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https://openalex.org/W2977754774	OpenAlex	Open Science	CC-By	2,019	EXPRESSION DENSITY OF RECEPTORS FOR IMMUNOREGULATORY MEDIATORS AS A MODULATORY COMPONENT OF BIOLOGICAL EFFECTS OF MEDIATORS UPON CELLS (PART 2)	С. В. Сенников	Russian	Spoken	11,460	31,818	Адрес для переписки: Сенников Сергей Витальевич ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии» 630099, Россия, г. Новосибирск, ул. Ядринцевская, 14. Тел.: 8 (383) 222-19-10. E mail: sennikovsv@gmail com Сенников Сергей Витальевич ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии» 630099, Россия, г. Новосибирск, ул. Ядринцевская, 14. Тел.: 8 (383) 222-19-10. E il ik @ il ПЛОТНОСТЬ ЭКСПРЕССИИ РЕЦЕПТОРОВ К ИММУНОРЕГУЛЯТОРНЫМ МЕДИАТОРАМ КАК МОДУЛИРУЮЩИЙ КОМПОНЕНТ БИОЛОГИЧЕСКИХ ЭФФЕКТОВ МЕДИАТОРОВ НА КЛЕТКУ (ЧАСТЬ 2) Сенников С.В.1, 3, Альшевская А.А.1, Жукова Ю.В.1, Беломестнова И.А.1, Караулов А.В.2, Лопатникова Ю.А.1 Резюме. В обзоре обобщены последние мировые научные данные о роли рецепторов к иммуно- медиаторам в регуляции биологических эффектов, оказываемых на клетки. Для основных классов регуляторов иммунной системы (интерлейкинов, интерферонов, факторов роста и факторов некро- за опухоли) представлены варианты участия рецепторов как компонентов взаимодействия цитокин/ клетка на примере in vitro и in vivo исследований. Показана способность показателей экспрессии рецепторов менять характер и тип данного взаимодействия. Проанализированы данные об участии рецепторов к регуляторным молекулам в развитии иммуноопосредованных заболеваний различного генеза. Продемонстрировано, что изменение уровня экспрессии рецепторов имеет большое значе- ние в оценке функционального ответа клетки на медиатор и в развитии патологических состояний. В исследованиях подтверждены данные о влиянии плотности рецепторов на процессы пролифе- рации и апоптоза, а также обменные процессы, что является триггером в развитии аутоиммунных, онкологических и дистрофических заболеваний. Для всех рассмотренных классов регуляторных молекул характерным является изменение плотности экспрессии рецепторов как одного из ключе- вых моментов регулирования функциональной активности клеток. Таким образом, изучение уровня экспрессии рецепторов на мембране клеток является важным в понимании патогенеза, а изменение уровня экспрессии может рассматриваться как терапевтическая мишень в лечении различных за- болеваний. Адрес для переписки: Сенников Сергей Витальевич ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии» 630099, Россия, г. Новосибирск, ул. Ядринцевская, 14. Тел.: 8 (383) 222-19-10. E-mail: sennikovsv@gmail.com Address for correspondence: Sennikov Sergey V. Research Institute of Fundamental and Clinical Immunology 630099, Russian Federation, Novosibirsk, Yadrintsevskaya str., 14. Phone: 7 (383) 222-19-10. E-mail: sennikovsv@gmail.com Образец цитирования: С.В. Сенников, А.А. Альшевская, Ю.В. Жукова, И.А. Беломестнова, А.В. Караулов, Ю.А. Лопатникова «Плотность экспрессии рецепторов к иммунорегуляторным медиаторам как модулирующий компонент биологических эффектов медиаторов на клетку» (часть 2) // Медицинская иммунология, 2019. Т. 21, № 3. С. 379-396. doi: 10.15789/1563-0625-2019-3-379-396 © Сенников С.В. и соавт., 2019 For citation: S.V. Sennikov, A.A. Alshevskaya, Yu.V. Zhukova, I.A. Belomestnova, A.V. Karaulov, Yu.A. Lopatnikova “Expression density of receptors for immunoregulatory mediators as a modulatory component of biological effects of mediators upon cells” (part 2), Medical Immunology (Russia)/ Meditsinskaya Immunologiya, 2019, Vol. 21, no. 3, pp. 379-396. doi: 10.15789/1563-0625-2019-3-379-396 DOI: 10.15789/1563-0625-2019-3-379-396 Ключевые слова: цитокины, рецепторы, иммунорегуляция, иммунопатология, иммунотерапия, факторы роста, интерлейкины, интерфероны, фактор некроза опухоли Ключевые слова: цитокины, рецепторы, иммунорегуляция, иммунопатология, иммунотерапия, факторы роста, интерлейкины, интерфероны, фактор некроза опухоли Address for correspondence: Sennikov Sergey V. Research Institute of Fundamental and Clinical Immunology 630099, Russian Federation, Novosibirsk, Yadrintsevskaya str., 14. Phone: 7 (383) 222-19-10. E-mail: sennikovsv@gmail.com Medical Immunology (Russia)/ Meditsinskaya Immunologiya 2019, Vol. 21, 3, pp. 379-396 © 2019, SPb RAACI Medical Immunology (Russia)/ Meditsinskaya Immunologiya 2019, Vol. 21, 3, pp. 379-396 © 2019, SPb RAACI Медицинская иммунология 2019, Т. 21, № 3, стр. 379-396 © 2019, СПб РО РААКИ Медицинская иммунология 2019, Т. 21, № 3, стр. 379-396 © 2019, СПб РО РААКИ Медицинская иммунология 2019, Т. 21, № 3, стр. 379-396 © 2019, СПб РО РААКИ Обзоры Reviews Обзоры Reviews Работа поддержана грантом Сеченовского университета 2018. Данная работа является продолжением обзора о рецепторах к иммунорегуляторам, который был опубликован ранее [1]. Хемокиновые рецепторы Sennikov S.V.a, c, Alshevskaya A.A.a, Zhukova Yu.V.a, Belomestnova I.A.a, Karaulov A.V.b, Lopatnikova Yu.A.a a Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation b First Moscow I. Sechenov State Medical University, Moscow, Russian Federation с Novosibirsk State University, Novosibirsk, Russian Federation Abstract. The present review article summarizes the latest world scientific data on the role of receptors for immune mediators in regulating biological effects on the cells. For the main classes of immune regulators (interleukins, interferons, growth factors and tumor necrosis factors), the variants are presented for participation of receptors as components of cytokine/cell interaction, as proven by in vitro and in vivo studies. Ability of the receptors expression to modify characteristics and type of these interactions is shown. The data on participation of receptors for regulatory molecules in development of immune-mediated diseases of various genesis have been analyzed. It was demonstrated that the changes in the receptor expression are of great importance when evaluating functional response of the cells to the mediators and in development of pathological conditions. Current studies confirmed the data suggesting effects of receptor density upon the processes of proliferation and apoptosis, as well as metabolic processes that trigger development of autoimmune, oncological and dystrophic diseases. For all the considered classes of regulatory molecules, the change in the density of receptor expression is one of the key aspects in regulating functional activity of the cells. Thus, studying expression levels of receptors on the cell membrane is important in understanding pathogenesis, whereas changing expression level may be considered as a therapeutic target in the treatment of various diseases. Keywords: cytokines, receptors, immunoregulation, immunopathology, immunotherapy, growth factors, interleukins, interferons, tumor necrosis factor вышению их подвижности за счет повышения синтеза внутриклеточного актина, его полимери- зации и перераспределения в клетке. В результате образуются псевдоподии, способствующие про- движению и инвазии опухолевых клеток. С по- мощью такого механизма метастазируют в лим- фоидные органы клетки меланомы кожи [61], цервикальной интраэпителиальной неоплазии (предраковое состояние плоскоклеточного рака шейки матки) [62], рака молочной железы [45], плоскоклеточного рака пищевода [16], желуд- ка [40]. Помимо влияния на морфологию опу- холевых клеток, хемокиновые рецепторы, экс- прессирующиеся на мембране раковых клеток и взаимодействующие со своими лигандами, способствуют повышению их функциональной активности. Клетки в ответ секретируют фак- торы роста, в частности VEGF [60], в результате чего происходит увеличение плотности лимфа- тических сосудов в опухоли и тем самым увели- чивается лимфогенное распространение рака. Общей тенденцией, обозначаемой авторами при исследовании изменения экспрессии хемокино- вых рецепторов на поверхности клеток клиниче- Работа поддержана грантом Сеченовского университета 2018. For citation: S.V. Sennikov, A.A. Alshevskaya, Yu.V. Zhukova, I.A. Belomestnova, A.V. Karaulov, Yu.A. Lopatnikova “Expression density of receptors for immunoregulatory mediators as a modulatory component of biological effects of mediators upon cells” (part 2), Medical Immunology (Russia)/ Meditsinskaya Immunologiya, 2019, Vol. 21, no. 3, pp. 379-396. С.В. Сенников, А.А. Альшевская, Ю.В. Жукова, И.А. Беломестнова, А.В. Караулов, Ю.А. Лопатникова «Плотность экспрессии рецепторов к иммунорегуляторным медиаторам как модулирующий компонент биологических эффектов медиаторов на клетку» (часть 2) // Медицинская иммунология, 2019. Т. 21, № 3. , С. 379-396. doi: 10.15789/1563-0625-2019-3-379-396 © Сенников С.В. и соавт., 2019 , С. 379-396. doi: 10.15789/1563-0625-2019-3-379-396 © Сенников С.В. и соавт., 2019 pp doi: 10.15789/1563-0625-2019-3-379-396 DOI: 10.15789/1563-0625-2019-3-379-396 379 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. et al. Сенников С.В. и др. Хемокиновые рецепторы Хемокиновые рецепторы вместе со своими лигандами цитокиновой природы – хемокина- ми – играют важную роль в жизни клеток и все- го организма: это хемотаксис и миграция кле- ток, определяющие развитие организма в целом и процессы, возникающие во время нарастания воспаления, влияя на подвижность, прикрепле- ние и инвазию лейкоцитов через стенку сосуда в паренхиму тканей [6]. Рецепторы хемокинов, как правило, экспрес- сируются на иммунных клетках, однако злока- чественные клетки при многих онкологических заболеваниях также экспрессируют некоторые рецепторы хемокинов, которые обычно не обна- руживают на их нормальных аналогах. Было за- мечено, что увеличение экспрессии некоторых хемокиновых рецепторов (CXCR7, CCR7, CCR8, CXCR4) на опухолевых клетках способствует по- 380 2019, Vol. 21, 3 2019, Т. 21, № 3 Рецепторы к иммунорегуляторам Receptors for immune regulators индуцируя апоптоз или повышая выживаемость и пролиферацию клеток. Между сигналами, ко- торые вызывают эти противоположные эффек- ты, существует баланс, позволяющий сохранять гомеостаз нормальных клеток. В настоящее вре- мя идентифицировано по меньшей мере около 26 рецепторов семейства TNF и около 18 лигандов семейства TNF. ских образцов, было то, что высокая экспрессия хемокиновых рецепторов коррелирует с тяже- стью онкологической патологии, увеличением риска развития рецидива, а также смерти. Па- циенты, чьи образцы имели высокую плотность этих рецепторов, имели более неблагоприятный прогноз заболевания, в отличие от пациентов, образцы ткани которых имели низкую экспрес- сию рецепторов либо не имели ее вообще. индуцируя апоптоз или повышая выживаемость и пролиферацию клеток. Между сигналами, ко- торые вызывают эти противоположные эффек- ты, существует баланс, позволяющий сохранять гомеостаз нормальных клеток. В настоящее вре- мя идентифицировано по меньшей мере около 26 рецепторов семейства TNF и около 18 лигандов семейства TNF. Надсемейство TNF-рецепторов можно раз- делить на две группы – содержащие и не содер- жащие домен смерти. В первую группу входят TNFR1 и FasR. Во вторую – TNFR2, CD40, CD30, DR4, DR5 и др. Все рецепторы широко распро- странены на клетках различных типов тканей и играют ключевые роли во многих физиологиче- ских процессах, включая развитие лимфоидной и нервной тканей, врожденный и приобретен- ный иммунитет, поддержание гомеостаза. Однако в отношении экспрессии CX3CR1 хемокинового рецептора на клетках колорек- тального рака были получены обратные резуль- таты [18]. Низкая экспрессия этого рецептора приводила к уменьшению общей выживаемости и увеличению частоты прогрессирования. Также в исследовании было отмечено, что сигнализа- ция через CX3CR1 приводит к достаточно силь- ному адгезивному взаимодействию между рако- выми клетками, предотвращая, таким образом, их распространение по организму. Известно два типа рецепторов, взаимодей- ствующих с TNFα. Хемокиновые рецепторы Взаимодействие цитокина с TNFR1 может обеспечивать, помимо воспали- тельных эффектов, выживаемость клетки и апоп- тоз. Связываясь со вторым типом рецептора, TNFα стимулирует пролиферацию, выживае- мость клеток и продукцию ими провоспалитель- ных медиаторов, в том числе цитокинов [8]. Важ- но, однако, учитывать не только тип рецептора, с которым связывается TNFα, но и плотность этого рецептора на поверхности клетки-мишени, так как даже внутрипопуляционный состав кле- ток может отличаться по уровню экспрессии ре- цепторов. Основные субпопуляции иммуноком- петентных клеток (Т-лимфоциты, В-лимфоциты и моноциты) отличаются по экспрессии ре- цепторов 1 и 2 типа как в норме [34], так и при патологии [58], при этом изменение процента клеток, экспрессирующих соответствующий ре- цептор в субпопуляции, не ассоциировано с из- менением плотности экспрессии рецепторов на данных клетках. При этом было установлено, что для различных заболеваний (в частности, для ревматоидного артрита) может быть харак- терно преобладание определенных вариантов аллельного полиморфизма рецепторов к TNFα, что также может иметь значение в развитии за- болевания [59]. Хемокиновые рецепторы не только опреде- ляют характер и тип развития онкологической патологии, но и играют роль в развитии ин- фекционных заболеваний. Было показано [24], что высокая экспрессия корецепторов CCR5 и CXCR4 способствует проникновению и ре- пликации макрофаготропного и Т-клеточно- тропного ВИЧ-1, соответственно, в CD4- позитивные клетки организма. В частности, были определены возможные механизмы сокращения уровня экспрессии CCR5 на лимфоцитах – ге- терозиготность по мутации Δ32 гена CCR5 [49], полиморфизм гена хемокиновых рецепторов CCR5-2459 A/G и CCR2-64I [26]. Высокая плотность экспрессии CX3CR1 в атеросклеротических бляшках сонных артерий приводила к развитию в них воспалительного процесса, что увеличивало риск разрыва и обра- зования тромбоза. При этом низкая экспрессия CX3CR1 характеризовалась наличием фиброза, как показателя стабильности бляшки. Эти дан- ные свидетельствуют о том, что оценка уровня экспрессии CX3CR1 может являться предикто- ром нестабильности атеросклеротической бляш- ки [41]. Таким образом, изменение плотности экс- прессии хемокиновых рецепторов на клетках может определять морфологические и функци- ональные изменения в клетках, что приводит к развитию патологического процесса. Hijdra D. и соавт. в своей работе показали, что три основные субпопуляции моноцитов крови отличаются по степени экспрессии рецепто- ров 1 и 2 типа к TNFα. Классические моноци- ты (CD14++CD16-) экспрессируют небольшое количество как TNFR1, так и TNFR2. Неклас- сические моноциты (CD14+CD16++) имеют вы- сокий уровень экспрессии 2 типа рецептора и низкий 1 типа. Промежуточные моноциты Рецепторы семейства TNF и рецепторы смерти (death receptors) Рецепторы семейства TNF и рецепторы смерти (death receptors) Члены семейства факторов некроза опухолей (TNF), включая лиганды и соответствующие ре- цепторы, регулируют рост нормальных клеток, 381 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. et al. Сенников С.В. и др. (CD14++CD16+) в большей степени экспрессиру- ют TNFR1, в отличие от двух других популяций, и занимают переходное по уровню экспрессии TNFR2. Такое распределение экспрессии ре- цепторов к TNFα среди моноцитов наблюдалось на клетках как здоровых доноров, так и пациен- тов с саркоидозом [25]. цепторов, в то время как преобладающая актив- ность TNFR1 может опосредовать уменьшение числа клеток и апоптоз, наблюдаемый в РК- дифференцированных нейронах. TNFα-индуцированное увеличение пролифе- рации было обнаружено при культивировании человеческих миофибробластов, несущих оба типа рецептора к TNFα, с цитокином в различ- ных дозах (0,1-10 нг/мл), с максимальным эф- фектом при 1 нг/мл [51]. Кроме того, культиви- рование миофибробластов в присутствии TNFα повышало инвазивную способность клеток и се- крецию ими матричной металлопротеиназы 9 (ММР-9), необходимой для процессов ремоде- лирования миокарда. В этом случае максималь- ный эффект наблюдался при 10 нг/мл цитокина в культуре. Однако TNFα вызывал увеличение пролиферативного ответа только в половине ис- следованных клеточных популяций, что, по мне- нию авторов, может быть связано с различным уровнем экспрессии обоих типов рецептора на клетках. Обнаруженные эффекты авторы свя- зывают с сигнализацией цитокина через 1 тип рецептора, поскольку его блокирование инги- бировало клеточную пролиферацию и секрецию клетками ММР-9, а также значительно снижало инвазивную способность миофибробластов. Высокая экспрессия одного из типов рецепто- ров к TNFα на одних и тех же клетках приводит к развитию различных эффектов при действии цитокина. Было показано, что после трансплан- тации клеток костного мозга LDLR-мышам с но- каутом гена рецептора липопротеинов низкой плотности от двух групп мышей (дефицитных по гену TNFR1 и мышей дикого типа, несущих TNFR1 на клеточной мембране) не наблюдалось различий между полученными группами в уров- не холестерина и триглицеридов в крови, а также в общей площади атеросклеротической бляшки (количество Т-лимфоцитов, нейтрофилов, апоп- тотических ядер и коллагена сходно в обеих груп- пах). Однако у химерных мышей с нокаутом гена TNFR1 отмечалось уменьшение макрофагальной площади бляшки, снизилась способность макро- фагов поглощать модифицированные ЛПНП, а также уменьшилась экспрессия данными клет- ками моноцитарного хемотаксического проте- ина 1 (МСР-1), способствующего привлечению новых моноцитов крови в очаг воспаления [69]. у ф р Изменение экспрессии рецепторов к TNFα су- щественно влияет и на приживаемость тканей при их трансплантации. Рецепторы семейства TNF и рецепторы смерти (death receptors) В частности, высокая экспрессия рецеп- тора на мононуклеарных клетках крови (в основ- ном на В-лимфоцитах, имеющих более высокую плотность рецептора) способствует еще боль- шей инфильтрации железы этими клетками при тиреоидите Хашимото (фолликулярная инфиль- трация) и болезни Грейвса (диффузная инфиль- трация) [10]. ка или раком молочной железы ухудшает течение заболевания и способствует прогрессированию болезни, в результате чего снижается общая вы- живаемость и выживаемость без прогрессиро- вания заболевания у пациентов. Увеличение количества DR5 и FasR на мембране раковых клеток повышает резистентность опухоли к хи- миотерапии при рецидиве болезни (но не влияет на лечение первичного заболевания) [17]. Повы- шение экспрессии BCMA (рецептор для BAFF) на клетках хронического лимфолейкоза снижает длительность периода выживаемости без про- грессирования заболевания и увеличивает риск развития рецидива болезни [19]. Сигнализация через Fas-рецептор (CD95, Apo-1) является одним из ключевых механизмов подавления опухолевого роста. Как отмечается в литературных источниках, восприимчивость к CD95-опосредованному апоптозу в значитель- ной степени снижается во многих злокачествен- ных опухолях, что может играть важную роль в опухолевой прогрессии, иммунном уклонении опухолевых клеток, а также устойчивости к хи- миотерапии. Даже клетки, по всем показателям принадлежащие к одному типу опухоли, но по- лученные от разных пациентов, различаются по степени экспрессии FasR. Поэтому от плот- ности рецепторов на клетке и будет зависеть ее ответ при действии FasL. При исследовании плотности экспрессии рецепторов TNFα в норме, у больных ревмато- идным артритом [3, 34], больных атопическим дерматитом [33] и больных активных туберку- лезом легких [2] было показано, что интактные клетки субпопуляций МНК ПК различаются как по относительному проценту клеток, экс- прессирующих рецепторы 1 и 2 типа к TNFα, так и по абсолютному числу рецепторов на них. При этом увеличение или уменьшение плотно- сти экспрессии рецепторов на клетках при пато- логии по сравнению с нормой не всегда проис- ходит с изменением процента клеток, несущих данные рецепторы. В частности, как у здоровых, так и у больных РА вне зависимости от активно- сти воспалительного процесса для субпопуляции Т-лимфоцитов характерен наибольший процент клеток, экспрессирующих рецептор 2 типа, при наименьшей плотности экспрессии рецепторов обоих типов на них. Это может свидетельствовать о том, что для клеток данной субпопуляции пре- дельно допустимая плотность рецепторов низ- кая, но при этом данная популяция клеток явля- ется более чувствительной к действию медиатора и имеющаяся плотность является достаточной для связывания рецепторов с цитокинами и ре- ализации эффектов TNFα на них. Рецепторы семейства TNF и рецепторы смерти (death receptors) По мнению Niederkorn J.Y., пересадка роговицы – старейшая, наиболее рас- пространенная и, как правило, наиболее удачная форма аллотрансплантации твердой ткани. Не- смотря на то, что HLA-соответствие не всегда выполняется, не используются системные им- мунодепрессанты, в 90% случаев первичная ал- лотрансплантация роговицы имеет успех. В ис- следовании проведена трансплантация роговицы от двух групп дефицитных мышей (нокаутирован- ных по гену либо TNFR1, либо TNFR2) в перед- нюю камеру глаза нормальных мышей. Отторже- ние трансплантатов от TNFR1-нокаутированных мышей наблюдалось в 53% случаев. В противо- положность этому, все роговичные трансплан- таты от TNFR2-нокаутировванных мышей были отторгнуты (100%). Таким образом, отсутствие 2 типа рецептора к TNFα повышает риск отторже- ния трансплантата роговицы. Это также свиде- тельствует, что передача сигналов через TNFR2 на донорских клетках способствует развитию раз- личных иммунорегуляторных событий, которые предотвращают индукцию аллоиммунитета [47]. Роль TNFα и его рецепторов в контроле жиз- неспособности клеток в физиологических и па- тологических состояниях до сих пор остается предметом дискуссий. Condorelli и соавт. [13] об- наружили, что недифференцированные клетки нейробластомы человека (линия клеток SK-N- BE) экспрессируют и TNFR1, и TNFR2 в срав- нимых количествах и при действии TNFα по- вышают свою пролиферативную активность. Дифференцировка этих клеток, индуцируемая ретиноевой кислотой (РК), резко повышала экс- прессию 1 типа рецептора при том же уровне экспрессии TNFR2. При дальнейшем культиви- ровании клеток с TNFα происходило ингибиро- вание их роста, что обнаруживалось с помощью оценки повышенного уровня каспазы-3. Блока- да 2 типа рецептора на недифференцированных клетках приводила к снижению клеточной про- лиферации, а на дифференцированных – к уси- лению ингибирующего эффекта. Блокада TNFR1 сопровождалась повышением пролиферации недифференцированных клеток и снижением ингибирующего эффекта на дифференцирован- ных клетках. По мнению авторов, митогенный эффект, индуцированный TNFα в недифферен- цированных клетках, может быть результатом сбалансированной активации обоих типов ре- Экспрессия рецепторов семейства TNF может меняться в ходе неопластических процессов. Так, повышение экспрессии DR4 и TNFR2 на опухо- левых клетках у пациентов с карциномой яични- 382 2019, Vol. 21, 3 2019, Т. 21, № 3 Рецепторы к иммунорегуляторам Receptors for immune regulators болевания. Sellam J. и соавт. показывают, что снижение уровня экспрессии рецептора BAFF на В-лимфоцитах способствует повышению ак- тивности заболевания у больных СКВ (по оценке SLEDAI) и развитию внежелезистых поражений у пациентов с первичным синдромом Шегре- на [57]. Изменение экспрессии BAFF-R может играть роль в развитии заболеваний щитовидной железы. и плотность экспрессии рецепторов на клетке ре- гулируются различными механизмами. ного эффекта в клетке зависят, в том числе, и от компонентов, формирующих цитокин-рецеп- торный комплекс. Таким образом, изменение уровня экспрессии рецепторов семейства TNF приводит к различ- ным ответам клеток при воздействии соответ- ствующего цитокина: изменение интенсивности пролиферации клеток, секреции ими различных медиаторов, индукция апоптоза и др. Если это происходит в условиях патологии, то эффект цитокина на клетки с измененным уровнем экс- прессии рецепторов будет определять исход за- болевания, а в некоторых случаях повышенная или пониженная экспрессия рецептора может стать дополнительным диагностическим крите- рием активности заболевания. При обобщении литературных данных о вли- янии плотности экспрессии рецепторов к раз- личным интерлейкинам было выявлено, что из- менение уровня экспрессии рецепторов имеет большое значение в оценке функционального ответа клетки на медиатор и в развитии патоло- гических состояний (табл. 1). В исследованиях подтверждены данные о влиянии плотности ре- цепторов на процессы пролиферации и апоптоза, а также обменные процессы, что является тригге- ром в развитии аутоиммунных, онкологических и дистрофических заболеваний. Для всех рассмотренных классов регулятор- ных молекул характерным является изменение плотности экспрессии рецепторов как одного из ключевых моментов регулирования функци- ональной активности клеток. Таким образом, изучение уровня экспрессии рецепторов на мем- бране клеток является важным в понимании па- тогенеза, а изменение уровня экспрессии может рассматриваться как терапевтическая мишень в лечении различных заболеваний. Заключение При связывании различных цитокинов с ре- цепторами эффекты, оказываемые на клетку, моделируются несколькими вариантами взаимо- действий. При этом конечный эффект на клетку может определяться изменениями на различ- ных этапах взаимодействия медиатора и клетки (рис. 1). В то же время стандартно определяемый уровень растворимых факторов является лишь одним из многих параметров, регулирующих данные взаимодействия. Поскольку уровень плотности экспрессии рецепторов к факторам роста имеет наибольшее значение в развитии онкологических процессов, этот механизм важно учитывать, так как он вли- яет на трансформацию клетки, а также течение Благодаря этому запуск разных сигнальных путей и, как следствие, достижение определен- Рисунок 1. Факторы, определяющие тип и выраженность эффектов медиатора на клетку-мишень Figure 1. Рецепторы семейства TNF и рецепторы смерти (death receptors) Также во всех исследованных группах выявляется ряд четких закономерностей по субпопуляциям с наиболь- шим процентом клеток, экспрессирующих ре- цепторы 2 типа, и по субпопуляциям с мини- мальной и максимальной плотностью экспрессии рецепторов обоих типов на клетках. Выявленные особенности распределения рецепторов по суб- популяциям позволяют говорить о наличии по- пуляционно-зависимой экспрессии, характери- зующей рецепторный аппарат TNFα, и о том, что изменение процента клеток в субпопуляции, экспрессирующих соответствующий рецептор, Zoi-Toli O. и соавт. в своих исследованиях по- казали, что высокая экспрессия FasR на опухоле- вых клетках кожной В-клеточной [72] и кожной Т-клеточной [73] лимфом была связана с умень- шением прогрессии опухоли, со снижением ри- ска развития рецидивов заболевания и смерти, а пациенты в этом случае имели более благо- приятный исход болезни. Исследование различ- ной плотности экспрессии FasR на опухолевых клетках человеческого рака почки показало, что степень дифференцировки опухоли была обрат- но пропорциональна экспрессии FasR: чем выше плотность рецептора на клетках, тем менее, диф- ференцирована опухоль [52]. Противоположный результат был получен на образцах немелко- клеточного рака легких (НМРЛ): в бронхо-аль- веолярной карциноме, являющейся наиболее дифференцированной из всех типов НМРЛ, на- блюдалась наиболее высокая степень экспрессии FasR [31]. Авторы отмечают также, что пациен- ты с низкоэкспрессирующими типами НМРЛ имели более короткий период выживаемости и более высокую стадию заболевания (по TMN- классификации). Говоря о роли рецепторов семейства TNF, следует затронуть еще одну группу патологи- ческих процессов, таких как аутоиммунные за- 383 Sennikov S.V. et al. Сенников С.В. и др. Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология и плотность экспрессии рецепторов на клетке ре- гулируются различными механизмами. и плотность экспрессии рецепторов на клетке ре- гулируются различными механизмами. Заключение Factors determining the type and intensity of effects of a mediator on the target cell Плотность экспрессии каждого из типов рецепторов на клетке Expression density of each receptor type on the cell Уровень растворимых рецепторов и цитокина The levels of soluble receptors and cytokine Уровень не-Ig растворимых ингибиторов и антагонистов (рецепторные антагонисты и др.) The level of non-Ig soluble inhibitors and antagonists (receptor antagonists etc.) Уровень аутоантител к медиатору Autoantibody level Соотношение количества рецепторов различных типов к одному медиатору на клетке The level of receptors of different types per mediator on the cell Относительное и абсолютное содержание клеток, экспрессирующих те или иные рецепторы The relative and absolute cell counts for cells expressing a certain receptor type Эффект медиатора на клетку-мишень Effect of mediator on the target cell Внутриклеточные белки целевой клетки Intracellular proteins of the target cell Компоненты взаимодействия, формирующие медиатор- рецепторный комплекс The components of interaction forming the mediator-receptor complex Распределение различных типов рецепторов между субпопуляциями (при взаимодействии с мембраносвязанными формами медиаторов на таргетной клетке) Distribution of various types of receptors between subpopulations (upon interaction with membrane- bound forms of mediators on the target cells) Плотность экспрессии каждого из типов рецепторов на клетке Expression density of each receptor type on the cell Уровень растворимых рецепторов и цитокина The levels of soluble receptors and cytokine Плотность экспрессии каждого из типов рецепторов на клетке Expression density of each receptor type on the cell Уровень растворимых рецепторов и цитокина The levels of soluble receptors and cytokine Распределение различных типов рецепторов между субпопуляциями (при взаимодействии с мембраносвязанными формами медиаторов на таргетной клетке) Distribution of various types of receptors between subpopulations (upon interaction with membrane- bound forms of mediators on the target cells) Уровень не-Ig растворимых ингибиторов и антагонистов (рецепторные антагонисты и др.) The level of non-Ig soluble inhibitors and antagonists (receptor antagonists etc.) Эффект медиатора на клетку-мишень Effect of mediator on the target cell Внутриклеточные белки целевой клетки Intracellular proteins of the target cell Уровень аутоантител к медиатору Autoantibody level Компоненты взаимодействия, формирующие медиатор- рецепторный комплекс The components of interaction forming the mediator-receptor complex Соотношение количества рецепторов различных типов к одному медиатору на клетке The level of receptors of different types per mediator on the cell Относительное и абсолютное содержание клеток, экспрессирующих те или иные рецепторы The relative and absolute cell counts for cells expressing a certain receptor type Рисунок 1. Факторы, определяющие тип и выраженность эффектов медиатора на клетку-мишень Figure 1. Заключение Factors determining the type and intensity of effects of a mediator on the target cell 384 Рецепторы к иммунорегуляторам Receptors for immune regulators 2019, Vol. 21, 3 2019, Т. 21, № 3 ТАБЛИЦА 1. ВЗАИМОСВЯЗЬ МЕЖДУ ИЗМЕНЕНИЯМ В ЭКСПРЕССИИ РЕЦЕПТОРОВ К ОСНОВНЫМ КЛАССАМ РЕГУЛЯТОРНЫХ МЕДИАТОРОВ И ЭФФЕКТАМИ НА ТАРГЕТНЫЕ КЛЕТКИ TABLE 1. Заключение RELATIONSHIP BETWEEN CHANGES IN THE EXPRESSION OF RECEPTORS TO THE MAIN CLASSES OF REGULATORY MEDIATORS AND EFFECTS ON TARGET CELLS Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Интерлейкины Interleukins IL-20R Кератиноциты Keratinocytes Повышение плотности – нарушение пролифе- рации и дифференцировки [67] Increased expression – altered proliferation and differentiation [67] IL-20R Остеокласты Osteoclasts Снижение экспрессии – более высокая мине- ральная плотность костной ткани, больший объем костной ткани и толщина губчатой кости [27] Decreased expression – higher bone mineral density, greater volume of bone tissue and thicker cancellous bone [27] IL-31ra Кератиноциты Keratinocytes Повышение плотности – атопический дерматит, псориаз [20] Increased expression – atopic dermatitis, psoriasis [20] IL-1R1, IL-1R2 МНК перифериче- ской крови Peripheral blood mononuclear cells Изменение плотности и процента – атопический дерматит, ревматоидный артрит; изменения плотности и процента – индексы активности заболеваний [4, 5] Changed expression – atopic dermatitis, rheumatoid arthritis; association between the changes and disease activity score [4, 5] IL-4R Моноциты и Т-хелперы Monocytes and T helper cells from newborn umbilical cord blood and peripheral blood one year later Отсутствие снижения плотности в течение первого года жизни – развитие атопического дерматита [21] Absence of decrease of expression during first year after birth – associated with the development of atopic dermatitis [21] IL-1R2 Эпителиальные клетки толстой кишки Colonic epithelial cells Повышение экспрессии – злокачественная трансформация клеток и более тяжелые фор- мы колоректального рака [38] Increased expression – malignant cell transformation and the more severe forms of colorectal cancer [38] IL-4R Наивные B-клетки Naïve B cells Высокая экспрессия α-субъединицы – улучше- ние выживаемости и снижение апоптоза [65] Increased expression – better survival and reduced apoptosis [65] IL-6R Клетки рака поджелудочной железы Pancreatic cancer cells Высокий уровень экспрессии – повышение инвазивной способности клеток [11] Increased expression – enhanced invasiveness of the cells [11] Рецепторы к иммунорегуляторам Рецепторы к иммунорегуляторам Receptors for immune regulators Повышение экспрессии – злокачественная трансформация клеток и более тяжелые фор- мы колоректального рака [38] Increased expression – malignant cell transformation and the more severe forms of colorectal cancer [38] Высокая экспрессия α-субъединицы – улучше- ние выживаемости и снижение апоптоза [65] Increased expression – better survival and reduced apoptosis [65] 385 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. et al. Сенников С.