SP]. Coimbra, Cecília & Nascimento, Maria L. (2008). A produção de crianças e jovens perigo sos: a quem interessa? CEDECA – São Martinho. http://www.infancia-juventude.uerj.br/pdf/ livia/aproducao.pdf i Costa, Luciano B. (2014). Cartografia: uma outra forma de pesquisar. Revista Digital do LAV, 7(2) 65-67. https://doi.org/10.5902/1983734815111 Cruz, Lílian; Hillesheim, Betina, & Guareschi, Neuza (2005). Infância e políticas públicas: um olhar sobre as práticas psi. Psicologia & Sociedade [online], 17(3), 42-49. https://doi.org/10.1590/S0102-71822005000300006 Delgado, Diana, Tolentino, Erika S., Barbosa, Mara C. F., Machado, Ricardo W. G., & Nunes, Nilza R. A. (2022). (Des)financiamento e (des)proteção social: o abate da ‘prima pobre’ da Seguridade Social. O Social em Questão, 25(52), 35-60. https://doi.org/10.17771/ PUCRio.OSQ.56391 Deleuze, Gilles & Guattari, Félix (1975/2017). Kafka: por uma literatura menor (C. V. Sil va, trad.). Autêntica. (Original publicado em 1975) Deleuze, Gilles & Parnet, Claire (1980/1998). Diálogos. (E. A. Ribeiro, trad.). Escuta. e, Gilles & Guattari, Félix (1975/2017). Kafka: por uma literatura menor (C. V. Sil ). Autêntica. (Original publicado em 1975) Deleuze, Gilles & Parnet, Claire (1980/1998). Diálogos. (E. A. Ribeiro, trad.). Escut Deleuze, Gilles & Parnet, Claire (1980/1998). Diálogos. (E. A. Ribeiro, trad.). Escuta. Dornelles, Leni V. (2010). Sobre o devir-criança ou discursos sobre as infâncias. Referências In Anais do V Colóquio Internacional de Filosofia da Educação. Universidade Estadual do Rio de Janeiro. https://www ufrgs br/gein/wp-content/uploads/2016/10/sobre-o-devir-crian%C3%A7a-ou-dis Dornelles, Leni V. (2010). Sobre o devir-criança ou discursos sobre as infâncias. In A V Colóquio Internacional de Filosofia da Educação. Universidade Estadual do Rio de Janeiro. https://www.ufrgs.br/gein/wp-content/uploads/2016/10/sobre-o-devir-crian%C3%A7a-ou-dis cursos-sobre-as-infancias.pdf Evaristo, Conceição (2017). Poemas da recordação e outros movimentos. Malê. ISSN 1807-0310 12 ESCUTAÇÃO Faria, Ana L. G., Barreiro, Alex, Macedo, Eliana E., & Santos, Solange E. (2015). Invita ções Pós-coloniais. In (Orgs.), Infâncias e pós-colonialismo: pesquisas em busca de pedago gias descolonizadoras (pp. 11-24). Leitura Crítica; Associação de Leitura do Brasil.i (pp ) uattari, Félix & Rolnik, Suely (1985). Micropolítica: cartografias do desejo. Vozes. Grosfoguel, Ramón (2016). A estrutura do conhecimento nas universidades ocidentalizadas: racismo/sexismo epistêmico e os quatro genocídios/epistemicídios do longo século XVI. So ciedade e Estado [online], 31(1), 25-49. https://doi.org/10.1590/S0102-69922016000100003 Haraway, Donna (1995/2009). Saberes localizados: a questão da ciência para o feminismo e o privilégio da perspectiva parcial. Cadernos Pagu, 5, 7-41. https://periodicos.sbu.unicamp.br/ ojs/index.php/cadpagu/article/view/1773 Hillesheim, Betina & Guareschi, Neuza (2007). De que infância nos fala a psicologia do de senvolvimento?: Algumas reflexões. Psicologia da Educação, 25, 75-92. http://pepsic.bvsalud. org/scielo.php?script=sci_arttext&pid=S1414-69752007000200005&lng=pt&tlng=pt Hillesheim, Betina & Cruz, Lílian (2008). Risco, vulnerabilidade e infância: algumas apro ximações. Psicologia & Sociedade [online], 20(2), 192-199. https://doi.org/10.1590/S0102- 71822008000200006 Hüning, Simone, Cabral, Rosângela J., & Ribeiro, Maria A. T. (2018). Nas Margens: Psi cologia, Política de Assistência Social e Territorialidades. Revista Polis e Psique, 8(3), 52-69. https://dx.doi.org/10.22456/2238-152X.86108 Kilomba, Grada (2008/2019). Memórias da Plantação: episódios de racismo cotidiano (J. Oliveira, trad.). Cobogó. Mbembe, Achille (2016). Necropolítica. Arte & Ensaio. Revista do PPGAV/EBA/UFRJ, 32, 123-151. https://revistas.ufrj.br/index.php/ae/article/view/8993/7169 i Ministério do Desenvolvimento Social e Combate à Fome – MDS (2014). Tipificação Na i l d S i S i i t i i Autor Ministério do Desenvolvimento Social e Combate à Fome – MDS (2014). Tipificação Na cional de Serviços Socioassistenciais. Autor. ional de Serviços Socioassistenciais. Autor. Ministério do Desenvolvimento Social e Combate à Fome – MDS. (2016). Caderno de Orientações: Serviço de Proteção e Atendimento Integral à Família e Serviço de Convivência e Fortalecimento de Vínculos. Autor. Orientações: Serviço de Proteção e Atendimento Integral à Família e Serviço de Convivência e Fortalecimento de Vínculos. Autor. ( ) g quentes: Serviço de Convivência e Fortalecimento de Vínculos (SCFV). Autor. Ministério do Desenvolvimento Social – MDS (2017b). Concepção de convivência e forta lecimento de vínculos. Autor. Noguera, Renato & Barreto, Marcos (2018). LETÍCIA EICHHERR LETÍCIA EICHHERR https://orcid.org/0000-0002-6840-9342 Mestra em Psicologia Social e Institucional pela Universidade Federal do Rio Grande do Sul (PPGPSI/UFRGS). E-mail: leticiae.psico@gmail.com Mestra em Psicologia Social e Institucional pela Universidade Federal do Rio Grande do Sul (PPGPSI/UFRGS). E-mail: leticiae.psico@gmail.com Referências Infancialização, Ubuntu e Teko Porã: elementos gerais para educação e ética afroperspectivistas. Childhood & Philosophy, 14(31), 625-644. https://doi.org/10.12957/childphilo.2018.36200 Noguera, Renato (2020, 09 de julho). Necroinfância: por que as crianças negras são assassina das? Portal Lunetas. https://lunetas.com.br/necroinfancia-criancas-negras-assassinadas Noguera, Renato (2021, 08 de janeiro). Crianças negras desaparecidas: até quando vamos aceitar? Portal Lunetas. https://lunetas.com.br/criancas-negras-desaparecidas-ate-quando-vamos-aceitar das? Portal Lunetas. https://lunetas.com.br/necroinfancia-criancas-negras-assassinadas Noguera, Renato (2021, 08 de janeiro). Crianças negras desaparecidas: até quando vamos aceitar? Portal Lunetas. https://lunetas.com.br/criancas-negras-desaparecidas-ate-quando-vamos-aceitar Odara, Obirin. @naomecolonize (2021, 12 de outubro). O vício colonial da falta/incomple tude se expressa também na pergunta feita às crianças: “o que você quer ser quando crescer?”. Instagram. https://www.instagram.com/p/CGQ6OaFpVo_/ i Odara, Obirin. @naomecolonize (2021, 12 de outubro). O vício colonial da falta/incomple tude se expressa também na pergunta feita às crianças: “o que você quer ser quando crescer?”. Instagram. https://www.instagram.com/p/CGQ6OaFpVo_/ i Passos, Eduardo & Barros, Regina B. (2015). A cartografia como método da pesquisa-in tervenção. In Eduardo Passos, Virgínia Kastrup, & L. Escóssia (Orgs.), Pistas do método da cartografia: Pesquisa-intervenção e produção de subjetividade (pp. 17-31). Sulina. Política Nacional de Assistência Social - PNAS. (2004). Norma Operacional Básica. MDS. Reinach, Sofia (2021). A violência contra crianças e adolescentes na pandemia: análise do perfil das vítimas. In Fórum Brasileiro de Segurança Pública. Anuário brasileiro de segurança pública 2021 (pp. 226-239). FBSP. Passos, Eduardo & Barros, Regina B. (2015). A cartografia como método da pesquisa-in tervenção. In Eduardo Passos, Virgínia Kastrup, & L. Escóssia (Orgs.), Pistas do método da cartografia: Pesquisa-intervenção e produção de subjetividade (pp. 17-31). Sulina. Política Nacional de Assistência Social - PNAS. (2004). Norma Operacional Básica. MDS. Reinach, Sofia (2021). A violência contra crianças e adolescentes na pandemia: análise do perfil das vítimas. In Fórum Brasileiro de Segurança Pública. Anuário brasileiro de segurança pública 2021 (pp. 226-239). FBSP. Política Nacional de Assistência Social - PNAS. (2004). Norma Operacional Básica. MDS. Reinach, Sofia (2021). A violência contra crianças e adolescentes na pandemia: análise do perfil das vítimas. In Fórum Brasileiro de Segurança Pública. Anuário brasileiro de segurança pública 2021 (pp. 226-239). FBSP. Política Nacional de Assistência Social - PNAS. (2004). Norma Operacional Básica. MDS. Reinach, Sofia (2021). A violência contra crianças e adolescentes na pandemia: análise do perfil das vítimas. In Fórum Brasileiro de Segurança Pública. Anuário brasileiro de segurança pública 2021 (pp. 226-239). FBSP. ISSN 1807-0310 13 ESCUTAÇÃO ESCUTAÇÃO Rosa, João G. (1967/2009). Tutaméia (Terceiras estórias). Nova Fronteira. Santos, Boaventura S. (2020). A Cruel Pedagogia do Vírus. Almedina. Schucman, Lia V. (2012). Entre o “encardido”, o “branco” e o “branquíssimo”: raça, hierar quia e poder na construção da branquitude paulistana [Tese de Doutorado, Programa de Pós -graduação em Psicologia, Instituto de Psicologia da Universidade de São Paulo, São Paulo/SP]. Serfaty, Gabriela (2016). Por uma escuta menor [Dissertação de Mestrado, Programa de Pós -graduação em Psicologia Clínica, Pontifícia Universidade Católica de São Paulo, São Paulo/SP]. Vicentin, Maria C. G. (2020). “Criançar o descriançável”: a transicionalidade da infância e o paradoxo da proteção-liberdade. Revista ClimaCom, Devir Criança \| pesquisa – ensaio, 7(18). http://climacom.mudancasclimaticas.net.br/wp-content/uploads/2020/09/ENSAIO-4-Crian-- ar-o-descrian----vel.pdf LETÍCIA EICHHERR https://orcid.org/0000-0002-6840-9342 Mestra em Psicologia Social e Institucional pela Universidade Federal do Rio Grande do Sul (PPGPSI/UFRGS). E-mail: leticiae.psico@gmail.com https://orcid.org/0000-0002-1850-3023 E-mail: lilian.rodrigues.cruz@gmail.com Histórico Submissão: 11/5/2022 Revisão: 27/10/2022 Aceite: 27/10/2022 Contribuição dos autores Concepção: LME, LRC Coleta de dados: LME Análise de dados: LME, LRC Elaboração do manuscrito: LME Revisões de conteúdo intelectual importante: LME, LRC Aprovação final do manuscrito: LME, LRC. Financiamento Bolsa da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) durante 19 meses do mestrado. Aprovação, ética e consenti mento Pesquisa apreciada e aprovada pelo Comitê de Ética em Pesquisa do Instituto de Psicologia da Universidade Federal do Rio Grande do Sul, sob o Certificado de Apresentação de Apreciação Ética (CAEE) número 38494920.7.0000.5334. 14 ISSN 1807-0310	46,762
https://openalex.org/W2588625604	OpenAlex	Open Science	CC-By	2,017	Energy Consumption Analysis for Concrete Residences—A Baseline Study in Taiwan	Kuo-Liang Lin	English	Spoken	8,517	15,271	Energy Consumption Analysis for Concrete Residences—A Baseline Study in Taiwan Kuo-Liang Lin, Ming-Young Jan and Chien-Sen Liao * Department of Civil and Ecological Engineering, I-Shou University, Kaohsiung 84001, Taiwan; kllin@isu.edu.tw (K.-L.L.); myjan@isu.edu.tw (M.-Y.J.) * Correspondence: csliao@isu.edu.tw; Tel.: +886-7-6577711 (ext. 3328); Fax: +886-7-6577461 Academic Editor: Umberto Berardi Kuo-Liang Lin, Ming-Young Jan and Chien-Sen Liao * Department of Civil and Ecological Engineering, I-Shou University, Kaohsiung 84001, Taiwan; kllin@isu.edu.tw (K.-L.L.); myjan@isu.edu.tw (M.-Y.J.) * Correspondence: csliao@isu.edu.tw; Tel.: +886-7-6577711 (ext. 3328); Fax: +886-7-6577461 Academic Editor: Umberto Berardi Kuo-Liang Lin, Ming-Young Jan and Chien-Sen Liao * Kuo-Liang Lin, Ming-Young Jan and Chien-Sen Liao Department of Civil and Ecological Engineering, I-Shou University, Kaohsiung 84001, Taiwan; kllin@isu.edu.tw (K.-L.L.); myjan@isu.edu.tw (M.-Y.J.) * Correspondence: csliao@isu.edu.tw; Tel.: +886-7-6577711 (ext. 3328); Fax: +886-7-6577461 Kuo-Liang Lin, Ming-Young Jan and Chien-Sen Liao Department of Civil and Ecological Engineering, I-Shou University, Kaohsiung 84001, Taiwan; kllin@isu.edu.tw (K.-L.L.); myjan@isu.edu.tw (M.-Y.J.) * Correspondence: csliao@isu.edu.tw; Tel.: +886-7-6577711 (ext. 3328); Fax: +886-7-6577461 Received: 1 December 2016; Accepted: 7 February 2017; Published: 12 February 2017 Received: 1 December 2016; Accepted: 7 February 2017; Published: 12 February 2017 Abstract: Estimating building energy consumption is difﬁcult because it deals with complex interactions among uncertain weather conditions, occupant behaviors, and building characteristics. To facilitate estimation, this study employs a benchmarking methodology to obtain energy baseline for sample buildings. Utilizing a scientiﬁc simulation tool, this study attempts to develop energy consumption baselines of two typical concrete residences in Taiwan, and subsequently allows a simpliﬁed energy consumption prediction process at an early design stage of building development. Using weather data of three metropolitan cities as testbeds, annual energy consumption of two types of modern residences are determined through a series of simulation sessions with different building settings. The impacts of key building characteristics, including building insulation, air tightness, orientation, location, and residence type, are carefully investigated. Sample utility bills are then collected to validate the simulated results, resulting in three adjustment parameters for normalization, including ‘number of residents’, ‘total ﬂoor area’, and ‘air conditioning comfort level’, for justiﬁcation of occupant behaviors in different living conditions. Study results not only provide valuable benchmarking data serving as references for performance evaluation of different energy-saving strategies, but also show how effective extended building insulation, enhanced air tightness, and prudent selection of residence location and orientation can be for successful implementation of building sustainability in tropical and subtropical regions. Keywords: building insulation; energy consumption; baseline; simulation Sustainability 2017, 9, 257; doi:10.3390/su9020257 sustainability sustainability sustainability www.mdpi.com/journal/sustainability where Reqs is region speciﬁc and is set at 13% in north, 15% in middle, and 18% in south. where Reqs is region speciﬁc and is set at 13% in north, 15% in middle, and 18% in south. where Reqs is region speciﬁc and is set at 13% in north, 15% in middle, and 18% in south. In addition to the mandatory building energy standards, the Taiwan government has also proposed a voluntary green building certiﬁcation system called EEWH (Ecology, Energy, Waste Reduction, and Health), as a more aggressive sustainability-driving program. However, voluntary compliance of EEWH has had little commercial success due to limited understanding of the possible beneﬁts frm the program. To implement extra energy-efﬁciency strategies, stakeholders need to estimate energy consumption of each strategy so that they can understand what level of savings can be achieved. However, estimating building energy consumption is difﬁcult because it deals with complex interactions among uncertain weather conditions, occupant behaviors, and building characteristics. 3D simulation tools employing robust physical principles can be used for estimation purposes, but collection and operation of detailed parameters in 3D modeling are time-consuming and cost inefﬁcient at an early stage of building development [15]. Furthermore, due to a lack of precise building and environment parameters and uncertainties of occupant behaviors, discrepancy between data inputs and real situations can easily lead to poor simulation results. To reduce elaborated data inputs, some researchers have developed simpler models for some particular applications. For example, White and Reichmuth [16] proposed to use average monthly temperatures to predict monthly building energy consumption, and the outcome was proven to be more accurate than that of standard simulation procedures using heating and cooling degree days. Nevertheless, when simulation tools are applied, aperformance gap, which is the difference between the predicted energy use and actual measured energy use, always happen and sometimes this gap is too signiﬁcant to be overlooked [17]. To improve simulation accuracy, calibration is suggested in some studies [18]. Repeated calibration to ﬁne tune different inputs can progressively match simulated results with that of a real building [19] but calibration is a tedious and complex process that effectively reveals the demanding efforts to conduct an accurate simulation [20]. In spite of some complications with the data inputs and operations, 3D simulation tools can provide a satisfactory energy performance basis for comparing different design alternatives with proper assumptions [21]. Uar (average U-factor of roof) < 1.0 W/(m2·K) (1) Uaw (average U-factor of wall) < 3.5 W/(m2·K) (2) Req < Reqs (baseline) (3) Uar (average U-factor of roof) < 1.0 W/(m2·K) (1) Uaw (average U-factor of wall) < 3.5 W/(m2·K) (2) Req < Reqs (baseline) (3) (1) (2) (3) (2) (3) 1. Introduction According to the standards, the following three basic code requirements have to be observed: Uar (average U-factor of roof) < 1.0 W/(m2·K) (1) Uaw (average U-factor of wall) < 3.5 W/(m2·K) (2) Req < Reqs (baseline) (3) where Reqs is region speciﬁc and is set at 13% in north 15% in middle and 18% in south 1. Introduction Sustainable development and reduction of energy consumption have been the center of attention in many countries due to the joint effects of climate change, urban heat island effect, rising cost of energy, and environmental concerns [1]. Among all energy consumers, buildings are responsible for over 30% of global energy consumption, and are considered one of the largest energy consumers in modern society [2]. A substantial proportion of building energy consumption comes from the heating, ventilation, and air conditioning systems to enhance indoor thermal comfort [3]. Various modeling studies have predicted that heating, ventilation, and air conditioning system energy demand will rise signiﬁcantly in the near future due to global warming and increased living standards in metropolitan areas [4]. Many researchers have examined regional builiding energy consumptions from different parts of the world, and they have provided valuable information to estimate future sustainable trends [2]. China is the most researched nation for its rising economy and ever-growing energy demands. Case studies with statistical data and/or real-time measurement in Beijing, Shanghai, Guangzhou, Chongqing, Changsha, and Hong Kong have been conducted to identify localized energy consumption patterns and corresponding energy-saving strategies [5–14]. A recent study by Berardi [2] conducts a cross-nation Sustainability 2017, 9, 257; doi:10.3390/su9020257 www.mdpi.com/journal/sustainability 2 of 13 Sustainability 2017, 9, 257 Sustainability 2017, 9, 257 comparison of building energy consumption and shows that the total building energy consumptions in BRIC (Brazil, Russia, India, and China) countries has already surpassed those in developed countries, which creates an urgent need to promote building energy efﬁciency policies in these countries. The study also suggests that appropriate policies coherent with local energy demand patterns are required to achieve substantial building energy reduction. g gy Taiwan started to adopt residential building energy standards in 1997, presenting an ofﬁcial commitment to achieve sustainable development. These national standards are mandatory for a obtaining building permit so that they are profoundly implemented in practice. The standards primarily cover the performance of the building envelope with U-factor requirements in thermal conductivity for the roof and walls, and a Req index (Ratio of Equivalent Transparency Index) for fenestration. where Reqs is region speciﬁc and is set at 13% in north, 15% in middle, and 18% in south. Other than struggling with uncertainties encountered in 3D simulation, a benchmarking system with predetermined baseline consumption data of sample buildings can be employed to predict how a real building with similar characteristics should perform. This kind of benchmarking prediction usually has to be normalized in consideration of the different settings between the sample building and the one being predicted [22,23]. Baseline consumption data of sample buildings can function as an immediate energy comparison indicator, and have the advantage of encouraging poor performers to spot their problematic areas and consequently encourage them to promote efﬁcient use of energy. 3 of 13 energy Sustainability 2017, 9, 257 promote efficient u Although limited in scope, the implementation of energy performance prediction from baseline data is inexpensive, fast, and relatively reliable [24]. Aiming at implementing energy benchmarking to promote building sustainability, this study attempts to develop energy consumption baselines of typical concrete residences in Taiwan with scientific simulation Subsequently the baselines are used to facilitate a simple energy prediction Aiming at implementing energy benchmarking to promote building sustainability, this study attempts to develop energy consumption baselines of typical concrete residences in Taiwan with scientiﬁc simulation. Subsequently, the baselines are used to facilitate a simple energy prediction model at an early design stage of building development. This study is intended to deﬁne what typical modern residences are like and how middle-class occupants behave in Taiwan. Therefore, a residence matching the type of the sample building and the basic assumptions should have a similar energy consumption proﬁle to the baseline. Using a trustworthy simulation engine of a sustainable design software Ecotect Analysis®(Autodesk, San Rafael, CA, USA), and utilizing weather data from three major metropolitan cities of Taiwan as testbeds, two popular types of modern concrete residences, a 5-story single family house and a 12-story high-rise condominium, are modeled. In addition to residence type, other building characteristics considered in the simulation include building insulation, airtightness, orientation, and location. The impacting magnitudes of building characteristics on energy use are analyzed and compared. The goal is to identify key building characteristics that will impact building energy efﬁciency the most for Taiwan’s concrete residences. To validate the simulated results, sample utility bills are then collected and aligned with the baselines. Adjustment parameters for normalizing energy prediction can also be determined with the validation process. scientific simulation. where Reqs is region speciﬁc and is set at 13% in north, 15% in middle, and 18% in south. Subsequently, the baselines are used to facilitate a simple energy prediction model at an early design stage of building development. This study is intended to define what typical modern residences are like and how middle-class occupants behave in Taiwan. Therefore, a residence matching the type of the sample building and the basic assumptions should have a similar energy consumption profile to the baseline. Using a trustworthy simulation engine of a sustainable design software Ecotect Analysis® (Autodesk, San Rafael, California, USA), and utilizing weather data from three major metropolitan cities of Taiwan as testbeds, two popular types of modern concrete residences, a 5-story single family house and a 12-story high-rise condominium, are modeled. In addition to residence type, other building characteristics considered in the simulation include building insulation, airtightness, orientation, and location. The impacting magnitudes of building characteristics on energy use are analyzed and compared. The goal is to identify key building characteristics that will impact building energy efficiency the most for Taiwan’s concrete residences. To validate the simulated results, sample utility bills are then collected and aligned with the baselines. Adjustment parameters for normalizing energy prediction can also be determined with the validation process. 2. Study Area and Model Descriptions 2.2. Model Descriptions Two types of concrete residences are often found in Taiwan: low-rise houses, and high-rise condominiums. Unlike in large countries such as the United States, most suburban low-rise houses in Taiwan do not stand alone due to limited available land and the towering land cost [26]. A popular style of such low-rise residence is a twin-unit type consisting of two identical houses of three to ﬁve stories. The other popular type of residence is high-rise condominiums and most of them are 12 to 14 stories tall with a height of 50 meters or less [27]. Figure 2 shows these two typical modern residence types studied in this research. Case (a) is a twin-unit low-rise house of ﬁve stories and the ﬂoor area of each unit is 397 m2 (794 m2 in total). Case (b) is a high-rise condominium of 12 stories, while each story has two living units (24 units in total) and the ﬂoor area of each unit is 161 m2 (3864 m2 in total). Both cases are based on real projects in the 2010s, targeting middle to upper-class buyers. Floor height for both cases is 3.6 m and the ratio of equivalent transparency (fenestration) is 15%, and their typical ﬂoor plans are shown in Figure 3. This study utilizes Ecotect Analysis 2011 student version as the internal load analysis tool to determine energy consumption, and Ecotect uses the admittance method by the Chartered Institution of Building Services Engineers (CIBSE) for its thermal analysis. Like other thermal analysis methods, the admittance method does not guarantee absolute accuracy of a calculation, but as long as the assumption bases of any comparative calculations are the same and that relative accuracy can be achieved [28]. The purpose of this study takes advantage of the great beneﬁt of the admittance method when it is used as a comparative tool to compare the relative impacts of different design options. For simulation purposes, general assumptions in this study are described as follows: (1) Occupancy and operation setting: the occupancy of each house is set at 6, including grandparents, parents, and a pair of kids. Parents and kids leave home at 8:00 am and come back at 6:00 pm, while grandparents stay at home all day. 2.1. Study Area Situated a Situated at the boundary between the Eurasian Plate and Philippine Sea Plate, Taiwan is an earthquake-prone country (Figure 1). To ﬁght against frequent earthquakes, Taiwan has strict building codes that make concrete the main structural material for modern residential buildings. Concrete, on the other hand, is a poor insulator, which allows the sun’s powerful rays to penetrate through the building envelope and heat up the interior. On a typical summer day in Taiwan, the temperature inside concrete building can easily exceed mid-30 degrees Celsius without air conditioning. earthquake-prone country (Figure 1). To fight against frequent earthquakes, Taiwan has strict building codes that make concrete the main structural material for modern residential buildings. Concrete, on the other hand, is a poor insulator, which allows the sun’s powerful rays to penetrate through the building envelope and heat up the interior. On a typical summer day in Taiwan, the temperature inside concrete building can easily exceed mid-30 degrees Celsius without air conditioning. Figure 1. Study area and three investigated cities of this study: (a) Study area; and (b) three investigated cities. Figure 1. Study area and three investigated cities of this study: (a) Study area; and (b) three investigated cities. Figure 1. Study area and three investigated cities of this study: (a) Study area; and (b) three investigated cities. Figure 1. Study area and three investigated cities of this study: (a) Study area; and (b) three investigated cities. In this study, energy consumption of Taiwan’s three major cities—Taipei, Taichung, and Kaohsiung—are investigated. Taipei and Taichung are located in the subtropical region, and Kaohsiung is located in tropical region. The average temperature of these three cities from 1981 to 2010 is shown in Table 1 [25]. In this study, energy consumption of Taiwan’s three major cities—Taipei, Taichung, and Kaohsiung—are investigated. Taipei and Taichung are located in the subtropical region, and Kaohsiung is located in tropical region. The average temperature of these three cities from 1981 to 2010 is shown in Table 1 [25]. 4 of 13 Sustainability 2017, 9, 257 Table 1. The average temperature (◦C) of three sampled cities from 1981 to 2010. Table 1. The average temperature (◦C) of three sampled cities from 1981 to 2010. 2.1. Study Area Situated a Station January February March April May June July August September October November December Average Taipei 16.1 16.5 18.5 21.9 25.2 27.7 29.6 29.2 27.4 24.5 21.5 17.9 23.0 Taichung 16.6 17.3 19.6 23.1 26.0 27.6 28.6 28.3 27.4 25.2 21.9 18.1 23.3 Kaohsiung 19.3 20.3 22.6 25.4 27.5 28.5 29.2 28.7 28.1 26.7 24.0 20.6 25.1 Weather Data—Major Cities 1981~2010. updated every 10 years, data from Central Weather Bureau 2010 (Taiwan). 2.2. Model Descriptions The occupancy of each unit in the condominium is set at 4 (parents, and a pair of kids), and all four leave home at 8:00 am and come back at 6:00 pm. In both cases, human activity level is in sedentary mode at 70 W; y y (2) Air conditioning: air conditioning is turned on when room temperature is above 26 ◦C in a 24-h setting; ir conditioning: air conditioning is turned on when room temperature is above 26 ◦C in a 4-h setting; (3) Internal gains (values for both lighting, and small power loads per unit ﬂoor area): sensible gain is set at 5 W/m2 (by lighting, and small power loads), and latent gain is 5 W/m2 (by small power loads such as cookers, electric kettle, hot water heaters, etc.) in both cases; (4) Internal design conditions setting: clothing is set at 1.0, relative humidity at 60%, air velocity at 0.5 m/s, and lighting level at 300 lux; and (5) Windows setting: windows remain closed all time. Building orientation is deﬁned as the most important facade of the building, which generally has the most windows and receives the most solar energy. Two orientation options are modeled: ‘facing west’ (denoting the case with largest solar heat reception in summer) and ‘facing north’ (denoting the case with strongest wind chill reception in winter). 5 of 13 4 Sustainability 2017, 9, 257 Sustainability 2017, 9, Sustainability 2017, 9, 257 5 of 14 Figure 2. Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. (b) Facing West or North (a) Facing West or North Figure 2. Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Figure 2. Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. (b) Facing West or North (a) Facing West or North (b) Facing West or North (a) Facing West or North ( ) F i W t N th (a) Facing West or North (b) Facing West or North Figure 2. Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Figure 2. Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Figure 2. 2.2. Model Descriptions Two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. ( ) g Case (a) 5-story low-rise house, floor plan @2F. Figure 3. Cont. Case (a) 5-story low-rise house, floor plan @2F. Figure 3. Cont. Sustainability 2017, 9, 257 6 of 14 Case (b) 12-story high-rise condominium, typical floor plan. Figure 3. Floor Plan for the two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Building orientation is defined as the most important facade of the building, which generally Figure 3. Floor Plan for the two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Case (a) 5-story low-rise house, floor plan @2F. Figure 3. Cont. Case (a) 5-story low-rise house, floor plan @2F. Case (a) 5-story low-rise house, floor plan @2F. Figure 3. Cont. Case (b) 12-story high-rise condominium, typical floor plan. Case (b) 12-story high-rise condominium, typical floor plan. Figure 3. Floor Plan for the two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. Figure 3. Floor Plan for the two typical modern residence types modeled in this study: (a) 5-story low-rise house; and (b) 12-story high-rise condominium. 6 of 13 Sustainability 2017, 9, 257 Sustainability 2017, 9, 257 ‘Typical insulation’ and ‘extended insulation’ are the two options for building insulation. ‘Typical insulation’, the common concrete building envelope in Taiwan [29], uses ceramic tile for wall ﬁnishing, basic roof insulation with lightweight concrete and thin polystyrene foam, and traditional glass for all windows (as shown in Table 2). ‘Typical insulation’ exibits a basic code-compliance thermal conductivity of 3.230 W/m2·k on the wall, 1.00 W/m2·k on the roof, and 6.00 W/m2·k on the windows, respectively. Table 2. ‘Typical insulation’, a minimum insulation setting which meets basic building code requirements, and this represents the most common RC building envelope in Taiwan. Table 2. ‘Typical insulation’, a minimum insulation setting which meets basic building code requirements, and this represents the most common RC building envelope in Taiwan. Layer Name Width Density Sp. 2.2. Model Descriptions Heat U * Wall Composition, U = 3.23 Ceramic Tiles 0.01 2.4 840 1.3 Cement Mortar 0.015 2 800 1.5 Reinforced Concrete 0.15 2.2 880 1.4 Cement Mortar 0.01 2 800 1.5 Roof Composition, U = 1.00 Cement ﬁve-leg tile 0.05 0.7 900 1.5 Polystyrene Foam 0.02 1.04 1130 0.04 Concrete Lightweight 0.05 0.95 656.9 0.8 Asphaltic Felt 0.01 1.02 900 0.11 Cement Mortar 0.02 2 800 1.5 Reinforced Concrete 0.15 2.2 880 1.4 Cement Mortar 0.015 2 800 1.5 Typical Glass, U = 6.0 Glass Standard 0.006 2.3 836.8 1.05 * U-value is the heat transfer coefﬁcient describing how well a building element conducts heat (W/m2·K). Data from Construction and Planning Agency Ministry of the Interior [30]. * U-value is the heat transfer coefﬁcient describing how well a building element conducts heat (W/m2·K). Data from Construction and Planning Agency Ministry of the Interior [30]. ‘Extended insulation’ is an enhanced heat resisting envelope which adds expanded polystyrene (EPS) to the wall and roof, and also uses double-layered Low-E glass for windows. The addition of EPS (with a U-value of 0.04) and Low-E glass reduces the thermal conductivity to 1.07 on the wall, 0.75 W/m2·k on the roof, and 2.5 W/m2·k on the windows. Table 3 shows the material composition of ‘extended insulation’. Table 3. ‘Extended insulation’, a suggested insulation setting with an low overall thermal conductivity which can effectively reduce thermal gain inside the residence. Table 3. ‘Extended insulation’, a suggested insulation setting with an low overall thermal conductivity which can effectively reduce thermal gain inside the residence. Table 3. ‘Extended insulation’, a suggested insulation setting with an low overall thermal conductivity which can effectively reduce thermal gain inside the residence. Layer Name Width Density Sp. Heat U * Wall Composition, U = 1.07 Ceramic Tiles 0.01 2.4 840 1.3 EPS 0.025 21.04 1300 0.04 Cement Mortar 0.015 2 800 1.5 Reinforced Concrete 0.15 2.2 880 1.4 Roof Composition, U = 0.75 Concrete 1-4 Dry 0.05 2.3 800 1.4 PU Block 0.025 1.05 1250 0.028 PU 0.005 1.05 1250 0.05 Cement Mortar 0.015 2 800 1.5 Reinforced Concrete 0.15 2.2 880 1.4 Cement Mortar 0.015 2000 800 1.5 Low-e Glass, U = 2.5 Glass Standard 0.006 2300 836.8 1.046 Air Gap 0.03 1.3 1004 5.56 Glass Standard 0.006 2300 836.8 1.046 * U-value is the heat transfer coefﬁcient describing how well a building element conducts heat (W/m2·K). 3.1. Annual Energy Consumption Creating geometric models, assigning building properties, and applying climate and weather data are the three primary steps required for estimating annual energy consumption using simulation. In this study, two building models are created. Three major sets of building properties are assigned, including two orientations (facing west and facing north), two levels of insulation (typical insulation and extended insulation), and two degrees of air tightness (0.5 ac/h and 3.5 ach). Climate and weather data of three cities (Taipei, Taichung, Kaohsiung) are then applied. The research team has completed a total of 48 different simulation scenarios (2 building types × 2 orientations × 2 insulations × 2 air tightness × 3 cities), and annual energy consumption data of all 48 scenarios is summarized in Table 4. Table 4. Annual Energy Consumption in 48 cases (in KWH). House Condominium Taipei Taichung Kaohsiung Taipei Taichung Kaohsiung Air Leak (3.25 ach) Extended Insulation Facing West 68,879.17 77,094.65 98,015.24 287,970.94 323,584.38 401,207.81 Facing North 68,059.13 75,902.02 96,717.30 286,335.81 320,017.95 398,511.36 Typical Insulation Facing West 81,434.48 * 92,917.36 * 118,164.58 * 357,222.50 * 410,404.90 * 506,015.62 * Facing North 78,978.82 * 89,117.29 * 114,543.31 * 351,622.05 * 399,040.54 * 497,345.66 * Air Tight (0.5 ac/h) Extended Insulation Facing West 38,745.79 42,109.52 53,881.35 184,184.72 190,285.38 236,623.65 Facing North 37,757.78 40,827.71 52,460.78 182,175.39 186,811.36 234,554.69 Typical Insulation Facing West 48,331.52 56,010.93 70,411.45 222,238.40 258,618.91 306,444.61 Facing North 45,650.81 52,038.93 66,643.06 216,380.24 247,091.92 297,598.66 * baseline data for calculating impacting %: Baseline data is the worst energy consumption case in each category in both orientations: Facing West and Facing North and it is can be regarded as the energy demand level of higher living standard in Taiwan. Table 4. Annual Energy Consumption in 48 cases (in KWH). For low-rise house, the maximum annual energy consumption is 118,164.58 KWH, which occurs when the house is facing north, located in Kaohsiung, and with typical insulation and air leak feature of 3.25 ac/h. In contrast, the minimum annual energy consumption is 37,757.78 KWH, when the house is facing west, located in Taipei, and with extended insulation and air tight feature 0.5 ac/h. The most energy-efﬁcient house requires only 31.95% of the least energy-efﬁcient one. In other words, the house at improper location and orientation and without enhanced insulation and air tightness can consume 3.13 times energy of the one with better choices. 2.2. Model Descriptions Data from Construction and Planning Agency Ministry of the Interior [30]. Air tightness is measured in air changes per hour (ac/h) at an indoor-outdoor pressure difference of 50 Pascals, and air change is air ﬂow rate normalized by building interior volume. According to a 7 of 13 Sustainability 2017, 9, 257 case study in Lithuania, the ac/h of normal houses varies [31]. The study showed that ac/h value is between 4.17 and 8.05 when the houses were not properly tightened, and ac/h could be below 0.5 if properly sealed. STR 2.05.01:2005 [32] states that the air change rate (ac/h) has to be three air changes per hour for detached houses without ventilation devices. In Taiwan, no ac/h speciﬁcation has been reinforced, while a study shows an average ac/h (natural ventilation) in a Taiwanese residence is 3–6 [33]. This study assumes that the typical ac/h in Taiwan is 3.25. On the other hand, for energy saving purposes, the ac/h for a properly air tightened residence is set at 0.5. 3.1. Annual Energy Consumption For high-rise condominium, the least energy-efﬁcient type (Kaohsiung, facing west, typical insulation, and air leak) requires 506,015.62 KWH, while the most energy-efﬁcient type (Taipei, facing north, extended insulation, and air tight) requires 182,175.39 KWH, which is only 36.00% of (or 2.78 times more energy-efﬁcient than) the ill-schemed condominium. y gy To examine individual impact of each factor, impacting percentage from the baseline data is calculated and it is deﬁned as the following: Impacting (%) = energy consumption with the examined factor −baseline energy consumption baseline energy consumption Impacting (%) = energy consumption with the examined factor −baseline energy consumption baseline energy consumption Baseline data represents current energy consumption condition, which is always the worst case in each individual category. Table 5 summarizes the baseline data for both house and condominium 8 of 13 Sustainability 2017, 9, 257 in different cities. Comparing the data between house and condominium, it is found that 12-story condominium can achieve around 10% (ranging from 8.01% to 12.03%) energy saving per unit area, and over 44% (ranging from 44.03% to 46.47%) saving per capita when compared with living in house. Totally, each family living in a condominium can save over 62% (ranging from 62.69% to 64.31%) energy annually. It shows that living in the big house is much less energy-efﬁcient than living in the smaller condominium. Table 5. Baseline data for different cities (in KWH). House Condominium Taipei Taichung Kaohsiung Taipei Taichung Kaohsiung Facing West Total Energy 81,434.48 92,917.36 118,164.58 357,222.50 410,404.90 506,015.62 Energy/m2 102.56 117.02 148.82 92.42 (−9.88%) 106.18 (−9.26%) 130.92 (−12.03%) Energy/family 40,717.24 46,458.68 59,082.29 14,884.27 (−63.44%) 17,100.20 (−63.19%) 21,083.98 (−64.31%) Energy/capita 6786.21 7743.11 9847.05 3721.07 (−45.17%) 4275.05 (−44.79%) 5271.00 (−46.47%) Facing North Total Energy 78,978.82 89,117.29 114,543.31 351,622.05 399,040.54 497,345.66 Energy/m2 99.47 112.24 144.26 90.98 (−8.54%) 103.24 (−8.01%) 128.68 (−10.80%) Energy/family 39,489.41 44,558.65 57,271.66 14,650.92 (−62.90%) 16,626.69 (−62.69%) 20,722.74 (−63.82%) Energy/capita 6581.57 7426.44 9545.28 3662.73 (−44.35%) 4156.67 (−44.03%) 5180.68 (−45.73%) House details: 2 families, 6 residents/family, total residents: 12, ﬂoor area: 794 m2 in total or 397 m2/family. Condominium details: 12 stories, 24 families, 4 residents/family, total residents: 96, ﬂoor area: 3865 m2 in total or 161 m2/family. Impacting percentage is calculated by (examined data—baseline data)/baseline data, where house energy consumption is used as baseline. Table 5. Baseline data for different cities (in KWH). House details: 2 families, 6 residents/family, total residents: 12, ﬂoor area: 794 m2 in total or 397 m2/family. Condominium details: 12 stories, 24 families, 4 residents/family, total residents: 96, ﬂoor area: 3865 m2 in total or 161 m2/family. Impacting percentage is calculated by (examined data—baseline data)/baseline data, where house energy consumption is used as baseline. From the impacting percentage calculation on each studied factor, the research team has concluded the following individual impacts of insulation, air tightness, orientation, and location on the two types of residences (as shown in Table 6): 1. Insulation impacts: Annual energy savings from 13.83% to 21.15% are achieved. Average impact is 17.77%. Impacting (%) = energy consumption with the examined factor −baseline energy consumption baseline energy consumption Insulation impacts on the condominium (from 18.57% to 21.15%) appear to be larger than that on the house (from 13.83% to 17.03%). 1. Insulation impacts: Annual energy savings from 13.83% to 21.15% are achieved. Average impact is 17.77%. Insulation impacts on the condominium (from 18.57% to 21.15%) appear to be larger than that on the house (from 13.83% to 17.03%). 2. Air tightness impacts: Annual energy savings from 36.98% to 42.20% are achieved. Average impact is 39.78%. Different from insulation impacts, air tightness impacts on the condominium (from 36.98% to 40.16%) are smaller than that on house (from 39.72% to 42.20%), but the difference is considered not signiﬁcant. 3. Double impacts (insulation + air tightness): Annual energy savings from 48.19% to 54.68% are achieved. Average impact is 52.63%. 4. Orientation impact: Using typical insulation in Taipei as an example ‘facing west’ results in 1.59% (12-story condominium) to 3.11% (5-story house) energy increase. Compared with impacts from insulation and air tightness, orientation impact is small and can almost be neglected. 5. Location impact: Houses (facing north) in Kaohsiung require 45.03% more annual energy than the same house in Taipei. Houses (facing north) in Taichung require 12.84% more annual energy than the same house Taipei. Condominiums (facing north) in Kaohsiung and in Taichung require 41.44% and 13.49% more annual energy than the same condo in Taipei, respectively. Houses and condominiums facing west have similar results. 9 of 13 Sustainability 2017, 9, 257 Table 6. Insulation and air tightness impact on annual energy consumption. Table 6. Insulation and air tightness impact on annual energy consumption. Taipei Taichung Kaohsiung Insulation Impact (ave. −17.77%) House Facing West −15.42% −17.03% −17.05% Facing North −13.83% * −14.83% −15.56% Condominium Facing West −19.39% −21.15% −20.71% Facing North −18.57% −19.80% −19.87% Air Tightness Impact (ave. −39.78%) House Facing West −40.65% −39.72% −40.41% Facing North −42.20% −41.61% −41.82% Condominium Facing West −37.79% −36.98% * −39.44% Facing North −38.46% −38.08% −40.16% Double Impact: Insulation + Air Tightness (ave. −52.63%) House Facing West −52.42% −54.68% ** −54.40% Facing North −52.19% −54.19% −54.20% Condominium Facing West −48.44% −53.63% −53.24% Facing North −48.19% * −53.18% −52.84% * lowest impacting%; ** highest impacting%; Impacting (%) = (examined consumption—baseline consumption)/ baseline consumption. * lowest impacting%; ** highest impacting%; Impacting (%) = (examined consumption—baseline consumption)/ baseline consumption. Among all factors, ‘air tightness’ results in the greatest impact at about 40% energy improvement. Impacting (%) = energy consumption with the examined factor −baseline energy consumption baseline energy consumption On the other hand, envelope insulation, which has been recognized as the most effective energy-saving methodology, results in only less than 18% improvement. This study shows that ‘air tightness’ should be the foremost strategy for home developers. However, building code enforcement of air tightness has been ignored by a lot of countries, including Taiwan. The simulations also show that the impacts of simultaneous implementation of both insulation and air tightness results in 52.63% energy improvement. Further examining the individual impact of insulation and air tightness, which are 17.77% and 39.78% respectively, we have found that the simulated double impact of 52.63% is less than the summation of individual implementation (17.77% + 39.78% = 57.55%). p The same house located in different cities such as Taipei and Kaohsiung results in an over 45% energy consumption variation. This kind of result would not be surprising if the study was on large countries like USA or China which sit across various climate zones with widely ranging temperatures. However, this study is conducted in Taiwan, which is a tiny island with a total area of a mere 36,000 square km. The 45% energy variation is surprisingly high, considering the fact that the two studied cities, Taipei and Kaohsiung, which sit respectively on the north and south of the island, are only separated by 350 km. In fact, the tropic of Cancer (23.5 latitude) cuts through the middle of Taiwan, which separates the entire island into two different climate zones. The northern part of the island belongs to the subtropics, which has warm to hot summers and cool to mild winters with infrequent frost. In contrast, the southern part belongs to tropics, which is warm, hot, and humid in springs, summers, and autumns, with very mild winters. The study shows that the energy consumption depends largely on local climate characteristics and requires regional-speciﬁc investigation to provide accurate predictions. 3.2. Baseline Validation and Adjustment Parameters A sample family’s utility bills ﬁtting the living condition settings in this study are collected to compare with the simulated results. This sample family locates at the 10th ﬂoor a 12-story north-facing condominium in Sungshan district of Taipei City, and has a ﬂoor area of 183 m2. This sample family has three members, including a pair of middle-class parents (double income) and a 10-year-old daughter. The sample family’s utility bills from 2014 through 2016 and the calculated energy consumption indexes are summarized in Table 7. Compared with the sample utility bills, the simulation seems to yield an exaggerated energy consumption number. A possible explanation for such discrepancy is that the sample family usually does not cook at home and does not always demand the highest level of air-conditioning comfort, which could result in some degrees of energy saving. The utility bills also show a constant increase of energy use from 2014 through 2016, and it is possibly because of the 10 of 13 Sustainability 2017, 9, 257 increasing energy demands of the grown-up daughter and the ascending heat island effects in the Taipei metropolitan area as well. Table 7. Annual energy consumption—sample family. 2016 Energy 2015 Energy 2014 Energy November–December 1863 November–December 1858 November–December 1268 September–October 2878 September–October 2477 September–October 1995 July–August 2311 July–August 2406 July–August 1512 May–June 1912 May–June 1708 May–June 1073 March–April 741 March–April 671 March–April 673 January–February 714 January–February 630 January–February 644 Total Energy 10,419 9750 7165 Energy/Capita 3473.00 3250.00 2388.33 Energy/m2 56.93 53.28 39.15 Baseline: Total energy: 14,884 KW, Energy/Capita: 3721 KW, Energy/m2: 92.42 (Taipei, Condominium, Facing West). Past empirical studies have revealed noticeable differences between simulated and measured performances of energy consumption [34,35]. Some studies have also revealed that uncertainties such as occupant behavior, life style and even building construction practices may contribute to the variation in energy consumption in different households, but the extent of their exact inﬂuences are still undetermined [36–38]. The comparison between the simulated data and the actual utility bills shows the same conclusion. To narrow the large performance gap, normalization process is conducted to ﬁt occupant behaviors of the sample family with the simulation settings. Three parameters, including number of residents, ﬂoor area, and desired comfort level are suggested to justify the simulated data for normalization purposes. 3.2. Baseline Validation and Adjustment Parameters A simple normalization approach is employed as suggested in [22,23], so that these three parameters are assumed to be linearly scalable, however exact impacting magnitude may vary from case to case and require further future investigation. These three parameters are explained as follows: 1. Number of residents (Pno_res): 6 residents in house and 4 residents in condominium are assumed in the simulation. Energy consumption is strongly correlated to the number of residents [39]. A linear regression has been used to describe the relationship between occupancy rate and energy consumption [40], suggesting linear relationship a reasonable hypothesis. Hence, a “number of residents” adjustment parameter (Pno_res) in the form of “actual number of residents/6, for house” or “actual number of residents/4, for condominium” is suggested when the number of residents varies. 2. 2. Total ﬂoor area (Pﬂ_area): The initial assumption of the house and condominium area are set at 397 m2 and 161 m2 respectively. These are considered typical settings for upper middle class communities in suburban areas in Taiwan. However, ﬂoor area may vary in different cases. When calculating energy consumption with the admittance method, the internal gains (W/m2), energy use intensity, and lighting intensity (lm/m2, lux) are deﬁned based on ﬂoor area, the resulting energy is directly proportional to ﬂoor area. Energy consumption from air conditioning load due to solar gains is calculated based on the respective sunlit area on wall, roof, and glaze (m2) and the solar intensity (M/m2). Sunlit area on roof is the same as ﬂoor area, and when both fenestration ratio and ﬂoor height are ﬁxed, the envelope area on the wall and glaze is not directly proportional to the ﬂoor area but has strong correlation. This study suggests a linear ‘total ﬂoor area’ adjustment parameter (Pﬂ_area) in the form of “actual ﬂoor area/397, for house” or “actual ﬂoor area/161, for condominium” be asserted in estimation when ﬂoor area varies to simplify the estimate. 3. Air conditioning comfort level (PAC_comf): The simulation assumed air conditioning (AC) is turned on in the entire ﬂoor area when room temperature goes above 26 ◦C in a 24-h setting, no matter whether residents are home or not. This setting results in the maximum comfort, but is not at 3. 3. Annual Energy Consumption = 14,650.92 KWH* (3/4) * (183/161) * (0.8563) = 10,694 KWH (where 14,650.92 KWH is the baseline data retrieved from Table 5). 3.2. Baseline Validation and Adjustment Parameters Air conditioning comfort level (PAC_comf): The simulation assumed air conditioning (AC) is turned on in the entire ﬂoor area when room temperature goes above 26 ◦C in a 24-h setting, no matter whether residents are home or not. This setting results in the maximum comfort, but is not at 11 of 13 Sustainability 2017, 9, 257 all energy efﬁcient. According to the simulation results of such setting, the average daily AC operation time is 7.44 h, and AC accounts for 54.38% of annual energy consumption in concrete residences. That is, when AC is turned off completely, an energy saving of 54.38% can be achieved. However, turning-off AC is not a reasonable scenario in the metropolitan areas in investigated regions, where interior temperature can reach high-30 degrees Celsius in summer. Based on the simulation, a minimum AC energy requirement to secure least comfort level could achieve an energy saving of 43.12% when the AC temperature is turned on at 30 ◦C and in limited hours between 10:00 PM and 4:00 AM and also in limited cooling areas. From the data derived from simulation, this study suggests a four-level air conditioning comfort level setting (PAC_comf) that could be asserted to meet different AC comfort level requirements: (a) for maximum AC comfort, PAC_comf is set at “1”; (b) for moderate AC comfort, PAC_comf is set at “0.8563”(AC on at 27 ◦C and operates for 14 h); (c) for less than moderate AC comfort, PAC_comf is set at “0.7125” (AC on at 28 ◦C and operates for 10 h); and (d) for least AC comfort, PAC_comf is set at “ 0.5688” (1–0.4312) (AC on at 30 ◦C and operates for 6 h). By accompanying with the baseline energy consumption data in Tables 4 and 5, the three proposed adjustment parameters can be used to develop a better prediction of energy consumption appropriate for different living standards. After normalization with these adjustment parameters, the energy consumption can be described as the following equation: Predicted Energy Consumption = (Baseline Data) * (Pno_res) * (Pﬂ_area) * (PAC_comf) For example, we can normalize the energy demand of the sample family (3 members/183 m2/ facing north/condominium/typical insulation/standard air tightness/Taipei/moderate AC comfort) as follows and it shows that the normalization signiﬁcantly close the performance gap: Annual Energy Consumption = 14,650.92 KWH* (3/4) * (183/161) * (0.8563) = 10,694 KWH 4. Conclusions Based on scientiﬁc simulation, this study develops energy consumption baselines for two popular types of modern concrete residences in three major cities of Taiwan. The impacts of residence locations/orientations/types and key building characteristics including envelope insulation and air tightness are analyzed and presented. To validate the baseline, utility bills of a sample middle-class family are collected and compared with the simulated results. For normalization of predicted results, three adjustment parameters (‘number of residents’, ‘total ﬂoor area’, and ‘air conditioning comfort level’) are suggested and proved to narrow the performance gap signiﬁcantly. With the simulated baselines and the three adjustment parameters, an inexpensive, fast, and reliable energy consumption estimate becomes possible. In addition to the demonstration of benchmarking approach for energy prediction, this research has also drawn the following conclusions: (1) Both extended insulation and enhanced airtightness can substantially reduce energy consumption, while air tightness is more effective, reaching an average impact of 39.78%; (2) Buildings facing west always consume more energy than buildings facing north, and this impact is between 1.59% and 3.11%; (3) Location wise, buildings in Kaohsiung are less energy-efﬁcient, consuming 45% more energy than identical buildings in Taipei. As one of the pioneering energy consumption simulation studies in Taiwan, which take into account different locations/orientations and residence types, as well as other key building characteristics, this research has demonstrated that scientiﬁc simulation can provide valuable and accountable data for stakeholders in afﬁrming their energy-reduction implementation decision while pursuing sustainable innovation. During the simulation sessions, this study also found that basic 12 of 13 Sustainability 2017, 9, 257 assumptions such as window open/close, occupancy number, relative humidity, and internal loading are also factors that can affect energy consumption. The exact impacting magnitudes of these assumptions require further detailed investigations in the future. assumptions such as window open/close, occupancy number, relative humidity, and internal loading are also factors that can affect energy consumption. The exact impacting magnitudes of these assumptions require further detailed investigations in the future. Author Contributions: Kuo-Liang Lin managed the study, administered the simulation sessions, and prepared the manuscript. Min-Young Jan conducted the analysis and provided constructive opinions in revision. Chien-Sen Liao edited and revised the manuscript. Conﬂicts of Interest: The authors declare no conﬂict of interest. 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This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).	24,030
https://openalex.org/W1551413167_1	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	7,253	13,776	INVESTIGACIÓN DIDÁCTICA CONOCIMIENTO PREVIO EN FÍSICA DE ESTUDIANTES DE INGENIERÍA Llancaqueo Henríquez, Alfonso1, Caballero Sahelices, M. Concesa2 y Alonqueo Boudon, Paula3 1 Departamento de Ciencias Físicas. Universidad de La Frontera, Temuco, Chile Departamento de Física. Universidad de Burgos, España 3 Departamento de Educación. Universidad de La Frontera, Temuco, Chile allanca@ufro.cl. concesa@ubu.es. palonque@ufro.cl. 2 Resumen. Se presentan resultados de un estudio de conocimiento previo en física, con estudiantes de primer año de ingeniería de la Universidad de La Frontera, Chile, en el marco de la teoría de campos conceptuales de Vergnaud, desarrollada para el estudio de aprendizaje de conceptos cientíﬁcos. El propósito del estudio fue determinar niveles de comprensión de conceptos de física que describan la estructura del conocimiento previo de los estudiantes en un curso inicial de física, a partir del uso de los conceptos a) En situaciones de contexto cotidiano, b) En la comprensión de un texto de divulgación cientíﬁca, y c) En situaciones y problemas de contexto académico. Los resultados obtenidos conﬁrman bajos niveles de compresión de los estudiantes al enfrentar situaciones y lecturas que demandan una conceptualización cientíﬁca aceptable. Se observa un predominio de aspectos procedimentales de las operaciones y escaso uso de signiﬁcados de mayor riqueza conceptual. Palabras clave. Situaciones, estructura del conocimiento previo, aprendizaje, conceptos físicos. Undergraduate students’ previous knowledge of Physics Summary. Some research results of a study on previous knowledge in Physics, carried out with ﬁrst year engineering students at the University of The Frontera, Chile are presented. Vergnaud’s Conceptual Fields Theory is used to discover the way to learn scientiﬁc concepts. The purpose of the study was to determine comprehension levels of concepts of physics, which describe the structure of the previous knowledge students had in an initial course of Physics, from the use of concepts: a) In situations of daily context, b) In the comprehension of a text of scientiﬁc dissemination, and c) In situations and problems of academic context. The obtained results conﬁrm low levels of understanding of the students when facing situations and readings which require a scientiﬁc acceptable conceptualization. A predominance of procedural aspects of the operations and limited use of meanings of major conceptual richness is observed. Keywords. Situations, previous knowledge structure, learning, physical concepts. INTRODUCCIÓN En la perspectiva del mejoramiento de la calidad de la enseñanza de física, de proveer oportunidades reales para el aprendizaje de la física a los estudiantes que ingresan a carreras de ingeniería en la Universidad de La Frontera, Chile, se realizó un estudio de conocimientos en física de los estudiantes al inicio del primer curso de física, con el ﬁn de caracterizar el conocimiento previo, ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2), 205–216 describir niveles de comprensión de los signiﬁcados cientíﬁcos de los conceptos e identiﬁcar disposiciones de aprendizaje. Lo anterior implica indagar sobre cuáles son las representaciones de conocimiento de física de los estudiantes al comenzar su aprendizaje en la universidad. Sin embargo, este acervo no se reﬁere sólo a los conocimientos de física que los estudiantes pudieren 205 INVESTIGACIÓN DIDÁCTICA traer de la enseñanza media, sino que en general, considera comprensiones que están en los estudiantes tanto como efecto de su enseñanza como de sus peculiaridades culturales. De este modo, el sentido de este estudio es más amplio que una investigación orientada sólo a determinar los conocimientos escolares previos. Además, se espera que sus resultados permitan dar cuenta en forma sistematizada del punto de inicio en que se encuentran los estudiantes más avanzados y menos avanzados. Es decir, se trata de establecer el punto de inicio para un proceso de enseñanza que permita avanzar a los estudiantes en el desarrollo conceptual de la física (Moreira, 2000; 2004). supone deberían haber logrado en la enseñanza media. Muchas veces, por diversos motivos, estas habilidades de comprensión y procedimientos no se han logrado. Por ejemplo, es común que muchos estudiantes no puedan explicar determinados fenómenos o resolver problemas, usando conocimientos y procedimientos de la física que se presuponen aprendidos con la enseñanza. Conocer estas habilidades de comprensión y procedimientos es un requisito necesario para que en la universidad se diseñe una enseñanza que impacte efectivamente en la progresividad del aprendizaje signiﬁcativo de los estudiantes (Caballero, 2004; Moreira, 2000). En resumen, el estudio realizado estuvo orientado a hacer explícitos los signiﬁcados y grados de comprensión que los estudiantes tienen al inicio de un curso de física en la universidad. FUNDAMENTACIÓN TEÓRICA La estructura y organización de este estudio considera tres aspectos. El primero se reﬁere a una exploración de la comprensión de los signiﬁcados cientíﬁcos que los estudiantes tienen acerca de un conjunto de conceptos de física enseñados en la educación secundaria. El propósito fue indagar qué representación de conceptos como energía y fuerza tiene un estudiante y cómo comprende estos conceptos en situaciones de contexto cotidiano. La importancia de identiﬁcar estas representaciones consiste, como lo muestran numerosas investigaciones, en que éstas se arraigan poderosamente en la mente de los estudiantes desde una edad muy temprana y, muchas veces, el aprendizaje memorístico de los contenidos no logra desplazarlas durante toda su enseñanza (Driver, Guesne, Tiberghien, 1985; Driver, Squires, Rushworth y Wood-Robinson, 1999; Pozo y Gómez Crepo, 1998; Pozo, 1999; Duit, 2003). Estas representaciones constituyen obstáculos poderosos que diﬁcultan el aprendizaje de los conceptos y la comprensión de fenómenos descritos por la física. Por lo tanto, si se considera que todo aprendizaje contiene comprensiones culturales para los estudiantes, el conocimiento de éstas ayudaría a un profesor a tener claridad de estas comprensiones para sus estudiantes; esto facilitaría el diseño de actividades de aprendizaje que den a los estudiantes posibilidades para acceder a un conocimiento de la física distinto al contenido en su saber cotidiano. El segundo aspecto investigado se reﬁere a la compresión de signiﬁcados cientíﬁcos de conceptos de física expresados en un texto. Se trata de averiguar si los estudiantes comprenden el lenguaje de la física, si pueden comunicar y explicar los fenómenos usando este lenguaje, para así conocer sus comprensiones y lo que pueden escribir y explicar. Este planteamiento obecede a que, en muchas ocasiones, los estudiantes no comprenden los signiﬁcados y estructuras simbólicas contenidos en el lenguaje de la física, lo cual obstaculiza el logro de aprendizajes signiﬁcativos de la física (Vergnaud, 1998). El tercer aspecto se reﬁere a una exploración de la comprensión de los signiﬁcados cientíﬁcos de los conceptos por parte de los estudiantes, al enfrentar situaciones y problemas de contexto académico. El propósito es conocer la comprensión de signiﬁcados y habilidades procedimentales de los estudiantes, que se 206 Como referencial teórico para determinar los grados de comprensión de conceptos de física que caracterizan el conocimiento previo de los estudiantes, se adoptó usar la teoría de campos conceptuales de Vergnaud. Esta teoría tiene por premisa que el conocimiento se encuentra organizado en campos conceptuales de los que los sujetos se apropian a lo largo del tiempo. Los campos conceptuales se deﬁnen como conjuntos informales y heterogéneos de situaciones y problemas, que para su análisis y tratamiento requieren diversas clases de conceptos, representaciones simbólicas, operaciones de pensamiento y procedimientos que se conectan entre sí durante su aprendizaje (Vergnaud, 1981; 1990). Esta teoría permite analizar la relación entre los conocimientos explícitos, de los conceptos construidos por las disciplinas, e implícitos del comportamiento de los sujetos al enfrentar situaciones. Además, permite analizar las relaciones entre los signiﬁcados y signiﬁcantes de un concepto (Vergnaud, 1990). En este enfoque, una situación es entendida como una combinación de tareas y problemas que determinan los procesos cognitivos y las respuestas de un sujeto (Vergnaud, 1990); éste, a través de su acción y dominio progresivo de situaciones, adquiere los campos conceptuales que modelan su conocimiento. Así, el papel de la enseñanza es fundamental, pues a través de ésta se intenta establecer una interacción entre los esquemas de los estudiantes y los campos conceptuales de la física descritos cientíﬁcamente. Desde el punto de vista de Vergnaud (1996), el aprendizaje de los conceptos cientíﬁcos se hace posible a través del proceso de mediación planteado por la teoría de Vygotsky, para quien la actividad humana está mediatizada, por una parte, por la interacción social y por otra, por el uso de signos e instrumentos, siendo el más importante de éstos el lenguaje. El desarrollo de los procesos mentales superiores depende de la descontextualización, y el lenguaje permite apartarse de referentes concretos. De este modo, la mediación hace posible el desarrollo de tales procesos (Riviere, 1994). En este sentido, Vygotsky (1995) plantea que los conceptos cientíﬁcos se distinguen de los cotidianos en que los primeros tienen un valor mayor de generalidad y forman sistemas integrados. Se transmiten en instituciones educativas a través de la acción intencionada del docente ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) INVESTIGACIÓN DIDÁCTICA y son un objeto de comunicación que se expresa a través de medios lingüísticos y otros medios simbólicos, como por ejemplo el espacio gráﬁco. Vergnaud destaca la importancia de esta distinción entre conceptos cotidianos y cientíﬁcos, pues la expresión de estos últimos permite cambiar el estatuto cognitivo de los invariantes operatorios, de las reglas de acción, de los objetivos, de las anticipaciones y de las inferencias contenidas en los esquemas (Vergnaud, 1996). En este marco, el docente tiene un papel de mediador, planteando situaciones y problemas a través de instrumentos de mediación semiótica como el lenguaje y los signos (Vergnaud, 1998). La relación existente entre aprendizaje y desarrollo cognitivo se explica a través del concepto de zona de desarrollo próximo (ZDP). Vygotsky planteaba que el aprendizaje se produce sólo cuando los instrumentos, signos, símbolos y normas de los compañeros de interacción pueden ser incorporados en función del nivel de desarrollo previo, es decir, el aprendizaje depende también del desarrollo potencial del sujeto (Vygotsky, 1989). En este sentido, las habilidades de resolución de problemas que se aplican a la realización de tareas pueden ubicarse en tres categorías: a) aquellas realizadas independientemente por el estudiante, referidas a funciones que ya han madurado; b) aquellas que no pueden realizarse aun con ayuda, y c) aquellas que caen dentro de estos dos extremos, las que pueden realizarse con la ayuda de otros, pues se encuentran en un proceso de maduración (Rivière, 1994). De acuerdo con una interpretación de la teoría de Vergnaud, un campo conceptual de conceptos de física estaría compuesto por: C = {S, I; R}. Donde, S = {FF }: es un conjunto de situaciones que incluye fenómenos y problemas físicos (FF) que dan sentido a los conceptos en algún dominio especíﬁco de la física, como la mecánica, electromagnetismo u otros. I = {I(FF) ∪ I(OM)}: es un conjunto de invariantes operatorios físicos I(FF) y matemáticos I(OM), cientíﬁcamente aceptados, que se aplican a las situaciones, es decir, se aplican tanto a los problemas y fenómenos físicos FF como a los objetos matemáticos OM. R = {R[(FF) ∪ I(FF) ∪ I(OM)]}: es un conjunto de representaciones simbólicas y pictóricas usadas en los problemas y situaciones FF, más las representaciones simbólicas de los invariantes operatorios físicos I(FF) y matemáticos I(OM), o sea, las representaciones de los principios y leyes físicas, junto con las operaciones matemáticas y sus propiedades (Llancaqueo, Caballero y Moreira, 2003). Por lo tanto, aplicar la teoría de los campos conceptuales a investigaciones sobre aprendizaje de conceptos de física supone enfrentar a los participantes a situaciones y problemas presentados en diferentes formatos representacionales (imágenes, gráﬁcas, textos), para investigar los invariantes operatorios usados para comprender y explicar las representaciones simbólicas que ayudan a la conceptualización de los signiﬁcados de los conceptos (Moreira, 2002; Greca y Moreira, 2002). En consecuencia, el presente estudio busca establecer la zona de desarrollo real (ZDR) o punto de inicio para un proceso de enseñanza en la ZDP que permita avanzar en el desarrollo y aprendizaje signiﬁcativo de los estudiantes y en acuerdo con las demandas del currículo. METODOLOGÍA Participantes La muestra estuvo formada por 198 estudiantes de las asignaturas de Física General e Introducción a la Física de carreras de ingeniería de la Universidad de La Frontera, Chile, año 2004, distribuidos como se describe en la tabla 1. Del total de estudiantes participantes, 157 son hombres y 41 mujeres, con edades entre los 17 y 22 años. Las mayores frecuencias de edad corresponden a 18 años (65 %), 19 años (15 %) y 17 años (12 %). Por otra parte, el 81 % de los participantes egresó de la enseñanza media el año anterior y el 64 % de los estudiantes mencionó haber tomado un curso electivo de física. Además, en la tabla 1 se presentan los puntajes de selección máximos, mínimos y promedios en las carreras participantes. Tabla 1 Distribución de estudiantes y puntajes de ingreso en las carreras de ingeniería participantes en el estudio. FÍSICA GENERAL CARRERAS INT. FÍSICA PUNTAJE DE SELECCIÓN Máximo Mínimo Promedio Ingeniería Civil Industrial, Bioprocesos (ICIB) 29 823,20 555,95 652,80 Ingeniería Civil Industrial, Informática (ICII) 76 771,86 599,29 653,04 Ingeniería Civil Electrónica (ICE) 34 730,38 515,34 607,17 Ingeniería Matemática (IM) 29 667,59 486,20 562,15 693,15 489,62 562,41 Ingeniería Ejecución Electrónica (IEEa) 30 Total participantes 168 30 Fuente: Oﬁcina de Admisión y Matrícula UFRO, 2004. ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) 207 INVESTIGACIÓN DIDÁCTICA lecturas. La descripción de las situaciones y las lecturas se presentan en la tabla 2. Instrumento Considerando que no se disponía de un instrumento para obtener datos que permitieran analizar y caracterizar el conocimiento previo y los grados de comprensión de los signiﬁcados cientíﬁcos de los estudiantes, en el sentido descrito por la teoría de Vergnaud, se procedió a elaborar un instrumento ad-hoc para la realización de este estudio. Se seleccionaron dos lecturas tomadas de textos de divulgación cientíﬁca y se diseñaron una serie de situaciones con tareas y problemas de lápiz y papel, de las cuales se seleccionaron diez para la construcción deﬁnitiva de un cuestionario de lápiz y papel (Anexo 1). En el cuestionario se presentan situaciones con un formato diverso, con preguntas de respuesta abierta, de modo que los datos obtenidos permitan inferir aspectos de las representaciones y signiﬁcados de los conceptos usados por los estudiantes para dar cuenta de las situaciones y Procedimiento El cuestionario se administró colectivamente en una sala de clase a grupos de estudiantes de ingeniería de los cursos de Física General e Introducción a la Física de la Universidad de La Frontera. La aplicación tuvo una duración de 60 minutos aproximadamente. Análisis de datos Los datos obtenidos con la aplicación del instrumento fueron sometidos a un proceso de codiﬁcación, y se establecieron cuatro categorías de desempeño que reﬂejaran Tabla 2 Contenidos de las situaciones y lecturas del instrumento. Uso de conceptos en situaciones de contexto cotidiano Situación Conceptos Descripción Pregunta S1 Fuerza y movimiento Uso del concepto de fuerza para explicar un movimiento P1 y P2 S2 Fuerza y energía Signiﬁcado de los conceptos de fuerza y energía P3 y P4 S3 Ondas electromagnéticas Comprensión de un modelo que explique la comunicación inalámbrica P5 Uso de conceptos en la comprensión de un texto Lectura Conceptos Descripción Pregunta L1 Densidad, masa y volumen Uso y comprensión de los conceptos de densidad, masa y volumen en el contexto de una situación presentada en un texto L2 Conservación de la energía Comprensión de analogías para una magnitud conservada P6 , P7 y P8 P9, P10 y P11 Uso de conceptos en situaciones y problemas de contexto académico Situación Descripción Pregunta S4 Comprensión del concepto de fuerza normal y sus signiﬁcados para explicar el movimiento curvilíneo de un cuerpo S5 Fuerza de gravedad, masa y energía Comprensión de los conceptos de fuerza de gravedad, masa, aceleración y energía en una situación de caída libre S6 Centro de gravedad y equilibrio Comprensión de conceptos y leyes de la mecánica para describir y explicar una situación de equilibrio P15 S7 Reﬂexión de la luz y visión Comprensión de un modelo para la visión basado en la reﬂexión de la luz P16 S8 Equilibrio térmico Representación y signiﬁcados de los conceptos de temperatura y equilibrio térmico P17 S9 Transferencia calor Comprensión y signiﬁcados del proceso de transferencia del calor por conducción P18 Principios de Newton Comprensión principios de Newton para explicar el estado de reposo o movimiento de un cuerpo Comprensión del principio de acción y reacción para describir y explicar las fuerzas que actúan sobre un cuerpo Comprensión de la ecuación de movimiento para explicar y predecir la evolución de un movimiento S10 208 Conceptos Fuerza normal y movimiento P12 P13 y P14 P19, P20, P21 y P22 ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) INVESTIGACIÓN DIDÁCTICA Para investigar la calidad de los puntajes, se realizó un análisis de coherencia mediante el coeﬁciente α (Cronbach, 1967), que mide la parte estable, conﬁable y común de los ítems. La síntesis del análisis de ﬁabilidad se presenta en la tabla 3. Por lo tanto, dado que con este instrumento se intenta medir aspectos de las representaciones de los estudiantes largamente implícitos, se estima que el valor α = 0,80 indica que los puntajes generados son estadísticamente conﬁables. La validación de contenido del cuestionario se realizó mediante un análisis de contenidos de las situaciones y lecturas y de los supuestos que le dieron origen (Silveira, 1981). El instrumento se sometió al juicio de cinco profesores del curso de física general, procurando lograr un consenso de los contenidos del instrumento (Silveira, 1993). Sección 1: Situaciones de contexto cotidiano (SCC) 20 puntos, Sección 2: Lecturas de textos científicos (LTC) 24 puntos y Sección 3: Situaciones y problemas de contexto académico (SCA) 44 puntos, total 88 puntos. Figura 1 Distribución puntaje promedio en cada sección. 20 16,0 Puntaje promedio de manera jerárquica el uso de los signiﬁcados usados por los estudiantes en su interacción con los contenidos e información de las situaciones y lecturas. La codiﬁcación de las respuestas se realizó mediante una escala de puntaje de desempeño de 0 a 4, donde el valor 0 corresponde a no contesta; 1 a un desempeño de desacuerdo con signiﬁcados cientíﬁcos de los conceptos; 2 en acuerdo parcial mínimo con signiﬁcados cientíﬁcos de los conceptos; 3 en acuerdo parcial y 4 de acuerdo con signiﬁcados cientíﬁcos. La puntuación máxima posible fue 88, y en el anexo 2 se presentan, a modo de ejemplo, criterios de corrección usados para evaluar el desempeño de los estudiantes en las situaciones y lecturas del instrumento. La justiﬁcación de estos criterios es poder identiﬁcar la disponibilidad de signiﬁcados cientíﬁcos en la estructura conceptual de los estudiantes, para aproximarse así a una determinación de los grados de comprensión de su conocimiento previo. 9,8 10 5 0 SCC LTC Figura 2 Desempeño promedio por carrera en cada sección. Sección 1: SCC Sección 3: SCA 0,80 0 ,6 6 0 ,6 3 De toda la información obtenida con la aplicación del instrumento, en la ﬁgura 1 se presenta el puntaje promedio obtenido por la población de estudiantes en cada sección; el puntaje máximo posible de obtener en cada sección es: Sección 2: LTC 1,00 0,60 Descripción del puntaje promedio de desempeño SCA Los valores de puntaje promedio de la población de estudiantes en cada sección, normalizados en una escala de 0,00 a 1,00, representan desempeños de 0,49 en SCC; 0,61 en LTC y 0,36 en SCA. Según estos resultados, el mayor desempeño de los estudiantes es en la sección de comprensión de textos de divulgación cientíﬁca, seguido del desempeño en la sección de situaciones de contexto cotidiano, y el menor en la sección de situaciones y problemas de contexto académico. Este mismo patrón de comportamiento para el desempeño por sección, se observa en todas las carreras, como se muestra en la ﬁgura 2. Además, las carreras ICIB e ICII se destacan por tener valores de desempeño mayor al desempeño promedio de la población de estudiantes en todas las secciones del estudio. RESULTADOS Los resultados obtenidos se presentan en dos partes. La primera parte se reﬁere a la descripción del puntaje obtenido por los estudiantes en las diferentes secciones del instrumento. La segunda parte describe y presenta la asignación de niveles de comprensión de los conocimientos de física de los participantes. 14,7 15 0 ,59 0 ,55 0 ,51 0 ,52 0 ,51 0 ,50 0 ,4 4 0 ,3 7 0,40 0 ,3 8 0 ,3 4 0 ,4 0 0 ,3 4 0 ,2 5 0,20 0,00 ICIB ICII ICE IM IEEa Tabla 3 Síntesis del análisis de fiabilidad. PUNTAJE TOTAL MEDIA PUNTAJE TOTAL DESVIACIÓN TÍPICA Nº ÍTEM COEFICIENTE α 88 40,55 11,76 22 0,80 ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) 209 INVESTIGACIÓN DIDÁCTICA Por otra parte, si se comparan los desempeños por carrera respecto al desempeño promedio de la población en cada sección, se encuentra que tres carreras ICIB, ICII e ICE tienen desempeños superiores al promedio de la población en la Sección 1. Esto mismo se cumple en la Sección 2 con las carreras ICIB e ICII; y en la Sección 3 con las tres carreras ICIB, ICII y IEE. Niveles de comprensión Con el propósito de determinar grados de compresión de los conceptos de física que describan la estructura del conocimiento previo de los estudiantes y caracterizar el punto de partida de la enseñanza, se deﬁnieron cuatro niveles de comprensión. Nivel 1: Incomprensión, Nivel 2: Comprensión incipiente, Nivel 3: Comprensión parcial y Nivel 4: Comprensión. La descripción de cada nivel se presenta en la tabla 4. Para la asignación de los estudiantes en los niveles de comprensión, se analizó el desempeño obtenido por cada estudiante en cada sección de este estudio. Así, un estudiante con desempeño en el rango de 1,00 a 0,75 se ubica en el nivel de comprensión completa de signiﬁcados de los conceptos. Un desempeño entre 0,74 y 0,50 se interpreta como un nivel de comprensión parcial de signiﬁcados. Un desempeño entre 0,49 y 0,25 correspondería a un nivel de comprensión de signiﬁcados incipiente. Por último un desempeño menor a 0,25 correspondería a un nivel de incomprensión de signiﬁcados por parte de un estudiante. La distribución del número de estudiantes por carrera en cada uno de los niveles de comprensión se muestra en la tabla 5. Además, en la ﬁgura 3 se presenta la distribución porcentual de estudiantes por carrera en los cuatro niveles de comprensión. Un análisis preliminar de estos resultados muestra que la mayoría de los estudiantes se ubica en los niveles intermedios de comprensión, con 91 estudiantes en el nivel de comprensión incipiente y 89 estudiantes en el nivel de comprensión parcial. Por otra parte, 13 estudiantes se ubican en el nivel de incomprensión, y sólo 5 estudiantes en el nivel más alto de comprensión. En general, estos Tabla 4 Descripción de niveles de comprensión. NIVEL N1 N2 N3 N4 DESCRIPCIÓN Incomprensión. Predominio o ausencia de esquemas con signiﬁcados que evidencian una comprensión de los conceptos de física. No contesta o escribe respuestas irrelevantes, o no utiliza los conceptos pertinentes en sus explicaciones, o confunde los términos estableciendo vínculos en desacuerdo con signiﬁcados cientíﬁcos de los conceptos. Comprensión incipiente. Reconocimiento sin explicación, o explicitación parcial de signiﬁcados de los conceptos cientíﬁcamente aceptados. No maniﬁesta una conceptualización cientíﬁca aceptable de la física, sólo reconoce las situaciones. Si bien intenta enfrentarlas y describirlas mediante el uso de conceptos y leyes de la física, aún no explica o usa operaciones y representaciones simbólicas que reﬂejen una comprensión de signiﬁcados cientíﬁcos de los conceptos. Comprensión parcial. Transición entre un reconocimiento y la signiﬁcación parcial de los conceptos de la física con aplicación a situaciones y problemas. Reconoce situaciones a las que aplica conceptos y leyes de la física. En las explicaciones se reﬂeja organización y comprensión de signiﬁcados de los conceptos, operaciones y sus representaciones simbólicas, pero aún no logra conectarlos completamente. Comprensión. Aprehensión de la mayoría de los conceptos para el nivel de instrucción. Maniﬁesta comprensión y explicitación de los signiﬁcados de los conceptos de física al enfrentar y describir las situaciones, también usa apropiadamente las operaciones, representaciones simbólicas y propiedades de los conceptos en situaciones y problemas. Tabla 5 Distribución del número de estudiantes según niveles de comprensión. N1 INCOMPRENSIÓN N2 COMPRENSIÓN INCIPIENTE N3 COMPRENSIÓN PARCIAL N4 COMPRENSIÓN TOTAL ICIB 0 5 21 3 29 ICII 2 33 39 2 76 ICE 3 19 12 0 34 IM 8 15 6 0 29 IEEa 0 19 11 0 30 Total 13 91 89 5 198 CARRERA 210 ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) INVESTIGACIÓN DIDÁCTICA resultados estarían mostrando que la mayoría de los estudiantes presenta un conocimiento previo que se caracteriza por un predominio de signiﬁcados de los conceptos en desacuerdo con sus signiﬁcados cientíﬁcos. Figura 3 Distribución porcentual de estudiantes por carrera en los niveles de comprensión. N1: incomprensión N2: comprensión incipiente N3: comprensión parcial N4: comprensión 100% 80% 72% 63% 60% 56% 51% 52% 43% 37% 35% 40% 28% 21% 17% 20% 10% 9% 3% 0% 3% 0% 0% 0% 0% 0% ICIB ICII ICE IM IEEa DISCUSIÓN Y CONCLUSIÓN Se observa que la mayoría de los estudiantes participantes en este estudio presenta una ausencia notoria de conocimientos en física para enfrentar las situaciones y lecturas propuestas en el cuestionario. De acuerdo con los resultados presentados en la tabla 5, se deduce que el 97% de los estudiantes se ubica en los niveles más bajos de comprensión (niveles N1, N2 y N3) deﬁnidos para este estudio, donde el nivel N3 corresponde tan sólo a un nivel de compresión parcial de signiﬁcados cientíﬁcos de un conjunto importante de conceptos y leyes de la física. Los bajos niveles de comprensión se maniﬁestan en todas las carreras (Fig. 3), con un predominio de estudiantes en los niveles de compresión incipiente y parcial. Así en las carreras ICE, IM y IEEa el porcentaje de estudiantes en el nivel incipiente es de entre un 56 % y 63 %; en el nivel de comprensión parcial es de entre 21 % y 37 %, y ningún estudiante alcanza el nivel de comprensión. Sólo dos carreras ICIB e ICII, presentan una distribución de estudiantes con predominio en los niveles de comprensión más altos. En efecto, la distribución observada para estas carreras es de entre el 17 % y 43 % de sus estudiantes en el nivel incipiente, de entre el 72 % y el 51 % en el nivel aceptable, y de entre el 10 % y 3 % en el nivel de comprensión, que corresponde a cinco estudiantes del total de la muestra. Además, llama la atención que 13 estudiantes pertenecientes a las carreras de ICII, ICE e IM, que representan el 7 % del total de la muestra, se ubiquen en el nivel de incomprensión, que corresponde a un nivel de conocimiento y uso de conceptos en desacuerdo con sus signiﬁcados cientíﬁcos. Si se analizan los puntajes promedios de desempeño en cada sección, mostrados en la ﬁgura 1, se observa que el ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) mejor desempeño está en la sección de LTC (0,61), que explora la comprensión lectora de textos cientíﬁcos; le sigue el desempeño en la sección SCC (0,49), que explora la comprensión y uso de conceptos de física en situaciones de contexto cotidiano. El desempeño más bajo está en la sección SCA (0,36), que explora la compresión y uso de conceptos de física en situaciones de contexto académico. Sin embargo, si además del análisis descrito en el párrafo anterior se analiza el desempeño promedio por carrera en cada una de las secciones (Fig. 2), se observa que en todas las carreras se repite este patrón de comportamiento del desempeño por sección. Llama la atención que el mayor desempeño en la Sección SCA corresponda a la carrera de IEEa (0,40), que a su vez es una de las carreras con menor puntaje de selección (Tabla 1). En cambio el mayor desempeño en la sección LTC corresponde a la carrera de ICII (0,66) y el mayor desempeño en la sección SCC corresponde a la carrera de ICIB (0,55), que son las carreras con mayor puntaje de selección (Tabla 1). Este mejor desempeño de los estudiantes de las carreras ICII y ICIB en situaciones cotidianas y comprensión de lectura podría estar indicando que las representaciones (esquemas) de los conceptos de física de estudiantes con mayor puntaje de selección se caracterizarían por una mayor riqueza conceptual con una mejor discriminación de signiﬁcados (Moreira, 2000; Caballero, 2003). En cambio, los esquemas de los estudiantes de carreras de menor puntaje de selección y con mejor desempeño en situaciones de contexto académico, como la carrera IEEa, se caracterizarían por una representación conceptual más restringida, con predominio de aspectos procedimentales (Vergnaud, 1998). La baja comprensión conceptual y la baja discriminación de significados de los conceptos usados por los estudiantes se observan con más nitidez al analizar la frecuencia del desempeño en las preguntas P4, P9, P10, P11 y P14 del cuestionario (Anexo 1), que se refieren al concepto de energía, como se muestra en la tabla 6. En la tabla 6 se observa que el nivel de desempeño máximo (de acuerdo con signiﬁcados cientíﬁcos) es alcanzado por muy pocos estudiantes en las preguntas P9 y P14, las cuales demandan un uso de signiﬁcados del concepto de energía de mayor riqueza conceptual. En efecto, sólo 15 estudiantes (7,6%) alcanzan este nivel en la pregunta P9, la cual demanda la construcción de una analogía que explique la conservación de la energía; y 13 estudiantes (6,6%) alcanzan este mismo nivel en la pregunta P14, que demanda identiﬁcar y aplicar el concepto de energía y su conservación en la resolución de un problema. En cambio, se observa que un número mayor de estudiantes alcanza el nivel de desempeño máximo en las preguntas P4, P10 y P11, que demandan una comprensión menor de signiﬁcados, como son, la construcción de frases que incluyan el concepto de energía y el cálculo de cantidades asociadas a una magnitud conservada a partir de una expresión matemática dada. 211 INVESTIGACIÓN DIDÁCTICA Tabla 6 Frecuencia de desempeño en preguntas del concepto de energía. FRECUENCIA NIVEL DE DESEMPEÑO Desacuerdo con signiﬁcados cientíﬁcos P4 (SCC) P9 (LTC) P10 (LTC) P11 (LTC) P14 (SCA) 36 42 33 34 47 Acuerdo parcial mínimo con signiﬁcados cientíﬁcos 21 37 5 6 104 Acuerdo parcial con signiﬁcados cientíﬁcos 46 19 3 1 27 Acuerdo con signiﬁcados cientíﬁcos 77 15 92 113 13 No contesta Total estudiantes 18 85 65 44 7 198 198 198 198 198 Esta falta de una compresión más profunda del concepto de energía por parte de los estudiantes, manifestada en el desempeño de las preguntas P9 y P14, es coherente con el número de estudiantes que no contestan la pregunta P9 (85) y con el alto número de estudiantes en el nivel de desempeño de acuerdo parcial mínimo con signiﬁcados cientíﬁcos (104) (Tabla 4). conceptual y metodológico para el desarrollo de diseños de enseñanza de conceptos de física en la ZDP que atiendan la diversidad de los estudiantes. Además, esto facilitaría el desarrollo de acciones que permitan organizar y optimizar el proceso de enseñanza-aprendizaje para asegurar oportunidades para que todos puedan aprender (Allal y Pelgrims, 2000). De acuerdo con lo anterior, se puede concluir que los grados de comprensión que caracterizan el conocimiento previo de los estudiantes permiten identiﬁcar vínculos entre la estructura de las representaciones de los estudiantes y la estructura de signiﬁcados, que éstos utilizan al enfrentar situaciones y problemas que involucran una conceptualización cientíﬁcamente aceptable de un conjunto importante de conceptos de física. Estos resultados conﬁrman lo sugerido por diversos autores: que los conceptos no sólo deben ser deﬁnidos por su estructura, sino que se requiere considerar las situaciones en las cuales los conceptos son usados y los sistemas de representación simbólica que los estudiantes usan para pensar y escribir acerca de un concepto. Es decir, la comprensión de signiﬁcados cientíﬁcos está regulada por la interacción entre la información contenida en las situaciones y la estructura conceptual de cada estudiante (Vergnaud, 1981; 1998). Las implicaciones para futuros estudios de conocimiento previo, desde la perspectiva de la teoría de campos conceptuales, apuntan a la consolidación de una metodología para mejorar la caracterización del conocimiento previo y las representaciones que los estudiantes tienen de sus conocimientos en física (Vergnaud, 1981; 1998). Por lo tanto, apuntan a la identiﬁcación de disposiciones conceptuales para un aprendizaje signiﬁcativo progresivo de la física en el nivel universitario (Caballero, 2004; Moreira, 2000). Los resultados obtenidos con la realización de este estudio permiten una aportación preliminar de conocimiento 212 AGRADECIMIENTOS Los autores agradecen a Cecilia Fuentes, Carlos Abarzúa y Mª Angélica Osorio por sus aportaciones en la elaboración del instrumento. 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Estructura de la materia enseñada, historia de las ciencias y desarrollo conceptual del alumno, en Coll, C. (ed.). Psicología genética y educación. Barcelona: Oikos-tau. VERGNAUD, G. (1990). La théorie des champs conceptuels. Recherches en Didatique des Mathématiques, 10(23), pp. 133-170. VERGNAUD, G. (1996). Algunas ideas fundamentales de Piaget en torno a la didáctica. Perspectivas, 26(10), pp. 195-207. VERGNAUD, G. (1998). A comprehensive theory of representation for mathematics education. Journal of Mathematical Behaviour, 17(2), pp. 167-181. MOREIRA, M.A. (2002). A Teoría dos campos conceituais de Vergnaud, o ensino de ciências e a pesquisa nesta área. Investigaçoes em Ensino de Ciências, 7 (1), pp. 1-24. VYGOTSKY, L.S. (1989). El desarrollo de los procesos psicológicos superiores. Barcelona: Editorial Crítica. MOREIRA, M.A. (2004). Cambio conceptual: análisis crítico y propuestas a la luz de la teoría del aprendizaje signiﬁcati- VYGOTSKY, L.S. (1995). Pensamiento y lenguaje. Barcelona: Paidós. [Artículo recibido en abril de 2005 y aceptado en octubre de 2006] ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) 213 INVESTIGACIÓN DIDÁCTICA ANEXO 1 Ejemplo de algunas situaciones y lecturas del instrumento. Uso de conceptos en situaciones de contexto cotidiano Situación 2: Escribe dos frases en que aparezca la palabra fuerza y dos frases en que aparezca la palabra energía. 3. Para fuerza: Frase Nº 1: ___________________________________________________________ Frase Nº 2: ___________________________________________________________ 4. Para energía: Frase Nº 1: ___________________________________________________________ Frase Nº 2: ___________________________________________________________ Situación 3: Cuando yo era pequeña creía que el locutor que hablaba en la radio era un pequeño pulgarcito que llegaba a través del cable con que se enchufaba la radio. Pronto tuve que desechar mi teoría cuando mi padre llegó a casa con una ﬂamante radio a pilas. Hoy día es tan natural la presencia de equipos portátiles, incluida la telefonía móvil, que ya no nos asombramos y peligrosamente dejamos de preguntarnos, como por ejemplo: ¿Cómo se transmite la información? 5. Explica brevemente cómo es posible que la información sea trasmitida en forma inalámbrica. Explicación: ________________________________________________________________ Uso de conceptos en la comprensión de un texto Lectura Nº 2: Lee atentamente este texto tomado del libro Seis piezas fáciles de Richard Feynman, Premio Nóbel de física 1965. «Imaginemos a un niño, quizás Daniel el Travieso, que tiene unos bloques que son absolutamente indestructibles y no pueden dividirse en piezas, cada uno de ellos es igual que los otros. Daniel tiene 28 bloques y supongamos que su madre le ha dejado por la mañana con sus 28 bloques en una habitación. Al caer la tarde, sintiendo curiosidad, ella cuenta los bloques con mucho cuidado y descubre una ley empírica: haga lo que Daniel haga con los bloques ¡siempre sigue habiendo 28 bloques! Esto continúa durante varios días, hasta que un día sólo hay 27 bloques; pero tras una pequeña búsqueda la madre encuentra que hay un bloque bajo la alfombra; ella debe mirar en todas partes para estar segura de que el número de bloques no ha variado. Sin embargo, un día el número de bloques parece haber cambiado; hay sólo 26. Una investigación cuidadosa pone de maniﬁesto que la ventana estaba abierta, y al buscar fuera aparecen los otros dos bloques. Otro día, un recuento cuidadoso indica que ¡hay 30 bloques! Esto provoca una consternación considerable, hasta que la madre cae en la cuenta de que Juan vino de visita trayendo sus propios bloques, y dejó algunos en casa de Daniel. Una vez que ella se ha deshecho de los bloques extra, cierra la ventana, no deja que entre Juan, y entonces todo sigue correcto... hasta que en cierto momento cuenta y encuentra sólo 25 bloques. Sin embargo, en la habitación hay una caja, la madre quiere abrirla pero el niño grita: «No, no abras mi caja de juguetes», es decir, la madre tiene prohibido abrir la caja de juguetes. Como la madre es extraordinariamente curiosa y algo ingeniosa, ¡ella inventa una treta! Sabe que cada bloque tiene una masa de 100 gramos, así que mide la masa de la caja en un instante en que ella ve los 28 bloques en la habitación. La masa de la caja es de 600 gramos, y en cada nueva ocasión en que quiere hacer una comprobación, mide de nuevo la masa de la caja, resta 600 gramos divide por 100 y descubre lo siguiente: (Número de bloques vistos) + (masa de la caja - 600 gramos) = constante 100 gramos »En otras ocasiones parece que hay nuevas desviaciones, pero un cuidadoso estudio indica que el nivel del agua sucia de la bañera está cambiando. El niño está arrojando bloques al agua y la madre no puede verlos porque el agua está muy sucia, pero puede descubrir cuántos bloques hay en el agua añadiendo otro término a su fórmula. Puesto que la altura original del agua era 15 centímetros y cada bloque eleva el agua 0,5 centímetros, esta nueva fórmula sería: (Número de bloques vistos) + (masa de la caja - 600 gramos) (altura del agua - 15 centímetros) + = constante 0,5 centímetros 100 gramos »A medida que aumenta la complejidad de su mundo, la madre encuentra toda una serie de términos que representan formas de calcular cuántos bloques hay en los lugares donde ella no puede mirar. Como resultado, encuentra una fórmula compleja, una magnitud que debe ser calculada, que siempre tiene el mismo valor.» 214 ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) INVESTIGACIÓN DIDÁCTICA 9. ¿Qué analogías encuentras entre esta historia y la conservación de la energía? Señala y explica al menos una de ellas. Explicación: ________________________________________________ 10. ¿Cuánto vale la constante? ________________________ 11. Si en una ocasión la masa de la caja es 1.000 gramos, entonces, ¿cuál es el número de bloques vistos y el número de bloques en la caja? Nº bloques vistos: _____________ ; Nº bloques en la caja: _______________ Uso de conceptos en situaciones y problemas de contexto académico Situación 5: Si dejamos caer al mismo tiempo desde la ventana de un octavo piso dos esferas metálicas de igual tamaño, pero de distinto peso, por ejemplo, una de aluminio y otra de hierro. 13. Según tu opinión: a) La esfera de aluminio llegará primero al suelo. b) La esfera de hierro llegará primero al suelo. c) Las dos esferas llegarán al suelo al mismo tiempo. Explica lo más detalladamente posible tu respuesta, considerando cómo actúa la fuerza de gravedad en la caída de ambos cuerpos (la esfera de aluminio y la esfera de hierro). Explicación: ______________________________________________ 14. Si en el caso anterior las esferas cayeran sobre la carrocería de un auto, ¿cuál de las dos esferas abollaría más la carrocería? Explica lo más detalladamente posible tu respuesta. Explicación: ________________________________________________ Situación 10: Sacamos del congelador dos cubitos de hielo idénticos. Ponemos un cubito sobre un plato de madera y el otro sobre un plato de cobre. 18. ¿Tardarán los cubitos de hielo el mismo tiempo en descongelarse? Sí ______ ; No ______ Explica ¿Por qué? Explicación: __________________________ ENSEÑANZA DE LAS CIENCIAS, 2007, 25(2) 215 INVESTIGACIÓN DIDÁCTICA ANEXO 2 Ejemplo de criterios de corrección del desempeño de uso de conceptos en situaciones y problemas. PUNTAJE DESEMPEÑO CONCEPTOS Fuerza y energía Ondas electromagnéticas Conservación de la energía Conservación de la energía Fuerza de gravedad, masa, energía y movimiento Transferencia del calor 216 PREGUNTA 1 2 P4 No escribe frases con los conceptos de fuerza y energía con signiﬁcados en desacuerdo con signiﬁcados cientíﬁcos. Escribe frases usando el concepto energía, pero evidencia una comprensión parcial de sus signiﬁcados, que confunde con signiﬁcados de fuerza, u otros conceptos. P5 Utiliza en forma confusa No elabora un modelo conceptos asociados a ondulatorio que ondas electromagnéticas describa y explique sin lograr construir un la comunicación modelo para describir y inalámbrica. explicar la comunicación inalámbrica. P9 No identiﬁca el tema del texto ni su información para realizar analogías entre su contenido y el principio de conservación de la energía. Identiﬁca el tema del texto, pero no logra organizar la información para establecer analogías con el principio de conservación de la energía. P10 Evidencia incomprensión de signiﬁcados y aplicación de un principio de conservación en la resolución de un problema. Evidencia incomprensión de signiﬁcados y aplicación de un principio de conservación en la resolución de un problema. Evidencia conocer algunos procedimientos de cálculo que aplica mecánicamente. P14 No diferencia los efectos sobre el cuerpo (auto) golpeado y no aplica el concepto de energía para explicar estas diferencias. Identiﬁca las diferencias de efecto sobre el auto golpeado y no aplica el concepto de energía para explicar la diferencia. P18 No evidencia representaciones cientíﬁcas para el calor, ni elabora explicaciones para su transferencia. Representa el calor como energía sin asociarlo al movimiento de los átomos y moléculas de los materiales. No elabora explicaciones para su transferencia. 3 Escribe una frase con uso correcto del concepto de energía, la otra contiene signiﬁcados incorrectos que confunde con signiﬁcados de fuerza, u otros conceptos.	30,226
hal-03663553-LemaireG.-et%20al-N3AF-2022.txt_1	French-Science-Pile	Open Science	Various open science	2,022	Notes Académiques de l'Académie d'agriculture de France Academic Notes from the French Academy of Agriculture (N3AF) Synthèse. 2022. ⟨hal-03663553⟩	None	French	Spoken	6,535	9,981	HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Synthèse Nécessité d’un changement de paradigme pour étudier la nutrition et la fertilisation azotée des cultures Gilles Lemaire1, Alain Gojon2, François Gastal3, Jean-François Briat2 1. 12 rue Virecourt, 79500, Melle 2. Institut des sciences des plantes de Montpellier, Université Montpellier, CNRS, Inrae, Institut Agro, Montpellier 3. UE FERLUS, Inrae, Lusignan Correspondance : lemaire.gilles.inra@gmail.com Résumé La fertilisation azotée des cultures a longtemps été fondée sur une analyse statistique de la réponse de leur rendement à des doses croissantes de fertilisants. Ce pronostic a conduit à une pratique d’assurance où l’on applique des doses d’azote excessives, afin de ne pas pénaliser les rendements. Une méthode prévisionnelle du bilan d’azote d’une culture a été mise au point pour corriger cette tendance, mais elle reste très imparfaite. Le diagnostic de l’état de nutrition azotée d’une culture a récemment émergé, résorbant l’incertitude liée à la variabilité des conditions locales. Ce diagnostic, intégré dans des algorithmes d’aide à la décision, permet d’effectuer les apports de fertilisants seulement « là » et « quand » l’état nutritionnel des cultures le nécessite. Cette précision doit permettre d’atteindre les rendements fixés en minimisant les impacts environnementaux. Abstract Nitrogen fertilization of crops has long been based on a statistical analysis of the response of their yield to increasing doses of fertilizers. This prognosis has led to an insurance practice applying excessive doses of nitrogen so as not to penalize yields. A method for predicting the nitrogen balance of a crop has been developed to correct this trend but remains too highly imprecise. Diagnosis of the nitrogen nutrition status of a crop has recently emerged, reducing the uncertainty linked to the variability of local conditions. This diagnosis, integrated into decision support algorithms, makes it possible to apply fertilizers only “where” and “when” the nutritional status of the crops requires it. This precision must make it possible to achieve the targeted yields while minimizing the environmental impacts. Synthèse Mots clés Fertilisation azotée, bilan d’azote (méthode), courbe de réponse, courbes de dilution de N, teneur critique en N, indice de nutrition N, diagnostic N d’une culture. Keywords Nitrogen fertilization, N balance sheet method, response curves, N dilution curves, critical N concentration, nitrogen nutrition index, crop N diagnosis. Introduction La fertilisation des cultures à partir de la fabrication industrielle des engrais azotés, par le procédé Haber-Bosch, au début du 20 e siècle, a permis une extraordinaire progression de la production agricole. Un doublement de la production mondiale de denrées agricoles a été obtenu grâce à une augmentation d’un facteur 7 de l’emploi des engrais azotés (N) (Tilman et al., 2002). Cela révèle globalement une faible efficacité d’utilisation de cette ressource (Cassman et al., 2002) pourtant très coûteuse en énergie. Du fait de cette faible efficacité, cette augmentation importante de la production agricole est associée à des impacts négatifs sur les milieux terrestres et marins, liés à une trop grande circulation des formes réactives résiduelles d’azote dans les écosystèmes (Stulen, 1998 ; Galloway et Cowling, 2002 ; Beman et al., 2005 ; London, 2005). Ravier et al. (2016) ont ainsi mis en évidence l’inadéquation entre les outils d’aide à la décision mis à la disposition des agriculteurs par la recherche agronomique pour la gestion de la fertilisation azotée de leurs cultures et la maîtrise des risques environnementaux qui y sont associés. Cette note a comme objectifs (1) de rappeler les paradigmes qui ont fondé les principes sur lesquels la fertilisation azotée des cultures a Figure 1. Représentation de la variabilité de la réponse du rendement d’une culture à des apports croissants d’engrais azotés. Le rendement Y dépend de manière asymptotique de la dose d’apport Nf. En réalité, la plante réagit à une disponibilité totale en N (Nt) qui est la somme de l’apport d’engrais (Nf) et la fourniture de N par le sol (Ns). Ns étant largement inconnu, Nt l’est également, ce qui entraîne automatiquement une variation importante et inconnue du rendement Y0 qui correspond à l’absence d’apport d’engrais. De surcroît, le rendement potentiel Ymax de la culture n’étant guère prévisible, sa variation engendre une grande incertitude dans la demande en N de la culture. été historiquement bâtie, (2) de montrer en quoi ils se sont révélés défaillants pour établir des pronostics permettant de guider les agriculteurs dans leurs décisions, (3) d’analyser les connaissances acquises sur les processus d’auto-régulation de la nutrition azotée des plantes et des peuplements végétaux au niveau physiologique et moléculaire, et, enfin, (4) de montrer que de nouveaux paradigmes fondés sur la connaissance de ces processus élémentaires permettent de proposer des méthodes de diagnostic in situ de l’état de nutrition azotée d’une culture et d’en inférer des méthodes de gestion de la fertilisation azotée des cultures en temps 1. De Boussingault et von Liebig à la « méthode du bilan », 150 ans de recherche sur la nutrition et la fertilisation azotée des cultures 1.1 L’empirisme des courbes de réponse aux apports d’engrais Grâce aux travaux pionniers de Boussingault (1855) et de von Liebig (1855), la nutrition azotée des plantes et des cultures a pu être étudiée sur la base de la réponse du rendement d’une culture à des apports croissants d’engrais azotés sous forme minérale, ammoniacale ou nitrique. Il s’agissait alors d’enrichir le milieu racinaire en N minéral, afin d’engendrer une réponse de la plante en termes d’absorption, pour satisfaire une demande en N pour l’élaboration d’un rendement potentiel Ymax, luimême déterminé par le génotype et les facteurs non modifiables du milieu, le climat local (rayonnement, température, bilan hydrique, etc.) et certaines contraintes locales du sol. En effet, les travaux de physiologie végétale avaient clairement montré que la vitesse d’absorption du nitrate ou de l’ammonium par la plante variait directement comme la concentration en ces éléments dans la solution du sol (Coïc, 1956). Une approche linéaire de type « apport d’engrais » - « enrichissement du sol » « réponse de la plante » s’est donc imposée rapidement aux agronomes. Le caractère « fugace » des formes minérales d’azote dans le sol, résultant des fournitures par la minéralisation des matières organiques et des pertes par lixiviation et par volatilisation, en plus du prélèvement par les plantes, a rapidement été pris en compte, de manière à faire coïncider au mieux la date des épandages d’engrais avec la période des besoins les plus importants des cultures pour des raisons d’efficacité et aussi d’environnement (Coïc, 1974). Le paradigme de la fertilisation N des cultures a donc été fondé sur l’analyse des courbes de réponse du rendement des cultures à des apports croissants d’engrais, grâce aux outils statistiques liés à l’expérimentation au champ, afin d’en déduire un pronostic sur la dose optimale d’engrais à apporter, sachant que deux inconnues majeures devaient être estimées a priori : (1) la quantité de N minéral qui serait fournie par le sol durant la période de culture ; et (2) la demande en N de la culture elle-même déterminée par le niveau de rendement qu’elle pourrait atteindre (Figure 1). Face à la grande variabilité des courbes de réponse et à la quasi-impossibilité de pronostiquer, à partir d’elles, la dose d’engrais requise pour un objectif de rendement donné, les recommandations de fertilisation N auprès des agriculteurs ont très souvent été excessives, en vue de contrebalancer un risque de sousfertilisation. Pour les céréales à paille cela a été d’autant plus net que la crainte d’un excès d’apport de N provoquant une verse a disparu, du fait des variétés à pailles courtes et de l’utilisation des raccourcisseurs (Boiffin et al., 1982). En dehors de causes plus générales, liées aux changements d’occupation des sols, impliquant le retournement des prairies permanentes pour être remplacées par des cultures intensives de maïs, ce sont donc des excès de « fertilisation d’assurance », souvent minimes, mais répétés chaque année, qui ont contribué aux pollutions des nappes phréatiques observées depuis des années. Un excès de 20 kg N/ha sur une culture de blé ne représente qu’un surplus de moins de 10 % de la quantité de N nécessaire pour une production de 80 quintaux par hectare de grains. Cependant, si l’on admet que cet excès est entièrement lixivié par la lame d’eau drainante, le calcul aboutit à une teneur en nitrate de celle-ci dépassant le seuil de potabilité de 50 mg NO3-/L. Ce simple calcul montre donc le niveau de de la qualité des eaux impliquait donc une estimation plus fiable des doses d’engrais recommandées. 1.2 Incertitudes de pronostic de la méthode du bilan Face à cette nécessité du contrôle de la pollution des eaux, dont l’urgence est apparue dès les années 1980 (Hénin, 1992), les agronomes ont essayé de minimiser les incertitudes liées aux courbes de réponse. Suite aux travaux de Hébert (1969) et de Rémy et Hébert (1977), une approche prévisionnelle du bilan des apports, des pertes et des besoins en N d’une culture de blé a été mise en œuvre. Grâce à cette approche de type « comptable » on espérait pouvoir réduire suffisamment les incertitudes pesant sur les termes correspondant à la fourniture de N par le sol (Ns dans la Figure 1), sachant que les incertitudes pesant sur les besoins en N de la culture, liées à l’incertitude sur Ymax, pouvaient être réduites grâce à un réglage des facteurs limitants autres que N (structure du sol, limitation nutritionnelle P, K etc., maladies et ravageurs), par une meilleure conduite de la culture. Cette méthode dite du « bilan prévisionnel » permettait donc de calculer une dose optimale de N à apporter à une culture donnée comme résultat de la différence entre (1) une demande en N de la culture fondée sur un besoin, par exemple pour le blé de 3 kg N/100 kg de grain (Machet et al., 2017) pour un rendement objectif Ymax pronostiqué comme accessible sur une parcelle donnée, pour l’année en cours ; (2) une fourniture de N minéral par le sol estimée à partir d’une mesure de reliquat de N minéral présent dans le profil de sol en fin d’hiver, auquel est ajoutée une estimation de la quantité de N qui sera minéralisé dans le sol durant le reste de la période de culture ; (3) des pertes de N minéral pouvant avoir lieu lors des périodes de lixiviation, et des pertes gazeuses par volatilisation et dénitrification. Les prévisions pour chacun de ces postes du bilan ont donné lieu à de nombreux travaux des agronomes pour paramétrer des modèles de simulation en fonction de nombreuses autres variables du milieu (sol, climat), mais aussi de facteurs liés à la conduite des cultures (variétés, dates de semis, précédents culturaux, fumures antérieures, etc.), qui sont autant de sources de variations pour chacun des postes du bilan. Cet ensemble important de travaux a donné lieu à la formulation de logiciels et d’outils d’aide à la décision qui a été récemment synthétisée par Machet et al. (2017) et qui constitue une somme de références inédite sur la problématique de l’azote dans les agro-écosystèmes. Dès 1981, une analyse critique de la mise en œuvre de cette méthode avait été réalisée dans la région de la Champagne crayeuse (Boiffin et al., 1981 ; Meynard et al., 1981 ; Boiffin et al., 1982), mettant en exergue la difficulté à établir des pronostics suffisamment précis dans un certain nombre de conditions, mais ne remettant pas en cause le paradigme de base, qui est celui du bien-fondé de l’établissement d’un bilan. Les recherches en agronomie ont donc consisté à « affiner » les pronostics en faisant appel à des modèles de prévision de plus en plus paramétrés (Meynard et al., 1997). Les travaux récents de Ravier et al. (2016 ; 2018) montrent que, malgré les indéniables progrès réalisés dans les pronostics pour l’estimation des différents postes, la méthode du bilan peine toujours à réduire l’incertitude de la prévision de la dose optimale en deçà d’une certaine marge d’erreur, qui reste encore trop importante eu égard aux risques environnementaux (Meynard et al., 2002). De surcroît, l’exportation de cette approche de pronostics hors des conditions où elle a été élaborée et paramétrée se révèle difficile Synt hèse Figure 2. Dynamique de prélèvement de N par des peuplements de graminées prairiales recevant un apport de N non limitant lors d’une repousse de printemps : (a) pour une prairie de fétuque élevée (cv Clarine) pour différentes années ; (c) pour différents cultivars de fétuque élevée (cv M77 et cv Ludelle) et de dactyle (cv Cambria et cv Lucifer) pour une même année ; (b) et (c) l’expression du prélèvement de N en fonction de la dynamique de croissance (MS en t / ha ) permet d’établir une relation d’allométrie unique entre l’azote prélevé et la croissance en biomasse accumulée du peuplement végétal, quelle que soit l’origine de la variation de croissance : l’année ou le génotype (Lemaire et Salette, 1984a ; 1984b ). (Bélanger et al., 1998 ; 2001 ; Bramley et al., 2013) même quand elle est couplée avec des mesures in situ d’activité microbienne dans les sols, censées améliorer la prévision du poste « minéralisation de N par le sol » (Franzluebers et al., 2016). La difficulté à prédire les termes du bilan de N d’une culture vient essentiellement du fait que les problèmes à résoudre ne sont pas ceux d’une analyse de stocks de N, mais bien des flux de transformation de N d’une forme (organique) à l’autre (minérale), entre différents compartiment du système sol-plantemicrobiome. Les imprécisions sur les variations de stocks sont trop grandes pour en déduire ces flux. Ce sont donc les fondements même de l’approche qu’il convient de remettre en cause. 1.3 La mise en défaut d’une approche de type bilan La mise en œuvre d’une approche de « bilan de N minéral » impliquait un raisonnement de la disponibilité de N minéral dans le sol pour la plante en termes de « stock » et un cadre temporel de début et de fin de culture Notes Synthèse pour évaluer ces variations de stocks : « ouverture » et « fermeture » du bilan. Cela pouvait convenir pour les cultures annuelles comme les céréales. Dans le cadre des couverts végétaux pérennes comme les prairies, ce type d’approche est vite apparu non pertinent, pour au moins deux raisons : (1) l’absorption de N par les plantes peut se réaliser rapidement, alors qu’il n’y a pratiquement pas de stock de N minéral mesurable dans le sol, ce qui montre que la nutrition N de la plante se réalise en « flux tendu » et ne peut donc s’appréhender par une simple variation de stock ; (2) établir un bilan pour une végétation en croissance continue tout au long de l’année n’a aucun sens. Face à cette impossibilité, Lemaire et Salette (1984a ; 1984b) ont alors essayé d’expliquer la variabilité des prélèvements de N d’une prairie par les capacités de croissance du peuplement végétal, que cellesci soient déterminées par le climat de l’année ou par le génotype (espèce-cultivar) pour une année donnée. Comme cela est montré Figure 2, les différences interannuelles ou inter-génotypiques de dynamique de prélèvement de N par la végétation, lorsqu’un apport non limitant d’engrais N est effectué, sont totalement ées par la dynamique d’accumulation de biomasse par le couvert végétal. Une courbe unique de type allométrique permet de relier la dynamique de prélèvement de N et la dynamique de croissance en matière sèche (MS, exprimé en tonne de matière sèche par ha) : (1) N =a f MS 1−b Quand on fait varier la disponibilité de N par des apports d’azote, on obtient des dynamiques différentes de prélèvement de N en fonction de l’accumulation de biomasse (valeurs du coefficient af) (Figure 3). Ainsi le coefficient af de l’Équation 1 apparaît déterminé par le niveau de disponibilité de N pour la plante, lui-même dépendant du niveau de l’apport en fertilisation f. L’expression différentielle de l’équation 1 permet alors d’exprimer la vitesse de prélèvement de Figure 3. Relation dynamique entre le prélèvement d’azote (N, en kg/ha) et l’accumulation de biomasse (ici W plutôt que MS, en t/ha) d’une culture de maïs soumise à différents niveaux d’apports d’azote. Chaque niveau d’azote (f) donne lieu à une dynami que N=af (MS)1-b différente. A chaque date , il est alors possible de compare r la réponse de la culture à la quantité de N prélevée en termes de biomasse (Plénet et Lemaire, 1999). l’azote par un peuplement végétal en fonction de deux facteurs : (1) la disponibilité de N dans le sol qui détermine af, et (2) la vitesse de croissance des plantes dMS/dt : dN dN d MS =( ).( ) dt d MS dt d MS =(1−b) af MS−b (2) dt Cela a deux conséquences essentielles qui invalident toute approche de type bilan : (1) Si la vitesse de prélèvement de N par la plante, dN/dt, est liée à sa vitesse de croissance, dMS/dt, cela implique que, dans un sol donné, avec une disponibilité donnée de N minéral (valeur de af), une culture ayant une capacité de croissance plus importante qu’une autre, quelle qu’en soit la cause, le génotype ou des conditions climatiques favorables, aura non seulement une demande en N plus importante, mais également une capacité d’absorption N Synthèse accrue. Cela signifie qu’il n’y a pas d’indépendance entre « offre » et « demande » en N, car le système d’acquisition de N de la plante est en grande partie auto-régulé. En conséquence, une approche purement comptable de type « bilan » ne peut en aucun cas rendre compte de cette capacité des plantes à auto-ajuster leur capacité d’absorption de N à leur capacité de croissance. Le pronostic de la méthode du bilan prévisionnel pour évaluer le terme de la fourniture de N par le sol, Ns dans la Figure 1, uniquement à partir de paramètres du « sol », n’est donc pas pertinent, puisque ce terme N s dépend aussi de la vitesse de croissance de la plante. Ainsi certaines contraintes physiques du sol, influant sur le développement racinaire des plantes, déterminent en retour l’accessibilité de l’azote du sol pour la plante. Il devient donc difficile d’estimer Ns uniquement par les propriétés intrinsèques du sol. (2) La demande de la plante correspondant à la réalisation de sa vitesse de croissance potentielle est donc déterminée par la courbe 1−b critique d’équation N c =a c MS, telle que représentée sur la Figure 3. On voit que la demande marginale dN/dMS s’amortit au fur et à mesure que la biomasse de la culture augmente (facteur MS-b). Il s’ensuit qu’une culture avec un potentiel de croissance plus élevé qu’une autre, quelles qu’en soient les raisons (génotype ou conditions climatiques), aura une augmentation moins que proportionnelle de sa demande en N. Il n’est donc pas légitime d’utiliser une constante fixe de 3 kg / 100 kg de grain pour estimer la demande en N de la culture qu’il conviendrait de satisfaire. Ainsi on voit que l’application d’un bilan prévisionnel, ignorant la co-régulation de l’absorption de N par la croissance de la plante, engendre d’une part une sousestimation du poste « fourniture de N par le sol » (Ns dans la Figure 1) et une surestimation de la demande en N de la plante. Cela conduit à une surestimation des doses optimales d’engrais recommandées, qui sont d’autant plus importantes que les rendements potentiels (Ymax sur la Figure 1) sont élevés. La sur-fertilisation générale observée par Ravier et al. (2016) n’est donc pas due seulement à des « prises d’assurance » excessives des agriculteurs qui maximiseraient leurs rendements objectifs afin d’éviter le risque d’une sousfertilisation, mais aussi, de manière plus fondamentale, à la non prise en compte des mécanismes d’auto-régulation de l’absorption de l’azote par les plantes par une méthode purement additive comme celle du bilan prévisionnel. Devienne-Barret et al. (2000) ont utilisé le formalisme de l’Équation (2) en le couplant à celui de l’équation de Michaelis-Menten, qui relie la concentration en N du milieu à la vitesse d’absorption N de la plante. Ces auteurs ont ainsi montré qu’en intégrant ce couplage entre la vitesse de croissance de la plante et la vitesse de prélèvement d’azote, on pouvait bien rendre compte de la dynamique de nutrition azotée, pour des cultures de blé, dans une large gamme de conditions. Un certain nombre de résultats agronomiques confirment cette interdépendance entre l’offre du sol et la demande en N de la plante. Dès les années 1980, l’utilisation du marquage isotopique 15N des engrais azotés avait montré aux agronomes que les coefficients réels d’utilisation des engrais (CRU) étaient toujours inférieurs aux coefficients apparents (CAU) (Machet et al., 1987), ce qui indiquait déjà clairement qu’il n’y avait pas additivité entre les apports de N par l’engrais et la fourniture de N par le sol (Jenkinson et al., 1985). La revue récente de Recous et al. (2019) explique bien que, en réalité, la fourniture de N par le sol ne peut pas s’assimiler à un stock de N disponible pour la plante du fait du renouvellement constant minéralisation-immobilisation par la bio- Synthèse masse microbienne du sol (concept de N turnover, en anglais). Ainsi une croissance plus rapide de la plante la rend plus apte à intercepter et détourner à son profit les flux de N minéral résultant de ce renouvellement. Il s’agit maintenant d’analyser par quels mécanismes physiologiques la plante est capable de modifier sa capacité d’absorption de N minéral du sol en fonction de sa propre demande. 2. Les interactions plantes-microorganismes du sol déterminent la disponibilité de l’azote du sol pour les racines Les organismes vivants du sol influencent la disponibilité minérale pour les racines des plantes. Ces communautés d’organismes, telles que le microbiote (Deveau et Martin, 2016) et la micro- et macro-faune, établissent de multiples compétitions trophiques entre elles. En conséquence, la composition de ces communautés est modifiée, entraînant la modification de la disponibilité des nutriments pour les plantes. Ces événements se produisent dans une zone particulière du sol, la rhizosphère, qui est formée et influencée par les racines et les micro-organismes associés. Ces équilibres déterminent la disponibilité des éléments minéraux de la rhizosphère, qui, dans une boucle rétroactive, font évoluer les équilibres trophiques des sols (Briat et al., 2020) (Figure 4). Une partie notable du carbone fixé par les feuilles, par la photosynthèse, est secrétée par les racines dans la rhizosphère sous forme de molécules organiques. Ces exsudats racinaires représentent entre 5 à 30 % des produits de la photosynthèse, soit entre 1 et 3 tonnes de C par hectare et par an. Par exemple, on estime qu’un hectare de maïs produisant 18 tonnes de matière sèche aérienne allouera 7 tonnes de C aux racines, dont 2,3 tonnes seront excrétées dans la rhizosphère (Waligora, 2014). Figure 4. Le carbone du gaz carbonique (CO2) de l’atmosphère est incorporé dans des molécules organiques (saccharides) via la photosynthèse qui se déroule dans les feuilles des plantes. Une partie de ce carbone est transloquée vers les racines et sécrétée (exsudats) dans l’interface sol-racine (rhizosphère ). Ce carbone organique sert de nutriment aux microbiotes du sol, dont les communautés participent à la minéralisation de l’azote organique contenu dans la matière organique du sol (MO) ; cette dernière, en retour, peut être absorbée par les plantes. Ce cycle C/N est fondé sur des équilibres dynamiques qui reposent sur des interactions/ compétitions trophiques entre les communautés des organismes du sol entre elles et avec les racines des plantes. À cette interaction directe entre les plantes et le microbiome s’ajoute la fourniture indirecte d’azote minéral aux plantes due à la « boucle microbienne » qui résulte de la consommation des organismes du microbiome par des protozoaires et des nématodes (Briat et al., 2020). Synthèse La grande variabilité de ces chiffres s’explique par le fait que de nombreux éléments liés au sol, à l’environnement, et aussi à la nature même de la plante, modulent les différentes proportions de C allouées aux parties aériennes, aux racines, et exsudées dans la rhizosphère. Les plantes pérennes, par exemple, investissent beaucoup plus dans leur système racinaire que les plantes annuelles et y consacrent donc plus de C. L’environnement joue également un rôle déterminant. En cas de stress hydrique, de ion ou de carence nutritionnelle, les plantes dépensent plus d’énergie dans leurs racines et y allouent donc plus de C (Waligora, 2014). Ces exsudats nourrissent une importante communauté microbienne à l’interface solracine : le microbiote racinaire. Celui-ci contient des micro-organismes bénéfiques, qui aident les végétaux dans leur acquisition de l’eau et des minéraux. Notamment les bactéries du microbiote et les champignons des mycorhizes sont impliqués dans des processus de minéralisation de N organique (contenu dans la matière organique du sol) en N minéral (ammonium et nitrate), qui sera in fine absorbé par les plantes (Figure 4). Les plantes ont donc la capacité de changer leur environnement racinaire et, par conséquent, de modifier leur microbiome (Briat et Lemaire, 2021). Indépendamment des interactions trophiques directes entre plantes et organismes du sol, des interactions indirectes existent. Par exemple, des protozoaires et des nématodes se nourrissent des bactéries et du mycélium des mycorhizes, et rejettent ainsi du N minéral dans le sol, absorbable par les racines (Trap et al., 2016). Ce mécanisme connu sous le nom de « boucle microbienne » aboutit à la modification de la composition des communautés microbiennes et des flux de nutriments minéraux du sol. Ces interactions entre les protozoaires, les mycorhizes arbusculaires et les racines conduisent à une augmentation de la biomasse et à l’accumulation d’azote dans les plantes (Plassard et al., 2015 ; Briat et al., 2020) (Figure 4). Les plantes sont donc en compétition avec les micro-organismes du sol pour utiliser le N minéral qu'ils produisent à partir de la matière organique, mais en synergie avec eux en leur fournissant du C par les exsudats racinaires dérivant de la photosynthèse. Les plantes pilotent ainsi à leur profit, au moins en partie, le renouvellement minéralisationréorganisation de l’azote. 3. La co-régulation de l’absorption de N par l’offre dans le sol et par la croissance de la plante La capacité des plantes à ajuster leur prélèvement racinaire de N aux besoins nutritionnels induits par la croissance est attestée par deux types d’observations : (1) à régime d’alimentation constant en N, les variations de croissance induisent des variations similaires de la vitesse d’absorption de nitrate (Imsande et Touraine, 1994), et (2) des plantes soumises à des régimes d’alimentation en N très différents sont capables de maintenir une même vitesse d’absorption de nitrate. Clément et al. (1978) ont spectaculairement établi ce dernier point en montrant qu’une diminution d’un facteur 1 000 de la concentration en nitrate dans la solution nutritive fournie à des plantes de ray-grass (et renouvelée en permanence) n’entraînait à terme qu’une diminution de moins de 20 % de l’absorption totale de N et de la croissance. Cela illustre clairement que la prise en compte de la disponibilité externe en N (l’offre du sol) ne permet pas, à elle seule, de prédire quel sera le prélèvement de ce N par les plantes. Même si l’activité instantanée des systèmes d’absorption de N dépend fortement de sa concentration externe (suivant une cinétique comparable à celle des enzymes, telle que décrite par le formalisme de Michaelis- ( hèse Menten), les plantes modulent l’efficacité de ces systèmes pour compenser les variations de concentration externe et maintenir la vitesse d’absorption au niveau requis pour alimenter la croissance. Les processus qui permettent cette modulation commencent à être bien connus, y compris au niveau le plus fin des voies moléculaires de signalisation (Nacry et al., 2013 ; Vidal et al., 2020). De manière schématique, ils correspondent à trois types de mécanismes de régulation, qui agissent aussi bien au niveau physiologique (abondance et activité des systèmes protéiques d’absorption racinaire de N) qu’au niveau développemental (taille et architecture du système racinaire). Le premier mécanisme de régulation correspond à un contrôle systémique exercé par des signaux spécifiques du statut nutritionnel en N de la plante entière. Ce contrôle vise à stimuler l’absorption de N lorsque la plante est en situation de carence en N, ou au contraire à la freiner en situation de satiété. Le second mécanisme correspond aussi à un contrôle systémique, mais cette fois-ci par des signaux induits par l’activité photosynthétique des parties aériennes. Ce contrôle assure la coordination entre fixation de C par les parties aériennes et prélèvement de N par les racines. Enfin le troisième mécanisme est celui de perception du nitrate lui-même, qui permet aux racines de déterminer où et à quelle concentration il est présent dans le sol. Les deux premiers mécanismes assurent le pilotage de l’absorption de N par la croissance alors que le troisième assure ce pilotage en fonction de l’offre du sol. C’est parce que les plantes intègrent en permanence la mise en œuvre de ces trois types de mécanismes qu’a émergé le concept de co-régulation de l’absorption de N par l’offre du sol et la croissance de la plante. Le cadre conceptuel décrit ci-dessus est générique dans la mesure où les trois types de régulation ont été observés chez toutes les espèces étudiées. Néanmoins, c’est chez la Figure 5. Mécanismes moléculaires assurant la régulation de l’absorption racinaire de nitrate chez Arabidopsis thaliana. La disponibilité externe du nitrate est perçue par le transporteur-senseur NRT1.1. La signalisation feuilles-racines de la demande en N est assurée par les polypeptides mobiles CEPD et CEPDL. La signalisation feuilles-racines de l’activité photosynthétique met en jeu le polypeptide mobile HY5. Les polypeptides NRT1.1, CEPD, CEPDL et HY5 sont des régulateurs positifs de NRT2.1, qui est une composante majeure du système d’absorption racinaire du nitrate (Fileur et al., 2001). plante modèle Arabidopsis thaliana que la connaissance des mécanismes physiologiques et moléculaires impliqués est la plus avancée. En guise d’illustration, les paragraphes suivants et la Figure 5 résument l’état de cette connaissance pour le contrôle du transporteur racinaire de nitrate NRT2.1, qui constitue une composante majeure du système protéique d’absorption de nitrate chez Arabidopsis (Filleur et al., 2001). Le contrôle systémique de NRT2.1 par le statut N de la plante entière fait appel à un mécanisme de signalisation à longue distance (racines-feuilles et feuilles-racines), médiée par des peptides et des Synthèse polypeptides induits par la carence en N au niveau des différents organes (Ohkubo et al., 2017). Parmi ces polypeptides, trois glutarédoxines (CEPD1, CEPD2 et CEPDL) constituent les signaux systémiques transportés des feuilles vers les racines pour réguler l’expression et l’activité de NRT2.1 (Ota et al., 2020). Ce module de régulation polypeptidique intègre donc le statut nutritionnel des différents organes, et assure la transmission de ces informations aux racines pour activer NRT2.1 et ainsi compenser une carence en N subie par la plante. Le contrôle systémique de l’absorption racinaire de nitrate par la photosynthèse implique vraisemblablement une composante énergétique, associée à la métabolisation, dans les racines, des sucres produits par les parties aériennes. Cependant il est maintenant acquis qu’il implique également une ou plusieurs composantes de signalisation, activées par la lumière et les sucres (Sakuraba et Yanagisawa, 2018). Dans ce cas, également, le mécanisme de signalisation met en jeu le transport des feuilles vers les racines d’un polypeptidesignal, qui correspond au facteur de transcription HY5. La synthèse de HY5 dans les feuilles est stimulée par la lumière, et le signal agit dans les racines en induisant l’expression de gènes de la nutrition minérale, notamment NRT2.1 (Chen et al., 2016). En parallèle, la métabolisation des sucres dans les racines active une voie de signal au niveau du cycle des pentoses-phosphates, dont le détail moléculaire est encore inconnu, mais qui aboutit également à l’induction de NRT2.1 (Lejay et al., 2008). Ces mécanismes font que l’expression et l’activité de NRT2.1 dans les racines suivent fidèlement les variations de la photosynthèse, et permettent d’expliquer que toute augmentation de la fixation de C dans les parties aériennes entraîne, dans les heures qui suivent, une augmentation équivalente du prélèvement de nitrate par les racines (Gastal et Saugier, 1989). Le troisième contrôle, lié à la perception du nitrate en tant que molécule-signal, est de nature différente des deux premiers, car il n’a pas de composante nécessairement systémique et agit surtout localement. Cette perception du nitrate, notamment celui qui est présent dans le milieu extérieur, est assurée par des capteurs spécifiques, dont le mieux connu est NRT1.1 (Bouguyon et al., 2015). NRT1.1 a en fait une double activité : il est un transporteur racinaire assurant une partie de l’absorption de nitrate, mais il joue également un rôle crucial de régulation, en activant des voies de signalisation dans les cellules racinaires, en réaction à la présence du nitrate à l’extérieur de ces cellules. Or l’activation locale de ces voies de signalisation est indispensable à l’action des deux autres mécanismes de régulation. Ainsi la stimulation de NRT2.1 par les signalisations systémiques N et C ne se produit que dans des racines au contact du nitrate externe. Il s’agit d’un mécanisme important d’adaptation à l’hétérogénéité spatiale de disponibilité du nitrate dans le sol, qui assure que la stimulation des capacités d’absorption racinaire de nitrate en réponse à une carence en N ou à une augmentation de la photosynthèse ne se fera que là où le nitrate est effectivement présent dans le sol. Bien que surtout détaillés chez Arabidopsis thaliana, les mécanismes de signalisation décrits précédemment existent chez les autres espèces. Par exemple, le module de régulation impliquant les CEPD/CEPDL a aussi été décrit chez les légumineuses (Taleski et al., 2018), et la signalisation nitrate dépendante de NRT1.1 est active chez le riz Oryza sativa, où NRT1.1 est un déterminant majeur de l’efficience de l’utilisation de l’azote au champ (Wang et al., 2020). Ainsi une avancée majeure des dix dernières années a été la découverte que la co- Synthèse régulation de l’absorption racinaire de N par la croissance de la plante et l’offre du sol repose sur l’action de protéines de signalisation, qui véhiculent (parfois à longue distance) les informations sur le statut N de la plante, son activité photosynthétique (les déterminants de la croissance) et la richesse du milieu en nitrate (l’offre du sol). C’est l’intégration de ces informations qui permet à la plante de prélever, là où il est le plus abondant, le N dont elle a besoin pour entretenir sa croissance. 4. Les courbes de dilution de N : un outil de diagnostic pour piloter la nutrition N des cultures Les mécanismes moléculaires de régulation de l’absorption d’azote par les plantes tels que décrits précédemment permettent de considérer l’équation (1) comme la résultante de l’auto-ajustement du prélèvement d’azote d’une culture en fonction de sa propre demande et de la disponibilité de N dans le sol. L’équation (1) peut être transformée pour exprimer la diminution de la teneur en N de la plante (%N) au cours de sa croissance : %N = af MS−b (3) 10 Le coefficient 1/10 permet d’exprimer %N en g N/100 g de biomasse alors que MS est exprimée en t/ha. Dans ce cas, af /10 représente la teneur en N de la plante lorsque la biomasse de la culture est de 1 t/ha. Le coefficient -b représente la diminution de la teneur en N de la plante lorsque la biomasse augmente : c’est le phénomène de dilution. Comme le montre la Figure 6, il est alors possible de déterminer la teneur en N critique de la plante correspondant à la teneur en N minimum (%N) nécessaire pour obtenir la biomasse maximum et de définir ainsi une courbe de dilution critique pour une espèce Figure 6. Détermination de la teneur en N critique d’une culture au cours de sa croissance. La valeur de %Nc est déterminée par l’intersection (points noirs) entre la droite simulant la réponse W = f(%N) et la verticale représentant la valeur maximale de W (Wmax) lorsque l’azote n’est plus limitant. La diminution de %Nc avec l’augmentation de W permet alors de déterminer la courbe de dilution critique de N pour le maïs (Plénet et Lemaire, 1999). : %N c ac MS−b (4) De nombreux travaux ont permis de déterminer cette courbe critique pour un grand nombre d’espèces : graminées fourragères (Lemaire et Salette, 1984 a ; 1984 b), luzerne ( Lemaire et al., 1986), blé Triticum aestivum (Justes et al., 1994), maïs Zea mais L. (Plénet et Lemaire, 2000), colza Brassica napus (Colnene et al., 1998), tournesol Helianthus annuus (Debaecke, 2012), entre autres. Une méthode d’analyse bayesienne des données expérimentales a été mise au point réc emment pour déterminer l’incertitude concernant l’ estimation des paramètres ac et b, permettant ainsi de faire des analyses des Notes Académiques , 1, génotype-milieu de plus large ampleur (Makowski et al., 2020 ; Ciampitti et al., 2021). Lemaire et Gastal (1997), puis Lemaire et al. (2008) ont créé un corpus théorique justifiant le choix d’une formulation allométrique de la courbe de dilution critique. Il apparaît que deux processus complémentaires sont à la base de cette dilution de N : (1) Une relation d’allométrie entre le compartiment métabolique MSm de la plante (tissus impliqués dans la croissance : parenchymes photosynthétiques et tissus méristématiques) ayant une teneur en N (%Nm) élevée, et le compartiment structural (MS)s (tissus de soutien et vaisseaux conduct eurs impliqués dans l’architecture de la plante) ayant une teneur faible en N (%Ns) conduisant à une diminution du rapport MSm / MSs lorsque la plante augmente de taille et donc de masse [MS = MSm+MSs] selon le modèle de Caloin et Yu (1984). (2) Une att énu ation de la teneur en N des feuilles (%Nm) au sein du couvert végétal en relation avec le profil d’extinction de la lumière au fur et à mesure du développement du couvert végétal et, donc, de l’augmentation de biomasse de la culture conformément aux travaux de Hirose et Werger (1987), de Charles-Edward et al. (1987) et de Lemaire et al. (1991). Cet effet s’ajoute à un effet ontogénétique lié à l’âge des feuilles, et est réversible si l’on remet les feuilles à la lumière. Ainsi la courbe de dilution critique, telle qu’exprimée dans l’équation (4), est la traduction des propriétés émergentes du couvert végétal. Celles-ci résultent des mécanismes physiologiques sous-jacents. Ceux-ci s’exercent au niveau moléculaire (voir paragraphe précédent) pour ajuster la nutrition azotée à la nutrition carbonée de chaque plante et, donc, à sa propre capacité de croissance. Cette courbe traduit donc à la fois les mécanismes de régulation de l’absorption de N par les plantes, aussi bien que les mécanismes de répartition au sein de Figure 7. Détermination de l’indice de nutrition N d’une culture (INN) comme le rapport entre la teneur en N de la plante (%Nact) dans une culture ayant une biomasse donnée (MSact) et la teneur critique en N (%Nc) correspondant à cette même biomasse telle que calculée par la courbe de dilution N critique (INN = %Nact / %Nc). Ainsi une valeur de INN > 1 indique une nutrition N excessive tandis qu’une valeur INN < 1 indique une déficience en N (Lemaire et al., 2008). la plante et entre les plantes au sein d ’un même couvert végétal en relation avec la ré partition de la lumière .	37,945
https://openalex.org/W2760654863	OpenAlex	Open Science	CC-By	2,017	Regular Exposure to Cowbells Affects the Behavioral Reactivity to a Noise Stimulus in Dairy Cows	Julia Johns	English	Spoken	9,376	16,524	R Julia Johns1,2, Sophie Masneuf 2,3, Antonia Patt 2,4 and Edna Hillmann2,5* The 85-dB stimulus increased arousal and avoidance compared with the 65-dB stimulus, with bell experience and earplugs leading to a general decrease in avoidance of the stimulus. This may reflect an altered acoustic perception of the playback stimulus in dairy cows that are routinely exposed to bells. In alpine regions, cows are often equipped with bells during pasture season to ensure that farmers can locate them. Constant exposure to the chime of a bell may affect cows’ acoustic perception in general. The aim of this study is to test whether routine bell exposure affects the reactivity to a noise stimulus and might be associated with hearing impairment in cows. For the assessment, behavioral and cardiac indicators were used as indirect measures of hearing capacity. Cows that were either used to wearing a bell or not were exposed to a playback of low and high amplitude (=varying loudness). In addition, we tested whether wearing earplugs, mimicking hearing impairment, reduced the cows’ reactivity toward the playback. On 24 farms, half of them routinely using cowbells, 96 Brown Swiss cows were tested in a 2 × 2 factorial cross-over design (65 or 85 dB, without or with earplugs) in a balanced order. The effects of bell experience, amplitude, and earplugs on the latency to the first behavioral and cardiac response to a 5-s playback were analyzed using linear mixed-effects models, considering depen- dencies within the data set. Cows reacted faster without earplugs and when they were exposed to 85 dB compared with 65 dB. The proportion of cows leaving the feeding rack after onset of the playback was reduced by bell experience and earplugs and was increased when exposed to 85 dB compared with 65 dB. Exposure without earplugs to 85 dB but not to 65 dB increased heart rate. Heart rate and heart rate variability indicated increased sympathetic activation during the exposure to 85 dB compared with 65 dB. In general, behavioral and cardiac indicators did not indicate severe hearing impairment due to routine bell exposure. The 85-dB stimulus increased arousal and avoidance compared with the 65-dB stimulus, with bell experience and earplugs leading to a general decrease in avoidance of the stimulus. This may reflect an altered acoustic perception of the playback stimulus in dairy cows that are routinely exposed to bells. Original Research published: 29 September 2017 doi: 10.3389/fvets.2017.00153 Original Research Keywords: avoidance, behavior, cattle, earplug, noise, playback Edited by: Laura Ann Boyle, Teagasc, The Irish Agriculture and Food Development Authority, Ireland Reviewed by: Courtney L. Daigle, Texas A&M University, United States Renate Luise Doerfler, Technische Universität München, Germany Correspondence: Edna Hillmann edna.hillmann@hu-berlin.de Correspondence: Edna Hillmann edna.hillmann@hu-berlin.de Specialty section: This article was submitted to Animal Behavior and Welfare, a section of the journal Frontiers in Veterinary Science Received: 04 May 2017 Accepted: 05 September 2017 Published: 29 September 2017 R Julia Johns1,2, Sophie Masneuf 2,3, Antonia Patt 2,4 and Edna Hillmann2,5* Julia Johns1,2, Sophie Masneuf 2,3, Antonia Patt 2,4 and Edna Hillmann2,5* 1 Faculty of Organic Agricultural Sciences, Farm Animal Behavior and Husbandry Section, University of Kassel, Witzenhausen, Germany, 2 Ethology and Animal Welfare Unit, Department of Environmental Systems Science, Institute of Agricultural Sciences, ETH Zurich, Zurich, Switzerland, 3 Department of Neurology, University Hospital Zurich, Zurich, Switzerland, 4 Institute of Animal Welfare and Animal Husbandry, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Celle, Germany, 5 Animal Husbandry, Albrecht Daniel Thaer-Institute of Agricultural and Horticultural Sciences, Humboldt-Universität zu Berlin, Berlin, Germany In alpine regions, cows are often equipped with bells during pasture season to ensure that farmers can locate them. Constant exposure to the chime of a bell may affect cows’ acoustic perception in general. The aim of this study is to test whether routine bell exposure affects the reactivity to a noise stimulus and might be associated with hearing impairment in cows. For the assessment, behavioral and cardiac indicators were used as indirect measures of hearing capacity. Cows that were either used to wearing a bell or not were exposed to a playback of low and high amplitude (=varying loudness). In addition, we tested whether wearing earplugs, mimicking hearing impairment, reduced the cows’ reactivity toward the playback. On 24 farms, half of them routinely using cowbells, 96 Brown Swiss cows were tested in a 2 × 2 factorial cross-over design (65 or 85 dB, without or with earplugs) in a balanced order. The effects of bell experience, amplitude, and earplugs on the latency to the first behavioral and cardiac response to a 5-s playback were analyzed using linear mixed-effects models, considering depen- dencies within the data set. Cows reacted faster without earplugs and when they were exposed to 85 dB compared with 65 dB. The proportion of cows leaving the feeding rack after onset of the playback was reduced by bell experience and earplugs and was increased when exposed to 85 dB compared with 65 dB. Exposure without earplugs to 85 dB but not to 65 dB increased heart rate. Heart rate and heart rate variability indicated increased sympathetic activation during the exposure to 85 dB compared with 65 dB. In general, behavioral and cardiac indicators did not indicate severe hearing impairment due to routine bell exposure. INTRODUCTION Bells seem to be a relevant noise factor for cows considering that cows are exposed routinely and for a longer period of time to the chime of bells (1). The aim of this study is to test whether routine bell exposure affects the reactivity to a noise stimulus and be associated with hearing impairment in cows. Behavioral and cardiac indicators were used as indirect measures for the assessment of cows’ hearing capacity. Thus, we examined the reactivity toward noise of low (65 dB) and high (85 dB) ampli- tude in bell-experienced and bell-inexperienced cows on 24 Swiss dairy farms. We additionally tested whether mimicking hearing impairment using earplugs would reduce the reactivity to the sounds. We hypothesized that cows that had been exposed regularly to a bell on alpine pastures (bell-experienced cows) would show reduced reactivity toward these sounds (increased latency of the first behavioral reaction, reduced avoidance and heart rate, and increased heart rate variability) contrarily with cows that were only equipped with a bell as heifers or never before (bell-inexperienced cows). Contrary, we expected that cows would show increased reactivity in response to a stimulus of high amplitude (decreased latency of the first behavioral reac- tion, increased avoidance and heart rate, and reduced heart rate variability) compared to a stimulus of low amplitude. Further, we expected that cows without earplugs would also show increased reactivity toward these sounds (decreased latency of the first behavioral reaction, increased avoidance and heart rate, and reduced heart rate variability) compared to cows with earplugs. Altogether, if earplugs do not diminish the reaction of a given cow, this might be an indicator of either a low-reactive animal, well habituated to noise or hearing impairment. Noise-induced hearing loss is one of the most common causes of exogenously acquired sensorineural hearing loss in adult humans (6). Although anatomic differences among mammal species lead to differences in hearing capacities (7), the basic physiologic processes underlying the detection and sensation of sound are essentially identical between humans, dogs, cattle, and mice (8–11). Considering that cows can hear sounds between 23 Hz and 35 kHz, with the highest sensitivity at 8 kHz, and are able to detect sounds at −11 dB, i.e., amplitudes the human ear cannot detect (11), the continuous exposure to bells during pasturing season might impair the cows’ hearing capacity. INTRODUCTION Johns J, Masneuf S, Patt A and Hillmann E (2017) Regular Exposure to Cowbells Affects the Behavioral Reactivity to a Noise Stimulus in Dairy Cows. Front. Vet. Sci. 4:153. doi: 10.3389/fvets.2017.00153 In alpine regions, cows are often equipped with a bell throughout the summer season to ensure that farmers can locate their animals on the wide alpine pastures, many areas that are obstructed from view. The chime of these cowbells is characterized by high and varying amplitudes from 90 to 113 dB at a distance of 20 cm, the approximated distance between the bell and the cows’ ears (1). Goats September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 1 Bell-Experience Affects Reaction to Noise Johns et al. to such hazardous noise can result in irreversible hearing loss and that even a single intense sound event can cause hearing loss and tinnitus (32, 33). have been found to show higher behavioral arousal when being exposed to the playback of a bell compared to the playback of a uniform sinusoidal sound, indicating that the bell sound might be more aversive to goats than the uniform sound. With repeated exposure, goats habituated to both stimuli (2). A widespread solution used to protect humans and also horses from noise exposure by using hearing protection devices such as earplugs (34, 35). Commercially available earplugs for horses are made of memory foam (35). In cattle, acoustic ear- phones were inserted into the ear canal and were held in position with either silicone earplugs or earplugs of compressible foam while measuring brainstem auditory evoked potentials (BAEPs) (36–38). Such earplugs occlude the ear canal and attenuate background noise. So far, little research has been conducted investigating the effect of noise on the hearing capacities of animals. Kenneled dogs that were constantly exposed to noise between 100 and 108 dB for 6 months developed hearing loss as indicated by measurements of the auditory brainstem response (ABR) (3). In mice, ABR recordings showed that a single exposure to noise of 100 dB for 2 h induced temporary hearing loss (4), and an exposure to noise of 110 dB for 60 min even induced permanent hearing loss (5). g Although some studies on the general hearing capacity of cows are available, to our knowledge, no studies exist on hearing capacities of cows that have been exposed to noise in general. INTRODUCTION Behavioral indicators such as the acoustic startle response (12–15) or avoidance reactions (16, 17) have been used as an indi- rect but non-invasive test of hearing capacity in earlier studies. The acoustic startle response is an electromyographic response, which in rodents is elicited by stimuli with an amplitude of more than 80 dB (18, 19), and the latency is very short [5–10 ms for the electromyographically measured response in different muscles (20–22)]. Behaviorally, a startle response is defined as a cross-species response to an intense and abrupt stimulus (23) and as any first reaction of any part of the body, such as body movements, movements of limbs and facial movements, or any first behavioral reaction to sound stimulation (24). Therefore, the latency to the first behavioral reaction, e.g., sudden head move- ments in response to an acoustic stimulus can be used as a proxy for the induction of a startle response. In addition, avoidance reactions in response to an acoustic stimulus, e.g., increasing the distance between the source and oneself, indicate that the stimulus is perceived as aversive by the animals (25–27). In addition, cardiac parameters such as heart rate and heart rate variability can be used to assess arousal induced by noise (2, 28). If cardiac parameters indicate an arousal due to noise exposure, it can be assumed that the animal perceives the noise as aversive (23). Noise exposure is often accompanied by an increase of heart rate in humans (29, 30). Lee et al. (31) evaluated instant responses of the autonomic nervous system to short- duration noises using heart rate variability analysis. The results indicated that, compared with background noise of 38 dB, exposure to noise between 50 and 80 dB increased sympathetic activity as indicated by a higher ratio of low-frequency (LF) to high-frequency (HF) spectral power. For humans, “hazardous” noise is defined as sounds that exceed 85 dB over a typical 8-h workday (32, 33). It has been shown that constant exposure ANIMALS, MATERIALS, AND METHODS Ethical approval to conduct the study was obtained from the Zurich Cantonal Veterinary Office, Switzerland (approval No. 77/2012). Frontiers in Veterinary Science \| www.frontiersin.org Acoustic Stimulus experimental cows were selected randomly. On 12 farms, cows were used to wearing a bell either every year for 4–5 months during the summer season or all year round (48 cows on 12 farms, “experienced”). On the other 12 farms, cows had no or very little experience with wearing a bell, having either never worn a bell before or only once for 4–5 months when they were 1 year old (48 cows on 12 farms, “inexperienced”). On all 24 farms, cows were kept in cubicle housing systems with a feeding rack and headlocks. All cows were familiar with being locked in the feeding rack during feeding. They were fed with hay, fresh grass or a mixed ration of hay, corn, and grass silage. Feed and water were provided ad libitum. The cows were milked twice a day. Each cow was exposed to a pink noise stimulus broadcast four times for 5 s in a 2 × 2 factorial design: each cow was tested with and without earplugs and at 65 (A-weighting scale, A) and 85 dB (A) (Figure 2; each amplitude with two phases: without and with earplugs), i.e., each of the bell-experienced and bell- inexperienced cows were exposed to each amplitude (65 and 85 dB) twice, once with and once without earplugs (4 trials per cow, see Experimental Procedure). The A-weighting scale assigns low weights to the LF tones, to which the human ear and the ears of some animals are less sensitive, and high weights to the HF tones, to which humans are more sensitive (39). Pink noise is characterized by uniformly distributed energy throughout the range of human hearing, approximately 20 Hz–20 kHz. Most people perceive pink noise as having uniform spectral power density, i.e., the same loudness at all frequencies (40). The noise stimulus was automatically generated by a Tone Generator Pro v1.0.8 (Performance Audio®) for iPhone®. We chose the pink noise for three reasons: first, it has no biologic relevance for the cows, and therefore, the reactions to the acoustic stimuli were mostly likely limited to the perception of the acoustic stimuli per se. Second, it contains all frequencies from 20 Hz to 20 kHz at the same amplitude and should thus match the cows’ (potential) hearing capacity. Third, it was a novel acoustic stimulus for all cows, and none of them was ever exposed to it before. Acoustic Stimulus The volume of the playback was set at a level that ensured an amplitude of either 65 or 85 dB (A) at the feeding rack at approximately 60 cm above the floor (i.e., the estimated position of the cows’ heads when feeding). The amplitude was measured with a precision noise level measuring instrument with integrated long-term storage (SoundTest-Master, Laserliner®, Umarex GmbH & Co. KG, Arnsberg, Germany). The background noises at the farms measured before the start of the experiments ranged from 40 to 60 dB (A). Animals, Housing, and Managementh , g, g The study was performed between September and November 2013 on 24 Swiss farms, with 96 multiparous Brown Swiss cows that were between 3 and 10 years of age. The owners of the cows gave permission to conduct the study on their farms. The size of the farms varied between 8 and 100 animals. On each farm, four September 2017 \| Volume 4 \| Article 153 2 Johns et al. Bell-Experience Affects Reaction to Noise Test Area within Barnh The experiment was carried out at the feeding rack of each farm during the course of 1 day. A 5-m section of the feeding rack served as test area for the experiment. During the experiment, only the four experimental cows had access to this area, one cow at a time. To record the animals’ behavior, a video camera with an integrated microphone (Canon® Legria FS 200 digital camera) was mounted on a tripod and positioned in front of the separated feeding rack area (Figure 1). The acoustic stimulus was transmitted via two loudspeakers (Edifier® S2000v, Edifier International, Hong Kong, China) that were positioned on the floor next to the camera. The loudspeakers, the tripod, and cables were hidden behind a visual cover of white fabric (Figure 1) and placed on a board (approximately 1.5 m × 1.5 m). The front edge of the board with the loudspeakers was placed in the feed- ing alley at a distance of 2.15 m from the feeding rack to ensure that the intended amplitude reached the cows’ ears. Figure 1 \| Technical equipment. Technical equipment behind a visual cover of white fabric. C, camera; S, speakers; X, experimenters were hidden behind the visual cover during playback. Experimental Procedure In the morning around 8 O’clock, the visual cover of white fabric with all technical equipment was set up. Before the start of the experiment, the four experimental cows were habituated to the technical equipment (Figure 1), the thorax belt to measure Figure 2 \| Earplugs. Cow with earplug (70 mm × 40 mm × 40 mm) made of memory foam (left) in the ear canal (right). Figure 1 \| Technical equipment. Technical equipment behind a visual cover of white fabric. C, camera; S, speakers; X, experimenters were hidden behind the visual cover during playback. Figure 2 \| Earplugs. Cow with earplug (70 mm × 40 mm × 40 mm) made of memory foam (left) in the ear canal (right). September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 3 Bell-Experience Affects Reaction to Noise Johns et al. order (Figure 3). To avoid exposure to the acoustic stimuli prior to testing, the other experimental cows were either separated in an outdoor paddock (n = 11 farms) or fitted with earplugs and led to the farthest part of the stable (n = 13 farms). heartbeat parameters (see Heartbeat Measurements) and the earplugs (Figure 2). Each experimental cow was exposed to the pink noise stimulus in four trials (Figure 3). The order in which the cows were tested was chosen randomly before the start of the experiment. To reduce handling of the cows, the two playbacks during which cows were equipped with earplugs were tested in consecutive trials. Thus, each cow was equipped with the earplugs only once during the experiment with the order of phases and amplitudes chosen randomly for each animal. p A handful of concentrate was sprinkled on the usual feed during trials to enhance feeding motivation of the experimental cow. As soon as the experimental cow was feeding calmly for at least 1 min, the headlocks were carefully opened by the second experimenter, and the playback was switched on for 5 s (20 ± 3 s after opening the headlocks). Start and stop of the playback were controlled manually by the experimenter using a mobile phone (iPhone 4 s®, Apple Inc.). After the end of the playback, behavioral observations and heartbeat measurements continued for 2 more minutes (playback + 2 min = trial). Each experimental cow was tested in all four trials in one session. Behavior Behavioral analyses were conducted by two different people (person 1 analyzed the latency to the first reaction, and person 2 analyzed avoidance) who had not participated in the conduct of the experiments. They did not know the farms and were not aware of the aim of the study. However, as they needed to record the behaviors related to the start of the playback, they were aware that acoustic stimuli were involved but were blind to the bell experience of the animals, the amplitude, or if the animals were equipped with earplugs. In addition, to test intraobserver reliability for the assessment of latency to the first reaction, person 1 assessed 96 trials twice. Experimental Procedure When the session was finished, the thorax belt (and earplugs, depending on phase order) was removed, and the cow was allowed to rejoin the herd. If a cow did not start feeding within 1 min after offering the con- centrate, no playback was conducted, and another experimental cow was tested. On each farm, the four experimental cows were led to the sepa- rated 5-m section of the feeding rack, restrained in the headlocks (for feeding), and fitted with a thorax belt. Then, they were habitu- ated to the earplugs. The earplugs (70 mm × 40 mm × 40 mm; Figure 2) were made of memory foam (polyurethane with addi- tional non-toxic chemicals increasing its viscosity and density; Vibraplast AG®, Aadorf, Switzerland). Earplugs were compressed and placed into the ear canal during feeding, where they expanded and plugged the ear canal. To avoid experimental cows being irritated by a sudden impairment of hearing, the first earplug was positioned into the left ear for 15 min before the second earplug was placed into the right ear. Experimental cows were left undisturbed for 30 min with both thorax belt and earplugs in place. During these 30 min, the two earplugs had to remain in position for at least 15 min. If the animal shook its head resulting in the loss of an earplug, the earplug was repositioned. During the first 15 min, the first earplug had to be repositioned 1.3 times (minimum 1 and maximum 6 times), and during the following 30 min, an earplug had to be repositioned 1.9 times (minimum 1 and maximum 7 times). At the end of the habituation phase, all experimental cows accepted the earplugs and continued to feed calmly. After the habituation phase, the earplugs were removed, and the experimental cows were allowed to rejoin the herd for 1 h. To ensure that experimental cows were motivated to feed, cows had no access to feed while rejoining the herd.t Heartbeat Measurements Heartbeat measurements were recorded using Polar® Equine (Polar® Elektro Europe BV, Zug, Switzerland), allowing a non- invasive measurement of heartbeats (41, 42). To increase the electrode–skin contact, electrode gel (Anandic Medical Systems AG/SA, Feuerthalen, Switzerland) was used. A thorax belt with two integrated electrodes was fixed around the torso directly behind the forelegs. One electrode was positioned ventrally on the left side of the sternum and the other one at a given dis- tance by the thorax belt on the left thoracic wall. A receiver for recording the data was placed between the two electrodes. The thorax belt was additionally protected by an elastic belt of about 5 cm width. The heartbeat was recorded for 1 min of continuous feeding before the playback started, during the playback (5 s), and 2 min after the playback. Data were downloaded onto a computer via a base station using Bluetooth (Polar® Team2 Pro, version 1.3.0.3). Statistical analyses were performed in R 3.3.1 and 3.3.2 (R Core Team 2016, 43). We used the agreement package (51) to check the intraobserver agreement concerning the latency to the first reaction. To adequately reflect dependencies in the experimental design (nesting, repeated measurements), linear mixed-effects models were used to evaluate the latency to the first reaction and heartbeat measurements with the lmer meth- ods from “lme4” and “lmerTest” packages, respectively (52). The occurrence of leaving the feeding rack was analyzed using a generalized linear mixed-effects model [glmer method from package “lme4” (53)]. Here, we used odds ratios [exponential function of the regression coefficient associated with a one- unit increase in the exposure (54)] to additionally quantify the effect sizes.if Analysis of cardiac data was carried out using the programs Polar® ProTrainer 5 Equine Edition (version 5.35.161, © Polar Electro Europe AG, Zug, Switzerland) and R 3.2.3 (43). Root mean square of successive differences (RMSSD) of heartbeats reflects changes in the vagosympathetic balance that are vagally mediated (42) and represents parasympathetic activity, whereas standard deviation of heartbeats (SDNN) is a more complex parameter reflecting vagal and sympathetic activation (42, 44). The spectral measures HF band and LF band are highly correlated with the time domain-related measures RMSSD and SDNN, respectively (45, 46). Consequently, the ratio between RMSSD and SDNN can be used as an indicator of changes of the vagosympathetic balance in the organism (47–49), similar to the ratio of HF and LF (44). Latency to the First Reactionhi After the 1-h break, experimental cows were individually led into the separated area of the feeding rack, restrained in the headlocks, and fitted with the earplugs depending on stimulus The latency to the first reaction was described as time (seconds) it took the experimental cows from the onset of the playback Figure 3 \| Experimental procedure. The two playbacks (65 and 85 dB) with earplugs were tested in consecutive trials; the order of amplitude within a phase (without or with earplugs) and between phases was chosen randomly for each cow. Figure 3 \| Experimental procedure. The two playbacks (65 and 85 dB) with earplugs were tested in consecutive trials; the order of amplitude within a phase (without or with earplugs) and between phases was chosen randomly for each cow. September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 4 Bell-Experience Affects Reaction to Noise Johns et al. (Polar® ProTrainer 5 Equine Edition, version 5.35.161). Data with an error rate of more than 5% were excluded from the analysis according to the studies by Hagen et al. and Gygax et al. (46, 50). In addition, data of one cow were excluded from analysis due to extremely high heart rate regardless of experimental treatment. This led to the exclusion of 286 trials and a remaining sample size of 98 trials. to react with a clearly visible change of the ear posture, paus- ing (cow stopped her current behavior and froze shortly), or a sudden head movement (cow stopped feeding and raised its head quickly). The latency to the first reaction was analyzed in slow motion (0.5× real time) from video using VLC media player® (version 2.2.4 Weatherwax, VideoLAN, Paris, France) and a stopwatch for Smartphone Shift5.1® (Android version 5.1, SHIFT GmbH, Falkenberg, Germany) for 15 s after onset of the playback. to react with a clearly visible change of the ear posture, paus- ing (cow stopped her current behavior and froze shortly), or a sudden head movement (cow stopped feeding and raised its head quickly). The latency to the first reaction was analyzed in slow motion (0.5× real time) from video using VLC media player® (version 2.2.4 Weatherwax, VideoLAN, Paris, France) and a stopwatch for Smartphone Shift5.1® (Android version 5.1, SHIFT GmbH, Falkenberg, Germany) for 15 s after onset of the playback. Data Analysish The experimental design resulted in a sample size of 384 trials (i.e., 24 farms × 4 experimental cows × 4 trials). However, five cows older than 10 years were excluded from data analysis to avoid the interference of age-dependent hearing impairment (presbycusis) with experimental treatments. One cow had to be excluded from data analysis due to technical problems and another cow due to not feeding at all. In eight trials (five cows on five farms), cows did not start feeding again after a playback exposure. In these cases, we had to quit the session. All these cows had no or very little experience with wearing a bell. Technical problems with video recording occurred in another seven trials. Thus, the total sample size was 89 cows on 24 farms in 341 trials. Latency to the First Reactionhi The number of R-R intervals [heart rate in beats per minute (bpm)], the RMSSD (ms) and the ratio between RMSSD and SDNN (RMSSD/SDNN) were calculated. The 1 min preceding the playback and the first minute of the trial were both divided into first (0–20 s), middle (21–40 s), and last (41–60 s) 20 s. The middle 20 s of the minute preceding the playback was then cho- sen as reference value for heartbeat parameters. This reference time window was compared with the first time window (0–20 s) after start of the playback (trial value) by calculating the ratios of heart rate (bpm), RMSSD (ms), and RMSSD/SDNN between reference and trial value. Avoidance An avoidance reaction to the playback was recorded whenever a cow left the feeding rack (completely withdrawing the head from the feeding rack) within 60 s after onset of the playback using INTERACT® (Mangold International GmbH, version 9.0.7, Arnstorf, Germany). A duration of 60 s (=5 s playback + 55 s) as a time window that may reflect a response to the playback and not to any other stimuli, e.g., environmental sounds was considered. Frontiers in Veterinary Science \| www.frontiersin.org September 2017 \| Volume 4 \| Article 153 Heartbeat Measurements An increased ratio between RMSSD and SDNN indicates that vagosympathetic balance is more shifted toward parasympa- thetic activation, whereas a lower ratio indicates a shift toward sympathetic activation (49). Full models consisted of the fixed-effects “bell experience” (factor with two levels: experienced, inexperienced), “earplugs” (factor with two levels: with, without), and “amplitude” (factor with two levels: 65 and 85 dB) and all possible interactions. Models were reduced in a stepwise backward procedure. P > 0.05 was used as criterion for exclusion of non-significant interactions. P values were calculated based on likelihood ratio tests. Trial nested in individual identity nested in farm served as random effects. To satisfy model assumptions, heartbeat parameters were log transformed. Automatic correction of the tachograms was carried out using the correction routines included in the Polar® software September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 5 Bell-Experience Affects Reaction to Noise Johns et al. Heartbeat Measurements In the description of the results of heartbeat measurements, a ratio >1 indicates that the trial value was greater than the refer- ence value, and vice versa for a ratio <1.h In the figures, descriptive data and model estimations are shown; in the text, model estimations and odds ratios were used to interpret the results. The mean absolute heart rate was 78.9 bpm (minimum: 54 bpm, maximum: 180 bpm) in the minute before and 79.5 bpm (minimum: 54 bpm, maximum: 180 bpm) in the minute after playback exposure and showed a large interindi- vidual variability. When cows were exposed to 85 dB without earplugs, heart rate during the first 20 s after onset of the playback was increased compared with the baseline heart rate (amplitude × earplugs: F1,100.1 = 3.99; P = 0.048; Figure 6A). We found no effect of bell experience (F1,16.8 = 0.44; P = 0.515) on heart rate response.h Latency to the First Reactionh The intraobserver agreement for the assessment of the latency to the first reaction was good, with a concordance coefficient of 0.91. the first reaction was good, with a concordance coefficient of 0.91. As expected, cows reacted faster when exposed to the 85-dB stimulus compared with the 65-dB stimulus (F1,212.1 = 56.65, P < 0.001) and slower when equipped with earplugs (F1,214.6 = 65.05, P < 0.001). Bell experience did not affect the latency to the first reaction (F1,24.5 = 0.5, P = 0.486, Figure 4). The RMSSD ratio following the playback at 85 dB was increased compared with the RMSSD ratio following the playback at 65 dB (F1,102.8 = 5.58; P = 0.020). Further, RMSSD ratio was lower when wearing earplugs than when not wearing earplugs (F1,48.4 = 6.22; P = 0.016). We found no effect of bell experience (F1,56.0 = 0.09; P = 0.769) on RMSSD response (Figure 6B).h Avoidanceh The probability for a cow to leave the feeding rack within 60 s after exposure to the sound stimulus was strongly increased when cows were exposed to 85 dB compared with 65 dB (odds ratio = 3.20; χ2 = 17.29; P < 0.001; Figure 5). Similarly, the probability to leave the feeding rack was reduced when wearing earplugs (odds ratio = 0.30; χ2 = 17.63; P < 0.001; Figure 5) and when cows were bell experienced (odds ratio = 0.33; χ2 = 4.92; P = 0.027; Figure 5). The RMSSD/SDNN ratio was reduced when cows were exposed to 85 dB compared with 65 dB (F1,107.2 = 13.45; P < 0.001), and it was slightly increased by earplugs (F1,53.2 = 3.27; P = 0.076; Figure 4 \| Latency to the first reaction 15 s after onset of the playback stimulus depending on bell-experienced (no, yes), earplugs (without, with) and amplitude (65, 85 dB). Descriptive data are presented as box plots indicating observed median, first and third quartiles, and absolute range of data. Solid lines show the model estimation and dotted lines show the lower and upper 95% confidence intervals. Figure 4 \| Latency to the first reaction 15 s after onset of the playback stimulus depending on bell-experienced (no, yes), earplugs (without, with) and amplitude (65, 85 dB). Descriptive data are presented as box plots indicating observed median, first and third quartiles, and absolute range of data. Solid lines show the model estimation and dotted lines show the lower and upper 95% confidence intervals. September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 6 Johns et al. Bell-Experience Affects Reaction to Noise Figure 5 \| Avoidance. Proportion of cows leaving the feeding rack within 60 s after playback depending on bell-experienced (no, yes), earplugs (without, with), and amplitude (65, 85 dB). Solid lines show the model estimation, and dotted lines show the lower and upper 95% confidence intervals. Figure 5 \| Avoidance. Proportion of cows leaving the feeding rack within 60 s after playback depending on bell-experienced (no, yes), earplugs (without, with), and amplitude (65, 85 dB). Solid lines show the model estimation, and dotted lines show the lower and upper 95% confidence intervals. Figure 6C). Again, no effect of bell experience was detectable (F1,60.0 = 0.07; P = 0.797). showed faster transit times, indicating an escape reaction (55). Avoidanceh Heifers exposed to playbacks of people shouting (86 dB) and metal-on-metal clanging (85 dB) moved more often compared to no noise (56). Talling et al. (57) and Geverink et al. (58) found aberrant behavior when sound levels were up to 85 dB in pigs. Further, pregnant ewes that were exposed to noises with amplitudes increasing from 45 to 95 dB reduced their feeding time and increased the time spent inactive (59). MacKenzie et al. (60) found that hens showed avoidance to high-intensity noises (90–110 dB). Consequently, in this study, a shorter latency to the first reaction and increased avoidance indicate that cows perceived the acoustic stimuli as aversive.h Frontiers in Veterinary Science \| www.frontiersin.org DISCUSSION The experimental setting in this study allowed an assessment of the cows’ reactions to acoustic stimuli as both amplitude and earplugs affected all outcome variables in a meaningful way. However, none of the cows seemed to have severe hearing impairment as earplugs diminished the reactions of the cows to the acoustic stimuli. Overall, the cows responded to the 85-dB stimulus stronger than to the 65-dB stimulus. Bell experience and earplugs reduced avoidance of the noise stimulus. The high-amplitude stimulus (85 dB) triggered a shorter latency to the first reaction and increased heart rate and avoidance than the low-amplitude stimulus (65 dB). With earplugs, the latency to the first reaction was longer, and the cows showed less avoidance of the sound stimulus. However, bell experience had no effect on the latency to the first reaction. The amplitudes of cowbells that are traditionally used on Alpine pastures vary between 90 and 113 dB (1). However, we did not measure the amplitudes of the bells that the experienced cows were wearing during pasture season. It is likely that there were differences in the amplitudes of the bells worn by the expe- rienced cows. If so, this might explain the wide variability of responses observed in the experienced cows to a certain extent. The amplitudes of cowbells that are traditionally used on Alpine pastures vary between 90 and 113 dB (1). However, we did not measure the amplitudes of the bells that the experienced cows were wearing during pasture season. It is likely that there were differences in the amplitudes of the bells worn by the expe- rienced cows. If so, this might explain the wide variability of responses observed in the experienced cows to a certain extent. Furthermore, the increased heart rate during the 85-dB stimulus without earplugs corresponds to previous studies that found noises with higher amplitudes evoking stronger cardiac reactions than noises with lower amplitudes in pigs and humans. Talling et al. (57) found that pigs had a higher heart rate when i Our results correspond to previous studies that found noises with higher amplitudes (e.g., >85 dB) evoke stronger behavioral reactions than noises with lower amplitudes in farm animals. September 2017 \| Volume 4 \| Article 153 DISCUSSION In humans, dogs and mice hearing loss has been shown to occur after they were exposed to noise with amplitudes similar to those of cowbells. Further, exposure time in these studies was similar to the time cows are exposed to bells while on pasture (3–5, 33). However, given the effect of the earplugs and the fact that all cows in our study showed a reac- tion to the playback in at least one trial, none of them seemed to have a severe hearing impairment. Figure 6 \| Heart rate, root mean square of successive difference (RMSSD), and RMSSD/standard deviation of heartbeats (SDNN). Ratio of (A) heart rate, (B) RMSSD, and (C) RMSSD/SDNN between playback and reference situation depending on bell-experienced (no, yes), earplugs (without, with), and amplitude (65, 85 dB). Descriptive data are presented as box plots indicating observed median, first and third quartiles, and absolute range of data. Solid lines show the model estimation, and dotted lines the lower and upper 95% confidence intervals. The dark red line represents the ratio value in case of identical values during trial and reference. Considering the experimental setting in retrospect, the pink noise stimulus might not have been ideal to assess the full hearing capacity of cows. Given that cows can hear sounds between 23 Hz and 35 kHz (10), a high-pass filter that blocks LFs and passes HFs (64), or a stimulus that contains only higher frequencies might be more suitable to test cows’ hearing capacity in future studies since hearing impairment is associated with reduced sensitivity to HFs. In addition, the 85-dB stimulus used in our study might have been too loud to detect subtle differences in hearing capaci- ties. On the other hand, the amplitude used to elicit an acoustic startle response was more than 80 dB in other studies (12, 18, 19, 29, 31). Thus, inferring information about hearing capacity from a latency to the first reaction to an acoustic stimulus as a proxy for the induction of a startle response might partly be misleading when using a high-amplitude stimulus, and other behavioral indicators have to be found. Furthermore, the studies mentioned above (3–5, 33) used standardized clinical hearing tests (BAEP and ABR). BAEP are bioelectric waves that can be recorded within 10 ms after an auditory stimulus and are used to assess auditory function (65). DISCUSSION When exposed to a playback of background noise recorded in milking parlors and broadcast at 85 dB in a raceway, heifers Furthermore, the increased heart rate during the 85-dB stimulus without earplugs corresponds to previous studies that found noises with higher amplitudes evoking stronger cardiac reactions than noises with lower amplitudes in pigs and humans. Talling et al. (57) found that pigs had a higher heart rate when September 2017 \| Volume 4 \| Article 153 September 2017 \| Volume 4 \| Article 153 7 Bell-Experience Affects Reaction to Noise Johns et al. Figure 6 \| Heart rate, root mean square of successive difference (RMSSD), and RMSSD/standard deviation of heartbeats (SDNN). Ratio of (A) heart rate, (B) RMSSD, and (C) RMSSD/SDNN between playback and reference situation depending on bell-experienced (no, yes), earplugs (without, with), and amplitude (65, 85 dB). Descriptive data are presented as box plots indicating observed median, first and third quartiles, and absolute range of data. Solid lines show the model estimation, and dotted lines the lower and upper 95% confidence intervals. The dark red line represents the ratio value in case of identical values during trial and reference. to a novel object elicited an increase in RMSSD, indicating an increased parasympathetic activity. In the current study, heart rate after the playback at 85 dB was increased most likely due to enhanced sympathetic activity, and RMSSD ratio was increased due to an increased parasympathetic activity. At the same time, RMSSD/SDNN ratio was reduced, indicating a stronger activa- tion of the sympathetic branch, overall. Consequently, the pink noise stimulus might have been both sudden and novel (31, 63). Cows avoided the playback stimulus less when wearing earplugs, or when their farm of origin used bells during the pasture season (bell experienced), which may reflect an altered acoustic perception of the playback stimulus due to routine bell exposure. Furthermore, only cows from farms that did not use bells for their cows did not start feeding again after being exposed to the playback. This may indicate that the playback was perceived as aversive by these cows. Although the experimental setting of our study did not allow us to conclusively assess the hearing capacity of the cows, a reduced reactivity to the acoustic stimuli due to hearing impairment of bell-experienced cows can- not be excluded completely. Frontiers in Veterinary Science \| www.frontiersin.org REFERENCES 15. Braff D, Stone C, Callaway E, Geyer M, Glick I, Bali L. Prestimulus effects on human startle reflex in normals and schizophrenics. Psychophysiology (1978) 15:339–43. doi:10.1111/j.1469-8986.1978.tb01390.x 1. Johns J, Patt A, Hillmann E. Do bells affect behaviour and heart rate variability in grazing dairy cows? PLoS One (2015) 10:e0131632. doi:10.1371/journal. pone.0131632 16. Pryor GT, Rebert CS, Howd RA. Hearing loss in rats caused by inhalation of mixed xylenes and styrene. J Appl Toxicol (1987) 7:55–61. doi:10.1002/ jat.2550070110 2. Johns J, Patt A, Hillmann E. Effects of sounds of different quality on the behaviour and heart beat parameters of goats. Appl Anim Behav Sci (2015) 165:72–80. doi:10.1016/j.applanim.2015.02.005 f 17. Knudsen FR, Enger PS, Sand O. Avoidance responses to low frequency sound in downstream migrating Atlantic salmon smolt, Salmo salar. J Fish Biol (1994) 45:227–33. doi:10.1111/j.1095-8649.1994.tb01302.x 3. Scheifele P, Martin D, Clark JG, Kemper D, Wells J. Effect of kennel noise on hearing in dogs. Am J Vet Res (2012) 73:482–9. doi:10.2460/ajvr. 73.4.482 18. Pilz PK, Schnitzler HU, Menne D. Acoustic startle threshold of the albino rat (Rattus norvegius). J Comp Psychol (1987) 101:67–72. doi:10.1037/0735-7036.101.1.67 4. Chuang S, Lei S, Xuejun J, Shiming Y, Ke L. Effects of moderate noise exposure on hearing function in C57BL/6J mice. J Otol (2014) 9:81–5. doi:10.1016/ S1672-2930(14)50019-0 19. Valsamis B, Schmid S. Habituation and prepulse inhibition of acoustic startle in rodents. J Vis Exp (2011) 55:e3446. doi:10.3791/3446 20. Cassella JV, Harty TP, Davis M. Fear conditioning, pre-pulse inhibition and drug modulation of a short latency startle response measured electromyo- graphically from neck muscles in the rat. Physiol Behav (1986) 36:1187–91. doi:10.1016/0031-9384(86)90499-3 5. Park SN, Back SA, Park KH, Seo JH, Noh HI, Akil O, et al. Comparison of functional and morphologic characteristics of mice models of noise- induced hearing loss. Auris Nasus Larynx (2013) 40:11–7. doi:10.1016/j. anl.2011.11.008 21. Pilz PK, Caesar M, Ostwald J. Comparative threshold studies of the acoustic pinna, jaw and startle reflex in the rat. Physiol Behav (1988) 43:411–5. doi:10.1016/0031-9384(88)90112-6 h 6. Miller MH, Schein JD. Hearing Disorders Handbook. San Diego, CA: Plural Publishing, Inc (2008). 275 p. 7. Fay RR. Hearing in Vertebrates: A Psychophysics Databook. Winnetka, IL: Hill-Fay Associates (1988). 22. Koch M, Schnitzler HU. The acoustic startle response in rats – circuits medi- ating evocation, inhibition and potentiation. Behav Brain Res (1997) 89:35–49. doi:10.1016/S0166-4328(97)02296-1 8. Strain GM, Myers LJ. Hearing and equilibrium. In: Reece WO, editor. FUNDING Ethical approval to conduct the study was obtained from the Zurich Cantonal Veterinary Office, Switzerland (approval No. 77/2012). The project was funded by budget funds of ETH Zurich and Felsentorstiftung. The project was funded by budget funds of ETH Zurich and Felsentorstiftung. ACKNOWLEDGMENTS We thank all farmers for their participation in this study, their openness and trust toward us, many informative discussions, and their hospitality. We also thank Simon Peter Luzi for his assistance with data collection and analysis as well as Karin Scheuss and Sarina Fetscher for analyzing the video recordings. We are grateful to PD Dr. Lorenz Gygax for his support during the statistical analyses. AUTHOR CONTRIBUTIONS to the 85-dB stimulus stronger than to the 65-dB stimulus and using cowbells with lower amplitudes might be advantageous. Study design: JJ and EH. Data collection: JJ, SM, and AP. Data analysis: JJ and EH. Manuscript drafting: JJ, AP, and EH. Critical revisions of the manuscript: SM, AP, and EH. Final approval: JJ, SM, AP, and EH. CONCLUSION Our results demonstrated that acute exposure to the 85-dB pink noise stimulus triggered increased arousal and avoidance compared with the 65-dB stimulus. Heart rate and heart rate variability indicated increased sympathetic activation during the exposure to 85 dB compared with 65 dB. Bell experience and wearing earplugs led to a generally decreased avoidance of the stimulus compared with bell-inexperienced cows and with cows not wearing earplugs. This may reflect an altered acoustic perception of the playback stimulus due to noise habituation, a low-reactive animal in general, or impaired hearing capacity when routinely exposed to bells. DISCUSSION Due to the influence of excessive muscle movements on the measurement, it is necessary for the subject to be motionless during the procedure (37). Clinically, therefore, sedation is needed when measuring BAEP in young children or animals. Accordingly, BAEP measurements need to be conducted in a veterinary hospital rather than on-farm to be able to monitor the animals more closely. Figure 6 \| Heart rate, root mean square of successive difference (RMSSD), and RMSSD/standard deviation of heartbeats (SDNN). Ratio of (A) heart rate, (B) RMSSD, and (C) RMSSD/SDNN between playback and reference situation depending on bell-experienced (no, yes), earplugs (without, with), and amplitude (65, 85 dB). Descriptive data are presented as box plots indicating observed median, first and third quartiles, and absolute range of data. Solid lines show the model estimation, and dotted lines the lower and upper 95% confidence intervals. The dark red line represents the ratio value in case of identical values during trial and reference. exposed to very loud (97 dB) compared with loud (85 dB) stimuli. In addition, a sudden loud sound (110 dB, 1–20 kHz, 0.15 s) evoked an immediate increase of heart rate in humans (29). In our study, changes in heart rate, RMSSD, and the ratio between RMSSD and SDNN were rather small (i.e., ratios varied little and were close to 1), thus the cardiac reaction following the playback has to be interpreted with caution. However, Désiré et al. (61, 62) showed that the sudden appearance of an object elicited an increase in heart rate in lambs most likely due to enhanced sym- pathetic activity, i.e., increased arousal. In addition, the exposure Altogether, we could not clinically assess the hearing capacity of bell-experienced cows. Nevertheless, our indicators showed that routine bell exposure led to a mitigation of the behavioral response to a novel acoustic stimulus. Overall, the cows responded September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 8 Bell-Experience Affects Reaction to Noise Johns et al. REFERENCES Dukes’s Physiology of Domestic Animals. Ithaca, NY: Cornell University Press (2004). p. 852–64. 23. 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Conflict of Interest Statement: The authors declare that the research was con- ducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. 46. Hagen K, Langbein J, Schmied C, Lexer D, Waiblinger S. Heart rate variability in dairy cows-influences of breed and milking system. Physiol Behav (2005) 85:195–204. doi:10.1016/j.physbeh.2005.03.019 47. Patt A, Gygax L, Wechsler B, Hillmann E, Langbein J, Keil NM. Context specificity of the ANS stress response during two regrouping experiments in goats. Front Vet Sci (2016) 3:58. doi:10.3389/fvets.2016.00058 Copyright © 2017 Johns, Masneuf, Patt and Hillmann. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. 48. Kuhne F, Hoessler JC, Struwe R. Behavioral and cardiac responses by dogs to physical human-dog contact. J Vet Behav (2014) 9:93–7. doi:10.1016/j. jveb.2014.02.006 September 2017 \| Volume 4 \| Article 153 Frontiers in Veterinary Science \| www.frontiersin.org 10	14,663
https://openalex.org/W4234206386_1	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	974	1,645	Mercados y Negocios (1665-7039) 1665-7039 impreso 2594-0163 electrónico Año 22, Núm. 44, junio-diciembre (2021) Carta del editor Mercados y Negocios ha renovado su imagen, estamos usando OJS 3 y el equipo editorial se ha centrado en los contenidos en línea, así como en la difusión de los artículos en distintos medios electrónicos. Esperamos puedan seguirnos en Facebook y en LinkedIn. También tenemos el placer de presentarles el número 44. Este número está compuesto por cuatro artículos de investigación y una reseña bibliográfica que han sido sometidos al proceso de evaluación por pares. Aprovechamos para agradecer a los árbitros por su compromiso y dedicación; así como al Consejo de Redacción que contribuye a la calidad de los contenidos de cara a un mayor impacto de los trabajos publicados en Mercados y Negocios. Estamos muy contentos de formar parte del Índice de revistas certificadas por REDALYC, Sistema de Información Científica. La tarea de transformación editorial no ha sido fácil, y en esta última etapa ha exigido modificar el flujo del proceso editorial. Esta transformación ha sido coronada con la oportunidad de, a través de Marcalyc, ofrecer al lector el contenido de los últimos años en diferentes formatos. Estos contenidos se pueden consultar en el sitio facilitado por Redalyc. https://www.redalyc.org/revista.oa?id=5718 Speculation about the Future of Capitalism, fue escrito por Kamal Fatehi, y es el primer artículo de este número El autor reflexiona sobre el futuro y lo desconocido que resulta. Aborda la discusión sobre la posibilidad de eventos futuros como un ejercicio que carece de evidencia que respalde el contenido de dicha discusión. Un argumento sin evidencia es mera conjetura y especulación. Sin embargo, en la especulación, se puede usar patrones de eventos existentes y pasados para sugerir la dirección de posibles asuntos futuros. Este artículo especula sobre los cambios futuros que puede sufrir el capitalismo. Al hacerlo, utiliza los desarrollos pasados y presentes de las ciencias y los valores cambiantes. Ofrece dos escenarios posibles. El primer escenario es optimista y el segundo no es muy optimista. Incluso se podría decir que es una predicción pesimista. El segundo artículo ha sido escrito por Claudia Leticia Preciado Ortiz y se titula Quality and Use of Mobile Applications for Transportation Service: Influence on Satisfaction. En él se presenta los resultados de una investigación que tuvo como principal objetivo analizar los factores que influyen en la satisfacción y la intención de continuar con el uso de aplicaciones Carta del editor móviles de transporte en los jóvenes universitarios de Guadalajara, Jalisco, México. El método fue cuantitativo. La investigadora utilizó 144 respuestas válidas y el modelado de ecuaciones estructurales de mínimos cuadrados parciales (PLS-SEM) para probar el modelo. El software empleado fue SmartPLS 3. Los resultados indicaron que la calidad de diseño, la calidad de la información y la calidad del sistema son predictores influyentes en la satisfacción. De acuerdo con las conclusiones de Preciado Ortiz, las empresas que ofrecen transporte individual de pasajeros mediante una app móvil han aumentado en los últimos años, generando una fuerte competencia tanto entre las marcas existentes como con los taxis tradicionales establecidos. Este segundo artículo aporta información nueva y reciente para los directores de marketing y académicos sobre el comportamiento del usuario de apps en la industria del transporte. 2 Creencias y actitudes del consumo religioso en jóvenes de Hidalgo, México, es el trabajo que ocupa el tercer espacio en Mercados y Negocios. Su autor es Omar Daniel Meneses Armenta. El objetivo del artículo es conocer qué creen y qué actitudes de consumo tienen los jóvenes frente a las organizaciones religiosas en el estado de Hidalgo, México. Meneses Armenta desarrolló una investigación descriptiva con una encuesta con escala de Likert, de intervalo y de proporción a 383 jóvenes de la Universidad Autónoma del Estado de Hidalgo. El principal resultado al que llega el autor en esta investigación es que, dependiendo de la religión, los jóvenes tienden a tener mayor dedicación hacia la misma, y que las generaciones han creado cambios. La conclusión es que, a diferencia del catolicismo, los jóvenes de las otras religiones demuestran mayor enfoque, sin embargo, el cambio generacional afecta el interés por la religión de modo decreciente, afectando también el consumo por motivos religiosos. No solo en la religión, también en otros productos o servicios, la lealtad hacia la marca es frágil. Manuela Badillo Gaona escribió Investigación en la acción: propuesta de formación alternativa, el cuarto trabajo. El objetivo de este artículo es analizar una propuesta de formación docente con perfil investigador, a fin de que incida en sus actividades de gestión, innovación y docencia. Badillo Gaona presenta un estudio cualitativo con base en la lógica, la crítica-dialéctica e investigación-acción. Los sujetos que colaboraron en la investigación fueron nueve docentes de los tres niveles educativos. La autora aplicó un cuestionario abierto, el procesamiento de datos fue heurístico-reflexivo y usó cuadros de análisis. Los resultados conducen al reconocimiento y consolidación de un Seminario Taller de Formación en y para la investigación y una propuesta de un Diplomado en la que se plasma la experiencia vivida. MERCADOS y Negocios Sánchez, J. La reseña Negociación tridimensional, es presentada por Renata Kubus. En esta se hace un análisis de la obra, mostrando la forma en que los autores logran su objetivo al transmitir al lector la importancia de considerar la negociación en tres dimensiones para alcanzar la transformación del escenario hacia situaciones que generen mayor valor para las partes involucradas. Agradecemos a nuestros autores, lectores, a los miembros del Comité Científico, del Consejo de Redacción, y a la comunidad científica internacional por continuar confiando y apoyando este proyecto que se ha materializado en el número 44. Confiamos en superar los estándares de calidad que nos hemos impuesto y a nunca quedar satisfechos con lo alcanzado. Dr. José Sánchez Gutiérrez Editor 3 Año 22, N. 44, julio-diciembre 2021: 1-4 Carta del editor 4 MERCADOS y Negocios.	14,603