В. и др. Sennikov S.V. et al. Заключение Medical Immunology (Russia)/Meditsinskaya Immunologiya Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Интерлейкины Interleukins IL-6R CD14+ клетки перитонеальной жидкости боль- ных эндометри- озом CD14+ cells in peritoneal fluid of patients with endometriosis Повышенная экспрессия – степень и тяжесть патологии [30] Increased expression – severity of the pathology [30] IL-21R Теноциты Tenocytes Высокая экспрессия – запуск провоспалитель- ных механизмов [9] Increased expression – induction of proinflammatory mechanisms [9] IL-21R Регуляторные Т-клетки в модели человеческого рассеянного склероза – экс- периментальный аутоиммунный энцефаломиелит (ЭАЭ) Regulatory T cells in the model of human multiple sclerosis: experimental autoimmune encephalomyelitis (EAE) Снижение экспрессии – более раннее начало и более тяжелые признаки заболевания, однако более быстрое выздоровление [32] Decreased expression – earlier onset and more severe disease signs but contributes to earlier recovery [32] IL-10R CD4+ и CD8+ Т-клетки CD4+ and CD8+ T cells Повышенная экспрессия – более благоприят- ное течение СКВ [15] Increased expression is associated with a more favorable course of systemic lupus erythematosus (SLE) [15] IL-1R2 Линия HUC-1 HUC-1 cell line Повышенная экспрессия – снижение экспрес- сии корецептора IL-1RAcP и в результате по- давление ответа клетки при действии IL-1 через IL-1R1 [12] Increased expression – reduces expression of IL-1RAcP coreceptor, thus suppressing cell response to IL-1 through IL-1R1 [12] IL-33R Эндотелиальные клетки сосудов Vascular endothelial cells Повышенная экспрессия – прогрессирование атеросклеротических поражений сосудов [39] Increased expression – progression of atherosclerotic vascular lesions [39] Интерфероны Interferons IFNAR-2 Клетки фибросар- комы; рак подже- лудочной железы Fibrosarcoma cells; pancreatic cancer cells Пониженная экспрессия – повышение чувстви- тельности к действию IFNβ, снижение антипро- лиферативного эффекта, при сохранении анти- вирусного эффекта вплоть до снижения уровня экспрессии рецепторов на 50% от исходного. Повышенная экспрессия – более выраженный ответ на действие IFNα [44, 56] Decreased expression – increased sensitivity to IFNβ, while cells with high expression respond more readily to IFNα. A decreased expression – reduced antiproliferative effect but the antiviral effect was maintained at a high level until the level of receptor expression decreased to 50% of the initial level [44, 56] 386 2019, Vol. 21, 3 2019, Т. 21, № 3 Рецепторы к иммунорегуляторам Receptors for immune regulators 387 Receptors for immune regulators 2019, Vol. Заключение 21, 3 Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Интерфероны Interferons IFNGR CD4+ Т-лимфоциты CD4+Т cells Пол-ассоциированные изменения уровня экспрессии – тяжесть протекания бронхиаль- ной астмы [28] Sex-related alterations in the expression level – severity of the course of asthma [28] IFNAR-1 Колоректальный рак Colorectal cancer cells Снижение плотности экспрессии по сравнению с нормальными клетками эпителия слизистой [29] Reduced expression density compared to that in normal mucosal epithelial cells [29] Факторы роста Growth factors EGFR Эпителиальные клетки Epithelial cells Высокая плотность – снижение безрецидивной выживаемости и повышение риска смерти при раке мочевого пузыря, молочной железы, при плоскоклеточном раке пищевода, плоскокле- точном раке головы и шеи, глиоме, при раке желудка, оральном плоскоклеточном в отличие от пациентов с низким уровнем экспрессии ре- цептора на клетках. Уровень экспрессии EGFR прямо пропорцио- нален инвазивной способности клеток при пло- скоклеточном раке гортани и первичном раке эндометрия [37, 42, 43, 46, 48, 53, 54, 55, 71] Increased expression – decreased recurrence-free survival and increased mortality risk in patients with bladder cancer, breast cancer, esophageal squamous cell cancer, head and neck squamous cell carcinoma, glioma, stomach cancer, oral squamous cell carcinoma as opposed to those with a low level of receptor expression on cells. Заключение Cell invasiveness was directly proportional to the EGFR expression level (laryngeal squamous cell carcinoma, primary endometrial carcinoma) [ 37, 42, 43, 46, 48, 53, 54, 55, 71] HER-3 Клетки рака желудка Gastric cancer cells Гиперэкспрессия – ухудшение состояния больных и снижение выживаемости [7, 23] Increased expression – deterioration of patient’s condition and reduced survival [7, 23] HER-2 Клетки аденокар- циномы простаты Prostate adenocarcinoma cells Высокая экспрессия – уровень сывороточного простатического специфического антигена [70] Increased expression – serum level of prostate- specific antigen [70] НЕР2/НЕР3 Клетки рака молочной железы Breast cancer cells Наличие гетеродимеров – усиление пролифе- рации опухолевых клеток [36] The presence of heterodimers – enhanced tumor cell proliferation [36] VEFGR-2, VEGFR-3 VEGF-2, VEGF-3 Клетки эндометрия Endometrial cells Повышение экспрессии – опухолевая транс- формация клеток, повышение ангиогенеза и опухолевой прогрессии; высокая экспрессия VEGFR-3 – повышение миграционной активно- сти клеток и инвазии в миометрий [66] Increased expression – tumor transformation of cells, increased angiogenesis and tumor progression; high level of VEGFR-3 expression is associated with enhanced migration activity of the cells and their myometrial invasion [66] , , , ] Increased expression – decreased recurrence-free survival and increased mortality risk in patients with bladder cancer, breast cancer, esophageal squamous cell cancer, head and neck squamous cell carcinoma, glioma, stomach cancer, oral squamous cell carcinoma as opposed to those with a low level of receptor expression on cells. Cell invasiveness was directly proportional to the EGFR expression level (laryngeal squamous cell carcinoma, primary endometrial carcinoma) [ 37, 42, 43, 46, 48, 53, 54, 55, 71] Повышение экспрессии – опухолевая транс- формация клеток, повышение ангиогенеза и опухолевой прогрессии; высокая экспрессия VEGFR-3 – повышение миграционной активно- сти клеток и инвазии в миометрий [66] Increased expression – tumor transformation of cells, increased angiogenesis and tumor progression; high level of VEGFR-3 expression is associated with enhanced migration activity of the cells and their myometrial invasion [66] 387 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. et al. Сенников С.В. и др. 388 Sennikov S.V. et al. Заключение Medical Immunology (Russia)/Meditsinskaya Immunologiya Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Факторы роста Growth factors VEGF-1 Клетки первично- го рака яичников Primary ovarian cancer cells Повышение экспрессии – повышение риска ре- цидива заболевания после удаления опухоли [68] Increased expression – increased risk of disease recurrence after tumor resection [68] VEGF-1, VEGF-2 VEGF-1, VEGF-2 Макрофаги, эн- дотелиальные клетки Macrophages, endothelial cells Повышение экспрессии – рекрутирование макрофагов в атеросклеротические бляшки; стимулирование англиогенеза сосудов с по- вышенной проницаемостью, что способствует появлению кровоизлияний и экстравазации воспалительных клеток [63] Increased expression – recruitment of macrophages into atherosclerotic plaques; stimulation of angiogenesis of vessels with enhanced permeability, which leads to hemorrhage and inflammatory cell extravasation [63] VEGF-1, VEGF-2 VEGF-1, VEGF-2 Нейроны, эндоте- лиальные клетки и астроциты Neurons, endothelial cells, and astrocytes Изменение экспрессии – болезнь Альцгеймера [22] Altered expression – Alzheimer’s disease [22] c-Met (рецептор фактора роста гепатоцитов) c-Met (hepatocyte growth factor receptor) Эпителиальные клетки при карци- номе носоглотки Epithelial cells in nasopharyngeal carcinoma Повышение уровня экспрессии – повышение двигательной активности раковых клеток, способствует их метастазированию в шейные лимфатические узлы [35] Increased expression – increases motility of cancer cells, thus leading to their metastases into cervical lymph nodes [35] TGF-β-R1 Клетки хрящевой ткани Сartilage tissue cells Возрастное снижение экспрессии – риск разви- тия остеоартрита [64] Age-associated decrease of expression is a risk factor for development of osteoarthritis [64] Рецептор тром- боцитарного фактора роста (PDGFR) Platelet- derived growth factor receptor (PDGFR) Мезенхимальные стволовые клеток Mesenchymal stem cells Низкая плотность экспрессии – снижение ми- грационной способности, дифференцировки в эластин- и актин-продуцирующие фибробла- сты, нарушение ангиогенеза [50] Decreased expression – reduction of migration ability, differentiation of the cells into elastin- and actin-producing fibroblasts; disruption of angiogenesis [50] α-субъединица рецептора GM-CSF α-subunit of receptor to GM-CSF Моноциты Monocytes Пороговый уровень экспрессии – направление дифференцировки (в ДК или макрофаги) [14] Threshold level of expression determines the direction of differentiation (into dendritic cells or macrophages) [14] Повышение экспрессии – рекрутирование макрофагов в атеросклеротические бляшки; стимулирование англиогенеза сосудов с по- вышенной проницаемостью, что способствует появлению кровоизлияний и экстравазации воспалительных клеток [63] Increased expression – recruitment of macrophages into atherosclerotic plaques; stimulation of angiogenesis of vessels with enhanced permeability, which leads to hemorrhage and inflammatory cell extravasation [63] 388 2019, Vol. 21, 3 2019, Т. Заключение 21, № 3 Рецепторы к иммунорегуляторам Receptors for immune regulators Receptors for immune regulators 2019, Vol. Заключение et al. Сенников С.В. и др. 390 Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Рецепторы семейства TNF и рецепторы смерти TNF family receptors and death receptors TNFR2 Роговица TNFR2- нокаутированных мышей Cornea in TNFR2- knockout mice Отсутствие – повышенный риск отторжения трансплантата роговицы [47] Absence – increases the risk of corneal graft rejection [47] DR4, TNFR2 DR4, TNFR2 Опухолевые клет- ки карциномы яичника или рака молочной железы Ovarian carcinoma cells or breast cancer cells Повышение экспрессии – ухудшение течения заболевания, прогрессирование болезни, по- вышение резистентности опухоли к химиотера- пии при рецидиве болезни [17] Increased expression – aggravates the course and leads to disease progression, thus reducing the overall and progression-free survival in patients; increases tumor resistance to chemotherapy in disease relapse but does not affect treatment of the primary disease [17] BCMA Клетки хрониче- ского лимфолей- коза Сhronic lymphocytic leukemia cells Повышение экспрессии – снижение длительно- сти периода выживаемости без прогрессирова- ния заболевания и увеличение риска развития рецидива болезни [19] Increased expression – shortens the progression- free survival period and increases the risk of disease recurrence [19] FasR Опухолевые клетки кожной В-клеточной и кожной Т-клеточной лимфом Cutaneous B-cell and cutaneous T-cell lymphoma cells Высокая экспрессия – уменьшение прогрессии опухоли, снижение риска развития рецидивов заболевания и смерти [72, 73] Increased expression – reduced tumor progression, reduced risks of disease recurrence and death. Patients had a more favorable outcome in this case [72,73] FasR Рак почки Kidney cancer Степень экспрессии FasR – обратно пропор- циональная степени дифференцировки опухо- ли [52] The degree of tumor differentiation was inversely proportional to FasR expression: the higher receptor density on the cells – the more poorly differentiated tumor [52] FasR Немелкоклеточ- ный рак легких (НМРЛ) Non-small cell lung cancer (NSCLC) cells Высокая экспрессия – наиболее дифферен- цированные формы НМРЛ. Низкая экспрес- сия – более короткий период выживаемости и более высокая стадия заболевания (по TMN- классификации) [31] The highest FasR expression level was observed in bronchoalveolar carcinoma, which is the most well-differentiated type of NSCLC. Заключение 21, 3 Иммуно- регуляторные молекулы Immunoregulatory molecules Рецептор Receptor Клетки Cells Ассоциация изменения экспрессии с эффектом/патологией Association between change in expression and effect or pathology Хемокиновые рецепторы Chemokine receptors CXCR7, CCR7, CCR8, CXCR4 Опухолевые клет- ки различного генеза Different-genesis tumor cells Увеличение экспрессии – образование псевдоподий, увеличение подвижности, как следствие – инвазия и метастазирование в лимфоидные органы клеток меланомы кожи, цервикальная интраэпителиальная неоплазия (предраковое состояние плоскоклеточного рака шейки матки), рак молочной железы, плоскоклеточный рак пищевода, желудка [16, 40, 45, 61, 62] Increased expression – leads to formation of pseudopods, higher motility and, therefore, invasion and metastases of skin melanoma cells into the lymphoid organs, cervical intraepithelial neoplasia (precancerous lesion of cervical squamous cell carcinoma), breast cancer, esophageal squamous cell carcinoma, and squamous cell carcinoma of the stomach [16, 40, 45, 61, 62] CX3CR1 Клетки колорек- тального рака Colorectal cancer cells Снижение экспрессии – уменьшение общей вы- живаемости и увеличение частоты прогресси- рования [18] Decreased expression – reduced overall survival and increased progression rate [18] Корецепторы CCR5 и CXCR4 Co-receptors CCR5 and CXCR4 CD4-позитивные клетки организма CD4-positive cells of the body Высокая экспрессия – способствует проник- новению и репликации макрофаготропного и Т-клеточнотропного ВИЧ-1 [24] Increased expression – entry and replication of macrophage-tropic and T cell-tropic HIV-1 [24] CX3CR1 Клетки атеро- склеротической бляшки (сонные артерии) Cells derived from atherosclerotic plaques (carotid arteries) Изменение экспрессии предиктор нестабиль- ности атеросклеротической бляшки [41] Altered expression – predict atherosclerotic plaque instability [41] Рецепторы семейства TNF и рецепторы смерти TNF family receptors and death receptors TNFR1, TNFR2 TNFR1, TNFR2 МНК ПК Peripheral blood mononuclear cells Изменение экспрессии – интегральные показа- тели активности заболевания при РА (DAS-28) и АД (SCORAD), тяжесть туберкулезного про- цесса [2, 3, 25, 33, 34, 58] Changed expression – integral disease activity score in rheumatoid arthritis (DAS-28) and atopic dermatitis (SCORAD) and with severity of tuberculosis [2, 3, 25, 33, 34, 58] TNFR1, TNFR2 TNFR1, TNFR2 Недифференци- рованные клетки нейробластомы человека (линия клеток SK-N-BE) Undifferentiated human neuroblastoma cells (SK-N-BE cell line) Изменение соотношения на клетках – влияние на пролиферацию и дифференцировку [13] The changes in TNFR1/TNFR2 proportion in cells – affected proliferation and differentiation [13] Изменение экспрессии предиктор нестабиль- ности атеросклеротической бляшки [41] Altered expression – predict atherosclerotic plaque instability [41] ц [ , , , , , ] Changed expression – integral disease activity score in rheumatoid arthritis (DAS-28) and atopic dermatitis (SCORAD) and with severity of tuberculosis [2, 3, 25, 33, 34, 58] Изменение соотношения на клетках – влияние на пролиферацию и дифференцировку [13] The changes in TNFR1/TNFR2 proportion in cells – affected proliferation and differentiation [13] Изменение соотношения на клетках – влияние на пролиферацию и дифференцировку [13] The changes in TNFR1/TNFR2 proportion in cells – affected proliferation and differentiation [13] 389 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. Заключение Д (E) Д (E) Е (F) Е (F) Д (E) Таким образом, одним из важных механиз- мов регуляции эффективности действия цитоки- нов и вариабельности их биологических эффек- тов является плотность экспрессии рецепторов на поверхности клеток и количество рецепторов, равноправно участвующих во взаимодействии лиганда с клеткой. Поэтому при исследовании изменений в системе мембраносвязанных форм рецепторов и самого цитокина необходимо оце- нивать не только процентное содержание клеток, несущих рецепторы, но и количество самих экс- прессируемых рецепторов. Рисунок 2. Варианты регуляции эффектов на клетку в зависимости от порогового уровня рецепторов и медиатора Рисунок 2. Варианты регуляции эффектов на клетку в зависимости от порогового уровня рецепторов и медиатора Рисунок 2. Варианты регуляции эффектов на клетку в зависимости от порогового уровня рецепторов и медиатора Примечание. A, Б – отсутствие эффекта в связи с выраженным дефицитом одного из компонентов взаимодействия; В, Г – слабый сигнал / первый тип сигналинга; Д, Е – более сильный сигнал / второй тип сигналинга из-за достижения порогового уровня связывания для определенного рецептора. Figure 2. The regulation of cellular effects depends on the relationship between the soluble cytokine and the density of receptors expressed on cells Figure 2. The regulation of cellular effects depends on the relationship between the soluble cytokine and the density of receptors expressed on cells Изучение плотности экспрессии рецепторов к медиаторам имеет важное практическое зна- чение для понимания механизмов ответа клетки при изменении уровня экспрессии рецепторов в норме и при различных патологиях. Все это мо- жет служить терапевтической мишенью для лече- ния и профилактики различных заболеваний. Note. A, B, no effect on the cell or week signaling because one of the interaction components is missing; С, D, typical signaling; E, F, forced signaling/another type of signaling achieves the threshold binding level for a certain receptor. Note. A, B, no effect on the cell or week signaling because one of the interaction components is missing; С, D, typical signaling; E, F, forced signaling/another type of signaling achieves the threshold binding level for a certain receptor. Заключение Patients with low- expressing types of NSCLC had a shorter survival period and a higher grade (according to the TMN system) [31] Рецептор BAFF BAFF receptor В-лимфоциты B cells Снижение экспрессии – повышение активности заболевания у больных СКВ (по оценке SLEDAI) и развитие внежелезистых поражений у паци- ентов с первичным синдромом Шегрена [57] Reduced expression increases of SLEDAI score for disease activity in SLE patients and leads to development of extraglandular lesions in patients with primary Sjögren syndrome [57] Повышение экспрессии – снижение длительно- сти периода выживаемости без прогрессирова- ния заболевания и увеличение риска развития рецидива болезни [19] Increased expression – shortens the progression- free survival period and increases the risk of disease recurrence [19] Высокая экспрессия – уменьшение прогрессии опухоли, снижение риска развития рецидивов заболевания и смерти [72, 73] Increased expression – reduced tumor progression, reduced risks of disease recurrence and death. Patients had a more favorable outcome in this case [72,73] Снижение экспрессии – повышение активности заболевания у больных СКВ (по оценке SLEDAI) и развитие внежелезистых поражений у паци- ентов с первичным синдромом Шегрена [57] Reduced expression increases of SLEDAI score for disease activity in SLE patients and leads to development of extraglandular lesions in patients with primary Sjögren syndrome [57] 390 2019, Vol. 21, 3 2019, Т. 21, № 3 Рецепторы к иммунорегуляторам Рецепторы к иммунорегуляторам Receptors for immune regulators Receptors for immune regulators и исход заболеваний, диагностику и их лечение. А (A) Б (B) В (C) Г (D) Д (E) Е (F) А (A) Изменение плотности экспрессии хемокиновых рецепторов на клетках может определять мор- фологические и функциональные изменения в клетках и, как следствие, развитие патологиче- ского процесса. Б (B) Изменение уровня экспрессии рецепторов семейства TNF приводит к различным ответам клеток при воздействии соответствующего цито- кина: изменение интенсивности пролиферации клеток, секреции ими различных медиаторов, индукция апоптоза и др. В некоторых случаях повышенная или пониженная экспрессия ре- цептора может стать дополнительным диагно- стическим критерием активности заболевания или определять его исход. В (C) Г (D) Г (D) В (C) Рецепторы на клетках и сам медиатор являют- ся равноценными участниками взаимодействия. Поскольку для ряда растворимых форм регу- ляторов показано, что существуют пороговые уровни для ответа клетки на действие цитокина и/или для изменения эффекта на клетку, логич- но предположить, что аналогичные пороговые уровни существуют и для экспрессии рецепторов на клетках (рис. 2). Тогда отношение плотности экспрессии и уровень растворимого цитокина являются фактором, определяющим тип и выра- женность эффектов на клетку. 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Authors: Sennikov S.V., PhD, MD (Medicine), Professor, Head, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology; Novosibirsk State University, Novosibirsk, Russian Federation Список литературы / References Expression of Fas and Fas- ligand in primary cutaneous T-cell lymphoma (CTCL): association between lack of Fas expression and aggressive types of CTCL. Br. J. Dermatol., 2000, Vol. 143, no. 2, pp. 313-319. Alshevskaya A.A., PhD (Medicine), Research Associate, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation y pp 69. Xanthoulea S., Gijbels M.J., van der Made I., Mujcic H., Thelen M., Vergouwe M.N., Ambagts M.H., Hofker M.H., de Winther M.P. P55 tumour necrosis factor receptor in bone marrow-derived cells promotes atherosclerosis development in low-density lipoprotein receptor knock-out mice. Cardiovasc. Res., 2008, Vol. 80, no. 2, pp. 309-318. t Авторы: Сенников С.В. – д.м.н., профессор, заведующий лабораторией молекулярной иммунологии ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии»; ФГАОУ ВО «Новосибирский национальный исследовательский государственный университет», г. Новосибирск, Россия Alshevskaya A.A., PhD (Medicine), Research Associate, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation Zhukova Yu.V., Postgraduate Student, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation 395 Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Sennikov S.V. et al. Сенников С.В. и др. Sennikov S.V. et al. Сенников С.В. и др. Medical Immunology (Russia)/Meditsinskaya Immunologiya Медицинская Иммунология Belomestnova I.A., Student, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation Беломестнова И.А. – студентка лаборатории молекулярной иммунологии ФГБНУ «Научно- исследовательский институт фундаментальной и клинической иммунологии», г. Новосибирск, Россия Karaulov A.V., PhD, MD (Medicine), Professor, Head, Department of Clinical Immunology and Allergology, First Moscow I. Sechenov State Medical University, Moscow, Russian Federation Karaulov A.V., PhD, MD (Medicine), Professor, Head, Department of Clinical Immunology and Allergology, First Moscow I. Sechenov State Medical University, Moscow, Russian Federation Караулов А.В. – д.м.н., профессор, заведующий кафедрой клинической иммунологии и аллергологии ФГАОУ ВО «Первый Московский государственный медицинский университет имени И.М. Сеченова» Министерства здравоохранения РФ (Сеченовский университет), Москва, Россия Lopatnikova Yu.A., PhD (Medicine), Senior Research Associate, Laboratory of Molecular Immunology, Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russian Federation Лопатникова Ю.А. – к.м.н., старший научный сотрудник лаборатории молекулярной иммунологии ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии», г. Новосибирск, Россия Поступила 05.03.2019 Отправлена на доработку 05.04.2019 Принята к печати 15.05.2019 Received 05.03.2019 Revision received 05.04.2019 Accepted 15.05.2019 Received 05.03.2019 Revision received 05.04.2019 Accepted 15.05.2019 396	19,735
https://openalex.org/W4290180089	OpenAlex	Open Science	CC-By	2,022	Multiple types of navigational information are independently encoded in the population activities of the dentate gyrus neurons	Tomoyuki Murano	English	Spoken	35,181	63,998	. CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Multiple types of navigational information are independently encoded in the population 1 activities of the dentate gyrus neurons 2 3 Tomoyuki Murano1, Ryuichi Nakajima1, Akito Nakao1,2, Nao Hirata1, Satoko 4 Amemori1,3, Akira Murakami4, Yukiyasu Kamitani4, Jun Yamamoto5, Tsuyoshi Miyakawa1* 5 6 7 1Division of Systems Medical Science, Institute for Comprehensive Medical Science, 8 Fujita Health University, Toyoake, Japan 9 2Department of Synthetic Chemistry and Biological Chemistry, Graduate School of 10 Engineering, Kyoto University, Kyoto, Japan 11 3Division of Systems Neuroscience, Primate Research Institute, Kyoto University, 12 Inuyama, Japan 13 4Graduate School of Informatics, Kyoto University, Yoshida-honmachi, Sakyo-ku, 14 Kyoto 606-8501, Japan 15 5Department of Psychiatry, University of Texas Southwestern Medical Center, Dallas, 16 Texas, USA 17 18 19 Correspondence: 20 Tsuyoshi Miyakawa 21 Division of Systems Medical Science, Institute for Comprehensive Medical Science, 22 Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, Aichi 470-1192, 23 Japan 24 Telephone: +81-562-93-9376 25 Fax: +81-562-92-5382 26 E-mail: miyakawa@fujita-hu.ac.jp 27 28 Multiple types of navigational information are independently encoded in the population 1 activities of the dentate gyrus neurons 2 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Abstract 29 30 The dentate gyrus (DG) plays critical roles in cognitive functions such as learning, memory, 31 and spatial coding, and its dysfunction is implicated in various neuropsychiatric disorders. 32 However, it remains largely unknown how information is represented in this region. Here, we 33 recorded neuronal activity in the DG using Ca2+ imaging in freely moving mice and analysed 34 this activity using machine learning. The activity patterns of populations of DG neurons 35 enabled us to successfully decode position, speed, and motion direction in an open field as well 36 as current and future location in a T-maze, and each individual neuron was diversely and 37 independently tuned to these multiple information types. In αCaMKII heterozygous knockout 38 mice, which present deficits in spatial remote and working memory, the decoding accuracy of 39 position in the open field and future location in the T-maze were selectively reduced. These 40 results suggest that multiple types of information are independently distributed in DG neurons. 41 42 Abstract 29 30 The dentate gyrus (DG) plays critical roles in cognitive functions such as learning, memory, 31 and spatial coding, and its dysfunction is implicated in various neuropsychiatric disorders. 32 However, it remains largely unknown how information is represented in this region. Here, we 33 recorded neuronal activity in the DG using Ca2+ imaging in freely moving mice and analysed 34 this activity using machine learning. The activity patterns of populations of DG neurons 35 enabled us to successfully decode position, speed, and motion direction in an open field as well 36 as current and future location in a T-maze, and each individual neuron was diversely and 37 independently tuned to these multiple information types. Multiple types of navigational information are independently encoded in the population 1 activities of the dentate gyrus neurons 2 In αCaMKII heterozygous knockout 38 mice, which present deficits in spatial remote and working memory, the decoding accuracy of 39 position in the open field and future location in the T-maze were selectively reduced. These 40 results suggest that multiple types of information are independently distributed in DG neurons. 41 42 42 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Introduction 43 44 The dentate gyrus (DG) in the hippocampus has been implicated in cognitive 45 functions such as pattern separation1,2, contextual encoding3,4, and place recognition3,4,7–9, and 46 its dysfunction is suggested to be associated with various neuropsychiatric disorders such as 47 epilepsy10, schizophrenia11,12, bipolar disorder11,13, and Alzheimer’s disease10. The neural 48 circuitry of DG is composed of excitatory input from layer II neurons in the entorhinal cortex 49 (EC)14,15; local interactions among DG granule cells (GCs), mossy cells, and interneurons4,7,8; 50 and excitatory output to CA3 pyramidal cells4,7. The EC receives multimodal sensory 51 information from other brain regions and contains neurons with distinct functional properties, 52 including grid16, border17, speed18,19, and head-direction cells20. Likewise, the hippocampal CA 53 regions receive information from the DG and contains neurons that represent place21, 54 locomotion speed19, episodic memory22, time23, and novel spatial experience24,25. Therefore, 55 the DG is thought to be involved in the processing and integration of a variety of information26. 56 57 Introduction 43 In spite of its wide-ranging functions, the DG, is unusual among hippocampal regions 58 in that only a small population of principal DG neurons is thought to be active in a given 59 environment27,28. In addition, several studies have reported that most DG neurons exhibit poor 60 tuning to position (e.g., DG neurons often have multiple and irregular firing fields3,29) and 61 movement speed (e.g., most DG neurons are not specifically tuned to running or resting 62 states30). This paradox has led to a significant interest in how information is represented in the 63 population activity patterns of DG neurons9. 64 Here, to unveil important functional characteristics of DG neurons, we performed 66 Ca2+ imaging using a micro-endoscope in freely moving mice, which enable us to record 67 activity from a large population of neurons31,32. We recorded neural activity of the dorsal DG, 68 that is thought to be relatively more involved in the exploratory behaviour and contextual 69 memory encoding than in ventral DG5. Then, we analysed how individual DG neurons are 70 involved in encoding position, speed, and motion direction in an open field as well as current 71 and future location (left or right) in a T-maze. Even if the majority of individual neurons are 72 poorly tuned to these types of information, neurons may encode information by the population 73 activity patterns9. Therefore, by using machine learning methods, we also asked if these types 74 of information are encoded in the population activity patterns of DG neurons. Concurrently, to 75 assess how these neural representations might be altered in a disease model, we carried out the 76 same imaging and analysis in heterozygous alpha-calcium/calmodulin-dependent kinase II 77 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint knockout (αCaMKII+/–) mice. Results 88 89 1. Activity patterns of dorsal DG neurons are tuned to the position and speed of the mouse. 90 By using a head-mounted miniaturized microscope, we recorded neural activity in 91 the upper blade of the dorsal DG (dDG) of freely moving mice while they were travelling in 92 an open field (Fig. 1a). We observed that over 90% of all GCaMP expressing neurons in the 93 field of view were active in both of wild-type and αCaMKII+/– mice (Supplementary Fig. 1), 94 and included 72.6±9.9 dDG neurons from 7 wild-type mice and 57.8±9.6 dDG neurons from 5 95 αCaMKII+/– mice into analysis (for details see “Detection of Ca2+ transients” in Materials and 96 Methods section). 97 We analysed the tuning specificities of single dDG neurons regarding mice position, 98 speed, and motion direction. Representative spatial tuning maps, speed tuning curves, and 99 direction tuning maps of individual dDG neurons are shown in Figure 1c. We computed 100 information content (bits/calcium transient)44 about the position, speed, and motion direction 101 for each neuron, which we designated spatial, speed, and direction information, respectively. 102 In wild-type mice, the average of spatial information of the dDG neurons was significantly 103 larger than that expected by chance (permutation test, P < 10–3) and the effect size was medium 104 (Cohen’s d = 0.53). The distribution of the spatial information was shifted to be slightly larger 105 than that of the shuffled data (Fig. 1d), and the significance of spatial information for each cell 106 varied widely from small to large (Supplementary Fig. 2), indicating that most of the dDG 107 neurons are tuned to information about position to varying degrees. The same was also true of 108 the amount of speed information (P < 10–3; d = 0.54; Fig. 1d). The amount of direction 109 information was also larger than that expected by chance (P = 3.00 × 10–3; Fig. 1d), the effect 110 size was small (d = 0.11; Fig. 1d), suggesting that the activity patterns of dDG neurons are 111 relatively weakly tuned to motion direction. Similarly, in αCaMKII+/– mice, the spatial and 112 speed information of dDG neurons were significantly greater than would be expected by 113 chance (spatial, P < 10–3, d = 0.38; speed, P < 10–3, d = 0.58; Fig. 1d), but direction information 114 was not (P = 0.03, d = 0.01; Fig. 1d). Results 88 Thus, in both wild-type and αCaMKII+/– mice, individual 115 dDG neurons are tuned to position and speed to varying degrees, but are relatively weakly 116 tuned to motion direction. Indeed, it appears that these information types are widely distributed 117 in the populations of dDG neurons. 118 Introduction 43 Mutations of αCaMKII have been reported to cause intellectual 78 disability accompanied by epilepsy, abnormal emotional and affective behaviour, and/or 79 autistic features33–36, and the gene has been also implicated in autism37 and bipolar disorder38,39. 80 The αCaMKII+/– mice exhibit an array of behavioural abnormalities, including locomotor 81 hyperactivity, impaired working and remote memory, abnormal social/aggressive behaviour, 82 and exaggerated infradian rhythms39–43. By studying how neural representation of information 83 is altered in the DG of these mice, we further aimed to gain insight into how multiple types of 84 information are represented in the population activities of dDG neurons in normal and 85 abnormal conditions. 86 87 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 2. Information about position, speed, and motion direction is encoded in the population 121 activity patterns of dDG neurons. 122 1. Activity patterns of dorsal DG neurons are tuned to the position and speed of the mouse. 90 On 139 the other hand, the decoding accuracy of position was better than chance to a marginally 140 significant degree (P = 0.0549), but was significantly lower than that of wild-type mice 141 (unpaired t-test, Obs.–Shuff. is compared; P = 1.12 × 10–3; Fig. 2d). This lower position 142 decoding accuracy in αCaMKII+/– mice compared to wild-type mice might not be due to the 143 difference in the number of the neurons used for decoding between them (position decoding 144 accuracy × the number of neurons, R = –0.0279; Supplementary Fig. 7). Thus, in αCaMKII+/– 145 mice, position information is selectively impaired in the dDG, whereas speed and motion 146 direction information is not. These results imply that with respect to information about speed 147 and motion direction, information about position is encoded in the dDG by a different 148 population of neurons, a different coding principle, or both. 149 150 151 3 Information about position speed and motion direction is independently distributed in 152 Next, to examine the information coding at the population level, we tested whether 123 position, speed, and motion direction of the mice could be decoded from the population activity 124 of dDG neurons using machine learning methods. Eight models were considered for decoding 125 analysis in this study, and since there was no significant difference in the decoding accuracy 126 among these models, we used the LSTM (Long Short-Term Memory), which showed a slightly 127 higher accuracy than others (Supplementary Fig. 3). Representative decoding results are shown 128 together with the actual positions, speeds, and motion directions in Figure 2a–c (Decoding 129 errors for all mice are shown in Supplementary Fig. 4–6). In wild-type mice, the decoding 130 accuracies for position, speed, and motion direction were significantly higher than those 131 expected by chance, which is the decoding accuracy when using the shuffled data (paired t-test, 132 position, P = 2.18 × 10–4; speed, P = 2.42 × 10–3; motion direction, P = 1.36 × 10–3; Fig. 2d–f, 133 left panels). These results indicate that information about position, speed, and motion direction 134 are encoded in the population activity of dDG neurons in wild-type mice. 1. Activity patterns of dorsal DG neurons are tuned to the position and speed of the mouse. 90 135 In αCaMKII+/– mice, the decoding accuracies for speed and motion direction were 136 significantly higher than those expected by chance (paired t-test, speed, P = 1.66 × 10–2; motion 137 direction, P = 0.0210), and not significantly different from those of wild-type mice (unpaired 138 t-test, Obs.–Shuff. is compared; speed, P = 0.619; motion direction, P = 0.262; Fig. 2e, 2f). On 139 the other hand, the decoding accuracy of position was better than chance to a marginally 140 significant degree (P = 0.0549), but was significantly lower than that of wild-type mice 141 (unpaired t-test, Obs.–Shuff. is compared; P = 1.12 × 10–3; Fig. 2d). This lower position 142 decoding accuracy in αCaMKII+/– mice compared to wild-type mice might not be due to the 143 difference in the number of the neurons used for decoding between them (position decoding 144 accuracy × the number of neurons, R = –0.0279; Supplementary Fig. 7). Thus, in αCaMKII+/– 145 mice, position information is selectively impaired in the dDG, whereas speed and motion 146 direction information is not. These results imply that with respect to information about speed 147 and motion direction, information about position is encoded in the dDG by a different 148 population of neurons, a different coding principle, or both. 149 1. Activity patterns of dorsal DG neurons are tuned to the position and speed of the mouse. 90 2. Information about position, speed, and motion direction is encoded in the population 121 activity patterns of dDG neurons. 122 2. Information about position, speed, and motion direction is encoded in the population 121 activity patterns of dDG neurons. 122 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Next, to examine the information coding at the population level, we tested whether 123 position, speed, and motion direction of the mice could be decoded from the population activity 124 of dDG neurons using machine learning methods. Eight models were considered for decoding 125 analysis in this study, and since there was no significant difference in the decoding accuracy 126 among these models, we used the LSTM (Long Short-Term Memory), which showed a slightly 127 higher accuracy than others (Supplementary Fig. 3). Representative decoding results are shown 128 together with the actual positions, speeds, and motion directions in Figure 2a–c (Decoding 129 errors for all mice are shown in Supplementary Fig. 4–6). In wild-type mice, the decoding 130 accuracies for position, speed, and motion direction were significantly higher than those 131 expected by chance, which is the decoding accuracy when using the shuffled data (paired t-test, 132 position, P = 2.18 × 10–4; speed, P = 2.42 × 10–3; motion direction, P = 1.36 × 10–3; Fig. 2d–f, 133 left panels). These results indicate that information about position, speed, and motion direction 134 are encoded in the population activity of dDG neurons in wild-type mice. 135 In αCaMKII+/– mice, the decoding accuracies for speed and motion direction were 136 significantly higher than those expected by chance (paired t-test, speed, P = 1.66 × 10–2; motion 137 direction, P = 0.0210), and not significantly different from those of wild-type mice (unpaired 138 t-test, Obs.–Shuff. is compared; speed, P = 0.619; motion direction, P = 0.262; Fig. 2e, 2f). 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 3c, middle 187 panel), showing that the amount of speed information in a neuron is also independent from the 188 f i l di i i f i F di i d di h i ifi 189 more types of information than others, or the amount of one type of information carried by a 158 neuron may be independent from that of other types of information. 159 neuron may be independent from that of other types of information. 159 At first, to clarify the relationship among the amount of information encoded by 160 individual neurons for these three information types, we examined correlations between them. 161 Scatter plots for each pair of variables (among position, speed, and direction information) show 162 fairly uniform distributions and relatively weak correlations (spatial × speed, R = 0.167; speed 163 × direction, R = –1.67 × 10–4; direction × spatial, R = 0.257; Fig. 3a). These weak correlations 164 suggest that there is limited correspondence between how each form of information is encoded 165 in individual neurons. 166 Next, we examined whether populations of dDG neurons used for coding position, 167 speed, and motion direction are exclusive from, identical to, or independent of each other, by 168 assessing how the decoding performance changes when neurons are deleted from the datasets 169 one by one. In our analysis, when neurons were deleted from the dataset in order of decreasing 170 spatial information, position decoding accuracy decreased more rapidly than when they were 171 deleted randomly (Two-way ANOVA with Tukey-Kramer post hoc tests, Spatial info order × 172 Random order, P = 2.29 × 10–10; Fig. 3c, left panel); the reverse was true of deletion in order 173 of increasing spatial information (Reverse spatial info order × Random order, P = 5.20 × 10–4; 174 Fig. 3c, left panel). These results indicate that neuron populations consisting of individual 175 neurons with larger amounts of spatial information tend to be more important in the population 176 coding of position information than those with smaller amount of spatial information. 177 Furthermore, the removal of neurons in decreasing speed or direction information order 178 mimicked the impacts of randomly ordered deletion on position decoding performance (Fig. 179 3c, left panel). These results support the notion that the information about position is 180 independently encoded from that of speed or motion direction. 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 Here, we investigated how each neuron encodes position, speed, and motion 154 direction. One possibility is that a neuron may carry a particular type of information exclusively. 155 Another possibility is that a neuron may carry multiple types of information. In this case, some 156 particular groups of neurons may carry a relatively larger amount of information about two or 157 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint more types of information than others, or the amount of one type of information carried by a 158 neuron may be independent from that of other types of information. 159 At first, to clarify the relationship among the amount of information encoded by 160 individual neurons for these three information types, we examined correlations between them. 161 Scatter plots for each pair of variables (among position, speed, and direction information) show 162 fairly uniform distributions and relatively weak correlations (spatial × speed, R = 0.167; speed 163 × direction, R = –1.67 × 10–4; direction × spatial, R = 0.257; Fig. 3a). These weak correlations 164 suggest that there is limited correspondence between how each form of information is encoded 165 in individual neurons. 166 Next, we examined whether populations of dDG neurons used for coding position, 167 speed, and motion direction are exclusive from, identical to, or independent of each other, by 168 assessing how the decoding performance changes when neurons are deleted from the datasets 169 one by one. 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 In our analysis, when neurons were deleted from the dataset in order of decreasing 170 spatial information, position decoding accuracy decreased more rapidly than when they were 171 deleted randomly (Two-way ANOVA with Tukey-Kramer post hoc tests, Spatial info order × 172 Random order, P = 2.29 × 10–10; Fig. 3c, left panel); the reverse was true of deletion in order 173 of increasing spatial information (Reverse spatial info order × Random order, P = 5.20 × 10–4; 174 Fig. 3c, left panel). These results indicate that neuron populations consisting of individual 175 neurons with larger amounts of spatial information tend to be more important in the population 176 coding of position information than those with smaller amount of spatial information. 177 Furthermore, the removal of neurons in decreasing speed or direction information order 178 mimicked the impacts of randomly ordered deletion on position decoding performance (Fig. 179 3c, left panel). These results support the notion that the information about position is 180 independently encoded from that of speed or motion direction. Similarly, the removal of 181 neurons in decreasing speed information order lowered speed decoding accuracy to a greater 182 extent than randomly ordered deletion (Speed info order × Random order, P = 1.40 × 10–8; Fig. 183 3c, middle panel); on the other hand, deletion in the reverse order had smaller impacts on speed 184 decoding accuracy (Reverse speed info order × Random order, P = 4.23 × 10–3; Fig. 3c, middle 185 panel). Removal of neurons in increasing spatial or direction information order did not decrease 186 speed decoding performance more rapidly than randomly ordered deletion (Fig. 3c, middle 187 panel), showing that the amount of speed information in a neuron is also independent from the 188 amount of spatial or direction information For direction decoding there was no significant 189 more types of information than others, or the amount of one type of information carried by a 158 neuron may be independent from that of other types of information. 159 At first, to clarify the relationship among the amount of information encoded by 160 individual neurons for these three information types, we examined correlations between them. 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 161 Scatter plots for each pair of variables (among position, speed, and direction information) show 162 fairly uniform distributions and relatively weak correlations (spatial × speed, R = 0.167; speed 163 × direction, R = –1.67 × 10–4; direction × spatial, R = 0.257; Fig. 3a). These weak correlations 164 suggest that there is limited correspondence between how each form of information is encoded 165 in individual neurons. 166 Next, we examined whether populations of dDG neurons used for coding position, 167 speed, and motion direction are exclusive from, identical to, or independent of each other, by 168 assessing how the decoding performance changes when neurons are deleted from the datasets 169 one by one. In our analysis, when neurons were deleted from the dataset in order of decreasing 170 spatial information, position decoding accuracy decreased more rapidly than when they were 171 deleted randomly (Two-way ANOVA with Tukey-Kramer post hoc tests, Spatial info order × 172 Random order, P = 2.29 × 10–10; Fig. 3c, left panel); the reverse was true of deletion in order 173 of increasing spatial information (Reverse spatial info order × Random order, P = 5.20 × 10–4; 174 Fig. 3c, left panel). These results indicate that neuron populations consisting of individual 175 neurons with larger amounts of spatial information tend to be more important in the population 176 coding of position information than those with smaller amount of spatial information. 177 Furthermore, the removal of neurons in decreasing speed or direction information order 178 mimicked the impacts of randomly ordered deletion on position decoding performance (Fig. 179 3c, left panel). These results support the notion that the information about position is 180 independently encoded from that of speed or motion direction. Similarly, the removal of 181 neurons in decreasing speed information order lowered speed decoding accuracy to a greater 182 extent than randomly ordered deletion (Speed info order × Random order, P = 1.40 × 10–8; Fig. 183 3c, middle panel); on the other hand, deletion in the reverse order had smaller impacts on speed 184 decoding accuracy (Reverse speed info order × Random order, P = 4.23 × 10–3; Fig. 3c, middle 185 panel). Removal of neurons in increasing spatial or direction information order did not decrease 186 speed decoding performance more rapidly than randomly ordered deletion (Fig. 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 Similarly, the removal of 181 neurons in decreasing speed information order lowered speed decoding accuracy to a greater 182 extent than randomly ordered deletion (Speed info order × Random order, P = 1.40 × 10–8; Fig. 183 3c, middle panel); on the other hand, deletion in the reverse order had smaller impacts on speed 184 decoding accuracy (Reverse speed info order × Random order, P = 4.23 × 10–3; Fig. 3c, middle 185 panel). Removal of neurons in increasing spatial or direction information order did not decrease 186 speed decoding performance more rapidly than randomly ordered deletion (Fig. 3c, middle 187 panel), showing that the amount of speed information in a neuron is also independent from the 188 amount of spatial or direction information. For direction decoding, there was no significant 189 difference in the decoding accuracy for the removal of neurons in decreasing 190 spatial/speed/direction information order, increasing direction information order, or random 191 order (Fig. 3c, right panel). These results showed that the amount of spatial, speed, or direction 192 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint information in an individual neuron is not associated with its contribution to the decoding 193 accuracy of motion direction. Thus, neuron populations involved in coding these three types of 194 information are not mutually exclusive or identical to each other, but rather are independent of 195 each other. 196 information in an individual neuron is not associated with its contribution to the decoding 193 accuracy of motion direction. Thus, neuron populations involved in coding these three types of 194 information are not mutually exclusive or identical to each other, but rather are independent of 195 each other. 196 information in an individual neuron is not associated with its contribution to the decoding 193 accuracy of motion direction. 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 Then, at first, we examined the 200 relationship between position decoding accuracy and distinctness of activity patterns of dDG 201 neurons across the subareas in the open field. In our data, the population vector overlap (PVO) 202 (see Materials and Methods section; Supplementary Fig. 8a), which measures how similar 203 firing patterns are across different subareas in the open field, was significantly correlated with 204 the error of position decoding (Supplementary Fig. 8b). Thus, the increased position decoding 205 error in αCaMKII+/– mice relative to that in wild-type mice (P = 1.12 × 10–3, Fig. 2d) might be 206 attributable to the larger mean PVO (P = 0.0208, Supplementary Fig. 8b), corresponding to 207 less distinct firing patterns of neural ensembles across different subareas. Next, we investigated 208 how encoding of speed information is associated with overall firing frequency of dDG neurons. 209 We found positive and linear correlations between the speed of the mice and the average Ca2+ 210 transient rate in the dDG (Supplementary Fig. 8c). We also found a significant correlation 211 between speed decoding accuracy and the change in the average Ca2+ transient rate with speed 212 (Supplementary Fig. 8d), neither of which were significantly different between wild-type and 213 αCaMKII+/– mice (Fig. 2e, Supplementary Fig. 8d). Thus, information about position and speed 214 might be encoded with different coding principles in the dDG, and only the coding principle 215 for position may be affected in the dDG of αCaMKII+/– mice. 216 217 218 4. The activity patterns of dDG neurons are tuned to current and future locations to 219 varying degrees. 220 Next, we recorded the activity patterns of dDG neurons while the mice performed a 221 forced-alternation task in a modified T-maze (Fig. 4a; for details, see “Behavioural experiments 222 in Materials and Methods section). We used the T-maze test because it has been used previously 223 to assess working memory in rodents46 and because dysfunction in the DG is associated with 224 reduced performance in this test46. Wild-type mice made the correct choice (that is, during the 225 free-choice period, they chose the arm opposite the arm that was open in the forced-choice 226 period) 71.4% of the time, averaged of 50 sessions. In contrast, the αCaMKII+/– mice showed 227 3. Information about position, speed, and motion direction is independently distributed in 152 the population activity patterns of dDG neurons. 153 Thus, neuron populations involved in coding these three types of 194 information are not mutually exclusive or identical to each other, but rather are independent of 195 each other. 196 Furthermore, as described above, there is another possibility that different types of 197 information are encoded in the dDG by a different coding principle (e.g., in CA1 and EC, 198 position information is represented in the activity patterns of neurons3,45, and speed information 199 is encoded by the frequency of the neural activity18,19). Then, at first, we examined the 200 relationship between position decoding accuracy and distinctness of activity patterns of dDG 201 neurons across the subareas in the open field. In our data, the population vector overlap (PVO) 202 (see Materials and Methods section; Supplementary Fig. 8a), which measures how similar 203 firing patterns are across different subareas in the open field, was significantly correlated with 204 the error of position decoding (Supplementary Fig. 8b). Thus, the increased position decoding 205 error in αCaMKII+/– mice relative to that in wild-type mice (P = 1.12 × 10–3, Fig. 2d) might be 206 attributable to the larger mean PVO (P = 0.0208, Supplementary Fig. 8b), corresponding to 207 less distinct firing patterns of neural ensembles across different subareas. Next, we investigated 208 how encoding of speed information is associated with overall firing frequency of dDG neurons. 209 We found positive and linear correlations between the speed of the mice and the average Ca2+ 210 transient rate in the dDG (Supplementary Fig. 8c). We also found a significant correlation 211 between speed decoding accuracy and the change in the average Ca2+ transient rate with speed 212 (Supplementary Fig. 8d), neither of which were significantly different between wild-type and 213 αCaMKII+/– mice (Fig. 2e, Supplementary Fig. 8d). Thus, information about position and speed 214 might be encoded with different coding principles in the dDG, and only the coding principle 215 for position may be affected in the dDG of αCaMKII+/– mice. 216 217 each other. 196 Furthermore, as described above, there is another possibility that different types of 197 information are encoded in the dDG by a different coding principle (e.g., in CA1 and EC, 198 position information is represented in the activity patterns of neurons3,45, and speed information 199 is encoded by the frequency of the neural activity18,19). 4. The activity patterns of dDG neurons are tuned to current and future locations to . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 52.0% correct choice on the average, which is close to chance (50%) and significantly lower 228 than the performance of the wild-type mice (P = 2.38 × 10–3; Fig. 4b), indicating that the 229 αCaMKII+/– mice showed deficits in this working memory task. 230 Next, to elucidate the functional properties of dDG neurons in T-maze test, we 231 calculated the “LR index”, which takes a higher absolute value if a neuron shows differential 232 activity patterns when the mouse is on either the left or right side of the apparatus. Here, 233 “current location” designates the (left or right) location of the mouse at time of recording. These 234 neural activity patterns were taken from 5 sec after the mouse performed the initial forced turn, 235 which was designated the “forced arm period” (Fig. 4c). “Future location” refers to the (left or 236 right) location of the mouse after the “decision period”, during which the neural activity 237 patterns for calculating the LR index and decoding the future LR location were taken. The 238 “decision period” was defined as the period from 6 to 1 sec prior to the free-choice turn (Fig. 239 4e). Note that in this period, the future location did not significantly correlate with physical 240 location, speed, and motion direction of the mouse (Supplementary Fig. 9g–i). In the wild-type 241 mice, the absolute values of the LR indices of neurons during the forced arm period were 242 significantly higher than those expected by chance (permutation test, P < 10–3) and the effect 243 size was medium (Cohen’s d = 0.43). The distribution of LR indices of neurons was shifted to 244 be slightly larger in absolute values than that of the shuffled data (Fig. 4. The activity patterns of dDG neurons are tuned to current and future locations to 4d, upper panel), 245 showing that the activity patterns of dDG neurons are tuned to left or right of the current 246 location to varying degrees. The same was also true of the LR indices of neurons during the 247 decision period in wild-type mice (P < 10–3, d = 0.52; Fig. 4f, upper panel) and of those in the 248 forced arm and decision period in αCaMKII+/– mice (forced arm period, P < 10–3, d = 0.51; 249 decision period, P < 10–3, d = 0.54; Fig. 4d and 4f, lower panel). These results showed that the 250 activity patterns of dDG neurons of wild-type and αCaMKII+/– mice are tuned to both current 251 and future LR locations to varying degrees. 252 52.0% correct choice on the average, which is close to chance (50%) and significantly lower 228 than the performance of the wild-type mice (P = 2.38 × 10–3; Fig. 4b), indicating that the 229 αCaMKII+/– mice showed deficits in this working memory task. 230 5. Information about current and future LR location is encoded in the population activity 255 patterns of dDG neurons. 256 The scatter plot of the LR indices of the dDG neurons obtained 283 in the forced arm and decision periods of the wild-type mice showed a small but statistically 284 significant correlation (R = 0.2281, P = 7.91 × 10–5; Fig. 5a). Meanwhile, in αCaMKII+/– mice, 285 the LR indices of the neurons obtained in the forced arm and decision periods showed a uniform 286 distribution, and these indices were not significantly correlated (R = –0.1210, P = 0.1687; Fig. 287 5b). Thus, the LR-tunings of dDG neurons about current and future LR locations are weakly 288 correlated in wild-type mice and largely independent of each other in αCaMKII+/– mice. These 289 correlations suggest that there is a significant, but weak, relationship between LR-tunings of 290 individual dDG neurons during the forced arm- and decision-period in wild-type mice. 291 To examine how information about the current and future LR locations is distributed 292 in the population activity patterns of dDG neurons, we assessed how the LR decoding accuracy 293 in the forced arm and decision periods changed when neurons were removed from the datasets 294 one by one. In the data collected in wild-type mice during the arm period, upon deletion of 295 neurons in order of decreasing LR index for the forced arm period, the LR decoding accuracy 296 of the forced arm period decreased more rapidly than it did upon randomly ordered deletion 297 in the decision period more accurately than that expected by chance (P = 0.0117; Fig. 4h, left 263 panels). These results suggest that information about the current and future LR location may 264 be encoded in dDG neuron activity patterns obtained in the current and decision period, 265 respectively. Meanwhile, in αCaMKII+/– mice, the decoding accuracy of the current LR 266 location in the forced arm period was more accurate than that expected by chance (P = 0.0220; 267 Fig. 4g, right panel) and not significantly different from that of wild-type mice (unpaired t-test, 268 Obs–Shuf. is compared, P = 0.125; Fig. 4g), indicating that information about current LR 269 location is encoded in dDG neuron activity patterns of αCaMKII+/– mice. We also aimed to 270 estimate the future LR location of αCaMKII+/– mice from the population activity of the decision 271 period, but its accuracy was not higher than that expected by chance (P = 0.873; Fig. 5. Information about current and future LR location is encoded in the population activity 255 patterns of dDG neurons. 256 4h, right 272 panel) and was significantly lower than that of wild-type mice (P = 1.19 × 10–4; Fig. 4h), 273 suggesting that future LR choice is not accurately represented in dDG neurons in αCaMKII+/– 274 mice. Thus, the neural representation of future location is selectively impaired in the dDG of 275 αCaMKII+/– mice, whereas that of the current location is not, implying that current and future 276 locations in the T-maze may be independently encoded among the populations of dDG neurons. 277 278 5. Information about current and future LR location is encoded in the population activity 255 patterns of dDG neurons. 256 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint in the decision period more accurately than that expected by chance (P = 0.0117; Fig. 4h, left 263 panels). These results suggest that information about the current and future LR location may 264 be encoded in dDG neuron activity patterns obtained in the current and decision period, 265 respectively. Meanwhile, in αCaMKII+/– mice, the decoding accuracy of the current LR 266 location in the forced arm period was more accurate than that expected by chance (P = 0.0220; 267 Fig. 4g, right panel) and not significantly different from that of wild-type mice (unpaired t-test, 268 Obs–Shuf. is compared, P = 0.125; Fig. 4g), indicating that information about current LR 269 location is encoded in dDG neuron activity patterns of αCaMKII+/– mice. We also aimed to 270 estimate the future LR location of αCaMKII+/– mice from the population activity of the decision 271 period, but its accuracy was not higher than that expected by chance (P = 0.873; Fig. 4h, right 272 panel) and was significantly lower than that of wild-type mice (P = 1.19 × 10–4; Fig. 4h), 273 suggesting that future LR choice is not accurately represented in dDG neurons in αCaMKII+/– 274 mice. Thus, the neural representation of future location is selectively impaired in the dDG of 275 αCaMKII+/– mice, whereas that of the current location is not, implying that current and future 276 locations in the T-maze may be independently encoded among the populations of dDG neurons. 277 278 279 6. Information about current and future LR locations is widely distributed in the 280 population activity patterns of dDG neurons 281 Next, we investigated the relationship between LR indices in forced arm and decision 282 periods of each dDG neurons. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 5e), showing 323 that the LR decoding accuracy during the decision period was higher, PVO tend to be smaller, 324 meaning that neuronal activity patterns in these periods are in more contrasting patterns (i.e., 325 if a neuron is relatively active in one period, it becomes relatively inactive in another period). 326 These results indicate that information about future LR location is more accurately represented 327 in the dDG when there is a stronger association and contrasting patterns between the population 328 activity of dDG neurons in the forced arm and decision periods. 