https://openalex.org/W2192208731_1	Spanish-Science-Pile	Open Science	Various open science	2,015	La voz inconformista de Patrick Kavanagh	None	English	Spoken	6,105	10,600	Estudios Irlandeses, Number 10, 2015, pp. 35-43 __________________________________________________________________________________________ AEDEI La voz inconformista de Patrick Kavanagh Maximiliano Brina Magíster, Universidad de Buenos Aires, Argentina Copyright (c) 2015 by Maximiliano Brina. This text may be archived and redistributed both in electronic form and in hard copy, provided that the author and journal are properly cited and no fee is charged for access. Resumen. Entre los poetas irlandeses de la primera mitad del siglo XX, Patrick Kavanagh, poeta rural y autodidacta, destacó por su visión crítica, entre el rechazo y la desconfianza, de programas nacionalistas literarios y políticos. Frente a la representación cuasi mítica del campesinado construida por el Irish Literary Revival, luego recuperada y utilizada por los republicanos como la síntesis de lo irlandés, una suerte de “ser nacional” construido por elites urbanas, Kavanagh presenta una mirada única, que oscila entre la desolación y la sátira. La propuesta de este artículo es examinar, a partir de “The Great Hunger” (1942) y poemas previos, la forma en que Kavanagh desmonta la construcción de esa visión urbana del campesinado irlandés. Palabras clave. Patrick Kavanagh, poesía, “The Great Hunger”, nacionalismo, religión, representación, tradición, campesinado, pastoril. Abstract. Patrick Kavanagh, rural and self-taught poet, stands out among the Irish poets of the early twentieth century for his critical consideration of literary and political nationalist programs. Facing the quasi mythical representation of the peasantry built by the Irish Literary Revival, which was later used by the Republicans as a synthesis of Irishness, Kavanagh offers a unique opinion, ranging between despair and satire. The purpose of this article is to examine, from “The Great Hunger” (1942) and previous poems, the way Kavanagh disassembles this urban vision of the Irish peasantry. Key Words. Patrick Kavanagh, Poetry, “The Great Hunger”, Nationalism, Religion, Representation, Tradition, Peasantry, Pastoral. altivo, gerifalte Poeta, ayúdame: a luz más cierta, súbeme: la patria no es la tierra, el hombre no es el árbol: ayúdame a vivir sin suelo y sin raíces: móvil, móvil: sin otro alimento y sustancia que tu rica palabra: palabra sin historia, orden verbal autónomo, engañoso delirio: poema: imaginación y razón en ti se aúnan a tu propio servicio: palabra liberada de secular servidumbre Juan Goytisolo – Reivindicación del Conde Don Julián Nacido lejos de Dublín, en el condado de Monaghan donde residió hasta fines de la década de 1930, Patrick Kavanagh tuvo una formación literaria autodidacta paralela al trabajo rural. Su visión crítica del Irish Literary ___________________________________ ISSN 1699-311X Revival y la entronización de un campesinado cuasi-mítico construido desde la ciudad (“I’m the only man who has written in our time about rural Ireland from the inside”, citado en Campbell 2003: 42) así como la posición crítica, 36 entre la desconfianza y el rechazo, frente a proyectos nacionales políticos y literarios a los que opuso el parochialism y la sátira en poemas como “The Wake of the Books (A Mummery)” o “Who killed James Joyce?” hacen de este poeta un punto de quiebre dentro del panorama literario irlandés de la primera mitad del siglo XX. Mi propuesta de trabajo es concentra en un corpus conformado por poemas producidos entre 1929 y 1949, con particular interés en “The Great Hunger”, de 1942. La particularidad de Kavanagh impone la formalidad de una breve contextualización, un marco desde el cual abordaré al poeta para describir desde dónde construye su voz (y contra quién). En primer lugar, el Irish Literary Revival o simplemente Revival, movimiento que entre las últimas décadas del siglo XIX y las primeras del XX recuperó el interés por la herencia y tradición gaélica. Se destaca la figura de Yeats tanto por el inédito rigor que impuso a la poesía como por sus exploraciones en torno a la identidad Irlandesa (y, por extensión, las de sus poetas) que se nutrieron de elementos legendarios y tradicionales recuperados por el citado movimiento. Tras la firma del Tratado anglo-irlandés en 1922 se acentuó la veta política del Revival generando una amalgama, “neo–Revivalist poets were not merely influenced by official ideology; the official ideology was a form of poetry, an aestheticised mode of the anti-British struggle based on an inversion of colonial discourses” (Campbell 2003: 23). Este “revamped ruralist ideal, suitably Catholicised and purged of its English and continental taints, played a large role in the process of state formation” (ibid: 24) y en el proceso tomó distancia de Yeats ya que una sutil diferencia con el Revival original fue imponer una masculinidad gaélica a la sumisa feminidad celta. Esta tensión se advierte en la manera en que Kavanagh articuló su experiencia en contraste con la imagen idealizada del entorno rural como forma de definir una suerte de ser nacional (en la línea que va de sus primeros poemas, de fines de la década de 1920, a “The Great Hunger”, de 1942) y la abierta distancia con instancias políticas y académicas (la censura, la Universidad). Entre los primeros poemas encontramos un lirismo que se demora en la contemplación pastoral, contemplación no exenta de un cierto tono místico: “a beautiful landscape is contemplated by the poet labourer, a place of divine manifestation, where the privileged viewer may have a mystical vision” (Campbell 2003: 43). Son principalmente poemas breves, por ejemplo “Ploughman” que dio título a la primera colección publicada en 1936. El poema destaca el paisaje rural y sus habitantes, incluido el poeta que está presente en el pronombre “I” que inicia el primer verso de las dos primeras y última estrofas. No es ajeno a lo que describe, “Kavanagh’s poetry fashioned an anthropology of rural Ireland in which the poet becomes a native participant with a gift for observation” (Alderman & Blanton 2009: 97). Asimila la labor rural – arar – con la artística – pintar – y ambas, a su vez, con una tranquilidad y éxtasis místico: el trabajo es una forma de plegaria –“Like a prayer” –. La operación se pone en evidencia en la última estrofa: I find a star – lovely art In a dark sod. Joy that is timeless! O heart That knows God! Esta operación sería patrimonio, obligación, del poeta: “Sweet May is gone, and now must poets croon / The praises of a rather stupid June” (“After May”). Una parte de la contemplación de la naturaleza – y del trabajo rural – es ser consciente del cambio de estaciones, de ahí la cantidad de alusiones a los meses del año títulos y cuerpo de varios de estos primeros poemas. Otra lectura posible es pensar esta operación desde una forma de trascendencia, un intento de establecer una unión, una continuidad entre el hombre y la naturaleza en poemas como “To a Late Poplar” o “To a Blackbird”. En este último invierte el esquema de “Ploughsman”, el poeta usa la segunda persona para hacerse destinatario de las palabras del ave del título que no solo lo define como poeta sino que reconoce un vínculo mutuo – “O pagan poet, you / and I are one” – que se condensa, a partir del tercer verso, en el uso de la primera persona plural – “we are kindred” – o incluso en la aspiración mística final “Pleading for our conversion / with the Most High”. Este vínculo con la naturaleza adquiere una dimensión personal en “Poplar Memory” donde el padre muerto (“My father dreamt forests, he is dead”) prevalece, es una presencia activa (“peering”), en la contem- 37 plación que el poeta hace de los bosques, análogos a los que el padre había soñado en vida. Se diferencia así del tópico de la escritura y la lectura como una suerte de extensión de la vida más allá de la muerte, rastreable por ejemplo en la lengua inglesa hasta el Soneto XVIII de Shakespeare (“So long as men can breathe, or eyes can see / So long lives this, and this gives life to thee”). Sin embargo, no todo es una recreación idílica en un paisaje de ensueño sin angustias o conflictos. En lo que hace al paisaje o la naturaleza, en “Beech tree” describe el proceso de plantación de un árbol en febrero al que protegerá de los elementos y los animales en meses posteriores. El esfuerzo resulta fútil ya que It is August now, I have hoped, But I hope no more – My beech tree will never hide sparrows From hungry hawks. (Tal vez sea interesante notar que el “Beech tree” al que se refiere el título no es un árbol nativo de Irlanda sino una especie exótica que fue introducida y plantada extensivamente a partir el siglo XVIII y es, hasta hoy, una plaga que afecta a especies autóctonas). En el aspecto humano, un primer acercamiento se realiza en el poema “My People” donde se utiliza el diálogo para describir y diferenciar a los campesinos de la visión que un viajero citadino – “stranger” – tiene de ellos. “’Tis not so” dirá el poeta, que al igual que en “Ploughman” se incluirá entre los desdichados campesinos (“my people”, “my peasant folk”), antes de enumerar sus variadas postergaciones. Tamizadas por la religión (la mayúscula en “Pride” – pecado capital – y la referencia a la espera de un mundo donde “wrongs are righted”, por ejemplo), destaca la pobreza y la “condena” ¿divina? (“They work because Judge Want condemns the drones”) a desempeñar un trabajo manual en oposición al trabajo intelectual (“they cannot be schoolmasters”); condena frente a la que el poeta tendría una dispensa, marcada por la alternancia entre la primera y tercera personas; la diferencia entre el “They work” o “they cannot be” en lugar del esperable “we”. Una incomodidad, si se quiere, respecto a la condición de poeta entre esas gentes laboriosas con las que aparece relacionada la soledad En “Inniskeen Road: July Evening” la tranquilidad y el silencio rural devienen soledad y vacío señalados y remarcados por la ausencia de sombras y sonidos: “no shadow thrown / That might turn out a man or woman, not / A footfall tapping secrecies of stone”. Sombras y sonidos, ambos sustantivos abstractos que se oponen a las concretas bicicletas con las que otros se dirigen al baile. La imagen de la soledad del poeta se completa con la comparación con Alexander Selkirk y la idea de ser rey de “banks and stones and every blooming thing”. En forma similar, el brevísimo “Anna Quinn” presenta a modo de preguntas a Dios la idea de una soledad eterna, una vida sin amor que pasa, como la mujer del título o la luz del sol sobre el pasto (“The loveliness of life before me pass / Like Anna Quinn or sunlight on the grass?”) en contraste con la presencia post-mortem del padre en la contemplación de los bosques señalada previamente en “Poplar Memory”. El amor es ajeno al ser poeta, su destino es la soledad – “It is tragic to be a poet now / And not a lover” (“April Dusk”) –, concepto presente también en “Poet” donde se describe como un monje en una celda gris, nuevamente aislado de “The light, the laugh, the dance”. Separado de sus pares campesinos, también se distancia de otros poetas (y de la visión positiva del poeta que debe cantar de “After May”) en “The Irony of It”: I have not the fine audacity of men Who have mastered the pen Or the purse. The complexes of many slaves are in my verse. When I straighten my shoulders to look at the world boldly, I see talent coldly Damning me to stooped attrition. Mine was a beggar’s mission. To dreams of beauty I should have been born blind. I should have been content to walk behind, Watching the mirror–stones For the reflection of God’s delight: A second-hand teller of the story, A second-hand glory. It was not right That my mind should have echoed life’s overtones, That I should have seen a flower Petalled in mighty power. 38 Esta distinción entre el trabajo manual y el intelectual retoma la descripción del campesinado señalada en “My People” y concluye en “The Hired Boy”. En ese poema expresa un deseo de diferenciación, “Let me be no wiser than the dull / And leg-dragged boy” (subrayado propio), y la conciencia de que rechazar el “be wiser” implica resignarse a “To be damned and yet to live”, como el chico campesino del título. El ensueño pastoral convive entonces con esta visión negativa, de aislamiento, en la que ni la naturaleza ni la poesía llegan a conformar una opción de solaz. Las imágenes pastorales, el abundante verdor del pasto (“grass”) deviene barro (“clay”) en “The Great Hunger” pasando antes por campos oscuros (“dark fields”) y tierra rocosa gris en el poema “Stony Grey Soil”. Este último es una rememoración infantil en la que básicamente acusa a esa tierra rocosa de Monaghan de haberle arruinado la vida. En una visión mucho más oscura que la previamente desarrollada en “Monaghan Hills” (y la, en contraste, conciliatoria de “From Monaghan to the Grand Canal”, de 1959), el verde es reemplazado por el gris; los pastos por las rocas y el barro en un hábil juego de palabras que combina “clod” con “clog”. “your clod-conceived” propone, a partir de la doble significación de “clod” (“pedazo de barro” y “estúpido o “poco inteligente”), una suerte de determinismo a un destino de trabajo rural, no intelectual. De la misma manera funcionaría “You clogged the feet of my boyhood”, siendo “clod” a la vez un “zueco” (un tipo de calzado asociado a tareas rurales) de barro que se forma en los pies del niño que trabaja la tierra, y un “estorbo”, tanto en un sentido literal – la dificultad de caminar en el barro – como en un sentido abstracto – en alusión a las dificultades y carencias –. En relación con esto, creo que cabe recuperar un dato biográfico: Kavanagh dejó la escuela a los 13 años para trabajar en el campo y hacerse aprendiz en el taller de su padre, que era zapatero. Aunque otros poemas contemporáneos recuperan una infancia más positiva – “A Christmas Childhood”, “The Long Garden” o “'Primrose” –, en “Monaghan Hills” el recuerdo infantil se detiene en la forma en que las tierras de Monaghan le quitaron su juventud y alegría, dejando marcas en su frente. Contrastan los “lonely verses” del poema presente con el “first gay flight of my lyric” del pasado y los poblados que menciona al final, queridos en su juventud pero que son ahora “places of prohibition, constriction and death. The poem is a direct address to the soil, characterised not as fructive and supportive, but as manipulative and malign” (Campbell 2003: 45). El idilio tiene entonces un costado negativo, se confronta lo pastoral con lo anti–pastoral que Seamus Heaney caracterizó, respectivamente, como una representación de “idealised landscape with contented figures” en contraste con la representación de “sweat and pain and deprivation” (Citado en Campbell 2003: 42). Estos tres elementos, dispersos en los poemas “negativos” más arriba abordados, se condensan en el extenso “The Great Hunger” en la figura del campesino Patrick Maguire. El poema aparece enmarcado por los ecos bíblicos del “Clay is the word and clay is the flesh” inicial y el “apocalypse of clay” final. También como marco aparecen referencias teatrales, promediando la primera estrofa el poeta propone “We will wait and watch the tragedy to the last curtain”, esa “curtain” llegará en la sección final del poema: “Applause, applause, / The curtain falls. / Applause, applause” seguidos de “Silence, silence. The story is done”. A pesar de estas referencias al teatro, las voces no se presentan ordenadas en diálogos (como en “My People” o “The Wake of Books”) ni la expectativa que abre “wait and watch” (y que cierra “The story is done”) conlleva una progresión temporal lineal. En la primera sección se suceden desordenadamente meses, estaciones y festividades en una temporalidad abierta que contrasta con el perfectivo, a su vez enfatizado por la idea de la muerte –“Maguire was faithful to death”–. La temporalidad del poema se organiza a partir de una alternancia de aspecto – perfectivo, durativo – y saltos cronológicos; en la sección II se menciona que Maguire tenía sesenta y cinco años al morir su madre pero en la octava se recuerda “Sometime in July / When he was thirty-four or -five”. Esta construcción de una temporalidad no-cronológica se conjuga con los ciclos de la naturaleza (y, consecuentemente, del trabajo rural) para generar un matiz de ambivalencia o indeterminación – “Tomorrow is Wednesday – who cares?” –. La repetición de esos ciclos sumada a la futilidad de un trabajo rural en una 39 tierra infértil, barro (“clay”) antes que pastura (“grass”), genera una progresión en la que no hay progreso de la misma forma en que, en el aspecto familiar o social, no hay sucesión. Enfatiza así, desde lo temporal, la representación negativa de la vida de Maguire, ese encierro del que no hay escape. Esta es la clave del poema que será sintetizada en la imagen de la cabra atada en la sección XIII: el encierro en ciclos inútiles, la falta de alternativas, la desesperanza, atraviesan el poema y condicionan las relaciones de Maguire con el campo, las mujeres y, particularmente, la madre. La madre de Maguire aparece en la sección II; allí es presentada como “Wife and mother in one”, y un par de versos después el sentido de “O he loved his mother / Above all others” es neutralizado por “O he loved his ploughs”. “Above all others” destaca el lugar de la madre pero la repetición de “O he loved his” la equipara al arado. Los campesinos, a su vez, son retratados en el segundo verso de la Sección I como “mechanized scare-crows”; doblemente deshumanizados, son espantapájaros y máquinas, lo cual marca una distancia también con la idea de modernización o progreso (más adelante, en la sección IX, se afirma que “A man is what is written on the label”, el campesino queda reducido a la “etiqueta”, como un producto o una materia prima). En esa primera aparición de los campesinos se recupera también el contraste entre el trabajo rural y el intelectual, el verso “Is there some light of imagination in these wet clods?” opone la “imaginación” al “barro” lo que, a su vez, permite trazar una linea al “clod” y los “dark fields” del poema “Stony Grey Soil”. A la vez, se observa una cierta pasividad en el trabajo en tanto el componente activo no está marcado por el verbo – “move” – sino que se recupera a partir del sustantivo “machine” y por el poeta y lectores que los observan (“If we watch them”) hasta que, al final de esa primera estrofa (“Till the last soul / passively like a bag of wet clay / Rolls down the side of the hill”) nuevamente aparecen deshumanizados, como bolsas de barro húmedo que ruedan “pasivamente” montaña abajo. La misma naturaleza pierde el halo pastoral en esa estrofa introductoria – “Here crows gabble over worms and frogs / And the gulls like old newspapers are blown clear of the hedges, luckily” – lo cual no quita que, ocasionalmente en secciones posteriores, un resto místico/ pastoral echará luz sobre el lodazal: “Yet sometimes when the sun comes through a gap / These men know God the Father in a tree”. Pero solo ocasionalmente, el tono general en torno a la naturaleza es de severidad e infertilidad, incluso artística, la belleza de la naturaleza no consigue sacarle palabra a una musa muda en el final de la sección VI: He sat on the railway slope and watched the evening, Too beautifully perfect to use, And his three wishes were three stones too sharp to sit on, Too hard to carve. Three frozen idols of a speechless muse El resultado de ese silencio estaría en la sección VIII, caracterizada por la acumulación de versos repetidos – como “Sitting on a wooden gate” o “Said whatever came into his head” – que aparecen en grupos de tres. La sección está construida rítmicamente a partir de un esquema muy sencillo – que contrasta con el verso libre del resto del poema – mediante la repetición de palabras y fonemas finales: damn–sang–fat–that, head–away–spend–them etc; como el mismo poeta declara: “He didn’t care a damn / Said whatever came into his head / And inconsequently sang”. A partir de la figura de Maguire se construye un retrato brutalmente honesto de la vida campesina en el que el “sweat, pain and deprivation” que mencionaba Heaney rivaliza con los mitos pastorales del Revival. La visión del campesinado de Kavanagh está lejos de la glorificación que elaboró Yeats: Although the literal idea of a peasant is of a farm labouring person, in fact a peasant is all that mass of humanity which lives below a certain level of consciousness. They live in the dark cave of the unconscious and they scream when they see the light (citado en Kiberd 2005: 33) De hecho, señala Kiberd, se acerca al Stephen Hero de Joyce: “The glorified peasantry all seem to me as like one another as a peascod is to another peascod. They can spot a false coin but they represent no very admirable type of culture... They live a life of dull routine, the calculation of coppers, the weekly debauch and the weekly piety” (cit. en Kiberd 1996: 503). La madre – madre y esposa – 40 es retratada negativamente, es un personaje activo que manipula y subyuga a Maguire. El verso de la sección I “And he knows that his own heart is calling his mother a liar” la describe como mentirosa – sobre lo que volverá en la sección VII – a la vez que muestra la pasividad de Maguire, que lo sabe y no hace nada al respecto. La alternancia entre “he” y “his own heart” marca una escisión asimilable a la ambigüedad antes señalada. Maguire aparece dividido, la voz de la madre en la sección XI es comparada a un cuchillo que “It cut him up the middle till he became more woman than man”. Esa representación de un Maguire feminizado, castrado –idea que se extiende a otros hombres en la sección II al hablar de los campos, “Where eunuchs can be men”–, se presenta también en relación a la escritura “Nobody will ever read the wild, sprawling, scrawling mad woman’s signature, / The hysteria and the boredom of the enclosed nun of his thought” (sección IX, subrayado propio) y al envejecimiento “It was the menopause, the misery-pause” (sección XI). Hombre y eunuco, el hambre del título no alude tanto a la histórica hambruna del siglo XIX como al deseo sexual no consumado. Maguire, hijo y esposo a la vez, está casado con los campos y su relación con otras mujeres está marcada por la pasividad: en la sección VII se limita a observar a Agnes levantarse la falda y en la sección XI evita el contacto con chicas escolares por el miedo a la cárcel y la ley, miedo análogo al que produce la religión cuando, en la sección IV, “He met a girl carrying a basket … And he saw Sin”. Religión y madre formarán un tandem represor que se articula a partir del pecado: “Now go to Mass and pray and confess your sins / And you’ll have all the luck,’ his mother said”. Los únicos actos de Maguire en relación al sexo son masturbarse o darse tiempo, “he gave himself another year” (sección IX), año en el que espera una nueva vida, con matrimonio y dinero. La falta de consumación del deseo sexual excede a Maguire, se proyecta sobre los otros hombres del condado que pueden ver pero no mirar: The young women ran wild And dreamed of a child. ... But no one would take them, No man could ever see That their skirts had loosed buttons Deliberately loosed buttons. O the men were as blind as could be (sección VIII) y, a su vez, puede ser asimilada a la infertilidad del campo: In that country, in that metaphysical land, Where flesh was a thought more spiritual than music, Among the stars – out of the reach of the peasant’s hand. (sección VII) Esta infertilidad suma un aspecto a la idea de la temporalidad ambigua previamente mencionada, en lugar de progresión familiar se aprecia una especie de atemporal presente en el que el soltero(n) Maguire sigue unido “To a mother’s womb by the wind-toughened navel-cord / Like a goat tethered to the stump of a tree / He circles around and around” (Sección XIII). Estos dos versos concentran la idiosincrasia de Maguire – y, por extensión, al campesinado –. En principio, la dependencia de la madre en la imagen del cordón umbilical que, además, junto con “womb” remite a un estado embrionario de desarrollo donde la dependencia es mucho más estrecha, una simbiosis. El verso siguiente permite reponer una relación similar con la granja y el trabajo rural. Aparece atado a un “tocón” (“stump”), un resto de árbol; esta representación difiere de los árboles en poemas previos, asociados con la vida y una visión positiva de la naturaleza y la relación que el hombre tiene con ella. Al mismo tiempo, “Stump” puede pensarse como “muñón” y, en ese sentido, también podría relacionarse con la vida truncada de Maguire o, directamente, a su castración. En el mismo verso Maguire aparece animalizado a partir de la comparación con la cabra atada. Los círculos que traza se asocian a los ciclos de la naturaleza que, a su vez, determinan los del trabajo rural conformando una temporalidad donde el progreso no necesariamente implica progresión: se avanza pero no se llega a ninguna parte, no hay salida de ese círculo, lo que llevará a Maguire a clamar al Cielo en el último verso de la Sección XII: “O Christ! I am locked in a stable with pigs and cows for ever”. El poema presenta una doble negatividad en el retrato de Maguire, no solo la vida es gris y sin sentido (“It means nothing / Not a damn thing”, sección V) sino que, en esa escisión 41 señalada antes, es consciente de la situación pero no hace nada, porque no puede hacer nada: “Is there nothing he can do? / Is there no escape? / No escape, no escape” (sección XIII). Tampoco encuentra solaz en elementos naturales o místicos. La naturaleza es sinónimo de trabajo, son escasas las apariciones del verde, la luz solar o el agua. Dios aparece como fundamento, causa (o excusa) de la situación: “the grief and defeat of men like these peasants / Is God’s way – maybe –” (sección XI). Esta determinación religiosa aparece en la sección VIII en la idea de la vida como “purgatorio”, como un “mal necesario”: And Patrick Maguire From his purgatory fire Called the gods of the Christian to prove That this twisted skein Was the necessary pain And not the rope that was strangling true love Al igual que Maguire, Dios aparece escindido “God is not all / In one place, complete” (sección VI) y no responde. En un poema previo, “Why Sorrow?” el padre Mat dice: Love, happiness, riches, Christ cannot give. He is the bitter-tasted, wrong-turned. You will get From Christ if you pray for love a laugh too late, And riches Christly-come will be desire Without escape for it En esa línea, el consuelo que Dios ofrece(ría) es un consuelo final: “Christ will meet them at the end of the world, the slow and speedier. / But the fields say: only Time can bless” (sección IV), similar al “await the world where wrongs are righted” de “My People”. El paradójico “slow and speedier” nuevamente instala esa temporalidad ambigua, circular, en la cual “only Time can bless” parece irónico. Las mayúsculas permiten asociar “Time” con “Christ” en una relación mediada por los “fields” pero el adversativo “but” marca una distancia, como si los campos – a pesar de la minúscula – fueran los que tienen la última palabra. Naturaleza e Iglesia aparecen asociadas en la presión negativa que ejercen sobre los campesinos: Life dried in the veins of these women and men: The grey and grief and unlove, The bones in the backs of their hands, And the chapel pressing its low ceiling over them. (sección IX) Habría en los poemas de Kavanagh una diferencia entre Dios o Cristo y la Iglesia o, mejor, una consciencia de la forma en que la Iglesia en tanto institución utiliza a Dios para ejercer un control represivo (la “chapel pressing its low ceiling” de los versos previos). Maguire depositaría una cierta confianza en Dios a pesar de reconocerlo como instaurador del orden que debe padecer, mientras que con la Iglesia la relación está teñida de sospecha. El padre Mat del poema “Why Sorrow?” antes mencionado vive con miedo de que alguien le escriba al Obispo contándole “escándalos” o la forma en que “worshipped more the flowers in the garden / Than God”, verso en el que queda expresada la distancia entre el sacerdote y Dios. En “The Great Hunger”, la Iglesia se asocia al control represivo de la madre y la naturaleza, en la sección VII la madre ordena a Maguire ir a misa y confesar sus pecados y mientras ella “was setting up the lie / She trusted in Nature that never deceives … Religion’s walls expand to the push of nature”. Este contexto condiciona el desarrollo personal y comunal. Maguire, que no puede hablar con mujeres por miedo (a la ley, a la religión, a la madre), habla solo o con los caballos: “One talks to the horse as to a brother” (sección XIV) lo que remarca la soledad, la animalización y la descomposición de la familia como unidad (lo cual ya estaba instalado a partir de la relación con la madre). Las charlas de los campesinos son completamente banales, así como las lecturas (cfr. secciones IX y X), la comunidad rural también aparece descompuesta, marcada por los celos y la rivalidad – “From every second hill a neighbour watches / With all the sharpened interest of rivalry” (sección III) –. Un ambiente completamente distinto al que el Revival y la propaganda política de la nueva república intentaron construir, la “Irlanda ideal” que Éamon de Valera describió en su famoso discurso del 17 de marzo de 1943, “The Ireland that we dreamed of”: 42 The ideal Ireland that we would have, the Ireland that we dreamed of, would be the home of a people who valued material wealth only as a basis for right living, of a people who, satisfied with frugal comfort, devoted their leisure to the things of the spirit – a land whose countryside would be bright with cosy homesteads, whose fields and villages would be joyous with the sounds of industry, with the romping of sturdy children, the contest of athletic youths and the laughter of happy maidens, whose firesides would be forums for the wisdom of serene old age. The home, in short, of a people living the life that God desires that men should live (De Valera & Moynihan 1980: 466). La distancia de Kavanagh a estos supuestos llega a su punto más alto (y corrosivo) en la sección XIII de “The Great Hunger”. Enmarcada por la “mirada” del “mundo” y los “viajeros”, comienza con una exposición que resume los supuestos sobre el campesino, “fuente” de la cultura y la religión, de poetas y músicos: The peasant has no worries; In his little lyrical fields He ploughs and sows; He eats fresh food, He loves fresh women, He is his own master; As it was in the Beginning, The simpleness of peasant life. The birds that sing for him are eternal choirs, Everywhere he walks there are flowers. His heart is pure, His mind is clear, He can talk to God as Moses and Isaiah talked – The peasant who is only one remove from the beasts he drives. para luego destruir dichos supuestos en un crescendo que culmina en la imagen de la cabra atada antes mencionada, que es “the weak, washy way of true tragedy”. La clave de esta operación está dada por los versos que enmarcan esa primera exposición: el mundo y los viajeros que “miran” al campesino, retomando el esquema de “My People” donde el que inquiere al poeta sobre los campesinos es un “Stranger”, “extranjero”, “extraño”. Kavanagh opera un cambio de paradigma, cambia el mirar “desde fuera” por un mirar “desde adentro”: Kavanagh’s tirades against the poetry and ideology of the Revival were spoken from the viewpoint of a farmer–poet dismayed by writers of national pastoral who, in many cases, had little genuine experience of rural, let alone agricultural life (Campbell 2003: 43). Parte de ese cambio de paradigma es anteponer al provincialismo el parroquialismo. Pensar unidades pequeñas y cohesivas de comunidad – las parroquias – a las que considera “base” de todas las “grandes civilizaciones”, el “Parochialism is universal: it deals with the fundamentals” (cfr. O’Grady 1996), por lo que no es extraño que en “Epic” asimile una disputa territorial local al tratado de 1938 sobre Checoslovaquia. Homero, ante la pregunta de qué es más importante, responde que construyó la Iliada a partir de “A local row”. En los poemas que recorrimos, “The Great Hunger” pero también otros como “My People”, “The Hired Boy” o “The Irony of It”, se aprecia cómo la voz de Kavanagh parece construirse a partir del contraste entre dos paradigmas: la ciudad y el campo, la educación y la experiencia, los poetas y los campesinos. Siguiendo a Seamus Heaney, el aporte de Kavanagh a la poesía Irlandesa es unir esos paradigmas: Kavanagh’s genius had achieved singlehanded what I and my grammar-schooled, arts-degreed generation were badly in need of – a poetry which linked the small farm life which produced us with the slim-volum world we were now supposed to be fit for (1988: 9). No es, sin embargo, una intervención conciliatoria que se limita a unir. La inconformista voz lírica de Kavanagh deconstruye los paradigmas del Revival y los nacionalistas, revelando un cultura provincial construida e impuesta desde la comodidad de la ciudad. La crudeza con que ese inconformismo se manifiesta en los poemas está lejos de la pose o la afectación: We might say that lyric celebration was to Kavanagh what witty expression was to Oscar Wilde – in the beginning, a matter of temperament, a habit of style, a disposition of the artist’s fundamental nature, but, in the end, a matter of redemptive force, a resource that maintained the artist’s inner freedom in the face of worldly disappointments, an infrangible dignity (Heaney 1988: 12). Esa libertad y dignidad inquebrantable dan sustento a la voz de Kavanagh tanto en la pintura 43 áspera de la vida rural de “The Great Hunger” como en el contraste satírico con el Revival de “Yeats” y están en la base de su poética inconformista: No System, no Plan, Yeatsian invention, No all-over Organizational prover. Let words laugh And people be stimulated by our stuff (“Mermaid Tavern”) Bibliografía Alderman, Nigel & Blanton, C.D. 2009. A Concise Companion to Postwar British and Irish Poetry. Malden, MA: Wiley-Blackwell. Campbell, Matthew. 2003. The Cambridge Companion to Contemporary Irish Poetry. Cambridge: Cambridge University Press. Crotty, Patrick. 1995. Modern Irish Poetry. Belfast: Blackstaff Press. De Valera, Éamon & Moynihan, Maurice (ed.). 1980. “The Ireland that we dreamed of” en Speeches and Statements by Eamon De Valera: 1917-73. Dublin: Gill & Macmillan. 466-469. Rafroidi, Patrick. 1987. “The sense of place in Seamus Heaney’s poetry”, en Genet, J. 1987. Studies on Seamus Heaney. Caen: Centre de publications de l’Université de Caen. 79–88. (disponible en: http://books.openedition.org/puc/201 [consultado: 20/10/2014] Gillis, Alan. 2005. Irish Poetry of the 1930s. Oxford: Oxford University Press. Heaney, Seamus. 1988. “The Placeless Heaven: Another Look at Kavanagh” en The Government of the Tongue: The 1986 T.S. Eliot Memorial Lectures, and Other Critical Writings, London: Faber, 3-14. Reeditado en Finders Keepers. New York: Farrar, Straus and Giroux, 2002: 146-157. Kavanagh, Patrick. 1959. “From Monaghan to the Grand Canal”. Studies: An Irish Quarterly Review Vol. 48 No. 189, 29–36. http://www.jstor.org/stable/30103526 [consultado: 20/10/2014] _______. 2003. A Poet’s Country. Ed Antoinette Quinn. Dublin: Lilliput Press. _______. 2005. Collected Poems. Ed. Antoinette Quinn. Londres: Penguin UK. Kiberd, Declan. 1996. Inventing Ireland. Londres: Vintage. _______. 2005. The Irish Writer and the World. Cambridge: Cambridge University Press. O’Grady, Thomas. 1996. “The Parish and the Universe: A Comparative Study of Patrick Kavanagh and William Carleton” en Studies: An Irish Quarterly Review, Vol. 85 No. 337, 17–26. http://www.jstor.org/stable/30092026 [consultado: 20/10/2014] Received 20th November 2014 Last version 21st January 2015 Maximiliano Brina es Licenciado en Letras por la Universidad de Buenos Aires donde también cursó la Maestría en Estudios Literarios. Su tema de tesis versa sobre la producción poética española contemporánea.	609
https://openalex.org/W2044263391	OpenAlex	Open Science	CC-By	2,015	The putative Na+/Cl−-dependent neurotransmitter/osmolyte transporter inebriated in the Drosophila hindgut is essential for the maintenance of systemic water homeostasis	Lang Zhuo	English	Spoken	8,070	13,564	SUBJECT AREAS: HOMEOSTASIS BEHAVIOURAL GENETICS Received 7 October 2014 Accepted 29 December 2014 Published 23 January 2015 Received 7 October 2014 Accepted 29 December 2014 Published 23 January 2015 Zhuo Luan, Caitlin Quigley & Hong-Sheng Li Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA 01605, USA. Most organisms are able to maintain systemic water homeostasis over a wide range of external or dietary osmolarities. The excretory system, composed of the kidneys in mammals and the Malpighian tubules and hindgut in insects, can increase water conservation and absorption to maintain systemic water homeostasis, which enables organisms to tolerate external hypertonicity or desiccation. However, the mechanisms underlying the maintenance of systemic water homeostasis by the excretory system have not been fully characterized. In the present study, we found that the putative Na1/Cl2-dependent neurotransmitter/ osmolyte transporter inebriated (ine) is expressed in the basolateral membrane of anterior hindgut epithelial cells. This was confirmed by comparison with a known basolateral localized protein, the a subunit of Na1-K1 ATPase (ATPa). Under external hypertonicity, loss of ine in the hindgut epithelium results in severe dehydration without damage to the hindgut epithelial cells, implicating a physiological failure of water conservation/absorption. We also found that hindgut expression of ine is required for water conservation under desiccating conditions. Importantly, specific expression of ine in the hindgut epithelium can completely restore disrupted systemic water homeostasis in ine mutants under both conditions. Therefore, ine in the Drosophila hindgut is essential for the maintenance of systemic water homeostasis. Correspondence and requests for materials should be addressed to Z.L. (Zhuo.Luan@ umassmed.edu) or H.-S.L. (Hong-Sheng. Li@umassmed.edu) W b W ater homeostasis is essential for the survival of all organisms. The mammalian kidney and the Malpighian tubule and hindgut of insects play indispensable roles in maintaining water homeostasis over a wide range of external or dietary osmolarities. These organs can increase water conservation and absorption to maintain systemic water homeostasis, which enables organisms to tolerate external hypertonicity or desiccation1. The mammalian kidney regulates water balance mainly through the antidiuretic hormone (ADH)2–5, which enhances water absorption. Failure of antidiuretic mechanisms can result in disrupted systemic water homeostasis, causing pathological conditions like Diabetes Insipidus6. Although antidiuretic factors for the enhancement of water absorption, such as Schgr-ITP and CAPA-related peptides, are also present in insects7–17, the mechanisms of water conservation and absorption in the excretory system are not fully characterized, especially in Drosophila. Correspondence and requests for materials should be addressed to Z.L. (Zhuo.Luan@ umassmed.edu) or H.-S.L. (Hong-Sheng. Li@umassmed.edu) SUBJECT AREAS: HOMEOSTASIS BEHAVIOURAL GENETICS Previous studies have shown that loss of the putative Na1/Cl2-dependent neurotransmitter/osmolyte trans- porter inebriated (ine) causes hypersensitivity to dietary hypertonicity in Drosophila; however, the mechanism underlying this effect remains unknown18. Ine is a member of the Na1/Cl2-dependent neurotransmitter/osmo- lyte transporter family, which is conserved across invertebrates and vertebrates18,19. Members of this family share several common structural features, including 12 transmembrane domains flanked by intracellular N and C termini, and an extracellular loop between the third and fourth transmembrane domains20. These proteins play critical roles in neurotransmission, as well as cellular and systemic homeostasis, by transporting neurotransmit- ters, osmolytes, and energy metabolites across the plasma membrane. There is sequence similarity between ine and the betaine/GABA transporter (BGT1), a mammalian member of the Na1/Cl2-dependent neurotransmitter/ osmolyte transporter family. Both BGT1 and ine are expressed in the central nervous system (CNS), as well as organs that perform water absorption, and both are involved in the control of neuronal excitability and tolerance SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 1 www.nature.com/scientificreports (Fig. 2G). We labeled all membranes of hindgut epithelial cells by driving membrane-bound GFP with hindgut-Gal4, and the basolat- eral membrane with anti-ATPa antibody. Upon co-staining with anti-ine antibody, we found that ine completely co-localized with ATPa in the basolateral membrane of the hindgut epithelium (Fig. 2C and D). BGT1 also localizes to the basolateral membrane of renal medullary cells, which allows the cells take up betaine from circulation rather than the medullary lumen26. Similarly, ine might transport an as yet unknown osmolyte into hindgut epithelial cells from the hemolymph, rather than the hindgut lumen. to hypertonicity18,19,21–23. This suggests that these two proteins may function through a similar mechanism. Betaine, an active organic compound, is the substrate of BGT1 in renal medullary cells; how- ever, the substrate of ine has yet to be identified. Betaine, like other intracellular organic osmolytes, can protect cells from external hypertonicity by balancing high extracellular osmolarity and pre- serving cell volume without interfering with cell function24,25. However, no direct genetic evidence supports the osmoprotective function of the BGT1-mediated accumulation of betaine in renal medullary cells26,27. Specifically, BGT1 knockout mice are healthy, and renal medullary cells appear to be normal in the hypertonic environment of the renal medulla26. Therefore, the physiological function of the Na1/Cl2-dependent neurotransmitter/osmolyte transporter in the excretory system remains to be elucidated. Results Ine is expressed in the basolateral membrane of adult hindgut epithelial cells and co-localizes with Na1-K1 ATPase. Although ine mRNA is observed in the hindgut and Malpighian tubules of Drosophila embryos via whole-mount in situ hybridization18,19, the expression pattern of ine protein in the adult fly is still uncharacterized. To answer this question, we generated an anti-ine antibody to observe the subcellular localization of ine, and hindgut- Gal4 to label hindgut epithelial cells (Fig. 1). The hindgut is divided into two sections: anterior (the ileum) and posterior (the rectum). We performed double-immunofluorescent staining on the gut and Malpighian tubules using antibodies against b-alanine, which generally labels the structure of the gut, and ine. We found that ine is specifically expressed in the basolateral membrane of the anterior hindgut epithelium, but not in other parts of the hindgut or in the Malpighian tubules (Fig. 2A, B and E)28. This expression pattern conflicts with previous reports of ine mRNA distribution29; how- ever, the discrepancies may be due to various biological factors such as complex gene regulatory mechanisms30. Ine is not involved in the osmoprotective response to external hypertonicity in anterior hindgut epithelial cells. Huang et al. postulated that elevated intracellular levels of Na1 and K1 in hindgut epithelial cells in response to external hypertonicity would be lethal, either through a necrotic or apoptotic mechanism, unless normal intracellular Na1 and K1 levels could be restored. They proposed an osmoprotective role for ine, in which an osmolyte transported by ine increased intracellular molality thus allowing The subcellular localization of ine was further confirmed by com- parison with the a subunit of Na1-K1 ATPase (ATPa), which is known to localize to the basolateral membrane in Malpighian tubules31. We observed that ATPa is also localized to the basolateral membrane of the hindgut epithelium using an anti-ATPa antibody Figure 1 \| The hindgut-specific expression pattern of the hindgut-Gal4 line as visualized using UAS-GFP. In pupae, GFP signal is detected in the posterior part (B) but not in the anterior part (A). The GFP signal is detected in the abdomen of the adult fly (C). The hindgut-Gal4 is specifically expressed in the hindgut but not in the midgut and Malpighian tubules (D and E). Scale bars: 100 mm. Figure 1 \| The hindgut-specific expression pattern of the hindgut-Gal4 line as visualized using UAS-GFP. SUBJECT AREAS: HOMEOSTASIS BEHAVIOURAL GENETICS Ine in the hindgut epithelium is essential for tolerance of dietary hypertonicity in Drosophila. Previous studies have shown that loss of ine causes hypersensitivity to dietary hypertonicity in Drosophila. We sought to repeat these findings. To characterize the differential tolerance of dietary hypertonicity between WT flies and ine mutants, we prepared fly food media with a 0.2 M salt solution in place of water. Consistent with previous findings18, we observed a sensitivity to dietary hypertonicity in ine mutants. We studied flies bearing two different mutations in the ine gene, ine2 and ine3, and found in both cases that flies maintained on normal medium exhibited no lethality, whereas those maintained on hypertonic media died within 10 days. In contrast, dietary hypertonicity had no effect on the viability of WT flies (Fig. 3C). Because ine is expressed in the CNS as well as the hindgut, we tested whether the intolerance to dietary hypertonicity was due to the loss of ine specifically in the CNS or the hindgut tissue. Ine has 2 isoforms, RA and RB, which may have different functions. We rescued the ine2 and ine3 mutant phenotypes by overexpressing either the RA or RB isoform using hindgut-Gal4. Overexpression of either isoform resulted in localization of the protein to the basolateral membrane (Fig. 4A), similar to the endogenous distribution pattern (Fig. 2). This result suggests that the overexpressed protein functions normally. Both the RA and RB isoform were sufficient to rescue lethality in ine2 and ine3 flies maintained on hypertonic media. However, expression of either the RA or RB isoform in neurons or glia using elav- and repo-Gal4, respectively, did not rescue lethality in mutants fed on hypertonic media (Fig. 4C). These results indicate that ine is required in the hindgut epithelium, but not the CNS, for tolerance to dietary hypertonicity. By investigating the function of ine in Drosophila, an excellent genetic model in which gene expression can be evaluated and manipulated in vivo, we may begin to understand the physiological function of Na1/Cl2-dependent neurotransmitter/osmolyte trans- porters, including BGT1, in the excretory system. In this study, we elucidate the role of ine in the Drosophila hindgut, and reveal a novel mechanism mediated by ine for the maintenance of systemic water homeostasis. Results In pupae, GFP signal is detected in the posterior part (B) but not in the anterior part (A). The GFP signal is detected in the abdomen of the adult fly (C). The hindgut-Gal4 is specifically expressed in the hindgut but not in the midgut and Malpighian tubules (D and E). Scale bars: 100 mm. SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 2 www.nature.com/scientificreports Figure 2 \| Ine is localized at the basolateral membrane of the hindgut epithelial cells. The hindguts were stained with an anti-ine antibody (red). (A) and (B), both the hindgut epithelium (hg) and visceral muscle layer (vm) were labeled with a b-alanine antibody (green). Ine localizes to the basolateral membrane, but not the apical membrane, of the anterior hindgut epithelial cells. (C) and (D), the hindguts of hindgut-Gal4 . UAS-GFP flies were stained with an anti-ine antibody (red) and an antibody against the a subunit of Na1-K1 ATPase (ATPa, blue). ATPa signal co-localizes with ine in the hindgut epithelial cells. (E), Ine is not expressed in the Malpighian tubules. (F) and (G), ATPa is localized to the basolateral membrane of the hindgut and Malpighian tubules. Scale bars: a, c, e and f, 100 mm; b, d and g, 50 mm. Fi 2 \| I i l li d t th b l t l b f th hi d t ith li l ll Th hi d t t i d ith ti i tib d ( d) (A) d Figure 2 \| Ine is localized at the basolateral membrane of the hindgut epithelial cells. The hindguts were stained with an anti-ine antibody (red). (A) and (B), both the hindgut epithelium (hg) and visceral muscle layer (vm) were labeled with a b-alanine antibody (green). Ine localizes to the basolateral membrane, but not the apical membrane, of the anterior hindgut epithelial cells. (C) and (D), the hindguts of hindgut-Gal4 . UAS-GFP flies were stained with an anti-ine antibody (red) and an antibody against the a subunit of Na1-K1 ATPase (ATPa, blue). ATPa signal co-localizes with ine in the hindgut epithelial cells. (E), Ine is not expressed in the Malpighian tubules. (F) and (G), ATPa is localized to the basolateral membrane of the hindgut and Malpighian tubules. Scale bars: a, c, e and f, 100 mm; b, d and g, 50 mm. media for 4 days, after which hindguts were dissected out and stained with Trypan Blue. Results When maintained on normal medium, ine3 flies had a similar hemolymph volume and total body water content to WT flies (Fig. 3B). Under external hypertonicity, the hemolymph volume and total body water content of the ine3 flies declined dramatically while those of WT flies were not affected. Overexpression of the RA or RB isoform in the hindgut epithelium by hindgut-Gal4 completely and independently rescued the severe loss of body water in mutants (Fig. 4B), indicating that the two isoforms have similar functions. These results suggest that ine in the hindgut epithelium may mediate water conservation/ absorption, which is essential for the maintenance of systemic water homeostasis under external hypertonicity. In humans, losing approximately 20% of the body’s water content is known to cause delirium, coma and death34,35. Therefore, the severe dehydration caused by the failure of water conservation/absorption may be the primary reason for the death of ine mutants under external hypertonicity. water content in WT, ine3, and mutant flies rescued with either the RA or RB isoform of ine. Adult flies were maintained on normal or hypertonic media for 4 days, after which hemolymph volume and total body water content of individual flies were quantified. When maintained on normal medium, ine3 flies had a similar hemolymph volume and total body water content to WT flies (Fig. 3B). Under external hypertonicity, the hemolymph volume and total body water content of the ine3 flies declined dramatically while those of WT flies were not affected. Overexpression of the RA or RB isoform in the hindgut epithelium by hindgut-Gal4 completely and independently rescued the severe loss of body water in mutants (Fig. 4B), indicating that the two isoforms have similar functions. These results suggest that ine in the hindgut epithelium may mediate water conservation/ absorption, which is essential for the maintenance of systemic water homeostasis under external hypertonicity. In humans, losing approximately 20% of the body’s water content is known to cause delirium, coma and death34,35. Therefore, the severe dehydration caused by the failure of water conservation/absorption may be the primary reason for the death of ine mutants under external hypertonicity. that the ine mutants were more sensitive than WT flies to desiccation, indicating that the rate of water loss was higher in ine mutants than in WT flies. Drosophila lose water through three mechanisms: excretion from the mouthparts and anus, cuticular transpiration, and respir- atory loss through the spiracles36. Results Both WT and ine3 flies exhibited little or no Trypan Blue staining in the anterior hindgut (Fig. 5B and C), indicating that without ine, anterior hindgut epithelial cells are not damaged by external hypertonicity, and that the osmoprotective response of the epithelial cells against external hypertonicity is normal. Therefore, ine does not function as an osmoprotector in anterior hindgut epithelial cells. We propose that ine has a direct, physiological role in water conservation/absorption that is not secondary to protection of the hindgut epithelium from damage. Na1 and K1 to move out of the cell, and returning cell volume and ion concentration to normal physiological levels18. If this osmoprotective theory is correct, hindgut epithelial cells without ine would undergo necrotic or apoptotic cell death under conditions of external hyper- tonicity. Therefore, we examined whether anterior hindgut epithelial cells were damaged by external hypertonicity in the absence of ine. We labeled hindgut epithelial cells with GFP using hindgut-Gal4 in a WT or ine3 background and maintained the flies on normal or hypertonic media. After 4 days, we dissected out the hindgut and were able to detect GFP signal in the hindgut epithelium. This result demonstrates that external hypertonicity does not affect GFP expression in the hindgut epithelial cells of WT or mutant flies (Fig. 5A), and indicates that epithelial cells in those flies were healthy. To further examine tissue damage, including necrotic and apoptotic cell death, we stained the hindgut with Trypan Blue, a dye that is excluded from intact cells but is rapidly absorbed by dead or dying cells32. In normal conditions, WT gut showed minimal staining in the ileum and moderate staining in the rectum, which may be due to desiccation damage of remnants of muscle, fat, and connective tissue surrounding it. WT and ine3 adult flies were maintained on normal or hypertonic The expression of ine in hindgut epithelium is indispensable for the maintenance of systemic water homeostasis. The hindgut is important for fluid absorption in many insects33; however, this function has never been demonstrated in the hindgut of adult Drosophila. Considering its specific expression in the hindgut and the hypersensitivity of mutants to dietary hypertonicity, we hypothesized that ine in the hindgut epithelium is essential for water conservation/absorption in response to external hypertonicity. Results Therefore, we examined the volume of hemolymph and the total body SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 3 Figure 3 \| The loss of ine in the hindgut epithelium disrupts systemic water homeostasis, which leads to death under conditions of external hypertonicity. (A), the ine signal is completely abolished in the hindgut epithelium of the null mutant ine3. Scale bars: 100 mm. (B), when maintained on normal medium, ine3 mutants have a similar hemolymph volume and total body water content to the WT flies. However, the hemolymph volume and total body water content of ine3 mutants declined dramatically, while those of the WT flies were not affected, when the flies were maintained on hypertonic media (n 5 5, t-tests, two tails). Each bar represents the mean 6 S.E.M. Asterisk (): p , 0.01. (C), ine mutant flies exhibited no lethality when maintained for up to 10 days on normal medium, while those maintained on hypertonic media died within 10 days. Dietary hypertonicity does not affect WT flies. Each point represents the mean 6 S.E.M. The data are representative of three independent experiments. Figure 3 \| The loss of ine in the hindgut epithelium disrupts systemic water homeostasis, which leads to death under conditions of external hypertonicity. (A), the ine signal is completely abolished in the hindgut epithelium of the null mutant ine3. Scale bars: 100 mm. (B), when maintained on normal medium, ine3 mutants have a similar hemolymph volume and total body water content to the WT flies. However, the hemolymph volume and total body water content of ine3 mutants declined dramatically, while those of the WT flies were not affected, when the flies were maintained on hypertonic media (n 5 5, t-tests, two tails). Each bar represents the mean 6 S.E.M. Asterisk (): p , 0.01. (C), ine mutant flies exhibited no lethality when maintained for up to 10 days on normal medium, while those maintained on hypertonic media died within 10 days. Dietary hypertonicity does not affect WT flies. Each point represents the mean 6 S.E.M. The data are representative of three independent experiments. water content in WT, ine3, and mutant flies rescued with either the RA or RB isoform of ine. Adult flies were maintained on normal or hypertonic media for 4 days, after which hemolymph volume and total body water content of individual flies were quantified. Results Water conservation/absorption mediated by ine may reduce the rate of water loss through excretion to combat dehydration. The expression of either the RA or RB iso- form of ine in the hindgut epithelium by hindgut-Gal4, but not by repo- or elav-Gal4, completely rescued the sensitivity of mutants to desiccation. In contrast, under hypotonic conditions with only a water supply, mutants and WT flies exhibited a similar resistance to starvation. These results indicate that ine is indispensable for water conservation/absorption under conditions of desiccation but not starvation, and is essential for the maintenance of systemic water homeostasis. Discussion h d Each bar represents the mean 6 S.E.M. (C), overexpression of the RA or RB subunit of ine exclusively in hindgut epithelial cells completely rescued the lethality of ine2 or ine3 mutants maintained on hypertonic media. Each point represents the mean 6 S.E.M. The data are representative of three independent experiments. igure 4 \| Specific overexpression of ine in the hindgut epithelium can restore disrupted systemic water homeostasis and rescue lethality under d ypertonicity in ine mutants. (A), overexpression of the RA or RB isoforms of ine using hindgut-Gal4 results in targeting to the basolateral mem f the hindgut epithelial cells in the ine3 mutant. Scale bars: a and b, 200 mm; a9 and b9, 100 mm. (B), overexpression of the RA or RB isoforms of ine i d i h li l l h h l h l d l b d f i 3 fli l l l ( Figure 4 \| Specific overexpression of ine in the hindgut epithelium can restore disrupted systemic water homeostasis and rescue lethality under dietary hypertonicity in ine mutants. (A), overexpression of the RA or RB isoforms of ine using hindgut-Gal4 results in targeting to the basolateral membrane of the hindgut epithelial cells in the ine3 mutant. Scale bars: a and b, 200 mm; a9 and b9, 100 mm. (B), overexpression of the RA or RB isoforms of ine in the hindgut epithelium can completely restore the hemolymph volume and total body water content of ine3 flies to normal levels (n 5 5, t-tests, two tails). Each bar represents the mean 6 S.E.M. (C), overexpression of the RA or RB subunit of ine exclusively in hindgut epithelial cells completely rescued the lethality of ine2 or ine3 mutants maintained on hypertonic media. Each point represents the mean 6 S.E.M. The data are representative of three independent experiments. homeostasis in response to fluctuations in external osmotic condi- tions7–10. However, in adult Drosophila, the water conservation/ absorption mechanisms of the hindgut have not been elucidated. Our results demonstrate that ine is expressed in the basolateral mem- brane of the hindgut epithelium, suggesting that ine transports sub- strate from the hemolymph into hindgut epithelial cells. Surprisingly, under conditions of external hypertonicity, the systemic water home- ostasis of ine mutant flies is disrupted, whereas that of WT flies is not disturbed. Discussion h d We have demonstrated that the mediation of water conservation/ absorption by ine in the hindgut is essential for the maintenance of systemic water homeostasis in Drosophila. In insects, systemic water homeostasis is tightly regulated by the excretory system, including the Malpighian tubules and the hindgut, to ensure a constant internal environment37. The dynamic balance between Malpighian tubule secretion and hindgut reabsorption, both of which are controlled by diuretic and antidiuretic hormones or factors, maintains water We questioned whether the mechanism mediated by ine functions under conditions of other than hypertonicity, such as desiccation, in which water is withheld, and starvation, in which flies are only given a water supply. To address this question, we measured the resistance of ine mutant flies to desiccation and starvation (Fig. 6). We found SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 4 Figure 4 \| Specific overexpression of ine in the hindgut epithelium can restore disrupted systemic water homeostasis and rescue lethality under dietary hypertonicity in ine mutants. (A), overexpression of the RA or RB isoforms of ine using hindgut-Gal4 results in targeting to the basolateral membrane of the hindgut epithelial cells in the ine3 mutant. Scale bars: a and b, 200 mm; a9 and b9, 100 mm. (B), overexpression of the RA or RB isoforms of ine in the hindgut epithelium can completely restore the hemolymph volume and total body water content of ine3 flies to normal levels (n 5 5, t-tests, two tails). Each bar represents the mean 6 S.E.M. (C), overexpression of the RA or RB subunit of ine exclusively in hindgut epithelial cells completely rescued the lethality of ine2 or ine3 mutants maintained on hypertonic media. Each point represents the mean 6 S.E.M. The data are representative of three independent experiments. www.nature.com/scientificreports Figure 4 \| Specific overexpression of ine in the hindgut epithelium can restore disrupted systemic water homeostasis and rescue lethality under dietary hypertonicity in ine mutants. (A), overexpression of the RA or RB isoforms of ine using hindgut-Gal4 results in targeting to the basolateral membrane of the hindgut epithelial cells in the ine3 mutant. Scale bars: a and b, 200 mm; a9 and b9, 100 mm. (B), overexpression of the RA or RB isoforms of ine in the hindgut epithelium can completely restore the hemolymph volume and total body water content of ine3 flies to normal levels (n 5 5, t-tests, two tails). Discussion h d These results demonstrate that hindgut expression of ine mediates water conservation/absorption under external hypertoni- city and maintains systemic water homeostasis. These results also suggest possible mechanism for ine function: transport of an osmo- lyte by ine into the hindgut epithelium increases intracellular mol- arity, which enhances water conservation/absorption from the hindgut lumen. Such a function would be particularly important in the condition of external hypertonicity, when increased molality in the hindgut lumen prevents osmotic flow of water into hindgut epithelium. It could be argued that ine functions through an osmoprotective mechanism, in which increased intracellular accumulation of osmo- SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 5 www.nature.com/scientificreports gure 5 \| External hypertonicity does not damage anterior hindgut epithelial cells in the absence of ine. (A), hindgut epithelial cells were labele th GFP in WT or ine3 backgrounds using hindgut-Gal4. Maintenance on hypertonic media for 4 days did not affect the expression of GFP in th ndgut epithelial cells of WT or ine3 flies, which indicates that the cells are intact. (B), hindgut tissue of WT and ine3 flies maintained on normal pertonic media for 4 days presented minimal Trypan blue staining (C) quantification of Trypan blue staining in the anterior hindgut No signifi gure 5 \| External hypertonicity does not damage anterior hindgut epithelial cells in the absence of ine. (A), hindgut epithelial cells were label th GFP in WT or ine3 backgrounds using hindgut-Gal4. Maintenance on hypertonic media for 4 days did not affect the expression of GFP in t Figure 5 \| External hypertonicity does not damage anterior hindgut epithelial cells in the absence of ine. (A), hindgut epithelial cells were labeled with GFP in WT or ine3 backgrounds using hindgut-Gal4. Maintenance on hypertonic media for 4 days did not affect the expression of GFP in the hindgut epithelial cells of WT or ine3 flies, which indicates that the cells are intact. (B), hindgut tissue of WT and ine3 flies maintained on normal or hypertonic media for 4 days presented minimal Trypan blue staining. (C), quantification of Trypan blue staining in the anterior hindgut. No significant differences were observed in the viability of the anterior hindgut between WT and ine3 flies maintained on normal or hypertonic media (n 5 5, t-tests, two tails). Each bar represents the mean 6 S.E.M. Scale bars: 100 mm. epithelial cells against lethality under external hypertonicity26. Discussion h d The expression of several genes, including some organic transporters, is up-regulated in the hindgut in response to external hypertonicity38, supporting this possibility. lytes mediated by ine protects the hindgut epithelium from cellular death due to extracellular hypertonicity. However, we demonstrate that anterior hindgut epithelial cells are not damaged by external hypertonicity in the absence of ine, suggesting that ine function in water conservation/absorption is not secondary to an osmoprotec- tive effect. We propose the existence of other osmolytes or transpor- ters that function as osmoprotectors, and protect anterior hindgut lytes mediated by ine protects the hindgut epithelium from cellular death due to extracellular hypertonicity. However, we demonstrate that anterior hindgut epithelial cells are not damaged by external hypertonicity in the absence of ine, suggesting that ine function in water conservation/absorption is not secondary to an osmoprotec- tive effect. We propose the existence of other osmolytes or transpor- ters that function as osmoprotectors, and protect anterior hindgut Ine protein is expressed solely in the anterior hindgut. The anterior hindgut is an important site of water absorption, as demonstrated in insects other than Drosophila. In locusts, isosmotic fluid absorption SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 6 www.nature.com/scientificreports Figure 6 \| Ine in the hindgut is essential for water conservation under conditions of desiccation. (A), ine mutants are more sensitive to desiccation than WT flies. The expression of the RA or RB isoform of ine in the hindgut epithelium by hindgut-Gal4 but not by repo- or elav-Gal4 can completely rescue the sensitivity of mutants to desiccation. The median survival time (LT50) was estimated for different groups. Each bar represents the mean 6 S.E.M. Asterisk (): p , 0.01, compared with WT (t-tests, two tails). (B), survival curves demonstrate similar starvation resistance between the ine mutant and WT flies when supplied with sufficient water. The experiments were performed in triplicate (n 5 10 per genotype). The data are the means 6 S.E.M. osmotic driving force to conserve/absorb water from hindgut lumen against external hypertonicity. However, this theory lacks an explanation for how water is transferred into the hemolymph from epithelial cells, and to date, the transporter activity of ine has not been confirmed. We cannot rule out the possibility that ine may im- prove water conservation/absorption through a different, unknown mechanism. Methods Figure 6 \| Ine in the hindgut is essential for water conservation under conditions of desiccation. (A), ine mutants are more sensitive to Figure 6 \| Ine in the hindgut is essential for water conservation under conditions of desiccation. (A), ine mutants are more sensitive to desiccation than WT flies. The expression of the RA or RB isoform of ine in the hindgut epithelium by hindgut-Gal4 but not by repo- or elav-Gal4 can completely rescue the sensitivity of mutants to desiccation. The median survival time (LT50) was estimated for different groups. Each bar represents the mean 6 S.E.M. Asterisk (): p , 0.01, compared with WT (t-tests, two tails). (B), survival curves demonstrate similar starvation resistance between the ine mutant and WT flies when supplied with sufficient water. The experiments were performed in triplicate (n 5 10 per genotype). The data are the means 6 S.E.M. Fly stocks. Fly stocks were raised on standard cornmeal-agar medium with 12 hr light/12 hr dark cycles at 25uC and 60% humidity. The wild-type (WT) strain used was Canton-Special (Canton-S). The ine2 and ine3 mutants, and the transgenic flies carrying UAS-ine-RA, were kindly provided by Dr. Michael Stern18. Repo-Gal4, elav- Gal4 and UAS-GFP strains were obtained from the Drosophila Stock Center in Bloomington. The transgenic flies carrying UAS-ine-RB and hindgut-Gal4 were generated in this study (see below). hindgut-Gal4 is expressed exclusively in the hindgut epithelial cells of flies as confirmed by hindgut-Gal4 directed cytoplasmic GFP expression. desiccation than WT flies. The expression of the RA or RB isoform of ine in the hindgut epithelium by hindgut-Gal4 but not by repo- or elav-Gal4 can completely rescue the sensitivity of mutants to desiccation. The median survival time (LT50) was estimated for different groups. Each bar represents the mean 6 S.E.M. Asterisk (*): p , 0.01, compared with WT (t-tests, two tails). (B), survival curves demonstrate similar starvation resistance between the ine mutant and WT flies when supplied with sufficient water. The experiments were performed in triplicate (n 5 10 per genotype). The data are the means 6 S.E.M. Generation of hindgut-Gal4 and UAS-ine-RB transgenic flies. To generate hindgut- Gal4, we selected Irk2 (CG4370), for which predicted function implied expression in the hindgut. Fragments containing the upstream region (3R: 23518270-23521443) flanking this gene were amplified using PCR from genomic DNA (Primers: forward, 59-ATGTCAGAGCAATCAACTTCTCTTG-39; reverse, 59- TTCTCTAGACGTTTTAACTTCGCGG-39). Discussion h d In addition to the anterior hindgut, the Malpighian tubules, rec- tum, and midgut also contribute to water absorption and conser- vation in insects under conditions of external hypertonicity or desiccation. During dehydration stress, the modulation of tyramine signaling in Drosophila Malpighian tubules enhances conservation of body water41. Several anti-diuretic factors acting on the Malpighian tubules have been found. For example, CAPA-1 acts on Ncc69, the Na1-K1-2Cl2 cotransporter, to increase water absorption through an ion uptake coupled mechanism42. In addition, PKG, a cGMP- dependent kinase antagonizes the diuretic effects of tyramine and leukokinin9. The rectum can also transport water from lumen to the hemolymph33,43,44. In the locust, the chloride transport stimulating hormone (CTSH) acts to increase ion-dependent active transport of fluid from the rectum lumen45. Finally, the antidiuretic hormone RhoprCAPA-2 inhibits fluid transport into the midgut lumen in Rhodnius prolixus to conserve water13. Therefore, ine-mediated water conservation/absorption may not be the only mechanism by which systemic water homeostasis is maintained under external hypertonicity in Drosophila. Water is essential for the proper function of virtually all living cells. Organisms have developed mechanisms in the excretory system to maintain water hemostasis for a constant internal milieu under dif- ferent external osmotic conditions, such as hypertonicity. Our study reveals that hindgut expression of ine, a putative Na1/Cl2-dependent neurotransmitter/osmolyte transporter, is indispensable for the maintenance of systemic water homeostasis in Drosophila. However, further investigation of the novel mechanism mediated by ine in the hindgut is necessary to fully understand the water conservation and absorption mechanisms of Drosophila hindgut, as well as the physiological functions of the members of the Na1/ Cl2-dependent neurotransmitter/osmolyte transporter family. Methods A second gene encodes the anti-diuretic hormone in the insect, Rhodnius prolixus. Mol Cell Endocrinol 317, 53–63 (2010). Immunostaining and confocal imaging. Hindgut tissue was prepared for immunostaining as previously described48. Briefly, tissue was dissected and fixed in 100 mM glutamic acid, 25 mM KCl, 20 mM MgSO4, 4 mM sodium phosphate, 1 mM MgCl2, 4% formaldehyde for 30 min. Subsequent rinses, washes and incubations with primary and secondary antibodies were performed in 1X PBS, 0.5% BSA, 0.3% TritonX-100. Tissue was mounted in Vectashield medium (Vector Laboratories). Images were captured using confocal microscopy on an LSM 510 instrument (Zeiss). The following antibodies were used: anti-ine (15200), anti-GFP (15500), anti-b Alanine (15100) and a5 (15100). Secondary antibodies were used at 15500 and are as follows: goat anti guinea pig, goat anti-rabbit, and goat anti-mouse IgG conjugated to cy3, Alexa 488 and cy5 respectively. 13. Paluzzi, J. P., Russell, W. K., Nachman, R. J. & Orchard, I. Isolation, cloning, and expression mapping of a gene encoding an antidiuretic hormone and other CAPA-related peptides in the disease vector, Rhodnius prolixus. Endocrinology 149, 4638–4646 (2008). 14. Ianowski, J. P., Paluzzi, J. P., Te Brugge, V. A. & Orchard, I. The antidiuretic neurohormone RhoprCAPA-2 downregulates fluid transport across the anterior midgut in the blood-feeding insect Rhodnius prolixus. Am J Physiol Regul Integr Comp Physiol 298, R548–557 (2010). p y 15. Quinlan, M. C., Tublitz, N. J. & O’Donnell, M. J. Anti-diuresis in the blood-feeding insect Rhodnius prolixus Stal: the peptide CAP2b and cyclic GMP inhibit Malpighian tubule fluid secretion. J Exp Biol 200, 2363–2367 (1997). 16. Wiehart, U. I., Nicolson, S. W., Eigenheer, R. A. & Schooley, D. A. Antagonistic control of fluid secretion by the Malpighian tubules of Tenebrio molitor: effects of diuretic and antidiuretic peptides and their second messengers. J Exp Biol 205, 493–501 (2002). Viability assays on hypertonic media. Flies were collected for 4 days following eclosion. Instant fly food medium (Carolina) was prepared according to the manufacturer’s instructions. Hypertonic medium was prepared by replacing water with 0.2 M NaCl or KCl solution. Adult flies of the indicated genotype (10 per vial) were maintained on either normal or hypertonic medium for 10 days. Live and dead flies were counted daily. Fly manipulations and assays were conducted at room temperature and ambient humidity18. 17. Eigenheer, R. A., Nicolson, S. W., Schegg, K. M., Hull, J. J. & Schooley, D. A. Methods Identification of a potent antidiuretic factor acting on beetle Malpighian tubules. Proc Natl Acad Sci U S A 99, 84–89 (2002). 18. Huang, X. et al. The Drosophila inebriated-encoded neurotransmitter/osmolyte transporter: dual roles in the control of neuronal excitability and the osmotic stress response. Genetics 160, 561–569 (2002). Trypan blue staining. WT and ine3 flies were maintained on either normal or hypertonic medium for 4 days. Hindgut tissue was prepared for Trypan blue staining as previously described32. Briefly, tissue was dissected in 1X PBS, immersed in 0.2 mg/ ml Trypan Blue in 1X PBS, and rotated for 30 min at room temperature. After washing in PBS for 30 min, the tissue was immediately scored for Trypan Blue staining of the anterior hindgut. Scoring was based on an index of the anterior hindgut: no color, 0; any blue, 1; darkly stained nuclei, 2; large patches of darkly stained cells, 3; or complete staining of most cells in the tissue, 4. 19. Soehnge, H. et al. A neurotransmitter transporter encoded by the Drosophila inebriated gene. Proc Natl Acad Sci U S A 93, 13262–13267 (1996). 20. Amara, S. G. & Kuhar, M. J. Neurotransmitter transporters: recent progress. Annu Rev Neurosci 16, 73–93 (1993). 21. Borden, L. A., Smith, K. E., Hartig, P. R., Branchek, T. A. & Weinshank, R. L. Molecular heterogeneity of the gamma-aminobutyric acid (GABA) transport system. Cloning of two novel high affinity GABA transporters from rat brain. J Biol Chem 267, 21098–21104 (1992). 22. Rasola, A., Galietta, L. J., Barone, V., Romeo, G. & Bagnasco, S. Molecular cloning and functional characterization of a GABA/betaine transporter from human kidney. FEBS LETT 373, 229–233 (1995). Hemolymph volume and body water measurement. Hemolymph volume and body water were estimated as previously described49. Flies of the indicated genotype were maintained on normal or hypertonic media for 4 days. Adult flies from each genotype were anesthetized with CO2 and weighed. The abdomen of each fly was gently torn and hemolymph was blotted from the abdominal opening with a Kimwipe that had been slightly moistened with isotonic saline. Each fly was then weighed a second time, then dried for 1 h at 60uC and weighed a third time. Hemolymph volume was estimated by determining the reduction in mass following hemolymph blotting. Percentage of total body water and hemolymph were estimated. 23. Chiu, C., Ross, L. S., Cohen, B. N., Lester, H. A. & Gill, S. Methods S. The transporter-like protein inebriated mediates hyperosmotic stimuli through intracellular signaling. J Exp Biol 203, 3531–3546 (2000). 24. Kempson, S., Vovor-Dassu, K. & Day, C. Betaine transport in kidney and liver: Use of betaine in liver injury. Cell Physiol Biochem 32, 32–40 (2013). 25. Burg, M. B. & Ferraris, J. D. Intracellular organic osmolytes: function and regulation. J Biol Chem 283, 7309-7313 (2008). 26. Zhou, Y. et al. The betaine-GABA transporter (BGT1, slc6a12) is predominantly expressed in the liver and at lower levels in the kidneys and at the brain surface. Am J Physiol-Renal 302, F316–F328 (2012). Desiccation resistance and starvation resistance. To evaluate desiccation resistance, 3-day-old male flies were placed in empty glass shell vials (10 flies per vial) and introduced into a Plexiglas desiccation chamber. The temperature was maintained at 24–25uC. The number of dead flies was scored at an hourly interval until all of the flies had died. For starvation resistance, 3-day-old male flies were introduced into vials containing 10 mL of 0.5% agar in groups of 10 flies per vial. The vials were changed to fresh medium every 48 h. Deaths were scored three times per day until all of the flies had died. Each genotype was tested three times50. y 27. Kempson, S. A. & Montrose, M. H. Osmotic regulation of renal betaine transport: transcription and beyond. Pflu¨gers Archiv 449, 227–234 (2004). 28. Fox, D. T. & Spradling, A. C. The Drosophila hindgut lacks constitutively active adult stem cells but proliferates in response to tissue damage. Cell Stem Cell 5, 290–297 (2009). 29. Chintapalli, V. R., Wang, J. & Dow, J. A. Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39, 715–720, doi:10.1038/ng2049 (2007). Statistical analysis. Statistical significance was determined using an unpaired Student’s t-test (two-tailed). P-values of less than 0.01 were considered significant. 30. 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J Exp Biol 206, 1183–1192 (2003). 7. Aikins, M. J. et al. Vasopressin-like peptide and its receptor function in an indirect diuretic signaling pathway in the red flour beetle. Insect Biochem Mol Biol 38, 740–748 (2008). 37. Wessing, A. & Eichelberg, D. in The Genetics and Biology of Drosophila (Ashburner, A. & Wright, T. R. F.) 1–42 (Academic Press, London, 1978). 8. Spring, J. H., Morgan, A. M. & Hazelton, S. R. A novel target for antidiuretic hormone in insects. Science 241, 1096–1098 (1988). 38. Stergiopoulos, K., Cabrero, P., Davies, S.-A. & Dow, J. A. Salty dog, an SLC5 symporter, modulates Drosophila response to salt stress. Physiol Genomics 37, 1 (2009). 38. Stergiopoulos, K., Cabrero, P., Davies, S.-A. & Dow, J. A. Salty dog, an SLC5 d l h l l h l 9. Ruka, K. A., Miller, A. P. & Blumenthal, E. M. Inhibition of diuretic stimulation of an insect secretory epithelium by a cGMP-dependent protein kinase. Am J Physiol Renal Physiol 304, F1210–1216 (2013). symporter, modulates Drosophila response to salt stress. Physiol Genomics 37, 1 (2009). Methods The fragments were cloned, sequence- verified, and inserted into the pG4PN vector upstream of the Gal4 cassette. The final DNA construct was injected into w1118 embryos by BestGene, Inc. (Chino Hills, CA)46. We analyzed the expression pattern of hindgut-Gal4 by driving expression of a UAS-GFP reporter gene, and detecting fluorescence using confocal microscopy of whole mount tissue (Fig. 1). Wild-type ine-RB cDNA was obtained by RT-PCR (Primers: forward, 59- ATGCCGAACCGCCAGGACTACGAT-39; reverse, 59- CTACTGGCCACATGGTCCTCCTGCC-39), subcloned into a pUAST vector, and injected into w1118 flies by BestGene, Inc. (Chino Hills, CA) to generate UAS-ine-RB transgenic flies. in the anterior hindgut is driven by an apical membrane electrogenic Cl2 pump. The antidiuretic hormone Schgr-ITP acts on the locust hindgut via cyclic AMP and GMP to increase the conductance of both K1 and Na1 and to stimulate the Cl2 pump. As a result of the increased ion uptake, water absorption increases39,40. It remains unknown, however, whether similar ion-uptake-coupled water absorption mechanisms are present in the Drosophila hindgut. We found that loss of ine in the anterior hindgut epithelium causes severe dehydration in response to a hypertonic diet, and higher rates of body water loss under desiccation, which suggests the existence of a new mechanism of water conservation/absorption in the hindgut of Drosophila mediated by ine. We propose above that ine transports osmolytes across the plasma membrane from the hemolymph and accumulates osmolytes within the hindgut epithelium, generating an Antibodies. The ine antibody was raised in guinea pig against a GST-fused fragment of ine protein (C-terminal portion of ine, 847-943a.a.). The antibody was affinity purified by coupling the antigen to Sepharose 4B. The specificity of the antibody was validated by immunostaining of the null mutant ine3. Rabbit polyclonal Anti-GFP antibody was purchased from Life Technologies. Rabbit polyclonal anti-b alanine antibody (ab37076), purchased from Abcam (Cambridge, MA) was used to label the SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 7 www.nature.com/scientificreports www.nature.com/scientificreports general gut structure of adult Drosophila. Mouse monoclonal antibody a5-IgG, specific for the a-subunit of the Na1/K1-ATPase, was obtained from The University of Iowa Developmental Studies Hybridoma Bank47. All secondary antibodies were purchased from Jackson ImmunoResearch. 11. Paluzzi, J. P., Park, Y., Nachman, R. J. & Orchard, I. Isolation, expression analysis, and functional characterization of the first antidiuretic hormone receptor in insects. Proc Natl Acad Sci U S A 107, 10290–10295 (2010). 12. Paluzzi, J. P. & Orchard, I. Author contributions 43. Phillips, J. E. Rectal Absorption in the Desert Locust, Schistocerca Gregaria Forskal. Iii. The Nature of the Excretory Process. J Exp Biol 41, 69–80 (1964). 43. Phillips, J. E. Rectal Absorption in the Desert Locust, Schistocerca Gregaria Forskal. Iii. The Nature of the Excretory Process. J Exp Biol 41, 69–80 (1964). Z.L. and H.L. designed the experiments; Z.L. performed the experiments; Z.L., H.L. and C.Q. wrote the manuscript. All authors reviewed the manuscript. Z.L. and H.L. designed the experiments; Z.L. performed the experiments; Z.L., H.L. and C.Q. wrote the manuscript. All authors reviewed the manuscript. y p 44. Bradley, T. J. & Phillips, J. E. The secretion of hyperosmotic fluid by the rectum of a saline-water mosquito larva, Aedes taeniorhynchus. J Exp Biol 63, 331–342 (1975). q y p 45. Proux, B., Proux, J. & Phillips, J. Antidiuretic action of a corpus cardiacum factor (CTSH) on long-term fluid absorption across locust recta in vitro. J Exp Biol 113, 409–421 (1984). Methods 39. Audsley, N., Jensen, D. & Schooley, D. A. Signal transduction for Schistocerca gregaria ion transport peptide is mediated via both cyclic AMP and cyclic GMP. Peptides 41, 74–80 (2013). y 10. Coast, G. M., Orchard, I., Phillips, J. E. & Schooley, D. A. Insect diuretic and antidiuretic hormones. Adv Inesct Physiol 29, 279–409 (2002). SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 8 www.nature.com/scientificreports Acknowledgments 40. Audsley, N. & Phillips, J. E. Stimulants of ileal salt transport in neuroendocrine system of the desert locust. Gen Comp Endocrinol 80, 127–137 (1990). g We thank Bloomington Drosophila Stock Center and Developmental Studies Hybridoma Bank for fly stocks and antibodies. This work is supported by NIH grants R01AG022508 and R01EY021796 awarded to H.-S. L. y p 41. Blumenthal, E. M. Modulation of tyramine signaling by osmolality in an insect secretory epithelium. Am J Physiol Cell Physiol 289, C1261–1267 (2005). 42. Rodan, A. R., Baum, M. & Huang, C. L. The Drosophila NKCC Ncc69 is required for normal renal tubule function. Am J Physiol Cell Physiol 303, C883–894 (2012). Additional information Competing financial interests: The authors declare no competing financial interests. Competing financial interests: The authors declare no comp Competing financial interests: The autho How to cite this article: Luan, Z., Quigley, C. & Li, H.-S. The putative Na1/Cl2-dependent neurotransmitter/osmolyte transporter inebriated in the Drosophila hindgut is essential for the maintenance of systemic water homeostasis. Sci. Rep. 5, 7993; DOI:10.1038/srep07993 (2015). 46. Miller, C. J. & Carlson, J. R. Regulation of odor receptor genes in trichoid sensilla of the Drosophila antenna. Genetics 186, 79–95 (2010). 47. Baumann, O., Salvaterra, P. M. & Takeyasu, K. Developmental changes in b- subunit composition of Na, K-ATPase in the Drosophila eye. Cell Tissue Res 340, 215–228 (2010). This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permissionfrom the licenseholderin order toreproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ 48. Ohlstein, B. & Spradling, A. Multipotent Drosophila intestinal stem cells specify daughter cell fates by differential notch signaling. Science 315, 988–992 (2007). 49. Folk, D. G., Han, C. & Bradley, T. J. Water acquisition and partitioning in Drosophila melanogaster: effects of selection for desiccation-resistance. J Exp Biol 204, 3323–3331 (2001). 50. Matzkin, L. M., Watts, T. D. & Markow, T. A. Evolution of stress resistance in Drosophila: interspecific variation in tolerance to desiccation and starvation. Funct Ecol 23, 521–527 (2009). SCIENTIFIC REPORTS \| 5 : 7993 \| DOI: 10.1038/srep07993 9	14,767
https://openalex.org/W4299862309	OpenAlex	Open Science	CC-By	2,016	Kuba a saját útját járja	Gabòr Kovàcs	Hungarian	Spoken	678	2,017	Kuba a saját útját járja Kuba a saját útját járja 2022. 07. 08. 0:44 2022. 07. 08. 0:44 https://www.magyardiplo.hu/archivum/2016/211-2016-junius/2212-kuba-a-sajat-utjat-jarja.html?tmpl=component&print= Kuba a saját útját járja Kategória: 2016. június Írta: Kovács Gábor Kategória: 2016. június Írta: Kovács Gábor Julio César Cancio Ferrer, a Kubai Köztársaság nagykövete fogadta lapunk munkatársát és válaszolt kérdéseire. A szigetországban szeptember 20-ig tartó nemzeti konzultációval kapcsolatban a Nagykövet elmondta, hogy a Kubai Kommunista Párt VII. Kongresszusa határozatának megfelelően országos vita zajlik a gazdasági és társadalmi modell aktualizálásáról és a 2030-ig tartó fejlesztési tervről, amelynek alapvető célja a társadalom szocialista jellegének az erősítése. Hangsúlyozta, hogy szakítani kell a régi beidegződésekkel és alkalmazkodni a kialakult helyzethez. Csökkenteni kívánják a bürokráciát, és a döntéseket alsóbb szintre akarják helyezni. Megjegyezte, hogy foglalkoznak az 1992- ben elfogadott alkotmány módosításával is. A jelenlegi helyzetben kiemelt kérdés a gazdaság hatékonyságának a növelése. A kubai vezetés elismeri a magántulajdont, ami a forradalom után elsősorban a mezőgazdaságban alakult ki. A szövetkezeti ágazatban is jelentős változások történtek, amit a magántulajdon egyik formájának tekintenek. Az elmúlt években a szövetkezetek tevékenysége a lakosság igényeit kielégítő szolgáltatásokra is kiterjedt. A Nagykövet kijelentette, hogy fokozott figyelmet fordítanak a piaci hatások figyelembe vételére, de az országban nincs úgynevezett szabad piac. A munkavállalók mintegy 20 százaléka a magánszektorban dolgozik, a gazdaság alapja azonban a társadalmi tulajdon. A magánvállalkozások a jövőben sem léphetnek közvetlen üzleti kapcsolatba külföldi cégekkel vagy bankokkal. A kettős pénzrendszer megszűntetésével kapcsolatban Julio César Cancio Ferrer kijelentette, hogy ez egy hosszú és bonyolult folyamat, amit nagy óvatossággal kell kezelni. A kormányzat el akarja kerülni, hogy a pénzrendszer megváltoztatása lehetőséget adjon neoliberális jellegű megszorításokra. A nemzetközi pénzügyi kapcsolatokról szólva, a Nagykövet elmondta, hogy az adósságokat Oroszországgal, Kínával, a nyugati országokat tömörítő Párizsi Klub tagjaival és más országokkal már újratárgyalták. A megváltozott helyzetet figyelembe vevő, új megállapodásokból adódó kötelezettségeket Kuba pontosan be fogja tartani. (Oroszország 2013 decemberében leírta a szovjet idökből származó 32 milliárd dolláros adósság 90 százalékát. A Párizsi Klub államai a 11 milliárd dolláros kubai adósságból 8,5 milliárdot elengedtek. A szerkesztő). Barack Obama amerikai elnök havannai látogatása óta a két ország kapcsolataiban történt változásokról a diplomata kijelentette, hogy előre lépés történt járványügyi és egészségügyi kérdésekben valamint a sport kapcsolatokban. Szakértők megállapodtak a légiforgalom bővítéséről is. Az elnök kezdeményezte, hogy a Kongresszus oldja fel a Kuba elleni embargót, de ez változatlanul érvényben van. Az USA nem hajlandó kiüríteni a Kubához tartozó Guantánamo katonai bázist sem. A 1/2 ps://www.magyardiplo.hu/archivum/2016/211-2016-junius/2212-kuba-a-sajat-utjat-jarja.html?tmpl=component&print=1 2022. 07. 08. 0:44 Kuba a saját útját járja Kuba a saját útját járja kétoldalú viszonyt nehezíti, hogy az Egyesült Államok kormánya folyamatosan a rendszerellenes csoportokat támogat Kubában. https://www.magyardiplo.hu/archivum/2016/211-2016-junius/2212-kuba-a-sajat-utjat-jarja.html?tmpl=component&print= Kategória: 2016. június Írta: Kovács Gábor kétoldalú viszonyt nehezíti, hogy az Egyesült Államok kormánya folyamatosan a rendszerellenes csoportokat támogat Kubában. Nemzetközi kérdésekre áttérve, a Nagykövet hangsúlyozta, hogy az USA offenzívát indított Latin-Amerikában. Washington szövetségesének tekinti a térség nemzeti oligarcháit. A közvélemény manipulálásával megpróbálják egyes országokban befolyásolni a választások eredményét. A neoliberális tőkével összefonódott média támadja a baloldali kormányokat. A haladó erők és a reakciós csoportok között éles küzdelem folyik Venezuelában, Brazíliában, Bolíviában és más országokban. Latin-Amerikában egy második függetlenségi harc zajlik, ami a kubai forradalom győzelmével kezdődött. Az elmúlt évtizedekben az USA mindent megtett, hogy megbuktassa Fidel Castrot, de Obama elnök végül elismerte az elszigetelési politika kudarcát. A nyugati média gyakran hasonlítja a kubai „kiigazítást” a Vietnamban történő változásokhoz. Kérdésemre Julio César Cancio Ferrer kijelentette, hogy mások vagyunk és a saját utunkat járjuk. Kubában nincs szó gazdasági pluralizmusról és a magánszektor szerepe átmeneti jellegű. Meg kell érteni, hogy 90 mérföldre az USA partjaitól az egypártrendszer a szocialista átalakulás folytatásának a biztosítéka. A kubai-magyar kapcsolatokat Julio César Cancio Ferrer pragmatikusnak nevezte. A két ország között nincsenek vitás kérdések. Kijelentette, hogy a kölcsönös előnyök és a belügyekbe való be nem avatkozás elve alapján készek az együttműködés fejlesztésére a kereskedelemben, a kultúra valamint a sport területén. Szeretnék bővíteni a turista forgalmat és napirenden van közös vállalatok létesítése. Az együttműködésnek nagy hagyományai vannak, mert a rendszerváltásig több ezer magyar szakember dolgozott a szigetországban és sok kubai diák tanult a magyar egyetemeken. A korábbi kapcsolatokat és a közös tapasztalatokat azonban jobban ki kellene használni. nemzetközi kapcsolatok kuba szocializmus 2/2	19,143
W2972421431.txt_1	Open-Science-Pile	Open Science	Various open science	2,020	Experimental long-term effects of early-childhood and school-age exposure to a conditional cash transfer program	None	English	Spoken	7,017	9,900	Experimental long-term effects of early-childhood and school-age exposure to a conditional cash transfer program Teresa Molina Millan, Karen Macours, John Maluccio, Luis Tejerina To cite this version: Teresa Molina Millan, Karen Macours, John Maluccio, Luis Tejerina. Experimental long-term effects of early-childhood and school-age exposure to a conditional cash transfer program. Journal of Development Economics, 2020, 143, �10.1016/j.jdeveco.2019.102385�. �halshs-02297704� HAL Id: halshs-02297704 https://shs.hal.science/halshs-02297704 Submitted on 20 Jul 2022 HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Distributed under a Creative Commons Attribution 4.0 International License Version of Record: https://www.sciencedirect.com/science/article/pii/S0304387818312768 Manuscript_08127d4a92f6a678d2e5f37459a6651a Experimental Long-Term Effects of Early-Childhood and SchoolAge Exposure to a Conditional Cash Transfer Program Teresa Molina Millán, Karen Macours, John A. Maluccio, and Luis Tejerina1 This version: June 2019 Abstract Numerous evaluations of conditional cash transfer (CCT) programs show positive short-term impacts, but there is only limited evidence on whether these benefits translate into sustained longer-term gains. This paper uses the municipal-level randomized assignment of a CCT program implemented for five years in Honduras to estimate long-term effects 13 years after the program began. We estimate intent-to-treat effects using individual-level data from the population census, which allows assignment of individuals to their municipality of birth, thereby circumventing migration selection concerns. For the non-indigenous, we find positive and robust impacts on educational outcomes for cohorts of a very wide age range. These include increases of more than 50 percent for secondary school completion rates and the probability of reaching university studies for those exposed at school-going ages. They also include substantive gains for grades attained and current enrollment for others exposed during early childhood, raising the possibility of further gains going forward. Educational gains are, however, more limited for the indigenous. Finally, exposure to the CCT increased the probability of international migration for young men, from 3 to 7 percentage points, also stronger for the non-indigenous. Both early childhood exposure to the nutrition and health components of the CCT as well as exposure during school-going ages to the educational components led to sustained increases in human capital. JEL Classification: I25, I28, I38, O15 Keywords: conditional cash transfers (CCTs), early childhood, education, migration 1 Molina Millán is at Nova School of Business and Economics (teresa.molina@novasbe.pt), Macours at Paris School of Economics and INRA (karen.macours@psemail.edu), Maluccio at Middlebury College (maluccio@middlebury.edu), and Tejerina at the Inter-American Development Bank (IDB) (luist@iadb.org). This research was supported by funds from the IDB Economic and Sector Work program “CCT Operational Cycles and Long-Term Impacts” (RG-K1422) and the French National Research Agency (ANR) under grant ANR-17-EURE0001. We thank the government of Honduras and the National Institute of Statistics for permission to work with the census microdata. We also thank Tania Barham, Pablo Ibarrarán, Norbert Schady, Marco Stampini, Guy Stecklov, two anonymous referees, and participants in presentations at the IDB, Middlebury, NEUDC 2018 and Vassar for valuable comments and suggestions. All remaining errors are our own. The content and findings of this paper reflect the opinions of the authors and not those of the IDB, its Board of Directors, or the countries they represent. © 2019 published by Elsevier. This manuscript is made available under the CC BY NC user license https://creativecommons.org/licenses/by-nc/4.0/ 1. Introduction Conditional cash transfer (CCT) programs are among the most popular social programs in the developing world. They have been operating in Latin America for two decades, reaching 25 percent of the region’s population (Robles, Rubio and Stampini, 2017), and increasingly in other regions. CCTs aim to alleviate short-run poverty while inducing investment in the nutrition, health, and education of the next generation. Evidence from various contexts demonstrates their effectiveness in the short run (Fiszbein and Schady, 2009).2 There is greater uncertainty, however, as to whether these translate into longer-term gains (Molina Millán et al., 2019). More generally, little is known about whether and how CCTs affect the trajectories of children benefitting directly or indirectly from different program components at different points during their childhood. This paper provides experimental evidence on long-term impacts for children exposed at different stages of early childhood and school-ages to five years (2000–2005) of a Honduran CCT program, the Programa de Asignación Familiar (PRAF-II). This CCT, similar in design to other programs in the region, provides a unique opportunity to study long-term impacts because it was randomized across 70 municipalities and, unlike most other randomized CCT evaluations such as Mexico’s PROGRESA, control municipalities were never phased into the program. Exploiting the municipality-level randomized assignment, we use individual-level data from the national census, collected 13 years after the program began (and thus eight years after it ended), to analyze impacts of the CCT on individuals in cohorts spanning nearly 25 years. Assigning each individual to the municipality where they were born—a good proxy for their preprogram location—we circumvent the typical selection and attrition concerns that affect the study of long-term impacts of highly mobile cohorts of individuals. We thus estimate intent-totreat (ITT) impacts that account for migration within the national territory, as high as 30 percent for some cohorts of interest. The census also includes information on current international migration of former household members, allowing direct study of international migration as an outcome and assessment of such migration as another potentially important selection concern.3 Beyond these key advantages, the census data provide sufficient statistical power for the estimation of long-term impacts on several different cohorts of interest, and we can separately 2 Recent evidence on short-term effects of CCTs on education includes reviews (Murnane and Ganimian, 2014; Glewwe and Muralidharan 2015) and meta-analyses (Saavedra and García, 2012; Baird et al. 2014; McEwan 2015). 3 An important limitation of using census data, however, is that we are unable to capture measures of nutrition or health improvements. 1 estimate the impacts on children exposed to the nutrition and health components of the program during early childhood and to the education components of the program at older school-going ages, as well as estimate the impacts on those who benefitted (partially) from both. We also analyze whether there are spillover or indirect effects on others by examining cohorts of children who were too old to have been directly affected by the education conditionalities when the program started in 2000 as well as children born after the program ended. The wide age range examined in a single setting constitutes an important contribution of this paper, providing a better understanding on whether exposure to CCTs at different ages can impact human capital and subsequent outcomes. This is particularly relevant as some transfer programs narrowly target specific ages. While the first generation of CCT programs in Latin America typically covered a wide age range, as was the case in Honduras, more recent programs in Asia (Filmer and Schady, 2014; Levere, Acharhya and Bharadwaj, 2016) and Africa (Baird, McIntosh and Özler, 2011; Benhassine et al, 2015) often target narrower populations and objectives (e.g., only nutrition and health in very early childhood; only educational outcomes at critical ages in primary or secondary school). Beyond the cash transfer literature, examination of impacts at different ages of exposure is relevant for the literature on human capital formation. Indeed, economists often motivate focusing on early childhood based on Cunha and Heckman’s (2007) multistage model of skill formation that predicts “skill begets skill” with investments made in early life being favored over those made later in childhood. Whether investments in early life translate into outcomes during a later phase in childhood of course also depends on how well each component of the CCT was implemented, exposure to other programs that may differ by cohort, and any remaining constraints that beneficiaries may face as they grow up. As no identifiable exogenous variation in program implementation or later program exposure exists, we abstract from such effects in this paper, as is done in most studies analyzing long-term impacts. However, the census data do provide sufficient power to study program impacts separately across groups likely to face different constraints in both the shortand long-term in the Honduran context, and thus provide insights on the potential importance of other constraints. Specifically, we analyze impacts by sex and ethnic background (nonindigenous versus indigenous), resulting in four distinct demographic groups. Because no other variables in the census can reasonably be considered exogenous 13 years after the start of the program, these are the only four groups for which we can examine heterogeneity at the individual level without introducing the typical concerns regarding the use of specification searches in heterogeneity analysis. The groups provide important variation in possible 2 constraints. As in many low- and middle-income countries, education and labor market decisions for men and women are quite different, with women experiencing much lower labor market participation and stronger interactions between labor market and fertility outcomes than men. And similar to those in many other Latin American countries, the indigenous population in Honduras has long suffered from higher poverty, poorer access to markets, and labor market discrimination, which together with a strong emphasis on community ties and attachment to the land may make them less mobile (World Bank, 2006; UNSR, 2016). Despite the vast literature on CCT programs, quantitative work specifically examining program impacts on indigenous populations remains relatively scarce. There is a body of ethnographic work that points to specific challenges related to CCT and related programming for indigenous populations (Correa Aste and Roopnaraine, 2014). For many programs in Latin America, there have been efforts to improve targeting to indigenous populations, but less has been done to adapt programs to better accommodate indigenous cultures or the particular challenges they face. Programs targeting the nuclear family, for example, may not adequately reach the person or persons in charge of making decisions about education and health spending. PRAF-II, to our knowledge, took no explicit measures specifically related to indigenous beneficiaries (Hernandez Ávila, 2011). Finally, another key advantage of using individual census data is that it allows reliable estimation of impacts on rare outcomes. This is relevant for international migration, which is only 1–3 percent for the oldest cohorts in the control group. It is even more salient for the highest levels of education. Less than 1 percent of the older cohorts, for example, have some universitylevel education. Very early teenage pregnancy is another key outcome variable that can be analyzed for the same reason. All of these are important for understanding the potential longterm impacts of CCTs. And for some such outcomes, even the short-term evidence is relatively scarce and inconclusive for similar reasons. For international migration, in particular, two studies with experimental estimates of the short-term impacts of the Mexican CCT program show opposite results (Stecklov et al., 2005; Angelucci, 2015). Given the wide reach of CCT programs, evidence on migration impacts can not only help understand potential selection biases but also inform the more general international migration policy debate. We find that the Honduran CCT led to long-term significant increases in schooling for both women and men, including at the university level, well beyond the primary school grades directly targeted by the program. They also include substantive gains for grades attained and current enrollment for children exposed during early childhood, raising the possibility of further 3 gains going forward. Effects for indigenous beneficiaries, however, are much more limited than those for the non-indigenous. We also find significant effects on international migration (though from a small base), a result that demonstrates how program exposure can set children on different pathways and have potentially important public policy implications. As such, this paper complements other recent evidence on long-term impacts of CCTs (see Molina Millán et al. (2019) for a review). It is closely related to long-term impact studies exploiting the randomized phase-in of cash transfer interventions in Mexico (Behrman, Parker and Todd, 2009, 2011; Fernald, Gertler and Neufeld, 2009), Nicaragua (Barham, Macours and Maluccio, 2013, 2019) and Ecuador (Araujo, Bosch and Schady, 2018). There are also clear parallels with Parker and Vogl (2018), who use Mexican census data and the non-experimental national rollout of PROGRESA to analyze differential long-term impacts. This paper differs from those studies in its ability to experimentally estimate absolute long-term impacts. Other studies estimating absolute long-term impacts include: 1) Barrera-Osorio, Linden, and Saavedra (2019), who study impacts 13 years after an individually randomized educational CCT in urban Colombia using administrative data for a specific cohort targeted by the intervention; 2) Baird, McIntosh, and Özler (2018), who also study impacts of an educational CCT in Malawi two years after it ended; and 3) Cahyadi et al. (2018), who study the six-year absolute impacts of an ongoing Indonesian CCT program on ages ranging from 0 to 15 at the start of the program.4 For children exposed during school-going ages, the existing evidence from several of the above studies generally indicates that CCTs help them obtain higher education, but is less conclusive for subsequent outcomes. Evidence on relatively rare events, such as international migration or university studies, is also limited. The evidence base is even narrower for children exposed to the nutrition and health components of CCTs during early childhood, with several experimental differential studies suggesting fadeout of impacts or catch-up of original control groups that received similar benefits a little bit later in life, while other studies point to positive long-term effects on cognition and education. Most estimates are, however, for programs that are ongoing, and it is often not possible to disentangle whether the estimated impacts are driven by the cumulative exposure to the CCT since early childhood or are instead capturing short-term impacts of the start of the schooling conditionality and transfers when children reach school age. Given that the program we study had ended prior to the census data collection, this paper can isolate the long-term impacts of early childhood exposure alone. Overall, we contribute to the 4 More broadly, this paper also relates to the mixed emerging longer-term evidence for unconditional cash transfers (Bandiera et al., 2017; Banerjee et al., 2016; Handa et al., 2018; Haushofer and Shapiro, 2018). 4 CCT literature by providing experimental evidence on the absolute long-term impacts of program exposure during a wide range of ages in early childhood and adolescence, and for a program that ended eight years earlier. 2. The Honduran CCT Program and Prior Evidence We study the long-term impacts of the second phase of PRAF, a CCT implemented from 2000 to 2005. PRAF-II aimed to increase investment in human capital, including nutrition and health during early childhood and education during primary-school ages. Modeled after the PROGRESA program in Mexico, PRAF-II provided cash transfers (in the form of readily exchangeable vouchers) to: 1) households with pregnant women and (initially) children ages 0– 3 (extended to age five in 2003), conditional on attendance at prenatal and child health and growth monitoring appointments and health education workshops; and 2) households with children ages 6–12 who had not yet completed fourth grade, conditional on school enrollment and attendance. Transfers averaged 4 percent of total preprogram household income, relatively little compared to other CCTs in the region, and were delivered twice annually. In randomly selected areas, the program also aimed to strengthen the supply-side through investments in the quality of both health and education services (IDB, 1998, 2006; IFPRI, 2003; Moore, 2008).5 The CCT targeted 70 rural municipalities in western Honduras with among the highest malnutrition rates in the country, and a municipality-level randomized assignment was used to determine treatment and control municipalities for evaluation. Randomization was stratified into five blocks of 14 municipalities each, after ordering them by malnutrition levels (Morris et al., 2004). In the randomly selected treatment municipalities, all households with children in the specified age groups were eligible to receive program benefits for up to five years until 2005, after which the program ended. The control municipalities never received the program, an essential feature that allows estimation of the absolute long-term program impacts by comparing outcomes of children born in experimental treatment versus control municipalities (IDB, 2006). The evaluation design included three different benefit packages and a control group: 1. G1: Households received cash transfers conditional on nutrition, health, and education behaviors (20 municipalities). We refer to this as the “basic CCT.” 5 Appendix B provides further information about the CCT, as well as other related interventions implemented in program municipalities in later years. We treat all other interventions post-randomization as potentially endogenous and therefore do not control for them in the main analyses. 5 2. G2: Households received cash transfers conditional on nutrition, health, and education behaviors, and schools and health centers received direct investments and support (20 municipalities). 3. G3: Schools and health centers received direct investments and support, but households did not receive any direct benefits (10 municipalities). 4. G4: The control group (20 municipalities). Program monitoring documents and short-term evaluation reports indicate that the health and schooling supply-side interventions in G2 and G3 were implemented with considerable delays and not fully operational until after 2002. The reports do not allow a full characterization of these delays, which may have disrupted health and education services and/or affected in unknown ways individual perceptions and expectations in G2 and G3. Therefore, while we adhere to the experimental design accounting for all three treatment arms, we focus our discussion on the impacts of the basic CCT components (G1).6 Emphasis on the basic CCT has the additional advantage of making the analysis more comparable to most existing research on the long-term impacts of CCTs. Prior evidence from short-term evaluations shows impacts on early-life health indicators and schooling that are qualitatively similar to those found for other CCTs in the region. Morris et al. (2004) examine the short-term program evaluation data and find that after two years the basic CCT increased the uptake of prenatal care (five or more visits) for pregnant women by nearly 20 percentage points (on a base of about 50 percent). They find similarly large increases for routine checkups (including growth monitoring) for children under 3 years, which they suggest could be important for administering vaccinations more opportunely, though there was little evidence of improved vaccination at that stage. Effects on schooling were more modest than observed in other contexts, however, possibly reflecting the relatively small transfer size in comparison to other programs in the region. Galiani and McEwan (2013) use the 2001 national census, administered after eight months of transfers, and find an increase in enrollment rates of about 8 percentage points among children eligible for the educational transfer and a decrease of 3 percentage points in the probability of having worked in the last week, with larger effects in the two strata with the highest levels of malnutrition at baseline. Glewwe and Olinto (2004) use the short-term program evaluation data after two years and show significant but smaller increases in primary school enrollment rates of around 3 percentage points for children 6–12 years old at 6 Estimates for G2 are presented in Appendix D. 6 baseline as well as modest improvements in attendance and grade promotion, and a slight reduction in hours worked. In other prior work, Stecklov et al. (2007) demonstrate that the program led to an increase in fertility of 2–4 percentage points by 2002. These changes in fertility point to the possibility that cohorts born after the start of the program in 2000, the younger cohorts we analyze, could be selective.7 In addition, if such changes in fertility in turn led to a shift in fertility norms, they also may have had indirect effects on the older cohorts reaching reproductive ages. Lastly, examining the same period as we do, Ham and Michelson (2018) use municipalitylevel averages from both the 2001 and 2013 Honduran censuses to analyze the impact of PRAF-II for children ages 6–12 in 2001 (and thus 18-24 in 2013). They exploit the randomized design and estimate municipal level differences-in-differences for this age cohort, showing increases in grades attained, secondary school completion, and labor force participation, especially for females in G2, after controlling for municipality-level fixed effects and a number of time-variant (and hence possibly endogenous) controls. As the analysis uses average outcomes based on place of residence in 2001 and 2013, it makes the strong assumption that migration between 2001 and 2013 (over 25 percent for thus age cohort) does not affect the internal validity and the estimates do not account for any returns that materialize through migration. 3. Data and Methodology The principal data source is the XVII Honduran National Population and Housing Census of 2013.8 For complementary analyses, we also use the 1988 and 2001 national censuses, and the short-term program evaluation data collected for PRAF-II. For the main analyses in 2013, we limit the census sample to all individuals born in the 70 targeted rural municipalities regardless of their current residential location. For the age cohorts we study, municipality of birth together with the municipal-level randomized program assignment provides an exogenous indicator of program exposure not influenced by subsequent domestic migration or geographical sorting that may have occurred during or after the program.9 In addition to including other typical 7 Program rules were altered in 2003, removing this possible fertility incentive, so that the short-term fertility increase most likely affected only those born in the first years after the start of the program, i.e. those ages 9–12 in 2013. 8 Below we consider in more detail potential problems with national census data; for a general description, see Cleland (1996). 9 We separately examine the likelihood that different age cohorts born before the program were still living in their municipality of birth at the time the program started (section 4.4); in general such preprogram migration would attenuate estimates relative to ones for which residential location at the moment of program assignment was used. 7 information (sex, age, ethnicity, education, migration, civil status, and fertility) for all current residents, the census includes basic information on former household members who left Honduras at any point over the prior decade. Information available on these international migrants includes sex, age, year of migration, and current country of residence—but not schooling. To estimate program impacts on international migration itself, we incorporate the migrants into the individual-level census sample by assuming they were born in the municipality where the household from which they migrated is located in 2013.10 An implication of targeting areas with the highest malnutrition rates in the country was that PRAF-II operated in regions with a high share of indigenous people. While the indigenous in Honduras comprise only 6.5 percent of the national population, they make up 39 percent of the main analysis sample (individuals ages 6–29 in 2013 and born in one of the 70 program municipalities). We classify as indigenous all individuals who identify themselves in the census11 as indigenous, Afro-Honduran, or black—95 percent of whom in the sample are Lenca.12 Given randomized assignment and results in appendix C that provide evidence of balance on observables across treatment arms using data from the 1988 and 2001 population censuses, our main methodological approach is to estimate a single-difference ITT model = + + + + + 1 where Yij is the outcome of interest measured in the 2013 census for individual i, born in municipality j, and G1j takes the value 1 if municipality j benefited from the (basic) CCT and 0 otherwise. β1 is the parameter of interest and yields the estimate of the long-term ITT absolute impact of past program exposure. To adhere to the experimental design, we control for the other treatment arms with indicator variables for whether municipality j benefited from both the CCT 10 While the census data contain urban or rural designation for current residential location, they do not include it for location at birth, so our original analysis plan to examine the effects of the CCT separately for urban versus rural origin locations is not possible. Instead, we distinguish between the indigenous and non-indigenous populations as described below. For other details on the proposed research design prior to data access, see Molina Millán et al. (2015), available at CCT Long-Term Impacts in Latin America: Research Proposal for Honduras. The analysis in this paper follows the research strategy outlined in that proposal, although we did not pursue analysis of Demographic and Health Survey data since it does not contain location of birth. In addition, we added analysis of spillovers to other age cohorts and added a control for the baseline outcome variable in each main specification, which in general results in more conservative estimates (see appendix A). 11 A potential concern is that the CCT might influence how people report their ethnicity. In other contexts, economic status has been shown to be associated with reported ethnicity, though this may be less salient in Honduras since the dominant indigenous group in the sample, Lenca, do not speak a different language from the rest of the population as is common for several indigenous populations in other countries in Central America. For females and males, we fail to reject the null that the probability of reporting as indigenous is unrelated to treatment status (p-values of 0.640 and 0.622, respectively). All 70 municipalities have both indigenous and nonindigenous populations. 12 Galiani, McEwan, and Quistorff (2017) provide a map of the concentration of the Lenca population in 2001, demonstrating it is the largest indigenous group both in the program area and in Honduras as a whole. 8 and supply-side interventions simultaneously (G2j) or the supply-side interventions only (G3j). Following Athey and Imbens’ (2017) recommendation to use limited and binary controls when analyzing randomized experiments, Xij includes indicator variables for the five strata used in randomization, single-year age fixed effects, and when available an indicator for whether the average value of outcome Y in municipality j for individuals born in the municipality and ages 20–25 in 2001 is above the median of the municipality-level averages for all 70 municipalities.13 Robust standard errors are calculated allowing for clustering at the municipality level. Because of the relatively small number of municipalities (40 for our principal comparison of G1 versus G4, the control), we also replicate all hypothesis tests using randomization-based inference tests (Athey and Imbens, 2017; Young, 2019). In randomization-based inference, uncertainty in the estimates arises from the random assignment of the treatments rather than from sampling. This method allows estimating the exact p-value under the sharp null hypothesis that all treatment effects are zero by calculating all possible realizations of a test statistic and rejecting if the observed realization in the experiment itself is above the significance level cutoff for the generated distribution of test statistics. Randomization-based inference provides exact finite sample test statistics without appealing to asymptotic results and as such allows testing for the influence of potential outliers and protects against accidental imbalance affecting the results. For the main analyses, we estimate ITT effects for several different age cohorts, whose selection is informed by the design and timing of the CCT. The program operated for five years (2000–2005) targeting households with pregnant women and (initially) children under three (extended to under five starting in 2003) and school-age children ages 6–12. Consequently, individuals in treatment municipalities were potentially exposed to different program components in part or in full depending on when they were born. For example, only a child born in 2000 could have directly benefitted from the nutrition and health component for the full five years. Children born a few years earlier or later, however, would have had less exposure to that 13 While the program had been in place for eight months by the time of the 2001 census, grades attained for the cohort of individuals ages 20–25 years should not have been directly affected given the program rules (and was likely to have only been minimally indirectly affected, if at all, see appendix C). At the same time, this cohort is young enough to reflect recent general secular differences in schooling in the program municipalities. Figures A.1 to A.4 replicate estimates for highest grade attained without controlling for 2001 municipality-level education and demonstrates that, if anything, the control leads to more conservative estimates. We deviate slightly from Athey and Imbens’ (2017) recommendation by not including an interaction effect of each of the binary control variables with the treatment indicators because of the limited number of municipalities assigned to each treatment arm and the large number of interactions effects this would introduce. 9 component, such as a child born in 2003 who could have directly benefited from the nutrition and health component for only two years postnatal before the program ended.14 Turning to the education component, any child 6–9 years old in 2000 could directly and fully benefit from first through fourth grade. Children older than that in 2000 also potentially benefitted, and possibly even more so if the program affected them at ages at which they otherwise might have started to drop out of school. In Figure 1, we use the short-term program evaluation data to characterize average preprogram enrollment rates for girls and boys at baseline. Patterns by age are broadly similar across the municipalities subsequently exposed to the CCT and those in the control, providing further evidence of strong balance. For both sexes, enrollment rates are above 90 percent until about age 11 after which they decline considerably. Consequently, individuals 11–13 years old in 2000 were at higher risk of dropping out when the program started; similarly, those 6–10 years old were at risk of dropping out at some point during the five years the program was in operation. Finally, individuals 14–16 years old in 2000 would not themselves have been eligible for any transfers, but nevertheless may have benefitted from transfers received by their households at ages in which their risk of dropout was high, or through peer effects. A re-analysis of the short-term program evaluation data confirms that after two years of exposure, not only cohorts directly exposed to the education conditionalities but also the older 14–16 year-old cohort (in 2000) experienced significant educational gains. Table 1 presents the ITT estimates of G1 program impacts on highest grades attained after two years. It uses the 2002 follow-up data for all children living in baseline households that by 2000 had reached primary-school age. While children with uninterrupted annual grade progression would have finished primary by ages 14–16 years, in practice more than half of the approximately 30 percent of children in this age cohort enrolled in 2000 were still enrolled in primary school (and a quarter still in the first four grades), illustrative of the accumulated schooling delays common in poor rural Honduras (Glewwe and Olinto, 2004). This also means that those children potentially would have been in the same classrooms as much younger children directly affected by the education component and conditionalities. In addition, exploration of whether effects for this ineligible cohort are concentrated in households with younger eligible siblings, via an interaction with the G1 indicator, reveal they do not seem to be. Thus, the impact on this older cohort is in line with evidence of peer effects on ineligible children in PROGRESA (Bobonis and Finan, 14 Of course a child born in 2003 also may have benefited (indirectly) in utero. 10 2009; Lalive and Cattaneo, 2009) and indicates important short-term spillovers existed for this older non-targeted age cohort. Within this context, we use the patterns of partial or full exposure to define a set of age cohorts for analysis shown in Figure 2, where for each cohort we indicate ages at the start of the program in 2000, ages in 2013 at the time of outcome measurement, and potential number of years of exposure. In addition to the age cohorts described above, with the census we can extend the window and include younger cohorts. Since outcomes are measured in 2013 and we include estimates for children born after the start of the program in 2000, going forward we report ages in 2013 as shown in the bottom row of Figure 2. In the main analyses, we estimate the impact of the basic CCT (β1) separately for each age cohort. To verify that the significance of results is not driven by multiple hypotheses testing, we compute the joint significance test of the estimated coefficients ( for all age cohorts using Young’s (2019) omnibus randomization- based inference test. Given that the experiment included three treatment arms and we estimate effects for as many as eight different age cohorts for a single outcome, for each demographic group we alternatively estimate a model combining individuals from all age cohorts relevant to the outcome being examined and directly test for the overall program impact across all age cohorts. Specifically, we extend equation (1) to include indicator variables for the age cohorts (COHORTc), taking the value 1 if individual i belongs to age cohort c, where c represents all except one of the C included cohorts. The age cohort indicator variables are also each interacted with assignment to treatment (and all other controls) yielding: = + + + + 2 + + ! × #+ For each demographic group we then implement a parametric F-test for the null hypothesis that there are no cohort-specific treatment effects in the pooled specification (Chetty, Hendren, and Katz, 2016). We compute the joint significance test of the estimated coefficients % and each of the ( % + ). That is, we test the joint significance of all of the estimated G1 treatment effects. For outcomes such as grades attained, this corresponds to the eight age cohorts shown in Figure 2. This single test on the pooled sample is not vulnerable to over-rejection rates that occur when analyzing the individual age cohorts separately. We also use Equation 2 to directly test whether impacts on grades attained differ between the cohorts with full exposure to the 11 education transfers (19–23 years old) and with full exposure to the nutrition and health components (11–12 years old). Finally, before presenting the results, we assess three selection concerns for individuals observed in the 2013 national census that might affect internal validity related to fertility, mortality, or possible undercoverage. First, as described above unintended incentives created by program eligibility rules have been linked to short-term increases in fertility in treatment areas. This is most likely for those born in the first few years of the program (i.e., ages 9–12 in 2013), after which the rules were modified. Second, if exposure to the nutrition and health components of the program reduced infant mortality, differential mortality rates are possible. Plausibly, both these forms of selectivity would increase relative cohort sizes (e.g., the ratio of children under five born to women of childbearing age) in treatment areas. We examine this possibility and find that the differences between relative cohort sizes in G1 and the control are small and insignificant for all four demographic groups (appendix Table C.5) suggesting that ITT estimates on other outcomes are unlikely to be strongly affected by fertility or mortality selection. A further potential concern is that census coverage is incomplete, with selected individuals omitted from the census for unknown reasons and possibly differentially across treatment arms. To analyze this possibility, we compare cohort sizes for those born in the 70 municipalities in the 2001 and 2013 censuses. For this exercise, we use 5-year cohorts to help neutralize differences due to age-heaping (West and Fein, 1990). Appendix Figures C.1–C.4 present the data, which include the reported international migrants. The first finding for both censuses is that within each census younger cohorts are for the most part larger than older ones, consistent with the growing Honduran population (and manifested in the empirical work below by larger sample sizes for younger cohorts), and this pattern is stronger for the indigenous populations. The figures also track cohort sizes over time, comparing the size of each cohort in 2001 with the same cohort of individuals reported 12 years later in 2013. For the 16 to 35 year olds, there are no large differences in cohort sizes between 2001 and 2013 for the four demographic groups (comparison of dark grey with blue bars).15 Differences are minimal and not systematically positive or negative for indigenous and non-indigenous women, or for indigenous men. They are a bit larger for non-indigenous men, however, possibly because of higher mortality rates (consistent with high levels of violence in rural Honduras), failure of the 2013 census to capture 15 For those below 16, 5-year old cohort sizes are not comparable between 2001 and 2013 as they were mostly born after 2001. We therefore also show the 12-15 year cohort separately. For the indigenous, the reported population size in 2001 using 2013 age is lower than in 2013, possibly due to underreporting of newborns in 2001. 12 a subset of young male adults living in Honduras16, or underestimation of international migration for this demographic group. Crucially for our analyses, however, none of the 2001–2013 differences are significantly correlated with treatment status, further alleviating potential selection concerns. 4. Results In the presentation of the results, we focus discussion on the long-term impacts of the basic CCT (G1), captured by . Appendix D presents impacts for G2 and compares them with those of G1. All ages are in 2013. All significant effects reported are robust to randomization-based inference. Specifically, randomization-based inference tests yield p-values and significance levels that are similar to the results obtained using regression-based inference tests accounting for clustering at the municipality level and all of the statistically significant point estimates reported in the figures and discussed in the text are also significantly different from zero under both methods of randomization-based inference suggested in Young (2019) (see appendix Tables A.3–A.6 for results not reported in the main tables). Appendix Table A.7 reports the pvalues from omnibus tests that combine estimates for all cohorts and outcomes examined for each demographic group, confirming the overall significance of the findings for each group.17 4.1 Education Table 2 presents the long-term impacts of the CCT on highest grade attained (defined as grades of completed schooling) by cohort and separately for females and males. For both sexes, there is clear evidence of an impact on the older cohorts fully or partially exposed to education transfers as children. Individuals 19–26 years of age in 2013 have between 0.31 and 0.43 more grades attained (compared with control group averages of about six grades or lower). While positive, effects for the younger cohorts exposed to the nutrition and health components or born after the program ended are mostly insignificant. At face value, these results suggest only minimal long-term effects on grades attained from early life exposure to the CCT. This would be a somewhat surprising conclusion, however, given the evidence on short-term effectiveness of this program for young children (Morris et al., 2004) 16 Undercounting, in part due to under-enumeration of mobile young men living in single-person households, has been documented in censuses across the world (Ewbank, 1981; Philips, Anderson, and Tsebe, 2003). 17 Table A.7 also reports omnibus joint-significance tests for all cohorts by family of outcomes (education, migration, and marriage and fertility) separately for each demographic group, confirming significance for each of the families. 13 and recent growing evidence in other contexts of the importance of investment during this early period of life. It may be that other constraints inhibit translation of the short-term effects into later improvements in education for part or all of the population. To explore this further, we consider a second potentially important dimension of heterogeneity—ethnicity. In addition to being predetermined, indigenous identity in this context may be a proxy measure for a combination of additional constraints faced by individuals during or after the program. Figures 3–12 present impacts on educational outcomes for the relevant outcomes and the four demographic groups of interest: females and males with and without indigenous backgrounds. Each figure shows the ITT effects (equation 1) of the basic CCT for individuals born in G1 municipalities, on the outcome of interest by age cohort. Point estimates are represented by blue dots with corresponding 90 (blue dash) and 95 (red square) percent regression-based confidence intervals. Figures show ages at the time of measurement in 2013, 13 years after program began, as illustrated in Figure 2. Each figure also shows the average value of the outcome in the control group (G4 municipalities) in parentheses on the x-axis label below each age cohort. Non-indigenous Females Figure 3 presents the estimates of the ITT long-term impact on grades attained for nonindigenous females. The cohort that benefited the most based on the point estimates was 19– 23 years old in 2013, i.e., those with the longest potential exposure to the G1 transfers during school-going ages. Their highest grade attained increased by more than 0.5 grades (a significant increase of nearly 10 percent). Effects are also positive and significant (about 0.4 grades) for those exposed to the nutrition and health package in early childhood, ages 11–12 and 13–15. Estimates are significant but smaller (0.2 grades) for girls 9–10 years old who were born during the program and positive but not significant for the other age cohorts. The latter include those not yet born during the program and those too old to have benefited directly from the education component. They also include girls (ages 16–18) too old at the start of the program for the nutrition and health package and too young to have directly received the full educational transfers, who in a sense fell into a gap of program coverage in the initial design. These gains in grades attained for non-indigenous females are reflected in much higher completion rates for different levels of schooling. Figure 4a presents the estimates on the probability of: (1) completing fourth grade (top-left); (2) completing primary school (i.e., sixth grade, top-right); (3) completing secondary school (i.e., 12th grade, bottom-left); and (4) having 14 started university studies (bottom-right). The impacts follow clear age patterns and show relatively large gains for the school level most relevant to each age cohort.	49,571
https://arxiv.org/abs/2403.03326	arXiv	Open Science	CC-By	null	AnatoMix: Anatomy-aware Data Augmentation for Multi-organ Segmentation	Chang Liu, Fuxin Fan, Annette Schwarz, Andreas Maier	English	Spoken	1,473	2,567	11institutetext: Pattern Recognition Lab, Friedrich-Alexander-Universität, 91058 Erlangen, Germany # AnatoMix: Anatomy-aware Data Augmentation for Multi-organ Segmentation Chang Liu 11 Fuxin Fan 11 Annette Schwarz 11 Andreas Maier 11 ###### Abstract Multi-organ segmentation in medical images is a widely researched task and can save much manual efforts of clinicians in daily routines. Automating the organ segmentation process using deep learning (DL) is a promising solution and state-of-the-art segmentation models are achieving promising accuracy. In this work, We proposed a novel data augmentation strategy for increasing the generalizibility of multi-organ segmentation datasets, namely AnatoMix. By object-level matching and manipulation, our method is able to generate new images with correct anatomy, i.e. organ segmentation mask, exponentially increasing the size of the segmentation dataset. Initial experiments have been done to investigate the segmentation performance influenced by our method on a public CT dataset. Our augmentation method can lead to mean dice of 76.1, compared with 74.8 of the baseline method. ###### Keywords: Organ Segmentation CT Data-augmentation. ## 1 Introduction Multi-organ segmentation is a widely applied clinical procedure, i.e. for treatment planning of the radiation therapy. The organs-at-risk must be delineated for planning the radiation procedure. In additional, the segmentation of the radiation sensitive organs can be applied to the estimation of CT effective dose. Deep learning (DL) based multi-organ segmentation models have achieved promising performance and the leading models in public multi-organ segmentation challenges shows that the DL-based models can have competing performance as human annotations [1]. However, DL models are usually data-demanding and a generalizable DL model needs large-scale dataset for training. On the one hand large-scale organ segmentation datasets are emerging, for example AbdomenCT-1K [3], AMOS [2] and TotalSegmentator [5]. On the other hand, many researches have been done to expand the generalizability of the segmentation dataset while keeping the robust training. In this work, we proposed a novel data augmentation strategy for multi-organ segmentation dataset inspired by human anatomy, namely AnatoMix. Regarding the task of multi-organ segmentation, the intrinsic constraints of human anatomy must be respected and the augmentation method should be able to generate anatomically correct images. Inspired by the object-level data augmentation methods and the intrinsic connection of human anatomy, we proposed AnatoMix , a organ-level data augmentation method for multi-organ segmentation tasks. AnatoMix combines the anatomical structures from different patients and thus leading to exponential increase of size of dataset. AnatoMix also accounts for the human anatomy, i.e. the correct location and size of the organs in the generated images. A major difference of AnatoMix is that it will not output unlimited augmented because the reasonable combination is also considered. Figure 1: Illustration of the concept of AnatoMix . ## 2 Method The pipeline of AnatoMix is shown in Fig. 2 , AnatoMix contains two major steps in pipeline: augmentation planning and object transplant. Experiments are designed to investigate AnatoMix on CT-ORG dataset. Figure 2: The detailed pipeline of AnatoMix . ### 2.1 Augmentation Planning The concept of AnatoMix is to separate the annotated organs from the images in the dataset, and recombine the organs onto the background image to generate new images. A segmentation dataset with $N_{d}$ images and $N_{t}$ target organs can result in $N_{d}^{N_{t}+1}$ possible combinations and each combination correspond to a new image with correct organ segmentation mask. In the case of the CT-ORG dataset, where 28 CT images with 4 annotated organs are used for training, in total 17,210,368 possible combinations can be generated. However, it is noted in our experiments that the different size and shape of patients will lead to bad combinations, for example a very large liver is matched with a patient with a small liver and the liver region will reach outside of the body region. In order to keep only the reasonable combination, a filter $f_{s}$ is created to filter out bad plans based on the normalized size of the anatomical structures $r_{s}(M_{i},M_{0})=\|\frac{\sum M_{i}-\sum M_{0}}{\sum M_{0}}\|$ (1) where $M_{0}$ and $M_{i}$ are the binary mask of the base organ and target organ. We assume that human organs will share similar shapes and thus organs with similar size will fit other patients, so the combination of organs are restrained by $r_{s}$. In our experiments, we restrain the combination by $r_{s}<0.02$ and for the case of CT-ORG dataset, the number of total combinations decreases to 1,545 after planning. ### 2.2 Object transplant For each plan, the component organs are selected for the background but the spatial location in the input image usually will not match the output image. In AnatoMix , we transplant the input organs onto the background image by spatial shift. The offset $(dx,dy,dz)$ is calculated using the binary mask $M_{i}^{}=\operatorname{arg\,max}_{d}\sum M_{b}\cdot M_{i}(dx,dy,dz),$ (2) where $M_{i}$ is the binary mask of the organ $i$ and $M_{i}(dx,dy,dz)$ shifts $M_{i}$ by $(dx,dy,dz)$. After shifting, the input mask is assured to have the biggest overlap with the background mask. And the augmented organs mask is $M^{}=\\{M_{i}^{}\|0<i<N_{t}\\}$ (3) and the output image is $I^{}=I_{b}+\sum_{0<i<N_{t}}I^{}_{i}\cdot M_{i}^{},$ (4) where $I_{b}$ is the background image and $I_{i}^{}$ is the input image of organ $i$ shifted by $(dx,dy,dz)$. ### 2.3 Data AnatoMix is evaluated on the CT-ORG dataset.[4] CT-ORG contains 120 throat- abdominal CT volumes with segmentation of lungs, liver, kidneys, bladder and bones. After filtering images with incorrect pixel-dimension, 108 images are used in the following experiments. In particular, 28 images are selected for training and 80 images for test, in order to maximally keep the robustness of the test dataset. ### 2.4 Experimental Setting Three experiments are designed to investigate the AnatoMix on different amount of training data, where 10, 20 and 28 images are used for training the segmentation model. For each experiment, we apply AnatoMix offline to augment 500 images from the base dataset, namely the atmx dataset, and then a plain U-Net is trained on each dataset, in comparison with the baseline model where no data augmentation is applied. The dice score of each organ and the average dice are used for evaluation of segmentation performance. All experiments are done on 1 Nvidia 1080 (8GB). ## 3 Results Figure 3: Some example slices of the outputs of AnatoMix . Some example slices of the outputs of AnatoMix are shown in Fig. 3. As shown in Fig. 3, AnatoMix can produce the CT volumes with correct organ location and similar organ size. From the initial results in Table. 1, AnatoMix can lead to improvement of average dice by 3.2% and 1.7% when 10 and 20 images are used for image augmentation. When 28 images are used for training, no improvement is observed using AnatoMix. Table 1: For each experiment, AnatoMix creates 500 augmented images from the corresponding 10, 20, 28 training data. The dice score of each organ is then evaluated. \| avg \| liver \| lung \| kidney \| bone ---\|---\|---\|---\|---\|--- base10 \| 0.717 \| 0.792 \| 0.948 \| 0.502 \| 0.628 atmx10 \| 0.740 \| 0.787 \| 0.944 \| 0.576 \| 0.651 base20 \| 0.748 \| 0.809 \| 0.940 \| 0.561 \| 0.681 atmx20 \| 0.761 \| 0.803 \| 0.955 \| 0.598 \| 0.689 base28 \| 0.793 \| 0.855 \| 0.961 \| 0.617 \| 0.737 atmx28 \| 0.787 \| 0.827 \| 0.961 \| 0.659 \| 0.703 ## 4 Discussion In this work, we propose AnatoMix , an object-level data augmentation method to improve the generalizability of multi-organ segmentation datasets, and investigate its performance on the CT-ORG dataset for initial evaluation. It is shown in our experiments, that AnatoMix is able to increase the segmentation performance in some cases. During our experiments, it is also noticed that AnatoMix can degrade the segmentation performance when the size of the training dataset is increased. Possible reason would be because of the intrinsic annotation errors in the CT- ORG dataset. Future research is still undergoing to test AnatoMix on other multi-organ segmentation datasets. ## References * [1] Isensee, F., Jaeger, P.F., Kohl, S.A., Petersen, J., Maier-Hein, K.H.: nnu-net: a self-configuring method for deep learning-based biomedical image segmentation. Nature methods 18(2), 203–211 (2021) * [2] Ji, Y., Bai, H., Ge, C., Yang, J., Zhu, Y., Zhang, R., Li, Z., Zhanng, L., Ma, W., Wan, X., et al.: Amos: A large-scale abdominal multi-organ benchmark for versatile medical image segmentation. Advances in NeurIPS 35, 36722–36732 (2022) * [3] Ma, J., Zhang, Y., Gu, S., Zhu, C., Ge, C., Zhang, Y., An, X., Wang, C., Wang, Q., Liu, X., et al.: Abdomenct-1k: Is abdominal organ segmentation a solved problem? IEEE PAMI 44(10), 6695–6714 (2021) * [4] Rister, B., Yi, D., Shivakumar, K., Nobashi, T., Rubin, D.L.: Ct-org, a new dataset for multiple organ segmentation in computed tomography. Scientific Data 7(1), 381 (2020) * [5] Wasserthal, J., Breit, H.C., Meyer, M.T., Pradella, M., Hinck, D., Sauter, A.W., Heye, T., Boll, D.T., Cyriac, J., Yang, S., et al.: Totalsegmentator: Robust segmentation of 104 anatomic structures in ct images. Radiology: Artificial Intelligence 5(5) (2023)	19,512
https://openalex.org/W4223983338	OpenAlex	Open Science	CC-By	2,022	YURIDIK TERMINLARNING TARJIMA QILISH XUSUSIYATLARI	Umaraliyeva Dildora Taxirjanovna	Swahili	Spoken	954	3,375	Dolzarbligi: Itimoiy hayotning shakllanishi hamisha til taraqqiyotida yetakchi qonun bo‘lgan til taraqqiyotiga xizmat qilgan. Har bir milliy tilning lug‘at tarkibi doimo harakatda, o‘zgarishda. Tilda yangi so'zlar paydo bo'ladi, ular so'z boyligini to'ldiradi va boyitadi. Hayot talabi bilan yangi so‘z tug‘iladi. Bu jarayon, ayniqsa, terminologiyada yaqqol namoyon bo‘ladi. Jamiyatda muhim ahamiyatga ega bo‘lgan termin va terminshunoslikka xalq e’tibori qaratildi. O‘zbek tilshunosligida termin va uning lingvistik xususiyatlarini o‘rganish bo‘yicha ko‘plab tadqiqotlar olib borilgan. Biroq, bu sohada hal qilinishi kerak bo'lgan ko'plab muammolar mavjud. Xususan, gidravlik terminlarning lingvistik xususiyatlari muammosi shulardan biridir. Tlshunoslikda termin tushunchasi hanuzgacha turlicha talqin qilinadi. Ba'zi tilshunoslar termin nafaqat fan va texnikaning so'z va iboralarini, balki barcha maishiy texnika nomlarini, ishlab chiqarish vositalarining nomlarini, shuningdek, o'tmishdagi ibtidoiy qurollarning barcha nomlarini, ya'ni ular ham kasbga oid so‘zlarni o‘z ichiga oladi. va amaliyot» nomli ilmiy, masofaviy, onlayn konfe YURIDIK TERMINLARNING TARJIMA QILISH XUSUSIYATLARI Umaraliyeva Dildora Taxirjanovna Oʻzbekiston xalqaro islom akademiyasi, "Chet tillar" kafedrasi oʻqituvchisi https://doi.org/10.5281/zenodo.6465829 y y, y, y YURIDIK TERMINLARNING TARJIMA QILISH XUSUSIYATLARI Umaraliyeva Dildora Taxirjanovna Oʻzbekiston xalqaro islom akademiyasi, "Chet tillar" kafedrasi oʻqituvchisi https://doi.org/10.5281/zenodo.6465829 YURIDIK TERMINLARNING TARJIMA QILISH XUSUSIYATLARI Umaraliyeva Dildora Taxirjanovna Oʻzbekiston xalqaro islom akademiyasi, "Chet tillar" kafedrasi oʻqituvchisi https://doi.org/10.5281/zenodo.6465829 ` «Zamona va amaliy ` «Zamona va amaliy «Zamonaviy dunyoda innovatsion tadqiqotlar: Nazariya va amaliyot» nomli ilmiy, masofaviy, onlayn konferensiya Tadqiqot maqsadi: “O'zbek tilining izohli lug'ati”da terminga quyidagicha izoh beriladi: “Termin (lot. Terminus – chek, chegara) fan, texnika, kasb-hunarning biror sohasiga xos muayyan bir tushunchaning aniq va barqaror ifodasi bo'lgan so'z yoki so'z birikmasi; atama”.Bizga ma'lumki, termin so'z, ibora singari tilning lug'aviy birligi hisoblanadi. Termin deb ataladigan maxsus leksik birlikni lug‘aviy birlik bo‘lgan so‘zga oddiy ko‘plik bilan qarama-qarshi qo‘yish orqali uning o‘ziga xos xususiyatlari o‘rganilmoqda. Terminologiya umumiy til leksikasi asosida shakllanadi va boyib boradi. O‘z navbatida, umumiy tilni boyitish uchun xizmat qiladi. Terminologik leksika u bilan birga jamiyat va til taraqqiyotidagi barcha jarayonlarni boshidan kechirib, xalq tili leksikasining tarkibiy qismi sifatida rivojlanadi. Tildagi maxsus terminologiya ma’lum bir soha yoki fan yuksak darajada rivojlangandagina paydo bo‘ladi. ` «Zamonaviy dunyoda innovatsion tadqiqotlar: Nazariya va amaliyot» nomli ilmiy, masofaviy, onlayn konferensiy Terminologiyaning o‘ziga xos xususiyatlari bor. Masalan, umumiy tilda sinonimiya, omonimiya, noaniqlik tilning boyligi hisoblansa, terminologiyada salbiy hodisalardir. Masalan, yarimo‘tkazgich – yarimo‘tkazgich – yarimo‘tkazgich atamalari o‘zbek tilida yagona tushunchani ifodalash uchun ishlatiladi. Bu esa o‘z navbatida o‘qitish va axborot almashish jarayonini murakkablashtiradi. Shu sababli, terminologiyasi biroz barqaror bo'lgan barcha tillarda atamalar doimiy ravishda tartibga solinadi. Tartibga solish muayyan terminologik normalarga asoslanadi. Terminologiyani rivojlantirishda fanga oid maxsus lug‘atlarning nashr etilishi ham muhim ahamiyatga ega. «Zamonaviy dunyoda innovatsion tadqiqotlar: Nazariya va amaliyot» nomli ilmiy, masofaviy, onlayn konferensiya Terminologiyaning o‘ziga xos xususiyatlari bor. Masalan, umumiy tilda sinonimiya, omonimiya, noaniqlik tilning boyligi hisoblansa, terminologiyada salbiy hodisalardir. Masalan, yarimo‘tkazgich – yarimo‘tkazgich – yarimo‘tkazgich atamalari o‘zbek tilida yagona tushunchani ifodalash uchun ishlatiladi. Bu esa o‘z navbatida o‘qitish va axborot almashish jarayonini murakkablashtiradi. Shu sababli, terminologiyasi biroz barqaror bo'lgan barcha tillarda atamalar doimiy ravishda tartibga solinadi. Tartibga solish muayyan terminologik normalarga asoslanadi. Terminologiyani rivojlantirishda fanga oid maxsus lug‘atlarning nashr etilishi ham muhim ahamiyatga ega. Material va usullar: Termin –kasb-hunarga oid bilimlar sistemasiga taalluqli tushunchaning lisoniy aks etilishidir. Uni to’g’ri tushunish uchun, termin maxsus definitsiya (izoh) talab etadi.2 Yuridik terminologiya esa (yuridik terminlar yig’indisi), har bir tilda spetifik (o’zgacha) dir. Unda yuridik til asos olgan va unda huquqning predmeti sohasidagi ma’lumotlar to’planib boradi. Yuridik terminlarga quyidagi xususiytalar xos: O’zaro uzviylik; Turg’unlik. Yuridik terminlarni tasniflash ularni sof yuridik terminlarga va umuiste’mol tildagi terminlarga bo’linishiga asoslanadi. Bu tasnif ularni o’z navbatida ikkiga bo’ladi: sof yuridik terminlar va ikki taraflama terminlar, ya’ni, ham yuridik termin, ham umumadabiy so’z. Yuridik tarjima – bu yuridik xarakterga ega bo‘lgan matnni o‘girish demakdir. Bunday matnlar sirasiga fuqarolik, sud, saylov, inson huquqlari, mol-mulk huquqiga doir matnlar kiradi. Chet el hamkorlari bilan huquqiy munosabatlar o’rnatilayotganda yuridik tarjima muhim sanaladi. Shu sababli mahorat bilan bajarilgan yuridik tarjima xalqaro yuridik munosabatlarning muvaffaqiyatli amalga oshishida muhim ajralmas bo‘lak hisoblanadi. Tomonlarning malakasi va hamkorlikning ishonchliligi tarjimaning sifati bilan belgilanadi. Natija va muhokama: Yuridik tarjima tarjimondan yetuk bilim va sinchkovlikni talab qiladi. Undagi har bir mayda detalning xatoligi katta muammolarni keltirib chiqarishi mumkin. Tarjimon birinchi navbat yuridik terminlarni mukammal bilishi talab etiladi. yuridik matnni tarjima qilayotgan tarjimon yuridika nazariyasi bilan birga boshqa davlatlardagi yuridik tizimdan ham xabardor bo‘lishi kerak. 10 10 ` «Zamonaviy dunyoda innovatsion tadqiqotlar: Nazariy va amaliyot» nomli ilmiy, masofaviy, onlayn konferens Yuridik matnlarni tarjima qilish jarayonida obyektivlik, mantiqiylik va aniqlik dominantdir. Bundan tashqari, original matndagi so‘zning tarjima tilida ekvivalenti bo‘lmasa, yoki aynan kontekstdagi ma’noni to‘liq anglata olmasa, tarjimon turli transformatsiyalardan foydalangan holda originaldagi birlikni adekvat tarzda tarjimada shunga mos birlik bilan namoyon etishi kerak. Terminologiya (termin va ... logiya) — 1) 1) leksikanutg bir sohasi; muayyan fan, texnika, ishlab chiqarish. tarmogʻining, sanʼat, ijtimoiy faoliyat sohasining tegishli tushunchalar tizimi bilan bogʻliq terminlari majmui; 2) tilshunoslikning terminlarni oʻrganuvchi sohasi. Terminologiya tor maʼnoda maʼlum bir sohaga oid maxsus leksika hisoblanadi ` «Zamonaviy dunyoda innovatsion tadqiqotlar: Nazariya va amaliyot» nomli ilmiy, masofaviy, onlayn konferensi Yuridik matnlarni tarjima qilish jarayonida obyektivlik, mantiqiylik va aniqlik dominantdir. Bundan tashqari, original matndagi so‘zning tarjima tilida ekvivalenti bo‘lmasa, yoki aynan kontekstdagi ma’noni to‘liq anglata olmasa, tarjimon turli transformatsiyalardan foydalangan holda originaldagi birlikni adekvat tarzda tarjimada shunga mos birlik bilan namoyon etishi kerak. Terminologiya (termin va ... logiya) — 1) 1) leksikanutg bir sohasi; muayyan fan, Xulosa: Xulosa qilib aytadigan bo‘lsak, terminologiyaning shakllanishi, boyitish yo‘llari har xil: boshqa tillardan so‘z olish, yangi so‘z yaratish, ayrim frazema turkumlarining leksiklashuvi, so‘z birikmalarining semantik uyg‘unlashuvi va hokazo. so‘z yasalishi va ichki so‘z yasalishi. Terminologik tizimning muayyan sohadagi barqarorligini belgilovchi muhim omil uning tartibliligi va muntazamligidir. bugungi kunda termin bilan bir qatorda, atama ham xuddi shu ma'noda qo'llaniladi. Biroq, ular terminning hozirgi ma'nosini to'liq ifoda etmaydi. Bu atama terminning xususiyatlarini aniq tasvirlab bera olmaydi. Atama terminga qaraganda torroq tushunchadir. Aniqrog‘i, bu termin bugungi kunda ilmiy tushunchalarga nisbatan qo‘llaniladi.	36,518
https://openalex.org/W4386424017	OpenAlex	Open Science	CC-By	2,023	Knockdown of ANXA10 induces ferroptosis by inhibiting autophagy-mediated TFRC degradation in colorectal cancer	Xinyuan Wang	English	Spoken	12,251	21,544	INTRODUCTION feature allows SSL to escape detection and further contributes to their progression [8]. At the molecular level, the BRAFV600E mutation and the CpG island methylator phenotype-high (CIMP-H) are the two main features of SSL and the sessile serrated carcinoma pathway [9, 10]. However, the mechanisms of SSL and serrated carcinoma pathway development remain poorly understood. Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide and the second leading cause of cancer death for both sexes [1]. In addition to the traditional adenoma- carcinoma sequence, the serrated pathway is the other major pathway leading to CRC. This pathway is characterized by the presence of a serrated lesion (SL) as a precancerous lesion [2]. However, previously, colorectal SL was misinterpreted as a harmless lesion and classiﬁed as hyperplastic polyps (HPs) [3]. In the last decade, intensive research has revealed that SL is one of the leading causes of “interstitial cancer”. Approximately 25% of sporadic CRCs develop through SL [4]. According to the latest WHO classiﬁcation of gastrointestinal tumors (5th edition) published in 2019, colorectal SL is classiﬁed into four categories: HPs, sessile serrated lesions (SSL), and sessile serrated lesions with dysplasia (SSLD), traditional serrated adenomas (TSAs) and unclassiﬁed SL [5]. Annexin A10 (ANXA10, A10) is a member of the calcium- dependent phospholipid-binding protein family, whose function has not yet been determined [11]. Several studies have found that ANXA10 is speciﬁcally highly expressed in colon SSL and MSI-H CRC [12–16], but its mechanism of action has not been investigated in depth. In other different cancers, ANXA10 has been associated with distinct roles, with high expression in some cancers, such as melanoma [17], extrahepatic cholangiocarcinoma [18], esophageal epithelial squamous cell carcinoma [19], and oral cancer [20] being associated with worse prognosis, whereas it acts as a protective factor in other cancers, such as liver cancer [21], prostate cancer [22], and bladder cancer [23]. Therefore, the role of ANXA10 in SSL and CRC needs to be further investigated. Of these, SSL is thought to have malignant potential, progressing rapidly to CRC in a short period. Two case reports have reported rapid progression to invasive tumors at 8 and 13 months, respectively [6, 7]. SSL is easily overlooked during colonoscopy as they present as pale, ill-deﬁned ﬂat lesions with a mucous surface often adhering in a “cloudy” appearance. 1Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, NHC Key Laboratory of Digestive Diseases, State Key Laboratory for Oncogenes and Related Genes, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. ✉email: lxb_1969@163.com ARTICLE OPEN Knockdown of ANXA10 induces ferroptosis by inhibiting autophagy-mediated TFRC degradation in colorectal cancer © The Author(s) 2023, corrected publication 2023 Annexin A10 (ANXA10) belongs to a family of membrane-bound calcium-dependent phospholipid-binding proteins, but its precise function remains unclear. Further research is required to understand its role in sessile serrated lesions (SSL) and colorectal cancer (CRC). We conducted transcriptome sequencing on pairs of SSL and corresponding normal control (NC) samples. Bioinformatic methods were utilized to assess ANXA10 expression in CRC. We knocked down and overexpressed ANXA10 in CRC cells to examine its effects on cell malignant ability. The effect of ANXA10 on lung metastasis of xenograft tumor cells in nude mice was also assessed. Furthermore, we used quantitative polymerase chain reaction, western blotting, and ﬂow cytometry for reactive oxygen species (ROS), lipid ROS, and intracellular Fe2+ to measure ferroptosis. Immunoblotting and Immunoﬂuorescence staining were used to detect autophagy. We found that ANXA10 was signiﬁcantly overexpressed in SSL compared to NC. ANXA10 was also highly expressed in BRAF mutant CRCs and was associated with poor prognosis. ANXA10 knockdown reduced the survival, proliferation, and migration ability of CRC cells. Knockdown of ANXA10 inhibited lung metastasis of CRC cells in mice. ANXA10 knockdown increased transferrin receptor (TFRC) protein levels and led to downregulation of GSH/GSSG, increased Fe2+, MDA concentration, and ROS and lipid ROS in cells. Knockdown of ANXA10 inhibited TFRC degradation and was accompanied by an accumulation of autophagic ﬂux and an increase in SQSTM1. Finally, Fer-1 rescued the migration and viability of ANXA10 knockdown cell lines. In brief, the knockdown of ANXA10 induces cellular ferroptosis by inhibiting autophagy-mediated TFRC degradation, thereby inhibiting CRC progression. This study reveals the mechanism of ANXA10 in ferroptosis, suggesting that it may serve as a potential therapeutic target for CRC of the serrated pathway. Cell Death and Disease (2023) 14:588 ; https://doi.org/10.1038/s41419-023-06114-2 Cell Death and Disease (2023) 14:588 ; https://doi.org/10.1038/s41419-023-06114-2 Received: 11 April 2023 Revised: 17 August 2023 Accepted: 24 August 2023 www.nature.com/cddis Antibodies and chemicals Antibodies against ANXA10 (ab213656), β-Actin (ab8227), LC3B (ab192890), and SQSTM1 (ab109012) were purchased from Abcam (USA). Rabbit anti-β-Tubulin (#2128) antibody was purchased from Cell Signaling Technology. Rabbit anti-TFRC (10084-2-AP) was purchased from Protein- tech, and mouse anti-Flag (DYKDDDDK-Tag) was purchased from Abmart. LAMP1 antibody (sc-20011) was purchased from Santa Cruz (CA, USA). Alexa Fluor 488 (A-21202) and Alexa Fluor 594 (A-21207) conjugated secondary antibodies were purchased from Invitrogen (CA, USA). Ferrostatin-1 (HY-100579), Deferoxamine mesylate (HY-B0988), Rapamycin (HY-10219), and Chloroquine (HY-17589A) were purchased from Med- ChemExpress. Cycloheximide (S7418) was purchased from Selleck, and DMSO (D2650) and N-acetyl-L-cysteine (A9165) were purchased from Sigma-Aldrich. Hydrogen peroxide solution (88597) was purchased from Millipore. Cell culture, transfection, and generating stable cell lines Cell culture, transfection, and generating stable cell lines The human colorectal cancer cell lines used in this study (HT29, HCT116, SW480, and RKO) were obtained from American Type Culture Collection (ATCC). Of these, HT29 and RKO cells carry the BRAF V600E mutation, while HCT116 and SW480 cells have the wild-type BRAF gene. HT29 and HCT116 cells were cultured in McCoy'5a medium, while SW480 and RKO cells were cultured in RPMI 1640 medium (both from Gibco, USA). In addition, 10% fetal bovine serum and 1% penicillin–streptomycin (both from Gibco, USA) were added to the respective media. All cell lines were incubated at 37 °C in a 5% CO2 atmosphere. p The day before transfection, well-grown cells were seeded onto plates following the manufacturer’s instructions. Once the cells reached 60–80% conﬂuence, small-interfering RNAs (siRNAs) and the plasmid were transfected using DharmaFECT 1 siRNA transfection reagent (Horizon Discovery Ltd., USA) and FuGene HD transfection reagent (Promega, Madison, WI, USA), respectively. The siRNAs were purchased from GenePharma Technology (Shanghai, China), while the plasmid and LV- ANXA10-shRNA were obtained from GeneChem (Shanghai, China). To achieve knockdown, Lentiviral-packed shRNA was used to infect HT29 cells with HiTransG P according to the manufacturer’s protocol. After 72 h, positive cells were selected using puromycin (8 μg/ml) for 2–3 generations to establish cell lines that were stably infected with shRNA. The plasmid and shRNA constructs used were GV492 and GV493 and the sequences of siRNAs and shRNA are listed in Table S1. To examine the coherence of samples within groups and the differences in variability between groups in RNA-seq, we employed principal component analysis (PCA) to analyze all samples. The analysis and visualization of results were carried out using the “FactoMineR” and “factorextra” R packages. For the identiﬁcation of differentially expressed genes (DEG), we conducted an enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway using the “clusterProﬁler” R package. The enrichment results were then presented graphically using the “enrichplot” R package. Patient samples This study utilized 12 pairs of sessile serrated colorectal lesions, matched normal control specimens, and colon cancer specimens obtained from Renji Hospital, Shanghai Jiao Tong University School of Medicine. The pathological diagnosis of these lesions was conducted in accordance with the latest edition of the WHO diagnostic criteria for serrated lesions in 2019 [5]. All clinical specimens underwent postoperative pathological conﬁrma- tion. Our study was approved by the Ethics Committee of Renji Hospital, Shanghai Jiao Tong University School of Medicine (KY2021-004). All patients provided informed consent. Immunohistochemistry (IHC) SSL, CRC, and normal colon tissues were obtained from Renji Hospital, Shanghai Jiao Tong University School of Medicine, and all samples were formalin-ﬁxed and embedded in parafﬁn tissue blocks. The tissue blocks were sectioned into 4 μm slices. After deparafﬁnization and antigen retrieval, the sections were incubated with 3% hydrogen peroxide solution for 20 min to quench endogenous peroxidase activity and with goat serum for 30 min to block nonspeciﬁc binding. Next, the sections were incubated overnight at 4 °C with an anti-human ANXA10 antibody (1:200) and for 30 min at room temperature with a horseradish peroxidase-conjugated secondary antibody. The sections were then visualized with 3,3-diamino- benzidine substrate and counterstained with hematoxylin before observa- tion under a light microscope. Immunoﬂuorescence staining HT29 cells were seeded at a density of 2 × 104 cells/well into a Nunc™Lab- Tek™II 8-well Chambered Slide (Thermo Fisher Scientiﬁc, Rochester, NY, USA) and transfected with siRNA after 24 h. Cells were further cultured for 48 h and then ﬁxed with 4% paraformaldehyde for 30 min. Cells were washed three times in PBS, permeabilized by 0.3% Triton X-100 for 30 min, and blocked using 1% BSA in PBS for 30 min. Cells were then incubated with primary antibodies against TFRC (1:100) and LAMP1 (1:100) overnight at 4 °C. The slide was incubated with Alexa Fluor 488- and 594-conjugated secondary antibodies for 1 h at room temperature, and followed by staining with DAPI (Southern Biotech, Birmingham, AL). Fluorescence images were photographed with FV3000 laser confocal microscope (Olympus, Japan). INTRODUCTION This g In this study, ANXA10 was identiﬁed as a molecule differentially expressed in SSL and the serrated pathway by transcriptome sequencing. The speciﬁc molecular mechanisms of ANXA10 were Ofﬁcial journal of CDDpress X. Wang et al. 2 investigated to seek out potential targets for intervention, paving the way for precise prevention and treatment of CRC. HT29 cells were dissolved in 200 μL PBS and mixed. The mice were randomly divided into two groups (n = 5 per group) and injected with the single-cell suspension through the tail vein. After eight weeks, the mice were sacriﬁced, and their lung tissues were extracted, observed, and photographed. The tissues were ﬁxed in formalin for 24 h, embedded in parafﬁn, and stained with H&E. All study procedures were approved by the Institutional Animal Care and Use Committee of Renji Hospital, School of Medicine, Shanghai Jiaotong University. Cell death, viability, and proliferation assays y p y Cell death was assessed using either the Annexin V-FITC Apoptosis Detection Kit I (BD Biosciences, USA) or the Annexin V-PE/7-AAD Apoptosis Detection Kit (Vazyme, Nanjing, China) in accordance with the manufac- turer’s instructions. The proportion of cell death was subsequently measured by ﬂow cytometry. Cell viability was determined using the Cell Counting Kit-8 (CCK-8) (Dojindo, Japan) as per the manufacturer’s recommendation. To measure cell proliferation, the CCK-8 (Dojindo, Japan) was used following the manufacturer’s instructions. Transcriptome sequencing (RNA-seq) analysis Total RNAs were extracted from SSL, paired normal tissues, HT29- shANXA10, and HT29-shNC cells using Trizol (Invitrogen, Carlsbad, CA, USA) according to the protocol. The RNAs were subsequently qualiﬁed and quantiﬁed using NanoDrop 2000 (Thermo Fisher Scientiﬁc, MA, USA). Puriﬁed mRNA was randomly fragmented into small fragments using Oligo(dT)-attached magnetic beads at the appropriate tempera- ture and subjected to PCR reactions to synthesize ﬁrst-strand cDNA and second-strand cDNA. The reaction products were then puriﬁed using magnetic beads, followed by the addition of A-Tailing Mix and RNA Index Adapters to repair the ends and subsequent PCR ampliﬁcation. The ﬁnal libraries were obtained by purifying the products using Ampure XP magnetic beads, which were then assessed for quality and quantity using an Agilent 2100 bioanalyzer before undergoing DSN treatment. The DSN-treated libraries were evaluated for quality in two ways: (i) by checking the distribution of fragment sizes using the Agilent 2100 bioanalyzer and (ii) by quantifying the libraries using qPCR. Single-end sequencing was performed on the Illumina Novaseq 6000 platform (Tsingke, Beijing, China). Single-cell transcriptome sequencing analysis g p q g y The different types of intestinal epithelial cells were clustered and grouped using the R package “Seurat” (version 4.0.3). Other analysis steps were based on our previous study [24]. Ultimately, we discovered eight signiﬁcant epithelial cell types in the SL, including a speciﬁc subpopulation g For the in vivo xenograft mouse model, 5-week-old BALB/c male nude mice were purchased from GemPharmatech Co. Ltd. (Nanjing, China) and kept in speciﬁc pathogen-free conditions at 25 °C with suitable humidity and a light/ dark cycle. To create a single-cell suspension, 1 × 106 shNC or shANXA10 Cell Death and Disease (2023) 14:588 X. Wang et al. 3 unique to the SL, which we named Epi-SL. We then compared the expression levels of ANXA10 in the eight distinct cell subpopulations. Quantitative real-time polymerase chain reaction (qPCR) Quantitative real-time polymerase chain reaction (qPCR) To extract total RNA from cells, we utilized Trizol (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The extracted RNA was assessed for quality and quantity using NanoDrop 2000 (Thermo Fisher Scientiﬁc, MA, USA), followed by reverse transcription to cDNA using a PrimeScript RT Reagent Kit (Takara, Japan). Quantitative RT-PCR was then conducted using SYBR Premix Ex Taq (Takara, Japan) in accordance with the manufacturer’s guidelines. The expression levels of mRNAs were normalized to β-Actin. Table S2 contains the sequences of all primers used in the experiment. Mass spectrometry (MS) f Mass spectrometry (MS) After protein extraction and reductive alkylation, the samples were analyzed by MS on a Thermo Fisher LTQ Qbitrap ETD Mass spectrometer. In brief, samples were placed on a Thermo Fisher Easy-nLC 1000 HPLC system equipped with a C18 column (1.8 mm, 0.15 × 1.00 mm) with solution A containing 0.1% formic acid and solution B with 100% acetonitrile. MS was performed in positive ion mode at a ﬂow rate of 300 nL/min. FTMS was used to obtain a full scan (350–1600 m/z) at a mass resolution of 30,000, followed by MS/MS using higher-energy collisional dissociation for the 10 most intense pioneer ions at 35% collisional energy and a mass resolution of 15,000. The raw MS ﬁles were analyzed using MaxQuant (1.6.1.0). The search included dynamic modiﬁcations of methionine oxidation and deamidation and static modiﬁcation of carbamidomethyl cysteine. A maximum of two missed cleavages was allowed, and the mass tolerance for precursor ions and fragment ions was set to 20 ppm and 4.5 ppm, respectively. The minimal peptide length was set to six amino acids, and a maximum of two miscleavages was allowed. The peptide and protein identiﬁcations were accepted at a false discovery rate of 0.01. For the transwell assay, 600 μL of 30% FBS medium was added to a 24- well plate, and transwell chambers (Millipore, Darmstadt, Germany) were placed on top. Next, 300 μL of cell suspension at a concentration of 1 × 106 cells/mL was added to each chamber. The cells were then incubated for 24–72 h, depending on the characteristics of the cells. After incubation, the 24-well plate was removed, and the upper chamber medium was discarded. The cells on the upper surface of the membrane were wiped off using a cotton swab, and the outer layer of the chambers was washed twice with PBS. The cells were then ﬁxed using 600 μL of 4% paraformaldehyde for 30 min, followed by staining with 600 μL of crystalline violet staining solution for 15 min at room temperature. After washing with PBS, the cells were air-dried and observed under a microscope. Six randomly selected ﬁelds of view were photographed, and the number of cells was counted. Measurement of lipid peroxidation and reactive oxygen species (ROS) Cells were pre-inoculated in 6-well plates with a density of 5 × 105 cells per well. The old medium was then removed, and 2 mL of the complete medium was added to each well. Cycloheximide (CHX) was added to each well at pre-set time points of 12th, 9th, 6th, and 0th hour according to the pre-set incubation time gradient of CHX (0, 3, 6, and 12 h). Therefore, we added 2 μL of 50 mg/mL of CHX to the corresponding wells at each pre-set time point, respectively, resulting in a ﬁnal concentration of 50 μg/mL. After a total incubation period of 12 h, all samples were subjected to protein extraction, followed by immunoblotting analysis to determine the relative gray values of the target proteins, with β-Actin expression serving as the internal reference. p Cells were inoculated in plates and transfected with siRNA after 24 h. To detect levels of lipid peroxidation, the old medium was removed after 48 h of incubation, and the cells were washed once with a serum-free medium. Then, 500 μL of a 10 μM Liperﬂuo solution (#L248, Dojindo, Japan) diluted with serum-free medium was added to each well and incubated for 30 min at 37 °C. The cells were washed twice with serum-free medium, and then digested with trypsin. After centrifuging and resuspending with 300 μL of PBS, the ﬂuorescence intensity of the samples was measured using ﬂow cytometry. y y To detect ROS levels, the old medium was removed after continuing incubation for 48 h, and 500 μL of a 10 μM DCFH-DA (#S0033, Beyotime Biotechnology, China) working solution prepared with serum-free medium was added to each well and incubated for 20 min at 37 °C. The cells were then washed three times with serum-free medium, and digested with trypsin, and the ﬂuorescence intensity of the samples was determined using ﬂow cytometry. Public database analysis y The mRNA transcriptome sequencing data for Colon Adenocarcinoma (COAD) from The Cancer Genome Atlas (TCGA, http:// cancergenome.nih.gov) and clinical information were obtained from the UCSC Xena database (https://xena.ucsc.edu/). The transcriptome data were converted into log2CPM (counts per million) for subsequent analysis using the “edgeR” R package. The colon cancer mRNA microarray dataset GSE39582, which includes 51 BRAF mutant CRC patients and 474 BRAF wild-type CRC patients, from the Gene Expression Omnibus (GEO) database was obtained using the “GEOquery” R package and annotated using the “clusterProﬁler” R package. The survival analysis of CRC patients was done using the “survival” and “survminer” R software packages, and the cutoff value for the grouping of ANXA10 expression levels was determined using the X-tile software. The relationship between gene expression levels and clinical information was processed using the “reshape2” R package and visualized using the “ggpubr” R package. Wound-healing assay and transwell assay g y y Wound-healing assays and transwell assays were performed to assess the migration capacity of the cells. For the wound-healing assay, cells were plated in 6-well plates at a density of 4 × 105 cells per well and transfected with plasmids the following day. Once the cells reached 90% conﬂuence, they were vertically scratched in the plate using a sterile 200 μL gun tip. After washing twice with PBS, the cells were cultured in a serum-free medium. Microscopic images were captured at 0 h and 40 h, and the area of the scratched region was calculated relative to the 0 h time point to determine the migration rate. Cell ferrous ion (Fe2+) level analysis y After removing the supernatant from the plate, the cells were washed three times with a serum-free medium. Subsequently, FerroOrange (1 μM, #F374, Dojindo, Japan) working solution (diluted with serum-free medium) was added and the cells were incubated for 30 min at 37 °C, as per the manufacturer’s instruction. The cells were then observed and photo- graphed directly under a ﬂuorescence microscope. Alternatively, cells were trypsinized, centrifuged at 1000 rpm for 5 min at 4 °C, and resuspended in 400 μL of serum-free medium. The ﬂuorescence intensity was immediately measured using a ﬂow cytometer. Glutathione (GSH) quantiﬁcation and malondialdehyde (MDA) assay y Intracellular levels of all glutathione and oxidized glutathione (GSSG) were detected using the GSH/GSSG Detection Kit (#S0053, Beyotime Biotech- nology, China) according to the manufacturer’s instructions. The intracel- lular MDA concentration was determined using a Cell Malondialdehyde Assay Kit (Colorimetric method, #A003-4-1, Nanjing Jiancheng Bioengi- neering Institute, China) according to the manufacturer’s recommended protocol. ANXA10 is upregulated in the serrated pathway Furthermore, ANXA10 was expressed at higher levels in CRC located in the right colon than in the left (Fig. 1F, G), consistent with the fact that SSL and BRAF-mutated CRC occur more frequently in the right. Gene set variation analysis y GSVA analysis is used to assess the enrichment of a gene set in individual samples. This analysis was performed using the “GSVA” R package, which calculates the enrichment scores, and the “ggplot2” R package was used to visualize the data. Ferroptosis occurred in ANXA10 knockdown colorectal cancer cells To elucidate the molecular mechanisms by which ANXA10 promotes malignant biological behaviors, such as survival, proliferation, and migration of CRC cells, we performed RNA-seq on HT29-shANXA10 and shNC cells. PCA reduced dimensional analysis revealed intergroup differences between the samples of the two groups, suggesting differences in transcriptome levels (Fig. 4A). Upon screening for DEG based on the criteria of differential expression fold (Fold-Change value) >2-fold or <0.5- fold and P < 0.05, we identiﬁed 153 genes with upregulated expression and 485 genes with down-regulated expression in the HT29-shANXA10 group compared to the control (Fig. 4B). KEGG pathway enrichment analysis on all up- and down-regulated genes revealed the ferroptosis pathway as the most signiﬁcant among the top 15 pathways in terms of signiﬁcance (P < 0.0001), along with other signaling pathways such as the Ras signaling pathway, arachidonic acid metabolism, NOD-like receptor signal- ing pathway, and Rap1 signaling pathway (Fig. 4C). We also performed single-cell transcriptome sequencing of serrated lesions and discovered a subpopulation of epithelial cells speciﬁc to serrated lesions (Epi-SL) (Fig. 1H) [24]. Interestingly, ANXA10 was highly expressed in Epi-SL, while its expression was lower in normal subpopulations such as absorptive epithelium and cupped cells. Immunohistochemical staining revealed that ANXA10 was speciﬁcally upregulated in the crypt at the site of lesions in SSL and BRAFmut CRC (Fig. 1I). Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells X. Wang et al. X. Wang et al. X. Wang et al. 4 We then examined the effects of ANXA10 on cell survival and apoptosis. The viability of HT29 and HCT116 was signiﬁcantly inhibited by ANXA10 siRNA (Fig. 2B, C), and a similar decrease in cell viability was observed in the HT29-shANXA10 stable- transformed cell line (Fig. 2D). Moreover, the knockdown of ANXA10 led to a signiﬁcant increase in the proportion of early and late apoptotic cells (Fig. 2E, F and S2A, B), whereas ANXA10 overexpression in SW480 and RKO cells signiﬁcantly decreased the percentage of apoptotic cells with or without cycloheximide (CHX) (Fig. 2G, H and S2C–H). ANXA10 is upregulated in the serrated pathway To investigate the features of the serrated pathway, we conducted transcriptome sequencing (RNA-seq) on twelve pairs of SSL and corresponding NC. PCA analysis revealed signiﬁcant differences between the SSL and NC groups (Fig. 1A). We performed differential analysis and identiﬁed differently expressed genes (DEG) using the \|log2FoldChange\|value > 1.5 and P < 0.05 criteria. ANXA10 was found to be signiﬁcantly upregulated in SSL (Fig. 1B). Given the high expression of ANXA10 in SSL, we further examined its expression levels in CRC patients. Analysis of the TCGA-COAD and GEO databases revealed that CRC patients with high ANXA10 expression had worse prognoses than those with low expression (P = 0.047) (Fig. 1C). In addition, ANXA10 expres- sion was substantially higher in BRAF-mutated CRC (BRAFmut CRC) than in BRAF wild-type (BRAFwt CRC) (P < 0.0001; Fig. 1D, E). Furthermore, ANXA10 was expressed at higher levels in CRC located in the right colon than in the left (Fig. 1F, G), consistent with the fact that SSL and BRAF-mutated CRC occur more frequently in the right. To investigate the features of the serrated pathway, we conducted transcriptome sequencing (RNA-seq) on twelve pairs of SSL and corresponding NC. PCA analysis revealed signiﬁcant differences between the SSL and NC groups (Fig. 1A). We performed differential analysis and identiﬁed differently expressed genes (DEG) using the \|log2FoldChange\|value > 1.5 and P < 0.05 criteria. ANXA10 was found to be signiﬁcantly upregulated in SSL (Fig. 1B). To further conﬁrm the in vitro ﬁndings, we veriﬁed the migration ability of HT29-shANXA10 cells in vitro and found that it was signiﬁcantly weaker than that of the control group (Fig. 3I, J). Subsequently, we established a lung metastasis model in nude mice by tail vein injection of HT29-shANXA10 or HT29-shNC cells. Eight weeks later, the mice were euthanized and found to have signiﬁcantly fewer metastatic lesions in the lungs of the shANXA10-injected group compared to the control group, as conﬁrmed by H&E staining (Fig. 3K–M). Given the high expression of ANXA10 in SSL, we further examined its expression levels in CRC patients. Analysis of the TCGA-COAD and GEO databases revealed that CRC patients with high ANXA10 expression had worse prognoses than those with low expression (P = 0.047) (Fig. 1C). In addition, ANXA10 expres- sion was substantially higher in BRAF-mutated CRC (BRAFmut CRC) than in BRAF wild-type (BRAFwt CRC) (P < 0.0001; Fig. 1D, E). Statistical analysis All statistical analyses were performed using GraphPad Prism 9.3.1 or R 4.1.2 software. For statistical analysis between two groups of samples, an independent sample t-test was used if they obeyed a normal distribution, while paired samples used a paired t-test. For three or more groups of samples, the one-way ANOVA was performed. Non-parametric tests were used for data that did not follow a normal distribution. All experiments were repeated three times, and differences were considered signiﬁcant if the P-value < 0.05. ANXA10 enhances the proliferation and migration of CRC cells The CCK-8 assay showed that after the knockdown of ANXA10, HT29, and HCT116 cells proliferated signiﬁcantly slower than the control group on days 3 and 4 (Fig. 3A, B). Furthermore, we observed that SW480 and RKO cells overexpressing ANXA10 proliferated substantially faster than the control cells (Fig. 3C, D). Considering the association between high ANXA10 expression and poor prognosis in CRC patients, we speculated that ANXA10 might regulate the migration ability of CRC cells. Our results demonstrated that the knockdown of ANXA10 impaired the migratory ability of CRC cells (Fig. 3E, F), while overexpression of ANXA10 promoted their migration (Fig. 3G). Scratch assays also veriﬁed that overexpression of ANXA10 facilitated the ability of SW480 cells to heal scratches (Fig. 3H). ANXA10 enhances the proliferation and migration of CRC cells The CCK-8 assay showed that after the knockdown of ANXA10, HT29, and HCT116 cells proliferated signiﬁcantly slower than the control group on days 3 and 4 (Fig. 3A, B). Furthermore, we observed that SW480 and RKO cells overexpressing ANXA10 proliferated substantially faster than the control cells (Fig. 3C, D). Flow cytometry data were analyzed using FlowJo 10.4.0 software. All images were assessed using ImageJ 1.53 software. Considering the association between high ANXA10 expression and poor prognosis in CRC patients, we speculated that ANXA10 might regulate the migration ability of CRC cells. Our results demonstrated that the knockdown of ANXA10 impaired the migratory ability of CRC cells (Fig. 3E, F), while overexpression of ANXA10 promoted their migration (Fig. 3G). Scratch assays also veriﬁed that overexpression of ANXA10 facilitated the ability of SW480 cells to heal scratches (Fig. 3H). Immunoblotting Whole cells were harvested and lysed in RIPA buffer supplemented with protease and phosphatase inhibitors. The lysates were then separated by SDS-PAGE and transferred onto nitrocellulose membranes. The mem- branes were blocked with a blocking solution, and then incubated overnight at 4 °C with primary antibodies. After washing, the membranes were incubated for 1 h at room temperature with secondary antibodies. The proteins of interest were detected by chemiluminescence using ChemiDoc Touch (Bio-Rad, California, USA). All the original western blot images were shown in supplementary information. Cell Death and Disease (2023) 14:588 Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells The names of the 15 genes with the lowest P- values are marked in the graph. C Kaplan–Meier analysis of OS revealed that the higher the level of ANXA10 expression, the worse the prognosis of CRC patients in the TCGA-COAD database. D, E ANXA10 was highly expressed in BRAFmut CRC patients compared to in BRAFwt. TCGA-COAD (D); GSE39582 (E). F, G ANXA10 was expressed at distinctly higher levels in CRC located in the right hemicolectomy than in the left. TCGA-COAD (F); GSE39582 (G). H In the serrated lesion single-cell sequencing data, ANXA10 was highly expressed in Epi-SL. I Representative images of immunohistochemical staining, with ANXA10 speciﬁcally overexpressed in the epithelial crypt at the SSL and BRAFmut CRC lesions site. The scale bars are 200 μm (10×) and 50 μm (40×). P < 0.05,P < 0.01,P < 0.001,*P < 0.0001. PCA principal component analysis, SSL sessile serrated lesion, NC normal control, OS overall survival, DEG differentially expressed genes, Epi-SL serrated lesion-speciﬁc epithelial cell. shANXA10 group (P < 0.05), and there was no signiﬁcant difference between the shANXA10 and control groups (Fig. 4J). Glutathione (GSH) is the main intracellular antioxidant, and depletion of GSH exacerbates the accumulation of intracellular ROS, ultimately triggering ferroptosis [26]. Therefore, we further examined the ratio of reduced GSH to oxidized glutathione (GSSG) in cells. The results showed that the GSH/GSSG ratio was dramatically lower in the shANXA10 cohort than in the control group, while treatment with Fer-1 signiﬁcantly restored the GSH/ GSSG ratio in HT29-shANXA10 cells (Fig. 4K). ROS levels were markedly increased after knocking down ANXA10 in CRC cells. difference between the shANXA10 and control groups (Fig. 4J). Glutathione (GSH) is the main intracellular antioxidant, and depletion of GSH exacerbates the accumulation of intracellular ROS, ultimately triggering ferroptosis [26]. Therefore, we further examined the ratio of reduced GSH to oxidized glutathione (GSSG) in cells. The results showed that the GSH/GSSG ratio was dramatically lower in the shANXA10 cohort than in the control group, while treatment with Fer-1 signiﬁcantly restored the GSH/ GSSG ratio in HT29-shANXA10 cells (Fig. 4K). The primary products of lipid peroxidation include lipid peroxide (LOOH), malondialdehyde (MDA), and 4-hydroxynonenal (4HNE), and intracellular MDA concentration can indirectly reﬂect the degree of lipid peroxidation [25]. Our experimental results showed that the MDA concentration in the HT29-shANXA10 group was soaring ahead of the control (P < 0.01). Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells To investigate the impact of ANXA10 on CRC development, we analyzed the expression levels of ANXA10 mRNA in various CRC cell lines and found the highest expression in HT29 cells, followed by HCT116 and SW480 cells, and the lowest expression in RKO cells (Fig. 2A). To study the role of ANXA10 in CRC, we performed knockdown experiments of ANXA10 in HT29 and HCT116 cells and overexpression experiments in SW480 cells. g y g g y g The considerable enrichment of the ferroptosis pathway prompted us to initiate a study on the role of ANXA10 in regulating this process. Lipid peroxidation and iron accumulation are two major biochemical features involved in ferroptosis. We, therefore, investigated the effect of knocking down ANXA10 on lipid peroxidation levels in CRC cells. Our results showed that after siRNA knockdown of ANXA10 in HT29 and HCT116 cells (siANXA10), the mean ﬂuorescence intensity was signiﬁcantly stronger detected by the liperﬂuo ﬂuorescent probe compared to the control (P < 0.01 and P < 0.05, respectively; Fig. 4D, E, H, I). In addition, we also observed an increase in intracellular reactive oxygen species (ROS) levels in HT29 cells after ANXA10 knock- down using a DCFH-DA ﬂuorescent probe (P < 0.01, Fig. 4F, G). These experimental results suggest that the lipid peroxidation and p p We used small-interfering RNAs (siRNAs) to knock down ANXA10 expression in HT29 and HCT116 cells, and siRNA-2 was identiﬁed as the most effective siRNA for ANXA10 knockdown, as conﬁrmed by RT-PCR and Western blot analysis (Fig. S1A, B). Meanwhile, we transfected ANXA10-shRNA and control lentiviral plasmids into HT29 cells to construct ANXA10 knockdown (shANAX10) and control HT29 stable-transformed cell lines (shNC), respectively, and conﬁrmed the knockdown efﬁciency by Western blot and RT-PCR analysis (Fig. S1C). Furthermore, we transiently transfected ANXA10 overexpression plasmids in SW480 and RKO cells and veriﬁed ANXA10 overexpression (Fig. S1D, E). Cell Death and Disease (2023) 14:588 X. Wang et al. ig. 1 ANXA10 is speciﬁcally overexpressed in the serrated pathway and is associated with poor prognosis. A PCA analysis of 12 pairs of SL and their corresponding NC transcriptomes sequenced showed signiﬁcant differences in transcriptome levels between them. B Volcano plot of DEG between SSL and NC. ANXA10 was signiﬁcantly overexpressed in SSL (red arrow). The names of the 15 genes with the lowest P- values are marked in the graph. Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells C Kaplan–Meier analysis of OS revealed that the higher the level of ANXA10 expression, the worse the prognosis of CRC patients in the TCGA-COAD database. D, E ANXA10 was highly expressed in BRAFmut CRC patients compared to in BRAFwt. TCGA-COAD (D); GSE39582 (E). F, G ANXA10 was expressed at distinctly higher levels in CRC located in the right hemicolectomy than in the eft. TCGA-COAD (F); GSE39582 (G). H In the serrated lesion single-cell sequencing data, ANXA10 was highly expressed in Epi-SL. Representative images of immunohistochemical staining, with ANXA10 speciﬁcally overexpressed in the epithelial crypt at the SSL and BRAFmut CRC lesions site. The scale bars are 200 μm (10×) and 50 μm (40×). P < 0.05,P < 0.01,P < 0.001,**P < 0.0001. PCA principal omponent analysis SSL sessile serrated lesion NC normal control OS overall survival DEG differentially expressed genes Epi-SL serrated 5 5 Fig. 1 ANXA10 is speciﬁcally overexpressed in the serrated pathway and is associated with poor prognosis. A PCA analysis of 12 pairs o SSL and their corresponding NC transcriptomes sequenced showed signiﬁcant differences in transcriptome levels between them. B Volcan plot of DEG between SSL and NC. ANXA10 was signiﬁcantly overexpressed in SSL (red arrow). The names of the 15 genes with the lowest P values are marked in the graph. C Kaplan–Meier analysis of OS revealed that the higher the level of ANXA10 expression, the worse th prognosis of CRC patients in the TCGA-COAD database. D, E ANXA10 was highly expressed in BRAFmut CRC patients compared to in BRAFw TCGA-COAD (D); GSE39582 (E). F, G ANXA10 was expressed at distinctly higher levels in CRC located in the right hemicolectomy than in th left. TCGA-COAD (F); GSE39582 (G). H In the serrated lesion single-cell sequencing data, ANXA10 was highly expressed in Epi-S I Representative images of immunohistochemical staining, with ANXA10 speciﬁcally overexpressed in the epithelial crypt at the SSL an BRAFmut CRC lesions site. The scale bars are 200 μm (10×) and 50 μm (40×). P < 0.05,P < 0.01,P < 0.001,**P < 0.0001. PCA princip Fig. 1 ANXA10 is speciﬁcally overexpressed in the serrated pathway and is associated with poor prognosis. A PCA analysis of 12 pairs of SSL and their corresponding NC transcriptomes sequenced showed signiﬁcant differences in transcriptome levels between them. B Volcano plot of DEG between SSL and NC. ANXA10 was signiﬁcantly overexpressed in SSL (red arrow). Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells Notably, treating cells with the ferroptosis inhibitor Ferrostatin-1 (Fer-1) for 48 h signiﬁcantly reduced the MDA concentration in the Cell Death and Disease (2023) 14:588 Fig. 2 Knockdown ANXA10 reduced colorectal cancer cell survival and promoted apoptosis. A Expression levels of ANXA10 mRNA in different CRC cell lines. B–D CCK-8 assay to detect the survival rate of HT29 (B) and HCT116 (C) cells after transfection with siRNA and HT29 cells (D) after infection with lentiviral-packed shRNA. E, F Flow plots of the change in the percentage of apoptosis (E) and statistical results (F) of the percentage of early apoptosis + late apoptosis (Q2 + Q3) after knockdown of HT29 cells using ﬂow cytometry. G, H Flow plots of the change in the percentage of apoptosis (G) and statistical results (H) of the percentage of early apoptosis + late apoptosis (Q2 + Q3) after overexpression of ANXA10 and treatment with 5 μg/ml CHX for 24 h in SW480 cells using ﬂow cytometry. P < 0.05, *P < 0.01. CCK-8 Cell Counting Kit-8; CHX cycloheximide. X. Wang et al. X. Wang et al. 6 Fig. 2 Knockdown ANXA10 reduced colorectal cancer cell survival and promoted apoptosis. A Expression levels of ANXA10 mRNA in different CRC cell lines. B–D CCK-8 assay to detect the survival rate of HT29 (B) and HCT116 (C) cells after transfection with siRNA and HT29 cells (D) after infection with lentiviral-packed shRNA. E, F Flow plots of the change in the percentage of apoptosis (E) and statistical results (F) of the percentage of early apoptosis + late apoptosis (Q2 + Q3) after knockdown of HT29 cells using ﬂow cytometry. G, H Flow plots of the change in the percentage of apoptosis (G) and statistical results (H) of the percentage of early apoptosis + late apoptosis (Q2 + Q3) after overexpression of ANXA10 and treatment with 5 μg/ml CHX for 24 h in SW480 cells using ﬂow cytometry. P < 0.05, *P < 0.01. CCK-8 Cell Counting Kit-8; CHX cycloheximide. Fig. 4N, O). Finally, we observed the ﬂuorescence images of the FerroOrange probe inside the cells and noticed a distinct decrease in ﬂuorescence intensity in the shANXA10 group of cells after DFO treatment (Fig. 4P). Subsequently, we detected the levels of Fe2+ in CRC cells using the FerroOrrange ﬂuorescent probe. Knockdown of ANXA10 represses viability and promotes the death of colorectal cancer cells We found that the mean ﬂuorescence intensity was more robust in the siANXA10 group than in the control (P < 0.001). After treating both groups of cells separately with the iron chelator, deferoxamine mesylate (DFO), for 48 h, we found that DFO dramatically reduced the mean ﬂuorescence intensity of cells in the siANXA10 group (P < 0.0001, Fig. 4L, M). Therefore, we again examined Fe2+ levels in HT29 cells by ﬂow cytometry and found experimental results consistent with those described above. Knockdown of ANXA10 resulted in a pronounced increase in intracellular mean ﬂuorescence intensity (P < 0.01), while DFO signiﬁcantly restored this trend (P < 0.01, Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation g To dissect the speciﬁc mechanisms underlying ferroptosis, we analyzed changes in ferroptosis-related genes in HT29 cells with ANXA10 knockdown. Ligase Catalytic Subunit (GCLC), Glutamate- Cysteine Ligase Modiﬁer Subunit (GCLM), Solute Carrier Family 39 Member 14 (SLC39A14/Zip14), and Ceruloplasmin (CP) were Cell Death and Disease (2023) 14:588 X. Wang et al. Fig. 3 ANXA10 promoted the proliferation and migration of colorectal cancer cells. A, B CCK-8 assay to determine the level of proliferation and found decreased proliferation capacity after knockdown of ANXA10 in HT29 (A) and HCT116 (B) cells. C, D CCK-8 assay to determine the level of proliferation and found increased proliferation capacity after overexpression of ANXA10 in SW480 (C) and RKO (D) cells. E, F Transwell migration assay to detect the effect of knockdown of ANXA10 on the migration ability of HCT116 (E) and HT29 (F) cells. G Transwell migration assay to detect the effect of overexpression of ANXA10 on the migratory ability of SW480 cells. Six randomly selected ﬁelds of view were photographed for each group; representative images are shown on the left, the scale bar is 100 μm; and statistical results are shown on the right (n = 6). H Scratch assay was performed to detect the migration ability of SW480 in control and overexpression groups. Representative images are shown on the left, scale bar is 200 μm; the migration rates of cells in each group are shown on the right. I, J Transwell migration assay was performed to detect the migration ability of the HT29-shANXA10 cell line and control group. Representative images at a scale of 200 μm (I) and the statistical results (n = 6) (J) were shown. K–M A lung metastasis model was constructed by injecting HT29-shANXA10 and controls into nude mice. Representative lung tissue images (K) and H&E-stained images of lung metastases (L) were illustrated at a scale of 200 μm for 10× images and 50 μm for 40× images; the statistical analysis of the number of lung metastases in the two groups is shown (M) (n = 5). P < 0.05, P < 0.01,P < 0.001, **P < 0.0001. 7 7 Fig. 3 ANXA10 promoted the proliferation and mig Fig. 3 ANXA10 promoted the proliferation and migration of colorectal cancer cells. A, B CCK-8 assay to determine the level of proliferation and found decreased proliferation capacity after knockdown of ANXA10 in HT29 (A) and HCT116 (B) cells. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation C, D CCK-8 assay to determine the level of proliferation and found increased proliferation capacity after overexpression of ANXA10 in SW480 (C) and RKO (D) cells. E, F Transwell migration assay to detect the effect of knockdown of ANXA10 on the migration ability of HCT116 (E) and HT29 (F) cells. G Transwell migration assay to detect the effect of overexpression of ANXA10 on the migratory ability of SW480 cells. Six randomly selected ﬁelds of view were photographed for each group; representative images are shown on the left, the scale bar is 100 μm; and statistical results are shown on the right (n = 6). H Scratch assay was performed to detect the migration ability of SW480 in control and overexpression groups. Representative images are shown on the left, scale bar is 200 μm; the migration rates of cells in each group are shown on the right. I, J Transwell migration assay was performed to detect the migration ability of the HT29-shANXA10 cell line and control group. Representative images at a scale of 200 μm (I) and the statistical results (n = 6) (J) were shown. K–M A lung metastasis model was constructed by injecting HT29-shANXA10 and controls into nude mice. Representative lung tissue images (K) and H&E-stained images of lung metastases (L) were illustrated at a scale of 200 μm for 10× images and 50 μm for 40× images; the statistical analysis of the number of lung metastases in the two groups is shown (M) (n = 5). P < 0.05, P < 0.01,P < 0.001, **P < 0.0001. Fig. 3 ANXA10 promoted the proliferation and migration of colorectal cancer cells. A, B CCK-8 assay Fig. 3 ANXA10 promoted the proliferation and migration of colorectal cancer cells. A, B CCK-8 assay to determine the level of proliferation and found decreased proliferation capacity after knockdown of ANXA10 in HT29 (A) and HCT116 (B) cells. C, D CCK-8 assay to determine the level of proliferation and found increased proliferation capacity after overexpression of ANXA10 in SW480 (C) and RKO (D) cells. E, F Transwell migration assay to detect the effect of knockdown of ANXA10 on the migration ability of HCT116 (E) and HT29 (F) cells. G Transwell migration assay to detect the effect of overexpression of ANXA10 on the migratory ability of SW480 cells. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation Six randomly selected ﬁelds of view were photographed for each group; representative images are shown on the left, the scale bar is 100 μm; and statistical results are shown on the right (n = 6). H Scratch assay was performed to detect the migration ability of SW480 in control and overexpression groups. Representative images are shown on the left, scale bar is 200 μm; the migration rates of cells in each group are shown on the right. I, J Transwell migration assay was performed to detect the migration ability of the HT29-shANXA10 cell line and control group. Representative images at a scale of 200 μm (I) and the statistical results (n = 6) (J) were shown. K–M A lung metastasis model was constructed by injecting HT29-shANXA10 and controls into nude mice. Representative lung tissue images (K) and H&E-stained images of lung metastases (L) were illustrated at a scale of 200 μm for 10× images and 50 μm for 40× images; the statistical analysis of the number of lung metastases in the two groups is shown (M) (n = 5). P < 0.05, P < 0.01,P < 0.001, **P < 0.0001. cell membrane [28] while SLC39A14 is involved in the uptake of cellular non-transferrin-bound iron [29, 30]. elevated in HT29-shANXA10 cells, while SLC40A1 and SLC11A2/ DMT1 were reduced (Fig. 5A). TFRC is a crucial bridge involved in translocating transferrin (TF)-bound iron (III) into cells [27]. Fe3+ entering the cell is reduced to Fe2+ by STEAP3, a metal reductase in the endosome, and then transported to the cytoplasm by SLC11A2. SLC40A1 is the primary channel for Fe2+ export from the To conﬁrm the changes in iron ion transport-related genes, we examined their mRNA levels in HT29-shANXA10 and ANXA10- knockdown HCT116 cells (Fig. 5B and S3). qPCR results showed that the expression level of TFRC (P < 0.05 and P < 0.001 in HT29 Cell Death and Disease (2023) 14:588 X. Wang et al. 8 ig. 4 Knockdown of ANXA10 induced ferroptosis in colorectal cancer cells. A PCA analysis of HT29-shANXA10 and shNC transcriptome equencing. B Volcano map of differentially expressed genes. The names of the 20 genes with the largest \|log2FoldChange\| values are marked. C Results of KEGG pathway enrichment analysis. D, E After the transfection of siRNA, lipid peroxidation levels were detected in HT29 using perﬂuo ﬂuorescent probes. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation H, I After the transfection of siRNA, lipid peroxidation levels were detected in HCT116 cells using liperﬂuo uorescent probes. F, G Detection of ROS levels in HT29 cells using DCFH-DA ﬂuorescent probe. Cells were treated with 400 μM H2O2 for 2 h as positive control and with 400 μM H2O2 and 8 mM NAC as a negative control. J–K Intracellular MDA levels (J) and the ratio of reduced GSH/ GSSG (K) were measured in HT29-shANXA10 and control cells after treating the cells with Fer-1 (1 μM) or DMSO for 48 h, respectively. –O Intracellular Fe2+ levels in HCT116 (L, M) and HT29 cells (N, O) were measured using FerroOrange ﬂuorescent probes after transfection of iRNA knockdown ANXA10 or retreatment of cells with DFO (200 μM) for 48 h, respectively. P Fluorescence images of the intracellular erroOrange probe in the HT29-shANXA10 and shNC groups at a scale of 100 μM. P < 0.05,P < 0.01,P < 0.001,**P < 0.0001. PCA principal omponent analysis, ROS oxygen species, MDA malondialdehyde, GSSG oxidized glutathione, GSH glutathione, DFO deferoxamine mesylate, er-1 ferrostatin-1, NAC N-acetyl-L-cysteine, H2O2 hydrogen peroxide. Fig. 4 Knockdown of ANXA10 induced ferroptosis in colorectal cancer cells. A PCA analysis of HT29-shANXA10 and shNC transcriptome sequencing. B Volcano map of differentially expressed genes. The names of the 20 genes with the largest \|log2FoldChange\| values are marked. C Results of KEGG pathway enrichment analysis. D, E After the transfection of siRNA, lipid peroxidation levels were detected in HT29 using liperﬂuo ﬂuorescent probes. H, I After the transfection of siRNA, lipid peroxidation levels were detected in HCT116 cells using liperﬂuo ﬂuorescent probes. F, G Detection of ROS levels in HT29 cells using DCFH-DA ﬂuorescent probe. Cells were treated with 400 μM H2O2 for 2 h as a positive control and with 400 μM H2O2 and 8 mM NAC as a negative control. J–K Intracellular MDA levels (J) and the ratio of reduced GSH/ GSSG (K) were measured in HT29-shANXA10 and control cells after treating the cells with Fer-1 (1 μM) or DMSO for 48 h, respectively. L–O Intracellular Fe2+ levels in HCT116 (L, M) and HT29 cells (N, O) were measured using FerroOrange ﬂuorescent probes after transfection of siRNA knockdown ANXA10 or retreatment of cells with DFO (200 μM) for 48 h, respectively. P Fluorescence images of the intracellular FerroOrange probe in the HT29-shANXA10 and shNC groups at a scale of 100 μM. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation P < 0.05,P < 0.01,P < 0.001,**P < 0.0001. PCA principal component analysis, ROS oxygen species, MDA malondialdehyde, GSSG oxidized glutathione, GSH glutathione, DFO deferoxamine mesylate, Fer-1 ferrostatin-1, NAC N-acetyl-L-cysteine, H2O2 hydrogen peroxide. knockdown of ANXA10 inhibits the degradation of TFRC protein in HT29 cells. and HCT116 cells, respectively), which mediates the inward ﬂow of iron ions, was signiﬁcantly increased in the ANXA10-knockdown groups of cells, whereas the expression levels of SLC40A1 (P < 0.05 in both HT29 and HCT116 cells), which translated the inward ﬂow of iron ions, had slightly lower expression levels. Next, we investigated the expression levels of TFRC, an essential protein that is the dominant mode of iron uptake for cells. Western blot assays conﬁrmed higher TFRC protein expression in HT29- shANXA10 cells than in shNC cells. However, the increase in TFRC expression was not restored after 48 h treatment with Fer-1 (1 μM) or DFO (200 μM) (Fig. 5C). The half-life of TFRC protein was signiﬁcantly longer in HT29-shANXA10 cells than in shNC cells after inhibition of intracellular protein synthesis by cycloheximide (CHX) treatment (P < 0.05, Fig. 5D, E). Therefore, we speculate that increased levels of TFRC expression may be responsible for the ferroptosis that occurs in HT29-shANXA10 cells, and the and HCT116 cells, respectively), which mediates the inward ﬂow of iron ions, was signiﬁcantly increased in the ANXA10-knockdown groups of cells, whereas the expression levels of SLC40A1 (P < 0.05 in both HT29 and HCT116 cells), which translated the inward ﬂow of iron ions, had slightly lower expression levels. Next, we investigated the expression levels of TFRC, an essential protein that is the dominant mode of iron uptake for cells. Western blot assays conﬁrmed higher TFRC protein expression in HT29- shANXA10 cells than in shNC cells. However, the increase in TFRC expression was not restored after 48 h treatment with Fer-1 (1 μM) or DFO (200 μM) (Fig. 5C). The half-life of TFRC protein was signiﬁcantly longer in HT29-shANXA10 cells than in shNC cells after inhibition of intracellular protein synthesis by cycloheximide (CHX) treatment (P < 0.05, Fig. 5D, E). Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation Therefore, we speculate that increased levels of TFRC expression may be responsible for the ferroptosis that occurs in HT29-shANXA10 cells, and the Research has revealed that TFRC degradation can occur through autophagy [31, 32], and based on this, we hypothesized that inhibiting autophagy-mediated degradation through the knock- down of ANXA10 may prevent the degradation of TFRC. To test our conjecture, we examined LC3B and autophagy receptor SQSTM1 protein levels. We found that HT29-shANXA10 cells accumulated signiﬁcantly higher levels of LC3B compared to control cells, with no change in trend after treatment with either the autophagy inducer rapamycin (Rap) or the autophagy inhibitor chloroquine (CQ) (Fig. 5F). The protein expression level of SQSTM1 was signiﬁcantly higher in HT29-shANXA10 cells than in the control group (Fig. 5F, G). The immunoﬂuorescence analysis showed that TFRC was co-localized with the lysosomal marker LAMP1 in HT29 cells, but the co-localization of TFRC and LAMP1 Cell Death and Disease (2023) 14:588 X. Wang et al. Fig. 5 Knockdown of ANXA10-induced ferroptosis through inhibiting autophagy-mediated TFRC degradation. A Heatmap showing the expression of genes related to ferroptosis in RNA-seq. The redder the color of the box, the higher the expression of a gene in that sample; conversely, the bluer the color, the lower the expression. B qPCR to detect the expression levels of SLC40A1, SLC39A14, SLC11A2, TFRC, and FTH1 mRNA in HT29-shANXA10 and control groups of cells (n = 3). C Protein expression levels of TFRC in HT29-shANXA10 cells and control were compared by Western blot assay after 48 h incubation in complete medium, complete medium containing DFO (200 μM) or Fer-1 (1 μM). D, E Western blot assay comparing the protein expression levels of intracellular TFRC in shANXA10 HT29 cells and control after treatment with CHX (50 μg/mL) for 0, 3, 6, and 12 h, respectively (D). The TFRC/β-Actin ratio was calculated for both groups after 3, 6, and 12 h of CHX treatment, respectively, using the 0 h group as a reference (E). F Western blot assay comparing the protein expression levels of LC3B in HT29- shANXA10 cells and control group after 48 h incubation in DMSO (1:1000), Rap (200 nM), or CQ (50 μM). G Western blot assay comparing the protein expression levels of SQSTM1 and TFRC in HT29-shANXA10 cells and controls. H HT29 cells transfected with control or ANXA10 siRNAs were used for co-localization analysis for LAMP1 and TFRC. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation Yellow ﬂuorescence in merged images indicates their co-localization. Scale bars = 20 μm. I Results of PCA analysis of the proteomics of HT29-shANXA10 and control groups. J Volcano plot of DEP. The names of the 20 proteins with the largest \|log2FoldChange\|values are marked. K Correlation analysis of transcriptomic and proteomic results. The horizontal and the vertical coordinates are the results of taking the logarithm of the fold-change in mRNA and protein expression levels, respectively. L Correlation analysis of results for statistically signiﬁcant features identiﬁed by GSVA analysis of transcriptomics and proteomics separately for hallmark. The horizontal and the vertical coordinates are the results of logarithmic ploidy change in transcriptomic and proteomic sequencing, respectively. P < 0.05, P < 0.01, P < 0.001, ns no signiﬁcance. DEP differentially expressed proteins, DFO deferoxamine mesylate, Fer-1 Ferrostatin-1, Rap rapamycin, CQ chloroquine, CHX cycloheximide. g 9 Fig. 5 Knockdown of ANXA10-induced ferroptosis through inhibiting autophagy-mediated TFRC degradation. A Heatmap showing the expression of genes related to ferroptosis in RNA-seq. The redder the color of the box, the higher the expression of a gene in that sample; conversely, the bluer the color, the lower the expression. B qPCR to detect the expression levels of SLC40A1, SLC39A14, SLC11A2, TFRC, and FTH1 mRNA in HT29-shANXA10 and control groups of cells (n = 3). C Protein expression levels of TFRC in HT29-shANXA10 cells and control were compared by Western blot assay after 48 h incubation in complete medium, complete medium containing DFO (200 μM) or Fer-1 (1 μM). D, E Western blot assay comparing the protein expression levels of intracellular TFRC in shANXA10 HT29 cells and control after treatment with CHX (50 μg/mL) for 0, 3, 6, and 12 h, respectively (D). The TFRC/β-Actin ratio was calculated for both groups after 3, 6, and 12 h of CHX treatment, respectively, using the 0 h group as a reference (E). F Western blot assay comparing the protein expression levels of LC3B in HT29- shANXA10 cells and control group after 48 h incubation in DMSO (1:1000), Rap (200 nM), or CQ (50 μM). G Western blot assay comparing the protein expression levels of SQSTM1 and TFRC in HT29-shANXA10 cells and controls. H HT29 cells transfected with control or ANXA10 siRNAs were used for co-localization analysis for LAMP1 and TFRC. Yellow ﬂuorescence in merged images indicates their co-localization. Scale bars = 20 μm. Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation I Results of PCA analysis of the proteomics of HT29-shANXA10 and control groups. J Volcano plot of DEP. The names of the 20 proteins with the largest \|log2FoldChange\|values are marked. K Correlation analysis of transcriptomic and proteomic results. The horizontal and the vertical coordinates are the results of taking the logarithm of the fold-change in mRNA and protein expression levels, respectively. L Correlation analysis of results for statistically signiﬁcant features identiﬁed by GSVA analysis of transcriptomics and proteomics separately for hallmark. The horizontal and the vertical coordinates are the results of logarithmic ploidy change in transcriptomic and proteomic sequencing, respectively. P < 0.05, P < 0.01, P < 0.001, ns no signiﬁcance. DEP differentially expressed proteins, DFO deferoxamine mesylate, Fer-1 Ferrostatin-1, Rap rapamycin, CQ chloroquine, CHX cycloheximide. (Pearson r = 0.38, P < 0.001, Fig. 5J). Interestingly, SQSTM1 and TFRC were among the 33 genes with signiﬁcant changes in both histological sequencing results (P < 0.05) (Fig. 5K), conﬁrming that knockdown of ANXA10 led to the accumulation of SQSTM1 and suppression of TFRC degradation in CRC cells. Moreover, correla- tion analysis of transcriptomic and proteomic signiﬁcantly enriched Hallmark pathways revealed that characteristic pathways such as tumor necrosis factor α (TNF-α)/NFκB signaling, apoptosis, Hedgehog signaling, protein secretion, and cholesterol home- ostasis were signiﬁcantly enriched in the shANXA10 group, while pathways such as interferon γ responsiveness, interferon α responsiveness, and bile acid metabolism were downmodulated (Fig. 5L). s C f - , - y s s s 0 s t (Pearson r = 0.38, P < 0.001, Fig. 5J). Interestingly, SQSTM1 and TFRC were among the 33 genes with signiﬁcant changes in both histological sequencing results (P < 0.05) (Fig. 5K), conﬁrming that knockdown of ANXA10 led to the accumulation of SQSTM1 and suppression of TFRC degradation in CRC cells. Moreover, correla- tion analysis of transcriptomic and proteomic signiﬁcantly enriched Hallmark pathways revealed that characteristic pathways such as tumor necrosis factor α (TNF-α)/NFκB signaling, apoptosis, Hedgehog signaling, protein secretion, and cholesterol home- ostasis were signiﬁcantly enriched in the shANXA10 group, while pathways such as interferon γ responsiveness, interferon α responsiveness, and bile acid metabolism were downmodulated (Fig. 5L). Cell viability and migration were rescued by Ferrostatin-1 in ANXA10 knockdown colorectal cancer cells Owing to the above observations, we hypothesized that the knockdown of ANXA10 induces ferroptosis in CRC cells by decreased after ANXA10 knockdown (Fig. 5H). Knockdown of ANXA10 inhibits autophagy-mediated TFRC degradation These results suggest that induction of autophagy partially restored TFRC degradation in HT29-shANXA10 cells, implying that knockdown of ANXA10 prevented TFRC degradation by inhibiting the autopha- gic process, especially the degradation of TFRC in the lysosome, thereby inducing ferroptosis in CRC cells (Fig. 6G). y g p g To gain a more comprehensive understanding of the ANXA10- regulated protein proﬁle, we performed mass spectrometry analysis on HT29-shANXA10 cells and controls. PCA analysis showed a distinguished difference in protein expression proﬁles between the two groups (Fig. 5I). Differentially expressed proteins (DEP) were screened according to the criteria of differential expression fold (Fold-Change value) >2-fold or <0.5-fold and P < 0.05. 46 proteins with upregulated expression and 40 with down-regulated expression were identiﬁed in the HT29-shANXA10 group compared to the control group (Fig. 5J). Correlation analysis of transcriptome and proteome sequencing results revealed that their changes were positively correlated after ANXA10 knockdown Cell Death and Disease (2023) 14:588 Cell viability and migration were rescued by Ferrostatin-1 in ANXA10 knockdown colorectal cancer cells Owing to the above observations, we hypothesized that the knockdown of ANXA10 induces ferroptosis in CRC cells by Cell Death and Disease (2023) 14:588 nhibiting autophagy-mediated TFRC degradation thereby sup- control cells Fer-1 greatly improved the migration ability of g. 6 Ferroptosis inhibitor restored viability and migration of colorectal cancer cells with ANXA10 knockdown. A CCK-8 assay to detect he cell viability of HCT116 siANXA10 and siNC cells after 48 h incubation in a complete medium containing DMSO (1:1000) and Fer-1 (1 μM), espectively. B, C Transwell migration assay to detect the effect of Fer-1 (1 μM) on the migration ability of HCT116 siANXA10 cells and siNC ells. D CCK-8 assay to determine the cell viability of HT29 siA10 and siNC cells after 48 h incubation in a complete medium containing DMSO :1000) and Fer-1 (1 μM) respectively. E, F Transwell migration assay to detect the effect of Fer-1 (1 μM) on the migration ability of T29 siANXA10 cells and siNC cells. Five ﬁelds of view were randomly photographed in each group at a scale of 200 μm (n = 5). G The chematic representation of ferroptosis induced by knockdown of ANXA10 through inhibition of autophagy-mediated TFRC degradation. P < 0.05, P < 0.01, P < 0.001, *P < 0.0001, ns no signiﬁcance. X. Wang et al. X. Wang et al. 10 Fig. 6 Ferroptosis inhibitor restored viability and migration of colorectal cancer cells with ANXA10 knockdown. A CCK-8 assay to detect the cell viability of HCT116 siANXA10 and siNC cells after 48 h incubation in a complete medium containing DMSO (1:1000) and Fer-1 (1 μM), respectively. B, C Transwell migration assay to detect the effect of Fer-1 (1 μM) on the migration ability of HCT116 siANXA10 cells and siNC cells. D CCK-8 assay to determine the cell viability of HT29 siA10 and siNC cells after 48 h incubation in a complete medium containing DMSO (1:1000) and Fer-1 (1 μM) respectively. E, F Transwell migration assay to detect the effect of Fer-1 (1 μM) on the migration ability of HT29 siANXA10 cells and siNC cells. Five ﬁelds of view were randomly photographed in each group at a scale of 200 μm (n = 5). G The schematic representation of ferroptosis induced by knockdown of ANXA10 through inhibition of autophagy-mediated TFRC degradation. P < 0.05, P < 0.01, P < 0.001, ***P < 0.0001, ns no signiﬁcance. REFERENCES 1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality World- wide for 36 Cancers in 185 Countries. Cancer J Clin. 2021;71:209–49. 2. Leggett B, Whitehall V. Role of the serrated pathway in colorectal cancer pathogenesis. Gastroenterology. 2010;138:2088–100. 3. Longacre TA, Fenoglio-Preiser CM. Mixed hyperplastic adenomatous polyps/ser- rated adenomas. A distinct form of colorectal neoplasia. Am J Surg Pathol. 1990;14:524–37. Research on ferroptosis is expanding and showing potential for the development of new therapeutic approaches for CRC. For example, salazosulfapyridine can induce ferroptosis by inhibiting the cell membrane cystine glutamate transporter receptor (System Xc-) and depleting GSH, which promotes sensitivity to cisplatin in CRC [34]. Dimethyl fumarate induces ferroptosis in CRC cells by depleting GSH, increasing ROS, and activating MAPK [35]. Targeting ferroptosis can enhance the therapeutic effect of CRC, improve sensitivity to radiotherapy, and reduce cellular resistance [36]. Our study found that knockdown of ANXA10 induced ferroptosis in CRC cells, suggesting that ANXA10 is an effective target for inducting ferroptosis in the treatment of ANXA10 highly expressed CRC. 4. Crockett SD, Nagtegaal ID. Terminology, molecular features, epidemiology, and management of serrated colorectal neoplasia. Gastroenterology. 2019;157:949–66.e4. 5. Nagtegaal ID, Odze RD, Klimstra D, Paradis V, Rugge M, Schirmacher P, et al. The 2019 WHO classiﬁcation of tumours of the digestive system. Histopathology. 2020;76:182–8. 6. Oono Y, Fu K, Nakamura H, Iriguchi Y, Yamamura A, Tomino Y, et al. Progression of a sessile serrated adenoma to an early invasive cancer within 8 months. Dig Dis Sci. 2009;54:906–9. 7. Amemori S, Yamano H-O, Tanaka Y, Yoshikawa K, Matsushita H-O, Takagi R, et al. Sessile serrated adenoma/polyp showed rapid malignant transformation in the ﬁnal 13 months. Dig Endosc. 2020;32:979–83. 8. Murakami T, Sakamoto N, Nagahara A. Endoscopic diagnosis of sessile serrated adenoma/polyp with and without dysplasia/carcinoma. World J Gastroenterol. 2018;24:3250–9. Iron deﬁciency in CRC has been linked to a less favorable prognosis and poor treatment response [37, 38]. Therefore, increasing the concentration of Fe2+ in CRC cells is expected to improve the prognosis of CRC patients. Our study revealed that knockdown of ANXA10 inhibited the degradation of TFRC, resulting in a signiﬁcant increase in intracellular Fe2+ concen- tration and inducing ferroptosis in CRC cells. Thus, it is possible to signiﬁcantly inhibit cell growth by up-regulating the concentration of iron ions in ANXA10 highly expressed CRC cells. 9. REFERENCES De Palma FDE, D’Argenio V, Pol J, Kroemer G, Maiuri MC, Salvatore F. The molecular hallmarks of the serrated pathway in colorectal cancer. Cancers (Basel). 2019;11:1017. 10. Tao Y, Kang B, Petkovich DA, Bhandari YR, In J, Stein-O'Brien G. et al. Aging-like spontaneous epigenetic silencing facilitates Wnt activation, stemness, and BrafV600E-induced tumorigenesis. Cancer Cell. 2019;35:315–28.e6. 11. Xi Y, Ju R, Wang Y. Roles of Annexin A protein family in autophagy regulation and therapy. Biomed Pharmacother. 2020;130:110591. 12. Macaron C, Lopez R, Pai RK, Burke CA. Expression of annexin A10 in serrated polyps predicts the development of metachronous serrated polyps. Clin Transl Gastroenterol. 2016;7:e205. In our study, we observed a distinct increase in autophagic ﬂux and accumulation of SQSTM1 following knockdown of ANXA10, which ultimately hindered the autophagic process in CRC cells, suggesting that ANXA10 plays a role in the autophagy process. This biological function is similar to other proteins of the membrane-linked protein A family [11], such as ANXA2, which promotes the formation of autophagosomes and mediates the fusion of autophagosomes and lysosomes [39, 40]. The role played by ANXA10 in the autophagic process warrants further investigation. 13. Gonzalo DH, Lai KK, Shadrach B, Goldblum JR, Bennett AE, Downs-Kelly E, et al. Gene expression proﬁling of serrated polyps identiﬁes annexin A10 as a marker of a sessile serrated adenoma/polyp. J Pathol. 2013;230:420–9. 14. Kim JH, Kim K-J, Rhee Y-Y, Bae JM, Cho N-Y, Lee HS, et al. Gastric-type expression signature in serrated pathway-associated colorectal tumors. Hum Pathol. 2015;46:643–56. 15. Sajanti SA, Väyrynen JP, Sirniö P, Klintrup K, Mäkelä J, Tuomisto A, et al. Annexin A10 is a marker for the serrated pathway of colorectal carcinoma. Virchows Arch. 2015;466:5–12. While our ﬁndings showed that the knockdown of ANXA10 induced ferroptosis and hindered the malignant proliferation and migration of CRC cells, it remains unclear whether the ferroptosis triggered by ANXA10 downregulation can enhance the sensitivity of existing chemotherapy, targeted therapy, or immunotherapy regimens. Therefore, it is necessary to conduct cellular and animal-level experiments to demonstrate the efﬁcacy of ANXA10 knockdown in combination with other therapeutic regimens. In addition, further studies should focus on the mechanisms by which ANXA10 affects TFRC degradation in lysosomes and understand how ANXA10 regulates the autophagy cycle. 16. Kim JH, Rhee Y-Y, Kim K-J, Cho N-Y, Lee HS, Kang GH. Annexin A10 expression correlates with serrated pathway features in colorectal carcinoma with micro- satellite instability. APMIS. 2014;122:1187–95. 17. Cell viability and migration were rescued by Ferrostatin-1 in ANXA10 knockdown colorectal cancer cells inhibiting autophagy-mediated TFRC degradation, thereby sup- pressing cell survival and migration ability. Our results demon- strated that treatment with Fer-1 restored the decreased cell viability of HCT116 and HT29 cells caused by ANXA10 knockdown (Fig. 6A, D), while having no signiﬁcant effect on the survival of control cells. Fer-1 greatly improved the migration ability of ANXA10 knockdown HCT116 (Fig. 6B, C) and HT29 cells (Fig. 6E, F). These ﬁndings collectively showed that the ferroptosis inhibitor recovered the survival and migration capacity of knockdown ANXA10 CRC cells. Cell Death and Disease (2023) 14:588 X. Wang et al. 11 DISCUSSION ANXA10, a calcium-dependent phospholipid-binding protein, has been linked to poor survival in CRC [33] and an enhanced risk of developing heterochronic serrated lesions in individuals with serrated lesions [12]. This study is the ﬁrst to investigate the expression and mechanisms of ANXA10 in the serrated carcinoma pathway of CRC. Consistent with previous studies, ANXA10 is speciﬁcally highly expressed in SSL and BRAFmut CRC and is associated with poor prognosis. single-cell sequencing and immunohistochemical assays conﬁrmed that ANXA10 was predominantly expressed in epithelial cells with serrated lesions. The datasets generated during the current study are available in the NIH National Center for Biotechnology Information Sequence Read Archive (SRA) repository, http:// gy q p y p www.ncbi.nlm.nih.gov/bioproject/950129 and Supplementary Table S3. www.ncbi.nlm.nih.gov/bioproject/950129 and Supplementary Table S3. FUNDING 27. Feng H, Schorpp K, Jin J, Yozwiak CE, Hoffstrom BG, Decker AM, et al. Transferrin receptor is a speciﬁc ferroptosis marker. Cell Rep. 2020;30:3411–23.e7. This work was supported by the Health and Family Planning Research Project of the Shanghai Pudong New Area Health Commission (PW2020D-12). 28. Montalbetti N, Simonin A, Kovacs G, Hediger MA. Mammalian iron transporters: families SLC11 and SLC40. Mol Asp Med. 2013;34:270–87. 28. Montalbetti N, Simonin A, Kovacs G, Hediger MA. Mammal families SLC11 and SLC40. Mol Asp Med. 2013;34:270–87. 29. Zhao N, Zhang A-S, Wortham AM, Jue S, Knutson MD, Enns CA. The tumor suppressor, P53, decreases the metal transporter, ZIP14. Nutrients. 2017;9:1335. AUTHOR CONTRIBUTIONS serrated neoplasia pathway toward colorectal cancer. Cell Mol Gastroenterol Hepatol. 2023;15:393–424. XBL conceptualized and supervised the study; XYW and YJZ collected the samples and analyzed the data; XYW and LJN performed the investigation and experiments; XYW and JNC drafted the manuscript; XBL and HMC reviewed and edited the manuscript. 25. Tang D, Chen X, Kang R, Kroemer G. Ferroptosis: molecular mechanisms and health implications. Cell Res. 2021;31:107–25. 26. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012;149:1060–72. REFERENCES Zhang X, Hu Z, Wang X, Li L, Zhu B, Lin X, et al. ANXA10 promotes melanoma metastasis by suppressing E3 ligase TRIM41-directed PKD1 degradation. Cancer Lett. 2021;519:237–49. 18. Sun R, Liu Z, Qiu B, Chen T, Li Z, Zhang X, et al. Annexin10 promotes extrahepatic cholangiocarcinoma metastasis by facilitating EMT via PLA2G4A/PGE2/STAT3 pathway. EBioMedicine. 2019;47:142–55. 19. Kodaira H, Koma Y-I, Hosono M, Higashino N, Suemune K, Nishio M, et al. ANXA10 induction by interaction with tumor-associated macrophages promotes the growth of esophageal squamous cell carcinoma. Pathol Int. 2019;69:135–47. 20. Shimizu T, Kasamatsu A, Yamamoto A, Koike K, Ishige S, Takatori H, et al. Annexin A10 in human oral cancer: biomarker for tumoral growth via G1/S transition by targeting MAPK signaling pathways. PLoS ONE. 2012;7:e45510. This study provides strong evidence that ANXA10 is signiﬁcantly upregulated in patients with SSL and BRAF mutant CRC, and is strongly associated with poor prognosis. Our results indicate that knockdown of ANAX10 can inhibit the malignant capacity of colorectal cancer cells by inhibiting autophagy-mediated degra- dation of TFRC, leading to cellular ferroptosis. 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Phipps O, Brookes MJ, Al-Hassi HO. Iron deﬁciency, immunology, and colorectal cancer. Nutr Rev. 2021;79:88–97. 39. Morozova K, Sridhar S, Zolla V, Clement CC, Scharf B, Verzani Z, et al. Annexin A2 promotes phagophore assembly by enhancing Atg16L+ vesicle biogenesis and homotypic fusion. Nat Commun. 2015;6:5856. 40. Bustos V, Pulina MV, Bispo A, Lam A, Flajolet M, Gorelick FS, et al. Phosphorylated presenilin 1 decreases β-amyloid by facilitating autophagosome-lysosome fusion. Proc Natl Acad Sci USA. 2017;114:7148–53. ACKNOWLEDGEMENTS We are grateful to all the individuals who participated in the study. COMPETING INTERESTS The authors declare no competing interests. 30. Pinilla-Tenas JJ, Sparkman BK, Shawki A, Illing AC, Mitchell CJ, Zhao N, et al. Zip14 is a complex broad-scope metal-ion transporter whose functional properties support roles in the cellular uptake of zinc and nontransferrin-bound iron. Am J Physiol Cell Physiol. 2011;301:C862–71. The authors declare no competing interests. ETHICS APPROVAL AND CONSENT TO PARTICIPATE This study was approved by the Ethics Committee of Shanghai Renji Hospital. All study participants provided informed written consent before study enrollment. ETHICS APPROVAL AND CONSENT TO PARTICIPATE This study was approved by the Ethics Committee of Shanghai Renji Hospital. All study participants provided informed written consent before study enrollment. 31. Xiong Q, Li X, Li W, Chen G, Xiao H, Li P, et al. WDR45 mutation impairs the autophagic degradation of transferrin receptor and promotes ferroptosis. Front Mol Biosci. 2021;8:645831. 32. Shan J, Jiang W, Chang J, Zhou T, Chen Y, Zhang Y, et al. NUF2 drives cho- langiocarcinoma progression and migration via inhibiting autophagic degrada- tion of TFR1. Int J Biol Sci. 2023;19:1336–51. © The Author(s) 2023, corrected publication 2023 We are grateful to all the individuals who participated in the study. ADDITIONAL INFORMATION Supplementary information The online version contains supplementary material available at https://doi.org/10.1038/s41419-023-06114-2. 33. Bae JM, Kim JH, Rhee Y-Y, Cho N-Y, Kim T-Y, Kang GH. Annexin A10 expression in colorectal cancers with emphasis on the serrated neoplasia pathway. World J Gastroenterol. 2015;21:9749–57. Correspondence and requests for materials should be addressed to Xiaobo Li. 34. Ma M-Z, Chen G, Wang P, Lu W-H, Zhu C-F, Song M, et al. Xc- inhibitor sulfasa- lazine sensitizes colorectal cancer to cisplatin by a GSH-dependent mechanism. Cancer Lett. 2015;368:88–96. Reprints and permission information is available at http://www.nature.com/ reprints Cell Death and Disease (2023) 14:588	37,805