329 330 (Two-way ANOVA with Tukey-Kramer post hoc tests, P = 1.54 × 10–6; Fig. 5c, left panel); 298 deletion of neurons in order of increasing LR index had smaller impacts on the LR decoding 299 accuracy than randomly ordered deletion (P = 1.12 × 10–5; Fig. 5c, left panel). However, the 300 removal of neurons in order of increasing LR index for the decision period did not decrease 301 decoding accuracy more rapidly than randomly ordered deletion (Fig. 5c, left panel). 302 Meanwhile, for LR decoding in the decision period, the deletion of neurons in order of 303 increasing LR index for the decision period order significantly decreased the decoding 304 accuracies (P = 1.92 × 10–2; Fig. 5c, right panel), and reverse order deletion showed smaller 305 decreases than that by random order deletion (P = 1.27 × 10–2; Fig. 5c, right panel). On the 306 other hand, the removal of neurons in order of decreasing LR index for the forced arm period 307 did not have much impact on LR decoding performance, similar to the impact of randomly 308 ordered deletion (Fig. 5c, right panel). These results suggest that neuron population involved 309 in coding current location is independent from that of future location, and vice versa. Thus, 310 information about the current and future location is expected to be independently distributed 311 in the population activity patterns of dDG neurons. 312 As described above, we noted a small but significant correlation between the LR 313 indices of the forced arm and decision periods in wild-type mice (R = 0.2281, P = 7.91 × 10–5; 314 Fig. 5a). We sought to examine whether associations between neural activity patterns during 315 the forced arm and decision periods correlate with an accurate neural representation of future 316 LR location during the decision period. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 Next, we investigated the relationship between LR indices in forced arm and decision 282 periods of each dDG neurons. The scatter plot of the LR indices of the dDG neurons obtained 283 in the forced arm and decision periods of the wild-type mice showed a small but statistically 284 significant correlation (R = 0.2281, P = 7.91 × 10–5; Fig. 5a). Meanwhile, in αCaMKII+/– mice, 285 the LR indices of the neurons obtained in the forced arm and decision periods showed a uniform 286 distribution, and these indices were not significantly correlated (R = –0.1210, P = 0.1687; Fig. 287 5b). Thus, the LR-tunings of dDG neurons about current and future LR locations are weakly 288 correlated in wild-type mice and largely independent of each other in αCaMKII+/– mice. These 289 correlations suggest that there is a significant, but weak, relationship between LR-tunings of 290 individual dDG neurons during the forced arm- and decision-period in wild-type mice. 291 To examine how information about the current and future LR locations is distributed 292 in the population activity patterns of dDG neurons, we assessed how the LR decoding accuracy 293 in the forced arm and decision periods changed when neurons were removed from the datasets 294 one by one. In the data collected in wild-type mice during the arm period, upon deletion of 295 neurons in order of decreasing LR index for the forced arm period, the LR decoding accuracy 296 of the forced arm period decreased more rapidly than it did upon randomly ordered deletion 297 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint (Two-way ANOVA with Tukey-Kramer post hoc tests, P = 1.54 × 10–6; Fig. 5c, left panel); 298 deletion of neurons in order of increasing LR index had smaller impacts on the LR decoding 299 accuracy than randomly ordered deletion (P = 1.12 × 10–5; Fig. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 5c, left panel). However, the 300 removal of neurons in order of increasing LR index for the decision period did not decrease 301 decoding accuracy more rapidly than randomly ordered deletion (Fig. 5c, left panel). 302 Meanwhile, for LR decoding in the decision period, the deletion of neurons in order of 303 increasing LR index for the decision period order significantly decreased the decoding 304 accuracies (P = 1.92 × 10–2; Fig. 5c, right panel), and reverse order deletion showed smaller 305 decreases than that by random order deletion (P = 1.27 × 10–2; Fig. 5c, right panel). On the 306 other hand, the removal of neurons in order of decreasing LR index for the forced arm period 307 did not have much impact on LR decoding performance, similar to the impact of randomly 308 ordered deletion (Fig. 5c, right panel). These results suggest that neuron population involved 309 in coding current location is independent from that of future location, and vice versa. Thus, 310 information about the current and future location is expected to be independently distributed 311 in the population activity patterns of dDG neurons. 312 As described above, we noted a small but significant correlation between the LR 313 indices of the forced arm and decision periods in wild-type mice (R = 0.2281, P = 7.91 × 10–5; 314 Fig. 5a). We sought to examine whether associations between neural activity patterns during 315 the forced arm and decision periods correlate with an accurate neural representation of future 316 LR location during the decision period. First, we calculated the PVO (see Materials and 317 Methods section) of the averaged neuronal activity frequency between the forced arm and 318 decision periods for each mouse. We next compared them with LR decoding accuracy during 319 the decision period. We here defined decoding accuracy as the difference between observed 320 and shuffled data (Obs–Shuff) to standardize, because decoding accuracy expected by chance 321 in decision period can be larger than 50% due to the bias of LR choice of mouse (Fig. 4h). Our 322 result showed that there was a significant negative correlation (R = –0.8738; Fig. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 Furthermore, in αCaMKII+/– mice, which present deficits in spatial remote and 337 working memory, the neural representation of information about position in the open field and 338 future LR location in the T-maze was selectively impaired, supporting the notion that different 339 types of information are independently distributed among dDG neurons. 340 Discussion 331 332 In this study, we demonstrated that multiple types of information (position, speed, 333 and motion direction in an open field as well as current and future left or right location in a T- 334 maze) were represented in the population activity patterns of dDG neurons. These multiple 335 types of information were encoded by overlapping but independent populations of dDG 336 neurons. Furthermore, in αCaMKII+/– mice, which present deficits in spatial remote and 337 working memory, the neural representation of information about position in the open field and 338 future LR location in the T-maze was selectively impaired, supporting the notion that different 339 types of information are independently distributed among dDG neurons. 340 341 The accurate decoding of position (in the open field) and current LR location (in the 342 T-maze) in wild-type mice indicates that the DG is involved in place coding, which is 343 consistent with previous studies3,4,7,8. Additionally, the successful decoding of speed and 344 motion direction in the open field and future location in the T-maze in this study indicates that 345 the dDG is also involved in the neural representation of these types of information, which only 346 a few studies have reported30,9. It is of interest that the decoding accuracies in our study were 347 accurate despite the DG neurons is generally thought to have a lower range of firing frequencies 348 compared to that of most other brain regions28. Recently, recordings using tetrode7, silicon 349 probe4, and calcium imaging8 showed that the activity patterns of GCs were at much lower 350 frequency than those of DG mossy cells and CA3 pyramidal cells4,7,8,28. In our data, the average 351 recorded Ca2+ transient frequency was quite low, approximately 0.02 Hz, which was 352 comparable to that of GCs (1–2/min) reported by the previous imaging study8. Our results thus 353 suggest that the dDG neurons may be involved in information coding with their low-frequency 354 activity patterns. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 First, we calculated the PVO (see Materials and 317 Methods section) of the averaged neuronal activity frequency between the forced arm and 318 decision periods for each mouse. We next compared them with LR decoding accuracy during 319 the decision period. We here defined decoding accuracy as the difference between observed 320 and shuffled data (Obs–Shuff) to standardize, because decoding accuracy expected by chance 321 in decision period can be larger than 50% due to the bias of LR choice of mouse (Fig. 4h). Our 322 result showed that there was a significant negative correlation (R = –0.8738; Fig. 5e), showing 323 that the LR decoding accuracy during the decision period was higher, PVO tend to be smaller, 324 meaning that neuronal activity patterns in these periods are in more contrasting patterns (i.e., 325 if a neuron is relatively active in one period, it becomes relatively inactive in another period). 326 These results indicate that information about future LR location is more accurately represented 327 in the dDG when there is a stronger association and contrasting patterns between the population 328 activity of dDG neurons in the forced arm and decision periods. 329 330 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Discussion 331 332 In this study, we demonstrated that multiple types of information (position, speed, 333 and motion direction in an open field as well as current and future left or right location in a T- 334 maze) were represented in the population activity patterns of dDG neurons. These multiple 335 types of information were encoded by overlapping but independent populations of dDG 336 neurons. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 355 The accurate decoding of position (in the open field) and current LR location (in the 342 T-maze) in wild-type mice indicates that the DG is involved in place coding, which is 343 consistent with previous studies3,4,7,8. Additionally, the successful decoding of speed and 344 motion direction in the open field and future location in the T-maze in this study indicates that 345 the dDG is also involved in the neural representation of these types of information, which only 346 a few studies have reported30,9. It is of interest that the decoding accuracies in our study were 347 accurate despite the DG neurons is generally thought to have a lower range of firing frequencies 348 compared to that of most other brain regions28. Recently, recordings using tetrode7, silicon 349 probe4, and calcium imaging8 showed that the activity patterns of GCs were at much lower 350 frequency than those of DG mossy cells and CA3 pyramidal cells4,7,8,28. In our data, the average 351 recorded Ca2+ transient frequency was quite low, approximately 0.02 Hz, which was 352 comparable to that of GCs (1–2/min) reported by the previous imaging study8. Our results thus 353 suggest that the dDG neurons may be involved in information coding with their low-frequency 354 activity patterns. 355 We found that the populations of DG neurons cooperate to encode information about 357 position, speed, and motion direction in an open field, indicating that these types of information 358 are diffusely distributed among dDG neurons, which is consistent with a recent report9. In this 359 report, the authors proposed that the interpretability of individual neurons is not necessarily 360 important for their contribution to information coding based on the weak correlation between 361 an index of spatial information and the “importance index”, a measure of the contribution of 362 each neuron to decoding. We evaluated the importance of a neuron in decoding by assessing 363 how the decoding performance changes when a neuron is deleted from the population. In 364 contrast with their conclusion, our results indicate that dDG neurons with a larger amount of 365 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint information are more important for decoding position and speed in an open field than those 366 with smaller amounts of information. In addition, we identified redundancy in the distribution 367 of some types of information in the dDG; our results indicate that only the top 30–40% of all 368 neurons were sufficient for decoding position and speed with maximum accuracy (Fig. 3c; red 369 dotted line in the left panel; blue dotted line in the middle panel). On the other hand, 370 approximately 70–80% of all the recorded neurons can contribute to the accuracy of spatial or 371 speed decoding (Fig. 3c; solid red line in the left panel; solid blue line in the middle panel). 372 The neurons with an intermediate information content, which are not needed for population 373 encoding when the neurons with the largest information content are spared, can contribute to 374 the decoding accuracy when these top-class neurons were deleted, further demonstrating the 375 redundancy of information encoding in the individually recorded neurons. Furthermore, 376 although we recorded only approximately 70–80 neurons on average per wild-type mouse, out 377 of approximately 500,000 neurons in the entire DG47, our decoding, based on the activity 378 patterns of this small number of cells, was fairly accurate. It has been reported that there is a 379 difference in the distribution of functional cell types between the dorsal and ventral DG5; even 380 taking this into account, it is assumed that there are far more neurons in the entire dorsal DG 381 than are needed to accurately encode these types of information. Generally, the number of states 382 that a neural circuit can represent is thought to depend on the number of neurons and the 383 dynamic range of firing frequencies of the neurons28. Given this assumption, we propose that 384 the diffuse, redundant encoding of information in a large number of infrequently firing neurons 385 allows the DG to represent a large number of behavioural states in its population activity 386 patterns. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 This selective impairment is unlikely to be attributed to selective death or 434 general functional impairment of specific groups of neurons because we found that neurons 435 neurons to be tuned to different types of information in patterns that are independent of each 401 other. While a previous report proposed that the groups of neurons encoding position partially 402 overlap with those encoding motion direction9, the dependency/independency of the 403 distribution of these types of information was not evaluated. In our study, we found that 404 multiple types of information are independently distributed among dDG neurons by 405 quantitatively evaluating the importance of the neurons involved in the decoding (Fig. 3c). The 406 selective impairments of information (position in open field and future location in T-maze) in 407 αCaMKII+/– mice also support our claim that multiple types of information are independently 408 distributed. The multiplexity and independence of dDG neurons that we have demonstrated in 409 this study suggests that the involvement of dDG neurons in encoding certain types of 410 information does not prevent them from encoding other types of information. Together with 411 their diffuseness and redundancy, the multiplexity and independence of dDG neurons seem to 412 allow the DG not only to express a large number of states for each type of information but also 413 to express these states for multiple types of information. Other than the information types 414 described in this study, some other types of information are thought to be encoded in the DG, 415 such as vestibular, olfactory, visual, and auditory information26,51, which may also be encoded 416 by taking advantage of the diffuseness, redundancy, and multiplexity of DG neurons. 417 neurons to be tuned to different types of information in patterns that are independent of each 401 other. While a previous report proposed that the groups of neurons encoding position partially 402 overlap with those encoding motion direction9, the dependency/independency of the 403 distribution of these types of information was not evaluated. In our study, we found that 404 multiple types of information are independently distributed among dDG neurons by 405 quantitatively evaluating the importance of the neurons involved in the decoding (Fig. 3c). The 406 selective impairments of information (position in open field and future location in T-maze) in 407 αCaMKII+/– mice also support our claim that multiple types of information are independently 408 distributed. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 387 Remarkably, we also discovered that some of the neurons of the DG demonstrate 389 multiplexity, i.e., each of these neurons is involved in encoding multiple types of information. 390 Neuronal multiplexity has also been reported in other brain regions, including the medial EC, 391 in which some neurons can encode position, speed, and head direction48, and the parietal and 392 frontal cortexes, in which some neurons can encode multiple behavioural or task-relevant 393 variables49,50. Neuronal multiplexities are thought to be essential for representing a large 394 number of independent variables in the population activity patterns of a limited number of 395 neurons49,50. It is known that if the functional tunings for multiple variables in an individual 396 neuron are in patterns similar to those of other neurons, the number of independent variables 397 that can be represented by the population activity patterns will be reduced. This is because the 398 dimensions represented by the collective activity of these neurons would not be larger than that 399 of a single neuron50. In other words, to represent many types of information, it is necessary for 400 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint neurons to be tuned to different types of information in patterns that are independent of each 401 other. While a previous report proposed that the groups of neurons encoding position partially 402 overlap with those encoding motion direction9, the dependency/independency of the 403 distribution of these types of information was not evaluated. In our study, we found that 404 multiple types of information are independently distributed among dDG neurons by 405 quantitatively evaluating the importance of the neurons involved in the decoding (Fig. 3c). 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 The 406 selective impairments of information (position in open field and future location in T-maze) in 407 αCaMKII+/– mice also support our claim that multiple types of information are independently 408 distributed. The multiplexity and independence of dDG neurons that we have demonstrated in 409 this study suggests that the involvement of dDG neurons in encoding certain types of 410 information does not prevent them from encoding other types of information. Together with 411 their diffuseness and redundancy, the multiplexity and independence of dDG neurons seem to 412 allow the DG not only to express a large number of states for each type of information but also 413 to express these states for multiple types of information. Other than the information types 414 described in this study, some other types of information are thought to be encoded in the DG, 415 such as vestibular, olfactory, visual, and auditory information26,51, which may also be encoded 416 by taking advantage of the diffuseness, redundancy, and multiplexity of DG neurons. 417 418 Mutations of αCaMKII have been reported to cause intellectual disability 419 accompanied by epilepsy, abnormal emotional and affective behaviour, and/or autistic 420 features33–36, and the gene has also been implicated in autism37 and bipolar disorder38,39. 421 αCaMKII+/– mice have an endophenotype called “immature dentate gyrus (iDG)”, in which the 422 neurons in the dentate gyrus are in a pseudo-immature status39,42. The phenomenon similar to 423 their iDG can be seen in other mouse models of neuropsychiatric disorders and human 424 patients43,52. A prominent feature of this phenotype is that the induction of expression of 425 immediately early genes, such as c-Fos and Arc, are almost completely absent in the granule 426 cells in the DG42,53–55. Interestingly, despite a dramatic reduction in the expression of c-Fos and 427 Arc, the DG neurons of αCaMKII+/– mice seem to fire in a fairly normal manner with regard 428 to the frequency of calcium transients (Supplementary Fig. 1), indicating that the machinery 429 linking calcium transients and the expression of those genes is disrupted in this phenotype and 430 that the expression of c-Fos or Arc is not always a good index of neuronal firing. Another 431 interesting result obtained from the αCaMKII+/– mice was that the decoding accuracy of 432 position in the open field was selectively impaired in the dDG, while those of speed and 433 direction were not. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 Thus, the selective impairment of position information in the DG of 455 αCaMKII+/– mice may be attributed to an increased excitability and disruption of the organized 456 activity patterns of its neurons. On the other hand, speed information in the dDG of αCaMKII+/– 457 mice was not disturbed, possibly because it depends on changes in the neural firing frequency, 458 which is not affected by increased neuronal excitability. The dDG may utilize different coding 459 principles for encoding different types of information, which supports the notion that individual 460 neurons can independently participate in encoding different types of information. Future 461 studies on the mechanism underlying the selective impairment of information in αCaMKII+/– 462 mice and other model mice that show similar phenotypes would contribute to the understanding 463 of the pathophysiology of neuropsychiatric disorders that share this phenotype. 464 465 carrying position information also encode other types of information in wild-type mice. Instead, 436 we assume that different types of information are encoded by different coding principles and 437 that the coding principle for position information is selectively impaired in the dDG of 438 αCaMKII+/– mice. Previous studies have reported that successful spatial coding relies on having 439 distinct firing patterns of hippocampal neurons for different locations3,45. Our results showed 440 that, similar to the hippocampus, distinct firing patterns across different subareas in the dDG 441 are significantly associated with accurate spatial coding, suggesting that position information 442 might be encoded by the ensemble activities of neurons. On the other hand, it has been reported 443 that neurons in the hippocampus and EC show positive and largely linear firing responses to 444 running speed18,19. In our data, we also found a significant correlation between speed decoding 445 accuracy and the average Ca2+ transient rate. Thus, information about position and speed might 446 be encoded with different coding principles in the dDG, and only the coding principle for 447 position may be affected in the dDG of αCaMKII+/– mice. Furthermore, it was recently reported 448 that the cell death of interneurons between the CA1 region and the DG in epileptic mice causes 449 desynchronized hippocampal activity, which results in poor spatial processing in the 450 hippocampus45. It is also known that hippocampal pyramidal neurons of mice with a point 451 mutation in the αCaMKII show a high firing rate and poor spatial selectivity56. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 The multiplexity and independence of dDG neurons that we have demonstrated in 409 this study suggests that the involvement of dDG neurons in encoding certain types of 410 information does not prevent them from encoding other types of information. Together with 411 their diffuseness and redundancy, the multiplexity and independence of dDG neurons seem to 412 allow the DG not only to express a large number of states for each type of information but also 413 to express these states for multiple types of information. Other than the information types 414 described in this study, some other types of information are thought to be encoded in the DG, 415 such as vestibular, olfactory, visual, and auditory information26,51, which may also be encoded 416 by taking advantage of the diffuseness, redundancy, and multiplexity of DG neurons. 417 Mutations of αCaMKII have been reported to cause intellectual disability 419 accompanied by epilepsy, abnormal emotional and affective behaviour, and/or autistic 420 features33–36, and the gene has also been implicated in autism37 and bipolar disorder38,39. 421 αCaMKII+/– mice have an endophenotype called “immature dentate gyrus (iDG)”, in which the 422 neurons in the dentate gyrus are in a pseudo-immature status39,42. The phenomenon similar to 423 their iDG can be seen in other mouse models of neuropsychiatric disorders and human 424 patients43,52. A prominent feature of this phenotype is that the induction of expression of 425 immediately early genes, such as c-Fos and Arc, are almost completely absent in the granule 426 cells in the DG42,53–55. Interestingly, despite a dramatic reduction in the expression of c-Fos and 427 Arc, the DG neurons of αCaMKII+/– mice seem to fire in a fairly normal manner with regard 428 to the frequency of calcium transients (Supplementary Fig. 1), indicating that the machinery 429 linking calcium transients and the expression of those genes is disrupted in this phenotype and 430 that the expression of c-Fos or Arc is not always a good index of neuronal firing. Another 431 interesting result obtained from the αCaMKII+/– mice was that the decoding accuracy of 432 position in the open field was selectively impaired in the dDG, while those of speed and 433 direction were not. This selective impairment is unlikely to be attributed to selective death or 434 general functional impairment of specific groups of neurons because we found that neurons 435 . CC-BY 4.0 International license perpetuity. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint carrying position information also encode other types of information in wild-type mice. Instead, 436 we assume that different types of information are encoded by different coding principles and 437 that the coding principle for position information is selectively impaired in the dDG of 438 αCaMKII+/– mice. Previous studies have reported that successful spatial coding relies on having 439 distinct firing patterns of hippocampal neurons for different locations3,45. Our results showed 440 that, similar to the hippocampus, distinct firing patterns across different subareas in the dDG 441 are significantly associated with accurate spatial coding, suggesting that position information 442 might be encoded by the ensemble activities of neurons. On the other hand, it has been reported 443 that neurons in the hippocampus and EC show positive and largely linear firing responses to 444 running speed18,19. In our data, we also found a significant correlation between speed decoding 445 accuracy and the average Ca2+ transient rate. Thus, information about position and speed might 446 be encoded with different coding principles in the dDG, and only the coding principle for 447 position may be affected in the dDG of αCaMKII+/– mice. Furthermore, it was recently reported 448 that the cell death of interneurons between the CA1 region and the DG in epileptic mice causes 449 desynchronized hippocampal activity, which results in poor spatial processing in the 450 hippocampus45. It is also known that hippocampal pyramidal neurons of mice with a point 451 mutation in the αCaMKII show a high firing rate and poor spatial selectivity56. Increased 452 excitability and epileptiform activity have also been observed in the DG and CA1 of mice with 453 mutations in αCaMKII+/–, which may lead to disorganized activity patterns in the neurons of 454 these regions57,56,42,58. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 Increased 452 excitability and epileptiform activity have also been observed in the DG and CA1 of mice with 453 mutations in αCaMKII+/–, which may lead to disorganized activity patterns in the neurons of 454 these regions57,56,42,58. Thus, the selective impairment of position information in the DG of 455 αCaMKII+/– mice may be attributed to an increased excitability and disruption of the organized 456 activity patterns of its neurons. On the other hand, speed information in the dDG of αCaMKII+/– 457 mice was not disturbed, possibly because it depends on changes in the neural firing frequency, 458 which is not affected by increased neuronal excitability. The dDG may utilize different coding 459 principles for encoding different types of information, which supports the notion that individual 460 neurons can independently participate in encoding different types of information. Future 461 studies on the mechanism underlying the selective impairment of information in αCaMKII+/– 462 mice and other model mice that show similar phenotypes would contribute to the understanding 463 of the pathophysiology of neuropsychiatric disorders that share this phenotype. 464 465 The traditional view of place coding in the hippocampal circuit is that the place cells 466 encode the current location of the animal. It has also been known that neural activity patterns 467 in CA125, CA359, and medial EC60 are involved in the prospective representation of future states. 468 In our analysis, we were able to decode whether the mice were on the left or right side of the 469 T-maze by the population activity patterns of dDG neurons either after or before a left or right 470 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint turn. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 These results suggest that the population activity patterns of dDG neurons not only encode 471 the current location, but also may be involved in the predictive representation of future states, 472 alongside other hippocampal regions. Furthermore, our result in Fig. 5e showed that this 473 predictive representation of the future states seem to be formed in the population activity 474 pattern of dDG neurons, especially when they are associated with those of the recent past. 475 476 In conclusion, our findings suggest that multiple types of information are diffusely, 477 redundantly, and independently encoded in the population activity patterns of dDG neurons. 478 These features of information coding may be the basis on which the DG is involved in the 479 processing and integration of variety of information. Future studies are needed to determine 480 whether information about other functions of the DG (e.g., episodic memory, object recognition, 481 and odour information processing) is similarly encoded in the population activity patterns of 482 neurons, and how this encoding is altered in disease states (e.g., amnesia, Alzheimer disease, 483 and schizophrenia). 484 485 turn. These results suggest that the population activity patterns of dDG neurons not only encode 471 the current location, but also may be involved in the predictive representation of future states, 472 alongside other hippocampal regions. Furthermore, our result in Fig. 5e showed that this 473 predictive representation of the future states seem to be formed in the population activity 474 pattern of dDG neurons, especially when they are associated with those of the recent past. 475 476 In conclusion, our findings suggest that multiple types of information are diffusely, 477 redundantly, and independently encoded in the population activity patterns of dDG neurons. 478 These features of information coding may be the basis on which the DG is involved in the 479 processing and integration of variety of information. Future studies are needed to determine 480 whether information about other functions of the DG (e.g., episodic memory, object recognition, 481 and odour information processing) is similarly encoded in the population activity patterns of 482 neurons, and how this encoding is altered in disease states (e.g., amnesia, Alzheimer disease, 483 and schizophrenia). 