tel-04448819-2023-GMADA-HENDA-10fce70f-VO.txt_17	French-Science-Pile	Open Science	Various open science	null	Des expériences sοcialisatrices à la pratique entrepreneuriale des femmes migrantes d'οrigine tunisienne : une apprοche par la cartοgraphie sensible. Gestion et management. Normandie Université, 2023. Français. ⟨NNT : 2023NORMR080⟩. ⟨tel-04448819⟩	None	French	Spoken	7,987	11,798	Ce mode d’entrée en entrepreneuriat pourrait être schématisé ainsi : 435 Figure 72: Vers une appréhension d’un entrepreneuriat stimulation L’indifférence de la mère Le partenariat avec le mari L’impact douleureux de l’agresseur Patrimoine disposition nel contexte présent •Mariage d'amour •Complicité du couple •Confiance mutuelle •A l'hypersensibilité •Au leadership •Au care •Migration inversée subie Entrepreneuriat stimulation •mariage avec soi même •équilibre familialprofessionnel •plus value renouvellée Patrimoine religieux en ressourcement Depuis 2019 entrepreneure en France de1983 à 1986 en Tunisie puis retour en France Age adulte en France 2008 : Mariage en France 436 Conclusion du chapitre L’objectif de ce dernier chapitre de la partie des résultats a été de porter notre appréhension de la pratique entrepreneuriale en analysant l’association des dispositions intériorisées suite aux expériences passées, à un contexte présent et à un jeu d’acteurs influents. Cette analyse a été rendue possible à travers l’analyse des portraits des femmes entrepreneures migrantes d’origine Tunisienne. Notre analyse de ces portraits nous a amenées à identifer, dans un premier temps, les dispositions de nos répondantes, en tant que manières d’agir et de réfléchir se transférant dans plusieurs contextes de leurs parcours. Il s’agit, à la fois, de dispositions générales s’actualisant d’une étape à une autre de leurs vies, mais aussi de propensions contradictoires mettant en évidence des crises dispositionnelles. Dans un second temps, nous avons mis en relief les contextes présents de nos informantes en explicitant leurs prises de décisions et leurs différentes mises en œuvre. Ceci nous a aidées à mieux comprendre leur situation actuelle. Nous nous sommes focalisées sur deux décisions importantes, nous semble-t-il, à savoir, le mariage et la migration. Au final, il nous a été possible d’appréhender la pratique entrepreneuriale de ces femmes migrantes tout en mettre en évidence des modes d’entrée en entrepreneuriat. Ceci a été rendu possible à travers une typologie associant les variations individuelles dans leur finesse, mais aussi les grands marqueurs structurels de notre terrain de recherche. Cinq modes d’entrée en entrepreneuriat féminin dans le contexte migratoire ont pu être identifiés afin de répondre à notre problématique. Il s’agit de : - L’entrepreneuriat réparation. - L’entrepreneuriat épanouissement. - L’entrepreneuriat stimulation. - L’entrepreneuriat délivrance. - L’entrepreneuriat care. Conclusion de la partie II Cette thèse s’est organisée autour de trois grandes parties. Dans un premier temps, nous avons présenté dans la partie I, les clés de lecture de ce travail doctoral. Nous y avons exposé le contexte de cette thèse qui maillait le marché, l’entrepreneuriat féminin et la migration, le cadre théorique mais aussi les dispositifs méthodologiques. Puis, dans un second temps, nous avons explicité, dans la partie II nos résultats de recherche tout en construisant les portraits sociologiques adaptés aux cas des cinq femmes entrepreneures migrantes, en mobilisant « le cross mapping » notre nouvelle méthode cartographique en marketing. Ensuite, nous avons tenté d’analyser ces portraits en mettant en lumière à la fois, les dispositions de nos répondantes, la manière dont est construit leur contexte présent ainsi que les jeux de rôles et de pouvoir se jouant entre les principaux acteurs de leurs parcours de vie. Nous avons alors été capables de capter des manières d’agir et de réfléchir de ces femmes résultantes des effets de leurs expériences socialisatrices passées et des éléments de leurs contextes présents. Dans cette perspective, il nous a été possible de détecter comment elles ont pu réaliser leur potentiel entrepreneurial sous des modes d’entrée différents. A présent, lors de la troisième partie de ce travail doctoral, nous allons mettre en lumière l’apport de cette thèse. Ceci prenra forme à travers la présentation de notre apport méthodologique au regard d’un terrain particulier et sensible, mais aussi en discutant la perspective dispositionnelle et contextuelle, la perspective intersectionnelle et positionnelle en entrepreneuriat féminin migrant et la perspective critique en Marketing. 438 Partie III : Discussion 439 Introduction de la Troisième Partie La thèse se poursuit par cette partie consacrée à la mise en perspective de nos résultats par rapport à la littérature. Cette dernière nous a servi de contextes croisés pour appréhender la manière dont les femmes entrepreneures migrantes se sont construites socialement et psychologiquement dans le contexte entrepreneurial en intégrant le marché. Pour pouvoir mesurer l’apport de cette thèse, nous avons fait le choix d’organiser cette troisième partie comme suit : - Discuter la réflexivité du chercheur dans cette thèse, de son immersion dans la recherche, de son évolution après la recherche, pour identifier les caractéristiques d’une recherche « care » à partir de l’expérience singulière de ce travail doctoral. - Discuter la spécificité des modalités de socialisation, identifiées dans cette thèse, avec la perspective dispositionnaliste et contextualiste de Lahire (2005). - Discuter les particularités des modes d’entrée en entrepreneuriat identifiés dans ce travail doctoral avec les éléments théoriques d’influence du processus de la réalisation du potentiel entrepreneurial féminin en contexte migratoire. - Discuter les modalités d’intégration du marché et des différentes dimensions identifiées dans nos résultats, avec une perspective critique en marketing. Cette dernière partie sera présentée en deux grands chapitres : le premier est consacré à la présentation de l’apport méthodologique du cross mapping et à la réflexivité du chercheur qui prend soin du terrain. Quant au second chapitre, il se focalise sur la discussion du social individualisé détecté chez nos répondantes mais aussi sur la discussion de la construction sociale d’une posture légitime de la femme entrepreneure sur le marché. 440 Chapitre VI : Le cross mapping : de l’apport méthodologique à la recherche « Care » Introduction du chapitre L’objectif de ce chapitre est de mettre en lumière la contribution méthodologique de ce travail doctoral qui s’est proposé de construire le dispositif cross mapping. Celui-ci a servi la reconstruction du parcours de vie de nos répondantes, par la co-création de cartes dessinées et figurées, suivie d’une discussion de groupe. Le but était de rendre possible la verbalisation bloquée sur un terrain sensible. Inspiré en partie par la cartographie sensible, le cross mapping bénéficie de l’apport théorique fort de la cartographie. Elle est en fait une méthodologie particulièrement mieux adaptée que les méthodes projectives classiques et les méthodes biographiques narratives, jusqu’alors utilisées. Ce dispositif bricolé permet d’avoir des données plus riches et plus fiables, tout en rétablissant une relation de recherche asymétrique entre le chercheur et son terrain. En outre, la mise en place du cross mapping en tant que méthode appartenant au registre sensible, permet également le déploiement d’une pratique réflexive laissant la place à une forme de discussion interne du chercheur et encourageant une forme de bienveillance avec son terrain, dans une relation considérée comme care. Nous organisons ce chapitre en deux sections dédiées respectivement à la contribution technique de la méthode et à la réflexivité du chercheur. Figure 73: Construction du chapitre VI le cross mapping de l apport mét 1. Le cross mapping : vers une contribution méthodologique sensible en marketing Lorsque nous avons été bloquées dans notre recherche par les difficultés d’expression et de verbalisation de nos répondantes, nous avons tenté de trouver une solution en mobilisant des dispositifs cartographiques pour aider au rééquilibrage de la relation et aider ainsi l’expression des répondantes. Par ailleurs, en organisant des réunions de groupe, nous évitions la frustration du face à face et nous encouragieons une expression étayée par le groupe. Notre bricolage méthodologique le cross mapping a, en effet, permis de combler les lacunes des méthodes discursives et biographiques classiques que nous avions mobilisées dans un premier temps. Nous présenterons tout d’abord ce que la cartographie sensible, en tant que cadre théorique, apporte à ce genre de terrains, puis nous discuterons les contributions de notre protocole méthodologique. 1.1. La cartographie sensible : les contributions du cadre théorique De prime abord, la construction cartographique sensible renvoie à une posture épistémologique compréhensive et interprétative. De plus, elle semble permettre également une posture constructiviste entre autres dans le cadre de la recherche intervention (Gaujal, 2016, De Robert et al, 2017 ; Paquin et Noury, 2018 ; Barelle, 2020). En effet, grâce à cette forme de cartographie, les géographes ont pu interpréter les motivations sous-jacentes des déplacements des personnes et de leurs mobilités dans les espaces géographiques donnés (Mekdjian, 2016). Ce positionnement entre la théorie et la pratique a donné lieu, par exemple, à une meilleure compréhension des comportements de consommation de l’eau potable dans des zones rurales africaines (Lefebvre et La gestion plus efficace des ressources naturelles dans les milieux ruraux a alors été rendue possible grâce à la construction d’une carte sensible des utilisateurs d’eaux. Dans ce cas précis, la cartographie sensible a été rendue possible par la mobilisation des savoir-faire artisanaux des populations interrogées pour mieux comprendre l’espace étudié (par exemple la broderie sur tissu, la poterie, les petits tapis brodés au fil, les tapis en paille...). 443 La cartographie sensible en géographie a aussi permis d’intervenir dans le domaine agricole pour concevoir un modèle de cultures co-construit avec les populations des habitants du terroir de Ziguéna au Mali (Traoré et Le Bars, 2018). En effet, la représentation de la dynamique agricole en zone cotonnière co-construite avec les popultations du village, a tenu compte des choix des cultures, du type de sol et de la rotation culturale. Les résultats issus de la cartographie participative sensible et du jeu d’acteurs, ont permis d’établir une carte d’occupation finale du terroir tout en tenant compte des ressources locales. En mobilisant cette méthode créative pour étudier plus particulièrement les populations marginalisées (par exemple analphabètes), il s’est avéré que les ressentis d’injustice représentés dans les cartes sensibles produites ont contribué à développer les recherches questionnant les changements sociaux, du point de vue des acteurs (Peluso, 1995). Pour les géographes, les apports de la cartographie sensible, s’avèrent, par conséquent, considérables dans la mesure où ce schéma participatif permet d’aboutir à des solutions adaptées aux populations étudiées, la solution étant proposée par les acteurs du territoire et non par les chercheurs-experts. Pour les géographes, la reconnaissance de l’expertise des répondants sur leur propre territoire, rééquilibre la relation de recherche (Burini, 2012, Olmedo, 2011 et 2015, Barella, 2020). La cartographie sensible a permis aux géographes de reconnaitre même chez les répondants marginalisés, une forte capacité à parler pour eux-mêmes (Spivak, 1988). C’est ainsi que la méthode est utilisée également auprès des élèves dans une perspective pédagogique (Gaujal, 2017, 2019). Les géographes passent ainsi, à travers la cartographie sensible, d’une recherche etic qui permet une vue d’ensemble de la culture, sur la base de critères objectifs de catégorisation des populations, à une recherche emic, qui étudie la culture de l’intérieur dans le cadre d’une approche ethnographique dynamique (Zouaghi, 2012). Les contributions épistémologiques de la cartographie sensible en géographie peuvent être récapitulées dans la figure suivante : 444 Figure 74: les contributions épistémologiques de la cartographie sensible en géographie Cartographie sensible recherche symétrique Intervention Interprétation Compréhension Construction Malgré le rétablissement de l’asymétrie des rapports et de l’accès au savoir spatial des populations, la cartographie sensible ne semble pas effacer totalement la frustration du face à face, notamment dans un contexte de cultures communautaires ou dans le cas d’objet de recherche douloureux (Gagnon et al 2019). C’est la raison pour laquelle nous avons réfléchi à un bricolage plus adapté dans le cadre du dispositif du cross mapping qui tienne compte de ces blocages. Nous tenterons dans la sous-section suivante de montrer les contributions qui ont été constatées lors de la mise en place de notre méthode, qui s’avèrent plus larges que celles présentées par les géographes. 1.1. Le cross mapping : vers un apport méthodologique Le cross mapping est avant tout une méthode dont la mobilisation se propose de débloquer des situations communicationnelles compliquées par une relecture à la fois du positionnement du chercheur, en charge de l’enquête, mais aussi de l’enquêté généralement en situation de vulnérabilité sur un terrain sensible. Cette dualité qui jalonne l’objectif de mise en œuvre de la méthode est au cœur de son apport au regard de la sensibilité des personnes et des terrains. L’apport du cross mapping en marketing concerne à la fois le rétablissement de la symétrie chercheur-répondant, voire le renversement de la relation du pouvoir, mais aussi le transfert plus fluide et plus robuste (sincère) des données qualitatives sensibles. Le tableau comparatif ci-dessous récapitule les contributions de la méthode : 445 Méthodes projectives Méthodes biographiques Cross mapping narratives Contenu Attribution à une autre entité Expérience de consommation Entretiens bio graphiques libres facilités par la (personne ou chose) des qualités , réelle telle qu’elle a été vécue confection de la carte sensible et par l’entretien de sentiments, besoins, Données défauts, désirs, croyances, par l’informant / faits réels et groupe, en vue de reconstruire le parcours de vie tel motivations et description de la consommation qu’il a été vécu par le répondant, vers un éclairage par comportements qu’on méconnaît ou exemple des projets identitaires et des manières refuse en soi, en vue de solliciter d’agir, de réfléchir et de sentir. l’imaginaire du répondant et faciliter Grille de lecture des domaines des pratiques de son expression par rapport à une consommation à la discussion du répondant: école, situation ou à un sujet bien déterminé. famille, travail, sociabilités, culture, territoire... Entretiens oraux, visuels, de Entretiens oraux /données Entretiens oraux construits par la production personnification, des jeux de rôles, secondaires comme le dossier artisanale/artistique, manuelle co-créée avec le rêves éveillés dirigés. médical, des actes juridi ques, répondant. ou encore des témoignages de proche.s Formes Descriptive non structurée Narrative/descriptive 446 Artistique/ artisanale et narrative/descriptive Statut de l’informant Sujet /Acteur Acteur / informateur Co-acteur et agent multi déterminé par la variation des effets des expériences intériorisées Posture du chercheur Etic Etic/Emic Emic Subjective Objective/subjective Objectivé à partir des données subjectives synchronique Chronologique / spécifique au Diachronique et synchronique Statut de la connaissance produite Structure du récit récit Modalités et taille de l’échantillon * Echantillon de convenance / de Echantillon de convenance / Echantillon de convenance / de jugement jugement de jugement Un ou plusieurs répondants et saturation théorique Un ou plusieurs répondants jusqu’à *Un ou plusieurs répondants possible saturation jusqu’à saturation Tableau 21: Différences entre le cross mapping, les méthodes biographiques narratives et les méthodes projectives, Adapté de : Laplanche et Pontalis, (1967) ; Lombart, (2008) ; Özçağlar-Toulouse, N. (2008) ; Bah et al, (2015) ; Trizzulla (2018). 447 Comme le montre le tableau 22, même si le cross mapping peut se rapprocher des méthodes projectives et de la biographique narrative, il n’en reste pas moins que certaines caractéristiques lui sont spécifiques : - Il s’agit d’une reconstruction de tout le parcours de vie personnel, sinon d’une longue période, s’installant dans une chronologie biographique Cela le différencie des récits de vie, par exemple, qui concernent un ou des épisodes bien déterminés ou encore des méthodes projectives qui sont relatives à une situation bien déterminée ou à un sujet identifié (Laplanche et Pontalis, 1967 ; Lombart, 2008 ; Besson et Mendoza, 2019). - Il s’agit de la mobilisation de techniques cartographiques artisanales, de créations artistiques et/ou artisanales et de discussions individuelles (pendant la création de l’artéfact) puis en groupe (après la création) afin de faciliter l’expression verbale de l’informant. Le cross mapping se distingue alors par cette stratégie méthodologique plurielle : artistique et/ou artisanale, puis verbale, donnant accès aux parcours de vie tel que ressenti et vécu par l’individu. Cela le différencie des méthodes projectives qui cherchent principalement à révéler les éléments présents dans le subconscient ou l’inconscient des répondants (Lombart, 2008 ; Besson et Mendoza, 2019). - Le récit reconstruit suite à la mobilisation du cross mapping est structuré selon la grille de lecture des cadres socialisateurs basée sur une temporalité diachronique et synchronique permettant une approche duelle des évènements du parcours, (la reconstruction, à la fois, installée dans la chronologie mais aussi dans la situation à un instant déterminé) ; ce qui permet d’avoir par exemple, des portraits sociologiques complets et très singuliers. Les méthodes projectives, elles, se focalisent sur le contenu du subconscient/inconscient d’un sujet à un moment donné (Besson et Mendoza, 2019). Les méthodes biographiques narratives comme les récits de vie, suivent, quant à elles, la chronologie du récit sans prendre en compte la situation actuelle du sujet (Özçağlar-Toulouse, 2008 ; Bah et al, 2015). 448 - La posture du chercheur en cross mapping est emic grâce à l’immersion dans le monde du répondant tant dans le choix de la technique de création que sur sa prise en main de la narration. Cette posture contribue, à la fois, à la captation des effets des cadres socialisateurs, mais aussi, dans un second temps, à détecter le sens attribué aux pratiques individuelles. En revanche, le chercheur est dans une posture etic en méthodes projectives et peut accéder à une posture emic en méthodes biographiques. Pour catégoriser les contributions méthodologiques du cross mapping, nous avons fait le choix de les répartir en trois grandes classes : la richesse des données, la robustesse des informations et la relation de recherche plus symétrique. 1.1.1. Des données plus riches Le cross mapping se déploie sous la forme de trois ateliers qui visent à reconstruire tout un parcours de vie dont les données s’avèrent sensibles parce qu’elles touchent à l’intime ou au sacré de la personne, qui peut se sentir menacée par l’intrusion dans sa sphère (Brito et Pesce, 2015). Outre le fait que chercheur peut être conscient d’être intrus et menaçant, il peut ressentir de la frustration par rapport à la qualité des données recueillies dans ce genre de terrain. Avoir des données particulièrement superficielles crée, en effet un malaise chez le chercheur. Avec le cross maping, le contenu recueilli est particulièrement riche, d’abord parce que les entretiens biographiques reconstruisant tout le parcours de vie de l’informant, donnent à voir une quantité d’informatins importante. De plus, la structure même des ateliers met l’accent sur l’importance de certains évènements dans le parcours et ce à plusieurs reprises pour ne laisser échapper aucune information. Plus précisément, lors du premier atelier consistant à dessiner le parcours des répondantes (en y mettant des symboles libres associés à chaque évènement important), le répondant fera un premier jet de son histoire. Il reviendra ensuite sur chaque événement, lors du deuxième atelier et pourra approfondir certains faits à sa convenance. Enfin, la vérification / confirmation des données relatives à tout le parcours, sera faite. 449 Dans cette perspective, et à la suite de toutes ces étapes, le chercheur reconstruit tout le parcours de vie de l’informant qui raconte à la fois, en dessinant, en fabricant puis en discutant en groupe. Toutes ces étapes s’imbriquent et créent une sorte de mécanisme de recueil itératif de données dont le transfert se fait dans le cadre d’un processus temporel codifié et appuyé par les compétences artistiques, manuelles puis verbales de l’informant. La durée de 9 heures en moyenne passée avec chaque répondante de notre terrain sensible, peut témoigner de la richesse des données recueillies retraçant tout le parcours de vie de la date de naissance jusqu’à aujourd’hui, en passant par l’enfance, l’adolescence, la jeunesse jusqu’à l’âge adulte actuel, avec toutes les expériences vécues, les moments de douleur, de joie, de déception, etc... Par ailleurs, la particularité du protcole participatif du cross mapping, en ce qu’il implique le chercheur dans la production de la connaissance, incite le répondant à raconter plusieurs détails parfois très intimes et particulièrement douloureux. Cela est rendu possible, nous semble-t-il à partir du sentiment d’étayage. Ce ressenti est le résultat du réequilibrage de la relation et de l’ambiance apaisante et confortable qu’apporte le « faire avec les mains » qui nécessite un temps long. Les étapes nombreuses et les rendez-vous successifs permettent également de construire une relation de confiance où le rythme est donné par le répondant. A titre d’exemple, nous pouvons citer le cas d’Ambre, qui avait du mal à raconter un épisode douloureux d’agression sexuelle lors du premier atelier du dessin et a préféré juste mentionner quelques informations vagues autour de l’évènement. Mais petit à petit, elle s’est ouverte lors de l’atelier de production de l’artefact au moment du coloriage de son attrape-rêve. Elle a donné les détails de l’incident, le moment de l’agression, son âge à cette époque et les séquelles qui persistent jusqu’à aujourd’hui. Dans le même ordre d’idées, l’informante Kamélia, a eu des difficultés à se confier par rapport à son statut actuel de femme de ménage (en plus d’être entrepreneure). Elle s’est révélée graduellement lors du deuxième entretien du dessin du parcours de vie. La liberté des entretiens et des procédés de production de la connaissance dans le cross mapping étaye l’informant et apaise sa frustration puis débloque progressivement sa verbalisation. Des données riches dont le transfert devient particulièrement fluide voire naturel, y sont alors rendues possibles. 1.1.2. Des données plus fiables et plus crédibles Le cross mapping s’avère être un dispositif méthodologique fournissant une quantité particulièrement importante et riche d’informations relatives au parcours de vie. La méthode gagne également en capacité d’accès à des informations parfois refoulées par l’informant avant la collecte des données. Le recueil des données grâce au cross mapping donne aussi à voir des informations crédibles et fiables récoltées tout au long du processus. En recherche qualitative, on parle de qualité, de crédibilité ou de fiabilité des données, dès lors que les observations sont stables dans le temps (Kirk et Miller, 1986 ; Drucker-Godard et al, 2014). La transparence du processus de recueil des données et le recoupement des sources à différents moments, peuvent également apprécier la crédibilité ou la fiablilité des informations recueillies (Miles, Huberman et Saldana, 2013). En effet, les informations riches permettent des analyses objectivées de données subjectives (Trizulla, 2018), rendues possibles par la structure diachronique et synchronique du parcours raconté. Cette particularité rend les connaissances sur le répondant plus robustes, ce qui est souvent compliqué dans des terrains sensibles. Dans cette lignée d’idées, l’objectivation du subjectif se fera en deux temps. Premièrement, par l’adoption de la même structure des sphères sociales pour tous les répondants (par exemple, famille, école, travail, sociabilités...), lors du recueil des données dans les différents ateliers de production. Deuxièmement, par la détection des éléments de récurrence des éléments d’analyse (par exemple les dispositions) du contenu du parcours en fonction de leur transférabilité dans plusieurs contextes sociaux du parcours de vie de l’informant. Ceci représente une occasion supplémentaire de vérifier la robustesse et la crédibilité des informations. Le terrain sensible pourrait être paradoxalement attractif pour les chercheurs voulant explorer des situations riches. Néanmoins, faire de la recherche sur ces terrains renvoie à la fois, à plusieurs défis mais aussi à plusieurs risques dont le risque « compassionnel » du chercheur visà-vis de ses informants (Demazière et Zune, 2019). Ce risque peut parfois complexifier l’accès à des données fiables. En effet, les informants vul rables ou encore les données sensibles peuvent influencer le chercheur, qui par contre-transfert, peut les surinterpréter ou déformer leur interprétation. 451 Néanmoins, la capacité du cross mapping à capter l’effet des structures sociales génératrices se situant en-dehors de l'individu, comme la famille, l’école, le travail, les sociabilités, etc... rend possible la détection de la transférabilité des manières d’être et d’agir dans différents contextes et situations, à travers la dimension synchronique (relative à un instant précis) et la dimension diachronique (reconstruisant l’évolution de la pratique dans le temps). La structure du récit produit répond à cette dualité diachronique et synchronique qui enrichit les analyses. En effet, se focalisant sur l'évolution du phénomène étudié au fil du temps (par exemple la relation avec la mère) en retraçant son histoire et les transformations qu'il a subies, la caractéristique diachronique permet à la fois d’appréhender le passage d’un état à un autre mais aussi d'identifier les facteurs ayant influencé la transformation (Guillaume, 2010 ; Nielsen et al, 2021). En outre, l’approche synchronique se concentre, elle, sur un élément (par exemple le positionnement dans la famille) à un moment donné tout en décrivant ses caractéristiques, les relations qu’il met en œuvre ainsi que les structures qu’il déploie à ce moment bien déterminé. Ce faisant, l’approche synchronique ne se préoccupe pas de son évolution dans le temps (Ternoy, 1997 ; Teyssier et Denoux, 2013 ; Crahay, 2022). Le cross mapping met en pratique un protocole alternant ces deux approches, diachronique et synchronique, ce qui fournit toutes les caractéristiques nécessaires à une production de connaissance particulièrement riche et crédible parce qu’elle articule les deux perspectives afin d’avoir une compréhension complète du phénomène étudié. Par sa dualité synchronique et diachronique, le cross mapping est en accord avec les travaux de didier (1971). La capacité de la méthode à gagner en fiabilité et en crédibilité, est alors rendue possible. Ceci se déploie en pratique, à la fois par la succession des étapes de production (le dessin, l’artefact et la discussion en groupe), mais aussi par la structuration du contenu produit. En effet, le chercheur a, à plusieurs reprises, l’occasion de relancer la conversation en dessinant et en figurant le parcours et même en groupe, en cas de doute, de façon à être dans une compréhension au plus proche du sens voulu par l’informant. Par ailleurs, la perspective dispositionnelle contextualiste adoptée pour structurer les données est également à l’origine d’une vérification de la fiabilité des données. Plus précisément, la perspective dispositionnelle adoptée dans notre travail doctoral, nécessite de capter la 452 réccurence des manières de voir et d’agir dans plusieurs contextes du parcours, et favorise ainsi la consolidation des analyses. La grille de lecture structurée de la même façon, à travers les mêmes sphères sociales dans chaque histoire, pourtant singulière, favorise en effet, la vérification complémentaire de la fiabilité et la crédibilité des données. Le cross mapping met en pratique un protocole alternant les approches, diachronique et synchronique, ce qui fournit toutes les caractéristiques nécessaires à une production de connaissance particulièrement crédible parce qu’elle articule les deux perspectives afin de construire une compréhension complète du phénomène étudié. La fiabilité des données est également consolidée par la transparence du processus mis en place. En effet, la rédaction du protocole clair, des étapes détaillées du recueil des données, du contexte de la production dans les trois ateliers, appuyée par les notes du journal de bord, rend visibles des précautions mises en œuvre. 1.1.3. Une relation de recherche plus équilibrée Lorsque nous avons mené la première étape de ce travail doctoral dans sa partie exploratoire, nous avons été frappées par le degré d’asymétrie qui a pu s’installer sur le terrain rural tunisien. Cette asymétrie a dénaturé la situation d’enquête dans la mesure où les femmes intérrogées ont refusé de se livrer avec sincérité. Plus précisément, malgré les similitudes et les points de rapprochement entre le chercheur et son terrain (même origine, même genre, même langue...), un blocage communicationnel s’est tout de suite installé et les informantes ont pu ressentir un embarras palpable chez le chercheur. La parole était bloquée et il est apparu rapidement en discutant avec des spécialistes d’éthnopsychologie que ce blocage était dû à cette asymétrie. Force est de souligner que l’asymétrie renvoie à une situation où les deux parties engagent un rapport de nécessité déséquilibré. Une partie détient le savoir ou la compétence et une autre est censée souhaiter se dévoiler (Zouaghi, 2012 ; Charlier et Biémar, 2012 ; Boucenna, 2017). Qu’elle soit le résultat d’une comparaison à partir de soi, ou encore à partir d’autrui, l’asymétrie s’installe en cas d’absence de réciprocité (Zielinski, 2011). 453 En l’occurrence, afin d’avancer dans la recherche, il nous a été impératif de faire disparaitre cette asymétrie par un renversement radical du rapport de force en redonnant le pouvoir à l’informante (Zielinski, 2011), par le choix d’une activité qu’elle maitrise, ce qui n’est pas le cas du chercheur (Olmédo, 2015). Le recours au cross mapping avec les femmes entrepreneures en contexte migratoire français a été d’une efficacité considérable dans le rééquilibrage de la relation. Le déblocage communicationnel en a été la conséquence. Avec le protocole de production des données cartographiques sensibles, nous sommes passées d’une situation d’asymétrie, valorisant le chercheur en tant que sachant, à une situation de symétrie où chacune des parties posséde sa zone de pouvoir : le sujet de l’étude et la connaissance du dispositif par le chercheur et la connaissance de la pratique manuelle/artistique et son parcours de vie par l’informante. Ce rééquilibrage a eu lieu avec toutes les répondantes. Nous pouvons citer l’exemple d’Amira, dont le parcours de vie est particulièrement douloureux. Lors de notre premier rendez-vous, elle avait des difficultés à se livrer et a été peu loquace sur son parcours. Lors de notre deuxième entretien, nous avons repris le dessin du parcours de vie pour voir s’il manquait quelques détails et là, elle a commencé à parler de sa passion pour le design sur cake, pour enchainer avec des corrections sur le dessin afin qu’il soit bien réalisé sur la pâte à sucre (que nous avons créée ensemble lors du second atelier). Elle a graduellement donné plus de détails sur certains passages douloureux. Plus précisément, tout en nous apprenant à utiliser le matériel, à mélanger les couleurs, à coller les symboles avec de la colle naturelle, Amira a repris le pouvoir en communiquant son savoir-faire et a ainsi rééquilibré la relation. Ce processus a nécessité beaucoup de temps, ainsi les entretiens avec Amira ont duré de 3h00 à 3h45. De manière inattendue, un renversement de la situation d’asymétrie s’est aussi manifesté sous une autre forme. En effet, à la fin de l’atelier de discussion de groupe, Warda, notre informante styliste couturière, a proposé de nous faire faire le protocole du cross mapping, pour révéler notre propre parcours de vie. Sa proposition a suscité l’intérêt de toutes les autres répondantes qui l’ont encouragée. Cette proposition inattendue montre que les répondantes se sont senties autorisées à prendre en main la suite du processus relationnel et à faire vivre au chercheur ce qu’elles ont vécu. A ce moment là, elles se sentaient autorisées à être les cheffes d’orchestre, elles détenaient le pouvoir. 454 Ces analyses montrent que le cross mapping, la méthode cartgraphique sensible, est susceptible de contribuer à un transfert de données plus riches, plus profondes et plus fiables dans le cadre d’une relation de recherche plus équilibrée. Comme nous l’avons montré, le vécu relationnel à travers le cross mapping a été intense avec les répondantes. Il a, en effet conduit « un dialogue interne » que nous analysons ci-dessous dans la perspective des théories du care. 455 2. De la réflexivité du chercheur à la recherche care Pour le chercheur en sciences sociales, la réflexivité est un exercice essentiel en vue de rendre compte du côté humain de la recherche. Pour mener une pratique réflexive, le chercheur doit articuler deux niveaux : la démarche et la finalité de la production de la connaissance (Blanchet, 2009). Nous présentons dans cette section notre pratique réflexive sur ce travail doctoral en mettant en lumière la manière dont nous nous sommes intérrogées à la fois à propos de la démarche mais aussi à propos des résultats de cette recherche. Cela nous a aidées à assumer notre subjectivité dans cette recherche participative pour construire une relation de confiance en recherche sensible. Cette décision d’assumer notre subjectivité nous a permis de passer de la question de la vulnérabilité celle du care. Les deux premières parties abordent la question de la reflexivité lors de la collecte des données, et la troisième porte sur l’analyse des résulats. Il est nécessaire de souligner que ces axes se croisent souvent, car la gestion de vulnérabilité appartient aussi bien à la démarche qu’à la construction des résultats. 1.2.Assumer la subjectivité en recherche participative Avant de commencer le terrain avec les femmes entrepreneures migrantes, nous avons effectué un travail d’introspection profond afin de prendre conscience de l’influence potentielle de notre propre identité et de nos croyances sur l’objet de la recherche. En effet, nous étions convaincues que la recherche serait participative avec un protocole méthodologique lourd et impliquant. Il nous a été par conséquent nécessaire de nous auto-analyser avant de nous lancer (Béji-Bécheur, Herbert et Özçağlar-Toulouse, 2011 ; Béji-Bécheur, Özçağlar-Toulouse et Zouaghi, 2012). S’auto-analyser revient à faire l’analyse de soi-même pour être conscientes de nos croyances, émotions et sensibilités. Cela est d’autant plus important que le chercheur risque de partager plusieurs points communs avec le terrain : l’origine, la langue, les traditions, l’idéologie, etc... Il a donc fallu anticiper un degré d’implication et de subjectivité inévitable dans ce genre de recherche. Par la suite, nous sommes passées à une étape cruciale, qui consistait à accepter les 456 risques et les défis liés à une recherche menée auprès de personnes fragilisées et vulnérables (Zielinski, 2011). Tout d’abord, se préparer à notre terrain a consisté à mener un dialogue interne pour une prise de conscience de nos préjugés et de nos doutes. Notre journal de bord a été un outil d’une aide précieuse pour la prise de conscience et pour pouvoir avancer sans blocages. Au fil des entretiens, nous avons tenu notre journal de bord en y notant des éléments descriptifs, méthodologiques et théoriques (Deslauriers, 1991), ainsi que des notes réflexives visant à faire le point sur nos émotions et nos ressentis anticipés et vécus (Baribeau, 2005 ; Gagnon et al, 2019). L’analyse des notes du journal de bord nous a données la possibilité de mettre en lumière les enjeux épistémologiques et méthodologiques propres à notre recherche, non seulement par rapport aux précautions à émettre lors des entretiens avec les répondantes, mais aussi aux investissements réflexifs à mettre en oeuvre (Baribeau, 2005). Photographie 44: Extrait du journal de bord introspectif avant le terrain Notre pratique réflexive sur nos motivations et nos intérêts personnels nous a effectivement permis de mettre en évidence notre projet individuel dans un premier temps. Ensuite, dans un second temps, elle nous a aidées à effectuer des mises au jour de notre liste de motivations afin de repositionner la recherche et nous rappeler sa finalité (Blanchet, 2009). 457 Plus précisément, réussir notre projet personnel passait par l’exploration riche du terrain afin de pouvoir construire notre thèse. Par conséquent, nous devions éviter les blocages qui ont eu lieu lors de la phase exploratoire. Dans cette perspective, nous avons adopté une position basse dans la mise en œuvre de la méthodologie, afin de favoriser un transfert fluide des données. Tout en prenant conscience de nos compétences et nos connaissances, suite à la pratique réflexive, nous avons réussi à mettre de côté nos croyances limitantes (liées à la culture d’appartenance notamment), pour nous ouvrir à la découverte des femmes interrogées, de leur vécu, sans parti pris. Cet exercice réflexif a donné lieu à une mise en place adaptée de la méthode participative en congruence avec le positionnement constructiviste de notre travail doctoral. L’annonce, avant chaque atelier, du caractère libre et créatif des activités a eu pour objectif de favoriser l’atmosphère apaisante de la recherche. La peur des silences et du refus de se livrer, notée dans notre journal de bord, a pu être travaillée mentalement et a conduit à accepter le rythme de la répondante avec patience et bienveillance. Nous avions été bien préparées à cet aspect sensible du terrain grâce aux notes du journal de bord (notes de vigilance mises à jour suite à chaque entretien). Ces notes ont servi considérablement tout au long des entretiens successifs. Par exemple, les notes du terrain avec Amira nous ont beaucoup aidées à avoir le comportement adapté lors du recueil des informations douloureuses d’Ambre, dans ses silences et son repli. En effet, une fois, le moment de silence respecté, la répondante s’est ressaisie, et a repris le dessin et la verbalisation. C’est ce que nos notes des terrains antérieures nous ont fait savoir et appris. Par ailleurs, l’exploration de notre réflexivité nous a aidées à accepter notre subjectivité, les expressions qui pouvaient en découler et d’ajuster nos comportements. Plus précisément, il a été question pour nous de réfléchir à la place et au rôle que nous occupions sur le terrain. Nous devions être dans l’enquête, en faire partie et co-créer l’expérience de la recherche avec nos répondantes. Nous devrions accepter à la fois la position basse d’apprentissage pour rééquilibrer la relation, mais aussi guider la recherche. Cela conduisait parfois à laisser une certaine empreinte personnelle dans le dispositif car notre participation était occasionnellement importante notamment lors des ateliers de co-création artisanale et/artistique. 458 Non sans charge mentale et émotionnelle - nous y reviendrons dans les paragraphes suivants l’exploration de notre réflexivité nous a permis d’assumer cette participation active sur le terrain. Photographie 45: Accepter la subjectivité par l’exploration de la réflexivité Tout en assumant graduellement notre subjectivité, nous avons alors peu à peu appris à gérer la participation dans la construction du matériau empirique en aidant les femmes entrepreneures à créer leurs artefacts et en dessinant avec elles leurs parcours de vie émotionnels. Grâce à l’exercice réflexif, nous avons pu faire la différence entre la participation dans la mise en place de la méthode et l’influence de l’expression du vécu de ces femmes. Plus précisément, la pratique réflexive a donné à voir une robustesse à la fois, des modalités de mise en pratique cartographique du dispositif méthodologique avec les informantes, mais aussi du transfert des données conformes aux parcours de vie de nos répondantes sans influence de notre part. Notre participation s’est articulée à la fois autour de la reconnaissance du savoir expérientiel des répondantes, d’une relation chercheur-informante plus égalitaire, mais aussi de la visée de 459 la conscientisation des parties participantes (Lewin, 1946 ; Ozanne et Saatgioclu, 2008 ; Dubost, 2018). Nous avons été fidèles à notre responsabilité par rapport à des terrains sensibles, où la parole des personnes vulnérables est particulièrement précieuse au regard de leur marginalisation. Nous avons été particulièrement vigilantes sur ce point. Nous mesurons, aujourd’hui que le processus réflexif a considérablement contribué à mettre en place une relation de qualité entre le chercheur et les répondantes, donnant lieu à une bonne qualité du recueil de données sur un terrain sensible (Dubost, 2018). En effet, après le dernier atelier de discussion en groupe, nous avons eu des retours fort intéressants des informantes se focalisant surtout sur : - L’apprentissage et l’échange avec l’autre, qui font du bien. - L’effet apaisant de reconstruire avec l’autre son parcours, avec ses joies et ses douleurs, tout en le représentant par le dessin et l’artefact. - La relation de recherche qui s’est installée sur le respect de l’intime et du sacré des répondantes. - La réflexion par rapport au rôle joué par ces femmes dans la société du pays d’accueil. - Le regard vers un futur prometteur en tant qu’entrepreneure migrante. Nous avons alors été ravies que la recherche ait pu mettre en évidence à la fois une richesse et une crédibilité de résultats mais également des bénéfices psychologiques pour nos répondantes (Dubost, 2018). Photographie 46: extrait du journal de bord : le retour des répondantes En somme, notre exercice réflexif a permis à la fois, une mise en lumière des sources potentielles de blocages sur le terrain sensible des femmes migrantes mais aussi une mise en garde contre les glissements vers une relation asymétrique. De plus, il a éclairé des compétences et des connaissances du chercheur pouvant y contribuer, ainsi qu’une initiation à sa participation fluide. Le tout en assumant sa subjectivité particulièrement cadrée par les objectifs de la recherche. En participant à la recherche, nous avons permis à nos répondantes de parler de façon approfondie de leur parcours en détaillant à la fois leurs expériences mais aussi les éléments de leur contexte actuel. Cette expression a été plus intime (Falchetti et al., 2016), grâce à la qualité relationnelle qui s’est installée entre le chercheur et ses informantes et à favoriser la construction d’un lien de confiance, essentiel dans ce genre de situation de co-création du savoir (Nelson et al., 1998 ; Mason et Pavia, 2006 ; Nau et al., 2016 ; Dubost, 2018). 1.3.Construire la relation de confiance avec le terrain Si la pratique réflexive a structuré l’avancement de notre recherche sur le terrain sensible des femmes migrantes, elle a été particulièrement prégnante dans l’établissement graduel du lien de confiance avec le terrain. Nous mesurons ici l’importance de ce lien dans le processus d’accès aux vies de nos répondantes. Nous pouvons même assurer qu’il a été question d’un privilège que nous avons négocié prudemment avec elles (Chesler, 1991), tout en se référant à nos notes réflexives qui orientaient tout le processus. La construction de notre relation de confiance avec notre terrain s’est déployée en trois phases qui ont été en congruence avec l’avancement de la recherche. En effet, nous avons été convaincues qu’il fallait un processus particulier de mise en œuvre afin d’introduire notre lien, le maintenir puis le consolider, comme nous le développerons ci-dessous. 1.3.1. L’introduction à la confiance Nous avons été extrêmement vigilantes avant et pendant cette étape. C’est la raison pour laquelle nous avons essayé de recueillir toutes les informations nécessaires. La préparation a 461 commencé avec la personne relais avec qui nous avons eu de longues discussions pour comprendre le contexte vécu par ces femmes migrantes. Nos préparations se sont focalisées sur deux axes. Tout d’abord, faire doucement passer le message qu’il s’agit d’une recherche, sans intimider mais en donnant suffisamment d’explications qui respecte l’informante et sa vulnérabilité. Ensuite, réussir à capter son attention pour préserver son intérêt pour les étapes suivantes. Nous devions par conséquent doser notre enthousiasme, ce qui n’était pas facile au regard de notre implication émotionnelle, sans pour autant tomber dans la froideur du premier contact. Nous savions que ce dernier est culturellement refoulé chez les populations tunisiennes extrêmement émotives et sociables et pour lesquelles le langage a une dimension affective importante (Ben Fadhel, 2010). Pour ce faire, nous avons dû puiser dans notre patrimoine culturel et intellectuel pour établir et préserver une relation de confiance avec les répondantes. Photographie 47: Extrait du journal de bord après la phase d’introduction Une fois la phase introductive s’est bien passée, le lien de confiance commençait à prendre forme d’une façon graduelle lors des rendez-vous suivants (de production des dessins puis des artéfacts.) 462 1.3.2. Le renforcement de la confiance Force est de constater que la production participative a naturellement permis une collaboration apaisante. Cette dernière s’est renforcée par la mise en place, de la position basse d’apprentissage du chercheur en reconnaissance du savoir-faire de la répondante, mais aussi de la capacité d’écoute à laquelle nous nous sommes initiées par l’exercice réflexif. En conséquence, le lien de confiance, préalablement installé lors de la phase introductive, a été renforcé de façon fluide. A titre d’exemple, nous pouvons mentionner le lien de confiance avec Kamélia qui a préféré nous léguer la mission du dessin, probablement parce qu’elle ne voulait pas affronter les évènements de sa vie. Nous avons été à l’écoute de sa demande tout en acceptant de jouer un double rôle : l’apprentie et la secrétaire. Kamélia nous a été très reconnaissante et a même mentionné son ressenti positif par rapport à ce mode opératoire particulier, lors de la discussion en groupe. L’exemple d’Ambre est également marquant au sujet de la construction et du maintien du lien de confiance. En effet, elle a relaté des épisodes très douloureux de sa vie que les membres de sa famille ignorent. Par ailleurs, tout au long de la démarche empirique, nous avons mesuré cette confiance qui prenait petit à petit corps au fil des rendez vous. Par exemple, nous avons été accueillies chez Warda et nous avons mangé ensemble, ce qui n’est pas commun entre Tunisiennes qui viennent de faire connaissance. De plus, elle nous a montré les endroits qu’elle aimait chez elle, comment elle les a décorés, quels souvenirs cela évoque pour elle... Nous avons également eu l’occasion de comprendre ses difficultés à avoir des enfants dès le deuxième rendez vous. L’exemple de Syrine la spécialiste de la résine, est aussi très illustratif. Avec elle, cela a été rapide dès le premier entretien. En effet, en raison de sa fragilité, elle a eu du mal à se livrer. Toutefois, quand elle a commencé à dessiner, elle nous a tout de suite dit qu’elle avait besoin de se livrer mais qu’elle avait toujours eu du mal à le faire. Elle a précisé qu’elle nous faisait confiance, et a commencé à dessiner tout en se livrant petit à petit pour raconter des scènes familiales douloureuses. Lors de cette partie de production, les échanges ont été très riches et les informantes ont ressenti le grand respect qui leur était voué. Il s’agissait d’autant de moments propices à renforcer la confiance. La confiance de l’après Au regard de l’objet de recherche, des populations cibles et du degré d’implication du chercheur, la nature de ce travail doctoral est particulière. En effet, à ce jour nous gardons toujours le contact avec toutes nos répondantes à travers un groupe sur le réseau social Facebook. Nous échangeons encore lors des occasions sociales, familiales et parfois juste pour prendre des nouvelles. Il ne s’agit pas d’une amitié, mais plutôt d’un véritable lien de confiance qui a trouvé racine dans la sensibilité des sujets abordés lors des entretiens. La confiance perdure après la recherche et se traduit par exemple par une acceptation sans réserve de signer notre demande de lettres de consentement écrites. Photographie 48: le groupe messenger sur Facebook toujours actif En signe de leur volonté de maintenir le lien, nos informantes nous ont offert des cadeaux après la production des données et avant de quitter la dernière réunion de groupe. Mieux encore, nous avons reçu des invitations pour revenir en famille les visiter pendant les vacances. Photographie 49 : Des cadeaux et des invitations de nos répondantes 465 Ces attentions illustrent la force du lien de confiance mais aussi l’efficacité de notre posture bienveillante, non irritante et respectueuse avec le terrain sensible. Jusque là, nous avons mis en lumière ce que le travail réflexif nous a permis de mettre en place pour gérer la subjectivité du chercheur dans l’installation du lien de confiance avec le terrain sensible. A présent, nous expliciterons les effets de notre pratique réflexive sur l’établissement et l’analyse de nos résultats. Ceci rend visible le passage de ce travail doctoral à une recherche care. 1.4. De la sensibilité du terrain à une recherche care La pratique réflexive nous a permis d’interroger l’éthique des résultats de la recherche, c’est-àdire l’évaluation des dangers inhérents à la construction du savoir et à sa diffusion (Blanchet, 2009), et ce à la fois par rapport aux répondantes mais aussi par rapport au chercheur lui-même. Nous avons dès lors mis en perspective une gestion de la vulnérabilité du terrain afin de pouvoir produire de la connaisance, ainsi que le respect d’une certaine charte pour la diffuser. Tout d’abord, il fallait reconnaitre la vulnérabilité duale du terrain, c’est-à-dire la reconnaissance du besoin d’empathie, aussi bien pour les répondantes que pour le chercheur, (Tronto, 2008). Nous l’avons bien compris, au regard de l’hypersensibilité des répondantes, mais aussi de la charge mentale que nous avons dû gérer, et ce dès les premiers entretiens. Pour produire de la connaissance, il a fallu gérer cette double vulnérabilité sur ce terrain sensible en mettant en œuvre des actions en direction d’autrui (les répondantes) et de soi même (le chercheur) (Fischer et Tronto, 1990). Ces actions devaient prendre en considération une vulnérabilité mutuelle, la comprendre, la canaliser pour pouvoir mener à bien la recherche et produire de la connaissance. Ensuite, nous avons dû respecter un certain nombre de règles se rapportant à l’anonymat et à la confidentialité de la recherche, mais aussi à notre responsabilité en tant que chercheur sur un terrain sensible. Plus précisément, notre façon de faire s’est articulée autour de deux grandes actions qui ont permis un certain dosage entre empathie et préservation de l’autre et de soi : l’accueil du sensible et la réception du sensible. 466 L’accueil du sensible d’autrui s’est articulé autour des actions de gestion de la vulnérabilité des répondantes lors de la production de la connaissance et de sa diffusion. Alors que la réception du sensible a suscité l’auto-gestion de la sensibilité de soi. Cette dernière s’est focalisée sur le fait de prendre soin de soi-même en tant chercheur pendant tout le travail de recherche.	43,075
https://openalex.org/W2565699046_39	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	7,666	12,656	344 El TSA ha señalado que no puede haber responsabilidad cuando la publicidad se basa en informaciones contenidas en registros públicos, ya que en esos casos la protección dada a la privacidad es de poco interés. Cox Broadcasting 494, 95 S.Ct. 1029; Jenkins v. Dell Publishing Co., 251 F.2d 447 (3d Circ.) (1958). Más recientemente, el mismo criterio se utilizó en Florida Star v. B. J. F., 491 U.S. 524 (1989). Asimismo, la jurisprudencia americana ha señalado que no hay indicios que permitan afirmar que la primera enmienda protege de diferente manera a los historiadores que a quienes reportean hechos actuales, ni que el ejercicio de esta libertad se ve afectada por el paso del tiempo. Bridges v. California, 314 U.S. 252, 62 S.Ct. 190, 86 L.Ed. 192 (1941). Frente a este aspecto particular ha señalado que ante la ausencia de criterios que permitan determinar cuándo una noticia era lo suficientemente vieja como para no recibir la protección de la primera enmienda, podía derivarse en situaciones de autocensura (derivados de la incertidumbre del informador frente a la legitimidad de sus actuaciones), que fue precisamente lo que quiso evitar la TS en Cox Publishing and The Florida Star (Uranga v. Federal Publication Inc, Idaho, Supreme Court No. 27118. (14 de Febrero de 2003). Por lo mismo, debe confiarse en el criterio de quienes deciden publicar (Cox Broadcasting, 420 U.S. 496, 95 S.Ct. 1029). En el mismo sentido se manifiesta uno de los textos jurídicos más influyentes en la tradición jurídica norteamericana, el Restatement of Torts (Second), que señala que no habrá responsabilidad cuando se difundan hechos que ya son públicos. Restatement of Torts (Second), párrafo 652D cmt. b volumen 3(1977). Cabe señalar que tampoco en Europa existe en la actualidad un concepto legal preciso de privacidad o vida privada, pese a los esfuerzos del TEDH por perfilarlo a través de sus fallos. Observando un desarrollo expansivo del mismo, podemos decir que este derecho protegería la inviolabilidad del domicilio, la protección de una esfera privada frente a las intromisiones de los poderes públicos, las comunicaciones individuales y privadas con terceros, así como la reputación y la propia imagen. Podría alcanzar el derecho de cada individuo de conducir libremente su vida. También podría referirse a otras materias, como la soberanía personal sobre la integridad personal (sería un asunto privado, y permitiría dar protección, por ejemplo, a las prácticas sadomasoquistas consentidas, por ejemplo sentencia de 19 de febrero de 1997, Laskey, Jaggard y Brown, apartado 36. Para algunos también esto incluye el derecho de la mujer a la libre interrupción del embarazo, o el derecho de los transexuales a cambiar quirúrgicamente de sexo, pero no hay claridad sobre estos puntos. José MARTIN Y PÉREZ DE NANCLARES, “Comentario al artículo 7 de la Carta de los Derechos Fundamentales de la Unión Europea”. Dirigido por Araceli MANGAS, Separata Título II, sobre las libertades, Fundación BBVA (2008) 546 Esta postura simplifica el trabajo jurisprudencial y refuerza la libertad de expresión, pero lo hace a costa de extender la protección constitucional a situaciones claramente dañosas y absurdas. A nuestro entender, ello obedece a un juicio de ponderación equivocado345, pues parece de sentido común plantear el “interés público” y su perdurabilidad como un asunto dinámico y circunstanciado346. Bajo esta mirada se abre un espacio justificado a la protección privilegiada de otros derechos, como ha sido observado con más claridad en la tradición europea, que ha reconocido un factor temporal en la protección de la privacidad, vinculado generalmente al paso del tiempo y a la pérdida del interés público de la información que generalmente conlleva347. Por otro lado, existe una vertiente que entiende la privacidad como la posibilidad de controlar el flujo de informaciones sobre uno mismo, especialmente cuando el interés público que justificaba la protección reforzada de la libertad de expresión e información ha disminuido. Para entender lo anterior, conviene detenerse un momento en la evolución del concepto de privacidad. Como es sabido, sus primeros desarrollos fueron realizados dentro de la tradición anglosajona, al alero del derecho de propiedad 348. Luego de numerosas elaboraciones y ajustes doctrinarios y jurisprudenciales, esta concepción ha permitido mantener en la esfera de lo “privado” (como opuesto a público) aquellas informaciones relativas a la propia persona, estableciendo una gradualidad entre ciertas informaciones “íntimas”, que gozan de máxima protección (vida sexual, afectiva, antecedentes médicos, entre otros); aquellas sencillamente “privadas” (actividades profesionales, etc.), y otras que gozan de una mínima protección (ser visto en la calle, la fachada de una casa, las acciones de las autoridades políticas, etc.). La privacidad así entendida se vincula al derecho a ser desconocido, a contar con un espacio de acción donde nadie puede interferir, a ser dejado solo 349. Dada su Baste con recordar el criterio sentado por el TSA en el caso Cox Broadcasting, citado reiteradamente a lo largo de este trabajo, y que justificó la publicidad del nombre de una víctima de violación y que fuera luego reiterado en Florida Star v. B. J. F., 491 U.S. 524 (1989). 346 De hecho, el paso de los años normalmente hará disminuir, total o parcialmente, el interés público asociado a un hecho. La importancia de la pronta divulgación informativa de los asuntos de interés públicos es precisamente lo que justifica la tolerancia a cierto margen de error en las mismas por parte del comunicador. Al respecto, ver STC 6/1988, FJ 5; STC 28/1996, FJ 3; STC 2/2001, FJ 6; STC 158/2003, FJ 6; STC 160/2003, FJ 5; y STC 216/2006, FJ 8. En la jurisprudencia americana, puede consultarse Nueva York Times Co. v. Sullivan, 376 U.S. 254 (1964). En la Corte Europea de Derechos Humanos, la CEDH Nº 28525/95, Unabhängige Initiative Informationsvielfalt v. Austria, de 26 de febrero de 2002, párrafos 39 y 40. En su símil interamericano, la CIDH, Olmedo Bustos y otros v. Chile, sentencia de 5 Febrero 2005; y CIDH, Kimel v. Argentina, sentencia de 2 de Mayo 2008. 347 Patrizia PALERMO (2010), ob. cit., nota 304, p. 278. 348 Dadas las condiciones culturales y jurídicas de dicho pasado, no existía la posibilidad real de construir efectivamente este derecho desde otra base. Por ello, más que de crítica, su desarrollo al alero del derecho de propiedad y del domicilio debe ser motivo de gratitud y considerarse como un antecedente de las versiones más modernas de privacidad. 349 Esta expresión fue popularizada por el célebre artículo sobre “Privacidad” de Warren y Brandeis, quizá el más influyente a nivel mundial sobre el tema. Samuel D. WARREN y Louis D. BRANDEIS, “The Right to Privacy”, Harvard Law Review, Vol. IV, No. 5 (1890). En el mismo sentido se pronuncia MILLER, quien literalmente dice que “el derecho a la privacidad se 345 547 íntima relación con el concepto patrimonial de propiedad, y más particularmente a la idea del “domicilio”, usualmente se le ha protegido por la vía del derecho civil (“torts”)350. Sin embargo, la privacidad ha visto también un desarrollo diferente y más amplio, observable especialmente en el continente europeo, que se vincula con el derecho a excluir del dominio público ciertas informaciones, con el objeto de proteger y facilitar el derecho de toda persona a vivir como dueña y arquitecta de su propio destino351. Bajo esta concepción, construida antes sobre la idea de dignidad humana que de propiedad, puede justificarse el control del uso público de aquellas informaciones que puedan afectar al individuo en sus relaciones con los demás, sobre todo cuando ellas se vinculan con errores o hechos del pasado. Por ello, proteger la presenta como el derecho de estar solo y ahora debe considerarse desde la perspectiva de la “habilidad individual para controlar la circulación de información relativa a él mismo (…)”. Arthur R. MILLER, “The Assault on Privacy: Computers, Data Banks and Dossiers”. University of Michigan Press (1971). Por su parte, el TCE ha señalado en su STC 134/1999, FJ 5 que “El art. 18.1 no garantiza una “intimidad” determinada, sino el derecho a poseerla, a tener vida privada, disponiendo de un poder de control sobre la publicidad de la información relativa a la persona y su familia, con independencia del contenido de aquello que se desea mantener al abrigo del conocimiento público. Lo que el art. 18.1 garantiza es un derecho al secreto, a ser desconocido, a que los demás no sepan qué somos o lo que hacemos, vedando que terceros sean particulares o poderes públicos, decidan cuáles sean los lindes de nuestra vida privada, pudiendo cada persona reservarse un espacio resguardado de la curiosidad ajena, sea cual sea lo contenido en ese espacio. Del precepto constitucional se deduce que el derecho a la intimidad garantiza al individuo un poder jurídico sobre la información relativa a su persona o a la de su familia, pudiendo imponer a terceros su voluntad de no dar a conocer dicha información o prohibiendo su difusión no consentida”. En el mismo sentido se pronuncian las STC 144/1999, STC 115/2000, STC 185/2000 y STC 127/2003, entre otras. 350 Nuevamente recomendamos seguir el desarrollo de este tema realizado en “La vida privada de las figuras públicas. El interés público como argumento que legitima la intromisión en la vida privada”. Ignacio COVARRUBIAS CUEVAS (2013), ob. cit., nota 321. 351 ROLLA señala que esta profunda evolución “supera, en primer lugar, una concepción de la privacy estrechamente vinculada al derecho de propiedad (entendido como ius excludendi alios) y orientada de modo predominante a la tutela de la inviolabilidad del propio domicilio (…) hasta adquirir un más amplio alcance: trasladando, primero, la atención del “tener” al “ser” (…) asumiendo, luego, en plena sociedad global de la comunicación, un significado más amplio como derecho al control de las informaciones relacionadas con uno mismo”. A su vez ello implica el paso desde una concepción negativa de la privacidad a una que requiere numerosas actuaciones positivas. Bajo esta lógica, la intimidad “encierra también el aspecto "positivo" de poder desarrollar libremente la propia personalidad. La garantía de la vida privada tiende, en esta acepción amplia, a identificarse con el derecho a la tutela de la propia identidad (sexual, de conducta, ideal, familiar, etcétera). El respeto de la privacy se transforma, pues, en la necesidad de tutelar las opciones de vida frente a toda forma de control público o de "estigmatización social" (citando a Lawrence M. FRIEDMANN, “The Republic of Choice: Law, Authority, and Culture”, Cambridge University Press (1990). Continúa señalando que “la tutela de la vida privada cumple con ello un papel instrumental para la libre construcción y establecimiento de relaciones y vínculos sociales (…). En otros términos, el derecho a la vida privada responde a la exigencia de que toda persona pueda construir autónomamente su propia personalidad, sin que desde fuera se le impongan los roles o funciones que deba desarrollar”. Giancarlo ROLLA (2002), ob. cit., nota 305, pp. 150-151. En un sentido similar, Stefano RODOTÀ, “La Pacifica Revoluzzione della Privacy”, Questione Giustizia, No. 2 (2005), p. 228. 548 autonomía y la libertad individual supone la protección del sujeto con sus características distintivas actuales, en correspondencia con su imagen pública352. De esta forma, esta concepción de la privacidad fundamenta el derecho al olvido, favoreciendo a la vez la tutela del pluralismo y de la autonomía personal frente a las limitaciones que puedan provenir del exterior. El reciente fallo del Tribunal de Justicia de la Unión Europea sobre el caso Google353, dará un importante refuerzo y fundamento de autoridad a esta postura. En los últimos años han existido muchas señales jurisprudenciales en este sentido, aunque por las razones ya señaladas, ellas han sido mayores en el espacio europeo que en la tradición anglosajona. Francia quizá sea el país donde la protección de la privacidad frente a la libertad de expresión alcanza su mayor protección, y ha aceptado la consideración al elemento temporal como relevante en la protección de la privacidad en numerosas oportunidades354. Así sucedió, por ejemplo, frente al reclamo presentado por una ex amante del famoso asesino en serie Henri Landru, frente a un film que la mencionaba, sosteniendo que ello correspondía a un periodo muy antiguo y dramático de su vida privada y que hubiera querido olvidar355. El mismo criterio se utilizó frente a la publicación de la autobiografía de un famoso delincuente (Mesrine), en la que una antigua pareja era mencionada, afectando, según ella, la reinserción social que había logrado con el transcurso del tiempo 356, o ante el caso del cantante Jean Ferrat, quien recibió protección judicial luego de alegar por la revelación, por medio de la prensa, de su nombre y domicilio, luego de que ya se encontraba retirado357. Un caso aún más interesante se originó por la inclusión, en un juego de mesa tipo TRIVIA, de una pregunta sobre el nombre del médico que en compañía de una enfermera había sido sorprendido robando un banco. El médico intentó la prohibición de venta y el retiro del juego. El Tribunal de París acogió la demanda rechazando el argumento de que los hechos ya publicados o los asuntos judiciales fueran por si mismos noticiables en forma indefinida, señalando, además, que el tiempo transcurrido había quitado el interés público a los hechos, por lo que ya no existía una necesidad social de contar con información sobre ellos. De esta manera, se consideró que la posterior vida del médico y su protección debían imponerse sobre la liberta de prensa358. 352 Massimiliano MEZZANOTTE (2009), ob. cit., nota 280, pp. 69-70, 79 y 81. 353 La cuestión preliminar planteada por la Audiencia Nacional Española, pero que se pronuncia sobre los aspectos más relevantes de la normativa aplicable, fue resuelto por la Sentencia del Tribunal Europeo de Justicia (TJUE) de 13 de mayo de 2014, dictado por la gran sala y recaída (asunto C-131/12, planteado por la Audiencia Nacional, mediante auto de 27 de febrero de 2012, en el procedimiento entre Google Spain, S.L., Google Inc. y Agencia Española de Protección de Datos (AEPD), Mario Costeja González. 354 ROSEN afirma que las raíces intelectuales del derecho al olvido provienen de la legislación francesa, que reconoce un “droit à l´oubli”, relativo a la rehabilitación de delincuentes y su resocialización. Jeffrey ROSEN, “The Right to Be Fortgotten” (2012). 355 Tribunal de Gran Instancia de Sena, 4 de octubre de 1965. 356 Tribunal de Gran Instancia de Paris, 6 de diciembre de 1979. 357 Corte de Apelaciones de Paris, 15 de mayo de 1970. 358 Tribunal de Gran Instancia de Paris, Sala Primera, 25 de marzo de 1987. 549 En 2004, siguiendo el mismo criterio, el Tribunal de Casación revocó una sentencia de la Corte de Apelaciones que sostenía que la publicación, en una película, de hechos relativos a un accidente familiar afectaba la vida y la personalidad de uno de sus miembros359. En otro caso, la Corte de Casación, pese a que rechazó dar protección solicitada por una mujer mencionada en un libro como compañera de un colaboracionista, señaló como criterios para legitimar la publicación el que las publicaciones originales (en caso que las hubiera) hubiesen sido realizadas al amparo de la ley, y que persistiera un interés actual que justificara publicación360. En Italia, el derecho al olvido era ya mencionado en una sentencia del tribunal de Casación de 1958361. 40 años más tarde, el mismo tribunal ya lo reconocía en forma expresa, refiriéndose al “justo interés de cada individuo de no estar indefinidamente expuesto a datos que afectan negativamente su honor o reputación, relativa a la reiterada publicación de una noticia divulgada en el pasado”362. Este criterio ha sido aceptado muchas veces frente a las solicitudes de actrices para que no se publiquen fotos de desnudos realizadas en su juventud, porque se "referían a una fase de su carrera ya superada en su nueva situación artística o profesional"363. Con estos ejemplos, de los que pueden encontrarse casos similares en otros países europeos364, abordamos una forma de enfrentar este problema, típicamente europea. Pero en EE.UU., por el contrario, la amplia protección dada a la difusión libre de información y la concepción de la privacidad en su forma más restrictiva y vinculada al derecho de propiedad, han hecho menos permeable su ordenamiento 359 Corte de Casación, Sala Civil 2, de 3 de Junio de 2004. 360 Sentencia de la Corte de Casación Civil del 20 de Noviembre de 1990. Pueden encontrarse referencia a estos casos y a otros en Massimiliano MEZZANOTTE (2009), ob. cit., nota 280, p. 216 y ss. 361 Sentencia de la Corte de Casación del 13 Mayo de 1958, Nº 1563, conocida como el caso Caruso. Según MEZZANOTTE, la jurisprudencia la reconoció por primera vez en un obiter dicta, en 1973, donde hacía referencia a los artículos 8 y 10 de la CEDH, y del mismo modo, se veía como a veces un límite absoluto frente a la libertad de expresión, o bien como un aspecto a balancear en cada caso concreto. Ibid., pp. 82 y 98 (cita 67). 362 Sentencia de la Corte de Casación de 9 Abril de 1998, Nº 3679. Más recientemente, vemos sentencias que manifiestan “una voluntad orientada al reconocimiento de un derecho, cuyos contornos son todavía poco claros”. Ibid., p. 277 (haciendo expresa referencia a la sentencia Nº 45051 de 24 de noviembre del 2009, dictada por la sección quinta de la Corte de Casación Penal Italiana). 363 Pretura de Roma de 10 de febrero de 1988. Citado por Giancarlo ROLLA (2002), ob. cit., nota 305, p. 165 (nota al pie No 4). 364 En España, por ejemplo, la STC 52/2002, estimó que la publicidad mediática dada a los antecedentes penales de una persona por una violación acaecida hace doce años, y por un arresto menor, eran superfluas e innecesarias, dado que la noticia fundamental se había centrado en descartar su participación en el doble crimen (homicidio). Añadir aquellas referencias, que afectan a su probidad, reputación y dignidad personal, era innecesario, al “no poder calificarse de veraz y carecer de relevancia e interés público”. (FJ 7 y FJ 8). En Alemania, por ejemplo, el llamado caso Lebach (BVerfGE 35, 2002, 1973). 550 a una restricción fundada en el factor temporal o “derecho al olvido” 365. El tratamiento dado a este tema por la doctrina y jurisprudencia de este país nos parece especialmente relevante, dada la enorme influencia del derecho americano en el mundo, y en particular, en su desarrollo sobre la libertad de expresión, por lo que nos detendremos brevemente sobre algunos aspectos del mismo. El criterio más clásico y dominante en el sistema americano parece ser el adoptado por el Tribunal Supremo de los Estados Unidos en el caso Briscoe v. Reader´s Digest Association, Inc., donde la publicidad dada al autor de un delito cometido 11 años atrás a propósito de un nuevo ilícito, se consideró legítima, dada la prevalencia del interés de la noticia sobre los demás derechos afectados366. Un criterio similar se utilizó en el caso Jenkins v. Dell Publishing Co., donde la familia de una víctima de asesinato autorizó y facilitó la cobertura periodística del mismo, pero luego protestó ante la inclusión del material en un libro de crímenes. El Tribunal Supremo de los Estados Unidos señaló que una vez que un asunto era noticia, no resulta factible ni deseable para un tribunal comenzara a distinguir entre noticias que buscaban informar y aquellas que buscaban entretener367. Un caso levemente diferente afectó a William J. Sidis, conocido desde muy corta edad por sus asombrosas facultades intelectuales, y que con el correr del tiempo, desarrolló una vida más bien discreta. En 1937 el “New Yorker” publicó un artículo titulado “¿Dónde está ahora?”, dando origen a un reclamo por invasión a la privacidad. El amplio alcance dado a la libertad de expresión y al concepto de interés público en Estados Unidos llevó a que el Tribunal de Apelaciones de Nueva York señalara que "lamentablemente o no, las desgracias y debilidades de los vecinos y" figuras públicas "son temas de gran interés y debate con el resto de la población"368. Con ello, el criterio de “figuras públicas” fue desplazándose desde el ámbito de las acciones de gobierno a la vida deportiva, al mundo del espectáculo, de la ciencia, los negocios, etc.369 El criterio de máxima publicidad parece haberse consolidado en el caso “Cox Broadcasting370”, que declaró inconstitucional una ley de Georgia, que prohibía la publicación del nombre de las víctimas de violación. Este es uno de los casos más citados y de mayor influencia doctrinaria y jurisprudencial en lo referido a la primera enmienda. Sin embargo, este criterio no parece monolítico, presentándose algunas soluciones minoritarias en sentido contrario. Uno de ellos, por ejemplo, favoreció a una dueña de casa de un pueblo del interior de los Estados Unidos, que luego de ser involucrada en un caso de 365 WERRO señala que en EE.UU. “el desarrollo de un derecho a ser olvidado (…) no viene, como en Europa, derivado de un equilibrio entre los derechos reconocidos en la Constitución, sino más bien, de una serie de intentos por parte de los diversos estados de forjar para sus ciudadanos una esfera de la intimidad inviolable respecto de los medios de comunicación (…)”. Franz WERRO (2009), ob. cit., nota 306, p. 306 366 Briscoe v. Reader's Digest Association, Inc., 4 Cal.3d 529. 367 Jenkins v. Dell Publishing Co., 251 F.2d 447 (3d Circ.) (1958). 368 Sidis v. F-R Publishing Corp., 113 F.2d 806 (2d Cir.). 369 Ignacio COVARRUBIAS CUEVAS, “Notas Críticas a la Figura del Personaje Público como Criterio Legitimador para la Intromisión en la Vida Privada de las Personas”, Revista del Centro de Estudios Constitucionales, Vol. 3, No. 2 (2005), p. 169 370 Cox Broadcasting Corp. v. Cohn, 420 U.S. 469 (1975). Florida Star v. B. J. F., 491 U.S. 524 (1989). 551 homicidio -del que finalmente salió libre de cargos- fue sindicada por la prensa como principal sospechosa, al descubrir que durante su juventud había sido trabajadora sexual. Como cabía imaginar, dicha circunstancia no era conocida por parte de su pequeña comunidad, generándole perjuicios de todo tipo, entre los que cabe imaginar un aumento de las sospechas de culpabilidad por parte de la opinión pública, y probablemente del jurado. Pocos años después, su caso fue argumento de una película, llamada “The Red Kimono”, que incluso utilizó su nombre de soltera. Frente a las reclamaciones de la afectada, el TS-California consideró que si bien los datos eran públicos y reales, cualquier persona que hubiera rehecho su vida tenía derecho a la búsqueda de la felicidad y a no sufrir ataques innecesarios, y que el uso de su nombre verdadero no aportaba información para ilustrar el caso371. Más recientemente, el caso US Dept. of Justice v. Reporters Committe372 se refirió a la “Freedom of Information Act” (FOIA), que permite obtener acceso a las informaciones de las instituciones públicas. El caso se originó por la negativa dada por el Departamento de Justicia de entregar a un grupo de periodistas informaciones recopiladas por el FBI que vinculaban a cuatro hermanos acusados de obtener contratos con el Estado en forma ilegítima, con la asistencia de un congresista. El FBI sólo entrego un “rap sheet” relativo a un hermano ya fallecido, y dado que durante el transcurso del juicio fallecieron otros dos, se liberó la información que se refería a ellos. Sin embargo, se aclaró expresamente que respecto del sobreviviente sólo se entregaría información si hubiera un interés público relativo a delitos financieros (cosa que no ocurría) y no sobre otras materias. El Tribunal Supremo de los Estados Unidos validó el criterio del FBI, señalando la privacidad estaba protegida por la excepción 7 c) de la “Freedom of Information Act” (FOIA) (vinculada al factor temporal), agregando que el principal propósito de esta normativa era permitir el público escrutinio público de los actos de gobierno, y no el obtener información privada de otros ciudadanos. Por ello, aunque el registro no era “privado”, podían justificarse la protección de intereses individuales que buscaran limitar su publicidad373. Un caso aún más interesante se originó en la sentencia que cuestionó la legitimidad de una publicación que hacía referencia a actividades homosexuales entre adultos y adolescentes acontecidos a mitad de los años 50, generando gran revuelo (una de las personas involucradas fue luego expulsada de la Academia de West Point). La sentencia consideró que el caso era muy distinto al precedente de Cox Broadcasting374 y de toda la jurisprudencia nacida a su amparo, señalando que en algunos casos el interés público era dudoso375. Dado el giro que esta sentencia podía implicar frente a la tradicional comprensión de la primera 371 La reclamante no utilizó en esta instancia el argumento del derecho a la privacidad ni a la propia imagen, pues ellos no habían sido acogidos previamente. 372 US Dept. of Justice v. Reporters Committee, 489 U.S. 749 (1989). 373 Sin embargo, ello no significa que estas directrices sean efectivamente seguidas, pues el gobierno a menudo guarda los archivos en bibliotecas, donde son fácilmente accesibles. Massimiliano MEZZANOTTE (2009), ob. cit., nota 312, pp. 241- 242. 374 Cox Broadcasting Corp. v. Cohn, 420 U.S. 469 (1975). 375 Uranga v. Federal Publication Inc, Idaho, Supreme Court (21 de junio de 2001). 552 enmienda, fue objeto de un recurso especial ante el TSA-Idaho (“rehearing”), la que validó tanto la publicación como los criterios de las instancias inferiores, reiterando las dudas sobre cómo el paso del tiempo podría afectar el interés público de un hecho, así como la dificultad de determinarlo 376. Pese al criterio final esgrimido en este caso, todo indica que la problemática ya se encuentra planteada y ha dejado de ser un asunto indiscutible. 6.- La protección de datos personales como una forma de derecho al olvido En las últimas décadas, la sofisticación de los medios técnicos de acumulación, conservación y acceso a la información ha levantado la problemática del “olvido” a materias donde antes no se habían planteado mayores situaciones de conflicto. Ello se ha visto profundizado por los mecanismos con que cuentan algunos sistemas de internet para registrar antecedentes personales de los usuarios, como por ejemplo las páginas visitadas, las compras, las direcciones IP, las claves y números de tarjetas de crédito, entre otras. Las nuevas tecnologías han permitido la creación de bases de datos y sistemas de información de tal magnitud y potencialidades, que su sola existencia podría reducir significativamente los espacios de privacidad377. Todo lo anterior produce problemáticas sociales nuevas que inquietan a buena parte de la población378. Pese a las limitaciones técnicas, jurídicas y fiscalizadoras que plantea la protección de los derechos afectados por esta vía, creemos necesario realizar un esfuerzo por enfrentarlos y lograr, poco a poco, mayores niveles de protección379. 56 Uranga v. Federal Publication Inc, Idaho, Supreme Court No. 27118 (14 de febrero de 2003). Respecto a la falta de necesidad de que su nombre hubiese sido revelado, la Corte respondió citando numerosa jurisprudencia, que cada elemento de la noticia era relevante. 377 El sólo aumento cualitativo de las facilidades a la hora de compilar información, podría alterar las reglas tradicionales sobre esta materia. Así lo ha señalado incluso el TSA en Departement of Justice v. Reporters Committee for Freedom of the Press, 489 US 764-769 (1989). En un sentido similar, Westbrook v. County of Los Angeles (1994) 27 Cal. App. 4th 157 [32 Cal. Rptr. 2d 382]. 378 Según la Comisión Europea, 9 de cada 10 europeos están preocupados de cómo se utilice la información obtenida a través de aplicaciones de teléfonos celulares, y 7 de cada 10 lo están acerca de cómo dicha información puede ser posteriormente usada. European Commission, Press Release of 28, January 2014. Según una encuesta realizada por EuroBarómetro en 2011, el 74% de los europeos consideran la publicidad de sus informaciones personales como un rasgo inherente de la vida moderna, pero al mismo tiempo, un 43% cree que han sido requeridos de mayor información de la que era necesaria al usar la red. Cerca de un 60% de los usuarios lee las condiciones de privacidad, y de ellos, un 70% ajusta su conducta a ella. Sólo la mitad de los usuarios conocerían las condiciones de recopilación de sus antecedentes para usos posteriores cuando usan internet. Sólo un 18% siente control total sobre la información disponible, y un 70% se muestra preocupado por el uso que las compañías puedan hacer de la información requerida para otros propósitos. Un 75 % de los europeos se manifestó partidario de eliminar la información personal entregada en internet cuando ellos lo decidieran. Información extraída de: Special EuroBarometer 359, Attitudes on Data Protection and Electronic Identity in the European Union (June 2011), disponible en http://ec.europa.eu/public_opinion/archives/ebs/ebs_359_en.pdf (fecha de consulta: 9 de mayo de 2014). 379 Ya en el año 1968, la Conferencia Internacional de Derechos Humanos (ONU) realizada en Teherán manifestó su preocupación sobre los riesgos de violación de los derechos, que implicaba el avance científico y tecnológico. El Convenio 108 del Consejo de Europa establece 553 Con este propósito, la Comunidad Europea aprobó la Directiva Nº 46/95, sobre “Tratamiento y libre circulación de datos personales”380, que entre otras cosas, llamó a la modificación de las normas de los países miembros con el objeto de dar un espacio mayor al derecho a la privacidad, reconociendo incluso la debida consideración que debía hacerse al elemento histórico al momento de buscar un adecuado equilibrio con otros derechos. Dicha directiva también buscó que los Estados miembros garantizaran, con arreglo a las disposiciones de la Directiva, “la protección de las libertades y los derechos fundamentales de las personas físicas, y en particular, el derecho a la intimidad, en lo que respecta al tratamiento de los datos personales”381. En el mismo sentido se han pronunciado la Directiva 2002/58/CE, de 12 de julio de 2002, modificada el 23 de abril de 2009 por la Directiva 2009/22/CE 382, las cuales se encuentran en pleno vigor y obligan a los Estados miembros a adoptar las disposiciones necesarias para darles cumplimiento. Adicionalmente, y en base a lo ya señalado por la Directiva 46/95/CE, la Comisión Europea, ha elaborado una Propuesta de Reglamento Europeo de Protección de Datos de personas físicas, donde establece nuevas protecciones y garantías, y que se espera reemplace a la primera una vez que sea aprobado. Entre ellos, cabe mencionar la creación de nuevos derechos para los ciudadanos, entre los que se incluye el derecho a “la supresión de datos bien porque ya no son necesarios conforme a la finalidad para la que fueron recabados ya sea porque el interesado ha revocado su consentimiento para el tratamiento; porque ha expirado el plazo para el tratamiento legal de los datos; porque el interesado ha ejercitado su derecho de oposición, o bien porque el tratamiento de los datos no se está realizando conforme al mismo Reglamento383. desde 1981 el derecho a la protección en lo que respecta al tratamiento automatizado de los datos personales. El concepto de datos personales no está vinculado a la idea de intimidad propia de la privacidad. Es más amplio y abarca cualquier tipo de información de tipo personal, y supone una suerte de “autodeterminación informativa”. Así lo planteó primeramente el documento de trabajo titulado “Programa de Estocolmo. Una Europa Abierta y Segura que Sirva y Proteja al Ciudadano”, información que se encuentra disponible en http://eurlex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:C:2010:115:0001:0038:es:PDF (fecha de consulta: 10 de mayo de 2014). Luego, el 4 de noviembre de 2010, la Comisión fijó una estrategia para reforzar las normas de protección de datos de la UE (IP/10/1462 y MEMO/10/542) y solicitó reacciones a sus ideas y llevó a cabo una consulta pública por separado para revisar la “Directiva sobre Protección de Datos” (95/46/CE). 380 Como antecedente de la misma, los artículos 7 y 8 de la Carta de los Derechos Fundamentales de la Unión Europea reconocen el respeto de la vida privada y la protección de los datos de carácter personal, y el Convenio N° 108 del Consejo de Europa, de 28 de enero de 1981, fue la primera norma internacional establecida sobre el tratamiento automatizado de datos de carácter personal. 381 El artículo 2.a de la Directiva 95/46, define dato personal como “toda información relativa a una persona física identificada o identificable”. De esta manera, el objeto protegido por este tipo de “derecho al olvido” puede ser mayor que el que nace de la ponderación entre derechos fundamentales. Se han incorporado a las propuestas elementos moduladores en este sentido (requisitos, excepciones, conceptos abiertos que quedan entregados al criterio judicial, etc.). 382 Existen además disposiciones específicas para la protección de los datos personales en el marco de la cooperación policial y judicial en materia penal (Decisión Marco 2008/977/JAI). 383 Artículo 17 del proyecto de Reglamento de Protección de Datos Personales de la CE. 554 El 21 de octubre de 2013, el Comité del Parlamento Europeo de Libertades Civiles, Justicia y Asuntos del Parlamento Europeo aprobó la propuesta de Reglamento de Protección de Datos. El 12 de Marzo de 2014, el Parlamento aprobó la propuesta, incorporándole algunas modificaciones. En lo que respecta a nuestro trabajo, la expresión que hacía expresa referencia al derecho al olvido fue eliminada, pero fue mantenida en forma indirecta por la vía del “derecho al borrado de datos”. Entre otras medidas el Reglamento exige consentimiento explícito y permanente para la publicación de datos (en vez de presumirla); propicia la utilización de pseudónimos cuando el conocimiento de la identidad no sea necesario o se trate de información personal sensible; limita la transferencia de datos a espacios que estén fuera de la jurisdicción europea; establece acciones de protección que deben ser resueltas en plazos muy breves (24 horas desde la solicitud); además de contemplar importantes sanciones (que pueden llegar hasta 100 millones de euros o el 5% del volumen de negocio anual de los infractores). Algunas de las medidas planteadas en la propuesta de Reglamento parecen vincularse con el debate que ha generado un caso cuyo reciente fallo afectó a un gigante del mundo digital (Google) respecto a la posibilidad que ofrece de acceder, por su intermedio, a una serie de datos del pasado, publicados muchas veces en forma legítima. Este fallo había sido muy esperado por los estudiosos de las regulaciones de internet y la privacidad384. El caso en cuestión fue fallado mientras terminábamos la redacción de este artículo, y se pronuncia sobre los interrogantes más delicadas de la posible regulación sobre protección de datos personales: la responsabilidad y colaboración que cabe a los motores de búsqueda, que en la práctica, son los que hacen posible el acceso facilitado a todo tipo de informaciones y base de datos, y adicionalmente, la problemática que plantea cualquier intento de reglamentar un sistema de carácter transcontinental. El juicio tiene su origen en el reclamo presentado por el ciudadano español Mario Costeja González, quien luego de ser ejecutado por una deuda en 1998, volvió a ver su nombre en la web 10 años después, luego de que el periódico La Vanguardia (donde había sido publicado el aviso de remate), diera acceso online a toda su hemeroteca, generando su indexación automática por parte de los motores de búsqueda de internet y el acceso masivo y facilitado del público a esta información. 384 Sentencia del Tribunal Europeo de Justicia (TJUE) de 13 de mayo de 2014, dictado por la Gran Sala, y recaída en el Asunto C‑ 131/12, planteado por la Audiencia Nacional, mediante auto de 27 de febrero de 2012, en el procedimiento entre Google Spain, S.L., Google Inc. y Agencia Española de Protección de Datos (AEPD), Mario Costeja González. SAMPERE se refiere así a este documento: “Por fin llegó el que probablemente haya sido uno de los días (el otro será cuando se pronuncie definitivamente el Tribunal de Justicia de la Unión Europea) más esperado en el “mundillo del dato”: las Conclusiones del Abogado General sobre el asunto de Google Spain SL y Google INC contra la AEPD y un particular. Hace meses, y bajo el título “Google, Abanlex y AEPD en el TJUE: otro punto de vista”, ya ponía de manifiesto que más allá de dar la razón a uno u otros, desde mi humilde opinión, se estaba jugando buena parte de la propuesta de Reglamento de la Unión Europea, probablemente uno de sus artículos más discutidos, que no es otro que el del ámbito de aplicación”. Sobre la influencia de este caso en las propuestas normativas ver Francisco Javier SAMPERE, “Caso “Google” sobre el derecho al olvido: ¿Cómo afecta a la propuesta de Reglamento de Protección de Datos?” (2013). 555 El afectado recurrió primeramente al periódico385, el que señaló que ellos sólo habían dado cumplimiento a un mandato legal, por lo debía dirigir su reclamo al buscador. Google Spain derivó el asunto a Google Inc., en Estados Unidos. Ante la falta de respuesta, remitió el caso a la Agencia Española de Protección de Datos, la que demandó contra Google Spain y Google Inc. para que dejasen de indexar el contenido señalado, aun sabiendo que el motor efectivo de búsqueda estaba en Estados Unidos (California)386. La Audiencia Nacional elevó el caso a instancias europeas387. El pasado 25 de junio de 2013, el Abogado General del Tribunal Europeo respondió preliminarmente a las preguntas planteadas por el tribunal español 388, señalando que Google está sujeta a la legislación de la UE sobre privacidad y protección de datos (sus filiales están establecidas en la UE y ahí realizan su actividad comercial, aunque el tratamiento de datos se realice en otro lugar), pero no está obligado a borrar la información sensible de su índice de búsqueda ni se le puede considerar responsable del tratamiento que se haga de la información, ya que no tiene control sobre los datos personales incluidos en las páginas web de terceros. Por tanto, no existe un derecho a solicitar a los buscadores la no indexación o la desindexación de dicha información389. Finalmente, el 13 de mayo recién pasado, la Gran Sala del Tribunal dictó sentencia declarando que de acuerdo a la Directiva 95/46/CE del Parlamento Europeo y del Consejo “la actividad de un motor de búsqueda, que consiste en hallar información publicada o puesta en Internet por terceros, indexarla de manera automática, almacenarla temporalmente y, por último, ponerla a disposición de los internautas según un orden de preferencia determinado, debe calificarse de “tratamiento de datos personales” (…) y el gestor de un motor de búsqueda debe considerarse “responsable” de dicho tratamiento (Nº 1, parte resolutiva). Asimismo, el Nº 2 (parte resolutiva) señala que las actividades realizadas en un país por la filial implican a la empresa, aunque la sede principal o parte de la actividad se realice fuera de la jurisdicción del país al cual el motor ofrece sus servicios. 385 Procedimiento ordinario 725/2010, seguido ante la Sala de lo Contencioso Administrativo (Secc. 1) de la Audiencia Nacional. 386 La AEPD ha utilizado la estrategia de dirigirse directamente a los buscadores para evitar la indexación de datos en internet, utilizando un criterio sin duda más pragmático, aunque insuficiente, dado que los datos perduran y pueden ser obtenidos y utilizados de otra manera. Otro problema de esta estrategia es que algunos servidores como “Yahoo” o “Bing” niegan la autoridad de esta agencia, por razones de jurisdicción (la primera tiene su sede en EE.UU., la segunda en Luxemburgo). Un análisis exhaustivo de las resoluciones de la Agencia Española de Protección de Datos puede encontrarse en Ramón M. ORZA LINARES y Susana RUÍZ TARRÍAS, “El Derecho al Olvido en Internet”, Neutralidad de la Red y Otros Retos para el Futuro de Internet, Actas del VII Congreso Internacional de Internet, Derecho y Política, Universidad Oberta de Cataluña, Huygens Editorial (2011), pp. 371-389; 387 El Auto de fecha 27 de febrero de 2012 en el que la Sala de lo Contencioso Administrativo (Secc. 1) de la Audiencia Nacional acuerda presentar ante el Tribunal de Justicia de la Unión Europea una cuestión prejudicial que permita aclarar “si la actividad de GOOGLE, como buscador de contenidos de terceras personas, puede considerarse un tratamiento de datos” y, por lo tanto, si la Agencia Española de Protección de Datos Personales (AEPD) puede requerir directamente al buscador, sin dirigirse previa o simultáneamente al webmaster para exigir la retirada de la información (entre otras cuestiones). 388 Dictamen Preliminar del Abogado General del TJUE. 389 Dictamen Preliminar del Abogado General del TJUE, párrafo 138, conclusión Nº 3. 556 Por tanto, señala que “el gestor de un motor de búsqueda está obligado a eliminar de la lista de resultados obtenida tras una búsqueda efectuada a partir del nombre de una persona vínculos a páginas web, publicadas por terceros y que contienen información relativa a esta persona, también en el supuesto de que este nombre o esta información no se borren previa o simultáneamente de estas páginas web, y, en su caso, aunque la publicación en dichas páginas sea en sí misma lícita” (Nº 3 parte resolutiva). Concluye señalando que “si el interesado tiene derecho a que la información en cuestión relativa a su persona ya no esté (…) estos derechos prevalecen, en principio, no sólo sobre el interés económico del gestor del motor de búsqueda, sino también sobre el interés de dicho público en acceder a la mencionada información en una búsqueda que verse sobre el nombre de esa persona390. En los momentos que escribimos estas líneas aún no es posible anticipar las consecuencias que esta sentencia tendrá, pero de seguro marcará un hito importante en el tratamiento de estas materias a nivel mundial. Pero podemos adelantar algunos criterios. El principal de ellos es que pese a que los criterios planteados por el Abogado General del Tribunal Europeo parecían muy razonables, el Tribunal de Justicia de la Unión Europea optó en definitiva por una solución más funcional (más propio de la lógica del balanceo que de la responsabilidad civil clásica fundada en la culpa), privilegiando antes el control de daños que la sanción del acto realmente lesivo y de su autor. En otras palabras: dirigirse a las poderosas empresas que hay detrás de los motores de búsqueda es el mecanismo más fácil para garantizar a los ciudadanos que sus derechos serán respetados, permitiendo dejar los datos inaccesibles en breve tiempo. La sentencia impone también a los buscadores el peso de absorber los miles de reclamos que desde ahora comenzarán a recibir, lo que muy probablemente los llevará a generar las tecnologías adecuadas a ello. Asimismo, el fallo consolida el principio de que cierta información que fue pública en algún momento, puede volver a estar protegida por el ámbito de la privacidad por la disminución del interés público que genera sobre dicho asunto el sólo paso del tiempo. La diferencia de tratamiento dada por la tradición jurídica europea y americana a este tema, claramente visible en las argumentaciones que rodearon a este caso, ha llevado a autores a sostener, incluso antes del reconocimiento de este derecho por parte del Tribunal de Justicia de la Unión Europea, que el derecho al olvido “representa la mayor amenaza a la libertad de expresión en Internet en la década que viene”391. 390 Parte resolutiva, Nº 4. La misma señala que dichos requisitos no se cumplirían, por ejemplo, cuando una persona participa en la vida pública y “la injerencia en sus derechos fundamentales está justificada por el interés preponderante de dicho público en tener, a raíz de esta inclusión, acceso a la información de que se trate”. 391 Jeffrey ROSEN (2012), ob. cit, nota 354, p. 1. 557 7.- Situaciones resistentes al olvido Finalmente, nos referiremos a las situaciones que pese a concurrir el requisito del paso del tiempo y afectar derechos protegidos, ofrecen resistencia al “derecho al olvido”. Aceptando la premisa de que ningún derecho es ilimitado y siempre puede ceder ante otras necesidades e intereses, podemos utilizar como principio general que la especial protección que ofrece la libertad de expresión e información va asociada primeramente a la trascendencia pública de los hechos divulgados, la que dependerá de la relevancia de la materia, su interés público y su capacidad para contribuir a la formación de una opinión pública libre392. Ante la falta de necesidad social de transmitir cierta información, los hechos vuelvan a quedar bajo control de las personas y por lo mismo, protegidas por su derecho a la privacidad393. Sin embargo, este principio puede presentar algunas matizaciones. i.- Disminución y aumento del interés público de un hecho Sin embargo, puede suceder que algunas circunstancias vuelvan actual la información pasada, como cuando la primera publicación y la posterior hubieren sucedido hechos que fueran capaces de volverles su carácter de actualidad (acá comprendemos al interés público informativo como un elemento dinámico). Así pudo observarse, por ejemplo, en el caso Time, Inc. v. Hill, en que la historia de una familia que fue tomada rehén por un grupo de delincuentes prófugos, fue llevada a una novela, y posteriormente, a un exitoso musical en Broadway, haciendo renacer, en buena medida, el interés del público sobre el caso394. Este tipo de situaciones nos muestran que el sólo paso del tiempo no permitirá inclinar la balanza a favor del olvido, volviendo necesario el ejercicio de balanceo. 392 STC 49/2001, FJ 6; y STC 121/2002, FJ 3. 393 La sentencia Nº 3679, del 9 de abril de 1998, señaló que el derecho al olvido era menor si el hecho mismo ya había sido objeto de un gran interés público en el momento en que sucedió (pasado), y por lo mismo o por otra razón, pudiese subsistir un interés público en la noticia, siendo ilegítimos cualquier actividad privada o incluso pública sin relación con los hechos judiciales, o que sólo se vincularan con la vida privada de la persona involucrada en los hechos investigados y conocidos. Massimiliano MEZZANOTTE (2009), ob. cit., nota 312, p. 121. Para ROLLA, “el decálogo al que sería preciso atenerse para que el derecho a la información no lesione el derecho a la vida privada puede encontrarse en la sentencia 5259/84 de la Corte de Casación, de acuerdo con la cual una información sería respetuosa con el derecho a la personal reserva cuando satisfaga los siguientes criterios: utilidad social de la información; verdad objetiva, incluyendo también en tal noción la veracidad o verdad putativa, fruto de un diligente y serio trabajo de contraste; forma adecuada de exposición y valoración de los hechos, excluida toda intención denigratoria”. Giancarlo ROLLA (2002), ob. cit., nota 305, p. 165.	10,212
https://openalex.org/W2045504350	OpenAlex	Open Science	CC-By	1,976	Generation of specific antiserum to thyrotropin releasing‐hormone and its use in a radioimmunoassay	Yitzhak Koch	English	Spoken	2,409	5,167	UC Irvine UC Irvine Previously Published Works Title Generation of specific antiserum to thyrotropin releasing‐hormone and its use in a radioimmunoassay Permalink https://escholarship.org/uc/item/8nf345t3 Journal FEBS Letters, 63(2) ISSN 0014-5793 Authors Koch, Y Baram, T Fridkin, M Publication Date 1976-04-01 DOI 10.1016/0014-5793(76)80115-9 Copyright Information This work is made available under the terms of a Creative Commons Attribution License, availalbe at https://creativecommons.org/licenses/by/4.0/ Peer reviewed UC Irvine UC Irvine Previously Published Works Title Generation of specific antiserum to thyrotropin releasing‐hormone and its use in a radioimmunoassay Permalink https://escholarship.org/uc/item/8nf345t3 Journal FEBS Letters, 63(2) ISSN 0014-5793 Authors Koch, Y Baram, T Fridkin, M Publication Date 1976-04-01 DOI 10.1016/0014-5793(76)80115-9 Copyright Information This work is made available under the terms of a Creative Commons Attribution License, availalbe at https://creativecommons.org/licenses/by/4.0/ Peer reviewed https://escholarship.org/uc/item/8nf345t3 Journal FEBS Letters, 63(2) Copyright Information This work is made available under the terms of a Creative Commons Attribution License, availalbe at https://creativecommons.org/licenses/by/4.0/ Peer reviewed Peer reviewed UC Irvine UC Irvine Previously Published Works Title Title Generation of specific antiserum to thyrotropin releasing‐hormone and its use in a radioimmunoassay * In partial fulfillment of the requirements for the Ph.D. degree of the Graduate School of the Weizmann Institute of Science. Powered by the California Digital Library University of California eScholarship.org eScholarship.org April 1976 April 1976 FEBS LETTERS Volume 63, number 2 1. Introduction TRH) was coupled to bovine serum albumin (BSA). p-Aminophenylacetic acid (0.094 mmol) in 0.4 ml of cold 2 N HC1 was diazotized by sodium nitrite (0.1 mmol in 50/~1 of water). Fifteen min later 0.094 mmol TRH (Calbiochem USA) in 0.4 rnl of 3 N KHCO3 was added. The reaction mixture, which gradually turned pinkish-red, was stirred for 10 h at 4°C. After acidification (pH 2.0) with 2 N HC1, the solution was extracted five times with ethylacetate, and the organic phase discarded; the aqueous phase was adjusted to pH 5.0 with KHCOa and lyophilized. The brownish product was extracted into absolute methanol, leaving insoluble white inorganic salts, and then precipitated by ether; the powder thus obtained was dissolved in absolute methanol and reprecipitated ether. Electrophoresis of the resultant brownish powder was carried out on Whatman No. 3 paper (pyridine acetate buffer, pH 6.5; 60 V/cm). Two components were detected with Pauli's reagent, one corresponding in mobility to unreacted TRH (30-50%, moving slightly towards the anode), the other (50-70%), moved towards the cathode. The latter component had a broad absorption peak in dimethyl- forinamide at 370-390 nm ()kma x = 380 nm; ~38o~DMF = 4.8 X 103), consistent with its identification as an azo-histidyl derivative [6,7]. Thyrotropin releasing-hormone (TRH), identified as the tripeptide pyroglutamyl-histidyl-proline amide, was isolated from hypothalamic tissue [ 1,2] and found to have both TSH and prolactin releasing activities [3]. Several reports described the genera- tion of antisera to the synthetic tripeptide after rendering it antigenic by coupling to a protein carrier, either through the proline residue [4] or through the histidine residue, using a symmetrical bifunctional coupling reagent in a one step reaction [5]. This paper describes the synthesis of an immunogenic protein conjugate of TRH using an asymmetrical bifunctional reagent in a two-step coupling procedure. Using antisera generated against this conjugate, a radioimmunoassay procedure for the peptide was developed, which permits the study of its distribu- tion in biological tissues. North.Holland Publishing Company - Amsterdam GENERATION OF SPECIFIC ANTISERUM TO THYROTROPIN RELEASING- HORMONE AND ITS USE IN A RADIOIMMUNOASSAY Y. KOCH, T. BARAM* Department of Hormone Research and M. FRIDKIN Department of Organic Chemistry The Weizmann Institute of Science, Rehovot, Israel M. FRIDKIN Department of Organic Chemistry The Weizmann Institute of Science, Rehovot, Israel Received 29 January 1976 2.3. Iodination and radioimmunoassay (RIA ) of TRH 2.3. Iodination and radioimmunoassay (RIA ) of TRH Labelling and RIA procedures for TRH were carried out as described previously for LH-Rtf [11]. Labelling and RIA procedures for TRH were carried out as described previously for LH-Rtf [11]. 3.3. Distribution of TRH The distribution of TRH in various regions of the rat brain is summarized in table 2. 2.2. Immunization The sensitivity of the radioimmunoassay for TRH was found to be approx. 10 pg (fig.l). A reference preparation of rat hypothalamic extract with TSH releasing activity (NIAMD-Rat HE-RP-1) inhibited the binding of [12Sl]TRIt by the antiserum to TRH- BSA. The inhibition curve was parallel to that pro- duced by synthetic TRH (fig. 1). The extent to which the antiserum to TRH cross-reacted with various analogues of TRH or other peptide hormones is summarized in table 1. Four mature male rabbits were injected subcutane- ously in several sites with an emulsion of TRH -BSA (0.25 rag/rabbit) in complete Freund's adjuvant supplemented with 1.6 units of a vaccine against Hemophilus pertussis (Pertussis Vaccine Fluid U.S.P., Eli Lilly & Co. Indianapolis). After two booster injections at intervals of two weeks, the rabbits were bled and the sera were examined for presence of antibodies. A conjugate of TRH and a multichain synthetic copolymer of L-lysine and DL-alanine: poly-DL-Ala- poly-L-lys, (tool. wt. = 142 500; Ala/Lys ratio = 20/I; [9] ) containing approximately 14% w/w of TRH was prepared by coupling the free carboxyl derivative of TRH (pGlu-His-Pro-OH) to the free amino groups of alanine, using N,N'-dicyclohexyl-carbodiimide in DMF as the coupling agent [ 10]. This preparation, when administered to rabbits, failed to produce anti TRH sera. 2.4. Dissection of various brain regions, and extraction o/ TRH Hy pot halamic extract(equivalents) ooz5oo5o~ o2 o5 too- I I I I I - c o Hyp0tholomic Z Q£ ~- 50 4 mH -~ ~-~ ~ TRH I I 1 I to too iooo 50oo TRH (pg) Fig. 1. Inhibition of binding of [ ~ 2 s I I TRH to antiserum by unlabelled TRH or by rat hypothalamic extract (NIAMD-Rat HE-RP-1). Anti-TRH serum used at 1:20 000 dilution. Hy pot halamic extract(equivalents) ooz5oo5o~ o2 o5 too- I I I I I - c o Hyp0tholomic Z Q£ ~- 50 4 mH -~ ~-~ ~ TRH I I 1 I to too iooo 50oo TRH (pg) Tissue fragments corresponding to different brain areas were excised according to de Groot [ 12]. TRH was extracted with 90% methanol, taken to dryness under a stream of air, and redissolved in 0.01 M phosphate-buffered saline (PBS), pH 6.9. 2.1. Preparation of TRH-BSA conjugate 2.1. Preparation of TRH-BSA conjugate p-Diazonium phenylacetic acid was attached to synthetic TRH and the resulting azo-derivative (PAPA- The PAPA-TRH derivative was attached to BSA (Crystalline grade, Miles Labs., Kankakee, Ill.), using 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide HC1 at pH 5.0 [8]. Spectral analysis of the TRH-BSA The PAPA-TRH derivative was attached to BSA (Crystalline grade, Miles Labs., Kankakee, Ill.), using 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide HC1 at pH 5.0 [8]. Spectral analysis of the TRH-BSA 295 North.Holland Publishing Company - Amsterdam FEBS LETTERS Volume 63, number 2 April 1976 ing titers of the sera from the other immunized rabbits ranged from 1 : 500 to 1 : 8000. conjugate at 380 nm indicated that on the average 24 residues of TRH were attached to each molecule of the carrier protein. ing titers of the sera from the other immunized rabbits ranged from 1 : 500 to 1 : 8000. 4. Discussion Two approaches have recently been used to render TRH antigenic: (i) Conjugation of TRH to BSA through the carboxy-terminal of pGlu-His- Pro-OH (TRH-OH) [4]. A radioimmunoassay system based on antibodies generated against this antigen were able to detect only high concentrations (above 100 pg) of TRH and cross-reacted extensively with 3.1. Formation of antibodies to TRH f Antibodies binding [12SI]TRH (~ 1 mCi//~g) appeared in the serum of the immunized rabbits two months after the first immunization. Three months later the serum of one immunized rabbit (No. 23) bound 50% of the labelled hormone (~ 50 pg per assay tube) at 1:20 000 final dilution; the correspond- Fig. 1. Inhibition of binding of [ ~ 2 s I I TRH to antiserum by unlabelled TRH or by rat hypothalamic extract (NIAMD-Rat HE-RP-1). Anti-TRH serum used at 1:20 000 dilution. 296 April 1976 April 1976 Volume 63, number 2 FEBS LETTERS Table 1 Specificity of anti-TRH serum Inhibition of No. Peptide [ 12~ I] TRH-binding (%) 1 pGlu His-Pro-NH: (TRH) 100 2 pGlu-His-Pro-OH (TRH-OH) 0.1 3 pGlu-His-Trp-OH 0.01 4 pGlu-His-OH 0.004 5 His-Pro-NH~ < 0.001 6 Pro-Leu-Gly-NH 2 (MIF) < 0.001 7 Vasopressin < 0.001 8 Oxytocin < 0.001 9 LHRH < 0.001 Peptides were tested in doses ranging from 10 ng to 100 ~g. Anti TRH was used at a final dilution of 1:20 000. Lysine-vasopressin and oxytocin were the product of Sigma Chemical Co., St. Louis. LH-RH was kindly provided by Hoechst, AG, Frankfurt. Compounds No. 3 and 4 were gifts from Dr W. F. White. Compounds No. 2, 5 and 6 were synthetized in our laboratory. Table 2 Distribution of TRH in rat brain Brain Region Wet weight (mg) TRH pg/mg Median eminence a 3.9 -+ 0.2 d 387.5 ± 21.2 Preoptic area 16.7 +- 1.8 92.7 -+ 9.8 Residual hypothalamus b 19.7 -+ 1.7 125.3 -+ 7.4 Cerebral cortex 39.6 -+ 4.9 0.3 +- 0.02 Posterior pituitary 1.1 +- 0.02 167.8 -+ 15 Amygdala 25.6 +- 1.1 6.1 ± 1.9 Hippocampus (dorsal) 45.4 ± 1.4 4.8 ± 0.4 Olfactory tubercle + anterior 47.4 ± 2.0 21.8 +- 3.5 olfactory nucleus Pineal gland c 5.0 ± 0.1 4.2 *- 0.1 a The 'median eminence' includes parts of the medial basal hypothalamic nuclei. b Excluding the preoptic area, the median eminence, and the mammillary bodies. c Pools of five glands. d Mean and standard error of the mean for 6-12 determinations. Table 2 Distribution of TRH in rat brain Distribution of TRH in rat brain a The 'median eminence' includes parts of the medial basal hypothalamic nuclei. utilizing a symmetrical bifunctional reagent can result in dimerization and oligomerization of TRH. We, therefore, used an asymmetrical bifunctional reagent in a two-step coupling procedure. References labelled TRH for antibodies binding sites when tested in a radioimmunoassay system (table 1). A rat hypo- thalamic extract (NIAMD-Rat HE-RP-1) with TSH- releasing activity displayed inhibition curves parallel to that of synthetic TRH (fig. 1). These characteristics, combined with high sensitivity (limit of detection approx. 10 pg), render this radioimmunoassay system capable of providing meaningful quantitative assess- ment of TRH in biological tissues. [1] Boler, J., Enzmann, F., Folkers, K., Bowers, C. Y. and Schally, A. V. (1969) Biochem. Biophys. Rcs. Commun. 37, 705-710. [2] Burgus, R., Dunn, T., Desiderio, D. and Guillemin, R. (1969) C. R. Acad. Sci., Paris 269, 1870-1872. [3] Vale, W., Blackwell, R., Grant, G. and Guillemin, R. (1973) Endocrinology 93, 26-33. (1973) Endocrinology 93, 26-33. ( ) gy [4] Visser, T. J., Docter, R. and Hennemann, G. (1974) Acta Endocrinol. 77, 417-421. Acta Endocrinol. 77, 417-421. The highest concentration of TRH in rat brain was found in the median eminence (387 pg]mg); high concentrations were also found in the rest of the hypothalamus (125 pg/mg) and in the posterior pituitary. These results are in accord with those obtained by others [ 15-17]. Lower concentrations were found in the preoptic area (93 pg/mg) and in the olfactory nucleus (22 pg/mg). In contrast to a recent report [18], only low TRtt levels (~ 4 pg/gland) were detected in the rat pineal gland. [5] Bassiri, R. M. and Utiger, R. D. (1972) Endocrinology 90, 722-727. [6] Cuatrecasas, P. (1970) J. Biol. Chem. 245, 574-584. [7] Nagai, W., Kirk, K. L. and Cohen, L. A. (1973) J. Org. Chem. 38, 1971-1972. [8] Goodfriend, T. L., Levine, L. and Fasman, G. D. (1964) Science 144, 1344-1346. [9] Sela, M., Fuchs, S. and Arnon, R. (1962) Biocbem. J. 85, 223-235. [10] Sheehan, J. C. and Hess, G. P. (1955) J. Am. Chem. Soc. 77, 1067-1068. [11] Koch, Y,, Wilchek, M., Fridkin, M., Chobsieng, P., Zor, U. and Lindner, H. R. (1973) Biochem. Biophys Res. Commun. 55,616-622. The high concentration of TRH in the rat hypo- thalamus is consistent with the biological role of this hormone in the regulation of TSH secretion from the pituitary. The extra-hypothalamic localization of TRH, however, may suggest further activities of this tripeptide, possibly exerted on neural tissue [19]. [ 12] de Groot, J. (1972) In: The Rat Forebrain in Stereo- toxic Coordinates, North-Holland Publishing Company, Amsterdam. [13] Nair, R. M. G., Redding, T. W. References and Schally, A. V. (1971) Biochemistry 10, 3621-3624. [14] Bryce, G. F. (1974) Immunoehemistry 11,507-511. [15] Jackson, I. M. D. and Reichlin, S. (1974) Endocrinology 95,854--862. 3.1. Formation of antibodies to TRH The antigen thus produced (PAPA-TRH) generated antisera with titers of up to 1:20 000, and showed negligible cross reaction with four unrelated hypothalamic hormones (<0.001%) or with closely related oligopeptides (table 1). Thus the TRH metabolite (pGlu-His-Pro- OH) showed little competition (0.1%) with intact utilizing a symmetrical bifunctional reagent can result in dimerization and oligomerization of TRH. We, therefore, used an asymmetrical bifunctional reagent in a two-step coupling procedure. The antigen thus produced (PAPA-TRH) generated antisera with titers of up to 1:20 000, and showed negligible cross reaction with four unrelated hypothalamic hormones (<0.001%) or with closely related oligopeptides (table 1). Thus the TRH metabolite (pGlu-His-Pro- OH) showed little competition (0.1%) with intact the pGlu-His-Pro-OMe and pGlu-His-Pro-OH analogues of TRH. Since TRH is rapidly inactivated in blood by conversion of the amide to a carboxyl group [13], results obtained with such antisera require careful interpretation. (ii) Coupling of TRH to BSA [5] or to thyroglobulin [14,15] via the histidyl moiety by the bis-diazotized benzidine technique. Visser et al. [4] however have failed to produce antiserum to TRH by this method. The reason for this may be that the one-step reaction 297 FEBS LETTERS April 1976 Volume 63, number 2 References Acknowledgements [16] Winokur, A. and Utiger, R. D. (1974) Science 185, 265-267. We are grateful to Miss N. Ben Aroya for excellent technical assistance, to Dr A. F. Parlow of the National Institute of Arthritis and Metabolic Diseases for reference preparation of rat hypothalamic extract and to Dr H. R. Lindner for the interest taken in this study and his assistance in the preparation of this manuscript. The work was supported by a grant (to H.R.L.) of the Ford Foundation and the Population Council Inc., N.Y. [17] Brownstein, M. J., Palkovits, M., Saavedra, J. M., Bassiri, R. M. and Utiger, R. D. (1974) Science 185, 267-269. [18] Montoya, E., Wilber, J., White, W. and Gendrich, R. (1975) in: Endocrine Society 57th Annual Meeting, New York, Abst. 88. [19] Mark, J. L. (1975) Science 190, 544-545. 298	30,314
https://openalex.org/W3015939620	OpenAlex	Open Science	CC-By	2,020	Management of central nervous system relapse in a young patient affected by primary mediastinal large B‐cell lymphoma: A case report	Miriam Marangon	English	Spoken	3,199	6,234	K E Y W O R D S autologous stem cell transplant, central nervous system involvement, MATRix regimen, primary mediastinal large B-cell lymphoma Abstract Abstract In primary mediastinal large B-cell lymphoma, central nervous system (CNS) re- lapse is an uncommon event with a dismal prognosis. We report about the success- ful management of CNS relapse with chemoimmunotherapy according to MATRix (methotrexate, cytarabine, thiotepa, and rituximab) protocol followed by autologous stem cell transplant. Correspondence p Pier Luigi Zinzani, Institute of Hematology “L. e A. Seràgnoli”, University of Bologna, Via Massarenti, 9-40138 Bologna, Italy. Email: pierluigi.zinzani@unibo.it Received: 2 July 2019 \| Revised: 24 October 2019 \| Accepted: 25 October 2019 Received: 2 July 2019 \| Revised: 24 October 2019 \| Accepted: 25 October 2019 Received: 2 July 2019 \| Revised: 24 October 2019 \| Accepted: 25 October 2019 DOI: 10.1002/ccr3.2706 DOI: 10.1002/ccr3.2706 Miriam Marangon \| Beatrice Casadei \| Alessandro Broccoli \| Lisa Argnani \| Michele Cavo \| Pier Luigi Zinzani Institute of Hematology “L. e A. Seràgnoli”, University of Bologna, Bologna, Italy Management of central nervous system relapse in a young patient affected by primary mediastinal large B-cell lymphoma: A case report Miriam Marangon \| Beatrice Casadei \| Alessandro Broccoli \| Lisa Argnani \| Michele Cavo \| Pier Luigi Zinzani This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2020 The Authors. Clinical Case Reports published by John Wiley & Sons Ltd. Clin Case Rep. 2020;8:933–937. \| 933 wileyonlinelibrary.com/journal/ccr3 This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2020 The Authors. Clinical Case Reports published by John Wiley & Sons Ltd. FIGURE 1 Magnetic resonance imaging at central nervous system lymphoma recurrence A 25-year-old young woman was transferred to our Institute in October 2016 from emergency department, where she was admitted due to the appearance of hacking cough, facial edema and conjunctival swelling. She had undergone a total body computed tomography (CT) scan, which showed a right mediastinal bulky mass (transverse diameters of 11 and 7.5 cm, respectively, and longitudinal diameter of 10 cm) compressing right cardiac structures, inferior vena cava, right pulmonary artery, right bronchus, surrounding ascending aorta and infiltrating pleura. Furthermore, supra and sub- diaphragmatic adenopathies, multiple pulmonary nodules, a right pleural effusion, pancreatic, renal and right adrenal involvement were observed. Subsequently, a CT-guided me- diastinal biopsy was performed, leading to a diagnosis of PMBCL. Immunophenotype on diagnostic biopsy was as follows: CD20+, PAX5+, CD30±, CD10-, BCL6+, BCL2±, IRF4+. 18F-fluorodeoxyglucose positron emission tomogra- phy (18FDG-PET) confirmed the presence of uptake at the mediastinal bulky mass, further mediastinal adenopathies, lungs, pancreas, kidneys, and right adrenal gland. Bone mar- row biopsy was negative. According to Ann Arbor staging system, the disease was stage IV A, due to the absence of lymphoma-related “B” symptoms. Physical examination re- vealed only the presence of neck and shoulders' edema; pe- ripheral blood count revealed mild anemia and a white blood cell count of 13.000/mmc, with a normal differential count; platelet count was normal. Lactate dehydrogenase (LDH) was elevated. Age-adjusted IPI was 2 (high-intermediate risk). FIGURE 1 Magnetic resonance imaging at central nervous system lymphoma recurrence scan and 18FDG-PET did not show further sites of disease involvement. Thus, the patient underwent partial avulsion of the le- sion through a left craniotomy at neurosurgical facility. Histological analysis revealed a diffuse large B-cell lym- phoma (DLBCL), which was considered to be consistent with a CNS recurrence of the primary mediastinal lymphoma. From October 2016 to January 2017, the patient was treated with 12 cycles of R-MACOP-B chemoimmunother- apy, which is administered weekly (cyclophosphamide and doxorubicin on cycles 1, 3, 5, 7, 9, 11; methotrexate and vincristine on cycles 2, 6, 10; bleomycin and vincristine on cycles 4, 8, 12) with prednisone given continuously and an- ti-CD20 rituximab boost during four of the 12 cycles. During the treatment, the patient experienced grade IV neutropenia responsive to granulocyte-colony stimulating factor, metho- trexate-related grade III oral mucositis and grade II nausea. 1 \| INTRODUCTION their third or fourth decades of life. It usually presents with a rapidly progressive mediastinal mass which can be associ- ated with symptoms related to superior vena cava syndrome. About 70% of patients have a bulky mass (>10 cm) at di- agnosis. Distant spread to extra-mediastinal lymph nodes or extranodal sites (ie, to the kidneys, adrenal glands, liver and ovaries) is uncommon at the time of presentation, but it can occur more frequently at the time of relapse.2,3 Central ner- vous system (CNS) involvement rarely occurs at the time of onset, and it can be a quite infrequent site of disease re- currence; the main pattern of CNS disease is constituted by parenchymal lesions.4,5 CNS recurrence is typically charac- terized by a dismal prognosis, and its treatment represents a difficult challenge for clinicians. Primary mediastinal large B-cell lymphoma (PMBCL) is an uncommon tumor which constitutes about 2%-3% of all non-Hodgkin lymphomas and 6%-10% of diffuse large B-cell lymphomas (DLBCL). In this disease, central nervous system (CNS) relapse is a quite uncommon event, which constitutes nevertheless a therapeutic challenge due to its dismal prog- nosis. Since in literature there are currently no data available concerning management of CNS relapse in PMBCL, we refer to studies conducted in DLBCL. Here, we report about the successful management of CNS relapse with chemoimmu- notherapy according to MATRix (methotrexate, cytarabine, thiotepa, and rituximab) protocol followed by autologous stem cell transplant in a young woman affected by PMBCL. Further follow-up is needed to determine long-term outcome. Here, we report about the successful management of CNS relapse with chemoimmunotherapy according to MATRix (methotrexate, cytarabine, thiotepa, and rituximab) proto- col followed by autologous stem cell transplant (ASCT) in a young patient affected by PMBCL.6,7 Primary mediastinal large B-cell lymphoma (PMBCL) is a mature B-cell neoplasm which arises from thymic B cells.1 This uncommon disease affects more often women in wileyonlinelibrary.com/journal/ccr3 933 Clin Case Rep. 2020;8:933–937. 934 MARANGON et al. A single, total dose of 2900 mg of cy- tarabine was delivered in the fourth cycle, before the develop- ment of hypersensitivity reaction. In a large trial conducted in prerituximab era involving 899 patients, a cumulative CNS relapse incidence of 2.8% was reported among patients affected by intermediate or high-grade lymphoma, who were treated with polichemo- therapy regimens. In this trial, CNS recurrence appeared to occur early (all cases occurred within 24 months from the end of treatment) and was characterized by a dismal prognosis.8 After fourth and last cycle of therapy, both MRI (Figure 2) and 18FDG-PET (Deauville score 1) showed a CR. Subsequently, the patient received conditioning treatment with carmustine and thiotepa, followed by ASCT. The main toxicities of this therapy were grade III oral and oropharyngeal mucositis associated with herpes simplex virus 1 reactivation, which required total paren- teral nutrition, antiacid, and antiviral treatment, and a prolonged hematologic toxicity. Engraftment of neutrophils (ANC > 500/ mmc) was observed on day +10 after ASCT, while engraftment of platelets (PLT count > 20.000/mmc) was observed on day +16. Both 18FDG-PET and CT scan of the brain showed a CR 2 months after ASCT. The addition of monoclonal antibody rituximab to poli- chemotherapy regimens such as CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) conferred an advantage in terms of risk of CNS involvement in DLBCL, although its real benefit is still controversial.9 Several attempts were made to better identify patients at high risk of CNS involvement, in order to guide specific diag- nostic workup and to select patients for appropriate therapeu- tic or prophylactic measures. Recently, an international group developed a risk model called “CNS-International Prognostic Index” (CNS-IPI).10 The score was developed analysing pa- tients who were enrolled in studies from the German High- Grade Non-Hodgkin Lymphoma Study Group and the MabThera International Trial and was subsequently validated in an independent data set from the British Columbia Cancer Lymphoid Cancer database. The study confirmed the infre- quence, earliness, and aggressive course of CNS relapse in aggressive B-cell lymphomas; the model defining the CNS- IPI included IPI factors. Due to the incidental finding of a thyroid nodule at 18FDG- PET scan, with normal thyroid function, an ultrasound and Doppler were performed. Ultrasound findings were consis- tent with the diagnosis of thyroid adenoma, which required annual follow-up. The patient was treated with 4 cycles of chemoimmuno- therapy with MATRix regimen.6 During the cycles, the patient experienced febrile epi- sodes with negative blood cultures, responsive to broad spec- trum antibacterial therapy, and an episode of blurred vision during the second cycle, with the finding of keratoconjuncti- vitis sicca at ophthalmic examination. After the second cycle, the patient presented a febrile episode associated with a chest CT scan finding consistent with invasive fungal infection; therefore, a broncho-alveolar lavage was performed, showing galactomannan positivity, and an antifungal therapy with am- photericin B was undertaken. During the 4 cycles of MATRix treatment, the patient developed grade IV neutropenia, which resolved after gran- ulocyte-colony stimulating factor (G-CSF) administration, grade III anemia, which required red blood cell transfusions, and grade IV thrombocytopenia, which required platelet transfusions. 18FDG-PET was performed after 6 cycles and after the completion of 12 cycles of therapy; at both time points, com- plete response (CR) was shown (Deauville score 1 and 1, respectively). In April 2017, 3 months after the completion of chemo- immunotherapy, the patient started complaining transient dysarthria and expressive aphasia; therefore, a cranial CT scan was immediately performed, showing a left temporal mass with associated edema. Magnetic resonance imaging (MRI) confirmed the finding (Figure 1), and a total body CT After the second cycle, an attempt to mobilize periph- eral blood stem cells (PBSC) was performed, without the obtainment of a PBSC count sufficient for collection; mo- bilizing treatment with granulocyte-colony stimulating fac- tor and plerixafor was therefore performed again after the third cycle, with the achievement of a CD 34+ cells'peak of MARANGON et al. 935 \| CNS relapse in PMBCL, we refer to studies conducted in DLBCL. 10.000/mL on day +19 after the completion of chemother- apy and subsequent collection of 3.54 × 106 CD34+ cells/kg. 10.000/mL on day +19 after the completion of chemother- apy and subsequent collection of 3.54 × 106 CD34+ cells/kg. The last cycle was complicated by a reaction to cytara- bine, characterized by fever, skin rash, and severe headache; the subsequent cytarabine and thiotepa administrations were therefore suspended. REFERENCES response rate, 30 months-progression-free survival and over- all survival, if compared to methotrexate and cytarabine or with methotrexate, cytarabine and rituximab in patients af- fected by primary CNS lymphoma. In this study, patients with responsive or stable disease after the first stage were randomised to receive whole-brain radiotherapy (WBRT) or ASCT; the results from the second randomisation showed that WBRT and ASCT are both feasible and effective as con- solidation therapies.6,7 1. Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016;127(20):2375-2390. 1. Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016;127(20):2375-2390. 2. Martelli M, Ferreri A, Di Rocco A, Ansuinelli M, Johnson PWM. Primary mediastinal large B-cell lymphoma. Crit Rev Oncol Hematol. 2017;113:318-327. 3. Zinzani PL, Piccaluga PP. Primary Mediastinal DLBCL: evolving biologic understanding and therapeutic strategies. Curr Oncol Rep. 2011;13(5):407-415. 4. Bishop PC, Wilson WH, Pearson D, Janik J, Jaffe ES, Elwood PC. CNS involvement in primary mediastinal large B-cell lymphoma. J Clin Oncol. 1999;17(8):2479-2485. Following the promising results showed in the study by Ferreri and colleagues, at the time of CNS relapse, we treated our patient with four courses of chemo-immunotherapy with methotrexate, cytarabine, thiotepa, and rituximab according to MATRix protocol, with the obtainment of a CR since the second cycle of treatment; although both ASCT and WBRT are effective consolidation strategies in patients affected by primary CNS DLBCL7, due to the young age of the pa- tient and the known risk of cognitive impairment related to WBRT, we decided to perform ASCT. 18FDG-PET and CT scan of the brain documented the maintenance of CR two months after the procedure. 5. Papageorgiou SG, Diamantopoulos P, Levidou G, et al. Isolated central nervous system relapses in primary mediastinal large B-cell lymphoma after CHOP-like chemotherapy with or without Rituximab. Hematol Oncol. 2013;31(1):10-17. 6. Ferreri AJM, Cwynarski K, Pulczynski E, et al. Chemoimmunotherapy with methotrexate, cytarabine, thiotepa, and rituximab (MATRix regimen) in patients with primary CNS lymphoma: results of the first randomisation of the International Extranodal Lymphoma Study Group-32 (IELSG32) phase 2 trial. Lancet Haematol. 2016;3(5):e217-227. 7. Ferreri AJM, Cwynarski K, Pulczynski E, et al. Of note, in the validation study, pa- tients affected by PMBCL were excluded; another limitation of the study is the fact that, even with this robust risk score, a very high-risk group remains difficult to identify.10 At di- agnosis, our patient had kidney and adrenal gland involve- ment, more than one extranodal site, stage IV disease and elevated LDH; therefore, she had high-risk disease (CNS-IPI 4). Due to the lack of consensus and of recommendations on CNS prophylaxis in PMBCL and the failure of several stud- ies to demonstrate a benefit of intrathecal (IT) prophylaxis in DLBCL, we did not perform IT prophylaxis after first-line chemoimmunotherapy.11,12 The case report study was approved by the local ethical committee. Written informed consent for publication of her clinical details and clinical images was obtained from the pa- tient. Timeline of the case report is shown in Table 1. PMBCL is an uncommon tumor which constitutes about 2%-3% of all non-Hodgkin lymphomas and 6%-10% of DLBCL.2 In this disease, CNS relapse is a quite uncommon event, which constitutes nevertheless a therapeutic chal- lenge due to its dismal prognosis. Since in literature there are currently no data available concerning management of FIGURE 2 Magnetic resonance imaging at the end of MATRix (methotrexate, cytarabine, thiotepa, and rituximab) regimen (complete response) Since the patient achieved a complete remission after first- line treatment, we did not perform consolidative mediastinal radiotherapy (RT); in fact, as recent studies suggested that consolidation with RT can be safely omitted in patients who achieve PET-negativity after first-line chemoimmunotherapy, we do not usually administer RT in this setting of patients.13-15 Patients who experience CNS relapse have a poor progno- sis; thus, they should receive treatment according to modern protocols for primary CNS lymphoma, such as methotrex- ate-/cytarabine-based schemes. Recently, in an international randomised phase II trial, the addition of rituximab and thiotepa to a cytarabine and methotrexate therapeutic scheme showed a significant benefit in terms of CR rate, overall FIGURE 2 Magnetic resonance imaging at the end of MATRix (methotrexate, cytarabine, thiotepa, and rituximab) regimen (complete response) 936 \| October 2016 Diagnosis of PMBCL October 2016 R-MACOP-B CR April 2017 CNS recurrence April 2017 Start of MATRix regimen June 2017 Mobilization of peripheral blood stem cells August 2017 End of MATRix regimen CR January 2018 ASCT March 2018 Evaluation post-ASCT CR Abbreviations: ASCT, autologous stem cell transplant; CNS, central nervous system; CR, complete response; MATRix, methotrexate, cytarabine, thiotepa, and rituximab; PMBCL, primary mediastinal B-cell lymphoma; R-MACOP-B, rituximab, methotrexate, doxorubicin, cyclophosphamide, vincristine, prednisone, bleomycin. 936 MARANGON et al. MARANGON et al. 936 \| MARANGON et al. October 2016 Diagnosis of PMBCL October 2016 R-MACOP-B CR April 2017 CNS recurrence April 2017 Start of MATRix regimen June 2017 Mobilization of peripheral blood stem cells August 2017 End of MATRix regimen CR January 2018 ASCT March 2018 Evaluation post-ASCT CR Abbreviations: ASCT, autologous stem cell transplant; CNS, central nervous system; CR, complete response; MATRix, methotrexate, cytarabine, thiotepa, and rituximab; PMBCL, primary mediastinal B-cell lymphoma; R-MACOP-B, rituximab, methotrexate, doxorubicin, cyclophosphamide, vincristine, prednisone, bleomycin. TABLE 1 Timeline Abbreviations: ASCT, autologous stem cell transplant; CNS, central nervous system; CR, complete response; MATRix, methotrexate, cytarabine, thiotepa, and rituximab; PMBCL, primary mediastinal B-cell lymphoma; R-MACOP-B, rituximab, methotrexate, doxorubicin, cyclophosphamide, vincristine, prednisone, bleomycin. CONFLICT OF INTEREST 8. Bernstein SH, Unger JM, LeBlanc M, Friedberg J, Miller TP, Fisher RJ. Natural history of CNS relapse in patients with ag- gressive non Hodgkin’s lymphoma: a 20-year follow-up analysis of SWOG 8516-The Southwest Oncology Group. J Clin Oncol. 2009;27(1):114-119. None declared. AUTHOR CONTRIBUTIONS MM, BC, AB, and PLZ: analyzed and interpreted the pa- tient data regarding the hematologic disease; MC: pro- vided advice for treatment and analyzed and interpreted the patient data regarding the hematologic disease; MM, LA, and PLZ: were major contributor in writing the man- uscript; and all authors: read, revised, and approved the final manuscript. 9. Villa D, Connors JM, Shenkier TN, Gascoyne RD, Sehn LH, Savage KJ. Incidence and risk factors for central nervous system relapse in patients with diffuse large B cell lymphoma: The impact of the addition of rituximab to CHOP chemotherapy. Ann Oncol. 2010;21(5):1046-1052. 10. Schmitz N, Zeynalova S, Nickelsen M, et al. CNS International prognostic index: a risk model for CNS relapse in patients with diffuse large B-cell lymphoma treated with R-CHOP. J Clin Oncol. 2016;34(26):3150-3156. REFERENCES Whole-brain radio- therapy or autologous stem-cell transplantation as consolidation strategies after high-dose methotrexate-based chemoimmunother- apy in patients with primary CNS lymphoma: results of the second randomisation of the International Extranodal Lymphoma Study Group-32 phase 2 trial. Lancet Haematol. 2017;4(11):e510-e523. In conclusion, we report the successful management of a CNS relapse of PMBCL with MATRix treatment; further follow-up is needed to determine long-term outcome. ORCID ORCID Pier Luigi Zinzani https://orcid.org/0000-0002-2112-2651 937 MARANGON et al. \| 15. Dunleavy K, Pittaluga S, Maeda LS, et al. Dose adjusted EPOCH- rituximab therapy in primary mediastinal B-cell lymphoma. N Engl J Med. 2013;368:1408-1416. 15. Dunleavy K, Pittaluga S, Maeda LS, et al. Dose adjusted EPOCH- rituximab therapy in primary mediastinal B-cell lymphoma. N Engl J Med. 2013;368:1408-1416. 11. Kumar A, Vanderplas A, LaCasce AS, et al. Lack of benefit of central nervous system prophylaxis for diffuse large B-cell lym- phoma in the rituximab era: findings from a large national data- base. Cancer. 2012;118:2944-2951. 12. Guirguis HR, Cheung MC, Mahrous M, et al. Impact of central nervous system (CNS) prophylaxis on the incidence and risk fac- tors for CNS relapse in patients with diffuse large B-cell lymphoma treated in the rituximab era: a single centre experience and review of the literature. Br J Haematol. 2012;159:39-49. How to cite this article: Marangon M, Casadei B, Broccoli A, Argnani L, Cavo M, Zinzani PL. Management of central nervous system relapse in a young patient affected by primary mediastinal large B-cell lymphoma: A case report. Clin Case Rep. 2020;8:933–937. https://doi.org/10.1002/ccr3.2706 How to cite this article: Marangon M, Casadei B, 13. Broccoli A, Casadei B, Stefoni V, et al. The treatment of primary mediastinal large B-cell lymphoma: a two decades monocentric ex- perience with 98 patients. BMC Cancer. 2017;17:276. 14. Zinzani PL, Broccoli A, Casadei B, et al. The role of rituximab and positron emission tomography in the treatment of primary medias- tinal large B-cell lymphoma: experience on 74 patients. Hematol Oncol. 2015;33(4):145-150. 14. Zinzani PL, Broccoli A, Casadei B, et al. The role of rituximab and positron emission tomography in the treatment of primary medias- tinal large B-cell lymphoma: experience on 74 patients. Hematol Oncol. 2015;33(4):145-150.	8,010
https://openalex.org/W4360868623	OpenAlex	Open Science	CC-By	2,023	A novel zinc metabolism-related gene signature to predict prognosis and immunotherapy response in lung adenocarcinoma	Wuguang Chang	English	Spoken	6,359	11,562	TYPE Original Research PUBLISHED 24 March 2023 DOI 10.3389/fimmu.2023.1147528 OPEN ACCESS OPEN ACCESS EDITED BY Parmanand Malvi, University of Alabama at Birmingham, United States REVIEWED BY Remya Nair, Winship Cancer Institute, Emory University, United States Subhash Kumar Tripathi, Seattle Children’s Research Institute, United States CORRESPONDENCE Si-Yu Wang wangsy@sysucc.org.cn Wei Ou ouwei@sysucc.org.cn †These authors share ﬁrst authorship SPECIALTY SECTION This article was submitted to Cancer Immunity and Immunotherapy, a section of the journal Frontiers in Immunology RECEIVED 18 January 2023 ACCEPTED 15 March 2023 PUBLISHED 24 March 2023 CITATION Chang W, Li H, Ou W and Wang S-Y (2023) A novel zinc metabolism-related gene signature to predict prognosis and immunotherapy response in lung adenocarcinoma. Front. Immunol. 14:1147528. doi: 10.3389/fimmu.2023.1147528 COPYRIGHT © 2023 Chang, Li, Ou and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that EDITED BY Parmanand Malvi, University of Alabama at Birmingham, United States REVIEWED BY Remya Nair, Winship Cancer Institute, Emory University, United States Subhash Kumar Tripathi, Seattle Children’s Research Institute, United States Wuguang Chang1,2†, Hongmu Li1,2†, Wei Ou1,2 and Si-Yu Wang1,2* 1State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China, 2Department of Thoracic Surgery, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong, China Background: Zinc is a key mineral element in regulating cell growth, development, and immune system. We constructed the zinc metabolism- related gene signature to predict prognosis and immunotherapy response for lung adenocarcinoma (LUAD). Methods: Zinc metabolism-associated gene sets were obtained from Molecular Signature Database. Then, the zinc metabolism-related gene signature (ZMRGS) was constructed and validated. After combining with clinical characteristics, the nomogram for practical application was constructed. The differences in biological pathways, immune molecules, and tumor microenvironment (TME) between the different groups were analyzed. Tumor Immune Dysfunction and Exclusion algorithm (TIDE) and two immunotherapy datasets were used to evaluate the immunotherapy response. Results: The signature was constructed according to six key zinc metabolism- related genes, which can well predict the prognosis of LUAD patients. The nomogram also showed excellent prediction performance. Functional analysis showed that the low-risk group was in the status of immune activation. More importantly, the lower risk score of LUAD patients showed a higher response rate to immunotherapy. COPYRIGHT © 2023 Chang, Li, Ou and Wang. COPYRIGHT © 2023 Chang, Li, Ou and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. KEYWORDS 1 Introduction than 30 days. Finally, we collected 485 LUAD and 59 normal samples. In addition, we also collected the copy number variation (CNV) data of LUAD in the TCGA database for analysis. The GSE72094 containing 398 LUAD from GEO was used as the validation set. Zinc metabolism-related genes were gathered from the molecular signature database (18). Lung cancer is the thorniest cancer in the world, and LUAD is the most prevalent type of non-small cell lung cancer (NSCLC), accounting for about 40% of lung cancer (1). In the past decades, with the increasing understanding of the molecular level, the treatment of lung cancer has undergone profound changes. Surgery or radiation is still the primary treatment for the early stage of lung cancer, while patients with advanced lung cancer have a variety of treatment options, including targeted treatment, immunotherapy, and combined with other therapies (2). Among them, immunotherapy brings great beneﬁts to patients with advanced lung cancer. However, due to the heterogeneity of tumors in different individuals, about 70% of patients with advanced non-small cell lung cancer did not respond to immunotherapy (3). Therefore, the search for new targets still needs to be continued. 2.2 Identiﬁcation of differently expressed zinc metabolism-related genes and enrichment analysis The differentially expressed zinc metabolism-related genes between tumor and normal tissues in the TCGA dataset were analyzed by ‘limma’ package (19). The screening criteria were set as adjusted p value < 0.05 and log2 \|Fold Change\| ≥1, which were displayed by heatmap and volcano plot. Subsequently, GO/KEGG enrichment analysis was performed on these genes through the ‘clusterProﬁler’ package (20). The ‘CNBplot’ package is a tool for Bayesian network inference of enrichment analysis results (21). We used it to infer the gene interaction of key pathways. Zinc is one of the essential trace elements in the body (4). It is a key component of a variety of enzymes and transcription factors, including oxidoreductase, hydrolase, lyase, synthetase, and ligase, which are involved in regulating DNA synthesis and cell cycle (5, 6). Therefore, the lack of zinc will lead to the disorder of DNA replication process, and then cause the cell to lose control of proliferation and become cancerous. The zinc content in the serum of various cancer patients has been signiﬁcantly reduced, including breast cancer (7), lung cancer (8), head and neck cancer (9), and prostate cancer (10). In addition, zinc widely affects the immune system of the body (11). Yu et al. found that the inﬂux of zinc increased after T cell receptor (TCR) was initially stimulated, and then accelerated the activation of T cell by lowering the threshold of TCR activation, thereby enhancing the immune response (12). Various zinc ﬁnger proteins participate in the regulation of B cell maturation (13, 14). The deﬁciency of zinc transporter SLC39A10 will cause the decrease of zinc level in macrophages, which leads to p53 mediated apoptosis (15). Zinc deﬁciency also promotes the production of inﬂammatory cytokines IL-1b, IL-6, and TNF-a via the MAPK pathway (16). In view of the extensive role of metals such as iron and copper in cancer and the cell death they cause (ferroptosis and cuproptosis) (17), we believe that regulating the content of zinc in the body and targeting the key enzymes in zinc metabolism are expected to become new targets for cancer treatment in the future. OPEN ACCESS This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Conclusion: The state of zinc metabolism is closely connected to prognosis, tumor microenvironment, and response to immunotherapy. The zinc metabolism-related signature can well evaluate the prognosis and immunotherapy response for LUAD patients. zinc, zinc metabolism, biomarker, prognosis, lung adenocarcinoma, immunotherapy 01 Frontiers in Immunology frontiersin.org Chang et al. 10.3389/ﬁmmu.2023.1147528 2.4 Nomogram for clinical application Through cox regression analysis, we explored whether clinical factors affected risk score, and the results were shown by forest plot. To establish a clinically feasible scoring system, based on the results of cox regression analysis, we combined the risk score and the clinical information (p < 0.05) and constructed the nomogram. The reliability of the nomogram was veriﬁed by receiver operating characteristic (ROC) curve and calibration curve. In this study, we constructed ZMRGS, which can accurately predict the prognosis of LUAD patients. It can also well evaluate the immunological characteristics and immunotherapy response of different LUAD patients. 2.3 Construction and validation of the zinc metabolism-related gene signature To build a reliable prognostic signature, we ﬁrst screened prognostic genes through univariate cox regression analysis and then incorporated them into the LASSO cox regression model to further reduce the number of candidate genes. Finally, the ZMRGS was constructed by multivariate cox regression analysis. The risk score of each LUAD sample = Coefgene1Expgene1+Coefgene2Expgene2 … CoefgenenExpgenen. According to the median risk score, LUAD patients were divided into high- and low-risk groups. KM survival analysis was used to analyze the survival differences between the two groups. Then the risk score of each LUAD patient in GSE72094 was calculated using the same algorithm and the same analysis was performed. Frontiers in Immunology 3.1 Differently expressed Zinc metabolism-related genes and function enrichment The ﬂow chart of this study was shown in Figure 1. By analyzing the transcriptome differences between LUAD and normal tissues, we obtained 33 differentially expressed zinc metabolism-related genes (Figure 2A), of which 11 were up-regulated and 22 were down-regulated (Figure 2B). Based on the TCGA database, 31.6% (176/557) of the samples had gene mutations (Figure 2C), of which CPS1 mutation rate was the highest, reaching 11%. In addition, the 2.1 Data collection and processing With the purpose of ﬁnding out the reasons that affect the survival differences between two risk groups, we used GSEA software (version: 4.2.3) to study the biological functions and Transcriptome data of LUAD from TCGA was used for the training set. To reduce the impact of non-tumor factors, we excluded the lack of survival data and the samples with OS less Frontiers in Immunology 02 frontiersin.org Chang et al. 10.3389/ﬁmmu.2023.1147528 pathways of the two subgroups. The threshold is set to p < 0.05 and false discovery rate (FDR) < 0.25. Shanghai, China) was used to amplify the target gene. Repeat the qRT-PCR assays for three times in 10ml reaction volume for each sample. The PCR primers used for ampliﬁcation were as follows: ABCC8, 5′- TCACCTCCGTGGTCTACTATC -3′ (forward),5′- CTTGGTCTGTATTGCTCCTCTC -3′ (reverse); CPS1, 5′- CAA CCTGGCAGTTCCTCTATAC-3′ (forward), 5′- ACAGCG TCCATTTCTACTTCTC -3′ (reverse); HMGA2, 5′- CAGGAAG CAGCAG-CAAGAA -3′ (forward),5′- CCAGGCAAGGCAACAT TGA -3′ (reverse); HVCN1, 5′- TGCCTGGAACATCAACTACAA -3′ (forward), 5′- CTCCAGGCGGAAGACAAATAA -3′ (reverse); MT1A, 5′- CGCCTTATAGCCTCTCAACTTC -3′ (forward),5′- TAAATGGGTCAGGGTTGTATGG -3′ (reverse); SLC39A11, 5′- CAGCTCTCGTGTTCGTATTCTC -3′ (forward), 5′- AGGAT GCCAGTTTCCCATTAC -3′ (reverse); GAPDH, 5′-GA TTCCACCCATGGCAAATTC-3′ (forward), 5′- GTCATGAG-TC CTTCCACGATAC -3′ (reverse). By the comparative threshold cycle (2-Ct) method, we calculated the relative expression of these 6 genes by paired test. 2.6 Analysis of tumor environment features Further to explore the differences in the characteristics of immune molecules in different subgroups, we extensively analyzed a variety of immune molecules, including chemokine, immunostimulator, immunoinhibitor, MHC molecule, and receptor. The correlation between key immune checkpoints and risk score was analyzed by spearman correlation analysis. Then, we used ssGSEA to analyze the composition of 28 kinds of immune cells in the TME (22). The ‘estimate’ package was used to analyze the differences of tumor microenvironment components as a whole (23). 2.10 Statistical analysis The TIDE algorithm is a method to calculate the dysfunction and exclusion of patients to immunotherapy according to the gene expression proﬁle (24). The higher TIDE score means that patients are more prone to immune escape when receiving immunotherapy. We calculated the TIDE score of each LUAD patient and analyzed the response of different subgroups to immunotherapy through chi- square test. In order to further verify the predictive efﬁcacy of zinc metabolism-related gene signature, we used two immunotherapy datasets with complete transcriptome data to verify (1) IMvigor210: a cohort of 298 patients with advanced urothelial carcinoma undergoing anti-PD-L1 (atezolizumab) immunotherapy (25). (2) Checkmate: a cohort of 181 patients with advanced renal cell carcinoma undergoing anti-PD-1 (nivolumab) immunotherapy (26). All graphs and data analysis were conducted by R software (version: 4.13) and SPSS (version: 26). Chi-square test was used for categorical variables. The continuous variable was used by the Wilcoxon test. 2.8 Analysis of drug sensitivity Based on the Cancer Genome Project (CGP) 2016 data in the ‘pRRophetic’ package (27), we analyzed the sensitivity of several commonly used chemotherapy drugs and compared the differences between the two groups. 2.9 Quantitative real-time PCR (qRT-PCR) FIGURE 1 Flowchart in this study. FIGURE 1 Flowchart in this study. The expression proﬁles of 6 key genes were veriﬁed by quantitative real-time PCR (qRT-PCR). There were 16 paired LUAD and lung tissues used for experiment, which were from patients undergoing lung cancer resection at our center. This study was approved by the Ethics Committee of Sun Yat-sen University Cancer Center (YB2018-85). A TRIzol (TIANGEN, Beijing, China) was used to isolate total RNA from cancer tissue samples and adjacent normal tissue samples. PrimeScriptTM RT Master Mix (ES Science, Shanghai, China) was used to reverse-transcribe complementary DNA. SYBR Green Master Mix (ES Science, Frontiers in Immunology 03 frontiersin.org Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D E F G A H I FIGURE 2 Characteristic of zinc metabolism-related genes in LUAD. (A) Heatmap showing the differences of zinc metabolism-related genes in LUAD and normal samples. (B) Volcano plot exhibiting 22 down-regulated and 11 up-regulated genes. (C) Gene mutation landscape of LUAD in TCGA. (D) The CNV mutation frequency of 33 zinc metabolism-related genes. (E) Chromosome position and alteration of zinc metabolism-related genes. (F) GO enrichment analysis. (G) KEGG enrichment analysis. Gene interaction network diagram in zinc ion homeostasis (H) and response to zinc ion (I). C E D B E D E D B F H F G I H H I FIGURE 2 Characteristic of zinc metabolism-related genes in LUAD. (A) Heatmap showing the differences of zinc metabolism-related genes in LUAD and normal samples. (B) Volcano plot exhibiting 22 down-regulated and 11 up-regulated genes. (C) Gene mutation landscape of LUAD in TCGA. (D) The CNV mutation frequency of 33 zinc metabolism-related genes. (E) Chromosome position and alteration of zinc metabolism-related genes. (F) GO enrichment analysis. (G) KEGG enrichment analysis. Gene interaction network diagram in zinc ion homeostasis (H) and response to zinc ion (I). 3.2 Construction and validation of the zinc metabolism-related gene signature proportion of CNV gain of most genes was higher, only the results of GATA1 and OTC were opposite (Figure 2D). The position of 33 genes on the chromosome was shown in Figure 2E. GO/KEGG enrichment analysis results showed that these genes were mainly concentrated in zinc metabolism activities, zinc homeostasis, and metabolism of various nutrients (Figures 2F, G). Then, we conducted the Bayesian network analysis on the key zinc metabolism pathway (zinc homeostasis and response to zinc) and showed the interaction between genes (Figures 2H, I). Univariate cox regression found that 9 genes were associated with prognosis (Figure S1). Based on the minimal lambda value of LASSO regression, we identiﬁed 8 genes (Figures 3A, B). Then, six key genes and their coefﬁcients were obtained through multivariate cox analysis (Figure 3C). Risk score = -ABCC80.407001 +CPS10.105130+HMGA20.192565-HVCN10.300203+MT1A Frontiers in Immunology frontiersin.org 04 Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D E F G H I J A FIGURE 3 Construction and validation of ZMRGS. (A) Tenfold cross-validation in LASSO model. (B) LASSO coefﬁcients of 9 prognostic-related genes. (C) 6 key zinc metabolism-related genes and their coefﬁcients. (D) Kaplan-Meier survival analysis in TCGA cohort. (E) Distribution of risk score and OS status in TCGA cohort. (F) Heatmap of 6 zinc metabolism-related genes in TCGA cohort. (G) Kaplan-Meier survival analysis in GSE72094. (H) Distribution of risk score and OS status in GSE72094. (I) Heatmap of 6 zinc metabolism-related genes in GSE72094. (J) The mRNA expressions of ABCC8, CPS1, HMGA2, HVCN1, MT1A and SLC39A11 by qRT-PCR. p < 0.05, *p < 0.01. C B A A C B E D D E F D E F F G H I H I G H J J Construction and validation of ZMRGS. (A) Tenfold cross-validation in LASSO model. (B) LASSO coefﬁcients of 9 prognostic-related genes. (C) 6 key zinc metabolism-related genes and their coefﬁcients. (D) Kaplan-Meier survival analysis in TCGA cohort. (E) Distribution of risk score and OS status in TCGA cohort. (F) Heatmap of 6 zinc metabolism-related genes in TCGA cohort. (G) Kaplan-Meier survival analysis in GSE72094. (H) Distribution of risk score and OS status in GSE72094. (I) Heatmap of 6 zinc metabolism-related genes in GSE72094. (J) The mRNA expressions of ABCC8, CPS1, HMGA2, HVCN1, MT1A and SLC39A11 by qRT-PCR. p < 0.05, *p < 0.01. 3.5 Immune molecule analysis To investigate the differences of immune molecules between different subgroups, we systematically analyzed the differences between chemokine, immunostimulator, immunoinhibitor, MHC molecule, and receptor (Figures 6A–E). There were obvious differences in the expression of most immune molecules, and almost all immune molecules were more abundant in the low-risk group. Subsequently, we also analyzed the correlation between several key immune checkpoints and the risk score. The results 3.3 Establishment of the nomogram 0.123295+SLC39A110.207582. KM survival analysis showed that the prognosis of low-risk group was better (p = 0.00014, Figure 3D). The distribution of risk scores showed that more people in high-risk groups died (Figure 3E). Figure 3F showed the expression of six key genes. In addition, similar results were obtained in GSE72094 (Figures 3G–I). The qRT-PCR showed that a high expression of CPS1, HMGA2, and SLC39A11 was found in tumor compared with adjacent normal tissues by 16 paired LUAD samples from our center, while the expression of ABCC8, HVCN1, and MT1A was lower in tumor (Figure 3J). Univariate and multivariate cox regression analysis all showed that risk score was an independent risk factor affecting the prognosis of LUAD (Figures 4A, B). Then, based on the above results, we built the nomogram to provide the basis for clinical practice (Figure 4C). The area under curve (AUC) values in 1, 3, and 5 years were 0.793, 0.741, and 0.722 respectively (Figure 4D). The correction curve also showed that the predicted value was consistent with the actual results (Figure 4E). Frontiers in Immunology 05 frontiersin.org Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D E A FIGURE 4 Construction of the nomogram to predict the prognosis of LUAD. Univariate (A) and multivariate (B) Cox regression analysis of risk score. (C) Nomogram for the prediction of 1-, 3- and 5-year survival probability. (D) Time-dependent ROC analysis of the nomogram. (E) Calibration curves for evaluating the accuracy. p < 0.01, p < 0.001. A B D D E C C E E FIGURE 4 Construction of the nomogram to predict the prognosis of LUAD. Univariate (A) and multivariate (B) Cox regression analysis of risk score. (C) Nomogram for the prediction of 1-, 3- and 5-year survival probability. (D) Time-dependent ROC analysis of the nomogram. (E) Calibration curves for evaluating the accuracy. p < 0.01, **p < 0.001. 3.4 Potential functions and pathways in different subgroups showed that except CD276, the other immune checkpoints were negatively correlated with risk score, while the immune checkpoints were positively correlated (Figure 6F). Based on GSEA, we analyzed the functional and pathway differences between different subgroups. GO enrichment analysis showed that there were a large number of pathways related to cell proliferation and cell cycle regulation in high-risk group (Figure 5A). In the low-risk group, a large number of immune- related pathways were activated (Figure 5B). The results of KEGG enrichment analysis also conﬁrmed the above conclusions (Figures 5C, D). 3.6 Differences of the tumor environment Based on ssGSEA algorithm, we found that up to 16 kinds of immune cells were signiﬁcantly inﬁltrated in the low-risk group (57.1%), while only CD4 T cell and type 2 T helper cell were signiﬁcantly inﬁltrated in the high-risk group (7.14%) (Figure 7A). The ESTIMATE algorithm showed that the low-risk group had a higher immune score (Figure 7B) and a lower tumor purity (Figure 7C). However, there was no signiﬁcant difference between the stromal score (Figure 7D). 3.8 Analysis of drug sensitivity According to the data from Cancer Genome Project (CCP) 2016 in the ‘pRRophetic’ package, we analyzed the relative sensitivity of various drugs. The overall drug sensitivity analysis results were shown in Figure S2. Surprisingly, the high-risk group was more sensitive to common drugs (Figures 8A–H). 3.7 Prediction of immunotherapy response by TIDE and immunotherapy datasets The prognostic signature of this study was composed of six genes (ABCC8, CPS1, HMGA2, HVCN1, MT1A, and SLC39A11). Previous studies had described the partial relationship between these genes and tumors. ABCC8 is a member of the MRP family involved in multidrug resistance. It was regarded as a prognostic marker for glioma and could predict chemosensitivity (29). CPS1 was thought to be involved in metabolic reprogramming in hepatocellular carcinoma, thus affecting the occurrence of tumors (30). The development of inhibitors for CPS1 also shows potential therapeutic potential (31). The high expression of HMGA2 in lung cancer indicates a worse prognosis, which is consistent with our ﬁndings. Vivo experiments have conﬁrmed that overexpression of HMGA2 can inhibit the proliferation and metastasis of lung cancer, so HMGA2 is expected to become a new target for the treatment of lung cancer (32). HVCN1 is the only gene encoding mammalian proton channel. Zinc ion can promote cell apoptosis and inhibit the invasion and metastasis of glioma cells by inhibiting the activity of HVCN1 in vivo and in vitro. MT1A is involved in DNA damage reaction and metal homeostasis and is therefore inseparable from the occurrence and development of tumors (33). SLC39A11 is a zinc transporter, which can inhibit the cloning of LUAD cells after being knocked down in vitro (34). result of response to immunotherapy also showed that the low-risk group was more likely to be responsive to immunotherapy (chi- square test, p < 0.0001, Figure 7F). In addition, we conducted KM survival analysis by using two independent immunotherapy datasets, and the results showed that the prognosis of high-risk patients was worse (IMvigor210: p = 0.0079, Checkmate: p = 0.003, Figures 7G, H). 3.7 Prediction of immunotherapy response by TIDE and immunotherapy datasets The results of TIDE suggested that the high-risk group had the higher TIDE score and exclusion score, while the low-risk group had higher dysfunctional score (Figure 7E), which meant that the high-risk group patients were more prone to immune escape. The Frontiers in Immunology 06 frontiersin.org Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D A FIGURE 5 Gene set enrichment analysis between two ZMRGS groups. GO enrichment in high-risk group (A) and low-risk group (B). KEGG enrichment in high- risk group (C) and low-risk group (D). B A B C D C FIGURE 5 Gene set enrichment analysis between two ZMRGS groups. GO enrichment in high-risk group (A) and low-risk group (B). KEGG enrichment in high- risk group (C) and low-risk group (D). The signature showed superior prediction ability. To get closer to clinical practice, we combined the signature with TNM staging to construct the nomogram. The prognostic signature of this study was composed of six genes (ABCC8, CPS1, HMGA2, HVCN1, MT1A, and SLC39A11). Previous studies had described the partial relationship between these genes and tumors. ABCC8 is a member of the MRP family involved in multidrug resistance. It was regarded as a prognostic marker for glioma and could predict chemosensitivity (29). CPS1 was thought to be involved in metabolic reprogramming in hepatocellular carcinoma, thus affecting the occurrence of tumors (30). The development of inhibitors for CPS1 also shows potential therapeutic potential (31). The high expression of HMGA2 in lung cancer indicates a worse prognosis, which is consistent with our ﬁndings. Vivo experiments have conﬁrmed that overexpression of HMGA2 can inhibit the proliferation and metastasis of lung cancer, so HMGA2 is expected to become a new target for the treatment of lung cancer (32). HVCN1 is the only gene encoding mammalian proton channel. Zinc ion can promote cell apoptosis and inhibit the invasion and metastasis of glioma cells by inhibiting the activity of HVCN1 in vivo and in vitro. MT1A is involved in DNA damage reaction and metal homeostasis and is therefore inseparable from the occurrence and development of tumors (33). SLC39A11 is a zinc transporter, which can inhibit the cloning of LUAD cells after being knocked down in vitro (34). The signature showed superior prediction ability. To get closer to clinical practice, we combined the signature with TNM staging to construct the nomogram. Frontiers in Immunology 4 Discussion Research on the connection between zinc and cancer has made some hopeful strides in recent years. Zinc widely affects the proliferation of tumor cells and the function of immune system (28), more and more studies have proved that zinc plays an irreplaceable role in the occurrence and development of tumors. However, the study of key genes in zinc metabolism is rare. As far as we know, this is the ﬁrst study to analyze the prognostic value and immune characteristics of zinc metabolism-related genes in LUAD. Zinc is involved in the formation of various enzymes that regulate cell cycle. Lack of zinc will lead to abnormal cell proliferation and then carcinogenesis. Through GSEA analysis, we found that a variety of cell cycle-related pathways were activated in high-risk group, which may be the reﬂection of zinc regulation of In this study, we analyzed the genomic changes of zinc metabolism-related genes in LUAD and constructed the ZMRGS. Frontiers in Immunology frontiersin.org 07 Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D E F A FIGURE 6 Immune-related molecular characteristics. (A) Chemokine. (B) Immunostimulator. (C) Immunoinhibitor. (D) MHC molecule. (E) Receptor. (F) Correlation circle of risk score and immune checkpoints. p < 0.05, p < 0.01, p < 0.001, **p < 0.0001. B A B D C D C F F E E FIGURE 6 Immune-related molecular characteristics. (A) Chemokine. (B) Immunostimulator. (C) Immunoinhibitor. (D) MHC molecule. (E) Receptor. (F) Correlation circle of risk score and immune checkpoints. p < 0.05, p < 0.01, p < 0.001, **p < 0.0001. activation on the whole. Abundant immune cell inﬁltration means that it is more likely to display a better response to immunotherapy (36). Subsequently, we predicted the response to immunotherapy through the TIDE algorithm. The results showed that, as we expected, the high-risk group had a higher TIDE score and was more prone to immune escape and the low-risk group showed excellent immunotherapy response (47.7% vs 27.3%). To verify the reliability of the results of TIDE, we used two datasets of anti-PD-1/PD-L1 immunotherapy for further analysis. The results, without exception, suggest that patients in the low-risk group had activation on the whole. Abundant immune cell inﬁltration means that it is more likely to display a better response to immunotherapy (36). Subsequently, we predicted the response to immunotherapy through the TIDE algorithm. 4 Discussion The results showed that, as we expected, the high-risk group had a higher TIDE score and was more prone to immune escape and the low-risk group showed excellent immunotherapy response (47.7% vs 27.3%). To verify the reliability of the results of TIDE, we used two datasets of anti-PD-1/PD-L1 immunotherapy for further analysis. The results, without exception, suggest that patients in the low-risk group had cell cycle disorder leading to poor prognosis of LUAD patients in our model. In addition, both non-speciﬁc and speciﬁc immune systems are affected by zinc, involving the proliferation, differentiation, and apoptosis of immune cells (35). Our research also found that there were a large number of immune pathways enriched and higher expression of immune molecules in the low- risk group, indicating that the low-risk group immune pathway was activated. The results of ssGSEA also conﬁrmed that various immune cells inﬁltrated in the low-risk group. These results indicate that the low-risk group was in the state of immune Frontiers in Immunology 08 frontiersin.org 10.3389/ﬁmmu.2023.1147528 Chang et al. B C D E F G H A FIGURE 7 Differences in TME and prediction of immunotherapy response. (A) The differences in the proportions of 28 immune cells between two ZMRGS groups by ssGSEA. Results of immune score (B), tumor purity (C), and stromal score (D) in two groups. (E) The results of TIDE score. (F) Response to immunotherapy from TIDE algorithm. Kaplan–Meier analysis by ZMRGS for patients in the IMvigor210 cohort (G) and Checkmate cohort (H). p < 0.05, p < 0.01, p < 0.001, **p < 0.0001. A A D D B B C C C B E E E F F H G G H H Differences in TME and prediction of immunotherapy response. (A) The differences in the proportions of 28 immune cells between two ZMRGS groups by ssGSEA. Results of immune score (B), tumor purity (C), and stromal score (D) in two groups. (E) The results of TIDE score. (F) Response to immunotherapy from TIDE algorithm. Kaplan–Meier analysis by ZMRGS for patients in the IMvigor210 cohort (G) and Checkmate cohort (H). p < 0.05, p < 0.01, p < 0.001, ***p < 0.0001. chemotherapy for the high-risk group may be an option to improve the prognosis of such patients. These results provided a reliable basis for our ZMRGS to be applied in clinical practice and provide different treatment guidance for different LUAD populations. Funding prediction of immunotherapy was only based on some machine algorithms. Finally, the potential role of these key zinc metabolism- related genes in lung cancer still needs further basic experimental exploration. This study was supported by the Guangdong Association Study of Thoracic Oncology (GASTO-202203). In conclusion, we have constructed a new and reliable zinc metabolism-related gene signature, which can effectively predict the prognosis of LUAD patients and distinguish the immune status of different LUAD patients. More importantly, it can distinguish and give LUAD patients with different immune states more appropriate treatment to improve their prognosis. Ethics statement All claims expressed in this article are solely those of the authors and do not necessarily represent those of their afﬁliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. This study was approved by the Ethics Committee of Sun Yat- sen University Cancer Center (YB2018-85). The patients/ participants provided their written informed consent to participate in this study. Written informed consent was obtained from the individual(s) for the publication of any potentially identiﬁable images or data included in this article. Acknowledgments We acknowledge TCGA and GEO databases for providing their platforms and contributors for uploading their meaningful datasets. Data availability statement The authors declare that the research was conducted in the absence of any commercial or ﬁnancial relationships that could be construed as a potential conﬂict of interest. The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary Material. 4 Discussion better prognosis when receiving immunotherapy. Although great progress has been made in immunotherapy, only 20% of patients with NSCLC respond to immune checkpoint inhibitors (ICIs) (37). Previous studies have conﬁrmed that ICIs combined with chemotherapy could improve the effectiveness of immunotherapy (38); hence, we also analyzed the sensitivity of chemotherapy drugs and found that the high-risk group was more sensitive to multiple chemotherapy drugs. Therefore, immunotherapy combined with There are some limitations in this study. First of all, the data we analyzed was from the public database, and further multi-center clinical trial veriﬁcation is still needed in the future. Secondly, due to the lack of transcriptome datasets of LUAD immunotherapy, the Frontiers in Immunology frontiersin.org 09 Chang et al. 10.3389/ﬁmmu.2023.1147528 B C D E F G H A FIGURE 8 Chemotherapy sensitivity analysis. (A) Cisplatin. (B) Docetaxel. (C) Doxorubicin. (D) Etoposide. (E) Paclitaxel. (F) Vinblastine. (G) Gemcitabine. (H) Vinorelbine. D C B A E F H F G G H E G FIGURE 8 Chemotherapy sensitivity analysis. (A) Cisplatin. (B) Docetaxel. (C) Doxorubicin. (D) Etoposide. (E) Paclitaxel. 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https://openalex.org/W2982338855	OpenAlex	Open Science	CC-By	2,019	Delineating the Regional Economic Geography of China by the Approach of Community Detection	Kang Wu	English	Spoken	11,166	18,203	Received: 17 September 2019; Accepted: 29 October 2019; Published: 31 October 2019 Received: 17 September 2019; Accepted: 29 October 2019; Published: 31 October 2019 Abstract: With the obvious regionalization trend in the new period of urbanization in China, the scientiﬁc delineation of functional regions (FRs) at diﬀerent scales has become a heated topic recently. Since the 20th century, western academia has formed a basic idea of metropolitan areas’ (MAs) delineation based on population density and commuting rate, for which the subjectivity of threshold setting is diﬃcult to overcome. In this study, community detection algorithms from the ﬁeld of network science are employed, namely the Louvain algorithm with adjustable resolutions and Combo with high-precision output, respectively. We take the nationwide car-hailing data set as an example to explore a bottom-up method for delineating regional economic geography at diﬀerent scales based on the interconnection strength between nodes. It was found that most of the prefecture-level cities in China have a dominant commuting region and two or three secondary commuting sub-regions, while regional central cities have extended their commuting hinterlands over jurisdictional boundaries, which is not common due to the larger initial administrative divisions and the comprehensive development niveau of cities. The feasibility and limitation of community detection partitioning algorithms in the application of regional science are veriﬁed. It is supposed to be widely used in regional delimitation supported by big data. Both of the two algorithms show a shortage of ignorance of spatial proximity. It is necessary to explore new algorithms that can adjust both accuracy and spatial distance as parameters. Keywords: commute pattern; community detection; regional delineation; big data; functional regions Sustainability 2019, 11, 6053; doi:10.3390/su11216053 sustainability sustainability sustainability sustainability Delineating the Regional Economic Geography of China by the Approach of Community Detection Kang Wu 1,* , Jingxian Tang 2 and Ying Long 3 1 Beijing Key Laboratory of Megaregions Sustainable Development Modelling and School of Urban Economics and Public Administration, Capital University of Economics and Business, Beijing 100070, China 2 School of Architecture, Tsinghua University, Beijing 100084, China; tjx14@mails.tsinghua.edu.cn 3 School of Architecture and Hang Lung Center for Real Estate, Tsinghua University, Beijing 100084, China; ylong@tsinghua.edu.cn 1 Beijing Key Laboratory of Megaregions Sustainable Development Modelling and School of Urban Economics and Public Administration, Capital University of Economics and Business, Beijing 100070, China 2 School of Architecture, Tsinghua University, Beijing 100084, China; tjx14@mails.tsinghua.edu.cn 3 School of Architecture and Hang Lung Center for Real Estate, Tsinghua University, Beijing 100084, China; ylong@tsinghua.edu.cn * C d k @ b d 1 Beijing Key Laboratory of Megaregions Sustainable Development Modelling and School of Urban Economics and Public Administration, Capital University of Economics and Business, Beijing 100070, China 2 School of Architecture, Tsinghua University, Beijing 100084, China; tjx14@mails.tsinghua.edu.cn 3 School of Architecture and Hang Lung Center for Real Estate, Tsinghua University, Beijing 100084, China; ylong@tsinghua.edu.cn y g g * Correspondence: wukang@cueb.edu.cn www.mdpi.com/journal/sustainability 1. Introduction The unexpected demographic change in the last 40 years has shown profound impacts on the morphology of urbanized areas around the world [1]. The agglomeration economic phenomenon has led to a speciﬁc manifestation of concentration in industries, population, and built-up areas around growth poles worldwide [2–5]. With the “intangible hand” of the market against the barriers formed by existing administrative jurisdictions, cities are integrated inextricably with the surrounding municipalities, which is now interpreted in terms of metropolitan area, metropolitan region, mega-region, megalopolis, or urban agglomeration in the context of highly dense and continuous urban in East Asia [6]. The concept of urban agglomeration (UA) evolved initially from Gottmann (1957) as the metropolis [7], with the following concept of conurbation proposed by Geddes (1968) [8]. Afterward, the discussion with respect to large-scale contiguous urbanized areas lasted for years in terms such as the mega-urban region [9], global city-region [10], polycentric city regions [11], mega-region [12–14] under diﬀerent social contexts. As an evolving concept, Fang argues a UA follows a spatiotemporal path from a single Sustainability 2019, 11, 6053; doi:10.3390/su11216053 www.mdpi.com/journal/sustainability 2 of 18 Sustainability 2019, 11, 6053 city to metropolitan areas (MAs), metropolitan area belts, large metropolitan area belts, and a megalopolis [6]. To summarize the common features described, urban agglomeration is recognized by one or more core cities with a population threshold and obvious functional dominance, surrounded contiguously by several big cities with which the core is closely associated. These highly integrated urbanized clusters build impacted connection through well-developed transportation and information and communications technology (ICT) infrastructure, which encourages daily commuting and ﬂows [1,6]. Compared to the matured UAs which entered the post-industrialization stage in the European and American continents, the phenomenon of UA has demonstrated more potential and diversity in Asian countries because population evolution has coincided with the process of globalization since the last decades of the 20th century [10]. According to the United Nations 2018 world population report, among the 33 mega-cities in the world, 20 are located in Asia [15]. UA is more likely to be the future form for the Asian urbanized settlement with universal signiﬁcance. To focus on the context of China, since the beginning of the reform and opening-up policy, central cities which had the privilege of the priority development policy have become the engine of the national economic growth by becoming involved in the global industry chain and global production networks [16]. 1. Introduction After 2000, the development of MAs and UAs has gained increasing importance with the shift in national, regional policies. The policy of urbanization was reversed for the ﬁrst time at the 16th Congress of the Communist Party in 2002, promoting the coordinated development of large, medium, and small cities. Soon after, in 2007, it encouraged the nation to “take full advantage of the leading role of mega-cities and large metropolitan areas and urban agglomerations, fostering new economic growth poles.” Most recently in November 2019, the “Opinions on Establishing a More Eﬀective Regional Coordination and Development Mechanism” clearly stated that a new model of regional development led by central cities and UAs, to promote long-term coordination and sustainability, and subsequently 19 national-level MAs/UAs planning strategies was approved [17]. However, despite the clariﬁcation of MAs/UAs in the national policy over time, the existing delineation has not reached consensus among geographers and planners. For instance, the empirical studies of various MAs/UAs’ delineation often have their own criteria and datasets, leaving a research gap for exploration until now. Given that the consistent standard for delineation is not indiﬀerent, the results are inevitably inﬂuenced by the measurement framework. To eliminate the ambiguity and inconsistency, this paper explores a new interdisciplinary empirical method employing the full-sample nationwide car-hailing commuting dataset as a proxy to delineate the regional economic geography in the context of China. Even though it is fundamental that the deﬁnition comes before measurement, it is an impossible task to completely portray the features of regional economic geography by one theory. As earlier geographers and economists determined the hierarchical structure within the geographical scope by the scale of the center [18,19], current regionalism shows more recognition of the theory of space of ﬂows and urban networks [20,21]. Based on such a functional connection hypothesis of geography, scholars have gradually reached a consensus which is to use connection as a proxy to characterize the functional interaction inside MAs/UAs for boundary delineation, such as information, enterprise headquarters and branches [22,23], transportation and daily commuting of the integrated labor market [11,24,25]. One of the main objectives of the study is to delineate the regional economic geography under current development conditions, to verify whether the hypothesis of economic activities spillover happens or not in the context of China. 2. Literature Review Regional delineation is a classic topic for geographers and other closely allied disciplines stretching back more than one century [28], which attempts to divide space into stable territorial regions relying on physical proximity, economic integration, and cultural similarity [25]. Due to the complex and highly heterogeneous distribution of features which characterize human geography [29], a regionalization scheme that combines both spatial laws and practical framework remains very much alive and unsolved. Instead of traditional contestable interpretive methods or semi-quantitative analysis based on the recognition of a hierarchical structure and bounded space, more recently, scholars have asserted the importance of ﬂows and networks to better understand the space [30]. Fueled by globalization, information and communications technology (ICTs), and high-speed transportation networks, the world has witnessed dramatic changes in the past three decades, which triggered a massive global time-space compression [31,32] and a new space forms emerged termed “the space of ﬂows” [20,33]. Compared to “the space of place”, the new space form shows a combined changing characteristic in both expanded geographical scope and intensiﬁed connections beyond the conﬁnes of boundary [34,35]. In the scope of urban and regional studies, Pﬂieger and Rozenblat (2010) raised their viewpoint of “The city as a network of networks” as a response to Berry’s “Cities as systems within systems of cities” proposed in 1964, which emphasized the relationality and connectivity in the new city space [36,37]. As a paradigm shift, the most prevailing concept and model are “world city networks” and “interlocking network model” proposed by GaWC (Globalization and World Cities Research Network), which oﬀer an empirical approach to measure numerous intercity connections and even delineate functional regions at diﬀerent scales [11,21]. Various data from corporate relations to transportation infrastructures were applied, while the methods have also evolved from distance-based gravity models to social network analysis [38–40]. However, the collection of these kinds of relational data is the “dirty little secret” for urban network research that might challenge the comparative analysis [41]. Exciting new advances in big data, especially the availability of large-scale open data from human interaction, social media, and mobility, have been experienced in the last ten years. Progress in information and communication technologies and the accumulation of massive data sets on human behavior now allow researchers to explore spatial interaction and complex patterns with unprecedented precision [42,43]. 1. Introduction Following the existing research, we try to explore the interdisciplinary application of community detection algorithms in the delimitation of functional regions through empirical evidence based on the accessible car-hailing commuting dataset provided by the main service provider in China. In addition to simply visualizing the origin–destination (O-D) ﬂow, this paper employs two network-partitioning algorithms with slightly diﬀerent accuracy and computation speed, namely Louvain and Combo [26,27]. These bottom-up computation approaches based on the relative relational strength of O-D pairs are expected to provide a relatively rigorous and consistent result of regional delineation covering nationwide. Additionally, it provides a new way of thinking for the delineation of boundaries in urban and regional studies and how to use the 3 of 18 Sustainability 2019, 11, 6053 emerging massive datasets to reduce the subjectivity of previous spatial sciences relying on experience. The remainder of the paper is as follows: We review the related literature from both scholarly studies and current policy practice aspects in Section 2. Section 3 introduces the data and methods of this research. Section 4 presents the main results from both commute landscape and community partitioning scope. Finally, discussions and conclusions are conducted in Sections 5 and 6, respectively. 2. Literature Review Such rich data from location-based services to social media services (i.e., mobile phone, digital footprint, smart card, and taxi trajectory data) deepens insights into human interactions in the era of “the space of ﬂows” by providing new data sources, inspiring new analytic methods and oﬀering new research perspectives [44–52], which otherwise would be impossible to understand quantitatively. Among these, some studies have focused on delineating geographical regions using both networks and community structure of human interaction, which has opened an entirely new window for those who are interested in such questions [25,43,53,54]. On the other hand, from planning and policy perspective, it is essential to deﬁne a coherent, bounded region to enable zoning, deﬁne electoral districts, and manage public services and municipal jurisdictions. One basic reason for identifying such units is that, in reality, the administrative boundary of a city/region cannot represent the actual size and impact of the labor force and economic activity happening there well. To capture its real population or economic functions, the concept of functional regions (FRs) or functional urban regions (FURs) are introduced and become more often used in urban policies. Among these, both metropolitan areas (MAs) and urban agglomerations (UAs) are typical regions whose geographical logic follows the underlying functional structure of these new patterns. Since the introduction of the concept of MAs and UAs, the bureaus of statistics and scholars of diﬀerent 4 of 18 Sustainability 2019, 11, 6053 countries have come up with a large set of methods to delineate their boundaries. The mainstream of these methods can be summarized as a two-step approach—ﬁrst, identify the core spatial unit, and then identify other spatial units, denoted as the hinterland, which have a signiﬁcant connection with the core. Nonetheless, although the basic idea of these methods resembles the speciﬁc data and techniques used, the identiﬁcation of the core or measurement of the strength of connections diﬀer in various ways. The U.S. Census Bureau’s delineation method can be summarized as a population-based core threshold, plus a bidirectional commuting threshold [55]. In Europe, the European Spatial Planning Observation Network (ESPON) endeavored to establish a uniform standard for the delineation of metropolitan areas, but as the data availability varies greatly among the countries, a unique standard is yet to be fully accomplished. 2. Literature Review While the density-based method for core identiﬁcation is still in its conception [56], the Economic Co-operation and Development (OECD) also uses population density to deﬁne the cores. However, for the same reason as the diﬀerent nature of administrative divisions—“unevenly sized and highly heterogeneous within and between countries” [57], it is diﬃcult to deﬁne a uniform deﬁnition for all the OECD members. In China, The National Bureau of Statistics of China has not published a national standard for the delineation of MAs and UAs. However, from the 1990s, Chinese scholars began focusing on similar topics, including deﬁning urban economic regions through to urban agglomerations [58–61]. The methods of Chinese scholars also followed the core–hinterland scheme. For the deﬁnition of the core there have been disputes, but almost all scholars used the statutory urban districts as the core. However, due to the lack of commuting data on a nationwide base, they used a variety of other measures of connection for the deﬁnition of the hinterland, including bus ridership, industrial economic indicators, and also road network [62]. Although the core–hinterland scheme is an eﬃcient and census-friendly way to delineate metropolitan areas, scholars have also criticized the assumptions in this two-step delineation method. Both the ex-ante selection of urban cores and the use of commuting as the single measure of connection were both challenged [4], not to mention that there is no accessible survey data on commuters in most Chinese cities. To avoid falling into the mud of the threshold discussion, this paper borrows the method of community detection based on modularity calculation, identifying FRs considering the relative strength of the connections, and a nationwide car-hailing commuting dataset is applied as a proxy to represent the commute landscape in China to delineate the functional regions (FRs) for the ﬁrst time. We also want to demonstrate the “natural” patterns of community clustering compare to the oﬃcial MAs/UAs through the lens of visual interpretations and network analysis. We hope our explorations provide a more empirically approach to detecting and deﬁning MAs/UAs and show the utility of this evaluation to gain a deep understanding of the regional economic geography of China. 3.2. Method 3 2 1 Id ifi i i h M h d f C i D i 3.2.1. Identiﬁcation using the Method of Community Detection 3.2.1. Identification using the Method of Community Detection The methods employed in this research are two borrowed algorithms of community detection, namely Louvain and Combo, from the field of network science. The calculations are mutually verified to increase the robustness and quasi-determination of the research. Community detection has its theoretical root in graph theory [63], focusing on the cluster partitioning and structure features of the various complex systems [64]. The increasing accessibility of huge amounts of internet-based data sets has increasingly promoted its prosperity and application in other fields, such as biology, social networks, technological networks, and spatial networks [65–67]. Visualizing the complexity of communities within a network has been recognized as crucial for scholars to understand its general features [65]. The methods employed in this research are two borrowed algorithms of community detection, namely Louvain and Combo, from the ﬁeld of network science. The calculations are mutually veriﬁed to increase the robustness and quasi-determination of the research. Community detection has its theoretical root in graph theory [63], focusing on the cluster partitioning and structure features of the various complex systems [64]. The increasing accessibility of huge amounts of internet-based data sets has increasingly promoted its prosperity and application in other ﬁelds, such as biology, social networks, technological networks, and spatial networks [65–67]. Visualizing the complexity of communities within a network has been recognized as crucial for scholars to understand its general features [65]. features [65]. A network or graph can be represented as a set of vertices, joined in pairs by edges [68]. Most of the community structure within a network follows a common rule that vertices show higher concentrations in specific groups while lower concentration between groups [65]. Communities, or modules, are groups of vertices sharing common properties and/or playing similar roles within the graph [69]. It has a very high similarity with the spatial relation demonstrated in the spatial science, that the urban core is always closely connected with a certain hinterland, while the link between two cities or two regions is loose and blurred. The in-between suburbs possibly have ambiguous connections on both sides. If we concentrate specifically on the commute flow, in reality, functional regions (FRs) could be considered as a network structure composed of commuter flows with the origin, destination, and directed segments forming the vertices and edges. 3.1. Car-hailing Commuting Dataset We used the accessible car-hailing commuting dataset provided by the most popular service provider in China to represent the commute landscape. Given that car-hailing has become one of the most convenient commute ways for people of a certain income level, the high-resolution dataset (to the scale of Jiedao/township) is supposed to provide the ﬁrst chance for us to portray commute connections clearly at a ﬁne scale. Since we focus on commuting connections inside those regions, we speciﬁcally considered the time period setting. What we employed was a car-hailing dataset of three consecutive workdays during the morning peak (7:00 to 9:30), which covered commute records of taxi-hailing service, tailored taxi, fast ride, and ride-sharing service. The four service forms almost represent diﬀerent income groups. Due to the coverage of diﬀerent incomes and popular use of this car-hailing service, we consider this data one of the best available commute datasets in current China. This dataset also divides the whole country into 53,979 transport statistics units (TSUs) representing towns. If we cluster the towns into larger units, we could get 338 prefecture-level city units and 2598 5 of 18 hina. Sustainability 2019, 11, 6053 car-hailing service, w county-level units, which are tested for regional delineation. The total number of traveling records during the commute period was 7,213,371, which accounted for 16.5% of all-day car-hailing travels (more than 41 million records in total). We connected all the O-D points of TSUs by ArcGIS and calculated the number of travel times in each distance interval. About 98.6% of the commuter ﬂow was less than 30 km, while 99.7% was within 80 km (7,194,509 in total, as shown in Figure 1). Considering the tolerance of commuters to time and price in reality, according to Nelson and Rae [25], the study believes that 80 km is a threshold to ﬁgure out whether the commute ﬂow has practical signiﬁcance or not. The methods used and results generated in the following parts are based on it. towns. If we cluster the towns into larger units, we could get 338 prefecture-level city units and 2598 county-level units, which are tested for regional delineation. The total number of traveling records during the commute period was 7,213,371, which accounted for 16.5% of all-day car-hailing travels (more than 41 million records in total). 3.1. Car-hailing Commuting Dataset We connected all the O-D points of TSUs by ArcGIS and calculated the number of travel times in each distance interval. About 98.6% of the commuter flow was less than 30 km, while 99.7% was within 80 km (7,194,509 in total, as shown in Figure 1.). Considering the tolerance of commuters to time and price in reality, according to Nelson and Rae [25], the study believes that 80 km is a threshold to figure out whether the commute flow has practical significance or not. The methods used and results generated in the following parts are based on it. Figure 1. Percentage of commuting flow volumes of different distance intervals. Figure 1. Percentage of commuting ﬂow volumes of diﬀerent distance intervals. Figure 1. Percentage of commuting flow volumes of different distance intervals. Figure 1. Percentage of commuting ﬂow volumes of diﬀerent distance intervals. 3.2.2. Algorithms Used for Regional Delineation Generally, algorithms of community partitions are valued by their computational precision within a reasonable time. The past 20 years have witnessed great progress in the development of well-performed algorithms. Famous algorithms in application include the simple greedy agglomerative optimization by Newman [68], the faster Clauset–Newman–Moore heuristic [70], and recently the Louvain method proposed by the Université catholique de Louvain, which provides an approach to ﬁnd high modularity partitions of networks with large amounts of nodes in a short time and demonstrates the hierarchical structure clearly [71]. For each node i, the algorithm considers the neighbors j of i and evaluates the gain of modularity that would take place by removing i from its community and by placing it in the community of j. The node i is then placed in the community for which this gain is maximum (in case of a tie, we use a breaking rule), but only if this gain is positive. If no positive gain is possible, i stays in its original community. This process is applied repeatedly and sequentially for all nodes until no further improvement can be achieved, and the ﬁrst phase is then complete. Its accuracy is veriﬁed as excellent in comparison and the algorithm has been applied widely until now [72–74]. In comparison, the recently emerged Combo method is an optimized algorithm with higher accuracy at the price of longer computational time proposed by Sobolevsky and Campari in 2014 [27]. The vast majority of search strategies take one of the following steps to evolve starting partitions: merging two communities, splitting a community into two, or moving nodes between two distinct communities. Combo involves all three possibilities, after selecting an initial partition made of a single community, the following steps are iterated as long as any gain in terms of the objective function score can be obtained: (1) For each source community, the best possible redistribution of every source nodes into each destination community (either existing or new) is calculated; this also allows for the possibility that the source community entirely merges with the destination; (2) the best merger, split, or recombination is performed. The modularity score of Combo is the best in the performance test, together with diﬀerent algorithms, including Louvain. Furthermore, it also shows similarity to Infomap when optimizing the description code length [27], indicating much better overall quality in practice [25]. 3.2.2. Algorithms Used for Regional Delineation Since our dataset provides us the ﬁne-scale computational resources to explore its mathematical properties, we employ both Louvain and Combo for the calculation to increase the robustness of the results and test their availability in the application of spatial science. For the calculation by Louvain, we transferred the original ﬁle into Comma-Separated Values ﬁle and kept the necessary value for analysis in Gephi (Source, Target, Type, Id, Label, Weight); while the realization of Combo was via the open-source code package developed and shared by the MIT’s Senseable City Lab (http://senseable.mit.edu/community_detection/). We had a total of 53,979 nodes, using an assembly computer with a memory of 16 GB and ubuntu16.04 × 64, the calculating of Combo cost 1015.34 s in total. We used Python to number the O-D nodes in order, converting origin and destination to “vertices” format and listing the segments as “arcs”. The results calculated by Louvain and Combo include the overall modularity and module group number that each vertice belongs to. We then used ArcGIS and QGIS to match and visualize the data. It should be pointed out that both algorithms have a shortcoming in the application of spatial science; that is, the spatial distance cannot be considered as the weight factor. Instead, the weight used in calculating is only the frequency of the same O-D arcs. 3.2. Method 3 2 1 Id ifi i i h M h d f C i D i 3.2.1. Identiﬁcation using the Method of Community Detection The “vertices” with higher A network or graph can be represented as a set of vertices, joined in pairs by edges [68]. Most of the community structure within a network follows a common rule that vertices show higher concentrations in speciﬁc groups while lower concentration between groups [65]. Communities, or modules, are groups of vertices sharing common properties and/or playing similar roles within the graph [69]. It has a very high similarity with the spatial relation demonstrated in the spatial science, that the urban core is always closely connected with a certain hinterland, while the link between two cities or two regions is loose and blurred. The in-between suburbs possibly have ambiguous connections on both sides. If we concentrate speciﬁcally on the commute ﬂow, in reality, functional regions (FRs) could be considered as a network structure composed of commuter ﬂows with the origin, destination, and directed segments forming the vertices and edges. The “vertices” with higher centrality sharing a large number of edges are supposed to be signiﬁcant as the core area/job market inside an FRs, while vertices lying at the boundaries with limited edges connected to others may play the role as a hinterland or 6 of 18 Sustainability 2019, 11, 6053 represent places of residence. Considering the attribute of the commute ﬂow network, we believe that the algorithms of community detection are suitable and feasible for regional delineation in China. 4.1. The Overall Visualized Commute Landscape 4.1. The Overall Visualized Commute Landscape We ﬁrst conducted a visualization of the car-hailing commuting dataset to get a rough depiction of the overall distribution of China’s current commute patterns. Visualizing all trips less than 80 km nationwide (Figure 2), the commute landscape conﬁrmed the existence of some larger metropolitan areas and urban agglomerations that mainly concentrate on the coastal area previously under priority development policy, which accounted for 55.1%. In addition, Western and Central China is witnessing the formation of Chengdu, Xi’an, Chongqing, Wuhan, Changsha, Zhengzhou metropolitan areas. However, the overall commuting ﬂow proportion of the central, western and northeastern region was still low, constituting 19.7%, 17.3%, and 7.8%, respectively, and the metropolitanization reﬂected by commuting was centered around several major cities, reﬂecting the characteristics of China’s urbanization in the previous stage. Most of the large metropolitan areas show characteristics with a unique cluster, for example, Zhengzhou, Hefei, Wuhan, and Changsha metropolitan Areas, while Qingdao, Dalian, and Xiamen perform as multiple clusters inside the boundary. We first conducted a visualization of the car-hailing commuting dataset to get a rough depiction of the overall distribution of China's current commute patterns. Visualizing all trips less than 80 km nationwide (Figure 2.), the commute landscape confirmed the existence of some larger metropolitan areas and urban agglomerations that mainly concentrate on the coastal area previously under priority development policy, which accounted for 55.1%. In addition, Western and Central China is witnessing the formation of Chengdu, Xi’an, Chongqing, Wuhan, Changsha, Zhengzhou metropolitan areas. However, the overall commuting flow proportion of the central, western and northeastern region was still low, constituting 19.7%, 17.3%, and 7.8%, respectively, and the metropolitanization reflected by commuting was centered around several major cities, reflecting the characteristics of China's urbanization in the previous stage. Most of the large metropolitan areas show characteristics with a unique cluster, for example, Zhengzhou, Hefei, Wuhan, and Changsha metropolitan Areas, while Qingdao, Dalian, and Xiamen perform as multiple clusters inside the boundary. Figure 2. The car-hailing commute landscape of 80 km or less in China. Figure 2. The car-hailing commute landscape of 80 km or less in China. Figure 2. The car-hailing commute landscape of 80 km or less in China. Figure 2. The car-hailing commute landscape of 80 km or less in China. The commuting volume is verified to be positively correlated with economic patterns and population density. 4. Results In the following section, we present the results by both algorithms after a brief visualization analysis of the general commuting landscape of the whole country and four main policy urban agglomerations. Afterward, we present the results of the Louvain and Combo approaches to detect 7 of 18 f 18 Sustainability 2019, 11, 6053 Sustainability 2019, 11, x FO the functional regions/clusters based on the car-hailing commuting ﬂows inter and intra regions. With diﬀerent resolutions, the algorithmic method was veriﬁed to be available to demonstrate potential FRs at various scales, namely (larger) metropolitan areas or even urban agglomerations. If the rationality of the proxy data is further improved, the eﬀectiveness of the method will inevitably be further enhanced. agglomerations. Afterward, we present the results of the Louvain and Combo approaches to detect the functional regions/clusters based on the car-hailing commuting flows inter and intra regions. With different resolutions, the algorithmic method was verified to be available to demonstrate potential FRs at various scales, namely (larger) metropolitan areas or even urban agglomerations. If the rationality of the proxy data is further improved, the effectiveness of the method will inevitably be further enhanced. 4.1. The Overall Visualized Commute Landscape 4.1. The Overall Visualized Commute Landscape Refining the scale on the four main urban agglomerations namely Yangtze River Delta (22.5%), Pearl River Delta (9.9%), Beijing–Tianjin–Hebei Region (15.7%), and Chengdu- Chongqing Region (12.2%), we could form a basic understanding of the central cities, linkage strengths of nodes, and comprehensive development level in each UA. The former two UAs have already presented as polycentric city regions in process, whereas the Beijing–Tianjin–Hebei Region has three core cities, namely Beijing, Tianjin, and Shijiazhuang, while the former two are closely connected and account for more than 80% of all commuter flows in this region. Chengdu and Chongqing still develop with less interconnection between each other due to the overall size of the economy (Figure 3.). The commuting volume is veriﬁed to be positively correlated with economic patterns and population density. Reﬁning the scale on the four main urban agglomerations namely Yangtze River Delta (22.5%), Pearl River Delta (9.9%), Beijing–Tianjin–Hebei Region (15.7%), and Chengdu-Chongqing Region (12.2%), we could form a basic understanding of the central cities, linkage strengths of nodes, and comprehensive development level in each UA. The former two UAs have already presented as polycentric city regions in process, whereas the Beijing–Tianjin–Hebei Region has three core cities, namely Beijing, Tianjin, and Shijiazhuang, while the former two are closely connected and account for more than 80% of all commuter ﬂows in this region. Chengdu and Chongqing still develop with less interconnection between each other due to the overall size of the economy (Figure 3). 8 of 18 Sustainability 2019, 11, 6053 Figure 3. The commute landscape of the four main urban agglomerations (UAs) in China Figure 3. The commute landscape of the four main urban agglomerations (UAs) in China. Figure 3. The commute landscape of the four main urban agglomerations (UAs) in China Figure 3. The commute landscape of the four main urban agglomerations (UAs) in China. The above visual approach provides us a simple but clear image with spatial patterns and structure. We demonstrated it here as a basis for the subsequent algorithmic results. The following parts will separately present the result by Louvain and Combo, to identify the availability and pros and cons of regional delineation methods by employing community detection algorithms. 4 2 R i li i b L i The above visual approach provides us a simple but clear image with spatial patterns and structure. We demonstrated it here as a basis for the subsequent algorithmic results. 4.2. Regionalization by Louvain The computation process 4.2. Regionalization by Louvain The computation process of the Louvain algorithm is completed quickly in Gephi software. With different resolutions as modularity parameters (we tried 1, 3, 5, 7, 11, 20, 50, and 100), we finally got the modularity results with a different number of communities detected and modularity resolution as follow (Table 1.): T bl 1 R l b h L i l i h The computation process of the Louvain algorithm is completed quickly in Gephi software. With diﬀerent resolutions as modularity parameters (we tried 1, 3, 5, 7, 11, 20, 50, and 100), we ﬁnally got the modularity results with a diﬀerent number of communities detected and modularity resolution as follow (Table 1): y g Parameters/Resolution 0.1 1 3 5 7 11 20 50 100 Number of Communities 762 614 569 556 548 542 531 520 511 Modularity with Resolution 0.074 0.958 2.954 4.951 6.949 10.944 19.94 49.943 99.926 Modularity 0.882 0.958 0.956 0.956 0.955 0.952 0.944 0.943 0.941 We also visualize the community partition results in, each TSU (township) with different colors indicates it is partitioned into a different community. To understand the difference more clearly, we focus on the Beijing Tianjin Hebei Region Yangtze Ri er Delta and Pearl Ri er Delta as the main Table 1. Result by the Louvain algorithm. Parameters/Resolution 0.1 1 3 5 7 11 20 50 100 Number of Communities 762 614 569 556 548 542 531 520 511 Modularity with Resolution 0.074 0.958 2.954 4.951 6.949 10.944 19.94 49.943 99.926 Modularity 0.882 0.958 0.956 0.956 0.955 0.952 0.944 0.943 0.941 y g Parameters/Resolution 0.1 1 3 5 7 11 Table 1. Result by the Louvain algorithm. focus on the Beijing Tianjin Hebei Region, Yangtze River Delta, and Pearl River Delta as the main observing regions. With the highest accuracy of Louvain, only the northern three counties of Langfang city and the towns of Baoding on the boundary of Beijing are classified to the main Beijing region, which means that the hinterland of Beijing is just slightly crossing administrative boundaries while the other prefecture-level cities have their own unique commuting hinterland within the jurisdiction. On the one hand, China’s prefecture-level administrative divisions are large in scale, as we mentioned before. In addition, it also reflects the barriers formed by the division of the administrative powers. 4.1. The Overall Visualized Commute Landscape 4.1. The Overall Visualized Commute Landscape The following parts will separately present the result by Louvain and Combo, to identify the availability and pros and cons of regional delineation methods by employing community detection algorithms. 4.2. Regionalization by Louvain The computation process 4.2. Regionalization by Louvain Even in the Yangtze River Delta, the most market-oriented region in China, We also visualize the community partition results in, each TSU (township) with diﬀerent colors indicates it is partitioned into a diﬀerent community. To understand the diﬀerence more clearly, we focus on the Beijing–Tianjin–Hebei Region, Yangtze River Delta, and Pearl River Delta as the main observing regions. With the highest accuracy of Louvain, only the northern three counties of Langfang city and the towns of Baoding on the boundary of Beijing are classiﬁed to the main Beijing region, which means that the hinterland of Beijing is just slightly crossing administrative boundaries while the other prefecture-level cities have their own unique commuting hinterland within the jurisdiction. On the one hand, China’s prefecture-level administrative divisions are large in scale, as we mentioned 9 of 18 Sustainability 2019, 11, 6053 before. In addition, it also reﬂects the barriers formed by the division of the administrative powers. Even in the Yangtze River Delta, the most market-oriented region in China, the cross-border region is seldom detected when the parameter setting is strict. What we detected by the Louvain algorithm were basically the same as the jurisdiction boundaries, except that the FR of Hangzhou covered the eastern part of Huzhou city and the border area together with Shaoxing, while the rest of Shaoxing city integrated into the FR of Ningbo, and prefecture-level cities in central Jiangsu Province had a relatively small area of jurisdiction but closer links in between. In comparison, the integration level of daily commuting in the Pearl River Delta was close to the borderless urban area; Guangzhou, Foshan, Dongguan, and Shenzhen and border area of Zhaoqing, Qingyuan have already formed into one cluster (Figure 4). Sustainability 2019, 11, x FOR PEER REVIEW 9 of 18 the cross-border region is seldom detected when the parameter setting is strict. What we detected by the Louvain algorithm were basically the same as the jurisdiction boundaries, except that the FR of Hangzhou covered the eastern part of Huzhou city and the border area together with Shaoxing, while the rest of Shaoxing city integrated into the FR of Ningbo, and prefecture-level cities in central Jiangsu Province had a relatively small area of jurisdiction but closer links in between. 4.2. Regionalization by Louvain The computation process 4.2. Regionalization by Louvain In comparison, the integration level of daily commuting in the Pearl River Delta was close to the borderless urban area; Guangzhou, Foshan, Dongguan, and Shenzhen and border area of Zhaoqing, Qingyuan have already f d i t l t (Fi 4) ( g ) Figure 4. Computational result of functional regions (FRs) by Louvain when Parameters/Resolution =1 Figure 4. Computational result of functional regions (FRs) by Louvain when Parameters/Resolution =1. Figure 4. Computational result of functional regions (FRs) by Louvain when Parameters/Resolution Figure 4. Computational result of functional regions (FRs) by Louvain when Parameters/Resolution =1. Sustainability 2019, 11, 6053 10 of 18 Comparing the FR pattern of diﬀerent resolution intervals, four or more FRs joined into one, as shown in Figure 5. The FR computational result by the largest resolution interval provides us a clue of the objective commuting integrated urban agglomerations in China. Within the Beijing–Tianjin–Hebei Region, Baoding aﬃliated towns and cities along the Beijing–Guangzhou expressway, the whole jurisdiction of Langfang city and Zhangjiakou city and Tianjin aﬃliated Jixian were integrated into the Beijing FR, while Tangshan and the rest of Tianjin and Cangzhou city form another FR surrounding the Bohai Bay. The Yangtze River Delta comprised four FR sections with Shanghai, Hangzhou, Ningbo, and Nanjing as the core, while the commuter FR at the largest scale of the Pearl River Delta exceeded the deﬁned boundary written in the policy (Figure 5), implying an even larger economic hinterland in the future. Sustainability 2019, 11, x FOR PEER REVIEW 10 of 18 of the objective commuting integrated urban agglomerations in China. Within the Beijing–Tianjin– Hebei Region, Baoding affiliated towns and cities along the Beijing–Guangzhou expressway, the whole jurisdiction of Langfang city and Zhangjiakou city and Tianjin affiliated Jixian were integrated into the Beijing FR, while Tangshan and the rest of Tianjin and Cangzhou city form another FR surrounding the Bohai Bay. The Yangtze River Delta comprised four FR sections with Shanghai, Hangzhou, Ningbo, and Nanjing as the core, while the commuter FR at the largest scale of the Pearl River Delta exceeded the defined boundary written in the policy (Figure 5), implying an even larger economic hinterland in the future. Figure 5. Computational result of FRs by Louvain when Parameters/Resolution =100. Figure 5. Computational result of FRs by Louvain when Parameters/Resolution =100. Figure 5. Computational result of FRs by Louvain when Parameters/Resolution =100. Figure 5. 4.2. Regionalization by Louvain The computation process 4.2. Regionalization by Louvain Computational result of FRs by Louvain when Parameters/Resolution =100. 11 of 18 Sustainability 2019, 11, 6053 11 of 18 4.3. Regionalization by Combo 4.3. Regionalization by Combo Sustainability 2019, 11, x FOR PEER REVIEW 12 of 18 may have more than five FRs; the influencing spheres of regional cities with functional importance have extended across the administrative boundaries, for example, Wuhan, Xi’an, Zhengzhou, and so on (Figure 7). From the results of the measurement, the Combo algorithm with a high-precision parameter setting can be applied to the city-region scale to assist in identifying spatial sub-clusters within a prefecture-level city, reducing the subjectivity relying on planners’ experience. Figure 7. Refined scope of FR result by Combo with the highest accuracy. 4.4. Comparing Larger FRs Tested by Louvain and Combo with Policy-Defined Regions Figure 7. Reﬁned scope of FR result by Combo with the highest accuracy. 4.4. Comparing Larger FRs Tested by Louvain and Combo with Policy-Deﬁned Regions Figure 7. Refined scope of FR result by Combo with the highest accuracy. Figure 7. Reﬁned scope of FR result by Combo with the highest accuracy. g p y g y 4 4 Comparing Larger FRs Tested by Louvain and Combo with Policy-Defined Regions 4.4. Comparing Larger FRs Tested by Louvain and Combo with Policy-Deﬁned Regions p g g y y f g In addition to the above-mentioned results detected with town-scale car-hailing data, we also tried to increase "resolution" by merging town-scale data into county-level and city-level before applying the community detection algorithms. However, due to the limitations of the data itself that the total number of car-hailing data across the prefecture-level city was insignificant, the result was not ideal. By contrast, two different partitioning patterns have been achieved on the county scale by Louvain and Combo, respectively ( Figure 8; Figure 9). The results under the standard resolution of Louvain provide a larger regional division that characterizes the relative strength between cities, most of which have exceeded the recent policy-defined metropolitan circles that are, in fact, the 1- hour accessible area of the regional central cities [75]. The Combo algorithm still generated an image of the metropolitan area, located in or slightly beyond the administrative boundary of the prefecture- level city, while the scale was much smaller and inconsistent compared to the questionable et o olita i le i the oli y In addition to the above-mentioned results detected with town-scale car-hailing data, we also tried to increase "resolution" by merging town-scale data into county-level and city-level before applying the community detection algorithms. 4.3. Regionalization by Combo 4.3. Regionalization by Combo The result obtained by the Combo algorithm (Figure 6) shows an obviously higher accuracy than that of the Louvain algorithm at the cost of approximately 17 minutes of computing time. We kept the precision parameter as the original “inﬁnity” and got 183 regions detected as the pattern below with a modularity of 0.9643. It should be noted that the Combo algorithm removed points that cannot be counted or not suitably counted into a community, so there was a signiﬁcant diﬀerence in the number of the recognized communities from Louvain. The highest precision results were basically consistent with the perception of the metropolitan area deﬁned by the daily commuting rate. The result obtained by the Combo algorithm (Figure 6) shows an obviously higher accuracy than that of the Louvain algorithm at the cost of approximately 17 minutes of computing time. We kept the precision parameter as the original “infinity” and got 183 regions detected as the pattern below with a modularity of 0.9643. It should be noted that the Combo algorithm removed points that cannot be counted or not suitably counted into a community, so there was a significant difference in the number of the recognized communities from Louvain. The highest precision results were basically consistent with the perception of the metropolitan area defined by the daily commuting rate. Figure 6. FR computational result by Combo with the highest accuracy. Figure 6. FR computational result by Combo with the highest accuracy. Figure 6. FR computational result by Combo with the highest accuracy. Figure 6. FR computational result by Combo with the highest accuracy. 12 of 18 Sustainability 2019, 11, 6053 Most of the prefecture-level cities had a unique larger urban commuting area, with one or two smaller sub-clusters. Cities with complex topography or a larger jurisdiction, such as Chongqing city, may have more than ﬁve FRs; the inﬂuencing spheres of regional cities with functional importance have extended across the administrative boundaries, for example, Wuhan, Xi’an, Zhengzhou, and so on (Figure 7). From the results of the measurement, the Combo algorithm with a high-precision parameter setting can be applied to the city-region scale to assist in identifying spatial sub-clusters within a prefecture-level city, reducing the subjectivity relying on planners’ experience. 4.3. Regionalization by Combo 4.3. Regionalization by Combo However, due to the limitations of the data itself that the total number of car-hailing data across the prefecture-level city was insigniﬁcant, the result was not ideal. By contrast, two diﬀerent partitioning patterns have been achieved on the county scale by Louvain and Combo, respectively (Figures 8 and 9). The results under the standard resolution of Louvain provide a larger regional division that characterizes the relative strength between cities, most of which have exceeded the recent policy-deﬁned metropolitan circles that are, in fact, the 1-hour accessible area of the regional central cities [75]. The Combo algorithm still generated an image of the metropolitan area, located in or slightly beyond the administrative boundary of the prefecture-level city, while the scale was much smaller and inconsistent compared to the questionable metropolitan circle in the policy. 13 of 18 of 18 of 18 Sustainability 2019, 11, 6053 Sustainability 2019 11 x FO Sustainability 2019, 11, x FO Figure 8. Larger FRs tested by Louvain and county-level clustered data. Figure 8. Larger FRs tested by Louvain and county-level clustered data. Figure 8. Larger FRs tested by Louvain and county-level clustered data. Figure 8. Larger FRs tested by Louvain and county-level clustered data. Figure 8. Larger FRs tested by Louvain and county-level clustered data. Figure 8. Larger FRs tested by Louvain and county level clustered data. Fi u e 9 L FR t t d b C b d t l l l t d d t Figure 9. Larger FRs tested by Combo and county-level clustered data. Figure 9. Larger FRs tested by Combo and county-level clustered data. Figure 9. Larger FRs tested by Combo and county-level clustered data. Figure 9. Larger FRs tested by Combo and county-level clustered data. 5. Discussion 5. Discussion 5. Discussion Using the new emerging car-hailing data set, this study explored the application scenarios of algorithm-based community detection methods in urban and regional studies. And the results indicate this exploration can indeed identify functional regions of different scales according to the relative connection strength between nodes employing massive data set. It could outline the boundary directly, avoiding the cumbersome calculating process of the traditional methods by defining the urbanized center and its periphery by population threshold first and then recognize the commuting hinterland with the accurate commuting percentage towards the functional center, what Using the new emerging car-hailing data set, this study explored the application scenarios of algorithm-based community detection methods in urban and regional studies. And the results indicate this exploration can indeed identify functional regions of different scales according to the relative connection strength between nodes employing massive data set. It could outline the boundary directly, avoiding the cumbersome calculating process of the traditional methods by defining the urbanized center and its periphery by population threshold first and then recognize the commuting hinterland with the accurate commuting percentage towards the functional center, what Using the new emerging car-hailing data set, this study explored the application scenarios of algorithm-based community detection methods in urban and regional studies. And the results indicate this exploration can indeed identify functional regions of diﬀerent scales according to the relative connection strength between nodes employing massive data set. It could outline the boundary directly, avoiding the cumbersome calculating process of the traditional methods by deﬁning the urbanized center and its periphery by population threshold ﬁrst and then recognize the commuting hinterland with the accurate commuting percentage towards the functional center, what is more, it is also supposed to avoid the subjectivity of pre-setting centers of a region. Compared to the previous research conducted by Gu (1991) [58], Zhou and Zhang (2003) [60], and Ning (2011) [59], our research can identify potential 14 of 18 Sustainability 2019, 11, 6053 14 of 18 city regions, metropolitan areas, and urban agglomerations by diﬀerent parameter settings, which addresses the needs of regional boundaries delineation for planning and regional policy pre-research at diﬀerent scales well. However, from another perspective, it is crucial to realize that the applicable scenarios of the two algorithms are both closely related to the setting of their parameters. The accuracy and credibility of the results depend on the researcher’s personal understanding of regions. 5. Discussion 5. Discussion It is foreseeable that if we employ better proxy data set, the detected boundaries by our approach will be much closer to reality. With continuous time-series commuting data set, we could also observe the dynamic process of regional spatial evolution, which is obviously maneuverable in the era of massive increasingly accessible data. Using the computational methods borrowed from network science, a clearer spatial deﬁnition and delineation is going to be achieved, which prompts scholars to reach a wider consensus on the issue of the regional boundary, thus easing the disputes and deepening the further discussion referring to the same spatial context. There are still two major limitations in the research so far. First, the proxy of data remains insuﬃcient. According to the estimation, the proportion of car-hailing commute in the overall commuting ﬂow is not dominant, so we cannot say that the regional delineation is completely solved. Taking more comprehensive full-sample proxy data, such as rail transit data, cell phone signaling data, telephony data, email, mail, or other data sets, that can characterize daily economic connection as agents is expected to have more satisfying results in the follow-up research. Second, the deﬁciencies of the algorithm for community detection and the ambiguity of parameter settings have not been overcome. If we return to the algorithm generation logic, it fails to take the spatial distance into consideration, which means that the highly valued “spatial proximity” in regional science is almost ignored. During the calculation process, all nodes were treated as indistinguishable points in contrast to that in reality. Cities and towns that are close to each other integrate more in terms of economy, culture, and social interaction, therefore, it is ﬂawed to neglect the attribute of spatial proximity. In addition, as mentioned above, the accuracy of the application of the community detection algorithm is rooted in its parameter setting. Expect for more straightforward and scientiﬁc presentation, the regional delineation results under diﬀerent parameter settings with those measured needed to further veriﬁed and corrected by other reliable methods. Otherwise, regional delineation still partially depends unavoidably on the subjective knowledge and empirical judgment of the researcher. 6. Conclusions This paper employed the nationwide commuting proxy dataset for the ﬁrst time to carefully identify the current functional regions at diﬀerent scales. By the computational results of the two algorithms, we can conclude that in China, most prefecture-level cities have a major FR together with two to three smaller sub-clusters. The phenomenon of commuting beyond the boundaries of municipal administrative scope is relatively rare except for central cities that undertake functions as regional economic centers, the FRs of which are commonly extended along the traﬃc corridors to the border areas of nearby prefecture-level cities, such as Zhengzhou, Wuhan, Xi’an, Chengdu, thus demanding the coordination of regional governance. The internal commuting pattern of the three major urban agglomerations is diﬀerent from what is deﬁned by the policy. The FRs of Beijing only reach out slightly to a few towns in the east and south. The relationship between Beijing and Tianjin is not like the pattern suggested by the previous visualization. Cities with close functional linkages with Tianjin locate around the Bohai Bay, while the hinterland of Beijing is mainly towns within 100 km of Beijing, especially the eastward. As one of the world’s largest urbanized continuous zone, daily commuting in the Pearl River Delta is closely compacted into integrated regions, in particular, the Foshan–Guangzhou–Dongguan urbanized area. For the two algorithms veriﬁed in this paper, Combo shows better accuracy than Louvain. Furthermore, Combo is tested to be able to cover and observe a larger scale despite the longer computational time. On the other hand, the study explored a refreshing method of regional delineation by employing interdisciplinary knowledge in the context of China for the ﬁrst time. We are not in the succession 15 of 18 15 of 18 Sustainability 2019, 11, 6053 of traditional delineation methods proposed by previous studies since the end of the 20th century, which follows the steps of identifying centers and hinterlands by population and then calculating commuting rate as the threshold. Instead, we focused more on the linkage intensity between city nodes within an FR. In addition, the nationwide commuting data set provided us a chance to inspect the bias of current regional policy. We have also depicted the overall structure of the densely urbanized area and FRs of central cities that we have ignored for a long time. 6. Conclusions If the accuracy of the data set is further increased and then overlapped with other economic, geographic information, this approach is expected to provide scientiﬁc support for regional policy development at the provincial level. Author Contributions: Conceptualization, K.W.; Methodology, K.W. and J.T.; Software, J.T. and K.W.; Resources, Y.L. and K.W.; Writing—original draft preparation, J.T. and K.W.; Writing—Review and Editing, K.W.; Visualization, J.T. and K.W.; Supervision, K.W.; Funding acquisition, K.W. Funding: This research was funded by the Natural Science Foundation of China, grant number 41671161/41401178/71733001, a research project of Institute for China Sustainable Urbanization, Tsinghua University, grant number K-17009-1, and the Youth Talent Support Program in University of Beijing, grant number CIT&TCD201804097. Acknowledgments: The authors appreciate Beijing Xiaoju Technology Co, Ltd. for providing car-hailing dataset. Conﬂicts of Interest: The authors declare no conﬂict of interest. Acknowledgments: The authors appreciate Beijing Xiaoju Technology Co, Ltd. for providing car-hailing dataset. 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https://openalex.org/W4391137989	OpenAlex	Open Science	CC-By	null	Copyright Protection for AI-Generated Works: Exploring Originality and Ownership in a Digital Landscape	H. Abdul Gaffar	English	Spoken	17,723	27,547	© The Author(s), 2024. Published by Cambridge University Press on behalf of the Asian Society of International Law. This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/ licenses/by/4.0/), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited. Hafiz GAFFAR and Saleh ALBARASHDI Hafiz GAFFAR and Saleh ALBARASHDI Sultan Qaboos University, Muscat, Oman Corresponding author: Hafiz GAFFAR; E-mail: h.ibrahim@squ.edu.om Sultan Qaboos University, Muscat, Oman Corresponding author: Hafiz GAFFAR; E-mail: h.ibrahim@squ.edu.om (Received 3 July 2023; revised 19 October 2023; accepted 2 November 2023) Asian Journal of International Law (2024), 1–24 doi:10.1017/S2044251323000735 Asian Journal of International Law (2024), 1–24 doi:10.1017/S2044251323000735 * Hafiz GAFFAR is an Assistant Professor in Civil Law, College of Law, at Sultan Qaboos University, Muscat, Oman. Saleh ALBARASHDI is an Associate Professor in Commercial Law, College of Law at Sultan Qaboos University. 1 A.M. TURING, “Lecture to the London Mathematical Society”, in D.C. INCE, ed., Mechanical Intelligence: Collected Works of A.M. Turing (North Holland, 1992). 2 Tianxiang HE, “The sentimental fools and the fictitious authors: Rethinking the copyright issues of AI-generated contents in China”, (2019) 27.2 Asia Pacific Law Review 218. * Hafiz GAFFAR is an Assistant Professor in Civil Law, College of Law, at Sultan Qaboos University, Muscat, Oman. Saleh ALBARASHDI is an Associate Professor in Commercial Law, College of Law at Sultan Qaboos University. * Hafiz GAFFAR is an Assistant Professor in Civil Law, College of Law, at Sultan Qaboos University, Muscat, Oman. Saleh ALBARASHDI is an Associate Professor in Commercial Law, College of Law at Sultan Qaboos University. 1 A.M. TURING, “Lecture to the London Mathematical Society”, in D.C. INCE, ed., Mechanical Intelligence: Collected Works of A.M. Turing (North Holland, 1992). 2 Tianxiang HE, “The sentimental fools and the fictitious authors: Rethinking the copyright issues of AI-generated contents in China”, (2019) 27.2 Asia Pacific Law Review 218. © The Author(s), 2024. Published by Cambridge University Press on behalf of the Asian Society of International Law. This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/ licenses/by/4.0/), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited. 2 Tianxiang HE, “The sentimental fools and the fictitious authors: Rethinking the copyright issues of AI-generated contents in China”, (2019) 27.2 Asia Pacific Law Review 218. y 1 A.M. TURING, “Lecture to the London Mathematical Society”, in D.C. INCE, ed., Mechanical Intelligence: Collected Works of A.M. Turing (North Holland, 1992). 2 Tianxiang HE, “The sentimental fools and the fictitious authors: Rethinking the copyright issues of https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Abstract This research explores AI-generated originality’s impact on copyright regulations. It meticulously examines legal frameworks such as the Berne Convention, EU Copyright Law, and national legisla- tion. Rigorously analyzing cases, including Infopaq International A/S v Danske Dagblades Forening and Levola Hengelo BV v Smilde Foods BV, illuminates evolving originality and human involvement in AI creativity. The study also contemplates global perspectives, drawing from esteemed organiza- tions such as the World Intellectual Property Organization and the European Court of Justice and exploring diverse approaches adopted by individual nations. The paper emphasizes the imperative need for legislative updates to address the challenges and opportunities of AI-generated works. It highlights the pivotal role of international collaboration and public awareness in shaping copyright policies for the AI-driven creativity era. It also offers insights and recommendations for policymakers and researchers navigating this complex terrain. Keywords: Copyright law; artificial intelligence; authorship; ownership; human author; non-human author; originality; copyrightability; Berne Convention; AI-generated works. Artificial intelligence (AI), as defined by A.M. Turing, refers to the development of com- puter systems that can perform tasks requiring human-like intelligence.1 In recent years, there has been a surge of interest in the legal issues surrounding AI-generated content, particularly about copyright. The rapid advancement of AI technology has led to ques- tions regarding the legal status and protection of AI-generated works.2 AI has emerged as a transformative force, revolutionizing various aspects of human life, including creating and producing artistic and literary works. However, existing copyright legislation in most * Hafiz GAFFAR is an Assistant Professor in Civil Law, College of Law, at Sultan Qaboos University, Muscat, Oman. Saleh ALBARASHDI is an Associate Professor in Commercial Law, College of Law at Sultan Qaboos University. 1 A.M. TURING, “Lecture to the London Mathematical Society”, in D.C. INCE, ed., Mechanical Intelligence: Collected Works of A.M. Turing (North Holland, 1992). 2 Tianxiang HE, “The sentimental fools and the fictitious authors: Rethinking the copyright issues of AI-generated contents in China”, (2019) 27.2 Asia Pacific Law Review 218. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 2 2 countries cannot protect AI-generated works, which include creative works produced with AI technology and even computer-generated works. Abstract These involve creative content gen- erated exclusively by computer algorithms or programs and fail to address the complex issues surrounding AI-generated works.3 Undoubtedly, human authors form the foundation of copyright, and, as such, the law protects them in recognition of their originality and creativity.4 Hence, the author’s identification is important as it is a prerequisite for “copyrightability”. Granting authors copyright entails bestowing on them two distinct sets of rights. The first set enables authors to reap the financial rewards of their creative endeavors, such as selling copy- rights. The second set encompasses moral and enduring rights, including the right of paternity, which allows authors to be recognized by name, use pseudonyms, and remain anonymous, and the right to integrity, which empowers authors to refuse modifications that could harm their reputation.5 Consequently, the foundation of copyright and intellectual property law rests on the notion of human “authorship”. However, this established legislative policy has been sig- nificantly disrupted by the emergence of AI and its growing involvement as human assis- tants in creating diverse forms of works, including literary and artistic creations. As a result, the determination of who should be recognized as the author of such works has become a highly contentious matter, not only for policymakers but also for judicial sys- tems.6 Given the significant impact of AI-generated works and their substantial role in the creative process, it is now essential and unavoidable for legislative policymakers to estab- lish a clear legal framework that defines the precise legal relationship between computers and their operators or programmers. By doing so, courts will be empowered to address the authorship of computer-generated works and allocate ownership rights accordingly.7 Notably, certain countries, such as the UK, Hong Kong, Ireland, New Zealand, and India, have taken significant steps and initiatives to address the issue of AI-generated works. These jurisdictions have taken proactive measures by amending their statutes to accom- modate the copyrightability of works that involve computer-generated elements and human intervention. By doing so, they have surpassed the international legal community in providing legal clarity in this area.8 Nevertheless, these laws do not comprehensively address those scenarios where a computer creates a work with minimal or indirect human involvement, potentially leaving a lacuna in the legal framework. 3 Yurii BURYLO, “AI-generated works and copyright protection”, (2022) 3 Entrepreneurship, Economy and Law 7. 4 The legal frameworks adopted with regard to attributing copyright to AI-generated works would strike at the core of the social purpose for which the copyright system existed. See WIPO Secretariat, “Revised Issued Paper on Intellectual Property Policy and Artificial Intelligence”, WIPO/IP/AI/2/GE/20/1 REV (11 June 2022), para. 23. 5 Sam RICKETSON and Jane C. GINSBURG, International Copyright and Neighboring Rights: The Berne Convention and Beyond, 2nd ed. (New York: Oxford University Press, 2006) 87, at 586. 6 Jane C. GINSBURG, “The Concept of Authorship in Comparative Copyright Law”, (2003) 52 DePaul L. Rev. 1,063, at 1,066. 7 Pamela SAMUELSON, “Allocating Ownership Rights in Computer-Generated Works”, (1986) 47 U. Pitt. L. Rev. 1,185, at 1,189–90. 8 See s 9(3) of the Copyright, Designs, and Patents Act (CDPA) of 1988; s 5 of New Zealand Copyright Act (1994); s 11(3) of Copyright Ordinance (CO) (Cap. 528); s 21( f ) of Copyright and Related Rights Act (2000). 9 Daniel BÖTTICHER, “Copyright Protection for Artificial Intelligence Output”, (2019), Master Thesis, University of St. Gallen) at 33–5. 8 See s 9(3) of the Copyright, Designs, and Patents Act (CDPA) of 1988; s 5 of New Zealand Copyright Act (199 1(3) of Copyright Ordinance (CO) (Cap. 528); s 21( f ) of Copyright and Related Rights Act (2000). Yurii BURYLO, AI-generated works and copyright protection , (2022) 3 Entrepreneurship, Economy and Law 7. 4 The legal frameworks adopted with regard to attributing copyright to AI-generated works would strike at the core of the social purpose for which the copyright system existed. See WIPO Secretariat, “Revised Issued Paper on Intellectual Property Policy and Artificial Intelligence”, WIPO/IP/AI/2/GE/20/1 REV (11 June 2022), para. 23. y y 6 Jane C. GINSBURG, “The Concept of Authorship in Comparative Copyright Law”, (2003) 52 DePaul L. Rev. 1,063, at 1,066. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press y g y g g 9 Daniel BÖTTICHER, “Copyright Protection for Artificial Intelligence Output”, (2019), Master Thesis, University of St. Gallen) at 33–5. 3 Yurii BURYLO, “AI-generated works and copyright protection”, (2022) 3 Entrepreneurship, Economy a 4 h l l f k d d h d b h d k ld Abstract By contrast, most jur- isdictions worldwide still struggle to grapple with the complex issue of authorship and remain silent on this contentious matter.9 This article aims to delve into the intricate realm of AI authorship and explore how the emergence of smart technologies may disrupt the existing copyright framework. Specifically, it focuses on determining the authorship of works created by AI. A 3 Yurii BURYLO, “AI-generated works and copyright protection”, (2022) 3 Entrepreneurship, Economy and Law 7. 4 The legal frameworks adopted with regard to attributing copyright to AI-generated works would strike at the core of the social purpose for which the copyright system existed. See WIPO Secretariat, “Revised Issued Paper on Intellectual Property Policy and Artificial Intelligence”, WIPO/IP/AI/2/GE/20/1 REV (11 June 2022), para. 23. 6 Jane C. GINSBURG, “The Concept of Authorship in Comparative Copyright Law”, (2003) 52 DePaul L. Rev. 1,063, at 1,066. 7 Pamela SAMUELSON, “Allocating Ownership Rights in Computer-Generated Works”, (1986) 47 U. Pitt. L. Rev. 1,185, at 1,189–90. 7 Pamela SAMUELSON, “Allocating Ownership Rights in Computer-Generated Works”, (1986) 47 U. Pitt. L. Rev. 1,185, at 1,189–90. 8 See s 9(3) of the Copyright, Designs, and Patents Act (CDPA) of 1988; s 5 of New Zealand Copyright Act (1994); s 11(3) of Copyright Ordinance (CO) (Cap. 528); s 21( f ) of Copyright and Related Rights Act (2000). 8 See s 9(3) of the Copyright, Designs, and Patents Act (CDPA) of 1988; s 5 of New Zealand Copyright Act (1994); s 11(3) of Copyright Ordinance (CO) (Cap. 528); s 21( f ) of Copyright and Related Rights Act (2000). 9 Daniel BÖTTICHER, “Copyright Protection for Artificial Intelligence Output”, (2019), Master Thesis, University of St. Gallen) at 33–5. 9 Daniel BÖTTICHER, “Copyright Protection for Artificial Intelligence Output”, (2019), Master Thesis, University of St. Gallen) at 33–5. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Asian Journal of International Law 3 comprehensive analysis of the prerequisites of authorship within the established legal system is necessary to address this question. Through an analytical approach, this article carefully examines the landscape of copyright law and judicial practices in various coun- tries, including the US, the UK, and China. Additionally, it considers the copyright frame- work at the EU level and the judicial practices of the Court of Justice of the European Union (CJEU). The selection of these jurisdictions was based on several key considerations. Abstract The jurisdictions were chosen based on their significant impact on developing and inter- preting copyright law. These jurisdictions have well-established copyright systems and have previously dealt with emerging technologies, making them relevant for our analysis. Additionally, jurisdictions were selected to represent different legal systems or approaches to copyright law, enabling a comparative analysis of how they address the issue of authorship in AI-generated works. Furthermore, consideration was given to jur- isdictions where AI-generated works had been subject to legal disputes or where ongoing debates on authorship existed, ensuring the practical relevance of the analysis. The avail- ability of research resources, including scholarly articles, legal opinions, and case law for the selected jurisdictions, was also taken into account to facilitate a comprehensive exam- ination of the legal principles surrounding authorship. By exploring these jurisdictions, this article seeks to provide insights into the current landscape of copyright law relating to AI-generated works. It investigates the role of judicial interpretations in establishing principles to address the deadlock surrounding authorship in AI-generated works. This article aims to contribute to the field in multiple ways by exploring different jurisdictions under civil and common law legal systems. First, it strives to provide a comprehensive understanding of the attribution of authorship in cases where the production involves human intervention or is autonomously generated. Such an understanding is valuable to judges, lawyers, and legal scholars working in copyright law. Second, the critical ana- lysis conducted can highlight potential challenges that may arise in judicial practice. This information can then be utilized to enhance judicial practices and ensure justice’s effect- ive and efficient delivery. However, while there has been considerable discourse and research on the legal issues surrounding AI-generated works and copyright, there appears to be a research gap in examining the specific criteria and implications of originality in AI-generated works within different legal jurisdictions. Existing literature primarily focuses on the general intersection of AI and copyright law, with limited attention given to the nuanced require- ments of originality for AI-generated content. Furthermore, there is a lack of comprehen- sive analysis and comparison of the approaches taken by different jurisdictions in addressing the copyright protection of AI-generated works. Our research seeks to address this gap by critically examining the concept of originality in AI-generated works and exploring the divergent legal approaches across various jurisdictions, specifically focusing on the implications for copyright law. Abstract As we navigate these complex issues, it is important to emphasize that this paper represents an ongoing dialogue in a rapidly evolving field. The conclusions and insights drawn will contribute to the broader discussions surrounding AI and copyright law, serv- ing as a foundation for further exploration and the development of legal frameworks that can effectively address the challenges posed by AI-generated authorship. 10 Pamela SAMUELSON, “AI Authorship?”, (2020) 63.7 Communications of the ACM 20, at 22. 11 National Commission on New Technological Uses of Copyrighted Works (CONTU), “Final Report on the National Commission on New Technological Uses of Copyrighted Works”, (1981) 3 Computer L. J. 53. 12 Ibid., at 54. 13 Annemarie BRIDY, “Coding Creativity: Copyright and the Artificially Intelligent Author”, (2012) 5 Stan. Tech. L. Rev. 1, 5, and 7. 14 U.S. Congress, Office of Technology Assessment, “Intellectual Property Rights in an Age of Electronics and Information”, OTA-CIT-302 (Washington, D.C.: U.S. Government Printing Office, 1986) (II). 15 Ibid. 16 Ibid. I. The Orthodox Concept of “Authorship” and the Premature Consideration of AI’s Capability to Create Literary And Artistic Works In the realm of literary and artistic works, the concept of authorship has traditionally relied upon the significance of human creativity. Consequently, human authors are https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 4 granted certain exclusive rights over their original creations. These rights encompass the ability to publish their work, reap the financial rewards, transfer ownership of copyrights, and safeguard their reputation by preventing any actions that could harm it. The Berne Convention stands as a cornerstone in the realm of copyright protection, offering a com- pelling example of the principles underlying the authorship concept. This international treaty ensures that literary and artistic works receive global protection, extending its ben- efits to authors and their successors in title. Doing so empowers authors, granting them the authority to assert their rights and safeguard their creative expressions. The Berne Convention further promotes transparency by encouraging the inclusion of clear indica- tions of the author’s name or pseudonym, effectively eliminating any ambiguity surround- ing the author’s identity. Consequently, the Berne Convention assumes a pivotal role in advocating for the safeguarding, recognition, and preservation of the contributions made by authors in the realm of literature and art. Together with other international treaties, the Berne Convention lays the foundational framework for comprehending authorship and copyright on a global scale. The unresolved questions surrounding the author’s identification and copyright attri- bution have given rise to significant attention from intellectual property (IP) experts. Works created with the aid of software have been a subject of discussion within the field of intellectual property for several decades, particularly regarding the role of AI or computers as tools. Despite the passage of time, these fundamental questions remain unanswered, creating a need for further examination and clarification.10 In 1978, the US established the Commission on New Technological Uses of Copyrighted Works (CONTU) to address challenges arising from technological advancements.11 CONTU proposed key amendments to copyright law, recognizing computer programs as eligible for copyright protection and extending copyright provisions to cover all uses of copyrighted programs in computer-related activities. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 17 Alison FLOOD, “‘A box of light’: AI inspired by British verse attempts to write poetry”, 20 March 2021, The Guardian, online: The Guardian https://www.theguardian.com/books/2021/mar/20/a-box-of-light-ai-inspired- by-british-verse-attempts-to-write-poetry. 18 Jacob BROGAN, “An A.I. Competed for a Literary Prize, but Humans Still Did the Real Work”, 25 March 2016, Slate, online: Slate https://slate.com/technology/2016/03/a-i-written-novel-competes-for-japanese-literary- award-but-humans-are-doing-the-work.html. 19 Melissa AVDEEFF, “Artificial Intelligence & Popular Music: SKYGGE, Flow Machines, and the Audio Uncanny Valley”, (2019) 8(4), Arts 130.1, at 11. 20 Samuel FISHWICK, “Robot rock: How AI singstars use machine learning to write harmonies”, 1 March 2018, The Standard, online: The Standard https://www.standard.co.uk/tech/jukedeck-maching-learning-ai-startup- music-a3779296.html. I. The Orthodox Concept of “Authorship” and the Premature Consideration of AI’s Capability to Create Literary And Artistic Works Additionally, it emphasized the importance of ensuring that the rightful possessors of copies of computer programs have the freedom to use and adapt those copies within copyright law bounds.12 p p py g In 1986, the Office of Technology Assessment (OTA) studied emerging developments of the technology’s impact on intellectual property.13 OTA focused on the federal copyright system’s efficacy in addressing emerging technologies,14 concluding that technological advancements significantly impacted the intellectual property system. These advance- ments necessitated ongoing adaptation of laws to keep pace with the evolving techno- logical landscape.15 The emergence of AI algorithms has revolutionized the impact of new technologies. These algorithms have become ubiquitous, influencing various aspects of life extending beyond mere predictive capabilities for the future. In essence, the aspirations of the past have become a reality. The transformative power of AI algorithms has reshaped our understanding of technological progress and its profound influence on society.16 Drawing insights from the challenges posed by utilizing new technologies is crucial. One notable illustration is an intriguing poem created through the capabilities of AI https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 5 Asian Journal of International Law algorithms. In this instance, British scientists developed a program that assimilates verses from over one hundred contemporary British poets, enabling it to learn and emulate the distinctive poetry writing styles of acclaimed poets like Simon Armitage and Alice Oswald. The machine was able to generate its own poems. A representative example of poetry gen- erated by AI is presented below:17 and soon I am staring out again, begin to practice my words, expecting my word will come. it will not. the wind is calling. my friend is near; I hear his breath. his breath is not the air. he touches me again with his hands and tells me I am growing old, he says, far old. we travel across an empty field in my heart. there is nothing in the dark, I think, but he. I close my eyes and try to remember what I was he says it was an important and interesting day, because I put in his hands one night the box of light that had been a tree. In 2016, another captivating illustration of machine-generated content emerged with the publication of a novel by a Japanese AI program titled The Day a Computer Writes a Novel. I. The Orthodox Concept of “Authorship” and the Premature Consideration of AI’s Capability to Create Literary And Artistic Works Remarkably, this novel garnered significant recognition and was even shortlisted for the prestigious “Star New First Prize” award.18 The creation of SKYGGE’s Hello World serves as a noteworthy example that highlights the convergence of AI and human collab- oration. This groundbreaking achievement marked the first-ever fully AI-generated song.19 The company Jukedeck utilized machine learning to train its system on the prin- ciples of music theory, enabling it to compose various genres of music, including jazz, blues, and rock.20 This evidence strongly suggests that AI-generated creations are no longer confined to the realm of dreams. Consequently, it is imperative to consider the implications of AI-generated works within the framework of copyright law. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 21 Sam RICKETSON, “The 1992 Horace S. Manges Lecture - People or Machines: The Bern Convention and the Changing Concept of Authorship”, (1991) 16 Colum. VLA J. L., Arts 1, 37. 22 Ibid. 23 P. Bernt HUGENHOLTZ and João Pedro QUINTAIS, “Copyright and Artificial Creation: Does EU Copyright Law Protect AI-Assisted Output?” (2021) 52.9 IIC-International Review of Intellectual Property and Competition Law 1,190, at 1,195–6. 24 Berne Convention for the Protection of Literary and Artistic Works, 4 May 1896, 828 U.N.T.S. 221 (entered into force 29 January 1970) [Berne Convention]. 25 Paris Act relating to the Berne Convention for the Protection of Literary and Artistic Works, 9 September 1886, 3 U.N.T.S. 1,161 (entered into force 15 December 1972). 26 Caroline ANFRAY et al., “Reflection paper on copyright, patient-reported outcome instruments and their translations”, (2018) 16 Health and Quality of Life Outcomes 1. 27 Paul GOLDSTEIN, International Copyright: Principles, Law, and Practice, (New York, USA: Oxford University Press, 2001). II. AI-Works and the Riddles of Authorship The evolution of the copyright law system has witnessed significant transformations, from the advent of the printing press to the emergence of AI, which stands out as a pivotal milestone in its history. Amid this backdrop, the remarkable ability of AI applications to create literary and artistic works has triggered a global disruption within copyright sys- tems. The reverberations of this disruption are palpable in the ongoing discourse sur- rounding the intricate matter of authorship about the works generated by AI. Professor Sam Ricketson’s foresight led him to delve into the intriguing question of the “true https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 6 6 author” behind works created by AI: was it the result of human ingenuity or the creative prowess of machines? This prescient inquiry resonates deeply with the current conun- drum surrounding authorship in AI-generated works21. In his analysis, Professor Ricketson made an observation about the Berne Convention. He argued that the Convention did not explicitly define the author as a human being, as it was considered a self-evident truth among its members. This implicit assumption, while seemingly innocuous, raises thought-provoking questions regarding the evolving nature of author- ship in the face of AI-generated works.22 In recent years, the prevalence and effectiveness of AI applications in creating literary and artistic works has reached unprecedented levels. This exponential growth has given rise to a host of complex issues at the intersection of humans and machines, raising profound policy concerns within the realm of copyright systems. Fundamental questions have emerged, such as the preservation of traditional copyright principles, the potential impact of AI on human-related copyrights, and the determination of authorship when a non-human entity is the creator or composer of a work. 21 Sam RICKETSON, “The 1992 Horace S. Manges Lecture - People or Machines: The Bern Convention and the Changing Concept of Authorship”, (1991) 16 Colum. VLA J. L., Arts 1, 37. 22 Ibid A. Should Copyright be Granted to AI-Generated Literary and Artistic Works, or Should Human Involvement be Mandatory? For a literary or artistic work to qualify for legal protection, it must exhibit a certain level of creativity attributable to the author. The concept of authorship is intricately linked to the requirement of originality and personal creative input in creating the work. This cri- terion serves as a fundamental principle in determining the eligibility of a work for copy- right protection.23 The Berne Convention, a foundational international treaty for copyright protection, ensures that literary and artistic works are granted protection glo- bally, extending its benefits to authors and their successors in title.24 By empowering authors, the Berne Convention grants them the authority to enforce their rights and safe- guard their creative expressions. It encourages the inclusion of clear indications of the author’s name or pseudonym, thereby eliminating any ambiguity surrounding the author’s identity.25 As a result, the Berne Convention plays a pivotal role in promoting the protection, recognition, and preservation of the contributions made by authors in the realm of literature and art. Although the Berne Convention does not explicitly define the term “author”, it is widely understood that the absence of such a definition is not a limitation to human beings alone. The context and indications within the convention suggest that the term encompasses both human authors26 and legal entities.27 The focus of the Berne Convention is to ensure that the rights of creators, whether individuals or organizations, https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 7 Asian Journal of International Law are protected and that their creative works receive the benefits of copyright protection. Thus, while the Convention does not explicitly define the term “author”, it encompasses a broad understanding that includes both human and legal entity creators.28 The sources of copyright law in the European Union (EU) include international treaties, Union legisla- tion, and national law. The rights granted to creators of original works vary across EU Member States, influenced by their national law and legal tradition.29 The concept of the “author” in EU copyright law can vary depending on the legal tradition followed in each Member State. In the continental European model, inspired by the French “droit d’au- teur”, authorship encompasses moral and economic rights. Moral rights, such as the right to identification and integrity, are inalienable and cannot be transferred or waived. Economic rights, on the other hand, can be conveyed. A. Should Copyright be Granted to AI-Generated Literary and Artistic Works, or Should Human Involvement be Mandatory? Economic rights take precedence over moral rights in common law systems, which include the UK, Ireland, Malta, and Cyprus. The right to claim authorship and object to modifications are recognized as moral rights in British and Irish law, but these are granted rights that can be waived.30 Therefore, determin- ing who qualifies as an “author” in EU copyright law depends on the specific legal frame- work and the extent to which moral and economic rights are considered.31 According to Chinese copyright laws, copyright protects the producers of original artistic and literary works. The creator of a work automatically enjoys copyright protection from the moment the work is created. Copyright grants the author or creator exclusive rights, including the right to reproduce, distribute, exhibit, perform, broadcast, adapt, translate, and exploit the work for economic gain. The concept of authorship in copyright law refers to the individual who created the original work; the author is the rightful owner of the copyright and holds the moral and economic rights associated with the work. Therefore, the authorship of artistic and literary works is protected under copyright law, ensuring that the original creators are recognized and have control over their works.32 US copyright law protects authorship of literary and artistic works under Title 17 of the US Code. This protection encompasses original works of authorship fixed in any tangible medium of expression, including literary, musical, dramatic, pictorial, graphic, sculptural, motion pic- tures, sound recordings, and architectural works.33 Based on the laws discussed earlier, par- ticularly in the context of the Berne Convention, EU copyright law, and copyright laws in China, and the US, it becomes evident that the concept of authorship is primarily centred around human involvement. These laws emphasize the recognition and protection of the rights of human creators as the original authors of literary and artistic works. 28 The wording of the above text strongly suggests that “author” and “authorship” for the purposes of the Convention refer to the individual who created the work. As a result, it seems acceptable to say that copyright protection is exclusively granted to man-made creative works. See Ginsburg, supra note 6 at 1069. Convention refer to the individual who created the work. As a result, it seems acceptable to say that copyright protection is exclusively granted to man-made creative works. See Ginsburg, supra note 6 at 1069. 29 Comparative Law Library Unit, European Parliamentary Research Service, “Copyright Law in the EU: Salient Features of Copyright Law Across the EU Member States” (2018), European Parliament online: European Parliament https://www.europarl.europa.eu/RegData/etudes/STUD/2018/625126/EPRS_STU(2018)625126_EN.pdf. 30 See sections 77 and 80 of the Copyright, Designs and Patents Act (CDPA) 1988, Legislation.gov.uk online: Legislation.gov.uk https://www.legislation.gov.uk/ukpga/1988/48/contents. 31 Ibid. 32 China IPR SME Helpdesk, “Copyright Protection in China: A Guide for European SMEs”, China IPR Helpdesk online: European Commission https://intellectual-property-helpdesk.ec.europa.eu/system/files/ 2020-10/EN_Copyright_guide_Aug_2010.pdf. 33 “Copyright Law of the United States, and Related Laws Contained in Title 17 of the United States Code”, U.S. Copyright Office (2022) online: U.S. Copyright Office https://www.copyright.gov/title17/title17.pdf. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 29 Comparative Law Library Unit, European Parliamentary Research Service, “Copyright Law in the EU: Salient Features of Copyright Law Across the EU Member States” (2018), European Parliament online: European Parliament https://www.europarl.europa.eu/RegData/etudes/STUD/2018/625126/EPRS_STU(2018)625126_EN.pdf. 32 China IPR SME Helpdesk, “Copyright Protection in China: A Guide for European SMEs”, China IPR Helpdesk online: European Commission https://intellectual-property-helpdesk.ec.europa.eu/system/files/ 2020-10/EN_Copyright_guide_Aug_2010.pdf. B. What is the Concept of an “Author” in Common Law Countries? In countries that have a common law system, such as the US, the UK, Ireland, Malta, and New Zealand, there are discussions and debates regarding the recognition of non-human 28 The wording of the above text strongly suggests that “author” and “authorship” for the purposes of the Convention refer to the individual who created the work. As a result, it seems acceptable to say that copyright protection is exclusively granted to man-made creative works. See Ginsburg, supra note 6 at 1069. 29 Comparative Law Library Unit, European Parliamentary Research Service, “Copyright Law in the EU: Salie atures of Copyright Law Across the EU Member States” (2018), European Parliament online: European Parliame tps://www europarl europa eu/RegData/etudes/STUD/2018/625126/EPRS STU(2018)625126 EN pdf 29 Comparative Law Library Unit, European Parliamentary Research Service, “Copyright Law in the EU: Salient Features of Copyright Law Across the EU Member States” (2018), European Parliament online: European Parliament https://www.europarl.europa.eu/RegData/etudes/STUD/2018/625126/EPRS_STU(2018)625126_EN.pdf. 30 See sections 77 and 80 of the Copyright, Designs and Patents Act (CDPA) 1988, Legislation.gov.uk onlin gislation.gov.uk https://www.legislation.gov.uk/ukpga/1988/48/contents. 30 See sections 77 and 80 of the Copyright, Designs and Patents Act (CDPA) 1988, Legislation.gov.uk online: Legislation.gov.uk https://www.legislation.gov.uk/ukpga/1988/48/contents. 31 30 See sections 77 and 80 of the Copyright, Designs and Patents Act (CDPA) 1988, Legislation.gov.uk online: Legislation.gov.uk https://www.legislation.gov.uk/ukpga/1988/48/contents. 31 Ibid. 32 China IPR SME Helpdesk, “Copyright Protection in China: A Guide for European SMEs”, China IPR Helpdesk online: European Commission https://intellectual-property-helpdesk.ec.europa.eu/system/files/ 2020-10/EN_Copyright_guide_Aug_2010.pdf. 32 China IPR SME Helpdesk, “Copyright Protection in China: A Guide for European SMEs”, China IPR Helpdesk online: European Commission https://intellectual-property-helpdesk.ec.europa.eu/system/files/ 2020-10/EN_Copyright_guide_Aug_2010.pdf. 33 “Copyright Law of the United States, and Related Laws Contained in Title 17 of the United States Code”, U.S. Copyright Office (2022) online: U.S. Copyright Office https://www.copyright.gov/title17/title17.pdf. 33 “Copyright Law of the United States, and Related Laws Contained in Title 17 of the United States Code”, U.S. Copyright Office (2022) online: U.S. Copyright Office https://www.copyright.gov/title17/title17.pdf. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 8 8 authors in copyright law. These countries follow a utilitarian theory that focuses on incentivizing innovation and providing public access to creative works for the benefit of society. This perspective places less emphasis on the personality or identity of the author, potentially opening the door for non-human authors. However, examining the case law in these countries, particularly in the US, clarifies the stance taken on this issue. B. What is the Concept of an “Author” in Common Law Countries? The US Supreme Court, in its interpretation of the Copyright Act, has consistently maintained that copyright protection is limited to works created by human authors. This judicial stance narrows the scope of copyright protection and restricts its application to creations that stem from human creativity. An illustrative example is Burrow-Giles Lithographic Co. v Sarony,34 where the court defined an author as the “originator” or “maker” to whom a work owes its origin. The court specifically referred to photographs as representatives of the original intellectual conceptions of an author. Similarly, in the case of Goldstein v California, the Supreme Court reaffirmed that an author, in the consti- tutional sense, refers to an individual who writes or composes an original work. The term “author” was interpreted as the “originator” to whom the work owes its origin. These legal precedents emphasize the importance of human involvement and creativity in the concept of authorship. They establish that the term “author” is closely tied to the act of creation, attributing the origin of a work to an individual who exercises intellectual effort and originality. The decisions highlight the requirement for a human originator, excluding non-human entities from being recognized as authors within the constitutional framework. The case of Feist Publications, Inc. v Rural Telephone Service Company, Inc.35 further clarified the requirement for copyright protection. The court concluded that a work must be ori- ginal to the author to be eligible for copyright. In other words, the work must demon- strate sufficient creativity and originality attributable to the author. This ruling reinforces the notion that copyright protection is contingent upon the presence of origin- ality, emphasizing the importance of individual creative expression. It establishes that mere factual information or data compilation without the requisite creative input does not meet the threshold for copyright protection. Therefore, to qualify for copyright, a work must exhibit a distinctive and independently created expression from the author. p y p The settled case law of the Supreme Court has consistently underscored the require- ment of human authorship, and the lower courts have reiterated this position. The lower courts have consistently rejected the possibility of non-human authors aligning themselves with established legal precedents. 34 Burrow-Giles Lithographic Co. v Sarony, 111 U.S. 53 (1884). 35 Feist Publications, Inc., v Rural Telephone Service Co., 499 U.S. 340 (1991). 36 Urantia Foundation v Maaherra, 114 F.3d 955, 964 (9th Cir. 1997). 37 Cetacean Community v Bush, 249 F. Supp. 2d 1206 (D. Haw. 2003). B. What is the Concept of an “Author” in Common Law Countries? In the case of Urantia Foundation v Maaherra,36 the Ninth Circuit Court of Appeals ruled that a book containing words pur- portedly “authored” by non-human spiritual beings can only be eligible for copyright pro- tection if there is evidence of human selection and arrangement of the content. The court emphasized the importance of human involvement in the creative process and required a significant level of human input for copyright protection to be granted. In the case of Cetacean Community v Bush,37 a ruling was issued in 2018 that addressed the question of legal standing for animals, specifically a monkey, in relation to copyright claims. The court concluded that the monkey had standing under Article III of the US Constitution, which pertains to the jurisdiction of federal courts. However, the court held that the mon- key, like all animals, lacked statutory standing under the federal Copyright Act. This case highlighted the distinction between constitutional standing and statutory standing in the context of copyright law. While the monkey was deemed to have constitutional standing https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Asian Journal of International Law 9 to bring a lawsuit, the court determined that the statutory framework of the Copyright Act did not extend copyright protection to non-human animals. Therefore, the monkey’s claim for copyright infringement was ultimately dismissed based on the lack of statutory standing. In the case of Kelley v Chicago Park District,38 the court held that a copyright pro- tection claim was invalid in the context of a living garden. The court reasoned that a liv- ing garden does not possess the necessary attributes of authorship and stability required to meet the criteria for copyright protection. Unlike fixed and tangible work, such as a written composition or a sculpture, a garden is a dynamic entity that is inherently culti- vated and subject to seasonal changes. Therefore, it was determined that the garden was not sufficiently composed or installed to be considered a permanent work deserving of copyright protection. This case exemplifies the limitations of copyright law in recognizing certain types of works that do not conform to traditional notions of authorship and sta- bility. While copyright law aims to protect original creative works, including literary, art- istic, and architectural works, the court’s decision in Kelley highlights the inherent challenges in extending copyright protection to living and evolving creations such as gar- dens. 38 Kelley v Chicago Park District, 635 F.3d 290, 304 (7th Cir. 2011). 39 Copyright Review Board, U.S. Copyright Office, “Re: Second Request for Reconsideration for Refusal to Register a Recent. Entrance to Paradise” (Correspondence ID 1-3ZPC6C3; SR # 1-7100387071), 14 February 2022. 40 Blake BRITTAIN, “Getty Images lawsuit says Stability AI misused photos to train AI”, 7 February 2023, Reuters, online: Reuters https://www.reuters.com/legal/getty-images-lawsuit-says-stability-ai-misused-photos- train-ai-2023-02-06/. 41 “Generative Artificial Intelligence and Copyright Law”, (29 September 2023) Congressional Research Service online: Congressional Research Service https://crsreports.congress.gov/product/pdf/LSB/LSB10922. 42 Sam TOBIN, “Getty asks London court to stop UK sales of Stability AI system”, 2 June 2023, Reuters, online: Reuters https://www.reuters.com/technology/getty-asks-london-court-stop-uk-sales-stability-ai-system-2023- 06-01/. 43 “Getty Images v Stability AI”, BakerHostetler online: Baker Law https://www.bakerlaw.com/getty-images-v- stability-ai/. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press C. The Dilemma of AI Authorship under Asian Law In the context of Asian copyright systems, the concept of attributing authorship to humans is evident. China, known for its significant advancements in AI and extensive use of AI programs, encountered the question of authorship in relation to AI-generated works in legal proceedings. One notable case is Shenzhen Tencent v Shanghai Yingxun,44 where the Nanshan District People’s Court had to determine whether AI-created works should be eligible for copyright protection. The court’s ruling acknowledged that works produced by AI applications such as Dream Writer deserve copyright protection. However, it emphasized that the individual asserting authorship must fulfil the general requirement of intellectual creativity under Chinese law to claim authorship. Another significant case involving Gao Yang and Youku emerged before the Beijing Court. The central question raised in this case pertained to the copyrightability of photos taken automatically by a camera attached to a hot-air balloon. The court was tasked with determining whether these AI-generated photos could be protected under Chinese copy- right law. In its ruling, the court concluded that the automatic photos captured by the plaintiff’s camera were indeed eligible for copyright protection in accordance with China’s copyright law. Consequently, the defendant’s unauthorized use of these photos was deemed to be an infringement of the plaintiff’s copyright.45 Undoubtedly, the analysis of this judgment aligns with the overall direction because it recognized that, despite the photos being taken automatically by the hot air balloon, the plaintiff was still granted protection due to their minimal contribution. Their act of attaching a sports camera to the hot-air balloon was considered a sufficient creative input to qualify for copyright protection. p Similarly, in India, copyright law attributes authorship for computer-generated liter- ary, dramatic, or artistic works to the person who undertakes the necessary arrangements for their creation. However, it is important to note that the human author is still consid- ered the driving force behind the scenes, even though AI may have played a significant role in the creative process.46 India’s 1957 Copyright Act47 does not explicitly define “author” for artistic and literary works. However, section 2(d) of the Act provides a com- prehensive understanding of the term. According to the Act, the “author” of a literary or dramatic work refers to the person who created the work, while for a musical work, it refers to the composer. 44 (2019) Yue 0305 Min Chu No. 14010. 45 (2017) Jing 73 Min Zhong No. 797. 46 Andres GUADAMUZ, “Artificial Intelligence and Copyright”, WIPO Magazine (2017), online: WIPO https:// www.wipo.int/wipo_magazine/en/2017/05/article_0003.html#:∼:text=Artificial%20intelligence%20is%20already %20being,used%20and%20reused%20by%20anyone. 47 Section 2(d) of the Indian Copyright Act, 1957, No. 14, Acts of Parliament, 1957. B. What is the Concept of an “Author” in Common Law Countries? The court emphasized the need for a work to exhibit a certain level of permanence and fixation in order to qualify for copyright protection. Furthermore, it is worth noting that the US Copyright Office recently rejected a request for copyright protection for a work titled “A Recent Entrance to Paradise”. The decision by the Copyright Office’s board affirmed a previous ruling, citing the absence of human authorship as a key factor in denying the copyright claim.39 In the evolving landscape of copyright law and AI, several notable litigations have emerged that underscore the complexities of this intersection. A prominent case is the lawsuit initiated by Getty Images against Stability AI, where the former accused the latter of misusing copyrighted photos to train its AI systems.40 This litigation has broader impli- cations, as highlighted by a report which noted that Stability AI allegedly copied at least 12 million copyrighted images from Getty Images for its Generative AI models.41 Parallel to this, there have been significant legal challenges in the UK, with Getty Images seeking to halt the sales of Stability AI’s system, accusing them of using copyrighted images to “train” its Stable Diffusion system.42 Furthermore, a federal judge in the US dismissed an attempt to copyright artwork produced by an AI image generator, emphasizing the intricate nature of authorship and copyright in the age of AI.43 These cases highlight the ongoing debates and challenges in defining copyright boundaries in the realm of Generative AI. Based on the established case law and the current practices of the US Copyright Office, it is evident that claiming or registering independently generated AI works is not permis- sible. The legal framework maintains that creative works produced solely by AI machines are not eligible for copyright protection unless they meet the criteria of being created by https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 10 a human author. The settled case law and the guidelines set forth by the Copyright Office reinforce the principle that copyright law is designed to safeguard the originality and cre- ative contributions of human authors. While AI technology has advanced significantly and is capable of producing impressive works, the absence of direct human involvement in the creative process raises questions regarding the applicability of copyright protection. C. The Dilemma of AI Authorship under Asian Law In the case of an artistic work other than a photograph, the “author” is considered to be the artist. Additionally, the Act recognizes the person taking the photograph as the “author” in the case of photographs. In the context of copyright law, it is interesting to note the contrasting approaches taken by Australia and Thailand. Unlike some jurisdictions, these countries currently lack sufficient protection for works created by AI. This deficiency arises due to the https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 11 Asian Journal of International Law requirement of human personality for copyrightability, which is not fulfilled by AI-generated works. While such works may demonstrate the necessary originality, critics argue that they fail to exhibit the collective attributes of effort, knowledge, and intellect that contribute to the concept of originality.48 In a similar vein, Singapore’s copyright law places the requirement on human authors rather than machines, reflecting its preference for human involvement. This was evident in the case of Asia Pacific Publishing Pte Ltd v Pioneers & Leaders (Publishers) Pte Ltd, where the Singapore Court of Appeal delineated four essential characteristics that determine copy- right eligibility. Initially, section 2(7)49 of the Act did not clearly define who could qualify as an author of a subsisting copyright under the Act. However, the absence of such a def- inition did not imply that the concept of a “qualified person” could be extended to include non-living entities such as incorporated bodies. Moreover, it was evident that the histor- ical context of the act envisioned granting rights exclusively to natural persons rather than corporate bodies, emphasizing that legal rights derive solely from human author- ship.50 This historical perspective further reinforced the notion that originality was closely tied to human authors. Therefore, the identification of an author was a prerequis- ite before determining whether a work could be considered original. Without establishing the human author from whom the work originated, it would be impossible to classify it as an “original work” eligible for copyright protection. Interestingly, this issue was raised and discussed in the case of B2C2 Ltd v Quoine Pte Ltd.51 The Singapore International Commercial Court was tasked with addressing issues pertaining to AI in the context of cryptocurrency. The central question at hand was how legal principles should be applied to a cryptocurrency transaction that was entirely executed through AI algorithms. 48 Nattapong SUWAN-IN, “Copyright Protection on AI-Generated Work: The Case Study of the US, UK, and Thailand Copyright Laws”, (2021) Journal of Law Public Administration and Social Science 131, 142. See also the Australian Copyright Act 1968 (Cth) (Copyright Act) and IceTV Pty Ltd v Nine Network Australia Pty Ltd [2009] HCA 14, where the High Court of Australia clarified that factual data contained in a compilation is not sufficient to attract protection under the Copyright Act. 49 Asia Pacific Publishing Pte Ltd v Pioneers & Leaders (Publishers) Pte Ltd [2011] SGCA 37. 50 The 1911 Act was enacted in a different era where the concept of copyright was in its infancy. The 1911 Act was primarily concerned with the rights of authors, and it is clear from the historical context that the Act was intended to protect the rights of individual authors rather than corporate entities. The emphasis was on human creativity and the personal rights of the author. 51 B2C2 Ltd V Quoine Pte Ltd [2019] SGHC(I) 03. 52 Ibid., at paras. 206–8. 48 Nattapong SUWAN-IN, “Copyright Protection on AI-Generated Work: The Case Study of the US, UK, and Thailand Copyright Laws”, (2021) Journal of Law Public Administration and Social Science 131, 142. See also the Australian Copyright Act 1968 (Cth) (Copyright Act) and IceTV Pty Ltd v Nine Network Australia Pty Ltd [2009] HCA 14, where the High Court of Australia clarified that factual data contained in a compilation is not sufficient to attract protection under the Copyright Act. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press f g 50 The 1911 Act was enacted in a different era where the concept of copyright was in its infancy. The 1911 Act was primarily concerned with the rights of authors, and it is clear from the historical context that the Act was intended to protect the rights of individual authors rather than corporate entities. The emphasis was on human creativity and the personal rights of the author. 51 B2C2 Ltd V Quoine Pte Ltd [2019] SGHC(I) 03. 52 Ibid., at paras. 206–8. D. Should Copyright Law Extend Protection to AI-Generated Works or Exclude Them? The World Intellectual Property Office (WIPO) released a discussion paper titled “Conversation on Intellectual Property Policy and Artificial Intelligence”,53 which addresses the increasing utilization of AI in generating literary and artistic works. The rapid growth of AI capabilities in this domain raises significant policy considerations within the copyright system, which has long been intertwined with the human creative spirit and the recognition of and incentives for human expression. Determining the appropriate legal framework for attributing copyright to AI-generated works raises funda- mental questions that touch upon the fundamental objectives of the copyright system. The copyright system traditionally aimed to safeguard and foster human creativity. Therefore, any decisions regarding the eligibility of AI-generated works for copyright pro- tection would significantly impact the underlying social purpose for which the copyright system was established. If AI-generated works were excluded from copyright protection, it would reinforce the notion that copyright law exists to uphold the dignity and preference given to human over machine-generated creativity. Such an approach would highlight the belief that the copyright system is designed to incentivize and reward human creative efforts. On the other hand, if copyright protection is extended to works created by AI, the copyright system would be perceived as favoring the proliferation of creative works in the hands of consumers, regardless of whether they were generated by humans or machines. This perspective implies that the copyright system prioritizes the availability and accessibility of creative works without making a distinction between human and machine-generated creativity.54 g y During the Third Session on Intellectual Property and Artificial Intelligence, the discus- sions revolved around two opposing perspectives on the issue. The opponents argued against providing copyright protection to works created by AI, basing their opposition on several grounds.55 They highlighted the technical nature of human input combined with the automated processes of AI algorithms, asserting that it does not sufficiently jus- tify copyright protection for AI-generated works. They emphasized that the principle of human authorship is firmly established in copyright law, while the concept of AI is still evolving and lacks clarity. Furthermore, the opponents contended that works produced by AI should be considered part of the public domain.56 They asserted that expanding the scope of AI-assisted outputs, including the arrangement, creation, and selection stages, would raise policy concerns and potentially diminish the public domain. 53 WIPO Secretariat, supra note 4 at para. 23. 54 Ibid. 55 Ibid. 56 Ibid. 57 Ibid. C. The Dilemma of AI Authorship under Asian Law In this regard, Judge Simon Thorley, in his opinion, provided insightful observations on the matter:52 Turning to the issue of knowledge attribution, the legal framework for determining knowledge in cases where computers have replaced human actions is expected to evolve as legal disputes arise from such circumstances. This is especially relevant in situations where the computer involved is equipped with artificial intelligence, potentially possessing a degree of autonomy. Given these complexities, I do not intend to express any specific opinions regarding the precise legal relationship between computers and those who control or program them. In the present case, the algorithmic programs in question are deterministic, meaning they strictly adhere to the instructions programmed into them. They function in a predetermined man- ner when called upon and lack independent decision-making capabilities. They do not possess their own consciousness or understanding of why they perform certain actions, nor are they aware of the external events that trigger their operations. It is worth noting that as disputes arise involving AI-driven systems, the legal https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 12 understanding of knowledge attribution in relation to computer actions will likely develop further. The unique nature of AI and its impact on legal principles will necessitate ongoing examination and refinement within the legal domain. Based on the cases discussed, it can be inferred that Singaporean law does not currently offer explicit protection for literary and artistic works generated by AI. The cases above illustrate a lack of legal provisions and a reluctance to extend copyright protection to works solely created by AI systems. 60 Ryan ABBOTT, “Artificial Intelligence, Big Data and Intellectual Property: Protecting Computer-Generated Works in the United Kingdom” in Tanya APLIN, ed., Research Handbook on Intellectual Property and Digital Technologies, (Edward Elgar Publishing Ltd, Forthcoming), online: Social Science Research Network https://ssrn. com/abstract=3064213. D. Should Copyright Law Extend Protection to AI-Generated Works or Exclude Them? The oppo- nents argued that the public’s access to such creations could be restricted by subjecting more AI-generated literary and artistic works to copyright law.57 https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 13 Asian Journal of International Law Conversely, some scholars have presented arguments advocating for the protection of AI-generated works, asserting that denying such protection due to the absence of human intervention would result in these works falling into the public domain.58 This, in turn, would allow consumers to access and use these works without the fear of copyright infringement claims. They contend that refusing authorship to these categories of works would be detrimental because it would stifle and impede creativity.59 Moreover, granting patent protection to the entity or programmer who created the software used by AI is seen as contradictory when protection is denied to the outputs produced by that software.60 These viewpoints led to the conclusion that AI works should be eligible for copyright protection, regardless of their nature.61 The argument highlights the importance of recognizing and rewarding the creative elements and outputs generated by AI systems.62 By extending copyright protection to AI-generated works, it is believed that a balanced approach can be achieved, fostering both innovation and access to creative works.63 This perspective emphasizes the need to adapt copyright law to accommodate advancements in AI technology and ensure that appropriate legal frameworks are in place to support and incentivize AI-driven creativity.64 58 Jie ZHANG and Xin XIE, “Research on the Copyright Protection of Artificial Intelligence Generation in the Smart Media Environment”, 2023 3rd International Conference on Public Management and Intelligent Society (PMIS 2023) 665. 61 Ibid. 62 Eloghene E. ADAKA and Ifeoluwa A. OLUBIYI, “Lessons for Nigeria: Determining Authorship and Inventorship of Artificial Intelligence Generated Works”, (2022) 2 Journal of Intellectual Property and Information Technology Law 15. gy 63 Sik Cheng PENG, “Artificial Intelligence and Copyright: The Authors’ Conundrum”, (2018) 9 WIPO-WTO Colloquium Papers 173, online: WTO https://www.wto.org/english/tratop_e/trips_e/colloquium_papers_e/ 2018/chapter_13_2018_e.pdf/ 64 b d l l h h ll l d h k l d k Ibid. 62 Eloghene E. ADAKA and Ifeoluwa A. OLUBIYI, “Lessons for Nigeria: Determining Authorship and Inventorship of Artificial Intelligence Generated Works”, (2022) 2 Journal of Intellectual Property and Information Technology Law 15. 63 Sik Cheng PENG, “Artificial Intelligence and Copyright: The Authors’ Conundrum”, (2018) 9 WIPO-WTO Colloquium Papers 173, online: WTO https://www.wto.org/english/tratop_e/trips_e/colloquium_papers_e/ 2018/chapter_13_2018_e.pdf/ 64 Ibid. See also Saleh AL-SHARIEH, “The intellectual property road to the knowledge economy: Remarks on the readiness of the UAE Copyright Act to drive AI innovation”, (2020) 13 Law, Innovation and Technology 141. Inventorship of Artificial Intelligence Generated Works , (2022) 2 Journal of Intellectual Property and Information Technology Law 15. 63 Sik Cheng PENG, “Artificial Intelligence and Copyright: The Authors’ Conundrum”, (2018) 9 WIPO-WTO Colloquium Papers 173, online: WTO https://www.wto.org/english/tratop_e/trips_e/colloquium_papers_e/ 2018/chapter_13_2018_e.pdf/ 64 Ibid See also Saleh AL SHARIEH “The intellectual property road to the knowledge economy: Remarks on 66 WIPO Secretariat, supra note 4 at para. 23. 67 Ibid. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 66 WIPO Secretariat, supra note 4 at para. 23. 58 Jie ZHANG and Xin XIE, “Research on the Copyright Protection of Artificial Intelligence Generation in the Smart Media Environment”, 2023 3rd International Conference on Public Management and Intelligent Society (PMIS 2023) 665. 59 Ibid. 60 Ryan ABBOTT, “Artificial Intelligence, Big Data and Intellectual Property: Protecting Computer-Generated Works in the United Kingdom” in Tanya APLIN, ed., Research Handbook on Intellectual Property and Digital Technologies, (Edward Elgar Publishing Ltd, Forthcoming), online: Social Science Research Network https://ssrn. com/abstract=3064213. 61 Ibid. 62 Eloghene E. ADAKA and Ifeoluwa A. OLUBIYI, “Lessons for Nigeria: Determining Authorship and Inventorship of Artificial Intelligence Generated Works”, (2022) 2 Journal of Intellectual Property and Information Technology Law 15. 63 Sik Cheng PENG, “Artificial Intelligence and Copyright: The Authors’ Conundrum”, (2018) 9 WIPO-WTO Colloquium Papers 173, online: WTO https://www.wto.org/english/tratop_e/trips_e/colloquium_papers_e/ 2018/chapter_13_2018_e.pdf/ 64 Ibid. See also Saleh AL-SHARIEH, “The intellectual property road to the knowledge economy: Remarks on the readiness of the UAE Copyright Act to drive AI innovation”, (2020) 13 Law, Innovation and Technology 141. 65 Ibid. 66 WIPO Secretariat, supra note 4 at para. 23. 67 Ibid. III. Ownership of Copyright in AI-Generated Works: Attribution and Vesting Considerations In the context of works produced by AI applications, the term “author” is generally understood as referring to the creator or maker of a work.65 Hence, the debate surround- ing the author’s identification in the case of AI-generated works may seem inconsequen- tial; these works ultimately stem from human creativity. Therefore, it can be contended that AI should be regarded as a tool or instrument employed by the human author rather than an autonomous creator in its own right.66 When considering the question of copy- right ownership, some scholars have proposed three possible candidates. The first nom- inee is the human author who made the necessary arrangements for the creation of the work, particularly in the case of AI-assisted works.67 The suggestion of attributing copy- right to a human author is rooted in the idea that establishing a causal connection between the input of the human author and the resulting AI-generated work can serve as a significant factor in determining authorship. This approach becomes especially 58 Jie ZHANG and Xin XIE, “Research on the Copyright Protection of Artificial Intelligence Generation in the Smart Media Environment”, 2023 3rd International Conference on Public Management and Intelligent Society (PMIS 2023) 665. 59 62 Eloghene E. ADAKA and Ifeoluwa A. OLUBIYI, “Lessons for Nigeria: Determining Authorship and Inventorship of Artificial Intelligence Generated Works”, (2022) 2 Journal of Intellectual Property and Information Technology Law 15. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 14 valuable when multiple individuals are involved in the creative process; it aids in deter- mining the true relationship between the contributors and deciding issues of authorship or co-authorship of the work.68 The approach to authorship aligns with the statutory definitions found in various jur- isdictions. For instance, in the UK, section 9(3) of the Copyright, Designs and Patents Act (CDPA) of 1988 stipulates that in the case of a computer-generated literary, dramatic, musical, or artistic work, the person who undertakes the necessary arrangements for the creation of the work is considered to be the author.69 Similarly, section 178 of the CDPA defines AI-generated works as those produced by a computer in circumstances where no human author is involved. Ibid. 69 Andrés GUADAMUZ, “Do Androids Dream of Electric Copyright? Comparative Analysis of Originality in Artificial Intelligence Generated Works”, (2017) 2 Intellectual Property Quarterly 169. 70 f h l d h ( ) III. Ownership of Copyright in AI-Generated Works: Attribution and Vesting Considerations This perspective is also reflected in the legislation of other countries such as New Zealand,70 Hong Kong,71 and Ireland,72 where the person who makes the essential arrangements for the creation of a computer-generated work is considered to be the author. The copyright laws of the UK and similar jurisdictions established a legal framework that addressed the protection of AI-generated works in cases where there is no identifi- able human author. This legal framework recognizes the significance of the individual who undertakes the essential steps for the creation of such works. As a result, it can be inferred that copyright protection for AI-generated works is contingent upon a sub- stantial contribution made by the person responsible for orchestrating the necessary pro- cesses involved in their creation. These legal provisions acknowledge the importance of human involvement in the creative process, even in cases where the final output is gen- erated by AI or computer algorithms. y g The second contender for copyright ownership is the technical program itself, particu- larly in cases where the work is solely generated by AI. Advocates of this viewpoint argue that it diverges from the previous scenario where human input played a role. In the con- text of AI applications, a unique situation exists wherein a computer, utilizing AI capabil- ities, autonomously generates literary or artistic works without direct human intervention. This apparent gap between human input and the computer’s output creates the possibility of considering the AI program itself as a candidate for copyright owner- ship.73 Regarding the previous hypothesis, it has been contended that the user’s contribu- tion alone may not be adequate to trace the complete creative process of a work generated by AI. Consequently, it has been suggested that, in such cases, the individual responsible for programming the intrinsic elements of the work should be regarded as 69 Andrés GUADAMUZ, “Do Androids Dream of Electric Copyright? Comparative Analysis of Originality in Artificial Intelligence Generated Works”, (2017) 2 Intellectual Property Quarterly 169. 70 f h l d h ( ) g (1) For the purposes of this Act, the author of a work is the person who creates it. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 68 Ibid. 68 Ibid. 69 Andrés GUADAMUZ, “Do Androids Dream of Electric Copyright? Comparative Analysis of Originality in Artificial Intelligence Generated Works”, (2017) 2 Intellectual Property Quarterly 169. 70 Section 5 of the New Zealand Copyright Act (1994) states: 74 Peng, supra note 63 at 181–2. 75 Robert C. DENICOLA, “Ex Machina: Copyright Protection for Computer-Generated Works”, (2016) 69 Rutgers U. L. Rev. 251, at 280–1. 76 Darin GLASSER, “Copyrights in Computer-Generated Works: Whom, if Anyone, Do We Reward?” (2001) 1 Duke Law & Technology Review 1. 77 C.J.S. AZORO and Queen O. AGULEFO, “‘Original’ under the law of copyright is distinct from the ordinary meaning of ‘original’: A discourse”, (2021) 7 International Journal of Law 29, 30–1. 78 Alexander D. NORTHOVER, “Enough and as Good in the Intellectual Commons: A Lockean Theory of Copyright and the Merger Doctrine”, (2016) 65 Emory L. J. 1,363, at 1,376–7. III. Ownership of Copyright in AI-Generated Works: Attribution and Vesting Considerations (2) For the purposes of sub-section (1), the person who creates a work shall be taken to be: (a) in the case of a literary, dramatic, musical, or artistic work that is computer-generated, the person by whom the arrangements necessary for the creation of the work are undertaken”. ( ) p p ( ), p (a) in the case of a literary, dramatic, musical, or artistic work that is computer-generated, the person by whom the arrangements necessary for the creation of the work are undertaken”. 71 Section 11(3) of Copyright Ordinance (CO) (Cap. 528), states: “In the case of a literary, dramatic, musical or artistic work which is computer-generated, the author is taken to be the person by whom the arrangements necessary for the creation of the work are undertaken”. 71 Section 11(3) of Copyright Ordinance (CO) (Cap. 528), states: “In the case of a literary, dramatic, musical or artistic work which is computer-generated, the author is taken to be the person by whom the arrangements necessary for the creation of the work are undertaken”. 72 Section 21( f ) of Copyright and Related Rights Act (2000) states: “In this Act, ‘author’ means the person who creates a work and includes: in the case of a work which is computer-generated, the person by whom the arrangements necessary for the creation of the work are undertaken.” 72 Section 21( f ) of Copyright and Related Rights Act (2000) states: “In this Act, ‘author’ means the person who creates a work and includes: in the case of a work which is computer-generated, the person by whom the arrangements necessary for the creation of the work are undertaken.” 73 Nahide BASRI, “The Question of Authorship in Computer-Generated Work”, Journal of Law & Social Change (13 January 2020), online: Penn Carey Law https://www.law.upenn.edu/live/news/9691-the-question-ofauthorship- in-computer-generated. 73 Nahide BASRI, “The Question of Authorship in Computer-Generated Work”, Journal of Law & Social Change (13 January 2020), online: Penn Carey Law https://www.law.upenn.edu/live/news/9691-the-question-ofauthorship- in-computer-generated. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Asian Journal of International Law 15 the author. This perspective highlights the essential role played by the person who devel- ops the programming that enables the production of the specific work. Furthermore, it has been argued that copyright ownership should be vested in the entity or programmer who developed the AI application itself. III. Ownership of Copyright in AI-Generated Works: Attribution and Vesting Considerations By recognizing the entity or programmer as the copyright holder, this approach acknowledges the overarching creative input involved in designing and implementing the AI system.74 It emphasizes that copyright protection should be attributed to those responsible for developing and establishing the AI applica- tion that ultimately generates the work.75 Initially, it was suggested that the person responsible for making the necessary arrangements, particularly in the case of AI-assisted works, where AI serves as a creative tool or aids in the work’s production, should be recognized as the author of the work and thus entitled to copyright ownership. The rationale behind this argument is that it would be unrealistic to expect a human imprint on every aspect of AI-assisted works. Instead, the focus should be on the min- imum level of contribution required to meet the criterion of originality. It is important to consider that assigning a copyright solely based on a human identity would not adequately address the complexities of works solely generated by AI. Merely attributing authorship to a human without direct creative input may not capture the true nature of the work. In such cases, the conventional notion of authorship tied to human identity may not sufficiently reflect the abstract reality of AI-generated works.76 When approach- ing the question of authorship within the context of AI-generated works, it becomes apparent that it is a complex issue, and ascribing traditional notions of authorship in this scenario may present significant challenges. However, a more pragmatic solution could involve assigning ownership of the produced work to the owner of the AI program itself. 74 Peng, supra note 63 at 181–2. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press IV. Can AI-Generatyed Works Be Regarded as Original If Copyright is Attributed? The criterion of originality plays a crucial role in copyright law, serving as a yardstick to determine whether a specific work is eligible for copyright protection. This term is com- monly employed to differentiate between works that possess originality and those that do not. Essentially, “originality” signifies that a work deserving of protection must originate from an identifiable creator or author, thereby reflecting the author’s distinctive person- ality rather than being wholly derivative or entirely new.77 y g y y Under the common law, the concept of originality has historically been associated with the “sweat of the brow” theory.78 However, it is important to note that in countries such as the US, the criterion for originality has evolved, and the mere labour employed in the creation of a work is no longer a requisite for establishing originality or for qualifying a work for copyright protection. This theory aligns with the utilitarian approach that pro- motes human creativity and incentivizes authors to gain copyright protection for their works. According to this approach, authors can acquire copyright through their diligent efforts during the creation of a work, whether it be a database, a collection of literary https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 16 works, or an artistic compilation such as an encyclopedia.79 The case of Hollinrake v Truswell80 further clarified the concept of originality in copyright law. It established that originality is not contingent upon the novelty or uniqueness of a work but rather on its origin from the author. In other words, for a work to be deemed original, it must originate from the author’s own creative efforts and not be copied from an external source. This ruling reaffirmed that originality focuses on the independent creation of a work by the author rather than its level of novelty. By emphasizing the requirement of authorship and the absence of copying, the decision in Hollinrake v Truswell provided clar- ity on the understanding of originality within the jurisdiction of England and Wales. In the case of University of London Press v University Tutorial Press,81 the court highlighted that a work is considered original when it demonstrates a certain level of skill and labour and is not copied from another existing work. This understanding of originality, which focuses on independent creation and the absence of copying, has been influential in the UK. IV. Can AI-Generatyed Works Be Regarded as Original If Copyright is Attributed? It is important to note that the standard of skill and labour applied in the UK has been significantly shaped by the perspective of the CJEU.82 g y y Similar to the UK, the courts in the US have long recognized that originality requires a minimal level of creative effort. In the case of Burrow-Giles Lithographic Co. v Sarony, the court emphasized that copyrightability necessitates a certain amount of creative labour. Similarly, in Time Inc. v Bernard Geis Associates, a district court in the US reached a similar conclusion. The court referred to the standard of originality and acknowledged the pres- ence of an element of personality in the creation of a photograph, such as the choice of subject, framing, and timing. The court recognized the visual appeal and artistic nature of a photograph but refrained from acting as a critic by evaluating the artistic merit, skill, or effort involved in the photograph.83 In the case of Baltimore Orioles v MLB Players Association, the US Court of Appeals for the Seventh Circuit addressed an appeal stemming from a pro- longed dispute between major league baseball clubs (referred to as “Clubs”) and the Major League Baseball Players Association. The court provided clarity on the threshold of origin- ality, asserting that it encompasses two distinct conditions. These conditions are that a work must possess an independent origin and exhibit a minimal level of creativity. By dis- tinguishing between independent origin and minimal creativity, the court emphasized the dual requirements that must be met for a work to satisfy the threshold of originality. The concept of independent origin implies that the work must originate from the author’s own creation rather than being a copy or imitation of existing material. Additionally, the concept of minimal creativity acknowledges that even a small degree of creative effort is necessary for a work to meet the originality requirement84. In the case of Alfred Bell & Co. v Catalda Fine Arts Inc., the Court of Appeal provided further clarity on the concept of ori- ginality. It explicitly stated that originality requires two essential elements. First, the work must have originated from the author, emphasizing the need for independent creation. Second, the work must possess at least a minimal level of novelty, indicating that it should exhibit some degree of newness or uniqueness. 79 Azoro and Agulefo, supra note 77 at 31. 80 Hollinrake v Truswell [1894] 3. Ch. 420, para. 427. 81 University of London Press v University Tutorial Press [1916] 2 Ch 601. 82 The CJEU has delivered a series of rulings concerning the test of originality, where the realization of a sub- ject matter has been dictated by technical considerations, rules or other constraints which left no room for cre- ative freedom. (See Newspaper Licensing Agency Ltd v Meltwater Holding BV [2011] EWCA Civ 890 detailing an equal criterion of assessing originality for other types of works. 83 Time Inc. v Bernard Geis Associates, 293 F. Supp. 130 (1968). 84 Baltimore. Orioles, Inc. v Major League Baseball Players Ass’n, 805 F.2d 663 (7th Cir. 1986). 84 Baltimore. Orioles, Inc. v Major League Baseball Players Ass’n, 805 F.2d 663 (7th Cir. 1986). 83 Time Inc. v Bernard Geis Associates, 293 F. Supp. 130 (1968). https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press IV. Can AI-Generatyed Works Be Regarded as Original If Copyright is Attributed? By articulating the requirement of inde- pendent origin and the presence of even the slightest level of novelty, the court shed light on the fundamental aspects of originality. This ruling held that a work must not be a mere https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 17 Asian Journal of International Law copy or imitation but must possess an element of freshness or distinctiveness in order to meet the originality threshold.85 In the landmark case of Feist Publications, Inc. v Rural Telephone Service Co., the US Supreme Court clarified the requirements for a work to be eligible for copyright protection. The Court established that for a work to be entitled to copyright, it must satisfy two crucial conditions. First, the work must be created by the author, emphasizing the importance of independent creation. Second, the work must exhibit a minimum level of creativity. The Supreme Court’s ruling emphasized that originality in copyright law necessitates a modest degree of intellectual labour. This means that the work must embody a certain amount of creative effort or originality, indicating that it goes beyond mere facts or data.86 The decision in Feist Publications, Inc. v Rural Telephone Service Co. provides significant guidance on the originality standard, affirm- ing that copyright protection is granted to works that demonstrate a genuine level of intellectual contribution.87 The consideration of two opposing views, namely the subject- ive and objective theories, and their impact on the test of originality for works created by technical means is crucial. However, neither of these theories has proven to be an effect- ive sole criterion for determining the threshold of originality. The subjective theory emphasizes the individual author’s subjective creativity and personal expression as the basis for originality. Strictly applying this theory to AI-generated works, however, could exclude them from copyright protection, undermining the purpose of the copyright system.88 By contrast, the objective theory relies on objective criteria such as skill, effort, or labour invested in the work. A purely objective approach may risk attributing copyright to entities or individuals who did not contribute creatively to the work, thus assigning ownership to the wrong person. To address these shortcomings, both common law and civil law jurisdictions recognize the need for a balanced approach. They are moving towards a middle ground that acknowledges the complexities of originality in the context of AI-generated works. 85 Alfred Bell & Co. Ltd. v Catalda Fine Arts, Inc. et al., 191 F.2d 99 (2d Cir. 1951). 86 Feist Publications, Inc. v Rural Telephone Service Co., Inc., 499 U.S. 340 (1991). 87 Burrow-Giles Lithographic Co. v Sarony, 111, U.S. 53 (1884). 88 Zhe DAI and Banggui JIN, “The copyright protection of AI-generated works under Chinese law” (2023) 13 Juridical Tribune 241. 89 According to the arguments of German philosophers Kant and Hegel specifically in 1875, the idea of copy- right ownership is based on the personal theory, which goes on to say that property is an expression of human personality. Hegel notes that will, freedom, and the person are the most important parts of every personality. See Christopher YOO, “Rethinking Copyright and Personhood” (2019) 2019 University of Illinois Law Review 1,049. f 86 Feist Publications, Inc. v Rural Telephone Service Co., Inc., 499 U.S. 340 (1991). p 87 Burrow-Giles Lithographic Co. v Sarony, 111, U.S. 53 (1884). 85 Alfred Bell & Co. Ltd. v Catalda Fine Arts, Inc. et al., 191 F.2d 99 (2d Cir. 1951). 89 According to the arguments of German philosophers Kant and Hegel specifically in 1875, the idea of copy- right ownership is based on the personal theory, which goes on to say that property is an expression of human personality. Hegel notes that will, freedom, and the person are the most important parts of every personality. See Christopher YOO, “Rethinking Copyright and Personhood” (2019) 2019 University of Illinois Law Review 1,049. 85 Alfred Bell & Co. Ltd. v Catalda Fine Arts, Inc. et al., 191 F.2d 99 (2d Cir. 1951). 86 Feist Publications, Inc. v Rural Telephone Service Co., Inc., 499 U.S. 340 (1991). g p y 88 Zhe DAI and Banggui JIN, “The copyright protection of AI-generated works under Chinese law” (2023) ridical Tribune 241. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press IV. Can AI-Generatyed Works Be Regarded as Original If Copyright is Attributed? Instead of solely focusing on novelty, these jurisdictions seek to ascertain whether a work is created by the author and not copied from others. Common law countries such as the UK and the US prioritize authorship over novelty in their approach to originality. By avoiding the extremes of subjective and objective theor- ies, these jurisdictions seek to maintain a fair and functional copyright system that accounts for the unique characteristics of AI-generated works. In contrast to common law jurisdictions, civil law countries approach the question of originality from a perspective that often draws on case law, especially within the EU. They place less emphasis on rewarding the labour and skill invested in creating a work and more on the inherent connection between the author and the creation. Within civil law systems, originality is often viewed through the lens of an unbreakable link of paternity, highlighting the importance of the author’s personal connection to the work. Moral rights derive from subjective theory and play a significant role in civil law countries. These rights include the author’s entitlement to be credited for their work and to maintain con- trol over its integrity. While the subjective approach underlies these moral rights,89 oppo- nents argue that it may not adequately address the unique nature of AI-generated 89 According to the arguments of German philosophers Kant and Hegel specifically in 1875, the idea of copy- right ownership is based on the personal theory, which goes on to say that property is an expression of human personality. Hegel notes that will, freedom, and the person are the most important parts of every personality. See Christopher YOO, “Rethinking Copyright and Personhood” (2019) 2019 University of Illinois Law Review 1,049. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 18 works.90 In the context of AI-generated works, the application of a purely subjective per- spective may seem unclear and ambiguous. As AI systems autonomously produce works without direct human intervention, the traditional criteria for originality based on the author’s personality become less applicable. This raises concerns about whether AI-generated works can meet the requirements for copyright protection under a strictly individualistic approach.91 In conclusion, the search for an effective test of originality in the realm of AI-generated works requires a nuanced understanding. Jurisdictions are mov- ing towards a balanced approach that acknowledges authorship while considering the spe- cific challenges posed by AI-generated works. IV. Can AI-Generatyed Works Be Regarded as Original If Copyright is Attributed? The challenges surrounding originality in the context of AI-generated works have prompted French courts to transition from a rigid standard of personality to a more adaptable and flexible approach. To determine how an author expresses their personality through a work, the courts have focused on the question of what actions the author has taken to showcase their individuality. A pre- vailing response has emerged through judicial practice, highlighting that creative choices play a pivotal role in demonstrating the author’s personality.92 90 K. I. ADAM and I. A. YUSUF, “Originality in Copyright and the Debate on Protection of Traditional Knowledge: A View on Nigerian Law”, (2017) 3 KIU Journal of Social Sciences 293, at 297. 91 Ibid. 93 Berne Convention, supra note 24, Art. 2(3). 94 Ibid., Art. 2(5). https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 90 K. I. ADAM and I. A. YUSUF, “Originality in Copyright and the Debate on Protection of Traditional Knowledge: A View on Nigerian Law”, (2017) 3 KIU Journal of Social Sciences 293, at 297. 91 Ibid. 92 Ibid. 93 Berne Convention supra note 24 Art 2(3) 95 Examples provided under Article 2(1) of the Berne Convention include “books, pamphlets and other writ- ings; lectures, addresses, sermons and other works of the same nature; dramatic or dramatic-musical works; choreographic works and entertainments in dumb show; musical compositions with or without words; cinemato- graphic works to which are assimilated works expressed by a process analogous to cinematography; works of 90 K. I. ADAM and I. A. YUSUF, “Originality in Copyright and the Debate on Protection of Traditional Knowledge: A View on Nigerian Law”, (2017) 3 KIU Journal of Social Sciences 293, at 297. 91 Ibid. 92 Ibid. 93 Berne Convention, supra note 24, Art. 2(3). 94 Ibid., Art. 2(5). 95 Examples provided under Article 2(1) of the Berne Convention include “books, pamphlets and other writ- ings; lectures, addresses, sermons and other works of the same nature; dramatic or dramatic-musical works; choreographic works and entertainments in dumb show; musical compositions with or without words; cinemato- graphic works to which are assimilated works expressed by a process analogous to cinematography; works of V. The Threshold of Originality in International Copyright Law Exploring the concept at an international level is essential before delving into the analysis of originality as a requirement in AI-generated works. The Berne Convention serves as a relevant reference point, addressing the protection of works and the rights of their authors. While the term “originality” is not explicitly mentioned, it can be inferred from Article 2(3), which states that translations, adaptations, musical arrangements, and other alterations of a literary or artistic work shall be protected as original works.93 This provision suggests that these derivative works are entitled to independent copyright protection, implying an underlying requirement of originality. The concept can also be inferred from Article 2(5), which addresses collections of literary or artistic works, such as encyclopedias and anthologies. It states that these collections, by virtue of the selection and arrangement of their contents, qualify as intellectual creations. Article 2 (5) recognizes that the compilation and arrangement require intellectual creativity and effort, granting them protection separate from the copyrights of the individual works they comprise.94 In conclusion, the Berne Convention underscores the importance of copyright protection for works that embody intellectual creation, emphasizing eligibility as long as they demonstrate their originator’s intellectual creativity and originality. The convention affirms the eligibility of collections for copyright protection, acknowledg- ing the compilation itself as an original creation resulting from the intellectual choices and efforts of the compiler or editor. p Article 2(1) of the Berne Convention is a relevant reference point to determine whether AI-generated works meet the requirement of originality. This provision states that literary and artistic works encompass all creations in the literary, scientific, and artistic domain, regardless of their mode or form of expression.95 Consequently, while the Berne 95 Examples provided under Article 2(1) of the Berne Convention include “books, pamphlets and other writ- ings; lectures, addresses, sermons and other works of the same nature; dramatic or dramatic-musical works; choreographic works and entertainments in dumb show; musical compositions with or without words; cinemato- graphic works to which are assimilated works expressed by a process analogous to cinematography; works of https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 19 Asian Journal of International Law Convention emphasizes the importance of originality for copyright protection, it does not specifically address the eligibility of AI-generated outputs; such works were not contemplated at the time of its drafting. 96 Berne Convention, supra note 24, Art. 2(2). 97 Thomas MARGONI, “The Harmonisation of EU Copyright Law: The Originality Standard”, (30 June 2016) online: Social Science Research Network https://ssrn.com/abstract=2802327. 98 See, Directive 91/250/EEC (Software Directive); Directive 2009/24/EC, Directive 96/9/EC (Database); Directive 2006/116/EC (Copyright Term Directive, with regard to photographs) and Directive 2001/29/EC (Directive on the harmonisation of certain aspects of copyright and related rights in the information society). 99 Hugenholtz and Quintais, supra note 23 at 1,193–4. 100 Case C-5/08, Infopaq International v Danske Dagblades Forening Infopaq International [2009] ECR I-6569. drawing, painting, architecture, sculpture, engraving and lithography; photographic works to which are assimi- lated works expressed by a process analogous to photography; works of applied art, illustrations, maps, plans, sketches and three-dimensional works relative to geography, topography, architecture or science”. See supra note 24. drawing, painting, architecture, sculpture, engraving and lithography; photographic works to which are assimi- lated works expressed by a process analogous to photography; works of applied art, illustrations, maps, plans, sketches and three-dimensional works relative to geography, topography, architecture or science”. See supra note 24. 96 Berne Convention, supra note 24, Art. 2(2). 97 Thomas MARGONI, “The Harmonisation of EU Copyright Law: The Originality Standard”, (30 June 2016) online: Social Science Research Network https://ssrn.com/abstract=2802327. 98 See, Directive 91/250/EEC (Software Directive); Directive 2009/24/EC, Directive 96/9/EC (Database); Directive 2006/116/EC (Copyright Term Directive, with regard to photographs) and Directive 2001/29/EC (Directive on the harmonisation of certain aspects of copyright and related rights in the information society). 99 Hugenholtz and Quintais, supra note 23 at 1,193–4. 100 Case C-5/08, Infopaq International v Danske Dagblades Forening Infopaq International [2009] ECR I-6569. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 96 Berne Convention, supra note 24, Art. 2(2). V. The Threshold of Originality in International Copyright Law However, it is important to note that the con- vention allows for flexibility by permitting national laws to prescribe certain condi- tions for protection. Specifically, national laws may require works, in general or within specified categories, to be fixed in a tangible or material form to be eligible for copyright protection. This provision ensures that works, including AI-generated works, are appropriately safeguarded once they are manifested in a tangible medium.96 Therefore, the specific threshold of originality required for copyright protection in AI-generated works ultimately depends on the laws and regulations of each jurisdiction. Since the Berne Convention grants member countries flexibility to determine the ori- ginality level necessary, the degree of originality required may vary from jurisdiction to jurisdiction. g p 100 Case C-5/08, Infopaq International v Danske Dagblades Forening Infopaq International [2009] ECR I-6569. Berne Convention, supra note 24, Art. 2(2). 97 Thomas MARGONI, “The Harmonisation of EU Copyright Law: The Originality Standard”, (30 June 2016) online: Social Science Research Network https://ssrn.com/abstract=2802327. 98 See, Directive 91/250/EEC (Software Directive); Directive 2009/24/EC, Directive 96/9/EC (Database); VI. The Expansive Interpretation of Originality By the Court of Justice of The European Union (Cjeu) and The Eu’s Approach to the Question of Originality in Copyright https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 20 The author can achieve an original expression through these creative elements, resulting in an intellectual creation.101 Infopaq International A/S v Danske Dagblades Forening is a significant case that sheds light on the complex intersection of copyright infringement and the new technology in emer- ging business models, particularly those involving content aggregation.102 Furthermore, it is worth noting that the European standard of originality, which centres around the author’s intellectual creativity, is not confined solely to the specific categories outlined in legislation, such as software, images, and databases. Instead, this metric is applicable to all types of works falling within the scope of the Berne Convention within the European Union.103 Infopaq International A/S v Danske Dagblades Forening is a significant case that sheds light on the complex intersection of copyright infringement and the new technology in emer- ging business models, particularly those involving content aggregation.102 Furthermore, it In the case of Levola Hengelo BV v Smilde Foods BV,104 the CJEU addressed the issue of copyright protection under Directive 2001/29. The CJEU clarified that for a work to be eli- gible for copyright protection, two cumulative conditions must be satisfied. First, the sub- ject matter in question must be original, meaning that it must be the result of the author’s own intellectual creation105. Second, in accordance with the ruling in Levola Hengelo BV v Smilde Foods BV, the CJEU clarified that for subject matter to be classified as a “work” within the meaning of Directive 2001/29, it must be the expression of the author’s own intellectual creation.106 Similarly, the principle established in Eva-Maria Painer v Standard Verlags GmbH and Others107 aligns with the approach taken in Levola Hengelo BV v Smilde Foods BV. The CJEU was tasked with determining whether a photograph used as a template for a photofit could be considered an original work resulting from the intellectual creation of the appli- cant. The notion of “original work resulting from the intellectual creation” is not expli- citly defined in Directive 93/98 or Directive 2006/116; it is a Union law concept requiring autonomous interpretation. To interpret this notion, reference was made to the relevant recitals in the preamble of Directive 93/98 and Directive 2006/116, which make reference to the Revised Berne Convention. VI. The Expansive Interpretation of Originality By the Court of Justice of The European Union (Cjeu) and The Eu’s Approach to the Question of Originality in Copyright VI. The Expansive Interpretation of Originality By the Court of Justice of The European Union (Cjeu) and The Eu’s Approach to the Question of Originality in Copyright It is essential to recognize that the principle of originality forms the foundation of copy- right protection in national legal systems across Europe. In fact, the absence of this cri- terion would render a work ineligible for copyright protection. Consequently, originality has consistently been employed as a benchmark to determine whether a work should be granted protection97 or deemed non-protected. Despite its fundamental importance, the European Directives98 define originality solely for computer programs, databases, and photographs, describing it as “the author’s own intellectual creation”. It is important to note that EU law does not universally prescribe the criterion of ori- ginality as a prerequisite for legal protection, except in specific cases of computer pro- grams, databases, and photographs. Consequently, originality cannot be regarded as a general rule for determining the eligibility for protection under EU copyright law. However, the European Court of Justice (ECJ) has played a crucial role in shaping the notion of originality and adapting it to address the challenges posed by AI-generated outputs.99 As discussed earlier in this document, the notion of the author’s own intellectual cre- ation has traditionally been limited to certain categories of works. However, the CJEU has played a pivotal role in broadening the application of this standard to encompass a wide range of works, including those generated by AI. In the case of Infopaq International A/S v Danske Dagblades Forening,100 the CJEU established the requirement of originality as the author’s own intellectual creation. The court emphasized that a work must exhibit cre- ativity expressed through the author’s choice, sequence, and combination of words. drawing, painting, architecture, sculpture, engraving and lithography; photographic works to which are assimi- lated works expressed by a process analogous to photography; works of applied art, illustrations, maps, plans, sketches and three-dimensional works relative to geography, topography, architecture or science”. See supra note 24 g p 100 Case C-5/08, Infopaq International v Danske Dagblades Forening Infopaq International [2009] ECR I-6569. 101 Ibid., para. 45. 102 Ibid., para. 36. 103 Ibid., paras. 35–6. 104 Case C-310/17, Levola Hengelo BV v Smilde Foods BV [2018] ECLI:EU:C:2018:899. 105 See also, C 403/08 and C 429/08, EU:C:2011:631, Football Association Premier League and Others, para. 97. 106 Levola, paras. 34–7. 107 C-145/10 Eva-Maria Painer v Standard Verlags GmbH and Others (Third Chamber) [2011] ECR I-12533. 108 Ibid., para. 120. 109 Ibid., para. 121. 110 (C-683/17) ECLI: EU:C: 2019:721, Cofemel v G-Star Raw [Cofemel], para. 31. VII. Copyright Protection for AI-Generated Output in the European Union The ECJ has firmly established that originality is a fundamental criterion for the protec- tion of all types of literary and artistic works under EU law, irrespective of their mode or form of creation. This emphasis on originality highlights its pivotal role in determining the eligibility for copyright protection. However, a closer examination of the ECJ’s case law reveals that this interpretation does not apply universally to all AI-generated works. A notable case that sheds light on the application of originality to AI-generated works is Football Dataco Ltd v Yahoo! UK Ltd.112 In this case, the court acknowledged that a work can be considered an intellectual creation if it reflects the personality of its author. This reflection of personality arises when the author is able to exercise free and creative choices in the production of the work. However, the court also emphasized that if the fea- tures of a work are predetermined by its technical function, the necessary level of origin- ality may be absent.113 y y Another case that further clarifies the concept of originality in the context of AI-generated works is Cofemel v G-Star. In this case, the court emphasized that a subject matter cannot be considered original if its realization has been solely dictated by tech- nical considerations, rules, or other constraints that leave no room for creative freedom. The court highlighted that the presence of creative freedom is essential for a subject mat- ter to meet the required level of originality and qualify as a protected work.114 g y y In the case of Brompton Bicycle Ltd v Chedech/Get2Get,115 the court reiterated the prin- ciple that the realization of a subject matter must not be solely driven by technical con- siderations, rules, or constraints that leave no room for creative freedom. The court emphasized that if a subject matter lacks the necessary creative freedom due to such con- straints, it cannot be considered as possessing the level of originality required to be recog- nized as a protected work116. Based on the cited cases, it may initially seem that the judicial practice tends to deny protection for AI-generated works. However, this interpretation is misleading. In reality, the determination of whether AI-generated works are eligible for copyright protection is not solely based on whether technical considerations dictated their realization. 111 In Football Dataco Ltd and Others v Yahoo! UK Ltd and Others, according to the Opinion of Advocate Genera Mengozzi: “Clearly, it is not possible to define, once and for all and in general terms, what constitutes an ‘intel lectual creation.’ That depends on an assessment which, as I have said, is not necessary in the present case. In any event, if ever that assessment is required, it is for the national courts to undertake it on the basis of the circumstances of each individual case.” See the Opinion of Advocate General Mengozzi, delivered on 15 December 2011, para.38. ECLI: EU: C: 2011:848. 112 Ibid., Football Dataco Ltd, para. 39. 113 See the Opinion of Advocate General Mengozzi in Football Dataco Ltd, supra note 111, para. 35. 114 C-683/17, EU: C: 2019:721, supra note 110 para. 31. 115 Case C-833/18, SI & Brompton Bicycle Ltd. v. Chedech / Get2Get, para. 24. 116 Ibid. 117 Ibid., para. 26. VI. The Expansive Interpretation of Originality By the Court of Justice of The European Union (Cjeu) and The Eu’s Approach to the Question of Originality in Copyright According to the 17th Recital in the Preamble to Directive 93/98 and the 16th Recital in the Preamble to Directive 2006/ 116, an original photographic work exists if it reflects the author’s own intellectual cre- ation and personality. Furthermore, the first sentence of Article 6 of both Directive 93/98 and Directive 2006/116108 establishes that only human creations are eligible for protec- tion, even if technical aids such as cameras are employed in the creative process. This means that works resulting from the application of human creativity, even with the assist- ance of technical tools, can be protected under copyright law.109 Another relevant case that supports the aforementioned approach is Cofemel v G-Star. In this case, the CJEU fur- ther clarified the concept of originality. It stated that, for a subject matter to be consid- ered original, it is both necessary and sufficient that it reflects the personality of its author, representing their free and creative choices. In other words, the subject matter must be the result of the author’s own intellectual creation.110 Based on the analysis of the cited cases, it is important to note that originality is intrinsically linked to the author’s intellectual contribution. However, there is ongoing https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Asian Journal of International Law 21 debate and disagreement regarding the extent of human intervention required to estab- lish originality.111 https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press VII. Copyright Protection for AI-Generated Output in the European Union Instead, the crucial factor is whether the works satisfy the condition of originality and reflect the author’s personality through free and creative choices despite any technical constraints.117 The examples provided highlight that originality is not fulfilled when the subject mat- ter is solely generated by AI applications or predetermined by rules and restrictions with- out room for human creativity. In such cases, fully automated works do not meet the https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 22 criteria for copyright protection according to the settled case law of the CJEU. However, the situation differs for AI-assisted works, where human input and involvement are pre- sent. These works can be considered eligible for copyright protection118 as long as they bear the imprint of human creativity. The human touch and contribution play a vital role in satisfying the requirement of originality. This conclusion is supported by the rationale behind copyright protection, which aims to incentivize authors to create more original and creative works utilizing their unique human abilities.119 Contrary to this view, some jurists argue that the definition of “author” has been heavily influenced by the introduction of AI-generated works. They raise important questions about the pre- vailing definition of the “author” of a computer-generated work, which is often described as “the person by whom the arrangements necessary for the creation of the work are undertaken”. These jurists question whether the EU’s current approach adequately addresses the unique characteristics and challenges posed by AI-generated works. They suggest that the EU should consider revisiting its approach and adapting it to better align with the evolving nature of creativity and authorship in the context of AI.120 Such a reassessment would aim to ensure a more comprehensive and effective framework for protecting and incentivizing innovative AI-generated works while balancing the inter- ests of human creators and technological advancements. Accordingly, the proposed stand- ard broadens the concept of authorship to include individuals who contribute a reasonable level of support in the creation of a copyrighted work, going beyond those who are traditionally considered capable of independently creating such works. This expanded scope recognizes the collaborative nature of creative processes, acknowledging that individuals involved in providing significant assistance or input should also be acknowledged and granted rights in the resulting work. 118 Nicole MARTINEZ, “Can an AI Machine Hold Copyright Protection Over its Work?” Artrepreneur Art Law Journal (1 June 2017), online: Artrepreneur https://alj.artrepreneur.com/ai-machine-copyright/. 119 Virendra AHUJA, “Artificial Intelligence and Copyright: Issues and Challenges” (11 June 2020) ILI Law Review, online: Social Science Research Network https://ssrn.com/abstract=3864922. 120 Denicola, supra note 75 at 281–2. 121 Ibid. VII. Copyright Protection for AI-Generated Output in the European Union By adopting this approach, the aim is to foster a more inclusive and equitable system that rewards and incentivizes col- lective efforts in the production of copyrighted works.121 VIII. Conclusion The question of copyright protection for AI-generated works and the threshold of origin- ality has sparked significant debate and legal interpretation. The concept of originality holds a crucial role in determining eligibility for copyright protection, with variations across jurisdictions. In the ECJ, originality is established as a fundamental criterion for protecting literary and artistic works under EU law. However, for AI-generated works, the ECJ has clarified that the presence of human creativity and free choices is essential to meet the required level of originality for copyright protection. The controversy sur- rounding authorship in AI-generated works has created confusion globally, with inter- national legal frameworks struggling to address the uncertainty and indecisiveness surrounding AI authorship. Prompt action is necessary to navigate and resolve AI-related concerns, ensuring that legal systems keep up with technological advance- ments. Regrettably, many countries’ copyright laws have overlooked these developments, failing to address the challenges posed by AI-generated works adequately. It underscores the need for comprehensive and forward-thinking approaches to copyright law in response to evolving technological landscapes. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press 23 Asian Journal of International Law Within the current copyright legal framework, the protection of AI-generated works raises several important questions. As courts worldwide grapple with the legal uncertain- ties surrounding authorship in AI-generated works, the issue has garnered attention from organizations such as WIPO, which has actively sought to address the intersection of AI and copyright law. Recognizing the intricate relationship between AI-generated works and human intervention, WIPO has initiated discussions and invited submissions on this crucial matter. However, despite these efforts, a consensus on the complex puzzle presented in this article has yet to be reached. Ongoing discussions and inquiries highlight the need for continued exploration and development of legal approaches to address the challenges posed by AI-generated works within the realm of copyright law. g p y g py g The CJEU, in a series of cases, has firmly established that AI-generated works do not qualify for copyright protection due to the absence or lack of human creativity. A notable example is Cofemel – Sociedade de Vestuário SA v G-Star Raw CV,122 where it was clarified that if the realization of a subject matter is determined by technical considerations, rules, or constraints that leave no room for creative freedom, it cannot be considered sufficiently original to be recognized as a work. VIII. Conclusion From this standpoint, it can be inferred that EU law generally grants protection to works produced with the assistance of AI when there are substantial human contributions, distinguishing them from works that are predominantly AI-generated without significant human intervention. This approach aligns with the requirement for human creativity to be a key element in the creation of copyright-eligible works. Certain national jurisdictions, such as the UK, Ireland, New Zealand, Hong Kong, and India, have provisions in their copyright laws that define the author as the person respon- sible for making the necessary arrangements for the creation of the work. While these provisions were not specifically crafted with Generative AI in mind, they can be relevant in discussions about authorship in AI-generated works. However, this approach falls short when the work is entirely generated by computer algorithms, leaving a lacuna in terms of authorship. By contrast, countries such as the US and Australia have established laws that provide protection, but only when the work is created solely by human beings. These divergent approaches highlight the ongoing challenge of addressing AI-generated works’ unique characteristics and complexities within copyright frameworks. This paper has examined the question of copyright protection for AI-generated works and identified several key findings. First, originality is a crucial criterion for copyright eli- gibility, requiring works to reflect the author’s personality through free and creative choices. While fully automated works may struggle to meet the originality threshold, AI-assisted works with human involvement can qualify for protection. Second, the ECJ has played a significant role in shaping the interpretation of originality, emphasizing the importance of creative freedom and the absence of technical constraints. Third, there is a need for ongoing discussions and potential revisions to address the unique chal- lenges posed by AI-generated works, including issues of attribution, ownership, and the definition of authorship. These findings highlight the need for a balanced and adaptable framework that acknowledges the collaborative nature of AI-generated works and ensures equitable protection for both human creators and technological advancements. Moving forward, it is recommended that policymakers and researchers focus on several key areas to address the challenges and opportunities presented by AI-generated works. While there is a theoretical argument suggesting that updates to copyright laws might better encompass and protect AI-generated works, it is essential to approach this with caution, given the complexities and potential implications for the copyright system as a whole. 122 Case (C-683/17) ECLI:EU:C:2019:721, Cofemel - Sociedade de Vestuário SA v G-Star Raw CV. https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Cite this article: GAFFAR H, ALBARASHDI S (2024). Copyright Protection for AI-Generated Works: Exploring Originality and Ownership in a Digital Landscape. Asian Journal of International Law 1–24. https://doi.org/ 10.1017/S2044251323000735 VIII. Conclusion Further research and evidence are needed to substantiate such significant https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press Hafiz Gaffar and Saleh Albarashdi 24 amendments. This entails clarifying the threshold of originality and considering alterna- tive criteria that account for the unique nature of these works. International collaboration is crucial for establishing consistent standards and guidelines across jurisdictions that fos- ter harmonization, facilitate cross-border recognition, and protect AI-generated works. Ethical considerations should also be at the forefront, with guidelines and frameworks developed to address issues of fairness, bias, and accountability in AI creative processes. Standardized licensing and attribution mechanisms specific to AI-generated works should be developed to facilitate recognition, ownership, and appropriate attribution to human creators and AI systems. Finally, public awareness and education initiatives are essential to increase understanding among creators, users, and the general public about AI-generated works, their implications, and the rights and responsibilities associated with them. By addressing these recommendations, policymakers and researchers can cre- ate a supportive and adaptive framework that encourages innovation, protects rights, and fosters responsible use of AI in creative endeavors. Acknowledgements. The authors would like to thank the reviewers for their comments. Funding statement. None. Competing interests. The authors declare none. Competing interests. The authors declare none. Competing interests. The authors declare none. Assistant Professor at Sultan Qaboos University, college of law Dr. Hafiz Gaffar Ibrahim is Assistant Professor of Civil Law at Sultan Qaboos University in the Sultanate of Oman. He holds a PhD degree in Civil Law and a Master’s degree in Private Law from the College of Law at the University of Al-Neelain, Republic of Sudan. He is a practice- oriented academic with extensive experience in areas of law and technology, property law, Torts & contract law. He has a diversified and extensive experience with govern- mental agencies and legal institutions in UAE, such as the Ministry of justice. Prior to joining Sultan Qaboos University, Hafiz worked with University of Sharjah, Al-Jouf University - Kingdom of Saudi Arabia, AL –Neelain University College of law- Republic of Sudan & as a visiting- Assistant Professor at American University in United Arab Emirates. Research interests of Dr. Hafiz are related to law and technology, Comparative Law & Legal Theory & Islamic law. He is an author of numerous articles and had a number of successful media appearances on the aforementioned topics. Associate Professor of Commercial Law, College of Law, Sultan Qaboos University. He holds a PhD degree in Com mercial Law from University of Exeter and a Master’s degree in Private Law from University of Aberdeen. Email: sinaw814@squ.edu.om. Research interests of Dr. Saleh are related to law and technology, Commercial Law & Maritime Law, Arbitration. He is an author of numerous articles and had a number of successful media appearances on the aforementioned topics. Cite this article: GAFFAR H, ALBARASHDI S (2024). Copyright Protection for AI-Generated Works: Exploring Originality and Ownership in a Digital Landscape. Asian Journal of International Law 1–24. https://doi.org/ 10.1017/S2044251323000735 https://doi.org/10.1017/S2044251323000735 Published online by Cambridge University Press	9,596
W4380089907.txt_1	Open-Science-Pile	Open Science	Various open science	2,023	Slope of the power spectral density flattens at low frequencies (&lt;150 Hz) with healthy aging but also steepens at higher frequency (&gt;200 Hz) in human electroencephalogram	None	Unknown	Unknown	7,398	11,494	Cerebral Cortex Communications, 2023, 4, 1–12 https://doi.org/10.1093/texcom/tgad011 Original Article Srishty Aggarwal 1 , Supratim Ray2, * 1 Department 2 Centre of Physics, Indian Institute of Science, Bengaluru 560012, India, for Neuroscience, Indian Institute of Science, Bengaluru 560012, India *Corresponding author: Supratim Ray, Centre for Neuroscience, Indian Institute of Science, Bengaluru 560012, India. Email: sray@iisc.ac.in The power spectral density (PSD) of the brain signals is characterized by two distinct features: oscillations, which are represented as distinct “bumps,” and broadband aperiodic activity, that reduces in power with increasing frequency and is characterized by the slope of the power falloff. Recent studies have shown a change in the slope of the aperiodic activity with healthy aging and mental disorders. However, these studies analyzed slopes over a limited frequency range (<100 Hz). To test whether the PSD slope is affected over a wider frequency range with aging and mental disorder, we analyzed the slope till 800 Hz in electroencephalogram data recorded from elderly subjects (>49 years) who were healthy (n = 217) or had mild cognitive impairment (MCI; n = 11) or Alzheimer’s Disease (AD; n = 5). Although the slope reduced up to ∼ 150 Hz with healthy aging (as shown previously), surprisingly, at higher frequencies (>200 Hz), it increased with age. These results were observed in all electrodes, for both eyes open and eyes closed conditions, and for different reference schemes. However, slopes were not significantly different in MCI/AD subjects compared with healthy controls. Overall, our results constrain the biophysical mechanisms that are ref lected in the PSD slopes in healthy and pathological aging. Key words: EEG; aperiodic activity; Alzheimer’s disease; alpha oscillation. Introduction Neural signals such as electroencephalography (EEG), magnetoencephalography (MEG), electrocorticography (ECoG), and local field potential (LFP) provide critical insights into the physiological processes underlying key aspects of human cognition and neurodevelopment. These signals often reveal oscillations at different frequencies, which are represented as “bumps” in the power spectral density (PSD), and have been extensively studied as an objective measure for cognitive phenotyping (Klimesch 1999; Schutter and Knyazev, 2012), biomarkers for age (Başar 2013; Murty et al. 2020), as well as neurological disorders (Newson and Thiagarajan 2018; Murty et al. 2021). In addition, the aperiodic background activity in the signals, often known as the “1/f component” or “scale-free activity,” is characterized by the slope or exponent of the PSD (on a log–log scale) in a specified frequency band (He 2014). The PSD slope has garnered interest in recent years, as has been shown to be one of the key features of signal variability (Ribeiro and Castelo-Branco 2022) and has been related to N900 lexical prediction (Dave et al. 2018), working memory (Donoghue et al. 2020b), grammar learning (Cross et al. 2022), sleep changes (Bódizs et al. 2021), anesthesia (Kreuzer et al. 2020), and EEG fingerprinting (Demuru and Fraschini 2020). The slope has been suggested to depend on multiple factors that include self–organized criticality (Mandelbrot 2013), excitation-inhibition balance (Freeman and Zhai 2009; Gao et al. 2017), dendritic response to an input (Voytek and Knight 2015), tissue properties (Bédard et al. 2006a), temporal dynamics of the synaptic processes triggered by Poisson spiking (Milstein et al. 2009), and ionic diffusion processes across the extracellular membrane (Bédard and Destexhe 2009). More recently, it has been linked to the “neural noise” hypothesis ((Voytek et al. 2015); more details in the Discussion section). The slope also depends on the type of signal. For example, slope for ECoG is much steeper than LFP between 20 and 100 Hz but becomes comparable between 200 and 400 Hz (Dubey and Ray 2019). Change in aperiodic activity is suggested to be a plausible biomarker for some neurological and psychiatric diseases like schizophrenia (Molina et al. 2020), attention deficit hyperactivity disorder (ADHD; Robertson et al. 2019; Ostlund et al. 2021), and Fragile X Syndrome (Wilkinson and Nelson 2021), although the aperiodic activity was unchanged in other diseases such as Alzheimer’s disease (AD) (Benwell et al. 2020; Springer et al. 2022). Recently, reduction in the slope with increasing age has been observed across several studies under different task paradigms (Voytek et al. 2015; Dave et al. 2018; Tran et al. 2020) or even in resting-state conditions (Thuwal et al. 2021; Hill et al. 2022; Merkin et al. 2023). However, slope analysis in these studies has been restricted to frequency ranges below 125 Hz, with most of them only up to 50 Hz. Indeed, very few reports have studied the PSD slopes in EEG at frequencies above 150 Hz (Dehghani et al. 2010), Received: May 25, 2023. Editorial decision: May 30, 2023 © The Author(s) 2023. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Slope of the power spectral density f lattens at low frequencies (<150 Hz) with healthy aging but also steepens at higher frequency (>200 Hz) in human electroencephalogram 2 \| Cerebral Cortex Communications, 2023, Vol. 4, No. 2 Materials and methods The details of experimental setup and data collection have been explained in detail in previous studies (Murty et al. 2020, 2021; Kumar et al. 2022). The data are already published; here, we carried out further analysis of the slope of the PSD which was computed over a much wider frequency range than before. In addition, we have used a novel dataset involving “eyes closed” condition, which was not used previously. We summarize the details below in brief. Dataset We used the EEG dataset collected from 257 human subjects (148 males and 109 females) aged 50–88 years (Murty et al. 2020, 2021) under Tata Longitudinal Study of Aging (TLSA) who were recruited from the urban communities of Bengaluru, Karnataka, India. They were clinically diagnosed by psychiatrists, psychologists and neurologists at National Institute of Mental Health and Neurosciences (NIMHANS), and M.S. Ramaiah Hospital, Bengaluru as cognitively healthy (n = 236), or with MCI (n = 15) or AD (n = 6), using a combination of tests such as the Clinical Dementia Rating scale (CDR), Addenbrook’s Cognitive Examination-III (ACE-III), and Hindi Mental State Examination (HMSE). Diagnosis of all MCI/AD subjects was reviewed by a panel of four experts who reclassified two MCI subjects as healthy. Data from these two subjects were not taken for analyses. Further, 11 (10 healthy and 1 MCI) subjects were discarded due to noise (See Artifact Rejection subsection). One AD patient was further discarded as there was no age and gender matched control. We further discarded another 10 subjects (9 healthy and 1 MCI) for whom data was collected using 32 channels, leading to the usable 64-channel data of 233 subjects (217 healthy, 11 MCI and 5 AD). The eyes closed data (see the next subsection) was not collected from 16 subjects (13 healthy, 2 MCI, and 1 AD), leading to a slightly smaller dataset for the eyes closed condition. Informed consent was obtained from the participants of the study and monetary compensation was provided. All the procedures were approved by the Institute Human Ethics Committees of Indian Institute of Science, NIMHANS, and M.S. Ramaiah Hospital, Bengaluru. Experimental settings and behavioral task Brief ly, EEG was recorded from 64-channel active electrodes (actiCap) using BrainAmp DC EEG acquisition system (Brain Products GMbH). The electrodes were placed according to the international 10–10 system, referenced online at FCz. Raw signals were filtered online between 0.016 Hz (first-order filter) and 1 kHz (fifth-order Butterworth filter), sampled at 2.5 kHz, and digitized at a 16-bit resolution (0.1 μV/bit). The subjects were asked to sit in a dark room in front of a gamma corrected LCD monitor (BenQ XL2411; dimensions: 20.92 × 11.77 inches; resolution: 1,280 × 720 pixels; refresh rate: 100 Hz) with their heads supported by a chin rest. It was placed at (mean ± SD) 58 ± 0.7 cm from the subjects (range: 54.9–61.0 cm) and subtended 52◦ × 30◦ of visual field for full screen gratings. Eye position was monitored using EyeLink 1000 (SR Research Ltd), sampled at 500 Hz. In the beginning of the experiment, the resting state EEG for the eyes closed condition was collected for 1–2 min. Then, the subjects performed a passive visual fixation task, including the full screen grating stimuli. The task consisted of a single session that lasted for ∼ 20 min, divided in 2–3 blocks with 3–5 min breaks in between, according to subjects’ comfort. Every trial started with the onset of a fixation spot (0.1◦ ) shown at the centre of the screen, on which they were instructed to fixate. After an initial blank period of 1,000 ms, two to three full screen grating stimuli were presented for 800 ms with an interstimulus interval of 700 ms using a customized software running on MAC OS. The stimuli were full contrast sinusoidal luminance achromatic gratings with either of the three spatial frequencies (one, two, and four cycles per degree (cpd)) and four orientations (0◦ , 45◦ , 90◦ , and 135◦ ). Our analyses were restricted to 500 ms of the interstimulus period before the onset of the stimulus, referred as the baseline period in the previous studies (Murty et al. 2020, 2021). We refer to it as “eyes open” condition. Artifact rejection For eyes open data, we used the artifact rejection framework as described in (Murty et al. 2020, 2021; Murty and Ray 2022) and the steps are summarized here: (a) Eye-blinks or change in eye position outside a 5◦ fixation window during −0.5 to 0.75 s from stimulus onset were noted as fixation breaks and removed off line. This led to a rejection of 14.6 ± 2.8% (mean ± SD) repeats. (b) All the electrodes with impedance > 25 k were rejected. Impedance of final set of electrodes was (mean ± SD) 5.48 ± 1.83 k (c) In the remaining electrodes, outliers were detected as repeats with deviation from the mean signal in (i) time or (ii) frequency domains by more than 6 standard deviations, and subsequently electrodes with more than 30% outliers were discarded. (d) Further, repeats that were deemed bad in the visual electrodes (P3, P1, P2, PO3, POz, PO4, O1, Oz, and O2) or in more than 10% of the other electrodes were considered bad, Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 but the effect of aging or mental disorder on the PSD slopes at these frequencies is unknown. Higher frequencies have generally been ignored because they have low absolute power and therefore could be more susceptible to instrumentation noise or other artifacts related to muscle activity. However, the definitions of “noise” and “signal” are rather nuanced (see (Uddin 2020), for a comprehensive discussion)—“1/f noise” could arise due to signal variability organized across various spatial and temporal scales, and therefore the higher frequencies could ref lect important information about neural processes occurring over fast timescales, such as synaptic neurotransmitter diffusion time (Sabatini and Regehr 1996). For example, power in the “high-gamma” range (>80 Hz) has been shown to accurately ref lect multi-unit firing in the LFP (Ray and Maunsell 2011), with a very high correlation between 250 and 500 Hz. Similarly, the oscillations in high-frequency range (HFR, 300–500 Hz) have been shown to exhibit age-dependent changes (Nakano and Hashimoto 1999, 2000). This prompted us to investigate the aperiodic activity in these HFRs. In this study, we explored aperiodic activity in the EEG activity up to 800 Hz in healthy elderly subjects aged 50–88 years. We analyzed the slopes in both eyes open and eyes closed states. Eyes closed state enabled us to minimize potential electromyography (EMG) artifacts that could be present in the eyes open state. We further examined the dependence of slopes on the reference schemes. Finally, we compared the slopes in subjects with mild cognitive impairment (MCI) and early AD with their age and gender matched healthy controls. Srishty Aggarwal and Supratim Ray The eyes closed EEG data were segmented into non-overlapping 2-s epochs, resulting in a higher resolution of 0.5 Hz. Each segment was then treated as a “stimulus repeat” and subjected to the same artifact rejection pipeline as the eyes open data, with minor changes as described below. We only considered subjects who were deemed good for the eyes open condition. Then, we started with step (b) above, yielding the average impedance of 5.47 ± 1.77 k. We then applied the RMS criteria in place of deviation from mean signal in time domain in (c). Since, we were applying RMS criteria at an initial step on the raw data containing the outliers, we had to increase the lower cut off to 2.5 μV and keep the upper cut off same. We repeated the remaining steps for artifact rejection done for eyes open from (c) (ii)—(e), i.e. detection of outliers from standard deviation in frequency domain to rejection of electrodes with slopes less than 0 as described above. It led to the exclusion of (3.11 ± 4.87%) repeats and (18.86 ± 13.10%) electrodes. No additional subject was rejected when we applied the criteria in (g) for eyes closed data. After discarding all the bad repeats, (293.42 ± 67.22) and (39.35 ± 7.75) repeats were available for eyes open and eyes closed conditions, respectively. EEG data analysis Our primary emphasis was to characterize slope of the aperiodic activity in the PSD as a function of age within the elderly 3 population (>49 years), for which we divided these subjects into two groups: 50–64 (Mid) and > 64 years (Old), as depicted in Table 1. We also compared the slopes in subjects with AD/MCI (termed “cases”) with their healthy, age and gender matched controls. As in our previous studies (Murty et al. 2021; Kumar and Ray 2023), for each case, we averaged the relevant metrics for all age (± 1 year) and gender matched controls to yield a single control data point for each case, yielding 16 (13) pairs for eyes open (eyes closed) analyses (Table 1). All the data analyses were done using custom codes written in MATLAB (MathWorks. Inc; RRID:SCR_001622). Similar to the previous studies (Murty et al. 2020, 2021; Kumar et al. 2022) on this dataset, we chose [−500 0] ms as the eyes open period, yielding a resolution of 2 Hz. The analyses were performed using unipolar reference scheme, unless otherwise specified. Power spectrum was obtained using the multi-taper method with a single taper using the Chronux Toolbox ((Bokil et al. 2010), RRID:SCR_005547) for individual trials and then averaged across the trials for each electrode, which reduces the noise in the spectral estimator (Jarvis and Mitra 2001). Slope analysis The slope of the 1/f aperiodic component of the trial-averaged PSD for each electrode was computed using the Matlab wrapper for the Fitting Oscillations and One Over f (FOOOF) toolbox (Donoghue et al. 2020b). In FOOOF, the power spectrum P(f ) for frequency f is modeled as a combination of aperiodic AP(f) and oscillatory components and can be expressed as P(f ) = AP(f ) + n Gn (f ). The AP(f) is given by AP(f ) = 10b f −χ , where χ is the exponent or “slope” and b is the offset. Each oscillatory contribution Gn (f ) is modeled as a Gaussian peak: f − μn Gn (f ) = an exp − 2σn2 2 , with an as the peak height, μn as the centre frequency and σn as the width of each component. The settings chosen for FOOOF model parameters were: peak width limits: [4,8] for eyes open and [1,8] for eyes closed (different values were used to account for differences in frequency resolution in the two datasets); maximum number of peaks: 5; minimum peak height: 0.2; peak threshold: 2.0; and aperiodic mode: “fixed.” In our data, peaks in the PSD were only evident in the alpha range (8–12 Hz) and line noise (50 Hz) and its harmonics. By taking a frequency range that did not include these oscillatory frequencies (or by not considering these frequencies for analysis), the slope can also be estimated by simply fitting a straight line to log(P(f )) vs log(f ) plot, as done in a previous study (Shirhatti et al. 2016). We employed least-squares minimization using the program fminsearch in Matlab to obtain the slope, which gave similar results to the slopes estimated using FOOOF. This was done in order to verify the results that were obtained using FOOOF and to also validate the FOOOF fitting procedure for higher frequency ranges. Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 eventually yielding a set of common bad repeats for each subject. Overall, this led us to reject (mean ± SD) 15.58 ± 5.48% repeats. (e) We computed slopes for the power spectrum between 56 and 84 Hz for each unipolar electrode and rejected electrodes whose slopes were less than 0. Overall, it led to a rejection of (19.45 ± 14.64%) electrodes. (f) We found that a small fraction of electrodes/stimulus repeats had either very small or very large signals that were not getting discarded using the pipeline above. Therefore, in addition to the artifact detection pipeline that was used in the previous studies, for each electrode, we computed the root mean square value (RMS) of the time series for all the remaining trials (after removing common bad repeats using the above criteria), and declared repeats having RMS values lower than 1.25 μV or higher than 35 μV as new outliers. We then rejected the electrodes that had more than 30% new outliers. Further, any new outlier if it belonged to the visual electrodes or was common in more than 10% of other electrodes was considered a bad repeat and appended to the existing list of bad repeats. This led to rejection of additional 1.24 ± 1.72 electrodes and 0.16 ± 1.05% repeats. This condition was added mainly to improve PSD plots unlike the previous studies (Murty et al. 2020, 2021; Kumar et al. 2022) that dealt mainly with change in power. The main results remained similar even without applying this new criterion. (g) We further discarded the blocks that did not have at least a single good unipolar electrode in the left visual anterolateral (P3, P1, PO3, O1), right visual anterolateral (P2, P4, PO4, O2), and posteromedial (POz, Oz) electrode groups. We then pooled data across all good blocks for every subject separately for the final analysis. Those subjects who did not have any analyzable blocks (10/237 healthy and 1/15 MCI) were discarded for further analysis. \| 4 \| Cerebral Cortex Communications, 2023, Vol. 4, No. 2 Table 1. Number of subjects in each group for the eyes open and eyes closed conditions. The numbers in parenthesis indicate the male (M) and female (F) subjects separately. Eyes open Eyes closed Mid (50–64 years) Old (>64 years) MCI AD 90 (40 M, 50 F) 82 (37 M, 45 F) 127 (79 M, 48 F) 122 (74 M, 48 F) 11 (9 M, 2 F) 9 (8 M, 1 F) 5 (3 M, 2 F) 4 (2 M, 2 F) Linear regression The slope was modeled to vary linearly with age. We used fitlm function in Matlab that generates the model parameters β1, β2 corresponding to y = β1 + β2 ∗ x, R-squared (R2 ) and P-value using t-statistics. Electrode grouping Scalp maps were generated using the topoplot function of EEGLAB toolbox ((Delorme and Makeig 2004), RRID:SCR_007292) with standard Acticap 64 unipolar montage of the channels. We divided the electrodes into five groups as occipital (O1, Oz, O2, PO3, PO4, PO7, PO8, PO9, PO10, POz), centro-parietal (CP1, CP2, CP3, CP4, CP5, CP6, CPz, P1, P2, P3, P4, P5, P6, P7, P8, Pz), fronto-central (FC1, FC2, FC3, FC4, FC5, FC6, C1, C2, C3, C4, C5, C6, Cz), frontal (Fp1, Fp2, F1, F2, F3, F4, F5, F6, F7, F8, Fz, AF3, AF4, AF7, AF8), and temporal (T7, T8, TP7, TP8, TP9, TP10, FT7, FT8, FT9, FT10). FCz and Fpz were used as the reference and the ground, respectively. In our previous studies (Murty et al. 2020, 2021; Kumar et al. 2022), power analysis was done on electrodes for which strong gamma power was observed (P3, P1, P2, PO3, POz, PO4, O1, Oz and O2), which were termed as “high priority” electrodes. We therefore started with the slope analysis on this group and later extended to all the electrode groups. Statistical analysis The statistical analysis was done for slope comparison. Kruskal– Wallis (K-W) test, Wilcoxon Rank Sum (WRS) test were used to compare the medians across groups to be consistent with previous studies (Murty et al. 2020, 2021). The standard error of median (SEM) was computed after bootstrapping over 10,000 iterations. Similar results were obtained when we used the mean instead of the median and ANOVA instead of K-W test (data not shown). To remove false alarms created by low p-values at certain frequencies, we applied cluster correction (Cohen 2014). We used bwconncomp function of Image Processing toolbox in Matlab to identify the frequency clusters having p-values less than 0.05 and 0.01. We called a cluster significant if at least three consecutive frequency bins were significant. For correction for multiple comparisons across electrodes in topoplots, we used the false discovery rate (FDR) using the Benjamini–Hochberg procedure for independent tests (Benjamini and Hochberg 1995). Results We examined EEG data for 217 healthy adults aged between 50 and 88 years, grouped as Mid (50–64 years; n = 90) and Old (>64 years; n = 127). The subjects sat with their eyes closed for 1–2 min before performing a fixation task in which full-screen gratings were presented for 800 ms with an inter-stimulus interval of 700 ms. We computed the PSD of 500 ms segments of data during interstimulus period to get the “eyes open” condition, yielding a frequency resolution of 2 Hz. To overcome the plausible electromyography (EMG) artifacts in eyes open state, we also analyzed the eyes closed data with segments of 2 s each, which yielded a higher resolution of 0.5 Hz. We computed the slopes using Matlab wrapper for FOOOF toolbox (Donoghue et al. 2020b), in steps of 20 Hz between 4 and 1,000 Hz with centre frequencies from 40 to 960 Hz by taking PSD segments of ± 50 Hz around each centre frequency. The first and the last PSD segments were slightly smaller owing to frequency limit of 4–1,000 Hz. Slopes less than 0.01 were not considered for analysis (see Materials and Methods). We have shown the PSDs and slopes up to 800 Hz to avoid the inf luence of filter roll off caused by a fifth order Butterworth filter at 1,000 Hz. Slopes vary differently with age in low- and high-frequency regimes Figure 1(A) shows the median PSDs and slopes with frequencies for the mid and old groups for the eyes open condition for the “high-priority” electrode group (see Methods for details). Like the slope variation in LFP data in monkeys (Shirhatti et al. 2016), for both age groups, slope was high initially due to the presence of oscillatory activities and decreased beyond ∼ 60 Hz until ∼ 140 Hz. It then increased again after a “knee” at ∼ 175 Hz (see bottom plot in Fig. 1A). The PSDs f lattened out for old subjects than mid aged subjects from ∼ 50 Hz until ∼ 140 Hz, resulting in lower slopes for the former in the LFR 64–140 Hz, as highlighted by yellow boxes. This is in consensus with the results reported previously (Voytek et al. 2015; Merkin et al. 2023). However, beyond ∼ 200 Hz, the trend reversed and the PSDs for old subjects became steeper, resulting in steeper slopes beyond ∼ 200 Hz up to ∼ 700 Hz. The HFR at 230–430 Hz, marked in orange, highlights the prominent region of higher slopes for the old group. Since the frequency resolution for the eyes open condition was 2 Hz, and there could be some EMG activity related to maintenance of fixation, we also studied the slopes when eyes were closed for which longer segments of data were used to get a frequency resolution of 0.5 Hz (Fig. 1B). This condition revealed Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Since the slope does not remain constant throughout the full frequency range (Shirhatti et al. 2016), we fitted the PSD segments in frequency ranges from 4 to 1,000 Hz in steps of 20 Hz with the centre frequency between 40 and 960 Hz, having the frequency width of 100 Hz. The PSD segments with the lowest and highest centre frequencies had slightly smaller frequency range width owing to frequency limit of 4–1,000 Hz. We also fitted a single slope in the low-frequency range (LFR) (64–140 Hz) and the HFR (230–430 Hz) to verify that the trend in slope is not attributable to a particular frequency range width and length of PSD segments. Note that FOOOF has a limitation that if the frequency range to be fitted has peaks at its ends, the evaluated slope is inappropriate (Gerster et al. 2022). To overcome this limitation, we declared ± 4 Hz around the peaks corresponding to the line noise at 50 Hz and its harmonics as “noise peaks,” and avoided frequency ranges for which these noise peaks lay near their end points. We also did not use slopes that were less than 0.01, which may be due to poor fitting. Srishty Aggarwal and Supratim Ray \| 5 a more prominent alpha peak as compared with the eyes open condition, and also showed a well-known slowing of the alpha wave with aging (Scally et al. 2018; Merkin et al. 2023), with the centre frequency reducing from 9.72 ± 0.11 in mid to 9.41 ± 0.09 in old-aged group (P = 0.008, WRS test). Importantly, the f lattening of slope in LFR and steepening in HFR with age was also observed in this dataset (Fig. 1B, bottom panel). To better quantify the factors that could lead to the change in PSDs, we first compared the change in PSD between mid and old groups by subtracting the log PSD plots for the two conditions shown in Fig. 1(A) and (B) (and multiplying by 10 to get units of decibels; Fig. 1C). As power is subtracted on a log scale, it is simply the log of the ratio of powers, and therefore, represents the scaling factor that must be applied on the PSD for the mid group to obtain the PSD for the old group. This log ratio was negative (scaling factor < 1) below ∼ 50 Hz, and subsequently became positive with a peak between 100 and 200 Hz. This ratio was smaller for the eyes closed condition in the LFR but comparable in the HFR. We also considered an additive noise hypothesis in which noise was added to the signal obtained from the mid group to get the signal for the old group. If the noise at any frequency is independent of the signal, the power is additive, such that the power of the required noise can be obtained by simply subtracting the PSDs but on a linear (rather than log as done in Fig. 1C) scale (Fig. 1D). This also yielded a shallow “bump” between 50 and 500 Hz peaking at around ∼ 100 Hz (values below ∼ 50 Hz are negative due to a reduction of alpha and beta power with aging). As in Fig. 1(C), noise power was lower in LFR for eyes closed than the eyes open condition, potentially ref lecting some contribution of EMG which is prominent between 40 and 200 Hz (Muthukumaraswamy 2013; McManus et al. 2020). However, this “noise” was comparable in the HFR range for the two conditions. Age variation of slope in LFR and HFR is observed in all electrodes Next, we compared the slopes for different sensors, for which the electrodes were grouped in five categories as mentioned in Materials and Methods. Figure 2(A) shows the PSDs and slopes in all the electrode groups for the eyes open condition. Interestingly, the PSD slopes varied considerably depending on electrode location over the entire frequency range. Figure 2(C) shows the fitted slopes in LFR and HFR. In particular, in HFR, the slopes increased radially from the central area. In spite of the differences in absolute slope values, the difference in slope between old and mid aged groups showed consistent trends (Fig. 2B). The difference in slopes between old and mid groups in LFR (Fig. 2C; top plot) and HFR (Fig. 2C; bottom plot) were consistently negative and positive, respectively. So, the posterior– anterior shift in aging (PASA), commonly observed in functional neuroimaging studies of aging and characterized by age-related reduction in occipital activity alongside increase in frontal activity (Mccarthy et al. 2014), was not observed in aperiodic activity in the present dataset. The electrodes for which the differences were significant (P < 0.05, WRS test, false discovery rate (FDR) corrected; red dots in Fig. 2C) were more concentrated in the posterior and occipital electrodes. Figure 2(D) shows the median value of the slopes in two age groups in LFR (top) and HFR (bottom), which were significantly different in all electrode groups (p-values are shown in the plots). The results were similar when the analysis was restricted to only males or females (data not shown). Similar analyses were performed for eyes closed state as shown in Fig. 3. The results in eyes closed state were similar to the eyes open state, although the differences in slopes between the age groups (Fig. 3C) were less prominent and only significant in few electrodes in the occipital area for HFR (Figs. 3C and D). The median goodness of fit (R2 ) for all electrodes across the frequency range in these two conditions is shown in Supplementary Fig. 1. R2 was relatively high at frequencies in LFR and gradually decreased at higher frequencies, potentially due to lower absolute power. R2 was also lower for the eyeclosed condition as compared with eyes open, due to lower number of trials that were used to obtain the PSD in the former case. Interestingly, in LFR, it was significantly higher for midaged group as compared with old-aged group. To rule out the Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Fig. 1. (A) PSDs (top) and slopes (bottom) for the two age groups in high priority electrodes in eyes open state. Solid traces represent the median for mid and old age groups respectively and shaded region around them indicates ± SEM across subjects, computed after bootstrapping over 10,000 iterations. The numbers in legend in the top panel represent the subjects in the respective age groups. Horizontal bars at the abscissa in the bottom panel represent significance of differences of slopes between mid and old (black: P < 0.05 and green: P < 0.01, K-W test, cluster corrected (CC)). Yellow and orange boxes represent the LFR (64–140 Hz) and the HFR (230–430 Hz), respectively. (B) Same as (A) in the eyes closed condition. (C) The median change in power between mid and old in decibels (dB) for the eyes open (purple) and eyes closed (gray) conditions. (D) Median change in absolute power between mid and old for these two conditions. 6 \| Cerebral Cortex Communications, 2023, Vol. 4, No. 2 Fig. 3. Variation of PSDs and slopes with frequency across the two age groups in various areas of brain for the eyes closed state. Same as Fig. 2 but for the eyes closed condition. possibility of differences in R2 affecting our results, we subsampled from each population to match the distribution of R2 values and still found that the slopes remained different between mid and old groups. Further, in HFR, R2 values were not significantly different between the two age groups, providing additional confidence in the slope results in this frequency range. While reporting aperiodic activity, previous studies have included the variation of exponent as well as offset. In general, the rotation of PSD changes the exponent and offset together leading to strong correlation between these two parameters, as reported previously (Hill et al. 2022; Merkin et al. 2023). Changes in offset (while keeping the exponent same) can be observed in task-related studies that corresponds to broadband change in PSD (Voytek and Knight 2015). However, we observed similar changes in offset with age in LFR and HFR as the exponent (Supplementary Fig. 2), and, hence, have focused only on the exponent variation. Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Fig. 2. Variation of PSDs and slopes with frequency across the two age groups for different electrodes for the eyes open state. (A) PSDs and slopes across different electrode groups for mid and old age groups. The top plot in the left bottom panel highlights the electrodes chosen for each electrode group. (B) PSDs and slopes between the age groups for each electrode group separately. Solid traces represent the median and shaded region around them indicates ± SEM across subjects, computed after bootstrapping over 10,000 iterations. The numbers in legend in the top panel represent the subjects in the respective age groups. Colored bars at the abscissa in the bottom panel represent significance of differences of slopes between mid and old (black: P < 0.05 and green: P < 0.01, K-W test (CC)). Yellow and orange boxes represent the LFR (64–140 Hz) and the HFR (230–430 Hz), respectively, in (A) and (B). (C) Topoplot of the difference in slopes between old and mid age groups (left column), computed in LFR (top row) and HFR (bottom row), and the raw slope values in the mid (central column) and old (right column) age groups. Electrodes with significant difference (P < 0.05, WRS test, FDR corrected) in slopes are highlighted in red in the left panel. (D) Bar plots showing median slopes for each electrode group for mid- and old-aged people for LFR and HFR, with error bars representing standard error of median (obtained using bootstrapping). The WRS test p-values are indicated at the top of the bar plots and the dots represent the slope for each subject. Srishty Aggarwal and Supratim Ray \| 7 Slope variation across electrodes is independent of the reference scheme In HFR, the slopes were lower for the central electrodes and increased across frontal, temporal and occipital areas (Fig. 2(C) and Fig. 3(C); bottom panels). In our recording setup, the reference and ground electrodes were near the centre and, therefore, the radial increase in slopes in HFR could be due to an increase with the distance of the electrode from the reference electrode. To test the dependence of HFR slope on the reference scheme, we rereferenced all signals using a bipolar referencing scheme in which each electrode was referenced with respect to a neighboring electrode to yield a virtual bipolar electrode at the mid-point of the two electrodes, yielding 112 bipolar-referenced electrodes (for more details, see Murty et al. 2020). Figure 4 shows the same results as Fig. 2(C) after computing the slopes for the bipolar signals. The results after bipolar referencing remained similar, although less significant, thus repudiating the variation across electrodes as an artifact, although the absolute slopes for bipolar reference scheme were slightly lower than unipolar reference scheme, as also shown previously (Shirhatti et al. 2016). Slopes do not vary between AD/MCI patients and their controls We had 107 healthy subjects for the eyes open and 100 for the eyes closed conditions who were age and gender matched with the AD/MCI subjects (6–19 controls per case subject; see Murty et al. 2021, for details). To make the comparison more balanced, similar to our previous studies (Murty et al. 2021; Kumar and Ray 2023), we computed the median PSDs/slopes across all controls for a given case subject to match the number of cases and controls and subsequently performed pairwise analysis. Figure 5 shows the results for the eyes closed condition (n = 13). The PSDs revealed a significant reduction in beta power (15–35 Hz) in cases compared with controls in all the electrode groups as reported in other studies (Ranasinghe et al. 2022). For example, it reduced from 12.14 ± 1.56 and 11.18 ± 1.32 μV2 in controls to 5.83 ± 1.16 and 5.78 ± 1.24 μV2 in cases in occipital and temporal electrodes (P = 0.0048 and 0.0066, WRS test), respectively. This reduction was observed in the eyes open condition also (n = 16), although it did not reach significance (data not shown). However, there was no significant change in slopes between controls and cases in either LFR or HFR in either eyes open and eyes closed conditions (eyes closed: Fig. 5B–D), consistent with previous studies which also did not find any changes in the PSD slope with MCI/AD (Benwell et al. 2020; Springer et al. 2022). Variation in slope with age is confirmed using regression analysis To confirm the variation in slope across subjects without grouping them in predefined groups, we regressed the slopes in the high priority electrode group with age (see Material and Methods for details) for the two frequency ranges and the two conditions, as shown in Fig. 6. Consistent with previous results, we observed a negative relationship in LFR (slope = −0.010 for eyes open and − 0.013 for eyes closed) and a positive relationship in HFR (slope = 0.017 for eyes open and 0.016 for eyes closed). Interestingly, slope magnitude in HFR was higher than LFR, indicating a faster change of slope with age in HFR. Also, R2 was higher in HFR (0.53 eyes open, 0.46 eyes closed) than LFR (0.36 eyes open, 0.30 eyes closed). All the changes were significant (LFR: P = 0.0007 eyes open, P = 0.0001 eyes closed; HFR: P = 0.0002 eyes open, P = 0.0001 eyes closed). There was nearly equal distribution of AD/MCI cases, shown as pink dots, around the regression line indicating the indifference of their slopes with healthy subjects in both LFR and HFR for the eyes open as well as eyes closed conditions. We used this simple model containing the linear regression analysis to highlight the age-related decrease and increase in slopes in LFR and HFR respectively. However, as shown in Fig. 6, the inter-subject data were noisy and the linear model could be too simplistic, as the characteristics of the slope of the ongoing Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Fig. 4. Same as Fig. 2(C) but for the bipolar reference scheme. 8 \| Cerebral Cortex Communications, 2023, Vol. 4, No. 2 Fig. 6. Scatter plots showing the slopes in high priority electrodes for all subjects in LFR and HFR for baseline and eyes closed conditions. Blue dots represent healthy subjects while MCI and AD subjects are indicated using pink dots. The regression line is in black and the corresponding parameters obtained using regression analysis are shown in top right side of each subplot. activity in brain are dynamic. One possible mechanism for this dynamic change is a phenomenon called self-organized criticality (Bak et al. 1987), which may give rise to nonlinearity in the association of age with slope of the 1/f component. Indeed, origin of “1/f noise” may lie in tissue properties (Bédard et al. 2006a), which could change in a non-linear fashion with age. Nevertheless, the linear model is sufficient to validate the observed pattern in agerelated changes in slopes, as shown by age-wise grouping the subjects in previous figures. Discussion We investigated age and MCI/AD-related changes in the EEG aperiodic activity by studying the variation in the slope of the PSD over a broad frequency range (up to 800 Hz). In a task where subjects maintained fixation on the screen while visual stimuli were presented (Murty et al. 2020, 2021), we computed the PSDs during the inter-stimulus interval period. Consistent with previous studies, PSD slope f lattened with age up to ∼ 150 Hz. However, the slope showed a surprising increase with age at frequencies beyond 200 Hz. This age-related distinct behavior of slopes at LFRs and HFRs was observed in all sensors, although, the slopes varied with the sensor location. These results were confirmed using another dataset from the same subjects when they sat quietly with their eyes closed, for which PSDs could be obtained at higher frequency resolution. This ruled out the possibility of potential EMG artifacts related to open eyes or poor frequency resolution in the eyes open data affecting our results. The results remained similar when we Downloaded from https://academic.oup.com/cercorcomms/article/4/2/tgad011/7190840 by guest on 18 May 2024 Fig. 5. Comparison of PSDs and slopes with frequency between controls and cases (AD/MCI) for the eyes closed state. (A)–(C) are similar to Figs. 2(A–C). Since there were several controls per case, median PSDs/slopes across all controls for each case subject was used to match the numbers of cases and controls. (D) The bar plots show the median slope for each electrode group in LFR and HFR across controls and cases. Individual case and control pair are shown as connected white dots. The WRS test p-values are indicated on the top of the bar plots. Srishty Aggarwal and Supratim Ray used a bipolar reference scheme instead of the original unipolar reference scheme. Further, although we observed a reduction in beta power in subjects with MCI or AD compared with their age and gender matched healthy controls, we did not find any significant change in slopes. Previous studies on PSD slope changes with age Neural mechanisms underlying PSD slope changes Although precise generative mechanisms underlying aperiodic activity is unknown, a multitude of factors have been suggested to describe the slopes, which include tissue properties (Bédard et al. 2006a), self-organized criticality (SOC; He et al. 2010; Chaudhuri et al. 2018), filtering properties of dendrites and extracellular medium (Bédard et al. 2006b; Logothetis et al. 2007; Bédard and Destexhe 2009), and excitation-inhibition (E/I) balance (Gao et al. 2017; Naskar et al. 2021). In addition, the PSD slopes depend on simple factors such as the reference scheme used for recording the data (Shirhatti et al. 2016). Recently, a “neural noise” hypothesis has been proposed to explain the reduction in the PSD slope, which is related with increased asynchronous background neuronal firing (Voytek et al. 2015) or an increase in the E/I ratio (Gao et al. 2017). Specifically, Gao and colleagues developed a model in which Poisson distributed spikes arriving at excitatory and inhibitory neurons generated a change in excitatory and inhibitory conductance that were modeled as a difference-of-exponentials due to rise and decay time constants associated with AMPA and GABAA receptors. Both these conductance profiles produced a low-pass effect, but slower kinetics of GABAA caused a steeper roll-off with frequency compared with the AMPA conductance (see fig. 1 of Gao et al. 2017). Consequently, reduction in inhibition (increase in the E/I ratio), potentially due to dysfunctional inhibitory circuitry with aging, led to shallower slopes. This hypothesis has been supported by experiments in which the PSD slope was modulated by administration of drugs that led to either increased inhibition (e.g. propofol) or increased excitation (ketamine) (Gao et al. 2017; Lendner et al. 2020; Waschke et al. 2021). Our results could be consistent with this hypothesis, provided we modify the characteristics of this neural noise, which was band-pass as shown in Fig. 1(D). 9 Interestingly, surface EMG signals have energy between 40 and 200 Hz, peaking around ∼ 100 Hz, which could explain at least part of the slope changes in the LFR (Muthukumaraswamy 2013; McManus et al. 2020). On the other hand, this cannot explain the effect in steepening of the slope in HFR, since the noise was similar whether eyes were open or closed and occurred at a higher frequency range than EMG activity. Indeed, the noise need not be myogenic but instead generated by the brain. As the timescales of fundamental neural processes like synaptic neurotransmitter diffusion time and timing of spike propagation lie below 10 ms (Sabatini and Regehr 1996; Shepherd 2004), highfrequency activity (>100 Hz) contains a wealth of information about neural processing and integration. Hence, the increase in slope in old group in HFR could be a ref lection of agerelated changes in these neural processes. In addition, it could be due to age-related alteration of physiological properties in brain tissue (Aalami et al. 2003; Lee and Kim 2022), cortical thickness (Lillie et al. 2016), and skin conductance (Lim et al. 1996; Venables and Mitchell 1996), which could determine the amplitude of this high-frequency signal measured from the scalp.	26,007
https://openalex.org/W2058786081	OpenAlex	Open Science	CC-By	2,012	Association of Eleven Common, Low-Penetrance Colorectal Cancer Susceptibility Genetic Variants at Six Risk Loci with Clinical Outcome	Janelle M. Hoskins	English	Spoken	6,752	12,172	Abstract Background: Low-penetrance genetic variants have been increasingly recognized to influence the risk of tumor development. Risk variants for colorectal cancer (CRC) have been mapped to chromosome positions 8q23.3, 8q24, 9p24.1, 10p14, 11q23, 14q22.2, 15q13, 16q22.1, 18q21, 19q13.1 and 20p12.3. In particular, the 8q24 single nucleotide polymorphism (SNP), rs6983267, has reproducibly been associated with the risk of developing CRC. As the CRC risk SNPs may also influence disease outcome, thus in this study, we evaluated whether they influence patient survival. Methodology/Principal Findings: DNA samples from 583 CRC patients enrolled in the prospective, North Carolina Cancer Care Outcomes Research and Surveillance Consortium Study (NC CanCORS) were genotyped for 11 CRC susceptibility SNPs at 6 CRC risk loci. Relationships between genotypes and patient survival were examined using Cox regression analysis. In multivariate analysis, patients homozygous for the CRC risk allele of rs7013278 or rs7014346 (both at 8 q24) were only nominally significant for poorer overall survival compared to patients homozygous for the protective allele (hazard ratio = 2.20 and 1.96, respectively; P,0.05). None of these associations, however, remained statistically significant after correction for multiple testing. The other nine susceptibility SNPs tested were not significantly associated with survival. Conclusions/Significance: We did not find evidence of association of CRC risk variants with patient survival. nclusions/Significance: We did not find evidence of association of CRC risk variants with patient surviva Citation: Hoskins JM, Ong P-S, Keku TO, Galanko JA, Martin CF, et al. (2012) Association of Eleven Common, Low-Penetrance Colorectal Cancer Susceptibility Genetic Variants at Six Risk Loci with Clinical Outcome. PLoS ONE 7(7): e41954. doi:10.1371/journal.pone.0041954 Editor: Amanda Ewart Toland, Ohio State University Medical Center, United States of America Editor: Amanda Ewart Toland, Ohio State University Medical Center, United States of America Received January 31, 2012; Accepted June 29, 2012; Published July 27, 2012 kins et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits tion, and reproduction in any medium, provided the original author and source are credited. Copyright: 2012 Hoskins et al. This is an open-access article distributed under the terms of the Creative Commons Attribut unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by theNational Institutes of Health (NIH) Pharmacogenetics Research Network (U01 GM63340). Abstract The NC CanCORS and GI SPORE were supported by NIH U01 CA 093326 and P50 CA 106991 respectively. The study also received support from the Center for Gastrointestinal Biology and Disease (P30 DK034987). PO was supported by the Department of Pharmacy, National University of Singapore. No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. * E-mail: phaops@nus.edu.sg . These authors contributed equally to this work. ¤a Current address: Department of Pharmacy, Faculty of Science, National University of Singapore, Singapore, Singapore. ¤b Current address: Department of Environmental Health Sciences, Tulane University School of Public Health and Tropical Medicine, New Orleans, Louisiana, United States of America. ¤a Current address: Department of Pharmacy, Faculty of Science, National University of Singapore, Singapore, Singapore. ¤b Current address: Department of Environmental Health Sciences, Tulane University School of Public Health and Tropical Medicine, New Orleans, Louisiana, United States of America. [2–5]. For germline mutations, high-penetrance changes in adenomatosis polyposis coli, mismatch repair, mothers against decapentaplegic homolog 4, bone morphogenetic protein receptor type IA and serine threonine kinase 11 genes have been reported to be associated with increased CRC susceptibility in 5% of the population [6] while the effects of combinations of low-penetrance variants remains largely elusive. Janelle M. Hoskins1., Pei-Shi Ong1,.¤a, Temitope O. Keku2, Joseph A. Galanko2,3, Christopher F. Martin2, Clint A. Coleman1¤b, Michelle Wolfe2, Robert S. Sandler2, Howard L. McLeod1,4,5 1 Division of Pharmacotherapy and Experimental Therapeutics, Institute for Pharmacogenomics and Individualized Therapy, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America, 2 Division of Gastroenterology and Hepatology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America, 3 Center for Gastrointestinal Biology and Disease, Division of Gastroenterology and Hepatology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America, 4 Division of Hematology and Oncology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America, 5 The Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America PLoS ONE \| www.plosone.org Association of Eleven Common, Low-Penetrance Colorectal Cancer Susceptibility Genetic Variants at Six Risk Loci with Clinical Outcome Janelle M. Hoskins1., Pei-Shi Ong1,.¤a, Temitope O. Keku2, Joseph A. Galanko2,3, Christopher F. Martin2, Clint A. Coleman1¤b, Michelle Wolfe2, Robert S. Sandler2, Howard L. McLeod1,4,5 DNA Extraction and SNP Genotyping yp g Buffy coat was prepared from a blood sample collected from each study participant. DNA was extracted from buffy coat using Puregene kit (Qiagen, Valencia, CA, USA). In addition, DNA samples comprising of 94 European-Americans, 93 Han Chinese (Han people of Los Angeles) and 94 Mexican-Americans (Mexican-American community of Los Angeles) from Coriell (Coriell Institute for Medical Research, Camden, NJ, USA) as well as 86 African-Americans as previously described [24], were also used to confirm the reliability and reproducibility of the genotyping performed. All DNA samples were genotyped for 11 CRC susceptibility SNPs at 6 CRC risk loci using TaqMan allelic discrimination assays (Applied Biosystems, Foster City, CA, USA) (Table S1). In a total volume of 20 mL, reactions consisted of 2 mL of 10 ng/mL DNA, 0.5 mL of 20X TaqManH SNP genotyping assays (Applied Biosystems), 10 mL of 2X TaqManH Universal PCR Master Mix, No AmpErase UNG (Applied Biosystems) and 7.5 mL of water. PCR was performed with an initial denaturation step at 95uC for 10 min followed by 50 cycles of denaturation at 92uC for 15 s and annealing with extension at 60uC for 1 min. All PCR reactions were performed on a Bio-Rad Tetrad 2 Thermal Cycler (Bio-Rad, Hercules, CA, USA). The fluorescence intensities of the samples were measured before and after PCR using an Applied Biosystems 7500 Real-Time PCR System (Applied Biosystems). Data obtained was analyzed and genotypes assigned using 7300 System SDS Software, version 1.4 (Applied Biosys- tems). Genotype allocation was performed blinded to patients’ clinical data. Genotyping was successful in .96% of samples. For the samples from the four additional populations, all genotypes were in agreement with that reported in the Hapmap (www. hapmap.org). No discordance in genotype or allele frequencies was observed between the European-American or African-American patient samples with the respective European-Americans or African-Americans in the additional populations genotyped. Besides the SNPs on 8q24, Tomlinson et al identified rs16892766 on 8q23.3 as an additional risk variant [15]. Likewise, Zanke and colleagues found an association between rs719725 on 9p24 and CRC [17]. Subsequent investigation of this site by Poynter et al showed that individuals with the A/A genotype had a moderate increased risk of CRC compared to C/C individuals in population-based but not clinic-based families [13]. On the 10p14 and 11q23 loci, rs10795668 and rs3802842 were respectively shown to be related to disease occurrence [14,15]. Statistical Analysis All polymorphisms were examined for deviation from Hardy- Weinberg equilibrium (HWE) using x2 test. Clinical and biological variables were examined for associations with individual SNPs using Fisher’s Exact Test. Univariate and multivariate survival analyses were carried out using Cox regression analysis to evaluate associations between genetic variants and overall survival. Multivariate models were adjusted for age, ethnicity, gender, disease stage, site of tumor and treatment administered. For each SNP, the risk allele was defined as the allele previously established in the literature to confer a risk of CRC while the other allele was considered the protective allele. Homozygosity for the protective allele served as a reference genotype for regression analysis and was assigned a hazard ratio (HR) of 1.0. A P,0.05 was considered significant. Multiple correction was performed using the conser- vative Bonferroni method. All data was analyzed using SAS statistical analysis software version 9 (SAS Institute Inc, Cary, NC, USA). DNA Extraction and SNP Genotyping In addition, numerous other risk loci have been subsequently identified and mapped to 14q22.2 (rs4444235, BMP4) [18], 15q13 (rs4779584, rs10318, CRAC1) [11], 16q22.1 (rs9929218, CDH1) [18], 18q21 (rs4939827, rs12953717, rs4464148, SMAD7) [8], 19q13.1 (rs10411210, RPHN2) [18] and 20p112.3 (rs961253) [18]. Despite the increasing number of loci being identified to influence the risk of CRC development, to date, only a few studies have investigated the effects of these variants on disease outcome [19–22]. The findings of these studies however remained inconsistent on the relationship between these risk variants and CRC survival. Therefore, in the present study, we examined the prognostic significance of 11 CRC susceptibility SNPs at 6 CRC risk loci (rs6983267, rs10505477, rs7013278, rs7014346, rs719725, rs10795668, rs3802842, rs4779584, rs10318, rs4464148 and rs4939827, Table S1), using 583 patients with CRC from the prospective North Carolina (NC) Cancer Care Outcomes Research and Surveillance Consortium Study (Can- CORS). CRC Risk Variants Don’t Influence Clinical Outcome suggests that a number of allelic variants found in more than 1–5% of the population genetically influences the susceptibility to common heritable diseases [7]. In line with this model, candidate gene analysis and multi-stage genome wide association studies (GWAS) have identified numerous single nucleotide polymor- phisms (SNPs) across several chromosomes that are associated with an increased risk of CRC development [8–18]. In a case-control study involving 1807 patients and 5511 controls, Haiman and colleagues noted that rs6983267 on chromosome 8q24, a genomic region that contains few genes, was significantly linked to a higher predisposition of CRC in individuals of different ethnicities [10]. Several later reports confirmed rs6983267 as a low-penetrance risk marker for CRC [9,12,13,16,17]. Given this consistent observa- tion and the frequent amplification of this region in CRC [10], further analysis of SNPs within this gene-poor area revealed a second tightly linked variant, rs10505477, as well as three other SNPs on 8q24, rs10808556, rs7014346 and rs7013278, as low- penetrance variants that also influence carcinoma formation [10,13,14,15,17]. further confirmed through ascertainment of death records through the social security death index (SSDI) using patients’ social security numbers. Mortality information or survival status was available for all patients genotyped in this study through the SSDI. Disease-free survival information was not available. This study was approved by the Institutional Review Board of the University of NC (IRB number: 04–08–60). Written informed consent was obtained from each participant. Introduction Colorectal cancer (CRC) is the second most common cause of cancer-related death in the United States. Despite improvements in treatment modalities, the 5-year survival rate for CRC patients ranges from 10–90% [1]. This huge variation in clinical outcome is due, in part, to the fact that CRC is a heterogeneous disease comprising discrete subsets that evolve through multiple different etiologies. Both germline and somatic genetic alterations can be involved in the malignant transformation of normal colon cells. Extensive investigations have identified somatic mutations in TP53 or KRAS that are involved in the progression of adenoma to CRC The completion of the Human Genome Project and the development of improved high-throughput genotyping techniques permit large scale interrogation of the genome, resulting in a better understanding of common polygenic disease. This progress has led to the common disease, common variant hypothesis, which PLoS ONE \| www.plosone.org July 2012 \| Volume 7 \| Issue 7 \| e41954 1 July 2012 \| Volume 7 \| Issue 7 \| e41954 CRC Risk Variants Don’t Influence Clinical Outcome CRC Risk Variants Don’t Influence Clinical Outcome Table 1. Demographic, Histopathological and Clinical Characteristics of Study Patients. Patients Characteristics N (%) Patients 583 Age, years Median (range) 65.0 (26.0–93.0) Gender Male 304 (52.1) Female 279 (47.9) Ethnicity European-American 472 (81.0) African-American 111 (19.0) Stage 1 161 (27.6) 2 160 (27.4) 3 165 (28.3) 4 68 (11.7) Missing 29 (5.0) Tumor Site Colon 261 (44.8) Rectum 241 (41.3) Missing 81 (13.9) Treatment - Surgery Yes 573 (98.3) No 8 (1.4) Missing 2 (0.3) Treatment – Chemotherapy Yes 297 (50.9) No 279 (47.9) Missing 7 (1.2) doi:10.1371/journal.pone.0041954.t001 administration (Table S2). For age of CRC diagnosis, apparent differences in genotype distribution were only found for rs10795668 (P = 0.005; Table S2). It was observed that the prevalence of the A/A genotype for this SNP decreases in patients older than 50 years. In addition, stage of disease was significantly related to rs4464148 and rs4939827 genotypes (P,0.05; Table S2). C and T alleles, the risk allele for rs4464148 and rs4939827 respectively, were shown previously to be associated with an increased risk for CRC [8,21]. Surprisingly in the present study, more patients homozygous for the risk allele for both SNPs presented with earlier stage cancer (stages 1–2) at the time of diagnosis, suggesting that these alleles are protective (Table S2). Relationships Between Low-Penetrance CRC Susceptibility SNPs and Overall Survival To test whether the germline variants underlie differences in overall survival in CRC patients, we performed both univariate and multivariate survival analyses. In univariate analysis, a significant difference in survival was only observed in patients with the A/C genotype compared to the A/A genotype for rs3802842 (P,0.05; Table 2). This difference was not significant in a multivariate model (P.0.05; Table 2). In multivariate analysis, patients carrying two risk alleles (T/T) for rs7013278 had reduced survival compared with patients homozygous for protective allele (C/C) (HR = 2.20; 95% CI = 1.24–3.91; P = 0.01; Table 2). Likewise, patients homozygous for the CRC risk allele of rs7014346, A/A, showed inferior overall survival to G/G patients (HR = 1.96; 95% CI = 1.08–3.52; P = 0.03; Table 2). The mod- erate level of significance for these two SNPs was not maintained upon correction for multiple testing. No additional significant association was observed between the rest of the SNPs studied and survival (Table 2). Results The present study was performed to examine the prognostic significance of 11 common, low-penetrance genetic variants at 6 CRC loci that have been previously reported to predispose individuals to CRC [8–17]. Although we found marginal significance between two SNPs, rs7013278 and rs7014346 (HR = 2.20, P = 0.01 and HR = 1.96, P = 0.03 respectively), with inferior CRC survival by multivariate regression analysis, none of these variants showed study-wide association with survival after correction for multiple testing. It therefore highlighted that these known CRC risk variants, do not play a role in influencing CRC mortality, which is in agreement with the findings from three earlier studies [19–21]. Patient Characteristics Table 1 summarizes the baseline characteristics of patients in this study. The median age of patients was 65.0 years old and 47.9% were female. Among the 583 patients, 81.0% were European-Americans and 19.0% were African-Americans. At the time of diagnosis, the proportions of study participants classified to AJCC stages 1, 2, 3 and 4 were 27.6%, 27.4%, 28.3% and 11.7% respectively. In this cohort, the proportion of patients with colon cancer or rectal cancer was comparable. The majority of patients (98.3%) received surgical treatment while chemotherapy was administered to 50.9% of the study population. So far, several prior studies have investigated the association between CRC susceptibility variants with disease progression and survival. Gruber and colleagues were the first to report no correlation between rs10505477 with CRC survival in a predom- inantly Jewish population [19]. In this study, a HR of 1.09 (CI = 0.89–1.32) was found between carriers of risk allele versus non carriers [19] Subsequently, Cicek and colleagues evaluated the influence of 5 other low- penetrance CRC risk markers (rs6983267, rs13254738, rs16901979, rs1447295, DG8S737) on the survival of 460 cases of stages II and III, Caucasian, CRC patients who were the participants of a phase III adjuvant therapy study [20]. While they observed a trend of decreased survival rate with the presence of the rare risk variants (HR for rs6983267 = 1.00, CI = 0.79–1.27; HR for rs13254738 = 1.14, CI = 0.91–1.43; HR for rs16901979 = 1.34, CI = 0.86–2.09; HR for rs1447295 = 1.11, CI = 0.79–1.55; HR for DG8S737 = 1.57, CI = 0.85–2.90) using a log additive model, none of the Study Population and Follow-Up Information Study Population and Follow-Up Information The study design of CanCORS has been described previously [23]. NC CanCORS assembled a prospective population-based cohort in 33 county areas of NC, USA. In this study, DNA samples from 583 eligible patients diagnosed with CRC between April 2003 and January 2005 were retrospectively genotyped. Patient demographics were obtained from a baseline patient survey. Detailed clinical information on primary site of tumor, American Joint Committee on Cancer (AJCC) stage and type of treatment provided were obtained from review of medical records and pathological reports of CRC diagnosis from the North Carolina Central Cancer Registry by trained abstractors within 6 months following diagnosis. Patients were followed up for a median of 3.5 years. Survival and mortality data was determined from a three-year phone survey of participants or next-of kin and PLoS ONE \| www.plosone.org July 2012 \| Volume 7 \| Issue 7 \| e41954 2 CRC Risk Variants Don’t Influence Clinical Outcome PLoS ONE \| www.plosone.org Low-Penetrance CRC Susceptibility Genotypes The distribution of genotypes in this patient cohort is presented in Table S2. For this group of patients, no deviation from HWE was detected among European- or African-American patients for any of the SNPs. The allele frequencies for all SNPs were similar to frequencies of European- (CEU) and African-American populations (ASW) reported in HapMap (www.hapmap.org). Genotype distri- butions varied between European- and African-American patients for all SNPs (P,0.05) except rs7014346 and rs3802842 (Table S2). Relationships Between Low-Penetrance CRC Susceptibility SNPs and Clinicopathological Features of CRC Patients CRC Risk Variants Don’t Influence Clinical Outcome For rs4779584, the result of this study (HR = 1.01, CI = 0.46–2.22) and that of Tenesa et al (HR = 0.95, CI = 0.85–1.06) [21] showed no significant influence of the risk allele of this SNP with CRC survival. This deviates from the findings of Xing et al [22]. A plausible explanation for this deviation is a much higher frequency of the risk allele among the Han Chinese compared to the population in this study and that of Tenesa and colleagues [21], which were predominately of European ancestry. While the allele frequencies of rs4779584 in the Han Chinese CRC patients were not directly reported in the paper by Xing et al [22], a calculation of the allele frequencies of this population based on the genotype of patients presented showed that the calculated allele frequencies (C = 0.19 and T = 0.81) were congruent with that from the Chinese populations in the Hapmap (CHB: C = 0.18 and T = 0.82; CHD: C = 0.16 and T = 0.84) and the Han Chinese population that we have previously genotyped (C = 0.17 and T = 0.83) as part of the four additional populations used for genotyping control. Based on the data from Hapmap and our data of the four additional populations genotyped (data not shown), the allele frequency for the risk allele (T) of rs4779584 was highest in the Han Chinese, followed by African-American and lastly European-American. For rs10795668, HR for overall survival in our study and that by Tenesa et al [21] were 0.82 (CI = 0.37–1.83) Table 2. Associations between genotypes and overall survival of study patients. Susceptibility SNPs and Clinicopathological Features of CRC Patients No significant differences were observed between the SNPs with gender, tumor site, surgical intervention or chemotherapy PLoS ONE \| www.plosone.org 3 July 2012 \| Volume 7 \| Issue 7 \| e41954 July 2012 \| Volume 7 \| Issue 7 \| e41954 CRC Risk Variants Don’t Influence Clinical Outcome CRC Risk Variants Don’t Influence Clinical Outcome Univariate Multivariate# SNP and Genotype Risk Allele Hazard Ratio 95% CI P Hazard Ratio 95% CI P rs6983267 T/T G 1.00 – – 1.00 – – G/T 0.85 0.51–1.41 0.53 1.07 0.59–1.93 0.83 G/G 1.17 0.71–1.92 0.54 1.35 0.73–2.51 0.35 rs10505477 G/G A 1.00 – – 1.00 – – A/G 0.90 0.55–1.48 0.68 1.14 0.64–2.06 0.65 A/A 1.21 0.74–1.98 0.46 1.38 0.74–2.55 0.31 rs7013278 C/C T 1.00 – – 1.00 – – C/T 0.94 0.63–1.40 0.76 1.06 0.66–1.71 0.81 T/T 1.55 0.94–2.56 0.08 2.20 1.24–3.91 0.01* rs7014346 G/G A 1.00 – – 1.00 – – A/G 0.89 0.60–1.31 0.56 1.04 0.67–1.64 0.84 A/A 1.33 0.78–2.27 0.29 1.96 1.08–3.52 0.03* rs719725 C/C A 1.00 – – 1.00 – – A/C 1.29 0.69–2.39 0.43 0.93 0.47–1.86 0.84 A/A 1.07 0.57–2.01 0.84 0.80 0.39–1.62 0.53 rs10795668 G/G A 1.00 – – 1.00 – – A/G 0.79 0.54–1.16 0.23 0.82 0.51–1.31 0.40 A/A 0.54 0.25–1.17 0.11 0.82 0.37–1.83 0.62 rs3802842 A/A C 1.00 – – 1.00 – – A/C 1.53 1.05–2.23 0.03* 1.25 0.80–1.93 0.33 C/C 0.95 0.50–1.79 0.88 1.03 0.52–2.03 0.94 rs10318 C/C T 1.00 – – 1.00 – – C/T 1.10 0.74–1.64 0.63 1.20 0.73–1.90 0.50 T/T 0.50 0.12–2.03 0.33 0.62 0.15–2.55 0.50 rs4779584 C/C T 1.00 – – 1.00 – – C/T 0.92 0.63–1.34 0.65 0.72 0.45–1.14 0.16 T/T 1.02 0.52–1.98 0.96 1.01 0.46–2.22 0.98 rs4464148 T/T C 1.00 – – 1.00 – – C/T 0.78 0.54–1.15 0.21 0.74 0.48–1.16 0.19 C/C 0.95 0.50–1.80 0.87 1.39 0.67–2.90 0.38 rs4939827 C/C T 1.00 – – 1.00 – – C/T 0.90 0.60–1.34 0.59 0.82 0.52–1.30 0.40 T/T 0.82 0.50–1.36 0.44 0.81 0.45–1.48 0.50 #Potential confounding variables such as age, gender, ethnicity, stage, and treatment received were included in multivariate modeling *Statistically significant at P,0.05. associations were statistically significant [20]. In a later study by Tenesa et al, no association was found between 10 risk variants and all-cause or CRC-specific mortality after adjustment for AJCC stage, age and sex in 2838 AJCC stages I-IV CRC patients of Scottish Ancestry [21]. Likewise, we observed a similar lacked of correlation between rs10505477 (HR using multivariate analy- sis = 1.38, CI = 0.74–2.55) with survival which is in agreement with the finding by Gruber et al (HR = 1.09, CI = 0.89–1.32) [19]. For rs6983267, there was again no association with CRC survival in our study (HR using multivariate analysis = 1.35, CI = 0.73– 2.51). CRC Risk Variants Don’t Influence Clinical Outcome The HR for this SNP is in similar direction and again in agreement with previous reports by Cicek et al (HR = 1.00, CI = 0.79–1.27) and Tenesa et al (HR = 1.03, CI = 0.94–1.13) [20,21]. Furthermore, for 4 other SNPs (rs10795668, rs3802842, rs4779584 and rs4939827), the HRs obtained by Tenesa and colleagues were 0.98 (CI = 0.90–1.08), 0.93 (CI = 0.85–1.02), 0.95 (CI = 0.85–1.06) and 1.05 (CI = 0.96–1.15) respectively [21]. This result is in congruent with our study with HRs for rs10795668, rs3802842, rs4779584 and rs4939827 to be 0.82 (CI = 0.37–1.83), 1.03 (CI = 0.52–2.03), 1.01 (CI = 0.46–2.22) and 0.81 (CI = 0.45– 1.48) respectively, with the range of CI values for these SNPs overlapping between the two studies. This data thus reinforce the lack of evidence for the involvement of these variants with CRC outcome. In a recent study by Xing et al, the influence of 8 GWAS associated SNPs with CRC recurrence and survival was investi- gated in 465 Han Chinese CRC patients [22]. Two SNPs, rs4779584 (HR = 0.33, CI = 0.15–0.72 for homozygous carriers of the wild type allele (T), which is also the risk allele in GWAS, versus those who are homozygous or heterozygous carriers of the variant allele (C), the protective allele in previous GWAS) and rs10795668 (HR = 0.55, CI = 0.30–1.00 for homozygous carriers of the wild type allele (G), which is the protective allele in previous GWAS, versus those who are homozygous or heterozygous carriers of the variant allele (A), the risk allele in previous GWAS), were significantly associated with reduced risk of death and tumor recurrence respectively using a dominant genetic model [22]. For rs4779584, the result of this study (HR = 1.01, CI = 0.46–2.22) and that of Tenesa et al (HR = 0.95, CI = 0.85–1.06) [21] showed no significant influence of the risk allele of this SNP with CRC survival. This deviates from the findings of Xing et al [22]. A plausible explanation for this deviation is a much higher frequency of the risk allele among the Han Chinese compared to the population in this study and that of Tenesa and colleagues [21], which were predominately of European ancestry. CRC Risk Variants Don’t Influence Clinical Outcome associations were statistically significant [20]. In a later study by Tenesa et al, no association was found between 10 risk variants and all-cause or CRC-specific mortality after adjustment for AJCC stage, age and sex in 2838 AJCC stages I-IV CRC patients of Scottish Ancestry [21]. Likewise, we observed a similar lacked of correlation between rs10505477 (HR using multivariate analy- sis = 1.38, CI = 0.74–2.55) with survival which is in agreement with the finding by Gruber et al (HR = 1.09, CI = 0.89–1.32) [19]. For rs6983267, there was again no association with CRC survival in our study (HR using multivariate analysis = 1.35, CI = 0.73– 2.51). The HR for this SNP is in similar direction and again in agreement with previous reports by Cicek et al (HR = 1.00, CI = 0.79–1.27) and Tenesa et al (HR = 1.03, CI = 0.94–1.13) [20,21]. Furthermore, for 4 other SNPs (rs10795668, rs3802842, rs4779584 and rs4939827), the HRs obtained by Tenesa and colleagues were 0.98 (CI = 0.90–1.08), 0.93 (CI = 0.85–1.02), 0.95 (CI = 0.85–1.06) and 1.05 (CI = 0.96–1.15) respectively [21]. This result is in congruent with our study with HRs for rs10795668, rs3802842, rs4779584 and rs4939827 to be 0.82 (CI = 0.37–1.83), 1.03 (CI = 0.52–2.03), 1.01 (CI = 0.46–2.22) and 0.81 (CI = 0.45– 1.48) respectively, with the range of CI values for these SNPs overlapping between the two studies. This data thus reinforce the lack of evidence for the involvement of these variants with CRC outcome. In a recent study by Xing et al, the influence of 8 GWAS associated SNPs with CRC recurrence and survival was investi- gated in 465 Han Chinese CRC patients [22]. Two SNPs, rs4779584 (HR = 0.33, CI = 0.15–0.72 for homozygous carriers of the wild type allele (T), which is also the risk allele in GWAS, versus those who are homozygous or heterozygous carriers of the variant allele (C), the protective allele in previous GWAS) and rs10795668 (HR = 0.55, CI = 0.30–1.00 for homozygous carriers of the wild type allele (G), which is the protective allele in previous GWAS, versus those who are homozygous or heterozygous carriers of the variant allele (A), the risk allele in previous GWAS), were significantly associated with reduced risk of death and tumor recurrence respectively using a dominant genetic model [22]. CRC Risk Variants Don’t Influence Clinical Outcome recurrence while our study and that by Tenesa et al [21] evaluated overall survival and overall mortality respectively. the median follow-up period of 3.5 years may be too short especially for patients with stages I and II disease. This could have in turn resulted in a lower event rate when data from all four stages of patients were analysed together and hence led to the marginal association observed. Thirdly, information on disease free survival data which may be a better prognostic measure compared to overall survival is not available in our study. Fourthly, while we made a rigorous effort to take into consideration important clinical variables such as age, sex, ethnicity, stage of disease, site of tumor and type of treatment that may influence CRC survival in our data analysis, we however did not have information on patients’ mismatch repair status. This may have led to the combination of different types of CRC in the same group for analysis, thereby biasing the hazard estimates obtained. Lastly, the sample size of this study is moderate. This may make it underpowered to detect the association of the genetic variants with survival outcomes for two SNPs (rs719725 and rs10318) due to low allele frequencies in our population or for SNPs with small effects on survival. Thus, for rs719725 and rs10318, the results for these SNPs should be interpreted cautiously. Nonetheless, the results of this study will augment the findings of earlier studies, allowing for future meta- analysis that can further improve our understanding of the effects of these rare variants on CRC progression. This inconsistency in findings between reported studies is not surprising as the mechanisms by which these variants alter the risk of CRC is not fully understood and it is still unclear how they might influence tumor progression and patient survival. Currently, the potential functional effects of these SNPs have been most widely investigated for the variants at the 8 q24 gene locus. Since this locus is not known to encode for any gene, it was thus conceived that the variants found here may either lie within promoters or enhancer elements that can affect the transcription of genes outside this locus [10]. Supporting Information Table S1 Chromosomal loci of low-penetrance CRC susceptibility SNPs tested, their susceptibility risk allele and TaqManH SNP genotyping assay identifiers. (DOCX) Table S1 Chromosomal loci of low-penetrance CRC susceptibility SNPs tested, their susceptibility risk allele and TaqManH SNP genotyping assay identifiers. (DOCX) Table S2 Distribution of patients’ clinicopathological characteristics according to genotypes for individual SNP. (DOCX) The strength of this study is that the patients were drawn at random from 33 county areas in central and eastern NC. These regions include urban and rural areas and as such the subjects are diverse with respect to race and socioeconomic status. They are therefore more representative of a true CRC population sample compared with other studies that only include patients from a few institutions and thus have highly selected populations. Acknowledgments The authors would like to thank Mr Kevin Long for his technical assistance regarding this manuscript. CRC Risk Variants Don’t Influence Clinical Outcome However, in a study using the UCSC Browser and VISTA Enhancer database, rs7013278 and rs7014346, which showed marginal significance in our study prior to Bonferroni correction, were not found in segments containing putative enhancers or in predicted regions of regulatory potential [25]. On the other hand, rs6983267 was found to lie within a putative enhancer element that binds TCF4, a transcription factor that interacts with ß-catenin to activate the transcription of Wnt target genes, which are activated in most CRCs [26,27]. Additionally, some reports have shown that rs6983267 has a long range physical interaction with a promoter region of MYC in colorectal cancer cell lines [26,28]. Despite this, no association between the risk allele of rs6983267 and MYC expression levels has been found in normal and cancerous colon tissues [17,26,27]. Therefore, the functional consequence of rs6983267 remains uncertain. In conclusion, we observed no association between 11 CRC susceptibility variants at 6 CRC risk loci with disease outcome in our study population, suggesting little influence of these SNPs on CRC progression. For rs10795668 at 10 p14, rs3802842 at 11 q23 and rs 4779584 at 15 q13, a systematic search by Niittymaki and colleagues failed to show any association of these SNPs with predicted enhancer or regulatory elements [29]. Further investigation using tumor samples again showed a lack of allelic imbalance between the risk allele of these SNPs with CRC, prompting the authors to conclude that these risk variants were unlikely somatically selected for neoplastic progression [29]. While the functional effects of these susceptibility SNPs remain to be further validated, the results of these functional studies to date support our finding that the majority of low-penetrance CRC variants are involved in initiation rather than progression of CRC. 6. Lichtenstein P, Holm NV, Verkasalo PK, Iliadou A, Kaprio J, et al. (2000) Environmental and heritable factors in the causation of cancer: analyses of Author Contributions Conceived and designed the experiments: JMH PO TOK CFM RSS HLM. Performed the experiments: JMH PO CAC MW. Analyzed the data: JMH PO. Wrote the paper: JMH PO. Performed statistical analyses: JAG. Conceived and designed the experiments: JMH PO TOK CFM RSS HLM. Performed the experiments: JMH PO CAC MW. Analyzed the data: JMH PO. Wrote the paper: JMH PO. Performed statistical analyses: JAG. There are however some limitations in our study. Firstly, the limited sample sizes of certain stages prevented a more detailed subgroup analysis. At such, it is possible that associations restricted to patients from certain stages may have been missed. Secondly, 10. Haiman CA, Le Marchand L, Yamamato J, Stram DO, Sheng X, et al. (2007) A common genetic risk factor for colorectal and prostate cancer. Nat Genet 39: 954–956. 9. Gruber SB, Moreno V, Rozek LS, Rennerts HS, Lejbkowicz F, et al. (2007) Genetic variation in 8 q24 associated with risk of colorectal cancer. Cancer Biol Ther 6: 1143–1147. 5. Rodrigues NR, Rowan A, Smith ME, Kerr IB, Bodmer WF, et al. (1990) p53 mutations in colorectal cancer. Proc Natl Acad Sci USA 87: 7555–7559. 7. Collins FS, Guyer MS, Charkravarti A (1997) Variations on a theme: cataloging human DNA sequence variation. Science 278: 1580–1581. 1. O’Connell JB, Maggard MA, Ko CY (2004) Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst 96: 1420–1425. CRC Risk Variants Don’t Influence Clinical Outcome While the allele frequencies of rs4779584 in the Han Chinese CRC patients were not directly reported in the paper by Xing et al [22], a calculation of the allele frequencies of this population based on the genotype of patients presented showed that the calculated allele frequencies (C = 0.19 and T = 0.81) were congruent with that from the Chinese populations in the Hapmap (CHB: C = 0.18 and T = 0.82; CHD: C = 0.16 and T = 0.84) and the Han Chinese population that we have previously genotyped (C = 0.17 and T = 0.83) as part of the four additional populations used for genotyping control. Based on the data from Hapmap and our data of the four additional populations genotyped (data not shown), the allele frequency for the risk allele (T) of rs4779584 was highest in the Han Chinese, followed by African-American and lastly European-American. For rs10795668, HR for overall survival in our study and that by Tenesa et al [21] were 0.82 (CI = 0.37–1.83) and 0.98 (CI = 0.90–1.08) respectively. This divergence in study results to that of Xing et al is likely attributed to methodological differences between studies rather than due to differences in allele frequencies as the study by Xing et al [22] evaluated CRC PLoS ONE \| www.plosone.org July 2012 \| Volume 7 \| Issue 7 \| e41954 4 2. Fearon ER, Vogelstein B (1990) A genetic model for colorectal tumorigenesis. Cell 61: 759–767. 8. Broderick P, Carvajal-Carmona L, Pittman AM, Webb E, Howarth K, et al. (2007) A genome-wide association study shows that common alleles of SMAD7 influence colorectal cancer risk. Nat Genet 39: 1315–1317. cohorts of twins from Sweden, Denmark, and Finland. N Engl J Med 343: 78– 85. cohorts of twins from Sweden, Denmark, and Finland. N Engl J Med 343: 78– 85. 4. Kinzler KW, Vogelstein B (1996) Lessons from hereditary colorectal cancer. Cell 87: 159–170. 3. Greenblatt MS, Bennett WP, Hollstein M, Harris CC (1994) Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 54: 4855–4878. CRC Risk Variants Don’t Influence Clinical Outcome CRC Risk Variants Don’t Influence Clinical Outcome 11. Jaeger E, Webb E, Howarth K, Carvajal-Carmona L, Rowan A, et al. (2008) Common genetic variants at the CRAC1 (HMPS) locus on chromosome 15 q13.3 influence colorectal cancer risk. Nat Genet 40: 26–28. 20. Cicek MS, Slager SL, Achenbach SJ, French AJ, Blair HE, et al. (2009) Functional and clinical significance of variants localized to 8 q24 in colon cancer. Cancer Epidemiol Biomarkers Prev 18: 2492–2500. 21. Tenesa A, Theodoratou E, Din FVN, Farrington SM, Cetnarskyj R, et al. (2010) Ten common genetic variants associated with colorectal cancer risk are not associated with survival after diagnosis. Clin Cancer Res 16: 3754–3759. 12. Li L, Plummer SJ, Thompson CL, Merkulova A, Acheson LS, et al. (2008) A common 8 q24 variant and the risk of colon cancer: a population-based case- control study. Cancer Epidemiol Biomarkers Prev 17: 339–342. control study. Cancer Epidemiol Biomarkers Prev 17: 339–342. 22. Xing J, Myers RE, He X, Qu F, Zhou F, et al. (2011) GWAS-identified colorectal cancer susceptibility locus associates with disease prognosis. Eur J Cancer 47: 1699–1707. 13. Poynter JN, Figueiredo JC, Conti DV, Kennedy K, Gallinger S, et al. (2007) Variants on 9 p24 and 8 q24 are associated with risk of colorectal cancer: results from the Colon Cancer Family Registry. Cancer Res 67: 11128–11132. 23. Ayanian JZ, Chrischilles EA, Fletcher RH, Fouad MN, Harrington DP, et al. (2004) Understanding cancer treatment and outcomes: the Cancer Care Outcomes Research and Surveillance Consortium. J Clin Oncol 22: 2992–2996. 14. Tenesa A, Farrington SM, Prendergast JG, Porteous ME, Walker M, et al. (2008) Genome-wide association scan identifies a colorectal cancer susceptibility locus on 11 q23 and replicates risk loci at 8 q24 and 18 q21. Nature Genet 40: 631–637. Marsh S, King CR, Ahluwalia R, McLeod HL (2004) Distribution 24. Marsh S, King CR, Ahluwalia R, McLeod HL (2004) Distribution of IT P32T alleles in multiple world populations. J Hum Genet 49: 579–581. P32T alleles in multiple world populations. J Hum Genet 49: 579–58 15. Tomlinson IP, Webb E, Carvajal-Carmona L, Broderick P, Howarth K, et al. (2008) A genome-wide association study identifies colorectal cancer susceptibility loci on chromosomes 10 p14 and 8 q23.3. Nature Genet 40: 623–630. 25. Yeager M, Xiao N, Hayes RB, Bouffard P, Desany B, et al. References cohorts of twins from Sweden, Denmark, and Finland. N Engl J Med 343: 78– 85. 7. Collins FS, Guyer MS, Charkravarti A (1997) Variations on a theme: cataloging human DNA sequence variation. Science 278: 1580–1581. 8. Broderick P, Carvajal-Carmona L, Pittman AM, Webb E, Howarth K, et al. (2007) A genome-wide association study shows that common alleles of SMAD7 influence colorectal cancer risk. Nat Genet 39: 1315–1317. 3. Greenblatt MS, Bennett WP, Hollstein M, Harris CC (1994) Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 54: 4855–4878. 9. Gruber SB, Moreno V, Rozek LS, Rennerts HS, Lejbkowicz F, et al. (2007) Genetic variation in 8 q24 associated with risk of colorectal cancer. Cancer Biol Ther 6: 1143–1147. 4. Kinzler KW, Vogelstein B (1996) Lessons from hereditary colorectal cancer. Cell 87: 159–170. 10. Haiman CA, Le Marchand L, Yamamato J, Stram DO, Sheng X, et al. (2007) A common genetic risk factor for colorectal and prostate cancer. Nat Genet 39: 954–956. PLoS ONE \| www.plosone.org July 2012 \| Volume 7 \| Issue 7 \| e41954 5 PLoS ONE \| www.plosone.org CRC Risk Variants Don’t Influence Clinical Outcome (2008) Comprehensive resequence analysis of 136 kb region of human chromosome 8 q24 associated with prostate and colon cancer. Hum Genet 124: 161–170. 16. Tuupanen S, Niittymaki I, Nousiainen K, Vanharanta S, Mecklin JP, et al. (2008) Allelic imbalance at rs6983267 suggests selection of the risk allele in somatic colorectal tumor evolution. Cancer Res 68: 14–17. 26. Pomerantz MM, Ahmadiyeh N, Jia L, Herman P, Verzi MP, et al. (2009) The 8 q24 cancer risk variant rs6983267 shows long-range interaction with MYC in colorectal cancer. Nat Genet 41: 882–884. 17. Zanke BW, Greenwood CM, Rangrej J, Kustra R, Tenesa A, et al. (2007) Genome-wide association scan identifies a colorectal cancer susceptibility locus on chromosome 8 q24. Nature Genet 39: 989–994. 27. Tuupanen S, Turunen M, Lehtonen R, Hallikas O, Vanharanta S, et al. (2009) The common colorectal cancer predisposition SNP rs6983267 at chromosome 8 q24 confers potential to enhanced Wnt signaling. Nat Genet 41: 161–170. 18. Houlston RS, Webb E, Broderick P, Pittman AM, Di Bernado MC, et al. (2008) Meta-analysis of genome-wide association data identifies four new susceptibility loci for colorectal cancer. Nat Genet 40: 1426–1435. q p g g 28. Wright JB, Brown SJ, Cole MD (2010) Upregulation of c-MYC in cis through a large chromatin loop linked to a cancer risk-associated SNP in colorectal cancer cells. Mol Cell Biol 30: 1411–1420. 19. Gruber SB, Moreno V, Rozek LS, Rennert HS, Lejbkowicz F, et al. (2007) Genetic variation in 8 q24 associated with risk of colorectal cancer. Cancer Biol Ther 6: 1143–1147. 29. Niittymaki I, Tuupanen S, Li Y, Jarvinen H, Mecklin J, et al. (2011) Systematic search for enhancer elements and somatic allelic imbalance at seven low- penetrance colorectal cancer predisposition loci. BMC Med Genet 12: 23. PLoS ONE \| www.plosone.org July 2012 \| Volume 7 \| Issue 7 \| e41954 6	8,172