484 485 . CC-BY 4.0 International license perpetuity. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 After the injection, the scalp was sutured and treated with 517 povidone-iodine. DG consists of granule cells (~90%) and contains other types of excitatory 518 neurons, such as mossy cells in hilus and interneurons47. GCaMP6f expression is driven by the 519 Syn-promoter; however, it is not cell type specific. Therefore, its expression is expected to be 520 Materials and Methods 486 487 488 489 Animals 490 All experimental protocols were approved by the Institutional Animal Care and Use Committee 491 of Fujita Health University. Adult male C57BL/6J mice and αCaMKII+/- mice were obtained 492 from Jackson Laboratories (Bar Harbor, Maine) and were backcrossed to C57BL/6J mice 493 (Charles River, Yokohama) for at least 19 generations. They were used for experiments at 50 494 ± 4.4 weeks of age. The mice were housed one per cage in a room with a 12 hr light/dark cycle 495 (lights on at 7:00 a.m., off at 7:00 p.m.) with access to food and water ad libitum. The room 496 temperature was maintained at 23 ± 2 °C. All experiments were conducted during the light 497 period. 498 499 500 501 Surgeries 502 Viral delivery of Ca2+ sensor 503 For delivery of a fluorescent Ca2+ sensor into dorsal DG neurons, adeno-associated virus (AAV) 504 carrying the GCaMP6f vector was injected into the DG of the dorsal hippocampus in adult 505 mice by a conventional method (right hemisphere; 2.0 mm posterior to bregma (AP), 1.0 mm 506 lateral to midline (ML), and 2.0 mm ventral to bregma (DV))61. In brief, adult mice >8 weeks 507 old were first anaesthetized with 1.5–3% isoflurane at an oxygen flow rate of 1 L/min. The 508 head fur was shaved, and the incision site was sterilized with 70% ethanol prior to the beginning 509 of the surgical procedure. The mice were then mounted on a stereotaxic device (51730D, 510 Stoelting, IL), and a heat pad (BWT-100A, BRC, Nagoya) was placed underneath the mouse 511 to maintain body temperature at 37 °C. After the scalp was incised and pulled aside, a 2 mm- 512 diameter craniotomy was created with a surgical drill on the skull above the injection site. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Materials and Methods 486 487 488 489 Animals 490 All experimental protocols were approved by the Institutional Animal Care and Use Committee 491 of Fujita Health University. Adult male C57BL/6J mice and αCaMKII+/- mice were obtained 492 from Jackson Laboratories (Bar Harbor, Maine) and were backcrossed to C57BL/6J mice 493 (Charles River, Yokohama) for at least 19 generations. They were used for experiments at 50 494 ± 4.4 weeks of age. The mice were housed one per cage in a room with a 12 hr light/dark cycle 495 (lights on at 7:00 a.m., off at 7:00 p.m.) with access to food and water ad libitum. The room 496 temperature was maintained at 23 ± 2 °C. All experiments were conducted during the light 497 period. 498 499 500 501 Surgeries 502 Viral delivery of Ca2+ sensor 503 For delivery of a fluorescent Ca2+ sensor into dorsal DG neurons, adeno-associated virus (AAV) 504 carrying the GCaMP6f vector was injected into the DG of the dorsal hippocampus in adult 505 mice by a conventional method (right hemisphere; 2.0 mm posterior to bregma (AP), 1.0 mm 506 lateral to midline (ML), and 2.0 mm ventral to bregma (DV))61. In brief, adult mice >8 weeks 507 old were first anaesthetized with 1.5–3% isoflurane at an oxygen flow rate of 1 L/min. The 508 head fur was shaved, and the incision site was sterilized with 70% ethanol prior to the beginning 509 of the surgical procedure. The mice were then mounted on a stereotaxic device (51730D, 510 Stoelting, IL), and a heat pad (BWT-100A, BRC, Nagoya) was placed underneath the mouse 511 to maintain body temperature at 37 °C. After the scalp was incised and pulled aside, a 2 mm- 512 diameter craniotomy was created with a surgical drill on the skull above the injection site. 513 Through a glass-capillary injection pipette (25 µm inner-diameter tip), 1 μl of AAV5-Syn- 514 GCaMP6f-WPRE-SV40 (titre 2.8 × 1013 GC/ml; gift from Douglas Kim & GENIE Project; 515 Addgene viral preparation #100837-AAV5)62 was injected using a microinjection pump 516 (Nanoliter 2010, WPI, FL). . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint induced not only in granule cells, but also in mossy cells. 521 522 GRIN lens implantation 523 At least two weeks after the viral injection, a gradient refractive index (GRIN) lens (Inscopix 524 1050-002202; 1 mm diameter; 4.0 mm length) was implanted into each mouse. The mouse was 525 mounted on the stereotaxic device as described above, and the scalp was removed. After 526 exposure of the skull and removal of the overlying connective tissue, we made a cranial hole 527 that was slightly larger than the diameter of the GRIN lens. To make a “pre-track” for the GRIN 528 lens insertion, we made an approximately 2 mm-wide incision in the exposed cortex down to 529 1 mm from the brain surface and then slowly embedded the lens into the dorsal DG (AP: –2.0 530 mm, ML: +1.0 mm, DV: –1.75 mm). After temporarilly immobilizing the lens with a UV- 531 curable resin (Primefil, Tokuyama Dental, Tokyo), the lens and a stainless frame (CF-10, 532 Narishige, Tokyo) were fixed to the exposed skull using dental cement mixed with black dye 533 (Sudan Black, Sigma, MO). The exposed tip of the GRIN lens was covered with dental silicone 534 (Dent Silicone-V, Shofu, Kyoto). After lens implantation, analgesic and anti-inflammatory 535 agents (Flunixin, 2 mg/kg; Fujita, Tokyo) and antibiotics (Tribrissen, 0.12 ml/kg; Kyoritsu 536 Seiyaku, Tokyo) were injected intraperitoneally. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 By monitoring the fluorescent images of GCaMP-expressing neurons, 546 the optimal location was determined (where the largest number of neurons were in focus), and 547 the baseplate was fixed with dental resin cement (Super-Bond C&B, Sun Medical, Shiga) at 548 this position. In cases where we failed to identify neurons at this stage (usually due to the failure 549 of GRIN lens implantation), the baseplate was not attached, and such mice were excluded from 550 the experiment. After baseplate attachment, a baseplate cover was placed on the baseplate until 551 Ca2+ imaging was performed. 552 553 554 555 Baseplate attachment 541 At least two weeks after GRIN lens implantation, the baseplate of a miniature micro-endoscope 542 (nVista 2.0, Inscopix, CA) was attached over the GRIN lens by a conventional method31. 543 Briefly, the mice were anaesthetized and mounted onto the stereotaxic device as described 544 above, and a baseplate attached to the miniature microscope was placed on the GRIN lens using 545 Gripper (Inscopix, CA). By monitoring the fluorescent images of GCaMP-expressing neurons, 546 the optimal location was determined (where the largest number of neurons were in focus), and 547 the baseplate was fixed with dental resin cement (Super-Bond C&B, Sun Medical, Shiga) at 548 this position. In cases where we failed to identify neurons at this stage (usually due to the failure 549 of GRIN lens implantation), the baseplate was not attached, and such mice were excluded from 550 the experiment. After baseplate attachment, a baseplate cover was placed on the baseplate until 551 Ca2+ imaging was performed. 552 553 554 555 Baseplate attachment 541 At least two weeks after GRIN lens implantation, the baseplate of a miniature micro-endoscope 542 (nVista 2.0, Inscopix, CA) was attached over the GRIN lens by a conventional method31. 543 Briefly, the mice were anaesthetized and mounted onto the stereotaxic device as described 544 above, and a baseplate attached to the miniature microscope was placed on the GRIN lens using 545 Gripper (Inscopix, CA). By monitoring the fluorescent images of GCaMP-expressing neurons, 546 the optimal location was determined (where the largest number of neurons were in focus), and 547 the baseplate was fixed with dental resin cement (Super-Bond C&B, Sun Medical, Shiga) at 548 this position. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 513 Through a glass-capillary injection pipette (25 µm inner-diameter tip), 1 μl of AAV5-Syn- 514 GCaMP6f-WPRE-SV40 (titre 2.8 × 1013 GC/ml; gift from Douglas Kim & GENIE Project; 515 Addgene viral preparation #100837-AAV5)62 was injected using a microinjection pump 516 (Nanoliter 2010, WPI, FL). After the injection, the scalp was sutured and treated with 517 povidone-iodine. DG consists of granule cells (~90%) and contains other types of excitatory 518 neurons, such as mossy cells in hilus and interneurons47. GCaMP6f expression is driven by the 519 Syn-promoter; however, it is not cell type specific. Therefore, its expression is expected to be 520 Viral delivery of Ca sensor 503 For delivery of a fluorescent Ca2+ sensor into dorsal DG neurons, adeno-associated virus (AAV) 504 carrying the GCaMP6f vector was injected into the DG of the dorsal hippocampus in adult 505 mice by a conventional method (right hemisphere; 2.0 mm posterior to bregma (AP), 1.0 mm 506 lateral to midline (ML), and 2.0 mm ventral to bregma (DV))61. In brief, adult mice >8 weeks 507 old were first anaesthetized with 1.5–3% isoflurane at an oxygen flow rate of 1 L/min. The 508 head fur was shaved, and the incision site was sterilized with 70% ethanol prior to the beginning 509 of the surgical procedure. The mice were then mounted on a stereotaxic device (51730D, 510 Stoelting, IL), and a heat pad (BWT-100A, BRC, Nagoya) was placed underneath the mouse 511 to maintain body temperature at 37 °C. After the scalp was incised and pulled aside, a 2 mm- 512 diameter craniotomy was created with a surgical drill on the skull above the injection site. 513 Through a glass-capillary injection pipette (25 µm inner-diameter tip), 1 μl of AAV5-Syn- 514 GCaMP6f-WPRE-SV40 (titre 2.8 × 1013 GC/ml; gift from Douglas Kim & GENIE Project; 515 Addgene viral preparation #100837-AAV5)62 was injected using a microinjection pump 516 (Nanoliter 2010, WPI, FL). After the injection, the scalp was sutured and treated with 517 povidone-iodine. DG consists of granule cells (~90%) and contains other types of excitatory 518 neurons, such as mossy cells in hilus and interneurons47. GCaMP6f expression is driven by the 519 Syn-promoter; however, it is not cell type specific. Therefore, its expression is expected to be 520 . CC-BY 4.0 International license perpetuity. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 After each experiment, we confirmed that the 537 GRIN lens was implanted in the upper side of the granule cell layer, where most neurons are 538 GCs, by histological analysis (Supplementary Fig. 11). 539 540 induced not only in granule cells, but also in mossy cells. 521 522 GRIN lens implantation 523 At least two weeks after the viral injection, a gradient refractive index (GRIN) lens (Inscopix 524 1050-002202; 1 mm diameter; 4.0 mm length) was implanted into each mouse. The mouse was 525 mounted on the stereotaxic device as described above, and the scalp was removed. After 526 exposure of the skull and removal of the overlying connective tissue, we made a cranial hole 527 that was slightly larger than the diameter of the GRIN lens. To make a “pre-track” for the GRIN 528 lens insertion, we made an approximately 2 mm-wide incision in the exposed cortex down to 529 1 mm from the brain surface and then slowly embedded the lens into the dorsal DG (AP: –2.0 530 mm, ML: +1.0 mm, DV: –1.75 mm). After temporarilly immobilizing the lens with a UV- 531 curable resin (Primefil, Tokuyama Dental, Tokyo), the lens and a stainless frame (CF-10, 532 Narishige, Tokyo) were fixed to the exposed skull using dental cement mixed with black dye 533 (Sudan Black, Sigma, MO). The exposed tip of the GRIN lens was covered with dental silicone 534 (Dent Silicone-V, Shofu, Kyoto). After lens implantation, analgesic and anti-inflammatory 535 agents (Flunixin, 2 mg/kg; Fujita, Tokyo) and antibiotics (Tribrissen, 0.12 ml/kg; Kyoritsu 536 Seiyaku, Tokyo) were injected intraperitoneally. After each experiment, we confirmed that the 537 GRIN lens was implanted in the upper side of the granule cell layer, where most neurons are 538 GCs, by histological analysis (Supplementary Fig. 11). 539 540 Baseplate attachment 541 At least two weeks after GRIN lens implantation, the baseplate of a miniature micro-endoscope 542 (nVista 2.0, Inscopix, CA) was attached over the GRIN lens by a conventional method31. 543 Briefly, the mice were anaesthetized and mounted onto the stereotaxic device as described 544 above, and a baseplate attached to the miniature microscope was placed on the GRIN lens using 545 Gripper (Inscopix, CA). Ca2+ imaging in freely moving mice 556 Ca2+ imaging in freely moving mice 556 Ca2+ imaging of DG neurons was performed while the mice were freely travelling in an open 557 field (OF) or T-maze, each on a different day. Each different group of mice was used for OF or 558 T-maze. Prior to Ca2+ imaging, the mice were lightly anaesthetized, and a miniature microscope 559 (nVista 2.0, Inscopix, CA) was mounted onto the baseplate of the mice. The mice were then 560 placed back in the home cages, which were then transferred to a sound-proof behavioural 561 experiment room. At least 30 min after recovery from anaesthesia, the mice were subjected to 562 OF or T-maze tests while the Ca2+ signals of their DG neurons were obtained at a 3 Hz sampling 563 rate with 1440 × 1080 pixel resolution. We applied 475/10 nm LED light for the excitation of 564 GCaMP6f fluorescence (approximately 0.24 mW/mm2 at the bottom of the GRIN lens). 565 566 567 568 Behavioural experiments 569 Before every behavioural experiment, the OF or T-maze apparatus was cleaned using weakly 570 acidified hypochlorous water (super hypochlorous water; Shimizu Laboratory Supplies, Kyoto, 571 Japan) to prevent bias due to olfactory cues. All behavioural experiments were carried out in a 572 sound-proof room, and the behaviour of the mice was monitored through a computer screen 573 located outside the room to minimize artefactual cues due to the presence of the experimenter. 574 Mouse behaviour was recorded at a 3 Hz sampling rate. 575 576 Open field test 577 More than a week after baseplate attachment, mice were habituated to the test environment. 578 Each mouse was lightly anaesthetized, and a dummy camera (Inscopix, CA) was mounted on 579 the mouse. At least 30 min after recovery from anesthesia, mice were placed for 2 hr in the OF 580 arena (40 cm × 40 cm × 30 cm; width, depth, and height, respectively; O’Hara, Japan), made 581 of opaque white plastic. The OF apparatus was evenly illuminated with 100 lux white LED 582 light installed above the apparatus. This habituation session was repeated for three days. One 583 day after the final habituation session, OF experiment was performed. Prior to experiment, the 584 mice were weakly anaesthetized with isoflurane, and an nVista miniature microscope was 585 mounted onto the head stage. 6. Information about current and future LR locations is widely distributed in the 80 population activity patterns of dDG neurons 81 In cases where we failed to identify neurons at this stage (usually due to the failure 549 of GRIN lens implantation), the baseplate was not attached, and such mice were excluded from 550 the experiment. After baseplate attachment, a baseplate cover was placed on the baseplate until 551 Ca2+ imaging was performed. 552 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Ca2+ imaging in freely moving mice 556 The mice were then habituated in the testing room for at least 30 586 min after recovery from the anaesthesia. Following the habituation in the testing room, each 587 mouse was placed in the OF arena, and neuronal activity was recorded for 30 min. In order to 588 obtain the location time sequence of each mouse, the images of the mouse were automatically 589 processed by an ImageJ plugin (Image OF, freely available on the Mouse Phenotype Database 590 Ca2+ imaging of DG neurons was performed while the mice were freely travelling in an open 557 field (OF) or T-maze, each on a different day. Each different group of mice was used for OF or 558 T-maze. Prior to Ca2+ imaging, the mice were lightly anaesthetized, and a miniature microscope 559 (nVista 2.0, Inscopix, CA) was mounted onto the baseplate of the mice. The mice were then 560 placed back in the home cages, which were then transferred to a sound-proof behavioural 561 experiment room. At least 30 min after recovery from anaesthesia, the mice were subjected to 562 OF or T-maze tests while the Ca2+ signals of their DG neurons were obtained at a 3 Hz sampling 563 rate with 1440 × 1080 pixel resolution. We applied 475/10 nm LED light for the excitation of 564 GCaMP6f fluorescence (approximately 0.24 mW/mm2 at the bottom of the GRIN lens). 565 566 p f More than a week after baseplate attachment, mice were habituated to the test environment. 578 Each mouse was lightly anaesthetized, and a dummy camera (Inscopix, CA) was mounted on 579 the mouse. At least 30 min after recovery from anesthesia, mice were placed for 2 hr in the OF 580 arena (40 cm × 40 cm × 30 cm; width, depth, and height, respectively; O’Hara, Japan), made 581 of opaque white plastic. The OF apparatus was evenly illuminated with 100 lux white LED 582 light installed above the apparatus. This habituation session was repeated for three days. One 583 day after the final habituation session, OF experiment was performed. Prior to experiment, the 584 mice were weakly anaesthetized with isoflurane, and an nVista miniature microscope was 585 mounted onto the head stage. The mice were then habituated in the testing room for at least 30 586 min after recovery from the anaesthesia. Ca2+ imaging in freely moving mice 556 Following the habituation in the testing room, each 587 mouse was placed in the OF arena, and neuronal activity was recorded for 30 min. In order to 588 obtain the location time sequence of each mouse, the images of the mouse were automatically 589 processed by an ImageJ plugin (Image OF, freely available on the Mouse Phenotype Database 590 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint website: http://www.mouse-phenotype.org/software.html). 591 592 T-maze test 593 The T-maze test was conducted using an automatic T-maze apparatus (O'Hara, Japan) as 594 previously described46. In brief, the maze consists of the stem of the T (13 cm × 24 cm); the 595 left and right (L/R) arms (11.5 cm × 20.5 cm each side); and connecting passageways from the 596 end of the L/R arms to the starting compartment. These compartments are partitioned by sliding 597 doors that open downward. The mice were subjected to a spontaneous alternation protocol for 598 five sessions, with at least 1 day (2 days maximum) of rest between sessions. Each session 599 consisted of 10 trials with a 50-min cut-off time, and each trial consisted of a first and second 600 run. On the first run, the mouse was forced to choose one of the L/R arms (forced choice). After 601 the mouse was in the L/R arm for more than 10 sec, the door to the connecting passageway 602 was opened, which allowed the mouse to return to the starting compartment. When the mouse 603 returned to the starting compartment, all the doors of this component were closed; then, after 604 3 sec, the doors connecting to the L/R arms opened for the second run, in which the mouse 605 could freely choose either of the L/R arms (free choice). Ca2+ imaging in freely moving mice 556 The percentage of trials in which the 606 mouse entered the arm opposite to their forced-choice run (the “correct” arm) was calculated. 607 The choice of the L/R arm for the forced trials was varied pseudo-randomly across trials using 608 a Gellermann schedule so that the mice received equal numbers of left and right presentations. 609 Data acquisition, control of the sliding doors, and data analysis were performed with ImageTM 610 software (freely available on the Mouse Phenotype Database website: http://www.mouse- 611 phenotype.org/software.html) 612 613 592 T-maze test 593 The T-maze test was conducted using an automatic T-maze apparatus (O'Hara, Japan) as 594 previously described46. In brief, the maze consists of the stem of the T (13 cm × 24 cm); the 595 left and right (L/R) arms (11.5 cm × 20.5 cm each side); and connecting passageways from the 596 end of the L/R arms to the starting compartment. These compartments are partitioned by sliding 597 doors that open downward. The mice were subjected to a spontaneous alternation protocol for 598 five sessions, with at least 1 day (2 days maximum) of rest between sessions. Each session 599 consisted of 10 trials with a 50-min cut-off time, and each trial consisted of a first and second 600 run. On the first run, the mouse was forced to choose one of the L/R arms (forced choice). After 601 the mouse was in the L/R arm for more than 10 sec, the door to the connecting passageway 602 was opened, which allowed the mouse to return to the starting compartment. When the mouse 603 returned to the starting compartment, all the doors of this component were closed; then, after 604 3 sec, the doors connecting to the L/R arms opened for the second run, in which the mouse 605 could freely choose either of the L/R arms (free choice). The percentage of trials in which the 606 mouse entered the arm opposite to their forced-choice run (the “correct” arm) was calculated. 607 The choice of the L/R arm for the forced trials was varied pseudo-randomly across trials using 608 a Gellermann schedule so that the mice received equal numbers of left and right presentations. Ca2+ imaging in freely moving mice 556 Finally, the temporal pattern of each the timings of DG 630 neuron’s activity was identified by an event detection algorithm available in IDPS (event 631 threshold factor, 4 median absolute deviations; event smallest decay time, 0.2 sec) and the 632 temporal pattern of activity was expressed as a time series of binarized signals for further 633 analysis. 634 cortex and CA1 of the hippocampus, were used (number of ICs, 120; number of PCs, 150; ICA 626 max iterations, 100; ICA random seed, 0; ICA convergence threshold, 0.00001; block size, 627 1000; ICA unmixing dimension, spatial; ICA temporal weights, 0.00; arbitrary units within 628 IDPS). After cell identification, longitudinal cell registration was performed to align cell maps 629 across multiple sessions as necessary. Finally, the temporal pattern of each the timings of DG 630 neuron’s activity was identified by an event detection algorithm available in IDPS (event 631 threshold factor, 4 median absolute deviations; event smallest decay time, 0.2 sec) and the 632 temporal pattern of activity was expressed as a time series of binarized signals for further 633 analysis. 634 Estimation of the active population of DG neurons 638 Estimation of the active population of DG neurons was performed on the basis of the 639 instructions of Dr. Jonathan Zapata, Inscopix Inc.. In brief, after motion correction of the raw 640 fluorescent images from the maximum intensity projection of GCaMP fluorescence images, 641 the total number of neurons in a field of view was manually counted. Likewise, the maximum 642 intensity projection of the fluorescence change ratio (ΔF/F) images was generated, and the total 643 number of active neurons during the 30 min recording was manually counted. The maximum 644 projections of GCaMP fluorescence images and ΔF/F images were calculated throughout all 645 frames in the 30 min recording in the OF using the “movie projection” tool available in IDPS 646 and saved as tiff files. We counted the number of cells in the maximum projection image frames 647 using the multipoint tool of ImageJ (available from https://imagej.nih.gov/ij/). To confirm the 648 activity of all the detected cells, we manually drew regions of interest (ROIs) over all the cells 649 in the ΔF/F projection images and confirmed the traces of the Ca2+ signals with IDPS. All 650 neurons that exhibited calcium transients individually in the soma at least once during the entire 651 recording period were considered “active”. Ca2+ imaging in freely moving mice 556 634 635 636 637 Estimation of the active population of DG neurons 638 Estimation of the active population of DG neurons was performed on the basis of the 639 instructions of Dr. Jonathan Zapata, Inscopix Inc.. In brief, after motion correction of the raw 640 fluorescent images from the maximum intensity projection of GCaMP fluorescence images, 641 the total number of neurons in a field of view was manually counted. Likewise, the maximum 642 intensity projection of the fluorescence change ratio (ΔF/F) images was generated, and the total 643 number of active neurons during the 30 min recording was manually counted. The maximum 644 projections of GCaMP fluorescence images and ΔF/F images were calculated throughout all 645 frames in the 30 min recording in the OF using the “movie projection” tool available in IDPS 646 and saved as tiff files. We counted the number of cells in the maximum projection image frames 647 using the multipoint tool of ImageJ (available from https://imagej.nih.gov/ij/). To confirm the 648 activity of all the detected cells, we manually drew regions of interest (ROIs) over all the cells 649 in the ΔF/F projection images and confirmed the traces of the Ca2+ signals with IDPS. All 650 neurons that exhibited calcium transients individually in the soma at least once during the entire 651 recording period were considered “active”. All the image data that were used to perform active 652 cell counting and the detailed method are available at “SSBD:repository” database 653 [http://ssbd.qbic.riken.jp/set/20200603/]. 654 655 656 Definitions of the position, speed, and motion direction of mice 657 (i) Position: The OF arena (which had an area of 40 cm × 40 cm) was represented as 200 × 200 658 cortex and CA1 of the hippocampus, were used (number of ICs, 120; number of PCs, 150; ICA 626 max iterations, 100; ICA random seed, 0; ICA convergence threshold, 0.00001; block size, 627 1000; ICA unmixing dimension, spatial; ICA temporal weights, 0.00; arbitrary units within 628 IDPS). After cell identification, longitudinal cell registration was performed to align cell maps 629 across multiple sessions as necessary. Ca2+ imaging in freely moving mice 556 609 Data acquisition, control of the sliding doors, and data analysis were performed with ImageTM 610 software (freely available on the Mouse Phenotype Database website: http://www.mouse- 611 phenotype.org/software.html) 612 613 614 615 Detection of Ca2+ transients 616 To extract the activity patterns of individual DG neurons from the obtained fluorescent images, 617 we used Inscopix Data Processing Software (IDPS). Briefly, the data files of the raw sequential 618 fluorescent images obtained by nVista (Inscopix) were imported to IDPS, and motion 619 correction was applied. The instantaneous fluorescence of the motion-corrected images was 620 then normalized by its average fluorescence over the entire recording period, producing 621 fluorescence change ratio (ΔF/F) images. Then, individual DG neurons were identified by 622 automated PCA/ICA (PCA, principal component analysis; ICA, independent component 623 analysis) segmentation of activity traces. For PCA/ICA segmentation, default parameters, 624 whose values were confirmed to work well across most of the neuronal activity patterns in the 625 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint cortex and CA1 of the hippocampus, were used (number of ICs, 120; number of PCs, 150; ICA 626 max iterations, 100; ICA random seed, 0; ICA convergence threshold, 0.00001; block size 627 1000; ICA unmixing dimension, spatial; ICA temporal weights, 0.00; arbitrary units within 628 IDPS). After cell identification, longitudinal cell registration was performed to align cell maps 629 across multiple sessions as necessary. Finally, the temporal pattern of each the timings of DG 630 neuron’s activity was identified by an event detection algorithm available in IDPS (even 631 threshold factor, 4 median absolute deviations; event smallest decay time, 0.2 sec) and the 632 temporal pattern of activity was expressed as a time series of binarized signals for further 633 analysis. Ca2+ imaging in freely moving mice 556 634 635 636 637 Estimation of the active population of DG neurons 638 Estimation of the active population of DG neurons was performed on the basis of the 639 instructions of Dr. Jonathan Zapata, Inscopix Inc.. In brief, after motion correction of the raw 640 fluorescent images from the maximum intensity projection of GCaMP fluorescence images 641 the total number of neurons in a field of view was manually counted. Likewise, the maximum 642 intensity projection of the fluorescence change ratio (ΔF/F) images was generated, and the tota 643 number of active neurons during the 30 min recording was manually counted. The maximum 644 projections of GCaMP fluorescence images and ΔF/F images were calculated throughout al 645 frames in the 30 min recording in the OF using the “movie projection” tool available in IDPS 646 and saved as tiff files. We counted the number of cells in the maximum projection image frames 647 using the multipoint tool of ImageJ (available from https://imagej.nih.gov/ij/). To confirm the 648 activity of all the detected cells, we manually drew regions of interest (ROIs) over all the cells 649 in the ΔF/F projection images and confirmed the traces of the Ca2+ signals with IDPS. Al 650 neurons that exhibited calcium transients individually in the soma at least once during the entire 651 recording period were considered “active”. All the image data that were used to perform active 652 cell counting and the detailed method are available at “SSBD:repository” database 653 [http://ssbd.qbic.riken.jp/set/20200603/]. 654 655 656 Definitions of the position, speed, and motion direction of mice 657 (i) P i i Th OF ( hi h h d f 40 40 ) d 200 200 658 cortex and CA1 of the hippocampus, were used (number of ICs, 120; number of PCs, 150; ICA 626 max iterations, 100; ICA random seed, 0; ICA convergence threshold, 0.00001; block size, 627 1000; ICA unmixing dimension, spatial; ICA temporal weights, 0.00; arbitrary units within 628 IDPS). After cell identification, longitudinal cell registration was performed to align cell maps 629 across multiple sessions as necessary. Finally, the temporal pattern of each the timings of DG 630 neuron’s activity was identified by an event detection algorithm available in IDPS (event 631 threshold factor, 4 median absolute deviations; event smallest decay time, 0.2 sec) and the 632 temporal pattern of activity was expressed as a time series of binarized signals for further 633 analysis. Ca2+ imaging in freely moving mice 556 All the image data that were used to perform active 652 cell counting and the detailed method are available at “SSBD:repository” database 653 [http://ssbd.qbic.riken.jp/set/20200603/]. 654 655 f f p p f (i) Position: The OF arena (which had an area of 40 cm × 40 cm) was represented as 200 × 200 658 pixel grid. The position of the mouse is determined from the centroid of its shadow on the 659 camera. We then assigned a label corresponding to the discrete location of the mouse (e.g., [10, 660 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 100]) to each time bin (=1/3 sec). 661 (ii) Speed: From the distance travelled between 1 sec before and after a given time point, we 662 calculated the speed of mouse at that moment, and assigned this speed (cm/sec) to each time 663 bin. 664 (iii) Motion direction: The visual tracking system that we used does not allow direct 665 measurement of head direction. Instead, we indirectly estimated the direction of motion from 666 the changes in the position of the mouse. The motion direction (in radians) was computed from 667 the direction of change in two subsequent mouse positions (1 sec before and after a given time 668 point) in the x-y plane and assigned this to each time bin. North was defined as 0 radians; west 669 was defined from 0 to π radians, and east was defined from 0 to –π radians. 