https://openalex.org/W3043459603_1	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	8,827	15,481	TESIS DOCTORAL Análisis y mejora de la fluidodinámica en estaciones depuradoras de aguas residuales Francisco Sánchez Fernández Ingeniero Industrial dirigida por: Dr. Antonio Viedma Robles Dr. Antonio Sánchez Kaiser Doctorado en Tecnologías Industriales Departamento de Ingeniería Térmica y de Fluidos Universidad Politécnica de Cartagena Septiembre de 2019 Este trabajo ha sido financiado por la Fundación Séneca – Agencia de Ciencia y Tecnología de la Región de Murcia, perteneciente a la Consejería de a la Consejería de Industria, Turismo, Empresa e Innovación de la Comunidad Autónoma de la Región de Murcia (CARM), por medio del programa Contratos Predoctorales de Formación de Personal investigador (19778/FPI/15). Resumen La investigación realizada en esta Tesis Doctoral se centra en el análisis y mejora de los procesos que tienen lugar en el interior de instalaciones de depuración de aguas residuales. Tradicionalmente, el diseño de las instalaciones de las estaciones de depuración de aguas residuales (EDAR) se hace tomando consideraciones y simplificaciones del flujo, debido principalmente a la complejidad de este y a la falta de herramientas para su análisis. Sin embargo, los fenómenos fluidodinámicos locales juegan un papel importante en la depuración, pudiendo existir ineficiencias tales como zonas de paso rápido, muertas o sin renovar. Es necesario comprender el comportamiento del flujo en el interior de las instalaciones para un correcto diseño de estas, que asegure un correcto tratamiento sin un excesivo coste energético. En este trabajo, se han analizado distintas instalaciones de depuración, tanto del tratamiento secundario (reactor biológico) como terciario (canal ultravioleta UV) de la EDAR. Se han empleado herramientas de mecánica de fluidos computacional (CFD) como base para el análisis, aunque también se han desarrollado y empleado procedimientos experimentales y analíticos para la validación de los modelos numéricos. Los análisis se han realizado sobre instalaciones reales concretas de EDAR, generalizando posteriormente los resultados para que sean de aplicación universal. El alcance de los análisis realizados comprende la fluidodinámica de la instalación y otros fenómenos de distinta naturaleza como es el caso de los procesos de transferencia de oxígeno y reacciones biológicas en el interior de un reactor biológicos, o los fenómenos de propagación de luz UV e inactivación de microorganismos en un canal UV. Los resultados alcanzados proporcionan aspectos característicos de los fenómenos fluidodinámicos que ocurren en reactores biológicos y canales UV de EDAR, así como su interacción con los fenómenos biológicos y radiativos que se producen en la instalación, bajo distintas condiciones de operación. Como parte de la discusión de los resultados se presentan las ineficiencias fluidodinámicas detectadas más significativas, así como algunas propuestas de modificación geométrica destinadas a la corrección de estas. De manera complementaria, durante el desarrollo de la investigación, se han empleado y desarrollado diferentes metodologías y aproximaciones, principalmente numéricas, aunque también analíticas y experimentales. Se ha llevado a cabo un análisis sobre la aplicabilidad, potencial y limitaciones de dichas metodologías y aproximaciones en función del tipo y características del flujo analizado, lo que también supone una contribución al estudio fluidodinámico de instalaciones de EDAR. Abstract The research carried out in this Doctoral Thesis focuses in the analysis and improvement of the processes that occurs within wastewater treatment plant (WWTP) facilities. Traditionally, the design of these facilities considers some simplifications of the flow. However, fluid dynamic processes play an important role in water treatment; it is possible to have inefficiencies such as dead zones or short-circuiting flows. It is necessary to understand the flow behaviour within the facilities for a correct design, which ensures a proper treatment without an excessive energy consumption. The research involves the analysis of different WWTP facilities: biological reactors and ultraviolet (UV) channels. The analyses have been done mainly by means of Computational Fluid Dynamics (CFD) tools. Analytical models and experimental procedures have been also employed for validation of the numerical models. The numerical models are applied to specific actual plants, generalizing the results so that they are universally applicable. Apart from the hydraulic processes, the scope of the work involves other phenomena that take place in WWTP facilities: oxygen transference and biological reactions within biological reactors, UV light propagation and microbial inactivation in UV channels. The results obtained provide relevant information about the hydraulic processes that occurs within biological reactors and UV channels, as well as their interaction with biological and radiative phenomena, under different operational conditions. Some of the most relevant inefficiencies detected, as well as geometrical modifications intended to correct them, are included in the text. Additionally, different methodologies and procedures (mainly numerical) have been employed and developed during the research. A discussion about the applicability, potential and limitations of these methodologies on the basis of type and features of the flow is carried out. This discussion constitutes also a contribution to the field of hydraulic processes in WWTPs. Agradecimientos No es fácil reconocer en unas pocas líneas a todas las personas que han contribuido a la realización de esta Tesis Doctoral, es por ello que pido disculpas si se me olvida mencionar a alguna de las tantas personas que han puesto su granito de arena en este proyecto. A mi director Antonio Viedma Robles, por todo lo que me ha enseñado y transmitido durante estos años, la confianza que ha depositado en mí y todo lo que me ha ayudado, incluso en mis proyectos personales. A mi director Antonio Sánchez Kaiser, por toda la confianza que ha depositado en mí, por todo lo que me ha enseñado, especialmente su visión a la hora de resolver problemas y buscar oportunidades, y por todo lo que se ha implicado en mi desarrollo como investigador, como ingeniero y, sobre todo, como persona. A mis compañeros de departamento Abraham, Carmen, Javi, Pablo, Guillermo, Iván y David por todos los consejos, momentos y conversaciones que hemos compartido durante esta etapa. A mi familia, mis padres y mi hermana, por darme todo su apoyo y empuje durante estos años. A Tamara, sin ella todo hubiera sido mucho más difícil. A la Entidad de Saneamiento y Depuración de Aguas Residuales de la Región de Murcia (ESAMUR), por financiar parte de la investigación y prestar sus instalaciones para realizar las diferentes medidas y experimentos. En cierta manera, también les agradezco que me introdujeran en este mundo. A Fernando, Reyes y el resto de trabajadores de las depuradoras de San Pedro del Pinatar y Los Alcázares, por colaborar y prestar su ayuda en las instalaciones. A la Fundación Séneca, por financiar la investigación, y por el trato y atención recibida, especialmente por parte de Viviane Barelli. A mi gran amigo y socio Fran Nicolás Pérez, por su forma de ser, todo lo que me ha enseñado, y compartir conmigo ese gran proyecto llamado Lynx Simulations, que da continuidad a todo lo hecho durante estos años, y del que estoy muy orgulloso. I Índice general 1. Introducción ......................................................................................... 1 1.1. Origen del estudio ................................................................................................ 1 1.2. Estado del arte ...................................................................................................... 5 1.2.1. CFD en el análisis de reactores biológicos de EDAR .................................................... 7 1.2.2. CFD en el análisis de sistemas de desinfección UV ................................................... 13 1.3. Identificación de objetivos.................................................................................. 15 1.4. Estructura de la memoria de tesis ...................................................................... 16 2. Aspectos del estudio y descripción de las instalaciones ..................... 19 2.1. Descripción del reactor biológico analizado fluidodinámicamente ................... 19 2.2. Descripción del reactor biológico analizado a nivel fluidodinámico y biológico.22 2.3. Descripción del canal UV estudiado ................................................................... 25 3. Modelización numérica de los procesos de depuración ..................... 29 3.1. Ecuaciones de comportamiento ......................................................................... 29 3.1.1. Fluidodinámica. Ecuaciones de Navier Stokes. Turbulencia. Flujo multifásico. ........ 29 3.1.2. Transporte de componentes. Cálculo RTD ................................................................ 35 3.1.3. Reacciones biológicas. Modelo ASM1....................................................................... 35 3.1.4. Radiación UV. Ecuación del transporte radiativo. Distribución de dosis UV. Desinfección de microorganismos. ........................................................................... 39 3.2. Dominio computacional y mallado ..................................................................... 41 3.2.1. Reactor biológico de la EDAR de San Pedro del Pinatar ........................................... 41 3.2.2. Reactor biológico de la EDAR a remodelar ............................................................... 43 3.2.3. Canal UV de la EDAR de Los Alcázares ...................................................................... 45 3.3. Procedimiento de simulación numérica ............................................................. 47 3.3.1. Análisis fluidodinámico del reactor biológico de una EDAR ..................................... 47 3.3.2. Análisis fluidodinámico y biológico del reactor biológico de una EDAR ................... 50 3.3.3. Análisis de la desinfección en el canal UV de una EDAR ........................................... 53 4. Análisis fluidodinámico del reactor biológico de una EDAR ................ 58 4.1. Análisis del flujo en el interior del reactor. Campos de velocidad ..................... 59 II Índice general 4.2. Evaluación de las herramientas de análisis RTD en reactores biológicos .......... 68 4.2.1. Fundamentos del análisis RTD .................................................................................. 68 4.2.2. Diseño del experimento de trazador de tipo pulso .................................................. 71 4.2.3. Desarrollo del experimento. Resultados experimentales ......................................... 75 4.2.4. Análisis de los resultados experimentales. Modelización de la curva RTD en sistemas con recirculación ....................................................................................................... 77 4.2.5. Selección de la técnica CFD más adecuada para la simulación RTD ......................... 87 4.3. Análisis RTD del reactor biológico ...................................................................... 93 4.4. Mejora de la fluidodinámica en el interior del reactor ...................................... 94 4.4.1. Modificaciones propuestas ....................................................................................... 96 4.4.2. Resultados de flujo con las modificaciones propuestas ........................................... 99 4.4.3. Análisis RTD de las modificaciones propuestas....................................................... 109 4.4.4. Conclusiones ........................................................................................................... 111 5. Análisis fluidodinámico y biológico del reactor biológico de una EDAR 115 5.1. Análisis del flujo en el interior del reactor. Campos de velocidad ................... 115 5.1.1. Campos de velocidad en el reactor ......................................................................... 116 5.1.2. Fracción volumétrica de aire y nivel de mezclado .................................................. 120 5.1.3. Análisis RTD del reactor .......................................................................................... 122 5.2. Análisis de los procesos biológicos en el interior del reactor .......................... 124 5.2.1. Simulación biológica mediante herramientas de simulación de procesos ............. 124 5.2.2. Simulación biológica mediante herramientas CFD ................................................. 127 5.2.3. Evaluación del consumo energético de la aireación ............................................... 133 5.3. Evaluación energética del sistema de aireación bajo distintas configuraciones y regímenes de aireación..................................................................................... 135 5.3.1. Influencia del sistema de aireación en el consumo energético .............................. 135 5.3.2. Influencia del régimen de aireación en el consumo energético ............................. 140 5.4. Conclusiones ..................................................................................................... 144 6. Análisis de la de la desinfección en el canal UV de una EDAR ........... 147 6.1. Análisis del funcionamiento del canal: fluidodinámica, radiación, distribución de dosis UV e inactivación de microorganismos ................................................... 148 6.1.1. Comportamiento del flujo en el canal. Campo de velocidad .................................. 148 6.1.2. Campo de intensidad de radiación UV .................................................................... 151 6.1.3. Distribución de dosis UV ......................................................................................... 152 6.1.4. Inactivación de microorganismos en el canal ......................................................... 154 III 6.2. Validación experimental del modelo numérico ............................................... 157 6.2.1. Validación experimental del campo de velocidades en el canal............................. 157 6.2.2. Validación del campo de intensidad UV.................................................................. 164 6.3. Estudio de la influencia de la placa perforada y del banco .............................. 168 6.3.1. Análisis del funcionamiento con un solo banco de lámparas activado .................. 168 6.3.2. Influencia de la placa perforada en la inactivación ................................................. 171 6.4. Análisis de la influencia de las condiciones de operación: caudal, transmitancia y potencia UV. Optimización del proceso ........................................................... 174 6.4.1. Influencia del caudal ............................................................................................... 174 6.4.2. Influencia de la transmitancia ................................................................................. 176 6.4.3. Potencia UV mínima para conseguir un nivel de inactivación requerido ............... 178 6.5. Análisis bajo condiciones de fallo ..................................................................... 180 6.5.1. Fallo en alguna de las lámparas del canal ............................................................... 180 6.5.2. Fallo en el sistema de control de control de la superficie libre .............................. 184 6.6. Análisis de mejoras destinadas a mejorar la eficiencia del canal ..................... 186 6.6.1. Placa helicoidal a la entrada del canal .................................................................... 186 6.6.2. Reducción del margen superficie libre – lámpara................................................... 189 7. Conclusiones .................................................................................... 193 7.1. Conclusiones del análisis fluidodinámico del reactor biológico de una EDAR. 194 7.2. Conclusiones del análisis fluidodinámico y biológico del reactor biológico de una EDAR ................................................................................................................. 195 7.3. Conclusiones del análisis de la desinfección en el canal UV de una EDAR....... 197 7.4. Revisión de los objetivos, aportaciones científicas y futuras líneas de trabajo 199 Referencias bibliográficas....................................................................... 201 V Índice de figuras Figura 1. Vista general de la EDAR de Aspe (Alicante). ................................................................. 3 Figura 2. Línea de proceso de una EDAR convencional. ............................................................... 4 Figura 3. Simulación CFD de un canal de oxidación biológico. ..................................................... 9 Figura 4. Figuras de la publicación de Le Moullec et al. (2008). ................................................. 10 Figura 5. Vista general de la EDAR de San Pedro del Pinatar. ..................................................... 20 Figura 6. Detalle de los difusores de aire sobre el suelo de la zona óxica 3 del reactor............. 20 Figura 7. Plano del reactor biológico de la EDAR de San Pedro del Pinatar ............................... 21 Figura 8. Plano del reactor biológico sobre el que se realiza el estudio. .................................... 22 Figura 9. Esquema conceptual del tratamiento secundario de la EDAR. .................................... 23 Figura 10. Distribución de los difusores de aire bajo las dos configuraciones a analizar ........... 25 Figura 11. Fotografía del canal UV. ............................................................................................. 26 Figura 12. a) Fotografía de un módulo UV; b) distribución de las lámparas ............................... 26 Figura 13. Plano del canal UV (y el canal previo al mismo) de la EDAR Los Alcázares ................ 27 Figura 14. Detalles de la placa perforada (a) y vertedero de separación entre los canales (b) .. 27 Figura 15. Diagrama conceptual de los componentes y reacciones en el modelo ASM1. ......... 37 Figura 16. Curvas de inactivación frente a dosis UV en agua residual de distintos organismos. 41 Figura 17. Geometría del modelo CFD del reactor biológico de San Pedro del Pinatar. ............ 41 Figura 18. Comparación del modelo del agitador (a) y la geometría real del mismo (b). .......... 42 Figura 19. Mallado del reactor biológico de San Pedro del Pinatar. ........................................... 42 Figura 20. Geometría del reactor biológico de la EDAR a remodelar. ........................................ 43 Figura 21. Mallado de la geometría del reactor biológico a remodelar. .................................... 44 Figura 22. Detalle del mallado en la zona del agitador (a) y los difusores de aire (b). ............... 44 Figura 23. Detalle del refinamiento de la malla sobre el tabique que separa las zonas............. 45 Figura 24. Detalle de la superficie de control de entrada al dominio del canal UV. ................... 45 Figura 25. a) Vista general de la geometría del canal UV; b) vista en detalle del canal UV. ....... 46 Figura 26. Mallado en el entorno de las lámparas UV y los soportes de modulo.. .................... 47 Figura 27. Procedimiento de simulación para cada uno de los escenarios analizados en el estudio “Análisis fluidodinámico del reactor biológico de una EDAR”. ................................................... 48 Figura 28. Procedimiento de simulación para cada uno de los escenarios analizados en el estudio “Análisis fluidodinámico y biológico del reactor biológico de una EDAR”. ................................. 51 Figura 29. Procedimiento de simulación para cada uno de los escenarios analizados en el estudio “Análisis de la desinfección en el canal UV de una EDAR”.......................................................... 54 Figura 30. Simulación 2D para el cálculo de la altura de la superficie de entrada. .................... 55 Figura 31. Contorno de velocidad en el caso invierno sin aireación .......................................... 60 Figura 32. Vectores velocidad en el caso verano sin aireación. .................................................. 60 Figura 33. Campo de velocidad en los casos sin aireación (a) y con aireación (b). .................... 61 Figura 34. Contorno de velocidad en el caso invierno sin aireación (a) y con aireación (b). ...... 62 Figura 35. Presión reducida en el caso invierno con aireación ................................................... 63 Figura 36. Histograma de velocidad para el caso verano sin aireación. ..................................... 63 Figura 37. Histograma de velocidad para el caso verano con aireación. .................................... 64 Figura 38. Histograma de velocidad para el caso invierno sin aireación. ................................... 64 VI Índice de figuras Figura 39. Histograma de velocidad para el caso invierno con aireación. .................................. 64 Figura 40. Zonas con velocidad menor de 10 cm/s en el caso verano sin aireación .................. 65 Figura 41. Zonas con velocidad menor de 10 cm/s en el caso verano con aireación. ................ 66 Figura 42. Zonas con velocidad menor de 10 cm/s en el caso invierno sin aireación ................ 66 Figura 43. Zonas con velocidad menor de 10 cm/s en el caso invierno con aireación ............... 66 Figura 44. Zonas con velocidad mayor de 50 cm/s en el caso verano sin aireación................... 67 Figura 45. Zonas con velocidad mayor de 50 cm/s en el caso verano con aireación. ................ 67 Figura 46. Ejemplo de curvas RTD en un depósito de agua ........................................................ 69 Figura 47. Formas típicas de las curvas E para distintos valores de tiempo y dispersión. .......... 70 Figura 48. Descripción gráfica del significado de la dispersión d en la curva RTD. ..................... 71 Figura 49. Curva E estimada mediante una simulación CFD del reactor. ................................... 73 Figura 50. Concentración de Li+ estimada para Q=1200 m3/h. Introduciendo 5.33 kg de Li+. .. 74 Figura 51. Esquema de la planta y puntos donde se tomaron las muestras experimentales. ... 75 Figura 52. Evolución del caudal tratado, recirculado y total durante el ensayo......................... 76 Figura 53. Concentración de Li+ de las muestras tomadas en la salida y la recirculación.......... 77 Figura 54. Curva RTD obtenida en la salida del reactor biológico durante el ensayo................. 78 Figura 55. Suma de los cuadrados de los residuales entre salida y recirculación. ...................... 80 Figura 56. Concentraciones en la salida y recirculación tras aplicar un adelanto de 42.6 min .. 80 Figura 57. Fracción de Li+ trasegada en salida, recirculación y pretratamiento. ....................... 81 Figura 58. Comparación de las curvas teórica y experimental, usando los datos de concentración en la recirculación. ...................................................................................................................... 83 Figura 59. Comparación de las curvas teórica y experimental, suponiendo igual concentración en salida y recirculación. ............................................................................................................. 84 Figura 60. Esquema del modelo analítico con tres entradas independientes. ........................... 84 Figura 61. Comparación de las curvas teórica y experimental, con el método de las tres entradas independientes. .......................................................................................................................... 85 Figura 62. Comparación de las curvas RTD obtenidas por los tres métodos desarrollados y aplicando directamente el modelo de Levenspiel con la curva experimental............................ 86 Figura 63. Vectores velocidad en el caso de validación experimental........................................ 87 Figura 64. Comparación de la curva RTD obtenida por las dos técnicas CFD y el experimento. 88 Figura 65. Influencia del parámetro CL en la curva calculada con seguimiento de partículas.. .. 90 Figura 66. Campo F en el reactor para un caso sin aireación.. ................................................... 90 Figura 67. Influencia del tamaño de elemento en la curva RTD calculada con seguimiento de partículas.. ................................................................................................................................... 91 Figura 68. Comparación entre las curvas RTD obtenidas experimentalmente y numéricamente mediante la resolución de la ecuación de transporte de un trazador inerte. ............................ 92 Figura 69. Curvas E obtenidas para las diferentes condiciones de caudal y aireación ............... 94 Figura 70. Vista lateral de la configuración 4. ............................................................................. 96 Figura 71. Disposición de los 4 nuevos orificios practicados para la configuración 2. ............... 98 Figura 72. Vista lateral de la configuración 3. ............................................................................. 98 Figura 73. Geometría de la configuración 4.. .............................................................................. 99 Figura 74. Velocidad para el caso sin aireación con la configuración 2. ................................... 100 Figura 75. Vectores velocidad para el caso con aireación con la configuración 2. ................... 100 Figura 76. Zonas con velocidad mayor de 50 cm/s para el caso sin aireación con la conf 2. ... 101 Figura 77. Zonas con velocidad mayor de 50 cm/s para el caso con aireación con la conf 2. .. 101 Figura 78. Histograma de velocidad para el caso sin aireación con configuración 2. ............... 102 Figura 79. Histograma de velocidad para el caso con aireación con configuración 2. ............. 102 Figura 80. Zonas con velocidad menor de 10 cm/s para el caso sin aireación con conf 2........ 103 VII Figura 81. Zonas con velocidad menor de 10 cm/s para el caso con aireación con conf 2. ..... 103 Figura 82. Velocidad en los casos sin (a) y con aireación (b) con la conf. 3. ............................. 104 Figura 83. Histograma de velocidad para el caso sin aireación con configuración 3. ............... 105 Figura 84. Histograma de velocidad para el caso con aireación con configuración 3. ............. 105 Figura 85. Zonas con velocidad menor de 10 cm/s para el caso sin aireación con conf 3........ 106 Figura 86. Zonas con velocidad menor de 10 cm/s para el caso con aireación con conf 3. ..... 106 Figura 87. Vectores velocidad en el plano z=2.5 m para el caso sin aireación con la conf 4. ... 107 Figura 88. Zonas con velocidad menor de 10 cm/s para el caso sin aireación con conf 4........ 107 Figura 89. Zonas con velocidad menor de 10 cm/s para el caso con aireación con conf 4. ..... 108 Figura 90. Histogramas de velocidad obtenidos para condiciones de no aireación. ................ 108 Figura 91. Histogramas de velocidad obtenidos para condiciones de aireación. ..................... 109 Figura 92. Comparación de las curvas RTD para el caso sin aireación. ..................................... 110 Figura 93. Comparación de las curvas RTD para el caso con aireación. ................................... 111 Figura 94. Campo de velocidad en el reactor, bajo condiciones de aireación (conf. C1). ........ 116 Figura 95. Líneas de corriente en la zona anóxica del reactor (conf C1). ................................. 117 Figura 96. Líneas de corriente en la zona óxica bajo condiciones de aireación (conf. C1). ...... 117 Figura 97. Detalle del campo de velocidades en torno a las columnas de aireación (conf C1).118 Figura 98. Presión reducida en el reactor bajo condiciones de aireación (conf. C1).. .............. 118 Figura 99. Líneas de corriente en la zona óxica, condiciones de no aireación (conf. C1). ........ 119 Figura 100. Volumen del reactor con velocidad menor de 10 cm/s bajo condiciones de no aireación (conf. C1). .................................................................................................................. 119 Figura 101. Volumen del reactor con velocidad menor de 10 cm/s bajo condiciones de aireación (conf. C1).. ................................................................................................................................. 120 Figura 102. Contornos de difusividad turbulenta en el reactor biológico (conf. C1) bajo condiciones de no aireación (a) y aireación (b).. ...................................................................... 121 Figura 103. Fracción volumétrica de aire en la zona óxica del reactor (conf. C1). ................... 122 Figura 104. Curvas RTD para el ensayo de tipo pulso (E) en el reactor biológico bajo condiciones de aireación y no aireación. ...................................................................................................... 123 Figura 105. Esquema y condiciones de funcionamiento del modelo del reactor biológico a simular mediante herramientas de simulación de procesos. ................................................... 125 Figura 106. Evolución de la concentración de oxígeno disuelto en el efluente de la zona óxica (conf. C1) con la simulación mediante herramientas de simulación de procesos. ................... 126 Figura 107. Evolución de la concentración de amonio en el efluente de la zona óxica (conf. C1) con la simulación mediante herramientas de simulación de procesos. ................................... 127 Figura 108. Evolución de la concentración de nitratos en el efluente de la zona óxica (conf. C1) con la simulación mediante herramientas de simulación de procesos. ................................... 127 Figura 109. Campo de coeficiente de transferencia KLa en el reactor biológico (conf. C1). ..... 128 Figura 110. Concentración de sustrato biodegradable soluble (SS) en el reactor (conf. C1). ... 129 Figura 111.Concentración de biomasa heterótrofa en el reactor (XBH). ................................... 129 Figura 112. Evolución de las concentraciones de oxígeno disuelto, nitratos y amonio en el efluente del reactor obtenidas mediante la simulación CFD-ASM1. ........................................ 130 Figura 113. Campo de SO en la zona óxica durante el ciclo de aireación (conf. C1)................. 131 Figura 114. Campo de SNH en la zona óxica durante el ciclo de aireación (conf. C1).. .............. 131 Figura 115. Distribución del ratio de la reacción R1 en la zona óxica durante el ciclo de aireación (conf. C1). .................................................................................................................................. 132 Figura 116. Evolución del oxígeno en el efluente, masa total de SO en el reactor y eficiencia en la transferencia de oxígeno (SOTE) durante un ciclo de aireación (conf. C1).. ......................... 134 Figura 117. Campo de velocidad en el reactor, funcionando con la configuración C2. ............ 136 VIII Índice de figuras Figura 118. Fracción volumétrica de aire en la zona óxica bajo la configuración C2. ............... 137 Figura 119. Evolución de las concentraciones de oxígeno disuelto, nitratos y amonio en el efluente del reactor con la configuración de aireación C2. ...................................................... 138 Figura 120. Campo de SO en la zona óxica durante el ciclo de aireación con conf C2.. ............ 138 Figura 121. Comparación de la evolución del oxígeno en el efluente, masa total de SO en el reactor y eficiencia en la transferencia de oxígeno (SOTE) durante un ciclo de aireación para las dos configuraciones de aireación analizadas. ........................................................................... 139 Figura 122. Campo de velocidad en el reactor, funcionando con la configuración C2b. .......... 141 Figura 123. Fracción volumétrica de aire en la zona óxica del reactor, configuración C2b. ..... 141 Figura 124. Evolución de las concentraciones de oxígeno disuelto, nitratos y amonio en el efluente del reactor con la configuración de aireación C2b. .................................................... 142 Figura 125. Campo de SO en la zona óxica durante el ciclo de aireación, conf C2b.. ................ 143 Figura 126. Comparación de la evolución del oxígeno en el efluente, masa total de SO en el reactor y eficiencia en la transferencia de oxígeno (SOTE) durante un ciclo de aireación para las tres configuraciones de aireación analizadas ........................................................................... 144 Figura 127. Líneas de corriente en la balsa previa al canal UV. ................................................ 148 Figura 128. Vectores velocidad en un plano vertical perpendicular al canal, 100 mm antes de la placa perforada.. ....................................................................................................................... 149 Figura 129. Vectores velocidad en un plano vertical perpendicular al canal, 100 mm después de la placa perforada. .................................................................................................................... 149 Figura 130. Líneas de corriente en el interior del canal UV. ..................................................... 150 Figura 131. Campos de velocidad en los planos horizontales del canal localizados a 65 mm de altura (a) y 455 mm sobre el fondo (b). .................................................................................... 151 Figura 132. Campo de intensidad UV en el entorno de las lámparas.. .................................... 152 Figura 133. Vista cenital del campo de intensidad UV en el canal............................................ 152 Figura 134. Posición y dosis UV adquirida por la nube de partículas tras 31 s. ....................... 153 Figura 135. Histograma de dosis UV para las condiciones de referencia. ................................ 154 Figura 136. Curva de inactivación frente a dosis UV de los tres organismos analizados.......... 155 Figura 137. Concentración de rotavirus SA-11 empleando la aproximación euleriana............ 156 Figura 138. Molinete Flow Probe 2, empleado para la medición de la velocidad. ................... 157 Figura 139. Localización y denominación de las nueve líneas de medida de velocidad ........... 158 Figura 140. Error relativo entre las medidas de caudal del caudalímetro calibrado y el sistema de medida de caudal por vertedero............................................................................................... 159 Figura 141. Comparación de la velocidad numérico-experimental en la línea 1. ..................... 160 Figura 142. Comparación de la velocidad numérico-experimental en la línea 2. ..................... 161 Figura 143. Comparación de la velocidad numérico-experimental en la línea 3. ..................... 161 Figura 144. Comparación de la velocidad numérico-experimental en la línea 4. ..................... 162 Figura 145. Comparación de la velocidad numérico-experimental en la línea 5. ..................... 162 Figura 146. Comparación de la velocidad numérico-experimental en la línea 6. ..................... 163 Figura 147. Comparación de la velocidad numérico-experimental en la línea 7. ..................... 163 Figura 148. Comparación de la velocidad numérico-experimental en la línea 8. ..................... 164 Figura 149. Comparación de la velocidad numérico-experimental en la línea 9. ..................... 164 Figura 150. Dominio infinitesimal sobre el que se aplica la ecuación de conservación de la intensidad. ................................................................................................................................. 165 Figura 151. Localización del sensor (círculo rojo) en el módulo UV. ........................................ 166 Figura 152. Fotografías del sensor en el módulo UV: a) localización; b) dimensiones. ............ 166 Figura 153. Distancias relativas entre el sensor y las lámparas. ............................................... 167 Figura 154. Comparación numérico-experimental de la intensidad UV en el sensor............... 168 IX Figura 155. Comparación de los histogramas de dosis UV con ambos bancos funcionando (verde), solo el primer banco funcionando (azul) y solo el segundo banco activado (rojo). .... 169 Figura 156. Trayectorias con menos dosis, caso de referencia, solo el banco 1 activo. ........... 170 Figura 157. Trayectorias con menos dosis, caso de referencia, solo el banco 2 activo. ........... 170 Figura 158. Vectores velocidad con (a) y sin (b) placa perforada. ............................................ 171 Figura 159. Vectores velocidad en el plano localizado 100 mm tras la localización que tendría la placa, con (a) y sin (b) placa perforada. .................................................................................... 172 Figura 160. Posición de la nube de partículas y dosis adquirida 25 s tras su liberación para el caso de referencia con (a) y sin (b) placa perforada. ........................................................................ 173 Figura 161. Comparación de los histogramas de dosis UV para el caso el caso de referencia, funcionando con y sin placa perforada. .................................................................................... 173 Figura 162. Campos de velocidad con caudal 112.5 m3/h (a), 225 m3/h (b) y 450 m3/h (c). .. 175 Figura 163. Curvas RTD para distintas condiciones de caudal. ................................................. 175 Figura 164. Distribución de dosis UV en el canal para distintas condiciones de cauldal .......... 176 Figura 165. Intensidad UV bajo distintas transmitancias: 55% (a), 65% (b) y 75% (c). ............. 177 Figura 166. Distribución de dosis UV en el canal para distintas transmitancias ....................... 178 Figura 167. Intensidad UV para las distintas condiciones de fallo de lámpara analizadas. ...... 181 Figura 168. Trayectorias que menos dosis reciben para el caso Q=450 m3/h, T=55%, PUV=143W con solo el primer banco de lámparas activo. .......................................................................... 182 Figura 169. Trayectorias que menos dosis reciben para el caso Q=450 m3/h, T=45%, PUV=143W con solo el primer banco de lámparas activo y la lámpara 1 fundida....................................... 182 Figura 170. Trayectorias que menos dosis reciben para el caso Q=450 m3/h, T=45%, PUV=143W con solo el primer banco de lámparas activo y la lámpara 29 fundida. ................................... 183 Figura 171. Distribución de dosis en el canal bajo condiciones Q=450 m3/h, T=55%, PUV=143W con solo el primer banco de lámparas activo y con alguna de las lámparas averiadas. ........... 183 Figura 172. Intensidad UV en la zona de las lámparas. a) fallo en el sistema de control de la superficie libre; b) funcionamiento sin fallo (T=55%, PUV=143W)............................................. 184 Figura 173. Distribución de dosis en el canal bajo correcto funcionamiento y con fallo en nivel de la superficie libre (Q=450 m3/h, T=45%, PUV=143W, solo el primer banco activo). ............. 185 Figura 174. Trayectorias con menos dosis en el caso Q=450 m3/h, T=55%, PUV=143W con solo el primer banco de lámparas activo, con fallo en el nivel de la superficie libre. .......................... 186 Figura 175. Detalle geométrico de la placa helicoidal. ............................................................. 187 Figura 176. Campo de velocidad en el canal con placa perforada helicoidal (Q=450 m3/h). ... 187 Figura 177. Distribución de dosis en el canal con placa helicoidal (Q=450 m3/h, T=55%, PUV=143W, solo el primer banco activo). .................................................................................. 188 Figura 178. Trayectorias con menos dosis en el caso Q=450 m3/h, T=55%, PUV=143W con solo el primer banco de lámparas activo, con la modificación de la placa perforada. ........................ 189 Figura 179. Detalle geométrico de la modificación del espacio superficie libre-lámpara. ....... 189 Figura 180. Distribución de dosis en el canal bajo la geometría original, la modificación de placa helicoidal y reducción del espacio superficie libre-lámpara ..................................................... 190 Figura 181. Trayectorias con menos dosis para el caso Q=450 m3/h, T=55%, PUV=143W con solo el primer banco de lámparas activo, con la reducción del margen superficie libre-lámparas. 191 XI Índice de tablas Tabla 1. Altura de cada uno de los agitadores sobre el suelo en su posición original. ............... 21 Tabla 2. Concentraciones ASM1 del influente. ........................................................................... 23 Tabla 3. Valor de los coeficientes estequiométricos en el modelo ASM1. ................................. 37 Tabla 4. Ecuación de cada una de las reacciones involucradas en el modelo ASM1. ................. 38 Tabla 5. Porcentaje de volumen con velocidad menor de 10 cm/s bajo las diferentes condiciones de funcionamiento. ..................................................................................................................... 65 Tabla 6. Características de diseño del ensayo de tipo pulso a llevar a cabo en el reactor biológico de la EDAR de San Pedro del Pinatar. ......................................................................................... 75 Tabla 7. Valores de caudal tratado, recirculado y total promedios durante el ensayo RTD del reactor biológico de la EDAR de San Pedro del Pinatar. ............................................................. 76 Tabla 8. Parámetros del ajuste de la curva de Levenspiel a la experimental (modelo 1). .......... 83 Tabla 9. Parámetros del ajuste de la curva de Levenspiel a la experimental (modelo 2). .......... 84 Tabla 10. Parámetros del ajuste de la curva de Levenspiel a la experimental (método 3). ....... 85 Tabla 11. Parámetros de Levenspiel obtenidos por los tres diferentes métodos analíticos y aplicando directamente la ecuación. .......................................................................................... 86 Tabla 12. Parámetros del ajuste de la curva de Levenspiel a los resultados numéricos. ........... 93 Tabla 13. Parámetros RTD obtenidos bajo diferentes condiciones de aireación y caudal. ........ 94 Tabla 14. Porcentaje de volumen con velocidad menor 10 cm/s en cada zona, conf 2. .......... 102 Tabla 15. Porcentaje de volumen con velocidad menor 10 cm/s en cada zona, conf 2 ........... 105 Tabla 16. Volumen estancado en cada zona para cada configuración. .................................... 109 Tabla 17. Parámetros RTD obtenidos con las diferentes configuraciones................................ 111 Tabla 18. Valores de los parámetros RTD en cada régimen de funcionamiento del reactor. .. 123 Tabla 19. Concentraciones biológicas promedio en las dos zonas del reactor obtenidas por los dos métodos de simulación empleados; CFD y simulación de procesos (SProc)...................... 133 Tabla 20. Resumen del funcionamiento del sistema de aireación del reactor biológico. ........ 134 Tabla 21. Caudal de aire promedio, caída de presión y potencia de aireación necesaria en las configuraciones C1 y C2 para cumplir los requerimientos del efluente. .................................. 140 Tabla 22. Caudal de aire promedio, caída de presión y potencia de aireación necesaria en las tres configuraciones analizadas, para cumplir los requerimientos del efluente. ............................ 144 Tabla 23. Niveles de inactivación de los tres organismos considerados en el canal UV para las condiciones de referencia. ........................................................................................................ 155 Tabla 24. Límites máximo y mínimo de caudal registrados en los tres ensayos realizados. .... 159 Tabla 25. Inactivación logarítmica de los organismos analizados en el canal UV con solo el primer banco activado, solo el segundo banco activado y ambos bancos de lámparas conectados. .. 171 Tabla 26. coli bajo las distintas condiciones de operación. ...................................................................... 180 XII Índice de tablas Tabla 31. Consumo específico mínimo de las lámparas UV para conseguir inactivación 3-log de E. coli bajo las distintas condiciones de operación. .................................................................. 180 Tabla 32. Dosis promedio e inactivación bajo distintas condiciones de fallo de lámpara. ....... 181 Tabla 33. Dosis promedio e inactivación bajo condiciones correctas de funcionamiento y con fallo en el nivel de la superficie libre......................................................................................... 185 Tabla 34. Dosis promedio e inactivación bajo la geometría original y la placa helicoidal. ....... 188 Tabla 35. Dosis promedio e inactivación bajo la geometría original y la modificación del margen superficie libre - lámpara .......................................................................................................... 190 1 1. Introducción El campo de investigación de esta Tesis Doctoral es el análisis de instalaciones de depuración de aguas residuales mediante mecánica de fluidos computacional (Computational Fluid Dynamics, CFD), centrándose concretamente en la modelización y simulación de reactores biológicos y canales de desinfección ultravioleta (UV). La investigación trata tanto aspectos de modelización y simulación, como análisis de los fenómenos que ocurren en el interior de las instalaciones. Esta Tesis Doctoral pretende ser una contribución con aportaciones originales al conocimiento que se tiene del tema anteriormente comentado. La búsqueda de la eficiencia del proceso: minimización del gasto energético, pero cumpliendo los requerimientos del efluente, es el objetivo último de este trabajo. 1.1. Origen del estudio Desde finales del siglo XVIII, cuando se produjo la revolución industrial, hasta nuestros días, son muchos los cambios que se han producido en la sociedad. A nivel tecnológico e industrial se han producido avances que se han integrado por completo a la sociedad, lo cual ha contribuido a un mundo globalizado y más cercano. Hoy en día es posible viajar a cualquier parte del mundo en pocas horas, y no solo eso, gracias a los medios de comunicación, internet y la telefonía se puede estar informado de lo que está pasando en cualquier parte del planeta. El camino para llegar a este nivel tecnológico no ha sido sencillo, y se ha pagado un precio por alcanzar esta sociedad. Con el desarrollo industrial se han multiplicado las diferencias entre los países ricos y pobres, aumentando año a año. Desde el punto de vista medioambiental también se ha pagado un precio para alcanzar esta sociedad: palabras como efecto invernadero o cambio climático están a la orden del día en telediarios de medio mundo. A mediados del siglo XX, la parte más crítica de la sociedad industrializada comienza a cuestionarse si los efectos negativos del progreso industrial podrían llegar a ser un problema serio en el futuro, llegando a la conclusión de que se debían de controlar los perjuicios provocados por la industria, era el comienzo del desarrollo sostenible. El desarrollo sostenible fue calando en la sociedad, la gente comenzó a comprender que los recursos que se extraían del planeta no eran inagotables, y que el medio natural no tenía capacidad infinita para absorber la contaminación. En los años 80 se comprendió definitivamente que era necesario regular la emisión de contaminantes, así como el consumo de recursos. Hoy en día el cuidado del medio ambiente es una realidad, y la sociedad es consciente del problema de los residuos. La sociedad genera residuos tanto sólidos, líquidos y gaseosos, y está claro que hay que eliminarlos o tratarlos, pues el medio natural no tiene capacidad para hacerlo, y será la propia sociedad que los genera la que sufrirá los efectos negativos de no tratar correctamente los residuos. El tratamiento de aguas residuales tiene como propósito tratar el agua residual procedente de la industria o la población, para evitar que al devolverla al medio natural receptor (lago, río, etc.) produzca efectos negativos sobre él. Algunos de los efectos que produce el agua residual no tratada sobre el medio natural son los siguientes: 2 1. Introducción − Tapiza la vegetación de las riberas con residuos sólidos gruesos que lleva el agua residual, tales como plásticos, utensilios, restos de alimentos, etc. − Acumulación de sólidos en suspensión sedimentables en fondo y orillas del cauce, tales como arenas y materia orgánica. − Consumo del oxígeno disuelto que tiene el cauce por descomposición de la materia orgánica y compuestos amoniacales del agua residual. − Formación de malos olores por agotamiento del oxígeno disuelto del cauce que no es capaz de recuperarse. − Entrada en el cauce de grandes cantidades de microorganismos entre los que pueden haber elevado número de patógenos. − Contaminación por compuestos químicos tóxicos o inhibidores de otros seres vivos (dependiendo de los vertidos industriales) − Aumenta la eutrofización al portar grandes cantidades de fósforo y nitrógeno. Por otro lado, la escasez de precipitaciones se ha convertido en un factor limitante del desarrollo socioeconómico de muchas regiones del mundo, como es el caso del sur de España. Aunque hay discrepancias sobre las causas de este periodo de sequía: algunos lo achacan al cambio climático, otros afirman que son fenómenos cíclicos y otros que se debe a una combinación de los dos anteriores; lo cierto es que la falta de agua produce problemas económicos, políticos y sociales. Es un problema que llega incluso a ser tema de Estado, existiendo diferentes organismos y entidades públicas destinadas a velar por la correcta y justa gestión del agua. Este problema afecta con mayor gravedad a la Región de Murcia, especialmente en sectores de gran peso económico como la agricultura y el turismo. Ya en el Plan Estratégico de Desarrollo de la Región de Murcia para el periodo 2000-2006 se subrayaba la racionalización y optimización del uso del agua como línea prioritaria para el crecimiento de la región. Dentro de esta optimización, la reutilización de las aguas residuales depuradas como fuente de agua adicional cobra especial importancia, ya que permite aliviar las necesidades hidrológicas de la agricultura, disminuyendo la presión ejercida sobre los acuíferos y permitiendo su recuperación. Más recientemente, en la elaboración de la Estrategia de Investigación e Innovación para la Especialización Inteligente de la Región de Murcia (RIS3Mur), que tiene como objetivo identificar y priorizar las actividades en las que la Región presenta un mayor potencial de competitividad y desarrollo, y dentro del Plan “Europa 2020”, exigido para acceder a los fondos estructurales 2014-2020, se señalan las actividades vinculadas al ciclo del agua (tratamiento, depuración y gestión) como campo prioritario de investigación e innovación, destacando la importancia de dicho campo en el sector agroalimentario de la Región. Dentro de este contexto, queda clara la importancia del tratamiento de aguas residuales en la Región de Murcia, así como la necesidad de una depuración eficiente, que asegure que el agua depurada cumpla los requisitos necesarios para su reutilización en la agricultura (o para verterla al medio receptor), procurando tener el menor gasto energético posible. Las estaciones depuradoras de agua residual (EDAR) son las encargadas de realizar el tratamiento de aguas residuales. Las EDAR se pueden clasificar en dos tipos principales: − EDAR urbanas, que reciben el agua residual de una aglomeración urbana. − EDAR industriales, que reciben el agua residual de una o varias industrias. 3 Figura 1. Vista general de la EDAR de Aspe (Alicante), ejemplo de EDAR urbana. El tratamiento depende de varios factores, principalmente de la procedencia del agua residual, pero en líneas generales, los tratamientos convencionales que se realizan en una EDAR son los siguientes. − Pretratamiento: Consiste en la eliminación por tratamientos físicos de los sólidos y grasas presentes, para evitar que dañen los equipos de tratamiento posteriores. Los principales tratamientos en esta etapa son: • Desbaste, para la eliminación de gruesos no solubles como plásticos, envases. • Desarenado, para la eliminación de sólidos de pequeño tamaño. • Desengrasado, para la eliminación de sólidos y líquidos no miscibles de menor densidad que el agua. − Tratamiento primario: Consiste en una decantación para eliminar las partículas de menor tamaño que no han podido ser eliminadas en el pretratamiento. − Tratamiento secundario: Se trata de un tratamiento biológico, con la ayuda de bacterias y mediante la adición de oxígeno se consigue oxidar la materia orgánica presente en el agua. Existen varios tipos de tratamientos secundarios (fangos activos, lechos bacterianos, reactores biológicos de membrana), pero el principio de tratamiento es común. La oxidación biológica de la materia orgánica presente en el agua residual es el proceso clave en la depuración. Tras el tratamiento biológico, se separa el agua depurada de los fangos, tradicionalmente dicha separación se lleva a cabo mediante un proceso de decantación. Sin embargo, en la actualidad se están imponiendo paulatinamente las tecnologías basadas en la filtración para la separación fango-agua. Un ejemplo de esta tecnología son los reactores biológicos de membrana (MBR). Su principal ventaja es que necesitan mucha menos superficie que un decantador convencional para separar el mismo caudal de agua. Por otro lado, necesitan un mantenimiento mayor para evitar la obstrucción de los poros. 4 1. Introducción − Tratamiento terciario: En esta parte se engloban los tratamientos físico-químicos destinados a tratar alguna de las características del agua tratada, como eliminar la presencia de bacterias, virus y gérmenes. Aunque el proceso clave de la depuración es el tratamiento secundario, la desinfección (englobada dentro del tratamiento terciario) del agua cobra un papel esencial cuando el agua depurada es utilizada para irrigación. Ya que es importante eliminar bacterias como la Escherichia Coli (E. Coli), para que no se transmita a los cultivos. Uno de los tratamientos terciarios más utilizados es la desinfección ultravioleta (UV), que consiste hacer pasar el agua residual entre lámparas que irradian luz ultravioleta, la cual tiene la capacidad de inhibir la capacidad reproductora de los organismos presentes en el agua. Además de la línea de agua descrita, las EDAR cuentan con la llamada línea de fangos, que consiste en el tratamiento de los concentrados extraídos del agua depurada. Los fangos reciben un tratamiento de espesamiento, digestión y deshidratación, para convertirlos en residuos susceptibles de ser tratados como urbanos o incinerados, aunque también se pueden obtener productos de compostaje para la agricultura, o biogás que puede usarse como combustible. La figura 2 muestra la línea de proceso de una depuradora convencional. Figura 2. Línea de proceso de una EDAR convencional. Fuente: Wikipedia. La depuración de aguas residuales está regulada por la directiva europea 91/271/CEE que establece, entre otras imposiciones, los requerimientos del efluente. El no cumplimiento de esta directiva acarrea el pago de multas e indemnizaciones por parte del explotador, controlador, propietario de la EDAR o incluso el mismo Estado español en función de la falta. En 2016, España contaba con más de 2000 estaciones de depuración, que trataron un total de 4100 hm3 (unos 90 m3/ habitante). Por otro lado, en el tratamiento de aguas residuales se ven involucrados, entre otros, procesos de bombeo, aireación y agitación que suponen un alto coste energético. Por motivos económicos, y sobre todo ambientales, queda clara la necesidad de mejorar la eficiencia de las EDAR; cumplir los requerimientos del efluente, tanto para verterlo al medio natural como para reutilizarlo en irrigación, pero con el menor coste energético posible. Para tal fin, es necesario abordar una rigurosa investigación de las distintas instalaciones de depuración. El estudio de los procesos y plantas de depuración engloba varias disciplinas: química y biología tienen un papel importante en el estudio de las reacciones que ocurren en los reactores, así como en el análisis del agua. Sin embargo, el papel de la Mecánica de Fluidos es fundamental en el estudio. El análisis fluidodinámico del flujo proporciona importante información del proceso de depuración, permitiendo así mejorar el funcionamiento fluidodinámico de las plantas. Durante los últimos años, el Departamento de Ingeniería Térmica y de Fluidos de la Universidad 5 Politécnica de Cartagena (UPCT) ha desarrollado, junto con la Entidad de Saneamiento y Depuración de Aguas Residuales de la Región de Murcia (ESAMUR), distintos trabajos de investigación con la finalidad de mejorar la fluidodinámica de las instalaciones de las Estaciones de Depuración de Aguas Residuales (EDAR) pertenecientes a la Comunidad Autónoma de Murcia. Esta colaboración realizada pone de manifiesto dos aspectos que justifican esta Tesis Doctoral: por un lado la necesidad de analizar y mejorar la fluidodinámica de las instalaciones de las EDAR, por otro lado la adecuación de la herramienta CFD para abordar dichos análisis. La CFD resulta una herramienta ideal para estudiar la fluidodinámica de las instalaciones que componen las estaciones de depuración, debido a su capacidad para abordar cualquier tipo de problema de fluidos, así como la facilidad para obtener abundante información del problema. 1.2. Estado del arte Los métodos de evacuación y depuración de aguas residuales se remontan a la antigüedad. Se han encontrado instalaciones de alcantarillado en lugares prehistóricos de Creta y en antiguas ciudades asirias. Los romanos ya usaban alcantarillados para drenar el agua de sus ciudades. Varios siglos después, en el siglo XIX algunas ciudades como Londres comenzaron a construir canales en sus calles, que utilizaban como desagües para eliminar los desechos humanos, que finalmente desembocaban al río. A finales del siglo XIX, se empezó a reconocer que el vertido directo de los deshechos en el río provocaba problemas sanitarios, tales como fiebres tifoideas. En 1881 Robert Koch realiza el descubrimiento más importante de la historia del tratamiento de aguas, al descubrir la capacidad del cloro para matar las bacterias del agua. A principios del siglo XX algunas ciudades norteamericanas comienzan a implantar sistemas de tratamiento para el agua que abastecía a la población; practica que durante la primera mitad del siglo XX se expandió por toda Europa. Así, en España, a mediados del siglo XX el agua potable llegaba a más del 90% de la población, pero el uso de plantas depuradoras de aguas residuales estaba restringido a muy pocas ciudades. Fue en los años 80, cuando la población comenzó a concienciarse de la necesidad de un “desarrollo sostenible”, que asegurase el abastecimiento de agua de calidad a las generaciones futuras. Bajo dicho contexto, las administraciones comenzaron a regular la depuración de las aguas residuales. Tal y como se ha comentado anteriormente, actualmente la directiva europea 91/271/CEE obliga, salvo condiciones excepcionales, a tratar el agua residual procedente de todas las aglomeraciones urbanas de más de 2000 habitantes, lo que en España supone más de 2000 EDAR en funcionamiento. Las fases que componen el tratamiento convencional de aguas residuales (pretratamiento, tratamiento primario, secundario y terciario) siguen siendo las mismas y en el mismo orden que en el siglo pasado. Sin embargo, los equipos que se encargan de hacer cada uno de los procesos sí van evolucionando y cambiando con el tiempo. Para el diseño de una EDAR se parte de los parámetros de entrada del agua de entrada (caudal, carga biológica (DQO), nitrógeno, sólidos en suspensión, fósforo, coloiformes totales, etc.) y de los deseables a la salida (especificados por la normativa de cada Comunidad Autónoma), otros condicionantes como temas ambientales, económicos o de espacio disponible en el terreno también han de ser tenidos en cuenta. Tradicionalmente, el diseño de las distintas instalaciones involucradas en el proceso de depuración se realiza en base a manuales (ej. Spellman 2013), que sirven para dimensionar el reactor, en función de los parámetros de entrada y salida. Sin embargo, en la mayoría de los casos, estos manuales solo sirven como guía, y el diseñador tiene que usar su experiencia para solventar algunos problemas particulares de su planta, como puede ser la eliminación de algún contaminante específico o el uso de una tecnología diferente a la considerada en el manual. 6 1. Introducción Otro problema de los manuales es que no tienen en cuenta la fluidodinámica en el interior de los reactores y canales involucrados en el proceso; en la mayoría de los casos suponen un patrón de flujo ideal (flujo pistón o flujo perfectamente mezclado) y en base al caudal y al tiempo de contacto necesario seleccionan las dimensiones del equipo. Sin embargo, la eficiencia de la depuración depende en gran medida de los fenómenos fluidodinámicos que se producen en el interior de los equipos (Vilker 1993, Ouedraogo et al. 2016), deficiencias fluidodinámicas tales como zonas muertas, cortocircuitos o recirculaciones pueden llegar a causar una incorrecta depuración del agua residual, por lo que han de ser evitadas mediante un correcto diseño fluidodinámico. Desafortunadamente, predecir el comportamiento fluidodinámico en el interior de una instalación de depuración presenta diferentes dificultades. Las ecuaciones que rigen el comportamiento del fluido son las ecuaciones de Navier-Stokes, consistentes en los balances de masa y cantidad de movimientos, que proporcionan como solución los campos de velocidad y presión. A pesar de ser publicadas en el siglo XIX, aún hoy en día, las ecuaciones de Navier-Stokes no tienen solución analítica conocida, tan solo es posible resolver simplificaciones de las mismas (flujo unidimensional laminar, flujo unidimensional ideal, fluidoestática, etc.), insuficientes en la mayoría de ocasiones para predecir la fluidodinámica en el interior de las instalaciones de EDAR. Otra vía para el análisis de la fluidodinámica en el interior de las instalaciones de depuración es la experimentación: mediante el empleo de sensores, se obtienen valores de velocidad y presión en distintas localizaciones del reactor o canal. Sin embargo, el estudio experimental de las instalaciones de depuración presenta algunas limitaciones: − Universalidad de los resultados: los valores de velocidad han sido obtenidos para una instalación bajo unas condiciones de funcionamiento determinadas, no aseguran la predicción del flujo bajo distintas condiciones de funcionamiento o geometría. − Detalle de la información: los sensores proporcionan valores de velocidad en localizaciones específicas de la instalación, no proporcionan el campo completo de velocidades. El precio de los sensores es un limitante a la hora de seleccionar el número de medidas espaciales a registrar. − Naturaleza del agua residual: el agua residual contiene sólidos en suspensión, lo que imposibilita el uso de sensores ópticos y dificulta el uso de sensores basados en el movimiento mecánico, ya que se pueden obstruir. Por otro lado, las velocidades del agua en los canales y reactores de depuración son del orden de cm/s, lo que supone una limitación más al aparato de medida, ya que ha de ser capaz de detectar velocidades del agua bajas. En este contexto, ante las limitaciones que presentan los métodos analíticos y experimentales para el análisis del flujo, la Mecánica de Fluidos Computacional (CFD), surge como una herramienta interesante para el análisis del flujo en instalaciones de depuración. Aunque se entrará en detalle más adelante, la CFD es una herramienta cuyo fin es aproximar la solución de las ecuaciones de Navier-Stokes mediante métodos matemáticos. Con ello, logra solventar las limitaciones que tenían los métodos analíticos y experimentales, ya que consiguen proporcionar en detalle los campos de velocidad y presión, para las diferentes geometrías y condiciones de funcionamiento. Por otro lado, es necesaria la validación experimental del modelo, con el fin de asegurar la validez de sus resultados. A finales del siglo pasado, la CFD comenzó a usarse como herramienta para el diseño de las EDAR, en combinación con el estudio experimental. En los últimos años su uso en el sector se ha extendido, consolidándose como una herramienta útil y eficaz para el análisis fluidodinámico. Su potencial no se reduce a la fluidodinámica, sino que es capaz de modelizar y simular fenómenos de diferente naturaleza tales como bioquímica o radiación. A continuación se detalla el estado del arte en cuanto al empleo de CFD para el diseño y análisis de reactores biológicos y sistemas de desinfección ultravioleta (UV) en EDAR.	40,839
https://openalex.org/W1574283480_1	Spanish-Science-Pile	Open Science	Various open science	2,014	Brasil y Colombia, antes y después de la crisis económica mundial de 1929.	None	Spanish	Spoken	7,506	12,326	ESTRATEGIAS COMERCIALES DE LOS PRINCIPALES PRODUCTORES DE CAFÉ. Brasil y Colombia, antes y después de la crisis económica mundial de 1929.* Por: Carlos Alberto Murgueitio Manrique** Resumen Este artículo pretende analizar las estrategias económicas y comerciales empleadas por los principales productores mundiales de café para impedir el desplome de los precios del grano durante la década de 1930, caracterizada por el colapso coyuntural del sistema financiero a nivel mundial y la crisis en los precios de las manufacturas y las materias primas en los mercados internacionales. Además, busca mostrar las coincidencias y las distancias entre Brasil y Colombia en referencia a la puesta en marcha de medidas económicas proteccionistas o estatistas, en boga durante la época, analizando los conflictos internos entre los gobiernos y las agremiaciones cafeteras. Al mismo tiempo, este artículo busca advertir acerca de la iniciativa binacional que emergió como alternativa a la tradicional competencia, encaminada a la consolidación de un cartel cafetero que pudiese, a partir de mecanismos económicos conjuntos, incidir en el mantenimiento de los precios del grano en el mercado internacional durante la década de los treinta. Palabras claves * Artículo de Investigación Científica tipo 2: de reflexión, según clasificación de Conciencias. ** Politólogo – Universidad de los Andes, Bogotá Candidato a Magíster en Historia Contemporánea de América – Universidad Central de Venezuela, Caracas Profesor – Universidad del Valle – Universidad Javeriana - Cali 1 Defensa de los precios del café, crisis internacional del capitalismo, República Liberal, Getulio Vargas, pacto internacional del café, Congresos Panamericanos del café. Abstract This article analyses the economical strategies employed by the principal world coffee producers to impede the collapsing of the grain prices during the thirties, period characterized with the international financial crash that resulted in a general crisis of the international markets including manufactures and agricultural sectors. Besides, it searches to expose the coincidences and distances between Brazil and Colombia in reference to the development of protectionist policies, popular during this age, analysing the internal conflicts between governments and coffee trade unions. At the same time, this article seek to warn about the binational initiatives that emerged as an alternative to the traditional competition, searching to consolidate a coffee cartel that could manoeuvre common policies in order to fortify the maintenance of the prices of the grain in the international markets during the thirties. Key words Liberal republic, Getulio Vargas, international crisis of the capitalism, economic strategies for the defense of the prices of the coffee, international pact of the coffee, competence among the main producers, Panamerican congress of coffe. 2 La crisis de la economía cafetalera en el Brasil (1889 – 1934) A finales del siglo XIX, Brasil se había convertido en el principal productor mundial de café. Cerca del 70% del grano existente en el mercado internacional provenía de este país, y el apoyo recibido de parte de los estados productores y de las instituciones federales respaldaban el crecimiento de los cultivos a partir de la expansión de la colonización hacia nuevas fronteras agrícolas ubicadas al norte del estado de Sao Paulo. Además, en el último decenio del siglo XIX se había generado una situación excepcionalmente favorable para la expansión del cultivo cafetalero en el Brasil, ya que la oferta del grano no brasileño atravesaba una etapa de dificultades, siendo perjudicada por las enfermedades casi la totalidad de la producción asiática. La descentralización impuesta por la república desde 1889 les había dotado, sobre todo a los estados ricos un autonomía considerable en relación a sus políticas productivas, de esta manera Sao Paulo extendía su mercado fomentando la inmigración en masa de obreros agrícolas europeos, indispensables para reemplazar a la mano de obra negra tradicional, liberada de la esclavitud un año antes del triunfo republicano. Los efectos estimulantes de la gran inflación del crédito de ese periodo benefició de dos maneras a la clase propietaria cafetalera. Tal y como lo dice Celso Furtado; “(...), proporcionó el crédito suficiente para financiar la apertura de nuevas tierras y elevó los precios del producto en moneda nacional a partir de la depreciación cambiaria. La producción brasileña que había aumentado de 3.7 millones de sacos (de 60 kg.), en 1881 a 5.5 millones en 189, alcanzando en 1901 los 16.3 millones de sacos”1. 1 Furtado, Celso. Formación económica del Brasil, Fondo de Cultura Económica, México, 1962. (pp. 181). 3 La elasticidad de la mano de obra y la abundancia de tierras en los países productores del trópico, eran claros indicios de los posibles descensos coyunturales y estructurales en los precios de los granos, además, estos pronósticos se materializarían más mientras se siguiesen construyendo ferrocarriles, puertos fluviales y marítimos que permitirían una mejora en la distribución y comercialización de los productos. En la medida que los mercados internacionales se saturaran de artículos de las economías coloniales o semicoloniales, los problemas económicos se tornarían inviables para los productores. Estos no tenían mucha libertad para escoger el producto al cual destinarían sus inversiones, se veían forzados por las presiones de la demanda y los caprichos de los precios fluctuantes de las materias primas. Todas estas variables hacían inevitable que la oferta de café tendiese a crecer, tanto en el Brasil como en los demás países potencialmente productores. La ventaja relativa del café frente a otros productos tropicales se mantendría mientras la oferta pudiese cubrir el mercado mundial, pero de presentarse una saturación, el precio descendería vertiginosamente y para superar la posible crisis, el Estado brasileño tendría que disponer de sus reservas financieras para responder en defensa de las clases terratenientes y de la estabilidad o rentabilidad en los precios. La primera crisis se presentó en 1893, en la cual comenzaron a declinar los precios del grano en el mercado mundial, el valor medio del saco exportado en aquel año fue de 4.09 libras esterlinas, descendiendo hasta 2.91 libras en 1896 y 1.48 libras en 18992. La prolongación de la crisis en el precio del grano impidió que la devaluación amortiguara su estabilidad provocando la intervención del mercado por medio de las políticas centrales del Estado. La superproducción, palpable en las existencias acumuladas durante esos años pesaban sobre los precios, provocando una pérdida 4 permanente en el ingreso de los productores y por ende una intranquilidad social aguda. La idea de retirar parte de la producción tomó fuerza gracias al apoyo del poder político y financiero de los dirigentes de los estados cafetaleros. El monopolio del Brasil en el mercado mundial le permitía a sus autoridades jugar un papel decisivo en el negocio, la base para explotar ese potencial era aumentar los ingresos elevando los precios del grano en el mercado. En 1905 durante las reuniones del Comité de Finanzas del Senado, se presionó al gobierno de turno para que abandonara los postulados económicos del modelo liberal clásico y el razonamiento basado en las leyes del mercado y adoptara mecanismos proteccionistas e interventores que aseguraran la estabilidad de los sectores productivos, especialmente aquellos vinculados con la agroindustria exportadora. Los argumentos para llevar a cabo estas políticas estaban respaldados en el papel primordial que jugaba la producción brasileña en el mundo, tal y como lo exponen las siguientes citas, “(...), Brasil está en una posición excepcional y no debe aceptar simplemente los precios del mercado como algo dado sino que debe fijarlos reteniendo café de exportación”3, “(...), Brasil no tiene competidores ni tendrá alguno en el mundo, Brasil debe ser libre de capitalizar su posición elevando el precio internacional del café”4. La descentralización republicana había reforzado el poder de los caficultores a nivel regional. El Acuerdo de Taubaté, (1906), sería la manifestación real de los logros políticos de la oligarquía productora, que buscó trasladar sus costos económicos al Estado a través de una política de valorización del café que consistía en el desarrollo de cuatro puntos: a) la 2 Los precios referidos fueron extraídos de: Furtado, Celso. Ibídem, (pp. 182). Pereira Reis, María de Conceisao. The Agrarian roots of Authoritarian Modernization in Brasil, 1880 – 1930, Tesis Doctoral, Massachuset Institute of Tecnology, 1979. (pp. 180). 3 5 intervención en el marcado para comprar los excedentes y restablecer el equilibrio entre la oferta y la demanda, b) la financiación de las compras con empréstitos extranjeros, c) el pago de las obligaciones con el sistema financiero internacional por medio de los impuestos en oro sobre cada saco de café exportado y d) la adopción de una política restrictiva a nuevos productores con el fin de entorpecer la expansión de futuras plantaciones. Dentro de la estructura federal cada estado intentaba utilizar su poder a nivel nacional para lograr sus objetivos políticos. En el caso de Sao Paulo, eso significaba asegurar políticas que favorecieran a los productores de café, pues para 1920 el 12% de las plantaciones más grandes de Sao Paulo controlaban dos terceras partes del potencial productivo de la industria. Sin embargo, Sao Paulo solo no podía controlar al legislativo, obligándolo a tejer alianzas con intereses de otras regiones exportadoras de productos primarios al mercado internacional. “Para lograr sus objetivos, los estados hacían esfuerzos para volver a los presidentes dependientes, entregándoles o retirándoles grandes lotes de votos en las elecciones presidenciales o con una delegación grande y unificada en el Congreso, capaz de usar su poder de voto para aprobar las políticas deseables y bloquear las indeseables”5. En su esfuerzo por controlar los resultados legislativos, la delegación de Sao Paulo mantuvo estrechas relaciones de alianzas con las de Minas Gerais y Río Grande do Sul. Pese a los éxitos logrados por los sectores productores dominantes, las relaciones entre los grupos que empezaban a ejercer presión sobre el gobierno se complejizaron. La 4 Krasner, Stephen. The politics of primary commodities. A study of the coffee, (1900 – 1970), Harvard University, 1971. (pp. 119). 5 Bates, Robert. Política Internacional y Economía abierta, la economía política del comercio mundial del café, Tercer Mundo Editores, Bogotá, 1999. (pp. 39). 6 depreciación de la moneda trajo beneficios para el sector exportador de café y para los productores de otras exportaciones, como los azucareros del nordeste, los exportadores de algodón del norte y del centro, la industria cauchera del Amazonas, los cultivadores de cacao y los explotadores de minerales. Pero, la importancia creciente de la clase media urbana, dentro de la cual se destacaban la burocracia civil y militar, los comerciantes importadores e industriales, imponían la adopción de nuevas formas de estrategias económicas. Si se quería captar sus votos para el juego político era indispensable la revisión de las medidas de depreciación cambiaria y su combinación con políticas de bienestar, las cuales no podrían generarse si los capitales del Estado seguían siendo invertidos en la defensa de los precios del café. En el período entre 1923 – 1929, los gastos en capital público se incrementaron abruptamente en términos relativos y absolutos a expensas de las obras públicas. Los productores y exportadores de bienes primarios no solo buscaban la devaluación del milreis, y un mayor protagonismo del Estado en la economía como elemento que asumiría parte de las pérdidas ocasionadas por el descenso de los precios del café, también exigían una protección arancelaria mayor para aumentar el nivel de sus ventas en el mercado interno, el fomento encaminado a la construcción de carreteras y la protección a la agricultura. El objetivo era reducir los costos del transporte que gracias a la construcción de ferrocarriles que unieran a las zonas productivas del interior con los puertos, aprovechando que la mayor concentración de la población estaba ubicada en los ríos y las costas. La apelación de los caficultores a los empréstitos internacionales les permitió gozar de combustible necesario para mantener estas políticas de estabilización de los precios hasta entrados los años veinte. En 1921, Epistacio Pessoa, presidente de Brasil, adoptó 7 ciertos postulados económicos convenientes para los productores de café, estas medidas se conocieron durante la época bajo el lema de la “Defensa Permanente del café”. Según él, al ser el café la fuente de la mayor parte de las exportaciones de Brasil, su defensa constituía un problema nacional, cuya solución exigía al gobierno fijar políticas económicas y financieras sanas. Las ganancias de café eran invertidas en la creación de la industria urbana, por medio de la participación de capitales en dichas empresas o colocando los ahorros en el mercado de capitales, traducidos en créditos industriales”6. El éxito del plan elaborado por los sectores caficultores, mantuvo una tendencia firme en los precios y ganancias elevadas que hacían atractivo el negocio y por lo tanto una continuidad en el crecimiento de plantaciones. La medidas tomadas en tal sentido no fueron fructíferas, hubiese sido necesario que se ofreciesen a los empresarios agroindustriales otras oportunidades igualmente lucrativas, pero la comodidad brindada por la industria del café los incapacitó de hacerlo. … el mecanismo de defensa de la economía cafetalera era, en última instancia, un proceso de transferencia hacia el futuro de la solución de un problema que se tornaría cada vez más grave”7. La producción de café, causa de los estímulos artificiales creció fuertemente en la mitad de los años veinte. Entre 1925 y 1929 tal crecimiento fue de casi un ciento por 6 7 Prado Junior, Ciao. A revolucao Brasileira, Editora Brasiliense, Río de Janeiro, 1966. (pp. 257 – 269). Furtado, Celso. Ibídem, (pp. 184). 8 ciento, lo que revela una cantidad exorbitante de arbustos sembrados desde el Acuerdo de Taubaté. En 1926, el ex – gobernador de Sao Paulo Washington Luís, toma posesión del cargo de presidente del gobierno federal de la república. Para apoyar las medidas coherentes con la defensa del café, crea la Caixa de Estabilizacao y el Banco del Estado de Sao Paulo para intervenir en el mercado de divisas permitiéndole al Instituto del Café de Sao Paulo, a partir de una coordinación de funciones y obligaciones liberarse de parte de sus compromisos. Estas instituciones estarían en la obligación de comprar y almacenar el grano, que provocarían el aumento del precio en dólares del café y por ende el aumento de los ingresos de divisas del Brasil, además, del control de los embarques de café en el puerto de Santos. A medida que la producción se incrementó en respuesta a las nuevas políticas de precios, los inventarios aumentaron y también se retrasaron los embarques”8. La circularidad de los factores llevó a Brasil a la sobresaturación de su oferta. La retención del grano hacía el mantenimiento de precios rentables en el mercado internacional. Esos precios elevados, impulsaron mayores inversiones de capital en las plantaciones mientras la demanda continuaba evolucionando dentro de las líneas tradicionales de su comportamiento. Si se contraía un poco en las depresiones, también se expandían poco en las etapas de gran prosperidad. No obstante, la gran elevación del ingreso real en los países industrializados de Europa y los Estados Unidos en el decenio de los veintes, ese superávit no modificaría en nada la demanda de café. 9 En los Estados Unidos, principal importador, donde el ingreso real per capita creció cerca del 35% en el transcurso de ese decenio, el consumo de café se mantuvo en torno a las 12 libras – peso por habitante, aunque los precios al por menor se mantuviesen estables“9. El superávit de las existencias acumuladas entre 1927 – 1929 alcanzó la apreciable suma de 4.2 millones de contos, o sea cerca de 24 mil millones de cruceiros, sobrepasando en un 10% al producto bruto brasileño del último años antes de que se desatara la crisis internacional del capitalismo y las transformaciones propias de la revolución militar de 1930. Pese a las medidas de contención, empleadas para mitigar los efectos de la sobreproducción, esta alcanzó sus niveles más altos en 1933, como efecto de las nuevas plantaciones realizadas entre 1928 – 1929. La profunda depresión en la que se encontraba el mercado internacional de capitales impedía que los productores o el mismo Estado contaran con préstamos debido a que el crédito había desaparecido y además si este hubiese podido obtenerse, Brasil ya no contaba con el respaldo suficiente. Sus reservas se habían evaporado por las políticas de defensa permanente del café. La gran acumulación de existencias, la rápida liquidación de las reservas metálicas y las precarias perspectivas de financiación de las grandes cosechas previstas para el futuro, aceleraron la caída del precio internacional del café iniciada conjuntamente con las de los demás productos primarios a finales de 1929. 10 La revolución de Vargas en 1930, había recibido el respaldo del ejército, que en su mayor parte estaba acantonado en el estado de Río Grande do Sul, en la frontera sur del 8 Bates, Robert. Ibídem, (pp. 63). Ver: Capacidad de los Estados Unidos para absorber los productos latinoamericanos, CEPAL, 1951. 10 Furtado, Celso. Ibídem, (190 - 191). 9 1 país. Este estado estaba caracterizado por su enorme producción de alimentos, tanto carne como arroz, que se vendían en el mercado internacional, además de haberse convertido en una zona productora de manufacturas sustitutas a las importaciones. El levantamiento militar había surgido por la presión provocada por la depreciación del milreis que acompañó la intervención del mercado modificando los precios relativos de tal forma, que redujo el valor real de la moneda y de los ingresos militares. Vargas exigió que los cultivadores asumieran una proporción mayor de las pérdidas o los costos. Su gobierno aumentó los impuestos a las exportaciones, especialmente del café y obligó a los agricultores a que entregaran una parte de las cosechas para el almacenamiento y la distribución. También trasladó una porción mayor de costos a los nuevos competidores que intentaban explotar y con ello debilitar su programa, para tal fin impulsó un gravamen impositivo y la prohibición de nuevas siembras. La vigilancia sobre el café se ejercía hasta el momento en Sao Paulo a través del Instituto del café de ese estado, “(...), desde mayo de 1931estas funciones pasaron al Consejo Nacional del Café”11, creado por el gobierno federal. Frente a las dificultades externas de un mundo en crisis económica y financiera, se emplearon estímulos para el sector interno afectado por el déficit presupuestario, con las consiguientes barreras a las importaciones, las medidas cambiarias impulsadoras de la producción local y la sustitución de importaciones. La defensa de la industrialización no era una corriente explícita, pero ya se seguía y poco después recibió un gran apoyo. Para la compra de stocks o existencias se crearon varios impuestos especiales y se conservaron los ya existentes. Se ordenó la destrucción de un millón de sacos anuales durante por lo menos 11 Iglesias, Francisco. Breve historia contemporánea de Brasil, Fondo de Cultura Económica, México, 1983. P.66 1 12 años. Inmensas cantidades de café se quemaron o arrojaron al mar, lo que en el país y fuera de él causó temor y provocó protestas, este procedimiento se seguiría practicando hasta 1944. La brusca caída de los precios a nivel mundial y la falla del sistema de convertibilidad de las monedas por los cambios en el patrón oro, ocasionaron la caída del valor externo de la moneda brasileña. “(...), Esa caída asumió proporciones catastróficas, pues de septiembre de 1929 a ese mismo mes de 1931, la baja fue de 22.5 centavos de dólar por libra a 8 centavos”12. La mayor parte de las pérdidas sería entonces transferida hacia el conjunto de la colectividad, a través del alza en los precios de las importaciones. A pesar de estas bajas en los precios, la verdad era que el mercado internacional no podría absorber el total de la producción, la demanda era poco elástica, en función de los precios y la sobreproducción era tan enorme que hacia imposible la restauración del equilibrio. Haciendo a un lado la preocupación de defender los precios, se abría la posibilidad de forzar al mercado, lográndose un aumento en el volumen físico exportador en un 25% entre 1929 y 1931. Aun así, una apreciable parte de la producción quedaba almacenada y sin ninguna posibilidad de colocarse en el mercado. Era obligatorio imponer medidas suplementarias. La producción prevista para los diez años siguientes excedía sobradamente la capacidad previsible de absorción de los mercados compradores, la destrucción de una parte de la cosecha se impuso como única solución a la problemática. De esta manera se rompió el círculo y se impidió que el precio del café siguiera desplomándose llegando con 1 el tiempo a la armonización entre la oferta y la demanda y a un incremento en los precios del grano. La decisión de quemar los excedentes de la producción en grandes cantidades para que no llegaran a incidir en el mercado no tuvo impacto en el mantenimiento de los precios ni en Nueva York ni en Londres, pero sí fomentó la industrialización, poco apreciada en aquel momento por los sectores económicos relevantes. El resultado de mantener la demanda interna combinada con una progresiva devaluación de la moneda con tal de mantener en declive a las importaciones resultó ser un estimulante para la industria básica de Brasil. Esta espontánea industrialización sería ayudada por una política intervencionista conciente, fomentada por el Estado Novo y el repudio de Vargas hacia las políticas de sino liberal. El intervensionismo produjo el anhelado desvío de Brasil de las corrientes doctrinarias de la economía clásica o del liberalismo manchesteriano. Pese a las reformas en el tratamiento de la cuestión cafetera desde el ascenso al poder de Vargas y el establecimiento de la Nueva República, el precio del café atravesó todo el decenio de los treintas inmune a los favorables vientos de recuperación desde 1934. Después de alcanzar su punto más bajo en 1933, la cotización internacional del grano se mantuvo sin variación hasta 1937, para luego descender aun más en los últimos dos años del decenio. Si lo comparamos con los precios de otros productos primarios, el café mantuvo precios desfavorables por más tiempo que todos los demás. Por ejemplo, el precio del azúcar, subió en 140% entre 1933 – 1937; el cobre se elevó un poco más de un 100%, mientras el precio del café en 1937, era igual al de 1934 e inferior al de 193213. 12 13 Ver: Burns, Bradford. A history of Brasil, Columbia University Press, New York, 1993. (pp. 313 – 380). Ver: Furtado, Celso. Ibídem, (pp. 193). 1 Para el primer semestre de 1937, era visible la disminución de los stocks de té, azúcar, trigo seda y caucho, productos que alcanzaron los niveles más bajos en reservas desde 1930, mientras el algodón sólo registró cifras inferiores en 1935. Por su parte, el café las registró superiores, en el semestre en referencia, a las de todos los años anteriores”14. A partir del período 1929 – 1930 no hay un año que no muestre sobrantes considerables en la producción. La cosecha de 1933 – 1934 arrojó uno de más de 15 millones de sacos y la de 1936 – 1937, más de 13 millones. Las anormales cosechas brasileñas restaron mucho de su eficacia a la política de destrucción encaminada a mermar el exceso de existencias, a pesar de la intensidad con que dicha política se adelantó, dejando como cifras finales la suma de 53, 581.000 sacos destruidos, solo desde 1931 hasta 193715. El total de cafetos en el mundo, durante 1936, era de 4.934, 000 árboles. De ese total, 2.570,000 pertenecían a Brasil. Con este número conseguiría una producción de 22. 533,000 sacos, mientras los demás países apenas aportaban 12.259,000 sacos al mercado mundial”16. Al recibir menos dinero por sus ventas al exterior, los exportadores y productores ligados a la suerte del mercado internacional redujeron las compras y los productores ligados al mercado interno afectados por la reducción estrecharon sus niveles de consumo. De esta forma todos los sectores de la economía brasileña se resintieron. Sin embargo, pese al ingreso monetario de Brasil en esta década, si se lo compara con otros casos como por 14 Lleras Restrepo, Carlos. “La crisis cafetera”, Artículo publicado en el Boletín Oficial de la Contraloría General de la República, Noviembre de 1937. 15 Cifras extraídas de: Lleras Restrepo, Carlos. Ibídem. 16 Taunay, Affonso de. Pequena História do Café no Brasil, (1721 – 1937), Edicao do Departamento Nacional do Café, Río de Janeiro, 1945. (pp. 492). 1 ejemplo el de los Estados Unidos, Brasil nunca presentó balances negativos en sus inversiones netas. La recuperación económica de Brasil no se debió a condicionantes externos sino a las políticas de fomento industrial seguidas conscientemente por el país, que salió con el tiempo de la dependencia que mantenía sobre el precio del café y de otras materias primas en el mercado mundial. El crecimiento de la demanda de bienes de capital, fue el reflejo de la expansión de la producción para suplir el mercado interno, y la fuerte elevación de los precios de importación de esos bienes acarreada por la depreciación cambiaria crearon condiciones propicias para la instalación en el país de una industria pesada pese a las limitaciones propias de los países periféricos o dependientes con ausencia de grandes mercados consumidores. Los negocios privados fueron también alentados a continuar sus esfuerzos, mientras el gobierno federal expandió su actividad para dirigir la economía de dos grandes maneras: por medio del manejo de estímulos, como impuestos, controles de cambio, cuotas de importación, controles crediticios y requisitos laborales; y una intervención directa caracterizada por las inversiones públicas, en áreas como los ferrocarriles, la construcción de puertos, servicios públicos e industrias básicas, como el petróleo y el hierro”17. La industrialización de Brasil, emprendida por el gobierno de Vargas desde 1930 – 1945, fue el producto de diversos factores como, la puesta en marcha de una política coherente de sustitución de importaciones, resultante del colapso de la capacidad de comprar, al mantenimiento de una demanda doméstica por medio de los programas de apoyo al café y al “(...), cambio de la inversión privada del sector exportador hacia una producción 1 industrial para surtir al mercado doméstico18”, en resumen, a la intervención directa o indirecta del Estado. Esta política estaba justificada por unos argumentos basados tanto en consideraciones económicas como de seguridad nacional. Los elementos esenciales para este paquete de medidas estaban en boga durante el ambiente de crisis general reinante en el mundo, tanto en las doctrinas corporativistas o fascistas en Europa, como en el New Deal de los Estados Unidos y en el socialismo estatal de la URSS. El nacionalismo impuesto por el militarismo dirigió la creación de una industria siderúrgica, también la metalúrgica, la de fabricación de cemento, la extractiva de minerales y la agroindustrial recibieron estímulos. Además se conformaron institutos federales para apoyar a otros cultivos diferentes al café, como el cacao, el azúcar y el alcohol. La depresión demostró que Brasil tenía pocos chances fuera de la industrialización, si buscaba transformarse así mismo en una nación moderna y una potencia mundial. Brasil gozaba de una ventaja monopólica dentro del mercado cafetero: … cuando Brasil intervenía, se protegía exitosamente contra la caída de los precios del café en épocas de cosechas mayores a lo normal, o conseguía incrementos en los precios del café cuando la oferta era menor. (...), sin embargo, las transformaciones del mercado mundial del café dieron lugar a la erosión de los beneficios de las estrategia de mercadeo brasileño, porque amparada por la costosa intervención del Brasil, Colombia entró vigorosamente en el mercado y amplió sus exportaciones”19. 17 Skidmore, Thomas E. Politics in Brasil (1930 – 1964), An experiment in democracy, Oxford University Press, New York, 1967. (pp. 43). 18 Ibídem, (pp. 47). 19 Bates, Robert. Ibídem, (pp. 67). 1 La inmersión de Colombia en el mercado internacional del café (1886 - 1934). Durante las últimas décadas del siglo XIX los cultivos de café se habían comenzado a expandir por la región central del país, desde la frontera con Venezuela en Santander propiamente, hacia la sabana de Cundinamarca. Esta región contaba hasta la Guerra de los Mil Días con el 80% de la producción nacional. Sin embargo, el deterioro en los precios en las exportaciones desde 1890 debido a la crisis internacional sufrida en aquella época, había golpeado las finanzas del Estado, que dependían de los ingresos aduaneros. Para financiar la guerra, “(...), el gobierno emitió un decreto que autorizaba a la junta emisora para emitir y poner a disposición del gobierno las cantidades que este necesitara para atender el restablecimiento del orden público”20. Con este fin se emitieron más de mil millones de pesos, lo que llevó al cambio extranjero a proporciones descomunales: desde el 412% a comienzos de la guerra al 18.900% en octubre de 1902. El descenso del tipo de cambio provocó una carestía en el abastecimiento de alimentos importados debido a los altos precios, al mismo tiempo que se encareció el transporte, por la escasez de mulas y barcos producto de la guerra. Solo el ferrocarril sobrevivió en buena medida a los rigores del conflicto. La interrupción de los transportes, el encarecimiento de los fletes, el reclutamiento forzoso de trabajadores para formar los ejércitos y el abandono de los campos para evitar el reclutamiento dejaba a Colombia a la suerte de una crisis sin precedentes, amputada en términos territoriales por la pérdida de 20 Bejarano, Jesús Antonio. El despegue cafetero, (1900 – 1928), Tercer Mundo Editores, Bogotá, 1993. (pp. 165). 1 Panamá e incapaz de encontrar una salida viable para su recuperación económica. La desaparición de fábricas e industrias en el centro del país, sobretodo en el centro tradicional del artesanado en Santander, dejó en estado de ruina la incipiente economía que se había construido a partir de la comercialización de café. La salida para muchos de los antiguos combatientes de la guerra civil estuvo destinada a la ocupación, colonización y empleo de las tierras ubicadas al noroeste de la región andina, región que había escapado de los estragos de la guerra y que se encontraba desierta de pobladores. Tanto Antioquia, como Caldas y el Valle fueron los centros de recepción de las olas de migrantes internos. De esta manera, estando el centro tradicional del país destruido, se desplazó el polo de desarrollo al eje de la cordillera occidental lo que reforzó la necesidad de construir vía férrea y el trazado de una carretera hacia el puerto de Buenaventura en el Pacífico, incomunicado históricamente del resto del país. La hegemonía conservadora había defendido la Constitución de 1886 y ganado la guerra civil, cuestión que aseguraba la permanencia de un orden jerárquico de sociedad, caracterizado tanto por la gran influencia de los comandantes vencedores como por el clero católico. A los gobiernos que siguieron tras la debacle provocada por la guerra les tocó reconstruir a un país demolido. Se adoptaron para tal fin medidas tales como la centralización fiscal, el proteccionismo económico y el impulso estatal a las actividades empresariales, y el alejamiento de algunos de los postulados económicos liberales. Se buscó una sana administración central, la estabilización del sistema monetario, el regreso al patrón oro, la restauración del crédito de Colombia en el exterior y la atracción del capital extranjero. Además del mejoramiento de los sistemas de transporte y los estímulos a la agricultura de exportación, en especial el café, el azúcar y el banano. 1 En 1923 se creó en Banco de la República cuya función central en la economía posibilitó la conversión del capital al papel moneda. El tipo de cambio se fijó en 10.000% frente al oro, se aumentaron los aranceles y se adoptaron algunas exenciones generales y particulares para la importación de maquinarias y materias primas, se establecieron subsidios, garantías sobre el rendimiento de las inversiones de capital y se permitieron las celebraciones de contratos de privilegios. El modelo proteccionista resultó ser más operativo y eficaz que el anterior modelo de economía liberal manchesteriano y creo las bases de estímulo al amparo de las cuales comenzó a consolidarse el incipiente andamiaje industrial del país. La expansión del café en las nuevas zonas de producción conllevó al nacimiento de nuevas formas de organización social y productiva, con mayores alcances sobre la estructura global del país. Entre 1906 – 1931, fueron concedidas por el estado 216.867 hectáreas de tierras en Antioquia y Caldas repartidas de la siguiente manera según Jesús Antonio Bejarano, el 12% a colonos, el 47% a individuos con propiedades de menos de 1000 hectáreas y el 41% entre los grandes propietarios. Esta repartición equilibrada era una forma nueva de repartición del ingreso y posibilitó la acumulación de capital suficiente para ser invertido tanto en la ampliación de la red ferroviaria, que pasó de 593 kilómetros en 1898, a 1143 kilómetros en 1914 y a 1571 kilómetros de cobertura en 192221, como en la construcción de una industria básica en Medellín como centro. Esta ciudad creció precipitadamente hasta romper el poder hegemónico consolidado por Bogotá tras el triunfo de la Regeneración, en 1886. Para 1916, el 70% de la industria del país se encontraba en Medellín, y estaba compuesta por 1121 establecimientos manufactureros. 1 La situación económica comenzó a cambiar gracias al ascenso en las exportaciones agrícolas y la defensa de la producción industrial. Entre 1915 – 1919 se exportaron 44.5 millones de dólares anuales, entre 1922 – 1924 se incrementaron hasta 63.9 millones anuales y para el período entre 1925 – 1929 se llegó al record de 112 millones de dólares. Este incremento paulatino pero constante fue provocado por el posicionamiento del café colombiano en el exterior y la consolidación de Colombia como el segundo productor mundial del grano. Sin embargo, pese al incremento de su producción, tan solo representaba para 1930 en 10% del mercado cafetalero global. Durante esos milagrosos años veinte Colombia había recibido otro incentivo económico que ayudó al impulso económico, los Estados Unidos habían pagado una suma de 25 millones de dólares como parte de la indemnización reconocida al país por los perjuicios ocasionados debido a la pérdida del istmo de Panamá en 1903. Además, el auge financiero vivido por los Estados Unidos le posibilitó gozar a los gobiernos colombianos de fuentes crediticias internacionales. El endeudamiento creció a pasos gigantescos en pocos años alcanzando la suma de 63.4 millones de dólares en 1926 y 203 millones en 1928. Los empréstitos habían sido otorgados en su mayor parte a las arcas departamentales y municipales que eran forzadas a cumplir sus compromisos por medio del suministro de materias primas baratas al mercado mundial. El respaldo para el pago de estas obligaciones se sostenía en la continuidad del crecimiento anual del producto interno bruto, que durante los años veinte había alcanzado un promedio anual de 7.7% y por el incremento espectacular de los ingresos estatales anuales que habían llegado a obtener 107.5 millones en 1928. Sin embargo, “(...), el gasto público se había triplicado de 5.5 21 En: Bejarano, Jesús Antonio. Ibídem, (p. 181). 2 millones a 13.7 millones anuales y la inversión pública llegó al 29.4% de la inversión total nacional, mientras en el sector de transporte la inversión fue de 74.9%”22. El ferrocarril era el elemento limitante de la economía nacional, la carencia de un sistema articulado de transportes para los productos generaba una dispersión y una fragmentación en los mercados. Hasta entonces la función de los ferrocarriles había sido la de integrar las áreas cafetaleras con los puertos de embarque para el exterior, se había configurado un sistema centrífugo en la infraestructura vial, es decir, economías de enclave. Para 1922 los 1481 km. de red ferroviaria constaban de 992 km. localizados principalmente en las regiones cafetaleras del occidente del país y las vías correspondientes a la articulación de éstas con el puerto de Buenaventura. Los restantes estaban orientados a la comunicación del centro con los puertos de embarque en el Caribe o conectadas a regiones no vinculadas con el comercio internacional. Durante los años veinte se trató de orientar la construcción de ferrocarriles, en orden de crear y facilitar el comercio interno y el mercado nacional, en pocas palabras, promover una red de tendencias centrípetas. En 1926 – 1930 se construyeron 942 km. nuevos en carreteras y caminos vecinales. La construcción del sistema vial posibilitó el incremento de la exportación de sacos de café hasta sobrepasar la cifra de 2 millones de sacos (62.5 kg.) en 1926. La mayor parte de estas exportaciones fueron enviadas al mercado norteamericano, por 132.140,664 kg., mientras el segundo mercado, Alemania solo captó 1.649,380 kg. ese mismo año23. 22 En: Bejarano, Jesús Antonio. Ibídem, (p. 195). En: Monsalve, Diego. Colombia Cafetera, información general de la república y estadísticas de la industria del café, Artes Gráficas, Barcelona, 1928. 23 2 El destino manifiesto de Colombia era avanzar hacia la colonización de tierras despobladas adaptables al café, sembrar los árboles e incrementar la producción de 600 mil sacos a 1 millón, a 5 millones, a 10 millones, para dominar el mercado de cafés suaves y aún reemplazar a Brasil”24. En respuesta al auge en los precios del grano, provocado por la intervención de Brasil en los mercados cafeteros, la producción de café aumentó. Como ya se dijo anteriormente, el centro geográfico de la industria se había trasladado de la región oriental a la región occidental, y por lo tanto, la importancia de los pequeños propietarios crecía en la medida que estos se agrupaban en torno a agremiaciones de productores. La más importante de aquellas, la Federación Nacional de Cafeteros, incorporó no solo a los campesinos, pequeños y medianos productores sino a elementos claves, pertenecientes de la influyente clase política conservadora de las regiones cafetaleras, quienes mantenían un control importante sobre el mercado del café. La FNC, creada por políticos locales y regionales fue dotada del poder público para castigar el comportamiento oportunista. Se le dio el poder para obligar a sus miembros a actuar coordinados de tal forma que aumentara la productividad de la industria en su conjunto, convirtiéndose en una institución económica de primer orden. La FNC optó desde sus principios por una estrategia competitiva del café colombiano en el extranjero, siendo esto el resultado de una lucha política interna contra los gobiernos de la república liberal entre 1930 – 1938. Pese a los obstáculos generados por las nuevas fuerzas democráticas, las presiones sociales, producto de las reivindicaciones salariales, las garantías laborales, y las reformas en el manejo económico impuestas por el nuevo rumbo 24 Carlyle Beyer, Robert. The Colombian Coffee Industry. Origins and mayor trends, Tesis de Doctorado, 2 económico del liberalismo reformista, los caficultores lograron incidir de forma influyente sobre la política económica nacional. Mediante la ley 21 de 1927, el gobierno colombiano impuso un impuesto a las exportaciones de café, cuyo recaudo quedó en manos de la FNC, los cafeticultores obtuvieron con ello el poder del Estado, en la medida en que las autoridades aduaneras y las oficinas de impuestos ahora se desempeñaban como agentes de la organización. Al interior de las regiones cafeteras, se buscó proveer la infraestructura básica: almacenes de insumos, facilidades de crédito e información sobre los precios de los mercados externos y asistencia técnica en las nuevas tendencias de producción y comercialización del café. Además se enfatizó en una comercialización del café dependiente de su lugar de origen. La selección y clasificación de diversos tipos de café, generó un incremento en los precios y las marcas definieron las calidades de los granos y por lo tanto su estatus dentro del mercado internacional. El compromiso de la Federación con una campaña internacional que fomentase el consumo del café colombiano, conllevó a que se firmaran contratos de exclusividad con agencias de publicidad encargadas de promocionar las ventas en tiendas especializadas, cadenas de almacenes, restaurantes y para el consumo del hogar o en los lugares de trabajo. Algunos de los lugares en donde se golpeó con esta nueva estrategia comercial fueron las ciudades europeas de Madrid, Barcelona, París, Bruselas, Berlín, Praga, Roma y Nápoles. También se establecieron oficinas permanentes en Nueva York y Londres para analizar las fluctuaciones de los precios y la demanda internacional del café. Además, la Federación introdujo en el Brasil, un agente del gobierno, a quien nombró como cónsul en Sao Paulo, Universidad de Minnesota, 1974. (pp. 181). 2 cuyo principal encargo era enviar informes sobre la industria cafetera de Brasil a la sede de la Federación en Bogotá. Los primeros años de la gran depresión estuvieron dominados en Colombia por el colapso del mercado del café y por la brusca interrupción de los flujos de capital. Los precios del café empezaron a disminuir desde los primeros meses de 1929, en octubre de ese año colapsó la bolsa de valores de Nueva York y la política de ajuste que adoptó el gobierno brasileño dejaron al estado de Sao Paulo sin recursos financieros para mantener la intervención en defensa de los precios. La crisis de 1929 arrastró también los precios de los demás productos agroindustriales, que constituían el grueso de las importaciones del país. El colapso del café y de otras materias primas fue aún más pronunciado, generando una base del 23% en los términos de intercambio de Colombia, entre 1925 – 1929 y 1930 – 193425 ”. El impacto del deterioro de los términos de intercambio prevaleció, provocando una merma del 8% en el poder de compra de las exportaciones entre los años de bonanza y la primera mitad de la década del treinta. Ésta caída, fue moderada e insuficiente para desencadenar una crisis de grandes proporciones. En 1933, el precio del café Manizales en Nueva York se cotizaba en 10.5 dólares por libra, apenas una fracción de su nivel de bonanza de 28.5 dólares por libra en 1926. Desde 1930 el presidente Olaya Herrera, emprendió una campaña entre los círculos financieros norteamericanos para conseguir préstamos de corto plazo apoyándose, en su política de apertura a las inversiones petroleras. 2 De esta manera logró obtener crédito por solo 17 millones durante su primer año de mandato, una cantidad insuficiente, ya que el pago de la deuda externa sumaba la cifra de 19 millones de dólares anuales”26. Sin embargo, Colombia, como la mayor parte de los países latinoamericanos se vio forzada a declarar la moratoria sobre el servicio de la deuda externa a comienzos de los años treinta, a pesar de los esfuerzos del gobierno por mantenerse dentro de las reglas de juego de la banca internacional. Los primeros pasos hacia la moratoria se dieron a raíz de la implementación del control de cambios en septiembre de 1931. En octubre de 1931 se decidió que los pagos a la deuda externa de los departamentos, municipios, y bancos deberían ser autorizados por la Oficina de Control y Cambios. En diciembre de ese mismo año, el gobierno llegó a un acuerdo con la mayoría de los tenedores de la deuda externa colombiana, por medio del cual se suspendían temporalmente las amortizaciones de todas las deudas y se permitía pagar los intereses de aquellos que no estaban garantizadas por la nación con vales a tres años e intereses del 6%. El gobierno nacional comunicó el 23 de mayo de 1932 a la prensa norteamericana que solo seguiría pagando los intereses de la deuda nacional y la del Banco Agrícola Hipotecario, que estaba garantizado por la nación. Desde 1932 se dejó de pagar suma alguna por el 43% de la deuda externa de largo plazo que tenía el país, lo cual representaba un ahorro de 9 millones de dólares al año. A pesar de la oposición, los intereses de la deuda nacional se cancelaron oportunamente hasta principios de 1933, cuando la guerra contra el Perú llevó al gobierno nacional a suspenderlos en su totalidad. 25 Ocampo, José Antonio. “La crisis mundial y el cambio estructural (1929 – 1945), En: Historia económica de Colombia Tomo I, Cap. VI, Bogotá 1986. (pp. 207). 2 La moratoria sobre la deuda externa posibilitó una recuperación de las importaciones reales y facilitó el proceso de reactivación de la economía. En el mercado cafetero, el país no solo logró consolidar su posición en el mercado norteamericano, sino que logró grandes avances en el alemán, gracias a un acuerdo de compensación comercial firmado a mediados de la década. Con el ascenso de la República Liberal en 1930, comenzaron los enfrentamientos entre los industriales cafeteros y los gobiernos de Olaya Herrera y Alfonso López Pumarejo. Los conflictos fueron enmarcados por tres elementos: a) La valorización de la moneda, b) Los gravámenes fiscales impuestos por los gobiernos y c) La política comercial del grano a nivel internacional. La sobre valuación de la moneda colombiana, servía como estrategia para cubrir las deudas del país con los bancos extranjeros. Como la deuda debía ser pagada en divisas, “(...), una depreciación de la moneda aumentaría los costos fiscales del reembolso a los acreedores27”. La Federación en contraposición a las tendencias marcadas por el Estado lanzó una vigorosa campaña para lograr la depreciación de la moneda nacional. La presión de la FNC surtió el efecto esperado, ya que en 1932 el peso se depreció un 10% y en marzo de 1933 se devaluó nuevamente, hasta obligarlo a la adopción de la libre convertibilidad de la moneda en 1934. Esto permitió la estabilización de los ingresos provenientes de las exportaciones. Los sectores agroindustriales pedían además la reducción de los aranceles y las tarifas de transporte para las importaciones de alimentos. Las vigorosas presiones de los exportadores de café defendieron sus fortunas, a expensas 26 Ibídem, (pp. 212). Ocampo, José Antonio. Crisis mundial, protección e industrialización, Alianza Editorial, Bogotá 1984. (pp.140).	5,911
dumas-04317626-Psychomot_2023_Engleraud.txt_1	French-Science-Pile	Open Science	Various open science	2,023	"Un corps sans limites": l'apport de la psychomotricité dans la prise en charge des troubles du comportement chez l'enfant à la suite d'un défaut de construction des enveloppes psychocorporelles. Psychologie. 2023. ⟨dumas-04317626⟩	None	French	Spoken	8,145	12,682	”Un corps sans limites”: l’apport de la psychomotricité dans la prise en charge des troubles du comportement chez l’enfant à la suite d’un défaut de construction des enveloppes psychocorporelles -disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. UNIVERSITÉ de BORDEAUX Collège Sciences de la Santé- Institut Universitaire de Sciences de la Réadaptation Institut de Formation en Psychomotricité Mémoire en vue de l’obtention du Diplôme d’État de Psychomotricien « Un Corps sans Limites » : L’apport de la psychomotricité dans la prise en charge des troubles du comportement chez l’enfant . Engleraud Elsa e Maître de Mémoire : ROUYER Amandine Remerciements Je souhaite remercier toutes les personnes qui m’ont aidée et soutenue pour l’écriture de ce mémoire : Amandine Rouyer pour m’avoir accompagnée dans la réalisation de ce mémoire. Je la remercie pour sa disponibilité, ses conseils qui ont contribué à alimenter ma flexion. Sophie Cardey qui m’a supervisée au cours de mon stage en troisième année au sein de l’ITEP. Marie Daguerre qui m’a accompagnée lors de mon stage en EHPAD. Merci pour votre patience, votre précieuse aide et votre implication. Tous mes maîtres de stages, qui m’ont accueillie sur leur lieu de travail et qui ont contribué à faire de moi la psychomotricienne que je deviendrai. Merci à l’Institut de Formation en Psychomotricité de Bordeaux pour cette formation riche. Je souhaite également remercier ma famille et mes amis pour leur soutien et leur compréhension tout le long de ma formation. 1 Avant propos « Le dé faut ou les défaillances de cette première enveloppe, de ce premier container psycho corporel entraîne souvent une organisation psychopathologique ou des défenses massives de type psychotique. » (COURBERAND Daniel, 2015) 2 Sommaire ● Remerciements................................................................................................1 ● Avant propos...................................................................................................2 ● Introduction....................................................................................................4 ● Partie 1 : Le développement de l’enfant I - Le développement cognitif...........................................................................6 II - Le tonus.....................................................................................................11 III- Le concept de transitionnalité...................................................................16 ● Partie 2 : Structuration des enveloppes psychocorporelles I - L’enveloppe psychique et fonction contenante............................................17 II- Le corps et ses représentations..................................................................27 ● Partie 3 : Psychopathologie de l’enveloppe I- Défaut de contenance et troubles du comportement...................................38 II- Définition des troubles du comportement.................................................41 ● Partie 4: Rôle de la psychomotricité I- Présentation de l’institut thérapeutique éducatif et pédagogique................50 II- Cas cliniques............................................................................................53 ● Conclusion.....................................................................................................83 ● Bibliographie.................................................................................................85 ● Table des matières......................................................................... ................ 90 ● Annexes........................................................................ ................................92 3 Introduction Que sont les enveloppes corporelles? Les enveloppes psychiques? Comment structurent-elles l’individu? Quel est leur lien avec les troubles du comportement? Comment pouvons-nous, en tant que psychomotricien, remédier à ce défaut de structuration? Je tenterai de répondre à ces différentes interrogations tout au long de cet écrit avec un point de vue psychomoteur. L’idée de ce sujet m’est venue progressivement ; tout d’abord mon envie de thème de mémoire portait sur le toucher puis mon stage de troisième année a débuté. Mon idée s’est alors orientée sur le concept de contenance et de limites corporelles. Le toucher est fortement lié à ces concepts et sera donc évoqué tout au long de ce mémoire. J’effectue mon stage de Troisième année au sein d’un Institut thérapeutique éducatif et pédagogique (ITEP), la population rencontrée en psychomotricité est majoritairement des enfants, entre 5 et 8 ans, présentant des troubles du comportement et de la relation. J’ai constaté rapidement que nombre de ces enfants présentaient un point commun : un défaut de construction des limites corporelles, un défaut de contenance. Cette structuration défaillante entraîne alors certains comportements atypiques, une relation particulière, une instabilité psychomotrice. Il semblerait que le sentiment de ne pas avoir été contenu physiquement et psychiquement peut alors induire des troubles du comportement. Par l’observation et le suivi de certains patients, j’ai donc pu observer l’apport de la psychomotricité dans ce type de troubles. 4 De cette réflexion est née la problématique suivante : Comment la chez médiation psychomotrice les enfants présentant des troubles du comportement en ITEP participe-t-elle à la restructuration de leurs enveloppes psychocorporelles? Dans un premier temps, j’exposerai le développement de l’enfant avec l’importance de l’environnement et des expériences sensorielles, tout en incluant le rôle du tonus dans ce développement et la notion d’espace transitionnel. Dans un second temps, je développerai comment la conscience corporelle s’édifie avec notamment les notions de contenance, de « holding » de Winnicott, le « Moi Peau » d’Anzieu ou même la théorie de l’attachement. J’aborderai également les notions de schéma corporel et d’image du corps, tout en resituant l’importance du corps en psychomotricité. Dans un troisième temps, je définirai certains troubles du comportement et j'établirai leur lien avec la psychopathologie de l’enveloppe. Dans un quatrième et dernier temps, j’expliquerai brièvement le principe et le rôle d’un ITEP pour ensuite détailler deux cas cliniques rencontrés. 5 Partie 1 : Le développement de l’enfant L’étude du développement de l’enfant est nécessaire afin de comprendre les facteurs qui contribuent à une structuration psychique dite “normale”. I- Le développement cognitif Au vu de la population rencontrée et de l’âge des patients présentés en étude de cas, je détaillerai davantage les stades se situant entre 0 et 8 ans. Les autres stades seront évoqués de façon plus succincte. a- Selon J.Piaget J. Piaget, psychologue, a étudié les différentes phases du développement de l’enfant afin de comprendre l’intelligence humaine. Il décompose ce développement en 4 stades dépendant de l’environnement dans lequel l’enfant est inséré. Tous ces stades se déroulent dans le même ordre pour chaque enfant (mais pas forcément aux mêmes âges). Il étudie donc l’intelligence de l’enfant qui va passer d’une intelligence sensori motrice à une intelligence abstraite. Pour lui, c’est surtout l’action sur son environnement qui va permettre de construire les structures de l’intelligence. 6 1- Le stade sensori-moteur, de la naissance à environ 2 ans Ce stade est caractérisé par le développement des capacités sensorielles et motrices du bébé. En effet, il va interpréter le monde environnant selon ses sens et ses actions avec la constitution des schèmes sensori-moteurs. Au début, le bébé va travailler ses réflexes. Puis petit à petit, l’enfant a adapté progressivement ses réflexes, on parle de réactions circulaires primaires. Il pouvoir répéter des comportements dont les effets lui procurent du plaisir. Il joue et expérimente différentes parties de son corps. Ainsi, au fur et à mesure il va donner du sens à de l’expérience. A l'issue de ce stade, l’enfant peut reconnaître son corps, comprendre la relation de cause à effet et la notion de permanence de l’objet mais aussi avoir une représentation mentale des objets et des événements. 2- Stade préopératoire, de 2 à 6 ans Cette période est marquée par le développement de la représentation symbolique. Elle va se manifester à travers le langage, l’imitation différée, le dessin et le jeu. A travers ce jeu symbolique, il se crée soit un monde fictif où il se laisse déborder par son imagination soit un monde proche du sien. Ce jeu aide parfois à résoudre des conflits, d’où l’importance et l’observation du jeu libre en psychomotricité. Il tente de comprendre les causes des événements, de percevoir les émotions, de mettre en place des stratégies pour atteindre un but. C’est également l’âge d’une plus grande intériorisation de l’action. 7 3- Le stade des opérations concrètes, de 7 à 12 ans L’enfant à cette période a des capacités cognitives importantes qui se développent de plus en plus. Il peut envisager d’autres points de vue que le sien. Il a la capacité de faire des opérations mentales de façon logique pour résoudre des problèmes. Il recherche des explications logiques au lieu de faire appel à ces intuitions. 4- Le stade des opérations formelles, plus de 12 ans Les adolescents sont dotés de nombreuses habiletés même s' il existe une énorme variabilité du fonctionnement cognitif. Ils ne sont plus obligés de passer par le concret et peuvent réfléchir sur des notions abstraites et des notions en soi (le bien, le mal, l’infini, la mort...) b- Selon L. Vygotsky Connu pour ses recherches en psychologie du développement et notamment sa théorie historico-culturelle du psychisme, son objectif est l’étude du développement des fonctions psychiques supérieures. Il se base sur trois points: ★ Approche historico-culturelle, cela dépend de la société où on évolue ★ Ancrage social du développement, se fait dans les relations sociales ★ Instruments psychologiques, c’est le langage, l’écriture... (qui s’acquièrent à travers les intéractions). Pour lui, il y a une relation entre le groupe social et l’environnement de l’enfant avec le développement de son intelligence. On apprend et on se développe dans l’échange avec les autres. 8 c- Selon H. Wallon Henri Wallon, psychologue et neuropsychiatre, cherche à comprendre à travers les dimensions sociales, affectives et cognitives le développement de la personnalité. Il définit 6 stades correspondant à des étapes importantes du développement aboutissant à l’intégration de l’affectivité et de l’intelligence. Le développement résulte de l'interaction avec l’environnement. 1- Le stade impulsif, de 0 à 3/6 mois Ce stade est prédominé par le facteur affectif et caractérisé par un désordre gestuel, une fusion émotionnelle. Le bébé dépend totalement de ses parents et du milieu extérieur. 2- Le stade émotionnel, de 3 mois à 12 mois Les émotions se différencient petit à petit à l’aide des réponses adaptées et répétées de l’entourage. Ceci lui de préciser sa gestuelle et d ‘adapter ses réactions. 3- Le stade sensorimoteur et projectif, de 1 à 3 ans L’enfant va développer le langage et la marche. Il explore son environnement, le corps est un outil indispensable pour réaliser ses différentes expériences. Vers la fin de ce stade, on parle de “stade projectif” car l’enfant projette sa pensée grâce aux gestes. 9 4- Le stade du personnalisme, de 2 à 6 ans Ce stade marque la découverte de l’opposition et de la négociation et teste le cadre et les limites. Il adapte son comportement en fonction des personnes avec qui il interagit. C’est aussi le début de la différenciation entre le moi et autrui. 5- Stade catégoriel, de la personnalité polyvalente, de 6 à 11 ans Le développement de l’enfant sera davantage guidé par l’intelligence. Ce stade est aussi marqué par la capacité de l’enfant à se représenter les choses de manière abstraite. 6- Stade de l’adolescence, à partir de 11 ans Les conduites affectives prédominent. On se concentre sur soi et découvre le nouveau corps lié à la puberté. Cette période permet d’achever la construction de la personnalité. Wallon considère donc le corps comme un support pour les intéractions entre le monde social et l’enfant. Par exemple, les émotions de bébé sont identifiées par l’environnement à partir de son tonus et de son activité motrice (hypertonie, hypotonie). Ces intéractions précoces entre la mère et l’enfant permettent aussi à l’enfant de prendre conscience de lui, grâce à l’autre. 10 II- Le Tonus Le tonus est l’une des pierres angulaires de la psychomotricité. Il me paraît donc incontournable d’en évoquer son intérêt. C’est un “état de tension active, permanente, involontaire et variable” (Scialom et al., 2011) Le tonus est sous la dépendance de deux types de motoneurones et de boucle gamma. Il existe différents types de tonus : tonus de fond, tonus postural et tonus d’action. Il constitue le fondement même de la posture, du mouvement et des dynamiques d’interaction avec l’environnement. En effet, le tonus est d’une importance fondamentale dans l'organisation émotionnelle de l’enfant et dans les étapes de son développement. Il est propre à chaque individu et reflète la personnalité, la manière d’être et d’agir de chacun face à l’environnement. Il intervient également dans la relation avec autrui. Le tonus est la toile de fond des activités motrices et posturales. Au cours du développement et à l’âge adulte, le tonus est un aspect fondamental du bien être chez un individu. a- La maturation du contrôle tonique J. Ajuriaguerra définit 3 stades du développement psychomoteur de l’enfant. ● de la naissance à 3 ans Le tonus est réparti de manière bipolaire: - hypertonie (contraction exagérée) lorsqu' il est dans état de besoin - hypotonie (tension à peine perceptible) lorsqu’on subvient à ses besoins 11 Aussi, le contrôle tonique est dépendant des personnes qui vont s’occuper du nourrisson. C’est la présence, la satisfaction de ses besoins et la consolation par son entourage qui abaisse ses tensions. L’édification tonique est également dépendante de la maturation neurologique avec une équilibration globale du tonus des muscles. Effectivement, au début, le nouveau né présente une répartition tonique déséquilibrée avec une hypotonie axiale et une hypertonie des muscles fléchisseurs. Le développement du contrôle du tonus va dans le sens de la myélinisation et suit également les lois céphalo-caudales et proximo-distales. Donc “l’enfant sort de la bipolarité émotionnelle primaire et globalise ses tensions, ses sentiments et ses représentations opposées dans une même enveloppe psychique. ” (Scialom et al., 2011) En effet, cette maturation du contrôle tonique est sous dépendance de facteurs psychologiques et physiologiques. ● de 3 à 6 ans La structuration perceptive joue un rôle prépondérant lors de ce stade, avec pour objectif d’arriver à un contrôle de sa tonicité. On constate aussi une inhibition des réflexes archaïques. Il va apprendre à utiliser les différentes stimulations visuelles ou sonores et adapter ses réponses motrices et toniques. Comme par exemple, marcher sans faire de bruit ou adapter sa force lors de lancer de ballons. Enfin vers l’âge de 4-5 ans, l’enfant est en capacité d’expérimenter une détente globale en interrompant ses contractions musculaires. 12 Il sera en mesure de contrôler plus consciemment ses états toniques. ● de 6 à 12 ans Le processus de conscience corporelle débuté dès la naissance mais c’est vers l’âge de 6 ans que la représentation corporelle s’actualise. Grâce à ses compétences à relâcher ses différents membres, l’enfant va se construire une certaine topographie du corps (le schéma corporel). Tout ceci contribuant à l'organisation mentale du temps, de l’espace et du geste. b- Rôle du tonus Différentes fonctions peuvent être attribuées au tonus, je vais donc en citer certaines. Tout d’abord, le tonus joue un rôle dans la conscience de soi et la perception corporelle. La contenance tonique procure un sentiment d’unité corporelle. De par son lien avec la proprioception, il instaure aussi la conscience d’être en mouvement, d’être en relation. Ainsi, la prise de conscience de ces états toniques, de son enveloppe tonique participe à cette conscience de soi. Ce fond tonique relie toutes les différentes parties de notre corps. Il permet la cohésion de l’ensemble du corps. De plus, le tonus est le support de notre système émotionnel. “ Il est la chair de l’émotion” (Scialom et al., 2011) La régulation émotionnelle ne peut être dissociée de notre régulation tonique. Nos empreintes affectives sont marquées au sein de notre tonus. 13 Lors d’expériences émotives, de rencontres à autrui, l’observation du tonus est importante. Par exemple, en situation de peur, de stress une hypertonie est observable. En tant que psychomotriciens, nous sommes très sensibles à cette lecture du corps, du tonus pour comprendre l’état émotionnel dans lequel se présente notre patient. En raison de son lien avec la sphère émotionnelle, il en découle une autre fonction: celle de communication. Pour le nouveau né, c’est une voie privilégiée et unique de communication avec sa mère au sein du dialogue tonico-émotionnel. De l’enfance à la mort, cette fonction de communication persiste. Elle peut autant être consciente que inconsciente. A travers notre corps, notre visage et nos postures, nous véhiculons certains messages ( observation des variations toniques). “La tonicité est un baromètre interne” (Scialom et al., 2011) Les variations entre états hypertoniques et hypotoniques sont révélatrices. L’hypertonie est associée à des sensations d’angoisses, de doutes, de conflits ou alors à de l’excitation et du désir. L’hypotonie est liée à des états de dépression et de détresse mais aussi à de la détente et de la tendresse. c- Le dialogue tonico-émotionnel Le dialogue tonico-émotionnel serait le reflet des états émotionnels des deux partenaires avec la possibilité d’une transmission de l’un à l’autre, en particulier chez le bébé. Lorsque le bébé est dans les bras de sa mère, il est particulièrement sensible aux variations toniques et rythmiques des bras qui portent et l’entourent. 14 C’est là que plusieurs paramètres doivent être pris en compte : le tonus des bras, le rythme du portage, le visage, la prosodie... Grâce au dialogue tonique, le bébé accumule et intègre des informations riches, variées et adaptées. Ceci lui permettra de reconnaître ses propres éprouvés et de les différencier de ceux des autres. Le sujet vit son corps et interagit dans la relation à l’autre à travers ce tonus. Ce jeu du dialogue tonique va se vivre et s’observer à plusieurs niveaux et on retrouvera ces notions que Winnicott a développées c’est-à-dire dans les qualités de soins données à l’enfant par le holding, le handling et l’object presenting. Ce jeu est également retrouvé auprès de nos patients et représente un outil thérapeutique important. Il permet aussi l’intégration des sensations, par la confrontation aux différents états toniques des personnes rencontrées. Enfin, le sens donné aux émotions et tensions ressenties par le bébé permet d’accéder à la symbolisation. Parfois, la communication tonique se fait mal, c’est un mauvais dialogue. Ce sont des réponses non adaptées ou une absence de réponse, favorisant ainsi une hausse tonique. On comprend bien ici, l’enjeu de cette maturation tonique à travers le dialogue tonico-émotionnel notamment. C’est un enjeu du point du vu affectif et développemental. La capacité d’auto-régulation est également dépendante des réponses durant cette prime enfance. 15 apportées III- Concept de transitionnalité Nous attribuons ce concept à D. Winnicott avec les notions “d’objet transitionnel” et “d'aire transitionnelle”. Ces concepts sont considérés comme clés dans le développement de l’enfant notamment dans le développement émotionnel et psychologique. L’objet transitionnel est une notion ayant teint le grand public. En effet, l’intérêt du doudou est connu par tous. Celui- ci permet de mieux supporter l’absence de la mère. La notion qui m’intéresse ici est celle d’aire, d’espace transitionnel. Elle correspond à un espace qui n’est ni moi, ni non moi, ni dehors, ni dedans. C’est une aire où se réalisent des expériences et qui participe au développement psychique. Elle permet de laisser place au jeu, à la créativité, à l’agressivité de l’enfant. Elle fait le lien entre la réalité interne et la réalité du monde extérieur. C’est un espace sécure, rassurant et partagé. L’espace créé en psychomotricité s’apparente à l’espace transitionnel de Winnicott. L’espace thérapeutique représente un lieu où l’enfant est censé se sentir en sécurité pour expérimenter, décharger... Il fait le lien entre les expériences vécues et les représentations, la transition entre monde intérieur et extérieur, autrement dit la symbolisation. La notion d’aire transitionnelle se confond donc avec la notion du cadre thérapeutique. Ce cadre, de part ses limites et la contenance, joue un rôle d’enveloppe et représente un lieu d’expérimentation. Le thérapeuthe participe à ces bonnes conditions. Il a pour rôle d’instaurer et de maintenir ce cadre comme les soins parentaux favorisent la structuration des enveloppes psychocorporelles. 16 Partie 2 : Structuration des enveloppes psychocorporelles L’enveloppe psychocorporelle est l’union de l’enveloppe corporelle et de l’enveloppe psychique. En unissant le corps et le psychisme, cela fait d’elle une notion psychomotrice. I- L’enveloppe psychique et fonction contenante a- Premiers modèles post-kleinien sur la notion d’enveloppe Le concept d’enveloppe psychique se retrouve au sein de plusieurs disciplines telles que la psychanalyse, l’étude du développement psychique de l’enfant, la neuropsychologie et les sciences cognitiv es. Le domaine de la psychanalyse est riche en littérature concernant ce concept. Il permet de décrire certaines organisations psychopathologiques, de mieux comprendre la genèse et certains modes de fonctionnement. Ce concept a également enrichi la fonction du cadre thérapeutique sur lequel je reviendrai. Qu’est-ce qu’une enveloppe? C’est ce qui entoure complètement un espace, contient tout ce qui se trouve dans celui-ci, par exemple, l’enveloppe cutanée d’un être vivant. Cette enveloppe définit une limite entre un dedans et un dehors. L’enveloppe psychique n’est pas un objet mais a un rôle de fonction : la fonction contenante. Cette fonction contenante est déléguée à la personne qui va s’occuper de l’enfant. Elle existe dans une rencontre. Dès 1962, Bion construit le modèle du « contenant-contenu » ; l’expérience du bébé nécessite la présence d’un contenant pour accueillir, transformer et détoxiquer les différentes informations de l’environnement. 17 Cette détoxification réalisée le plus souvent par la mère porte le nom de « fonction alpha ». L’objet contenant (par exemple la mère) reçoit les informations brutes « les éléments bêta » et les transforme en « éléments alpha » pour l’enfant. Ces éléments transformés seront disponibles pour la pensée du bébé. Le bébé n’étant pas en capacité d’assurer lui-même la transformation nécessaire de ses éprouvés corporels pour les assimiler psychiquement. Cette « fonction alpha » est d’abord dévolue par la mère, peu à peu l’enfant intériorise et devient capable de traiter lui-même ses propres expériences pour les transformer en pensées. Donc au début la mère va « prêter » son appareil « à penser les pensées » jusqu’ à ce que l’enfant se construise le sien. Il y a quelque chose de l'ordre des impressions que le bébé va ressentir (faim, sommeil...) qui précède à la sensation, qui n’ont pas de sens pour lui au départ. Elles vont être contenues dans ce giron maternel. Cela sera sous forme de portage, de dialogue tonico-émotionnel... C’est à partir de cette incapacité à lier ses impressions, que le bébé va créer une demande, vécue par la mère comme un appel. Elle va contribuer par ses actions à lier ses ressentis et faire que le bébé puisse se développer. Or pour en arriver à ce stade, il faut que l’objet contenant ait été en capacité de se rendre disponible et d’apporter des réponses adaptées afin de démarrer cette transformation. 18 Esther Bick, en 1967 introduit le concept de « peau psychique ». « La thèse est que, sous leur forme la plus primitive, les parties de la personnalité sont ressenties comme n’ayant aucune force liante entre elles et doivent de ce fait être tenues passivement ensemble grâce à la peau faisant office de limite. Mais cette fonction interne de contenir les parties du Self dépend de l’origine de l’introjection d’un objet externe, reconnu apte à remplir cette fonction » Esther Bick en 1968, (Houzel, 2010). L’objet externe et contenant jouant cette fonction est le mamelon dans la bouche, accompagné par le portage, les paroles, les odeurs familières... Les soins parentaux jouent un rôle indispensable dans la contenance et la construction de limites. Le nourrissage représente une expérience de rassemblement à l’intérieur d’une peau. Esther Bick nous dit que faute de cette expérience d’une relation à objet contenant extérieur alors l’enfant s’engage dans la constitution d’une seconde peau psychique. Elle s’apparenterait à une sorte de coquille protectrice formée par un surinvestissement de la musculature protégeant le bébé contre des angoisses. Donc Esther Bick montre comment cet objet contenant est éprouvé comme une peau. Les perturbations de cette première peau conduisent au développement d’une seconde peau, dans laquelle l’indépendance envers l’objet est remplacée par une pseudo-indépendance, en créant un substitut à cette fonction de contenant-peau. Enfin, Geneviève Haag démontre que le contact bouche-sein combiné au contact œil à œil entre le bébé et sa mère crée des boucles de retour c’est-à-dire comme si en « partant de l’enfant, un lien atteignait la mère, notamment son regard, pour faire retour vers l’enfant à partir d’un point d’inflexion qui correspondrait au sentiment d’avoir pénétré le psychisme de la mère » (Houzel, 2010) Ce lien entre qualité de contact du regard et le contact mamelon-bouche ainsi que le portage donne l’expérience au bébé d’un appui dorsal, d’une unification de son enveloppe psychique. 19 La conscience de la présence d’un arrière-plan serait expliquée par cette qualité de portage, le regard pendant l’allaitement effectuant des boucles de retour. Cette conscience est donc dépendante de la présence d’un portage satisfaisant et agréable. La représentation d’un appui dorsal est également constitutive d’un narcissisme sain. « Le bébé accède au sentiment de rassemblement de l’espace avant/ arrière lorsque la maman le porte dans ses bras » (Houzel, 2010) Cette notion de portage met en avant une prise de conscience pour le bébé d’un corps unifié et fermé. Par exemple, quand le toucher au niveau du dos est insupportable pour le patient, on peut supposer que cet arrière-plan de sécurité n’a pas été intériorisé, intégré. La contenance, c’est ce qui va donner une unité à la fois au corps mais aussi à toutes les composantes qui vont faire de nous ce que nous sommes. b- Anzieu et le moi peau Le « Moi- Peau » est le concept créé par le psychanalyste D.Anzieu, ce concept est utilisé pour parler d'enveloppe psychique. Il reprend en partie l’ensemble des modèles existants. Tout d’abord nous allons commencer par définir ce qu’est la peau selon D.Anzieu. Il choisit de définir l’univers tactile et cutané selon différentes approches : linguistique, physiologique, évolutionniste, histologique, psycho-physiologique, interactionniste et psychanalytique. Je choisirai ici d’évoquer l’approche physiologique. 20 La peau, de par sa structure et ses fonctions est plus qu’un organe, c’est un ensemble d’organes différents. Elle regroupe différents organes de sens (toucher, pression, douleur, chaleur...) et est en lien avec les différents sens externes (ouïe, vue, odorat, goût). La peau remplit également des rôles annexes, elle est le support du tonus, elle stimule la respiration, la circulation, la digestion... Elle maintient le corps autour du squelette, protège contre les agressions extérieures et transmet des informations utiles. La peau est un organe vital qui joue un rôle d’enveloppe. Anzieu a étudié les contenants psychiques à travers notre principale enveloppe physique : la peau. Par le nom de Moi-peau, il désigne « une figuration dont le Moi de l’enfant se sert au cours des phases précoces de son développement pour se représenter lui-même comme Moi contenant les contenus psychiques, à partir de son expérience de la surface du corps. »(Anzieu, 1995) Le Moi-peau trouve son étayage sur les différentes fonctions de la peau. Il définit alors 8 fonctions du Moi-peau : 1- La Maintenance : la peau remplit la fonction de soutènement du squelette et des muscles, le Moi-peau lui remplit une fonction de maintenance du psychisme. Cette fonction s’assimile à la notion de « holding » de Winnicott, elle retient le bon et le plein des soins maternels qui permet ainsi l’émergence de la pensée. 2- La Contenance : cette fonction est exercée principalement par le « handling » maternel. 3- Pare-excitation : la mère assure une pare-excitation pour son enfant. Elle le protège des excitations du monde extérieur jusqu’à ce qu’il trouve un 21 étayage corporel suffisant pour satisfaire cette fonction. C’est une fonction de protection des agressions de l’autre. 4- Individuation : cette fonction permet l’émergence du soi et de l’unicité de l’individu. 5- Intersensorialité : le Moi-peau est une surface reliant les différentes sensations grâce à notre enveloppe tactile. 6- Soutien de l’excitation sexuelle : due à la présence de zones érogènes 7- Recharge libidinale 8- Inscription des traces sensorielles : à travers les sensations tactiles, vont s’inscrire sur la peau les différentes expériences du bébé. Ce concept de Moi-peau maintient le lien entre psyché et soma. Donc pour Didier Anzieu, la peau ne se restreint pas à un rôle d’enveloppe physiologique. Elle a une fonction psychologique qui permet de contenir, de délimiter, de mettre en contact, d’inscrire... Cette fonction est tributaire de la qualité des soins maternels développés par Winnicott. c- La question du « holding » selon Winnicott Winnicott, pédiatre et psychanalyste britannique met l’accent sur nécessité d’un environnement bon pour le développement de l’enfant. A travers ses différents ouvrages “La relation parent-nourrisson” et "La mère suffisamment bonne” et articles, il évoque l’importance des soins parentaux. Il introduit les notions de « holding » et de « handling ». Le « Holding » est le tout premier stade de la relation « parents-nourrisson ». Il fait l’hypothèse que le « moi » de l’enfant n’est pas intégré de manière innée, son intégration dépend du holding maternel. 22 Ce qu’il appelle « l’état d’unité » de l’enfant dépend de l’adéquation des soins maternels à ses besoins. Cela aboutit à une délimitation du dehors et du dedans indispensable à son individuation. « Puis il apparaît dans la suite du développement ce qu’on pourrait appeler une membrane de délimitation, qui se confond jusqu’à un certain point avec la surface de la peau ; elle se situe entre le « moi » et le « non-moi » de l’enfant. Ainsi le nourrisson en vient à avoir un intérieur et un extérieur et un schéma corporel.» Winnicott en 1960 (Cupa, 2006) L’intrication harmonieuse du corps et de l’esprit se fait grâce aux soins. La notion de « soin » est la clé de voûte du développement de l’enfant pour Winnicott. Les soins parentaux correspondent à la notion de « holding » (maintien), qui est la manière de contenir physiquement l’enfant comme le portage aux bras, le bercement et aussi le contenir psychiquement (disponibilité psychique pour son enfant, interpréter ses émotions...). Le « Handling » correspond aux soins du type laver, changer, habiller l’enfant. Cependant, la mère n'exécute pas ses différents actes de manière robotique, ils seront emprunts d’affectifs. Enfin « l’object presenting » est également une notion indispensable, ceci correspond à la manière dont la mère va présenter le monde à l’enfant. Elle va donc introduire l’extérieur à leur dyade mère-enfant. Des soins maternels satisfaisants édifient chez l’enfant un sentiment de continuité d’être. Au contraire, une carence des soins aboutit à une détresse extrême qui se traduit par le sentiment de partir en morceaux, le sentiment que la réalité extérieure ne peut servir à la réassurance. Un affaiblissement du moi en résulte. 23 « La santé mentale est donc le résultat des soins ininterrompus qui permettent une continuité du développement affectif personnel. » (Winnicott et al., 2011) d- Théorie de l’attachement La théorie de l’attachement de John Bowlby représente aussi un éclairage intéressant sur ce concept d’enveloppe psychique. Bowlby s’est intéressé aux conséquences des séparations précoces des enfants avec leurs parents. Ceci l’a conduit à élaborer la théorie de l’attachement. Cette théorie affirme que le bébé, de par son équipement inné, manifeste des conduites dirigées vers sa mère qui ont pour but le maintien le plus possible de la figure maternelle à côté de lui. Ces conduites de base sont la succion, le grasping (l’étreinte), les actions de poursuite (regard ou déplacements), les cris et le sourire. Ces conduites visent donc à garder la proximité physique avec la mère ou à faire revenir la mère à proximité. Cette proximité physique est indispensable pour que le bébé développe un sentiment de sécurité afin d’accroître ses potentialités psychiques et intellectuelles. Pour que le système d’attachement se déclenche, il faut se trouver dans une situation de détresse. En situation de stress et d’inconfort, le bébé interpelle alors sa figure d’attachement par les différentes conduites de base. Les réponses apportées par cette figure d’attachement sont très importantes dans la construction de la sécurité interne de l’enfant. 24 Le bébé est dépendant de la personne qui prend soin de lui, sans proximité avec cette personne, il est en danger de mort. Il va donc avoir sans cesse besoin d’aide pour grandir en toute sécurité. La sécurité émotionnelle de l’enfant va le rassurer pour ses explorations, ses expériences. On voit donc ici le lien entre les soins, réponses apportées par la figure maternelle avec la sécurité interne de l’enfant et sa capacité à faire des expériences sensori-motrices. Ces dernières étant fondamentales notamment pour créer des repères spatiaux (dedans/dehors, derrière/devant...). Ces repères enrichissent la conscience corporelle et les limites du corps. La théorie de l’attachement nous explique aussi comment les liens vont se construire, comment nous allons nous positionner dans nos relations, nos attentes, la confiance accordée aux autres, notre volonté à vouloir rencontrer de nouvelles personnes.... Un attachement insécure entraînera chez l'enfant une manière spécifique de s’adapter et de communiquer avec l’adulte. C’est Mary Ainsworth, dans les années 1960-1970 qui évoque les notions d’attachement « sécure » et « insécure ». En fonction de la qualité de soins reçue par notre « caregiver » ou figure d’attachement qui déterminera le type d’attachement. Les différents types d’attachement sont : ● Les attachements sécures ● Les attachements insécures o Les attachements anxieux-évitants o Les attachements anxieux-ambivalents ou résistants o Les attachements désorganisés 25 e- Syndrome d’hospitalisme Ce sont les travaux de Spitz, datant de 1946, qui mettent en évidence les conséquences chez les nourrissons suite à abandon ou leur privation de liens d’attachements stables. René Spitz observe 123 nourrissons âgés de 12 à 18 mois, de mère célibataire en prison. « Il décrit la « dépression anaclitique » (carence affective partielle) chez ces enfants séparés pendant le deuxième semestre de vie. Quand la séparation se prolonge, on observe une évolution vers un état de marasme, physique et psychique, que Spitz appelle « hospitalisme » (carence affective totale). »(Rousseau & Duverger, 2011) Plus récemment, c’est la pédiatre Emmi Pikler en 1975 qui remarque qu’un grand nombre d’enfants jeunes vivants en internat sont atteints du point de vue de leur développement de leur personnalité. Les travaux de E.Pikler et R.Spitz sur l’hospitalisme ont suscité une prise de conscience sur la manière de prendre en charge les enfants et l’importance de l’affectif dans la constitution de l’individu. Il est donc apparu le souci de prévenir l’apparition de troubles psychologiques liés aux carences affectives. Malheureusement cela reste un objectif non atteint, lors de ce stage en ITEP, j’observe de nombreux enfants placés en pouponnières et en foyer d’accueil. Une autre partie de ces enfants sont encore domiciliés chez leurs parents mais cet environnement constitue un milieu familial carencé voir maltraitant. Une grande partie des enfants rencontrés en prise en charge présente ce syndrome d’hospitalisme et/ou un type d’attachement insécure. Je clôturerai cette partie avec une étude récente qui montre encore l’actualité de ce problème. « Sur cinq années, entre 2005 et 2009, parmi la centaine d’enfants de moins de 3 ans admis à la pouponnière du Maine et Loire au titre de la protection de l’enfance, 40% présentaient un tableau gravissime au regard de leur état somatique, psychique ou de leur développement avec un fort risque de séquelles irréversibles. 15% présentaient des signes de souffrance psychiques sévères ou des troubles de la personnalité. »(Rousseau & Duverger, 2011) II- Le corps et ses représentations Un défaut de conscience corporelle est corrélé avec un défaut de connaissance de son schéma corporel et un trouble de l’image du corps. a- L’enveloppe corporelle Elle correspond à la peau. Elle nous limite, protège et contient le corps. L’enfant prend conscience de sa peau grâce à la relation du corps à corps. La peau est constituée de plusieurs couches : - L’épiderme, qui est la première couche, est elle-même constituée de quatre couches distinctes : la couche cornée, granuleuse, épineuse et basale. L’épiderme est renouvelé en moyenne tous les 60 jours. - Le derme, situé sous l’épiderme, est richement innervé, vascularisé et parcouru par beaucoup de vaisseaux lymphatiques. Il comporte de nombreux récepteurs sensoriels qui reçoivent les différents stimulus. - L’hypoderme est placé sous le derme et sert de barrière entre ce dernier et le reste du corps. La peau remplit des fonctions telles que la protection, la régulation thermique, l’absorption, l’excrétion et la production de vitamine D. La peau est le siège du toucher, elle est en étroite relation avec les autres organes des sens. Lors de sa naissance, l’enveloppe physique de l’enfant est visible. Cependant, l’enveloppe psychique est encore absente et va se construire au fil du temps, au fil des expériences. La notion de toucher, associée à la peau, est centrale et transversale. En effet, le système sensoriel humain se développe in-utero. À la naissance le bébé est doté de capacités sensorielles qui participent à son développement physique et psychique. Il passe d’un milieu aquatique à un milieu aérien qui est moins contenant. Le sentiment d’enveloppe diminue et le nouveau-né se retrouve confronté à des angoisses archaïques. Les interactions précoces et les sensations participent donc à l’édification de l’enveloppe psychocorporelle. Le toucher est le premier sens à apparaître et le dernier à se détériorer: on peut être muet, sourd, aveugle, on sent toujours un contact sur la peau. Les représentations mentales ne peuvent se construire qu’à partir de la sensation corporelle. Il est donc intéressant d’utiliser des médiations corporelles pour remettre cette connexion entre le sensoriel et ce qui est de l’ordre du symbolique. Ce sens du tactile est structurant pour le schéma corporel et unificateur grâce aux soins maternels. Il représente alors un véritable atout en psychomotricité. 28 b- Le corps ● Conception du corps en psychomotricité : Selon moi, il existe deux visions de pratiques psychomotrices. -La Rééducation Psychomotrice, de manière schématique elle vise à réduire un symptôme. Par exemple dans le cas d’une agitation motrice, d’un déficit de l’attention, on va viser à réduire le symptôme c’est-à-dire l’agitation. On pourra proposer des exercices permettant, de par leur répétition, à amener le sujet à avoir un meilleur contrôle de son attention, de ses troubles. -La Thérapie Psychomotrice, en reprenant l’exemple de l’instabilité, on ne visera pas directement à réduire les symptômes mais on va considérer ces derniers comme le signe d’une difficulté ou d’une souffrance qui existe en arrière-plan. On pourra proposer au patient des expériences lui permettant d’exprimer sa souffrance, de trouver des moyens d’expression qui lui permettent de dépasser sa souffrance. Ces deux visions de la pratique ne sont pas à opposer mais plutôt à combiner pour être au cœur de la psychomotricité. La question de comment on conçoit le corps renvoie à ce choix que peuvent faire certains psychomotricien(ne)s concernant leur pratique et impactera les moyens mis en œuvre ainsi que le sens de leurs actions. L’enfant, grâce à ses différentes expériences sensori-motrices, explore son corps. Puis, progressivement il pourra le nommer et s’en faire une représentation personnelle et unique. Les représentations de l’enfant seront dépendantes de ses diverses explorations qui seront aussi imprégnées de la dimension affective. 29 Tout cela est une construction progressive, des changements s’effectuent tout au long de notre vie. Par exemple, la formation au diplôme d’état de psychomotricien a provoqué chez un certain nombre d’étudiants des remaniements grâce à nos différentes médiations corporelles et au savoir théorique. Donc, toute notre vie nous pouvons repenser notre manière de percevoir et de se représenter notre corps. En Psychomotricité, le corps occupe une place centrale. Le corps est un outil indispensable pour le psychomotricien. En effet, notre spécialité sont les médiations corporelles. Le corps permet d’exprimer certaines souffrances et certains symptômes. Il nous permet de lire chez notre patient ce qu’il ne peut pas ou ne veut exprimer. Il est également notre outil médiateur, un moyen de rentrer en communication, un moyen d’expression. Le corps est aussi un outil d’apprentissage car l’expérience corporelle construit la conscience, la temporalité, la spatialité, la motricité... des notions que nous traitons en tant que psychomotricien. La richesse des expériences corporelles proposées aux enfants conditionne leur conscience. Le corps est l’instrument général de la compréhension du monde dans l’articulation du vécu corporel avec l’articulation psychique (en présence ou non du thérapeuthe). Donc, par le biais d’expériences corporelles nous pouvons aussi instaurer ou restaurer une conscience précaire. Je parlerai de ces deux no tions parfois séparément bien qu ’elles soient liées . En effet l’image du corps ne peut exister sans schéma corporel et inversement. Les représentations du corps (schéma corporel et image du corps) sont plastiques. Elles se réinventent, s’adaptent et se transforment en fonction de nos différents chemins de vie. C’est en cela que la plasticité de nos tissus donne une place importante à la psychomotricité. Historiquement, la notion de schéma corporel a été développée afin de répondre à des manifestations pathologiques du système nerveux. Le schéma corporel se construit en parallèle de la maturation du système nerveux. P. Bonnier, dès 1833, introduit l’idée d’une représentation topographique du corps dont les altérations pathologiques déterminent des troubles particuliers (tel qu’une perte de la perception topographique du corps ou une confusion des différentes régions du corps). En 1895, il donne le nom de schéma corporel, il le définit comme une représentation permanente, une figuration spatiale du corps. En 1908, Pick propose la notion « d’image spatiale du corps » établissant une relation entre les sensations perçues dans le corps et les représentations de celui-ci, relation qui permet d’estimer la localisation des stimulations. schéma corporel constituerait une interface permettant une relation adaptée aux sensations et à l’environnement. Enfin Head, dans les années 1920, parle de « modèle postural du corps ». Il ajoute la dimension temporelle et donc le caractère plastique et en perpétuel construction. 31 D’après Julian de Ajuriaguerra (1970) le schéma corporel est « édifié sur la base des impressions tactiles, kinesthésiques, labyrinthiques et visuelles, le schéma corporel réalise dans une construction active constamment remaniée des données actuelles et du passé, la synthèse dynamique, qui fournit à nos actes, comme à nos perceptions, le cadre spatial de référence où ils prennent leur signification » (Scialom et al., 2011) p.216 D’après Jean Michel Albaret « le schéma corporel (système de localisation) correspond à un ensemble de représentations pour l’action et répond donc à la fameuse question « Où? » (Où sont les membres et où est la cible de l’action) mais aussi à la question « Comment? » (Comment utiliser l’ensemble du corps pour réaliser le but fixé). Il fournit les informations sur la posture et les coordonnées spatiales des différentes parties du corps qui détermine aussi ce qu’on appelle le référentiel égocentrique. Il intervient donc pour la locomotion et la préhension. »(Scialom et al., 2011) p.216 De nombreux auteurs ont défini les différentes étapes du développement du schéma corporel. Voici une proposition de ces différentes étapes : Le corps subi (0-3mois) : La motricité à ce stade reste de type « sous corticale » donc on parle de « motricité réflexe ». C’est le stade de l’impulsivité motrice, les gestes sont explosifs, non orientés. Il a une conscience de son corps que fragmentaire et peu différenciée de l’autre. Par la maturation du système nerveux (qui suit les lois céphalo-caudales et proximaux-distales) et la stimulation de l’environnement lui apportant sécurité physique et affective, l’enfant va passer peu à peu de cette motricité réflexe à une motricité plus libérée et ainsi volontaire. Le corps vécu (3 mois- 3ans) : Freinet parle d'un « tâtonnement expérimental » lorsqu'un résultat nouveau est obtenu, parfois par hasard, l'enfant cherche à le reproduire avec des variantes. L'enfant est alors capable d'utiliser un schéma intériorisé pour une situation nouvelle. Donc par la répétition des multiples expériences sensorielles et motrices, l’enfant va de mieux en mieux maîtriser son corps. Par l'expérience vécue du moment, l'enfant délimite progressivement son « corps » du monde des objets. S'établit alors une première ébauche des représentations corporelles en tant que représentations conscientes. L’adulte met aussi les mots sur les sensations de l’enfant, ce qui permet le passage de la sensation à la perception. A ce stade apparaissent les premiers apprentissages et connaissances corporelles. C’est également la période sensori-motrice, selon Piaget, elle correspond au développement et à la coordination des capacités sensorielles et motrices du bébé. L’intelligence est liée à l’action, le bébé touche les objets et expérimente, peu à peu il comprend le fonctionnement de son corps et commence à le différencier des objets. Le corps perçu (3 ans -7 ans) : La motricité continue de s’affiner. L’enfant passe d’une connaissance globale et d’une perception de son corps à une connaissance topologique des différentes parties du corps. A cet âge, l’enfant acquiert le jeu symbolique et la latéralité se met en . Tout ceci contribue à l’acquisition des repères spatiaux et temporels. Il y a une meilleure connaissance des sensations éprouvées, de la conscience du corps dans l’espace et un meilleur ajustement postural. 33 Le corps représenté (7 ans-12 ans) : L’enfant passe d’une connaissance globale à une connaissance topologique des différentes parties de son corps : il peut nommer et situer ces différentes parties. La représentation mentale du corps se constitue à cette période. Il peut ainsi mieux représenter ce corps au travers du dessin. Il acquiert également l’efficience motrice c’est-à-dire qu’il peut s’ajuster toniquement, exécuter des mouvements plus précis, anticiper... L’enfant prend donc conscience de son corps en mouvement. Le schéma corporel va ainsi se construire via toutes les informations issues du corps afin de constituer un schéma de référence. Il se construit sur l’expérience sensori-motrice et grâce à l’aide de l’environnement. En effet, il se structure grâce à la maturation neurologique, l’équipement sensoriel et la motricité mais également grâce à l’environnement humain et le dialogue tonico-émotionnel. Il a donc un lien avec l’affectif. Il regroupe la connaissance et la conscience que l’on a de son corps. Sa structuration repose sur une multitude d’informations neurosensorielles : les sensations tactiles, visuelles et vestibulaires. La synthèse de toutes ces informations sensorielles permet à chaque instant d’établir une représentation unifiée de notre corps et fournit un repère de base pour s’organiser. En effet, la conscience d’un schéma de référence permet de s’orienter et de déterminer un haut et un bas, une droite et une gauche, un devant et un derrière... Son intégration aura des répercussions sur l’ensemble du développement de l’enfant comme son aisance motrice et gestuelle, se repérer dans l’espace et le temps. 34 A l’image de la psychomotricité, le schéma corporel n’est donc pas à considérer comme un concept uniquement neurologique et objectif, mais s’articule également avec la dimension de l’expérience vécue subjective. Le développement psychomoteur s’appuie sur la boucle sensation-perception-représentation : les sensations, une fois intégrées, nourrissent des représentations, des images mentales qui ouvrent le champ à d’autres sensations. C’est cette boucle qui est mise en jeu tout au long des différents stades nécessaires à la construction du schéma corporel. d- L’ image du corps Cette notion est issue de la psychanalyse. À partir de Bergson (1896), l’image du corps a beaucoup été l’objet de la psychanalyse. Il définit l’image du corps comme la perception actuelle que le sujet a de son corps, et d’autre part, le produit de l’assimilation des expériences passées qui lui ont conféré́ son unité́. Pour lui, l'image du corps est à la fois consciente et inconsciente. Pour Bergson, il y a donc pour lui une mémoire du corps (celle des expériences passées) et l'image du corps est garante de l'identité du sujet qui incarne la continuité́ du rapport que l’individu établit entre le passé et le présent. D’après Jean Michel Albaret « L’image du corps (système de perception des formes) sert à l’identification et à la reconnaissance du corps et répond à la question « Quoi? » (Quel corps suis-je? Quelles sont les particularités de mon corps ou de mon apparence corporelle?). Elle regroupe des perceptions et des attitudes à l’égard de son corps propre qui s’expriment par des pensées, des convictions, des sentiments mais aussi des comportements. Cette image du corps est influencée par un ensemble de facteurs socioculturels, psychologiques, biologiques. » (Scialom et al., 2011) p.216 35 Gallagher définit l’image du corps comme un « système de perception, d’attitudes et de croyances se rapportant à son propre corps. »(Scialom et al., 2011) L’image du corps présente un rôle social important car elle se construit à travers la relation à autrui. Elle est unique, propre à chacun et liée à nos différentes expériences affectives, sensorielles, sociales... L’image du corps est à l'origine du narcissisme du sujet et de sa subjectivité. Elle assure une assise identitaire. Il permet au « Moi » de se constituer par rapport à son environnement. En effet, F. Dolto l’assimile à la « trace structurale de l’histoire émotionnelle d’un être humain ».(Scialom et al., 2011) Grâce à l’image du corps nous sommes ce que nous sommes. En relaxation, on travaille autour de l’image du corps. J. Lacan propose le « stade du miroir ». Celui-ci permet à l’enfant d’acquérir une image corporel unifiée ainsi que la représentation de son propre corps. L’image du corps se constitue à partir d’images qui sont d’abord morcelées et qui vont petit à petit se rassembler autour d’une expérience : l’expérience du miroir. L’enfant va s’identifier progressivement à l’image qu’il voit dans le miroir. Au début, l’enfant prend son image pour un autre réel. Par la suite, il va se rendre compte que ce qu’il voit dans le miroir est une image et non une autre personne. Enfin, l’enfant s’identifie à l’image du miroir en reconnaissant cette image comme étant la sienne. stade est alors constructif pour le développement du « moi », il acquiert alors son image du corps comme un tout unifié et différent d’autrui. Ce qui aboutit à la constitution de son identité.	14,551
https://openalex.org/W3155089635	OpenAlex	Open Science	CC-By	2,021	Keeping in time with social and non-social stimuli: Synchronisation with auditory, visual, and audio-visual cues	Juliane J. Honisch	English	Spoken	9,939	17,635	Juliane J. Honisch1, Prasannajeet Mane1, Ofer Golan2 & Bhismadev Chakrabarti1 Juliane J. Honisch1, Prasannajeet Mane1, Ofer Golan2 & Bhismadev Chakrabarti1 Everyday social interactions require us to closely monitor, predict, and synchronise our movements with those of an interacting partner. Experimental studies of social synchrony typically examine the social-cognitive outcomes associated with synchrony, such as affiliation. On the other hand, research on the sensorimotor aspects of synchronisation generally uses non-social stimuli (e.g. a moving dot). To date, the differences in sensorimotor aspects of synchronisation to social compared to non-social stimuli remain largely unknown. The present study aims to address this gap using a verbal response paradigm where participants were asked to synchronise a ‘ba’ response in time with social and non- social stimuli, which were presented auditorily, visually, or audio-visually combined. For social stimuli a video/audio recording of an actor performing the same verbal ‘ba’ response was presented, whereas for non-social stimuli a moving dot, an auditory metronome or both combined were presented. The impact of autistic traits on participants’ synchronisation performance was examined using the Autism Spectrum Quotient (AQ). Our results revealed more accurate synchronisation for social compared to non-social stimuli, suggesting that greater familiarity with and motivation in attending to social stimuli may enhance our ability to better predict and synchronise with them. Individuals with fewer autistic traits demonstrated greater social learning, as indexed through an improvement in synchronisation performance to social vs non-social stimuli across the experiment. Everyday social interactions such as talking with friends, playing games (i.e. pat a cake) or lifting a table together require us to continuously monitor and predict the behaviour of others, and adjust our movements accordingly to achieve smooth interactions. The ability to closely synchronise our movements with external stimuli has been proposed to have evolutionary functions for resource acquisition and reproduction1 by facilitating cooperation2 and social cohesion3,4. One of the most common channels used for synchronous interactions is the verbal channel, as in simple parent–child games, singing, sloganeering or through the entrainment of speech rhythms during everyday conversations5,6.i y y Previous research on sensorimotor synchronisation has largely used finger tapping paradigms to examine synchronous responses to non-social stimuli, such as flashing lights or auditory metronomes7,8. However, most stimuli that we synchronise to are social in nature, e.g. the movements of another person, dance, speech or songs. Studies that have investigated synchronising to social stimuli have generally used a different class of paradigms with more ecological validity, e.g. www.nature.com/scientificreports www.nature.com/scientificreports Scientific Reports \| (2021) 11:8805 Juliane J. Honisch1, Prasannajeet Mane1, Ofer Golan2 & Bhismadev Chakrabarti1 synchronous arm curling9, walking together10, dance-like movements11 or synchronous bouncing12. The focus of these studies has been on social affiliative and cognitive outcomes rather than the accuracy of synchronous responses to social stimuli. Due to this historical diversity of experimental approaches in studying synchrony in social and non-social domains, the differences in synchronising to social vs non-social stimuli remain largely unknown. While social compared to non-social stimuli have been shown to be processed differently in multiple studies13–17, none of these studies have used synchrony based measures to examine how we physically interact with such stimuli. Most studies in this area use paradigms where observ- ers passively observe social and non-social pictures/videos presented on a screen14,16–19 . In contrast, interper- sonal synchrony involves an active involvement with the environment, and is more typical of real-world social situations where participants are not merely passive observers. 1Centre for Autism, School of Psychology and Clinical Language Sciences, University of Reading, Reading RG6 6AL, UK. 2Department of Psychology, Bar-Ilan University, Ramat Gan, Israel. email: J.J.Honisch@reading.ac.uk; b.chakrabarti@reading.ac.uk 1Centre for Autism, School of Psychology and Clinical Language Sciences, University of Reading, Reading RG6 6AL, UK. 2Department of Psychology, Bar-Ilan University, Ramat Gan, Israel. email: J.J.Honisch@reading.ac.uk; b.chakrabarti@reading.ac.uk Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ Table 1. Parameter estimates for the fixed effect omnibus tests. Fixed effect omnibus tests F Num df Den df p Condition 16.697 2 2590.7 < .001 Stimulus type 797.117 1 2604.7 < .001 AQ 0.166 1 40.1 0.686 Gender 0.307 1 39.9 0.583 Condition * Stimulus Type 44.432 2 2592.6 < .001 Table 1. Parameter estimates for the fixed effect omnibus tests. The real world rarely offers stimuli in a single sensory modality. Accordingly, it is vital to consider cues in single as well as multiple sensory modalities within each class of stimuli (social and non-social). Previous reports using non-social stimuli have shown closer synchronisation performance in terms of mean asynchrony when synchronising to an auditory compared to a visual metronome20–22. This finding is not unexpected, as audi- tion has been found to be superior in temporal processing, whereas the visual modality is superior in spatial processing20. However, visual dynamic stimuli compared to discrete visual stimuli have been found to improve synchronisation performance23. Specifically, in bimodal (audio-visual) stimuli, individuals have been shown to optimally integrate information from both modalities24. Juliane J. Honisch1, Prasannajeet Mane1, Ofer Golan2 & Bhismadev Chakrabarti1 Elliott, Wing and Welchman25 showed that synchronous finger tapping with bimodal discrete stimuli was more accurate compared with unimodal stimuli. However, little is known about the impact of the number of modalities on synchronisation to social stimuli. p y Engaging in synchronous interactions facilitates social bonds, by increasing liking3, trust26, prosocial behaviour27,28 and reducing outgroup bias29. Individuals diagnosed with autism spectrum disorders (ASD) typically face challenges in social communication and sensory processing30, and often experience difficulties in integrating multisensory information31. In lab-based studies, autistic individuals show a reduced preference for social compared to non-social stimuli32–34. ASD has also been associated with atypical spontaneous facial mim- icry and spontaneous motor synchronisation with another person35–38. In comparison with typically developing children and adults, individuals with ASD have been found to produce weaker and more variable synchronisation behaviours to social stimuli39,40.hf The present study aims to systematically examine the differences in synchronisation behaviour to social compared to non-social stimuli. Here, we developed a novel verbal response paradigm in which participants syn- chronised their verbal response in time with social or non-social stimuli (presented unimodally, audio or visual) or bimodally (audio-visual). We predict that individuals will perform differently in how well they synchronise with social compared to non-social stimuli. One possibility is that individuals will be worse in synchronising to social stimuli due to their greater complexity (i.e., a face is significantly more visually complex compared to a dot). Another possibility is for individuals to show better performance in synchronizing with social stimuli, due to the greater reward value typically attributed to social stimuli16,19. The second possibility is supported by empirical and theoretical accounts suggesting greater reward response associated with motor alignment41–43. For both social and non-social stimuli, synchronisation with audio-visual combined conditions is expected to result in lower asynchrony compared to the audio and visual only conditions25. Further, in line with the literature on auditory superiority when synchronising with a metronome, we predict closer synchronisation with auditory compared with visual stimuli20–22. p In this study, autistic traits were measured using the Autism Spectrum Quotient44 (AQ). Autistic traits are distributed continuously throughout the population with similar aetiology at both ends, allowing us to examine the impact of autism-related variation at a population level45. We predict that autistic traits will be negatively associated with synchronisation performances across tasks, in line with earlier findings of weaker and more variable synchronisation performances in individuals with ASD 38–40. www.nature.com/scientificreports/ www.nature.com/scientificreports/ Table 2. Parameter estimates for the fixed effects of the model. Conditions are denoted as follows (A: Auditory, C: Combined, V: Visual). Stimulus Types are denoted as Social or Nonsocial. Fixed effects parameter estimates Names Effect Estimate SE 95% Confidence Interval df t p Lower Upper (Intercept) (Intercept) 0.34766 0.0180 0.31248 0.38285 55.0 19.368 < .001 Condition1 C–A − 0.01649 0.0111 − 0.03831 0.00533 2595.1 − 1.481 0.139 Condition2 V–A − 0.03573 0.0113 − 0.05788 − 0.01357 2600.9 − 3.161 0.002 Stimulus type 1 Social–Nonsocial − 0.24283 0.0117 − 0.26571 − 0.21995 2592.0 − 20.804 < .001 AQ AQ 3.96e−4 9.74e−4 − 0.00151 0.00231 40.1 0.407 0.686 Gender 1 Male-Female − 0.00565 0.0102 − 0.02564 0.01433 39.9 − 0.554 0.583 Condition 1 * Stimulus type 1 C–A * Social-Nonsocial 0.01112 0.0164 − 0.02111 0.04336 2587.5 0.677 0.499 Condition 2 * Stimulus type 1 V–A* Social-Nonsocial 0.14213 0.0167 0.10937 0.17488 2594.0 8.505 < .001 Table 2. Parameter estimates for the fixed effects of the model. Conditions are denoted as follows (A A dit C C bi d V Vi l) Sti l T d t d S i l N i l Table 2. Parameter estimates for the fixed effects of the model. Conditions are denoted as follows (A: Auditory, C: Combined, V: Visual). Stimulus Types are denoted as Social or Nonsocial. hif Figure 1. This figure shows the effect of Stimulus Type (Social, Non-social) and Condition (A: Auditory, C: Combined, V: Visual) on mean asynchrony in seconds. The error bars signify a 95% confidence interval. Figure 1. This figure shows the effect of Stimulus Type (Social, Non-social) and Condition (A: Auditory, C: Combined, V: Visual) on mean asynchrony in seconds. The error bars signify a 95% confidence interval. comparison to visual conditions [tvisual-audiovisual(2584) = 9.156, p < 0.001; tauditory-visual(2584) = − 8.644, p < 0.001]. A full list of post-hoc comparisons is provided in the Supplementary Materials.f f It is possible to measure synchrony using either the visual or auditory onsets in response to the combined audio-visual stimulus. Synchronisation performance to social combined (audio-visual) stimuli was further exam- ined to test if it differed as a function of using auditory vs visual onsets to define the target cue. This analysis was restricted to social stimuli alone, since auditory and visual onsets for non-social audio-visual stimuli were programmed to be identical. if Auditory, C: Combined, V: Visual). Stimulus Types are denoted as Social or Nonsocial. Resultsi Model fit statistics for the linear mixed model implemented were as follows: AIC = −1726.84, BIC = −1602.964, R-squared conditional = 0.273 AIC = −1726.84, BIC = −1602.964, R-squared conditional = 0.273 Results of the linear mixed model analysis are presented below in Tables 1 and 2.hf y p The model revealed a main effect of Stimulus Type (F(1,2604.7) = 797.12, p < 0.001) with social stimuli associated with significantly lower mean absolute asynchrony (estimated marginal mean = 142 ms, 95%CI [129, 154]) compared to non-social stimuli (estimated marginal mean = 334 ms, 95%CI [322, 345]). A main effect of Condition was also observed (F(2,2590.7) = 16.70, p < 0.001) with visual stimuli (estimated marginal mean = 265 ms, 95%CI[251, 279]) associated with significantly larger absolute mean asynchrony compared with auditory (estimated marginal mean = 230 ms, 95%CI[216, 243]) and audio-visual combined stimuli (estimated marginal mean = 219 ms, 95%CI[216, 243], see Fig. 1). No significant difference between auditory and audio- visual combined conditions was observed. An interaction effect between condition and stimulus type was noted (F(2,2592.6) = 44.43, p < 0.001). All pairwise post-hoc comparisons show that social stimuli were associated with better synchronization performance compared to non-social stimuli, irrespective of sensory modality. For non- social stimuli, participants synchronized better with visual than with auditory conditions [t(2600) = 3.160, p = 0.023]. For social stimuli, synchronisation performance was better for auditory and audio-visual conditions in https://doi.org/10.1038/s41598-021-88112-y Scientific Reports \| (2021) 11:8805 \| Table 2. Parameter estimates for the fixed effects of the model. Conditions are denoted as follows (A: www.nature.com/scientificreports/ The results revealed a significant difference in mean asynchrony as a function of onset type [t(2282) = − 10.94, pBonferroni < 0.001], with higher mean asynchrony associated with the visual onset (estimated marginal mean = 201 ms, 95% CI [264.1,320]) than with the auditory onset (estimated marginal mean = 142 ms, 95% CI [75.4,130]).f An exploratory analysis to measure learning/practice effects through the experiment revealed a main effect of trial [F(1,2586.2) = 9.942, p = 0.002], a significant two-way interaction of trial and stimulus type [F(1,2586.2) = 7.341, p = 0.007] as well as a significant three-way interaction between stimulus type, trial, and AQ [F(2,2588.3) = 3.154, p = 0.043] (see Figs. 2 and 3). https://doi.org/10.1038/s41598-021-88112-y Scientific Reports \| (2021) 11:8805 \| www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 2. This figure shows the two-way interaction for the learning effect, indicating a decrease in mean absolute asynchrony for social but not for non-social conditions. Stimulus Type (Social, Non-social) on mean asynchrony in seconds. The error bars signify a 95% confidence interval. Figure 3. This figure shows the three-way interaction for the learning effect and AQ, indicating a decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. The left panel figure illustrates that participants who scored 1 SD below the sample mean showed a steep decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. In contrast, participants who scored 1 SD above the mean AQ scores (right panel figure) showed a less steep decrease in mean absolute asynchrony in response to social vs non-social stimuli across trials. Figure 2. This figure shows the two-way interaction for the learning effect, indicating a decrease in mean absolute asynchrony for social but not for non-social conditions. Stimulus Type (Social, Non-social) on mean asynchrony in seconds. The error bars signify a 95% confidence interval. Figure 2. This figure shows the two-way interaction for the learning effect, indicating a decrease in mean absolute asynchrony for social but not for non-social conditions. Stimulus Type (Social, Non-social) on mean asynchrony in seconds. The error bars signify a 95% confidence interval. Figure 3. This figure shows the three-way interaction for the learning effect and AQ, indicating a decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. www.nature.com/scientificreports/ The left panel figure illustrates that participants who scored 1 SD below the sample mean showed a steep decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. In contrast, participants who scored 1 SD above the mean AQ scores (right panel figure) showed a less steep decrease in mean absolute asynchrony in response to social vs non-social stimuli across trials. Figure 3. This figure shows the three-way interaction for the learning effect and AQ, indicating a decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. The left panel figure illustrates that participants who scored 1 SD below the sample mean showed a steep decrease in mean absolute asynchrony across trials for social compared with non-social stimuli. In contrast, participants who scored 1 SD above the mean AQ scores (right panel figure) showed a less steep decrease in mean absolute asynchrony in response to social vs non-social stimuli across trials. Figure 2 illustrates the data from the two-way interaction by showing a monotonically decreasing trend of mean absolute asynchrony for social trials, suggesting the existence of a practice/learning effect. This trend is not seen in response to the non-social trials. Figure 3 demonstrates the three-way interaction between stimulus type, trial, and AQ. Figure 2 illustrates the data from the two-way interaction by showing a monotonically decreasing trend of mean absolute asynchrony for social trials, suggesting the existence of a practice/learning effect. This trend is not seen in response to the non-social trials. Figure 3 demonstrates the three-way interaction between stimulus type, trial, and AQ. www.nature.com/scientificreports/ This observation is consistent with a recent report showing reduced integration of social information in a learning task in individuals with high autistic traits55.hf g g g g The observation of differences in response to social and non-social stimuli raises an important question about the origin of these differences. The low-level properties of social and non-social stimuli used in this study are different (e.g., contrast, colour, nature of sound). It is possible that the distinction between ‘social’ and ‘non- social’ stimuli is a cumulative effect driven by a large number of such low-level properties. Perfect matching for all stimulus properties will render the two sets of stimuli identical to one another. It is worth noting that the category of ‘social’ stimuli represents a circumscribed set of low-level stimulus features (e.g., flesh tones for visual stimuli, sound within the vocal frequency range for auditory stimuli). Whether there is a ‘social’ advantage over and above all potential physical characteristics of the stimulus remains an open question. Although our paradigm has higher ecological validity by presenting a real recording of a human partner, examining synchronisation that replicates a real-life social interaction that involves mutual and reciprocal adaptation would be of interest for future research.i In view of the significant interaction between stimulus type and condition observed in the main analysis, we separately analysed synchronisation performance by stimulus type. For non-social stimuli, there was little difference between the three conditions (see Fig. 1), but participants synchronised better with visual, than with auditory stimuli. This finding is in contrast to a previous study that showed better synchronization of finger tap responses to an auditory compared to a visual stimulus56. One potential explanation is that continuous visual stimuli provide a more salient temporal cue to allow for better temporal judgements than discrete auditory stimuli57. The continuity of visual stimuli like the ones used in this study provide sufficient time to the participant to prepare a response to synchronize with the target cue58,59. Synchronization performance with the combined audio-visual stimulus was not significantly different from that with either of the unimodal stimuli.i gi yf For social stimuli, an opposite pattern was observed—with participants synchronising significantly better with auditory than visual stimuli. This pattern of results is consistent with the observation that the auditory modality performs better than vision in tasks that involve temporal processing60,61. www.nature.com/scientificreports/ attention to social stimuli such as human sounds, movements and facial features47,48. Increased reward value of target stimuli can influence both the starting point as well as the rate of sensory data accumulation49,50. Greater relative reward value for social stimuli can therefore potentially explain closer synchronous performances for such stimuli as observed in our current study. Another plausible explanation for this difference is the use of verbal responses in the present study. Verbal responses match the nature of the social but not the non-social stimuli in our paradigm. The compatibility between the action performed in the social stimuli and the motor response performed by participants may have improved motor simulation by activating the same neural systems involved in perceiving and producing the same motor response51. Consequently, participants’ predictive model of the stimuli, along with participants’ motor planning and execution could have been enhanced, resulting in closer synchronous performances52. Further, given the social nature of human actions, whether it is making a ba sound or a finger tap, it is not entirely possible to decouple response compatibility from the sociality of stimuli. Another possible contributing factor for the improved synchronisation observed for social stimuli could be the lower pitch of the male and female actor in comparison with the higher pitch of the metronome. Synchronisation with auditory metronomes can be influenced by the pitch of the sounds, with better synchronisation associated with lower pitch sounds53,54. However, no systematic analysis has been conducted to estimate the magnitude of this effect on a wide range of pitches that are comparable to those used in this study, and whether this finding can be generalised to human vocal sounds.fif g Our exploratory analysis to check for practice/learning effects revealed a significant difference between the social and non-social conditions. Across the whole sample, individuals tended to perform better (i.e., with less absolute mean asynchrony) as the experiment progressed. This learning effect was greater for social compared to non-social stimuli, which could reflect greater attention being drawn to the dynamic social stimulus (a face saying ‘ba ba’) in comparison to the bouncing dot. This social learning effect was greater in individuals with lower autistic traits, than those with higher autistic traits. Discussionh The present study systematically tested the differences in synchronising with social and non-social stimuli, when presented in single or multiple sensory modalities. Mean asynchrony for trials in response to non-social stimuli was found to be greater than in response to social stimuli. Additionally, mean absolute asynchrony for visual stimuli were found to be higher than for auditory and audio-visual (combined) stimuli. No effect of autistic traits or gender were noted on mean absolute asynchrony.i Participants were significantly better in synchronising with a social compared to non-social stimuli, across all three conditions (auditory/visual/audio-visual), despite the greater variability in the target cue timings for social stimuli. This finding could potentially be an outcome of increased attention or reward value ascribed to social over non-social stimuli17,19,34,46. Infants less than four weeks old have been found to show preferential Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ Materials and methods Participants and design. Fifty-three psychology undergraduates (29 females, 24 males; Mage = 21.01 years, range = 18–31 years) took part in the study in exchange for course credit or for cash and were screened for photosensitive epilepsy. The study had a 2 (stimulus type: social, non-social) × 3 (condition: audio, visual, audio- visual combined) within-subject design. Sample size was determined a priori using GPower 3.164. The analysis was based on an effect size calculated from a previously published study demonstrating improved synchronous performance to auditory compared to visual cues25 (d = 1.3). However, since our response modality as well as stimuli type differed from the study above, we chose a more conservative effect size (d = 0.7). The analysis sug- gested that the minimum acceptable total sample size needed to achieve a power of .80 was 41. Ethics. The study was approved by the School of Psychology and Clinical Language Sciences Ethics Com- mittee at the University of Reading. The experiment was performed in accordance with the relevant guidelines and regulations, and participants provided informed, written consent. Written informed consent was obtained to publish identifying images of the social stimulus, in an online open access publication. Experiment setup and apparatus. Participants synchronised their verbal responses (a ‘ba’ sound) to either audio, visual or audio-visual combined target cues. A SparkFun sound detector and an acquisition hard- ware (National Instruments NI-USB 6343) were used to record the presence of participants’ verbal responses. One computer was used to control and present both the visual and auditory stimuli (target cues) on the screen.h p p y g Two types of target cues were presented, social and non-social (see Fig. 4). The social cues consisted of video recordings of both a male and a female actor performing a rhythmical ‘ba’ sound. The gender of the actor in the video recordings was matched with the gender of the participant, controlling for gender effects during synchronous activities65.h y The actor was presented from the collar bone upwards, with a neutral facial expression, a controlled back- ground, and wearing fitted black clothes. For visual social cue conditions, the video recording was presented without sound. Here the opening of the actor’s mouth was the target cue. For audio social conditions, a blank black screen with a white fixation cross was presented with the audio recording of the actor’s rhythmical ‘ba’ sound. www.nature.com/scientificreports/ One possibility is that this pattern of auditory dominance in synchronisation tasks is observed once the mismatch between the target stimulus and response modality is minimised. For finger tapping tasks such as the one by Hove and others62, the visual stimulus of a moving bar was similar to the response modality (a moving finger). Once this mismatch is minimised, the cognitive efforts could be directed entirely to the temporal aspects of the stimuli, which would result in an audi- tory dominance effect. In the current study, the mismatch between the target stimulus and the response modality is significantly lower for the social stimuli than the non-social stimuli, which leads to the expected pattern of auditory dominance for the social stimuli. For the main analysis, all target cues were identified auditorily, i.e. the time of peak of every ‘ba’ utterance was used to calculate the asynchrony. When visual onsets (first frame of mouth opening) were used to calculate the asynchrony, an identical pattern was observed for synchronisation with unimodal auditory and visual stimuli. Interestingly however, the synchronisation performance with the combined audio-visual stimulus differed significantly as a function of which onset type was used. When audi- tory onsets were used, synchronisation performance was similar to the unimodal auditory condition. However, when the visual onsets were used, performance was closer to, but significantly worse than the unimodal visual condition. This result suggests that participants tend to synchronise their verbal responses to the auditory rather than the visual cue in the audio-visual condition.f Contrary to our original hypothesis, no effect of autistic traits was noted in relation to synchronisation performance in response to either social or non-social stimuli per se. While this result is consistent with a recent report showing no autistic deficit in auditory-motor synchronisation using a finger tapping task63, it is in contrast to another report using coherence as a measure of synchronization of bodily movements between a live experimenter and the participant40. A direct comparison of the current results with these previous studies is not straightforward due to the different nature of stimuli and response modalities used. We note, however, Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ Figure 4. An illustration of the experimental design. Each stimulus type (social or non-social), contained audio only, visual only and audio-visual combined stimuli. www.nature.com/scientificreports/ For social stimuli, the audio condition consisted of a recording of a rhythmical verbal ‘ba’ response (either female or male), the visual condition consisted of a video recording of a female or male person performing the rhythmical ‘ba’ response without the presentation of the sound, and the combined condition consisted of a video recording of the female or male person performing the rhythmical ‘ba’ response with the sound. For non-social stimuli, either a rhythmical metronome beep (audio), a white vertically moving dot (visual) or both the metronome beep coinciding with the moving dot (combined) were presented. Figure 4. An illustration of the experimental design. Each stimulus type (social or non-social), contained audio only, visual only and audio-visual combined stimuli. For social stimuli, the audio condition consisted of a recording of a rhythmical verbal ‘ba’ response (either female or male), the visual condition consisted of a video recording of a female or male person performing the rhythmical ‘ba’ response without the presentation of the sound, and the combined condition consisted of a video recording of the female or male person performing the rhythmical ‘ba’ response with the sound. For non-social stimuli, either a rhythmical metronome beep (audio), a white vertically moving dot (visual) or both the metronome beep coinciding with the moving dot (combined) were presented. that consistent with a previous report, a weaker social learning effect was observed in individuals with higher autistic traits55.i In summary, our findings suggest that humans synchronise their responses more closely with social compared with non-social stimuli. This ‘social advantage’ is likely to be driven by the preferential attention and reward linked to perceiving and interacting with other humans. Potential future research could formally examine the dependence of these results on the response modality (verbal response/ finger tapping), as well as test the impact of attention given to social compared to non-social stimuli on synchronisation performance. In summary, our findings suggest that humans synchronise their responses more closely with social compared with non-social stimuli. This ‘social advantage’ is likely to be driven by the preferential attention and reward linked to perceiving and interacting with other humans. Potential future research could formally examine the dependence of these results on the response modality (verbal response/ finger tapping), as well as test the impact of attention given to social compared to non-social stimuli on synchronisation performance. www.nature.com/scientificreports/ The AQ has 50 items measuring diverse dimen- sions of the autistic phenotype, such as, “I enjoy meeting new people”. Participants rate their level of agreement with each statement on a 4-point Likert scale ranging from ‘definitely agree’ to ‘definitely disagree’. Ratings are then collapsed to a yes/no scoring. Thus, the AQ scores range from 0 to 50, with autistic individuals typically scoring higher than neurotypicals. In the present study an online version of the questionnaire was administered. Task and procedure. Participants were asked to attend two experimental sessions, each lasting around 50 minutes, with a minimum gap of one day from one another. Each session contained either the social or non- social stimuli. The order of these sessions was counterbalanced across participants. Participants completed the AQ online when signing up for this study. For the experimental sessions, participants arrived at the laboratory individually, were greeted by the experimenter, and were seated at a table facing a PC monitor. Participants learned that the goal of the experiment was to examine the effects of different types of stimuli on their abil- ity to verbally synchronise with them. Once participants read the information, consent was provided. In the social-stimuli session, female participants were shown the video and audio recordings of the female actor, while male participants were shown the recordings of the male actor. For visual social conditions, participants were instructed to produce a ‘ba’ sound in synchrony with the mouth opening of the actor presented in the video. In auditory social conditions, participants were asked to synchronise their ‘ba’ response in time with the ‘ba’ sound of the actor. For audio-visual social conditions, participants were instructed to synchronise their responses to both the audio-visual cue (‘ba’ sound and mouth opening) of the actor presented in the video. In the non-social stimuli session, both male and female participants were either asked to produce a ‘ba’ sound in synchrony with computer˗generated audio and visual stimuli. For non-social auditory conditions, participants synchronised their ‘ba’ response in time with an auditory metronome, whereas for non-social visual conditions participants synchronised their responses with a moving white dot at the lowest point on the vertical axis. Finally, for non- social audio-visual conditions, participants were instructed to synchronise their ‘ba’ responses with both the metronome and the moving dot simultaneously. The duration of each experimental session was 40 minutes. Analysis. www.nature.com/scientificreports/ Non-social cues consisted of inanimate stimuli. For visual non-social condition, a white dot (2.54 cm diameter) was presented on a black background. The refresh rate of the PC monitor was 60 Hz. The dot was moving vertically with a fixed amplitude of 20 cm, using a pre-generated sine wave function. The lowest point of the downwards motion was the target cue. In the auditory non-social conditions, the generated trajectory of the dot movement was used to estimate the corresponding non-social auditory cue. The lowest peak on the x-axis for each downwards oscillation was used to generate a series of rhythmical metronome tones with a tone duration of 50 ms and at 700 Hz. In audio-visual non-social conditions, both the dot motion with its corresponding audio metronome tones were presented simultaneously (see Supplementary Material for stimulus generation code). Although there could be small discrepancies under the 10-ms range between the programmed and presented audio/visual onsets for the non-social stimuli66, we minimised the risk of such discrepancies by using a power- ful graphics card (NVIDIA GTX 650, 4 GB), and a significantly higher screen refresh rate than the stimulus frequency. However, it should be noted that in the absence of an external photodiode to verify stimulus timings, it is not possible to quantify the magnitude of such discrepancies.h p q y g p The rhythmical presentation of the target cues was varied across trials to minimise participants learning the tempo. Each trial for all conditions contained a tempo change to ensure that participants paid attention to the target cues. Six trials started with a fast, followed by a slow rhythm, and a further six trials followed the reverse order. The inter-target cue-intervals (ITI) for the fast tempo were on average 650 milliseconds (± 5%), and 870 ms (± 5%) for the slow tempo. The tempo change occurred randomly between the 5th or 7th ITI. Each condition contained 12 trials with each trial lasting 40 seconds, with an overall total of 72 trials (12 (trials) × 6 (conditions: visual social, audio social, audio-visual social, visual non-social, audio non-social, audio-visual non-social)). The presentation of both the video stimuli and the generated dot motion was controlled by Psychophysics toolbox67 in MATLAB (version 2014a; The Mathworks Inc., MA, USA).hh h The AQ was used to measure an individual’s autistic traits44. Materials and methods For the male recording the ‘ba’ sound was presented at an average of 110 Hz (minimum 108 Hz, maximum 112 Hz). Female ‘ba’ sounds were presented at an average of 211 Hz (minimum 207, maximum 215 Hz). In audio- visual social conditions, both the video and the corresponding audio recording were presented simultaneously. Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ www.nature.com/scientificreports/ Note, absolute asynchronies were used for the present analysis. For non-social conditions, absolute asynchronies were calculated by calculating the difference between the target cue event time and the participant’s corresponding response event time (see Fig. 5). Non-social target cue event times were taken from the stimulus file. For non-social audio-visual combined conditions, the target onsets for both the visual and auditory cue coincided in time. The target cue event times for visual-social condi- tions were estimated by two independent coders in Elan76. The video recordings in the visual-social conditions were presented at a rate of 30 frames per second (30fps). Coders identified the first frame of mouth opening as the target event time. For audio-social conditions of the audio data from the videos was separated and saved as a wav file. The audio data was then smoothed using a bi-directional second order Butterworth low-pass filter77. Maximum peaks were detected using an adaptive peak detector with a threshold of a valley preceding each maximum peak. Each audio target onset event was visually cross validated with a spectrogram of the raw signal70. For social stimuli, it is possible to measure synchrony using either the visual or auditory onsets for the combined audio-visual conditions. We therefore examined both synchronisation performance with visual and auditory onsets for audio-visual combined conditions (see Results). However, synchronisation performance for audio-visual conditions has previously been reported to be comparable with that of the unimodal auditory conditions20. Therefore the audio onsets, as extracted for the audio-social conditions, were used as the target cue for audio-visual condition for the primary analysis. To explore if the use of auditory vs visual onsets had a significant impact on asynchrony, we ran a separate analysis only on the social trials (model details in the fol- lowing section). g Lastly, we examined the variability of the social stimulus onsets by computing the median standard deviation across all conditions (median 0.0347 s, minimum 0.0190 s, maximum 0.220 s). Data reduction. Participants were excluded from the relevant analyses if one or more of these criteria were met: (a) being greater/less than 3 SD from the group mean (N = 1), and (b) missing data on a stimulus type for all conditions (N = 8). A linear mixed model analysis on the mean absolute asynchrony data was conducted from the remaining 42 participants. (see Table 1 for descriptives). Statistical analyses. www.nature.com/scientificreports/ The synchrony analysis adopted an information-processing approach rather than a dynamical sys- tems approach. The latter is more favoured by researchers who examine the continuous rhythmical movements. However, in the present study participants were instructed to synchronise their verbal response to match those of an external target, rather than simply to maintain a continuous rhythm. This information processing approach has been widely used by researchers who have examined synchronous performances between two or more indi- viduals; for example, to analyse finger tapping68, oscillatory arm movements69, bouncing12 and sound recording from a string quartet70. For each trial, we recorded the sound onsets for all verbal responses performed by par- ticipants. We then used a custom-made peak detection algorithm in Matlab to extract the onset times for each verbal response that occurred after the tempo change. Response data before and at time of the period change were excluded from the data analysis to reduce additional variability introduced due to a different starting tempo and adjustments made to entrain to a new tempo. The alignment of target and response onsets was achieved in a way where the closest response onset to the target onset was used to estimate asynchrony. For the first response onset the target onset would always precede the response onset. Missing responses were interpolated adopt- ing methods used in previous research71–73. The following interpolation was conducted to account for missing responses; if a participant’s response was two times as large as the target cue’s tempo (inter-onset-interval, IOI), the participant’s inter-response-interval (IRI) was split into two (divided by two). Similarly, if a participant’s response was three times as large as the target cue’s IOI, the participant’s response was divided into three equal parts to account for the missing responses. Any responses larger than three times the relative target cue’s IOI was discarded. Absolute asynchronies were calculated to indicate the magnitude of asynchrony, irrespective of a participant being ahead or behind the target stimulus74,75. Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ Figure 5. An illustration of the asynchrony calculation. Asynchrony is calculated as the difference between the event time of the respondent and the closest event time from the cue. Note, absolute asynchronies were used for the present analysis. Figure 5. An illustration of the asynchrony calculation. Asynchrony is calculated as the difference between the event time of the respondent and the closest event time from the cue. Received: 20 May 2020; Accepted: 31 March 2021 Received: 20 May 2020; Accepted: 31 March 2021 www.nature.com/scientificreports/ www.nature.com/scientificreports/ To explore the effect of learning/practice effects through the experiment, a separate model was tested, includ- ing Trial number as a predictor. The model is specified below: Mean Absolute Asynchrony ∼1 + Condition Auditory/Visual/Audio −visual Mean Absolute Asynchrony ∼1 + Condition Auditory/Visual/Audio −visual + Stimulus Type (Social/Non −social) + AQ + Gender + Trial + Condition ∗StimulusType + Stimulus Type (Social/Non −social) + AQ + Gender + Trial + Condition ∗StimulusTyp + StimulusType ∗Trial + StimulusType ∗Trial ∗AQ + 1\|Participant . + StimulusType ∗Trial + StimulusType ∗Trial ∗AQ + 1\|Participant . References k J., Fairhurst, M. T., Kotz, S. A. & Keller, P. E. Synchronizing with auditory and visual rhythms: An fMRI assessme modality differences and modality appropriateness. Neuroimage 67, 313–321 (2013). yf y g 4. 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K., Nind, L. K., Henderson, Z. & Macrae, C. N. Moving memories: Behavioral synchrony and memory for self and others J. Exp. Soc. Psychol. 46, 457–460 (2010).l p y 10. Wiltermuth, S. Synchrony and destructive obedience. Soc. Influ. 7, 78–89 (2012). p y . Wiltermuth, S. Synchrony and destructive obedience. Soc. Influ. p y 10. Wiltermuth, S. Synchrony and destructive obedience. Soc. Influ. 7, 78–89 (2012). y y fl 1. Kirschner, S. & Tomasello, M. Joint drumming: Social context facilitates synchronization in preschool children. J. Exp. Child Psychol. www.nature.com/scientificreports/ A linear mixed model implemented in jamovi v1.1.978 was defined to analyse the mean absolute asynchrony data, across all trials and after two response cycles following the tempo change in the stimulus. The model was as follows: Mean Absolute Asynchrony ∼1 + Condition Auditory/Visual/Audio −visual + Stimulus Type (Social/Non −social) + AQ + Gender + Condition ∗StimulusType + 1\|Participant . Stimulus Type (social, non-social), Condition (audio, visual or combined), Gender, AQ scores were defined as fixed effects and participants were defined as random effects. Model fit was estimated using the Restricted Maximum Likelihood (REML) method.f To check if the synchronisation performance to social combined (audio-visual) stimuli differed as a function of using auditory vs visual onsets to define the target cue, a further analysis was conducted to compare the mean asynchrony estimated by the visual and the auditory onsets respectively. This analysis was done only for trials were social stimuli were presented, since auditory and visual onsets were identical for non-social audio-visual stimuli (as programmed). The following model was estimated: Mean Asynchrony ∼1 + AQ + Condition Auditory/Visual/Audiovisual + OnsetType Auditory/Visual + Condition ∗OnsetType + 1\|Participant + OnsetType Auditory/Visual + Condition ∗OnsetType + 1\|Participant Scientific Reports \| (2021) 11:8805 \| https://doi.org/10.1038/s41598-021-88112-y www.nature.com/scientificreports/ Synchronization and leadership in string quartet performance: A case study of auditory and visual cues. Front. Psychol. 5, 645 (2014). 71. Marieke Van Der Steen, M. C., Jacoby, N., Fairhurst, M. T. & Keller, P. E. Sensorimotor synchronization with tempo-changing auditory sequences: Modeling temporal adaptation and anticipation. Brain Res. https://doi.org/10.1016/j.brainres.2015.01.053 (2015). ( ) 72. Repp, B. H. & Keller, P. E. Sensorimotor synchronization with adaptively timed sequences. Hum. Mov. Sci. 27, 423–456 (200 72. Repp, B. H. & Keller, P. E. Sensorimotor synchronization with adaptively timed sequences. Hum. Mov. 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Author contributions C d O G d d Author contributions J.J.H., B.C. and O.G. designed the experiment, J.J.H. and B.C. conducted the study, J.J.H., P.M. and B.C. analysed the data and all authors discussed the results. J.J.H., B.C. wrote the paper, and all authors reviewed the manuscript. Acknowledgements g We would like to thank Nadyne Dunkley, Makena Peart, Hristina Mihaylova, and Dr Anthony Haffey for helping with the data collection and stimuli creation, Dagmar S. Fraser for his programming support for the stimuli pres- entation, and thank all the volunteers for participation and feedback. BC was supported by UK Medical Research Council (MR/P023894/1, MR/S036423/1) during this project. https://doi.org/10.1038/s41598-021-88112-y Scientific Reports \| (2021) 11:8805 \| www.nature.com/scientificreports/ Author contributions J.J.H., B.C. and O.G. designed the experiment, J.J.H. and B.C. conducted the study, J.J.H., P.M. and B.C. analysed the data and all authors discussed the results. J.J.H., B.C. wrote the paper, and all authors reviewed the manuscript. Competing interests The authors declare no competing interests. Additional information Supplementary Information The online version contains supplementary material available at https://doi.org/ 10.1038/s41598-021-88112-y. Correspondence and requests for materials should be addressed to J.J.H. or B.C. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2021 Author contributions J.J.H., B.C. and O.G. designed the experiment, J.J.H. and B.C. conducted the study, J.J.H., P.M. and B.C. analysed the data and all authors discussed the results. J.J.H., B.C. wrote the paper, and all authors reviewed the manuscript. Additional informationh Additional information Supplementary Information The online version contains supplementary material available at https://doi.org/ 10.1038/s41598-021-88112-y. Correspondence and requests for materials should be addressed to J.J.H. or B.C. Reprints and permissions information is available at www.nature.com/reprints. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2021 https://doi.org/10.1038/s41598-021-88112-y Scientific Reports \| (2021) 11:8805 \|	31,377