670 671 672 673 Statistical analysis of spatial, speed and direction information 674 To quantify the tuning specificities of neurons with position, speed, and motion direction, we 675 measured their specificity in terms of the information rate of cell activity and defined them as 676 (i) spatial, (ii) speed, and (iii) direction information44. Ca2+ imaging in freely moving mice 556 721 Random permutation permutates calcium events (Supplementary Figure 12). We divided the 722 calcium event data into 1000 segments along the time axis and randomly sorted them to 723 generate permutated data of calcium events. This method destroys temporal structures of neural 724 activity and temporal correlations between neural activity and behavioral variables (e.g., 725 position, speed, and motion direction in open field test); however, the overall neural activity is 726 maintained across cells We repeated this procedure 1000 times to obtain the distribution of 727 transient rate. 694 (ii) To measure speed information, we applied the same form 695 it to a binary state: Run (for speeds >1 cm/sec) and Stop (fo 696 (iii) Similarly, we measured direction information by applyi 697 direction after discretizing the full angle to 8 bins of 45 deg 698 699 700 701 LR indices of neurons in the T-maze test 702 To quantify the left- or right- preference of each neuron’s 703 location in the T-maze, we generated a parameter called th 704 measured the average Ca2+ transient rates during the forced 705 left- and right-choice trials—that is, when the mouse is in t 706 respectively—and computed the LR index as follows: 707 708 709 710 where RL is the average Ca2+ transient rate across all left-ch 711 right-choice trials. All neurons are assigned an LR index in 712 periods. A positive value indicates a neuron’s preference fo 713 value indicates a neuron’s preference for the right arm. 714 715 716 717 Data Shuffling 718 To assess the statistical significance of information coding o 719 chance distributions of the shuffled data using two commo 720 the literature32,18,9. 721 Random permutation permutates calcium events (Supplem 722 calcium event data into 1000 segments along the time a 723 generate permutated data of calcium events. This method de 724 activity and temporal correlations between neural activit 725 position, speed, and motion direction in open field test); how 726 maintained across cells We repeated this procedure 1000 727 To quantify the left- or right- preference of each neuron’s activity for the current and future 703 location in the T-maze, we generated a parameter called the LR index. Ca2+ imaging in freely moving mice 556 It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for thi this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint transient rate. 694 (ii) To measure speed information, we applied the same formula to the speed after discretizing 695 it to a binary state: Run (for speeds >1 cm/sec) and Stop (for speeds <1 cm/sec). 696 (iii) Similarly, we measured direction information by applying the same formula to the motion 697 direction after discretizing the full angle to 8 bins of 45 degrees each. 698 699 700 701 LR indices of neurons in the T-maze test 702 To quantify the left- or right- preference of each neuron’s activity for the current and future 703 location in the T-maze, we generated a parameter called the LR index. For each neuron, we 704 measured the average Ca2+ transient rates during the forced arm and decision periods for the 705 left- and right-choice trials—that is, when the mouse is in the left or right arm of the T-maze, 706 respectively—and computed the LR index as follows: 707 708 709 710 where RL is the average Ca2+ transient rate across all left-choice trials, and RR is that from the 711 right-choice trials. All neurons are assigned an LR index in both the forced arm and decision 712 periods. A positive value indicates a neuron’s preference for the left arm, whereas a negative 713 value indicates a neuron’s preference for the right arm. 714 715 716 717 Data Shuffling 718 To assess the statistical significance of information coding of individual neurons, we computed 719 chance distributions of the shuffled data using two common methods previously described in 720 the literature32,18,9. Ca2+ imaging in freely moving mice 556 The Ca2+ event rate in Ca2+ imaging is 677 considerably lower than in electrophysiological recordings (on average, approximately 30 Ca2+ 678 transients per 30 min session). If the discretization of position, speed, and motion direction it 679 too fine for the number of events in the recorded cells, we will not be able to obtain a proper 680 null distribution when creating the shuffle data for that cell. Therefore, we set the resolution of 681 the discretization of position and speed to be lower than those commonly performed. 682 (i) We used a 2×2 square grid for measurement of spatial information and computed the amount 683 of Shannon information that a single Ca2+ transient conveyed about the animal’s position. The 684 spatial information I (bits per Ca2+ transient) of a cell was calculated as the mutual information 685 score between the occurrence of a single Ca2+ transient of the cell and the animal’s behavioural 686 state of position using the formula: 687 688 (i) We used a 2×2 square grid for measurement of spatial information and computed the amount 683 of Shannon information that a single Ca2+ transient conveyed about the animal’s position. The 684 spatial information I (bits per Ca2+ transient) of a cell was calculated as the mutual information 685 score between the occurrence of a single Ca2+ transient of the cell and the animal’s behavioural 686 state of position using the formula: 687 689 where i is the bin number corresponding to the physical parameter (in this case, spatial position 691 in the OF; i = 1–4 from the 2×2 square grid), N is the total number of bins, p is the probability 692 that the mouse occupied bin i, ri is the mean transient rate at bin i, and r is the overall mean 693 where i is the bin number corresponding to the physical parameter (in this case, spatial position 691 in the OF; i = 1–4 from the 2×2 square grid), N is the total number of bins, p is the probability 692 that the mouse occupied bin i, ri is the mean transient rate at bin i, and r is the overall mean 693 . CC-BY 4.0 International license perpetuity. Ca2+ imaging in freely moving mice 556 This procedure disrupts the relationship 739 between neural activity and animal behaviour, but preserves the temporal patterns of these 740 variables. By repeating this procedure while changing the number of frames to be shifted at 741 random, we obtain the null distributions of shuffled data (Supplementary Figure 12). 742 Information statistics are performed in the same way as the first shuffling method. 743 Since we found that the information statistics and decoding results of the shuffled data did not 744 differ significantly between these shuffling strategies (Supplementary Figure 12), we adopted 745 the random permutation method for the generation of shuffled data. 746 747 748 749 Decoding position, speed, and motion direction in the open field 750 To determine the manner in which the OF behavioural parameters are encoded in the DG, we 751 trained decoders with machine learning methods to separately predict position, speed, and 752 motion direction from the population Ca2+ activity. We assigned the labels of the discretized 753 behavioural parameters of the mouse (position (cm), speed (cm/sec), and motion direction (–π 754 – +π radians)) and the binary values of the Ca2+ signal (0 or 1) of all neurons to each time bin. 755 We then divided the Ca2+ imaging data and behavioural data from each 30 min trial into the 756 first 15 min and last 15 min halves, which were designated training and test data, respectively. 757 For each pair of behavioural parameter (position, speed, or motion direction) and the value of 758 the Ca2+ signal in the training data, we trained the decoders with one of eight different machine 759 learning methods (Dense Feedforward Neural Network (DNN), Gated Recurrent Unit (GRU), 760 Long Short-Term Memory (LSTM) network, Recurrent Neural Network (RNN), Support 761 Vector Regression (SVR), Wiener Cascade (WC), Wiener Filter (WF), Extreme Gradient 762 1000 shuffled data. For single-cell statistics, we compared the original information of 728 individual cells with the null distributions of 1000 values of shuffled data generated from the 729 original cell (Supplementary Figure 2). If the original value of information of a cell exceeded 730 3 sigma from the shuffled distribution, the cells were defined as carrying significant amounts 731 of information. For group comparison (for example, Obs. vs Shuff. in Fig. Ca2+ imaging in freely moving mice 556 For each neuron, we 704 measured the average Ca2+ transient rates during the forced arm and decision periods for the 705 left- and right-choice trials—that is, when the mouse is in the left or right arm of the T-maze, 706 respectively—and computed the LR index as follows: 707 where RL is the average Ca2+ transient rate across all left-choice trials, and RR is that from the 711 right-choice trials. All neurons are assigned an LR index in both the forced arm and decision 712 periods. A positive value indicates a neuron’s preference for the left arm, whereas a negative 713 value indicates a neuron’s preference for the right arm. 714 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 1000 shuffled data. For single-cell statistics, we compared the original information of 728 individual cells with the null distributions of 1000 values of shuffled data generated from the 729 original cell (Supplementary Figure 2). If the original value of information of a cell exceeded 730 3 sigma from the shuffled distribution, the cells were defined as carrying significant amounts 731 of information. For group comparison (for example, Obs. vs Shuff. in Fig. 1d, 4c, and 4d), we 732 pooled all the shuffled data in a group together, and distributions of the original cells were 733 compared with null distributions of the shuffled data. 734 Random scrambling is a method that maintains temporal dynamics of neural activity data while 735 disrupting the relationship with behavioral patterns, (for example, the calcium event timeseries 736 and the animal’s position (Supplementary Figure 12)). We shift the whole vector of the calcium 737 event time series in time by a random amount in a torus, that is, points that went beyond the 738 time limits of the data were reinsert from the other side. Ca2+ imaging in freely moving mice 556 1d, 4c, and 4d), we 732 pooled all the shuffled data in a group together, and distributions of the original cells were 733 compared with null distributions of the shuffled data. 734 1000 shuffled data. For single-cell statistics, we compared the original information of 728 individual cells with the null distributions of 1000 values of shuffled data generated from the 729 original cell (Supplementary Figure 2). If the original value of information of a cell exceeded 730 3 sigma from the shuffled distribution, the cells were defined as carrying significant amounts 731 of information. For group comparison (for example, Obs. vs Shuff. in Fig. 1d, 4c, and 4d), we 732 pooled all the shuffled data in a group together, and distributions of the original cells were 733 compared with null distributions of the shuffled data. 734 Random scrambling is a method that maintains temporal dynamics of neural activity data while 735 disrupting the relationship with behavioral patterns, (for example, the calcium event timeseries 736 and the animal’s position (Supplementary Figure 12)). We shift the whole vector of the calcium 737 event time series in time by a random amount in a torus, that is, points that went beyond the 738 time limits of the data were reinsert from the other side. This procedure disrupts the relationship 739 between neural activity and animal behaviour, but preserves the temporal patterns of these 740 variables. By repeating this procedure while changing the number of frames to be shifted at 741 random, we obtain the null distributions of shuffled data (Supplementary Figure 12). 742 Information statistics are performed in the same way as the first shuffling method. 743 Decoding position, speed, and motion direction in the open field 750 To determine the manner in which the OF behavioural parameters are encoded in the DG, we 751 trained decoders with machine learning methods to separately predict position, speed, and 752 motion direction from the population Ca2+ activity. We assigned the labels of the discretized 753 behavioural parameters of the mouse (position (cm), speed (cm/sec), and motion direction (–π 754 – +π radians)) and the binary values of the Ca2+ signal (0 or 1) of all neurons to each time bin. 755 We then divided the Ca2+ imaging data and behavioural data from each 30 min trial into the 756 first 15 min and last 15 min halves, which were designated training and test data, respectively. Ca2+ imaging in freely moving mice 556 For 776 decoding position and motion direction, we identified and removed time bins when the mouse 777 moved at speeds below 1.0 cm/sec and 4.0 cm/sec, respectively, to obtain optimal decoding 778 results. Details on the methods used in thresholding the data according to the movement speeds 779 of the mice are described in the Supplementary Notes and Supplementary Fig. 10. 780 781 782 783 Decoding current and future location in the T-maze test 784 We similarly sought to decode the left or right preferences for the current and future locations 785 in the T-Maze test using the population Ca2+ activity with machine learning methods. For each 786 trial of the forced arm and decision periods, we assigned a label corresponding to the left or 787 right choice of the mouse and the average Ca2+ transient rate of each neuron during the period. 788 We then used a support vector machine (SVM) classifier function in MATLAB for binary 789 classification of the left or right decision (MathWorks, MA). All 50 trials from each mouse 790 were used for 25-fold cross-validation; the 50 trials were randomly divided into 48 trials of 791 training data and 2 trials of test data. We repeatedly trained the binary classifier using randomly 792 selected training data and performed left or right predictions for the remaining test data to 793 evaluate the decoding accuracy. We also shuffled the Ca2+ transient data and performed the 794 same decoding analysis. As with the open field behavioural parameters, the decoding accuracy 795 was compared between the shuffled and unshuffled data. 796 797 Boosting (XGB); the codes were obtained and modified from Glaser et al., arXiv, 201863). The 763 decoding accuracy of the position/motion direction is reported as the mean absolute error in 764 the distance between the predicted and actual position/motion direction. In the case of speed, 765 decoding accuracy was reported as the correlation between the predicted and actual 766 instantaneous speed. This is because speed, unlike position and motion direction, is not limited 767 to a certain range and mean absolute error of speed may depends on the average locomotion 768 speed of each individual mouse, which would be inappropriate for evaluation of decoding error. Ca2+ imaging in freely moving mice 556 757 For each pair of behavioural parameter (position, speed, or motion direction) and the value of 758 the Ca2+ signal in the training data, we trained the decoders with one of eight different machine 759 learning methods (Dense Feedforward Neural Network (DNN), Gated Recurrent Unit (GRU), 760 Long Short-Term Memory (LSTM) network, Recurrent Neural Network (RNN), Support 761 Vector Regression (SVR), Wiener Cascade (WC), Wiener Filter (WF), Extreme Gradient 762 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Boosting (XGB); the codes were obtained and modified from Glaser et al., arXiv, 201863). The 763 decoding accuracy of the position/motion direction is reported as the mean absolute error in 764 the distance between the predicted and actual position/motion direction. In the case of speed, 765 decoding accuracy was reported as the correlation between the predicted and actual 766 instantaneous speed. This is because speed, unlike position and motion direction, is not limited 767 to a certain range and mean absolute error of speed may depends on the average locomotion 768 speed of each individual mouse, which would be inappropriate for evaluation of decoding error. 769 To assess the statistical significance of the decoding accuracies, the decoding error from the 770 observed data was compared with that of the shuffled data, which is created by dividing the 771 Ca2+ imaging data into 1,000 segments and sorting them randomly as described above. 772 Furthermore, we compared the decoding accuracies of the three behavioural parameters among 773 the 8 decoders and found that they were not significantly different (Supplementary Fig. 3). 774 Consequently, we reported the decoding results that were obtained with the LSTM Network, 775 which showed slightly better decoding performance in wild-type mice than others. Ca2+ imaging in freely moving mice 556 769 To assess the statistical significance of the decoding accuracies, the decoding error from the 770 observed data was compared with that of the shuffled data, which is created by dividing the 771 Ca2+ imaging data into 1,000 segments and sorting them randomly as described above. 772 Furthermore, we compared the decoding accuracies of the three behavioural parameters among 773 the 8 decoders and found that they were not significantly different (Supplementary Fig. 3). 774 Consequently, we reported the decoding results that were obtained with the LSTM Network, 775 which showed slightly better decoding performance in wild-type mice than others. For 776 decoding position and motion direction, we identified and removed time bins when the mouse 777 moved at speeds below 1.0 cm/sec and 4.0 cm/sec, respectively, to obtain optimal decoding 778 results. Details on the methods used in thresholding the data according to the movement speeds 779 of the mice are described in the Supplementary Notes and Supplementary Fig. 10. 780 781 Decoding current and future location in the T-maze test 784 We similarly sought to decode the left or right preferences for the current and future locations 785 in the T-Maze test using the population Ca2+ activity with machine learning methods. For each 786 trial of the forced arm and decision periods, we assigned a label corresponding to the left or 787 right choice of the mouse and the average Ca2+ transient rate of each neuron during the period. 788 We then used a support vector machine (SVM) classifier function in MATLAB for binary 789 classification of the left or right decision (MathWorks, MA). All 50 trials from each mouse 790 were used for 25-fold cross-validation; the 50 trials were randomly divided into 48 trials of 791 training data and 2 trials of test data. We repeatedly trained the binary classifier using randomly 792 selected training data and performed left or right predictions for the remaining test data to 793 evaluate the decoding accuracy. We also shuffled the Ca2+ transient data and performed the 794 same decoding analysis. As with the open field behavioural parameters, the decoding accuracy 795 was compared between the shuffled and unshuffled data. 796 797 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint To quantify the similarities in population activity patterns of neurons between different 801 locations in the DG, we calculated the population vector overlap (PVO)64. The PVO for a 802 population of N neurons in two different conditions (x, y) was defined as 803 804 805 805 806 where i is the neuron number; λj(x) and λj(y) are the average Ca2+ transient rates of neuron j in 807 conditions x and y, respectively; and N is the total number of neurons. For a given population 808 of neurons and a pair of conditions, a lower PVO indicates that the activity patterns of the 809 neurons are distinct between the two conditions. . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Decoding current and future location in the T-maze test 784 810 811 812 813 Code and data availability 814 The data analysis was performed using custom code written in MATLAB (MathWorks, 815 R2018a) and Python (Version 2.7.12). For decoding analysis by machine learning, we obtained 816 Python code available at https://github.com/KordingLab/Neural_Decoding (used in Glaser et 817 al., arXiv, 201863) and modified them for our purpose. Raw imaging data in this study is 818 uploaded at Systems Science Biological Dynamics repository (SSBD:repository; 819 http://ssbd.qbic.riken.jp/set/20200603). Processed imaging data and behavioural data in this 820 study are available at https://github.com/tmurano. Codes will be made available upon 821 reasonable request. 822 823 824 where i is the neuron number; λj(x) and λj(y) are the average Ca2+ transient rates of neuron j in 807 conditions x and y, respectively; and N is the total number of neurons. For a given population 808 of neurons and a pair of conditions, a lower PVO indicates that the activity patterns of the 809 neurons are distinct between the two conditions. 810 The data analysis was performed using custom code written in MATLAB (MathWorks, 815 R2018a) and Python (Version 2.7.12). For decoding analysis by machine learning, we obtained 816 Python code available at https://github.com/KordingLab/Neural_Decoding (used in Glaser et 817 al., arXiv, 201863) and modified them for our purpose. Raw imaging data in this study is 818 uploaded at Systems Science Biological Dynamics repository (SSBD:repository; 819 http://ssbd.qbic.riken.jp/set/20200603). Processed imaging data and behavioural data in this 820 study are available at https://github.com/tmurano. Codes will be made available upon 821 reasonable request. 822 825 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Supplementary Notes 826 827 828 Velocity filtering for position and direction decoding (Supplementary Fig. 10) 829 The activation of hippocampal place cells during immobile periods, which occurs in 830 conjunction with hippocampal sharp wave ripples (SWRs), is known not to represent the 831 current position of animals65,66. Decoding current and future location in the T-maze test 784 Therefore, in analyses of position information in the 832 hippocampus, periods of immobility are generally excluded from the data. We did not know 833 whether immobile periods should also be excluded for data obtained from the dDG, as it is not 834 known whether neural activity equivalent to SWRs can be observed in the Ca2+ imaging in this 835 region. Therefore, to test the validity of this procedure, we examined whether removing periods 836 of immobility would improve the accuracy of position decoding in the dDG (Supplementary 837 Fig. 10, left panel). We removed the time bins in which the movement speed of the mouse was 838 below the threshold (from 0.0 cm/sec to 3.0 cm/sec) and performed position decoding using 839 the remaining data. In wild-type mice, removing time bins in which the speed was below 1.0– 840 1.25 cm/sec reduced the position decoding error (Supplementary Fig. 10, left panel). These 841 results suggest that neural activity during periods of immobility that do not represent current 842 location may also be observed in the Ca2+ imaging data of the dDG. Based on these results, all 843 periods in which the mouse’s speed was below 1 cm/sec were eliminated from the analysis of 844 position information. (This threshold is roughly equivalent to those commonly used in 845 hippocampal CA cells (1.0–2.0 cm/sec)32). 846 Velocity filtering for position and direction decoding (Supplementary Fig. 10) The motion direction of the mouse was estimated from the changes in the mouse 847 position in the open field. Even small movements such as grooming, rearing or head turning 848 were calculated as actual mouse movements. To exclude these periods and more accurately 849 measure motion direction, we examined the speed threshold below which movements could to 850 be removed from the datasets (Supplementary Fig. 10, right panel). We removed all the time 851 bins in which the speed of the mouse was below the threshold (from 0.0 cm/sec to 10.0 cm/sec) 852 and performed direction decoding using the remaining data. In wild-type mice, the decoding 853 error for motion direction decreased after applying a threshold of 4.0–8.0 cm/sec to remove 854 movement periods, suggesting that small movement periods below these thresholds were 855 useless for accurate direction decoding (Supplementary Fig. 10, right panel). Based on these 856 results, all periods when the mouse ran slower than 4.0 cm/sec were removed from our datasets 857 for the analysis of motion direction. 858 859 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . 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Buzsaki, G., Horvath, Z., Urioste, R., Hetke, J. & Wise, K. High- 1016 frequency network oscillation in the hippocampus. Science 256, 1025–1027 (1992). 1017 66. Buzsáki, G. Hippocampal sharp wave-ripple: A cognitive biomarker for 1018 episodic memory and planning. Hippocampus 25, 1073–1188 (2015). 1019 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. Acknowledgements 1022 Acknowledgements 1022 We thank Wakako Hasegawa, Yumiko Mobayashi, Misako Murai, Tamaki Murakami, Miwa 1023 Takeuchi, Yoko Kagami, Harumi Mitsuya, Yoshihiro Takamiya, and other members of the 1024 Miyakawa laboratory for their support. We also thank Jonathan Zapata (Inscopix) for his 1025 detailed advices on the image analysis. This work was supported by a JSPS Grant-in-Aid for 1026 Scientific Research on Innovative Areas (grant #JP16H06462), the AMED Strategic Research 1027 Program for Brain Science (grant #JP18dm0107101), and a grant from Astellas Pharma Inc. 1028 1029 We thank Wakako Hasegawa, Yumiko Mobayashi, Misako Murai, Tamaki Murakami, Miwa 1023 Takeuchi, Yoko Kagami, Harumi Mitsuya, Yoshihiro Takamiya, and other members of the 1024 Miyakawa laboratory for their support. We also thank Jonathan Zapata (Inscopix) for his 1025 detailed advices on the image analysis. This work was supported by a JSPS Grant-in-Aid for 1026 Scientific Research on Innovative Areas (grant #JP16H06462), the AMED Strategic Research 1027 Program for Brain Science (grant #JP18dm0107101), and a grant from Astellas Pharma Inc. 1028 1029 33. Vincent, M. et al. Large deletions encompassing the TCOF1 and It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Acknowledgements 1022 We thank Wakako Hasegawa, Yumiko Mobayashi, Misako Murai, Tamaki Murakami, Miwa 1023 Takeuchi, Yoko Kagami, Harumi Mitsuya, Yoshihiro Takamiya, and other members of the 1024 Miyakawa laboratory for their support. We also thank Jonathan Zapata (Inscopix) for his 1025 detailed advices on the image analysis. This work was supported by a JSPS Grant-in-Aid for 1026 Scientific Research on Innovative Areas (grant #JP16H06462), the AMED Strategic Research 1027 Program for Brain Science (grant #JP18dm0107101), and a grant from Astellas Pharma Inc. 1028 1029 Author Contribution Statement 1030 T. Murano A. Nakao, R. Nakajima, J. Yamamoto, and T. Miyakawa designed the experiments 1031 and analyses. A. Nakao, N. Hirata, and R. Nakajima performed the behavioural testing and 1032 Ca2+ imaging. T. Murano, R. Nakajima, S. Amemori, A. Murakami, and Y. Kamitani analysed 1033 the data. T. Murano, R. Nakajima, J. Yamamoto, and T. Miyakawa prepared figures and wrote 1034 the manuscript. T. Miyakawa supervised all aspects of the present study. All authors reviewed 1035 the manuscript. 1036 1037 Competing Financial Interests 1038 The authors declare no competing financial interests. 1039 1040 Additional Information 1041 Processed imaging and behavioural data are available at https://github.com/tmurano. Raw 1042 images and behavioural data are available at http://ssbd.qbic.riken.jp/set/20200603/. 1043 Correspondence and requests for materials should be addressed to T.M. 1044 1045 Author Contribution Statement 1030 Author Contribution Statement 1030 T. Murano A. Nakao, R. Nakajima, J. Yamamoto, and T. Miyakawa designed the experiments 1031 and analyses. A. Nakao, N. Hirata, and R. Nakajima performed the behavioural testing and 1032 Ca2+ imaging. T. Murano, R. Nakajima, S. Amemori, A. Murakami, and Y. Kamitani analysed 1033 the data. T. Murano, R. Nakajima, J. Yamamoto, and T. Miyakawa prepared figures and wrote 1034 the manuscript. T. Miyakawa supervised all aspects of the present study. All authors reviewed 1035 the manuscript. 1036 T. Murano A. Nakao, R. Nakajima, J. Yamamoto, and T. Miyakawa designed the experiments 1031 and analyses. A. Nakao, N. Hirata, and R. Nakajima performed the behavioural testing and 1032 Ca2+ imaging. T. Murano, R. Nakajima, S. Amemori, A. Murakami, and Y. Kamitani analysed 1033 the data. T. Murano, R. Nakajima, J. Yamamoto, and T. Miyakawa prepared figures and wrote 1034 the manuscript. T. Miyakawa supervised all aspects of the present study. All authors reviewed 1035 the manuscript. 1036 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 1046 1047 Figures 1047 Figure 1. Activity patterns of dorsal DG neurons are slightly tuned to the position and speed of the mouse. 1048 Figure 1. Activity patterns of dorsal DG neurons are slightly tuned to the position and 1049 speed of the mouse. 1050 048 Figure 1. Activity patterns of dorsal DG neurons are slightly tuned to the position and 049 speed of the mouse. 050 1048 Figure 1. Activity patterns of dorsal DG neurons are slightly tuned to the position and 1049 speed of the mouse. 1050 1051 a, Schematic of the experimental procedures. GCaMP6f was expressed in the dorsal DG (dDG) 1052 of mice by AAV injection. At least 2 weeks after viral injection, a graded-index (GRIN) lens 1053 was implanted. Author Contribution Statement 1030 Two to three weeks after lens implantation, a baseplate for a miniature 1054 microscope was attached at the optimal focal plane. Over 1 week after baseplate attachment, 1055 the mice were habituated to the test environment and subjected to the behavioural experiments. 1056 1057 b, Left: Representative image of active dDG neurons in a wild-type mouse. The image shows 1058 the projection of the maximum relative fluorescence change (ΔF/F) of the Ca2+ transient among 1059 Figure 1. Activity patterns of dorsal DG neurons are slightly tuned to the position and 1049 speed of the mouse. 1050 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint the entire set of frames from the recording of the 30 min open field experiment. Orange circles 1060 indicate the regions of interest of the recorded dDG neurons. Right: ΔF/F of the Ca2+ signals 1061 for 15 cells. Scale bars: 30 sec (horizontal) and 5% ΔF/F (vertical). 1062 1063 c, Colour-coded spatial tuning maps, speed tuning curves, and direction tuning maps of three 1064 representative dDG neurons from wild-type mice (left) and αCaMKII+/– mice (right) are shown. 1065 Top: Spatial tuning maps showing the density of mouse locations where Ca2+ transients were 1066 detected. Each map is normalized to each neuron’s maximum Ca2+ transient rate and smoothed 1067 with a Gaussian kernel (σ = 2.0 cm). Middle: Speed tuning curves representing the mean Ca2+ 1068 transient rate (s–1) when the mouse moved at 0–1 cm/sec, 1–5 cm/sec, and >5 cm/sec. Bottom: 1069 Direction tuning maps showing the mean Ca2+ transient rate when the motion direction of the 1070 mouse was in each of the eight indicated directions. Each map is normalized to the neuron’s 1071 maximum mean Ca2+ transient rate. North is defined as 0 radians; west is defined from 0 to π 1072 radians, and east is defined from 0 to –π radians. Author Contribution Statement 1030 1073 1074 d, Distributions of information content from observed neurons (“Obs.”, colour-coded curves) 1075 and those from shuffled data (“Shuff.”, grey curves; 1,000 shuffles per cell; counts normalized 1076 by number of shuffles) for all neurons from all animals (508 neurons from 7 wild-type and 289 1077 neurons from 5 αCaMKII+/– mice). The dashed line shows the 95th percentile of the distribution 1078 of the shuffled data, and the numbers on the lines represent the percentage of observed neurons 1079 exceeding this value. 1080 1081 the entire set of frames from the recording of the 30 min open field experiment. Orange circles 1060 indicate the regions of interest of the recorded dDG neurons. Right: ΔF/F of the Ca2+ signals 1061 for 15 cells. Scale bars: 30 sec (horizontal) and 5% ΔF/F (vertical). 