https://openalex.org/W3117779584	OpenAlex	Open Science	CC-By	2,020	A Practical Guide to Incorporating Health Equity Domains in Implementation Frameworks	Eva N. Woodward	English	Spoken	17,536	30,162	A Practical Guide to Incorporating Health Equity Domains in Implementation Frameworks Eva N Woodward (  eva.woodward2@va.gov ) Central Arkansas Veterans Healthcare System Eugene J Towbin Healthcare Center Rajinder Sonia Singh US Department of Veterans Affairs Phiwinhlanhla Ndebele-Ngwenya Philander Smith College Andrea Melgar Castillo University of Arkansas for Medical Sciences South Carroll Medical Library Kelsey S. Dickson San Diego state University JoAnn E Kirchner US Department of Veterans Affairs A Practical Guide to Incorporating Health Equity Domains in Implementation Frameworks Eva N Woodward (  eva.woodward2@va.gov ) Central Arkansas Veterans Healthcare System Eugene J Towbin Healthcare Center https://orcid.org/0000-0002-7880-0054 Rajinder Sonia Singh US Department of Veterans Affairs Phiwinhlanhla Ndebele-Ngwenya Philander Smith College Andrea Melgar Castillo University of Arkansas for Medical Sciences South Carroll Medical Library Kelsey S. Dickson San Diego state University JoAnn E Kirchner US Department of Veterans Affairs Methodology Keywords: health equity, implementation, health disparities, framework, theory, implementation science, healthcare, dissemination Posted Date: June 8th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-32704/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License A Practical Guide to Incorporating Health Equity Domains in Implementation Frameworks Eva N Woodward (  eva.woodward2@va.gov ) Central Arkansas Veterans Healthcare System Eugene J Towbin Healthcare Center https://orcid.org/0000-0002-7880-0054 Rajinder Sonia Singh US Department of Veterans Affairs Phiwinhlanhla Ndebele-Ngwenya Philander Smith College Andrea Melgar Castillo University of Arkansas for Medical Sciences South Carroll Medical Library Kelsey S. Dickson San Diego state University JoAnn E Kirchner US Department of Veterans Affairs Methodology Keywords: health equity, implementation, health disparities, framework, theory, implementation science, healthcare, dissemination Posted Date: June 8th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-32704/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Methodology Posted Date: June 8th, 2020 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/36 Page 1/36 Page 1/36 Abstract Background: Due to limited systematic integration of health equity and implementation science, it is imperative to provide researchers and practitioners tools to guide implementation in settings where there is inequitable implementation of an intervention. Our prior work documented and piloted the first published adaptation of an existing implementation science framework with health equity determinants to create the Health Equity Implementation Framework. We suggested how others’ might adapt their preferred implementation science frameworks with three health equity domains: 1) cultural factors of recipients, 2) clinical encounter, or patient-provider interaction, and 3) societal influences (including but not limited to social determinants of health). This manuscript is a practical guide to utilize three health equity domains in implementation research and practice. Background: Due to limited systematic integration of health equity imperative to provide researchers and practitioners tools to guide i is inequitable implementation of an intervention. Our prior work do published adaptation of an existing implementation science frame to create the Health Equity Implementation Framework. We sugges preferred implementation science frameworks with three health eq recipients, 2) clinical encounter, or patient-provider interaction, and not limited to social determinants of health). This manuscript is a equity domains in implementation research and practice. Methods: We describe in greater depth than in our previous publication domains typical in implementation determinants frameworks and three adaptations: domains known to affect health equity. For each domain, we compiled definitions with supporting literature, defined relevant subdomains, showcased an illustrative example, and suggested sample measures, both quantitative and qualitative. Results: We describe how to incorporate the three health equity domains in one’s preferred implementation science framework, or how to use the Health Equity Implementation Framework specifically. Practical guidelines follow ten published recommendations on how to use frameworks in implementation research and practice. We describe a new case study in which the framework guided evaluation. Conclusions: Incorporating health equity domains within implementation frameworks may optimize the scientific yield and equity of implementation efforts by assessing and ideally, addressing, implementation and equity barriers simultaneously. The practical guidance and tools provided can assist implementation scientists and practitioners to concretely address inequity in implementation across populations to capture and analyze information used to assess health outcomes. Contributions to the Literature Specific definitions of implementation and three health equity domains with examples of how they have been applied in published literature and sample measures. Practical tools, including a qualitative interview guide and codebook Case study of how the Health Equity Implementation Framework guided analysis in an implementation study Background Page 2/36 Health inequities are clinically and statistically significant differences in health or health care between two or more groups.[1] Groups that experience worse health or health care might be defined by race, ethnicity, sexual orientation, gender identity, socioeconomic status, functional limitation, or other characteristics;[2] we refer to these groups as marginalized populations based on social, economic, and/or environmental disadvantage that usually accompanies health inequities. Several inequitable implementation problems may be undetected. Inequitable healthcare implementation problems exist when a healthcare innovation is delivered with significantly worse rates, access, or quality to certain groups; sometimes, the group experiencing less or worse innovations has higher prevalence of the health problem the innovation is intended to ameliorate.[3] One example in United States (U.S.) pediatric healthcare is screening and diagnosis of autism spectrum disorder to prompt access to treatment. Although there are valid and reliable autism screenings and clear criteria for diagnosis, racial and ethnic minority children who meet criteria are less likely to be diagnosed than white children.[4] Thus, effective screenings and interventions are implemented inequitably for racial and ethnic minority children. The inequity is exacerbated among children who receive an autism diagnosis, with children of color less likely to receive quality treatment.[5] Another example is intervention for maternal, newborn, and reproductive health care in low and middle income countries. Of global maternal deaths related to pregnancy, 15% occur in India.[6] Recently, an intervention with clinical effectiveness was implemented—a multifaceted social franchising model—to increase quality and coverage of maternal, newborn, and reproductive health care in one Indian state. Implementation was unsuccessful; women were not aware of the intervention and healthcare workers did not encourage women to attend services. It is possible implementation was not tailored enough to this population, and without community engagement, training, and access to resources (e.g., internet for telehealth), poor implementation and inequities persist. [7] Several inequitable implementation problems may be undetected. Inequitable healthcare implementation problems exist when a healthcare innovation is delivered with significantly worse rates, access, or quality to certain groups; sometimes, the group experiencing less or worse innovations has higher prevalence of the health problem the innovation is intended to ameliorate.[3] One example in United States (U.S.) pediatric healthcare is screening and diagnosis of autism spectrum disorder to prompt access to treatment. The Need for Practical Tools Specific to Implementation E i The need for practical implementation tools specific to equity is significant given the real-world setting of implementation research and practice, and because existing implementation tools focus little or no explicit attention on equity. Frameworks that account for equity and implementation might be too vague to be used meaningfully.[22] Interventions that work well in small-scale or controlled pilots may not perform as expected when disseminated or scaled up in real-world settings due to lack of tools necessary to inform contextual understanding, including presence, drivers, and impact of health inequities.[23] Examples of practical implementation tools include the Consolidated Framework for Implementation Research guide tool,[24] and the website to review and select and implementation framework.[25] Unfortunately, these tools have yet to be explicitly or extensively adapted, integrated with, or tested within health equity efforts. Due to limited systematic integration of health equity and implementation science, [13] we need more implementation tools with an acute awareness for equity. Our prior work documented and piloted adaptations of one existing implementation science framework with health equity determinants to create the Health Equity Implementation Framework.[11] We also included guidance on how others’ might adapt their preferred implementation science frameworks in a similar manner. Since that publication, many scientists and practitioners have requested further clarification of the Health Equity Implementation Framework domains for practical use. This manuscript provides a practical guide to utilizing the three health equity domains in a framework in settings where there is inequitable implementation. Background Although there are valid and reliable autism screenings and clear criteria for diagnosis, racial and ethnic minority children who meet criteria are less likely to be diagnosed than white children.[4] Thus, effective screenings and interventions are implemented inequitably for racial and ethnic minority children Implementation science theories, methods, strategies, and evaluation may ensure effective interventions reach the people they are designed to help. Implementation scientists and practitioners can address broad, systemic, or organizational factors in addition to patient, clinic, and hospital factors that contribute to inequities by using implementation strategies, such as learning collaboratives, audit and feedback, or changing policy.[8] However, it is probable that existing implementation strategies and the methods to select them do not reduce health inequities at this time, based on two assumptions. First, in the U.S., drawing from decades of intervention research, generic interventions—those not adapted for marginalized populations—not only often fail to reduce inequities,[9] but sometimes even exacerbate them.[10] Second, implementation science frameworks that are key to inform study design and selection of strategies to match contextual needs have only recently considered factors and processes unique to health inequities, starting with our own Health Equity Implementation Framework.[11, 12] Implementation science theories, methods, strategies, and evaluation may ensure effective interventions reach the people they are designed to help. Implementation scientists and practitioners can address broad, systemic, or organizational factors in addition to patient, clinic, and hospital factors that contribute to inequities by using implementation strategies, such as learning collaboratives, audit and feedback, or changing policy.[8] However, it is probable that existing implementation strategies and the methods to select them do not reduce health inequities at this time, based on two assumptions. Background First, in the U.S., drawing from decades of intervention research, generic interventions—those not adapted for marginalized populations—not only often fail to reduce inequities,[9] but sometimes even exacerbate them.[10] Second, implementation science frameworks that are key to inform study design and selection of strategies to match contextual needs have only recently considered factors and processes unique to health inequities, starting with our own Health Equity Implementation Framework.[11, 12] Overall, implementation science has yet to actively and systematically assess, address, and evaluate unique factors contributing to healthcare inequities, including institutional and structural problems (e.g., racism) that are economic, regulatory, social, historical, and political determinants of implementation for marginalized groups.[13] A recent workgroup on this topic recommended implementation scientists Page 3/36 Page 3/36 Page 3/36 “employ strategies and build structures that elevate equity concerns”—this can only be done when current implementation determinant frameworks purposefully incorporate health equity domains for assessment informing implementation strategies and as potential moderators for implementation intervention.[14] Outside the U.S., health equity and implementation integration predominantly involve the use of implementation frameworks in low and middle income countries or increasing delivery of services for specific populations (e.g., Indigenous communities). Regarding use of implementation frameworks in low and middle income countries, examples include a systematic review and intervention to increase the value of money in Burkina Faso and other countries,[15, 16] as well as unsuccessful implementation of an intervention to increase quality and coverage of maternal care and reproductive health in India.[7] Other examples in low and middle income countries include measurement tools normed with participants from those countries,[17] adapting interventions or delivery methods,[18] and reviewing or developing frameworks specific to those countries.[19–21] Health Equity Implementation Framework Page 4/36 Page 4/36 In the Health Equity Implementation Framework, we proposed determinants or domains believed to predict successful and equitable implementation, seen in Fig. 1.[11] Determinants are factors, levels, or components thought to partially explain why implementation would succeed or fail.[26] In the Health Equity Implementation Framework, we adapted three health equity determinant domains to the Integrated Promoting Action on Research in Implementation in Health Services (i-PARIHS) framework,[27] which also proposes a process—facilitation—by which change in each domain would occur.[26, 27] The focus of this manuscript is on three health equity determinants, rather than facilitation, as science is still emerging on how implementation processes should be tailored to promote equity. One does not need to use i-PARIHS to incorporate health equity determinants into another preferred implementation determinants framework. [insert Fig. 1 here – Health Equity Implementation Framework] Domains Typical in Implementation Determinants Frameworks. Broad domains typical in implementation determinants frameworks focus on factors at multiple levels, including the individual (e.g., personal characteristics, actors of implementation, individuals receiving an innovation), organization (e.g., clinical service, school, department, factory), community (e.g., local government, neighborhood), system (e.g., school district, hospital system) and policy (e.g., state government, broader laws).[28] These domains can be further specified, such as inner setting or outer setting within an organization.[29] Domains from i- PARIHS are the basis of the Health Equity Implementation Framework and include those typical in most implementation determinants frameworks.[28] Each domain acts to enable or constrain implementation and is defined in Table 1 with illustrative examples and sample measures. Page 5/36 Table 1 Definitions, Illustrative Examples, and Sample Measures of the Health Equity Implementation Framework Domain & Subdomains Definition Illustrative Example(s) Sample Measuresa Definitions, Illustrative Examples, and Sample M Domain & Subdomains Definition Characteristics of the Innovation[32] • Underlying knowledge sources • Clarity • Degree of fit with existing practice / values • Usability • Relative advantage • Trialability • Observable results • Evidence for the innovation[67] o Research o Clinical experiences o Patient experiences An innovation is a treatment, intervention, or practice with unique characteristics that determine how such innovations will be applied in a particular setting. Innovations fall into one of the “7 Ps”: programs, practices, principles, procedures, products, pills, or policies.[31] The innovation should be tailored to the setting’s needs and practices for successful implementation.[32] T Definitions, Illustrative Examples, and Sample Me Domain & Subdomains Definition Characteristics of the Innovation[32] • Underlying knowledge sources • Clarity • Degree of fit with existing practice / values • Usability • Relative advantage • Trialability • Observable results • Evidence for the innovation[67] o Research o Clinical experiences o Patient experiences An innovation is a treatment, intervention, or practice with unique characteristics that determine how such innovations will be applied in a particular setting. Innovations fall into one of the “7 Ps”: programs, practices, principles, procedures, products, pills, or policies.[31] The innovation should be tailored to the setting’s needs and practices for successful implementation.[32] Domain & Subdomains Sample Measuresa Sample Measuresa Quantitative: • Decision-Maker Information Needs and Preferences Survey • Electronic Health Record Nurse Satisfaction Survey[69] Qualitative: • Barriers and facilitators assessment instrument • General practitioners’ perceptions of the route of evidence- based medicine • Knowledge, attitudes, and expectations of web-assisted tobacco interventions[69] Quantitative: • Decision-Maker Information Needs and Preferences Survey • Electronic Health Record Nurse Satisfaction Survey[69] Qualitative: • Barriers and facilitators assessment instrument • General practitioners’ perceptions of the route of evidence- based medicine • Knowledge, attitudes, and expectations of web-assisted tobacco interventions[69] A study examined uptake of the Healthy Heart Kit (innovation), a risk management and patient education resource for the prevention of cardiovascular disease, in a primary care setting. They found that relative advantage (innovation was the most comprehensive tool for cardiovascular health) and observable results (evidence-based practice supports innovation) were more influential to uptake of Healthy Heart Kit than other characteristics.[68] An innovation is a treatment, intervention, or practice with unique characteristics that determine how such innovations will be applied in a particular setting. Innovations fall into one of the “7 Ps”: programs, practices, principles, procedures, products, pills, or policies.[31] The innovation should be tailored to the setting’s needs and practices for successful implementation.[32] Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 6/36 Page 6/36 Domain & Subdomains Definition Clinical Encounter (Patient- Provider Interaction) This is the nature of the interaction between patient and provider. This domain is centered on how the patient and provider choose, adapt, and coordinate the conversation to achieve their shared and personal goals concerning health related matters.[43] The interaction could be influenced by: • Predisposition features which areindividual differences that influence communication that may be objective (e.g., age) and subjective (e.g., self-concept) • Cognitive / affective influences that show how communication is related to strategy (e.g., goals), attributions (e.g., stereotypical) and trust. • Communication influences which refers to how the patient and the provider should tailor their responses to create a coherent and effective exchange.[43] Illustrative Example(s) Definition Illustrative Example(s) Sample Measuresa Clinical Encounter (Patient- Provider Interaction) This is the nature of the interaction between patient and provider. This domain is centered on how the patient and provider choose, adapt, and coordinate the conversation to achieve their shared and personal goals concerning health related matters.[43] Quantitative: In this example, the following are a few recipient characteristics: • Motivations: commitment to improve Indigenous health outcomes and desire to influence practice and policy • Values: perceived importance of group input, such as discussing data, sharing views and ideas • Knowledge: experience using Continuous Quality Improvement approaches[74] Quantitative: • Staff implementation ratings of innovation • Point of contact implementation reports assessing the innovation[75] Qualitative: • Point of contact interview[75] • The Agency of Healthcare Research and Quality guides measuring recipient perceptions about implementation[29] Illustrative Example(s) Quantitative: In studying recordings of HIV patient-provider encounters, there was less psychosocial talk in patient- provider encounters with Hispanic compared to white patients.[42] In a study on predictors and consequences of negative patient-provider interactions among a sample of African American sexual minority women, authors found racial discrimination was most frequently mentioned, and gender and sexual orientation discrimination were also related to negative patient experiences.[70] • Patient rating about encounter: Interpersonal Processes of Care Survey[42] • Predisposition features which areindividual differences that influence communication that may be objective (e.g., age) and subjective (e.g., self-concept) • Experiences of Discrimination Scale[71] Qualitative: • Cognitive / affective influences that show how communication is related to strategy (e.g., goals), attributions (e.g., stereotypical) and trust. • Communication influences which refers to how the patient and the provider should tailor their responses to create a coherent and effective exchange.[43] Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 7/36 Page 7/36 Domain & Subdomains Definition Recipients[32] • Motivations • Values and beliefs • Goals • Skills • Knowledge • Time, resources, support • Local opinion leaders • Collaboration / teamwork • Existing networks • Learning environment • Power and authority • Presence of boundaries Recipients are individuals who influence implementation processes and those who are affected by implementation outcomes, both at the individual and collective team levels. Recipients can facilitate uptake of an innovation or resist its implementation.[32] Domain & Subdomains Definition Illustrative Example(s) Sample Measuresa Recipients[32] • Motivations • Values and beliefs • Goals • Skills • Knowledge • Time, resources, support • Local opinion leaders • Collaboration / teamwork • Existing networks • Learning environment Recipients are individuals who influence implementation processes and those who are affected by implementation outcomes, both at the individual and collective team levels. Recipients can facilitate uptake of an innovation or resist its implementation.[32] An example of recipients are stakeholders in a study to identify gaps in care for Indigenous people in Australia. Illustrative Example(s) Sample Measuresa Sample Measuresa A providers’ objectives and beliefs about a patient affects how they behave during the patient-provider interaction.[76] Providers, especially in busy healthcare settings, may be vulnerable to subconscious bias and stereotypes.[77] Physicians who consider themselves “liberal” spent more time giving more information to patients th those who consider themselves “conservative [43] Providers may engage in more detailed conversatio about the health status of educated patients, yet provide basic explanation for less educated patients [43] During a post-angiogram encounter, physicians perceived patients of lowe socioeconomic status as having more negative personality characteristics that include lack of self- control and more negative behavioral tendencies.[41 Domain & Subdomains Definition Recipients: Providers and staff Culturally relevant factors include:[38] • Demographics (e.g., neighborhood immigrant status) • Unconscious / implicit bias • Knowledge and attitudes • Skillsets In a healthcare setting, providers and staff are the people who administer the innovation. A providers’ objectives and beliefs about a patient affects how they behave during the patient-provider interaction.[76] Providers, especially in busy healthcare settings, may be vulnerable to subconscious bias and stereotypes.[77] Sample Measuresa An example of recipients are stakeholders in a study to identify gaps in care for Indigenous people in Australia. In this example, the following are a few recipient characteristics: • Motivations: commitment to improve Indigenous health outcomes and desire to influence practice and policy • Values: perceived importance of group input, such as discussing data, sharing views and ideas • Knowledge: experience using Continuous Quality Improvement approaches[74] Quantitative: • Staff implementation ratings of innovation • Point of contact implementation reports assessing the innovation[75] Qualitative: • Point of contact interview[75] • The Agency of Healthcare Research and Quality guides measuring recipient perceptions about implementation[29] An example of recipients are stakeholders in a study to identify gaps in care for Indigenous people in Australia. In this example, the following are a few recipient characteristics: • Motivations: commitment to improve Indigenous health outcomes and desire to influence practice and policy • Values: perceived importance of group input, such as discussing data, sharing views and ideas • Knowledge: experience using Continuous Quality Improvement approaches[74] Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 8/36 Page 8/36 Domain & Subdomains Definition Illustrative Example(s) Recipients: Providers and staff Culturally relevant factors include:[38] • Demographics (e.g., neighborhood immigrant status) • Unconscious / implicit bias • Knowledge and attitudes • Skillsets In a healthcare setting, providers and staff are the people who administer the innovation. Sample Measuresa Quantitative:[37] • Telephone survey of a random sample of residents • Medical Mistrust Index[90] • Measures of underutilization of health services • Health literacy question[91] • Health numeracy question[92] • Appropriated Racial Oppression Scale[93] Qualitative: • Interview about expectations for treatment or the patient-provider- interaction[42, 94] • Interviews about experience seeking care[95] Domain & Subdomains Sample Measuresa Sample Measuresa In a healthcare setting, providers and staff are the people who administer the innovation. Physicians who consider themselves “liberal” spent more time giving more information to patients than those who consider themselves “conservative”. [43] Physicians who consider themselves “liberal” spent more time giving more information to patients than those who consider themselves “conservative”. [43] A providers’ objectives and beliefs about a patient affects how they behave during the patient-provider interaction.[76] Providers may engage in more detailed conversations about the health status of educated patients, yet provide basic explanations for less educated patients. [43] Providers, especially in busy healthcare settings, may be vulnerable to subconscious bias and stereotypes.[77] During a post-angiogram encounter, physicians perceived patients of lower socioeconomic status as having more negative personality characteristics that include lack of self- control and more negative behavioral tendencies.[41] • One-on-one interviews[83] Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 9/36 Page 9/36 Domain & Subdomains Recipients: Patients Culturally relevant factors include: [84–86, 38, 37, 36] • Medical mistrust • Health literacy and numeracy • Demographics (e.g., neighborhood, immigrant status) • Socioeconomic status, including household income, net wealth, health insurance status, education level • Expectations about therapeutic relationships • Biology/genetics Domain & Subdomains Recipients: Patients Culturally relevant factors include: [84–86, 38, 37, 36] • Medical mistrust • Health literacy and numeracy • Demographics (e.g., neighborhood, immigrant status) • Socioeconomic status, including household income, net wealth, health insurance status, education level • Expectations about therapeutic relationships • Biology/genetics Illustrative Example(s) Illustrative Example(s) Quantitative:[37] Asian American patients in Hawaii participated less in their medical visits than mainland Americans.[87] In a healthcare setting, patients are the people (individuals, families, caregivers) who will actually receive the innovation. Culturally relevant factors are associated with health and health care disparities and can include demographic factors, beliefs, information, and biological or genetic conditions related to equitable implementation. In a healthcare setting, patients are the people (individuals, families, caregivers) who will actually receive the innovation. Culturally relevant factors are associated with health and health care disparities and can include demographic factors, beliefs, information, and biological or genetic conditions related to equitable implementation. Patients with more formal educations are more expressive and tend to want to play a role in the decision- making process than less educated patients.[43] Many patients are unsure about their role in the encounter and the appropriateness of their participation.[88] We have limited ability to predict treatment success based on whether research was conducted with patients of similar clinical phenotype.[89] Page 10/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 10/36 Page 10/36 Domain & Subdomains Definition Illustrative Example(s) Inner context (local)[27] • Formal and informal leadership support • Culture • Previous experience of innovation or change • Change mechanisms for embedding innovation • Evaluation and feedback processes The immediate local setting of implementation. Examples include: • Ward • Unit • Clinic • Hospital department Among 303 providers working in 49 publicly funded health programs for youths, providers’ perception of certain leadership styles were associated with stronger provider willingness to adopt evidence-based treatments. Quantitative:[37] [96] Pisando Fuerte is a fall prevention program linguistically and culturally tailored for Latino individuals at risk for falls. It is adapted from “Stepping On,” an evidence-based fall prevention program. Fidelity to Pisando Fuerte was subpar; when comparing fidelity between two sites, fidelity was lower in the site that did not give additional time to implement the program (poor leadership support) and had no experience in organizing programs like Pisando Fuerte (no previous experience of innovation). [97] Sample Measuresa Sample Measuresa tional n which am is Hospitals’ adoption of the Culturally and Linguistically Appropriate Services standards focused on retaining translators and adapting culturally and linguistically appropriate materials. However, this adoption did not often include engagement in broader organizational change.[104] Researchers studied a disparity-reduction program in Israel across 26 clinics and 109 clinical teams. After three years, they found different inner context configurations of factors predicting disparity reduction. One example of a successful configuration were: clinics with a large disparity gap to minimize, high clinic density, high perceived team effectiveness, focused efforts on tailoring services to their enrollees patients.[105] Quantitative: • Measures of organizational readiness for change[106] • Cultural Competency Assessment Tool for Hospitals[104] Qualitative:[107] • Key informant interviews assessing knowledge/action of policies about equity • Key informant interviews assessing beliefs organization holds about marginalized people • Stakeholder meetings about importance of equitable care • Public forums & listening sessions[108] • Focus groups The organizational atmosphere in which the unit or team is embedded. Hospitals’ adoption of the Culturally and Linguistically Appropriate Services standards focused on retaining translators and adapting culturally and linguistically appropriate materials. However, this adoption did not often include engagement in broader organizational change.[104] Researchers studied a disparity-reduction program in Israel across 26 clinics and 109 clinical teams. After three years, they found different inner context configurations of factors predicting disparity reduction. One example of a successful configuration were: clinics with a large disparity gap to minimize, high clinic density, high perceived team effectiveness, focused efforts on tailoring services to their enrollees patients.[105] Page 12/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Illustrative Example(s) Pisando Fuerte is a fall prevention program linguistically and culturally tailored for Latino individuals at risk for falls. It is adapted from “Stepping On,” an evidence-based fall prevention program. Fidelity to Pisando Fuerte was subpar; when comparing fidelity between two sites, fidelity was lower in the site that did not give additional time to implement the program (poor leadership support) and had no experience in organizing programs like Pisando Fuerte (no previous experience of innovation). [97] • Key informant interviews about inclusivity Page 11/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 11/36 Page 11/36 Domain & Subdomains Definition Illustrative Example(s) Inner context (organizational)[27] • Organizational priorities • Senior leadership and management support • Culture • Structure and systems • History of innovation and change • Absorptive capacity • Learning networks The organizational atmosphere in which the unit or team is embedded. Hospitals’ adoption of the Culturally and Linguistically Appropriate Services standards focused on retaining translators and adapting culturally and linguistically appropriate materials. However, this adoption did not often include engagement in broader organizational change.[104] Researchers studied a disparity-reduction program in Israel across 26 clinics and 109 clinical teams. After three years, they found different inner context configurations of factors predicting disparity reduction. One example of a successful configuration were: clinics with a large disparity gap to minimize, high clinic density, high perceived team effectiveness, focused efforts on tailoring services to their enrollees patients.[105] The organizational atmosphere in which the unit or team is embedded. Sample Measuresa Sample Measuresa Qualitative: • Archival analysis, reading and documenting policies, program manuals, or procedural protocols[109, 110] • Interviews with leadership[105] Quantitative: • 15 core measures of health care quality[111] • Population surveys • Social network analysis of relationships between relevant leadership and/or teams [105] • Existing reports hospital-wide scores on assessments of care and equity, e.g., National Quality Forum or Healthcare Equality Index[112] Sample Measuresa Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 12/36 Page 12/36 Domain & Subdomains Definition Outer context (healthcare system)[27] • Policy drivers and priorities • Incentives and mandates • Regulatory frameworks or external accreditation systems • Inter- organizational networks and relationships This is the broader context defined in terms of resources, culture, leadership and orientation to evaluation and learning. There is an increasing amount of research that shows that inequities in obtaining preventative care among minorities compared with whites are due to “organizational characteristics, including location, resources, and complexity of a clinic or practice.”[38] Illustrative Example(s) Illustrative Example(s) Sample Measuresa Illustrative Example(s) Sample Measuresa Sample Measuresa In piloting the Health Equity Implementation Framework, societal influences affected receipt of antiviral Hepatitis C Virus medicine for Black patients in the U.S. Veterans Health Administration. One economic facilitator was free Hepatitis C Virus treatment for patients instituted by policymakers. One sociopolitical barrier to implementation was stigma about being diagnosed with Hepatitis C Virus.[11] Qualitative: This is the broader context defined in terms of resources, culture, leadership and orientation to evaluation and learning. Researchers examined predisposing, enabling, and need factors as predictors of changes in health care utilization, and found that patients’ experiences differed by group within the healthcare system, and impacted their beliefs and attitudes about receiving healthcare, ultimately affecting the extent to which healthcare services were utilized.[70] Quantitative: Page 13/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 13/36 Page 13/36 Domain & Subdomains Definition Societal Influences[35, 44– 46] • Economies • Physical structures • Sociopolitical forces • Up-, mid-, or downstream social determinants of health[35] Forces outside the healthcare system th influence all other domains and subdomains determining success implementation. May include but be broade than social determinants of heal May focus on presen of stigma and discrimination such a racism, classism, or transphobia (as examples) and the institutionalization o such discrimination i every subdomain of implementation.b,c Illustrative Example(s) Sample Measuresa In piloting the Health Equity Implementation Framework, societal influences affected receipt of antiviral Hepatitis C Virus medicine for Black patients in the U.S. Veterans Health Administration. One economic facilitator was free Hepatitis C Virus treatment for patients instituted by policymakers. One sociopolitical barrier to implementation was stigma about being diagnosed with Hepatitis C Virus.[11] See below Sample Measuresa d e Quantitative: • Insurance claims data • Gross Domestic Product[114] • Gross National Product[115] • Minimum Wage[116] • Population & total employment[117] • Annual average wage level of the primary, secondary, and tertiary industries[118] • Tax revenue as percentage of total revenue[119] • Interest rate on saving deposits and inflation rate[120] Qualitative:[121] • Key informant interviews about goods and services exchanged[122] • Analysis of comparative economic structure[121] cture of the e, or country o the wealth urces of nd what is ed for re delivery urance). This vided into esources (i.e., anagement) human s (i.e., land, oods, financial s, and gy).[52] In a study assessing longitudinal effects of health insurance and poverty, researchers reported low-income, middle-aged adults in the U.S. with no insurance, unstable coverage, or changes in insurance have higher out- of-pocket expenditures and financial burdens than public insurance holders, as well as increased financial burden.[113] In a case study, the presence of chronic kidney disease indicators in the pay-for- performance system in primary care created an incentive for improvement. [27] Quantitative: • Insurance claims data • Gross Domestic Product[114] • Gross National Product[115] • Minimum Wage[116] • Population & total employment[117] • Annual average wage level of the primary, secondary, and tertiary industries[118] • Tax revenue as percentage of total revenue[119] • Interest rate on saving deposits and inflation rate[120] Qualitative:[121] • Key informant interviews about goods and services exchanged[122] • Analysis of comparative economic structure[121] In a study assessing longitudinal effects of health insurance and poverty, researchers reported low-income, middle-aged adults in the U.S. with no insurance, unstable coverage, or changes in insurance have higher out- of-pocket expenditures and financial burdens than public insurance holders, as well as increased financial burden.[113] In a case study, the presence of chronic kidney disease indicators in the pay-for- performance system in primary care created an incentive for improvement. [27] Page 15/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. See below In piloting the Health Equity Implementation Framework, societal influences affected receipt of antiviral Hepatitis C Virus medicine for Black patients in the U.S. Veterans Health Administration. One economic facilitator was free Hepatitis C Virus treatment for patients instituted by policymakers. One sociopolitical barrier to implementation was stigma about being diagnosed with Hepatitis C Virus.[11] Forces outside the healthcare system that influence all other domains and subdomains determining success of implementation. May include but be broader than social determinants of health. May focus on presence of stigma and discrimination such as racism, classism, or transphobia (as examples) and the institutionalization of such discrimination in every subdomain of implementation.b,c Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 14/36 Page 14/36 Domain & Subdomains Definition Economies[49] • Traditional • Command • Market • Mixed The structure of the city, state, or country related to the wealth and resources of people and what is exchanged for healthcare delivery (e.g., insurance). This can be divided into human resources (i.e., labor, management) and nonhuman resources (i.e., land, capital goods, financia resources, and technology).[52] Domain & Subdomains Definition Economies[49] • Traditional • Command • Market • Mixed The structure of the city, state, or country related to the wealth and resources of people and what is exchanged for healthcare delivery (e.g., insurance). This can be divided into human resources (i.e., labor, management) and nonhuman resources (i.e., land, capital goods, financial resources, and technology).[52] Illustrative Example(s) Illustrative Example(s) The physical environment, structure, location of services and recipients. Also known as the built environment as it relates to equitable implementation.[52] Sample Measuresa Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 15/36 Page 15/36 Domain & Subdomains Definition Physical structures • Location • Availability of public transportation • Actual environment of the point-to-care • Language spoken and/or signage • Available structures in one’s neighborhood to use innovation • Grocery stores • Health care facilities • Local businesses • Physical infrastructure The physical environment, structur location of services and recipients. Also known as the built environment as it relates to equitable implementation.[52] Domain & Subdomains Definition Illustrative Exam Physical structures • Location • Availability of public transportation • Actual environment of the point-to-care • Language spoken and/or signage • Available structures in one’s neighborhood to use innovation • Grocery stores • Health care facilities • Local businesses • Physical infrastructure The physical environment, structure, location of services and recipients. Also known as the built environment as it relates to equitable implementation.[52] In a study comp and white peop one U.S. city, au reported that lo residence may inequities. For e the racially inte income neighbo inequities in hy diabetes, obesit women, and us services signific decreased betw and white Ame In a qualitative transgender ind experiences in r addiction treatm researchers obs residential facil split the milieu based on the ge may be stigmat who identify as or gender non-c [124] Domain & Subdomains Physical structures • Location • Availability of public transportation • Actual environment of the point-to-care • Language spoken and/or signage • Available structures in one’s neighborhood to use innovation • Grocery stores • Health care facilities • Local businesses • Physical infrastructure Illustrative Example(s) Illustrative Example(s) Definition Sample Measuresa Sample Measuresa Sample Measuresa In a study comparing Black and white people living in one U.S. city, authors reported that location of residence may affect health inequities. For example, in the racially integrated, low- income neighborhood, inequities in hypertension, diabetes, obesity among women, and use of health services significantly decreased between Black and white Americans.[123] In a qualitative study of transgender individuals’ experiences in residential addiction treatment, researchers observed that residential facilities that split the milieu and housing based on the gender binary may be stigmatizing people who identify as transgender or gender non-conforming. [124] Quantitative: • Indices of Segregation[125]b • Public data such as: Hospitals per capita, Public transportation trips per capita, Car ownership, Revenue dedicated to parks & recreation, transportation, other infrastructure needs, Grocery stores per capita • Center on Budget and Policy Priorities data • State Departments of Finance and Administration[52] Qualitative: • Windshield & walking surveys include assessing infrastructure; surveyors are on foot and take note of the neighborhood related to physical or built environment.[126] Quantitative: • Indices of Segregation[125]b • Public data such as: Hospitals per capita, Public transportation trips per capita, Car ownership, Revenue dedicated to parks & recreation, transportation, other infrastructure needs, Grocery stores per capita • Center on Budget and Policy Priorities data • State Departments of Finance and Administration[52] Qualitative: • Windshield & walking surveys include assessing infrastructure; surveyors are on foot and take note of the The physical environment, structure, location of services and recipients. Also known as the built environment as it relates to equitable implementation.[52] In a study comparing Black and white people living in one U.S. city, authors reported that location of residence may affect health inequities. For example, in the racially integrated, low- income neighborhood, inequities in hypertension, diabetes, obesity among women, and use of health services significantly decreased between Black and white Americans.[123] In a qualitative study of transgender individuals’ experiences in residential addiction treatment, researchers observed that residential facilities that split the milieu and housing based on the gender binary may be stigmatizing people who identify as transgender or gender non-conforming. [124] Qualitative: • Windshield & walking surveys include assessing infrastructure; surveyors are on foot and take note of the neighborhood related to physical or built environment.[126] Page 16/36 Measures or data collection methods are examples from literature; for a repository of ntation science measures, see the Society for Implementation Research Collaboration’s nt Review Project.[129] b. Policies and procedures, formal or informal, in national and local governments that systemically inhibit or promote equitable health. Sample Measuresa For a repository of measures specific to racism, see Appendix B of Science & Tools for the Public Health Professional.[130] c. Implementation scientists should xisting measurement tools specific to health disparities in your area of interest or study to tegrate health equity into implementation. Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Page 16/36 Page 16/36 Domain & Subdomains Definition Sociopolitical Forces[44, 46, 55] • Policy climate • Political support • Laws • Local culture • Social movements or structures such as racism, classism, heterosexism, transphobiac Policies and procedures, formal or informal, in national and local governments that systemically inhibit or promote equitable health. Notes. a. Measures or data collection methods are implementation science measures, see the Society Instrument Review Project.[129] b. For a repository Racism: Science & Tools for the Public Health Pro review existing measurement tools specific to hea further integrate health equity into implementation Innovation. Innovation refers to the treatment, interv Illustrative Example(s) Sample Measuresa There are processes by which barriers in implementation domains are solved or overcome, and strengths are harnessed to promote use of an innovation in routine practice.[29] In i- PARIHS, facilitation is the “active ingredient” or process.[32] Facilitation involves implementation strategies that result in implementation coming to fruition.[33, 34] Domains Known to Affect Health Equity. Three health equity domains have clear, strong associations with inequities in health status, access to, quality of, or outcomes of health care,[35] or there is enough evidence to suggest these domains should be considered.[e.g., 36] The Health Equity Implementation Framework incorporates these domains known to affect health equity: 1) cultural factors, such as medical mistrust, demographics, or biases of recipients, who include patients, providers, and others;[37– 40] 2) clinical encounter or patient-provider interaction;[41–43] and 3) societal influences including physical structures, economies, and social and political forces.[44–46] See Table 1 for illustrative examples and sample measures. Cultural factors of recipients. Recipients in the implementation process are individuals who will consume a particular innovation (e.g., patients, providers).[27] Recipients have unique characteristics, as single individuals and as a group, that influence whether an innovation will be adapted and adopted successfully. Implementers should tailor implementation strategies so they are culturally aligned with beliefs, behaviors, preferences, and needs of all recipients—providers, staff, and patients. Some examples of recipient cultural factors are implicit bias, socioeconomic status, race and/or ethnicity, immigrant acculturation, language, health literacy, health beliefs, or trust in the clinical staff or patient group.[39, 40] Although cultural factors will vary by group and local context, it is crucial recipient cultural factors are considered, particularly when patients belong to a group experiencing a healthcare inequity. Clinical encounter (patient-provider interaction). This domain recognizes the transaction that occurs between patients and provider in health care, where decisions concerning diagnoses and treatment are made, and providers administer care.[47] It is important the recipients tailor and adapt the conversation accordingly to achieve their individual and shared goals.[43, 48] Factors from both the patient and provider include individual differences in, for example, age and self-concept, pre-existing stereotypes, or lack of trust that could hinder the interaction.[43] There could be subconscious, implicit, or explicit bias from either recipient due to the others’ characteristics, such as race, weight, or perceived sexual orientation. Due to the myriad factors intersecting during the clinical encounter, it is crucial to assess and address it, especially with regards to health inequities. Societal influence. Sample Measuresa Quantitative: • The State-Level Racism Index[127]b Qualitative: • INCLENS equity lens: examines whether clinical guidelines address health needs and inequities experienced by marginalized groups[128] • Interview questions with recipients about laws, policies, or social movements relevant to the innovation • Archival analysis[109, 110] f Policies and procedures, formal or informal, in national and local governments that systemically inhibit or promote equitable health. In a U.S. study on adoption of behavioral health evidence based treatment by states, the following were some factors that played a role: state characteristics, state fiscal supports to promote innovation adoption, and state policy supports to promote evidence based treatment adoption.[55] • Archival analysis[109, 110] Notes. a. Measures or data collection methods are examples from literature; for a repository of implementation science measures, see the Society for Implementation Research Collaboration’s Instrument Review Project.[129] b. For a repository of measures specific to racism, see Appendix B of Racism: Science & Tools for the Public Health Professional.[130] c. Implementation scientists should review existing measurement tools specific to health disparities in your area of interest or study to further integrate health equity into implementation. Innovation. Innovation refers to the treatment, intervention, or practice or new “thing” to be implemented, adopted by providers and staff, and delivered to patients.[30] The innovation may be a program, practice, principle, procedure, product, pill, or policy.[31] Innovation. Innovation refers to the treatment, intervention, or practice or new “thing” to be implemented, adopted by providers and staff, and delivered to patients.[30] The innovation may be a program, practice, principle, procedure, product, pill, or policy.[31] Recipients. Recipients are the individuals who influence implementation processes and those who are affected by implementation outcomes, both at the individual and collective team levels.[27] In health care, recipients are often many stakeholders typically grouped into providers and other staff, and patients and caregivers. Context. Context includes different micro, meso, or macro levels that correspond to inner and outer contexts often specified in determinant frameworks.[27] Context can include factors such as resources, culture, leadership, and orientation to evaluation and learning. In this framework, the micro level includes local inner context (e.g., specific ward or clinic), whereas the meso includes the organization (e.g., Page 17/36 hospital or medical center). The macro level of outer context includes the wider healthcare system and effect this has on the other domains (e.g., United Kingdom National Health Service).[28] Facilitation or Process. Sample Measuresa Societal influence includes three subdomains: 1) economies, 2) physical structures, and 3) sociopolitical forces. This domain is similar to social determinants of health, yet incorporates more upstream determinants (e.g., governance) that have been investigated less relative to mid- or downstream determinants (e.g., neighborhoods).[35] Societal influences also include historical or current discrimination against marginalized groups, such as racism, classism, or transphobia, that may be Page 18/36 Page 18/36 formally or informally institutionalized within any implementation subdomain. These factors usually occur in broadest levels of the environment (e.g., province, nation), thus affecting downstream the healthcare system, clinics, and recipients. Many societal influences are interrelated, such as a policy affecting a physical structure. It is not as important at this point to distinguish whether a factor is exclusively an economy, physical structure, social norm, or all three; rather, it is important these societal influences are considered and addressed to ensure strategies address these key drivers of societal inequities. For example, Latinx patients in community mental health care (recipients, some immigrants, some Spanish speaking) interface with a service system (purveyor of mental health services, potentially an economic factor). There may be limited purveyors of services in certain geographic regions (physical structure), with higher levels of perceived racism among some providers (social discrimination interactin with recipient factor), and limited purveyors that offer culturally or linguistically aligned care to match needs of monolingual patients who have immigrated to the U.S. (physical structure). Societal influences may not all be assessed in one study or initiative using established measures, due to feasibility constraints, but they should be documented in formative evaluations / initial diagnostic assessments of the implementation problem. Economies. There are four typical structures of economies including a traditional economy (i.e., mostly agricultural), market economy (i.e., firms and private interests control capital), command economy (i.e., government controls capital), and a mixed economy (combination of command and market).[49] It is helpful to consider how economic structure affects access to resources for implementation. Market forces can be used to change demand for products deemed healthy or unhealthy, therefore driving policy implementation. Examples of market forces include taxes on tobacco, unhealthy food, and soft drinks, or food subsidy programs for women with low incomes.[44] As another example, one study found poverty was concentrated in the U.S. Sample Measuresa central Appalachia region where mountain top mining was the main income source.[50] These residents had increased exposure to environmental hazards combined with limited resources to address negative health outcomes as a result of those hazards. Implementation in this region would need to account for resources available to residents, their unique health concerns, and healthcare in their local context. Physical structures. Equity can be affected by how physical spaces, or “built environments,” are arranged and how transition between those spaces occurs for health care.[44] One example in healthcare settings is type and quality of language translations of information displayed (e.g., flyers, waiting rooms)— whether it matches the language of patients served.[51] The location of the healthcare setting in a town or city is important in relation to location of patients[51, 52] e.g., is it difficult for patients to get to the point of care? An example even broader includes an air pollution intervention implemented in the U.S. in which low emission zones improved air quality and had positive effects on mortality rates for all residents, but a greater benefit for the wealthiest, widening mortality inequities.[53] Another example is the implementation of one U.S. state’s naloxone standing order in which pharmacies could distribute naloxone without a prescription.[54] Results demonstrated 61.7% of retail pharmacies had naloxone available without a prescription. However, naloxone availability was lower in neighborhoods with higher percentages of residents with public health insurance—partially a physical structure problem. This finding Page 19/36 was particularly problematic due to an increased cost of naloxone for people on Medicaid (public health insurance) as a result of the statewide mandate. These examples suggest health inequity can be maintained or widened through implementation efforts, such that the most affluent experience a better outcome than others. was particularly problematic due to an increased cost of naloxone for people on Medicaid (public health insurance) as a result of the statewide mandate. These examples suggest health inequity can be maintained or widened through implementation efforts, such that the most affluent experience a better outcome than others. Sociopolitical forces. Sample Measuresa Other societal influences are social norms or political forces, which can include but are not limited to political support, laws, and social structures such as racism, misogyny, classism, or heterosexism.[46, 55] Public health policies (e.g., fiscal, regulation, education, preventative treatment, and screening) demonstrate positive and negative effects on health inequalities that occur across health domains (e.g., tobacco, food and nutrition, reproductive health services).[44] As another example, a study examined U.S. state legislators’ behavioral health research-seeking practices and dissemination preferences and found significant variation between Democrats and Republicans, suggesting dissemination materials be tailored to different social norms for different groups.[56] Another example is how ableism (discrimination and stigma against people with mental or physical disabilities) acts as a barrier to implementation.[46] Sociopolitical forces. Other societal influences are social norms or political are not limited to political support, laws, and social structures such as racis heterosexism.[46, 55] Public health policies (e.g., fiscal, regulation, educatio screening) demonstrate positive and negative effects on health inequalitie domains (e.g., tobacco, food and nutrition, reproductive health services).[44 examined U.S. state legislators’ behavioral health research-seeking practice preferences and found significant variation between Democrats and Repub dissemination materials be tailored to different social norms for different g how ableism (discrimination and stigma against people with mental or phy barrier to implementation.[46] How to Use the Three Health Equity Domains as Determinants of Equitable Implementation Three health equity domains can be applied to one’s preferred implementation determinants framework. Implementation determinant frameworks need well-grounded constructs that are clearly defined, labeled, and described, with suggested validated measures[57]—we provide this in Table 1 for the Health Equity Implementation Framework. We will describe how to use the three health equity domains according to ten recommendations on using implementation frameworks in research and practice.[58] Practical tools are presented in additional files. Refer to Table 2 for an overview of all ten recommendations and definitions. We expand in text only on steps relevant for using the three health equity domains. Page 20/36 Table 2 Table 2 Ten Recommendations on Using Implementation Frameworks Step Definition 1. Select a suitable framework. Choose a framework relevant and for your implementation question and population. 2. Establish and maintain community stakeholder engagement and partnerships. Engage community members to develop/select, adapt, evaluate, and implement an intervention.[11] 3. Define issue and develop research or evaluation questions and hypotheses. Specify the problem for the specific population and intervention you plan to implement. 4. Develop an implementation mechanistic process model or logic model. Use your framework to identify and map out the process and different domains through which you believe the intervention will become implemented. 5. Select research and evaluation methods. Identify methods and evaluation tools that are specific to the intervention and population you plan to engage for implementation. There exist for populations experiencing healthcare inequities all the usual research challenges for measuring improvement in health or healthcare, and low prevalence of some issues (e.g., suicide) or low numbers of a population (e.g., gender nonconforming people) make measurement harder. Andreson et al (2004) suggested sampling and recruitment techniques for populations that experience health inequities and are hard to reach with traditional techniques (e.g., periodic population-based surveys, Integrated Disease Surveillance and Response).[90] It may also be helpful to incorporate mixed methods assessment of multiple levels to “illuminate plausible causal mechanisms.”[63] 6. Determine implementation factors/determinants. Carry out a formative evaluation and assess barriers and facilitators for relevant domains. 7. Select and tailor, or develop, implementation strategy(s). Sort or categorize barriers by framework domains then select implementation strategies to address each barrier by framework domain. 8. Specify implementation outcomes and evaluate implementation. It can be helpful to select an evaluation framework to assess outcomes, as well as selection of outcomes relevant to the implementation. 9. How to Use the Three Health Equity Domains as Determinants of Equitable Implementation Write a scientific and/or program report using the Implementation Framework Utilization Checklist as a final documentation of an implementation effort. Step Step 1. Select a suitable framework. The Health Equity Implementation Framework (or three health equity domains adapted to another implementation framework) is population and country agnostic; domains can be adapted to any setting where inequitable implementation occurs. The framework proposes determinants of inequitable implementation and a process (facilitation) by which to address determinants, but it is not an evaluation framework and not suitable for selecting evaluation outcomes. Steps 3–5. Define issue and develop research or evaluation questions and hypotheses; Develop an implementation mechanistic process model or logic model; Select research and evaluation methods. For quantitative research questions, between-site or between-group comparisons may answer questions about equitable delivery of care.[59] Potential questions of interest to equitable implementation are: Do specific people have more access to the innovation who did not before (i.e., reach)? Are systemic changes (e.g., policy changes, increased training for providers) occurring (i.e., adoption)? Are inequities between low- and well-resourced sites or patients decreasing or non-significant (effectiveness or implementation fidelity)?[60] For step 4 (mechanisms), using the three health equity domains (added to another implementation framework, as done in the Health Equity Implementation Framework), one may develop theoretically driven hypotheses about which domains or mechanisms must change to lead to improved equity and implementation success.[61] When working on an inequitable implementation problem, this will ensure some mechanisms related to equity and implementation are investigated. For example, there may be inequity in the research-practice gap related to outer context (e.g., this intervention is only implemented in well-funded institutions that serve mostly affluent people). Clearly identify patient recipients intended to receive the innovation and if there is a subset within this population who experience inequitable access to, quality of, or outcomes of healthcare. Research and evaluation methods should match existing evidence of the intervention and implementation, but also the evidence related to the inequity or marginalized population. To select an implementation research design, consider the decision tree from Chinman and colleagues.[62] Consult Table 1 for sample measures as possible measures or methods. Step 6. Determine implementation factors/determinants. This recommendation is most closely aligned with use of Health Equity Implementation Framework to date.[11] If one is using a different implementation framework for a problem in which inequity occurs, adapt the framework to include the three health equity domains described above. How to Use the Three Health Equity Domains as Determinants of Equitable Implementation Use a framework(s) at As implementation progresses, the implementation plan needs to be altered as determinants change Table Ten Recommendations on Using Step Definition 1. Select a suitable framework. Choose a framework releva population. 2. Establish and maintain community stakeholder engagement and partnerships. Engage community membe implement an intervention. 3. Define issue and develop research or evaluation questions and hypotheses. Specify the problem for the implement. 4. Develop an implementation mechanistic process model or logic model. Use your framework to iden domains through which yo implemented. 5. Select research and evaluation methods. Identify methods and evalu and population you plan to populations experiencing h challenges for measuring im prevalence of some issues (e.g., gender nonconformin et al (2004) suggested sam that experience health ineq techniques (e.g., periodic po Surveillance and Response methods assessment of m mechanisms.”[63] 6. Determine implementation factors/determinants. Carry out a formative evalu relevant domains. 7. Select and tailor, or develop, implementation strategy(s). Sort or categorize barriers b implementation strategies t 8. Specify implementation outcomes and evaluate implementation. It can be helpful to select a well as selection of outcom 9. Use a framework(s) at micro level to conduct and tailor implementation. As implementation progres as determinants change. Use your framework to identify and map out the process and different domains through which you believe the intervention will become implemented. Identify methods and evaluation tools that are specific to the intervention and population you plan to engage for implementation. There exist for populations experiencing healthcare inequities all the usual research challenges for measuring improvement in health or healthcare, and low prevalence of some issues (e.g., suicide) or low numbers of a population (e.g., gender nonconforming people) make measurement harder. Andreson et al (2004) suggested sampling and recruitment techniques for populations that experience health inequities and are hard to reach with traditional techniques (e.g., periodic population-based surveys, Integrated Disease Surveillance and Response).[90] It may also be helpful to incorporate mixed methods assessment of multiple levels to “illuminate plausible causal mechanisms.”[63] Carry out a formative evaluation and assess barriers and facilitators for relevant domains. Page 21/36 Step Definition 10. Write the proposal and report. Write a scientific and/or program report using the Implementation Framework Utilization Checklist as a final documentation of an implementation effort. Step Definition 10. Write the proposal and report. Box 1. Application of the Health Equity Implementation Framework to Guide an Equity-Focused Implementation Project: A Case Study Box 1. Application of the Health Equity Implementation Framework to Guide an Equity-Focused Implementation Project: A Case Study The Health Equity Implementation Framework has been applied to guide evaluation of an ongoing implementation project aiming to reduce inequities in the provision and receipt of publicly funded services for individuals with developmental disabilities. In 2016, the State of California Department of Developmental Services made funds available to address significant inequities in service expenditures for Latinx clients. In response, local researchers initiated a partnership with the San Diego Regional Center, the primary local purveyor of publicly-funded developmental disability services, to identify inequity reduction targets and develop and implement an inequity reduction model. A mixed-methods needs assessment was conducted to inform model development and activities. Quantitative data was gathered from secondary data extraction and analyses of the purveyor’s service utilization data for the previous year. Focus groups with purveyor case managers were held to identify key determinants and sources of inequities from their perspectives. The Health Equity Implementation Framework was applied to guide identification of implementation determinants and selection of data coding and analyses for this needs assessment (Steps 5 and 6 described above). Specifically, the framework informed the development of the qualitative codebook, including identification of coding domains and definitions that were then iteratively refined and tailored for this project. The framework also guided subsequent mixed-methods data merging and integration, including use of qualitative themes to complement and expand quantitative findings. Step 8. Specify implementation outcomes and evaluate implementation. Evaluation frameworks can be helpful in selecting implementation outcomes,[26] and it is essential to also select outcomes to evaluate equity in access to, quality of, or outcomes of implementation.[38, 59] The three health equity domains (in addition to typical implementation domains) can be helpful in designing qualitative codebooks or templates for analysis or re-evaluating quantitative measures first collected during formative evaluation or an earlier data collection period. See Additional File 2 for a codebook for analysis of three health equity domains. Consider innovative analytic techniques to best assess multilevel contributors to implementation success and equity.[65] Step 8. Specify implementation outcomes and evaluate implementation. How to Use the Three Health Equity Domains as Determinants of Equitable Implementation Through formative evaluation to assess barriers and facilitators in each domain,[63, 64] align qualitative interview guides, quantitative measures, and other assessment methods (e.g., participant observation, policy review) to the framework’s determinants—see Page 22/36 Table 1. One might also include patient recipient perspectives in formative evaluation, specifically sampling for those who are experiencing the health or healthcare inequity. We provided an example of an interview guide aligned to domains of the Health Equity Implementation Framework (see additional file 1). Table 1. One might also include patient recipient perspectives in formative evaluation, specifically sampling for those who are experiencing the health or healthcare inequity. We provided an example of an interview guide aligned to domains of the Health Equity Implementation Framework (see additional file 1). Box 1. Application of the Health Equity Implementation Framework to Guide an Equity-Focused Implementation Project: A Case Study Discussion The three health equity domains are key adaptations for implementation scientists and suggested to adapt one’s preferred implementation framework (e.g., CFIR) to incorporate an equity lens. This manuscript, tables, and additional files provide some practical guidance and tools for incorporating three health equity domains into an implementation determinants framework. To focus on health equity explicitly at multiple ecological levels in implementation science and practice will elucidate drivers of health inequities such as structural racism, heterosexism, and patriarchy. Thus, discovery of causes of health inequities should necessitate implementation strategies to overcome or resolve such complex and oppressive structures. Future research should focus on implementation strategies (or other processes) used to address the three health equity determinants of unjust health inequities in our healthcare systems and societies. Implementation science frameworks have been categorized into three types: determinants (establishing what factors determine or predict implementation success), process (clarifying how to address determinants to achieve implementation success), and evaluation (determining metrics and assessment to know when implementation success is achieved).[57] One may also use the Health Equity Implementation Framework in its entirety as an implementation determinants framework, or use the three health equity domains as additions to another implementation determinants framework. For an implementation process framework that incorporates an equity lens, see frameworks proposed by Nápoles and Stewart[12] and Eslava-Schmalbach and colleagues.[60] For an implementation evaluation framework that incorporates an equity lens, see preliminary equity-focused implementation outcomes[60] and the proposed extension of RE-AIM framework.[66] Implementation science frameworks have been categorized into three types: determinants (establishing what factors determine or predict implementation success), process (clarifying how to address determinants to achieve implementation success), and evaluation (determining metrics and assessment to know when implementation success is achieved).[57] One may also use the Health Equity Implementation Framework in its entirety as an implementation determinants framework, or use the three health equity domains as additions to another implementation determinants framework. For an implementation process framework that incorporates an equity lens, see frameworks proposed by Nápoles and Stewart[12] and Eslava-Schmalbach and colleagues.[60] For an implementation evaluation framework that incorporates an equity lens, see preliminary equity-focused implementation outcomes[60] and the proposed extension of RE-AIM framework.[66] There are limitations to our practical guidance presented here. We have piloted test many, but not all, the feasibility and acceptability of steps using three health equity domains described in the results section. Box 1. Application of the Health Equity Implementation Framework to Guide an Equity-Focused Implementation Project: A Case Study Evaluation frameworks can be helpful in selecting implementation outcomes,[26] and it is essential to also select outcomes to evaluate equity in access to, quality of, or outcomes of implementation.[38, 59] The three health equity domains (in addition to typical implementation domains) can be helpful in designing qualitative codebooks or templates for analysis or re-evaluating quantitative measures first collected during formative evaluation or an earlier data collection period. See Additional File 2 for a codebook for analysis of three health equity domains. Consider innovative analytic techniques to best assess multilevel contributors to implementation success and equity.[65] Step 9. Use a framework(s) at micro level to conduct and tailor implementation. As implementation progresses, an implementation plan will need to be tailored as determinants change. The Health Equity Implementation Framework can be useful for determining areas to assess repeatedly and thus, intervene on, throughout implementation. Doing so ensures an equity lens is applied throughout implementation and that implementation processes, such as planning, strategy use, and goal setting, are thoughtfully executed according to dynamic needs. Repeated assessments can be done through multimodal methods, including ones used previously in formative evaluation.[64] Step 10. Write the report. For documenting results of an implementation effort, clarify how the Health Equity Implementation Framework or its three health equity domains were incorporated. For example, barriers and facilitators from formative evaluation may be presented within framework domains. The multimethod assessment and evaluation approaches used in prior steps will provide key information to Page 23/36 Page 23/36 Page 23/36 be reported, making clear why implementation was successful or not, and how certain strategies affected equitable implementation.[13] be reported, making clear why implementation was successful or not, and how certain strategies affected equitable implementation.[13] Discussion We have not piloted feasibility and acceptability of all measures suggested in Table 1. However, we suggest these as starting places, and with confidence, as they all have entire bodies of science showcasing their relevance to health equity. Implementation scientists and practitioners must adopt a health equity lens as foundational to any research-practice gap where inequity exists and collect data on the feasibility, acceptability, and predictive utility of three health equity domains. Abbreviations i-PARIHS: Integrated Promoting Action on Research in Implementation in Health Services U.S.: United States U.S.: United States Conclusion The Health Equity Implementation Framework provided groundwork for the use of implementation science frameworks to address health inequities by incorporating factors that drive health inequities.[11] This manuscript providers practical tools and explanations for systematic integration of health equity and implementation science in frameworks. These tools can assist implementation scientists and Page 24/36 Page 24/36 practitioners to concretely address inequity in implementation across populations to capture and analyze information used to assess health outcomes.[23] practitioners to concretely address inequity in implementation across populations to capture and analyze information used to assess health outcomes.[23] Availability of data and materials Availability of data and materials Data sharing is not applicable to this article as no datasets were generated or analysed during the current study Data sharing is not applicable to this article as no datasets were generated or analysed during the current study Competing interests Competing interests The authors declare that they have no competing interests Funding Funding This work was in part supported by the National Institute of Mental Health (K23MH115100; PI: Dickson). Dr. Singh was also supported by the Department of Veterans Affairs Office of Academic Affiliations Advanced Fellowship Program in Mental Illness Research and Treatment; the Medical Research Service of the Central Arkansas Veterans Healthcare System; and the Department of Veterans Affairs South Central Mental Illness Research, Education, and Clinical Center Declarations Ethics approval and consent to participate Not applicable Consent for publication Not applicable Consent for publication Not applicable Acknowledgements Acknowledgements Drs. Dickson and Woodward are fellows with the Implementation Research Institute (IRI), at the George Warren Brown School of Social Work, Washington University in St. Louis; through an award from the National Institute of Mental Health (5R25MH08091607). We thank Ross Brownson and Sarabeth Broder-Fingert for their continued encouragement and consultation. Thank you to Amber D. Haley for astute and generous consultation on the codebook template. Authors' contributions Authors' contributions ENW conceptualized the manuscript, provided guidance on literature for implementation and health equity domains, prepared and refined all usable tools (additional files), and was a major contributor in writing the manuscript. RSS was a major contributor in writing the manuscript and preparing Tables 1 and 2. PN contributed writing to some implementation and health equity domains, to literature searching and writing in Table 1, and to writing the conclusion. AMC contributed writing to some implementation and health equity domains and to literature searching and writing in Table 1. Page 25/36 KSD helped conceptualize the purpose of the manuscript, prepared and wrote the case study example, and edited writing of the manuscript. JEK refined conceptualizations of the manuscript and edited writing of the manuscript. Acknowledgements References 1. National Partnership for Action. National Stakeholder Strategy for Achieving Health Equity. 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Figure 1 Health Equity Implementation Framework Figures Page 35/36 Figure 1 Health Equity Implementation Framework Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. supplement4.xlsx l 5 d Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. supplement4.xlsx supplement5.docx Page 36/36 Page 36/36	33,057
06/dumas.ccsd.cnrs.fr-dumas-01393296-document.txt_10	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	9,975	16,520	Guillaume Talhouët Capitaine de 50 hommes d'armes Participe au siège de Vitré en 1589 Nommé gouverneur de Redon en 1590 Maître de camp de Mercoeur Participe à la bataille de Craon Dès mai 1594, se détache de Mercoeur en raison de la conversion d'Henri IV Présent aux États ligueurs à Vannes en 1593 Talhouët Guillaume Écuyer, sieur de Keredren Député aux États de Blois en 1588 Capitaine pour la Ligue Présents aux États de Bretagne en 1581 et 1588 Présent aux États en 1591, 1593 et 1594 Talhouët René (de) Écuyer, seigneur dudit lieu Fils de François de Talhouët Présent aux États de Bretagne en 1581 Présent aux États ligueurs en 1592 Fait partie de la délégation envoyée par les États vers don Juan en 1592 (Le) Texier Jean Sieur de la Salle Sénéchal des régaires de Guérande et de la châtellenie d'Escoublac Capitaine de Guérande Procureur-syn dic de Guérande (1591) Député de Guérande en 1591 Tillon Yves Avocat au siège de Vannes États de 1582 Procureur-syndic en 1592 Député de Vannes aux États en 1591-1594 Toulalan Yves Chanoine de Cornouaille, grand chantre et syndic du diocèse de Quimper, recteur de Châteauneuf-du-Faou Député aux États de Blois en 1588 États de 1581, 1582, 1583, 1585, 1586, 1588 Nommé au Conseil d'État et des finances de Mercoeur Député du chapitre de Cornouaille en 1591, 1593 et 1594 Tousselin Jean Chanoine, chantre, scolastique et official de l'église de Nantes Influent soutien de Mercoeur États de 1584, 1585, 1586 Député du chapitre de Saint-Malo en 1591 Mort le 6 octobre 1592 Trousin Guillaume (de) Sieur de Tremeur Noble présent aux tats de Vannes en 1593 Trovausin Guillaume Chanoine de Saint-Pol-de-Léon et recteur de Tremeuchaffin? Député du chapitre de Léon en 1593 Vabarti Pierre Sieur de Guernandrye Procureur-syndic de Quintin Député de la ville en 1594 Vallée Jan (de la) Écuyer, sieur de la Vallée Plumaudan Chevalier de l'ordre du roi Présent aux États de Vannes en 1593 Vauferrier Jean (du) Sénéchal de Dol Député de la ville en 1594 Vieuxchastel Arthur (du) Sieur de Kersal Conseiller au siège présidial de Quimper Sénéchal de Carhaix (1592) Député de Carhaix aux États de Vannes de 1594 Les estatz de ce duche de Bretaigne ayans par serment solennel jure l'ecdit d'unyon suivant les articles redigez en leur assemblee et en la grace de dieu recherche ayde et secours contre les maulx et calamitez qui affligent ce pays ont avecques monseigneur le duc de Mercoeur gouverneur audit duche et soubz l'auctorite duquel ilz sont convoquez en la ville de Nantes sur les cahiers presantez par les trois ordres pour la conservation de la religion, manutention de l'estat du pays et soulaigement du peuple faict les ordonnances cy apres pour estre gardees et observees inviollablement attendant qu'il y est un roy tres crestien et catholique. I. D'autant que la purete en la religion est le fondement de tous estatz et quelle se maintient en l'unite de l'eglise et unyon des catholiques l'ecdit d'unyon du mois de juillet jure aux estatz de bloys derniers sera entretenu et observe comme loy fondamentalle du royaume et les articles sur ce faictes en l'assemblee desdictz estatz de ce pais de Bretaigne jurez par monseigneur de Mercoeur et les deputtez des estatz entre les mains de monsieur l'evesque de Cornouaille y president, seront jurez par tous les catholiques de ce duché par devant les juges ordinaires lesquelz ny les procureurs et greffiers ne prendront aucune chose pour la prestation dudit serment fera le greffier pour l'escripture et pour le regard des ecclesiastiques par devant leur evesque et superieurs de l'eglise. II. Et ceulx de ceste province qui seroient reffuzans de prester ledit serment d'unyon deux mois apres la publication desditz articles sont reputez ennemys du pays et sera procede contre eulx extraodinnairement comme contre perturbateur du repos public et criminelz de leze majeste divine et humaine. III. Sera supplye la sainctete au nom desditz estatz de commettre et deleguer juges qui soient reseans en la ville de Vannes pour juger et terminer les appelations du officiaulx et juges ecclesiastiques et pourvoir aux benefices desquelz sur les reffus des ordinnaires et par droit devollu les provisions apartenant a l'archevesque de tours. IIII. Aussi sera supplie au nom desdictz estatz sa sainctete de pourvoir a la nomination de monseigneur le duc de mercoeur gouverneur et lieutenant general en ce pays et jusques a ce qu'il y ayt ung roy catholique en ce royaume, de personnes de merite et des quallitez requises par les saincts conciles et decretz eveschez, abbayes et benefices destituez de leur evesques abbez et pasteurs lesquelz sont heretiques ou fauteurs d'heretiques. 263 V. Les ecclesiastiques seront maintenu en immunitez, libertez et exemptions qui leur sont concedees tant de droit commun que ordonnances du roy. VI. Et par ce que plusieurs se sont retirez l'eglise contre ladite foy et profession catholique notoirement embrasse le party des heretiques auquel ilz seroient demeurez obstinez juques a presant aucun d'eulx ne sera receu a jurer l'unyon qu'ilz n'aparoisse a monseigneur le gouverneur de sa conversion et profession de foy devant son evesque grand vicaire ou deleguez et ne leur sera faict main levee de leurs biens que par bons effectz ilz n'apparoissent estre catholiques et vrayment de l'unyon et sans que pour ceulx qui ont este mis hors des villes y puissent estre receu. VII. Les blasphemateurs du nom de dieu seront puniz par la rigueur des ordonnaces tant du roy sainct loys qu'autres roys et le duell commun entre les gens de guerre est prohibe et deffendu et en cas de contravention seront les coupables puniz de peines indictes par les ordonnances et ou de eux les appellans et parain de chacun notez infamye et enjoinct aux chefs et capitaines mesmes aux juges ordinaires d'y avoir l'oeil. VIII. Et par ce que de l'administration de la justice deppent le soulagment de la bretz et conservation de l'estat, advenant vaccation d'estatz et offices de judication il y sera pourveu par mondit seigneur de personnes capables et seront lesdittes personnes faictes sans que pour icelles directement ou indirectement se puissent prendre denier et ceulx contre lesquelz sera veriffie audit donner deniers pour lesdictz estatz en seront destituez et privez. IX. Aux estatz et offices tant des courtz souveraines juridictions royalles des finances que tous autres sera pourveu par mondit seigneur lors quelz vacqueront par deces ou autrement. X. Tous magistratz et officiers heretiques ou fauteurs d'heretiques l'ecdit d'unyon sont destituez de leurs estatz offices et dignitez et au lieu d'iceulx sera a l'advenir pourveu par mondit seigneur de personnes catholiques de l'unyon et capables suivant ce qu'il est ordonne cy dessus. XI. Par ce qu'aucuns sieges royauls sont occuppez par les heretiques et leurs fauteurs et a ce que la justice puisse estre administree aux catholiques en lieu certain et en toutte liberte au 264 soulagement desdits catholiques sera pourveu d'un lieu plus commode et de sur acces auquel seront confferez lesdictz sieges royaulx et pour l'execution se pourvoieront par-devers mondit seigneur. XII. L'establissement de translation tant de la court de parlement de sieges presidiaulx monnayes que autres jurisdiction royalles et subalternes unions d'icelles faictes par mondit seigneur des villes tenant le party contraire au villes de l'unyon sont approuvees suivant les lettres du roy et de mondit seigneur. XIII. Les provisions faictes par mondit seigneur des estatz et offices vacants tant par deces que par incapacite des heretiques et leurs fauteurs sont agreables aux estatz. XIV. Est supplye monseigneur le gouverneur de tenir la main a ce que l'auctorite tant des cours souveraines que inferieures soit maintenue, soient les delictz puniz et les jugemens donnez contre les delinquans executez. XV. Les estatz approuvent l'establissement faict par monseigneur d'un conseil d'estat attendant l'assemblee des estatz et trouvant bon le service faict par lesdit conseil et par ce qu'il est necessaire restablir un conseil par l'auctorite de monseigneur et advis des estatz qu'il soit compose des trois ordres ledit seigneur a faict nomination de six et lesdictz estatz de douze qui sont quatre de chaque ordre. XVI. Par ce que le prevost des mareschaulx estably pour purger la province des meschans ne peult a raison des troubles vacquer sy soigneussement a l'exercice de son estat que plusieurs meurtres volleries et autres delictz ne demeurent impuniz pourront les juges royaulx ordinaires des villes esquelles il n'y a siege presidial proche par prevention et proceder a l'instruction et jugement des proces et execution de leurs sentences en dernier ressort appellant tel nombre a juger avecq eulx qu'il est porte par le reglement de la jurisdiction des prevoste des mareschaulx sant que neantmoingn l'attribution jurisdiction puisse estre tirer a consequance et avoir lieu sinon pendant la guerre. XVII. Tous les biens tant immeubles que autres d'heretiques et fauteurs d'iceulx de quelque quallite et condition qu'ilz soient seront saisiz et regiz par main de commissaires lesquelz feront proceder au bail a ferme d'iceuls pour les deniers en provenans estre mis entre les mains du receveur ordinaire et emploiez aux fraiz de la guerre et adjoint aux juges ordinaires 265 commissaires qui seont envoiez par monseigneur a cette fin de l'advis desdictz estatz de proceder a l'execution desdites saisies sans dissiumulation ou connivance dont leurs conscience seront chargees. XVIII. Dons cy devant faictz par mondit seigneur tant du revenu des terres saisies que de deniers a quelque somme a quelques personnes et de quelque nature de deniers que ce soit sont approuvez par les estatz sans que les donnations en puissent estre reserchez. XIX. Et sur les deniers provenant desdictes saisies seront paiees touttes les charges reelles foncieres et hipothecqueres et du benefice saisiz le service faict et les decymes acquitees. XX. Deffances sont faictes a tous capitaines soldatz et autres de ny troubler et empeschez lesdictz commissaires reveveurs et fermiers en la jouissance et perception des fruictz ou revenu desdictes choses saisies sur peine de punition exemplaire, ains au contraire leur prester main forte. XXI. Et pour la diminution et rabais que pouroient pretendre les fermiers pour la non jouissance se pourvoieront par devant les juges ordinaires qui auront faict les saisies pourveu que la ferme n'excede la somme de cens livres et au cas qu'elle excede ladite somme se pourvoieront vers mondit seigneur ou vers le conseil d'estat. XXII. Ne se feront a l'advenir aucuns dons des fruictz et revenu desdictes choses saisies soit eveschez abbayes prieurez et benefices maisons de gentilzhommes et autres ny des greffes de jurisdictions a quelque personnes ou pour quelque cause que ce soyt et sont revocquez tous les dons qui en auroient este faictz cy devant par mondit seigneur a ce que les deniers provenant desdictz fruictz et revenu soient convertiz aux fray de la guerre sauf que mondit seigneur pourra subvenir et pourvoir aux catholiques desquelz seroit le bien occuppe par les heretiques de telles sommes de deniers qu'il jugera necessaire ayant esgard au merite des personnes qui se eront par les mains dudit receveur. XXIII. Et affin de pourveoirs aux plaintes des pauvres laboureurs et villageois qui ont souffert jusques icy tant de pilleries exactions et cruaultez sont faictes deffances a tous gens de guerre et autres sur peine de la vye de prendre lesdictz villageois et laboureurs gehenner tourmenter et en prendre rançon ravir ou enlever leurs bestes de charue et autres bestail instrumens de labeur et tous autres meubles et en cas de contravention sera par le prevost des mareschaulx et 266 concurement par les juges ordinaires informe d'estre delinquans et leur proces faict et parfaict extraordinairement. XXIIII. Et pour plus grand seurete desdictz villageois et laboureurs ceulx qui se trouveront avoir achapte des biens meubles d'iceulx pris et raviz par voye de faict seront condemnez a les rendre et restituez ou la juste valleur d'iceulx et outre punyz d'amande arbitraire. XXV. Deffances sont faictes a tous capitaines soldatz et autres ne prendre prisonniers les femmes que quelque quallite quelles soient ny les enfants soubz l'age de quinze ans et sont des a presant les prises declares nulles. XXVI. Les chasteaux et places fortes qui seront prisent sur l'ennemy seront desmantellees selon qu'il sera necessaire pour l'importance d'icelles. XXVII. Les seigneurs gentilzhommes et autres particulliers de l'unyon qui auront chasteaulx et maisons fortes en feront desmolir les forteresses si mieulx ils n'ayment les conserver et garder a leurs despans sans qu'ilz puissent faire levez ny demander contribution sur les parroisses pour la garde et conservation desdictes places et chasteaulx. XXVIII. Et pour ce que les privilleges de la noblesse oblige les nobles aux armes et que neantmoings plusieurs gentilzhommes prefferant leur commodite a la necessite publicque et manutention de la religion et que plusieurs s'excusent sur l'age et sur le sexe encores qu'ilz possedent terres et fiefz subictz aux armes, seront indiferement tous les nobles capables de porter les armes contrainctz se tenir prestz avecq leurs armes et equipages et les autres fournir hommes suffizant avecq armes et chevaulx chacun selon ses facultez qu'ilz nommeront pour et en fera qu'il sera ordonne par monseigneur le gouverneur marcher soubz la cornette blanche ou autre ine qu'ilz auront chouaisy soubz son bon plaisir. XXIX. Et au cas qu'ilz ne fournoiroient ausdictes armes ilz seront par les juges ordinaires taxez et cottizez de la dixiesme partye du bien dont ils jouiront sans prejuduer neantmoings aux privilleges des nobles du pays ny que cela se puisse tirer a consequance en aucun effect que pour le service de la religion et de l'eglise. XXX. 267 Des nobles et autre faisans profession et faction de gens de guerre seront pour telz compris en touttes les capitulations des villes et chateaux. XXXI. Advenant le deces du noble faisans faction des armes leurs chevaulx et equipaige seront delivrez a l'heritier principal sans que les chefz et capitaines y puissent pretendre aucun droit et pour les non nobles faisant profession des armes les heritiers succederont sinon que les armes et chevaulx eussent este fourniz par les capitaines. XXXII. Les gentilzhommes retirez aux villes de l'unyon seront exemptz des gardes subventions et empruntz qui seront faictz endite villes et tous autres du tiers estat contribueront ausdictes charges. XXXIII. Deffances faictes a tous capitaines des villes et chasteaulx de prendre sur peine de la vye titre et quallite de gouverneur ny de s'attribuer juridiction et ne faire ordonnances lever deniers munitions extraordinaires permettre ou deffandre le trafficq et a ceste fin baillez passeportz et pour iceulx ou autre occasions exiger dans ou dehors les villes sur le peuple aucune chose sans commissions des estatz ou de monseigneur selon les occurances et necessitez des affaires. XXXIIII. Aucun capitaines gentilzhommes ou autre ne pourra tenir deux charges et offices incompatibles et y sera contrainct d'obter dans ung mois apres la publication des presantes. XXXV. Les sauvegardes et passeportz baillez par monseigneur le gouverneur seront inviollablement gardees et contre ceulx qui y attempteront sera procede extraodinairement. XXXVI. Sera a la requeste et dilligence des gens du informe par les juges ordinaires contre ceulx qui auroient de leur auctorite prive faict lever des deniers sur le peuple sans commissions de moinseigneur ou consantements des villes et communaultez pour passe de ce estre procede contreulx ainsi qu'il apatiendra. XXXVII. Et d'aultant que le public a notable interest a la conservation des forestz et boys de haute fustay lesquelz ont este ruinez et depopullez pour la licence que quelques capitaines et particulliers se sont donnez deffances sont faictes a tous capitaines et autres de n'en desmolir 268 coupper et abattre desdites forestz et boys de haute fustaye sur peine de punition corporelle et de tous despans dommaiges et interestz il sera informe contre les contrevenans et les informations envoyees en la court de parlement pour y estre pourveu et ordonne ce que de raison et au cas qu'il seroit necessaire d'abattre desdits boys ne se pourra faire sans commission de monseigneur ou ordonnance des juges royaulx sy la necessite y est lesquelz en feront proces verbales sur l'absence des juges des eaux et forestz et en cas de malversations lesdits officiers en respondront en leur prive nom. XXXVIII. Tous capitaines de marine prenner que prendre commission de monseigneur le gouverneur de faire la guerre sur la mer bailler cauption de quatre mil escuz pour les prises qu'ilz feront. XXXIX. Le nombre des garnisons sera reduict et monseigneur est supplye y donner tel reglement que le pauvre peuple soyt soullage de l'oppression qu'il a cy devant souffert. XL. Et aussy mondict seigneur supplie de remettre la discipline millitaire et affin que les capitaines et soldatz scaichent la justice de ceste guerre en laquelle ilz sont emploiez presteront le serment de garder et observer les articles de l'unyon et pour se faire recoignoistre d'un se sainct party porteront tant les gendarmes que soldtaz croix telle que mondict seigneur ordonnera. XLI. Les maitres de camp et capitaines auront l'oeil que leurs soldatz ne facent aucuns ravaiges et viollences aux personnes et maisons ecclesiasticques gentilzhommes et autres de l'unyon et ou il s'en commettroit aucuns representeront lesdits soldatz au mesme instant que plainte en sera faicte au prevost de camp ou a la justice prochaine des lieulx du delict et tiendront la main a ce que punition s'en soit faicte a faute de quoy en respondront en leurs prives noms. XLII. Et a ce que les ecclesiasticques, gentilzhommes, bourgeois marchans et autre puissent exercer chacun sa vacation en seurete sont faictes deffances aux gens de guerre de prendre aucuns prisonniers du party de l'unyon sur peine de la vye soyt en villes soyt aux champs brusler piller et ravaiger leurs maisons et meubles ny d'user a l'endroit des femmes et filles de force et viollance. XLIII. Aux garnisons des chasteaux ne seront mis gens de cheval sinon qu'il y ayt auculne et particulliere consideration. 269 XLIIII. Les compagnies de chevaulx legers seront composees au plus de cent et au moings de cinquante hommes et aux compaignies de cent hommes n'y aura que quatre chefz et a celles qui ne sont que de cinquante n'y aura que trois et pour ce qu'est des gendarmes observeront les ordonnances et la forme accoustumes. XLV. Les crimes et delictz que commettent les sorldatz seront severement punyz pour reprimer la license et maintenir la discipline millitaire. XLVI. Les compaignies marchans par pays se tiendront serrees pres de leur drapeau et se logeront serrees suivant les ancyenes ordonnances. XLVII. Lors que les monstres se feront, les monstres faictes et auparavant le payement, les ordonnances seront leues aux gens de guerre qui jureront les garder et observer. XLVIII. Les gens de guerre ne pourront contraindre leurs hostes a leur fournir vivres autres que ceulx qui sont en leurs maisons qu'ilz payeront raisonnablement. XLIX. Tous prisonniers de guerre soient gentilzhommes ou capitaines seront declarez de bonne ou mauvaise prinse par mondit seigneur et seront en attendant menez et renduz a la prochaine ville ou garnison de l'unyon sans estre gehen ny tourmentez. L. Pour ce qui est des prinses faictes cy devant sur autres que sur les heretiques ou leurs fauteurs et que n'auroient este jugees bonnes pourront ceulx qui les auront souffertes en leurs personnes et biens en faire informer et pousuivre la reparation en justice. LI. De touttes les rançons declarees bonnes le dixieme sera reserve et employe pour ayder aux fraiz de guerre. LII. Les soldtaz estrangers demeureront soubz leur capitaine et enseigne et deffances a tous capitaines françoys de n'en les retirer ny suborner dautan que soubz ombre de ce se commettent plusieurs insolences et volleryes. 270 LIII. Les soldatz qui auront preste le serment ne se retireront de la charge de leurs capitaines jusques a la monstre prochaine qu'ilz pourront demander leur conge devant le commissaire et controlleur qui ne les pourra estre desnye et s'ilz se retiront autrement et sont reprins seront puniz de mort sans pouvoir estre tenu pour prisonniers de guerre. LIIII. Les monstres des gens de guerre se feront sur les reelles des capitaines qu'ilz presenteront aux commissaires et controlleurs lesquelz seront choisiz et esleu des habitant des villes et communaultez ou seront les garnisons, sans aucun sallaire et par le mesme seront lesdits capitaines ou soldatz paiez a la banque et manuellement par le paieur, presant lesdictz controlleurs et commissaires au desir de l'estat et ordonnance qui en aura este faicte sans que lesdictz deniers passent a l'advenir par les mains des capitaines. LV. Le controlleur a chaque monstre se fera representer le reelle de la derniere et precedante monstre et la cause des absent desnominez en icelle. LVI. Les habitans en chaque ville pourront faire election de luy qui leur sera agreable pour estre sergent maior lequel prestera le serment a monseigneur ou au capitaine de la ville de se porter bien et fidellement en la charge et sans aucuns gaiges. LVII. Et dautant que en affaires des villes et communaultez les habitant d'icelles y ont le principal interest pour leur confirmation des capitaines desdites villes en touttes les affaires qui se presenteront et de leur ressort soient les exploictz de guerre appelleront nombre d'habitant tant de l'eglise de la noblesse que du tiers estat pour les assister et donner advis sur lesdites affaires. LVIII. Ne pourront lesdits capitaines des villes de leur auctorite et sans l'advis et deliberation des magistratz et habitant des villes et communaultez autre que ulx qui auront interest ordonner des fortiffications et demolitions requisent esdites villes et maisons du ressort. LIX. Ne se pourra faire aucune nouvelle imposition sans l'auctorite de monseigneur le gouverneur lequel sera supplier de n'en permettre aucune sans le consentement des estatz de ce pays et ne souffrir que les villes particullieres de ce pays facent imposition sur elles sans le 271 consantement desdits estatz suivant les privilleges et libertez dudit pays sinon qu'en faisant imposition et lever sur eulx capitallement. LX. Les fermes departement et impositions accordez aux estatz de ce pays ne se pourront bailler departer et taxer par les juges de chacune jurisdiction sans y appeler ung ou deux de chaque ordre. LXI. Et a ce que les parroisses ne soyt taxer en divers lieulx et ne confondre l'ordre et forme ancyenne le departement des munitions, charroiz, corvees et nouvelles impositions se feront par les barres et jurisdictions. LXII. Les taxes et impositions qui se feront par cy apres au cas que les estatz advissasent et fust necessaire de faire se feront sur touttes les parroisses sans qu'aucune s'en puisse exempter en vertu d'exemption particulliere, et s'en fera la lever par ung mesme receveur en chaque jurisdiction royalle par les mains duquel passeront tous les deniers qui seront levez. LXIII. Touttes impositions et levees de deniers imposez et soufferte pour la necessite des presant troubles a quelques occasions de quelque nature et condition et en quel lieu elles ayait este introduictz soyt pour fortiffications de ville entretenement de garnisons ou autre suject sont supprinees et ostees sauf a pourveoirs par les estatz a tout ce que sera represante et juge necessaire. LXIIII. Les deniers qui seront par cy apres levez seront emploiez et distribuez par ordonnance de monseigneur suivant l'advis prins par les estatz en ceste assemblee et destination desdictz deniers et a cette fin lesdictz deniers mis entre les mains du tresorier desdictz estatz pour en tenir compte d'an en an en la chambre des comptes presans les deputtez desdictz estatz. LXV. Touttes commissions pour faire levee de deniers lors que par lesdictz estatz aura este trouve bon d'en faire pour corvees et charroiz pour fortiffications et demolitions et pour contribution et munition aux garnisons seront leves et ees aux jurisdictions ou elles sont adressees l'audience tenant et s'en feront les departements par commissaires qui seront prins des trois ordres. LXVI. 272 Les assignations qui se feront sur les deniers de la recepte ordinaire se payeront a l'advenir en la recepte generalle suivant les ordonnances de finances et non par les receveurs particulliers. LXVII. Les departements des corvees qui seront indictes aux parroisses se fera par les capitaines et communaultez et ce feront actuellement par ceulx auquelz elles seront imposer sans qu'ilz ne puissent estre convertiz en deniers et sera donne a chaque homme par jour pour dix huict deniers de pain. LXXIII. Pour ce que la guerre ne se peult faire sans une grande despance le fons de laquelle ne peult este plus assure que en la liberte du trafficq et commerce sera a l'advenir le commerce libre tant par mer que terre en villes rivieres havres et portz et tout autres lieulx de ce duche avecq touttes personnes tant de ce royaume que estrangers sans qu'il soit besoing d'aucuns passeportz sauf des choses deffendues par les ordonnances et pour les bledz, les juges des lieulx prendront garde que les villes en soient fournyes suffizamment et qu'il n'en soyt tire hors le duche par la riviere de loire au pais hault. LXXIIII. Et neantmoings ou se trouveroit quelques ungs faisant trafficq en ceste province saisi de lettres et pacquetz ou qui de pareille intelligence et actions auroyt prejudicie porte scandal ou entreprins contre la saincte unyon et seurete des catholiques d'icelles sera leur proces faict par les juges ordinaires des lieulx et puniz extraodinairement. LXXV. Deffences a tous capitaines ayant charge en terre ou en mer d'attenter sur peine de la vye a la liberte du commerce prendre les personnes arrester les marchandises exiger aucune chose des marchans de quelque estat ou quallite qu'ilz puissent estre et sont touttes privses faictes desdictes personnes et desdictes marchandises declarees nulles faisant lesdits marchans aparoistre par lettres d'advys ou de charte partye sans fraude que lesdites prises estoient menees conduictes aux villes de l'unyon. LXXVI. Les rempartz des costes seront remparez les guetz et fay y entretenu et enjoinct aux capitaines et commissaires des bans et arriere bans ou il n'y auroit capitaines des costes commettre personne en chaque parroisse pour contraindre les habitans suictz auxdictz guetz. LXXVII. Exemples de procuration ADIV, C 3189, Procuration de la ville de Fougères – 1591 Extraict des registres de l' hostel de ville et conseil estably a foulgere s par monseigneur le duc de mercoeur et de penthievre gouverneur general en bretagne. Sur requeste du procureur des bourgeoys et habittans dudit foulgeres a este faict lecture des lettres de mondict seigneur donnees a vannes le dixhuitiesme jour du presant moys de luy soubz signees par lesquelles sa grandeur declare a ceste communaulte la convocation des estatz generaulx de ce royaulme se debvoir faire le vingt cinquiesme du prochain en la ville d'orleans et afin que ceulx qui seront deputtez de ceste province soyent mieux instruictz et informez de ce qu'ilz auront a representer et leurs cayers plus meurement dressez et arrestez mondict seigneur auroit assigné la convocation des estatz de ceste province au douziesme prochain en la ville de nantes avecq commandement expres ausdictz habittans de commettre et deputter aulcuns d'entreulx pour se rendre audit nantes avecq memoires instructions et tout pouvoir a ce requis et ladicte lecture faicte a este par l'advys et commun consentement desditz bourgeoys et habittans commys et deputte Pierre lebigot sieur du breil procureur syndic de ceste communaulte qu'il a este esleu et choisy par lesditz habittans comme celuy d'entreulx qu'ilz cougnoissent aultant bien instruict et cougnoissant des affaires zele et affectionne a la manutention de nostre religion bien et conservation du pays que aulcun des autres habittans auquel ladite communaulte a donne pouvoir de comparoir en ladite assemblee d'estatz et de faire gerer negotier consentir et accorder tout ce que veoyra propre pour le bien du pays et de ceste communaulte en particulier avecq requisitoire a noz seigneurs desdits estatz de vouloir ouir entendre et recepvoir leurdit procureur et deputte promectans et ont jure lesdits habittans avoir aggreable tout ce que sera par luy gere negotye et consenty en leur nom ausdictz estatz tout ainsy que sy ladicte communaulté bourgeoys et habittans dudict foulgeres estoyent presens combien que l'affaire requere mandement plus special ou proscure de personne faict audict conseil le vingt neufviesme jour de janvier l'an mil cinq cens quatre vingtz et unze. Le quinziesme jour de mars mil cincq centz nonante trois en l'assemblee des gentilzhommes de l'evesche de Leon tenue en la ville de Lesneven soubz l'auctoritte de son altesse et en vertu de ses lettres pour la deputation des estatz assignez a tenir a Vanes les vingtiesme dudit mois ladite assemblee d'ung commun advis et consentement oultres les aultres gentilzhommes dudit evesche qui se pourront trouver ausdits estatz a nomme pour s'y trouver avecq tout pouvoir recquis et pertinent en tel cas accoustume nobles gens Hamon de Kergadiou sieur dudit lieu, Bernard Guernisac sieur de Kerchain. L'ung et chascun d'eulx faict gree et consenty en la presence de messieurs le seneschal et procureur du roy de la court et seneschausee dudit Lesneven et du greffier d'ycelle court et ce d'autoricte d'icelle soubz le signe dudit greffier er des soubzsignantz nottaires royaulx a la requeste de ladite assemblee lesdits jour et an. Les gens des troys estatz de ce pays et duché de Bretaigne convoquez et assamblez en la ville de Vennes soubz l'auctorite de monseigneur le duc de Mercoeur et de Penthievre, pair de France, prince du Saint-Empire et de Martigues, gouverneur de Bretaigne pour deliberer et adviser des affaires de la province et des moiens de remedyer aux maux et calamitez ausquellles elle est submergee par la faction des heretique et leur faulteurs, licence et dereglement des gens de guerre, aiantz sur le tout pris avis et resolution a l'honneur de dieu, obéissance du saint siege, service de monseigneur et soullagement du pauvre peuple. Supplient mondit seigneur le gouverneur de les maintenir tousiours par sa force et auctorite contre les heretiques, les conserver en l'obéissance qu'ilz doibvent au saint siege de laquelle ilz ne veullent jamais se departyr non plus que de la couronne de France, ni ne souffrir que ce face autre exercice de la religion en ceste province que de la religion catholique appostolique et romaine, ny que les libertez et franchises du pays ne soient aulcunement dyminuez ny alterez, promettant a cette fin demeurez uni avecq mondit seigneur et employer vye et moiens. Supplient aussi mondit seigneur voulloir embrasser le soullagement du pauvre peuple afflige et ruyne d'une longue guerre, lequel ne respire appres dieu qu'en sa vertu et bonte et pour cest effect embrasser les occurances et occasions que se pourront presenter d'asseurer le repos et la tranquillite de cette province a ce que la religion conserver elle puisse ce remettre des ruynes souffertent et la guerre. Supplient mondit seigneur avoir agreable les depputez qu'ilz ont choisy et nommez pour 'assister et luy donner advis lors que sa prudence jugera estre necessaire et le prandre en la direction des affaires. Et d'aultant que la seurte du pays et des villes ne peult estre sans entretenir grand nombre des gens de guerre tant aulx garnisons qu'a la campaigne subvenir aulx magasins et munitions, payer les gages des officiers, fournir aux voaiges inopinez, mesmes au payement du plat de mondit seigneur, soldes de ses gardes et autres infiniz fraictz qui suivent la guerre. 279 Les estatz ont advise d'accorder la levee des fouages et impostz et billotz, de continuer la pancarte et la subvention le tout pour un an seullement ainsi et de la manière quelle fut arrester en l'assemblee des estatz tenuz en cette ville de Vannes en l'annee quatre vingtz treze sauf un cinquieme qu'ilz ont diminue de ladite pancarte pour acommoder [] du commerce, et supplient monseigneur en consideration de la continuation dudit fons ne bailler aucunes coummissions extraordinaires pour lever deniers sur le peuple, ny mesmes pour la lever des franz archers et esleuz de laquelle ilz demeurent pour cette annee deschargez comme aussi d'oster plusieurs petites garnisons qui ne servent qu'a piller et ravaiger le pauvre paisan et luy oster le moien de fournir aulx subventions qui luy sont imposez et ordonnez comme aussi ilz le supplient de revoquer touttes commisions qu'il auroit cy devant baillees pour lever deniers et commander justice exemplaire estre faicte de plusieurs volleryes et pilleries que font plusieurs gens de guerre et autres a ce que par l'exemple de la punition un chacun soit retenu de mal faire. Supplient aussi londit seigneur de decerner commission et faire commendement a tous juges et officiers de l'union d'informer sans aucunes connivances ou dissimulation des levees de deniers et autres contributions exiger du peuple sans commissions de mondit seigneur, mesmes des sacrilleges brullement et viollement qui ont este commises ny aiant meilleur moien d'assister et secourir le publicq que par la recherche desdites exactions et faisant faire la justice. Supplient monseigneur d'avoir agreable l'offre qu'ilz lui font d'emploiez leur vye et moiens pour la deffance de la religion catholique et romaine pour le bien du pays et de service. Fait en l'assemblee desdits estatz tenuz a Vennes au pallaye royal dudit lieu le douziesme jour de ce may MVC IIIIXX XIIII. Charles de Bourgneuff de Chronologie 12 juillet 1575 : Mariage Philippe Emmanuel de Lorraine et Marie de Luxembourg 5 septembre 1582 : Mercoeur nommé gouverneur de Bretagne 10 juin 1584 : mort de François d'Alençon duc d'Anjou 31 décembre 1584 : traité de Joinville entre les Guises et Philippe II d'Espagne . Mercoeur n'a pas en voy é de mandataire. 1585 30 mars : Manifeste de Péronne : justification prise d'armes ligueuses 7 juillet : traité de Nemours entre Henri III et les princes catholiques. Mercoeur reçoit deux places de sûreté en Bretagne : Dinan et Concarneau. Été : début de la huitième guerre de Religion, opérations dans le Poitou. 9 septembre : excommunication par le pape d'Henri de Navarre 1587 20 octobre: bataille de Coutras, victoire d'Henri de Navarre. 24 novembre : bataille d'Auneau, duc de Guise défait les reîtres allemands appelés en renfort par les protestants. 1588 12 mai: journée des Barricades à Paris. Henri III quitte la capitale pour Tours. 15 juillet : signature de l'édit d'Union 18 octobre : ouverture des États généraux à Blois 23 et 24 décembre : assassinat du duc de Guise et de son frère le cardinal de Guise. 1589 2-5 mars : arrestation de Claude Faucon Ris, premier président du Parlement de Bretagne sur ordre de Mercoeur. 13 mars : journée des Barricades à Rennes. 15 mars-21 mars : Mercoeur à Rennes. 22 mars : Mercoeur prend Fougères en achetant le lieutenant du château. 23 mars-14 août : siège de Vitré par les ligueurs, mais la ville reste aux royaux. 4 avril : Rennes revient au roi. 7 avril : installation municipalité ligueuse à Nantes. 13 avril : arrêt du Parlement de Bretagne met Mercoeur hors-la-loi. 18 avril : Mercoeur destitué de sa charge de gouverneur par Henri III 30 avril : entrevu de Plessis-lès-Tours entre Henri III et Henri de Navarre. Mai : Mercoeur s'empare de Châteaubriant. 1er juin : ligueurs capturent le comte de Soissons désigné pour remplacer Mercoeur 7 juin : prince de Dombes nommé à la place de Mercoeur par le roi. 2 août : mort d'Henri III 281 4 août : ligueurs proclament le cardinal de Bourbon roi de France sous le titre de Charles . 13 août : arrivé de Dombes à Rennes, fait lever le siège de Vitré le 14. 11 septembre : Parlement de Rennes reconnait Henri IV, un des premiers à le faire. 28 septembre : Quimper passe à la Ligue. Septembre : sac de Tréguier par des paysans menés par des gentilshommes ligueurs, mise en place conseil de la Sainte-Union à Morlaix, Brest au roi. 9 décembre : Henri IV à Laval. 13 décembre : Châteaubriant reprise par les royaux. 1590 8 janvier : Mercoeur installe un Parlement ligueur à Nantes. fin février : échec de la tentative de Dombes pour s'emparer d'Ancenis. 7-8 mars : ligueurs reprennent Châteaubriant. 11 mars : Malouins s'emparent du château, mort du gouverneur de la ville. 14 mars : bataille d'Ivry, victoire d'Henri IV contre Mayenne. 2 mai : prise d'Hennebont par les royaux, puis peu après de Quimperlé. 9 mai : mort du Cardinal de Bourbon dans sa prison. 11 juin : Mercoeur prend Blavet. 4 septembre : raid des royaux sur Carhaix. Septembre : attaque du château de Roscanou. 12 octobre : débarquement à Saint-Nazaire des premières troupes espagnoles. Novembre : nouveau sac de Carhaix. 27 décembre : ouverture à Rennes des États royaux de Bretagne. Les premiers depuis le début du conflit. 31 décembre : Mercoeur reprend Hennebont avec l'aide des Espagnols. 1591 21 mars : ouverture des États ligueurs à Nantes. 4 avril : accord Henri IV et États de Bretagne avec Elizabeth d'Angleterre pour aide militaire. 10 avril : clôture des États ligueurs. Début juin : royaux avec l'aide des Anglais s'emparent de Guiguamp. 23 juin : « bataille » du Marc'hallac'h. 18 juillet : mort de François de la Noue, royaux lèvent siège de Lamballe Novembre : Gabriel de Goulaine prend Blain pour la Ligue. 1592 Janvier : Mercoeur s'empare d'Ingrandes et Candé. 21 mars : ouverture des États ligueurs à Vannes. 9 avril : clôture des États ligueurs. 23 mai : bataille de Craon, victoire de Mercoeur et des Espagnols sur Dombes et les Anglais. 20 août : maréchal d'Aumont nommé en Bretagne à la tête de l'armée royale en remplacement du prince de Dombes. 28 décembre : ouvertures des États royaux à Renn 1593 26 janvier-8 août : États généraux de la Ligue à Paris. 13 avril : ouverture des États ligueurs à Vannes. Échec d'une embuscade des royaux contre Mercoeur qui s'y rend. 29 avril-7 mai : conférence de Suresnes entre Henri IV et les députés des États ligueurs. 5 mai : clôture des États ligueurs. 25 juillet : abjuration d'Henri IV à Saint-Denis 31 juillet : trêve générale entre Henri IV et Mayenne. 14 août : trêve publiée à Rennes. Mercoeur rechigne à l'appliquer. 28 août : Mercoeur accepte la trêve. 18 octobre : ouverture des États royaux. 1er novembre : prolongation trêve jusqu'au 1er janvier. Décembre : arrivée de renfort espagnol. 1594 27 février : sacre d'Henri IV à Chartres. Début mars : premiers ralliements de gentilshommes bretons à Henri IV. 22 mars : Henri IV entre dans Paris. Fin mars : Espagnols commencent construction du fort de Crozon à la pointe de Roscanvel achevé en 4 mois. 2 mai : ouverture des États ligueurs à Vannes. 12 mai : clôture des États ligueurs. Derniers réunis pour la Ligue. 8 juillet : Mercoeur à Ancenis, discussions avec sa soeur la reine Louise qui l'encourage à se soumettre. 9 août : traité du Folgoët : soumission du Léon. Juillet-août : soumission de Lezonnet, gouverneur de Concarneau. 25 août : soumission de Morlaix. 4 octobre : édit de réduction de Saint-Malo. 9-21 octobre : maréchal d'Aumont assiège victorieusement Quimper. 17 novembre : royaux s'emparent du fort de Crozon. 20 novembre : premier serment de Mercoeur sur les droits de l'infante en Bretagne. 12-21 décembre : conférences d'Ancenis. 1595 Début de l'année: évêchés de Tréguier, Léon et Cornouaille dans l'obéissance du roi. 17 janvier : Henri IV déclare la guerre à Philippe II. 23 janvier : ouverture des États royaux à Rennes. 20-20 mars : reprise des conférences d'Ancenis. 5 juin : victoire d'Henri IV à Fontaine-Française. Début juin : soumission de François de Talhouët, gouverneur de Redon. 30 juin : second serment de Mercoeur sur les droits de l'infante. 19 août : décès du maréchal d'Aumont, blessé le 3 juillet devant Comper. 283 Août : soumission d'Urbain de Laval Boisdauphin. 17 septembre : absolution d'Henri IV par le pape Clément VIII. 12 octobre : publication trêve de trois mois entre Henri IV et Mayenne. 20 novembre : ouverture des É royaux. Novembre : traités de Folembray, soumission du duc de Mayenne, puis en janvier 1596 de Nemours et Joyeuse. Début 1596 : la Ligue ne tient plus que douze villes situées en Haute-Bretagne : Hennebont, Vannes, Pontivy, Josselin, Saint-Brieuc, Lamballe, Dinan, Dol, Fougères, Châteaubriant, Guérande et Nantes. 22 mars : accord sur les trêves. Avril : reconduction des trêves jusqu'au 1er juillet. 2 juillet : Brissac nommé lieutenant général en Bretagne. 30 septembre : Mercoeur reçoit semonce du pape le poussant à se soumettre à Henri IV. 15 octobre : reprise des pourparlers à Chenonceaux, Tours et Ancenis. 9 décembre : ouverture des États de Bretagne à Rennes. Trêves reconduites jusqu'au 1er avril 1597. 1597 17 mars : rupture des négociations. 15 avril : royaux reprennent Châteaubriant. Début juin : mutinerie à Blavet. Début août : renfort espagnol. 17 novembre : reconduction des trêves jusqu'au 1er janvier 1598. Archives départementales Ille-et-Vilaine - Série B 1 Ba 11 : Registres des enregistrements du Parlement de la Ligue à Nantes. 1 Bb 75 : Registres secrets du Parlement de la Ligue de Nantes, séance novembre 1590. 1 Bb 437-439 : Registres secrets du Parlement de la Ligue à Nantes, 1591-1594. 1 Bf 1621 -1625 : Arrêts sur rapport du Parlement de la Ligue de Nantes. Archives départementales Ille-et-Vilaine - Série C États royaux : C 2643 : Registr e compren ant copie des procès-verbaux des États de Rennes en 1588, 1590, 1592, 1593. C 2644 : Procès-verbaux des États de Rennes de janvier 1595 et novembre 1595. États ligueurs : C 3187 : • Inventaire • PV État 1591 C 3188 : • Serment des ligueurs 1591 • Cahier des ordonnances des États 1591 C 3189 : • Procuration députés des villes • Procuration députés clergé • Requêtes particulières C 3190 : • Parchemin convocation des États provinciaux par Mercoeur • Réponse et conclusion des États • Requête des villes 285 C 3191 : • Gratification, aumônes demandées par les religieux • Députation envoyée à Don Juan d'Aiguila et Don Diego, commandant de l'armée espagnole 1592 C 3192 : • Pancartes et baux des devoirs 1591 • Taxes diverses • Cautionnement du trésorier des États • Lestage, délestage et pilotage des navires entrant en Loire • Dépenses pour la guerre 1591 C 3193 : Procès-verbaux des délibérations des États 1592 C 3194 : • Supplications des États à Mercoeur • Procuration des députés de l'Église 1592 • Procurations des députés du tiers 1592 • Liste des membres des États 1592 C 3195 : • Requête pour secours Quimperlé • Baux des devoirs et impôts et instruction aux officiers des principales villes touchant la levée des devoirs • Documents enquêtes vol du commis à la recette de Penmarch Yvon Longes • Enquête contre La Fontenelle 21 mars 1592 • États des gages des officiers des États • État de la recette et la dépense que fera le sieur Loriot, trésorier des États C 3196 : Procès-verbal des délibérations des États tenu à Vannes du 13 avril au 5 mai 1593 C 3197 : • Procurations des députés du clergé 1593 • Procurations des députés du tiers 1593 • Rançon du duc d'Elbeuf baron d'Ancenis • Remontrances • États des fonds, gages, recette de la pancarte Devoirs, impôts et billots • Instructions données aux députés envoyés aux États généraux convoqués au 17 janvier 1593 à Paris C 3198 : • Procès-verbal des délibérations des États tenus à Vannes du 2 au 12 mai 1594 • Minutes des délibérations C 3199 : • Procurations des députés de l'Église 1594 • Procurations du tiers 1594 • Mémoires, articles et remontrances des villes C 3200 : • Remontrances au duc de Mercoeur et commission générale 1594 286 • Délibérations diverses • Requêtes et ordonnances de paiement de frais de voyage pour députés États généraux de 1593 • Instructions aux députés envoyés vers Don Juan 1594 • Instructions pancarte • Examens des comptes du trésorier • Requêtes aux États par divers particuliers C 3201 : Compte des États 1591-1597 C 3202 : • État recette de Jean Loriot • État des gages des officiers • Ordonnances et quittances C 3203-3205 : Ordonnances et quittances des paiements faits par le trésorier des États C 3744 : Documents relatifs au procès des États contre Jean Loriot, trésorier des États de la Ligue • État de la recette et dépense de Jean Loriot en 1591-1592 • État de la recette et dépense de Jean Loriot en 1593-1594 • État de la recette et dépense de Jean Loriot en 1594-1595 • Bref calcul de la recette et dépense de Jean Loriot en 1597-1598 • Taxations des droits de recettes du comptable pour 1591-1592 • Taxations des droits de recettes du comptable pour 1592-1593 • Taxations des droits de recettes du comptable pour 1593-1595 • Taxations des droits de recettes du comptable pour 1595-1598 • Frais et vacations des commissaires qui participent à l'examen des comptes du trésorier • Copie d'une ordonnance de paiement de Mercoeur au trésorier datant de 1596. 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TABLE DES ANNEXES Annexe 1 Carte du clergé aux États ligueurs 1591-1594 227 Anne xe 2 Graphique de l'origine géographique des députés du clergé 227 Annexe 3 Tableau de la présence du clergé aux États ligueurs de 1591 à 1594 228 Annexe 4 Tableau de la présence des villes aux États ligueurs de 1591 à 1594 229 Annexe 5 Carte des villes présentes aux États de 1591 à 1594 230 Annexe 6 Graphi que du nombre de députés envoyés par chaque ville entre 1591 et 1594 230 Annexe 7 Carte répartition par évêchés des principales possessions des nobles présents aux États ligueurs 231 Annexe 8 Tableau des avants noms des nobles 231 Anne xe 9 Graphique répartition par année des requêtes reçues aux États 232 Annexe 10 Graphique répartition par évêchés d'origine des requêtes reçues par les États 232 Anne xe 11 Graphique c atégories socio-professionnelles des requérants 233 Annexe 12 Graphique nombre de requêtes par villes et paroisses 233 Annexe 13 Carte de l'origine par évêché des requêtes 234 Annexe 14 Carte de l'origine par paroisses et villes des requêtes aux États ligueurs 234 Annexe 15 Tableau des thèmes des requêtes 235 Annexe 16 Cartes des garnisons ligueuses en Bretagne pour 1593 236 Annexe 17 Carte des garnisons ligueuses en Bretagne pour 1591 236 Annexe 18 Tableau estimation de la recette du trésorier des États pour un an 239 Annexe 19 Tableau du revenu théorique des États de 1591 à 1597 237 Annexe 20 Tableau deniers non reçus par les États de 1591 à 1597 237 Annexe 21 Tableau recette du trésorier des États par évêchés de 1591 à 1597 238 Annexe 22 Graphique participation des évêchés à la recette des États de 1591 à 1597 238 Annexe 23 Tableau recettes et dépenses des États 238 Annexe 24 Table répartition des dépenses dans le compte des États de 1591 à 1597 239 Annexe 25 Tableau fermier des devoirs des États ligueurs 240 Annexe 26 Tableau offres pour les fermes des devoirs des États 240 Annexe 27 Tableau recettes des devoirs dans les comptes de Jean Loriot 241 Annexe 28 Tableau de la valeur des fermes par évêchés de 1591 à 1595 242 295 Annexe 29 Graphique origine géographie de la recette des devoirs pour 1592-1593 243 Annexe 30 Graphique origine géographique de la recette des devoirs pour 1593-1594 243 Annexe 31 Tableau de la répartition par évêché de la subvention de 73000 écus en 1592 244 Annexe 32 Graphique de la valeur (en écus) et répartition par évêché de la subvention pour 1593-1594 244 Annexe 33 Tableau des dépenses faites par le trésorier des États en 1591-1592 274 Annexe 34 Tableau des dépenses non financées 1591-1592 245 Annexe 35 Graphique principaux postes de dépenses en 1591-1592 245 Annexe 36 Tableau des dépenses 1593-1594 246 Annexe 37 Graphique répartition des dépenses en 1593-1594 246 Annexe 38 Tableau des dépenses 1594-1594 247 Annexe 39 Graphique répartition des dépenses en 1594 -1595 247 Annexe 40 - Copie d'une ordonnance de paiement du duc de Mercoeur au trésorier des États 249 Annexe 41 - Copie d'une ordonnance de paiement des États 250 Annexe 42 - Notices biographique des députés aux États ligueurs 251 Annexe 43 - Cahier des ordonnances des États – ADIV C 3188 263 Annexe 44 Exempl es de procuration 276 Annexe 45 Réponse des États au duc de Mercoeur - 1594 279 Annexe 46 Chronologie 281 296 TABLE DES MATIÈRES Re merciements 3 Sommaire 5 Abréviations 7 Introduction 9 Partie I – Le fonctionnement des États ligueurs 21 Chapitre 1 – Assembler les États 23 A.	18,183
https://openalex.org/W2473593485	OpenAlex	Open Science	CC-By	2,016	Morphological State of Rivers in the Ilova River Catchment	Mladen Plantak	Croatian	Spoken	11,147	24,052	HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Original scientific paper Izvorni znanstveni članak UDK 911.2:551.4(282.249 Ilova) 551.444(282.249 Ilova) Original scientific paper Izvorni znanstveni članak Original scientific paper Izvorni znanstveni članak Mladen Plantak Ivan Čanjevac Iva Vidaković Primljeno / Received 19-02-2016 / 2016-02-19 Prihvaćeno / Accepted 14-03-2016 / 2016-03-14 Primljeno / Received 19-02-2016 / 2016-02-19 Prihvaćeno / Accepted 14-03-2016 / 2016-03-14 HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Introduction With accession to the European Union (EU) and the already prior acceptance and progressive implemen tation of the Water Framework Directive (WFD), Cro atia undertook the commitment of monitoring and as sessment of the hydromorphological status of its waters. A good hydromorphological status has been recognised at EU level as one of the most important preconditions for achieving a good biological and physicochemical status (WFD, 2000; Feld, 2004). For this reason, hydro morphological quality elements are included in the clas sification of the ecological status of water bodies pur suant to the WFD, together with biological and phys icochemical quality elements. The hydromorphological status fundamentally encompasses the hydrological re gime, river continuity and the morphological conditions (MEANDER, 2013). In recent times, especially after adoption of the WFD, the concept of hydromorphology is considered as an interdisciplinary field incorporating hydrology and (fluvial) geomorphology (Belleti et al., 2015). In hydromorphology, the emphasis is placed on hydrological and morphological features and processes in running waters as a starting point for quality man agement and restoration of rivers (Newson and Large, 2006). The complexity of processes and interrelations in and around rivers creates a necessity for a broader knowledge of ecology, pedology and hydro-engineering in hydromorphological research (Lewandowski, 2012; Bonacci, 2015). Ulaskom u Europsku uniju i već prethodnim prihvaćanjem i postupnom implementacijom Okvirne direktive Europske Unije o vodama (ODV, 2000) Hrvatska je preuzela obvezu pra ćenja (monitoringa) i ocjene hidromorfološkog stanja vodotokâ. Na razini zemalja Europske unije uvidjelo se da je za dobro biološko i fizikal no-kemijsko stanje vodotokâ jedan od najvažnijih preduvjeta njegovo dobro hidromorfološko stanje (ODV, 2000; Feld, 2004). Zbog toga je hidro morfološko stanje uključeno u ukupnu ocjenu ekološkog stanja vodnih tijela prema ODV-u kao element ravnopravan biološkom i fizikalno-ke mijskom stanju. Hidromorfološko stanje u osnovi obuhvaća hidrološki režim, neprekinutost (konti nuitet) toka i morfologiju korita (MEANDER, 2013). U novije se vrijeme, a posebice nakon do nošenja ODV-a, pod pojmom hidromorfologija smatra interdisciplinarno područje koje povezuje hidrologiju i (fluvijalnu) geomorfologiju (Belleti i dr., 2015). U sklopu hidromorfologije naglasak je na hidrološkim i morfološkim obilježjima i pro cesima tekućica kao polazištu kvalitetnog uprav ljanja i revitalizacije vodotokâ (Newson i Large, 2006). Zbog kompleksnosti procesa i odnosa u tekućicama i oko njih često su za hidromorfološ ka istraživanja potrebna šira znanja iz ekologije, pedologije i hidrotehnike (Lewandowski, 2012; Bonacci, 2015). Morfološko stanje tekućica u poriječju Ilove The Ilova River catchment in Central Croatia is un der significant human impact as a result of extensive river training works, construction of canals and fish farms. The consequences of building these hydrotechnical struc tures and river training works on the hydromorphology of streams in the Ilova catchment have not been investi gated to date. Following a historical overview of the sig nificant water works, this paper depicts the results of an assessment of morphological features of streams in the Ilova catchment pursuant to the EU Water Framework Directive (WFD) requirements and Croatian Waters’ Methodology for monitoring and assessment of hydro morphological features. Poriječje rijeke Ilove u Središnjoj Hrvatskoj pod znatnim je antropogenim utjecajem zbog brojnih hidro tehničkih radova, izgradnje kanalâ i ribnjakâ. Do sada u poriječju Ilove nisu istraživane posljedice izgradnje tih hidrotehničkih objekata i uređenja vodotokâ na njihovu hidromorfologiju. Nakon povijesnog pregleda značajnijih hidrotehničkih radova u radu su prikazani rezultati istra živanja morfoloških značajki vodotokâ u poriječju Ilove prema smjernicama Okvirne direktive EU-a o vodama i Metodologije monitoringa i ocjenjivanja hidromorfološ kih pokazatelja Hrvatskih voda. Analizirana su ukupno 534 km vodotokâ i određeno je 48 vodnih tijela za koja je utvrđeno morfološko stanje. Ukupno 38,5 % duljine vodotokâ, odnosno 21 vodno ti jelo u poriječju Ilove, ne zadovoljava ciljeve prema Okvir noj direktivi EU-a o vodama jer je dobilo ocjenu umje reno promijenjeno, loše ili vrlo loše morfološko stanje. Zabrinjavajuće je da su loše ocjene, uz dionice u naselju i oko njega, dobile i mnoge dionice izvan naselja. A total length of 534 km of streams was analysed, which led to the delineation of 48 water bodies for which the morphological condition was evaluated. In the Ilova River catchment, 38.5% of the total watercourse length, respectively twenty-one water bodies, did not meet WFD objectives as they were classified as moderate, poor or bad with respect to morphological condition. The results showing that not only those water bodies within and around settlements but also remote water bodies have a failing morphological status is a matter of concern. Ključne riječi: hidromorfologija, morfologija, Okvirna direktiva EU-a o vodama, antropogeni utjecaj, vodno tijelo, Ilova Key words: hydromorphology, morphology, EU Water Framework Directive, water body, anthropogenic influence, Ilova River 5 HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Introduction In Croatia, hydromorphological research and re search on the effects of water works on the hydrology and morphology of rivers is scarce. Only recently, in January 2015, the document Methodology for monitoring and assessment of hydromorphological features (Methodol ogy, 2015) was published. The morphological assessment presented in this paper is based on that methodology issued by Croatian Waters. Publishing of the Method ology is certainly an incentive for hydromorphological research and a stronger inclusion of hydromorphology into water management and protection of river systems of Croatia. U Hrvatskoj ima nedovoljno hidromorfoloških istraživanja te istraživanja posljedica hidrotehnič kih zahvata na hidrologiju i morfologiju tekućica. Tek je nedavno, u siječnju 2015., objavljen doku ment Metodologija monitoringa i ocjenjivanja hi dromorfoloških pokazatelja (Metodologija, 2015). Metodologiju su objavile Hrvatske vode te se na njoj temelji morfološko ocjenjivanje u ovome radu. Njezino izdavanje svakako je poticaj za hidromor fološka istraživanja i snažnije uključivanje hidro morfologije u sustav upravljanja i zaštite riječnih sustava Hrvatske. The area of research in this paper encompasses the river system of the Ilova River in Central Croatia (Fig. 1). In geographical scientific literature, the Ilova Riv er catchment is rather neglected, especially regarding water resources or river system analyses. To date, there has been no published scientific paper from the fields of hydrogeography, hydrology or hydrogeology system Istraživano područje ovoga rada obuhvaća riječ ni sustav rijeke Ilove u Središnjoj Hrvatskoj (sl. 1). U geografskoj znanstvenoj literaturi poriječje Ilove prilično je zanemareno, posebice sa stajališta ana lize vodnih resursa ili riječnog sustava. Do danas 6 atically dealing with this area, although it is especially interesting and complex from these standpoints. The hydrology of the Ilova River has been studied in pa pers dealing with the wider Sava River catchment or the basin of the Danube River in Croatia (Čanjevac, 2012; Barbalić and Kuspilić, 2014; Čanjevac and Orešić, 2015). Thereby, the objectives of this paper are twofold; to initiate hydromorphological research in general on one hand, and regional hydrogeographical research of less studied regions of Croatia on the other. nije objavljen nijedan znanstveni rad iz područja hidrogeografije, hidrologije ili hidrogeologije koji bi se sustavno bavio tim područjem, premda je ono s tih stajališta izuzetno zanimljivo i složeno. Ilova je s hidrološke strane dijelom analizirana u radovi ma koji su se bavili širim prostorom poriječja Save ili hrvatskog dijela poriječja Dunava (Čanjevac, 2012; Barbalić i Kuspilić, 2014; Čanjevac i Orešić, 2015). Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove Introduction Stoga su ciljevi ovoga rada dvojaki. S jedne strane, želi se potaknuti hidromorfološka istraži vanja općenito, a s druge regionalna hidrogeograf ska istraživanja do sada manje istraženih područja Hrvatske. Morfološko stanje tekućica u poriječju Ilove The Ilova River is the left tributary of the Sava River (Fig. 1). The spring of the Ilova River is located on the northern slopes of Papuk Mountain at Velika Babina Gora (Đulovac municipality) at an altitude of 205 m. 800 m downstream it receives water from two smaller springs (identified on topographic map 1:25,000) and continues through the valley, which is furthermore con ditioned by the fault between Papuk Mountain and the southern slopes of Bilogora and takes on a north-eastern towards south-west flow path. Three locations can be considered as the river mouth of the Ilova River (Plan tak, 2014). The first location is the end of the natural river channel of Ilova River into Stari Trebež River lo cated 5.5 km south of the village of Kutinsko Selo. The second location is the river mouth of the Pakra River into Ilova River and the third location is the present-day Rijeka Ilova lijevi je pritok rijeke Save (sl. 1). Izvire na sjevernim obroncima Papuka u naselju Velika Babina Gora (općina Đulovac) na nadmor skoj visini od 205 metara te 800 metara nizvod no prima vodu iz dva manja izvora (identificirani pomoću TK 25 000), nastavlja se dolinom koja je nadalje uvjetovana rasjedom između Papuka i juž nih obronaka Bilogore te poprima smjer otjecanja od sjeveroistoka prema jugozapadu. Za ušće Ilove mogle bi se uzeti tri lokacije (Plantak, 2014). Prva je lokacija ušće prirodnoga korita Ilove u Stari Trebež, koje se nalazi 5,5 km južno od naselja Ku Sl. 1. Poriječje Ilove Fig 1 The Ilova River catchment in Croatia Fig. 1 The Ilova River catchment in Croatia. Fig. 1 The Ilova River catchment in Croatia. 7 7 7 river mouth of the Ilova River into the Sava River. For the purpose of this paper, the third mentioned location with the river mouth of the Ilova into the Sava River (at Trebež village) is chosen because it is the actual ending of the Ilova River today. The altitude of this location is 93 m. Due to the artificial bifurcation of the catchment, the Pakra River has not been included in this research. Morfološko stanje tekućica u poriječju Ilove More precisely, as a result of river training, a part of the Pakra River waters is drained towards Veliki Strug and further to Mokro Polje and the Sava River, while the other part, as previously stated, has a confluence into the Ilova River a few kilometres upstream from the river’s mouth into the Sava River. tinsko Selo. Druga je nizvodna lokacija ušće Pakre u Ilovu, a treća je današnje ušće Ilove u Savu. Za potrebe rada uzeta je treća lokacija, odnosno ušće Ilove u Savu (kod naselja Trebež) jer je to danas stvarni završetak toka Ilove. Nadmorska je visina ove lokacije 93 metra. Rijeka Pakra i njeno pori ječje nisu uključeni u ovaj rad jer je riječ o umjet noj bifurkaciji. Naime hidrotehničkim radovima dio vode rijeke Pakre odvodnjava se prema Veli kom Strugu i dalje prema Mokrom polju i Savi, dok se drugi dio, kao što je već napomenuto, ulije va u Ilovu nekoliko kilometara prije njezina ušća u Savu. The length of the Ilova River from spring to river mouth into Stari Trebež is 93.4 km. If the length of the channelized course of Stari Trebež is included, as it is part of the Ilova River course today, the total length adds up to 100.9 km, with a catchment area (excluding the Pakra River catchment) of around 1128 km² (Plan tak, 2014). This short depiction and issues regarding the definition of the river mouth, total length and catch ment size imply the complexity and results of the water works in the Ilova River catchment. For this reason, this paper is dealing with the morphological condition of the streams in the Ilova River catchment, greatly de fined by human interventions. Duljina Ilove iznosi 93,4 km od izvora do ušća u Stari Trebež. Ako pribrojimo duljinu kanaliziranog toka Starog Trebeža, koji je da nas dio toka Ilove, dolazimo do duljine Ilove od 100,9 km, dok površina poriječja (bez poriječ ja Pakre) iznosi oko 1128 km² (Plantak, 2014). Navedeni kratki opis i problematika određivanja ušća rijeke, duljine toka i njezina poriječja im pliciraju složenost i posljedice hidrotehničkih zahvata u poriječju Ilove. Upravo stoga ovaj se rad bavi morfološkim stanjem tekućica poriječja Ilove, koje je uvelike određeno upravo ljudskim intervencijama. Geomorphologically, the entire analysed catchment is located at the south-eastern part of the Croatian Central basin and mainly encompasses the sub-geo morphological region – the Ilova basin (Bognar, 2001). HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Morfološko stanje tekućica u poriječju Ilove Belonging to the Peri-pannonian area, changes between lowland and hilly relief are characteristic to the catch ment. Only the eastern part of the catchment extends to the slopes of Papuk and Ravna Gora. The absolute slope of the Ilova River amounts to 112 m, and the absolute difference between the highest and lowest points in the catchment amount to 770.5 m. Geomorfološki se promatrano poriječje u cje lini nalazi na jugoistoku Središnje zavale, a veći nom obuhvaća subgeomorfološku regiju Zavala Ilove (Bognar, 2001). Karakteristika poriječja jest pripadnost peripanonskom prostoru, u kojem se izmjenjuju nizinski i brežuljkasto-brdski reljef, a samo na istoku poriječje obuhvaća obronke Pa puka i Ravne Gore. Apsolutni pad rijeke iznosi 112 metara, a apsolutna razlika između najviše i najniže točke na poriječju 770,5 metara. Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove Materials and methods The research of morphological condition of streams in the Ilova River catchment was carried out in three steps. In the first step, the data were collected by using the analysis of available historical and contemporary maps: Topographic map 1:25,000; Digital ortho photo images; Croatian Base Map 1:5,000 (DGU Geoportal); maps of the Second and Third Military Survey of the Habsburg Monarchy (Mapire internet Istraživanje morfološkog stanja tekućica u pori ječju Ilove provedeno je u tri koraka. U prvom ko raku podatci su prikupljeni uporabom i analizom dostupnih povijesnih i suvremenih kartografskih podloga: Topografske karte 1 : 25.000, Digitalnih ortofoto snimaka, Hrvatske osnovne karte 1 : 5.000 (Geoportal Državne geodetske uprave), karte Dru ge i Treće vojne izmjere Habsburške Monarhije 8 (internetski portal Mapire), Aerofotogrametrijskog materijala iz 1968. (Geoportal Ministarstva gradi teljstva i prostornog uređenja) te digitalnog modela reljefa Aster GDEM. portal); Aerial orthophoto from 1968 (Geoportal of the Ministry of Construction and Physical Planning) and the Aster GDEM digital elevation model. Morfološko stanje tekućica u poriječju Ilove tion) of surface water bodies1 was made according to the GIS Guidance WFD (CIS GD9, 2003), but also taking into account the existing hydrotehnical struc tures, water diversions and other significant changes in the streams (CIS GD2, 2003) collected analysing topographic maps of 1:25,000 and digital orthopho to images. A total of 48 water bodies (Fig. 2) where determined of which two are artificial water bodies (canals) and, in accordance to the WFD assessment, evaluation for them was not carried out. Twelve water bodies refer to the Ilova and Toplica Rivers. Nakon prikupljanja i obrade kartografskih i srodnih podloga te utvrđivanja vodnih tijela prove deni su identifikacija većih hidrotehničkih radova koji su učinjeni na vodotocima te preliminarno ka binetsko ocjenjivanje morfološkog stanja. Uz ocje njivanje odabirane su lokacije dionica za terensko istraživanje koje je pretežno obuhvatilo veće vodo toke. Nakon terenskog istraživanja napravljeno je završno ocjenjivanje. Following the collection and processing of carto graphic and related materials and the delineation of river water bodies, identification of major river training works was carried out along with preliminary cabinet assessment of the morphological condition. During the cabinet assessment, sites for field surveying were identified, predominantly covering larger streams. The final assessment was conducted after the field survey. The assessment of the morphological condition of water bodies was carried out according to the Meth odology for Monitoring and Assessment of Hydro morphological Features (Methodology, 2015) and the Guideline for Hydromorphological Monitoring and Assessment of Rivers in Croatia (MEANDER, 2013). Both documents have been developed as the result of a capacity-building project supporting Cro atia’s harmonisation with the legislation and moni toring practices within various EU Directives. The hydromorphological status of rivers according to the WFD and the afore-mentioned Croatian Waters’ methodology (Methodology, 2015) takes into ac count: the hydrological regime, river continuity and the morphological conditions. In this paper, only the morphological condition elements were assessed (Tab 1, elements under 3 Morphology). In addition, sites of river discontinuity were also identified. However, for a precise assessment and scoring of this element, ich thyological research would be needed. Analysis and scoring of the hydrological regime is also difficult due to lack of a continuous set of historical data and the relatively small number of hydrological stations. Morfološko stanje tekućica u poriječju Ilove The Ilova River catchment area was determined using hydrological GIS tools (ArcHydro) and the AS TER GDEM digital elevation model and corrected according to the 1:25,000 topographic map. Further more, streams were digitized from the 1:25,000 topo graphic maps from the years 1999, 2000 and 2001. In total, 534 km of streams where determined and taken for analysis, of which 100.9 km is the Ilova River, 48.5 km the largest tributary, Toplica, while the remainder refers to the streams in the catchment with a min imum drainage area of 10 km². Larger determined tributaries of the Ilova River are: Tomašica, Peratovi ca, Čavlovica, Garešnica, Bršljanica, Šovarnica, Rije ka, Rastovac and Kutinica. Determination (delinea Poriječje Ilove određeno je pomoću hidroloških GIS alata (ArcHydro) i digitalnog modela reljefa ASTER GDEM te korigirano prema Topograf skoj karti 1 : 25.000. Nakon toga digitalizirani su vodotoci s Topografske karte 1 : 25.000 iz 1999., 2000. i 2001. Ukupno je utvrđeno 534 km vodo toka uzetih za analizu, od čega je 100,9 km duljina rijeke Ilove, 48,5 km duljina najvećeg pritoka rijeke Toplice, dok se ostali dio odnosi na tekućice u po riječju čije je slijevno područje minimalno 10 km². Veći tako izdvojeni pritoci Ilove jesu: Tomašica, Peratovica, Čavlovica, Garešnica, Bršljanica, Šo varnica, Rijeka i Rastovac, Kutinica. Određivanje 9 Sl. 2. Fizičkogeografska karta poriječja Ilove (brojevi označuju određena vodna tijela) Fig. 2 Physical-geographical map of the Ilova River catchment (numbers identify water bodies). Sl. 2. Fizičkogeografska karta poriječja Ilove (brojevi označuju određena vodna tijela) Sl. 2. Fizičkogeografska karta poriječja Ilove (brojevi označuju određena vodna tijela) Fig. 2 Physical-geographical map of the Ilova River catchment (numbers identify water bodies). Fig. 2 Physical-geographical map of the Ilova River catchment (numbers identify water bodies). 9 (delineacija) površinskih vodnih tijela1 napravljeno je prema GIS vodiču ODV-a (GD9, 2003) uzi majući u obzir i postojeće hidrotehničke objekte, preusmjerivanja vode i ostale značajne promjene na vodotocima (GD2, 2003) prikupljene analizom Topografske karte 1 : 25.000 i Digitalnih ortofoto snimaka. Ukupno je određeno 48 vodnih tijela (sl. 2), od kojih su dva umjetna vodna tijela (kanali) te se za njih, sukladno ODV-u, ocjenjivanje ne pro vodi. Dvanaest vodnih tijela odnosi se na Ilovu i Toplicu. HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) 1 Vodno tijelo (površinsko) prema dokumentima ODV-a (ODV, čl. 2, st. 10; GD2) predstavlja jasno odvojenu/određenu karakterističnu dionicu površin ske vode kao što su jezero, akumulacija, potok, rijeka ili kanal; dio jezera, potoka, rijeke ili kanala; prijelazne vode ili pojas priobalne vode (preveli autori iz izvornika na engleskom jeziku). 1 Body of surface water means a discrete and significant element of surface water such as a lake, a reservoir, a stream, river or canal, part of a stream, river or canal, a transitional water or a stretch of coastal water (ODV, 2000). ater means a discrete and significant element of surface water such as a lake, a reservoir, a stream, river or canal, part of a stream, river onal water or a stretch of coastal water (ODV, 2000). Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove Morfološko stanje tekućica u poriječju Ilove Ocjenjivanje morfološkog stanja vodotoka provedeno je prema dokumentima Metodologi ja monitoringa i ocjenjivanja hidromorfoloških pokazatelja (Metodologija, 2015) i Vodič za hi dromorfološki monitoring i ocjenu stanja rijeka u Hrvatskoj (MEANDER, 2013) koji su nastali u sklopu projekta potpore Hrvatskoj za ispunja vanje zahtjeva Europske unije odnosno smjernica iz Okvirne direktive Europske Unije o vodama. Hidromorfološko stanje tekućica prema ODV-u i navedenoj metodologiji Hrvatskih voda (Meto dologija, 2015) sastoji se od hidrološkog režima, uzdužne povezanosti i morfologije. U ovome radu istraženi su i ocjenjeni samo elementi morfologije (tab. 1, elementi pod 3, Morfologija). Utvrđene su i lokacije prekida uzdužne povezanosti, ali je za kvalitetnije vrjednovanje i bodovanje toga ele menta potrebno provesti ihtiološka istraživanja. Analiza i vrjednovanje hidrološkog režima ote žani su nedostatkom dovoljno dugih nizova po dataka i relativno rijetkom mrežom hidroloških stanica. 10 Bodovanje elemenata i konačna ocjena morfo loškog stanja provedeni su na temelju kvantitativ nih podataka. Bodovna kategorija s kvantitativnim podacima jest peterostupanjska ljestvica (1 = naj manje odstupanje od prirodnog stanja, 5 = najveće odstupanje od prirodnog stanja) osim za elemente 3. 2. 2., 3. 3. 1., 3. 3. 2. i 3. 4., gdje je uporabljena trostupanjska ljestvica (1 = najmanje odstupanje od prirodnog stanja, 3 = najveće odstupanje od prirod The scoring of elements and final score of the morphological condition is based on quantitative data. The score category with quantitative data consists of a five-point scale (1 = lowest degree of modification, 5 = highest degree of modification), except for the elements 3.2.2, 3.3.1, 3.3.2 and 3.4 where the qualitative data of a three-point scale (1 = lowest degree of modification, 5 = highest de gree of modification) was used. Scoring for those 11 Izvor: Metodologija monitoringa i ocjenjivanja hidromorfoloških pokazatelja (Hrvatske vode, 2015) Source: Methodology for Monitoring and Assessment of Hydromorphological Features (Hrvatske vode, 2015) Tab. 1. Elementi ocjene hidromorfološkog stanja Tab. 1 Features used for determining the hydromorphological modifications of rivers 1. Hidrologija (hidrološki režim) / Hydrology 1. 1. Učinci umjetnih građevina u koritu unutar dionice / Impacts of artificial in-channel structures within the reach 1. 2. Učinci promjena širom sliva na karakter prirodnog toka / Effects of catchment-wide modifications on natural flow character 1. 3. Učinci promjene u dnevnom protoku / Effects of daily flow alteration 2. Uzdužna povezanost / Longitudinal continuity 2. 1. Uzdužna povezanost pod utjecajem umjetnih građevina / Longitudinal connectivity affected by artificial structures 3. Morfološko stanje tekućica u poriječju Ilove Vegetacija i organski ostaci u koritu / Channel vegetation and organic debris 3. 3. 1. Uklanjanje vodene vegetacije / Aquatic vegetation management 3. 3. 2. Količina drvenih ostataka, ako se očekuju / Extent of woody debris if expected 3. 4. Karakter erozije/taloženja / Erosion/deposition character 3. 5. Struktura obale i promjene na obali / Bank structure and modifications 3. 6. Vrsta/struktura vegetacije na obalama i okolnom zemljištu / Vegetation type/structure on banks and adjacent land 3. 7. Korištenje zemljišta i s time povezana obilježja / Land-use and associated features 3. 8. Interakcija korita i poplavnog područja / Channel-floodplain interactions 3. 8. 1. Stupanj lateralne povezanosti rijeke i poplavnog područja / Degree of lateral connec-tivity of river and floodplain 3. 8. 2. Stupanj lateralnoga kretanja riječnoga korita / Degree of lateral movement of river channel 11 Izvor: Metodologija monitoringa i ocjenjivanja hidromorfoloških pokazatelja (Hrvatske vode, 2015) Source: Methodology for Monitoring and Assessment of Hydromorphological Features (Hrvatske vode, 2015) Tab. 1. Elementi ocjene hidromorfološkog stanja Tab. 1 Features used for determining the hydromorphological modifications of rivers 1. Hidrologija (hidrološki režim) / Hydrology 1. 1. Učinci umjetnih građevina u koritu unutar dionice / Impacts of artificial in-channel structures within the reach 1. 2. Učinci promjena širom sliva na karakter prirodnog toka / Effects of catchment-wide modifications on natural flow character 1. 3. Učinci promjene u dnevnom protoku / Effects of daily flow alteration 2. Uzdužna povezanost / Longitudinal continuity 2. 1. Uzdužna povezanost pod utjecajem umjetnih građevina / Longitudinal connectivity affected by artificial structures 3. Morfologija / Morphology 3. 1. Geometrija korita / Channel geometry 3. 1. 1. Tlocrtni oblik / Planform 3. 1. 2. Presjek korita (uzdužni i poprečni presjek) / Channel section 3. 2. Podloge / Substrates 3. 2 .1. Količina umjetnih tvrdih materijala / Extent of artificial material 3. 2. 2. Mješavina "prirodnih" podloga ili izmijenjena značajka / ‘Natural’ substrate mix or character altered 3. 3. Vegetacija i organski ostaci u koritu / Channel vegetation and organic debris 3. 3. 1. Uklanjanje vodene vegetacije / Aquatic vegetation management 3. 3. 2. Količina drvenih ostataka, ako se očekuju / Extent of woody debris if expected 3. 4. Karakter erozije/taloženja / Erosion/deposition character 3. 5. Struktura obale i promjene na obali / Bank structure and modifications 3. 6. Vrsta/struktura vegetacije na obalama i okolnom zemljištu / Vegetation type/structure on banks and adjacent land 3. 7. Morfološko stanje tekućica u poriječju Ilove Morfologija / Morphology 3. 1. Geometrija korita / Channel geometry 3. 1. 1. Tlocrtni oblik / Planform 3. 1. 2. Presjek korita (uzdužni i poprečni presjek) / Channel section 3. 2. Podloge / Substrates 3. 2 .1. Količina umjetnih tvrdih materijala / Extent of artificial material 3. 2. 2. Mješavina "prirodnih" podloga ili izmijenjena značajka / ‘Natural’ substrate mix or character altered 3. 3. Vegetacija i organski ostaci u koritu / Channel vegetation and organic debris 3. 3. 1. Uklanjanje vodene vegetacije / Aquatic vegetation management 3. 3. 2. Količina drvenih ostataka, ako se očekuju / Extent of woody debris if expected 3. 4. Karakter erozije/taloženja / Erosion/deposition character 3. 5. Struktura obale i promjene na obali / Bank structure and modifications 3. 6. Vrsta/struktura vegetacije na obalama i okolnom zemljištu / Vegetation type/structure on banks and adjacent land 3. 7. Korištenje zemljišta i s time povezana obilježja / Land-use and associated features 3. 8. Interakcija korita i poplavnog područja / Channel-floodplain interactions 3. 8. 1. Stupanj lateralne povezanosti rijeke i poplavnog područja / Degree of lateral connec-tivity of river and floodplain 3. 8. 2. Stupanj lateralnoga kretanja riječnoga korita / Degree of lateral movement of river channel Tab. 1. Elementi ocjene hidromorfološkog stanja Tab. 1 Features used for determining the hydromorphological modifications of rivers Izvor: Metodologija monitoringa i ocjenjivanja hidromorfoloških pokazatelja (Hrvatske vode, 2015) Source: Methodology for Monitoring and Assessment of Hydromorphological Features (Hrvatske vode, 2015) Tab. 1. Elementi ocjene hidromorfološkog stanja Tab. 1 Features used for determining the hydromorphological modifications of rivers 1. Hidrologija (hidrološki režim) / Hydrology 1. 1. Učinci umjetnih građevina u koritu unutar dionice / Impacts of artificial in-channel structures within the reach 1. 2. Učinci promjena širom sliva na karakter prirodnog toka / Effects of catchment-wide modifications on natural flow character 1. 3. Učinci promjene u dnevnom protoku / Effects of daily flow alteration 2. Uzdužna povezanost / Longitudinal continuity 2. 1. Uzdužna povezanost pod utjecajem umjetnih građevina / Longitudinal connectivity affected by artificial structures 3. Morfologija / Morphology 3. 1. Geometrija korita / Channel geometry 3. 1. 1. Tlocrtni oblik / Planform 3. 1. 2. Presjek korita (uzdužni i poprečni presjek) / Channel section 3. 2. Podloge / Substrates 3. 2 .1. Količina umjetnih tvrdih materijala / Extent of artificial material 3. 2. 2. Mješavina "prirodnih" podloga ili izmijenjena značajka / ‘Natural’ substrate mix or character altered 3. 3. Hidrotehnički radovi u poriječju Ilove An outline is given of the development of surface hy drotechnical works in the Ilova River catchment through the construction of larger fish farms, river training, con struction of embankments, removal of vegetation, etc. These water works led to the present day morphological status of streams in the analysed catchment. U nastavku se ukratko prikazuje razvoj antro pogenih utjecaja u poriječju Ilove kroz izgradnju većih ribnjaka, kanaliziranje vodotoka, izgradnju nasipa, uklanjanje vegetacije i sl. Ti su radovi do veli do današnjega morfološkog stanja rijeka u analiziranom poriječju. The construction of the Končanica fish farm around the year 1900 can be considered as the first significant hy drotechnical facility in the Ilova River catchment. In the beginning, the human impact was manifested through soil excavation and building of embankments and weirs to divert water to the fish ponds. As the ponds started to cover more area, it was necessary to relocate the entire river channel, primarily the Ilova River and the Đurđička and Crni potok (Crnaja) streams. The last fish pond was built in 1983 when the fish farm covered an area of 1,474 ha. The production surface area of the fish farm today is about 943 ha (Ribnjačarstvo Končanica, 2015). The Pol jana fish farm was founded in 1902. Until 1948, when it became the property of the FR of Croatia, 842 ha of fishponds had been built. In the last 65 years, fish farms have continued to build up to the present 1,271 ha of fishponds (Ribnjačarstvo Poljana, 2014). The Garešnica fish farm (Agrokombinat), Riba d.d. today, was founded in 1971. Fishponds are spread along the right bank of the Ilova River from the town of Garešnica to Veliko Vukov je village. The surface area of the fishponds is 561 ha (Fi nag, 2015). Due to building of the Poljana and Garešnica fish farms, the river channel of the Ilova River was relo cated and channelized from Garešnica to Veliko Vukovje in a total length of about 11 km. Fieldwork examination showed that waters from this channel are diverted and used for fish farming and that the channel is dry most of Prvim značajnijim hidrotehničkim radovima u poriječju Ilove možemo smatrati izgradnju ribnja ka Končanica oko 1900. U početku se antropoge ni utjecaj očitovao kroz iskapanje zemlje i gradnju nasipa te stvaranje ustava za preusmjerivanje vode prema ribnjacima. HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Morfološko stanje tekućica u poriječju Ilove Korištenje zemljišta i s time povezana obilježja / Land-use and associated features 3. 8. Interakcija korita i poplavnog područja / Channel-floodplain interactions 3. 8. 1. Stupanj lateralne povezanosti rijeke i poplavnog područja / Degree of lateral connec-tivity of river and floodplain 3. 8. 2. Stupanj lateralnoga kretanja riječnoga korita / Degree of lateral movement of river channel 1. Hidrologija (hidrološki režim) / Hydrology 11 nog stanja). Ocjenjivanje elemenata koji imaju tro stupanjsku ljestvicu otežano je subjektivnošću po kazatelja koje treba analizirati. Ti su elementi pod ložni subjektivnoj procjeni istraživača i poznavanju stanja na terenu jer ne postoji dovoljno konkretnih i lako mjerljivih kvantitativnih podataka te je teško odrediti koliko su određeni elementi izmijenjeni u odnosu na prirodno stanje. Konačna ocjena morfo loškog stanja za svako vodno tijelo (dionicu) srednja je vrijednost dvanaest ocijenjenih elemenata. elements with the three-point scale was somewhat difficult due to subjectivity. Lack of concrete and measurable data makes those elements highly de pendent on the researcher’s experience and field work knowledge. In fieldwork, it is hard to esti mate the level of change between the natural and present state. The final score for each water body (river reach) is the mean value of twelve assessed elements used for the evaluation of the morpho logical condition. Morfološko stanje tekućica u poriječju Ilove Uz hidrotehničke radove potrebne za izgradnju ribnjakâ na poriječju Ilove kanaliziran je velik dio tokova rijeka i potoka. Prvi veći hidrotehnički ra dovi pokrenuti su početkom šezdesetih godina 20. stoljeća. Analizom aerofotogrametrijskih snimaka iz 1968. utvrđeno je da Ilova, osim kratkih dionica uz ribnjake, do toga vremena nije bila značajnije kanalizirana te je tekla prirodnim koritom. Ma nji nasipi bili su izgrađeni na dionici od ribnja ka Končanica do Garešnice i dionici od naselja Zbjegovača do današnje Autoceste A3. Potpuno kanaliziran bio je dio toka Ilove od ušća Dišnice (Brinjani) do naselja Zbjegovača u ukupnoj du ljini od 2,4 km. S iste snimke utvrđeno je da je tada u tijeku bilo kanaliziranje rijeke Toplice od mosta između naselja Brestovačka Brda i Pehovac do starog mlina u naselju Hrastovac u ukupnoj duljini od 12 km te na dionici kroz grad Daruvar. Također su već 1968. bili kanalizirani potoci Ga rešnica od ušća uzvodno do naselja Garešničkog Brestovca, Tomašica od ušća do naselja Herce govca i Peratovica od ušća do Gornje Rašenice, dok je Kutinica bila kanalizirana samo djelomično kroz grad Kutinu. Izrazite antropogene promjene događaju se u posljednjih petnaestak godina na kon donošenja izmjena i dopuna Državnog plana zaštite od poplava iz 1997. (Državni plan, 1997), otkada se intenzivnije grade nasipi i riječne stube (stepenice), uklanja priobalna (riparijska) vegeta cija, produbljuju korita i podižu postojeći nasipi s ciljem zaštite od poplava u poriječju Ilove. Tako je ukupno od donošenja toga dokumenta u poriječju Ilove izgrađeno ili dograđeno 148 km nasipa (Dr žavni plan, 1997). In addition to hydrotechnical works needed to build fish farms in the Ilova River catchment, many rivers and streams are channelized. The first large river training project started in the 1960s. Analysis of aeri al photos from 1968 showed that the Ilova River was not significantly channelized, except for smaller parts alongside the fish farms. Smaller embankments were built on the reach between the Končanica fish farm and the town of Garešnica, and between the Zbjegovača settlement and today’s A3 highway. A 2.4 km part of the Ilova River was completely channelized from the mouth of the Dišnica Stream (Brinjani settlement) till that of Zbjegovača. Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove Hidrotehnički radovi u poriječju Ilove Kako su ribnjaci počeli zauzi mati sve veću površinu, bilo je potrebno izmje štati čitave vodotoke, prije svega Ilovu te potoke Đurđička i Crni potok (Crnaja). Zadnji ribnjak izgrađen je 1983., a danas površina ribnjaka u pro izvodnji iznosi oko 943 ha (od ukupno 1474 ha) (Ribnjačarstvo Končanica, 2015). Ribnjačarstvo Poljana nastalo je 1902. Do 1948. izgrađeno je 842 ha ribnjaka, kada ribnjaci prelaze u vlasništvo SR Hrvatske. U tom razdoblju nastavlja se gradnja do današnjeg 1271 ha ribnjaka (Ribnjačarstvo Polja na, 2014). Ribnjačarstvo Garešnica (Agrokombi nat), danas Riba d.d., nastalo je 1971. Ribnjaci se prostiru uz desnu obalu Ilove od grada Garešnice do Velikog Vukovja. Površina ribnjaka iznosi 561 ha (Finag, 2015). Zbog izgradnje ribnjaka Polja na i Ribnjačarstva Garešnica izmješteno je korito Ilove na dionici od Garešnice do Velikog Vuko vja u duljini od oko 11 km. Terenskim izlaskom ustanovljeno je da tim koritom većinu godine ne protječe voda jer je preusmjerena za opskrbu rib njakâ. Za potrebe tih ribnjaka također se upotre 12 the year. In addition, waters from Toplica River are used for the two mentioned fish farms and only small amounts of water run in the old natural river bed. The Poljodar TIM fish farm is spread along the left bank of the Toplica River between the villages of Blagorodovac and Sokolo vac, covering an area of 91 ha (Poljodar TIM, 2014). bljava voda rijeke Toplice, koja je gotovo u cijelosti usmjerena prema njima, dok starim izvornim ko ritom protječu vrlo male količine vode. Ribnjaci Poljodar TIM-a prostiru se na površini od oko 91 ha, uz lijevu obalu rijeke Toplice između sela Bla gorodovac i Sokolovac (Poljodar TIM, 2014). HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Morfološko stanje tekućica u poriječju Ilove Same photos show that the river training works of the Toplica Rive were under way be tween the bridge connecting the Brestovačka Brda and Pehovac settlements and the old mill at the Hrastovac settlement (a 12 km long reach), as well as in the town of Daruvar. In addition, the following streams were al ready channelized in 1968: Garešnica (from the mouth upstream to the Garešnički Brestovac settlement), Tomašica (from the mouth upstream to the Hercego vac settlement), Peratovica (from the mouth upstream to the Gornja Rašenica settlement) and smaller parts of the Kutinica in the town of Kutina. Larger training works are significant for the period after the adoption of a new State Flood Protection Plan (Državni plan zaštite od poplava) in 1997 (Državni plan, 1997). Fol lowing the new plan, an intensive construction of new levees, embankments and river steps occurred alongside the removal of riparian vegetation, river dredging and upgrading of levees, all with the aim of flood protection in the catchment. Following the year 1997, a total of 148 km of levees in the catchment were built or up graded (Državni plan, 1997). Od ukupno 534 km vodotokâ u poriječju Ilo ve kanalizirano je oko 250 km. Od 250 km ka naliziranih vodotoka na oko 109 km uklonjena je priobalna drvenasta vegetacija (sl. 3). Kanalizirane dionice nalaze se u većim naseljima i ribnjacima koji su u središnjem dijelu promatranog poriječja i oko njih, odnosno u srednjem dijelu toka Ilove From the total of 534 km of watercourses in the Ilova River catchment, 250 km have been channel ized. Out of those 250 km, bank (riparian) vege tation has been removed on around 109 km (Fig. 3). Channelized reaches are mainly in and around bigger settlements and fish farms in the central part of the catchment, i.e. the middle course of the Ilo 13 i glavnih pritoka Toplice, Garešnice i Šovarnice. Usto, znatno su kanalizirani i srednji i donji dio toka rijeke Kutine. Potrebno je naglasiti da je, na žalost, kanalizirano i dosta dionica vodotokâ izvan naselja te u područjima u kojima su naselja izgra đena na prirodno izdignutom terenu koji je zašti ćen od štetnog djelovanja velikih voda. Uz kana liziranje vodotokâ značajni su izgradnja nasipa na većim vodotocima i na ušćima manjih vodotoka te izgradnja mreže melioracijskih kanala, koji nisu ocjenjivani u ovome radu. va River and the Toplica, Garešnica and Šovarni ca tributaries. Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove Morfološko stanje tekućica u poriječju Ilove The middle and lower course of the Kutina River is also highly channelized. One has to point out that, unfortunately, a lot of stream reach es outside of settlements and in the areas where the settlements are built on a naturally elevated and thus flood protected terrain are channelized. In addition, a lot of new levees on bigger streams and confluences were built. A network of irrigation channels was also extended, but that is not evaluat ed in this research. Sl. 3. Kanalizirane dionice vodotokâ i dionice s uklonjenom priobalnom vegetacijom u poriječju Ilove Fig. 3 Channelized river reaches and reaches with removed riparian vegetation in the Ilova River catchment. Sl. 3. Kanalizirane dionice vodotokâ i dionice s uklonjenom priobalnom vegetacijom u poriječju Ilove Fig. 3 Channelized river reaches and reaches with removed riparian vegetation in the Ilova River catchment. Sl. 3. Kanalizirane dionice vodotokâ i dionice s uklonjenom priobalnom vegetacijom u poriječju Ilove River training as a rule has an array of negative effects. The removal of bank (riparian) woody vege tation decreases bank stability and increases erosion (Prosser et al., 2001). An example of such negative effects in the Ilova River catchment is the river reach from the outlet canal of the Končanica fish farm to the bridge on the Ilova River between Ismovac and Hidrotehnički radovi u pravilu imaju niz ne gativnih posljedica. Uklanjanjem priobalne (ripa rijske) drvenaste vegetacije destabiliziraju se oba le i povećava njihova erozija (Prosser i dr., 2001). Primjer takvih negativnih procesa u poriječju Ilove jest dionica od ispusnoga kanala ribnjaka Konča nica do mosta na Ilovi između naselja Imsovac i 14 Ilovski Klokočevac (Fig. 4). Following the hydrotech nical works and removal of vegetation, the river bank eroded and collapsed. Removal of bank (riparian) vegetation also affects the river water temperature, especially during summer months (Pusey and Arth ington, 2003; Sabater et al., 2012; Fig. 5). Ilova has a Panonnian rain regime with a minimum discharge in July and August (Čanjevac, 2013) and a combina tion of minimal discharge and direct exposure to so lar radiation endangers the river ecosystem (Bonacci, 2003). Ilovski Klokočevac (sl. 4). Na tom je mjestu nakon hidrotehničkih radova došlo do urušavanja i ero zije desne obale, na kojoj je uklonjena vegetacija. Uklanjanje priobalne vegetacije izrazito utječe i na temperaturu vode tijekom ljetnih mjeseci (Pusey i Arthington, 2003; Sabater i dr., 2012) (sl. 5). Sl. 4. Erozija obale Ilove kod na selja Ilovski Klokočevac uslijed uklanjanja vegetacije na lijevoj obali (desni dio fotografije) Morfološko stanje tekućica u poriječju Ilove Ilova ima Panonski kišni režim, s minimumima proto ka upravo u srpnju i kolovozu (Čanjevac, 2013), te kombinacija minimalnih protoka i izravne izlože nosti Sunčevu zračenju ugrožava riječni ekosustav (Bonacci, 2003). Ilovski Klokočevac (sl. 4). Na tom je mjestu nakon hidrotehničkih radova došlo do urušavanja i ero zije desne obale, na kojoj je uklonjena vegetacija. Uklanjanje priobalne vegetacije izrazito utječe i na temperaturu vode tijekom ljetnih mjeseci (Pusey i Arthington, 2003; Sabater i dr., 2012) (sl. 5). Ilova ima Panonski kišni režim, s minimumima proto ka upravo u srpnju i kolovozu (Čanjevac, 2013), te kombinacija minimalnih protoka i izravne izlože nosti Sunčevu zračenju ugrožava riječni ekosustav (Bonacci, 2003). Sl. 6. Stari sifon iz ribnjaka Končanica i današnja razina rijeke Ilove HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Rezultati i rasprava A total length of 534 km of watercourses in the Ilova River catchment was investigated. For the purpose of morphological evaluation, rivers were divided into reaches (Tab. 2) on the basis of length, tributaries, river bed drop or hydrotechnical works. Istraživanjem je analizirano 534 km vodotoka u poriječju Ilove. Određeni dulji vodotoci koji zbog pritoka, nagiba korita, opsežnih hidrotehničkih ra dova mijenjaju karakter podijeljeni su na određene dionice (tab. 2). Rezultati ocjenjivanja 534 km vodotokâ odno sno 48 vodnih tijela, od kojih su dva umjetna uku pne duljine od 10,4 km, prema pojedinim elemen tima te završne ocjene navedeni su u tab. 3 i prika zani na sl. 7. Ukupno je 4,3 km vodotokâ (dionica Toplica 3) dobilo završnu ocjenu vrlo loše morfološko stanje, 58,2 km, odnosno sedam dionica, ocjenu loše morfološko stanje, 143,5 km vodotoka, odnosno tri naest dionica, umjereno promijenjeno morfološko sta nje, 278,7 km, odnosno devetnaest dionica, dobro morfološko stanje i 38,4 km, odnosno šest dionica, vrlo dobro morfološko stanje. Ukupno 38,5 % duljine određenih vodotokâ u poriječju Ilove nema zadovo ljavajuću ocjenu prema Okvirnoj direktivi Europske Unije o vodama jer je dobilo ocjenu umjereno pro mijenjeno, loše ili vrlo loše morfološko stanje. Vodotoci u blizini većih naselja dobili su, očekivano, najlošije, odnosno nezadovoljavajuće ocjene. Usto su dionice koje su neposredno povezane s ribnjacima također dobile nezadovoljavajuće ocjene. Područja su to naj većih vodotoka (Ilova, Toplica) s najvećim i dugo trajnim antropogenim utjecajima. Zabrinjavajuće je da su nezadovoljavajuće ocjene dobile i mnoge di onice izvan naselja te su problematične svrha, učin kovitost i posljedice hidrotehničkih radova provo đenih u tim područjima. Zadovoljavajuće, odnosno ocjene dobro i vrlo dobro morfološko stanje dobile su uglavnom dionice iznad 200 m nadmorske visine, ponajprije izvorišne manje tekućice. The results of morphological assessment and clas sification of 534 km of watercourses, i.e. 48 water bodies (reaches) in the Ilova River catchment are presented in Tab. 3 and on Fig. 7. In total, 4.3 km of watercourses (Toplica 3 reach) received a final score of Bad morphological condition; 58.2 km of water courses, i.e. seven water bodies - Poor; 143.5 km, i.e. thirteen water bodies - Moderate; 278.7 km, i.e. nineteen water bodies - Good and 38.4 km, i.e. six water bodies received a final score of Very Good. In total, 38.5% of watercourse length in the Ilova River catchment, i.e. Morphological State of Rivers in the Ilova River Catchment Morfološko stanje tekućica u poriječju Ilove This can cause a drop in the water table, more frequent drought occurrence and increased erosion in the whole area. On channelized reaches, natural morphological characteristics, i.e. different habitats including deeper and shallower parts, rap ids, shoals and similar are missing, which negatively affects river ecology. Ilova River. This can cause a drop in the water table, more frequent drought occurrence and increased erosion in the whole area. On channelized reaches, natural morphological characteristics, i.e. different habitats including deeper and shallower parts, rap ids, shoals and similar are missing, which negatively affects river ecology. Morfološko stanje tekućica u poriječju Ilove Morfološko stanje tekućica u poriječju Ilove Sl. 4. Erozija obale Ilove kod na selja Ilovski Klokočevac uslijed uklanjanja vegetacije na lijevoj obali (desni dio fotografije) In addition to channelization and removal of woody riparian vegetation, dredging and channel deepening is present on certain reaches, which af fects the morphology of the watercourse. Channel deepening, especially if riparian woody vegetation is removed, leads to a greater flow velocity, bank erosion and lowering of the water table in the surrounding area (Svetličić, 1987). It also causes a fall in the local erosion base level, affecting stream network and increasing the channel erosion. An example of that process is shown on Fig. 6 where one can compare the Ilova river bed level after the regulations 50 years ago and recent regulations. The old outlet (siphon) from the Konačnica fish farm is around 1.4 m above the present river bed of the Uz kanaliziranje i uklanjanje priobalne drvenaste ve getacije prisutno je produbljivanje korita na nekim dioni cama, što se odražava na morfologiju vodotoka. Naime na kanaliziranim dionicama, posebice ako je uklonjena prio balna drvenasta vegetacija, dolazi do bržeg otjecanja vode. Istovremeno se javljaju veće brzine otjecanja, što dovodi do daljnjih erozija i produbljivanja korita. Produbljivanje ko rita dovodi i do spuštanja razine podzemnih voda u okol nom području (Svetličić, 1987). Produbljivanje u glavnom vodotoku utječe na smanjenje lokalne erozijske baze kojoj se moraju prilagoditi pritoci te je, bez obzira na to jesu li pritoci u prirodnom stanju ili su hidrotehnički regulirani, uočljiva pridnena erozija u koritu. Primjer produbljivanja riječnoga korita jest Ilova kod naselja Končanica (sl. 6), gdje se može usporediti korito Ilove nakon regulacija prije 15 HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Sl. 5 Ilov veg Sok Fig. lize bet Tom ripa rem Sl. 6. Stari sifon iz ribnjaka Končanica i današnja razina rijeke Ilove Fig. 6 An outlet (siphon) from the Končanica fish farm and the Ilova River water level. 16 pedesetak godina i nakon nedavnih regulacija. Mjerenjem razine starog sifona iz Ribnjaka Končanica i današnjeg dna korita utvrđeno je da je na toj lokaciji korito produbljeno oko 1,4 metra, što može utjecati na pad razine podzemnih voda, učestaliju pojavu suša te pojačanu eroziju. Na kanali ziranim dijelovima također nedostaje prirodnih morfološ kih karakteristika unutar korita, odnosno raznolikosti sta ništa koje uključuje dublje i pliće dijelove, brzice, prudove i sl., što negativno utječe na riječni ekosustav. Ilova River. Rezultati i rasprava twenty-one water bodies, do not meet the objectives of the EU Water Framework Directive (WFD) as they were classified as Moderate, Poor or Bad with respect to morphological condition. Rivers in the vicinity of larger settlements expectedly re ceived the worst, respectively failing scores. In ad dition, the assessed reaches directly associated with the fish farms also received failing scores. These areas are around the largest rivers (Ilova, Toplica), where the most extensive and long-term human impact is present. The fact that many assessed river reaches far beyond settlements also have a failing morphologi cal condition is worrying and calls into question the purpose, effectiveness and implications of the water works in these remote areas. The reaches assessed with Good and Very Good morphological condi tions are generally located over 200 m above sea level and are mainly smaller streams near the spring area. HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Rezultati i rasprava 17 p ( ) Naziv dionice / Name of reach Opis / Description Ilova 1 Od današnjeg ušća Ilove u Savu do ušća Kutinice / From current mouth of the Ilova River into the Sava River to the mouth of the Kutinica River Ilova 2 Nastavlja se na Ilovu 1 do ispusta iz ribnjaka Poljana / From the ending of Ilova 1 reach to the Poljana fish farm outlet Ilova 3 Od Ilove 2 kroz ribnjake Poljana do brane za usmjerivanje vode prema ribnjacima kod Garešnice / From Ilova 2 flowing through Poljana fish farm to the dam for Garešnica fish farm Ilova 4 Od Ilove 3 do ispusta vode iz ribnjaka Končanica kod naselja Stražanac / From Ilova 3 to the water outlet from Končanica fish farm at Stražanac Ilova 5 Od Ilove 4 do ušća potoka Peratovica / From Ilova 4 to the mouth of Peratovica stream Ilova 6 Od Ilove 5 do ušća Rijeke / From Ilova 5 to the mouth of Rijeka stream Ilova 7 Od Ilove 6 do izvora / From Ilova 6 to the spring Toplica 1 Staro korito od ušća u Ilovu uz ribnjake do spoja s Novom Toplicom / Old river bed from the mouth into Ilova River, passing the fish farms till the confluence with Nova Toplica Toplica 2 Od Toplice 1 do ušća potoka Dabrovac / From Toplica 1 to the mouth of Dabrovac stream Toplica 3 Od Toplice 2 kroz Daruvar do sportskog ribnjaka Daruvar / From Toplica 2, through the town of Daruvar till the Daruvar sport fish pond Toplica 4 Od Toplice 3 do izvora Velike Toplice / From Toplica 3 to the Velika Toplica spring Kutinica 1 Od ušća u Ilovu do naselja Ciglenica / From the river mouth to the Ciglenica settlement Kutinica 2 Od Kutinice 1 do izvora / From Kutinica 1 to the spring Garešnica 1 Od ušća u Ilovu do naselja Veliki Pašijan / From the river mouth to the Veliki Pašijan settlement Garešnica 2 Od Garešnice 1 do jezera u naselju Podgarić / From Garešnica 1 to the lake in Podgarić settlement Peratovica 1 Od ušća u Ilovu do ušća potoka Dapčevica / From the river mouth to the mouth of Dapčevica stream Peratovica 2 Od Peratovice 1 do izvora / From Peratovica 1 to the spring Čavlovica 1 Od ušća do ceste između naselja Dežanovac i Kaštel / From the river mouth to the road between Dežanovac and Kaštel settlements Čavlovica 2 Od Čavlovice 1 do izvora Crne rijeke / From Čavlovica 1 to the Crna Rijeka spring Naziv dionice / Name of reach Opis / Description Ilova 1 Od današnjeg ušća Ilove u Savu do ušća Kutinice / From current mouth of the Ilova River into the Sava River to the mouth of the Kutinica River Ilova 2 Nastavlja se na Ilovu 1 do ispusta iz ribnjaka Poljana / From the ending of Ilova 1 reach to the Poljana fish farm outlet Ilova 3 Od Ilove 2 kroz ribnjake Poljana do brane za usmjerivanje vode prema ribnjacima kod Garešnice / From Ilova 2 flowing through Poljana fish farm to the dam for Garešnica fish farm Ilova 4 Od Ilove 3 do ispusta vode iz ribnjaka Končanica kod naselja Stražanac / From Ilova 3 to the water outlet from Končanica fish farm at Stražanac Ilova 5 Od Ilove 4 do ušća potoka Peratovica / From Ilova 4 to the mouth of Peratovica stream Ilova 6 Od Ilove 5 do ušća Rijeke / From Ilova 5 to the mouth of Rijeka stream Ilova 7 Od Ilove 6 do izvora / From Ilova 6 to the spring Toplica 1 Staro korito od ušća u Ilovu uz ribnjake do spoja s Novom Toplicom / Old river bed from the mouth into Ilova River, passing the fish farms till the confluence with Nova Toplica Toplica 2 Od Toplice 1 do ušća potoka Dabrovac / From Toplica 1 to the mouth of Dabrovac stream Toplica 3 Od Toplice 2 kroz Daruvar do sportskog ribnjaka Daruvar / From Toplica 2, through the town of Daruvar till the Daruvar sport fish pond Toplica 4 Od Toplice 3 do izvora Velike Toplice / From Toplica 3 to the Velika Toplica spring Kutinica 1 Od ušća u Ilovu do naselja Ciglenica / From the river mouth to the Ciglenica settlement Kutinica 2 Od Kutinice 1 do izvora / From Kutinica 1 to the spring Garešnica 1 Od ušća u Ilovu do naselja Veliki Pašijan / From the river mouth to the Veliki Pašijan settlement Garešnica 2 Od Garešnice 1 do jezera u naselju Podgarić / From Garešnica 1 to the lake in Podgarić settlement Peratovica 1 Od ušća u Ilovu do ušća potoka Dapčevica / From the river mouth to the mouth of Dapčevica stream Peratovica 2 Od Peratovice 1 do izvora / From Peratovica 1 to the spring Čavlovica 1 Od ušća do ceste između naselja Dežanovac i Kaštel / From the river mouth to the road between Dežanovac and Kaštel settlements Čavlovica 2 Od Čavlovice 1 do izvora Crne rijeke / From Čavlovica 1 to the Crna Rijeka spring 18 M. Rezultati i rasprava Plantak, I. Čanjevac, I. Vidaković 19 Sl. 7. Ukupna morfološka ocjena vodotoka u poriječju Ilove Fig. 7 Total morphological status of rivers in the Ilova River catchment. Sl. 7. Ukupna morfološka ocjena vodotoka u poriječju Ilove Fig. 7 Total morphological status of rivers in the Ilova River catchment. Sl. 7. Ukupna morfološka ocjena vodotoka u poriječju Ilove Fig. 7 Total morphological status of rivers in the Ilova River catchment. 19 19 20 Rb. dionice / Number Dionica- ocjenjivano obilježje / Reach-Assesed element 3.1.1. 3.1.2. 3.2.1. 3.2.2. 3.3.1. 3.3.2. 3.4. 3.5. 3.6. 3.7. 3.8.1. 3.8.2. Prosječna ocjena / Average score Opisna ocjena / Status 1 Ilova 1 4 4 1 1 3 3 3 1 1 1 2 4 2,33 Dobro / Good 2 Ilova 2 3 3 1 3 1 3 3 2 2 3 2 3 2,42 Dobro / Good 3 Ilova 3 5 5 1 3 5 5 3 2 5 4 5 5 4,00 Loše / Poor 4 Ilova 4 5 5 1 3 5 5 3 2 5 4 5 5 4,00 Loše / Poor 5 Ilova 5 4 5 1 3 3 3 3 2 3 4 5 5 3,42 Umjereno / Moderate 6 Ilova 6 5 5 2 3 3 3 3 2 2 3 3 4 3,17 Umjereno / Moderate 7 Ilova7 2 2 1 1 1 1 1 1 2 3 2 2 1,58 Dobro / Good 8 Ilova – Toplica 5 5 1 3 3 3 3 2 3 3 4 5 3,33 Umjereno / Moderate 9 Toplica 1 3 3 2 3 1 1 1 1 2 3 3 3 2,17 Dobro / Good 10 Toplica 2 5 5 2 3 3 3 5 2 3 3 4 4 3,50 Loše / Poor 11 Toplica 3 5 5 3 5 3 5 5 5 5 5 5 5 4,67 Vrlo loše / Bad 12 Toplica 4 2 2 1 1 1 1 1 2 2 3 2 2 1,67 Dobro / Good 13 Kutinica 1 4 5 4 5 3 3 5 4 4 4 5 4 4,17 Loše / Poor 14 Kutinica 2 2 2 1 3 3 3 3 2 2 2 2 2 2,25 Dobro / Good 15 Lipa 4 4 1 3 3 3 3 1 4 4 3 4 3,08 Umjereno / Moderate 16 Lat. HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Rezultati i rasprava kanal Kutina 5 5 1 3 3 3 3 1 5 4 4 4 3,42 Umjereno / Moderate 17 Batinski jarak 1 2 1 1 1 1 3 1 1 3 3 3 1,75 Dobro / Good 18 Seliški potok 2 3 1 1 1 1 1 1 2 3 1 1 1,50 Dobro / Good 19 Dišnica Brinjani 3 3 1 1 1 1 1 1 3 3 2 2 1,83 Dobro / Good 20 Stupovača 4 4 1 1 1 1 1 3 2 4 4 4 2,50 Umjereno / Moderate Tab. 3. Morfološka ocjena vodotokâ prema ocjenjenim elementima Tab. Zaključak Prihvaćanjem i postupnom implementacijom Okvirne direktive Europske Unije o vodama Hr vatska je preuzela obavezu praćenja (monitoringa) i ocjene hidromorfološkog stanja vodotokâ. Ne davno su, u siječnju 2015., Hrvatske vode objavile dokument Metodologija monitoringa i ocjenjivanja hidromorfoloških pokazatelja te se na toj metodo logiji temeljilo morfološko ocjenjivanje u ovome radu. Istraživano područje, odnosno poriječje rijeke Ilove u Središnjoj Hrvatskoj veličine 1128 km2 s ukupnom duljinom riječne mreže od 534 km, podi jeljeno je na 48 vodnih tijela, koja su ocijenjena pre ma dvanaest morfoloških obilježja. Ukupno 38,5 % duljine riječne mreže u poriječju Ilove, odnosno 21 vodno tijelo, nema zadovoljavajuću ocjenu prema Okvirnoj direktivi EU-a o vodama. Riječ je o vodo tocima i dionicama u blizini većih naselja i ribnjaka. No, nažalost, nezadovoljavajuću ocjenu dobile su i mnoge dionice izvan naselja. Područja su to najve ćih i dugotrajnih antropogenih utjecaja, ali neka su od njih i nedavno hidrotehnički izmijenjena, s upit nim rezultatima i posljedicama na ukupno ekološko stanje. Zadovoljavajuću, odnosno ocjenu dobro i vrlo dobro morfološko stanje dobile su uglavnom dionice izvorišnih manjih tekućica. Za ocjenu ukupnoga hidromorfološkog stanja nedostaju detaljniji hidro loški i ihtiološki podaci o migraciji vrsta. With the acceptance and progressive implementa tion of the EU Water Framework Directive, Croatia undertook the commitment of monitoring and as sessment of the hydromorphological status of water bodies. Recently, in January 2015, Croatian Waters published a document: Methodology for monitoring and assessment of hydromorphological features based on which the assessment of the morphological condition in this research was conducted. The research area, respectively the Ilova River catchment in Central Croatia, has an area of 1128 km2. The total river network length of 534 km has been divided into 48 water bodies for which an assessment of twelve morphological features was con ducted. In total, 38.5% of watercourse length in the Ilova River catchment, respectively 21 water bodies, do not meet EU Water Framework Directive objectives. Most of these water bodies (reaches) are in the vicini ty of larger settlements and fish farms. Unfortunately, many remote river reaches were also assessed with a failing status. These rivers and river reaches have been under the most extensive and long-term human im pact. However, some have only recently been modified by water works with questionable results and outcomes on ecological status. Rezultati i rasprava 3 Morphological condition of water bodies according to assessed features 20 21 Bršljanica 2 2 1 1 1 1 1 1 2 4 3 3 1,83 Dobro / Good 22 Bršljanac 2 2 1 1 1 1 1 1 1 2 1 1 1,25 Vrlo dobro / Very good 23 Garešnica 1 5 5 2 3 3 3 3 2 4 3 5 5 3,58 Loše / Poor 24 Garešnica 2 3 3 1 3 1 3 3 1 3 2 2 2 2,25 Dobro / Good 25 Dišnica Dišnik 4 4 1 3 1 1 3 1 2 3 2 4 2,42 Dobro / Good 26 Popovača 4 4 1 3 1 1 3 1 2 3 3 3 2,42 Dobro / Good 27 Tomašica 4 4 1 3 1 3 3 1 2 4 4 4 2,83 Umjereno / Moderate 28 Matakovac 4 4 1 3 1 1 3 1 1 4 4 4 2,58 Umjereno / Moderate 29 Šovarnica 5 5 1 3 3 3 5 1 4 4 4 5 3,58 Loše / Poor 30 Peratovica 1 4 4 1 3 1 3 3 1 3 4 4 4 2,92 Umjereno / Moderate 31 Peratovica 2 2 2 1 1 1 1 3 1 2 3 1 1 1,58 Dobro / Good 32 Lončarica 1 1 1 1 1 1 1 1 1 2 1 1 1,08 Vrlo dobro / Very good 33 Rašenički potok 4 4 1 3 1 3 3 1 4 3 3 3 2,75 Umjereno / Moderate 34 Dapčevica 1 1 1 1 1 1 1 1 1 3 2 1 1,25 Vrlo dobro / Very good 35 Rastovac 1 1 1 1 1 1 1 1 1 2 2 1 1,17 Vrlo dobro / Very good 36 Kipska 2 2 1 1 1 1 1 1 1 2 2 1 1,33 Vrlo dobro / Very good 37 Jova 2 2 1 1 1 1 1 1 2 2 2 2 1,50 Dobro / Good 38 Rijeka 2 2 1 1 1 1 1 1 2 3 2 2 1,58 Dobro / Good 39 Krivaja 2 2 2 1 1 1 1 1 2 3 2 2 1,67 Dobro / Good 40 Šandrovac 1 1 1 1 1 1 1 1 1 3 1 1 1,17 Vrlo dobro / Very good 41 Dabrovac 3 3 1 3 3 3 3 1 3 3 4 4 2,83 Umjereno / Moderate 42 Čavlovica 1 4 4 1 3 3 3 3 1 2 4 4 4 3,00 Umjereno / Moderate 43 Čavlovica 2 2 2 1 1 1 1 1 1 1 3 2 2 1,50 Uobro / Good 44 Sovjak 5 5 1 3 3 3 1 1 5 5 5 5 3,50 Loše / Poor 45 Stubljarica 3 3 1 1 1 1 1 1 2 3 2 2 1,75 Dobro / Good 46 Đurđička 4 4 1 3 3 3 3 1 2 3 4 4 2,92 Umjereno / Moderate 21 HRVATSKI GEOGRAFSKI GLASNIK 78/1, 5−24 (2016.) Zaključak The reaches that meet the objec tives for morphological conditions and were assessed with Good and Very Good morphological condition are mainly the small streams near the spring area. Utvrđeno, relativno loše, morfološko stanje riječ ne mreže negativna je posljedica dugotrajnoga jed noobraznog upravljanja vodotocima u poriječju Ilo ve. Potreban je nov pristup uređenju vodotokâ koji će voditi računa o hidromorfološkim obilježjima te posljedicama morfološke degradacije na ukupni riječni ekosustav, ali i hidrološke promjene poput pojave visokih voda i snižavanja razine podzemnih voda. Ulazak u Europsku uniju potiče Hrvatsku i njezin sustav upravljanja vodnim resursima na pri hvaćanje novih paradigmi i pristupa koji će dovesti do cjelovitog i održivog upravljanja vodnim resur sima. Zbog nedovoljne objektivnosti i konkretnosti dijelova upotrijebljene metodologije te raznolikosti hidromorfoloških obilježja tekućica u Hrvatskoj nameće se i potreba detaljnijeg i boljeg određivanja i ocjenjivanja hidromorfoloških pokazatelja u Hr vatskoj. This finding of a relatively poor morphological con dition of the river network in the Ilova River catch ment is the negative outcome of a long-term one-sid ed management of rivers in the catchment. A new approach to maintenance of rivers is needed. One is needed that will take into account the hydromorpho logical features and consequences of morphological degradation for the river ecosystem as a whole. In ad dition, the influence on hydrology of the area such as occurrence of floods and groundwater level depletion should be taken into account. Accession to the EU encourages Croatia and the water resource manage ment system to accept new paradigms and approaches, which will lead to an integral and sustainable manage ment of water resources. The low precision of some parts of the methodology used and the diversity of the hydromorphological characteristics of rivers in Croatia invites researchers to improve methodology for assess ment of the hydromorphological features of Croatia. 22 M. Plantak, I. Čanjevac, I. Vidaković Barbalić, D., Kuspilić, N., 2014: Trendovi indikatora hidroloških promjena, Građe vinar 66 (7), 613-624. Čanjevac, I., Orešić, D., 2015: Contem porary Changes of Mean Annual and Seasonal River Discharges in Croatia, Hrvatski geografski glasnik 77 (1), 7-27. Plantak, M., 2014: Promjene režima otjeca nja u porječju rijeke Ilove, diplomski rad, Geografski odsjek, PMF. Prosser, I. P., Rutherfurd, I. D., Olley, J. M., Young, W. J., Wallbrink, P. J., Mo ran, C. 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Ribnjačarstvo Poljana, http://www.ribnjacarstvo-poljana.hr (10. 2. 2014.) 23 Mladen Plantak mag. geogr., doktorand, Elektroprojekt, d.d., Odjel zaštite voda, prirode i okoliša, Alexandera von Humboldta 4, 10 000 Zagreb, Hrvatska Ivan Čanjevac canjevac@geog.pmf.hr dr. sc., viši asistent, Sveučilište u Zagrebu, Prirodoslovno - matematički fakultet, Geografski odsjek, Marulićev trg 19/II, 10 000 Zagreb, Hrvatska Iva Vidaković prof. biol., doktorandica, Elektroprojekt, d.d., Odjel zaštite voda, prirode i okoliša, Alexandera von Humboldta 4, 10 000 Zagreb, Hrvatska Mladen Plantak mag. geogr., doktorand, Elektroprojekt, d.d., Odjel zaštite voda, prirode i okoliša, Alexandera von Humboldta 4, 10 000 Zagreb, Hrvatska Ivan Čanjevac canjevac@geog.pmf.hr dr. sc., viši asistent, Sveučilište u Zagrebu, Prirodoslovno - matematički fakultet, Geografski odsjek, Marulićev trg 19/II, 10 000 Zagreb, Hrvatska Iva Vidaković prof. biol., doktorandica, Elektroprojekt, d.d., Odjel zaštite voda, prirode i okoliša, Alexandera von Humboldta 4, 10 000 Zagreb, Hrvatska 24	84