1062 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 082 Figure 2. Information about position, speed, and motion direction is encoded in the 083 population activity patterns of dDG neurons. 084 Observed Decoded -0.3 -0.1 0.1 0.3 0.5 0.7 -0.3 -0.1 0.1 0.3 0.5 0.7 1.2 1.3 1.4 1.5 1.6 1.7 1.2 1.3 1.4 1.5 1.6 1.7 a X-position (cm) Y-position (cm) Speed (cm/sec) Wild-type αCaMKII+/– Correlation Coefficient Mean Absolute Error (cm) Mean Absolute Error (radian) Speed Decoding Position Decoding Motion Direction (radian) Direction Decoding Obs. Shuff. P = 0.262 c b Time (sec) Time (sec) Time (sec) Time (sec) Time (sec) Time (sec) Observed Decoded Obs. Shuff. Obs. Shuff. Obs. Shuff. Obs. Shuff. Obs. Shuff. P = 0.619 P = 1.1210–3 () d Speed Decoding Position Decoding Direction Decoding f e Wild-type αCaMKII+/– Observed Decoded 10 14 18 22 10 14 18 22 a X-position (cm) Y-position (cm) Wild-type αCaMKII+/– Position Decoding b Time (sec) Time (sec) Observed Decoded Mean Absolute Error (cm) Obs. Shuff. Obs. Shuff. Author Contribution Statement 1030 It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint a Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) Wild-type Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) R = 0.167 P = 1.5710–4 R = 0.257 P = 3.9510–9 R = –1.6710–4 P = 0.997 2% 3% 2% 3% 29% 7% 9% 24% 14% 60% 66% 81% a Speed information (bits/Ca2+ transient) Wild-type R = 0.167 P = 1.5710–4 7% 9% 24% 60% Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) R = 0.257 P = 3.9510–9 R = –1.6710–4 P = 0.997 2% 3% 2% 3% 29% 7% 9% 24% 14% 60% 66% 81% 8 9 Figure 3. Information about position, speed, and motion direction is independently 0 distributed in the population activity patterns of dDG neurons. Author Contribution Statement 1030 1 2 a, Left panel: Scatter plot showing the distribution of spatial and speed information from 508 3 -0.1 0 0.1 0.2 0.3 0.4 0.5 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% Spatial Info order (from large to small) Speed Info order (from large to small) Direction Info order (from large to small) Reverse order (from small to large) Random order Negative control (using Shuffled Data) Deletion order -0.1 0 0.1 0.2 0.3 0.4 0.5 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining Direction Decoding Decoding accuracy (mean absolute error, radian) Position Decoding Decoding accuracy (mean absolute error, cm) Decoding accuracy (correlation coefficient) Speed Decoding 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% c Wild-type αCaMKII+/– d 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% a Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) Wild-type αCaMKII+/– b Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) R = 0.167 P = 1.5710–4 R = 0.257 P = 3.9510–9 R = –1.6710–4 P = 0.997 R = 0.163 P = 5.4410–3 R = –0.0155 P = 0.793 R = 0.473 P = 1.7310–17 % of Cells Remaining % of Cells Remaining % of Cells Remaining % of Cells Remaining Decoding accuracy (mean absolute error, cm) Decoding accuracy (correlation coefficient) Decoding accuracy (mean absolute error, radian) 2% 3% 2% 3% 1% 2%6% 1% 2% 38% 4% 35% 4% 29% 7% 9% 24% 14% 57% 60% 66% 81% 91% 59% * * * * * * * * a a Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) Speed information (bits/Ca2+ transient) αCaMKII+/– b p (bits/Ca2+ transient) (bits/Ca2+ transient) p (bits/Ca2+ transient) R = 0.163 P = 5.4410–3 R = –0.0155 P = 0.793 R = 0.473 P = 1.7310–17 1% 2%6% 1% 2% 38% 4% 35% 4% 57% 91% 59% Speed information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Direction information (bits/Ca2+ transient) Spatial information (bits/Ca2+ transient) (bits/Ca transient) (bits/Ca transient) R = –0.0155 P = 0.793 R = 0.473 P = 1.7310–17 1% 2%6% 1% 2% 38% 4% 35% % 91% 59% b 108 -0.1 0 0.1 0.2 0.3 0.4 0.5 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% Spatial Info order (from large to small) Speed Info order (from large to small) Direction Info order (from large to small) Reverse order (from small to large) Random order Negative control (using Shuffled Data) Deletion order -0.1 0 0.1 0.2 0.3 0.4 0.5 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining Direction Decoding Decoding accuracy (mean absolute error, radian) Position Decoding Decoding accuracy (mean absolute error, cm) Decoding accuracy (correlation coefficient) Speed Decoding 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% c Wild-type αCaMKII+/– d 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining % of Cells Remaining % of Cells Remaining % of Cells Remaining Decoding accuracy (mean absolute error, cm) Decoding accuracy (correlation coefficient) Decoding accuracy (mean absolute error, radian) * * * * * * *** * Position Decoding Decoding accuracy (mean absolute error, cm) c Wild-type 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% * * c 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% Direction Decoding Decoding accuracy (mean absolute error, radian) 1.3 1.35 1.4 1.45 1.5 1.55 1.6 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining % of Cells Remaining Decoding accuracy (mean absolute error, radian) * Direction Decoding Decoding accuracy mean absolute error, radian) % of Cells Remaining αCaMKII+/– d 13 15 17 19 21 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 1 % of Cells Remaining % of Cells Remaining Decoding accuracy (mean absolute error, cm) * d Decoding accuracy mean absolute error, radian) 9 8 6 5 3 % of Cells Remaining Figure 3. Author Contribution Statement 1030 P = 1.1210–3 () d Position Decoding Wild-type αCaMKII+/– 10 14 18 22 10 14 18 22 d d a -0.3 -0.1 0.1 0.3 0.5 0.7 -0.3 -0.1 0.1 0.3 0.5 0.7 Correlation Coefficient Obs. Shuff. Obs. Shuff. P = 0.619 Speed Decoding e b e Speed (cm/sec) Speed Decoding b Time (sec) Time (sec) Observed Decoded 1.2 1.3 1.4 1.5 1.6 1.7 1.2 1.3 1.4 1.5 1.6 1.7 Mean Absolute Error (radian) Obs. Shuff. P = 0.262 Obs. Shuff. Direction Decoding f Observed Decoded Motion Direction (radian) Direction Decoding c Time (sec) Time (sec) f c Figure 2. Information about position, speed, and motion direction is encoded in the 1083 population activity patterns of dDG neurons. 1084 d, Accuracy of position decoding in 7 wild-type mice (left) and 5 αCaMKII+/– mice (right). The 1096 accuracy of position decoding is reported as the mean absolute error. Black and red lines 1097 represent means, and error bars indicate the standard error of the mean (SEM). 1098 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 1099 e, Accuracy of speed decoding, represented as the correlation coefficient between the observed 1100 and decoded speeds of the mice in each time bin. 1101 1102 f, Accuracy of direction decoding. The decoding error for motion direction is computed as 1103 mean absolute error between the observed and decoded motion directions in each time bin. 1104 1105 1106 1107 . CC-BY 4.0 International license perpetuity. Author Contribution Statement 1030 Information about position, speed, and motion direction is independently 1110 distributed in the population activity patterns of dDG neurons. 1111 a, Left panel: Scatter plot showing the distribution of spatial and speed information from 508 1113 dDG neurons from 7 wild-type mice. Each dot corresponds to an individual neuron. Vertical 1114 and horizontal dashed lines indicate the 95th percentile of spatial and speed information 1115 obtained from shuffled data (same as in Fig. 1d). Pie chart represents the percentage of neurons 1116 a, Left panel: Scatter plot showing the distribution of spatial and speed information from 508 1113 dDG neurons from 7 wild-type mice. Each dot corresponds to an individual neuron. Vertical 1114 and horizontal dashed lines indicate the 95th percentile of spatial and speed information 1115 obtained from shuffled data (same as in Fig. 1d). Pie chart represents the percentage of neurons 1116 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint with spatial and speed information above or below this value. R values are correlation 1117 coefficients. Middle and right panels: Scatter plots showing the distribution of speed and 1118 direction information and of direction and spatial information, respectively. 1119 1120 b, As for panel a, but for 289 dDG neurons from 5 αCaMKII+/– mice. 1121 1122 c, Changes in decoding performance when neurons are individually deleted from the datasets 1123 (left; position decoding, middle; speed decoding, right; direction decoding). Each line shows 1124 the average data from the 7 wild-type mice (P < 0.05, P < 0.01, P < 0.001; Two-way 1125 ANOVA with Tukey-Kramer post hoc tests, vs random order deletion). 1126 1127 d, As for panel c, but showing average data from the 5 αCaMKII+/– mice. 1128 1129 1130 with spatial and speed information above or below this value. R values are correlation 1117 coefficients. Author Contribution Statement 1030 Middle and right panels: Scatter plots showing the distribution of speed and 1118 direction information and of direction and spatial information, respectively. 1119 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint Figure 4 Forced Alternation task a Wild-type αCaMKII+/– Correct choice (%) 4. Free-choice 2. Forced-choice 3. Return 1. Start “Error” 5. Return “Correct” 10 trials/day P = 2.3810–3 b 31 32 Figure 4. Information about current and future LR location is encoded in the population 33 activity patterns of dDG neurons. 34 35 Figure 4 Forced Alternation task a Wild-type αCaMKII+/– Correct choice (%) 4. Free-choice 2. Forced-choice 3. Return 1. Start “Error” 5. Return “Correct” 10 trials/day g LR decoding Accuracy (%) decision period P = 2.3810–3 P = 0.125 P = 1.1910–4 () 30 40 50 60 70 80 90 30 40 50 60 70 80 90 30 40 50 60 70 80 90 30 40 50 60 70 80 90 Obs. Shuff. Obs. Shuff. Obs. Shuff. Obs. Shuff. forced arm period c Wild-Type αCaMKII+/– forced arm period (5sec after forced-turn) decision period (from 6 to 1 sec before decision) Obs. Shuff. LR index Obs. Shuff. e Wild-Type αCaMKII+/– h Cell Count Cell Count LR index LR index ←R-preferential L-preferential→ LR index Wild-Type αCaMKII+/– Wild-Type αCaMKII+/– b LR decoding Accuracy (%) ←R-preferential L-preferential→ ←R-preferential L-preferential→ ←R-preferential L-preferential→ Decision point 12.9 % 13.6 % 15.0 % 13.7 % 16.8 % 18.3 % 9.92 % 7.14 % Cell Count Cell Count Forced R-turn Forced L-turn Cell #14: R-turn preferential Cell #24: not preferential Cell #25: L-turn preferential Future R-choice Future L-choice Cell #3: R-Decision preferential Cell #22: not preferential Cell #43: L-Decision preferential d f Figure Forced Alternation task a 4. Free-choice 2. Forced-choice 3. Return 1. Start “Error” 5. Return “Correct” 10 trials/day Wild-type αCaMKII+/– Correct choice (%) P = 2.3810–3 b a b a Forced Alternation task 4. Free-choice “Error” 5. Return “Correct” 10 trials/day g LR decoding Accuracy (%) P = 0.125 30 40 50 60 70 80 90 30 40 50 60 70 80 90 Obs. Shuff. Obs. Shuff. forced arm period Wild-Type αCaMKII+/– Wild-Type αCaMKII+/– Obs. Shuff. Author Contribution Statement 1030 Cell Count LR index ←R-preferential L-preferential→ LR index ←R-preferential L-preferential→ 12.9 % 18.3 % 9.92 % 7.14 % Cell Count d c forced arm period (5sec after forced-turn) Forced R-turn Forced L-turn Cell #14: R-turn preferential Cell #24: not preferential Cell #25: L-turn preferential g d g c decision period P = 1.1910–4 () 30 40 50 60 70 80 90 30 40 50 60 70 80 90 Obs. Shuff. Obs. decision period (from 6 to 1 sec before decision) Obs. Shuff. LR index e h Cell Count LR index LR index Wild-Type αCaM Wild-Type αCaMKII+/– LR decoding Accuracy (%) ←R-preferential L-preferential→ ←R-preferential L-preferential→ Decision point 13.6 % 15.0 % 13.7 % 16.8 % Cell Count Future R-choice Future L-choice Cell #3: R-Decision preferential Cell #22: not preferential Cell #43: L-Decision preferential f decision period (from 6 to 1 sec before decision) Obs. Shuff. LR index e h Cell Count LR index Wild-Type αCaMKII+/– LR decoding Accuracy (%) ←R-preferential L-preferential→ ←R-preferential L-preferential→ Decision point 13.6 % 15.0 % 13.7 % 16.8 % Cell Count Future R-choice Future L-choice Cell #3: R-Decision preferential Cell #22: not preferential Cell #43: L-Decision preferential f decision period (from 6 to 1 sec before decision) e Decision point Future R-choice Future L-choice Cell #3: R-Decision preferential Cell #22: not preferential Cell #43: L-Decision preferential Obs. Shuff. LR index Cell Count LR index Wild-Type αCaMKII+/– ←R-preferential L-preferential→ ←R-preferential L-preferential→ 13.6 % 15.0 % 13.7 % 16.8 % Cell Count f decision period P = 1.1910–4 () 30 40 50 60 70 80 90 30 40 50 60 70 80 90 Obs. Shuff. Obs. Shuff. h Wild-Type αCaMKII+/– LR decoding Accuracy (%) f h h e 1131 1132 Figure 4. Information about current and future LR location is encoded in the population 1133 activity patterns of dDG neurons. 1134 1135 a, Forced-alternation task in a modified T-maze test. Each trial consists of a forced-choice run 1136 followed by a free-choice run. In the forced-choice run, the mouse is forced to turn one of two 1137 randomly selected directions (1. Start→2. Forced choice), whereas in the free-choice run, the 1138 mouse can choose to turn left or right (3. Return→4. Free choice). This task exploits the 1139 behavioural tendency of mice to choose the arm opposite the one previously chosen. Author Contribution Statement 1030 The centre lines, box boundaries and whiskers for the boxplot indicate 1146 median, upper and lower quartile, and maximum and minimum of the data. 1147 1148 c, The forced arm period was defined as the 5 sec period after the mouse performed the forced 1149 turn (left). Color-coded cell activity maps of three representative dDG neurons from wild-type 1150 mice (right). These maps show the density of mouse locations where Ca2+ transients were 1151 detected in the cases of L-turn or R-turn in all 50 trials. Each map is normalized to each 1152 neuron’s maximum Ca2+ transient rate and smoothed with a Gaussian kernel (σ = 2.0 cm). 1153 1154 d, Distributions of LR indices calculated from the recordings of the observed neurons (red 1155 curve, “Obs.”) and from shuffled data (grey curve, “Shuff.”; 1000 shuffles per cell; counts 1156 normalized by number of shuffles) in the arm period, which corresponds to the 5 sec following 1157 the forced turn. All 294 neurons from 7 wild-type mice and 131 neurons from 4 αCaMKII+/– 1158 mice are shown in histograms. The dashed line shows the 95th percentile of the distribution of 1159 the shuffled data, and the numbers on the lines represent the percentage of observed neurons 1160 exceeding this value. 1161 1162 e, The decision period was defined as the 6 to 1 sec period prior to the free-choice turn (left). 1163 As for panel (c), but for color-coded cell activity maps of representative dDG neurons in the 1164 decision period (right). 1165 1166 f, As for panel (d), but for LR indices in decision period, which corresponds to the 6 to 1 sec 1167 before the turn decision is made. 1168 1169 g, LR decoding accuracy in wild-type mice and αCaMKII+/– mice from the population activity 1170 patterns of dDG neurons in the arm period. The grey (left panel, wild-type mice) and pale red 1171 lines (right panel, αCaMKII+/– mice) show the decoding accuracy of the individual mice in 1172 each group. Black (left panel, wild-type mice) and red (right panel, αCaMKII+/– mice) lines 1173 indicate the average decoding accuracy of all mice in each group. Error bars indicate SEM. 1174 1175 h As for panel (e) but for the decision period 1176 presented to it in the forced choice run, the trial is considered correct ; otherwise, it is 1142 considered an “error”. Author Contribution Statement 1030 Therefore, 1140 if the direction that the mouse chooses in the free-choice run is opposite to the direction 1141 LR index ←R-preferential L-preferential→ 1131 1132 Figure 4. Information about current and future LR location is encoded in the population 1133 activity patterns of dDG neurons. 1134 1135 LR index 1131 1132 Figure 4. Information about current and future LR location is encoded in the population 1133 activity patterns of dDG neurons. 1134 1135 a, Forced-alternation task in a modified T-maze test. Each trial consists of a forced-choice run 1136 followed by a free-choice run. In the forced-choice run, the mouse is forced to turn one of two 1137 randomly selected directions (1. Start→2. Forced choice), whereas in the free-choice run, the 1138 mouse can choose to turn left or right (3. Return→4. Free choice). This task exploits the 1139 behavioural tendency of mice to choose the arm opposite the one previously chosen. Therefore, 1140 if the direction that the mouse chooses in the free-choice run is opposite to the direction 1141 LR index Figure 4. Information about current and future LR location is encoded in the population 1133 activity patterns of dDG neurons. 1134 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint presented to it in the forced-choice run, the trial is considered “correct”; otherwise, it is 1142 considered an “error”. 1143 1144 b, The correct choice rate (%) in the forced-alternation task in 7 wild-type mice and 4 1145 αCaMKII+/– mice. h, As for panel (e), but for the decision period. 6 Author Contribution Statement 1030 CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 1177 a c Wild-type Wild-type R = 0.2281 P = 7.9110–5 LR index (decision period) ←R-preferential L-preferential→ 6% 14% 23% 57% 1178 1179 Figure 5. Information about current and future LR locations is widely distributed in the 1180 population activity patterns of dDG neurons. Author Contribution Statement 1030 1181 a b LR index (decision period) ←R-preferential L-preferential→ % of Cells Remaining Decoding accuracy (%) LR decoding (forced arm period) LR decoding (decision period) 40 50 60 70 80 90 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 90 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% LR index (forced arm period) ←R-preferential L-preferential→ c d e LR decoding (forced arm period) LR decoding (decision period) Decoding accuracy (%) % of Cells Remaining Wild-type αCaMKII+/– Wild-type αCaMKII+/– Decreasing LR index order (forced arm period, in absolute value) Decreasing LR index order (decision period, in absolute value) Increasing LR index order (in absolute value) Random order Negative control (using Shuffled Data) Deletion order R = 0.2281 P = 7.9110–5 R = – 0.1210 P = 0.1687 LR index (decision period) ←R-preferential L-preferential→ LR index (forced arm period) ←R-preferential L-preferential→ 6% 14% 23% 14% 14% 16% -30 -20 -10 0 10 20 30 40 0.65 0.75 0.85 0.95 LR Decoding Accuracy (decision period) (Observed–Shuffled) (%) Pattern similarity (Population Vector Overlap (PVO) between forced arm and decision period) R = –0.8738 WT (n = 7mice) αCaMKII+/– (n = 4mice) 57% 56% * * * * * * a Decoding accuracy (%) LR decoding (forced arm period) LR decoding (decision period) 40 50 60 70 80 90 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% % of Cells Remaining Wild type * * * * Wild-type % of Cells Remaining % of Cells Remaining b LR index (decision period) ←R-preferential L-preferential→ LR index (forced arm period) R f ti l L f ti l d αCaMKII+/– αCaMKII+/– R = – 0.1210 P = 0.1687 LR index (forced arm period) ←R-preferential L-preferential→ 14% 14% 16% 56% % of Cells Remaining 40 50 60 70 80 90 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% LR decoding (forced arm period) LR decoding (decision period) Decoding accuracy (%) % of Cells Remaining Decreasing LR index order (forced arm period, in absolute value) Decreasing LR index order (decision period, in absolute value) Increasing LR index order (in absolute value) Random order Negative control (using Shuffled Data) Deletion order * * 40 50 60 70 80 90 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 40 50 60 70 80 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% d LR decoding (forced arm period) LR decoding (decision period) Decoding accuracy (%) g αCaMKII+/– 4% % * *** d d b αCaMKII+/– % of Cells Remaining 1178 1179 e Decreasing LR index order (forced arm period, in absolute value) Decreasing LR index order (decision period, in absolute value) Increasing LR index order (in absolute value) Random order Negative control (using Shuffled Data) -30 -20 -10 0 10 20 30 40 0.65 0.75 0.85 0.95 LR Decoding Accuracy (decision period) (Observed–Shuffled) (%) Pattern similarity (Population Vector Overlap (PVO) between forced arm and decision period) R = –0.8738 WT (n = 7mice) αCaMKII+/– (n = 4mice) e WT (n = 7mice) αCaMKII+/– (n = 4mice) Figure 5. Author Contribution Statement 1030 1143 1144 b, The correct choice rate (%) in the forced-alternation task in 7 wild-type mice and 4 1145 αCaMKII+/– mice. The centre lines, box boundaries and whiskers for the boxplot indicate 1146 median, upper and lower quartile, and maximum and minimum of the data. 1147 1148 c, The forced arm period was defined as the 5 sec period after the mouse performed the forced 1149 turn (left). Color-coded cell activity maps of three representative dDG neurons from wild-type 1150 mice (right). These maps show the density of mouse locations where Ca2+ transients were 1151 detected in the cases of L-turn or R-turn in all 50 trials. Each map is normalized to each 1152 neuron’s maximum Ca2+ transient rate and smoothed with a Gaussian kernel (σ = 2.0 cm). 1153 1154 d, Distributions of LR indices calculated from the recordings of the observed neurons (red 1155 curve, “Obs.”) and from shuffled data (grey curve, “Shuff.”; 1000 shuffles per cell; counts 1156 normalized by number of shuffles) in the arm period, which corresponds to the 5 sec following 1157 the forced turn. All 294 neurons from 7 wild-type mice and 131 neurons from 4 αCaMKII+/– 1158 mice are shown in histograms. The dashed line shows the 95th percentile of the distribution of 1159 the shuffled data, and the numbers on the lines represent the percentage of observed neurons 1160 exceeding this value. 1161 1162 e, The decision period was defined as the 6 to 1 sec period prior to the free-choice turn (left). 1163 As for panel (c), but for color-coded cell activity maps of representative dDG neurons in the 1164 decision period (right). 1165 1166 f, As for panel (d), but for LR indices in decision period, which corresponds to the 6 to 1 sec 1167 before the turn decision is made. 1168 1169 g, LR decoding accuracy in wild-type mice and αCaMKII+/– mice from the population activity 1170 patterns of dDG neurons in the arm period. The grey (left panel, wild-type mice) and pale red 1171 lines (right panel, αCaMKII+/– mice) show the decoding accuracy of the individual mice in 1172 each group. Black (left panel, wild-type mice) and red (right panel, αCaMKII+/– mice) lines 1173 indicate the average decoding accuracy of all mice in each group. Error bars indicate SEM. 1174 1175 h, As for panel (e), but for the decision period. 1176 . Author Contribution Statement 1030 Information about current and future LR locations is widely distributed in the 1180 population activity patterns of dDG neurons. 1181 . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint . CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted November 15, 2020. ; https://doi.org/10.1101/2020.06.09.141572 doi: bioRxiv preprint 1191 c, d, Changes in LR decoding performance (left panels, arm period; right panels, decision 1192 period) when neurons were individually deleted from the datasets for wild-type mice (c) and 1193 αCaMKII+/– mice (d). The horizontal axis represents the percentage of remaining neurons that 1194 are used for population decoding after neurons are deleted. The vertical axis shows the LR 1195 decoding accuracy (%) in the arm period (left panels) and the decision period (right panels). 1196 Each line shows the average of 7 wild-type mice (c) and 4 αCaMKII+/– mice (d) (P < 0.05, 1197 P < 0.01, *P < 0.001; Two-way ANOVA with Tukey-Kramer post hoc tests, vs random 1198 order deletion). 1199 1200 e, Scatter plot showing the correlation between the PVO (forced arm period × decision period) 1201 and LR decoding accuracy in the decision period, which is computed as the decoding accuracy 1202 obtained from observed data minus those of shuffled data (Obs–Shuffled, %). The individual 1203 black and red dots correspond to individual wild-type and αCaMKII+/– mice, respectively. 1204 1205 (5 figures) 1206 1191 c, d, Changes in LR decoding performance (left panels, arm period; right panels, decision 1192 period) when neurons were individually deleted from the datasets for wild-type mice (c) and 1193 αCaMKII+/– mice (d). The horizontal axis represents the percentage of remaining neurons that 1194 are used for population decoding after neurons are deleted. The vertical axis shows the LR 1195 decoding accuracy (%) in the arm period (left panels) and the decision period (right panels). Author Contribution Statement 1030 1196 Each line shows the average of 7 wild-type mice (c) and 4 αCaMKII+/– mice (d) (P < 0.05, 1197 P < 0.01, P < 0.001; Two-way ANOVA with Tukey-Kramer post hoc tests, vs random 1198 order deletion). 1199 1200 e, Scatter plot showing the correlation between the PVO (forced arm period × decision period) 1201 and LR decoding accuracy in the decision period, which is computed as the decoding accuracy 1202 obtained from observed data minus those of shuffled data (Obs–Shuffled, %). The individual 1203 black and red dots correspond to individual wild-type and αCaMKII+/– mice, respectively. 1204 1205 (5 figures) 1206 1191 c, d, Changes in LR decoding performance (left panels, arm period; right panels, decision 1192 period) when neurons were individually deleted from the datasets for wild-type mice (c) and 1193 αCaMKII+/– mice (d). The horizontal axis represents the percentage of remaining neurons that 1194 are used for population decoding after neurons are deleted. The vertical axis shows the LR 1195 decoding accuracy (%) in the arm period (left panels) and the decision period (right panels). 1196 Each line shows the average of 7 wild-type mice (c) and 4 αCaMKII+/– mice (d) (P < 0.05, 1197 P < 0.01, *P < 0.001; Two-way ANOVA with Tukey-Kramer post hoc tests, vs random 1198 order deletion). 1199 1200 e, Scatter plot showing the correlation between the PVO (forced arm period × decision period) 1201 and LR decoding accuracy in the decision period, which is computed as the decoding accuracy 1202 obtained from observed data minus those of shuffled data (Obs–Shuffled, %). The individual 1203 black and red dots correspond to individual wild-type and αCaMKII+/– mice, respectively. 1204 1205 (5 figures) 1206	6,276
https://openalex.org/W4282941888	OpenAlex	Open Science	CC-By	2,022	Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes	Mohammad ALOBAIDY	English	Spoken	8,158	15,650	Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes Mohammad Gazy ALOBAIDY1, Zübeyir AĞIRAĞAÇ2, Şeyda ZORER ÇELEBİ3 1Van Yuzuncu Yil University, Institute of Natural and Applied Science PhD, 65000, Van, Turkey 2,3Van Yuzuncu Yil University, Agriculture Faculty, Field Crops Department, 65080, Van, Turkey Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes Mohammad Gazy ALOBAIDY1, Zübeyir AĞIRAĞAÇ2, Şeyda ZORER ÇELEBİ3 1Van Yuzuncu Yil University, Institute of Natural and Applied Science PhD, 65000, Van, Turkey 2,3Van Yuzuncu Yil University, Agriculture Faculty, Field Crops Department, 65080, Van, Turkey Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes Mohammad Gazy ALOBAIDY1, Zübeyir AĞIRAĞAÇ2, Şeyda ZORER ÇELEBİ3 1Van Yuzuncu Yil University, Institute of Natural and Applied Science PhD, 65000, Van, Turkey 2,3Van Yuzuncu Yil University, Agriculture Faculty, Field Crops Department, 65080, Van, Turkey 1htt / id /0000 0002 9894 9673 2htt / id /0000 0003 1414 1472 3htt / id /0000 0003 1278 1994 orcid.org/0000-0002-9894-9673, 2https:/orcid.org/0000-0003-1414-1472, 3https:/orcid.org/0000-0003-1278-1994 Corresponding author e-mail: seydazorer@yyu.edu.tr Abstract: In agriculture, the use of environmentally friendly materials both in place of and alongside chemical ones is continuously increasing. Two field experiments were conducted during the 2019 and 2020 seasons to investigate the effects of foliar application of three doses (5.0, 7.5, and 10 g L-1) of active dry yeast were sprayed on maize (Zea mays L. Cv. Tuano) at two intervals, the first 54 days after planting (DAP), and the second 15 days later. Plants were grown for silage purposes under alkaline soil conditions at the Experimental and Research Station of the Field Crops Dep., Van Yuzuncu Yıl University-(VYYU), Turkey. Measurements were taken three times at the vegetative stage 64, 74, and 84-DAP, and one times at the dough stage 117-123 DAP. Morphologically, the results showed that the foliar application of different doses of yeast increased the plant height (cm), stem, leaves, and cobs weight per plant and total plant weight (g), number of cobs plant-1 (piece), and green and dry herbage yield (ton da-1) at 117- 123 DAP during the two seasons. In contrast, there was no significant increase in plant height (cm), chlorophyll as SPAD (The Soil Plant Analysis Development) value, and the number of leaves per plant at 64 DAP. Chemically, the spraying of the yeast improved the P, K, Ca, Mg, Fe, Cu, Se, and Zn concentrations at 117- 123 DAP as well as chlorophyll content at 74 and 84 DAP compared to the control. Keywords Alkaline soil, Active dry yeast, Maize/Corn plant, Silage To Cite: Alobaidy, M G, Ağırağaç, Z, Zorer Çelebi, Ş, 2022. Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes. Yuzuncu Yil University Journal of Agricultural Sciences, 32(2): 310-319. DOI: https://doi.org/10.29133/yyutbd.1048847 Article Info Article Info Received: 27.12.2021 Accepted: 06.05.2022 Online published: 15.06.2022 DOI: 10.29133/yyutbd.1048847 Received: 27.12.2021 Accepted: 06.05.2022 Online published: 15.06.2022 DOI: 10.29133/yyutbd.1048847 Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes Mohammad Gazy ALOBAIDY1, Zübeyir AĞIRAĞAÇ2, Şeyda ZORER ÇELEBİ3 1Van Yuzuncu Yil University, Institute of Natural and Applied Science PhD, 65000, Van, Turkey 2,3Van Yuzuncu Yil University, Agriculture Faculty, Field Crops Department, 65080, Van, Turkey From this study, it could be concluded that the highest values of the studied parameters were recorded when active dry yeast was used at a dose of 10 g L-1 on maize under high pH soil. Yuzuncu Yil University Journal of Agricultural Sciences, Volume: 32, Issue: 2, 30.06.2022 Yuzuncu Yil University Journal of Agricultural Sciences, Volume: 32, Issue: 2, 30.06.2022 Yuzuncu Yil University Journal of Agricultural Sciences (Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi) https://dergipark.org.tr/en/pub/yyutbd ISSN: 1308-7576 e-ISSN: 1308-7584 Research Article e-ISSN: 1308-7584 Yuzuncu Yil University Journal of Agricultural Sciences (Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi) https://dergipark.org.tr/en/pub/yyutbd 1. Introduction Maize or corn is one of the most important cereal crops in the world. It is grown under a wide range of agronomic and environmental circumstances (Khan et al., 2014; Rouf Shah et al., 2016). It is the main source of energy and food source for humans and animals in addition to its content of natural phytochemical compounds such as pigments, substantial fats, and starch (Rouf Shah et al., 2016; Kaul et al., 2019). Moreover, over the past few decades, the silage of corn has become the main fodder ingredient in the dairy cows' rations (Khan et al., 2014). The cultivation area of corn produced for silage in Turkey is approximately 4.7 million decares. Total silage corn production is 23.2 million tons, and the silage yield is 4915 kg da-1 on average, although it may vary by region (Acar et al., 2020). 73% of 310 YYU J AGR SCI 32 (2): 310-319 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes YYU J AGR SCI 32 (2): 310-319 Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes ( ) aidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes corn production in the world and 90% in developed countries is used for animal feeding. In Turkey, 70% of corn production, which ranks third after wheat and barley, is used for animal nutrition (Öz et al., 2017). corn production in the world and 90% in developed countries is used for animal feeding. In Turkey, 70% of corn production, which ranks third after wheat and barley, is used for animal nutrition (Öz et al., 2017). Nutrient materials solubility and availability, in addition to soil microorganism activity, mainly depend on soil reaction (pH). Most micronutrients, for instance, are less available in neutral-alkaline soils compared to acid soils, which are considered the preferred conditions to plant growth (Lončarić et al., 2008). In general, under alkaline soil conditions, the availability of most macronutrients is increased. In contrast, the availability of micronutrients and some macronutrients such as phosphorus is decreased. This decrease leads to a negative impact on plant growth and yield (Jiang et al., 2017). 