https://openalex.org/W2904007649	OpenAlex	Open Science	CC-By	2,018	A new mouth brooder species of Gymnogeophagus with hypertrophied lips (Cichliformes: Cichlidae)	Andréia Turcati	English	Spoken	5,831	13,510	Neotropical Ichthyology, 16(4): e180118, 2018 DOI: 10.1590/1982-0224-20180118 Neotropical Ichthyology, 16(4): e180118, 2018 DOI: 10.1590/1982-0224-20180118 Journal homepage: www.scielo.br/ni Published online: 06 December 2018 (ISSN 1982-0224) Printed: 07 December 2018 (ISSN 1679-6225) Copyright © 2018 Sociedade Brasileira de Ictiologia Copyright © 2018 Sociedade Brasileira de Ictiologia Original article Andréia Turcati1, Wilson Sebastián Serra-Alanis2 and Luiz R. Malabarba1 A new mouth breeder species of Gymnogeophagus is described from a tributary of the río Uruguay. It is distinguished from most species of the genus by the presence of hypertrophied lips, and from G. labiatus and G. pseudolabiatus by the color pattern. The presence of successive allopatric species of the Gymnogeophagus gymnogenys clade inhabiting the tributaries of the río Uruguay is discussed. Keywords: Distribution, Endemism, Gymnogeophagus gymnogenys clade, New species, Río Uruguay. Una nueva especie incubadora bucal de Gymnogeophagus es descripta de un tributario del Río Uruguay. Se distingue de la mayoría de las especies del género por la presencia de labios hipertrofiados, y de G. labiatus y G. pseudolabiatus por su patrón de coloración. Se discute la presencia de sucesivas especies alopátricas del clado Gymnogeophagus gymnogenys ha- bitando los tributarios del Río Uruguay. Palabras Clave: Distribución, Endemismo, Clado Gymnogeophagus gymnogenys, Especie nueva, Río Uruguay. 1Programa de Pós-Graduação em Biologia Animal, Departamento de Zoologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9500, 91501-970 Porto Alegre, RS, Brazil, (AT) bioturcati@gmail.com, (LRM) malabarb@ufrgs.br, https://orcid.org/0000- 0002 9607 3735 (corresponding author) 1Programa de Pós-Graduação em Biologia Animal, Departamento de Zoologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9500, 91501-970 Porto Alegre, RS, Brazil, (AT) bioturcati@gmail.com, (LRM) malabarb@ufrgs.br, https://orcid.org/0000- 0002-9607-3735 (corresponding author) 2Sección Ictiología, Dpto. de Zoología, Museo Nacional de Historia Natural, CC 399, Montevideo, Uruguay. Centro Universitario Regional del Este (CURE), Sede Rocha, Ruta 15 y Ruta 9, Rocha, Uruguay, serraelbicho@gmail.com 1Programa de Pós-Graduação em Biologia Animal, Departamento de Zoologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9500, 91501-970 Porto Alegre, RS, Brazil, (AT) bioturcati@gmail.com, (LRM) malabarb@ufrgs.br, https://orcid.org/0000- 0002-9607-3735 (corresponding author) 2Sección Ictiología Dpto de Zoología Museo Nacional de Historia Natural CC 399 Montevideo Uruguay Centro Universitario Regional A new mouth brooder species of Gymnogeophagus with hypertrophied lips (Cichliformes: Cichlidae) ndréia Turcati1, Wilson Sebastián Serra-Alanis2 and Luiz R. Malabarba Introduction Two major clades of Gymnogeophagus are recognized: one containing substrate e180118[1] Neotropical Ichthyology, 16(4): e180118, 2018 2 New Gymnogeophagus from lower río Uruguay Fig. 1. Anterior portion of dorsal-fin skeleton of Gymno- geophagus peliochelynion (anterior towards left), UFRGS 8076, paratype, 74.2 mm SL. Arrow indicates the anterior spine in the first dorsal pterygiophore. Note the lack of su- praneurals between supraoccipital and first dorsal pterygio phore. 2015, and G. tiraparae González-Bergonzoni, Loureiro & Oviedo, 2009. In this paper we describe a new species of the G. gymnogenys clade from a tributary of the lower río Uruguay, the río Arapey Grande. Material and Methods Examined material of Gymnogeophagus belong to the fish collections of the Museu de Ciências e Tecnologia, Pon- tifícia Universidade Católica do Rio Grande do Sul, Porto Alegre (MCP), Museo Nacional de Historia Natural, Mon- tevideo (MHNM), Universidade Federal do Rio Grande do Sul, Porto Alegre (UFRGS), and Facultad de Ciencias de la Universidad de la República, Montevideo (ZVC-P). Additional comparisons were done using data or specimens listed by Reis, Malabarba (1988), González-Bergonzoni et al. (2009), Malabarba et al. (2015) and Loureiro et al. (2016). Counts and measurements were taken according to Malabarba et al. (2015) and asterisks represents counts from the holotype. Specimens listed as non-types are referred for geographical distribution record, and were not counted or measured for species description. The localities of collection of specimens listed under Comparative material session are also given for distributional purposes. Fig. 1. Anterior portion of dorsal-fin skeleton of Gymno- geophagus peliochelynion (anterior towards left), UFRGS 8076, paratype, 74.2 mm SL. Arrow indicates the anterior spine in the first dorsal pterygiophore. Note the lack of su- praneurals between supraoccipital and first dorsal pterygio phore. 13210, 1 male 76.3 mm SL, río Arapey, Colonia Lavalleja, Paso Elías, 31°02’50.94”S 57°00’46.04”W, 17 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart, S. Paullier. Gymnogeophagus peliochelynion, new species Non-types: Uruguay, Departamento de Salto. Río Arapey drainage: ZVC-P 7016, 16, 43.3-94.1 mm SL, Río Arapey Chico, Ruta 4, 31°02’07.44”S 56°53’50.21”W, 22 Nov 2005, F. Teixeira, A. D’Anatro, I. González, S. Oviedo & M. Lou- reiro. ZVC-P 12656, 3, 48.9-59.0 mm SL, Arroyo Sopas, Paso del Cementerio, 31°23’34.21”S 56°42’31.06”W, 13 Aug 2013, M. Loureiro, J. Bessonart, S. Serra & L. Montes de Oca. ZVC-P 12715, 4, 44.6-61.6 mm SL, Arroyo Sauce del Arapey, 31°19’27.25”S 56°37’24.55”W, 11 Aug 2013, M. Loureiro, J. Bessonart, S. Serra & L. Montes de Oca. ZVC-P 13013, 1, 51.7 mm SL, Arroyo Mataperros, 30°58’47.46”S 56°23’15.60”W, 15 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P 13043, 4, 43.9-59.4 mm SL, Arroyo Arerunguá, Paso del Potrero, 31°27’07.02”S 56°50’37.01”W, 18 Dec 2014, M. Loureiro, A. Duarte & S. Serra. ZVC-P 13057, 2, 65.3-90.2 mm SL, Arroyo Sopas, Paso del Cementerio, 31°23’34.21”S 56°42’31.06”W, 18 Dec 2014, M. Loureiro, A. Duarte & S. Serra. ZVC-P 13079, 3, 57.6-77.1 mm SL, Arroyo Mataojo, Pueblo Fernandez, 31°10’37.01”S 56°21’56.54”W, 14 Dec 2014, M. Lourei- ro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P 13118, 6, 56.3-77.2 mm SL, Río Arapey, Picada Sarandí, 31°03’52.47”S 56°22’44.84”W, 15 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P 13139, 2, 52.7-57.7 mm SL, Arroyo Mataojo Grande, Paso de la Her- rería, 31°11’45.40”S 56°36’11.65”W, 14 Dec 2014, M. Lou- reiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P 13178, 3, 51.2-55.9 mm SL, Río Arapey, Colonia Lavalleja, Paso Elías, 31°02’50.94”S 57°00’46.04”W, 16 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. urn:lsid:zoobank.org:act:ECB36EC2-2EE7-438B-A9B4- F28FD3899154 Introduction breeder species, whose females deposit adhesive eggs on a surface and one or both parents guard and tend them, and another that includes mouth breeder species, where one or both parents brood orally the eggs and young (Reis, Mala- barba, 1988; Wimberger et al., 1998). The substrate breeder clade has been treated as the G. rhabdotus clade, G. rhabdo- tus species group or G. rhabdotus group (Malabarba et al., 2010; Loureiro et al., 2016; Říčan et al., 2018, respectively) and includes G. rhabdotus (Hensel, 1870), G. che Casciotta, Casciotta, Gómez & Toresanni, 2000, G. meridionalis Reis & Malabarba, 1988, G. setequedas Reis, Malabarba & Pa- vanelli, 1992, G. taroba Casciotta, Almirón, Piálek & Říčan, 2017, and G. terrapurpura Loureiro, Zarucki, Malabarba & González-Bergonzoni, 2016. The mouth breeder clade has been treated as the G. gymnogenys clade and G. gym- nogenys group (Malabarba et al., 2010; Říčan et al., 2018, respectively), and includes G. gymnogenys (Hensel, 1870), G. australis (Eigenmann, 1907), G. balzanii (Perugia 1891), G. caaguazuensis Staeck, 2006, G. constellatus Malabarba, Malabarba & Reis, 2015, G. labiatus (Hensel, 1870), G. la- custris Reis & Malabarba, 1988, G. lipokarenos Malabarba, Malabarba & Reis, 2015, G. mekinos Malabarba, Malabarba & Reis, 2015, G. missioneiro Malabarba, Malabarba & Reis, 2015, G. pseudolabiatus Malabarba, Malabarba & Reis, Cichlidae constitutes one of the major vertebrate fami- lies, with more than 1,700 species (Fricke et al., 2018). The genus Gymnogeophagus Ribeiro is included in the tribe Geophagini (López-Fernández et al., 2005a, 2005b, 2010) along with approximately 14 other genera of the Neotropical subfamily Cichlinae. Species belonging to Gymnogeopha- gus are easily recognized by sharing two synapomorphies (Reis, Malabarba, 1988): the absence of supraneurals, and the presence of a forward-directed spine in the first pterygio phore of the dorsal fin (Fig. 1). Gymnogeophagus includes one extinct species, G. eocenicus Malabarba, Malabarba & Del Papa, 2010, known from fossil records of the Eocene Lumbrera Formation in northwestern Argentina, and 18 extant species from Paraná, Paraguay and Uruguay basins and small coastal drainages of Uruguay and southern Brazil, with one species, G. balzanii, also occurring in the rio Gua- poré, Amazon drainage (Malabarba et al., 2015; Loureiro et al., 2016; Casciotta et al., 2017a). A brief review of the taxonomic and phylogenetic works dealing with Gymnogeophagus was given by Malabarba et al. (2015) and is not repeated here. Figs. 1-5 Holotype. ZVC-P 12493, male, 101.9 mm SL, Uruguay, Salto, arroyo de las Tunas on Ruta 31, tributary to río Ar- apey Grande, 31°20’4.87”S 57°19’36.42”W, 8 Sep 2005, V. Bertaco, F. Cantera, J. Ferrer & L. R. Malabarba. Holotype. ZVC-P 12493, male, 101.9 mm SL, Uruguay, Salto, arroyo de las Tunas on Ruta 31, tributary to río Ar- apey Grande, 31°20’4.87”S 57°19’36.42”W, 8 Sep 2005, V. Bertaco, F. Cantera, J. Ferrer & L. R. Malabarba. Paratypes. Uruguay, Departamento de Salto. Río Arapey drainage: MHNM 3711, 4, 1 male 79.9 mm SL, 3 females or juveniles 50.0-69.5 mm SL, río Arapey Chico on Ruta 4, 31°2’7.44”S 56°53’50.21”W, 22 Nov 2005, F. Teixeira, A. D’Anatro, I. González, S. Oviedo & M. Loureiro. UFRGS 8042, 7, 2 males 93.1-106.1 mm SL and 5 unsexed juveniles 19.7-64.1 mm SL, tributary stream to río Arapey Grande on Ruta 4, 31°07’44.0”S 56°59’57.0”W, 8 Sep 2005, V. Bertaco, F. Cantera, J. Ferrer & L. R. Malabarba. UFRGS 8076, 14, 3 males 99.7-100.1 mm SL (1 c&s 74.2 mm SL), 2 females 74.3-77.2 mm SL and 9 unsexed juveniles 51.2-67.1 mm SL (1 c&s 40.8 mm SL), collected with the holotype. UFRGS 8101, 1, female 70.3 mm SL, stream on Ruta 4, tributary of stream Valentin Grande, 31°16’32.0”S 57°09’22.0”W, 8 Sep 2005, L. R. Malabarba, V. Bertaco, J. Ferrer & F. Cantera. ZVC-P 13084, 4, 2 males, 76.4-79.7 mm SL, 2 females or juveniles 46.4-55.6 mm SL, Cañada de la Tapera, tributary of río Arapey, 31°09’02.15”S 56°13’25.18”W, 13 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P Paratypes. Uruguay, Departamento de Salto. Río Arapey drainage: MHNM 3711, 4, 1 male 79.9 mm SL, 3 females or juveniles 50.0-69.5 mm SL, río Arapey Chico on Ruta 4, 31°2’7.44”S 56°53’50.21”W, 22 Nov 2005, F. Teixeira, A. D’Anatro, I. González, S. Oviedo & M. Loureiro. UFRGS 8042, 7, 2 males 93.1-106.1 mm SL and 5 unsexed juveniles 19.7-64.1 mm SL, tributary stream to río Arapey Grande on Ruta 4, 31°07’44.0”S 56°59’57.0”W, 8 Sep 2005, V. Bertaco, F. Cantera, J. Ferrer & L. R. Malabarba. UFRGS 8076, 14, 3 males 99.7-100.1 mm SL (1 c&s 74.2 mm SL), 2 females 74.3-77.2 mm SL and 9 unsexed juveniles 51.2-67.1 mm SL (1 c&s 40.8 mm SL), collected with the holotype. Figs. 1-5 UFRGS 8101, 1, female 70.3 mm SL, stream on Ruta 4, tributary of stream Valentin Grande, 31°16’32.0”S 57°09’22.0”W, 8 Sep 2005, L. R. Malabarba, V. Bertaco, J. Ferrer & F. Cantera. ZVC-P 13084, 4, 2 males, 76.4-79.7 mm SL, 2 females or juveniles 46.4-55.6 mm SL, Cañada de la Tapera, tributary of río Arapey, 31°09’02.15”S 56°13’25.18”W, 13 Dec 2014, M. Loureiro, A. Duarte, S. Serra, J. Bessonart & S. Paullier. ZVC-P e180118[2] e180118[2] A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Neotropical Ichthyology, 16(4): e180118, 2018 3 Neotropical Ichthyology, 16(4): e180118, 2018 A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Neotropical Ichthyology, 16(4): e180118, 2018 3 A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Diagnosis. The new species can be distinguished from the species of the Gymnogeophagus rhabdotus group and from G. balzanii by the shape of the caudal peduncle longer than deep (vs. deeper than long). It is distinguished from all con- geners, except G. labiatus and G. pseudolabiatus, by the possession of thick lips. It differs from G. labiatus and G. Diagnosis. The new species can be distinguished from the species of the Gymnogeophagus rhabdotus group and from G. balzanii by the shape of the caudal peduncle longer than deep (vs. deeper than long). It is distinguished from all con- geners, except G. labiatus and G. pseudolabiatus, by the possession of thick lips. It differs from G. labiatus and G. lacustris by the lack of an oblique bar from the eye to the dorsal-fin origin (vs. oblique bar present), and by the co- lor pattern of the caudal, dorsal and anal fins with dots (vs. caudal fin and posterior portion of anal fin with longitudi- nal hyaline stripes). It differs from G. pseudolabiatus and G. mekinos by the hump entirely black in males (Fig. 2; vs. Fig. 2. Head of Gymnogeophagus peliochelynion (first column), G. pseudolabiatus (second column) and G. mekinos (third column) showing the entirely black hump in males in G. peliochelynion (vs. yellow with black margin), and upper lip not folded dorsally over anterior margin of snout (vs. upper lip folded dorsally in G. pseudolabiatus and undeveloped in G. me- kinos). G. peliochelynion from top to bottom, paratype, ZVC-P 13210, paratype, 76.3 mm SL; ZVC-P 7016, 89.9 mm SL; ZVC-P 13057, 90.2 mm SL. G. Figs. 1-5 Prepelvic contour straight to slightly con- vex; abdominal contour straight and base of anal fin straight to slightly convex. Dorsal-fin spines 12(1), 13(2), 14(11); dorsal-fin soft rays 9(1), 10(11), 11(2). First dorsal-fin spine inserted right above vertical line across posterior bony margin of opercle. Soft dorsal fin slightly pointed in young and adult females, reaching or almost reaching caudal-fin base. Fourth or fifth dorsal-fin soft ray longest in mature males, reaching proximal two third to half of caudal-fin length. Three anal- fin spines; anal-fin soft rays 8(11), 9(3); anal-fin profile rounded in young and females, reaching or almost reaching caudal-fin base; slightly pointed in reproductive males, sur- passing caudal-fin base. Pectoral fin with pointed tip, reach- ing or almost reaching anal-fin origin in juveniles and fe- males and surpassing anal-fin origin in reproductive males. Pelvic fin slightly pointed; second soft ray longest, reaching anal-fin base in mature males. Caudal-fin margin concave. Tab. 1. Morphometric data of Gymnogeophagus peliochely- nion, new species. Standard length is expressed in mm. Range includes measurements of holotype and 22 paratypes, except individuals smaller than 45 mm SL. Tab. 1. Morphometric data of Gymnogeophagus peliochely- nion, new species. Standard length is expressed in mm. Range includes measurements of holotype and 22 paratypes, except individuals smaller than 45 mm SL. Holotype Min Max Mean SD Standard length (mm) 101.9 46.42 106.08 73.87 - Percent of standard length Head length 38.3 35.0 39.2 36.8 1.07 Body depth 38.5 33.8 39.9 37.7 1.44 Dorsal-fin base length 48.2 45.4 54.1 50.3 2.05 Pectoral-fin length 33.6 30.1 39.3 34.4 3.19 Caudal peduncle depth 13.2 11.9 14.2 13.3 0.52 Caudal peduncle length 15.1 14.0 18.1 15.3 1.10 Percent of head length Eye diameter 21.3 20.2 32.3 25.1 3.27 Interorbital width 28.5 24.6 32.4 28.0 1.97 Upper jaw length 26.7 19.7 29.7 23.5 3.39 Pre-orbital length 33.4 26.9 36.6 30.7 2.83 Snout length 57.7 30.5 59.7 46.4 7.64 ii g Jaw teeth small, conical, with recurved tips. Upper jaw with outer regular row of 16-26 teeth in each premaxilla (number increasing with specimen size) and two irregu- lar internal rows of slightly smaller teeth. Lower jaw with 3-5 irregular rows of small conical teeth; outer hemiseries with 20-28 teeth. Lower limb of first gill arch with 6-10 gill rakers; upper limb lobed with 3-4 gill rakers in its margin. Lower pharyngeal tooth plate wide; teeth covering whole occlusion surface. Figs. 1-5 pseudolabiatus from top to bottom, paratype, UFRGS 7754, 102.0 mm SL; MHNM 4010, 88.8 mm SL; MHNM 4010, 95.3 mm SL. G. mekinos from top to bottom, MHNM 3511, 105.1 mm SL; MHNM 3511, 97.2 mm SL; MHNM 4009, 121.3 mm SL. Fig. 2. Head of Gymnogeophagus peliochelynion (first column), G. pseudolabiatus (second column) and G. mekinos (third column) showing the entirely black hump in males in G. peliochelynion (vs. yellow with black margin), and upper lip not folded dorsally over anterior margin of snout (vs. upper lip folded dorsally in G. pseudolabiatus and undeveloped in G. me- kinos). G. peliochelynion from top to bottom, paratype, ZVC-P 13210, paratype, 76.3 mm SL; ZVC-P 7016, 89.9 mm SL; ZVC-P 13057, 90.2 mm SL. G. pseudolabiatus from top to bottom, paratype, UFRGS 7754, 102.0 mm SL; MHNM 4010, 88.8 mm SL; MHNM 4010, 95.3 mm SL. G. mekinos from top to bottom, MHNM 3511, 105.1 mm SL; MHNM 3511, 97.2 mm SL; MHNM 4009, 121.3 mm SL. e180118[3] Neotropical Ichthyology, 16(4): e180118, 2018 4 New Gymnogeophagus from lower río Uruguay New Gymnogeophagus from lower río Uruguay dal fin with small elongated cycloid scales in single series between contiguous rays. Dorsal fin without scales. Cheek naked. Small cycloid scales on opercle. Subopercular scales ctenoid in 1-2 irregular rows. Ctenoid scales on base of hump (in adult reproductive males only) to approximately vertical line passing on anterior margin of eye. Scales in longitudinal series 26(5), 27(6), 28(3). Anterior lateral line 15(1), 17(8), 18(5). Posterior lateral line 8(4), 9 (6), 10(1), 11(2), 12(1). Scales between anterior lateral line and dorsal fin 6(12), 7(2). Scales between anterior lateral line and anal fin 8(11), 9(3).ii yellow with black margin), and upper lip not folded dorsally over anterior margin of snout (vs. upper lip folded dorsally, usually with a well-developed medial lobe dorsally projec- ted in G. pseudolabiatus). Description. Standard length of specimens examined 19.2 to 106.1 mm. Morphometric data summarized in Tab. 1. Body elongated, laterally compressed. Dorsal profile of head slightly convex between mouth and interorbital area in young and females, slightly straight in males; slightly convex from interorbital region to dorsal-fin origin. Reproductive males with adipose hump from interorbital region to dorsal-fin origin; dorsal-fin base gently convex. Caudal peduncle rec- tangular, longer than deep, with slightly concave dorsal and ventral profiles. Figs. 1-5 Teeth on medial rows larger than remain- ing ones. Posterolateral teeth elongated; posteromedial teeth larger, cylindrical with medial, blunt cusps of molariform aspect (Fig. 3). Coloration after fixation in formalin. (Fig. 4) Mature ma- les: ground color of body dark brown above longitudinal series of scales of posterior lateral line and light yellowish brown below. Series of double vertical bars clearly discerni- ble along midventral lateral surface of flanks, distributed be- tween pectoral-fin base and end of caudal peduncle, anterior to caudal-fin base. Number of vertical double dark bars 5-6. Midlateral spot without defined borders. Hump entirely black in males making dark band in front of dorsal-fin origin hardly detectable. Head dark brown with some black spots on cheek and near posterior margin of opercle; dark band covering cheek hardly distinct. Isthmus and branchiostegal membranes dark brown. Pectoral fin hyaline. Pelvic fin dark gray. Spi- nous dorsal fin dark brown; soft dorsal fin light brown with circular dots. Distal one third of anal-fin rays and spines dark brown, without additional marks. Two proximal thirds of anal-fin rays covered with small circular dots. Caudal fin co- vered with white dots, except near upper and lower borders. Head depth larger than head length. Snout triangular in lateral aspect; slightly rounded anteriorly, pointed in dorsal aspect. Eyes small, close to dorsal profile of head in juveniles and progressively farther from it in larger specimens (about eye diameter in specimens up to 80 mm); eyes near middle of head length. Interorbital area slightly convex in young and female, and deeply convex in large males; interorbital width larger than eye diameter, except in individuals smaller than 25 mm. Mouth terminal. Posterior tip of maxilla not reaching vertical line across anterior margin of eyes. Upper jaw slightly longer than lower jaw; lips developed, lower lip thicker than upper lip; margin of lower lip convex, deeper in middle length of each dentary, deeply notched medially at symphysis. Snout longer than postorbital length, except in individuals up to 48 mm SL. Body scales moderately large and ctenoid, smaller cte- noid scales in preventral area. Proximal third to half of cau- e180118[4] e180118[4] Neotropical Ichthyology, 16(4): e180118, 2018 Neotropical Ichthyology, 16(4): e180118, 2018 5 Neotropical Ichthyology, 16(4): e180118, 2018 A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Color in alcohol of preserved females and young not distinct from that described for males. Main differences are: isthmus and branchiostegal membrane yellow; clearly distinct dark band covering cheek, below eye; distinct dark band on dorsal-fin origin; and larger size of white spots on dorsal, anal, and caudal fins. Fig. 3. Pharyngeal tooth plate of Gymnogeophagus pelio- chelynion, 74.2 mm SL, UFRGS 8076, paratype. Dorsal view; anterior to bottom. Coloration in life. (Fig. 5) Ground color of dorsal profile golden or dark olivaceous with longitudinal series of light blue spots. Well defined black, circular midlateral spot, cove- ring scales 9-11 of the anterior lateral line, scales 10-12 of the scale row just below anterior lateral line and scales 8-10 of the next scale row below. Large dark bar below eye, usually not reaching midline of mouth in larger individuals. Nume- rous small bright blue spots usually present on cheeks. Red marks on cheeks usually present in mature males, concentra- ted on upper portion of opercle and preopercle, region behind eye and above pectoral fin. Usually grayish blue lips in repro- ductive males and gray in females and young. Adipose hump, when present, black. Ventral portion of body light olivaceous to yellow pale in mature males with longitudinal series of Fig. 3. Pharyngeal tooth plate of Gymnogeophagus pelio- chelynion, 74.2 mm SL, UFRGS 8076, paratype. Dorsal view; anterior to bottom. Fig. 4. Gymnogeophagus peliochelynion: top, holotype, male, ZVC-P 12493, 101.9 SL; bottom, paratype, female, UFRG 8076, 77.2 SL. Both from arroyo de las Tunas on road 31, tributary of río Arapey Grande, Salto, Uruguay. Photographs take just after collection and fixation in formalin. Fig. 4. Gymnogeophagus peliochelynion: top, holotype, male, ZVC-P 12493, 101.9 SL; bottom, paratype, female, UFRGS 8076, 77.2 SL. Both from arroyo de las Tunas on road 31, tributary of río Arapey Grande, Salto, Uruguay. Photographs taken just after collection and fixation in formalin. e180118[5] Neotropical Ichthyology, 16(4): e180118, 2018 New Gymnogeophagus from lower río Uruguay New Gymnogeophagus from lower río Uruguay Fig. 5. Gymnogeophagus peliochelynion: above, paratype, male, ZVC-P 13210, paratype, 76.3 mm SL, río Arapey, Colonia Lavalleja, Paso Elías, Salto, Uruguay; below, female, ZVC-P 13057, 65.3 mm SL, Arroyo Sopas, Paso del Cementerio, Salto, Uruguay. Photographs of live specimens. Fig. 5. Neotropical Ichthyology, 16(4): e180118, 2018 Gymnogeophagus peliochelynion: above, paratype, male, ZVC-P 13210, paratype, 76.3 mm SL, río Arapey, Colonia Lavalleja, Paso Elías, Salto, Uruguay; below, female, ZVC-P 13057, 65.3 mm SL, Arroyo Sopas, Paso del Cementerio, Salto, Uruguay. Photographs of live specimens. Etymology. The name peliochelynion is from the Greek pe- lios, meaning black and blue, and chelyne, meaning lip, in reference to the color of the lips of the new species. A name in apposition. Etymology. The name peliochelynion is from the Greek pe- lios, meaning black and blue, and chelyne, meaning lip, in reference to the color of the lips of the new species. A name in apposition. light blue spots. Spinous dorsal fin and base of soft dorsal fin yellowish brown; most soft dorsal fin red with relatively lar- ge and numerous hyaline dots. Distal tip of dorsal fin hyaline. Pectoral fin hyaline and pelvic fin dark orange to dark gray with light blue spots at base. Anal fin yellowish orange pro- ximally with numerous clear spots and hyaline on half distal portion with dark gray margin. Caudal fin yellowish brown with numerous light spots, extending along middle of caudal fin; dorsal and ventral portions of fin hyaline. Conservation status. Gymnogeophagus peliochelynion is relatively frequent and abundant in the río Arapey Grande drainage. No specific threats were detected, and the spe- cies can be categorized as Least Concern (LC) according to IUCN criteria (IUCN, 2016). Geographic distribution. This species is known from the río Arapey Grande drainage, a tributary of the lower río Uru- guay basin, Uruguay (Fig. 6). Discussion Gymnogeophagus peliochelynion shares the two sy- napomorphies that diagnoses the genus (Reis, Malabar- ba, 1988): the lack of supraneurals and the presence of a forward-directed spine on anterodorsal margin of the first Ecological notes. The new species was collected in rivers with clear water, usually with rocky or muddy bottom and little vegetation. e180118[6] e180118[6] Neotropical Ichthyology, 16(4): e180118, 2018 A. Turcati, W. S. Serra-Alanis & L. R. Malabarba A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Fig. 6. Geographic distribution of Gymnogeophagus peliochelynion (yellow; star refers to type-locality), G. cf. peliochely- nion (orange) and other species of the Gymnogeophagus gymnogenys clade: G. lipokarenos (blue), G. constellatus (green), G. missioneiro (red), G. tiraparae (white), G. mekinos (black), G. pseudolabiatus (pink) in the río Uruguay basin. Records of Gymnogeophagus lipokarenos and Gymnogeophagus constellatus from Argentina (triangles) are based on Casciotta et al. (2017b) and Říčan et al. (2017), respectively. Fig. 6. Geographic distribution of Gymnogeophagus peliochelynion (yellow; star refers to type-locality), G. cf. peliochely- nion (orange) and other species of the Gymnogeophagus gymnogenys clade: G. lipokarenos (blue), G. constellatus (green), G. missioneiro (red), G. tiraparae (white), G. mekinos (black), G. pseudolabiatus (pink) in the río Uruguay basin. Records of Gymnogeophagus lipokarenos and Gymnogeophagus constellatus from Argentina (triangles) are based on Casciotta et al. (2017b) and Říčan et al. (2017), respectively. arenos is endemic to the upper portion of the río Uruguay, upstream to San Javier (Argentina)/Porto Xavier (Brazil). Gymnogeophagus constellatus and G. missioneiro occur in tributaries immediately downstream of San Javier and Porto Xavier, the first occurring mainly in the rio Ijuí (Bra- zil), arroyo Itacaruaré and arroyo Chimiray-Miní (Misiones) drainages, and the second occurring in rio Piratini (Brazil) drainage, all draining a basaltic rock bed of the Serra Geral Formation (Batezelli et al., 2005). The next main tributary of the Río Uruguay, in a downstream direction, is the rio Ibicuí, with a fine grain sedimentary substrate, correspond- ing to the unique tributary of the middle Río Uruguay where G. tiraparae and G. mekinos are found. These two species are present too in lower tributaries of Uruguay and La Plata river basins in Uruguay and Brazil. In the next downstream drainage, the rio Quaraí, with basaltic rock bed, we find G. pseudolabiatus. The next downstream tributary, the río Ar- apey, is occupied by the new species described herein, G. peliochelynion. This successive substitution of allopatric dorsal-fin pterygiophore (Fig. 1). Discussion The new species also belong to a clade that includes G. gymnogenys. Species in this clade are easily recognized by sharing two synapomor- phies (more easily observed in females and juveniles; Fig. 5): the absence of an oblique bar between the dorsal border of the eye and the nape, and the possession of a black bar originating in the dorsal contour near the dorsal-fin origin and directed downward and backward on the dorsum (Reis, Malabarba, 1988; Malabarba et al., 2015). The conspicuous secondary sexual dimorphism (including development of a nuchal hump in reproductive males), mouthbrooding repro- ductive strategy, and elongated caudal peduncle, longer than deep (Wimberger et al., 1998), which are not observed in the G. rhabdotus group, further support the inclusion of G. peliochelynion in the G. gymnogenys clade. Species of the Gymnogeophagus gymnogenys clade have an allopatric distribution (Fig. 6) along several tribu- taries of the río Uruguay (Malabarba et al., 2015; Casciotta et al., 2017b; Říčan et al., 2017). Gymnogeophagus lipok- e180118[7] Neotropical Ichthyology, 16(4): e180118, 2018 8 New Gymnogeophagus from lower río Uruguay ra, I. González & F. Quintans; ZVC-P 10011, 11, 24.8-87.4 mm SL, Cañada Honda, 30°27’46.98”S 56°51’24.82”W, Aug 2006, M. Lou- reiro, F. Teixeira, I. González & F. Quintans; ZVC-P 10057, 27, 28.1- 80.9 mm SL, Cañada Honda, 30°27’33.47”S 56°49’51.29”W, Aug 2006, M. Loureiro, F. Teixeira, I. González & F. Quintans; ZVC-P 12616, 7, 46.6-93.2 mm SL, Cuareim River, Laguna Salamanca, 30°46’36.21”S 56°02’16.71”W, Dec 2012, A. Duarte, W.S. Serra, M. Loureiro & L. Ziegler; ZVC-P 12684, 2, 98.2-101.6 mm SL, Cuaró Grande stream, Ruta 4, 30°46’57.96”S 56°47’05.08”W, Aug 2013, M. Loureiro, J. Bessonart, W.S. Serra & L. Montes de Oca. species of the Gymnogeophagus gymnogenys clade along río Uruguay tributaries suggests the main channel act as a physical or ecological barrier to dispersal of these species, all of which are adapted to small affluents. Similar patterns have been observed in the tributaries of the rio Amazonas for other groups of fish (Junk et al., 2001; Hubert, Renno 2006) and for Atlantic coastal rivers connected though freshwater lakes (Hirschmann et al., 2015). Populations from río Queguay and río Dayman located downstream from the río Arapey are tentatively identi- fied herein as G. cf. peliochelynion. Although they do not show hypertrophied lips, they show similar color pattern to that described for G. peliochelynion. Acknowledgments We are grateful to the following colleagues and their institutions for the loan of specimens: Carlos A. S Lucena (MCP) and Marcelo Loureiro (ZVC-P). We are grateful to Néstor Ríos, Cristhian Clavijo, Alejandro Duarte, Marcelo Loureiro, Sofía Paulier, Mauricio Pérez, Juliano Ferrer, Vinícius Bertaco, and Felipe Cantera for their assistance with fieldwork and to Mayara Neves for the photo of Fig. 3. LRM is partially financed by CNPq (307890/2016-3; 401204/2016-2). Comparative material examined (In addition to those listed by Malabarba et al., 2015). Gymnogeophagus cf. peliochelynion. Uruguay, Departamento de Salto. Río Dayman drainage: ZVC-P 6686, 1, 57.7 mm SL, río Dayman, Ruta 4, 31°47’15.45”S 57°02’12.24”W, 21 Nov 2005, M. Loureiro, F. Teixeira, S. Ovie- do, A. D’Anatro & I. González. ZVC-P 13242, 1, 54.2 mm SL, Río Dayman, Paso Morales, 31°28’40.92”S 57°49’40.14”W, 17 Dec 2014, M. Loureiro, S. Serra & A. Duarte. Departamento de Paysandú. Río Queguay drainage: ZVC-P 13299, 1, 53.7 mm SL, Río Queguay, Paso del Sauce, 32°16’55.46”S 56°45’17.24”W, 19 Dec 2014, A. Duarte, S. Serra & M. Loureiro. ZVC-P 13306, 1, 70.5 mm SL, Arroyo Corrales, Paso Castillo, 31°56’29.54”S 56°37’12.40”W, 19 Dec 2014, A. Duarte, S. Serra & M. Loureiro. ZVC-P 13348, 1, 48.5 mm SL, Río Queguay, Cerro del Inglés, 32°10’31.73”S 57°44’00.01”W, 19 Apr 2015, S. Paullier, J. Cabre- ra, M. Trillo, M. Loureiro & J. Bessonart. ZVC-P 13367, 1, 108.3 mm SL, Arroyo Ñacurutú Grande, 32°18’39.78”S 57°24’20.88”W, 21 Apr 2015, S. Paullier, J. Cabrera, M. Trillo, M. Loureiro & J. Bessonart. ZVC-P 13387, 1, 65.8 mm SL, Río Queguay Chico, Paso del Parque, 32°02’27.76”S 56°59’26.24”W, Apr 2015, S. Paullier, J. Cabrera, M. Trillo, M. Loureiro & J. Bessonart. ZVC-P 13439, 1, 67.7 mm SL, Arroyo Bacacuá Grande, 32°19’12.30”S 57°36’16.66”W, 21 Apr 2015, S. Paullier, J. Cabrera, M. Trillo, M. Loureiro & J. Bessonart. Gymnogeophagus gymnogenys. Bra- zil, Rio Grande do Sul State (all from laguna dos Patos draina ge): UFRGS 4242, 10, 47.3-65.4 mm SL, arroio Velhaco; UFRGS 13697, 1, 106.3 mm SL, arroio Forquetinha; UFRGS 14213, 8, 39.9-94.8 mm SL, arroio Forquetinha; UFRGS 16135, 1, 88.1 mm SL, lago Guaíba; UFRGS 16136, 1, 92.4 mm SL, lago Guaíba. Gymnogeophagus mekinos. Uruguay, Departamento de Flori- da. MHNM 3511, 2, 97.2-105.1 mm SL, Paso de la Arena, Río Santa Lucía Chico, 34°01’51.24”S 56°09’45.75”W, 11 Jan 2014, W.S. Serra & M. Pérez. Departamento de Rocha. Acknowledgments MHNM 4009, 4, 49.9-121.3 mm SL, Arroyo Rocha, Parque La Estiva, Ciudad de Rocha, 34°29’38.43”S 54°20’43.58”W, 22 Jan 2018, W.S. Ser- ra. Gymnogeophagus pseudolabiatus. Uruguay, Departamento de Artigas. MHNM 4010, 7, 55.3-95.3 mm SL, Arroyo Pintado y Ruta 30, 30°26’16.70”S 56°26’59.23”W, 2 Dec 2017, W.S. Serra & N. Ríos; ZVC-P 9893, 5, 34.9-88.2 mm SL, Cañada Mataojo, 30°47’46.93”S 56°56’52.66”W, Aug 2006, M. Loureiro, F. Teixei- Discussion Further investigation is needed in order to determine whether these populations constitute a separate species. References Batezelli A, Saad AR, Fulfaro VJ, Corsi AC, Landim PMB, Perinotto JAJ. Análise de bacia aplicada às unidades mesozoicas do triângulo mineiro (sudeste do Brasil): uma estratégia na prospecção de recursos hídricos subterrâneos. Águas Subterrâneas. 2005; 19(1):61-73. Casciotta J, Almirón A, Piálek L, Říčan O. Gymnogeophagus taroba (Teleostei: Cichlidae), a new species from the río Iguazú basin, Misiones, Argentina. Historia Natural, Tercera Serie. 2017a; 7(2):5-22. Casciotta J, Almirón A, Říčanová Š, Dragová K, Piálek L, Alonso F, Říčan O. First record of Gymnogeophagus lipokarenos Malabarba, Malabarba & Reis, 2015 (Teleostei: Cichliformes) from Argentina. Ichthyol Contrib PecesCriollos 2017b; 54:1-5. Fricke R, Eschmeyer WN, Fong JD. Species by family/subfamily in the Catalog of Fishes. [Electronic version]. San Francisco (CA): California Academy of Sciences; 2018. [cited 2018 Sep 6]. Available from: http://researcharchive.calacademy.org/ research/ichthyology/catalog/SpeciesByFamily.asp González-Bergonzoni I, Loureiro M, Oviedo S. A new species of Gymnogeophagus from the río Negro and río Tacuarí basins, Uruguay (Teleostei: Perciformes). Neotrop Ichthyol. 2009; 7(1):19-24. Hirschmann A, Malabarba LR, Thomaz AT, Fagundes NJR. Riverine habitat specificity constrains dispersion in a Neotropical fish (Characidae) along Southern Brazilian drainages. Zool Scr. 2015; 44(4):374-82. Hubert N, Renno F. Historical biogeography of South American freshwater fishes. J Biogeogr. 2006; 33(8):1414-36. International Union for Conservation of Nature (IUCN). Standards and petitions subcommittee. Guidelines for using the IUCN e180118[8] e180118[8] Neotropical Ichthyology, 16(4): e180118, 2018 A. Turcati, W. S. Serra-Alanis & L. R. Malabarba Miranda Ribeiro, 1918 (Teleostei: Cichliformes) from the rio Uruguay drainage. Neotrop Ichthyol. 2015; 13(4):637-62. red list categories and criteria. Version 12 [Internet]. 2016. Available from: http://www.iucnredlist.org/documents/ RedListGuidelines.pdf/ Miranda Ribeiro, 1918 (Teleostei: Cichliformes) from the rio Uruguay drainage. Neotrop Ichthyol. 2015; 13(4):637-62. red list categories and criteria. Version 12 [Internet]. 2016. Available from: http://www.iucnredlist.org/documents/ RedListGuidelines.pdf/ Reis RE, Malabarba LR. Revision of the Neotropical genus Gymnogeophagus Ribeiro, 1918, with descriptions of two new species (Pisces, Perciformes). Rev Bras Zool. 1988; 4(4):259- 305. Junk WJ, Soares MGM. Freshwater fish habitats in Amazonia: state of knowledge, management, and protection. Aquat Ecosyst Health Manag. 2001; 4(4):437-51. Říčan O, Piálek L, Říčanová Š, Dragová K, Almirón A, Casciotta J. First record of Gymnogeophagus constellatus Malabarba, Malabarba & Reis, 2015 (Teleostei: Cichliformes) from Argentina. Ichthyol Contrib PecesCriollos. 2017; 56:1-3. López-Fernández H, Honeycutt RL, Stiassny ML, Winemiller KO. Morphology, molecules, and character congruence in the phylogeny of South American geophagine cichlids (Perciformes, Labroidei). Zool Scr. 2005a; 34(6):627-51. López-Fernández H, Honeycutt RL, Winemiller KO. Submitted September 8, 2018 Accepted October 14, 2018 by William Crampton References Molecular phylogeny and evidence for an adaptive radiation of geophagine cichlids from South America (Perciformes: Labroidei). Mol Phylogenet Evol. 2005b; 34(1):227-44. Říčan O, Říčanová Š, Dragová K, Piálek L, Almirón A, Casciotta J. Species diversity in Gymnogeophagus (Teleostei: Cichlidae) and comparative biogeography of cichlids in the Middle Paraná basin, an emerging hotspot of fish endemism. Hydrobiologia. 2018; 1-24. López-Fernández H, Winemiller KO, Honeycutt RL. Multilocus phylogeny and rapid radiations in Neotropical cichlid fishes (Perciformes: Cichlidae: Cichlinae). Mol Phylogenet Evol. 2010; 55(3):1070-86. Wimberger PH, Reis RE, Thornton K. Mitochondrial phylogenetics, biogeography, and evolution of parental care and mating systems in Gymnogeophagus (Perciformes: Cichlidae). In: Malabarba LR, Reis RE, Vari RP, Lucena ZMS, Lucena CAS, editors. Phylogeny and classification of Neotropical fishes. Porto Alegre: Edipucrs; 1998. p.509-18. Wimberger PH, Reis RE, Thornton K. Mitochondrial phylogenetics, biogeography, and evolution of parental care and mating systems in Gymnogeophagus (Perciformes: Cichlidae). In: Malabarba LR, Reis RE, Vari RP, Lucena ZMS, Lucena CAS, editors. Phylogeny and classification of Neotropical fishes. Porto Alegre: Edipucrs; 1998. p.509-18. Loureiro M, Zarucki M, Malabarba LR, González-Bergonzoni I. A new species of Gymnogeophagus Miranda-Ribeiro from Uruguay (Teleostei: Cichliformes). 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https://openalex.org/W4394959285	OpenAlex	Open Science	CC-By-SA	null	Examining the Connection between Service Failure and Recovery with Airline Passenger Satisfaction with Air Asia	. Aarti	English	Spoken	4,737	7,807	Abstract A firm's service failure and the way it addresses it can have a significant impact on customer satisfaction and their overall relationship with the company. This is true even when the company has put in substantial efforts to build strong, long-term relationships with its customers. Airline companies, in particular, face various challenges that affect their survival over time. One of these challenges is dealing with inevitable service failures. The goal of this study is to understand how service failure and recovery relate to passenger satisfaction specifically within AirAsia. Data was collected from 361 domestic travelers departing from Sultan Ismail Petra Airport in Kelantan. Convenience sampling was used for this study. The results showed that there is a significant negative correlation between service failure and passenger satisfaction, while service recovery has a positive impact on passenger satisfaction. The implications suggest that the management team and staff should implement effective recovery strategies to address failures, including well-planned corrective actions, sincere apologies, taking responsibility for any mishaps, and other measures. Frontline employees need to be attuned to customers' emotions during service failures; effective recovery efforts can help prevent negative customer reactions. Keywords: Service Failure, Service Recovery, Customer Satisfaction, Relationship Quality, Airline Companies Examining the Connection between Service Failure and Recovery with Airline Passenger Satisfaction with Air Asia Aarti Student of BBA in Aviation Management, School of Business, Galgotias University 1. INTRODUCTION Nowadays, marketers are focused on improving customer satisfaction. Numerous studies have emphasized the significance of satisfaction in the service industry (Ali et al., 2020). Customer satisfaction is affected by the quality of service. Service failures are often obstacles to achieving customer satisfaction, and they are difficult to avoid within the service delivery process (Sukri, 2014). Customers' increasing expectations and the competitive business landscape make it challenging for many companies, particularly airlines, to retain their customers (Wang et al., 2011). ” Another challenge in providing excellent services is the inability to recognize the genuine needs and desires of customers (Izogo & Ogba, 2015). Customers often express disappointment, end their patronage, or spread negative sentiments after experiencing service failures. Therefore, it is essential to focus on delivering outstanding service recovery in order to restore a positive perception among customers (Daşkın & Yılmaz, 2015). Numerous researchers have conducted studies on service failure in different industries. Scholars also focus extensively on the effects and impact of service failure and recovery in the airline industry. Volume 6, Issue 2, March-April 2024 IJFMR240217568 1 1 rnational Journal for Multidisciplinary Research (IJFMR) Furthermore, service failures can have a detrimental impact on customer satisfaction, which in turn affects the company's reputation (Ali et al., 2020). Service failures are a common occurrence in many companies and can result in dissatisfied customers. Several airlines, including well-known international ones like Champion Air, Aloha Airgroup, British regional airline Flybmi, and others have either stopped operating or gone out of business (Insider, 2019). During the 2018-2019 fiscal year, numerous airline companies went out of business, including Air2there (a New Zealand carrier), Air Costa Rica, NextJet, and several others (Izogo & Ogba, 2015). This arises from numerous challenges faced, such as inadequate management of operations where the leadership is unable to prevent service failures and address problems effectively. Additionally, high labor costs and jet fuel expenses have been a major source of frustration for many airlines (Izogo & Ogba, 2015). Malaysia's national carrier has faced challenges over the past few years. According to aviation experts, MAS is also dealing with significant internal management issues following the tragedies involving two high-profile planes, MH370 and MH17 (Namukasa, 2013). (Curry & Gao, 2012) Customers who are satisfied with the recovery effort have a more positive outlook than those who are not. It is crucial for airline operations to effectively recover from failures, and managers must understand customer feedback on these issues (Curry & Gao, 2012). Despite the extensive research on service quality, there is still limited exploration of service failure and recovery within the airline industry in Malaysia. Further investigation is needed (Sukri, 2014) (Ringle et al., 2011). The distinct characteristics of the airline service industry, compared to other service sectors, allow for additional research on understanding how service failures and recovery efforts impact customer satisfaction (Ringle et al., 2011). Previous research has identified several factors that contribute to service failures in the airline industry, such as challenges with the booking process, issues related to staff attitudes, trip cancellations, double- booking flights, and flight changes or delays (Ringle et al., 2011). There has been a growing number of reports of failure in the airline industry, suggesting that effective service recovery will receive positive responses from customers. Conducting a study on this issue is essential to identify overlooked problems and take further action to address this issue(Chou, 2015). “ 2.2 Service Recovery Transforming dissatisfied clients into happy ones involves effective service recovery (Ha & Jang, 2009). As stated by (Sukri, 2014), even when it's not possible to completely eliminate service failures, utilizing effective strategies for addressing and resolving these issues can help maintain, and possibly even enhance, customer satisfaction and loyalty over time. Studies have suggested that organizations can employ various methods to rectify failures, such as seeking feedback from customers through communication and offering explanations for the mistakes made (Chahal & Devi, 2015) and expressing regret for their shortcomings (Hu et al., 2013). Many researchers who study service quality have devoted significant attention to investigating the impact of recovery efforts on customer satisfaction and their likelihood to make repeat purchases. One important outcome is that effective service recovery can transform disappointed and frustrated customers into loyal ones (Cowart et al., 2014). 2.1. Service Failure Service failure happens when the service quality does not meet customer expectations (Chahal & Devi, 2015). Service failure often leads to customer attrition for many businesses. Neglecting this issue can result in decreased customer satisfaction and negative feedback from dissatisfied customers (Bankova, 2018). Previous research has identified various causes of service breakdowns in the airline sector, such as impolite staff, flight disruptions, reservation challenges, overbookings, diversions, and others (Ringle et al., 2011). Few studies have focused on reducing instances of service failure, as recommended by (Chahal & Devi, 2015), in organised service settings, the service failure is less likely to happen. (Cowart et al., 2014) further proposed that a positive communication approach and empowering employees will reduce the likelihood of failures. Further, (Steven et al., 2012) The causes of service failure have been categorized into two factors: customer and organization. Service failure occurs when errors are made by the service provider, an affiliated firm, or the customer themselves (Sukri, 2014). Consequences may occur following a breakdown, impacting customers' feelings and leading to emotions such as dissatisfaction, annoyance, and apathy. Many customers opt to abandon the service or switch their allegiance to a rival company without Volume 6, Issue 2, March-April 2024 IJFMR240217568 2 2 International Journal for Multidisciplinary Research (IJFMR) E-ISSN: 2582-2160 ● Website: www.ijfmr.com ● Email: editor@ijfmr.com voicing their grievances about previous failures. Failing to report the encountered issues makes it unlikely for customers to consider repurchasing from the company. voicing their grievances about previous failures. Failing to report the encountered issues makes it unlikely for customers to consider repurchasing from the company. 3. METHODOLOGY This research endeavors to delve into the intricate interplay between service failure and service recovery mechanisms and their collective impact on airline customer satisfaction. Focusing specifically on AirAsia's domestic passengers departing from Sultan Ismail Petra Airport in Kota Baharu Kelantan, where the monthly passenger population averages between 5000 to 6000 individuals, this study represents a targeted effort to explore the dynamics of service quality within a specific airline and airport context. To facilitate the investigation, convenience sampling was employed, given its pragmatic suitability for capturing a representative sample of passengers within the airport premises. Utilizing the renowned "Table: Determination of Sample Size" developed by (Krejcie & Morgan, 1970), the appropriate sample size of 361 respondents was determined, exceeding the required minimum for robust statistical analysis. Consequently, 372 questionnaires were distributed among eligible participants, ensuring adequate coverage of the intended population. This research endeavors to delve into the intricate interplay between service failure and service recovery mechanisms and their collective impact on airline customer satisfaction. Focusing specifically on AirAsia's domestic passengers departing from Sultan Ismail Petra Airport in Kota Baharu Kelantan, where the monthly passenger population averages between 5000 to 6000 individuals, this study represents a targeted effort to explore the dynamics of service quality within a specific airline and airport context. To facilitate the investigation, convenience sampling was employed, given its pragmatic suitability for capturing a representative sample of passengers within the airport premises. Utilizing the renowned "Table: Determination of Sample Size" developed by (Krejcie & Morgan, 1970), the appropriate sample size of 361 respondents was determined, exceeding the required minimum for robust statistical analysis. Consequently, 372 questionnaires were distributed among eligible participants, ensuring adequate coverage of the intended population. The survey questionnaire emerged as the primary instrument for data collection, meticulously designed to encapsulate key research inquiries pertinent to service failures, service recovery efforts, and passenger satisfaction as delineated in the study's overarching objectives. Administered directly within the departure hall, the questionnaire was strategically deployed to maximize respondent accessibility and engagement, thereby enhancing the reliability and validity of the data gathered. By leveraging the insights garnered from this comprehensive survey instrument, this study endeavors to unravel the nuanced dynamics shaping passenger perceptions and experiences within the context of service delivery in the airline industry. yp H0: There is a negative relationship between service recovery with passenger satisfaction H1: There is a positive relationship between service recovery with passenger satisfaction. H0: There is a negative relationship between service recovery with passenger satisfaction H1: There is a positive relationship between service recovery with passenger satisfaction. 3. METHODOLOGY Through rigorous analysis and interpretation of the collected data, the research aims to generate actionable insights and recommendations aimed at enhancing service quality, fostering customer satisfaction, and bolstering the competitive positioning of AirAsia within the dynamic aviation marketplace. “ 2.3 Customer Satisfaction Customer satisfaction is the experience a customer has with a service compared to their expectations. It is created when the service provider meets or surpasses those expectations. Many studies have examined whether satisfaction is influenced by service quality, and scholars have found a strong link between the two. All services should be geared towards enhancing consumer satisfaction (Nair et al., 2021). When customers' expectations are met, they often express satisfaction (Hussain et al., 2015). Satisfaction results in either a positive or negative sense of fulfillment. In today's highly competitive market, customer satisfaction has become an invaluable asset for the airline industry (Namukasa, 2013). In today's competitive airline industry, many airlines are under pressure and must act quickly to survive (Steven et al., 2012). A diverse range of services, such as excellent customer service, has an impact on customer satisfaction with airline services , proper handling of luggage and centralized check-in facility (Ringle et al., 2011), enhanced safety measures and onboard entertainment options (Ali et al., 2020), lost baggage and flight cancellations, poor luggage handling, frequency of flights, punctuality of departures and arrivals,,timely delivery of baggage upon arrival, cleanliness of the cabin, quality of food and quantity served during the flight (Jager et al., 2012). (Sukri, 2014)The study found that when there was a service failure, passengers required clear explanations and improved services in order to feel satisfied with the situation. Figure 1. Conceptual Framework of the Relationship between Independent and Dependent Variables 2.4 Hypotheses formulated Hypothesis 1 H0: There is a negative relationship between service failure with passenger satisfaction. Figure 1. Conceptual Framework of the Relationship between Independent and Dependent Variables ceptual Framework of the Relationship between Independent and Dependent Variables 2.4 Hypotheses formulated Hypothesis 1 2.4 Hypotheses formulated Hypothesis 1 yp H0: There is a negative relationship between service failure with passenger satisfaction. yp H0: There is a negative relationship between service failure with passenger satisfaction Volume 6, Issue 2, March-April 2024 Volume 6, Issue 2, March-April 2024 3 ational Journal for Multidisciplinary Research (IJFMR) the Cronbach’s alpha for service failure, service recovery, and passenger satisfaction falls between .7 and .8, which is considered satisfactory and good. Table 2. Major types of service failure (n=361) Table 2. Major types of service failure (n=361) The primary form of service failure encountered by the participants is related to flight issues, accounting for 37.1% of the results, while fares accounted for just 6.1%. The primary form of service failure encountered by the participants is related to flight issues, accounting for 37.1% of the results, while fares accounted for just 6.1%. The primary form of service failure encountered by the participants is related to flight issues, accounting for 37.1% of the results, while fares accounted for just 6.1%. Table 3. Correlation between service failure and passenger satisfaction (n=361) Table 4. Correlation between service recovery and passenger satisfaction (n=361) Table 3. Correlation between service failure and passenger satisfaction (n=361) Table 3. Correlation between service failure and passenger satisfaction (n=361) Table 3. Correlation between service failure and passenger satisfaction (n=361) Table 4. Correlation between service recovery and passenger satisfaction (n=361) Table 4. Correlation between service recovery and passenger satisfaction (n=361) The study's findings are presented in Table 4.3 and 4.4, revealing a significant negative correlation between service failure and passenger satisfaction. Conversely, the relationship between service recovery and passenger satisfaction is positively linked, highly correlated, and significant. 4.1 Survey Return rate Out of 372 surveys handed out, 361 were completed and returned, resulting in a response rate of 97%. Table 1. Reliability Analysis of Variables Table 1. Reliability Analysis of Variables The reliability assessment was created to determine if the components in the survey were categorized under a factor that is dependable and enduring. As per Sekaran, reliabilities below 0.6 are inadequate, those within the 0.7 range are acceptable, and those exceeding 0.8 are excellent. Table 4.1 displays that Volume 6, Issue 2, March-April 2024 IJFMR240217568 4 ational Journal for Multidisciplinary Research (IJFMR) interactions. This methodological choice not only enables the identification of significant predictors but also facilitates a nuanced understanding of their respective contributions in elucidating the multifaceted phenomenon under investigation Steven et al., 2012). Thus, through the meticulous application of multiple regression analysis, the study endeavors to provide valuable insights into the pivotal factors influencing service failure and recovery within passenger relationships, thereby contributing to the broader body of knowledge in the field of service management and consumer behavior. Table 5. Table of Summary and ANOVA (Passenger Satisfaction) Table 5. Table of Summary and ANOVA (Passenger Satisfaction) R Square denotes the proportion of variance in the passenger satisfaction (dependent variable) that is accounted for by the variation in service failure and service recovery (independent variables). With an r square of 0.528, around 52.8% of the variability in passenger satisfaction can be attributed to these variables, while approximately 47.2% remains unexplained. The R value indicates a strong correlation at 72.6%, signifying a significant amount of shared variability between observed and predicted values for passenger satisfaction. The F-value is 200.066 and it is considered significant with a p-value of less than 0.05. If the p-value, denoted as p, exceeds 0.05, it suggests that the independent variables do not significantly explain the dependent variable. Thus, the independent variables of service failure and service recovery have a significant impact on explaining passenger satisfaction as the dependent variable. Additionally, this indicates that the questions are well-suited for analyzing the data. Table 6. Table of Coefficients Table 6. Table of Coefficients Table 4 displays the data regarding the impact of independent variables. The ANOVA table above indicates a significant p-value of .000. Service recovery has the highest beta coefficient at 0.735, making it the most influential variable on the dependent variable. In contrast, service failure has a slightly lower beta value (0.26), indicating less contribution to passenger satisfaction as the dependent variable. Consequently, it can be inferred that while service failure as an independent variable is less impactful, service recovery consistently predicts the dependent variable more effectively. 4.3 Multiple Regression In order to comprehensively examine the intricate interplay between a single continuous dependent variable and multiple independent variables or predictors, the research methodology employed multiple regression analysis. This analytical approach was chosen strategically to fulfill the overarching objective of the study, which aimed to discern the principal predictor or the most influential factor pertaining to service failure and subsequent recovery within the context of passenger relationships (Krejcie & Morgan, 1970). By harnessing the power of multiple regression analysis, the research endeavors to unravel the complex web of relationships between the various independent variables and the dependent variable, thereby shedding light on the key drivers shaping the dynamics of service failure and recovery in passenger Volume 6, Issue 2, March-April 2024 IJFMR240217568 IJFMR240217568 5 6. CONCLUSION Passengers traversing through Sultan Ismail Petra Airport are predisposed to harbor negative perceptions when confronted with service failures, underscoring the pivotal role of effective service recovery mechanisms in assuaging their discontent. It is evident that passengers not only anticipate but also demand heightened efforts and assistance from service providers to rectify any encountered issues, thereby shaping their subsequent levels of satisfaction or disappointment. The intrinsic link between service recovery endeavors and passenger satisfaction highlights the paramount importance for airline companies to deploy robust strategies aimed at ameliorating service failures and fostering enduring customer relationships. Moreover, the findings of this study illuminate flight-related issues as the predominant source of service failures at Sultan Ismail Petra Airport, emphasizing the imperative for airline management to direct attention towards mitigating these recurring challenges. By addressing the root causes of flight-related issues and optimizing the service distribution process, airlines can engender a seamless travel experience for passengers, thereby enhancing overall satisfaction levels and bolstering their competitive edge in the industry landscape. Passengers traversing through Sultan Ismail Petra Airport are predisposed to harbor negative perceptions when confronted with service failures, underscoring the pivotal role of effective service recovery mechanisms in assuaging their discontent. It is evident that passengers not only anticipate but also demand heightened efforts and assistance from service providers to rectify any encountered issues, thereby shaping their subsequent levels of satisfaction or disappointment. The intrinsic link between service recovery endeavors and passenger satisfaction highlights the paramount importance for airline companies to deploy robust strategies aimed at ameliorating service failures and fostering enduring customer relationships. Moreover, the findings of this study illuminate flight-related issues as the predominant source of service failures at Sultan Ismail Petra Airport, emphasizing the imperative for airline management to direct attention towards mitigating these recurring challenges. By addressing the root causes of flight-related issues and optimizing the service distribution process, airlines can engender a seamless travel experience for passengers, thereby enhancing overall satisfaction levels and bolstering their competitive edge in the industry landscape. Furthermore, it is discerned that passengers exhibit a heightened sense of trust and deepen their commitment to the airline when confronted with effective service recovery initiatives from setbacks. Thus, by prioritizing and diligently implementing service recovery strategies, such as timely resolution of issues and proactive communication, AirAsia can cultivate a loyal customer base and fortify its reputation as a customer-centric airline provider. 6. CONCLUSION In essence, the insights gleaned from this study underscore the imperative for airline companies to proactively address service failures, particularly flight-related issues, and invest in robust service recovery mechanisms to nurture enduring customer relationships and sustain competitive advantage in the dynamic aviation sector..” International Journal for Multidisciplinary Research (IJFMR) E-ISSN: 2582-2160 ● Website: www.ijfmr.com ● Email: editor@ijfmr.com service failure encountered by passengers within the airport premises. Furthermore, the findings illuminate a noteworthy finding regarding the relationship between service failure and passenger satisfaction, demonstrating a significant negative correlation between the two variables. Contrary to the initial hypothesis posited, which anticipated a different outcome, the empirical evidence suggests that service failure detrimentally impacts passenger satisfaction levels. However, in line with hypothesis 2, the study corroborates the notion that service recovery exerts a positive influence on passenger satisfaction. This pivotal finding underscores the critical role played by effective service recovery mechanisms in mitigating the adverse effects of service failures and bolstering passenger satisfaction levels. Consequently, it can be inferred that service recovery emerges as the linchpin factor in shaping passenger perceptions and fostering positive experiences within the airline industry. Thus, this study underscores the paramount importance for airlines, including AirAsia, to prioritize and enhance their service recovery strategies as a means to bolster overall passenger satisfaction and loyalty. service failure encountered by passengers within the airport premises. Furthermore, the findings illuminate a noteworthy finding regarding the relationship between service failure and passenger satisfaction, demonstrating a significant negative correlation between the two variables. Contrary to the initial hypothesis posited, which anticipated a different outcome, the empirical evidence suggests that service failure detrimentally impacts passenger satisfaction levels. However, in line with hypothesis 2, the study corroborates the notion that service recovery exerts a positive influence on passenger satisfaction. This pivotal finding underscores the critical role played by effective service recovery mechanisms in mitigating the adverse effects of service failures and bolstering passenger satisfaction levels. Consequently, it can be inferred that service recovery emerges as the linchpin factor in shaping passenger perceptions and fostering positive experiences within the airline industry. Thus, this study underscores the paramount importance for airlines, including AirAsia, to prioritize and enhance their service recovery strategies as a means to bolster overall passenger satisfaction and loyalty. 5. DISCUSSION The primary objective of this study is to delve into the intricate dynamics between service failure and subsequent service recovery in the context of passenger satisfaction, particularly focusing on AirAsia passengers at Sultan Ismail Petra Airport. Through a rigorous analysis of data collected from a sample of AirAsia passengers, this research reveals that flight-related issues emerge as the predominant cause of Volume 6, Issue 2, March-April 2024 IJFMR240217568 6 rnational Journal for Multidisciplinary Research (IJFMR) grievances, airlines can not only salvage potentially compromised customer relationships but also engender a heightened sense of trust and loyalty among their clientele. The findings underscore the critical importance of feedback and complaint management in the airline industry ecosystem. Given the inherently service-oriented nature of the aviation sector, wherein customer experiences play a decisive role in shaping brand perceptions and loyalty, airline companies must establish streamlined processes for capturing, analyzing, and acting upon customer feedback and complaints. By leveraging such insights, airlines can identify recurring issues, pinpoint areas for improvement, and tailor their service offerings to better align with passenger expectations, thereby fostering a culture of continuous improvement and customer-centricity. This study advocates for the exploration of service domains beyond the aviation sector, such as the industrial or hospitality sectors, where service failures are prevalent. By delving into analogous contexts characterized by similar service delivery challenges, researchers can glean valuable insights and best practices that can be extrapolated to inform service management strategies within the airline industry. Additionally, there is a compelling need for future research endeavors to extend beyond domestic airline contexts and encompass international airlines, thereby shedding light on the diverse cultural influences that shape passenger perceptions and expectations across different geographic regions. In essence, the recommendations emanating from this study underscore the imperative for airline companies to adopt a proactive and customer-centric approach towards service recovery and complaint management, while also advocating for broader interdisciplinary research initiatives aimed at advancing our understanding of service failures and recovery mechanisms across diverse service sectors and cultural contexts. By embracing these recommendations and leveraging the insights garnered from interdisciplinary research endeavors, airline companies can position themselves at the forefront of service excellence and forge enduring customer relationships in an increasingly competitive marketplace. 7. RECOMMENDATION Numerous valuable suggestions have surfaced from this study, each offering strategic avenues for airline companies to enhance their customer satisfaction levels and fortify repurchase intentions through the implementation of robust service recovery systems. Recognizing the pivotal role of service recovery in mitigating the adverse effects of service failures on passenger perceptions, it is imperative for airline companies to prioritize the development and deployment of effective mechanisms aimed at promptly addressing and resolving customer grievances. By proactively attending to passenger concerns and Volume 6, Issue 2, March-April 2024 IJFMR240217568 7 References 1. Ali, S R O., Said, N S M., Jislan, F., Mat, K A., & Aznan, W N M W. (2020, April 1). The Relationship between Service Failure and Service Recovery with Airline Passenger Satisfaction. IOP Publishing, 1529(2), 022062-022062. https://doi.org/10.1088/1742-6596/1529/2/022062 2. Bankova. (2018, May 3). Airline Service Failures: A study on relationshipsbetween lack of control, emotions, and negative word-of-mouth.. https://hj.diva- portal.org/smash/record.jsf?pid=diva2%3A1215218&dswid=-1064 3. Chahal, H., & Devi, P. (2015, February 2). 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https://openalex.org/W2577578864	OpenAlex	Open Science	CC-By	2,017	Assessing Sexual Dicromatism: The Importance of Proper Parameterization in Tetrachromatic Visual Models	Pierre‐Paul Bitton	English	Spoken	14,027	23,492	Assessing Sexual Dicromatism: The Importance of Proper Assessing Sexual Dicromatism: The Importance of Proper Parameterization in Tetrachromatic Visual Models Parameterization in Tetrachromatic Visual Models Pierre-Paul Bitton University of Windsor Pierre-Paul Bitton University of Windsor Kevyn Janisse Stéphanie M. Doucet Biological Sciences, University of Windsor Stéphanie M. Doucet Biological Sciences, University of Windsor Follow this and additional works at: https://scholar.uwindsor.ca/biologypub Part of the Biology Commons Part of the Biology Commons University of Windsor University of Windsor Scholarship at UWindsor Scholarship at UWindsor Biological Sciences Publications Department of Biological S 1-11-2017 Assessing Sexual Dicromatism: The Importance of Proper Assessing Sexual Dicromatism: The Importance of Proper Parameterization in Tetrachromatic Visual Models Parameterization in Tetrachromatic Visual Models Pierre-Paul Bitton University of Windsor Kevyn Janisse Stéphanie M. Doucet Biological Sciences, University of Windsor Department of Biological Sciences Department of Biological Sciences Pierre-Paul Bitton¤, Kevyn Janisse, Ste´phanie M. Doucet Department of Biological Sciences, University of Windsor, Windsor, Ontario, Canada Pierre-Paul Bitton¤, Kevyn Janisse, Ste´phanie M. Doucet Department of Biological Sciences, University of Windsor, Windsor, Ontario, Canada ¤ Current address: Institute of Evolution and Ecology, Eberhard Karls Universita¨t Tu¨bingen, Tu¨bingen, Germany * pierre-paul.bitton@uni-tuebingen.de ¤ Current address: Institute of Evolution and Ecology, Eberhard Karls Universita¨t Tu¨bingen, Tu¨bingen, Germany * pierre pa l bitton@ ni t ebingen de ¤ Current address: Institute of Evolution and Ecology, Eberhard Karls Universita¨t Tu¨bingen, Tu¨bingen, Germany * pierre-paul.bitton@uni-tuebingen.de a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS OPEN ACCESS Citation: Bitton P-P, Janisse K, Doucet SM (2017) Assessing Sexual Dicromatism: The Importance of Proper Parameterization in Tetrachromatic Visual Models. PLoS ONE 12(1): e0169810. doi:10.1371/ journal.pone.0169810 Recommended Citation Recommended Citation Bitton, Pierre-Paul; Janisse, Kevyn; and Doucet, Stéphanie M., "Assessing Sexual Dicromatism: The Importance of Proper Parameterization in Tetrachromatic Visual Models" (2017). PLoS One, 12, 1, e0169810. https://scholar.uwindsor.ca/biologypub/126 This Article is brought to you for free and open access by the Department of Biological Sciences at Scholarship at UWindsor. It has been accepted for inclusion in Biological Sciences Publications by an authorized administrator of Scholarship at UWindsor. For more information, please contact scholarship@uwindsor.ca. RESEARCH ARTICLE Abstract Perceptual models of animal vision have greatly contributed to our understanding of animal- animal and plant-animal communication. The receptor-noise model of color contrasts has been central to this research as it quantifies the difference between two colors for any visual system of interest. However, if the properties of the visual system are unknown, assump- tions regarding parameter values must be made, generally with unknown consequences. In this study, we conduct a sensitivity analysis of the receptor-noise model using avian visual system parameters to systematically investigate the influence of variation in light environ- ment, photoreceptor sensitivities, photoreceptor densities, and light transmission properties of the ocular media and the oil droplets. We calculated the chromatic contrast of 15 plumage patches to quantify a dichromatism score for 70 species of Galliformes, a group of birds that display a wide range of sexual dimorphism. We found that the photoreceptor densities and the wavelength of maximum sensitivity of the short-wavelength-sensitive photoreceptor 1 (SWS1) can change dichromatism scores by 50% to 100%. In contrast, the light environ- ment, transmission properties of the oil droplets, transmission properties of the ocular media, and the peak sensitivities of the cone photoreceptors had a smaller impact on the scores. By investigating the effect of varying two or more parameters simultaneously, we further demonstrate that improper parameterization could lead to differences between cal- culated and actual contrasts of more than 650%. Our findings demonstrate that improper parameterization of tetrachromatic visual models can have very large effects on measures of dichromatism scores, potentially leading to erroneous inferences. We urge more com- plete characterization of avian retinal properties and recommend that researchers either determine whether their species of interest possess an ultraviolet or near-ultraviolet sensi- tive SWS1 photoreceptor, or present models for both. Introduction association with the American Museum of Natural History to K.J. association with the American Museum of Natural History to K.J. The study of animal visual systems has greatly enhanced our understanding of visual ecology and visual communication. Modelling the sensory perception of various taxa has permitted the study of animal-animal interactions such as mate choice among color morphs in butterflies [1], the evolutionary trade-off between predator-driven crypsis and sexually-selected conspicu- ousness in Dendrobates frogs [2], the influence of insect warning coloration on the predatory behavior of foraging birds [3], and the rejection of brood parasite eggs by host species [4, 5]. Furthermore, visual modelling has been useful in studies of plant-animal interactions. These include, for example, the evolution of flower colors driven by pollinator visual systems [6], the evolution of seed color as a form of crypsis against foraging birds [7], crypsis in plants to avoid predatory herbivores [8], the ability of birds to detect and select high-lipid fruits [9, 10], and the comparative ability of dichromat and trichromat primates in discriminating fruit from leaves [11]. Central to these studies is the concept of color discrimination thresholds limited by photoreceptor noise [12, 13]. This psychophysiological model of chromatic vision quanti- fies perception in color differences [14–16], with the caveat that the light environment and the properties of the visual system, which are central to the model, are well understood. Competing Interests: The authors have declared that no competing interests exist. Visual system properties needed to produce informative visual models must include the photoreceptor sensitivities, transmission properties of the ocular media, the relative photore- ceptor noise levels (approximated by relative photoreceptor densities), and for animals such as birds, lizards, and turtles, the properties of the photoreceptor-specific oil droplets which act as filters and micro-lenses [17–20]. While molecular methods and microspectrophotometry are increasingly used to determine the physical properties of visual systems (e.g., [21, 22]), com- plete characterizations are available for relatively few species. Furthermore, groups of closely related species have rarely been compared (see [23] for an exception). To circumvent these lack of data, research using birds as models have relied on ‘average’ visual system information (calculations and data for ultraviolet sensitive and ultraviolet insensitive eye types presented in [24]), or used parameters from closely related species. Parameterization of Avian Visual Models Editor: Daniel Osorio, University of Sussex, UNITED KINGDOM Received: August 29, 2016 Accepted: December 21, 2016 Published: January 11, 2017 Copyright: © 2017 Bitton et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: Funding was provided by the Natural Sciences and Engineering Research Council of Canada in the form of a Canada Graduate Scholarship to P-P.B. as well as Discovery and Equipment grants to S.M.D, an Ontario Graduate Scholarship to P-P.B., as well as a Field Museum Visiting Scholar Grant and a Collections Study Grant from the Richard Gilder Graduate School in 1 / 22 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Introduction Initial comparative analyses assumed a strong association between visual systems and phylogeny [25–27], but recent studies have shown that this is not always the case. Changes between ultraviolet sensitive (UVS) and violet sensitive (VS) eye types have occurred several times in some Orders (e.g., Passeriformes and Charadriiformes: [28]), and both UVS and VS eye types can be present within the same family (e.g., Maluridae: [29]). Furthermore, the very basic organizations of visual systems can differ among and within Orders. For example, a large majority of birds characterized to date possess four distinctive single cone types (Short-wavelength-sensitive 1 –SWS1, Short-wavelength-sen- sitive 2 –SWS2, Medium-wavelength-sensitive–MWS, and Long-wavelength-sensitive–LWS, (e.g., [30, 31]), but exceptions have been found. The tawny owl (Strix aluco Linnaeus), and pos- sibly other nocturnal raptors, lacks the UVS–VS SWS1 pigment and is therefore trichromatic [32, 33]. In contrast, the bobolink (Dolichonyx oryzivorus Linnaeus; Passeriformes) possesses five distinct classes of single cones: four narrowly tuned photoreceptors and one broadband photoreceptor [34]. Clearly, not all birds share the same visual sensory experience, and the incorrect parameterization of visual models could potentially lead to erroneous conclusions for a variety of ecological questions (see [27, 35, 36] for a debate example). The possible effects of differential visual model parameterization have been explored in a diversity of taxa. Studies comparing dicromats to trichromats [37, 38] and trichromats to tetra- chromats [39, 40] have quantified how these visual systems differ in their ability to tell apart patches with different spectral properties. Others have tested the effect of different light envi- ronments [22, 41–43], photoreceptors sensitivities [22, 44], photoreceptor densities [44, 45], oil droplet characteristics [44, 46–48], ocular media [22], and receptor signal-to-noise ratio PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 2 / 22 Parameterization of Avian Visual Models [44]. While these studies have been very informative when considered together, the use of different visual system starting points and non-standardized methods of presenting results have made it difficult to compare the relative effect of each parameter within a single con- text. In this study, we conduct a sensitivity analysis of the receptor-noise model by systemat- ically testing the effect of single parameters on measures of sexual dichromatism among 70 species of Galliformes, a group characterized by extreme variation in sexual dimorphism. Furthermore, we explore the effects of also varying two or more parameters on three exam- ple species. Introduction The receptor-noise model for color discrimination is commonly used to evaluate sexual dichromatism (differences in coloration) within species, and to characterize color diver- gence among closely related or incipient species (e.g., [49–51]). Sexual dichromatism scores (the sum or average of differences between corresponding male and female plumage patch colors) have been used to study the evolution of sexual dichromatism [52], the influence of sexual selection on sexual dichromatism [53, 54], the relationship between sexual dichroma- tism and conspicuousness [55], and factors that may account for congeneric color diversity [29]. In addition, sexual dichromatism has been used as a proxy for the intensity of sexual selection in comparative studies [54, 56]. Although it has been demonstrated that human visual assessments produce different but similar approximations of dichromatism scores compared to tetrachromatic birds [13, 40, 57, 58], the effects of parameterization of bird visual models in assessments of dichromatism scores has never been systematically deter- mined. In this study, we calculated the chromatic sexual dichromatism scores of 15 color patches for each of 70 species of Galliformes using the receptor-noise model developed by Vorobyev and Osorio [12]. For each patch, sexual dichromatism is calculated as the just- noticeable-difference (JND) in color between males and females. We evaluated the influ- ence of light environments, photoreceptor sensitivities, oil droplet characteristics, ocular transmission, and photoreceptor densities on chromatic contrast values. The purpose of our study was to understand the relative effect of each model parameter on overall dichroma- tism scores, and to guide researchers when making assumptions about visual systems in studies using visual models. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Materials and Methods Species in the Order Galliformes exhibit a tremendous diversity in sexual dichromatism, rang- ing from completely monochromatic to highly dichromatic. Across the various species in this order, feather coloration is predominantly produced by melanin pigmentation or structural colors [59], with very few plumage patches colored by carotenoid pigments [60]. We selected 70 species, most of them broadly distributed across the family Phasianidae (65 of 70 species), and measured 15 plumage patches on three males and three females of each species when available (list of species and specimen museum accession numbers in Table A in S3 File). We obtained spectral reflectance measurements using a USB 4000 spectrophotometer combined with a PX-2 Xenon light source (Ocean Optics, Dunedin, FL). We collected measurements using a bifurcated probe with a rubber stopper tip, which blocked out ambient light and main- tained the probe at normal incidence and ~3 mm above the feather surface. We measured each region five times, haphazardly relocating the probe each time, and used the average of the 15 measurements (five in each of three individuals) in subsequent analyses [61]. The range of col- ors across the species measured covered ~58% of the gamut of structural bird colors obtained in a comprehensive survey of plumage coloration (Fig 1; [62]). PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 3 / 22 Parameterization of Avian Visual Models Fig 1. Molleweide projection of the 2100 color patches used in this study when perceived by an average UVS visual system under ideal illumination. The colors of the symbols are approximations of the colors of the patches based on a human visual system. SWS1, SWS2, MWS, and LWS refer to the ultraviolet-, short-, medium-, and long- wavelength photoreceptor, respectively. d i 10 1371/j l 0169810 001 Fig 1. Molleweide projection of the 2100 color patches used in this study when perceived by an average UVS visual system under ideal illumination. The colors of the symbols are approximations of the colors of the patches based on a human visual system. SWS1, SWS2, MWS, and LWS refer to the ultraviolet-, short-, medium-, and long- wavelength photoreceptor, respectively. doi:10 1371/journal pone 0169810 g001 doi:10.1371/journal.pone.0169810.g001 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models then explored the multidimensional parameter space by calculating the average dichromatism scores generated by every possible combination of parameters presented in this study. We thoroughly surveyed the literature with ISI Web of Science (accessed Dec 1st 2014) for all bird studies reporting quantitatively assessed visual system parameters (e.g., using micro- spectrophotometry), but did not use predicted values based on gene expression (e.g., SWS1 peak sensitivity based on opsin amino acid substitution, [65]). We compiled the information available on avian photoreceptor sensitivities (Table B in S3 File), oil droplet characteristics (Table C in S3 File), transmission properties of ocular media (Table C in S3 File), and photore- ceptor retinal densities (Table D in S3 File). We summarized these data separating UVS from VS eye type. We also extracted from the literature the most commonly used light environ- ments (see below for more details). We tested the influence of each visual model parameter by comparing dichromatism scores obtained from systematically changing the value of a single parameter. This was accomplished using the R package pavo functions (sensmodel, vismodel, and coldist; [66, 67]). Within-param- eter effects were evaluated first by comparing each set of dichromatism scores against those obtained using the most commonly implemented visual models, the average VS or UVS eye type [24], followed by pairwise comparisons to find the parameter values that produced the most divergent scores. The effects of parameterization were assessed in two ways: 1) by calculat- ing the Pearson’s correlation coefficient between the mean dichromatism scores, and 2) by com- paring the ranks of the mean dichromatism scores (as per [40]). We determined how many species maintained the same rank, the mean and standard deviation of the change in rank, and the maximum change in rank. The species most affected by the changes in parameterization (largest absolute difference in mean dichromatism score and largest change in rank) were selected for further analysis. For these species, we determined the number of patches (out of 15) considered non-distinguishable under each of the two visual models, the number of patches that changed by more than 1 JND, and the maximum dichromatism score change in a single patch (in JNDs and percentage). Endler and Mielke (2005) indicate a value of 362 nm for the ocular media cut-off point but we could only reproduce the photoreceptor curves from their supplemental material when using 348 nm. These visual systems were used as the starting point for comparison of the various parameters. λ cut: Oil droplet cut-off wavelength; Bmid: Gradient of line tangent to absorbance spectrum at λ mid. S1 photoreceptor do not ﬁlter light between 300 and 700 nm. General procedures We tested the general influence of visual model parameterization on the mean dichromatism score of each of the 70 species, and further examined the species most affected by the different parameters. For each plumage patch, we compared the spectral reflectance of the male color to that of the female color by calculating the chromatic contrast (the difference between the col- ors in JNDs). We calculated the mean dichromatism score for each species as the sum of dichromatism scores across all patches divided by the number of patches (e.g., [52, 63, 64]). Some species possess non-feathered facial patches which, in museum skins, loose their color. For these species, the mean score was calculated based on the number of feathered patches. JND values smaller than 1 (i.e., non-discriminable) were given values of ‘0’ to avoid inflating the dichromatism scores based on non-detectable differences. For three of the 70 species, we PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 4 / 22 doi:10.1371/journal.pone.0169810.t001 Model parameterization Therefore, we evaluated the influence of changes in single photoreceptor peak sensitivities, using the mini- mum and maximum reported for each photoreceptor type for each eye type (Table E in S3 File), and then compared visual models that expressed either all minimum or all maximum peak sensitivity values. Oil droplets–In birds, each photoreceptor type is paired with a specific oil droplet type that acts as a cut-off filter and microlens [13, 17, 18]. SWS1 photopigments are associated with mostly transparent droplets (T type), SWS2 pigments with droplets clear in appearance (C type), MWS pigments with yellow droplets (Y type), and LWS pigments with red droplets (R types). The absorption profile of the oil droplets can be extrapolated if the wavelength at which the oil droplet transmittance equals 1/e (λo) and the absorptivity rate of decay (b) are known. In turn, these properties can be estimated from the cut-off wavelength (λ cut) and the gradient of line tangent to the absorbance spectrum (Bmid) at the wavelength at half-maximum absor- bance (λ mid), the only values that are reported in some studies of oil droplet characteristics (formulae presented in [17] and reproduced in S2 File with acronyms in S1 File). Because there is no strong relationship between a visual pigment’s peak sensitivity and the absorbance characteristics of its oil droplet (within droplet type) [17], we evaluated the influence of differ- ences in extreme cut-off values within photopigment type first by changing single oil droplet parameters, and then by comparing visual systems with oil droplet values set with all maxi- mums and all minimums, by eye type (Table F in S3 File). Extremes were selected based on λ cut, and actual values of either Bmid (if available from the literature) or Bmid calculated from λ mid (Table C in S3 File). Ocular media–Similar to oil droplets, the cornea and lens act as a cut-off filters. Recent work has demonstrated that phylogeny and eye type can be used to estimate the approximate high-pass cut-off values of the ocular media in birds [22], but that variability within UVS and VS eye types, and in certain groups, is very high (e.g., waterbirds [21]). The absorption curves of ocular media in birds are all very similar and can be well approximated (function from [24] reproduced in S2 File) when the wavelength at 50% transmission (T50) is known. Model parameterization Model parameterization Average visual model–Our basis for comparisons were the two average visual models (UVS and VS eye type) presented by Endler and Mielke [24] (Table 1). In addition to the parameter Table 1. Parameters used to reproduce the average VS and average UVS avian visual systems presented in Endler and Mielke (2005). Eye type Ocular media (nm) Parameter UVS/VS SWS MWS LWS UVS 324 Peak sensitivities (nm) 367 444 501 564 λ cut NA† 411 511 572 Bmid NA 0.0278 0.023 0.022 VS 348 Peak sensitivities (nm) 412 452 505 565 λ cut NA 447 510 570 Bmid NA 0.0294 0.028 0.020 arameters used to reproduce the average VS and average UVS avian visual systems presented in Endler and rage VS and average UVS avian visual systems presented in Endler and Mielke (2005). Table 1. Parameters used to reproduce the average VS and average UVS avian visual systems presente These visual systems were used as the starting point for comparison of the various parameters. λ cut: Oil droplet cut-off wavelength; Bmid: Gradient of line tangent to absorbance spectrum at λ mid. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 5 / 22 Parameterization of Avian Visual Models values detailed in Table 1, we used the relative photoreceptor densities of the Pekin Robin (Leiothrix lutea Scopoli; SWS1:SWS2:MWS:LWS = 1:2:2:4) as used originally by Vorobyev and Osorio [12] and Vorobyev et al. [13], and set the Weber fraction to 0.05 for the LWS pho- toreceptor ([13, 68, 69]; Formulae detailed in S2 File). Light environment–We compared the influence of the seven most commonly used environ- mental illuminants [70, 71]: 1) Ideal (wavelength independent), 2) Forest Shade, 3) Woodland shade, 4) Blue sky, 5) daylight D65 standard [72], 6) Woodland Gaps, and 7) Cloudy sky. Photoreceptor sensitivities–The large majority of avian species possess four retinal cone pigments: two short-wavelength- sensitive pigments SWS1 and SWS2, one medium-wave- length-sensitive pigment MWS, and one long-wavelength-sensitive pigment LWS. The spec- tral sensitivity curves of these pigments can be accurately estimated from the wavelength of maximum sensitivity using a near-universal template [73]. The peak wavelength sensitivities of the four avian photoreceptors among species are correlated [17], but still allow for consider- able variation in the individual sensitivity values within any given visual system. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Model parameterization We evalu- ated the influence of varying the T50 value between 314 nm and 344 nm for UVS type eyes (the range of values known to exist for this eye type, Table C in S3 File), and between 335 nm and 395 nm in VS eye type (Table C in S3 File), using four values with equal intervals (10 nm increments among models for the UVS eye type and 20 nm for the VS eye type, Table F in S3 File). Photoreceptor densities–The relative densities of photoreceptors vary within and among species, and even within individuals, with some evidence for bilateral asymmetry in at least two species (European starling Sturnus vulgaris Linnaeus and Blue tit Cyanistes caeruleus PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 6 / 22 Parameterization of Avian Visual Models Linnaeus [31, 74]). Furthermore, photoreceptor densities are more heavily influenced by the ecology of the species (diet, feeding behavior, habitat) rather than phylogeny [31]. Therefore, patterns of receptor densities are difficult to predict. We tested the influence of this parameter by selecting nine different photoreceptor densities reported in a variety of species (not includ- ing the original 1:2:2:4; see Table G in S3 File for species and reasoning behind inclusion). So that the models would be comparable, we maintained the Weber fraction of the most abundant photoreceptor at 0.05 by using a different standard deviation of noise in a receptor for each model [13, 75]. Model visual systems–The physical properties of visual systems have been completely char- acterized in only eight species (See Results). These systems were compared to each other and to either the average UVS or VS system based on the peak wavelength sensitivity of their SWS1 photoreceptor. Because they are the most commonly used sets of parameter values, we also compared the dichromatism scores generated using the average UVS and VS eye type visual models. For all models we used a Weber fraction of 0.05 (the most commonly used), which was empirically determined for the LWS photoreceptor of Leiothrix lutea [68], and confirmed though behavioral tests in domesticated chicken Gallus gallus Linnaeus [69]. Multidimensional parameter space–To explore the effects of modifying one or more than one parameters at the same time, we selected three of the 70 species (Tetraogallus tibetanus, Arborophila torqueola, and Lophophorus impejanus) and calculated their average dichromatism scores for all possible combinations of parameter conditions detailed above. Model parameterization We selected these species using three criteria: 1) perceived by an average visual system of a VS eye-type, these species’ dichromatism score represent a low score species, an average score species, and a high score species, 2) they do not possess any bare parts, and 3) some of the patches (using the aver- age visual system of a VS eye type) have values below 1 JND. For each eye type, the parameters comprised seven light environments including the commonly used ‘ideal’ illuminant, four T50 ocular media values, 11 combinations of spectral sensitivity values including the ‘average’ pas- serine, nine combinations of photoreceptor oil droplet values including the ‘average’ passerine, and nine photoreceptor density ratios including the ‘average’ passerine. We generated the mean dichromatism scores for all 24 948 possible permutations for each eye type, and present descriptive statistics for mean scores and the number of patches with scores above 1 JND. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models ons that generated the largest differences, based on the lowest Pearson’s r value, in the mean alliformes. Table 2. Summary of the visual system comparisons that generated the largest differences, based on the lowest Pearson’s r value, in the mean dichromatism scores of 70 species of the Order Galliformes. Parameter Eye type Conditions Pearson’s r Equal rank Rank change Rank SD Max change Light environment UVS Ideal vs D65 0.9991 30 1.03 1.57 10 VS Ideal vs D65 0.9994 36 0.77 1.23 8 Photoreceptor λmax UVS All Max vs All Min 0.9986 27 1.03 1.20 5 VS All Max vs All Min 0.9963 17 1.54 1.71 9 Oil droplet cut-off value UVS R Min vs All Max 0.9979 27 1.00 1.13 6 VS R Max vs All Min 0.9947 21 1.26 1.20 5 Ocular media T50 values UVS T314 vs T344 0.9998 45 0.51 0.85 4 VS T335 vs T395 0.9991 28 1.06 1.46 9 Photoreceptor densities UVS A. minutus vs P. paciﬁcus 0.9791 8 3.11 2.87 12 VS A. minutus vs P. paciﬁcus 0.9846 6 2.63 2.07 10 Model systems - Average UVS vs Average VS 0.9929 15 2.34 2.66 16 - P. cristatus vs S. vulgaris 0.9881 8 3.00 3.06 16 Values reported describe how many species (out of 70) were assigned the same rank in the comparison (Equal rank), the average rank change (Rank change), the standard deviation of rank change (Rank SD), and the maximum rank change (Max change). See Materials and Methods, Results, and Supporting Information sections for more details. Table 2. Summary of the visual system comparisons that generated the largest differences, based on the lowest Pearson’s r value, in the mean dichromatism scores of 70 species of the Order Galliformes. Values reported describe how many species (out of 70) were assigned the same rank in the comparison (Equal rank), the average rank change (Rank change), the standard deviation of rank change (Rank SD), and the maximum rank change (Max change). See Materials and Methods, Results, and Supporting Information sections for more details. doi:10.1371/journal.pone.0169810.t002 the largest influence on the mean dichromatism scores and rank differences. Variation in the transmission properties of the ocular media (within eye type) and the light environment had less influence on the mean dichromatism scores and on the ranks of species (Table 2). Results The mean dichromatism scores across the 70 species were always highly correlated regardless of which visual system parameter was altered (Summary in Table 2; details in Tables H–M in S3 File). Pearson’s coefficient (r) of the largest differences, within parameter, ranged from 0.9998 when contrasting the extreme T50 ocular media values for the UVS eye type (Table 2, Fig 2A), to 0.9791 when contrasting the photoreceptor densities of Anous minutus Boie (Black noddy) to those of Puffinus pacificus Gmelin (Wedged-tailed shearwater) in a UVS eye type (Table 2, Fig 3A). However, there was considerable variation in the number of species that maintained the same dichromatism rank, the mean rank change, and the maximum rank change. For example, the extreme T50 ocular media values for the UVS eye type had a moder- ate effect on the overall ranks and rank changes (45 out of 70 with equal rank, a mean rank change of 0.51, and maximum rank change of 4; Fig 2B), but the differences in photoreceptor densities in the UVS eye type had a large impact on the mean dichromatism ranks (only 8 out of 70 with equal rank, a mean rank change of 3.11, and maximum rank change of 12; Fig 3B). Comparisons of the dichromatism scores calculated with the commonly-used average UVS and average VS eye sets of conditions produced one of the largest difference in rank scores (Table 2, Fig 4B). Overall, photoreceptor densities and the variation among model systems had PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 7 / 22 Values reported describe how many species (out of 70) were assigned the same rank in the comparison (Equal rank), the average rank change (Rank change), the standard deviation of rank change (Rank SD), and the maximum rank change (Max change). See Materials and Methods, Results, and Supporting Information sections for more details. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 doi:10.1371/journal.pone.0169810.t002 Parameterization of Avian Visual Models Fig 2. Effect of the ocular media parameter on calculated sexual dichromatism scores. Comparison of the A) mean sexual dichromatism scores (in JNDs), and B) sexual dichromatism ranks of 70 galliform species for a UVS eye type with different ocular media transmission. Values on the x-axis were generated by setting the T50 of the ocular media at 314 nm (see formulae in S2 File) and the values on the y-axis were generated by setting the T50 of the ocular media at 344 nm. The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. doi 10 1371/jo rnal pone 0169810 g002 Fig 2. Effect of the ocular media parameter on calculated sexual dichromatism scores. Comparison of the A) mean sexual dichromatism scores (in JNDs), and B) sexual dichromatism ranks of 70 galliform species for a UVS eye type with different ocular media transmission. Values on the x-axis were generated by setting the T50 of the ocular media at 314 nm (see formulae in S2 File) and the values on the y-axis were generated by setting the T50 of the ocular media at 344 nm. The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Analyzing the scores of individual species most affected by changes in visual model values, based on the largest differences in mean dichromatism scores, we found that large changes in plumage patch chromatic contrast (in JND) can occur when manipulating single parameters (Table 3). The maximum changes in percentage JNDs ranged from 12.5% (0.99 JNDs) when comparing the extreme T50 ocular media values for the UVS eye type, to 84.16% (5.82 JNDs) when comparing an UVS eye type to a VS eye type (Fig 4C). Different model systems also had large effects on the chromatic contrast of some patches (Table 3). The species most affected in their mean dichromatism scores almost always had the same number of non-dichromatic patches under the two sets of conditions (comparing column 1 and column 2 of Table 3), but the number of patches that changed by more than 1 JND (column 3 Table 3) varied consider- ably across parameters. For the T50 ocular media value in a UVS eye, none of the patches changed by at least 1 JND. In contrast, 14 patches changed by more than 1 JND when compar- ing the photoreceptor densities of A. minutus and P. pacificus using the UVS eye type. Overall, individual patches were mostly influenced by condition changes in the photoreceptor densities and the variation among model systems. We also found large differences in patch dichroma- tism scores when comparing the average UVS and average VS eye type (Fig 4C). Variation in light environments, photoreceptor sensitivities, and ocular media values had relatively small but notable effects on the dichromatism scores of some patches (Table 3). When analyzing the scores of individual species most affected by changes in visual model values, based on the largest differences in dichromatism ranks, we found that large differences in ranks were associated with changes in the number of distinguishable patches (number of patches with 0 JNDs) under the two conditions (Table 4). The parameter conditions that gen- erated the largest changes in ranks also had large differences in the number of distinguishable patches under the different sets of parameters. For example, the extreme T50 ocular media PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 8 / 22 doi:10.1371/journal.pone.0169810.g002 values for the UVS eye type differed by only a single patch that changed by more than 1 JND (4 patches with JND < 1 for T50 of 314 nm compared to 5 for T50 of 344 nm). In contrast, comparison of the A. minutus (8 patches with JND < 1) and P. pacificus (1 patch with JND < 1) photoreceptor densities in a UVS eye type generated 12 patches with changes in JND > 1 (Table 4). Contrary to species most affected when comparing mean dichromatism scores, we did not find large changes in plumage patch chromatic contrast (in JND) when manipulating single parameters. Indeed, none of the patches differed by more than 3.6 JNDs across all parameter values. Differences in light environment, photoreceptor sensitivities, and ocular media values generally had the smallest effects on dichromatism scores of individual patches in species with the greatest change in dichromatism ranks; changes in photoreceptor densities, and comparisons of model systems generated the largest effects (Table 4). PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 9 / 22 Parameterization of Avian Visual Models Fig 3. Effect of the photoreceptor density parameter on calculated sexual dichromatism scores. Comparison of A) the mean sexual dichromatism scores (in JNDs), and B) ranks of 70 galliform species for a UVS eye type with different photoreceptor densities. Values on the x- axis were generated using the parameters associated with the ‘average UVS eye-type’ visual system but with the photoreceptor densities found in Anous minutus (SWS1–1.00, SWS2–9.59, MWS– 16.82, LWS– 14.29); values on the y-axis were generated with the ‘average UVS eye- type’ visual system but with the photoreceptor densities found in Puffinus pacificus (SWS1–1.00, SWS2–0.68, MWS– 1.04, LWS– 1.44; see Methods). The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. Fig 3. Effect of the photoreceptor density parameter on calculated sexual dichromatism scores. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models Fig 4. Effect of using either the UVS or VS average eye-type on calculated sexual dichromatism scores. Comparison of A) the mean sexual dichromatism scores (in JNDs), and B) ranks of 70 galliform species contrasting the two most commonly used bird visual systems. Values on the x-axis were generated using the parameters associated with the ‘average UVS eye-type’ visual system; values on the y-axis were generated with the ‘average VS eye-type’ visual system (see Methods). The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. Fig 4. Effect of using either the UVS or VS average eye-type on calculated sexual dichromatism scores. Comparison of A) the mean sexual dichromatism scores (in JNDs), and B) ranks of 70 galliform species contrasting the two most commonly used bird visual systems. Values on the x-axis were generated using the parameters associated with the ‘average UVS eye-type’ visual system; values on the y-axis were generated with the ‘average VS eye-type’ visual system (see Methods). The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. doi:10.1371/journal.pone.0169810.g004 doi:10.1371/journal.pone.0169810.g004 range, substantially more than T. tibetanus, with the mean score of the UVS eye type being on average 1.08 times greater the mean VS eye type scores. doi:10.1371/journal.pone.0169810.g004 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Comparison of A) the mean sexual dichromatism scores (in JNDs), and B) ranks of 70 galliform species for a UVS eye type with different photoreceptor densities. Values on the x- axis were generated using the parameters associated with the ‘average UVS eye-type’ visual system but with the photoreceptor densities found in Anous minutus (SWS1–1.00, SWS2–9.59, MWS– 16.82, LWS– 14.29); values on the y-axis were generated with the ‘average UVS eye- type’ visual system but with the photoreceptor densities found in Puffinus pacificus (SWS1–1.00, SWS2–0.68, MWS– 1.04, LWS– 1.44; see Methods). The solid triangle symbol in A) identifies the species that experienced the greatest change in mean dichromatism score, and the greatest change in rank in B). The sexual dichromatism score of each patch of the species highlighted in A), under the two sets of parameters, are presented in C). The sexual dichromatism score of each patch of the species highlighted in B), under the two sets of parameters, are presented in D). In C) and D), the solid triangle symbol identifies the patch that experienced the greatest change in dichromatism score. Dashed lines represent 1:1 reference line. doi:10.1371/journal.pone.0169810.g003 doi:10.1371/journal.pone.0169810.g003 The permutation of all conditions included in the sensitivity analyses presented above gen- erated 24 948 dichromatism scores per species per eye type (Fig 5). For Tetraogallus tibetanus, a species with low dichromatism scores, the mean scores ranged from 0.41 to 2.81 JNDs when combining the UVS and VS permutation scores, a 685% difference between the lowest and highest values (Table N in S3 File). The distribution of scores for the two eye types overlapped over only 58% of their combined range, with the mean score of the UVS eye type being on average 1.51 times greater the mean VS eye type scores. For Lophophorus impejanus, a species with high dichromatism scores, the mean scores ranged from 5.26 to 14.77 JNDs when com- bining the UVS and VS permutation scores, a 281% difference between the lowest and highest values. The distribution of scores for the two eye types overlapped over 80% of their combined PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 10 / 22 Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. doi:10.1371/journal.pone.0169810.t003 varied little for A. torqueola and L. impejanus, with a large majority of permutations indicating 13 and 15 patches, respectively, with values above 1 JND. For Arborophila torqueola, a species with mid-range dichromatism scores, the mean scores ranged from 3.00 to 7.68 JNDs when combining the UVS and VS permutation scores, a 256% difference between the lowest and highest values. The distribution of scores for the two eye types overlapped over 70% of their combined range, with the mean score of the UVS eye type being on average 1.11 times greater than the mean VS eye type scores. In accordance with results obtained when modifying single parameters, the species with the lowest mean dichromatism score, T. tibetanus in this case, var- ied most in the number of patches with scores above 1 JND across all permutations (Fig 5). The number of patches above discrimination threshold ranged from four to 10 in VS eye type, and from seven to 10 in UVS eye type. The number of patches above discrimination threshold PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 11 / 22 Parameterization of Avian Visual Models omatism score of the species most affected by changes in the sensory exprience, based on the absolute core Table 3. Summary of the changes in dichromatism score of the species most affected by changes in the sensory exprience, based on the absolute largest difference in mean dichromatism score. Parameter Eye type Conditions Condition 1 (< 1 JND) Condition 2 (< 1 JND) > 1 JND change Max change (JNDs) Max change (%) Light environment UVS Ideal vs D65 4 4 4 1.30 25.95 VS Ideal vs D65 10 14 4 1.22 NA Photoreceptor λmax UVS All Max vs All Min 4 4 5 1.34 22.95 VS All Max vs All Min 3 3 5 2.75 17.77 Oil droplet cut-off value UVS R Min vs All Max 1 1 9 4.73 28.53 VS R Max vs All Min 1 1 10 7.16 41.64 Ocular media T50 values UVS T314 vs T344 0 0 0 0.99 12.48 VS T335 vs T395 9 14 5 1.33 NA Photoreceptor densities UVS A. minutus vs P. paciﬁcus 1 1 14 7.16 54.13 VS A. minutus vs P. paciﬁcus 3 3 10 10.47 49.19 Model systems - Average UVS vs Average VS 0 0 9 5.82 84.16 - P. cristatus vs S. vulgaris 1 1 11 8.43 32.12 Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. doi:10.1371/journal.pone.0169810.t004 differences among the models, but found that the rank of individual species could be greatly affected. These results imply that even though parameterization has little effect on the general distribution of the mean dichromatism scores, changes in visual system assumptions can have large effects on the relative position of the mean dichromatism scores of species in relation to one another. Such variation could be important when comparing dichromatism scores to other ecological or life history traits. Furthermore, certain parameters had a large influence on the number of patches considered above discrimination threshold (JND > 1), and the dichro- matism scores of individual patches. Consequences of incorrect parameterization could easily lead to the improper inference that species are sexually dichromatic or not. Our findings sug- gest that the parameterization of avian visual systems should not be trivialized. Indeed, the dif- ferences in chromatic contrast we documented between patches were in some cases as large as those reported when comparing color discrimination between trichromats and tetrachromats [40]. Discussion Compared to fish and invertebrates, avian visual systems exhibit considerably less variation in physical and physiological properties [71]. Nevertheless, a number of studies suggest that vari- ation in avian visual systems appears to be adaptive. Carotenoid-based signals are aligned with cone sensitivities across species of the Passerida clade of passerine birds [76]; the expression of opsin genes are associated with sexual dichromatism in New World warblers [77]; the ocular media of UVS eye types allow more UV light to reach the retina than the ocular media of VS eye types [78]; and the photoreceptor densities among species seem to be ecologically relevant [30]. Other studies have failed to find alignment between the visual system and the behavior or ecology of species. Ultraviolet vision, for example, does not seem to have co-evolved with plumage coloration across most bird families [23, 79], but see [29]. Proper parameterization of visual models could be of paramount importance when investigating spectral tuning and the evolution of visual systems in birds. This also applies to studies of sexual dichomatism, which have been used to investigate, among other topics, the influence of sexual selection on specia- tion [54, 56]. In this study, we systematically compared the effects of changing single parameters in visual models on mean dichromatism scores, and ranks of dichromatism scores, in 70 species of Gal- liformes. We found high correlations between the mean dichromatism scores regardless of the PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 12 / 22 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models Table 4. Summary of the changes in dichromatism score of the species most affected by changes in the sensory exprience, based on the largest difference in rank. Parameter Eye type Conditions Condition 1 (< 1 JND) Condition 2 (< 1 JND) > 1 JND change Max change (JNDs) Max change (%) Light environment UVS Ideal vs D65 2 4 2 1.27 NA VS Ideal vs D65 10 4 4 1.22 NA Photoreceptor λmax UVS All Max vs All Min 2 0 2 1.12 NA VS All Max vs All Min 13 8 5 1.43 NA Oil droplet cut-off value UVS R Min vs All Max 9 7 2 1.04 NA VS R Max vs All Min 4 6 2 1.02 NA Ocular media T50 values UVS T314 vs T344 4 5 1 1.08 NA VS T335 vs T395 9 14 5 1.33 NA Photoreceptor densities UVS A. minutus vs P. paciﬁcus 8 1 12 3.62 73.72 VS A. minutus vs P. paciﬁcus 14 7 7 1.36 NA Model systems - Average UVS vs Average VS 2 10 12 2.14 120.30 - P. cristatus vs S. vulgaris 8 2 14 2.40 136.36 tism score of the species most affected by changes in the sensory exprience, based on the largest Table 4. Summary of the changes in dichromatism score of the species most affected by changes in the difference in rank. Values reported describe the number of patches (out of 15) without any discernable dichromatism (just-noticeable-differences < 1) under the ﬁrst set of conditions (Condition 1: < 1 JND) and under the second set of conditions (Condition 2: < 1 JND), the number of patches that changed by more than 1 JND when comparing the ﬁrst and second set of conditions (> 1 JND change), the maximum dichromatism value change for a single patch (Maximum change in JND) and its percentage change (Maximum change in percentage); NA values indicate that the score under one of the conditions is < 1 JNDs. See Methods and Results for more details. Light environment–Some of the earliest research aimed at assessing the importance of visual model parameters demonstrated that differences in light environments made Trinida- dian guppies (Poecilia reticulate Peters) more conspicuous in the presence of conspecifics than in the presence of heterospecific predators [80]. The importance of this parameter was further demonstrated in behavioral trials which showed that the absence of UV wavelengths reduced the foraging efficiency of three-spined sticklebacks (Gasterosteus aculeatus Linnaeus; [43]). In contrast, the light environment has almost no effect on the discriminability of vole urine against the vegetation background (~4% JNDs) suggesting little influence on the behavior of foraging raptors [22]. The results from these studies, and others (e.g., [41, 42, 81]), indicate 13 / 22 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models urnal.pone.0169810 January 11, 2017 14 / 22 PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 14 / 22 Parameterization of Avian Visual Models Fig 5. Range of sexual dichromatism scores of three galliform species obtained through exploration of the multidimensional parameter space. Distribution of mean dichromatism scores (left panels) and number of patches (out of 15) with scores greater than 1 JND (right panels), for three species, generated through permutation of all parameter conditions explored in this study (see Multidimensional parameter space section in Methods). Lines for average UVS eye type, average VS eye type, and Pavo cristatus indicate values calculated under an ideal illuminant. Fig 5. Range of sexual dichromatism scores of three galliform species obtained through exploration of the multidimensional parameter space. Distribution of mean dichromatism scores (left panels) and number of patches (out of 15) with scores greater than 1 JND (right panels), for three species, generated through permutation of all parameter conditions explored in this study (see Multidimensional parameter space section in Methods). Lines for average UVS eye type, average VS eye type, and Pavo cristatus indicate values calculated under an ideal illuminant. doi:10.1371/journal.pone.0169810.g005 that the importance of the light environment in visual models is context dependent. In our study, differences in light environment had relatively small but non-negligible effects on the mean dichromatism scores and ranks in Galliformes. The JNDs of some of the patches changed by more than 25%, differences considerably larger than those modelled by Lind et al. [22]. Our largest observed differences included the daylight 65 (D65) illuminant for both UVS and VS eye types (Table 4). This commonly-modelled light environment is rich in blue (but not UV) wave- length and is likely to influence colors rich in red and UV wavelength such as those produced by some carotenoids [82]. Also of great importance is the consideration that color constancy mechanisms, approximated by the von Kries correction in the receptor noise model, limit the influence of the light environment on the perceived contrast between color patches. Behavioural tests [83] and modelling experiments [84] in vertebrates have shown that chromatic adaptation can work over large changes in ambient light environments. Photoreceptor sensitivities–In a test of parameterization effects on models in foraging rap- tors, Lind et al. [22] reported chromatic differences of ~16% ± 12% JNDs when comparing a range of photoreceptor sensitivities. These results were obtained by changing the SWS1 and SWS2 photoreceptor by 10 nm towards short-wavelength sensitivity and the LWS by 10 nm towards long-wavelength sensitivity. Our general results support these findings and also dem- onstrate that differences greater than ~23% are possible. These values were obtained by com- paring all minimum-shifted and all maximum-shifted photoreceptor sensitivities (Table C in S3 File). However, changes in the sensitivity of single photoreceptors within eye type, even to the extreme known values across birds, had very little influence on dichromatism scores (Table I in S3 File). These results also support work presented by Lind and Kelber [44], which found little influence of photoreceptor sensitivity in modelling chromatic differences between four colors (peak wavelength at 350, 450, 500, and 650 nm) against a green background. Over- all, the parameterization of photoreceptor sensitivities should only have consequential influ- ences on chromatic contrast calculations when all sensitivities are wrongfully shifted in the same direction (all towards short- or long-wavelengths) or when SWS and LWS photorecep- tors are shifted in opposite directions. In contrast, changes in single photoreceptor sensitivities generally have limited effects on calculated JND scores, but some consequential effects on dichromatism ranks. Photoreceptor sensitivities–In a test of parameterization effects on models in foraging rap- tors, Lind et al. [22] reported chromatic differences of ~16% ± 12% JNDs when comparing a range of photoreceptor sensitivities. These results were obtained by changing the SWS1 and SWS2 photoreceptor by 10 nm towards short-wavelength sensitivity and the LWS by 10 nm towards long-wavelength sensitivity. Our general results support these findings and also dem- onstrate that differences greater than ~23% are possible. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Parameterization of Avian Visual Models revealed that dietary carotenoid content can influence the transmission properties of oil drop- lets in double cones, indicating condition-based within-species variation in visual properties and the potential of diet to influence color vision [86]. To date, the influence of variation in oil droplet characteristics on color discrimination have only been modelled, never behaviorally tested. Ocular media–The only other study to have explicitly modelled the influence of ocular media on chromatic contrasts found this parameter rather inconsequential [44]. Our overall results (Table 2) agree with these findings but highlighted that this parameter is perhaps most important when modelling VS eye types, not UVS eye types (Table 4). Differences in this parameter will only influence the perception of UV-rich colors which are common in fruits and feathers that contain carotenoids [82, 87], vole-urine used by foraging raptors to assess prey density [22, 88], and some structural colors (e.g.,[89–91]). Particular consideration to this parameter should be made when modelling color discrimination of these UV rich colors. Photoreceptor densities–The photoreceptor density was the single most important parame- ter in our models both in terms of changes in mean dichromatism ranks and chromatic contrast of individual patches (Tables 2 and 4). This occurs because the Weber fraction is proportional to the inverse of the square root of the photoreceptor ratio (see formula in supplemental mate- rial in [12, 75]), and has a linear relationship with JNDs. For example, a dichromatic score of 6 JNDs for a Weber fraction of 0.05 will be reduced to 3 JNDs with a Weber faction of 0.10, and further reduced to 1.5 JNDs with a Weber fraction of 0.20. It is perhaps not surprising then, that photoreceptor density has such a large influence on chromatic contrast thresholds. Individual patches changed by as much as 10.5 JNDs (~50%), values in the range of to those presented by Lind and Kelber [44], which demonstrates the importance of this visual system trait. Differences in photoreceptor densities are likely to have large consequences on among-species ability to dis- criminate between similar colors and, as for variation in oil droplet absorbance profiles, within- species variation may be of consequence as well. For example, differences in densities among house sparrows (Passer domesticus Linnaeus) generated chromatic contrast differences of ~16% (> 3 JNDs) when evaluating the perception of the white wing bars against the brown wing back- ground coloration [45]. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 These values were obtained by com- paring all minimum-shifted and all maximum-shifted photoreceptor sensitivities (Table C in S3 File). However, changes in the sensitivity of single photoreceptors within eye type, even to the extreme known values across birds, had very little influence on dichromatism scores (Table I in S3 File). These results also support work presented by Lind and Kelber [44], which found little influence of photoreceptor sensitivity in modelling chromatic differences between four colors (peak wavelength at 350, 450, 500, and 650 nm) against a green background. Over- all, the parameterization of photoreceptor sensitivities should only have consequential influ- ences on chromatic contrast calculations when all sensitivities are wrongfully shifted in the same direction (all towards short- or long-wavelengths) or when SWS and LWS photorecep- tors are shifted in opposite directions. In contrast, changes in single photoreceptor sensitivities generally have limited effects on calculated JND scores, but some consequential effects on dichromatism ranks. Oil droplet cut-off value–Variation in the transmission cut-off values of oil droplets had sim- ilar but potentially slightly larger influence on dichromatism scores than variation in photore- ceptor sensitivities. Our results demonstrate that differences greater than 41% (> 7 JNDs in this case) in single patch chromatic contrasts are possible. Lind and Kelber [44] also demonstrated the importance of this parameter. In a behavioural experiment comparing the measured and predicted visual sensitivities in domestic pigeons (Columba livia), the match between visual models and behavioral results in tests of hue discriminability improved tremendously by shift- ing the absorbance curves of oil droplets by 10 nm in their models. Oil droplet properties of the avian eye should perhaps be given greater attention. Indeed, considerable variation has been found within species, both among individuals and between the sexes [85]. Modelling of within- species differences suggest chromatic contrast differences as large as ~30% JNDs in some parts of the visual spectrum [48], sufficient to influence the perspective of the receivers, and poten- tially affecting foraging and mate choice behaviors. Furthermore, recent experiments have PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 15 / 22 In certain instances, two or more parameters may cancel each other out (e.g., a short-wavelength shifted SWS1 value matched with a long-wavelength shifted ocular media value), but other combinations of incorrect parameters could dramatically alter calculated val- ues. For example, a short-wavelength-shifted SWS1 photoreceptor sensitivity value matched with a photoreceptor density ratio that favours discriminability in the short wavelengths could mistakenly modify a UV-insensitive visual system to one that can detect small color differences in the UV range. In extreme cases, as demonstrated by the permutation scores calculated for T. tibetanus, improper parameterization could lead to differences between calculated and actual contrasts of more than 650%. As for errors in single parameters, errors in multiparameter space are more likely to affect the scores of species with low dichromatism scores, and models that generate values near threshold should be interpreted with caution. Minimizing uncer- tainty in the most influential parameters, namely photoreceptor densities and peak sensitivity of the SWS1 photoreceptor, should increase confidence in the calculated dichromatism scores. Caution is also needed when making inferences regarding large contrast values in units of just-noticeable-differences. Indeed, the primary function of the receptor noise model is to com- putationally determine discrimination thresholds, based on a psychophysical model [12]. As such, the meaning of supra-threshold distances is generally unknown. Is a color patch differing by 5 JNDs as discriminable from a reference as a patch differing by 7 JNDs? Following Fechner’s assumption of additivity, large colorspace should be possible to model (discussed in [95, 96] among others). Furthermore, the only study investigating a behavioral response (in this case the visual attention reflex) in relation to perceptually large color distances found that just-notice- able-differences, as calculated by the receptor noise model, are effective in describing colors sep- arated by large distances in visual space [97]. Nevertheless, because the evidence is sparse, very large differences in units of JND should be interpreted with prudence. It is also important to consider that animals do not perceive individual patches in isolation [24] and that the contrasts among several color patches may be evaluated differently than the sum of its parts, concepts that are currently being investigated [98]. Whether or not the mean (or sum) of dichromatism scores are meaningful beyond a discrimination threshold will need further study. Even if these plumage patches likely differ more in the achromatic com- ponent of the signal, these chromatic differences may still have implications for mate choice and agonistic interactions. Because the characterization of complete visual systems requires spe- cialized equipment and skills, and the sacrifice of animals, our knowledge of photoreceptor den- sities come from relatively few studies (see Hart [30] for the majority of species characterized to date). Future research on the physical properties of avian retinas should obtain as much infor- mation as possible, including counts of different photoreceptor types. Just as importantly, the assumptions that 1) relative receptor noise is truly proportional to receptor abundance and that 2) receptor noise is independent of receptor type need to be rigorously tested. These data have the potential to make large contributions to our understanding of the visual ecology of birds. Model systems–Within eye types, there was relatively small difference in mean dichroma- tism scores among the model visual systems. However, our results suggest that one of the most influential parameters of visual models is whether a species possesses a UVS or VS eye type (Tables 2–4, Table M in S3 File). This is of particular importance since these are the two most commonly used sets of parameters in avian visual modelling. Because there was a strong belief in phylogenetic inertia in eye type (e.g., [25, 26]), studies have usually modelled a single eye type (exceptions include: [92–94]). However, as demonstrated by Renoult et al.[27], using the wrong eye type can entirely alter the conclusions of a study. Fortunately, determination of a species’ eye type does not require microspectrophotometry, as opsin gene sequences can be used to estimate sensitivity [33]. Even though not all SWS1 opsin gene sequence variations have been compared to measured photopigment sensitivities, the peak absorbance of short- PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 16 / 22 Parameterization of Avian Visual Models wavelength photoreceptor can usually be estimated [28, 33]. This method is relatively rapid, inexpensive, and could easily be implemented in any molecular laboratory [33]. Alternatively, when this is not possible, both eye types could be modelled when the eye type is not known. Without a doubt, sensory perceptions are produced by the combined physical properties of sensory systems. By calculating the range of mean dichromatism scores for all parameter com- binations, we demonstrated that misinformed parameterization could lead to very different conclusions. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Overall, our results suggest that a single parameter can have large influences on avian dichro- matism scores calculated by combining the chromatic contrast of several plumage patches, and the rank position of a species and/or their color patches in relation to one another. Further- more, we demonstrated that contrasts calculated by varying multiple parameters can generate extreme discrepancies between estimated dichromatism scores. To improve the reliability of avian visual models, information about photoreceptor densities and the sensitivity of the SWS1 photoreceptor should be investigated when possible. Because sequencing the SWS1 gene is cost- effective, we recommend that researchers modelling avian visual systems determine, at least, whether their species of interest possess a UVS or VS eye type. Alternatively, results should pres- ent analyses from both eye types to confirm that the findings are robust. When little visual sys- tem information is available for a species of interest, in birds and other systems, reporting the results of sensitivity analyses would help strengthen the inferences presented. 17 / 22 Parameterization of Avian Visual Models Supporting Information S1 File. List of Abbreviations. (DOCX) S2 File. Formulae. (DOCX) S3 File. Supporting information. (DOCX) S4 File. Average spectral curves from 3 males, and 3 females, from 70 species of Galliform, 15 plumage patches per species. (CSV) S5 File. Species by plumage patch matrix indicating which patches are bare (patches not used in analyses). (CSV) S4 File. Average spectral curves from 3 males, and 3 females, from 70 species of Galliform, 15 plumage patches per species. (CSV) S5 File. Species by plumage patch matrix indicating which patches are bare (patches not used in analyses). (CSV) S5 File. Species by plumage patch matrix indicating which patches are bare (patches not used in analyses). (CSV) Acknowledgments We thank the University of Michigan Museum of Zoology, the Field Museum of Natural His- tory, the American Museum of Natural History, and the Royal Ontario Museum for providing access to museum specimens. We thank the University of Michigan Museum of Zoology, the Field Museum of Natural His- tory, the American Museum of Natural History, and the Royal Ontario Museum for providing access to museum specimens. Funding acquisition: PPB KJ SMD. Investigation: PPB KJ. Methodology: PPB. Methodology: PPB. Supervision: SMD. Supervision: SMD. Visualization: PPB. Writing – original draft: PPB. Writing – review & editing: PPB KJ SMD. Author Contributions Conceptualization: PPB SMD. PLOS ONE \| DOI:10.1371/journal.pone.0169810 January 11, 2017 Conceptualization: PPB SMD. Formal analysis: PPB. Formal analysis: PPB. Funding acquisition: PPB KJ SMD. References 1. Limeri LB, Morehouse NI. 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https://openalex.org/W4388659464	OpenAlex	Open Science	CC-By	2,023	Community-Based Village Management Independence In Efforts to Create Sustainable Tourism (Panglipuran Tourism Village Phenomena)	Agung Sulistyo	English	Spoken	6,711	11,071	Community-Based Village Management Independence In Efforts to Create Sustainable Tourism (Panglipuran Tourism Village Phenomena) Agung Sulistyo, Indah Fatmawati and Nuryakin Doctoral Program of Management, Muhammadiyah University of Yogyakarta, Indonesia Agung Sulistyo, Indah Fatmawati and Nuryakin Doctoral Program of Management, Muhammadiyah University of Yogyakarta, Indonesia Abstract. Tourist behavior change in choosing tourist attractions have made tourism villages the prima donna. The presence of a tourist village is interpreted as an integration between interest and social activity with the leading role played by the community. Through these activities, the part of tourism in community empowerment is something that needs attention. This study aims to find the meaning of Independence in managing tourist villages carried out by the community. This study uses a qualitative design through a phenomenon. This study tries to explore the community's experience as a tourist village manager and the impacts that arise. The data collection process was carried out through semi-structured interviews with five informants. This research showed the meaning of community independence and its impact on managing tourist villages, including community self- confidence, community commitment, community economic empowerment and knowledge transfer. Thus, good management will provide positive results for the village's and its people's development. Several studies strengthen the argument for the role of the community as managers of regional potential. However, only some studies have raised the issue of community independence in managing tourist villages. Besides that, the conclusions of this study can also provide the guidance needed to develop regional potential, especially in tourism villages. , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (https://creativecommons.org/licenses/by/4.0/). 1 Introduction Through community empowerment, tourism is essential in developing regional potential [1–3]. More attention must be given to all parties in tourism development and its managing operator [4]. One of the developments in the tourism sector is marked by the presence of tourist villages as an alternative tourist destination. According to data released by the Ministry of Tourism and Creative Economy of the Republic of Indonesia, tourist behaviour has changed. This situation can be seen from the choice of tourist destinations which have shifted from mass tourism to alternative tourism, such as tourist villages [5]. This data is strengthened by a release issued by the Ministry of Villages of the Republic of Indonesia regarding the number of potential villages reaching 7,725 [6]. It is interesting to see how * Corresponding author: agung.sulistyo.psc20@mail.umy.ac.id * Corresponding author: agung.sulistyo.psc20@mail.umy.ac.id © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (https://creativecommons.org/licenses/by/4.0/). , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 the growth of tourist villages in Indonesia has experienced significant developments in terms of numbers. It is okay if great hopes are pinned on the management of tourist villages [7,8]. the growth of tourist villages in Indonesia has experienced significant developments in terms of numbers. It is okay if great hopes are pinned on the management of tourist villages [7,8]. Tourism villages are interpreted as acculturation between attractions, social life and the community as the leading managers [9,10]. In its development, the management of tourism villages is divided into several levels: pilot, developing, advanced, and independent [11]. Based on data released by the Ministry of Tourism and Creative Economy of the Republic of Indonesia in 2021, there are around 1,836 tourist villages. This number is divided into several tourism village clusters, including the pilot tourism village category (709 tourism villages), the developing tourism village category (967 tourism villages), the advanced tourism village category (158 tourism villages) and the independent tourism village category (2 tourism villages). This number will increase significantly in 2022 to 3,419 tourist villages. The Ministry of Tourism and Creative Economy received severe attention through the Indonesian Tourism Village Award (ADWI) [6]. g g ( ) Subsequent developments emphasize the vital role of the community in maximizing the potential of a tourist village [12,13]. Several studies strengthen the argument for the role of the community as managers of regional potential [14,15] so that they can maximize and empower the community's economy [12,16]. However, only some studies have raised the issue of community independence in managing tourist villages. This study explores the extent of community participation in developing the Panglipuran tourism village. Panglipuran tourism village has an attraction that can become a magnet for both local and foreign tourists in the province of Bali [17,18]. The findings in this study can provide policy and impact information and implications for the public. Besides that, the conclusions of this study can also provide the guidance needed to develop regional potential, especially in tourism villages. 2.3 Independence Independence is defined as the ability to rely on one's cabilities in a manner individuals to meet their needs [44]. Other literature explains that Independence is defined as a person's ability to meet basic needs such as security, education, health and the need to interact and socialize [45–47]. Independence is essential developing quality of society in the environment. Furthermore, the community needs to position itself and empower the region's potential [12]. As the prime mover, society's quality will face various problems. A self-concept is needed to be a driving force in tourism development [48]. Research on the Independence of the management of tourist villages has yet to be widely carried out, so this study is expected to fill the gaps in research on the impact of the development of tourist villages on community independence. 2.1 Community-Based Tourism Community-based tourism is essential to regional empowerment and development [19,20]. The emergence of these activities as a management response ignores environmental factors [21,22] and environmental authenticity [23,24]. Community empowerment as an instrument for managing tourism is not only in the area of village empowerment but has a broader scope [14]. Several additional areas of activity that can empower the community include the arts [25], culinary [26], to local cultural acculturation [27]. This study uses the terminology of tourism development through community participation as the prime mover. Community empowerment in tourism management will present various attractive offers to create prosperity and strengthen the economy. Thus, one of the sustainable tourism instruments will be made through the presence of the community as a drive. The role of the community in tourism management is increasingly widespread and recognized by many countries [20,28]. This scheme can improve the quality and welfare of the community's economy. Several studies have strengthened the role of the community in managing and developing the regional economy [12,16]. Community-based tourism management embodies cultural wealth and agreements produced within the community. Management is carried out using mechanisms regulated by local community groups. The deals creates harmony and indicates the presence of tourists to policymakers [29,30]. However, community-based management needs help. Some inhibiting factors are differences in point of view [27] and management capabilities that have not been maximized [31]. Even though many parties give attention, the community should not be too 2 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 dependent. Communities must be independent and be determined to develop amidst existing limitations [20]. dependent. Communities must be independent and be determined to develop amidst existing limitations [20]. 2.2 Tourism Village The tourism sector is experiencing developments in line with technological changes [32,33], tourist behaviour [34] to the attractions offered [35]. Management and marketing activities have also changed and can be seen from the opportunities and threats that come [36]. Various tourist attractions have emerged, ranging from cultural tourism, nature tourism, and agricultural tourism [35] to tourist villages that have distinctive characteristics and engaging social activities [11,37]. These attractions are valuable capital in community empowerment and efforts to increase the regional economy. On the other hand, sustainability in tourism must consider the economic and social environment and maintain its attractiveness. [38]. Tourism villages are increasingly showing their existence as new potentials in tourism development. Tourists are interested in the community's ability to manage tourism villages [39,40]. The terminology of tourism villages is interpreted as the relationship between attractiveness, accommodation and other things offered in a social system that is continuously maintained [11,37]. The development of tourist villages is increasingly competitive, as seen from their various potential to attract tourist visits [41]. The community's ability to manage and design a competitive tourism village must understand and adapt to various situations [29]. Various obstacles, such as consistency, originality and human resources, can be resolved [42]. It is interesting to know how a tourist village can solve many things contrary to the authenticity of the environment and create an understanding in the middle public [21,29,43]. 3.2 Object of research This study uses the Panglipuran tourism village as a research object. This tourist village is in Bangli Regency, Bali Province, Indonesia. Researchers chose this tourist village because of several achievements and unique characteristics. Various awards have been won by the Penglipuran tourism village, including the Asian Homestay Standard Tourism Village and the 2019 Green Destination Sustainable [54], as well as a non-tourism award as a Climate Village. This tourist village is also unique in the design of existing residential buildings and the strong community commitment to its management. 2.4 Community Empowerment People must have the skills and abilities to create prosperity and improve their quality of life [49]. Independence is interpreted as an activity to increase the confidence of both individuals and groups towards a better situation [50]. Through community empowerment, each individual and group will try to develop various things, including self-confidence [51]. Assignments involving the community can become a strong foundation for developing regional potential. Empowerment provides an opportunity for the community to design various things to meet their needs and make decisions to take action based on their initiatives. Thus, creating community-based sustainable tourism is something that can be realized. 3 3 https://doi.org/10.1051/e3sconf/202344401013 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 3.1 Research Design This study uses a qualitative research design through a phenomenon. The interpretive philosophy was chosen to explore the various knowledge and experiences of the informants [52]. This study will analyze how practices and behavior occur. This stage will ensure that the inspection will be carried out thoroughly and in detail to get results that describe various factors [53]. This research seeks to see the community's Independence in managing tourism villages, developing and maintaining each process towards continuity. 3.4 Data analysis This research explores the informants' various experiences managing the Panglipuran tourism village and the resulting impact. The interview process in this study was recorded using a tape recorder and analyzed using the Phenomenological Interpretative Analysis method. This analysis is used to inspect the experience of life someone disclosed in their terms of use [57]. This analysis becomes a guide through to phenomena and their experiences _ experience in detail [58]. 3.3 Data Collection This study used a semi-structured interview technique with submitted questions to the public as informants. Teach This is to dig a variety of information and their experience. Various questions were prepared and asked to guide the interview process. The questions in the interview process were based on field observations and were supported by a literature review to obtain targeted research results [33,55]. Eighteen main interview questions were asked during the ongoing interview process. This activity created an atmosphere of dialogue between researchers and informants to find answers and explore various other essential aspects. Table 1. Research Informants No Code Gender Role 1 2 3 4 5 I1 I2 I3 I4 I5 Man Man Man Man Woman Customary Institution Service Institution Tourism Village Manager Security / Pecalang Public Quality information must be supported by the determination of good informants [56]. Researchers have determined informants who are considered appropriate and represent the community. This study used five informants as research participants. The informants are people who have experience and are considered to be the driving force for creating community-based tourism village management. Informants in this study are parties who have direct involvement in the process of forming and managing villages tour. Researchers acquainted with the informants in the early stages of selecting data search. To strengthen the search for data, researchers settled for three days in the Panglipuran tourist village. This activity was carried out to establish communication and make further observations. The informants consisted of leaders of traditional institutions (I1), heads of service agencies (I2), managers of tourist villages (I3), security or pedalling (I4) and local communities (I5). 4 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 The informants are representatives of the community who are considered suitable as a source of data collection. 4.1.1 Pride The existence of tourist villages is currently experiencing quite good development. This condition shows changes in the behavior of tourists who make the village a tourist destination. Tourists are attracted by various authenticity and social activities carried out by the local community. This situation directly has a positive impact on society. They are proud of their conditions and actions that can give tourists a sense of wonder. "We are proud of what we have even though the development outside is high-speed" (I1, I2) ( , ) "Tourists respect various things in this village, such as culture and life " (I3) "Now tourists like to come here; they see our activities and all about our culture " (I5) 4 Result and Discussion The informants shared their experiences about their arguments against the management of the Panglipuran tourism village. This research focuses on self-management carried out by the community. The independence model found in the direction of Panglipuran Tourism Village includes (1) community confidence, (2) community commitment, (3) community empowerment and (4) knowledge transfer. Visually, community independence is presented in the following table: Table 2. The Impact of Community Independence on the Management of Tourism Villages No Community Independence Impact Table 2. The Impact of Community Independence on the Management of Tourism Villages Table 2. The Impact of Community Independence on the Management of Tourism Villages No Community Independence Impact 1 Community Confidence Pride Social status 2 Community Commitment Cooperation Problem-solving Area Development 3 Community Economic Empowerment Employment Community Economic Welfare 4 Knowledge transfer Comparative study Cultural Preservation 4.2.1 Cooperation As one of the essential instruments for a tourist village, an understanding of the community needs to be maintained. Various arguments and desires to achieve a common goal. The Panglipuran community is committed to advancing their village through tourism activities without eliminating the existing culture. These conditions match the representation of their village name Panglipuran, which means remembering their ancestral land. As a manifestation of this, they work together to achieve goals that align with the existing culture. "Village people are full of tolerance and cooperation; this needs to be maintained" (I2) "Village people are full of tolerance and cooperation; this needs to be maintained" (I2) g p p p "If there is still an individual ego out there, then this is not allowed here (I3, I4) "living in a village like this, helping each other neighbours or cooperation in other matters" (I5) 4.2.2 Problem-solving Like most other communities, village communities cannot be separated from social problems because each individual has different desires and opinions. These situations sometimes create friction that needs to be resolved. However, Panglipuran tourism village has a way to overcome this problem. They have a particular schedule and place to discuss solving problems. Thus, the seeds of differences of opinion can be resolved quickly. solving problems. Thus, the seeds of differences of opinion can be resolved quickly. "It is normal for people to have differences or problems, but we will solve them as much as possible according to custom" (I1-I2) "It is normal for people to have differences or problems, but we will solve them as much as possible according to custom" (I1-I2) "Village people are like that, solving problems g p p , g p y g g Periodically, the people here gather to discuss various matters" (I3) Periodically, the people here gather to discuss various matters" (I3) "Problems will be resolved, we have a regular schedule for gathering, and it is usually done at the meeting hall (I4, I5) "Problems will be resolved, we have a regular schedule for gathering, and it is usually d t th ti h ll (I4 I5) "Problems will be resolved, we have a regular schedule for gathering, and it is usually done at the meeting hall (I4, I5) 4.1.2 Social status The existing community in a tourist village becomes the driving force of tourism go continuation. How could they not? The different things they are doing are now bearing 5 5 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 fruit. They are no longer ashamed of their status as village people but are now considered village people. The various things they have tried and maintained are currently having a positive impact. Their daily activities are things tourists want to know. Panglipuran tourism village which is increasingly famous directly raises the dignity of the village community 4.3.1 Employment This place opens opportunities for local people to become part of village management (I5) "For the community, the presence of a tourist village plays a role in the continuity of h i li h i i i i l d h " (I2) "Indeed, not all of the people are absorbed, some want to migrate, and some want to survive because there are jobs they can do here" (I3) "We are security officers here, guarding the activities and smooth running of Penglipuran village" (I4) "Being a foreigner also exists; surviving here as a manager is also a lot. Everyone has the right to determine whether to live here or elsewhere. This place opens opportunities for local people to become part of village management (I5) 4.2.3 Area Development Tourism broadly impacts various sectors, including empowerment and regional development. The presence of a tourist village is interpreted as the implementation of sustainable tourism development. Tourist behavior that changes from mass tourism to special interest tourism is one of the drivers of the development of tourism villages. This opportunity certainly needs to be seized by various parties opportunity certainly needs to be seized by various parties. "Tourism is one of our ways to develop the region" (I1, I2) "This village is growing, and the presence of tourists is one of the reinforcements (I3) i hi i l b d i ill d l ( ) "Was in this area is only forest, but turned into villages and settlements (I4) 6 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 "The village community will accept the contribution made by tourists. We are assisted in renovating and repairing other facilities in this village" (I5) 4.3.2 Community Economic Welfare The increasingly strong tourism sector stimulates the presence of tourism-supporting instruments. Transportation, lodging, and souvenirs are several things in supporting sustainable tourism. The instrument can be carried out and provided by the local community. Thus, the economic potential will also be felt by the community. Showing what they have is a pride for themselves. The role of official and customary institutions is crucial in making policies and regulating various matters. Activities usually carried out by the village community now have another impact on their lives. g y p "The community can provide lodging for tourists who want to feel life in the village for a long time" (I1) "Some people sell typical products, souvenirs or other things. This situation directly impacted the economic life community (I2) "In this village, there is also a kind of traditional market, but many people also sell various products in their homes" (I3) "I sell several products such as drinks, food and some souvenirs. The results of the sale can add to family finances" (I5) "The community can provide lodging for tourists who want to feel life in the village for a long time" (I1) g ( ) "Some people sell typical products, souvenirs or other things. This situation directly impacted the economic life community (I2) "In this village, there is also a kind of traditional market, but many people also sell "Some people sell typical products, souvenirs or other things. This situation directly impacted the economic life community (I2) "In this village, there is also a kind of traditional market, but many people also sell various products in their homes" (I3) "I sell several products such as drinks, food and some souvenirs. The results of the sale can add to family finances" (I5) 4.3.1 Employment Another impact of tourism development is the ability to absorb the community as managers. This condition indirectly becomes a solution for the government in creating jobs. The village community is the driving force as well as the manager of the attractiveness they have. People no longer need to leave their area to get a job. They can carry out activities while fulfilling their needs in their territory. Their pride in the village will be even more tremendous. "This tourist village can absorb a productive workforce; the local village community runs various positions" (I1) "For the community, the presence of a tourist village plays a role in the continuity of their lives; they can carry out various activities related to the economy" (I2) "Indeed, not all of the people are absorbed, some want to migrate, and some want to survive because there are jobs they can do here" (I3) "We are security officers here, guarding the activities and smooth running of Penglipuran village" (I4) "Being a foreigner also exists; surviving here as a manager is also a lot. Everyone has the right to determine whether to live here or elsewhere. This place opens opportunities for local people to become part of village management (I5) "This tourist village can absorb a productive workforce; the local village community runs various positions" (I1) "This tourist village can absorb a productive workforce; the local village community runs various positions" (I1) "For the community, the presence of a tourist village plays a role in the continuity of their lives; they can carry out various activities related to the economy" (I2) "Indeed, not all of the people are absorbed, some want to migrate, and some want to survive because there are jobs they can do here" (I3) "We are security officers here, guarding the activities and smooth running of Penglipuran village" (I4) "Being a foreigner also exists; surviving here as a manager is also a lot. Everyone has the right to determine whether to live here or elsewhere. 4.4.1 Comparative Study The ability to manage a good tourist village will have a positive impact not only on the local community but also on the broader community. Panglipuran tourist village, which has gone worldwide, is one of the capitals for creating continued success. This condition can be seen from the number of visits by tourists. Visits are not only made by individual tourists but also in groups, Good domestic and foreign. Along the way, many visits were made by 4.4.2 Cultural Preservation Tourists get many things when they visit tourist villages. Some of them can see the community's social activities and enjoy the arts to see the still upheld culture. Tourism can develop but not destroy the culture that is already rooted. The Panglipuran tourism village community adheres to this principle. Following the meaning of the name Panglipuran, namely remembering the ancestors, the local community always respects the culture that has existed for a long time. Various things and traditions that exist are still carried out according to belief. The community's ability to maintain cultural values and practices is a unique tourist attraction. Tourists see various things related to Panglipuran village culture and differences from their area. This condition is also interpreted as a way for the Panglipuran village community to introduce their culture. "We have customary rules that must be maintained, respected and implemented. These various things are one of the ways to maintain the sustainability and harmony of society" (I1, I2) "We have customary rules that must be maintained, respected and implemented. These various things are one of the ways to maintain the sustainability and harmony of society" (I1, I2) "We have customary rules that must be maintained, respected and implemented. These various things are one of the ways to maintain the sustainability and harmony of society" (I1, I2) "Panglipuran tourism village has customs and culture that are upheld; tourists admire this kind of community behaviour" (I3) "Various behaviours and aspects of life have been regulated by tradition from generation to generation. If there are problems, they can be resolved with the existing provisions" (I4) "Panglipuran tourism village has customs and culture that are upheld; tourists admire this kind of community behaviour" (I3) "Various behaviours and aspects of life have been regulated by tradition from generation to generation. If there are problems, they can be resolved with the existing provisions" (I4) "We have a culture that must be maintained and preserved. Our different culture is interesting for tourists to know and learn about it" (I5) "We have a culture that must be maintained and preserved. Our different culture is interesting for tourists to know and learn about it" (I5) This research shows that the Panglipuran village community has Independence. This situation is reflected in several aspects, including community confidence, commitment, economic empowerment and knowledge transfer. 7 7 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 groups managing tourism villages in other areas. This condition further emphasizes the existence of the Panglipuran tourist village. g p g " Panglipuran tourist village is visited by many tourists, both local and foreign. They want to see the real face of Panglipuran" (I1) " Panglipuran tourist village is visited by many tourists, both local and foreign. They want to see the real face of Panglipuran" (I1) "Panglipuran is frequently visited by tourist village groups from areas outside the province of Bali. They want to see community life and village management" (I2) "Panglipuran is frequently visited by tourist village groups from areas outside the province of Bali. They want to see community life and village management" (I2) "I have had discussions with groups from Jakarta, Kalimantan and outside Indonesia, such as Japanese, American and several other countries. They come here to see the activities of the people here" (I3) a g pu a s eque t y v s ted by tou st v age g oups o a eas outs de t e province of Bali. They want to see community life and village management" (I2) "I have had discussions with groups from Jakarta, Kalimantan and outside Indonesia, such as Japanese, American and several other countries. They come here to see the activities of the people here" (I3) "I have had discussions with groups from Jakarta, Kalimantan and outside Indonesia, such as Japanese, American and several other countries. They come here to see the activities of the people here" (I3) "Tourists who come ask many things such as; life, worship, activities to other things. They want to know various things in Panglipuran village" (I5) 4.4.2 Cultural Preservation The local community firmly holds Independence and culture that go hand in hand with developing tourist villages. These things have been passed down from generation to generation as things that must be maintained. Following the meaning of Panglipuran, namely remembering their ancestors, the activities of the local community cannot be separated from the culture still being preserved. Development impact _ tourists in a manner that will directly benefit the region's development [27]. However, these developments continue the community's traditions to preserve their culture. Even though the community lives in the village, their ability to manage the area has a positive impact. Their success in maintaining tradition and culture gives confidence to the local community. In this condition, they believe that culture does not hinder tourism development [13]. The two are complementary parts of each other. Directly, the village community is no longer assessed as being left behind. They live during changing times but still maintain cultural values. Tourism turns their lives into a positive value full of meaning and pride. Few village people have changed their social status with tourism development, primarily through tourism villages. 8 8 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 Tourism has a significant impact on regional development. The emergence of various new tourist attractions marks this condition. This development can be interpreted as an effort to develop its potential. The multiple efforts made are implementing agreements in the community. As the primary mover, the presence of the community as a manager is something that needs attention [59]. The cooperation carried out by the community aims to create harmony and comfort in life. However, living in society is challenging. This condition also applies to the management of the Panglipuran tourism village. Leaders at the level of traditional and official institutions pay attention to this. They make use of rules that have been used for generations. The resolution of conflicts that exist in the community is driven by deliberation [56]. To reduce conflicts or differences in opinion, they have regular meetings to hear various things the village community wants to convey. This policy is carried out to maintain harmony in society. Sustainable tourism is tourism that can have a broad impact. The development of tourism provides many opportunities that the community can achieve [14]. 4.4.2 Cultural Preservation Tourism offers a change to be absorbed into the world of work. Tourism opens opportunities for the community to become a driving force for tourism. A similar situation also occurred in Panglipuran Tourism Village. The community has jobs from the growing development of tourism in the area. Some of the activities that the community can carry out include: being a manager, being a guide or providing various things tourists need. Tourists not only come to enjoy the attraction but also need multiple items such as lodging, transportation or souvenir products [60–62]. The local community can provide all the needs of these tourists. The village community ultimately enjoys improvements and improvements in the economy. This condition is one of the proofs that the development of regional potentials will ultimately have an impact on people's welfare. y p p p The various positive impacts of potential regional development are undeniable evidence. The success of a region in developing a variety of things can attract the attention of all parties [12]. These results become a stimulus for various parties to follow the process carried out. As a tourist village with achievements and uniqueness, Panglipuran tourism village can become a magnet for tourists to visit. They do not just come to enjoy the attraction but learn many things like The characteristics and uniqueness of buildings, community activities, and culture. This tourist village also proves that the development of the era remains the same traditions and culture they maintain. They continue to carry out activities by placing culture as their characteristic. The development of tourism and regional development continues their habit of carrying out various traditional and cultural rituals that they usually do. It is interesting to see how in the end, culture becomes one of the attractions of tourism that is not eroded by time. Community empowerment is the primary driver of regional development and aims to improve the quality of society [56]. The community will benefit from the products taking place in their area. In the end, the collaboration of various elements of society creates understanding and agreement in the community. The management of tourist villages needs to consider the benefits that the community will obtain. As the prime mover, the community must think critically but constructively about existing developments. The required village development pattern is good and gives positive results [56,63]. 4.4.2 Cultural Preservation Some of the models found in this study can be used, including community confidence, community commitment, community economic empowerment, and knowledge transfer. This study can provide suggestions for the development of literature or the management of tourist villages. This research has limitations in exploring the experiences of the Panglipuran tourism village community. Therefore, the researcher proposes the need to conduct qualitative research based on a phenomenological approach to look at the characteristics and culture of the community. This research is focused on tourist villages and investigating community 9 9 , 010 (2023) E3S Web of Conferences IConARD 2023 13 444 https://doi.org/10.1051/e3sconf/202344401013 life or other social groups. This research can be used as a starting point or literature in community empowerment efforts. life or other social groups. This research can be used as a starting point or literature in community empowerment efforts. 5 Conclusion and Recommendation The collaboration of various elements of society creates understanding and agreement in the community. The management of tourist villages needs to consider the benefits that the community will obtain. As the prime mover, the community must think critically but constructively about existing developments. 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W4238652451.txt_1	Open-Science-Pile	Open Science	Various open science	2,020	Enclomiphene Citrate	None	English	Spoken	138	291	Qeios · Definition, February 7, 2020 Ope n Pe e r Re v ie w on Qe ios Enclomiphene Citrate National Cancer Institute Source National Cancer Institute. Enclomiphene Citrate. NCI T hesaurus. Code C61745. T he orally bioavailable citrate salt of enclomiphene, the trans-isomer of the nonsteroidal triphenylethylene compound clomiphene, with tissue-selective estrogenic and antiestrogenic activities. As a selective estrogen receptor modulator (SERM), enclomiphene binds to hypothalamic estrogen receptors, blocking the negative feedback of endogenous estrogens and stimulating the release of gonadotropin-releasing hormone (GnRH) from the hypothalamus; released GnRH subsequently stimulates the release of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) from the anterior pituitary, resulting in ovulation. In addition, this agent may bind to estrogen receptors on breast cancer cells, resulting in the inhibition of estrogen-stimulated proliferation in susceptible cell populations. Qeios ID: 07Z56Z · https://doi.org/10.32388/07Z56Z 1/1.	19,408
https://openalex.org/W2898994401	OpenAlex	Open Science	CC-By	2,018	Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal	Ana Paula Wyse	English	Spoken	12,826	23,341	RESEARCH ARTICLE Ana Paula WyseID1, Antonio Jose´ Boness dos Santos1, Juarez dos Santos Azevedo2, Josenildo Silva de Lima3, Jairo Rocha de Faria1 1 Department of Scientific Computing, Informatic Center, Universidade Federal da Paraı´ba, João Pessoa, Paraı´ba, Brazil, 2 Exact and Technology Sciences Center, Universidade Federal do Recoˆncavo of Bahia, Cruz das Almas, Bahia, Brazil, 3 Graduate Program in Mathematical and Computational Modeling, Informatic Center, Universidade Federal da Paraı´ba, João Pessoa, Paraı´ba, Brazil a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 anawyse@ci.ufpb.br OPEN ACCESS Citation: Wyse AP, Santos AJBd, Azevedo JdS, Lima JSd, Faria JRd (2018) Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal. PLoS ONE 13(10): e0205879. https://doi.org/ 10.1371/journal.pone.0205879 Citation: Wyse AP, Santos AJBd, Azevedo JdS, Lima JSd, Faria JRd (2018) Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal. PLoS ONE 13(10): e0205879. https://doi.org/ 10.1371/journal.pone.0205879 Editor: Baogui Xin, Shandong University of Science and Technology, CHINA Editor: Baogui Xin, Shandong University of Science and Technology, CHINA Copyright: © 2018 Wyse et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Due to recent advances in genetic manipulation, transgenic mosquitoes may be a viable alternative to reduce some diseases. Feasibility conditions are obtained by simulating and analyzing mathematical models that describe the behavior of wild and transgenic popula- tions living in the same geographic area. In this paper, we present a reaction–diffusion model in which the reaction term is a nonlinear function that describes the interaction between wild and transgenic mosquitoes, considering the zygosity, and the diffusive term that represents a nonuniform spatial spreading characterized by a random diffusion parame- ter. The resulting nonlinear system of partial differential equations is numerically solved using the sequential operator splitting technique, combining the finite element method and Runge-Kutta method. This scheme is numerically implemented considering uncertainty in the diffusion parameters of the model. Several scenarios simulating spatial release strate- gies of transgenic mosquitoes are analyzed, demonstrating an intrinsic association between the transgene frequency in the total population and the strategy adopted. Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Competing interests: The authors have declared that no competing interests exist. Malaria is the best-known and most deadly of the vector-borne diseases. According to the World Health Organization [2], in 2016, 91 countries reported a total of 216 million cases of malaria, resulting in 445,000 deaths, approximately the same number as was reported in 2015. However, among the vector-borne diseases, the incidence of dengue has the fastest growth in the world, with a 30-fold increase over the past 50 years. In addition to malaria and dengue, yellow fever, chikungunya, zika, and others are also prominent. Competing interests: The authors have declared that no competing interests exist. Vector control is the primary method to prevent and reduce the transmission of these dis- eases. Combined strategies are strongly recommended to achieve the greatest benefit of con- trolling the proliferation of diseases, minimizing impacts to the ecosystem as well as adverse effects on public health. Therefore, it becomes necessary to understand the local vector ecology and the local patterns of disease transmission so that we can choose the appropriate vector control tools from the wide range of available options. With recent advances in the genetic manipulation of mosquitoes, in particular, Aedes spp. and Anopheles spp., there has been considerable enthusiasm in the search for novel vector con- trol approaches to prevent a wide variety of diseases. In this way, the use of genetically modi- fied mosquitoes can be a promising alternative to control mosquito–borne infections in addition to the traditional prophylaxis, vaccines, medication, and insecticides. The conception of disease control by genetic manipulation is not new; the concept arose in the 1930s-40s for the control of arthropod vectors by three independent research pioneers: A. S. Serebrovskii [3], F. L. Vanderplank [4] and E. F. Knipling [5]. Considerable research on genetic strategies to control mosquitoes was conducted during the 1970s (e.g., see Curtis et.al [6]) on mosquito populations with sterile males. A review of the applications of genetics to vec- tor control can be seen in Refs. [7–9]. The notion of increasing a gene frequency in a vector population so that it interferes with a pathogen has made substantial progress, e.g., Collins and James [10], whose hypothesis resulted in the reduction or elimination of the transmission of that pathogen. Introduction Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Vector-borne diseases have always been of great concern to populations and government authorities in countries with a tropical climate, especially those with a low human develop- ment. The combination of favorable climate and precarious sanitation provides ideal environ- mental conditions for the proliferation of many vectors causing various types of diseases that affect the populations of these regions, leading to considerable economic and social disruption. Due to climate change, intensive farming, dams, irrigation, deforestation, population move- ments, rapid unplanned urbanization, and increases in international travel and trade [1], the incidence of tropical diseases has been extended to regions beyond the tropics, making such diseases a worldwide focus. Funding: This work was partly supported by National Council for Scientific and Technological Development - Brazil, www.cnpq.br, grants 441489/2014-1 awarded to J. S. A. and by Universidade Federal da Paraı´ba. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. 1 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal continuous version was obtained in 2007 [23], and a model considering the impact of trans- genic mosquitoes on malaria transmission was presented in 2011 [24]. In addition, a discrete time model, without the distinction of zygosity and taking into account only the horizontal and vertical transmission of a genetically modified bacterium was proposed by Li [25]; in this case, the horizontal transmission depends on mating between wild mosquitoes and those that have mutated due to contact with the bacteria. Continuous modeling with the distinction of zygosity was proposed by Wyse et al. [26, 27], based on the initial sketch for the interaction between three varieties of mosquitoes: wild, heterozygous and homozygous transgenic [28]. A nondimensional version was considered in 2011, in which the fitness of wild and transgenic mosquitoes was assumed to be distinct [29]. In 2015, Unckless et al. [30] adopted a Wright-Fisher model, with random mating in a pop- ulation of infinite size, to determine the existence and stability of an internal equilibrium when there is a fitness cost to the MCR allele [19], resulting in conditions for successful gene fixation and invasion. In 2018, Noble et al. [21] proposed a stochastic, Moran-based model for evaluat- ing the dissemination efficiency of the self-propagating homing drives in finite wild popula- tions considering the resistant alleles, suggesting its use to suppress wild populations of mosquitoes transmitting malaria. Strategies for delivering malaria-resistant [31, 32] and sterile genetically modified mosqui- toes [33] used techniques of optimal and inundative control, respectively. Studies such as these allow for the achievement of sustainable genetic control programs, help public health agencies in making decisions, and indicate the amount of transgenics required, which genetic composi- tion is most appropriate, and the ideal and local timing for release, as well as cost implications. In this paper, we propose a new mathematical model that describes the dynamics of the interaction by the mating between transgenic and wild-type mosquitoes, as well as the spread- ing of the transgene that determines the interruption of an epidemiological process. For this purpose, the transgenic mosquitoes are differentiated according to their zigozity, being hetero- zygous or homozygous. The interaction between mosquitoes describes a density dependence effect for vital rates and imposes a maximum limit of population growth that occurs according to the carrying capacity. In addition, all populations are in absolute numbers, preserving parameters that can describe effects such as seasonality, stochasticity, etc. The spreading is gov- erned by Fick’s law, with a random diffusion coefficient. Thus, the mathematical model obtained is a nonlinear reaction-diffusion system with uniformly distributed parameters. This resulting system is solved by operator splitting methods that are well known in problem solv- ing that result in large systems of partial differential equations, as well as problems involving nonlinear chemical reactions [34], mosquito dispersal [35], nonlinear Schrodinger applica- tions [36] and others. Sequential and iterative splitting for nonlinear problems with locally Lipschitz-continuous operators, which generates the lowest splitting error and convergence of first order, was presented and verified in [37]. We use the sequential operator to split the sys- tem into two parts and solve each one separately with specialized numerical techniques, in this case, combining the well-known fourth order Runge-Kutta method for the nonlinear dynamic problem and the space-time finite element method with distributed parameters for the diffu- sion problem. The numerical solution obtained, from different initial conditions, show some of the possible scenarios to be discussed to investigate the spreading of transgenic mosquitoes, their behavior with uncertainty parameters and guidelines for future investigations. Laboratory-based discovery research is now underway on a variety of genetic engineering strategies for controlling arthropod vectors of disease, producing mosquitoes that, when released, reproduce with wild mosquitoes and cause their offspring to be refractory to diseases [11, 12], unable to detect CO2 [13], carrying a lethal gene [14], sterile [9] or infected by Wolba- chia pipientis, an endosymbiotic bacteria able to inhibit viral replication in the mosquito [15] and to reduce its lifespan [16]. Genetically modified mosquitoes refractory to malaria, for example, were first obtained in 2002 using a technique developed by Catteruccia et al. [17]. To obtain them, scientists have developed two different types of transgenic mosquitoes of species Anopheles stephensi using the CP (carboxypeptidase) promoter: one of them expresses the syn- thetic peptide SM1 (salivary gland and midgut binding peptide 1) [11], while the other expresses the enzyme PLA2 (phospholipase A2) present in bee venom [12]. In 2010, Amenya et al. [18] used this technique to obtain A. stephensi transgenic lines containing ϕC31 attP “docking” sites linked to a fluorescent marker gene. More recently, in 2015, Gants and Bier [19] developed a method based on the self-propagating CRISPR/Cas9 genome editing technol- ogy, which converts heterozygous to homozygous mutations. This artificial drive mechanism, called a mutagenic chain reaction (MCR), was tested for Drosophila melanogaster with a 97% efficiency. Next, MCR inheritance proved to be efficient to A. stephensi, although lines obtained by mating between drive-males and wild-type females presented a higher drive effi- ciency than lines obtained by mating between drive-females and wild-type males [20, 21]. Assuming these ideas and linking them with mathematical models, the dynamics governing the interaction between wild and transgenic mosquitoes has been subsequently studied. For example, in 2004, Li presented a discrete system model that considered the interaction between two varieties of mosquitoes: wild and transgenic, without the distinction of zygosity [22]; a PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 2 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal on strategies aimed at the genetically modified mosquitoes that are engineered to have a reduced transmission capacity of a particular pathogen and are fertile and able to propagate and perpetuate their inheritable trait in the wild mosquito population. For this, we consider transgenic mosquitoes of the genus Anopheles, more precisely A. stephensi. Studies have shown a reduction in the fitness of this mosquito in relation to the wild one when homozygous trans- genic lines were used [38] and no significant effect when the heterozygous lines were used [18, 39, 40]. One of the known factors that can result in reduced fitness is inbreeding depression associated with the rearing of transgenic mosquitoes as homozygotes. A. stephensi is the key vector of malaria in urban areas of India, a country responsible for six percent of the world’s malaria cases in 2016 (216 million confirmed cases), according to the World Malaria Report 2017 [2]. In many Indian towns, this mosquito has becomes truly domestic [41]. There is increasing evidence of the adaptation of anophelines to organically polluted water bodies [42]. Studies show preferences to deep water as a breeding habitat [43] but also a diver- sity of habitats, with activity in fresh or salt water marshes, mangrove swamps, rice fields, grassy ditches, the edges of streams and rivers and small, temporary rain pools, which can be the result of human behaviors such as broken water pipes, cans, poorly maintained drains, overhead tanks, truck tire tracks on unpaved roads, and low lying areas that are liable to flood- ing among others [44–46]. Based on this information, the mathematical model will assume the simplified hypothesis that females are able to find a breeding in any place of the domain con- sidered, obtaining equal reproductive success. This makes sense in urban areas, for example, where the incidence of malaria is evident [41]. The dispersal distance of mosquitoes in urban areas is considerably less than in open envi- ronments, with the local topography being one of the several extrinsic and intrinsic factors influencing the dispersal of mosquitos. Different values for the dispersal of mosquitoes can be found in the literature, varying due to meteorological conditions, topography, physiological status of released mosquitoes, body size, population density, availability of oviposition and resting sites. Studies conducted by Bailey et al. [47] concluded that the short distance dispersal of Culex tarsalis was independent of wind direction, whereas longer distance dispersal (> 5 km) was always downwind. This finding gives us guidelines of the possibility of this fact occur- ring for other mosquito species (Diptera: Culicidae). Being that the premise of the proposed model is based on urban mosquito dispersal, no advection will be considered. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Materials and methods This section describes population dynamics resulting from the introduction of genetically modified mosquitoes into wild-type populations. The proposed mathematical model is based 3 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Mathematical modeling of the mosquito dispersal Assuming a stable environment, competition for resources may occur in immature P forms, implying losses in adulthood [49] and leading to density-dependent mortality gP ui C , C with γ being the intrinsic rate of growth, C the carrying capacity, and ∑ui = u1 + u2 + u3, or nat- ural death δ. These processes account for the dynamics of each subgroup over time. Since allele combinations are equally likely to occur, the matings are random and a Punnett Square can be a tool to predict the probability of a mating producing each genotype [50]. Because mosquitoes are diploid organisms, it is proper to establish a wild mosquito’s genotype as (w, w) and homo- zygous transgenic genotype as (g, g). After crossing, we established homozygous populations (w, w) and (g, g) and a heterozygous population (w, g), as seen in Table 1. The dominance of w or g defines the phenotype of the heterozygous mosquitoes and depends on genetic manipula- tion, but it is outside the scope of this paper. Let aij, bij and cij denote genotypic frequencies for u1, u2 and u3 obtained from mating ui × uj, i, j = 1, 2, 3. These coefficients must satisfy the relation aij + bij + cij = 1. From the above assumptions, the dynamics of interactions between wild and transgenic mosquitoes with spreading can be established by the following spatiotemporal model with uncertainty parameters: Let (O; F; P) be a probability space in which the variations of the random inputs exhibit, where O is the set of outcomes, F 2O is the σ-algebra of events and P : F ! ½0; 1 is a prob- ability measure. Mathematical modeling of the mosquito dispersal Let ui, i = 1, 2, 3, denote populations of wild, heterozygous and homozygous transgenic mos- quitoes. If we assume that the movement of individual mosquitoes is similar to Brownian motion in a nonuniform field, then we can define the rate of change of the mosquito density at time t 2 Īt (0, T], T > 0, as @uiðx; tÞ @t ¼ ki @2uiðx; tÞ @x2 x 2 I d; ð1Þ ð1Þ where I d ¼ ½0; L IR is the spatial domain, with L > 0, κi denotes the random diffusion rate coefficients of both wild and transgenic mosquitoes. These coefficients aim to evolve a proba- bilistic response, creating a random heterogeneous medium. We also assume that the bound- ary of region Īd does not permit mosquito transport, and thus, we have zero flux boundary conditions @ui @x ð0; tÞ ¼ @ui @x ðL; tÞ ¼ 0: ð2Þ ð2Þ PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 4 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Table 1. Genotypic offspring frequencies obtained from the mating between wild, heterozygous and homozygous transgenic mosquitoes. Genotype Mating u1 × u1 u1 × u2 u1 × u3 u2 × u2 u2 × u3 u3 × u3 (w, w) a11 a12 a13 a22 a23 a33 (w, g) b11 b12 b13 b22 b23 b33 (g, g) c11 c12 c13 c22 c23 c33 https://doi.org/10.1371/journal.pone.0205879.t001 Table 1. Genotypic offspring frequencies obtained from the mating between wild, heterozygous and homozygous transgenic mosquitoes. https://doi.org/10.1371/journal.pone.0205879.t001 This environment affecting mosquitoes’ dispersal behavior was considered by Lutambi et al. [48], and the change in mosquito density was described by a discrete-space continuous-time mathematical model. Additionally, Dufourda and Dumont [35] showed that environmental parameters, such as vegetation, may have an intrinsic influence on mosquito distribution and the efficiency of vector control tools. Taking into account the relationship between environ- mental variability and the dispersal of mosquitoes, it is feasible to consider diffusion coeffi- cients that are not deterministic parameters. The mathematical model assumes that populations ui have the same fitness, according to studies about A. stephensi [39], ensuring equal reproductive success and reflecting how well transgenic mosquitoes are adapted to the wild environment, with the ability to compete and mate. Each individual progresses and emerges into adulthood at a rate and leaves due to mor- tality. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 with Neumann boundary conditions given by th Neumann boundary conditions given by @ui @x ð0; tÞ ¼ @ui @x ðL; tÞ ¼ 0; ð4Þ ð4Þ and initial conditions and initial conditions uiðx; 0Þ ¼ uiðxÞ; ð5Þ ð5Þ Here the diffusion coefficients κi are uniformly distributed random fields in the interval ½si; sf IR, written as κi * U(si, sf). These coefficients are used to improve the computational model and to calibrate a known parameter in order to build a regularized solution ui. Here the diffusion coefficients κi are uniformly distributed random fields in the interval ½ Here the diffusion coefficients κi are uniformly distributed random fields in the interval ½si; sf IR, written as κi * U(si, sf). These coefficients are used to improve the computational model and to calibrate a known parameter in order to build a regularized solution ui. Disregarding the diffusion and adding up the three eq (3), the total mosquito population is described by the Verhulst–Pearl logistic equation with harvesting dN dt ¼ gN 1 N C d2N; ð6Þ ð6Þ where N = ∑ui is the total population of mosquitoes and γ = −δ1, where is the rate of the emergence of mosquitoes into adulthood, and δ1 is the mortality rate due to natural causes. The harvesting effect represented by the density-independent mortality rate δ2 ensures the sta- bilization of the population below the carrying capacity C, more precisely, at the nonnull equi- librium point where N = ∑ui is the total population of mosquitoes and γ = −δ1, where is the rate of the emergence of mosquitoes into adulthood, and δ1 is the mortality rate due to natural causes. The harvesting effect represented by the density-independent mortality rate δ2 ensures the sta- bilization of the population below the carrying capacity C, more precisely, at the nonnull equi- librium point The harvesting effect represented by the density-independent mortality rate δ2 ensures the sta- bilization of the population below the carrying capacity C, more precisely, at the nonnull equi- librium point N ¼ C 1 d2 g : ð7Þ ð7Þ The mosquitoes’ population dynamics described by Eq (6) was fitted to field data for some regions [28, 51] with accurate results. Mathematical models considering variable mosquito populations were used in [52, 53] and [51], suggesting the logistic form to an intraspecific competition. with Neumann boundary conditions given by Considering diffusion in Eq (6), we obtain the Fisher-Kolmogorov model with harvesting. For results that guarantee the model governed by system (3) without dispersal is biologically and mathematically well-posed, see S1 Appendix. Mathematical modeling of the mosquito dispersal Given faij; bij; cij; ; g; C; dg 2 IR, the model problem is to find the population of mosquitoes uiðx; t; wÞ 2 IR for all x 2 Īd, t 2 Īt and ω 2 O that satisfy the following system of nonlinear boundary value problem with random diffusion coefficients @u1 @t ¼ k1ðoÞ @2u1 @x2 þ P ui g C X X aijuiuj du1; @u2 @t ¼ k2ðoÞ @2u2 @x2 þ P ui g C X X bijuiuj du2; @u3 @t ¼ k3ðoÞ @2u3 @x2 þ P ui g C X X cijuiuj du3; 8 > > > > > > > > > > < > > > > > > > > > > : ð3Þ ð3Þ PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 5 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 5 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal with Neumann boundary conditions given by with Neumann boundary conditions given by PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Numerical Formulation In this section, we focus on the development of discrete formulations and applications of computational techniques to numerically solve the proposed model. For this purpose, we pres- ent an operator splitting method [54] used to solve the nonlinear reaction-diffusion system (3). The idea is to decouple the original system into another equivalent system, generating a combination of two distinct physical subsystems formed by problems of less complexity. The advantage of adopting this type of strategy is that we can prevent systems that require costly operations and have the possibility to choose the most optimized algorithms for each operator. Let us consider the sequential operator-splitting method, which solves two subproblems sequentially on each time interval and connects them via the initial conditions. To describe the developed algorithm, we proceed with a natural decomposition of the system (3) into two cou- pled new problems: a system of partial differential equations with the purely diffusive term and a system of nonlinear differential equations associated with the purely reactive term. To solve the diffusive problem, we use the semidiscrete finite element method with an implicit finite difference scheme of the Crank-Nicolson type. The second system, which is nonlinear, is solved using the fourth-order Runge-Kutta method for its high accuracy. The uncertainty on the diffusion parameter is quantified using the Monte Carlo method. 6 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Introducing the temporal discretization I t ¼ ½0; T ¼ SN n¼0 Itn, such that Itn ¼ ½tn; tnþ1 is a partition of It, and N = T/Δt is the number of partitions of It with an uniform time step Δt = tn+1 −tn, we follow the procedure below: Step 1: For the initial time, t = t0, initialize the variables ui(x, t0) = ūi(x) for each i = 1, 2, 3. Step 2: For some fixed n 0, considering known the initial conditions ui(x, tn) and defining ŭi(tn) = ui(x, tn), we calculate ũi(x, t) in the time step tn+1 through the following problem: Step 2: For some fixed n 0, considering known the initial conditions ui(x, tn) and defining ŭi(tn) = ui(x, tn), we calculate ũi(x, t) in the time step tn+1 through the following problem: Problem A: Given κi: O ! PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Numerical Formulation (0, 1), to find ~uiðx; t; oÞ 2 IR, with x 2 Īd, t 2 Itn and ω 2 O, satis- fying the following system: @~uiðx; t; oÞ @t ¼ kiðoÞ @2~uiðx; t; oÞ @x2 ; ð8Þ ð8Þ with boundary conditions with boundary conditions ð~uiÞxð0; tÞ ¼ ð~uiÞxðL; tÞ ¼ 0; ð9Þ ð9Þ and initial conditions and initial conditions ~uiðx; tnÞ ¼ uiðtnÞ: ð10Þ ð10Þ Step 3: In the same time interval Itn, we use the solution of Problem A, given by the previous step, as the initial condition to calculate the solution of the system of coupled nonlinear ordinary differential equations, associated with the model (3), expressed by the following problem: Problem B: Given faij; bij; cij; g; C; ε; dg 2 IR, find uiðtÞ 2 IR, t 2 Itn satisfying the following system: du1 dt ¼ P ui g C P P aijuiuj du1; du2 dt ¼ P ui g C P P bijuiuj du2 du3 dt ¼ P ui g C P P cijuiuj du3; 8 > > > > > > > > > > < > > > > > > > > > > : ð11Þ ð11Þ with initial condition with initial condition uiðtnÞ ¼ ~uiðx; tnþ1Þ; ð12Þ ð12Þ where ũi(x, tn+1) are the solutions obtained from Problem A. where ũi(x, tn+1) are the solutions obtained from Problem A. Step 4: The solution of Problem B is given by the approximate solution of the model at tnþ1 2 Itn It. If tn+1 < T, we increment n, return to step 2, and repeat the process until equality occurs. To solve Problem A, we adopted a finite element method with uniformly distributed parameters using a Monte Carlo scheme to obtain the first and second moment of the solution. The spatial domain is discretized using an uniform partition Ie with ne elements, such that Id ¼ Sne e¼1 Ie and Tne e¼1 Ie ¼ ;. Numerical Formulation The method seeks an approximation ~uh i ðx; t; xðkÞÞ to ~uiðx; t; oÞ in PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 7 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal the Lagrangian finite element space of class C0(Id) and degree l given by Vh ¼ fvhðx; tÞ 2 C0ðIdÞ j vhð0Þ ¼ vhðLÞ ¼ 0; 8 vhðIeÞ 2 ^PlðIeÞg. (k) Vh ¼ fvhðx; tÞ 2 C0ðIdÞ j vhð0Þ ¼ vhðLÞ ¼ 0; 8 vhðIeÞ 2 ^PlðIeÞg. It is defined by requiring that, for each realization on the diffusion parameter {κi(ξ(k)) 2 [κmin, κmax]} (k = 1, . . ., Ns) and for all vh 2 Vh, Z Id @~uh i vh @t dx þ Z Id kiðxðkÞÞ @~uh i @x @vh @x dx ¼ 0: ð13Þ ð13Þ Concerning Problem B given by the ODE system Eq (11), we use the classical fourth-order Runge-Kutta method to obtain its solution. Concerning Problem B given by the ODE system Eq (11), we use the classical fourth-order Runge-Kutta method to obtain its solution. Finally, the statistical moments of the discrete variables are computed by generating a popu- lation of Ns samples f~un i ð; xðkÞÞ; k ¼ 1; . . . ; Ns; i ¼ 1; 2; 3g corresponding to the problems A and B. The mean and variance are computed using the approximations mð~uiÞ ¼ 1 Ns X Ns k¼1 ~uið; xðkÞÞ ð14Þ ð14Þ s2ð~uiÞ ¼ 1 Ns X Ns k¼1 ð^uið; xðkÞÞÞ 2; ð15Þ ð15Þ respectively, where ûi = ûi −μ(ũi) is the fluctuation around the mean. respectively, where ûi = ûi −μ(ũi) is the fluctuation around the mean. respectively, where ûi = ûi −μ(ũi) is the fluctuation around the mean. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Results and Discussion Parameter Description Value κi Diffusion coefficients(Km2/week) [0.01, 0.05] Emergence rate to the adult stage (week−1) 5.1 δ1 Density-dependent death rate (week−1) 3.9 δ2 Density-independent death rate (week−1) 0.5 C Carrying capacity 2, 500 https://doi.org/10.1371/journal.pone.0205879.t002 https://doi.org/10.1371/journal.pone.0205879.t002 include the uncertainty in a scenario where the diffusion of wild and transgenic mosquitoes varies randomly in the same range. The random solution of the model was computed by the Monte Carlo method, employing 150 unconditional realizations generated from a uniform distribution in the interval [si, sf] and considering the diffusion coefficient κi, i = 1, 2, 3 on this interval. The quality of these realizations was examined for each proposed experiment of the model by comparing the statistical sample’s mean and the cumulative variance of these realizations. In this paper, we are considering an average sex ratio approximated as 1:1, as is usually done. Estimates on the finite rate of increase for A. stephensi were obtained by Suleman [56] in laboratory conditions resulting in 1.19 (the population multiplied 1.19 times per day). With this information we can evaluate the weekly finite rate of increase as (1.19)7 = 3.38, and then the weekly intrinsic rate of increase is γ = ln(3.38) = 1.2. We estimated the instantaneous emer- gence-rate using the life table from Suleman [56] and expressions from [57], resulting in a rate of 5.1 per week and a death rate δ1 = (5.1 −1.2) = 3.9 per week. Obviously, 1.2 is a theoreti- cal potential to increase, and in nature, the population is subject to predation and human action inducing a density-independent mortality, which we will assume to be δ2 = 0.5 per week. The carrying capacity was assumed to be 2,500 mosquitoes. The female dispersal (y) for A. stephensi was adjusted by a linear function of the release time (x), y = 58.741 + 23.731x, by Reisen and Aslamkhan [58], resulting in a weekly dispersal of 224.86m. Therefore, the diffusion coefficient can be obtained from k ¼ <x2> qit , with < x2 > the mean square displacement and qi a numerical constante related to dimensionality (qi = 2, 4, or 6, for 1, 2, or 3-dimensional diffusion, respectively), resulting in κ = 0.0253km2 per week. Thus, it is reasonable to consider the diffusion coefficient varying in the interval [0.01, 0.05]. All parameters used in numerical simulations are shown in Tables 2 and 3. Results and Discussion In this section, we report the results obtained when applying computational techniques to numerically solve the proposed diffusion-reaction model, which is an analysis of the behavior of the three mosquito varieties in different scenarios, considering the mendelian (as SM1 [12] mutation, for example) and MCR inheritance [20]. Five situations are considered for numeri- cal experiments. The first one attempts to establish a correlation with a controlled laboratory experience described by Moreira et al. [12]. The following experiments establish different mos- quito release scenarios to establish the most effective release strategies. For this purpose, we simulate the population dynamics of each population with different initial conditions. For the mendelian inheritance, no homozygous transgenic mosquito is released, but its population arises naturally from the mating process. In laboratory experiments, it is common to obtain homozygous lines through successive crosses between heterozygous mosquitoes; thus, these mosquitoes are not subject to inbreeding, which can lead to a reduction in fitness [38, 55]. For the MCR inheritance, mosquitoes of the three genotypes will be released, respecting the suc- cess rate of the MCR technique in relation to the Mendelian inheritance because of the initial conditions. Indeed, we assume that there is no flow of mosquito populations on the boundary. In all experiments, we consider that mosquitoes occupy a spatial region Īd = [0, 30], mea- sured in km, whose spreading starts from the release of the mosquitoes and has a duration of twelve weeks, Īt = [0, 12], corresponding to one season of the year, to avoid seasonal changes in the model parameters. The finite element mesh adopted consists of three intervals with a uniform mesh sequence. The first and last intervals, given by [0, 10] and [20, 30], respectively, have meshes with a num- ber of nodes (n node) equal to 20. In the central region, between ]10, 20[, we consider a more refined mesh with 40 nodes. Linear interpolations (two nodes elements) and quadratic inter- polations (three nodes elements) were considered. As there is no evidence of differences in the flight capacity for wild and transgenic mosqui- toes, all mosquitoes have the same probability of dispersal. For this reason, these experiments PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 8 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Table 2. Parameters of model estimated from literature data [56, 58]. Simulations using the initial conditions of the laboratory experience In this experiment, we compared the Hardy-Weinberg frequencies obtained in the laboratory experiment conducted by Moreira et al. [39] with the numerical simulation of the spatial-tem- poral model (3) considering the spatial dynamics and adopting equivalent initial conditions. In their experiment, the authors confined in a cage equal numbers of wild and heterozygous transgenic mosquitoes, the latter characterized by encoding the anti-Plasmodium SM1 gene in the gut and expressing an enhanced GFP (Green Fluorescent Protein) marker in the eye. After five generations, there was a frequency of 44% of positive GFP and 56% negative GFP, which is consistent with the Hardy-Weinberg equilibrium for allele frequencies p = 0.75 and q = 0.25; thus, they do not imply fitness load. The numerical simulation was obtained considering a population composed of 1,000 wild and 1,000 heterozygous transgenic mosquitoes, homogeneously spread over 4 km, according to the following initial condition: u1ðx; 0Þ ¼ u2ðx; 0Þ ¼ ( 250 if 13 x 17; 0 if 0 x < 13 and 17 < x 30 u3ðx; 0Þ ¼ 0: ð16Þ ð16Þ u3ðx; 0Þ ¼ 0: This result means that the transgene frequency in the parental population was q = 1/4, as in the laboratory experiment, and invariant throughout the 4 km, ensuring that encounters between mosquito varieties occurred with the same probability and randomness. To check the quality of the realizations, we computed the results of various samples in the last step of time and calculated the relative error of the mean and variance of wild mosquitoes defined as ErelðNsÞ ¼ k mu1ðNsÞ mu1ðNs þ 1Þ k k mu1ðNs þ 1Þ k ; ð17Þ ð17Þ where mu1ðNsÞ denotes the mean or variance of Ns samples generated by the target solution which was computed on L2-norm. Once the relative error of the mean and variance for the Nr realizations were computed, we chose the target solution with 150 realizations since the fluctu- ation of the relative error in both moments converged to values close to or below 1% (see Fig 1). Similar results were obtained for the transgenic populations, and therefore, they were omit- ted from the text in all other experiments. In Fig 2 we plotted the behavior of the mean and cumulative variance of the three mosquito varieties. Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal frequency a22 = 0.25; heterozygous transgenic u2 are generated from the mating u1 × u2 with frequency b12 = 0.5, u2 × u2 with frequency b22 = 0.5, u2 × u3 with frequency b23 = 0.5 and u1 × u3 with frequency b13 = 1; homozygous transgenic u3 are generated from the mating u2 × u2 with frequency c22 = 0.25, u2 × u3 with frequency c23 = 0.5, and u3 × u3 with frequency c33 = 1. Considering the MCR inheritance, heterozygous transgenic mosquitoes are converted to homozygous transgenic mosquitoes with a sucess rate f, being 1-f the resistance to this conversion. The efficiency converting heterozygous A. stephensi to homozygous transgenic mosquitoes was 98% when obtained from the mating between transgenic males and wild females and 14% when obtained from mating between wild males and transgenic females [20, 21]. In these sim- ulations we will consider f = 0.14 (MCR) and f = 0 (Mendelian inheritance). Results and Discussion According to the mendelian inheritance, wild mosquitoes u1 are generated from the mating u1 × u1 with frequency a11 = 1, while u1 × u2 with frequency a12 = 0.5 and u2 × u2 with Table 3. Genotypic frequencies considering Mendelian and MCR inheritance. Frequency a11 a12 a13 a22 a23 a33 Mendelian 1.0 0.5 0.0 0.25 0.0 0.0 MCR 1.0 0.5 0.0 0.25 0.0 0.0 b11 b12 b13 b22 b23 b33 Mendelian 0.0 0.5 1.0 0.5 0.5 0.0 MCR 0.0 0.5(1-f) 1.0(1-f) 0.5(1-f) 0.5(1-f) 0.0 c11 c12 c13 c22 c23 c33 Mendelian 0 0.0 0.0 0.25 0.5 1.0 MCR 0 0.5f 1.0f 0.25+0.5f 0.5+0.5f 1.0 htt //d i /10 1371/j l 0205879 t003 Table 3. Genotypic frequencies considering Mendelian and MCR inheritance. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 9 / 25 Simulations using the initial conditions of the laboratory experience The average number of mosquitoes obtained at the end of each quarter, represented by mean μT(), that resulted from the integration of the obtained solution into the Īd domain PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 10 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 1. Statistical convergence of the first and second moment of the wild mosquito population. Relative error: (a) Mean and (b) Variance. https://doi.org/10.1371/journal.pone.0205879.g001 Fig 1. Statistical convergence of the first and second moment of the wild mosquito population. Relative error: (a) Mean and (b) Variance. https://doi.org/10.1371/journal.pone.0205879.g001 https://doi.org/10.1371/journal.pone.0205879.g001 using the trapezoidal rule is presented in Table 4. In the second moment, we used the cumula- tive variance in order to better visualize the discrepancy between the average number of mos- quitoes for each variety obtained from the fixed and random diffusion parameters. Table 4 shows that the means obtained at the end of the integration process correspond to approximately 56% wild mosquitoes and 44% transgenic (heterozygous plus homozygous) mosquitoes, which is in agreement with the experiment described in [39]. Moreover, these proportions are in accordance with the Hardy-Weinberg equilibrium for ŭ1(0) = ŭ2(0) = 0.5 and ŭ3(0) = 0 (see S1 Appendix), which are equivalent to the initial conditions (16). We also note that the cumulative variance in all experiments starts very small due to the strong influence of the deterministic initial data. As time goes by, the profile grows with a higher uncertainty in that there is a growth near 10 km, and then, the population stabilizes between 10 km and 20 km, expanding again after that limit and then stabilizing. At the end of the simula- tion process, which comprises a 12-week period, the cumulative variance of the total number of mosquitoes was approximately 68% wild mosquitos and 32% transgenic mosquitoes. Simulations involving effective study of mosquito release strategies In the following experiments, we describe scenarios obtained from the numerical simulation of model (3) for different initial configurations of mosquito distribution in the domain. In all PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 11 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 2. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the population of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (16). https://doi.org/10.1371/journal.pone.0205879.g002 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 2. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the population of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (16). https://doi.org/10.1371/journal.pone.0205879.g002 cases, we consider an initial population composed of 2,000 wild, 1,000 heterozygous and no homozygous mosquitos for the mendelian case and 2,000 wild, 860 heterozygous and 140 homozygous mosquitoes for the MCR case, distributed over 30 km in different ways. These values were assumed considering that 14% of the heterozygous mosquitoes became cases, we consider an initial population composed of 2,000 wild, 1,000 heterozygous and no homozygous mosquitos for the mendelian case and 2,000 wild, 860 heterozygous and 140 homozygous mosquitoes for the MCR case, distributed over 30 km in different ways. These values were assumed considering that 14% of the heterozygous mosquitoes became Table 4. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (16). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 2.9986e+03 4.4260e+03 5.6266e+03 6.8316e+03 μT(u2) 1.9990e+03 2.9507e+03 3.7511e+03 4.5544e+03 μT(u3) 333.1725 491.7788 625.1760 759.0633 htt //d i /10 1371/j l 0205879 t004 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 12 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal homozygous due to the MCR technology. The analysis of these simulations gives us informa- tion about the best way to release transgenic mosquitoes, aiming to increase in this population the frequency of the gene that interferes with the development of the protozoan. Simulations involving effective study of mosquito release strategies For this simu- lation, we assumed that wild mosquitoes are in a region classified as one of high incidence and that they spread to a region of lower concentration within a domain, being contained due to the interaction with transgenic mosquitoes, which spread in the same way. Experiment 1-Mendelian: In this experiment, we consider an initial population distributed over 30 km in the following way: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 250 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; ð18Þ ð18Þ u3ðx; 0Þ ¼ 0: u3ðx; 0Þ ¼ 0: u3ðx; 0Þ ¼ 0: u3ðx; 0Þ ¼ 0: These conditions characterize a release in which 2,000 wild mosquitoes are homogeneously distributed in nature, strewn over a 4km strip. In this simulation, 1,000 heterozygous trans- genic mosquitoes are introduced, maintaining the same uniformity characteristic of wild mos- quitoes. This distribution characterizes a transgene frequency in the parental population of q = 1/6, along this strip. This allele frequency (p = 5/6, q = 1/6) leads to a Hardy–Weinberg equilibrium of f(w, w) = 0.6944, f(w, g) = 0.2778 and f(g, g) = 0.0278. Fig 3 illustrates the means and cumulative variances of wild, heterozygous and homozygous transgenic mosquitoes. As the release form of this experiment is similar to the previously dis- cussed laboratory experience, it is expected that the behavior of the graphs in these figures have the same evolution at the statistical moments found in Fig 2. The experimental results are summarized and listed in Table 5. The increase in wild mosquitoes in the initial conditions passed from the ratio of 1:1 in (16) to 2:1 in (18) led, at the end of simulation, to proportions of 69.44% wild, 27.78% het- erozygous and 2.78% homozygous transgenic mosquitoes, with a frequency of transgenic alleles of q = 0.167. These values are expected and consistent with the Hardy-Weinberg equi- librium for the initial conditions of proportions ŭ1(0) = 2/3, ŭ2(0) = 1/3 and ŭ3 (0) = 0, which are equivalent to the initial conditions in (18). This initial distribution consequently contributed to the cumulative variance growth of wild mosquitoes and the decrease in trans- genic mosquitoes. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Under these conditions, 2,000 wild mosquitoes are homogeneously distributed in nature, strewn over a 4 km strip. In the same region, 860 heterozygous and 140 homozygous Simulations involving effective study of mosquito release strategies Experiment 1-MCR: In this experiment, we consider an initial population distributed over 30 km in the following way: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 215 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u3ðx; 0Þ ¼ ( 35 if 13 x 17; 0 if 0 x < 13 and 17 < x 30: ð19Þ ð19Þ Under these conditions, 2,000 wild mosquitoes are homogeneously distributed in nature, strewn over a 4 km strip. In the same region, 860 heterozygous and 140 homozygous PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 13 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 3. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (18). https://doi.org/10.1371/journal.pone.0205879.g003 Fig 3. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (18). https://doi.org/10.1371/journal.pone.0205879.g003 https://doi.org/10.1371/journal.pone.0205879.g003 transgenic mosquitoes are introduced maintaining the same uniformity characteristic of wild mosquitoes. The frequency of transgenic alleles in the parental population is q = 0.19. The evo- lution of the three population for 12 weeks can be seen in Fig 4. From the mean, we can see a high population growth of transgenic mosquitoes, pre-empting the homozygous mosquitoes. transgenic mosquitoes are introduced maintaining the same uniformity characteristic of wild mosquitoes. The frequency of transgenic alleles in the parental population is q = 0.19. The evo- lution of the three population for 12 weeks can be seen in Fig 4. From the mean, we can see a high population growth of transgenic mosquitoes, pre-empting the homozygous mosquitoes. Table 5. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (18). Simulations involving effective study of mosquito release strategies This situation is described by initial conditions: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 1; 000 if 14:5 x 15:5; 0 if 0 x < 14:5 and 15:5 < x 30; u3ðx; 0Þ ¼ 0: ð20Þ ð20Þ u3ðx; 0Þ ¼ 0: u3ðx; 0Þ ¼ 0: Here, populations are not evenly distributed in the same space, and therefore, mating occurs naturally among the nearest mosquito varieties. Due to this distance-induced prefer- ence, mating is not random, and the Hardy-Weinberg equilibrium cannot be verified. This deviation of randomness by geographic dispersion will occur in all experiments hereafter. Numerical simulations corresponding to the first and second moments using initial condi- tions (20) are shown in Fig 5. We can also observe that the growth in the cumulative variance is more pronounced in heterozygous and homozygous transgenic mosquitoes. In the early part of the simulation, in intervals [13, 14.5] and [15.5, 17] almost all the mating and generating are by wild mosquitoes; therefore, wild-type mosquitoes cause the increase in the mean of this variety in these intervals. In the region [14.5, 15.5], there are twice as many transgenic mosquitoes compared to wild ones, with the mating generating the three varieties of mosquitoes. Gradually peaks and valleys tend soften and merge. After twelve weeks, we obtain from Table 7 proportions of 89.63% wild, 9.57% heterozygous and 0.8% homozygous trans- genic mosquitoes, with a final transgene frequency of q = 0.056, much lower than the Hardy- Weinberg frequency of q = 0.167 obtained in Experiment 1—Mendelian. Comparing these val- ues obtained using (20) with those obtained from the initial conditions (18), we can observe an increase in wild mosquitoes and a decrease in transgenic mosquitoes due to the method of release. Releasing transgenic mosquitoes in a more restricted region induced 27.36% more wild mosquitoes and 66% and 71.63% fewer heterozygous and homozygous transgenic mosquitoes, respectively, than in a homogeneous release process over 4 km. Simulations involving effective study of mosquito release strategies Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 4.1832e+03 5.6913e+03 7.1280e+03 8.6228e+03 μT(u2) 1.6733e+03 2.2765e+03 2.8512e+03 3.4491e+03 μT(u3) 167.3289 227.6532 285.1201 344.9122 https://doi.org/10.1371/journal.pone.0205879.t005 estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (18). PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 14 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 4. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (19). https://doi.org/10.1371/journal.pone.0205879.g004 Fig 4. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (19). https://doi.org/10.1371/journal.pone.0205879.g004 https://doi.org/10.1371/journal.pone.0205879.g004 Table 6 shows the total populations at 3, 6, 9 and 12 weeks. After the final week, 99.62% of population was composed of homozygous transgenic mosquitoes, and 0.37% was composed of heterozygous mosquitoes, implying a frequency of transgenic alleles of q = 0.998. The propor- tion of the wild type of mosquito was not relevant. Table 6. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (19). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 1.0439e+03 76.9052 2.5391 0.0690 μT(u2) 2.5367e+03 1.2388e+03 273.2076 49.7368 μT(u3) 2.5418e+03 7.1854e+03 1.0538e+04 1.3175e+04 https://doi.org/10.1371/journal.pone.0205879.t006 Table 6. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (19). PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 15 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Experiment 2-Mendelian: In this experiment, we simulate the mosquitoes’ release consid- Experiment 2-Mendelian: In this experiment, we simulate the mosquitoes’ release consid- ering the same initial distribution of wild mosquitoes as the previous experiment; however, we adopt the release of 1,000 heterozygous transgenic mosquitoes in a concentrated way, restricted to a small region of 1 km. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Simulations involving effective study of mosquito release strategies Experiment 2-MCR: In this experiment, we adopted the same release strategy as the men- delian case, but considering initial conditions for MCR: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 860 if 14:5 x 15:5; 0 if 0 x < 14:5 and 15:5 < x 30; u3ðx; 0Þ ¼ ( 140 if 14:5 x 15:5; 0 if 0 x < 14:5 and 15:5 < x 30: ð21Þ ð21Þ Fig 6 shows the mean and cumulative variance of the three populations at four moments. Due to initial conditions, there is a strong decay of the wild mosquito population at the center of the domain, contrasting with the homozygous mosquito population which continues to grow as time progresses. Furthermore, the population of heterozygous mosquitoes has a strong PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 16 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 5. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (20). https://doi.org/10.1371/journal.pone.0205879.g005 Fig 5. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (20). https://doi.org/10.1371/journal.pone.0205879.g005 https://doi.org/10.1371/journal.pone.0205879.g005 growth in the intervals [9.5, 14.5] and [15.5, 21]. The cumulative variances indicates a balance in the population growth for both trangenic and wild mosquitoes. growth in the intervals [9.5, 14.5] and [15.5, 21]. The cumulative variances indicates a balance in the population growth for both trangenic and wild mosquitoes. Finally, 12,821 mosquitoes were accounted. The final proportions were 16.02% wild type, 18.23% heterozygous and 65.75% homozygous transgenic, as shown in Table 8. A release of Table 7. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (20). Simulations involving effective study of mosquito release strategies Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 4.6029e+03 6.7495e+03 8.8277e+03 1.0982e+04 μT(u2) 952.7485 1.0881e+03 1.1433e+03 1.1727e+03 μT(u3) 140.8560 123.4126 108.7569 97.8511 https://doi.org/10.1371/journal.pone.0205879.t007 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 17 / 25 Table 7. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (20). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 4.6029e+03 6.7495e+03 8.8277e+03 1.0982e+04 μT(u2) 952.7485 1.0881e+03 1.1433e+03 1.1727e+03 μT(u3) 140.8560 123.4126 108.7569 97.8511 https://doi.org/10.1371/journal.pone.0205879.t007 estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (20). Table 7. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using 17 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 6. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (21). https://doi.org/10.1371/journal.pone.0205879.g006 Fig 6. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of the populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (21). https://doi.org/10.1371/journal.pone.0205879.g006 https://doi.org/10.1371/journal.pone.0205879.g006 transgenic mosquitoes according to Experiment 2 (Mendelian or MCR) is the more intuitive manner of inserting transgenic mosquitoes into the environment; however, this method is the least effective. Despite this result, the experiment showed that the MCR is efficient even in this situation, leading to a final frequency of transgenic alleles of q = 0.75. Table 8. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (21). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 3.1396e+03 3.2292e+03 2.8612e+03 2.0545e+03 μT(u2) 1.3376e+03 1.8159e+03 2.1513e+03 2.3371e+03 μT(u3) 1.2706e+03 3.1068e+03 5.4399e+03 8.4295e+03 https://doi.org/10.1371/journal.pone.0205879.t008 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 18 / 25 Table 8. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (21). Table 8. Simulations involving effective study of mosquito release strategies Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (21). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 3.1396e+03 3.2292e+03 2.8612e+03 2.0545e+03 μT(u2) 1.3376e+03 1.8159e+03 2.1513e+03 2.3371e+03 μT(u3) 1.2706e+03 3.1068e+03 5.4399e+03 8.4295e+03 https://doi.org/10.1371/journal.pone.0205879.t008 18 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Experiment 3-Mendelian: In this experiment, we investigate the dynamics of the three spe- cies of mosquitoes, considering a release region of the heterozygous transgenic mosquitoes larger than the natural region of the wild-type mosquitoes. For this, we consider an initial con- dition in which 2,000 wild mosquitoes are found in a 4 km region, and 1,000 heterozygous transgenic mosquitoes are evenly distributed in a 6 km region. This initial configuration is given by the following conditions: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 166:67 if 12 x 18; 0 if 0 x < 12 and 18 < x 30; ð22Þ ð22Þ u3ðx; 0Þ ¼ 0: u3ðx; 0Þ ¼ 0: To begin, only heterozygous transgenic mosquitoes are over the intervals [12, 13] and [17, 18]. Mating between mosquitoes of this population generates 50% heterozygous, twice the other varieties; because of this, there is an increase in heterozygous transgenic mosquitoes in these regions. Mating between heterozygous mosquitoes also generates more homozygous transgenic mosquitoes than mating between mosquitoes of a mixed population composed of the wild type and the heterozygous; thus, the population of homozygous transgenic mosqui- toes is also increased in positions [12, 13] and [17, 18]. This dynamic is shown in Fig 7, which illustrates the mean and cumulative variance of the three populations at four moments. Table 9 shows that the growth of wild mosquitoes is lower than that presented in previous experiments, comprising 43.7% of total population. On the other hand, the numbers of heterozygous and homozygous transgenic mosquitoes showed a very significant growth, reaching 40% and 16.3% of the total population, respectively. Consequently, the population growth rate in the cumulative variation is higher for transgenic mosquitoes. PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Simulations involving effective study of mosquito release strategies Therefore, the release of transgenic mosquitoes that extrapolate the focus region of the wild-type mosquitoes presents the best way of transgenic mosquito insertion into the total population, resulting in a frequency of the transgene alleles of q = 0.363, the highest value obtained from Mendelian genetics. Experiment 3-MCR: This initial configuration is given by the following conditions: u1ðx; 0Þ ¼ ( 500 if 13 x 17; 0 if 0 x < 13 and 17 < x 30; u2ðx; 0Þ ¼ ( 143:33 if 12 x 18; 0 if 0 x < 12 and 18 < x 30; ð23Þ ð23Þ u3ðx; 0Þ ¼ ( 23:33 if 12 x 18; 0 if 0 x < 12 and 18 < x 30: considering the same pattern of initial distribution as the previous experiment, but with values considering the same pattern of initial distribution as the previous experiment, but with values considering the same pattern of initial distribution as the previous experiment, but with values PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 19 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 7. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (22). https://doi.org/10.1371/journal.pone.0205879.g007 Fig 7. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (22). https://doi.org/10.1371/journal.pone.0205879.g007 https://doi.org/10.1371/journal.pone.0205879.g007 according to the MCR. We plot the mean and cumulative variance corresponding to these ini- tial conditions (see Fig 8). Fig 8 exhibits the dominance of the homozygous mosquito population and the decay of the other populations. This is reflected in the cumulative variance that retains a strong Table 9. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (22). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 4.2924e+03 5.4036e+03 5.9782e+03 6.4293e+03 μT(u2) 2.2320e+03 3.5904e+03 4.8033e+03 5.9805e+03 μT(u3) 506.5637 1.0281e+03 1.6238e+03 2.2700e+03 https://doi.org/10.1371/journal.pone.0205879.t009 Table 9. Conclusion In this paper, we present the development of a new mathematical model to describe the inter- action and spreading of wild and transgenic mosquitoes taking into account the zygosity, con- sidering all populations in absolute numbers, and admitting random diffusion coefficients. These characteristics make the model description more realistic, ensuring the existence of all possible phenotypes, preserving the parameters that do not exist in dimensionless form and allowing for the mosquitos a nonuniform displacement. The dynamic system was developed to preserve the peculiarities of the species and avoid an overlap of individuals when the transgenics are inserted, respecting the environmental support capacity. The terms describing mating and competition imply the nonlinearities of this system, which are characteristic of the great majority of population dynamics models. The diffusion term, based on Fick’s law, describes a symmetrical spread of the mosquito population with a random dispersal. The solution of the proposed problem was obtained using the operator splitting method to decouple the diffusion-reaction system into two subproblems, in which the diffusive problem was solved employing the finite element method and the reactive problem using the fourth order Runge-Kutta method. The results shown are consistent with the expected behavior, indicating a constant presence of transgenic mosquitoes after entering the ecosystem and reducing periodic insertion costs, as is the case with sterile mosquitoes. Numerical simulations provide guidelines, indicating a higher efficiency when the release of the transgenics is done in a broader coverage. On the other hand, the worst strategy is the release of transgenic mosquitoes concentrated in a small single region, allowing for the wild type to mate and breed, while the contact with transgenics is not yet accomplished. Without the imposition of transgenics’ superiority, the total elimina- tion of wild-type mosquitoes is impossible to achieve, since they are also obtained from the mating between heterozygous transgenics. It is worth mentioning that the extinction of a spe- cies should not be the main objective; it is sufficient that the population of wild-type mosqui- toes is reduced to levels that do not harm human health, and this is possible for both classical Mendelian genetics and MCR. However, the frequency of transgenic alleles for the MCR is much higher than that for Mendelian genetics, MCR being the most efficient technique in all the experiments presented. Simulations involving effective study of mosquito release strategies Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (22). Table 9. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Ca simulations and initial conditions (22). https://doi.org/10.1371/journal.pone.0205879.t009 20 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Fig 8. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (23). https://doi.org/10.1371/journal.pone.0205879.g008 Fig 8. Display of the moments at times T = 3, 6, 9, 12 weeks of (a)-(c)-(e) mean and (b)-(d)-(f) cumulative variance of populations of wild (a-b), heterozygous (c-d) and homozygous transgenic mosquitoes (e-f) with initial conditions (23). https://doi.org/10.1371/journal.pone.0205879.g008 https://doi.org/10.1371/journal.pone.0205879.g008 variability in homozygous mosquitoes while the other population has an almost negligible variability. y Table 10 shows that of the 14,434 mosquitoes obtained at the end of the 12 weeks, 99.7% were homozygous transgenic and 0.3% were heterozygous, indicating that the transgenic allele is the most frequent, reaching a frequency of q = 0.9985. Table 10. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (22). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 1.2716e+03 97.0692 2.9900 0.0753 μT(u2) 2.6539e+03 1.3061e+03 264.8893 44.2056 μT(u3) 3.0904e+03 8.5488e+03 1.1985e+04 1.4390e+04 https://doi.org/10.1371/journal.pone.0205879.t010 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 21 / 25 Table 10. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (22). Mean Time T = 3 T = 6 T = 9 T = 12 μT(u1) 1.2716e+03 97.0692 2.9900 0.0753 μT(u2) 2.6539e+03 1.3061e+03 264.8893 44.2056 μT(u3) 3.0904e+03 8.5488e+03 1.1985e+04 1.4390e+04 https://doi.org/10.1371/journal.pone.0205879.t010 Table 10. Total populations of the corresponding estimated values of the mean at T = 3, 6, 9, 12, using Monte Carlo simulations and initial conditions (22). 21 / 25 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Conclusion The impact of reducing infectious diseases transmitted by mosquitoes can be studied by coupling the model proposed in this paper with an appropriate epidemiological dynamic. By identifying an effective breeding site, it is possible to plan an optimal strategy for the release of transgenic mosquitoes, optimizing the allocation of scarce resources. Thus, the entry of geneti- cally modified individuals into the ecosystem can be a viable alternative to disease control, as long as they are able to interact with the wild mosquitoes and have at least the same survival capacity in the environmental field. This work is an initial proposal that opens possibilities for future investigations. Initial con- ditions most appropriate to reality need to be tested in order to obtain a strategy for the release of transgenic insects into the environment. Factors such as seasonality, temperature, winds, rainfall, and fitness cost can be introduced into this model, or in a two-dimensional version of it, making it even more realistic. The choice of the finite element method to solve the diffusive problem is intended to facilitate some of these future steps. Author Contributions Conceptualization: Ana Paula Wyse, Josenildo Silva de Lima. Conceptualization: Ana Paula Wyse, Josenildo Silva de Lima. Data curation: Ana Paula Wyse, Jairo Rocha de Faria. Formal analysis: Ana Paula Wyse, Antonio Jose´ Boness dos Santos, Juarez dos Santos Aze- vedo, Jairo Rocha de Faria. Funding acquisition: Ana Paula Wyse, Juarez dos Santos Azevedo. Investigation: Ana Paula Wyse, Josenildo Silva de Lima. Methodology: Ana Paula Wyse, Antonio Jose´ Boness dos Santos, Juarez dos Santos Azevedo, Josenildo Silva de Lima, Jairo Rocha de Faria. Software: Antonio Jose´ Boness dos Santos, Juarez dos Santos Azevedo. Validation: Josenildo Silva de Lima. Visualization: Antonio Jose´ Boness dos Santos, Juarez dos Santos Azevedo, Josenildo Silva de Lima. Writing – original draft: Ana Paula Wyse, Antonio Jose´ Boness dos Santos, Juarez dos Santos Azevedo, Jairo Rocha de Faria. Supporting information S1 Appendix. (PDF) S1 Appendix. (PDF) 22 / 25 PLOS ONE \| https://doi.org/10.1371/journal.pone.0205879 October 31, 2018 Modeling the spreading and interaction between wild and transgenic mosquitoes with a random dispersal Author Contributions Conceptualization: Ana Paula Wyse, Josenildo Silva de Lima. Data curation: Ana Paula Wyse, Jairo Rocha de Faria. Formal analysis: Ana Paula Wyse, Antonio Jose´ Boness dos Santos, Juarez dos Santos Aze- vedo, Jairo Rocha de Faria. 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https://openalex.org/W2780152486_2	Spanish-Science-Pile	Open Science	Various open science	2,017	Crítica de la economía biopolítica: dimensiones políticas del pensamiento médico / Criticism of the biopolitical Economics: political dimensions of medical thinking	None	Spanish	Spoken	6,334	11,153	Por tanto, al hacer una genealogía de cómo se configura de modo contemporáneo esa crítica al Estado como un ente intrínsecamente expansivo y totalizador, Foucault muestra cómo esta arraiga en el diagnóstico que los neoliberales efectúan del Welfare State como una forma de “intervencionismo estatal” que, situándola al lado de los estados fascistas y comunistas, constituiría una amenaza que es necesario contrarrestar. A partir de esas reflexiones, el ordoliberalismo alemán desplegará una reformulación de las relaciones entre sociedad civil y Estado que postulaba el liberalismo. Allí donde el liberalismo concebía el mercado como un espacio natural de intercambio ante el cual era necesario laissez faire -no intervenir- el neoliberalismo lo concebirá como un espacio KAMCHATKA 10 (DICIEMBRE 2017): 13-38 26 Ester Jordana Lluch. Crítica de la economía biopolítica… de competencia permanente cuyo dinamismo hay que animar. Paso, pues, de una “sociedad mercancía” a una “sociedad empresa” donde el gobierno de la sociedad se concebirá como el gobierno de las relaciones entre individuos que se conducen como “empresarios de sí mismos”. En el marco de esas transformaciones, Foucault dará cuenta de cómo las políticas neoliberales abordarán esas problemáticas relaciones entre el Estado y la “seguridad social” y la “seguridad criminal”. Para ello analiza, por un lado, las propuestas de Giscard en Francia y, por otro, ciertas reformulaciones de la tecnología penal desplegadas en Estados Unidos señalando cómo ambas se manifiestan como la extensión – y radicalización- de ese modelo ordoliberal que caracteriza nuestras sociedades contemporáneas. En el caso Francés, Giscard, retomando las propuestas de Stoléru y Stoffaes postulaba la posibilidad de un “impuesto negativo” (2007b, 242) concebido como un subsidio que podría ser asignado a aquellos sujetos en “riesgo de exclusión” incapaces de proveerse por sí mismos todo un conjunto de servicios como la educación o la sanidad. Por tanto, la propuesta para minimizar todo el sistema de “seguridad social” se basaría en fijar un umbral en función del cual se estableciese qué sujetos deben optar a esos subsidios mientras el resto de la población debería proveérselos por sí mismos. Un umbral que, por tanto, configura un nuevo modo de exclusión/inclusión que podrá ser establecido políticamente de modo oscilante según las necesidades del mercado. Lejos de las viejas aspiraciones al pleno empleo acompañaban las políticas del Welfare State, esas políticas neoliberales garantizarían más bien una suerte de “existencia mínima” (2007b, 248) a todo un conjunto de la población sin necesidad de que trabajen. Por tanto, en el marco de esa forma de sociedad empresa formulada por el ordoliberalismo alemán, esas medidas constituirían un modo de eliminar la cuestión de la “seguridad social” del tablero, haciéndola pasar por completo del lado del mercado: Ahora bien, un proyecto semejante no es otra cosa que la radicalización de los temas generales acerca de los cuales les hablé con referencia al ordoliberalismo, cuando los ordoliberales alemanes explicaban que el objetivo principal de una política social no era, por cierto, hacerse cargo de todos los riesgos que pudiesen afectar a la masa global de la población, y agregaban que una verdadera política social debía ser tal que, sin tocar en absoluto el juego económico y, por consiguiente, dejando que la sociedad se desarrollara como una sociedad de empresa, se estableciera una serie de mecanismos de intervención para asistir a quienes lo necesitaran en el momento , y sólo en el momento que lo necesitaran (Foucault, 2007b, 248). En el seno del neoliberalismo americano, Foucault constataba un desplazamiento similar en torno a la relación entre crimen y seguridad. A partir de los argumentos de Stiegler Foucault mostrará cómo el crimen, analizado en el seno de esa racionalidad securitaria, dejaría de ser contemplado como algo que hay que eliminar para ser concebido como un fenómeno que es necesario regular subordinando esa intervención a un cálculo de costes. Concibiendo el crimen, no ya en términos antropológicos, sino como una “acción” ante la cual un individuo corre el riesgo de ser “castigado”, podrá desplazarse la pregunta por “quién es” el sujeto susceptible de castigo por la de “cualquiera” que cometa una acción susceptible de punición o de responder a ciertas intervenciones cuyo objetivo sería disuasorio y calibrable en términos de ganancias o pérdidas. A partir de ese desplazamiento, la KAMCHATKA 10 (DICIEMBRE 2017): 13-38 27 Ester Jordana Lluch. Crítica de la economía biopolítica… ley será susceptible de reinscribirse como uno más de los múltiples mecanismos disuasorios y reguladores que conforman esa lógica securitaria para funcionar dentro de esa gubernamentalidad ambiental. En otras palabras, todas las distinciones antes establecidas, todas las distinciones que pudieron introducirse entre criminales natos, criminales ocasionales, perversos y no perversos, reincidentes, etc., no tienen ninguna importancia. Es preciso admitir que, de todas formas, por patológico, si se quiere, que sea el sujeto en determinado nivel y visto desde determinada perspectiva, hasta cierto punto, en cierta medida es responsive a los cambios en las ganancias y las pérdidas; vale decir que la acción penal debe ser una acción sobre el juego de las ganancias y las pérdidas posibles, una acción ambiental (Foucault, 2007b, 302). A raíz de todos los ámbitos analizados hasta ahora podemos retomar cómo a la luz de las hipótesis desarrolladas en los cursos de Seguridad, Territorio, Población y El nacimiento de la biopolítica cabe resituar ese arco general de transformación en torno a las diferentes relaciones entre medicina y Estado en el siglo XIX y el siglo XX. Esa mutación marca también la diferencia entre el diagnóstico que Foucault efectúa en torno al Plan Beveridge y el Welfare State a través las críticas que los neoliberales realizarán respecto al mismo. Como hemos visto, en el marco del neoliberalismo la tendencia sería hacer desaparecer todos aquellos mecanismos disciplinarios que, en el siglo XIX, se habían articulado con esa formado parte de esa necesidad de “defender la sociedad” de todo un conjunto de peligros con que Foucault caracterizaba la relación entre población y Estado en el marco de liberalismo. Ahora bien, esas críticas neoliberales eludían, a través de esa crítica del Estado, la antinomia intrínseca al propio liberalismo que está en la base de esa pendiente: el conflicto que, en el marco de ese gobierno biopolítico de la población, induce la relación siempre problemática de una “exposición” al peligro y las estrategias de “seguridad” desplegadas ante el mismo. Por tanto, ese modelo ordoliberal constituye un nuevo modo de gubernamentalidad que no deja de intensificar, de otro modo, la tendencia que ya introducía el liberalismo: una sociedad donde la “seguridad” (y con ella, la regulación del “orden social”) están “por encima de la ley” (Foucault, 2001d, 366). En una entrevista realizada en 1977, Foucault señalaba cómo, a diferencia del “pacto territorial” característico del modelo soberano, lo que caracteriza nuestras sociedades es, pues, un “pacto de seguridad”. Y esa relación implica que “es un Estado que está obligado a intervenir en todos los casos en que un acontecimiento singular, excepcional, perfora la trama de la vida cotidiana”. (20012, 50) que irá de la mano, no de una suerte de regulación totalitaria ni efectuada a partir de un exceso de poder sino, dirá Foucault, de una respuesta solícita en tanto que se presenta como dispuesto a intervenir, incluso más allá de la ley, ante cualquier acontecimiento extraordinario que amenace esa seguridad. De ahí, afirmaba, la amenaza y la angustia que suscita el terrorismo en tanto que pone en tela de juicio esa capacidad real de garantizar la seguridad de la población. Por tanto, por un lado, nos encontraríamos ante un sistema altamente “insecurizante” que, al mismo tiempo, expone nuestras vidas a todo un conjunto de intervenciones, controles, vigilancias, regulaciones, en nombre de esa seguridad: KAMCHATKA 10 (DICIEMBRE 2017): 13-38 28 Ester Jordana Lluch. Crítica de la economía biopolítica… Ya no nos encontramos en el orden de los accidentes cubiertos por la sociedad "aseguradora"; estamos en presencia de una acción política que "inseguriza" no solo la vida de los individuos, sino la relación de estos con todas las instituciones que hasta el momento los protegían. De ahí la angustia provocada por el terrorismo. Angustia en los gobernantes. Angustia también en las personas que otorgan su adhesión al Estado, aceptan todo, los impuestos, la jerarquía, la obediencia, porque el Estado protege y garantiza contra la inseguridad (Foucault, 2012, 51). Por otro, añadía, se trata de un tipo de poder mucho más sutil y hábil que el que habían puesto en marcha los sistemas totalitarios en tanto que va acompañado, como veíamos, de una aparente tolerancia a determinados fenómenos y comportamientos que, sin embargo, se permiten mientras estos no devengan “peligrosos” en el marco de esa regulación biopolítica general: Las sociedades de seguridad que están en proceso de formación toleran por su parte toda una serie de comportamientos diferentes, variados, en última instancia desviados y hasta antagónicos entre sí; con la condición, es cierto, de que se inscriban dentro de cierto marco que elimine casas, personas y comportamientos considerados como accidentales y peligrosos. Esta delimitación del "accidente peligroso" corresponde efectivamente al poder. Pero en ese marco se toleran un margen de maniobra y un pluralismo infinitamente más grandes que en los totalitarismos. Es un poder más hábil, más sutil que el del totalitarismo. (Foucault, 2012, 51). Retomando la paradoja a la que apuntaba Foucault entre esos planes estatales y la movilización bélica de la población la pregunta es, pues, qué sucederá ahora que ese “pacto de seguridad” fracasa en tanto que el Estado deja de ser el garante securitario para desplazar todo un conjunto de ámbitos de los que se hacía cargo a las dinámicas del mercado. Por otro lado, en el marco bélico, poco a poco los “servicios militares” y la consideración de la población civil como población susceptible de ser llamada a filas ha desaparecido. Por tanto, ya no existe esa gubernamentalidad que, construida en torno a la Razón de Estado, hacía del mismo un espacio al mismo tiempo de obediencia y salvación. En segundo lugar, en esa mutación, el papel del Estado en relación a la sanidad y el fomento de una “economía de la salud” cambia sustancialmente en tanto que los Estados neoliberales, a diferencia del Welfare State, inscriben en el ámbito económico no solo la “salud” en términos amplios sino las propias infraestructuras de acceso a los cuidados y las curas. Ese desplazamiento constituye una mutación fundamental del modo en que la población es “expuesta” a los riesgos que, como veíamos, constituían una relación inherente al propio liberalismo. Así, enlazando una y otra problemática, se preguntaba Foucault: Un aparato fabricado para asegurar la seguridad de las gentes en el terreno de la salud ha alcanzado pues un punto de su desarrollo en el que va a ser preciso decidir qué-determinada enfermedad, qué determinado tipo de sufrimiento ya no se beneficiará de ninguna cobertura —un punto en el que la vida misma, en determinados casos, no recibirá ninguna protección—. Esto plantea un problema político y moral que se asemeja, guardando las diferencias, a la cuestión de saber qué derecho tiene un Estado para exigir a un individuo que vaya a dejarse matar en la guerra. Esta cuestión, sin haber perdido en absoluto nada de su carácter acuciante, ha sido perfectamente integrada en la conciencia de la gente a través de largos derroteros históricos de tal forma que los soldados han efectivamente aceptado el hacerse KAMCHATKA 10 (DICIEMBRE 2017): 13-38 29 Ester Jordana Lluch. Crítica de la economía biopolítica… matar —y por tanto situar su vida al margen de la protección—. La cuestión que se suscita en el presente es saber cómo las personas van a aceptar estar expuestas a determinados riesgos sin conservar el beneficio de una cobertura por el Estado-providencia (Foucault, 1985, 222) Por tanto, si en el marco del Welfare State la pregunta política era hasta qué punto el Estado podía o no satisfacer unas demandas potencialmente infinitas en el marco de la salud y cómo era el encargado de regular y decidir qué quedaba cubierto y qué no en función de unos presupuestos estatales (Foucault, 2001d, 1186), la entrada de la salud en el mercado neoliberal donde el Estado se sitúa como una pieza clave de la dinamización del mismo radicaliza esa dimensión económica en tanto que el criterio de decisión pasa a ser la rentabilidad misma. CRÍTICA DE LA ECONOMÍA BIOPOLÍTICA En vistas a todo lo expuesto vamos a finalizar, pues, desplegando algunos de los ejes problematizados por Foucault en torno a la cuestión biopolítica tratando de acompañar de modo simultáneo las mutaciones que el neoliberalismo contemporáneo introduce en los mismos y hacia dónde podría hacerlos bascular. Si sus estudios habían girado en torno a cómo el desarrollo de las ciencias humanas no podía aislarse de una historia económica y política, a nuestro juicio, la pendiente introducida por el neoliberalismo apunta a un desbordamiento de las relaciones que había estudiado Foucault situándonos en una gubernamentalidad que se desplaza más allá de las mismas. Por tanto, y situando esas reflexiones en un marco meramente especulativo, vamos a tratar de extender esas reflexiones foucaultianas hasta los límites de nuestro presente permitiéndonos trazar algunos lineamientos posibles más allá de lo analizado por el autor. En primer lugar, en torno a la problemática que introducía la OMS en relación al efecto de los tratamientos antibióticos y antimicrobianos, Foucault aludía en las citadas conferencias impartidas en Brasil en 1974 a una cuestión que tan solo allí será expuesta explícitamente aunque esté íntimamente relacionada con la cuestión misma de la biopolítica. Se trataba de plantear cómo ciertas intervenciones médicas contemporáneas habrían tenido la potestad de afectar, por primera vez en la historia, al desarrollo de la vida misma. En el momento en que se introducen masivamente esos tratamientos antibióticos y anti-infecciosos, señalaba allí Foucault, no solo se interviene sobre aquellas patologías que tratan de combatir sino que, en términos generales, esa intervención modifica el umbral de sensibilidad de los organismos ante esos agentes. Una problemática que, como vemos, sigue siendo objeto de preocupación en la comunidad médica dado que en unas décadas podríamos encontrarnos ante el hecho de que esos antibióticos ya no tienen efecto en tanto que los virus se han hecho inmunes a los mismos. Lo mismo, señalaba el autor, puede decirse en relación a la manipulación genética de células vivas, de bacilos o virus o de cómo la tecnología médica ha permitido desarrollar armas biológicas para las que no hay defensa y que sigue constituyendo un motivo de alarma ante el desencadenamiento de un conflicto bélico que involucre su uso. Por tanto, en esos aspectos, como vemos, no se trata de que la vida esté expuesta en términos de la falta de saber médico en determinados campos, algo inherente a la historia misma de la medicina, sino de cómo es el KAMCHATKA 10 (DICIEMBRE 2017): 13-38 30 Ester Jordana Lluch. Crítica de la economía biopolítica… despliegue positivo de la misma el que, por primera vez en la historia, introduce una mutación relevante en tanto que la vida misma puede ser modificada. De ahí que Foucault señalase, como apuntábamos, a que hemos entrado en una “biohistoria”: En nuestros días se descubre algo nuevo: la historia del hombre y la vida están profundamente imbricadas entre sí. La historia del hombre no continúa simplemente la vida, ni se limita a reproducirla, sino que la retoma, hasta cierto punto, y puede ejercer sobre su proceso una serie de efectos totalmente fundamentales. En segundo lugar, en relación a la polémica generada en torno al DSM-V, como veíamos, desde el propio espacio médico y psiquiátrico se alertaba de una posible sobrediagnosticación y sobrepatologización de la vida cotidiana acompañada de una sobremedicación. Ahora bien, más allá de esa reflexión crítica del propio campo psiquiátrico en relación a los límites diagnósticos que cabe estipular, la cuestión de esa patologización de la vida cotidiana presenta un problema mucho más profundo. En el seno del ámbito clínico y psiquiátrico la controversia en torno al peso que cabe otorgar a la dimensión psicobiológica y su intervención farmacológica atraviesa todo el siglo XX. Una intensa discusión desarrollada particularmente en torno a la cuestión de la terapéutica. El psicoanálisis, de modo intenso, pero también numerosas corrientes de corte sistémico o humanista, han apostado por defender que el espacio de la clínica terapéutica no puede ser substituido por un tratamiento farmacológico abanderando en muchos casos una lucha en torno a las consecuencias de una medicalización generalizada de la población. Dejando a un lado esa discusión en torno a los límites de lo psíquico o lo biológico en relación a las patologías psiquiátricas y su terapéutica, el problema a plantear es si esa controversia no se despliega ya en unos marcos epistemológicos que hacen imposible atender a la dimensión política y social del malestar. Bajo esa perspectiva, si bien, efectivamente, ese malestar social produce síntoma, abordarlo tan solo en ese extremo reconduce la dimensión histórica y política del mismo a un marco psicobiológico que impide nombrarlo como tal. En el momento en que el hombre es pensado, bien a partir de una naturaleza humana -por definición transhistórica- bien a partir de su historia vital y afectiva concreta, se inhabilita la posibilidad de pensar los modos de transformación histórica en el seno de los cuales se conforman nuestras experiencias. Y ese espacio es, justamente, el que permite abrir la tentativa foucaultiana de pensar al hombre fuera de los límites del pensamiento antropológico. Si bien en el momento en que el malestar social hace síntoma el tratamiento terapéutico (clínico o farmacológico) es lo que permite mantenerse a flote, es necesario dotarse de las herramientas teóricas y prácticas para plantear la dimensión común de ese malestar pudiendo politizar, desde ahí, la crítica de las formas de vida que lo producen. Por otro lado, como veíamos, las reflexiones de Foucault giraban en torno a cómo, en el paso del siglo XIX al siglo XX, todo un conjunto de ámbitos que no estaban “codificados” por la medicina no pueden pensarse hoy fuera de la misma. La higiene, la sexualidad, el cuerpo o la alimentación han pasado a formar parte del ámbito de la “salud”. En relación a la sexualidad, uno de los campos más KAMCHATKA 10 (DICIEMBRE 2017): 13-38 31 Ester Jordana Lluch. Crítica de la economía biopolítica… estudiados por Foucault, el problema permitía plantear, entre otras dimensiones, cómo, cuándo y porqué la sexualidad pudo ser pensada como enfermedad y ser susceptible de intervención médica y “terapéutica” y cuáles son sus consecuencias en el modo de constituirnos como sujetos. Como es sabido, la homosexualidad formaba parte de los primeros manuales diagnósticos psiquiátricos y fue posteriormente eliminada. Si bien ahora esa inclusión forma parte de la historia “incómoda” de la psiquiatría cabe recordar, sin embargo, cómo, pese a la amplia movilización LGTBI, si bien el Trastorno de la identidad sexual ha desaparecido del DSM vigente, permanece el Trastorno de disforia de género. Por tanto, en relación a la intensa movilización en torno a las luchas LGTBI, ese campo de batalla dista de estar zanjado (no hace falta recordar en cuantos países la gente sigue siendo perseguida o censurada por su condición sexual). Ahora bien, en el marco de la relación entre placer, deseo y sexualidad, esa problemática va más allá de su circunscripción clínica. Una parte fundamental de las investigaciones foucaultianas a lo largo de sus últimos años se basaba en explorar cómo a lo largo de nuestra historia se configuró un sujeto de deseo cuyo escrutinio situaría la revelación de una verdad sobre sí mismo. Frente a ello, Foucault exploraba la posibilidad de considerar los placeres como prácticas que, desprovistas de esa relación entre subjetividad y verdad, pudiesen situar en otro lugar esas relaciones. Se trataba de liberar al sujeto de que su identidad, la verdad de quién es, estuviese vinculada a la verdad de su sexualidad. Sin embargo, cabe preguntarse hasta qué punto ese desplazamiento hacia los placeres que Foucault consideraba como emancipatorio de ese dispositivo ha entrado hoy en un nuevo ámbito de gobierno que no es el de esa relación entre el sujeto y su identidad sexual, sino en el de una mercadotecnia del propio placer. Desplazándose del ámbito clínico al ámbito mediático nos encontramos hoy con una intensa producción de esos discursos del "how to do it” vinculados a intensificar, “mejorar” y “cualificar” las prácticas sexuales. Por otro lado, a través de las redes sociales se moviliza intensamente un mercado afectivo y sexual que, lejos de celebrar como una liberación, es necesario problematizar en otros términos ¿Cómo cabe pensar, pues, ese desplazamiento? ¿En qué sentido se sitúan todos esos discursos, prácticas y técnicas en el marco de un gobierno de los afectos, la sexualidad y el placer en sí mismos? Si ante el gobierno normalizado de la sexualidad se produjo en los años sesenta toda una oleada de “liberación” que Foucault analizaba con recelo, nos encontramos ahora con sujetos que más bien se reivindican como “asexuales” (ACES, 2006) apareciendo todo un nuevo tipo de categorías en base a las cuales se abre la posibilidad de “salir” de ese dispositivo sexoafectivo. ¿Cabe, pues, pensar esas manifestaciones como un modo distinto de resistencia ante esa mutación del dispositivo de la sexualidad o se trata de otra cosa? Por otro lado, la cuestión de la alimentación y los efectos derivados de la misma constituyen hoy un campo de intensa problemática social que se despliegan en un nuevo campo de gubernamentalidad biopolítica. El reciente impuesto a las bebidas azucaradas en Cataluña (Generalitat de Cataluña, 2017) no es más que uno de los efectos de superficie de una intensa inquietud en torno a qué comemos, cómo se produce y sus efectos. Y ahí, de nuevo, aparece un nuevo ámbito donde las desigualdades sociales atraviesan la cuestión de la “salud”. Nos adentramos en un nuevo clasismo vinculado a esa “somatocracia” a la que aludía Foucault que, intensificando la salud como un objeto KAMCHATKA 10 (DICIEMBRE 2017): 13-38 32 Ester Jordana Lluch. Crítica de la economía biopolítica… de mercado, despliega todo un nicho en torno a alimentos “saludables” y “ecológicos” -que no están al alcance de todo el mundo- al tiempo que se alerta de las consecuencias para la población del consumo de una “fast food” de bajo coste. Exactamente lo mismo sucede con el amplio mercado a disposición de esa producción del cuerpo que produce todo un conjunto de cuerpos “normativos” como “saludables”, intensificando a su alrededor la exclusión de aquellos cuerpos que no encajan en los mismos. Si en torno a la sexualidad se hacía necesario desplazar esa dimensión del campo de la “salud” en tanto que esa inserción vehiculaba su patologización, ¿qué tipo de relación con el propio cuerpo puede pensarse más allá de ese marco de lo “saludable-no saludable”? Vinculado a esa cuestión de la normalización, como hemos mostrado, esa ha sido una de las cuestiones problematizadas intensamente por Foucault en relación a los mecanismos de clasificación que la hacían posible. Como veíamos, los dispositivos disciplinarios se distinguían de los securitarios, entre otras cosas, por sus modos de funcionamiento en torno a la norma. La distinción binaria que se establecía en torno a lo normal y lo anormal en el modelo disciplinario mutaba, en el modelo securitario, en todo un conjunto de parámetros donde cualquier sujeto es situado en múltiples curvas de normalidad ocupando en ellas posiciones diferentes. El retroceso de las técnicas disciplinarias de la clasificación y el examen de los individuos se traducen hoy en una nueva clasificación transversal de competencias donde la atención se fija en los procesos y en la monitorización o “tutorización” de los mismos. Desplazamiento, por tanto, hacia todo un conjunto de ámbitos donde nuestras capacidades y habilidades pueden ser situadas. No se trata ya de escandir binariamente lo normal de lo anormal sino de situar al sujeto en una amplia grilla de posiciones relacionales que marcarán, como si de una empresa se tratase, sus “Debilidades, Amenazas, Fortalezas, Oportunidades” (Humphrey, 2005). La proliferación de todo un conjunto de estrategias de “gobierno de uno mismo y de los otros” articuladas en torno al coaching, la autoayuda o los manuales de emprendeduría habilitan, pues, un nuevo marco de gubernamentalidad en torno a las conductas, deseos, motivaciones o pensamientos de los sujetos. Situándolos en un permanente juego de inclusión-exclusión de un umbral que debe alcanzar por sí mismo, se trata de autoproducirse permanentemente en base a unos criterios fluctuantes. Por tanto, nuestras vidas ya no están “expuestas” tanto a un sistema de clasificación como a un sistema securitario que nos lleva a calibrar permanentemente nuestros “riesgos”, nuestras “inversiones” o nuestras “capacidades” en el marco de esa gestión empresarial de uno mismo. Por último y, atendiendo a una problemática más específica, Foucault trataba de explorar cómo en el momento en que la dimensión política de la “peligrosidad” social entraba en el campo jurídico, la psiquiatría penal encontró un espacio desde el cual intervenir y dar respuesta. En esa hibridación de una categoría política de “peligrosidad” situada en el marco de una “racionalidad clínica” se abría la posibilidad de criminalizar a los sujetos en base a cómo sus disposiciones, motivaciones o deseos los constituyen en “virtualmente” peligrosos. Foucault problematizaba cómo, en ese proceso, se habría producido una suerte de redoblamiento del sujeto psicológico en torno al sujeto jurídico cuyas consecuencias políticas son que ese dispositivo permita actuar sobre cualquier individuo que KAMCHATKA 10 (DICIEMBRE 2017): 13-38 33 Ester Jordana Lluch. Crítica de la economía biopolítica… considere “peligroso” no ya en base a sus “actos” sino en base a una “peligrosidad” inherente al sujeto mismo, a lo que “es”: Y es que la penalidad moderna -de forma clara y neta a partir de Beccaria- no concede derecho a la sociedad sobre los individuos más que en razón de lo que hacen: un único acto, definido como infracción por la ley, puede dar lugar a una sanción, sin duda modificable en función de las circunstancias o de las intenciones. Pero al poner cada vez más de relieve al criminal como sujeto del acto y también al individuo peligroso como virtualidad de actos, en realidad, ¿no se concede a la sociedad derecho sobre el individuo a partir de lo que él es? (Foucault, 1996, 117). Ahora bien, el problema contemporáneo que plantean de modo intenso dos cuestiones dispares y, sin embargo, intensamente problemáticas como son la del terrorismo y los crímenes machistas, pone en crisis ese dispositivo en tanto que, por un lado, es imposible atravesar esa “peligrosidad” a través de una grilla clínica y, en ambos casos, se plantea el dilema jurídico de actuar antes de que produzca el acto criminal en sí. Si bien en relación a los crímenes machistas vemos cómo más o menos el dispositivo cínico-terapéutico sí tiene herramientas para redescribirlos en términos de “baja tolerancia a la frustración” o “personalidad dependiente o agresiva” no deja de situarse en una posición liminar en tanto que ninguno de esos factores puede explicar per se por qué esa violencia se ejerce sobre las mujeres. Un artículo publicado por El País recientemente en torno a una investigación realizada por la Secretaría de Estado de Seguridad del Ministerio del Interior (El País, 2017) levantaba la polémica (eldiario.es, 2017; Público, 2017) en base a esa redescripción de la violencia machista en términos meramente clínicos. Por otro lado, el caso del terrorismo es aún más peliagudo en tanto que es evidente que es inútil aplicar cualquier tipo de grilla clínica y, sin embargo, el problema penal es palmario ¿cuáles son los límites que permiten actuar sobre un sujeto en base a determinar su “peligrosidad” y “riesgo” antes de cometer cualquier “acto criminal”? ¿Cómo va a determinarse y en base a qué? ¿Qué sistemas de control y vigilancia son necesarios para un sistema que ya no pretende “castigar” los actos cometidos sino evitarlos? Esos dos ejemplos ponen de manifiesto cómo no es el “fracaso” de las ciencias humanas lo que está en juego en su capacidad de determinar la “peligrosidad” de determinados individuos sino más bien la crisis del modo en que históricamente la psiquiatría entró a jugar parte en ese juego de la peligrosidad y la seguridad. Nos encontramos, pues, ante los límites de ese desdoblamiento de lo clínico sobre lo criminal que permitía interrogar los “motivos”, “intenciones”, “deseos” y “pensamientos” de los sujetos en tanto que, a la luz de la dimensión política que atraviesan de modo distinto los dos ejemplos expuestos, resulta difícil que puedan abordarse desde la perspectiva una “historia clínica” individual. KAMCHATKA 10 (DICIEMBRE 2017): 13-38 34 Ester Jordana Lluch. Crítica de la economía biopolítica… Por otro lado, en el seno de sus análisis en torno al neoliberalismo americano Foucault se hacía eco, como veíamos, de cómo este abría un marco de análisis que desbordaba de otro modo esa dimensión antropológica al resituar la conducta criminal como una conducta económica susceptible de ser gobernada en base a un cálculo de costes y beneficios. Desde esa perspectiva se abre una nueva dimensión crítica del sistema jurídico en tanto que el funcionamiento de la ley se inscribe como una dimensión más de esas prácticas gubernamentales que podían regular ciertas conductas a partir de introducir mecanismos disuasorios. La alarmante aplicación de la denominada “Ley mordaza” constituiría un ejemplo de ese desplazamiento en que la ley pasa a formar parte de esa tecnología gubernamental. Como muestra de lo ilimitado de esa aplicación, recientemente aparecía en la prensa cómo vía Facebook un joven había sido multado por mostrar una foto en la red en que aparecía con una camiseta que incluía las siglas A.C.A.B (eldiario.es, 2017). En base a todo lo expuesto, hemos tratado de mostrar las herramientas que Foucault nos brindaba para pensar todo ese amplio conjunto de problemáticas que hibrida la cuestión de la biopolítica con la medicina y la salud tratando de llevarlos más allá de sí mismos. Como vemos, todo un conjunto de problemas que es inútil plantear desde una reducción en base una dicotómica posición a favor o en contra de la medicina, en ningún caso se trata de eso. Podemos valernos como reflexión final del modo en que el autor abordaba la cuestión en relación a las críticas a la “antimedicina” desplegadas por Iván Illich. En alusión a las mismas, Foucault señalaba que no se trata de criticar la medicina en sí misma sino de interrogar el modelo histórico en el que se ha configurado “con el fin de saber en qué medida es posible modificarlo” (Foucault, 2002,360). La cuestión era más bien señalar cómo en el momento en que ese gesto se efectúa se hace en nombre de un cuerpo, una higiene y una salud ya codificadas. Asimismo, cuando la “antipsiquiatría” contraponía a la psiquiatría una “desmedicalización” de la enfermedad, lo hacía también desde un discurso elaborado en base a un saber médico y psiquiátrico. O cuando el peritaje criminal consideraba susceptible de examen a cualquier delincuente, lo que estaba poniendo en tela de juicio no es la responsabilidad penal del acto, sino el grado de peligrosidad social del sujeto medida en términos patológicos (Foucault, 2001d, 52). Del mismo modo, en el caso del Welfare State se trataba de cuestionar, no el derecho a la atención médica, sino el modo en que, queriendo corregir todo un conjunto de desigualdades las producía de otro modo; cómo a través del mismo se planteaba la cuestión de regular políticamente qué enfermedades quedaban cubiertas y cuales no; señalar cómo se relacionaba problemáticamente con el mercado farmacéutico. El ataque neoliberal a sus infraestructuras nos lleva ahora a situar todos esos problemas en una nueva dimensión en tanto que, en el momento en que la salud y las curas médicas pasan del lado del mercado, es este el que va a convertirse en el marco regulador de todas esas dimensiones radicalizándose sus consecuencias políticas. Por tanto, el objeto de la crítica no es cuestionar ese “derecho” a la salud y a la cura, sino preguntarse cómo este puede efectuarse sin todo ese despliegue de tecnologías gubernamentales que lo acompaña y sus efectos. Para Foucault, como veíamos, se trataba de poder desarticular, a partir de esas genealogías cruzadas entre la historia de la medicina como campo de saber y las formas de gubernamentalidad que se hibridan a la misma, el modo en que, de modo contingente, se constituyó KAMCHATKA 10 (DICIEMBRE 2017): 13-38 35 Ester Jordana Lluch. Crítica de la economía biopolítica… históricamente un “pensamiento médico” que desbordó por completo los límites de la propia medicina al funcionar en términos políticos y económicos. Mostrar su carácter contingente es sinónimo de afirmar que es posible pensar un modo de organizar una atención médica no pase necesariamente por mismos los raíles que la configuraron en la historia europea moderna. Así, una “crítica de la economía biopolítica”, nos llevaría, por un lado, a cuestionar esos límites siempre a riesgo de ser desbordados de una patologización de la sociedad y su hipermedicamentalización o los efectos generales de que la “salud” se haya convertido en un campo que despliega en su seno el marco de relación que establecemos con nosotros mismos, con nuestro cuerpo, con nuestra alimentación o con nuestra sexualidad y sus efectos normalizadores. Por otro, se trata de combatir un modo de gubernamentalidad que, bajo la rúbrica de la “libertad” y bajo la antinomia de exponernos doblemente al “peligro” y las tecnologías de “seguridad”, no deja de extender su campo de dominio sobre todos los aspectos de la vida, de quiénes somos, de lo que debemos y podemos esperar de la misma y del modo de conducirla. Los principios axiomáticos del liberalismo y el neoliberalismo según los cuales la maximización del beneficio de cada cual generan un bien general no solo son impugnables, pues, desde una “crítica de la economía política” que, desde al marxismo, refutaba el principio liberal del trabajo como una relación contractual para mostrar cómo en el seno del capitalismo este se convierte en una mercancía más que vehicula la explotación. También lo son desde una “crítica de la economía biopolítica” que permite mostrar cómo el liberalismo y neoliberalismo entendidos como un modo de gubernamentalidad constituyen modos de relación social donde la gestión de la vida no deja de incrementarse. KAMCHATKA 10 (DICIEMBRE 2017): 13-38 36 Ester Jordana Lluch. Crítica de la economía biopolítica… BIBLIOGRAFÍA ACEs (2006). “Acerca de ACEs”. BUTLER, Judith (2017). Cuerpos aliados y lucha política. Hacia una teoría performativa de la asamblea. Barcelona: Paidós. CANGUILHEM, Georges (1986). 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FOUCAULT, Michel (2008). Vigilar y castigar: Nacimiento de la prisión. Traducción de Aurelio Garzón del Camino. Madrid: Siglo Veintiuno. FOUCAULT, Michel (2006a). Historia de la Sexualidad. Vol. 1: La Voluntad de saber. Traducción de Ulises Guiñazú. Madrid: Siglo Veintiuno. FOUCAULT, Michel (2013). La société punitive. Cours au Collège de France (1972-1973). Bernard E. Harcourt, François Ewald y Alessandro Fontana, eds. Paris: Seuil;Gallimard. FOUCAULT, Michel (2007a). El poder psiquiátrico. Curso en el Collège de France (1973-1974). Traducción de Horacio Pons. Buenos Aires: FCE. FOUCAULT, Michel (2001b). Defender la sociedad. Curso en el Collège de France (1975-1976). Traducción de Horacio Pons. Buenos Aires: FCE. FOUCAULT, Michel (2006b). Seguridad, territorio, población: Curso en el Collège de France (1977-1978). Traducción de Horacio Pons. Buenos Aires: FCE. FOUCAULT, Michel (2007b). El nacimiento de la biopolítica. Curso en el Collège de France (1978-1979). Traducción de Horacio Pons. Buenos Aires: FCE. FOUCAULT, Michel (2014a). El gobierno de los vivos. Curso en el Collège de France (1979-1980). Traducción de Horacio Pons. Buenos Aires: FCE. KAMCHATKA 10 (DICIEMBRE 2017): 13-38 37 Ester Jordana Lluch. Crítica de la economía biopolítica… FOUCAULT, Michel (2005). Hermenéutica del sujeto. Curso del Collège de France (1981-1982). Traducción de Horacio Pons. Madrid: Akal. FOUCAULT, Michel (2014b). Obrar mal, decir la verdad. La función de la confesión en la justicia. Curso de Lovaina. Traducción de Horacio Pons. Buenos Aires: Siglo XXI, 2014b. FOUCAULT, Michel (2001c). Dits et écrits I. 1954-1975. Daniel Defert y François Ewald, eds. con la col. de Jacques Lagrange. Paris: Gallimard. FOUCAULT, Michel (2001d). Dits et écrits II. 1976-1988. Daniel Defert y François Ewald, eds. con la col. de Jacques Lagrange. Paris: Gallimard. FOUCAULT, Michel (2002) Obras esenciales II. Estrategias del poder. Traducción de Julia Varela y Fernando Álvarez-Uría. Barcelona: Paidós. FOUCAULT, Michel (2012) El poder una bestia magnífica. Sobre el poder, la prisión y la vida. Traducción de Horacio Pons. Buenos Aires: Siglo XXI. FOUCAULT, Michel (2013) La inquietud por la verdad. Escritos sobre la sexualidad y el sujeto. Traducción de Horacio Pons. Buenos Aires: Siglo XXI. FOUCAULT, Michel "La tecnología política de los individuos". Veredas 5,9 (2004): 213-226. GENERALITAT DE CATALUÑA (2017). “Impuesto a las bebidas azucaradas”. HUMPHREY, Albert (2005), “SWOT Analysis for Management Consulting”. SRI Alumni Newsletter. LAVAL, Christian y DARDOT, Pierre (2013). La nueva razón del mundo. Ensayo sobre la sociedad neoliberal. Barcelona: Gedisa. LÉVI-STRAUSS, Claude (1969). Las estructuras elementales del parentesco. Traducción de Marie Thèrese Cevasco. Barcelona: Paidós. MACEY, David (1995). Las vidas de Michel Foucault. Traducción de Carmen Martínez. Madrid: Cátedra. MACHEREY, Pierre (2011). De Canguilhem a Foucault: la fuerza de las normas. Traducción de Horacio Pons. Buenos Aires : Amorrortu. OMS (2015). Plan de acción mundial sobre la resistencia a los antimicrobianos. PÚBLICO (2017). “¿Por qué los hombres matan a las mujeres? O cómo desvirtuar la violencia machista”. SANDÍN, Bonifacio. “DSM-5: ¿cambio de paradigma en la clasificación de los Trastornos mentales?”. Revista de Psicopatología y Psicología Clínica 18, 3 (2013):255-286. KAMCHATKA 10 (DICIEMBRE 2017): 13-38 38.	8,521
https://openalex.org/W2543931167_1	Spanish-Science-Pile	Open Science	Various open science	2,016	John James Park, Los Dogmas de la Constitución, edición y estudio preliminar de Joaquín Varela Suanzes-Carpegna, traducción del inglés y epílogo de Ignacio Fernández Sarasola, Tecnos, Madrid, 2015	None	Unknown	Unknown	3,823	6,881	John James Park, Los Dogmas de la Constitución, edición y estudio preliminar de Joaquín Varela Suanzes-Carpegna, traducción del inglés y epílogo de Ignacio Fernández Sarasola, Tecnos, Madrid, 2015 (J. J. Park, O el golpe definitivo a los teóricos de la balanced constitution). doi: 10.18543/ed-64(1)-2016pp413-421 I En el año 1605, Miguel de Cervantes Saavedra, de quien este año 2016 se conmemora el cuarto centenario de su fallecimiento, publicaba la primera parte de su novela El ingenioso hidalgo don Quijote de la Mancha. Es bien conocido el argumento de la obra, por lo que no es el momento ni el lugar de entrar en el mismo ni en profundizar en las posibles intenciones del autor al darla a luz, que el hoy injustamente olvidado Ramiro de Maeztu desgranó en su magistral ensayo incluido en el libro Don Quijote, don Juan y la Celestina; lo importante es que en la novela se mostraba cómo un hidalgo manchego de finales del siglo xvi se empeñaba en ajustar su conducta a un lenguaje y unos códigos caballerescos propios de la Alta Edad Media que, aun cuando siglos atrás constituyeron principios y normas admitidos por la caballería medieval, habían perdido su vigencia ya a mediados del siglo xv, no digamos ya en los umbrales del siglo xvii. El efecto de la obra fue fulminante. Si aún en pleno siglo xvi, cuando ya se había superado con creces la época del ideal caballeresco, se publicaron todavía algunas novelas que ensalzaban los usos de la caballería –buena prueba de ello son tanto la reelaboración del Amadís de Gaula debida a Garci Rodríguez de Montalvo como las secuelas a que dicha obra dio lugar, entre las que destaca sobremanera Las sergas de Esplandián- el ataque efectuado en la obra cervantina ocasionó la total desaparición de las novelas de caballerías, incapaces de sobreponerse no ya a la realidad, sino al mortal golpe que Cervantes les había asestado. Puede decirse que algo similar acaeció en el campo del análisis constitucional en el Reino Unido al finalizar el primer tercio del siglo xix. En pleno desarrollo industrial y con un imperio que se extendía prácticamente por todo el orbe, aún podían encontrarse tratadistas que continuaban analizando el sistema político inglés bajo los esquemas de la balanced constitution teorizados por John Locke y sancionados jurídicamente Estudios de Deusto 1 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 413 John James Park, Los Dogmas de la ConstituciónTecnos en el Bill of Rights de 1689. No obstante, a lo largo del siglo y medio transcurrido entre 1689 y 1832, las mutaciones constitucionales que tuvieron lugar en el Reino Unido, consistentes en el desapoderamiento de facto del monarca en favor de un gabinete responsable ante la Cámara de los Comunes (mutaciones que tuvieron lugar al margen del derecho escrito) tuvieron dos consecuencias fundamentales. La primera, en el orden político, al producirse un divorcio entre la realidad constitucional y el derecho escrito que supuso en el ámbito de la teoría política el paso de una monarquía constitucional a una monarquía parlamentaria. El segundo, y buena parte consecuencia del anterior, es la división de los publicistas británicos entre quienes aún a finales del siglo xviii y comienzos del siglo xix persistían en analizar la constitución inglesa sobre la base del derecho escrito orillando las mutaciones producidasy quienes en sus reflexiones se hacían eco de esos cambios políticos que de facto alteraron la constitución inglesa; en otras palabras, entre quienes veían a Gran Bretaña como una monarquía constitucional y quienes la contemplaban como una monarquía parlamentaria. No obstante, en el año 1832 se va a publicar una obra que, como el Quijote para los libros de caballerías, va a tener un efecto fulminante para los teóricos de la balanced constitution, con la única diferencia que ni el autor ni la obra iban a gozar de la misma fama y reconocimiento que el escritor alcalaíno; nos estamos refiriendo a The dogmas of the Constitution, de John James Park. Esta obra, descono- cida fuera del ámbito inglés y no suficientemente valorada en su país, ha tenido una importancia fundamental, al liquidar definitivamente la visión del sistema político inglés efectuada bajo los esquemas de una monarquía constitucional caracterizada por el equilibrio de Rey, Cámara de los Comunes y Cámara de los Lores. Bien es cierto que existían ilustres precedentes que ya se hacían eco de algunas de las instituciones del naciente parlamentarismo, pero el efecto que la obra de Park tuvo para consolidar los análisis efectuados bajo la óptica de la distinción entre realidad constitucional y derecho escrito (y que culminarán, de alguna forma, en Walter Bagehot y su The English Constitution) fue decisiva. II Si hemos de buscar un término apropiado para describir la década de los años treinta del siglo xix desde la óptica del constitucionalismo, sin duda algunala palabra más apropiada es la de cambio. Basta para ello echar un vistazo a la situación de las principales naciones del orbe. España aún continuaba, es cierto, regida por la aplastante bota del rey Fernando VII (por utilizar una feliz expresión del profesor Sosa Wagner), pero un análisis más detenido permitía otear ya un atisbo de esperanza para quienes deseaban un régimen constitucional, pues de forma silenciosa algunos próceres del liberalismo más templado se habían ido situando en las cercanías del trono, tomando posiciones para el momento en el que el monarca pasase a mejor vida; y, en efecto, así ocurrió Estudios de Deusto 414 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 2 John James Park, Los Dogmas de la ConstituciónTecnos cuando apenas seis meses después del óbito del rey felón, el gobierno presidido por Francisco Martínez de la Rosa aprobara el Estatuto Real. En Francia el cambio se produjo por vía revolucionaria, mediante el derrocamiento del rey Carlos X y su sustitución por un integrante de la rama de los Orleáns, Luis Felipe, con una liberalización del sistema de la Carta otorgada por Luis XVIII en 1814, que formalmente se mantenía. Bélgica se rebelaba proclamando su independencia y aprobando su Constitución ese mismo año 1830. Incluso en la otra orilla del Atlántico, en las antiguas colonias británicas mutadas en Estados Unidos, cuando el primer mandato del presidente Andrew Jackson tocaba a su fin y afrontaba en ese año 1832 unos comicios presidenciales que le otorgarían un segundo período al frente de la Jefatura del Estado, se produjo un cambio notable: la transición del sistema original en el que predominaba un cierto elitismo republicano hacia lo que se ha calificado por la historiografía como «democraciajacksoniana»; y es que fue precisamente bajo los ocho años en los queOld Hickory, el héroe de la batalla de Nueva Orleáns, ocupó la presidencia de los Estados Unidos cuando surgen en dicha nación los partidos políticos en el moderno sentido del término y el momento en el que asienta sus bases la moderna democracia estadounidense. Pues bien, esos aires de renovación política (en ocasiones a través de la vía revolucionaria, en otras por vía reformista) también afectaron y no poco al Reino Unido de Gran Bretaña, que precisamente en ese año 1832, es decir, el mismo en el que John James Park publica su obra sobre la Constitución inglesa, afronta un cambio legal decisivo para asentar a nivel de derecho positivo el parlamentarismo que ya de facto caracterizaba su sistema. III Gran Bretaña afronta en 1832 un momento determinante para asentar el parlamentarismo a través de una reforma legal de hondo calado que tiene precisamente eco en la obra de Park, por lo que conviene hacer una breve recapitulación de lo acaecido durante los ciento cincuenta años anteriores, es decir, desde la Glorious Revolution de 1688. No importa, a estos efectos, determinar si se trató de una revolución conservadora y pacífica, como nos indica la historiografía tradicional heredera de Macaulay o si, por el contrario, fue una revolución moderna y violenta, como defiende Steve Pincus en su reciente y detallado estudio analítico del evento, que titula significativamente 1600: La primera revolución moderna. Lo relevante es que tanto a nivel teórico (fundamentalmente gracias a John Locke) como a nivel jurídico se articula una monarquía constitucional garantizada por el equilibrio Rey-Comunes-Lores pero que, a diferencia de lo que ocurrirá en otros países, no culmina en un texto constitucional escrito. Ahora bien, tras la muerte de la reina Ana en 1714 y la subida al trono de Jorge I, príncipe elector de Hannover y primer monarca de tal dinastía, va a tener lugar una silenciosa y progresiva mutación constitucional que opera al margen del de- Estudios de Deusto 3 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 415 John James Park, Los Dogmas de la ConstituciónTecnos recho escrito y a tenor de la cual se va a desplazar la dirección política del Estado de las manos del rey a las de un gabinete responsable ante el Parlamento. Durante el largo mandato de sir Robert Walpole van a tener lugar dos circunstancias determinantes para la transformación progresiva de la monarquía constitucional en parlamentaria. Una de ellas la aparición en el Gabinete de la figura de una figura, el primer ministro, que va a gozar de preferencia sobre el resto, aspecto ésteen el que tuvo no poco que ver la recia personalidad de Walpole. La segunda, el inicio de la preponderancia de la Cámara de los Comunes sobre la de los Lores o, lo que es lo mismo, del elemento popular sobre el aristocrático; así, el 1 de febrero de 1739 Sir Robert Walpole reconocía precisamente ante la Cámara de los Comunes que: «This House and Parliamentishis Majesty´sgreatest, safest, and bestcouncil. A los cambios políticos se unieron unas alteraciones sociales que influyeron igualmente en el plano institucional. La revolución industrial que tuvo lugar a finales del siglo xviii y primer tercio del siglo xix, mutó notablemente la faz de la nación inglesaen todos los ámbitos. El auge de nuevos establecimientos fabriles e industriales implicó la aparición de un nuevo paisaje urbano, de una nueva clase trabajadora y, sobre todo, grandes bolsas de proletariado industrial. Es la realidad social tan majestuosamente descrita por Charles Dickens (impagable la forma de reflejaresa nueva realidad industrial coetánea que personifica en la Coketown desu novela Tiempos difíciles, o su peculiar visión de las actividades comerciales y educativas que expone en Nicholas Nickleby, magníficas obrasque no son quizá las Estudios de Deusto 416 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 4 John James Park, Los Dogmas de la ConstituciónTecnos más conocidas o populares del autor) y que Carlos Marx tomaría ulteriormente para la elaboración de sus teorías económicas anticapitalistas. Pero sobre todo, esa nueva realidad industrial provocó una serie de migraciones interiores que determinarían los inicios de la reforma parlamentaria. La Cámara de los Comunes, que se había erigido en el nervio del sistema político, era elegida conforme a un sistema de representación anclado en la realidad preindustrial, de tal manera que ciudades sin apenas entidad continuaban enviando numerosos representantes a la misma mientras otras que habían incrementado notablemente su población al haberse constituido en importantes centros industriales, o bien carecían de representación o ésta no era en modo alguno proporcional a la importancia adquirida por tales núcleos urbanos. Esa disfunción es la que tratará de superarse con la Reform Act de 1832. La peculiar situación inglesa anteriormente descrita, es decir, la carencia de un texto constitucional escrito y el divorcio entre la realidad y el derecho escrito, es la que va a propiciar una división entre los autores que se mantenían anclados en las visiones tradicionales de la balanced constitution, y continuaban viendo en Inglaterra un ejemplo de monarquía constitucional y equilibrio de poderes caracterizado por la igualdad entre monarca, lores y comunes (el caso paradigmático sería el de sir William Blackstone y sus CommentariesontheLaws of England, o ya más tarde el de William Paley), y por otra parte quienes no cerraban los ojos a la realidad cotidiana e incidían en el predominio del elemento parlamentario y en la institución del gabinete (caso de Edmond Burke y Jeremy Bentham). Es curiosamente en este momento de confluencia entre los debates políticos sobre la reforma parlamentaria y la división doctrinal que acabamos de mencionar, cuando un prestigioso abogado y profesor del King’s College va a pronunciar una serie de conferencias en el seno de un curso de Derecho, cuatro de las cuales va a recoger en forma de libro. El autor en cuestión era John James Park y el libro no es otro que The dogmas of the Constitution. IV Los dogmas de la Constitución es una obra que fue publicada en Españapor vez primera en el año 1998 en la editorial Itsmo, con un amplio estudio preliminar de Joaquín Varela Suanzes-Carpegna, y pese a tratarse de la primera traducción en lengua no inglesa de la obra de Park, desgraciadamente pasó casi desapercibida, encontrándose además agotada desde hace años. Por fortuna, dicho libro acaba de ser reeditado por la editorial Tecnos en su prestigiosa colección Clásicos del Pensamiento Político, y a la detallada introducción del profesor Varela (actualizada bibliográficamente) se añade ahora a modo de epílogo un interesantísimo estudio de Ignacio Fernández Sarasola que profundiza en esa dicotomía entre teoría y práctica constitucional mediante una exposición de la tesis de los principales analistas políticos británicos desde el siglo xviii hasta bien entrado el siglo xx. Joaquín Varela, como indica en las Estudios de Deusto 5 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 417 John James Park, Los Dogmas de la ConstituciónTecnos primeras páginas de su estudio, ha tenido que efectuar una labor «casi detectivesca» para ofrecer al lector un esbozo biográfico de Park, labor muy meritoria si tenemos en cuenta la casi ausencia de fuentes bibliográficas sobre el personaje. Gracias a tal esfuerzo podemos conocer,aunque seaen sus rasgos básicos, la vida y obra de Park, autor a quien el profesor Varela sitúa magistralmente en las coordenadas político-culturales del momento y al que inserta como una pieza más en el amplio mosaico de autores que dedicaron sus esfuerzos a desentrañar las esencias del sistema político inglés. A nivel estrictamente personal quisiera destacar dos aspectos que considero fundamentales para entender en su totalidad al autor y su obra. El primero, la en ocasiones machacona insistencia de Park explicitando esa disfunción o diferencia entre la realidad política inglesa y su derecho escrito, dado que no desaprovecha ocasión para incidir en esa dicotomía tan esencial a la historia reciente de Gran Bretaña. La segunda circunstancia, que a quien suscribe es particularmente cara, es la condición de jurista práctico de Park, pues el mismo unía a su categoría de profesor universitario la de abogado en ejercicio, y creo no es una circunstancia nada baladí que en la portada de su obra el autor anteponga significativamente su condición de barrister at law a la de profesor universitario. En este sentido, en su estudio preliminar Joaquín Varela señala esta condición y hace referencia a la distinción entre solicitor y barrister existente en la abogacía inglesa, dualidad esencial al sistema británico y desconocida en otros países. Esto últi- mo me parece determinante, pues consideroampliamente acreditado que la visión de un profesor universitario difiere notablementede la que tiene un abogado en ejercicio, de igual manera que un letrado no aborda el asunto de la misma forma ni con la misma perspectiva de un juez. Conviene recordar que el libro comentado se limita a recoger cuatro de Estudios de Deusto 418 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 6 John James Park, Los Dogmas de la ConstituciónTecnos las lecciones pronunciadas por el autor en el King’s College, a las que añade un prefacio y dos apéndices. Ya desde las páginas iniciales antepuestas al grueso de la obra, Park enuncia la que va ser la tesis central que aparece una y otra vez a lo largo de la misma, es decir, la distinción entre realidad constitucional y derecho escrito, tesis que pretende demostrar, además,ajeno a toda influencia política, es decir, considerándose a sí mismo distantetanto de la ideología whig como de la tory, aspecto éste que igualmente aparece disperso a lo largo de las páginas del libro y que supone, en cierto modo, una excusatio nonpetita. Esa idea o eje central sobre el que pivota toda la obra, creo la resumen perfectamente dos breves frases insertas en la lección décima (la segunda de las recogidas en la obra) donde se plasma claramente el influjo del positivismo en el autor: «Caballeros, hay dos tipos de teoría, de igual manera que existen dos tipos de filosofía. La teoría o filosofía que prescribe leyes y principios de su propia invención, y la teoría o filosofía que comprueba las leyes y principios mediante el aprendizaje de la observación.» Ya está, pues, consagrada la dualidad teoría-práctica, que aplicada al ámbito jurídico supone distinguir entre principios consagrados en las leyes por un lado y realidad constitucional inglesa por el otro, es decir, lo que apenas ochenta años más tarde José Ortega y Gasset calificaría en su ensayo Vieja y nueva política, aplicado a nuestro país, como España oficial y España vital. Park dejaba clara esta obsesiva idea suya en los párrafos iniciales de la primera de las lecciones, aunque con una evidente exageración al presentarse poco menos que como el pionero en la defensa de dicha tesis: «La Constitución propositiva o teórica de Gran Bretaña (si ha existido en un estado puro, lo cual resulta dudoso), ha dejado de tener existencia desde hace siglo y medio, y ha sido sustituida por una maquinaria totalmente diferente. Pero este hecho no ha sido jamás reconocido o registrado públicamente. La Constitución sustituta no ha sido en ningún momento reducida formalmente a una proposición […] De esta manera, cuando trato del fundamento de la Constitución, mi propósito es analizarla en atención a su realidad; a saber, que durante al menos los últimos ciento cincuenta años ha habido dos Constituciones concurrentes pero esencialmente distintas que han coexistido, aunque sin reconocimiento expreso de tal circunstancia; una en sustancia, la otra tan sólo en la forma.» He aquí resumido en breves palabras toda la intención final del libro: dejar constancia de la distinción entre norma y realidad, entre derecho escrito y derecho vivo. Una transformación que se había producido de forma gradual y silente, como indica en este lúcido párrafo incluido en la decimotercera lección (la cuarta y última de las que recoge en la obra): «Toda la Constitución del país se ha transformado por completo en los últimos ciento cincuenta años. Pero, al haberse observado rígidamente sus formas en muchos aspectos, esta revolución se ha producido de forma silenciosa e imperceptible ante los ojos de los cronistas y tratadistas de la Constitución que se dejan llevar por el sonido de las palabras sin atender a la eficacia de las cosas. Les harán creer que la Constitución aún exis- Estudios de Deusto 7 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. Pese a la decisiva influencia en la doctrina posterior, el éxito no acompañó a la obra de Park (quien fallecería justo un año después, en 1833, a la temprana edad de treinta y ocho años), que se sumió en el silencio de la historia incluso en su patria natal. Pero sus tesis acabaron imponiéndose y de ahí la importancia de esta obra que tuvo el mérito indiscutible, como indica el profesor Fernández Sarasola, de «poner de manifiesto con tanta evidencia la discrepancia existente entre el statutelaw y la realidad política de Inglaterra; entre los planteamientos teóricos de los tratadistas más exitosos (de Montesquieu a Paley) y el verdadero funcionamiento del sistema de gobierno de Albión.» Y el golpe que asestó Park a las anticuadas teorías de la balanced constitution fue tan mortal como el asestado dos siglos y cuarto atrás por Cervan- tes a las no menos anacrónicas novelas de caballerías. V Pese a que toda obra, ya sea literaria o política, indudablemente está ligada a las coordenadas socio-temporales en que se ha elaborado, no es menos cierto que existen reflexiones, frases, párrafos o ideas que trascienden a las mismas y pueden aplicarse a cualquier momento o lugar. De hecho, en esta obra existe un párrafo que cobra rabiosa actualidad, y que hace referencia al distanciamiento entre la política y la ciudadanía así como las consecuencias que de ello pueden derivarse. Juzgue el lector si estas palabras, contenidas en la primera de las lecciones, no podrían perfectamente haber sido escritas en 2016 y no en el año 1832: «No puedo sino pensar que uno de los grandes errores que se han cometido en este país es que los hombres de Estado, personas conocedoras del ejercicio práctico del gobierno, no han considerado cuán prudente es poner a la comunidad al corriente de los principios reales y del mecanismo del gobierno en la práctica, sino que han mantenido estos principios en sus propias entrañas a modo de una especie de arcana o conocimiento oculto (como era común en la medicina de hace cincuenta años), como si se tratase de algo que no fuera conveniente que el pueblo conociese. La consecuencia ha sido una creciente falta de comunión de sentimientos y entendimiento entre los hombres de Estado y el pueblo, cuando es posible que, de haber sido adecuadamente instruido o iniciado el público, ambos Estudios de Deusto 420 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 8 John James Park, Los Dogmas de la ConstituciónTecnos pudieran haber caminado juntos en muchas cuestiones. Además, el pueblo tendrá nociones, tendrá ideas sobre todas las cuestiones y se guiará por ellas. Me estoy refiriendo a la ya mítica Yes, minister en cuyo capítulo inicial, significativamente titulado Open government, se denunciaba precisamente que dicho lema electoral no servía para otra cosa que para disfrazar la misma realidad denunciada por John James Park en el King´s College, lo que se hacía de forma elegante y divertidísima través de un diálogo entre el Secretario Privado del ficticio Ministerio de Asuntos Administrativos y el Secretario Permanente del Primer Ministro inglés. Así, mientras aquél preguntaba: «What´s wrong with open government. I mean. Why shouldn´t the public know more about what´s going on?», el segundo respondía: «Are you serious? […] It´s a contradiction in terms. You can be open or you can be government», precisando más tarde la razón de esa dicotomía: «If people don´t know what you are doing, they don´t know what you are doing wrong». Jorge Pérez Alonso Universidad de Oviedo Estudios de Deusto 9 ISSN 0423-4847 • ISSN-e 2386-9062, Vol. 64/1, Enero-Junio 2016, págs. 413-421 doi: 10.18543/ed-64(1)-2016pp413-421 • http://www.revista-estudios.deusto.es/ • © Universidad de Deusto 421.	48,709
https://openalex.org/W2983871045	OpenAlex	Open Science	CC-By	2,019	NEW INSIGHTS FOR STATISTICAL ANALYSIS OF BLOOD PRESSURE RESPONSE TO RESISTANCE TRAINING IN ELDERLY HYPERTENSIVE WOMEN	Dahan da Cunha Nascimento	English	Spoken	7,189	14,235	ABSTRACT The main goal was to present statistical procedures for a better data interpretation of responsiveness, explain how to deal with RTM effect, and describe how to determine clinically important changes in BP from significant real difference (SRD). Twenty-seven hypertensive elderly women were included, and RT consisted of a periodized linear model. The RT lasted 10 weeks, with two sessions performed per week. Responders were classified on the basis of SBP differences between time- points T1 (first 3 weeks) and T4 (weeks 9–10). Statistical analyses were performed using One-Way Repeated Measures ANOVA, an analysis of covariance (ANCOVA), the linear mixed model (LMM) was used in the present study, and SRD was also calculated. In conclusion, when one-way repeated measure ANOVA was conducted to determine whether there was a statistically significant difference in SBP levels over the course of 10-week RT, results showed a non-significant reduction of -2.24 mmHg, while classifying subjects by responsiveness provides a different perspective of the results. Furthermore, initial SBP was the more powerful predictor of post-exercise SBP response, as analyzed by the regression to the mean effect. Finally, the reductions of -2.24 mmHg was not statistically significant nor clinically meaningful, but fell within the measurement error of the SBP measurements. ords: Responsiveness, Regression to the mean, Standard error of measurement, Clinical difference, Blood pressure. Keywords: Responsiveness, Regression to the mean, Standard error of measurement, Clinical differenc NEW INSIGHTS FOR STATISTICAL ANALYSIS OF BLOOD PRESSURE RESPONSE TO RESISTANCE TRAINING IN ELDERLY HYPERTENSIVE WOMEN NOVAS PERPECTIVAS PARA A ANÁLISE ESTATÍSTICA DA RESPOSTA DA PRESSÃO ARTERIAL AO TREINAMENTO RESISTIDO EM MULHERES IDOSAS HIPERTENSAS Dahan da Cunha Nascimento1,4, Cristiane Rocha Silva1, Denis Cesar Leite Vieira2,4, Brad Jon Schoenfeld3 and Jonato Prestes1 1Universidade Católica de Brasília, Brasília-DF, Brasil. 2Universidade de Brasília, Brasília-DF, Brasil. 3Cuny Lehman College, Bronx-NY, United States of America. 4Centro Universitário do Distrito Federal, Brasília-DF, Brasil. 1Universidade Católica de Brasília, Brasília-DF, Brasil. 2Universidade de Brasília, Brasília-DF, Brasil. 3Cuny Lehman College, Bronx-NY, United States of America. 4Centro Universitário do Distrito Federal, Brasília-DF, Brasil. RESUMO O objetivo principal do estudo foi apresentar procedimentos estatísticos para uma melhor interpretação dos dados sobre a responsividade, explicar como lidar com o efeito da regressão a média (RM) e descrever como determinar alterações clinicamente importantes na pressão arterial (PA) pelo cálculo da diferença clínica (DC). Vinte e sete mulheres idosas hipertensas foram incluídas e o treinamento resistido (TR) consistiu em um modelo linear periodizado. O TR durou 10 semanas, com duas sessões realizadas por semana. Os responsivos foram classificados com base nas diferenças da pressão arterial sistólica (PAS) entre os momentos T1 (primeiras 3 semanas) e T4 (semanas 9–10). As análises estatísticas no presente estudo foram realizadas utilizando a ANOVA de medidas repetidas, análise de covariância (ANCOVA) e modelo linear misto (MLM). Conclui-se que quando uma ANOVA de medidas repetidas é aplicada, os resultados mostram uma redução não significativa de -2,24 mmHg, mas a classificação dos participantes por responsividade fornece uma interpretação diferente dos resultados. Além disso, a PAS inicial foi o preditor mais potente da resposta pós-exercício da PAS, conforme analisado pela RM. Finalmente, as reduções de -2,24 mmHg não foram estatisticamente significativas e nem clinicamente importantes, mas caíram dentro do erro de medida. p Palavras-chave: Responsividade, Regressão a média, Erro padrão de medida, Diferença clínica, Pressão arterial. DOI: 10.4025/jphyseduc.v30i1.3025 DOI: 10.4025/jphyseduc.v30i1.3025 DOI: 10.4025/jphyseduc.v30i1.3025 Artigo Original Introduction Exercise is beneficial for the control and prevention of hypertension in the elderly. In addition, it has been shown that traditional resistance training (RT) combined with aerobic training are effective in lowering blood pressure (BP) in elderly hypertensive women after 16 weeks1. Moreover, research indicates these benefits can be maintained for 4 and 14 weeks, respectively, following training cessation2,3. Those benefits promoted by the regular practice of RT might be explained by the changes on neural mechanisms, suh as lower renal and muscle sympathetic nerve activity, lower norepinephrine spill-over, increament on heart rate J. Phys. Educ. v. 30, e3025, 2019. Page 2 of 11 Page 2 of 11 Page 2 of 11 Nascimento et al. Nascimento et al. variability, and enhanced parasympathetic modulation that lower resting heart rate and blood pressure4. Despite the reported beneficial effects of exercise in lowering BP levels, this variable naturally fluctuates based on factors such as emotional status, stress, antihypertensive medications, inaccuracies in the measurement process, and illness5. Thus, the greater BP variability during the study intervention, the greater the error measurement for individual BP scores. When taking this information into account, the apparently simple question: “Does experimental group BP reductions provided by interventions represent a statistical significant difference, a real difference, or a measurement error?” becomes rather complex. The effects of exercise on blood pressure are normally represented by measures of central tendency (e.g. mean) when groups and time points are compared. This common practice is based on the premise that subjects will tend to present a similar response. However, in actuality subjects show a wide range of responses to an exercise intervention (a.k.a “responsiveness”)6. Thus, analyzing data using group or time points means might induce Type III7,8. For the Type III error, the wrong question for the right answer is made. Essentially the Type III error, occurs when the statistician fails to do the best job and has not taken enough time (e.g. use of adequate statistical analyses) to question the research experiment8. For the wrong question and misleading interpretation of a null hypothesis correctly rejected, researchers must be aware that BP reductions will occur for a small group of subjects. In addition, reductions in systolic blood pressure (SBP) will occur for subjects who show exceptionally large responses to training (a.k.a. “high responders”) when compared with individuals who show small responses to training (a.k.a. “low responders”). Introduction In this sense, looking at alternative statistical measures than just the group means and variability of data around the mean (e.g. standard deviation) might provide important insights as to inter- individual variation in training response9-14. Another concern involving blood pressure response to exercise, is the regression to the mean (RTM). Variables as BP, that is associated with large within-subject variability over time, create the potential for a RTM effect15. A higher BP in the first measurement (e.g., at first week), will be likely be less extreme when measured again (e.g., at 10 week). Thus, classifying subjects as high and low responders to exercise will, in the great majority of cases, be RTM and random variation5,15. These findings emphasize the importance of including analysis of covariance (ANCOVA) into study design to deal with RTM as proposed by Barnett et al.16. In addition, it is important to statistically determine if changes in SBP are real or influenced by error measurements6. The desired accuracy of BP error measurement in a study can be controlled by a complementary statistical analysis. Therefore, standard error of measurement (SEM)17, which controls the aggregate of factors that collectively affects the true value of the measurement, can be measured and the minimal important change or smallest real difference (SRD) can be calculated. The SRD determines what should be clinically important from what is known as the error of measurement18. The application of these complementary statistical analyses to researchers in the field of BP and exercise interventions is relevant. Thus, the aim of the current study was to present statistical procedures for a better data interpretation of responsiveness, explain how to deal with RTM effect, and to describe how to determine clinically important changes in BP from SRD. Methods To best illustrate concepts, we present real data from an experimental 10-week RT program, and the effect on SBP in hypertensive elderly women. This study was approved by J. Phys. Educ. v. 30, e3025, 2019. New insights for statistical analysis of blood pressure response Page 3 of 11 Page 3 of 11 the Institutional Research Ethic Committee of Catholic University of Brasília (UCB) (protocol 45648115.8.0000.5650/2016). Each subject was fully informed about the risks and benefits associated with participation in the present study and gave their written informed consent to participate. Twenty-seven (n = 27) hypertensive elderly women were included in this prospective exploratory study designed as a “pre-post”-intervention. All participants were recruited on a voluntary basis from the local community through posters and lectures about the study. Subject’s characteristics are presented in Table 1. According to the American College of Sports Medicine, subjects were considered untrained because they had no previous experience with resistance training (RT)19. Inclusion criteria for hypertensive subjects were as follows: women age ≥ 60 years and subjects who self-reported hypertension and had SBP and DBP below the threshold of hypertension stage 1 but were using antihypertensive medications20-22. Subjects were excluded if they had a history of heart failure, valvular or congenital disease, pacemaker implantation, osteo-articular disorders, if they were smokers, or were consuming alcohol. Table 1. Subjects’ characteristics* Characteristics High (N = 7) Low (N = 20) p value SBP, mmHg 126.04±11.04 (115.83-136.26) 118.08±9.09 (113.83-122.34) 0.12 DBP, mmHg 72.95±6.47 (66.96-78.94) 69.02±6.54 (65.96-72.08) 0.18 Heart rate, bpm 72.54±15.61 (58.10-86.98) 72.02±8.57 (68.01-76.04) 0.91 Age, years 69.14±6.79 (62.86-75.42) 68.01±5.23 (65.65-70.54) 0.67 Height, cm 1.52±0.03 (1.49-1.54) 1.56±0.06 (1.53-1.59) 0.09 Weight, kg 57.00±5.64 (51.78-62.22) 71.22±9.32 (68.85-75.58) 0.001 BMI, kg/m² 24.64±2.49 (22.33-26.94) 29.01±3.30 (27.55-30.65) 0.003 Leg press 10 RM, kg 33.14±5.08 (28.44-37.84) 38.54±6.84 (35.34-41.74) 0.06 Chest press 10 RM, kg 21.84±3.96 (18.17-25.51) 22.49±4.58 (20.34-24.63) 0.74 Leg extension 10 RM, kg 40.27±11.85 (29.31-51.24) 39.34±10.75 (34.31-44.38) 0.84 Low row 10 RM, kg 33.57±3.77 (30.07-37.06) 34.75±5.89 (31.99-37.50) 0.62 Leg curl 10 RM, kg 39.39±9.01 (31.05-47.73) 39.98±8.08 (36.20-43.76) 0.87 Note: SBP = systolic blood pressure, DBP = diastolic blood pressure, BMI = body mass index, RM = repetition maximum. * Values are expressed as means, SD (standard deviation), and 95% confidence interval (CI) Source: Authors Table 1. Methods Subjects’ characteristics* Characteristics High (N = 7) Low (N = 20) p value SBP, mmHg 126.04±11.04 (115.83-136.26) 118.08±9.09 (113.83-122.34) 0.12 DBP, mmHg 72.95±6.47 (66.96-78.94) 69.02±6.54 (65.96-72.08) 0.18 Heart rate, bpm 72.54±15.61 (58.10-86.98) 72.02±8.57 (68.01-76.04) 0.91 Age, years 69.14±6.79 (62.86-75.42) 68.01±5.23 (65.65-70.54) 0.67 Height, cm 1.52±0.03 (1.49-1.54) 1.56±0.06 (1.53-1.59) 0.09 Weight, kg 57.00±5.64 (51.78-62.22) 71.22±9.32 (68.85-75.58) 0.001 BMI, kg/m² 24.64±2.49 (22.33-26.94) 29.01±3.30 (27.55-30.65) 0.003 Leg press 10 RM, kg 33.14±5.08 (28.44-37.84) 38.54±6.84 (35.34-41.74) 0.06 Chest press 10 RM, kg 21.84±3.96 (18.17-25.51) 22.49±4.58 (20.34-24.63) 0.74 Leg extension 10 RM, kg 40.27±11.85 (29.31-51.24) 39.34±10.75 (34.31-44.38) 0.84 Low row 10 RM, kg 33.57±3.77 (30.07-37.06) 34.75±5.89 (31.99-37.50) 0.62 Leg curl 10 RM, kg 39.39±9.01 (31.05-47.73) 39.98±8.08 (36.20-43.76) 0.87 Note: SBP = systolic blood pressure, DBP = diastolic blood pressure, BMI = body mass index, RM = repetition maximum. * Values are expressed as means, SD (standard deviation), and 95% confidence interval (CI) Source: Authors Trial design g The strength of upper and low body was evaluated by ten-repetition maximum (10- RM) strength testing. After the 2-week familiarization period, subjects were tested for a 10- RM for the following exercises in this order: machine leg press, machine chest press, machine leg extension, machine low row, and machine leg curl (Righetto Fitness Equipment, Sao Paulo, Brazil) with 5 minutes rest between exercise tests. Subjects were advised to refrain from any exercise other than activities of daily living for at least 48 hours before 10-RM testing. In brief, subjects warmed-up on each exercise with 5-10 submaximal repetitions. Subjects performed 10 repetitions of increasing weight until reaching a valid 10-RM. Two minutes of rest was provided between attempts. All 10-RM tests were registered within two attempts. Two experienced RT professionals supervised the tests. Furthermore, subjects were evaluated by an experienced physiotherapist before the 10-RM testing and study participation. Previous studies from our research group demonstrated a high test and retest reliability for this type of test r > 0.971,11. All testing sessions were scheduled between the hours of 8:00 to 10:00 AM (morning class group) and 1:00 and 3:00 PM (afternoon class group). Before each training session, the correct use of hypertensive medications and the risks associated with non-use were reinforced23. J. Phys. Educ. v. 30, e3025, 2019. Hemodynamic measurements The SBP, diastolic BP (DBP) and heart rate were measured before each training session with an automatic oscillometric validated device (model Microlife BP 3BTO-A, Onbo Electronic, Widnau, Switzerland)26, consistent with the recommendations of the Guideline for the Prevention, Detection, Evaluation, and treatment of High Blood Pressure27,28. The measurements were performed after 10 minutes of seated rest in a quiet, temperature- controlled room, and cuff size was adapted to the arm circumference of each subject. All measurements of BP were taken between 8:00 and 10:00 a.m. (morning class group, 14 participants) and 1:00 and 3:00 p.m. (afternoon class group, 13 participants). Before measurements, subjects were advised to refrain from programmed exercise, and caffeine consumption. Page 4 of 11 Nascimento et al. The RT consisted of a periodized linear model. The exercises performed were: machine leg press, machine chest press, machine leg extension, machine low row, and machine leg curl. The number of repetitions were reduced (maintaining the minimal zone established for each cycle) as the intensity increased over the course of the program. The periodization scheme was adapted from our previous research described in detail elsewhere24. The RT lasted 10 weeks, with two RT sessions performed per week, with a minimum of 24 h between sessions. All sessions were supervised by experienced RT professionals with a supervision ratio of 1:1 (coach to participant ratio) to favor greater strength gains25 and for safety. Subjects were instructed to lift and lower loads at a constant velocity, taking ~2 seconds for the concentric (muscle shortening) and 2 seconds for the eccentric phase (muscle lengthening). In the first 3 weeks (T1) of the program, subjects performed three sets of 12–14 RM with a 60 s rest interval; from weeks 4–6 (T2), subjects performed three sets of 10–12 RM with a 80 s rest interval; from weeks 7–8 (T3), subjects performed three sets of 8–10 RM with a 100 s rest interval; and from weeks 9–10 (T4), subjects performed three sets of 6–8 RM with a 120 s rest interval. Loads were increased when subjects performed more than 3 repetitions in the third set beyond the prescribed RM zone. Participant sub-grouping Responders were classified on the basis of SBP differences between time-points T1 and T4. High responders were classified as a percent BP decline in the 25th percentile for the SBP. High responders were classified as systolic blood pressure decline ≥ -6.83 mmHg and low responders < -6.83 mmHg. Furthermore, responsiveness was considered a “rare event” and no clinical or physiological criteria have been established to define high responders and low responders for SBP29. Although, different terms are used by previous research as, “high responders”, “responders” or even “adverse responders” to regular physical exercise6,9,11. In the present study, the terms high and low responders were used. Use of smallest real difference First, standard error of measurement was calculated by the square root of the mean square error term from the One-Way Repeated Measures ANOVA. This information can be found inside the output presented by statistical program SPSS (Table 2). We next calculated the minimum difference (MD) to be considered real by the following formula17,18: √ (Equation 1). MD = SEM x 1.96 x √2, (Equation 1). MD = SEM x 1.96 x √2, The 1.96 value represents the z score associated with 95% IC. According to Weir17, the researcher might choose a different z score if a more liberal or conservative assessment is desired. The 80% confidence for BP is only suggested if 10-13 office measurements can be J. Phys. Educ. v. 30, e3025, 2019. New insights for statistical analysis of blood pressure response Page 5 of 11 Page 5 of 11 made5 with an accuracy about ± 5 mmHg of the ‘true’ blood pressure. Thus, a conservative assessment using 95% of confidence is recommended. g Using the data of Table 1, the SEM will be √33.28 = 5.76 mmHg. Thereafter, according to equation 1, MD = 5.76 x 1.96 x √2 = 15.91 mmHg. Using the data of Table 1, the SEM will be √33.28 = 5.76 mmHg. Thereafter, according to equation 1, MD = 5.76 x 1.96 x √2 = 15.91 mmHg. Thus, a decline of at least 15.91 mmHg on BP needs to occur to be confident at the 95% level that a change in SBP reflects a real change and not a difference that is within the expected measurement error of the SBP values. Table 2. One-way repeated measures ANOVA Note: SBP = systolic blood pressure; MSE = mean square error Source: Authors Source Type III Sum of Squares df Mean Square F Sig. SBP Sphericity Assumed 198.15 3 66.05 1.98 0.07 Greenhouse-Geisser 198.15 2.85 66.42 1.98 0.07 Huynh-Feldt 198.15 3.00 66.05 1.98 0.07 Lower-bound 198.15 1.00 198.15 1.98 0.07 Error (SBP) Sphericity Assumed 2596.57 78 33.28 (MSE) Greenhouse-Geisser 2596.57 74.28 34.99 Huynh-Feldt 2596.57 78.00 33.28 Lower-bound 2596.57 26.00 99.86 Table 2. One-way repeated measures ANOVA Table 2. One-way repeated measures ANOVA Page 6 of 11 Nascimento et al. Nascimento et al. Kenward–Roger degrees of freedom approximation) and the parameters information in Table 5. All analyses were conducted with SPSS version 18.0 (SPSS Inc., Chicago, USA). In addition, GraphPad Prism 6.0 software was also used for analysis (San Diego, California, USA). Statistical analysis Data were expressed as means and standard deviations. Shapiro-Wilk and Mauchly’s test were employed to check for normality and sphericity, respectively. Levene’s test was used to test homogeneity of variances. When the assumption of sphericity was not met, the significance of F-ratios was adjusted according to the Greenhouse-Geisser correction. Initially a repeated measures ANOVA was employed to verify the effects of training on hemodynamic changes30. This statistical analysis considered the data of all subjects. An analysis of covariance (ANCOVA) controlling for pre-training SBP to verify its effects on SPB response post-training was employed to analyze the RTM effect16,31. Levene’s test was also applied for testing homogeneity of variances. When differences were identified, the Bonferroni post-hoc test was applied. An alpha level of ≤ 0.05 was considered significant, and p values were two-tailed. Considering a partial eta squared of 0.35, effect size of 0.74, α error probability of 0.05, total sample size of 27 participants, number of groups (two groups), and covariates (one covariate), the power provided was 0.95% for this statistical analysis. The software GPower 3.1.6 was used to calculate the power. Considering that sphericity condition is very difficult to justify, and it is very unlikely that the measurements taken between time points T1 to T4 will have the same degree of correlation. An analysis that can represent the average response value of the response at any time point in term of covariates such as treatment group (responsiveness), 10-week RT program, pre-training SBP values, and also account successfully for the observed pattern of dependences in those measurements. The linear mixed model (LMM) also called mixed effect model (MEM), random-regression models, multilevel models, hierarchical linear models, and empirical Bayes models32 was used in the present study and can properly account for correlation between repeated measurements on the same subject and have greater flexibility to model time effects32,33. In addition, the LMM are expected to provide more accurate statistical inference than ANOVA, and have a power above 80%34. We controlled for the influence of covariates (responsiveness, class time, 10-week RT program, and pre-training SBP values), that might predict post-training SBP changes. For the sake of brevity, we presented the F tests from the LMM results (type III Wald F tests with J. Phys. Educ. v. 30, e3025, 2019. All data analyzed All data analyzed y A one-way repeated measure ANOVA was conducted to determine whether there was a statistically significant difference in SBP levels over the course of 10-week RT program. The RT intervention did not elicit statistically significant changes in SBP levels over time (F(3.00, 78.00) = 1.98, p = 0.071). Figure 1. Figure 1. Systolic blood pressue response Note: significantly different from low responders at the same time point (p < 0.05). Source: Authors Figure 1. Systolic blood pressue response g y p p Note: significantly different from low responders at the same time point (p < 0.05). Source: Authors J. Phys. Educ. v. 30, e3025, 2019. Smallest real difference ff After evaluating the SRD, only one participant from the high responders group presented a real change on SBP of -23.92 mmHg (Table 4). Other participants from the high responders group were within the error measurement (-9.66 ± 2.54 mmHg). Similarly, only one subject from the low responders group presented a real change in SBP of 24 mmHg (Table 4). Table 4. Mean change score for SBP values according to the SRD Systolic blood pressure n Mean change value (SD) High Responders NSRD 6 -9,66 ± 2.54 mmHg SRD 1 -23.92 mmHg Low Responders NSRD 19 -0,17 ± 4.83 mmHg SRD 1 24.83 mmHg Note: SRD = smallest real difference, NSRD = no smallest real difference, SD = standard deviation Source: Authors Table 4. Mean change score for SBP values according to the SRD Systolic blood pressure n Mean change value (SD) After LMM was applied, we found a significant effect of pre-training SBP values (F(1,76) = 58.41, p = 0.001) as predictive of post-training SBP changes. For other variables as class time (F(1,76) = 0.57, p = 0.450), responsiveness (F(1,76) = 0.24, p = 0.624), and 10- week RT program (with values, T2, T3, and T4), F(1,76) = 0.88, p = 0.350, no significant effects were observed. Table 5. 5. Parametes estimates of the model (LMM, or MEM) to estimate post-training SBP changes Table 5. Parametes estimates of the model (LMM, or MEM) to estimate post-tra changes Table 5. Parametes estimates of the model (LMM, or MEM) to estimate post-training SBP changes Parameters Estimate SE p 95% CI Class time -1,66 2,95 0,57 -7.77 – 4.44 10-week RT program -0,94 0,80 0,24 -2.56 – 0.67 Responsiveness -1,21 3,32 0,71 -8.09 – 5.66 Pre-training SBP 0,67 0,11 0,00 0.42 – 0.91 Note: LMM = linear mixed model, RT = resistance traininig, SBP = systolic blood pressure, SE = standard error, CI = confidence interval, MEM = mixed effect model Source: Authors Source: Authors Responsiveness analyzed After adjustment for pre-training SBP levels, there was a statistically significant difference in post-training SBP levels between groups (F(1.00, 24.00) = 13.17, p = 0.001) after ANCOVA analyses. Post-hoc analysis was performed with a Bonferroni adjustment. Post-training SBP levels were statistically lower in the high vs low responders group (mean difference of -11.44 (95% CI -17.95 to – 4.93) mmHg, p = 0.001). Table 3. ( ) g p ) Considering that differences between groups were verified for BMI, after adjustment for pre-training BMI values, there was no statistically significant difference in post-training SBP levels between groups (F(1.00, 24.00) = 0.37, p = 0.544). New insights for statistical analysis of blood pressure response Page 7 of 11 Page 7 of 11 Page 7 of 11 Table 3. Adjusted and unadjusted intervention means and variability for post-training systolic blood pressure values with pre-training systolic blood pressure as a covariate Unadjusted Adjusted N M SE M SE High responders 7 114.34 3.91 109.43 2.66 Low responders 20 119.15 2.31 120.87 1.52 Note: N = number of participants, M = mean, SE = standard error. * Significantly different from low responders (p ≤ 0.001) Source: Authors able 3. Adjusted and unadjusted intervention means and variability for post-training blood pressure values with pre-training systolic blood pressure as a covariate Unadjusted Adjusted Page 8 of 11 Nascimento et al. African and white Americans. Furthermore, Lawes et al.36 demonstrated that a 10 mmHg reduction in SBP at mean age at baseline of 63 years was associated with a risk reduction in stroke of 31%. Nevertheless, the reductions of -2.24 mmHg observed in Figure 1 were neither statistically significant nor clinically meaningful, but fell within the measurement error of the SBP measurements. African and white Americans. Furthermore, Lawes et al.36 demonstrated that a 10 mmHg reduction in SBP at mean age at baseline of 63 years was associated with a risk reduction in stroke of 31%. Nevertheless, the reductions of -2.24 mmHg observed in Figure 1 were neither statistically significant nor clinically meaningful, but fell within the measurement error of the SBP measurements. Alternatively, classifying subjects by responsiveness (Figure 1) provides a different perspective on results. Seven elderly hypertensive women were deemed high responders and 20 participants as low responders. Differences between groups were identified; SBP on time point T4 were significantly lower for high responders when compared with low responders. These differences were only observed because the variability of scores around the mean were considered37. However, differences between time points reveals nothing about the clinical significance of findings or error measurement. This can be confirmed by the study from Nascimento et al.2 who used an ANOVA of repeated measurements, and demonstrated a significant decline (-10 mmHg) on SBP at post-training, and detraining time points when compared with pre-training measurements in hypertensive elderly women. Nevertheless, important insights as to inter-individual variation in training response were missed by the authors. Furthermore, the signicant differecens between time points observed by Nascimento et al.2 might reveal little about clinical significance of the findings or error measurement. Thus, it is important to distinguish findings of statistical significance from what is clinically meaningful. Statistical significance is a statement about the likelihood of a result being due to random chance, but is not indicative of whether the finding is practically meaningful. On the other hand, the clinical meaningfulness of a finding has relevant implications for practical application regardless of whether the result is significant38. 9 Bouchard et al.9 classified responsiveness by the measurement error of the trait. The parameter that captures the totality of these sources of variance in a trait is known as the technical error (TE). Discussion In the above paragraphs, different statistical approaches has been discussed from an analytical, post hoc perspective, aiming at the identification of responders, how to verify the RTM effect, interpret the measurement error, and calculate the SRD effect. If the mean of 27 participants is used to compare reductions in SBP between time points, results would show a statistically non-significant reduction of -2.24 mmHg (Figure 1). Hardy et al.35 demonstrated that reductions of 2 mmHg in SBP were associated with reductions in the incidence of coronary artery disease, stroke, and heart failure events for J. Phys. Educ. v. 30, e3025, 2019. Page 8 of 11 Thus, they defined the adverse responders or low responders as subjects in which the response was 2 X TE in a direction indicating a worsening of the risk factor. For the SBP, exercise training-induced increaments of ≥ 10 mmHg. 13 The study from Moker et al.13 did not use any statistical approach to determine the totality of technical error. Although, responders decreased SBP by -11.5 mmHg, whereas no responders increased SBP by 7.9 mmHg after training. We can conclude if changes in SBP were real or influenced by error measurements. With regard to the aforementioned paragraphs, the results from the data used in the present study demonstrate that a change of at least 15.91 mmHg in SBP must occur to be confident at the 95% level that a change in SBP reflects a real change and not a difference that is within the expected measurement error of the SBP values. With this in mind, only one participant from the high responders group presented a real change on SBP of -23.92 mmHg (Table 4). Other participants from the high responders group were within the error measurement (-9.66 ± 2.54 mmHg). This implies that inter-individual variation response to exercise might cause a misleading interpretation. Similarly, only one subject from the low responders group presented a real change in SBP of 24 mmHg (Table 4). This increase represents an adverse response to regular exercise. Adverse response is defined as an exercise-induced change that worsens a risk beyond measurement error and expected day-to-day variation9,13. So, classifying individuals by responsiveness represents an important tool for identifying individuals with unwanted responses and provides the ability to personalize exercise prescription9. Furthermore, there is evidence that, independent of the exercise modality (resistance training, aerobic training, concurrent training)13,37, an adverse response is not a rare event, but in fact a common occurrence9. J. Phys. Educ. v. 30, e3025, 2019. New insights for statistical analysis of blood pressure response Page 9 of 11 Page 9 of 11 We also sought to assess the effect of RTM on SPB response. Analyses of covariance is one statistical method that have been proposed to estimate the RTM effect15. Using data from our study after adjustment for pre-training SBP levels, there was a statistically significant difference in post-intervention SBP levels between groups (F(1.00, 24.00) = 13.17, p = 0.001). In addition, after LMM, pre-training SBP was predictive of post-training SBP changes. Page 8 of 11 High and low responders changed SBP from T1 to T4 by -11.70 ± 5.86 mmHg (- 9.00%) and 1.07 ± 7.03 mmHg (1.10%), respectively. Thus, responsiveness is a common occurrence as presented in previous research11,12,29,39 or might be a RTM effect? The focus of the present study is to demonstrate that determining the existence of change in BP and exercise research is a complex undertaking. However, several considerations regarding the control of other variables that affect responsiveness as, mode of exercise, intensity, frequency, duration, genetic endowment, age, epigenetics, and baseline phenotype must be considered6,40. Yet, few researchers acknowledge the issue in their published studies, and attempts to quantify individual response are usually deficient41. Furthermore, the cut-off adopted in this study and previous research11,12,29,39 is another point of criticism and from the perspective of practical application a better criterion for the discrimination of responders and non-responders by the use of SRD is suggested, however this approach make the classification of subjects as responders more conservative40. In summary, in order to avoid misleading interpretation of results and obtain useful practical information, the complementary statistics presented herein as responsiveness classification, regression to the mean, standard error of measurement, and minimal difference are particularly suitable when hemodynamic parameters are analyzed. Conclusions When one-way repeated measure ANOVA was conducted to determine whether there was a statistically significant difference in SBP levels over the course of 10-week RT, results showed a non-significant reduction of -2.24 mmHg, while classifying subjects by responsiveness provides a different perspective of the results. Furthermore, initial SBP was the more powerful predictor of post-exercise SBP response, as analyzed by the regression to the mean effect. Finally, the reductions of -2.24 mmHg were not statistically significant nor clinically meaningful, but fell within the measurement error of the SBP measurements. This approach demonstrates the importance of incorporating SRD to determine what should be clinically important from what is known as the error of measurement. 1. Dos Santos ES, Asano RY, Filho IG, Lopes NL, Panelli P, Nascimento Dda C, et al. Acute and chronic cardiovascular response to 16 weeks of combined eccentric or traditional resistance and aerobic training in elderly hypertensive women: a randomized controlled trial. J Strength Cond Res 2014;28(11):3073-3084. Doi: 10.1519/JSC.0000000000000537 2. Nascimento Dda C, Tibana RA, Benik FM, Fontana KE, Ribeiro Neto F, Santana FS, et al. Sustained effect of resistance training on blood pressure and hand grip strength following a detraining period in elderly hypertensive women: a pilot study. Clin Interv Aging 2014;9:219-225. Doi: 10.2147/CIA.S56058. eCollection 2014. References Garcia P, Nascimento DD, Tibana RA, Barboza MM, Willardson JM, Prestes J. 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Doi: 10.1152/japplphysiol.00714.2014. 40. Hecksteden A, Kraushaar J, Scharhag-Rosenberger F, Theisen D, Senn S, Meyer T. Individual response to exercise training - a statistical perspective. J Appl Physiol 2015;118(12):1450-1459. Doi: 10.1152/japplphysiol.00714.2014. 41. Hopkins WG. Individual responses made easy. J Appl Physiol 2015;118 (12):1444-1446. Doi: 10.1152/japplphysiol.00098.2015. Acknowledgments: The first author wants to dedicate this paper to his family (Rita de Cassia and Nicolas Cunha). The author Dahan da Cunha Nascimento also acknowledge the financial support from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). ORCID dos autores: Dahan da Cunha Nascimento: 0000-0002-6580-9404 Cristiane Rocha Silva: 0000-0003-3558-8804 Denis Cesar Leite Vieira: 0000-0002-0761-1846 Brad Jon Schoenfeld: 0000-0003-4979-5783 Jonato Prestes: 0000-0003-0399-8817 Received on Mar, 17, 2018. Reviewed on Sep, 22, 2018. Accepted on Oct, 20, 2018. Author address: Dahan da Cunha Nascimento. Programa de Pós-Graduação em Educação Física, Universidade Católica de Brasília - Q.S. 07, Lote 01, EPTC – Bloco G. Código Postal: 71966-700 – Distrito Federal, Brasília, Brazil. Telefone: 61 3356 9350 - Fax: + 21/55/61 3356 9350. E-mail: dahanc@hotmail.com J. Phys. Educ. v. 30, e3025, 2019.	47,671
https://openalex.org/W3172914220	OpenAlex	Open Science	CC-By	2,021	Effect of Different Management Techniques on Bird Assemblages on Rice Fields in the Republic of Korea	Green Choi	English	Spoken	7,221	13,482	Effect of Different Management Techniques on Bird Assemblages on Rice Fields in the Republic of Korea Green Choi Kyung Hee University Min Seock Do National Institute of Biological Resources Seok-Jun Son Korea Institute of Wildlife Ecology Hyung-Kyu Nam (  namhk2703@korea.kr ) National Institute of Biological Resources Research Article Keywords: bird assemblages, habitats, intensive practices, pesticide use, Korea Posted Date: May 27th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-551000/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Effect of Different Management Techniques on Bird Assemblages on Rice Fields in the Republic of Korea Green Choi Kyung Hee University Min Seock Do National Institute of Biological Resources Seok-Jun Son Korea Institute of Wildlife Ecology Hyung-Kyu Nam (  namhk2703@korea.kr ) National Institute of Biological Resources Research Article Keywords: bird assemblages, habitats, intensive practices, pesticide use, Korea Posted Date: May 27th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-551000/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Effect of Different Management Techniques on Bird Assemblages on Rice Fields in the Republic of Korea Green Choi Kyung Hee University Min Seock Do National Institute of Biological Resources Seok-Jun Son Korea Institute of Wildlife Ecology Hyung-Kyu Nam (  namhk2703@korea.kr ) National Institute of Biological Resources Introduction Rice fields are spatially monotonous [1, 2], yet they are valuable habitats for several species, including aquatic plants, benthic invertebrates, and vertebrates [1, 3]. They also provide essential alternative habitats for a range of species, including those in need of conservation, particularly due to the loss of natural habitats [4, 5]. Rice production has greatly increased in recent years due to intensive practices for maximizing crop yield, but the habitat quality and biodiversity of rice fields have been decreasing [6–10]. Intensive practices can be characterized by the excessive use of chemicals, such as herbicides and pesticides, the expansion of farming areas, and the use of advanced machinery [11]. These intensive practices have been implemented worldwide and have gravely decreased the biodiversity of various organisms, such as birds, mammals, amphibians, and invertebrates [12–16]. Rice fields can play an important role in maintaining or increasing biodiversity; however, their habitat quality is largely determined by farming methods. Rice fields can be structurally divided into paddy, levee, road, and ditch, with each component providing habitats with different ecological functions [6, 17–19]. The paddy accounts for the largest area and can support the highest richness and abundance compared with the other habitat types [20, 21]. This space can be used by various avian species depending on whether the paddy fields are flooded or dry, which varies according to the cultivation season [20, 21]. The boundary between paddies is known as the levee. It provides a stable habitat that compensates for the decreased habitat function of the paddy when the density of rice is high during the growing season, which can prevent the access of avian species [17, 22, 23]. Roads expand other habitats, such as levees and ditches, and form terrestrial ecosystems [6, 24]. Ditches are paths that allow irrigation of the paddy, form aquatic ecosystems, and shape the structures of the unique aquatic community, depending on the source of agricultural water, such as reservoirs and rivers [6, 24]. Chemical substances are intensively used in conventional rice fields, and it is well documented that they can directly kill avian species or indirectly reduce the abundance of food sources when used excessively [25–28]. Moreover, the effects of chemicals on habitat use vary among avian species [29]. In contrast, eco-friendly rice fields minimize or remove the use of pesticides. Choi et al. Research Article License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/16 Page 1/16 Abstract Many bird species rely on the ecological functions of rice field microhabitats. However, recent intensive practices are causing rice fields to provide fewer suitable habitats. This study examined bird microhabitat usage and the effects of intensive use of chemical substances on it. Eco-friendly and conventional rice fields were surveyed from January 2014 to December 2016, and the species presence, the number of birds, and the location of observed microhabitats were recorded. It was found that shorebirds and herons used more eco-friendly rice paddies with fewer or no chemical substances, while waterfowl used the paddy microhabitat more than the other microhabitats, regardless of the amount of chemical substances used. It was confirmed that land birds used ditches or roads in conventional rice fields more than those in fields that used chemical substances. Chemical substances affected bird assemblages differently. Consequently, bird assemblages used habitats differently depending on the farm crop cultivation as well as food abundance. These results provided valuable information useful for managing rice fields that serve as habitats for birds. Results We observed a total of 64,736 individuals of 113 species: 22 shorebird species (822 individuals), 11 heron species (9,538 individuals), 15 waterfowl species (45,036 individuals), 53 land bird species (9,116 individuals), and 12 other waterbird species, including cormorants, gulls, cranes and watercock (224 individuals) (see Supplementary Table S2). The average number of individuals was the highest in October (6,973 individuals) and the lowest in June (439.3 individuals). The average number of species was the highest in May (29.7 species) and the lowest in January (15 species) (Fig. 1). We observed a total of 64,736 individuals of 113 species: 22 shorebird species (822 individuals), 11 heron species (9,538 individuals), 15 waterfowl species (45,036 individuals), 53 land bird species (9,116 individuals), and 12 other waterbird species, including cormorants, gulls, cranes and watercock (224 individuals) (see Supplementary Table S2). The average number of individuals was the highest in October (6,973 individuals) and the lowest in June (439.3 individuals). The average number of species was the highest in May (29.7 species) and the lowest in January (15 species) (Fig. 1). Pattern from the self-organizing map The 36-month survey data was divided into four clusters using a self-organizing map based on the 113 bird species observed in the eco-friendly and conventional rice fields (Fig. 2). Clusters 1 and 3 were bird assemblages observed in eco- friendly rice fields (Fig. 3; the proportion of eco-friendly rice fields, cluster 1 = 100%, cluster 3 = 80%). Clusters 2 and 4 were evenly composed of birds observed in eco-friendly rice fields and those observed in conventional rice fields (Fig. 3; cluster 2, eco-friendly rice fields = 46.88%, and eco-conventional rice fields = 53.13%; cluster 4, eco-friendly rice fields = 43.75%; and conventional rice fields = 56.25%). These clusters were significantly different (Fig. 2; multi-response permutation procedure (MRPP), A=0.14, P = 0.002). Land birds accounted for the proportion of individuals (2.77–28.95%) and species (43.96–63.80%) in all clusters (Fig. 4). In the waterbird assemblages, the proportion of individuals and species for shorebirds (29.54% and 34.07%), herons (67.33% and 36.99%), and waterfowl (95.46% and 31.68%) were highest in clusters 1, 2, and 3, respectively (Fig. 4). Introduction [30] reported that common greenshanks Tringa nebularia preferred eco-friendly paddies, while the habitat use of wood sandpipers Tringa glareola was not significantly affected by the use of chemical substances. However, previous studies have only evaluated the effects of chemicals on the paddies, and no study has examined their effects on the habitat use of avian species (i.e., paddies, levees, ditches, and roads). The objective of this study was to evaluate the effects of chemical substance usage among intensive practices (i.e., eco- friendly vs. conventional) on bird assemblages using rice field microhabitats. In particular, we aimed to examine (1) the characteristics of the bird assemblages using the paddy as a habitat and (2) the differences in the microhabitat types used by the bird assemblages in eco-friendly and conventional fields, to broaden the understanding of habitat structure and usage patterns of avian species in rice fields. Page 2/16 Page 2/16 Usage characteristics of eco-friendly and conventional rice fields We found no spatial autocorrelation in waterbird guilds, whereas the land bird guild represented spatial autocorrelation. Thus, the generalized linear mixed model (GLMM) analysis showed that the relationship between habitat type and cultivation method varied according to bird guild (Table 1). Habitat type, cultivation method, and the interaction between habitat type and cultivation method were related to distribution in shorebirds and herons. However, only habitat type was related to distribution in the waterfowl. The number of shorebirds and herons observed in eco-friendly rice fields was more than twice that observed in conventional rice fields (Fig. 5). Regardless of cultivation treatment, most shorebirds, herons, and waterfowls were observed in paddy fields, while most land birds were observed in ditches (Fig. 6). Generally, herons and waterfowl were more observed in the eco-friendly rice field than in the conventional rice field, whereas more shorebirds were observed on the levee in the conventional rice field than on that of the eco-friendly fields. More land birds were observed in all the conventional rice field microhabitats, except for the levee microhabitat (Fig. 6). Page 3/16 Page 3/16 Page 3/16 Page 3/16 Table 1 Moran's I test for spatial autocorrelation and results of generalized linear mixed models used to examine the impacts of cultivation methods and habitat types on the abundance of bird assemblages in rice fields. For analyses of the bird assemblages, the response variable is the number of individuals in each field. The explanatory variables are the habitat types (paddy, levee, ditch, and road) and practice methods (eco-friendly and conventional), and the random variable is the survey period and field location. Usage characteristics of eco-friendly and conventional rice fields The bold fonts represent statistical significant y g Bird assemblages Moran's I test Generalized linear mixed models Observed Expected SD P- value Variables Estimate SE χ2 df P- value Shorebirds -0.020 -0.008 0.017 0.490 Habitat 1.126 0.134 0.692 3 < 0.001 　　　　　 Practice methods 1.113 0.302 4.484 1 0.032 　　　　　 Habitat ⅹ Practice methods 1.613 0.290 4.497 3 0.034 　　　　　　　　　　　 Herons 0.002 -0.001 0.003 0.170 Habitat 0.037 0.147 125.970 3 < 0.001 　　　　　 Practice methods 0.451 0.125 95.798 1 < 0.001 　　　　　 Habitat ⅹ Practice methods 1.136 0.128 172.630 3 < 0.001 　　　　　　　　　　　 Waterfowl 0.006 -0.003 0.007 0.190 Habitat 0.357 0.518 36.529 3 < 0.001 　　　　　 Practice methods 2.401 0.398 2.760 1 0.10 　　　　　 Habitat ⅹ Practice methods 2.511 1.648 2.202 3 0.33 　　　　　　　　　　　 Land birds 0.012 -0.003 0.006 0.010 Habitat 1.604 0.160 189.210 3 < 0.001 　　　　　 Practice methods 1.073 0.180 0.112 1 0.74 　　　　　 Habitat ⅹ Practice methods 1.554 0.175 28.333 3 < 0.001 Discussion Discussion Moreover, aquatic and terrestrial ecosystems alternate in the paddy fields [20, 21, 23]. Due to these characteristics, various food sources for waterbirds, such as benthic organisms and down grains, are available repeatedly and periodically, maintaining frequent paddy use by waterbirds that feed on those. Land birds use roads and ditches more than paddy fields. Roads and ditches have well-developed diverse herbaceous plants, such as farm crops, reeds, and silver grass, which can be used as foraging or resting spaces [17, 18], whereas the paddy is a space for growing rice, a single crop, and has a monotonous vegetation structure [17, 18]. It has been reported that a higher biodiversity of herbaceous species, including crops, attracts more diverse taxa [3], such as bird feeding spiders, butterflies, and surface roaming insects [34, 35]. Therefore, land birds use roads and ditches that offer a more diverse habitat [3]. It is necessary to identify the detailed characteristics of each bird assemblage to understand their use of rice fields and the effects of cultivation methods. Each bird assemblage showed different usage characteristics due to the complex interactions of habitat type and cultivation method. This is closely related to the characteristics of the habitat environment, formed as a result of the interactions between habitat types and cultivation methods. The eco-friendly cultivation paddies, which are used frequently by shorebirds, have a higher diversity of benthic organisms, a potential food source, than the conventional cultivation paddies [36, 37]. Even though these paddies had similar shapes to conventional fields, shorebirds used them considerably more because of the larger amount of available food. In contrast, shorebirds used levees in conventional cultivation fields more than levees in eco-friendly cultivation fields, which could be related to accessibility. Shorebirds have a habit of foraging or resting in an open environment [38, 39]. In the area studied, the levees were managed in different ways according to the cultivation method. In eco-friendly fields, weeds were either left untouched or removed using a weeder, i.e., without a complete removal; whereas in conventional fields, herbicide was used, allowing for complete removal [40]. The levees in the conventional fields presented an open shape, which shorebirds prefer, and thus used more. Herons used the paddy, levee, and road habitats in the eco-friendly fields more than the habitats in the conventional fields. These habitats provided more abundant potential food sources for the herons than the conventional habitats [36, 41]. Discussion The bird communities observed in rice fields were divided into four assemblages (clusters), according to the similarity of species and population composition, which had clearly different usage patterns. Birds were observed year-round, and the shorebirds, herons, and waterfowl (waterbirds) showed seasonal migration characteristics. Moreover, habitat type and Page 4/16 cultivation method affected different bird assemblages differently, due to differences in ecological characteristics and microhabitat use (i.e., paddy, levee, ditch, and road). These characteristics were depended on the time of arrival of the different waterbird assemblages in the rice fields [17, 18, 23]. Shorebirds use the rice field as a stopover for a short period in May; herons obtain food during or after breeding (June–September); and waterfowl stay during the winter in the rice fields, from October to April [17, 18]. Moreover, the rice fields provide different habitat environments for each season, maximizing their usage by waterbird assemblages that arrive in each season [31]. Consequently, it is believed that bird assemblages using rice fields are clearly distinguished because of differences in the season of rice-field use and microhabitats [31]. Farmers prepare to cultivate rice in early May by watering the rice field [23, 32, 33], and shorebirds use rice fields during this period [23, 32, 33]. The amount of animal prey increases rapidly during the growing season of rice (June – September) and herons are continuously observed during this period [23, 32, 33]. From October to April, rice is harvested, and waterfowl feed on the remaining down grain, increasing their use of the rice field [23, 32, 33]. Therefore, the rice field provides a suitable habitat environment for the use of each bird assemblage at different times of the year. Interestingly, land birds use the available resources in the rice field year-round without any particular seasonality. Rice field microhabitats (paddy, levee, ditch, and road) provide unique habitat qualities, and birds use them differently according to their characteristics [6, 17–19]. All bird assemblages identified in this study used all the rice field microhabitat types, and each microhabitat fulfilled different functions for water and land birds. Waterbirds, such as shorebirds, herons, and waterfowl, mostly used paddies, whereas land birds used more frequently ditches or roads. The paddy, heavily used by waterbirds, accounts for the largest area of the rice field, and showed abrupt changes during the rice cultivation process [20, 21, 23]. Discussion This increased habitat diversity was possible using pesticides, which allowed various other farm crops to be grown. However, it does not mean that the pesticide use is important. The various habitat structures created by the crops are important, and it would be better to have a variety of habitat structures without the use of pesticides. In conclusion, this study reported that bird assemblages in rice fields differ based on the agricultural techniques used, as these affect the characteristics of the habitat. As natural wetlands disappear, paddies provide alternative habitats for various birds. Rice paddies are artificial wetlands that can be changed drastically through active human management over a short period of time, compared to natural wetlands. Therefore, it is very important to understand the ecological characteristics and structure of the paddy wetlands. The results of this study will be valuable for establishing management plans for various avian species that use rice fields. Consequently, seeking and implementing clear and systematic management plans for rice field habitats can contribute to the immediate protection and conservation of birds. Future studies are needed to confirm whether the effects of cultivation methods at the community level would also be valid at the species level. In conclusion, this study reported that bird assemblages in rice fields differ based on the agricultural techniques used, as these affect the characteristics of the habitat. As natural wetlands disappear, paddies provide alternative habitats for various birds. Rice paddies are artificial wetlands that can be changed drastically through active human management over a short period of time, compared to natural wetlands. Therefore, it is very important to understand the ecological characteristics and structure of the paddy wetlands. The results of this study will be valuable for establishing management plans for various avian species that use rice fields. Consequently, seeking and implementing clear and systematic management plans for rice field habitats can contribute to the immediate protection and conservation of birds. Future studies are needed to confirm whether the effects of cultivation methods at the community level would also be valid at the species level. Discussion Moreover, herons rarely used the ditch habitat because it was narrow and low in height, making it difficult for herons to access compared to other habitats [17, 18]. Waterfowl also used habitats in eco- friendly fields more than those in conventional fields. This could be because some paddies in the eco-friendly fields were Page 5/16 watered even after harvesting. Watered paddies are an important feeding ground for various waterfowl in winter [42–44], and they were observed concentrated around the watered paddy. Although land birds used all habitat types, they used conventional cultivation areas more than eco-friendly cultivation areas, contrary to waterbirds. Certain farm crops grown in the conventionally cultivated rice fields at the study site (cultivated using pesticides) and were not grown in the eco- friendly fields. These farm crops provided land birds with more diverse habitats in the conventional fields than in the eco- friendly fields, and they allowed land birds to forage or rest more [35]. It has previously been found that rice cultivation techniques, such as chemical substance use and crop cultivation around the paddy, could affect birds using rice fields [21, 26, 38, 42, 45, 46]. The results of this study also support these findings, as it was found that waterbirds preferred eco-friendly fields due to a large amount of potential food. It was also found that land birds preferred to use fields that cultivated a higher number of plant crop species, which provided more diverse habitats, even if chemical substances were used. This increased habitat diversity was possible using pesticides, which allowed various other farm crops to be grown. However, it does not mean that the pesticide use is important. The various habitat structures created by the crops are important, and it would be better to have a variety of habitat structures without the use of pesticides. It has previously been found that rice cultivation techniques, such as chemical substance use and crop cultivation around the paddy, could affect birds using rice fields [21, 26, 38, 42, 45, 46]. The results of this study also support these findings, as it was found that waterbirds preferred eco-friendly fields due to a large amount of potential food. It was also found that land birds preferred to use fields that cultivated a higher number of plant crop species, which provided more diverse habitats, even if chemical substances were used. Study sites This study was conducted in rice fields located in Nanjido-ri, Seokmun-myeon, Dangjin-gun, Chungcheongnam-do (37° 02'N, 126° 30'E: the midwestern region of Korea). The rice fields were reclaimed in 1979, and developed by the Large- scale Comprehensive Agricultural Development Project, with a total area of 3,904 ha. Rice has been initially cultivated using conventional methods in all areas, and since 1999, it has been cultivated using eco-friendly methods in some areas (574.2 ha). The eco-friendly and the conventional rice fields were adjacent to each other, separated by roads, and were irrigated by the same ditch (KRC Report, 2008). For this study, 446 lots of eco-friendly rice fields and 442 lots of conventional rice fields were selected. The mean lot area was 0.45 ha ± 0.07. The habitat types of rice fields were divided into paddies, levees, roads, and ditches. Cultivation activities (e.g., plowing, harrowing, and rice planting) were carried out throughout the season in this area, and some eco-friendly rice paddies were watered after rice harvesting. Levees and roads were managed differently between eco-friendly rice fields and conventional rice fields. Generally, eco-friendly management included both low-pesticide and organic farming, otherwise, they were defined as conventional management. Weeds on the levee and road of the eco-friendly rice fields were either left or physically removed using a weeder, while those on the conventional rice fields were removed using a chemical herbicide. In rice fields, levees and roads have a large area to cultivate farm crops. Sixteen types of farm crops (e.g., soybeans, sesame, corn, leeks, perilla, napa cabbage, and spinach) were cultivated on conventional levees and roads. Eco-friendly levees or roads did not grow farm crops even though some rice fields used pesticides. The ditches were Page 6/16 Page 6/16 made of concrete, but some ditches, such as drainage ditches, were made from soil in both eco-friendly and conventional rice fields. Reeds (Phragmites australis) or silver grasses (Miscanthus sinensis) were grown in ditches made from soil. Although some eco-friendly rice fields used pesticides (tiadinil), most of them did not (about 20 lots). Conventional rice fields use pesticides containing tiadinil, clothianidin, pyrazosulfuron-ethyl, or fentrazmide in all areas (Supplementary Table S1). Bird and habitat survey Birds were surveyed once a month for three years, from January 2014 to December 2016. The survey was conducted between 6 a.m. and noon while visiting eco-friendly rice fields (446 lots) and conventional rice fields (442 lots); alternatively, along the road; and all birds observed on the left and right sides were recorded. To minimize the misidentification of birds, the observation area was limited to one lot (approximately 100 m in length) adjacent to the road, A total of 888 surveys were conducted per month. Birds that were flying or sitting on utility poles or wires were excluded from the record, but rapacious birds and swallows (Hirundo rustica) were recorded even if they were flying. The recorded birds were divided into waterbird guilds and land bird guilds based on field observations and habitat use obtained from previous studies [17]. Waterbirds were further subdivided into shorebirds, herons, and waterfowl. The habitat type of the flying swallows was recorded as a paddy. For rapacious birds, the habitat used for food acquisition or resting was recorded as the habitat type. To minimize the bias due to the weather, the survey was not conducted on rainy or snowy days. Acknowledgments We thank the landowners who allowed us to work in their rice fields. Data availability All collected data are provided in the supplementary information or available upon request directed to corresponding authors. Author contributions Conceived of or designed study (GC, SJS and HKN); Performed research (GC, MSD, SJS and HKN); Analyzed data (HKN, MSD and GC); Contributed new methods or models (GC and HKN); Wrote the paper (GC, MSD, SJS and HKN). Competing interests The authors declare no competing interests. Statistical analysis This study used a self-organizing map, an artificial neural network using unsupervised learning, to identify the characteristics of bird assemblages observed for three years from 2014 to 2016 [47]. The data (number of individuals: 113 species × 2 cultivation methods × 36 surveys) were assigned to the input layer of the self-organization map after log- transformation (log(1+number of individuals)) due to the differences in the number of individuals of each species. The number of output layer neurons was determined by the heuristic rule of 5√n as suggested by Vesanto et al. [48], where n is the number of data points in the input layer (72 surveys). The final model generated 42 output neurons (six horizontal and seven vertical). After self-learning of the self-organizing map, neurons were classified into groups based on the similarity of each neuron. The distance between neurons was measured by the Euclidean distance, and groups were classified using the Ward linkage method [49]. An MRPP was performed to evaluate significant differences between groups. A generalized linear mixed model (GLMM, Poisson distribution, and log link) was used to evaluate the effects of habitat type (i.e., paddy, levee, ditch, and road) and cultivation methods (eco-friendly and conventional) on the number of birds per bird guild. Spatial autocorrelation analysis was conducted to examine the spatial characteristics of the data before applying the GLMM. When the result of the spatial autocorrelation analysis was significant, a GLMM analysis was conducted by applying spatial autocorrelation. In the GLMM, the number of birds per bird guild was designated as a response variable, and habitat type and cultivation method were considered as fixed effects. The timing of the survey and the location of each lot were treated as random effects. All analyses were performed using R statistical software V 3.6.1 [50]. The “kohonen” [51], the “vegan” [52], the “lme4” [53], and the “spam” packages [54] were used for the self-organizing map analysis, MRPP, GLMM without autocorrelation, and GLMM with autocorrelation, respectively. Page 7/16 References 10.1016/j.agee.2010.01.015 12. Donal, P. F., Gree, R. E. & Heath, M. F. Agricultural intensification and the collapse of Europe's farmland bird populations. Proc. Biol. Sci. 268(1462), 25-29 (2001). 10.1098/rspb.2000.1325, Pubmed:12123294 12. Donal, P. F., Gree, R. E. & Heath, M. F. Agricultural intensification and the collapse of Europe's farmland bird populations. Proc. Biol. 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The impact of agricultural activities on habitat use by the Wood sandpiper and Common greenshank in rice fields. Ornithol. Sci. 20(1), 27-37 (2021) Page 9/16 Page 9/16 31. Lourenço, P. M. & Piersma, T. Stopover ecology of Black-tailed Godwits Limosa limosa in Portuguese rice fields: a guide on where to feed in winter. Bird Study 55(2), 194-202 (2008). 10.1080/00063650809461522 32. References Fujioka, M., Lee, S. D., Kurechi, M. & Yoshida, H. Bird use of rice fields in Korea and Japan. Waterbirds 33(sp1)(sp1), 8-29 (2010). 10.1675/063.033.s102 33. Stafford, J. D., Kaminski, R. M. & Reinecke, K. J. Avian foods, foraging and habitat conservation in world rice fields. Waterbirds 33(sp1), 133-150 (2010). 10.1675/063.033.s110 34. Harwood, J. D., Sunderland, K. D. & Symondson, W. O. C. Living where the food is: web location by linyphiid spiders in relation to prey availability in winter wheat. J. Appl. Ecol. 38(1), 88-99 (2001). 10.1046/j.1365-2664.2001.00572.x 35. Morris, A. 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Testing Environmental and Genetic Effects in the Presence of Spatial Autocorrelation. Ecography. 37, 781–790 (2014). 10.1111/ecog.00566 Figures Page 11/16 Page 11/16 Figure 1 Seasonal change in the average number of (a) individuals and (b) species of birds in the rice fields of the midwestern part of South Korea. The error bars represent the standard deviation. p Figure 2 Classification of the input data (number of individuals organizing map. (a) The 42 output neurons are arrang grouped into four clusters, which are shown in differe method. The circles in the neurons indicate the input d Figure 2 Classification of the input data (number of individuals: 113 species × 2 cultivation methods × 36 surveys) using the self organizing map. (a) The 42 output neurons are arranged in a two-dimensional grid (7 × 6; V1~V42). (b) Neurons are grouped into four clusters, which are shown in different colors, using dendrograms produced using Ward’s linkage method. The circles in the neurons indicate the input data. Figure 2 Classification of the input data (number of individuals: 113 species × 2 cultivation methods × 36 surveys) using the self- organizing map. (a) The 42 output neurons are arranged in a two-dimensional grid (7 × 6; V1~V42). (b) Neurons are grouped into four clusters, which are shown in different colors, using dendrograms produced using Ward’s linkage method. The circles in the neurons indicate the input data. Page 12/16 Page 12/16 Page 12/16 Page 12/16 Figure 3 The difference in practiced method among four clusters defined by the self-organizing map. Figure 3 Figure 3 Figure 3 The difference in practiced method among four clusters defined by the self-organizing map. The difference in practiced method among four clusters defined by the self-organizing map. Page 13/16 Figure 4 The difference in the relative number of (a) individuals and (b) species of bird assemblages among the four clusters defined by the self-organizing map (SOM). Figure 6 Average values of bird group abundance ((a) shorebirds, (b) herons, (c) waterfowl, and (d) land birds) in eco-friendly and conventional fields among habitat types (paddy, levee, ditch, and road). N=36 for each farming method. Figure 4 Figure 4 The difference in the relative number of (a) individuals and (b) species of bird assemblages among the four clusters defined by the self-organizing map (SOM). The difference in the relative number of (a) individuals and (b) species of bird assemblages among the four clusters defined by the self-organizing map (SOM). The difference in the relative number of (a) individuals and (b) species of bird assemblages among the four clusters defined by the self-organizing map (SOM). Page 14/16 Figure 5 Average values of bird group abundance ((a) shorebirds, (b) herons, (c) waterfowl, and (d) land birds) between eco- friendly fields (EFF) and conventional fields (CF) according to main occurrence period (shorebirds: April-May, and August- September; herons: May-October; waterfowl: October-April; land birds: all seasons). The number above the bar graph indicates the sample size of the bird group. N=36 for each farming method. Figure 5 Average values of bird group abundance ((a) shorebirds, (b) herons, (c) waterfowl, and (d) land birds) between eco- friendly fields (EFF) and conventional fields (CF) according to main occurrence period (shorebirds: April-May, and August- September; herons: May-October; waterfowl: October-April; land birds: all seasons). The number above the bar graph indicates the sample size of the bird group. N=36 for each farming method. Page 15/16 Figure 6 Average values of bird group abundance ((a) shorebirds, (b) herons, (c) waterfowl, and (d) land birds) in eco-friendly and conventional fields among habitat types (paddy, levee, ditch, and road). N=36 for each farming method. Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. This is a list of supplementary files associated with this preprint. Click to download. Supplementaryinformation.docx Supplementaryinformation.docx Page 16/16	32,885
https://openalex.org/W1485768591	OpenAlex	Open Science	CC-By	2,013	First records of Chthonerpeton arii Cascon and Lima-Verde, 1994 (Amphibia: Gymnophiona: Typhlonectidae) out of the type locality	Adriano O. Maciel	English	Spoken	1,186	2,351	First records of Chthonerpeton arii Cascon and Lima-Verde, 1994 (Amphibia: Gymnophiona: Typhlonectidae) out of the type locality Notes on Geographic Distribution Adriano Oliveira Maciel 1, Bruno Vilela de Moraes e Silva 2,3, Filipe Augusto Cavalcanti do Nascimento Diva Maria Borges-Nojosa 4 and Daniel Cassiano Lima 4 1 Museu Paraense Emílio Goeldi, Departamento de Zoologia. Avenida Perimetral, 1901, Terra Firme. CEP: 66077-530. Belém, Pará, Brazil. 2 Universidade Federal de Alagoas, Museu de História Natural, Setor de Zoologia. Av. Aristeu de Andrade, 452, Farol. CEP: 57051-090. Maceió, 1 Museu Paraense Emílio Goeldi, Departamento de Zoologia. Avenida Perimetral, 1901, Terra Firme. CEP: 66077-530. Belém, Pará, Brazil. 2 Universidade Federal de Alagoas Museu de História Natural Setor de Zoologia Av Aristeu de Andrade 452 Farol CEP: 57051 090 Macei 1 Museu Paraense Emílio Goeldi, Departamento de Zoologia. Avenida Perimetral, 1901, Terra Firme. CEP: 66077-530. Belém, Pará, Brazil. 2 Universidade Federal de Alagoas, Museu de História Natural, Setor de Zoologia. Av. Aristeu de Andrade, 452, Farol. CEP: 57051-090. Macei Alagoas Brazil Alagoas, Brazil. 3 Universidade Federal de Goiás, Instituto de Ciências Biológicas (Bloco ICB IV), Programa de Pós-Graduação em Ecologia e Evolução, Campus II/ UFG. CEP: 74001-970. Goiânia, Goiás, Brazil. 4 Programa de Pós-Graduação em Ecologia e Recursos Naturais, Depa Graduação em Ecologia e Recursos Naturais, Departamento de Biologia, 4 Programa de Pós-Graduação em Ecologia e Recursos Natu Universidade Federal do Ceará, Fortaleza, Ceará, Brazil. Universidade Federal do Ceará, Fortaleza, Ceará, Brazil. Corresponding author. E-mail: aombiologo@yahoo.com.br Abstract: Chthonerpeton arii was described from a large series of specimens from Limoeiro do Norte municipality, state of Ceará, Brazil. Here we provide the first records of the species out of the type locality, in the state of Bahia, and in its border with the state of Pernambuco, Brazil. We also provide color photographs of a preserved specimen. distribution in Brazil, occurring in water bodies in the Caatinga biome: temporary ponds at the type locality; in a river and in a fish breeding station in our new locality records. Collection of more specimens will be needed to improve the knowledge of it species. Typhlonectidae Taylor, 1968 is a family of Gymnophiona, which includes the aquatic and semiaquatic caecilians of South America (Taylor 1968). There are five recognized genera up till now, for which Chthonerpeton Peters, 1880 is the most speciose, with eight known species (Wilkinson and Nussbaum 2006). Chthonerpeton arii Cascon and Lima-Verde, 1994 was described based on 60 specimens collected in the municipality of Limoeiro do Norte, state of Ceará, northeastern Brazil. Journal of species lists and distribution Chec List Journal of species lists and distribution Chec List Check List 9(4): 818–819, 2013 © 2013 Check List and Authors ISSN 1809-127X (available at www.checklist.org.br) Notes on Geographic Distribution Wilkinson, M. and R.A. Nussbaum. 2006. Caecilian phylogeny and classification; pp. 39-78 In J. M. Exbrayat (Ed.). Reproductive Biology and phylogeny of Gymnophiona (caecilians). Enfield: Science Publisher Inc. First records of Chthonerpeton arii Cascon and Lima-Verde, 1994 (Amphibia: Gymnophiona: Typhlonectidae) out of the type locality Here, we report the first records of this species out of the type locality, in the state of Bahia and in its border with the state of Pernambuco, in the São Francisco river (Figure 1). Figure 1. Distribution of Chthonerpeton arii. Star = type locality, municipality of Limoeiro do Norte, state of Ceará, Brazil; upper circle = UHE Itaparica, border between states of Bahia and Pernambuco, Brazil (approx. 09°08’ S; 38°18’ W; 270 m elev.); lower circle = municipality of Paulo Afonso, state of Bahia, Brazil (approx. 09° 22’ S; 38° 13’W; 235 m elev.). g We found three specimens of C. arii in two Brazilian herpetological collections. One male specimen in the Universidade Federal do Ceará (UFC), Fortaleza, state of Ceará (UFC A2465; 282 mm total length; collected by J. S. Lima-Verde; September 1988) from the hydroelectric of Itaparica (current hydroelectric of Luiz Gonzaga), on the border between the states of Pernambuco and Bahia; and two female specimens in the Herpetological collection of the Museu de História Natural of Universidade Federal de Alagoas (MUFAL), Maceió, state of Alagoas (MUFAL 8363-64; respectively, 385 mm and 457 mm total length, Figure 2; collected by CHESF officials; July 1982) from a fish breeding station in the municipality of Paulo Afonso, state of Bahia. These two new localities are approximately 30 km apart from each other, and 440 km and 470 km, respectively, from the type locality. The species identification was made by comparison to the type series. Specimens from Paulo Afonso are more robust than the other known specimens of C. arii but we do not have an explanation about, since we have few specimens from it locality to test geographic variation. Chthonerpeton arii is the species in the genus with the most septentrional Specimens from Paulo Afonso are more robust than the other known specimens of C. arii but we do not have an explanation about, since we have few specimens from it locality to test geographic variation. Chthonerpeton arii is the species in the genus with the most septentrional Figure 1. Distribution of Chthonerpeton arii. Star = type locality, municipality of Limoeiro do Norte, state of Ceará, Brazil; upper circle = UHE Itaparica, border between states of Bahia and Pernambuco, Brazil (approx. 09°08’ S; 38°18’ W; 270 m elev.); lower circle = municipality of Paulo Afonso, state of Bahia, Brazil (approx. 09° 22’ S; 38° 13’W; 235 m elev.). First records of Chthonerpeton arii Cascon and Lima-Verde, 1994 (Amphibia: Gymnophiona: Typhlonectidae) out of the type locality 818 Maciel et al. \| First records of Chthonerpeton arii out of the type locality Maciel et al. \| First records of Chthonerpeton arii out of the type locality Figure 2. Specimen of Chthonerpeton arii (MUFAL 8364). Lateral (A), dorsal (B), and ventral (C) view of the head; cloacal opening, and (D) entire specimen (scale = 3 cm). (Photos by Bruno Vilela). Figure 2. Specimen of Chthonerpeton arii (MUFAL 8364). Lateral (A), dorsal (B), and ventral (C) view of the head; cloacal opening, and (D) entire specimen (scale = 3 cm). (Photos by Bruno Vilela). Figure 2. Specimen of Chthonerpeton arii (MUFAL 8364). Lateral (A), dorsal (B), and ventral (C) view of the head; cloacal opening, and (D) entire specimen (scale = 3 cm). (Photos by Bruno Vilela). Acknowledgments: We are grateful to Gabriel Skuk (in memorian) who called us attention about these specimens. Acknowledgments: We are grateful to Gabriel Skuk (in memorian) who called us attention about these specimens. Acknowledgments: We are grateful to Gabriel Skuk (in memorian) who called us attention about these specimens. Received: August 2012 Accepted: May 2013 Published online: August 2013 Editorial responsibility: Raúl Maneyro Literature Cited Literature Cited Cascon, P. and J.S. Lima-Verde. 1994. Uma nova espécie de Chthonerpeton do nordeste brasileiro (Amphibia, Gymnophiona, Typhlonectidae). Revista Brasileira de Biologia 54(4): 549-553. Silva, H.R., M.C. de Britto-Pereira and U. Caramaschi. 2003. A new species of Chthonerpeton (Amphibia: Gymnophiona: Typhlonectidae) from Bahia, Brazil. Zootaxa 381: 1-11. Received: August 2012 Accepted: May 2013 Published online: August 2013 Editorial responsibility: Raúl Maneyro Taylor, E.H. 1968. The Caecilians of the World. A taxonomic review. University of Kansas Press. Lawrence, Kansas. 848 pp. 819	9,207
https://openalex.org/W4281640800	OpenAlex	Open Science	CC-By	2,022	Early neurological deterioration in acute ischemic stroke patients after intravenous thrombolysis with alteplase predicts poor 3-month functional prognosis - data from the Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China (TIMS-China)	Fengli Che	English	Spoken	4,097	8,972	Early Neurological Deterioration in Acute Ischemic Stroke Patients after Intravenous Thrombolysis with Alteplase Predicts Poor 3-Month Functional Prognosis - Data from the Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China (TIMS-China) Fengli Che Beijing Tiantan Hospital, Capital Medical University Anxin Wang Beijing Tiantan Hospital, Capital Medical University Yi Ju Beijing Tiantan Hospital, Capital Medical University Yarong Ding Beijing Luhe Hospital, Capital Medical University Honglian Duan Beijing Luhe Hospital, Capital Medical University Xiaokun Geng Beijing Luhe Hospital, Capital Medical University Xingquan Zhao (  zxq@vip.163.com ) Beijing Tiantan Hospital, Capital Medical University Yongjun Wang Beijing Tiantan Hospital, Capital Medical University Research Article Page 1/12 Keywords: acute ischemic stroke, intravenous thrombolysis, early neurological deterioration, prognosis Posted Date: February 11th, 2022 DOI: https://doi.org/10.21203/rs.3.rs-1337645/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Posted Date: February 11th, 2022 Page 1/12 Page 1/12 Page 1/12 Introduction Stroke has become one of the leading causes of death and disability of humans(1), and there has been a high incidence of stroke in China(2). It has been confirmed that target-vessel revascularization is the most effective method to reduce the disability and mortality of patients. Meanwhile, intravenous thrombolysis with recombinant tissue-type plasminogen activator (rt-PA) has been the most economical and convenient treatment(3). However, some studies have found that some patients still suffered severe neurological deterioration after receiving intravenous thrombolysis, which resulted in prolonged hospitalization and severe adverse prognosis(4). This study explored the risk factors of early neurological deterioration (END) and the correlation between END and 3-month functional prognosis. Abstract Background： We aimed to investigate the risk factors of early neurological deterioration (END) after intravenous thrombolysis with recombinant tissue-type plasminogen activator (rt-PA) and the relationship between END and poor 3-month functional outcomes. Methods: Patients who accepted intravenous recombinant rt-PA were enrolled continuously. END was defined as an increase of National Institute of Health Stroke (NIHSS) score ≥ 4 points or death within 24 hours after intravenous thrombolysis. The modified Rankin Scale (mRS) score was recorded to evaluate the functional prognosis of stroke, and the poor 3-month prognosis was defined as an mRS score ≥ of 3. Univariate and multivariate analyses were used to analyze the risk factors of END. The relation between END and 3-month functional outcome was analyzed by multivariate logistic regression analysis. Results: A total of 1107 patients (mean age, 63.42±11.33 years; 673 males) were included in the final analysis, and 81(7.32%) patients had END. In multivariate analysis, the serum glucose level was significantly associated with END; the odds ratio was 1.10 (95% CI 1.03 to 1.18, p =0.004). The multivariate logistic analysis showed END has a notable association with the poor 3-month functional recovery even after adjusting for confounding factors; the adjusted OR was 8.25 (95% CI 3.77 to 18.03, p<0.0001). Conclusions: The initial serum glucose level might be an independent risk factor of END, and END might predict a poor 3-month prognosis. Definition of END and Clinical outcome measurement END was defined as an increase of NIHSS (National Institute of Health Stroke) score ≥ 4 points or death within 24 hours after intravenous thrombolysis(8). The primary outcome was poor 3-month functional recovery, expressed as a modified Rankin Scale (mRS) score ≥3. The secondary outcomes were sICH (symptomatic intracranial hemorrhage) and mortality at seven days and 90 days. We used the definitions of sICH in the following three studies: Safe Implementation of Thrombolysis in Stroke-Monitoring Study (SITS-MOST)(8), National Institute of Neurological Disorders and Stroke (NINDS)(9), and European Cooperative Acute Stroke Study Ⅱ (ECASS Ⅱ)(10). Statistical analysis Continuous variables were described by means (standard deviations [SDs]) or medians (interquartile ranges [IQRs]). Categorical variables were presented as frequencies and percentages. The baseline characteristics of patients between the END group and the non-END group were compared by Wilcoxon rank-sum tests for continuous variables and X2 test for categorical variables. Univariate and multivariate logistic regression was used to estimate the odds ratios (ORs), the corresponding 95% confidence intervals (CIs), and the adjusted ORs with their 95% CI. The multiple ordinal regression was used to test the distribution of mRS at 3-month of patients. SAS software performed all statistical analyses, version 9.4 (SAS Institute Inc., Cary, NC, USA). All P values were two-sided, with P < 0.05 considered statistically significant. Study Population The data were obtained from the Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China (TIMS-CHINA) database - a multicenter prospective stroke registry program that enrolled patients who received intravenous tPA within 4.5hours after symptom onset from May 2007 to July 2012 in China(5). Some previous pieces of literature have reported the trial design and some results of the Page 2/12 study(6, 7). The ethics committee approved the study protocol of Beijing Tiantan Hospital with the Helsinki Declaration. The quality monitoring committee of TIMS-China and the Contract Research Organization independently have been regularly monitoring the registry. All participants had signed written consent. Baseline Characteristics A total of 1107 consecutive patients (mean age, 63.42±11.33 years; 673 males) were included in the final analysis, among which 81(7.32%) patients occurred END (Figure 1). Between the END group and the non- END group, there were statistical differences in the history of prior stroke/TIA (2.47% vs.9.75%, p=0.029), initial serum glucose level (9.00±4.35 mmol/L vs.7.58±2.87 mmol/L, p=0.001), fibrinogen (3.47±1.24 g/L vs. 3.23±1.23 g/L, p=0.040), low-density lipoprotein (3.20±0.90 mmol/L vs. 2.92±0.96 mmol/L, p=0.003), cholesterol (5.08±1.16 mmol/L vs. 4.85±1.20 mmol/L, p=0.021), SBP (systolic blood pressure) on admission (152.30 ±18.56 mmHg vs. 147.57±21.09 mmHg, p=0.039), DBP (diastolic blood pressure) on admission (88.35±11.86 mmHg vs. 85.70±12.68 mmHg, p=0.038), taking aspirin within seven days before thrombolysis (38.27% vs. 65.20, p<0.0001), and taking clopidogrel within seven days before thrombolysis(13.58% vs. 23.59%, p=0.039). There was no significant statistical difference in the neurological deficit on admission between the two groups. Concerning TOAST types, although the Page 3/12 Page 3/12 proportion of CE in the END group was higher than in the non-END group (28.21% vs. 18.69%), there was no difference in the etiology distribution between the two groups. In multivariate analysis, END has a significant correlation with the initial serum glucose level (OR,1.10, 95%CI 1.03-1.18; p=0.004), taking aspirin within seven days before thrombolysis (OR, 0.25, 95%CI 0.14-0.44; p<0.0001), and taking clopidogrel within seven days before thrombolysis (OR,0.39, 95%CI 0.19-0.82; p=0.013). The demographics and clinical characteristics at the baseline of subjects in this study were demonstrated in Table 1, and multivariate logistic regression analysis for risk factors of END was shown in Table 2. Clinical outcomes During the follow-up, 23 patients were excluded because of missing, and there were 1084 patients (97.92%) who had a 3-month mRS score. The proportion of poor function outcomes is 83.54% in the END group and 37.41% in the non-END group (crude OR 8.49; 95%CI 4.62 to 15.60; P<0.0001). After adjusting the baseline variables as the history of prior stroke/TIA, initial serum glucose level, fibrinogen, low-density lipoprotein, cholesterol, SBP on admission, DBP on admission, taking aspirin within seven days before thrombolysis, taking clopidogrel within seven days before thrombolysis, and TOAST types, END has a statistical correlation with poor 3-month functional outcomes, the adjusted OR was 8.25(95%CI 3.77 -18.03; P<0.0001; Table 3). There was a numerical difference in the distribution of 3-month mRS among patients in the two groups (crude p <0.0001), and this difference was still significant after adjusting compound factors (adjusted OR 11.74, 95%CI 7.58 to 18.18; P<0.0001; Figure 2). Regarding the secondary outcomes, END has a prominent correlation with SICH (NINDS), the adjusted OR was 12.53 (95%CI 5.15 -30.49; P<0.0001; Table 3). Meanwhile, END has a significant correlation with mortality at seven days and mortality at 90 days, the adjusted OR was 20.92(95%CI 7.45 -58.72; P<0.0001; Table 3) and 8.06(95%CI 3.91 -16.62; P<0.0001; Table 3). Discussion Our study aimed to explore the risk factors of END and the relationship between END and poor 3-month functional outcomes. In some studies, the incidence of END was significantly different due to the lack of a unified definition of END, which was from 5.8% to 34.9%(11-14). At present, most studies have defined END as an increasing NIHSS score ≥ 4 points or death within 24 hours after intravenous thrombolysis(8, 15), which was the exact definition of END in our study. Our study enrolled a total of 1107 patients accepting intravenous thrombolysis, and 81 (7.32%) patients occurred END. Simonsen et al. studied a total of 569 patients who received reperfusion therapy and found the incidence of END was 5.8%(13). Concerning the risk factors of END, although some experts generally believe that elderly patients are more prone to END, it has not been confirmed in some studies(9, 14). Some studies also have demonstrated the predictors of END as follows: diabetes(16), neurological functional deficits on admission(17), and systolic BP(18-20). Page 4/12 There was no statistical difference in age between the END group and the non- END group in our study. Still, there were statistical differences in the history of prior stroke/TIA, initial serum glucose level, There was no statistical difference in age between the END group and the non- END group in our study. Still, there were statistical differences in the history of prior stroke/TIA, initial serum glucose level, Page 4/12 fibrinogen, low-density lipoprotein, cholesterol, SBP on admission, and DBP on admission between the two groups. However, in the multivariate analysis, our study found the initial serum glucose level was an independent risk factor of END; the odds ratio was 1.10 (95%CI 1.03 to 1.18, p =0.004). We have an exciting discovery that taking antiplatelets (aspirin or clopidogrel) within seven days might be the protective factor of END. The incidence of END in patients taking aspirin could be 0.25 times lower than patients without aspirin, similar to patients taking clopidogrel within seven days. Due to the limitation of our research, we had no further study on this result, and it would be a meaningful research focus in the future. The series of pathophysiological reactions of the brain after intravenous thrombolysis such as intracranial hemorrhage (ICH)(21, 22), malignant edema(23), early recurrent ischemic stroke(11), and early seizures(24) resulted in the aggravation of neurological deficit(18, 19). Meanwhile, SICH had been the leading cause of END. Abbreviations TIA, transient ischemic attack; SBP, systolic blood pressure; DBP, diastolic blood pressure; WBC, white blood cell; PLT, platelet; INR, international normalized ratio; PT, prothrombin time; APTT, activated partial thromboplastin time; FBG, fibrinogen; NIHSS, National Institute of Health Stroke; LDL, low-density lipoprotein; TC, cholesterol; IQR, interquartile ranges; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large-artery atherosclerosis; Scale; SAO, small-artery occlusion; CE, cardioembolism; OR, odds ratio; CI, confidence interval; TIA, transient ischemic attack; SBP, systolic blood pressure; DBP, diastolic blood pressure; FBG, fibrinogen; LDL, low-density lipoprotein; TC, cholesterol; mRS, modified Rankin Scale; SICH, symptomatic intracranial hemorrhage; SITS-MOST, safe implementation of treatments in stroke- monitoring; ECASS II, second European–Australasian acute stroke study; NINDS, National Institute of Neurological Disorders and Stroke. Conclusion Early neurological deterioration has a high incidence after intravenous thrombolysis and the initial serum glucose level might be an independent risk factor of END. END might predict a poor 3-month prognosis. It might be essential to understand the underlying mechanism of END. Discussion Our study confirmed that the patients in the END group had a higher incidence of ICH than in the non-END group. The incidence of SICH(NINDS) in the END group patients could be 12.53 times higher than patients in the non-END group. Furthermore, END was significantly correlated with mortality at seven days and mortality at three months. Compared the patients with non- END, the mortality at seven days of patients with END could be 20.92 times higher, and the mortality at three months could be 8.06 times higher. We gratefully appreciate all the participants and staff for their contributions. We gratefully appreciate all the participants and staff for their contribution Consent for publication Not applicable. Funding The study was supported by grants from the Beijing Natural Science Foundation (Z200016). Declarations Ethics approval and consent to participate The ethics committee of Beijing Tiantan Hospital approved this study. All study subjects had signed informed consent. We confirm that all methods in our study were performed in accordance with the relevant guidelines and regulations. Competing interests The authors declare that they have no competing interests. Declarations Page 5/12 Authors’ contributions Concept and design: FC, XZ, XG, and YJ. Drafting of the manuscript: FC, YL, and XG. Critical revision of the manuscript for important intellectual content: HD, YD, and JJ. Provision of study material or patients: YL and YJ. Collection and assembly of data: FC, YL, JJ, AW, HD, YJ, YD, and XG. Check and approve of clinical definition: XG and XZ. Data analysis: FC and AW. Data interpretation: FC, AW, HD, JJ, YD, and XG. Administrative, technical or material support: XZ and YJ. Supervision: XG and XZ. Final approval of manuscript: all authors. Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. References 1. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016: a systematic analysis for the Global Burden of Disease Study 2016. LANCET. 2017;390(10100):1151- 210. doi:10.1016/s0140-6736(17)32152-9 1. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016: a systematic analysis for the Global Burden of Disease Study 2016. LANCET. 2017;390(10100):1151- 210. doi:10.1016/s0140-6736(17)32152-9 2. Wu S, Wu B, Liu M, et al. Stroke in China: advances and challenges in epidemiology, prevention, and management. Lancet Neurol. 2019;18(4):394-405. doi:10.1016/s1474-4422(18)30500-3 2. Wu S, Wu B, Liu M, et al. Stroke in China: advances and challenges in epidemiology, prevention, and management. Lancet Neurol. 2019;18(4):394-405. doi:10.1016/s1474-4422(18)30500-3 2. Wu S, Wu B, Liu M, et al. Stroke in China: advances and challenges in epidemiology, prevention, and management. Lancet Neurol. 2019;18(4):394-405. doi:10.1016/s1474-4422(18)30500-3 Page 6/12 3. Marko M, Posekany A, Szabo S, et al. Trends of r-tPA (Recombinant Tissue-Type Plasminogen Activator) Treatment and Treatment-Influencing Factors in Acute Ischemic Stroke. STROKE. 2020;51(4):1240-7. doi:10.1161/strokeaha.119.027921 4. Thiebaut AM, Gauberti M, Ali C, et al. The role of plasminogen activators in stroke treatment: fibrinolysis and beyond. Lancet Neurol. 2018;17(12):1121-32. doi:10.1016/s1474-4422(18)30323-5 5. Liao XL, Wang CX, Wang YL, et al. Implementation and outcome of thrombolysis with alteplase 3 to 4. 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Blood pressure variability and the development of early neurological deterioration following acute ischemic stroke. J Hypertens. 2015;33(10):2099-106. doi:10.1097/hjh.0000000000000675 19. Park TH, Lee JK, Park MS, et al. Neurologic deterioration in patients with acute ischemic stroke or transient ischemic attack. NEUROLOGY. 2020;95(16):e2178-e91. doi:10.1212/wnl.0000000000010603 20. Gong P, Zhou J, Zhang Y. Letter by Gong et al Regarding Article, "Predictors of Unexplained Early Neurological Deterioration After Endovascular Treatment for Acute Ischemic Stroke". STROKE. 2021;52(1):e44-e5. doi:10.1161/strokeaha.120.032713 21. Mori M, Naganuma M, Okada Y, et al. Early neurological deterioration within 24 hours after intravenous rt-PA therapy for stroke patients: the Stroke Acute Management with Urgent Risk Factor Assessment and Improvement rt-PA Registry. Cerebrovasc Dis. 2012;34(2):140-6. doi:10.1159/000339759 22. Wei XE, Zhao YW, Lu J, et al. Timing of recanalization and outcome in ischemic-stroke patients treated with recombinant tissue plasminogen activator. Acta Radiol. 2015;56(9):1119-26. doi:10.1177/0284185114545151 23. Battey TW, Karki M, Singhal AB, et al. Brain edema predicts outcome after nonlacunar ischemic stroke. STROKE. 2014;45(12):3643-8. doi:10.1161/strokeaha.114.006884 23. Battey TW, Karki M, Singhal AB, et al. Brain edema predicts outcome after nonlacunar ischemic stroke. STROKE. 2014;45(12):3643-8. doi:10.1161/strokeaha.114.006884 24. Jung S, Schindler K, Findling O, et al. Adverse effect of early epileptic seizures in patients receiving endovascular therapy for acute stroke. STROKE. 2012;43(6):1584-90. doi:10.1161/strokeaha.111.645358 24. Jung S, Schindler K, Findling O, et al. Adverse effect of early epileptic seizures in patients receiving endovascular therapy for acute stroke. STROKE. 2012;43(6):1584-90. doi:10.1161/strokeaha.111.645358 Tables Table 1 Demographic and clinical characteristics in patients Page 8/12 Page 8/12 Variables END (n =81) non-END (n = 1026) P-Value Gender, male, n (%) 49(60.49) 624(60.82) 0.954 Age, mean ±SD (years) 64.47±9.34 63.34±11.48 0.636 Hypertension, n (%) 54(66.67) 598(58.34) 0.143 Diabetes mellitus, n (%) 17(20.99) 174(16.96) 0.356 Atrial fibrillation, n (%) 15(18.52) 182(17.74) 0.860 Hyperlipidemia, n (%) 3(3.70) 69(6.73) 0.288 prior stroke/TIA, n (%) 2(2.47) 100(9.75) 0.029 Current smoking, n (%) 20(24.69) 361(35.19) 0.056 Glucose, mean (SD), mmol/L 9.00±4.35 7.58±2.87 0.001 WBC, mean (SD), ×109/L 8.32±2.85 7.87±2.70 0.216 FBG, mean (SD), g/L 3.47±1.24 3.23±1.23 0.040 LDL, mean (SD), mmol/L 3.20±0.90 2.92±0.96 0.003 TC, mean (SD), mmol/L 5.08±1.16 4.85±1.20 0.021 SBP on admission, mean (SD), mmHg 152.30±18.56 147.57±21.09 0.039 DBP on admission, mean (SD), mmHg 88.35±11.86 85.70±12.68 0.038 SBP at 2 hours after thrombolysis, mean (SD), mmHg 150.05 ±22.31 143.03±19.80 0.002 DBP at 2 hours after thrombolysis, mean (SD), mmHg 87.19±14.28 83.42±12.38 0.015 NIHSS on admission, IQR 12(3-28) 11(0-40) 0.559 Pre-admission mRS 0–2, n (%) 80(100) 1002(92.75) 0.398 Baseline medication within seven days before thrombolysis Taking aspirin, n (%) 31(38.27) 669(65.20) <0.0001 Taking clopidogrel, n (%) 11(13.58) 242(23.59) 0.039 Taking other antiplatelets, n (%) 11(13.58) 156(15.20) 0.694 Taking hypoglycemic drugs , n (%) 0(0.00） 9(0.88) 1.000 Median door-to-needle time, IQR, h 1.82±0.95 2.60±9.90 0.971 TOAST bt 0 080 Page 9/12 Page 9/12 LAA, n (%) 43(55.13) 553(54.11) SAO, n (%) 4(5.13) 113(11.06) CE, n (%) 22(28.21) 191(18.69) Other, n (%) 9(11.54) 165(16.14) Abbreviation: TIA, transient ischemic attack; SBP, systolic blood pressure; DBP, diastolic blood pressure; WBC, white blood cell; PLT, platelet; INR, international normalized ratio; PT, prothrombin time; APTT, activated partial thromboplastin time; FBG, fibrinogen; NIHSS, National Institute of Health Stroke; LDL, low-density lipoprotein; TC, cholesterol; IQR, interquartile ranges; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large-artery atherosclerosis; Scale; SAO, small-artery occlusion; CE, cardioembolism. LAA, n (%) 43(55.13) 553(54.11) SAO, n (%) 4(5.13) 113(11.06) CE, n (%) 22(28.21) 191(18.69) Other, n (%) 9(11.54) 165(16.14) Abbreviation: TIA, transient ischemic attack; SBP, systolic blood pressure; DBP, diastolic blood pressure; WBC, white blood cell; PLT, platelet; INR, international normalized ratio; PT, prothrombin time; APTT, activated partial thromboplastin time; FBG, fibrinogen; NIHSS, National Institute of Health Stroke; LDL, low-density lipoprotein; TC, cholesterol; IQR, interquartile ranges; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large-artery atherosclerosis; Scale; SAO, small-artery occlusion; CE, cardioembolism. Tables (%) of patients Unadjusted OR (95% CI) P value Adjusted OR (95% CI) * P value END group (n=81) non-END group (n=1026) Primary outcome mRS 3–6 at three months 66(83.54) 376(37.41) 8.49 (4.62-15.60) <0.0001 8.25 (3.77- 18.03) <0.0001 Safety outcomes SICH(SITS-MOST) 15(18.52) 1(0.10) 232.83(-) <0.0001 109.77(-） 0.001 SICH (ECASS II) 30(37.04) 5(0.49) 120.12 (44.74- 322.50) <0.0001 90.46 (19.84- 412.44) <0.0001 SICH (NINDS) 30(37.04) 22(2.14) 26.84 (14.47- 49.79) <0.0001 12.53 (5.15- 30.49) <0.0001 Mortality at seven days 26(32.50) 16(1.56) 30.39 (15.39- 60.01) <0.0001 20.92 (7.45- 58.72) <0.0001 Mortality at three months 36(45.00) 59(5.86) 13.13 (7.86-21.94) <0.0001 8.06 (3.91- 16.62) <0.0001 Adjusted baseline variables: prior stroke/TIA, Systolic BP before thrombolysis, Diastolic BP before thrombolysis, glucose before thrombolysis, FBG before thrombolysis, Systolic BP at 2 hours after thrombolysis, Diastolic BP at 2 hours after thrombolysis, LDL after thrombolysis, TC after thrombolysis, Aspirin use within seven days after thrombolysis, clopidogrel use within seven days after thrombolysis, b Adjusted baseline variables: prior stroke/TIA, Systolic BP before thrombolysis, Diastolic BP before thrombolysis, glucose before thrombolysis, FBG before thrombolysis, Systolic BP at 2 hours after thrombolysis, Diastolic BP at 2 hours after thrombolysis, LDL after thrombolysis, TC after thrombolysis, Aspirin use within seven days after thrombolysis, clopidogrel use within seven days after thrombolysis, OCSP subtypes. *Adjusted baseline variables: prior stroke/TIA, Systolic BP before thrombolysis, Diastolic BP before thrombolysis, glucose before thrombolysis, FBG before thrombolysis, Systolic BP at 2 hours after thrombolysis, Diastolic BP at 2 hours after thrombolysis, LDL after thrombolysis, TC after thrombolysis, Aspirin use within seven days after thrombolysis, clopidogrel use within seven days after thrombolysis, OCSP subtypes. Abbreviation: mRS, modified Rankin Scale; SICH, symptomatic intracranial hemorrhage; SITS-MOST, safe implementation of treatments in stroke-monitoring; ECASS II, second European–Australasian acute stroke study; NINDS, National Institute of Neurological Disorders and Stroke. Tables Abbreviation: TIA, transient ischemic attack; SBP, systolic blood pressure; DBP, diastolic blood pressure; WBC, white blood cell; PLT, platelet; INR, international normalized ratio; PT, prothrombin time; APTT, activated partial thromboplastin time; FBG, fibrinogen; NIHSS, National Institute of Health Stroke; LDL, low-density lipoprotein; TC, cholesterol; IQR, interquartile ranges; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large-artery atherosclerosis; Scale; SAO, small-artery occlusion; CE, cardioembolism. Table 2 Multivariate logistic regression analysis for risk factors of END able 2 Multivariate logistic regression analysis for risk factors of END Variables OR 95%CI P-value prior stroke/TIA 3.14 0.74-13.38 0.123 Glucose 1.10 1.03-1.18 0.004 FBG 1.07 0.91-1.26 0.426 LDL 1.24 0.79-1.95 0.361 TC 0.93 0.66-1.32 0.685 SBP on admission 1.00 0.98-1.02 0.748 DBP on admission 1.01 0.98-1.04 0.555 SBP at 2 hours after thrombolysis 1.02 1.00-1.04 0.107 DBP at 2 hours after thrombolysis 1.00 0.97-1.03 0.960 Taking aspirin within seven days before thrombolysis 0.25 0.14-0.44 <0.0001 Taking clopidogrel used within seven days before thrombolysis 0.39 0.19-0.82 0.013 bbreviation: OR, odds ratio; CI, confidence interval; TIA, transient ischemic attack; SBP, systolic blood ressure; DBP, diastolic blood pressure; FBG, fibrinogen; LDL, low-density lipoprotein; TC, cholesterol. Table 3 Outcomes after intravenous thrombolysis in END group versus non-END group Table 3 Outcomes after intravenous thrombolysis in END group versus non-END group Page 10/12 Page 10/12 Page 10/12 Outcomes No. Figure 2 Distribution of modified Rankin scale (mRS) score after intravenous Distribution of modified Rankin scale (mRS) score after intravenous thrombolysis in patients with acute ischemic stroke. thrombolysis in patients with acute ischemic stroke. Figures Page 11/12 Figure 1 Flow chart of eligible patients. END indicates early neurological deterioration; NIHSS, National Institute of Health Stroke; mRS, modified Rankin Scale; TIMS-China, Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China. Flow chart of eligible patients. END indicates early neurological deterioration; NIHSS, National Institute of Health Stroke; mRS, modified Rankin Scale; TIMS-China, Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China. Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. supplementaryNonstandardAbbreviationsandAcronyms0.docx This is a list of supplementary files associated with this preprint. Click to download. supplementaryNonstandardAbbreviationsandAcronyms0.docx Page 12/12	10,955
https://openalex.org/W4307011767	OpenAlex	Open Science	CC-By	2,022	THOUGHTS OF EASTERN THINKERS ABOUT MUSIC	Zarif R. Zoirov	Russian	Spoken	4,173	17,691	ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 ǩe_[kVS`SZSc _Shdfdda` UZV^rWUbcak^aX dbXi[S^o`n\`a_Xc ɮ ɹɮ ISSN 2181-9599 Doi Journal 10.26739/2181-9599 Doi Journal 10.26739/2181-9599 Джураева Нилуфар Далибаевна Джураева Нилуфар Далибаевна Муртазаева Рахбар Хамидовна тарих фанлари доктори, профессор, Ўзбекистон Миллий университети Муртазаева Рахбар Хамидовна тарих фанлари доктори, профессор, Ўзбекистон Миллий университети ур уф р тарих фанлари номзоди, доцент, Ўзбекистон жаҳон тиллари университети тарих фанлари номзоди, доцент, Ўзбекистон жаҳон тиллари университети 1 ǩe_[kVS`SZSc _Shdfdda` UZV^rWUbcak^aX dbXi[S^o`n\`a_Xc ɮ ɹɮ TОШКЕНТ-2022 TОШКЕНТ-2022 WUbcak^aX¬ ɮ ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 (2022) DOI http://dx.doi.org/10.26739/2181-9599-2022-SI-2 Бош муҳаррир ўринбосари: Заместитель главного редактора: Deputy Chief Editor: Бош муҳаррир ўринбосари: Заместитель главного редактора: Deputy Chief Editor: eSȗc[c[\_Sd^SȗSe]X`VSk[¬cXWS]i[a``n\daUXe¬ɖɮɸɮɖ Ковалев Борис Николаевич тарих фанлари доктори, профессор Санкт-Петербург Тарих институти Кобзева Ольга Петровна тарих фанлари доктори, профессор, Ўзбекистон Миллий университети Теймураз Ахалмосулишвили тарих фанлари номзоди, профессор, Телави давлат университети Ауанасова Алима Мусировна тарих фанлари доктори, профессор, Қозоғистон давлат тарихи институти Кебадзе Мадонна тарих фанлари доктори, Телави давлат университети Халикова Раҳбар Эргашевна тарих фанлари доктори, профессор, Тошкент давлат техника университети Бегалинова Калимаш Капсамаровна фалсафа фанлари доктори, профессор, Қозоғистон Миллий университети Аширов Адҳам Азимбаевич тарих фанлари доктори, профессор, ЎзР ФА Тарих институти Ишанходжаева Замира Райимовна тарих фанлари доктори, профессор, Ўзбекистон Миллий университети Хайдаров Муродилла Махмуталиевич тарих фанлари доктори, профессор, Ўзбекистон Миллий университети Агзамова Гулчеҳра Азизовна тарих фанлари доктори, профессор, ЎзР ФА Тарих институти Эшов Боходир Жўраевич тарих фанлари доктори, профессор Қарши давлат университети Алиева Лале тарих фанлари доктори, Озарбайжон давлат университети Бурдиашвили Майя тарих фанлари доктори, Телави давлат университети Маҳкамова Надира Рахмановна тарих фанлари доктори, профессор, Тошкент ахборот технологиялари университети Саипова Камола Давляталиевна тарих фанлари доктори, доцент, Ўзбекистон Миллий университети Кожакеева Ляззат Темировна тарих фанлари доктори, доцент, Хотин-қизлар давлат педагогика университети Абдуллаева Яхшибека Атамуратовна тарих фанлари доктори, доцент, Нукус давлат педагогика институти Исмаилова Алмаз тарих фанлари доктори, Доғистон мустақил университети Ульжаева Шохистахон Мамажоновна тарих фанлари доктори, Тошкент кимё - технология институти Дорошенко Татьяна Ивановна тарих фанлари номзоди, доцент, Ўзбекистон Миллий университети Бабаджанова Нодира Абдуллаевна тарих фанлари номзоди, доцент, Ўзбекистон давлат жаҳон тиллари университети Ширванова Тарана Амирага кызы тарих фанлари номзоди, доцент, Азербайджон давлат иқтисодиёт университети Мустафаева Нодира Абдуллаевна тарих фанлари доктори Ўзбекистон Фанлар академияси Иноятова Диларам Маниглиевна Тарих фанлари доктори (DSc) Ўзбекистон Миллий университети Ерметов Аваз Абдуллаевич тарих фанлари доктори, профессор, Ўзбекистон Миллий университети Одилов Аброр Анварович тарих фанлари номзоди, доцент, Ўзбекистон Миллий университети Джоробекова Айнур Эшимбековна тарих фанлари номзоди, профессор, Қозоғистон дипломатия академияси Page Maker \| Верстка \| Саҳифаловчи: Хуршид Мирзахмедов Контакт редакций журналов. www.tadqiqot.uz ООО Tadqiqot город Ташкент, улица Амира Темура пр.1, дом-2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Тел: (+998-94) 404-0000 Editorial staff of the journals of www.tadqiqot.uz Tadqiqot LLC the city of Tashkent, Amir Temur Street pr.1, House 2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Phone: (+998-94) 404-0000 2 For citation: Zarif R. Zoirov, THOUGHTS OF EASTERN THINKERS ABOUT MUSIC. Look to the past. 2022, Special issue 2, pp.61-67 For citation: Zarif R. Zoirov, THOUGHTS OF EASTERN THINKERS ABOUT MUSIC. Look to the past. 2022, Special issue 2, pp.61-67 АННОТАЦИЯ Ц Шарқ мусиқашунослик илмининг шаклланишидаги энг ахамиятли даврлардан бири Шарқ уйғониш даври ҳисобланади. Бу давр Х-ХI асрларга тўғри келади. Ўрганилаётган даврда шеърият ва мусиқа назарий ҳамда амалий жиҳатдан бир-бирига яқинлашди ва уйғунлашиб кетди. Ушбу ҳолат кейинги асрларда яшаб ижод этган шоирлар, мусиқачилар ижодида яққол акс этди. Шарқ алломалари томонидан яратилган асарларда мустақил мусиқашунослик фани асослари, умумий мусиқашуносликка оид тушунчалар ишлаб чиқилди, атамалар тизими яратилди. Калит сўзлар: мусиқа, Ибн Сино, Форобий, ал-Киндий, Беруний, Абу Абдуллоҳ Хоразмий. ШАРҚ МУТАФАККИРЛАРИНИНГ МУСИҚА ТЎҒРИСИДАГИ ФИКРЛАРИ ШАРҚ МУТАФАККИРЛАРИНИНГ МУСИҚА ТЎҒРИСИДАГИ ФИКРЛАРИ ɮ _f`WSc[YS¬daWXcYS`[X¬ɹ 1. Фирюза Абдурашидовна Мухитдинова ИНСОН ҚАДРИ ВА ШАЪНИНИ УЛУҒЛАГАН СИЁСИЙ ВА ҲУҚУҚИЙ ТАЪЛИМОТЛАРНИНГ ХУСУСИЯТЛАРИ……………………………………………………...5 2. Насиба Ибодуллаевна Абдуллаева ОЛИЙ ТАЪЛИМДАГИ БАДИИЙ ТАЪЛИМНИНГ РИВОЖИДА “ДАСТГОҲЛИ РАНГТАСВИР” КАФЕДРАСИНИНГ АҲАМИЯТИ ВА РОЛИ……………………………….19 3. Абдурахмон Абдумўминович Абдухалимов ВОДИЙ ВИЛОЯТЛАРИ МАКТАБЛАРИДА МИЛЛИЙ ТАЪЛИМ ТИЗИМИНИНГ ШАКЛЛАНИШ ВА РИВОЖЛАНИШ МАСАЛАЛАРИ………………………………………..30 4. Шаҳноза Асатуллаевна Абдураҳимова ЎЗБЕКИСТОН ССРДА ОЛИЙ МАЪЛУМОТЛИ ТИББИЁТ КАДРЛАРНИ ТАЙЁРЛАШ ҲОЛАТИ (1941-1991 ЙИЛЛАР)………………………………………………….38 5. Очил Бўриев, Шерзод Искандаров АМИР ТЕМУР ВА ТЕМУРИЙЛАР: САЙЙИДЛАР ҲОМИЙСИ……………………………...47 6. Dildora Doʼstmurodova XIX-XX ASRLARGA OID FARFOR KOLLEKSIYALARINI TADQIQ ETISH (TOSHKENT MUZEYLARI FAOLIYATI ASOSIDA)…………………………………………...54 7. Зариф Рашитович Зоиров ШАРҚ МУТАФАККИРЛАРИНИНГ МУСИҚА ТЎҒРИСИДАГИ ФИКРЛАРИ……………..61 8. Шерзод Абдиғаниевич Искандаров ЎЗБЕКИСТОН АРАБЛАРИНИНГ ЭТНОМАДАНИЙ ВА ДИНИЙ ЎЗЛИКНИ АНГЛАШИ……………………………………………………………..68 9. Шерзод Абдуғаниевич Искандаров ЎЗБЕКИСТОН АРАБЛАРНИНГ МОТАМ МАРОСИМ БИЛАН БОҒЛИҚ ТАОМЛАР……..75 10. Гулрух Акмал қизи Касимова АРХИВ ҲУЖЖАТЛАРИДА ТИББИЁТ ХОДИМЛАРИНИНГ ҒАЛАБАГА ҚЎШГАН ҲИССАСИНИ ЁРИТИЛИШИ…………………………………………81 11. Улуғбек Нигматов ТАСВИРИЙ САНЪАТДА КОМПОЗИЦИЯ……………………………………………………..87 12. Маҳфуза Бахриддиновна Маматова ШИМОЛИЙ ЧОЙ ЙЎЛИ ВА УНИНГ ТАРИХИНИ ЎРГАНИЛИШИ………………………..92 13. Dilmurod Sunatovich Omonov SHULLUKTEPA ARXEOLOGIK YODGORLIGIDA 1989-YILGI OLIB BORILGAN ARXEOLOGIK QAZISHMA ISHLARI…………………………………………….97 14. Абдилатип Абдирахимович Саримсоков ЎЗБЕК ТАҚВИМИЙ МАРОСИМЛАРИ: ЧИЛЛА 3 ¬ W ¬ 15. Баҳодир Босимович Тўйчибоев ПЕТРОГЛИФЛАРНИ ҲУЖЖАТЛАШТИРИШДА РАҚАМЛИ ТЕХНОЛОГИЯЛАРНИНГ ЎРНИ………………………………………………………………109 16. Феруза Раҳмоналиевна Ҳасанова ЗИЁВИДДИН ҲАЗИНИЙ МЕРОСИ ВА УНИ МУЗЕЙЛАШТИРИШ МАСАЛАЛАРИ…………………………………………………………………………………..115 17. Холлиев А.Г. РОССИЯ ИҚТИСОДИЁТИДА БРИТАНИЯ САРМОЯСИ (XIX АСРНИНГ ОХИРГИ ЧОРАГИ – ХХ АСР БОШИ)……………………………………...121 18. Бахтиёр Халмуратов, Абдуллажон Пазилов ФАРҒОНА ВОДИЙСИ ЎЗБЕКЛАРИДА ШОМОНЛИК ЭЪТИҚОДИ: ТАРИХИЙ ЎЗГАРИШЛАР КОНТЕКСТИДА…………………………………………………131 19. Азамат Холмуратович Шаимкулов ҚАЙТА ҚУРИШ ДАВРИДА ОММАВИЙ АХБОРОТ ВОСИТАЛАРИ ВА ФУҚАРОЛАР МУРОЖААТИ……………………………………………………………...137 20. Назиржон Салижонович Яқубов ЎЗБEКИСТОН РEСПУБЛИКАСИ КОНСТИТУЦИЯЛАРИ ВА САЙЛОВ ҲУҚУҚИ ТАРИХИДАН (Туркистон АССР, Хоразм ХСР, Бухоро ХСР ва Ўзбекистон ССР конституциялари мисолида)……………………………….145 21. Сулайманова Сохиба Баходировна ИЛК ЎРТА АСРЛАР ЁЗМА МАНБАЛАРИДА СУҒД ВОҲАСИ ТАРИХИ………………...155 4 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 Зариф Рашитович Зоиров АННОТАЦИЯ Ц Одним из важнейших периодов в становлении науки восточного музыковедения является Восточное Возрождение. Этот период соответствует X-XI векам. В изучаемый период поэзия и музыка сблизились и согласовались теоретически и практически. Эта ситуация ярко отразилась в творчестве поэтов и музыкантов, живших и творивших в последующие века. В трудах, созданных учеными Востока, были разработаны основы самостоятельной науки музыковедения, понятия, относящиеся к общему музыкознанию, создана система терминов. Ключевые слова: музыка, Ибн Сина, Фараби, аль-Кинди, Беруни, Абу Абдулла Хорезми. чевые слова: музыка, Ибн Сина, Фараби, аль-Кинди, Беруни, Абу Абдулла 61 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 Zarif R. Zoirov, Senior teacher of the Department of «Music Education» of Termiz State University ABSTRACT One of the most important periods in the formation of the science of Eastern musicology is the Eastern Renaissance. This period corresponds to the X-XI centuries. In the period under study, poetry and music have become closer and harmonized theoretically and practically. This situation was clearly reflected in the works of poets and musicians who lived and created in the following centuries. In the works created by the scholars of the East, the foundations of the independent science of musicology, concepts related to general musicology were developed, and a system of terms was created. Index Terms: music, Ibn Sina, Farabi, al-Kindi, Beruni, Abu Abdullah Khorezmi 1. Долзарблиги: Бугунги кунда миллий мусиқамиз тарихини ўрганишга бўлган қизиқиш тобора ортиб бормоқда. Айниқса, Форобий, Абу Али ибн Сино, Абу Райҳон Беруний, Абу Абдуллоҳ Хоразмий, ал-Киндий сингари буюк Шарқ мутафаккирларининг мусиқа бўйича билдирган фикрлари тўғрисида илмий жамоатчилик, оммавий ахборот воситалари ва ижтимоий тармоқларда турли-туман маълумотлар келтирилмоқда. Шунга қарамай, ушбу мавзуни тўла ўрганилган деб бўлмайди. Чунки, сўнгги йилларда ўтказилган тадқиқотлар илм-фан учун маълум бўлмаган кўпгина янгиликларни очиб бердики, улар илмий жамоатчиликда миллий мусиқамиз тарихига бўлган қизиқишнинг янада ортишига сабаб бўлмоқда. Бу эса, ушбу мавзунинг долзарб вазифага айланганлигидан далолат беради. 2. Методлар ва ўрганилганлик даражаси: Ушбу мақола тарихийлик ва илмийлик каби тарихий методлар асосида ўрганилган бўлиб, унда Шарқ мутафаккирларининг мусиқа тўғрисидаги фикрлари тўғрисидаги маълумотлар берилади. у р р Ўзбек миллий мусиқаси тарихи ва Шарқ мутафаккирларининг мусиқа тўғрисидаги фикрлари бўйича тадқиқотлар юртимизда ўтган асрдаёқ бошланган эди. Улардан атоқли олим А.Фитрат, рус олими В.Успенский, шунингдек, М.Хайруллаев, Т.Саломонова, А. Аминов, И.Ражабов, З.Эгамбердиев ва З.Эгамбедиева каби олимлар бир қатор тадқиқот ишларини олиб бордилар. Мустақиллик йилларида яратилган тадқиқотлар орасида Л.Бобоева [1], М.Қирғизов [2], С.Турсунов [3], И.Умаров [4], Г.Аҳмедова [5] сингари олимларнинг ишларига алоҳида тўхталиш лозим. Айтиш жоизки, уларнинг ишлари холисона руҳда ёзилган бўлиб, бугунги кун талабларига тўла жавоб беради. Шу билан бирга, янгидан-янги маълумотларнинг аниқланиши, илгари эълон қилинмаган материалларининг чоп этилиши бу борада бўйича жиддий изланишлар олиб борилишини талаб қилади. 3. Тадқиқот натижалари: 3. Тадқиқот натижалари: 3. Тадқиқот натижалари: Форобий қомусий олим бўлиши билан бирга буюк мусиқашунос ҳам эди. Форобий мусиқасининг аксарият ҳолларда инсоннинг нозик сезгилари, туғма истеъдод ва салоҳиятларининг меваси эканлигини таъкидлаш билан баробар, унинг, ўз навбатида, инсон маънавияти ва маърифатини, онги ва тушунчаларини тўлдирувчи руҳий неъмат эканлигини ўқтиради. “Исъҳоқ ибн Иброҳим ал-Мавсимий айтганки, «куйлар эркаклар тўқиган ва аёллар безаган гиламдир. Ушбу ҳайъатдаги маърифат ҳам ҳис билан сезилишга яқин бўлган ҳолдаги вужуд маърифати ёки уни сезги орқали топиш мумкин бўлган мазмун маърифатидир» [6] . Форобий куйларнинг турлари ва уларнинг инсон руҳиятига таъсири ҳақида гапириб, уларни уч турга ажратади: кўнгилга лаззат ва роҳат бағишловчи; бошқаси биринчи тур куйлар билан қўшилиб, кўнгилни таҳайюл билан бойитувчи; учинчи хил куйлар эса ҳайвонларнинг лаззати ва азияти билан боғлиқ инфиолот ва ахвол ҳақидаги мазмундан таркиб топади. 62 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 Форобий қарашларига кўра, барча табиий ва сунъий товушлар, ҳиссиёт ва кечинмалар моҳияти, мазмуни инсон камолотига, идрок қувватига боғлиқ бўлади. Форобий созанда, бастакор, мусиқа асбоблари устаси сифатида мусиқа тарихида ҳам ўз ўрнига эга. Айниқса, унинг «Китоб ал-мусиқий ал-Кабир» – «Катта мусиқа китоби» олим номини мусиқа назариётчиси ва амалиётчиси сифатида нафақат Шарққа, балки Ғарбга ҳам маълум қилди. Аввало, Форобий фанлар таснифида мусиқани математика илми таркибига киритади. Форобий «Ихсо ал-улум» асарида эса мусиқа илмини қуйидагича таърифлаб: «Мусиқа илмига келганда қисқача шуни айтиш мумкинки, у оҳангларнинг турларини, уларнинг тузилиши нималардан иборат эканлиги ва қайси йўллар билан ташкил топиши ҳамда қандай ҳолларда ўткир ва таъсирчан бўлишини ўрганади», -дейди ва мусиқа илмини амалий ҳамда назарий қисмларга ажратади [7]. «Китоб ал-мусиқий ал-Кабир» асарининг биринчи китобида мусиқий билимларнинг усул-қоидалари ва унга боғлиқ бўлган турли нарсаларни ўрганиш; иккинчи китобда замонавий мусиқа асбоблари ва улардан фойдаланиш йўллари, учинчисида эса, товушларнинг турлари ҳақидаги фикр баён этилади. Олим мусиқа илмини инсон билимлари тизимида тутган ўрни, бошқа илмлар билан алоқаси, мусиқий товушларнинг вужудга келиши ва ривожланишини таҳлил этар экан, мусиқа санъатининг ахлоқий эстетик жиҳатлари ҳақида ҳам ўз қарашларини баён этади. Ў Масалан: «Ўзбек мусиқаси тарихи» ўқув қўлланмасида Форобийнинг товушларнинг ҳарактерига кўра мусиқа асбоблари хилма-хил роль ўйнаганлиги ҳақидаги фикри келтирилади: «Жангларга мўлжалланган чолғу асбоблари мавжуд, уларнинг овози баланд ва кескин. Зиёфат ва рақслар учун, тўй ва қувноқ йиғилишлар учун, муҳаббат қўшиқлари учун ҳам махсус чолғу асбоблари бор. Айримларининг товуши кескин ва хазин бўлади; бир сўз билан айтганда улар шунчалик хилма-хилки, ҳаммасини санаб ўтиш қийин» [8]. Форобий мусиқа санъатини инсоннинг ахлоқий-эстетик такомили билан бирга, илмий билимлар тизимида ҳам катта аҳамиятга эга эканлигини кўрсатиб берди. 3. Тадқиқот натижалари: Бу борада М.Хайруллаев Форобийнинг мусиқа соҳасидаги тадқиқотчилар олдига қўйган талабларини келтирадики, мазкур талабларни бажариш ҳар бир илм одами учун зарур саналади: р ур р р р р у у рур «Назарий билимлардан хабардор бўлган етук санъаткор қайси билимда бўлмасин, шу уч нарсага эга бўлиши керак: у р ў р - Ҳар бир билимнинг усул ва қоидаларини тўла билиши лозим. - Ҳар бир билимнинг усул ва қоидаларини тўла билиши лозим. - Шу билимнинг турли ҳолатларида ҳосил бўлган усул ва қоидаларнинг натижаларини шарҳ ва изоҳ этишга қобил ва истеъдодли бўлиши керак. - Шу билимнинг турли ҳолатларида ҳосил бўлган усул ва қоидаларнинг натижаларини шарҳ ва изоҳ этишга қобил ва истеъдодли бўлиши керак. - Мусиқа санъати ҳақида нотўғри назариятга эга бўлган ёзувчиларнинг нотўғри назарияларини танқид эта билсин, хато ва тўғри назариялар ўртасидаги фарқни кўрсата билсин»[9]. - Мусиқа санъати ҳақида нотўғри назариятга эга бўлган ёзувчиларнинг нотўғри назарияларини танқид эта билсин, хато ва тўғри назариялар ўртасидаги фарқни кўрсата билсин»[9]. Кўриниб турибдики, Форобийнинг «Китоб ал-мусиқий ал-Кабир» асари ҳам назарий, ҳам ахлоқий-эстетик жиҳатдан қимматга эга бўлиб, инсоннинг ҳам ақлий, ҳам нафосат тарбиясида катта рол ўйнайди. Оҳанг ё бир неча товушларни белгили ва ажратилган ҳолда кетма-кет баён этиш ёки бир неча оҳангларни тартибли равишда зеҳнга таъсир этадиган ва турли таъсир кучига эга куйларни маълум ғояга хизмат килувчи сўзларда, истеъмол бўладиган қилиб бирин-кетин баён этишдир. Чунончи, инсон бир турли садо-оҳанг воситаси билан ўз фикрини баён қилиб, дилидаги хис-туйғуни ва мулохазасини бошқаларга етказади. Кунлардан бир куни Форобий Соҳиб Оббод шарафига берилган базмга кириб борибди. Хеч ким уни танимас экан. Форобий базмга кириб, бир чеккада жим ўтирибди. Чамаси, бирор соат ўтгандан сўнг мусиқа асбобини қўлига олиб, ажойиб бир куй чала бошлабди. Куйнинг биринчи қисмини чалганда атрофдагиларнинг ҳаммаси ниҳоятда хурсанд, кайфиятлари чоғ бўлибди. Куйнинг иккинчи қисмини эшитиб, ниҳоятда маъюс ҳолатга тушибдилар, йиғлаб кўз ёши тўка бошлабдилар. Куйнинг учинчи қисми ижро этилганда, ҳаммалари хаяжонга тушибдилар, сўнгра эса ухлаб қолибдилар. 63 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 Шунда меҳмон чолғу асбобининг дастасига: «Форобий келди, шу ерда бўлди-ю, кетди»,-деб ёзиб кетибди. Шунда меҳмон чолғу асбобининг дастасига: «Форобий келди, шу ерда бўлди-ю, кетди»,-деб ёзиб кетибди. Шунда меҳмон чолғу асбобининг дастасига: «Форобий келди, шу ерда бўлди-ю, кетди» -деб ёзиб кетибди Абу Али ибн Сино учун мусиқа ҳам инсон маънавиятини шакллантирувчи, ҳам бу маънавиятнинг ахлоқий-эстетик мезони ҳисобланган. У ўзининг «Аш-Шифо» асари муқаддимасида инсон маънавиятини ифодалашда товуш ва инсон сезгиларининг маданийлашуви ҳақида мулоҳазалар юритади. 3. Тадқиқот натижалари: У мусиқа пайдо бўлишининг асосий сабабларидан бири одамлар орасидаги маънавий эҳтиёж эканлигини, инсон мусиқа орқали ўз кайфияти, руҳиятида содир бўлаётган ходисалардан бошқаларни огоҳ этишини, мусиқа шу туфайли одамларнинг маданий ҳаёти ўзаро «ўртоқ»лик, боғлиқлик ҳаётининг ифодаси бўлишини кўрсатади. Ибн Сино мусиқа санъати одамларнинг маънавий камолотида катта ахамиятга эга эканлигини таъкидлайди. У товуш ва нутқни инсон камолотининг рамзи сифатида тушунади. Мусиқани кишиларга руҳий озуқа берувчи, жаҳолат ва нодонликдан қутулишда нажот бўлишини кўзда тутган Ибн Сино ўз асарларини «Аш-Шифо», «Ан-Нажот», «Донишнома» деб аташи бежиз эмас. Ибн Сино мусиқага доир рисолаларини алоҳида рисолаларидан ташқари бошқа соҳаларга, айниқса, тиббиёт илмига бағишланган асарларида ҳам баён этган. У «Тиб қонунлари»да қуйидагиларни ёзади: «...боланинг мижозини кучайтирмоқ учун, унга икки нарсани қўлламоқ керак. Бири, болани аста-секин тебратиш, иккинчиси, уни ухлатиш учун айтиш одат бўлиб қолган мусиқа ва аллалашдир. Шу иккисини қабул қилиш миқдорига қараб боланинг танаси билан бадантарбияга ва руҳи билан мусиқага бўлган истеъдоди ҳосил қилинади». Демак, мусиқа одамни нафақат жисмоний жихатдан, балки маънавий баркамол қилиш учун хизмат қилади. Ибн Сино замондоши, суриялик шоир ва мутафаккир ал-Маарининг мусиқаси ва унинг инсон руҳиятига таъсир қилиниши ҳақидаги фикрлари унинг «Рисолат-ул-гуфрон» асаридаги ривоятларда ўз ифодасини топган. Унинг фикрича ашула, куй «дардни қочиради, уйқуни ва ланжликни йўқ қилади». Ибн Сино мусиқани назарий ва амалий жиҳатдан Арасту даражасида билганлиги натижасида ўзининг «Рисолаи мусиқа» асарини ёзади. У ўрта аср мусиқачилигига ўзининг улкан ҳиссасини қўшди. Олимнинг мусиқа соҳасидаги қарашлари унинг ажойиб, мукаммал асари «Китоб ун- Нажот»да атрофлича, янада чуқурроқ даражада ривожлантирган. Ибн Сино мусиқа фанига атаб ёзган асари ҳақида шундай дейди: «Бу ёзган асаримда аждодларимиз хикматларидан беш нуқтаси жамланган бўлиб, беш нуқтанинг тўртинчиси «Мусиқа илми»дир». Олим мазкур асарда овозларнинг ёқимли ва ёқимсизлиги сабабларини очишга ҳаракат қилган. Абу Али ибн Синонинг мусиқа тарихига бағишланган асарлари ХI-ХII асрлардаёқ Европа халқлари тилига ўгирилиб, мактабларда дарслик сифатида ўқитилган. Ибн Синонинг дурдона асарлари ичида мусиқа фанига бағишланган илмий кузатишлари ва умумлашмалари ўзининг қадимийлиги, турли тадқиқотларга асосланганлиги билан умрбоқийдир [10]. Мусиқанинг инсон маънавиятида тутган ўрнига алоҳида ахамият берган Шарқнинг ахли донишлари куйнинг таълимий, тарбиявий таъсир кучига таянган ҳолда, аввало, хофиз ва созанданинг хулқ-атвори диёнат, эзгулик ҳамда нафосат мезонига айланмоғи керак деб қараганлар. Мусиқанинг инсон маънавиятига таъсири жуда қадимдан Шарқ мутафаккирлари ва мусиқашунослари фикрини банд қилган. «Араблар файласуфи» ал-Киндий мусиқани коинот гармонияси, инсон руҳияти гармониясига қиёсан ўрганади, инсонни жисмоний ва маънавий соғлом қилувчи омил сифатида идрок этади. У мусиқага доир рисолаларида куй, ранг ва ҳидларни эстетик идрок этиш жараёнидаги муштаракликни аниқлашга ҳам аҳамият беради. 3. Тадқиқот натижалари: Ал-Киндий Шарқ олимлари орасида биринчилардан бўлиб мусиқа назарияси ҳақида 64 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 рисолалар ёзибгина қолмай, мусиқанинг бадиий-эстетик ва тарбиявий жихатларига ҳам катта эътибор берди. рисолалар ёзибгина қолмай, мусиқанинг бадиий-эстетик ва тарбиявий жихатларига ҳам катта эътибор берди. рисолалар ёзибгина қолмай, мусиқанинг бадиий-эстетик ва тарбиявий жихатларига ҳам катта б б р р Ал-Киндий Шарқнинг буюк сиймолари-Хоразмий, Форобий, Беруний, Ибн Сино билан ёнма-ён қўйса бўладиган мутафаккир, араб мусиқасининг отаси ва энг етук файласуфидир. Ал-Киндий ёшлигидаёқ илмга берилди, ўз даврининг кўзга кўринган олимлари билан танишди, уларнинг асарларини мутолаа қилди, бир неча чет тилларни ҳам ўрганди. Аристотелнинг «Метафизика», «Категория» асарларини, Птолемейнинг «География», Эвклиднинг «Унсурлар» ва бошқа асарларни юнон тилидан арабчага ағдарди, баъзи бирларига шарҳлар ҳам ёзди. Ўрта Осиё, умуман, Шарқнинг дунёга машҳур алломалари каби ал-Киндий ҳам замонасининг барча фанларидан хабардор шахс бўлганлиги учун уни «Абул-ҳукамо» - ҳакимлар отаси деб аташган. Абу Райҳон Беруний ўзининг машҳур «Минералогия» асарида шу соҳага оид китоблар орасида ал-Киндийнинг рисоласидан бўлак дурустроқ асар кўрмаганлигини ёзади. Чиндан ҳам, ал-Киндий математика, астрономия, кимё, геология, геодезия, тарих каби фанлар билан теран шуғулланган. У араб олимлари орасида биринчи бўлиб материя, шакл, руҳ, вақт, макон ва замон каби фалсафий терминларни таърифлаб берди. Ал-Киндийнинг тиб илмида қилган катта хизмати шундаки, у мусиқа билан табобатнинг ўзаро алоқадорлигини аниқлаган. У мусиқанинг инсонга руҳий таъсири устида кўп шуғулланган ва бу соҳада анчагина мувафаққиятларга эришган. Ибн Сино руҳий касалликларни даволашда ал-Киндийнинг мусиқа билан даволаш соҳасидаги ишларига катта эътибор берган. Қомусий аллома ал-Киндийнинг ёзишича, куйлар билан гуллар ўртасида қандайдир ўхшашлик бор. Гуллардан тараладиган хушбўй киши руҳини кўтаради, кучига-куч қўшади. Мутафаккирнинг мусиқа ва табобатга доир ана шу мулоҳазалари ҳозирги кунда ҳам ўз қимматини йўқотмаган. Тадқиқотчилар Ибн Сино ҳам мусиқага оид асарлар яратиб, уларнинг фақат бир қисми бизгача етиб келган деб маълумот берадилар. Шулардан бири «Мусиқа билимига оид тўплам» бўлиб, бунда товушнинг сезги аъзоларига таъсири, унинг ёқимли ва ёқимсизлиги, товушни эшитганда лаззатланиш ё нафратланиш хиссининг пайдо бўлиши каби масалаларга тўхтайди. Унда мусиқанинг киши ҳаётигача қанчалик зарурлиги ҳақида ҳам фикрлар баён этилади. Олимнинг фикрича, инсон табиатан ёқимли нарсалар орқали енгил торца, унинг аксида ороми йўқолиб, нафрати пайдо бўлади. Ибн Сино мусиқа овозларининг киши руҳига таъсири ҳақида ноёб фикрлар баён этади [11]. Абу Абдуллоҳ Хоразмийнинг бизгача етиб келган ягона асари «Мафотих ал-Улум»дир. Араб тилида битилган Шарқдаги илк қомусий асарлардан бўлган «Илмлар калитлари» 976- 991 йиллар орасида яратилган бўлиб, унда ўз даврида маълум бўлган фаннинг барча соҳалари қамраб олинган ва уларнинг асослари баён этилган. 6. Абу Наср ал-Форобий. Китабул-мусиқа ал-Кабир. Арабчадан А.Назаров таржимаси. – Тошкент, 1997. –Б. 14-15. (Abu Nasr al-Farabi. Book of music al-Kabir. Translated from Arabic by A. Nazarov. –Tashkent, 1997. –P. 14-15.) 3. Тадқиқот натижалари: У жуда эътиборли ва ишонарли бир манба бўлганки, Ибн Сино ҳам ўзининг «Ақлий илмлар бўлимлари» рисоласида ундан иқтибослар келтиради. «Илмлар калитлари» икки қисмга-мақолага бўлинган, бу қисм – мақолалар таркибида 90 фаслни бирлаштирувчи 15 боб бор. Мусиқага бағишланган боб уч фаслдан иборат. Хоразмий мусиқага бағишланган боб таркиби ҳақида шундай ёзади: «Иккинчи қисмнинг еттинчи боби мусиқа ҳақида ва у уч фаслдан иборат. Биринчи фасл мусиқа асбобларининг аталишлари ва улар билан боғлиқ нарсалар ҳақида, иккинчи фасл донишмандлар китобларида келтирилган мусиқий жамлар ҳақида, учинчи фасл истеъмолдаги ийқолар - ритмлар ҳақида». Мусиқа асбобларига бағишланган биринчи фаслда ўн тўртта чолғу санаб ўтилиб, номлари келтирилади. Барбат - уднинг торлари, пардалари ва бошқа қисмлари баён этилади, созлаш изоҳланади. Бу фаслнинг эътиборли жойи шундаки, баъзи мусиқа асбобларига қисқа шарҳ ҳам, уларнинг қисмларининг аталишлари ҳам берилган. Масалан: Форобий, Ибн Сино ва Ибн Зайланинг мусиқашуносликка оид асарларида орган мусиқа асбобининг номигина келтирилиб, унинг шакли-шамоили берилмаган ҳолда, Абу Абдуллоҳ Хоразмийнинг асарида шундай дейилади: «Орган -юнонликлар ва римликларнинг мусиқа асбоби. Қора мол терисидан 65 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 №SI-2 \| 2022 ишланган, бир-бирига уланган катта уч чарм меш – босқон бўлиб, ўртадагисининг бошига катта меш – босқон ўрнатилади ва унга туйнукчалари маълум нисбатда жойлашган мис найчалар маҳкамланади ва улардан созанда истаги бўйича ёқимли, қувноқ ва хазин товушлар чиқаради». Демак, орган – ҳаво тўлдирилган босқонлардан бирига, яъни дам берувчисига – махсус мослама – босқонга босилганда, уларга ўрнатилган найлар товуш чиқарган. Алишер Навоий даврида ҳам орган кўп машҳур бўлган. р р ў ур ў Асарнинг жамларга бағишланган иккинчи фаслда товуш, нағма – тон, буъд – интервал, жинс – интерваллар тизими, тетрахорд ва жам – жинслар тизими, гамма – товушқаторлар ҳақида баҳслар олиб борилган. Ийқолар – ритмлар ҳақидаги учинчи фаслда қуйидаги саккиз ийқо санаб ўтилади: Хазаж. Хафиф ур-рамал. Рамал. Сақил ур-рамал. Ас-сақил ус-саний. Хафиф ус-сақил уссаний- Махурий. Ас-сақил ул-аввал. Хафиф ус-сақил ул-аввал. Абу Абдуллоҳ Хоразмийнинг мусиқа илмига оид қарашлари Форобий ва Ибн Синоникидек қамрови кенг эмас, аммо у айрим янги мусиқашунослик атамаларини қўллайди. Масалан: мусиқор – мусиқачи, бастакор атамасини, қадимий юнон торли тирнама мусиқа асбоблари қитора – гитара, лур – лира кабиларнинг аталишларини келтиради. У келтирган лур – лиранинг умумий шакли ҳозирда мусиқа эмблемаси сифатида қабул қилинган, мусиқор - мусиқачи, бастакор атамаси эса кўп шарқ тилларида хануз истеъмолдадир. «Илмлар калитлари» китобининг мусиқага бағишланган қисми немис ва инглиз тилларига ўгирилган, танқидий матни нашр этилган, форс тилига ағдарилиб нашрдан чиққан, ўзбек тилида бу борада изланиш бор. Асарнинг қўлёзма матни дунёнинг кўпгина кутубхоналарида сақланмоқда. 3. Тадқиқот натижалари: у у р Абу Абдуллоҳ Хоразмийнинг «Илмлар калитлари» асарининг мусиқага оид бобини тадқиқ этиш ҳозирги замон миллий ўзбек мусиқасининг устивор асоси – мақомлар ва мақомлар масаласига доир муаммоларни тўғри ҳал қилишда ҳам қимматли манбадир. Абу Абдуллоҳ Хоразмийнинг «Илмлар калитлари» асарининг мусиқага оид бобини тадқиқ этиш ҳозирги замон миллий ўзбек мусиқасининг устивор асоси – мақомлар ва мақомлар масаласига доир муаммоларни тўғри ҳал қилишда ҳам қимматли манбадир. 5. Аҳмедова Г. Сурхондарё бахшилари // «Ўтмишга назар» журнали, 2020 йил, №SI-4. –54- 58 б. (Ahmedova G. Surkhandarya bakhshis// journal «Look to the past», 2020, №SI-4. –54-58 p.) 1. Бобоева Л. Муғом ва мусулмон маданияти //«Санъат» журнали, 1990 йил. №9. (Boboeva L. Mug’om and Muslim culture // journal «Sanat», 1990. №9.) 4. Хулосалар. Шундай қилиб, айнан IХ-ХI асрларда мусиқа фанига асос солиниб, бу юз йилликларда яшаган мутафаккирларнинг деярли барчаси бу фан билан шуғулланганлар. IХ асрдаёқ мусиқага бағишланган рисолалар пайдо бўлган. Хусусан, уларда мусиқанинг назарий асослари тадқиқ этилади. Мусиқа асбоблари илмий тасвирланади, таникди ижрочилар номи, уларнинг репертуари, кенг тарқалган мусиқа жанрларининг номлари келтирилади. Форобий, Ибн Сино ва бошқа алломаларнинг ўрта асрда илм, маданиятни юқори даражада акс эттирган асарлари ана шу давр мусиқа маданиятини ўрганишимизда ҳам мухим манба бўлиб хизмат қилади. Иқтибослар/ Сноски/References: Иқтибослар/ Сноски/References: 1. Бобоева Л. Муғом ва мусулмон маданияти //«Санъат» журнали, 1990 йил. №9. (Boboeva L. Mug’om and Muslim culture // journal «Sanat», 1990. №9.) g j ) 2. Қирғизов М. Оҳангда жилоланган қадриятлар. –Тошкент: Фан, 1995. (Kirghizov M. Values polished in tone. -Tashkent: Fan, 1995.) Ў 3. Турсунов С.Н., Пардаев Т.Р., Турсунова Н.М., Муртазоев Б. Ўзбекистонда бахшичилик санъатининг шаклланиши ва тараққиёти тарихи. –Тошкент, 2015. (Tursunov S.N., Pardaev T.R., Tursunova N.M., Murtazoev B. The history of the formation and development of the art of bakhshi in Uzbekistan. –Tashkent, 2015.) 4. Умаров И. Бадаш: тарих ва анъана. –Термиз: Сурхоннашр, 2018. (Umarov I. Badash: history and tradition. –Termiz: Surkhannashr, 2018) Ў 5. Аҳмедова Г. Сурхондарё бахшилари // «Ўтмишга назар» журнали, 2020 йил, №SI-4. –54- 58 б. (Ahmedova G. Surkhandarya bakhshis// journal «Look to the past», 2020, №SI-4. –54-58 p.) 5. Аҳмедова Г. Сурхондарё бахшилари // «Ўтмишга назар» журнали, 2020 йил, №SI-4. –54- 58 б. (Ahmedova G. Surkhandarya bakhshis// journal «Look to the past», 2020, №SI-4. –54-58 p.) 66 ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ ǩe_[kVS`SZSc¬UZV^rWUbcak^aX¬ ɮ №SI-2 \| 2022 №SI-2 \| 2022 7. Хайруллаев М. Форобий рисоласидан намуналар. // «Уйғониш даври ва Шарқ мутафаккири» китобига илова. -Тошкент: Ўзбекистон. 1971. –Б. 295. (Khairullaev M. Samples from Pharaobi's treatise. // Appendix to the book «Renaissance and Eastern Thinker». –Tashkent: Uzbekistan. 1971. –P. 295.) Ў 7. Хайруллаев М. Форобий рисоласидан намуналар. // «Уйғониш даври ва Шарқ мутафаккири» китобига илова. -Тошкент: Ўзбекистон. 1971. –Б. 295. (Khairullaev M. Samples from Pharaobi's treatise. // Appendix to the book «Renaissance and Eastern Thinker». –Tashkent: Uzbekistan. 1971. –P. 295.) 8. Ўзбек мусиқаси тарихи. Мусиқа олий ўқув юртларининг студентлари учун қўлланма. Тузувчи: Т.Е.Саломонова. -Тошкент: Ўқитувчи. 1981. –Б. 11. (History of Uzbek music. Guide for students of music universities. Compiled by: T.E. Salomonova. -Tashkent: O’qituvchi. 1981. –P. 11.) 8. Ўзбек мусиқаси тарихи. Мусиқа олий ўқув юртларининг студентлари учун қўлланма. Тузувчи: Т.Е.Саломонова. -Тошкент: Ўқитувчи. 1981. –Б. 11. (History of Uzbek music. Guide for students of music universities. Compiled by: T.E. Salomonova. -Tashkent: O’qituvchi. 1981. –P. 11.) q ) 9. Хайруллаев М. Форобий рисоласидан намуналар. – Б.156. (Khairullaev M. Samples from Pharaobi's treatise. –P. 156.) 9. Хайруллаев М. Форобий рисоласидан намуналар. – Б.156. (Khairullaev M. Samples from Pharaobi's treatise. –P. 156.) Ў 10. Аминов А. Алломанинг мусиқа мероси // «Ўқитувчи» газетаси, 1980 йил, 2 июль. (Aminov A. Alloma's musical heritage // «O’qituvchi» newspaper, July 2, 1980.) 10. Аминов А. Алломанинг мусиқа мероси // «Ўқитувчи» газетаси, 1980 йил, 2 июль. (Aminov A. Alloma's musical heritage // «O’qituvchi» newspaper, July 2, 1980.) в А. Алломанинг мусиқа мероси // «Ўқитувчи» газетаси, 1980 йил, 2 июль. v A. Alloma's musical heritage // «O’qituvchi» newspaper, July 2, 1980.) g q p p y ) в А. Абу Али ибн Сино. Ҳаёти ва ижодий мероси. -Тошкент: Фан, 1980. –Б. 137- 11. Ирисов А. Абу Али ибн Сино. Ҳаёти ва ижодий мероси. -Тошкент: Фан, 1980. –Б. 137- 139. (Irisov A. Abu Ali ibn Sina. Life and creative heritage. -Tashkent: Fan, 1980. –P. 137- 139.) 11. Ирисов А. Абу Али ибн Сино. Ҳаёти ва ижодий мероси. -Тошкент: Фан, 1980. –Б. 137- 139. (Irisov A. Abu Ali ibn Sina. Life and creative heritage. -Tashkent: Fan, 1980. –P. 137- 139.) 67 166 ISSN 2181-9599 Doi Journal 10.26739/2181-9599 ǩe_[kVS`SZSc _Shdfdda` UZV^rWUbcak^aX dbXi[S^o`n\`a_Xc ɮ ɹɮ Контакт редакций журналов. www.tadqiqot.uz ООО Tadqiqot город Ташкент, улица Амира Темура пр.1, дом-2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Тел: (+998-94) 404-0000 Editorial staff of the journals of www.tadqiqot.uz Tadqiqot LLC the city of Tashkent, Amir Temur Street pr.1, House 2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Phone: (+998-94) 404-0000 ISSN 2181-9599 Doi Journal 10.26739/2181-9599 Doi Journal 10.26739/2181-9599 166 Контакт редакций журналов. www.tadqiqot.uz ООО Tadqiqot город Ташкент, улица Амира Темура пр.1, дом-2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Тел: (+998-94) 404-0000 Editorial staff of the journals of www.tadqiqot.uz Tadqiqot LLC the city of Tashkent, Amir Temur Street pr.1, House 2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Phone: (+998-94) 404-0000 Контакт редакций журналов. www.tadqiqot.uz ООО Tadqiqot город Ташкент, улица Амира Темура пр.1, дом-2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Тел: (+998-94) 404-0000 Editorial staff of the journals of www.tadqiqot.uz Tadqiqot LLC the city of Tashkent, Amir Temur Street pr.1, House 2. Web: http://www.tadqiqot.uz/; Email: info@tadqiqot.uz Phone: (+998-94) 404-0000	1,969

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