1. Introduction This research aimed to study the effect of spraying bio-fertilizers (active dry bread yeast) in improving the growth of maize crops for silage purposes under alkaline soil conditions. 1. Introduction According to the salinity and alkalinity criteria used in the Turkey Improved Soil Map studies, there is a problem with salinity and alkalinity in an area of 1.518.722 ha in Turkey. Where 0.5% is alkaline, 8% is slightly salty alkali, and 17.5% is salty alkali. The barren lands are equivalent to 5.48% of the total cultivated agricultural lands (Sönmez and Beyazgül, 2008). g ( y g , ) The use of bio/organic fertilizers is increasing all over the world for environmental and human health safeguard purposes, which has become endangered as a result of the excessive use of chemical fertilizers and pesticides (Agamy et al., 2013). Bio-stimulants influence plant metabolism if it used in low concentrations through prompting of natural plant growth regulators driving and vigor, reinforcement of nutrients absorption, enhancement of root growth, and increasing resistance to stress as abnormal conditions. In addition, these materials could be used as systemic agents when they are applied as a foliar application (Hassan et al., 2020). Abbas (2013) and Alsaady et al. (2020) concluded that the application of bio-stimulants such as yeast in small quantities affected several metabolic processes and promoted plant growth, productivity, and development through increased photosynthesis, natural hormones, ion uptake, and protein synthesis as well as a relative increase in the availability of the micronutrients in the soil. It may also increase antioxidants and enhance metabolism and water holding capacity. Yeast Saccharomyces cerevisiae extract, which is plant nutritious as well as an, environmentally friendly, and convenient to use, has a feature over common plant growth regulators and soil conditioners (Xi et al., 2019) and includes natural plant hormones i.e., free and associated IAA, free GA3 and associated GA3 (Twfiq, 2010) and vitamins, amino acids, necessary macro and micronutrients for plant growth, and growth stimuli such as Cytokinins (Medani and Taha, 2015) and it is counted as a natural source of biostimulants that motivate some enzymes, cell expansion and division, synthesis of pigments, and nucleic acid and protein formation (Wanas, 2006), and also it releases CO2 which leads to improvement of photosynthesis (Kurtzman and Fell, 2005). Al-Shaheen et al. (2019) stated that the application of active dry yeast had positive effects on corn plants. All sprayed maize plants with active dry yeast extract produced the highest yield rate of all the studied characters. 2. Material and Methods Two field experiments were performed to evaluate the effects of active dry yeast on the growth, morphological changes, and chemical constituents of maize (Zea mays L. Cv. Tuano) for silage purposes. The experiments were carried out in the experimental field of VYYU-Campus, Fac. Agric., Field Crops Department as well as in the Science Application and Research Center labs, Van, Turkey, during the two successive summer seasons, 2019 and 2020. Some climatic data of the province of Van during the years of the experiment are given in Table 1. The average temperature in Van city in the 2019 and 2020 seasons were higher than the average temperature for the long years. In contrast, total precipitations in the experimental seasons (2019 and 2020) were lower than the long-term ones. The same trend was recorded for the average humidity in Van city during the 2019 and 2020 seasons were lower compared to long years average, as shown in Table 1. 311 YYU J AGR SCI 32 (2): 310-319 ( ) Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Pur Table 1. Some climate values of the experimental area for the years 2019 and 2020 and the long-term average Average Temperature (oC) Total Precipitation (mm) Average Humidity (%) Month 2019 2020 Long Years 2019 2020 Long Years 2019 2020 Long Years April 7.2 8.6 8.4 36.2 50.9 57.4 66.1 65.4 59.3 May 15.4 14.5 13.4 15.3 27.8 45.3 51.9 54.0 55.1 June 21.4 19.3 18.8 7.2 13.4 16.4 45.4 44.4 47.1 July 23.0 23.0 22.7 0.4 17.9 6.9 39.0 46.4 42.3 August 23.7 21.6 22.9 0.9 10.0 5.3 40.2 44.5 40.5 September 18.8 20.1 18.3 0.8 5.6 20.4 43.9 41.3 43.9 October 13.4 13.3 12.0 24.1 1.8 48.2 52.9 47.2 57.3 Mean/Total 17.6 17.2 16.6 85 127 199 48.4 49.0 49.3 * Van Meteorology Regional Directorate records. Some climate values of the experimental area for the years 2019 and 2020 and the long-term average* * Van Meteorology Regional Directorate records. * Van Meteorology Regional Directorate records. Soil samples were randomly taken each year before cultivation at the two depths of 0-20 and 20-40 cm, and were subjected to some physical and chemical analysis (VYYU, Fac. Agric., Dep. Soil Science and Plant Nutrition) were illustrated in Tables 2. 2.1. Data Recorded In both experiments, three samples on five plants per plot, i.e., at 64, 74, and 84-DAP, were read from each treatment 10 days after each application. In which each one was represented by three replicates for the morphological studies (plant height (cm), number of leaves per plant), and total chlorophyll SPAD. At the dough stage (117-123 DAP), a sample was taken to record the following: plant height (cm), number of leaves per plant, number of cobs per plant (piece), stem, leaves, cobs weight per plant (g), plant weight (g), stem diameter (mm), and green and dry herbage yield (ton da-1). Macronutrients (P, K, Ca, Mg) and micronutrients (Fe, Cu, Mn, Mo, Se, Zn) concentrations were determined using by wet digestion inductively coupled plasma-optical emission spectrometer (ICP- OES) and atomic absorption spectrophotometer (AAS) in VYYU, Science Application and Research Center labs according to Association of official analytical chemists (1995, chap. 50). 2. Material and Methods Experiments soil pH ranged between 7.94 and 8.16 in the 0-20 cm and 20-40 cm, respectively. The experiment's soil was sandy-clay in VYYU- Campus, as shown in Table 2. Table 2. Some physical and chemical analysis of the soil experimental field Deep pH Sat. Clay (%) Silt (%) Sand (%) Lime (%) Ca+Mg (me 100g-1) CEC (me 100g-1) Organic Matter (%) 0-20 7.94 35.08 20.95 43.97 21.06 13.30 16.00 1.87 20-40 8.16 31.76 18.88 49.39 21.41 14.25 17.00 1.69 Table 2. Some physical and chemical analysis of the soil experimental field ize seeds were obtained from the Van Directorate of Provincial Agriculture and Forestry. h Maize seeds were obtained from the Van Directorate of Provincial Agriculture and Forestry. Seeds were sown on 15th May in both seasons, and all experimental units were 5.0×3.5 m, containing 6 rows (70 cm between rows) and 12 cm between plants. The distance between plots has been arranged as 2 m and between blocks as 3 m. With the planting, fertilizer will be applied to all plots at the rate of 8 kg N da-1 at the first batch and 8 kg P2O5 da-1. When corn is 50-60 cm tall, second dose of nitrogen fertilizer will be applied at the rate 10 kg N da-1 (Sabancı, 2015). Hoeing has been done two times to cover the bottom of the plant stem, aerate the soil, and fight weeds. Plants were irrigated with a sprinkler irrigation system. The active dry yeast was prepared from bread yeast (Saccharomyces cerevisiae), by solving dry yeast in hot water, then added sugar in a ratio of 1:1 and kept in a warm place for 24 hours, according to Morsi et al. (2008). Three different doses of active dry bread yeast (5.0, 7.5, and 10.0 g L- 1), as well as control plots treated with tab water, were sprayed on maize plants under alkali soil stress two times started at 54 DAP and repeated after 15 days. 3.1. Vegetative Characters The effects of active dry yeast applications on plant height (cm), number of leaves per plant, and leaf chlorophyll content (SPAD value) are given in Table 3. Progressive significant height of maize plant, number of leaves per plant, and SPAD measurements were achieved with the foliar application of active dry yeast. When 10 g of yeast was sprayed on the plant above soil, the plant height and number of leaves of maize reached the highest records of the second and third vegetative stages than the yeast- free treatment, control, while the differences between effects of yeast concentrations at 64 DAP were insignificant. Table 3. Effect of dry yeast foliar application on vegetative characters of maize grown under high pH soil conditions at 64, 74, and 84 DAP during 2019 and 2020 seasons Treatments Plant height (cm) Number of leaves per plant SPAD Value Spray 64 Days After Planting Control 137.83±26.84 NS 10.20±0.872 NS 41.07±5.06 NS M1: 5.0 g L-1 137.03±9.99 10.30±0.755 38.82±2.62 M2: 7.5 g L-1 146.50±19.02 10.80±0.361 43.14±3.80 M3: 10.0 g L-1 131.97±17.60 10.90±0.265 42.28±2.13 Mean 138.33 10.55 41.33 P-Value 0.4758 0.2774 0.5168 S.E.D.* 9.500 0.505 2.930 74 Days After Planting Control 163.15±10.79 c 11.84±0.703 NS 36.11±3.47 c M1: 5.0 g L-1 196.36±12.48 b 12.03±0.701 42.19±0.68 b M2: 7.5 g L-1 204.23±17.62 ab 12.33±0.351 43.87±1.99 b M3: 10.0 g L-1 216.47±13.83 a 12.73±0.252 48.45±1.33 a Mean 195.05 12.23 42.66 P-Value 0.0003 0.0967 0.0001 S.E.D. 11.360 0.442 1.740 84 Days After Planting Control 196.31±11.18 d 13.33±0.635 NS 38.55±1.53 c M1: 5.0 g L-1 220.76±11.14 c 13.43±0.751 45.03±1.14 b M2: 7.5 g L-1 233.13±11.33 b 13.55±0.253 47.25±0.49 b M3: 10.0 g L-1 247.67±14.02 a 14.29±0.278 51.08±0.70 a Mean 224.47 13.65 45.48 P-Value 0.0001 0.0781 0.0001 S.E.D. 9.780 0.430 0.855 The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. S.E.D.: Standard errors of differences of means, NS: insignificant difference. Table 3. Effect of dry yeast foliar application on vegetative characters of maize grown under soil conditions at 64, 74, and 84 DAP during 2019 and 2020 seasons The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means, NS: insignificant difference. 2.2. Experimental design and statistical analysis The field experiments were arranged in the Randomized Complete Block Design (RCBD) with three replications. Three different doses of active dry bread yeast (Saccharomyces cerevisiae), as well 312 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes YYU J AGR SCI 32 (2): 310-319 YYU J AGR SCI 32 (2): 310-319 id t l / D t i ti Eff t f A ti D Y t M h l i l d Ch i l C t f M i Pl t G i Alk li S il f Sil P YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Pur as control, were sprayed on maize plants. All data recorded were subjected to normal statistical analysis (One-Sample Kolmogorov-Smirnov Test), and the test distribution is normal. One-way analysis of variance (ANOVA) and Duncan multiple comparison test were used to evaluate the treatment effects. Then the combined analysis of the two seasons was done according to Steel and Torrie (1960). The differences among treatment mean values were compared using Duncan multiple range test, and a p- value of 5% was used for statistical significance as reported by Gomez and Gomez (1984). All statistical analysis was performed by using the analysis of variance (ANOVA) technique of SAS Software package Stat-view-9.0.1 (SAS Institute, 2002). 3.2. Morphological Characters at the Dough Stage The impact of spraying active dry yeast on morphological characters of maize at the dough stage for the mean of the two seasons (2019 and 2020) is indicated in Tables 4 and 5. Using the 5.0, 7.5, and 10.0 g L-1doses of dry yeast have a positive effect on the maize plant growth for silage purposes in comparison with the control treatment (without yeast spraying). Plant height, stem weight per plant, and plant leaf weight in the dough stage differed significantly according to doses of yeast and control applications (p ≤ 0.05). The greatest value was detected in corn treated with active dry yeast (10 g L-1), followed by (7.5 g L-1), then by (5 g L-1), and the smallest values were in control non-sprayed plants. While no significant difference was detected between the M2: 7.5 g L-1and M3: 10.0 g L-1of dry yeast on the plant leaf weight (Table 4). Table 4. Effect of dry yeast foliar application on morphological characters of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons Treatments Plant height (cm) Number of leaves per plant Stem diameter (mm) Stem weight plant-1 (g) Leaves weight plant-1 (g) Spray Control 220.40±26.84 d 13.93±0.25 NS 24.56±1.27 NS 220.53±53.1 d 103.03±14.99 c M1: 5.0 g L-1 246.97±9.99 c 14.24±0.90 23.22±0.99 289.87±58.3 c 129.53±12.45 b M2: 7.5 g L-1 266.03±19.02 b 14.68±0.11 22.65±3.38 316.73±58.9 b 141.60±6.77 ab M3: 10.0 g L-1 282.91±17.6 a 14.68±0.01 24.75±1.19 396.73±73.2 a 158.00±3.50 a Mean 254.08 14.38 23.79 305.97 133.04 P-Value 0.0001 0.1103 0.5259 0.0001 0.0009 S.E.D.** 15.770 0.387 1.592 50.100 8.540 The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. S.E.D.: Standard errors of differences of means, NS: insignificant difference. Effect of dry yeast foliar application on morphological characters of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means, NS: insignificant difference. 3.1. Vegetative Characters The illustrated parameters (Table 3) are in agreement with the findings presented by (Abu Khouder et al., 2019; Al-Shaheen et al., 2019; Al-madhagi., 2019); they pointed out that the application of yeast increased the plant height, number of leaves per plant and rate of chlorophyll in the plant leaves. In this context, the highly impactful CO2 release was due to the use of active dry yeast, which subsequently leads to an increase the net plant photosynthesis (Ferguson et al., 1995; Vas and Papanas, 313 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes YYU J AGR SCI 32 (2): 310-319 YYU J AGR SCI 32 (2): 310-319 id l / D i i Eff f A i D Y M h l i l d Ch i l C f M i Pl G i Alk li S il f Sil P YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Pur 2019). The increase of the concentration of chlorophyll when the foliar application of yeast is because it contains the natural hormones that motivate the chlorophyll formation. Yeast includes Cytokinin, which is vital for chloroplast up growth and is the backbone of chlorophyll synthesis EzzEL-Din and Hendawy (2010). Moreover, activating of plant vegetative growth using dry yeast could be due to its effect on the transmission of the nutritional signal that constitutes growth regulators (El-Ghadban et al., 2003). 3.2. Morphological Characters at the Dough Stage Green and dry herbage yield (ton da-1) characteristics recorded significant responses when maize plants were treated by the different levels of active dry yeast in comparison to not sprayed plants in the average of the two seasons, 2019 and 2020, 314 YYU J AGR SCI 32 (2): 310-319 Al b id t l / D t i ti Eff t f A ti D Y t M h l i l d Ch i l C t YYU J AGR SCI 32 (2): 310-319 id l / D i i Eff f A i D Y M h l i l d Ch i l C f M i Pl G i Alk li S il f Sil P YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silag as shown in Table 5. Although there is a significantly green herbage yield (ton da-1) difference between control and plants sprayed by yeast, there are slight differences (not significant) between the concentrations of active dry yeast themselves. For dry herbage yield (ton da-1), generally, the differences between the concentrations of active dry yeast themselves and between yeast doses and control were significant in the dough stage during the 2019 and 2020 seasons. In this context, there are many studies on the application of yeast in different plants such as maize, sunflower, and broad bean. There are results that plant growth is positively affected the growth characteristics as well as yield and its components reported by some researchers (Seadh et al., 2015; Altunlu et al., 2019; Al-Shaheen et al., 2019; Abed and Zeboon, 2020; Al-Ani et al., 2020). The results mentioned in Table 4 are in the same trend. The increase may be due to the effect of the yeast spray because it can produce phytohormones such as Auxins, Gibberellic acid, Cytokinins, and some vitamins (EL-Kholy et al., 2007), which by their physiological action are stimulated the elongation and cell division. 3.2. Morphological Characters at the Dough Stage Besides, they contain amino acids and some mineral elements such as nitrogen and others that enhance plant growth, and this result is consistent with what (Ahmed et al., 2011). In this context, the yeast catalyzes the release of CO2 from fermentation, which will eventually contribute to photosynthesis and increase the products resulting from the process (Khalil and Ismael, 2010). Table 5. Effect of dry yeast foliar application on morphological characters of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons Treatments Number of cobs plant-1 (g) Cobs weight plant-1 (g) Plant weight (g) Green herbage yield (ton da-1) Dry herbage yield (ton da-1) Spray Control 1.07±0.116 b 140.20±16.56 d 463.77±80.5 d 7.50±0.98 b 1.63±0.18 d M1: 5.0 g/L 1.23±0.322 b 280.40±40.6 c 699.80±101.9 c 8.79±0.57 a 2.23±0.14 c M2: 7.5 g/L 1.30±0.100 ab 314.40±24.3 b 772.73±85.5 b 8.90±0.28 a 2.63±0.32 b M3: 10.0 g/L 1.57±0.058 a 385.30±44.4 a 940.03±87.2 a 9.30±0.33 a 3.18±0.35 a Mean 1.29 280.07 719.08 8.62 2.41 P-Value 0.0175 0.0001 0.0001 0.0001 0.0017 S.E.D.** 0.1472 27.320 72.800 0.4945 0.2135 The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. S.E.D.: Standard errors of differences of means. Effect of dry yeast foliar application on morphological characters of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means. 3.2. Morphological Characters at the Dough Stage On the other hand, there was not a significant difference between maize plants treated with all dry yeast concentrations and control on the number of leaves per plant and stem diameter (mm) of maize plants grown under alkaline soil conditions at the dough stage during 2019 and 2020 seasons as Table 4 shown. This may explain that these traits are associated with the genetic cultivar of corn. It is well known that the actual performance of any cultivar depends on its genetic parameters interacting with all surrounded environmental conditions (Saleh et al., 2018). For the cobs number per plant, the data presented in Table 5 indicated that because of the applying active dry yeast to maize, the highest value of number of cobs per plant (1.57) was recorded at 10 g L-1, while the lowest value of number of cobs per plant (1.07) was obtained as a result of the non-application of dry yeast 0 g L-1 was obtained in the control plots. On the other hand, it is seen that there are no statistically significant differences between the M1: 5.0 g L-1 and M2: 7.5 g L-1 of dry yeast doses on the number of cobs per plant and the untreated plants (control). As well as between M2: 7.5 g L-1 and M3: 10.0 g L-1. p ( ) g g The results in Table 5 showed that the foliar application of active dry yeast had significantly augmented cobs weight plant-1 and plant weight of the aerial parts indicators compared with control (spraying with tap water). The maximum mean values have been recorded by applying 10 g L-1 of dry yeast in the average of both seasons 2019 and 2020, while the minimum values in the untreated control. Increasing the active dry yeast concentration from 0 to 10 g L-1 increased the cobs weight per plant and plant weight of the aerial parts. It is clear that the level of 10 g L-1 of dry yeast gave the highest record of studied plant weight of the aerial parts parameter 940.03 g, with an increase of 102.69% compared to the untreated plants as a control (recorded 463.77 g). 3.3. Chemical Characters at the Dough Stage It is seen in Table 6 that the phosphorus, potassium, calcium, magnesium, and iron concentrations have increased significantly as a result of the applied different dry yeast levels. The smallest contents of previous minerals were in the control plants, followed by the records obtained from applications of active dry yeast, which the highest values recorded with M3: 10 g L-1 yeast. M3 active yeast dose recorded increment by 47.05% in P, 61.73% in K, 92.67% in Ca, 51.42% in Mg, and 173.68% in Fe concentrations in comparison with non-treated plants as control. In this regard, Duru (2020) reported that calcium, sulfur, and potassium are essential mineral elements in human and animal nutrition. Here, it is known that P is a vital part of nucleic acids, phospholipids, adenosine triphosphate (ATP), and several coenzymes. Also, K affects osmosis and the operation of stomata by its role as a cofactor involved in protein formation, enzymes activation, and main solute functioning in water balance. Fast-growing animals seem to have higher potassium requirements, and an increased protein level increases the demands. Potassium is the main cation in intracellular fluid and actions in acidic medium, organization of osmotic pressure, conduction of the nerve impulse, muscle contraction, especially the heart muscle, cell membrane function, and Na+/K+-ATPase. Potassium is also required during glycogen synthesis. Like that, Ca is important in the formation and cell wall constancy and in the maintenance of membrane composition and permeability, the stimulation of certain enzymes coordinates many responses of cells to stimuli (Soetan et al., 2010). For animals, calcium acts as a component of bones and teeth and nerve and muscle function regulation. In blood coagulation, Ca activates the conversion of prothrombin to thrombin and also participates in the coagulation of milk. Calcium activates many enzymes such as ATPase, succinic dehydrogenase, lipase, etc. (Malhotra, 1998; Murray et al., 1999; Soetan et al., 2010). 315 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes In this regard, Mg is part of the chlorophyll particles; it activates many enzymes and an active substance of several enzyme systems in which thymine pyrophosphate is a cofactor. It also stimulates pyruvic acid carboxylase, pyruvic acid oxidase, and the condensing enzyme for the reactions in the Krebs cycle. 3.3. Chemical Characters at the Dough Stage It is also a component of bones and teeth. Fe is a component of cytochromes, electron transport, stimulates some enzymes, and plays a role in the formation of chlorophyll. Iron acts like hemoglobin in oxygen transportation. In cellular respiration, Fe acts as a major constituent of enzymes that contribute to biological oxidation, such as cytochrome C, C1, A1, etc. (Malhotra, 1998; Murray et al., 1999; Soetan et al., 2010). While the solubility of Iron, Fe2+, and Fe3+ ions decrease with an increase in pH being extremely low in calcareous soils, where noticeable Fe deficiency in plants not adapted to these conditions occurs (Römheld and Nikolic, 2006). Table 6. Effect of dry yeast foliar application on P, K, Ca, Mg, and Fe concentration (%) of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons Treatments P K Ca Mg Fe Spray Control 0.170±0.024 d 0.784±0.056 d 0.628±0.054 d 2.112±1.713 c 0.076±0.005 d M1: 5.0 g L-1 0.214±0.012 c 1.083±0.054 c 0.897±0.028 c 2.767±2.332 b 0.139±0.017 c M2: 7.5 g L-1 0.221±0.016 b 1.156±0.083 b 1.076±0.064 b 3.191±2.711 a 0.171±0.019 b M3: 10.0 g L-1 0.250±0.026 a 1.268±0.050 a 1.210±0.031 a 3.198±2.786 a 0.208±0.015 a Mean 0.214 1.073 0.953 2.817 0.149 P-Value < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 S.E.D.** 0.001333 0.00971 0.02393 0.0539 0.00317 The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. S.E.D.: Standard errors of differences of means. Table 6. Effect of dry yeast foliar application on P, K, Ca, Mg, and Fe concentration (%) of maiz under high pH soil conditions at the dough stage during the 2019 and 2020 seasons The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means. For micronutrients concentrations, it appeared from the results mentioned in Table 7 that active dry yeast applications increased the copper, selenium, and zinc contents of the samples statistically when compared to control plants. The highest Cu and Se amounts of the samples obtained from these applications were determined as 49.342 and 1.825 ppm for M3, respectively. In contrast, the highest Zn concentration resulted in M1 and M2, with non-significant changes in the copper concentration between them observed. 3.3. Chemical Characters at the Dough Stage Furthermore, the lowest concentrations detected by control with 27.190, 0.331, and 76.618 ppm for Cu, Se, and Zn, respectively. It was determined that there was an insignificant difference in the concentrations of Manganese elements between the yeast doses and the control (untreated plants). Copper is a component of vital redox and lignin-biosynthetic enzymes (Soetan et al., 2010), and it plays a role in iron intake (Chandra, 1990). Moreover, it is significant for growth and bone synthesis (Malhotra, 1998; Murray et al., 1999). Additionally, copper is required for the synthesis of collagen and elastin fibers that provide structure and elasticity to connective tissue and blood vessels (Larson, 2005). Selenium is integrated originally into plants by sulfur substitution in the cysteine and methionine (Soetan et al., 2010). Selenium is a mineral that is a component of a complete defense system that shelters the lively organism from the hurtful effect of reactive oxygen species (ROS) (Murray et al., 1999). Here Malhotra (1998) and Soetan et al. (2010) indicated that zinc is effective in the construction of chlorophyll, invigorates some enzymes, and functions as a part in the creation of auxin, chloroplasts, and starch. Moreover, the absorbed zinc that enters the liver was exported to peripheral tissue in plasma, bound to albumin after being united into zinc metalloenzymes. These results may be reflected that increasing the zinc concentration in the maize plants sprinkled with active dry yeast could lead to good results on animals' health and production. On the contrary, the use of dry yeast on corn has recorded a negative effect on molybdenum concentrations, where the control was the highest and gradually decreased with increasing the yeast doses from 5.0 to 10.0 g L-1. The Mo concentration was relatively high in corn silage and other feeds linked with the lowest Cu concentrations resulting in undesirable ratios between Cu and Mo (Miltimore and Mason, 1971). This may refer to the presence of an antithesis association between Cu and Mo concentrations in maize plants. 316 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Pur Table 7. Effect of dry yeast foliar application on Cu, Mn, Mo, Se, and Zn concentration (ppm) of maize grown under high pH soil conditions at the dough stage during the 2019 and 2020 seasons. 3.3. Chemical Characters at the Dough Stage The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means, NS: insignificant difference. Treatments Cu Mn Mo Se Zn Spray Control 27.190±2.73 d 116.048±27.99 NS 0.754±0.06 a 0.331±0.27 d 76.618±4.30 c M1: 5.0 g L-1 33.533±5.67 c 116.359±3.04 0.394±0.07 c 0.802±0.09 c 84.573±20.74 a M2: 7.5 g L-1 46.207±7.18 b 115.032±12.41 0.538±0.27 b 1.208±0.26 b 83.675±7.90 a M3: 10.0 g L-1 49.342±9.25 a 115.750±21.33 0.401±0.06 c 1.825±0.70 a 79.816±16.36 b Mean 39.07 115.80 0.52 1.04 81.17 P-Value < 0.001 0.908 < 0.001 < 0.001 < 0.001 S.E.D.** 0.3470 1.9010 0.002404 0.00765 0.5190 The means within each column within main effects and interactions followed by the same letter are not significant at P ≤ 5%. Mean±Standard deviation. *S.E.D.: Standard errors of differences of means, NS: insignificant difference. Similar results were in harmony with those of Abbas (2013) on beans, and Nasser et al. (2019) on lentils, they reported that chemical composition such as mineral elements improved under applying of yeast. On the other hand, Abbas (2013) revealed that the role of yeast could also be attributed to its effect on enzymatic activity, the production of some natural plant hormones, improving the ability to absorb nutrients, diversion of phosphorus from insoluble state to soluble one, and increasing its absorbability by plants, all of these increase the mineral content in plants. 4. Conclusion In the present study, foliar dry yeast applications to maize had a positive effect on the morphological and chemical properties, and as a result of these applications, the highest properties, including the amount of the studied properties, were obtained at a dose of 10 g L-1. The findings obtained from this study, when considered overall, it could be concluded that foliar applications of active dry yeast during the beginning vegetative stages might be recommended to increase vegetative growth, yield, and quality of maize for silage purposes in the soils that have high pH under Van-Turkey conditions. Furthermore, the fact that yeast is a natural and safe substance for the plants and the environment and economically feasible. Acknowledgments This study was supported by Scientific Research Projects (BAP) as project number FDK-2019- 8382. The authors would like to thank BAP for the big support. This study was supported by Scientific Research Projects (BAP) as project number FDK-2019- 8382. The authors would like to thank BAP for the big support. Abu Khouder, N. A., Abobaker, A. M. S., & Al-Mashat, K. M. E. (2019). Influence of some nitrogen fertilization and dry yeast extract levels on growth and pod yield of snap bean (Phaseolus vulgaris L.). Middle East Journal of Agriculture Research, 8(4), 973-982. g , , , pp Agamy, R., Hashem, M., & Alamri, S. (2013). Effect of soil amendment with yeasts as bio-fertilizers on the growth and productivity of sugar beet. African Journal of Agricultural Research, 8(1), 46-56. g ) f g , ( ), Acar, Z., Tan, M., Ayan, İ., Aşçi, Ö. Ö., Mut, H., Başaran, U., Gülümser, E., Can, M., & Kaymak, G. (2020). Türkiye’de yem bitkileri tarımının durumu ve geliştirme olanakları. Türkiye Ziraat Mühendisleri IX. Teknik Kongresi, 13-17 Ocak, Ankara, pp 529-554. Abed, A. M. R., & Zeboon, N.H. (2020). Effect of spraying with organic and bio-fertilizers on some growth characteristics of sunflower plant (Helianthus annuus L.). Plant Archives, 20 (2), 1050- 1055. References H., & Tawfik, M. M. (2011). Integrated effect of organic and biofertilizers on wheat productivity in new reclaimed sandy soil. Research Journal of Agriculture and Biological Sciences, 7(1), 105-114. Al- Ani, M. H. I., Al-Dulaimi, O. I. M., Mahmood, G. H. T., Al-Rawi, A. R. M., & Seadh, S. E. (2020). Response of some maize hybrids to foliar spraying treatments 1- growth characteristics. Systematic Reviews in Pharmacy, 11(11), 1672-1676. Al-madhagi, I. A. H. (2019). Effect of humic acid and yeast on the yield of greenhouse cucumber. Journal of Horticulture and Postharvest Research, 2(1), 67-82. Alsaady, M. H. M., Salim, H. A., Abdulrazzaq, A. K., Saleh, U. N., Jassim, N. H., Hamad, A. R., Attia, J. A., Darwish, J. J., & Hassan, A. F. (2020). Response of cabbage plants to foliar application of yeast suspension and nitrogen fertilizer. Ecology, Environment and Conservation, 26(2), 832- 836. Al-Shaheen, M. R., Asmail, M. H., & Al-Shaheen, M. R. (2019). Effect of iron nanoparticles and dry yeast extract on the yield and the productivity of corn (Zea mays L.). Iraqi Journal of Science, 60, 46-51. Altunlu, H., Demiral, O., Dursun, O., Sönmez, M., & Ergün, K. (2019). Effects of microbial fertilizer application on growth and yield of sweet corn (Zea mays L. var. saccharata). Atatürk University Journal of the Agricultural Faculty, 50(1), 32-39. Chandra, R. K. (1990). Micro-nutrients and immune functions: An overview. Annual New York Academy of Science, 587, 9-16. , C. E. (2020). Mineral and phytochemical evaluation of Zea mays Husk. Scientific African, 7, 1- Duru, C. E. (2020). Mineral and phytochemical evaluation of Zea mays Husk. Scientific African, 7, 1-8. El-Ghadban, E. A., Ghallab, A. M., & Abdel-Wahab, A. F. (2003). Effect of organic fertilizer and biofertilization on growth, yield and chemical composition of marjoram plants under newly reclaimed soil conditions. Journal Agriculture Science Mansoura, 28 (9), 6957-6973. EL-Kholy, M. H., Omar, M. N. A., & Salah, Z. S. (2007). Growth response of wheat crop to microbial inoculation and yeast strain under different mineral phosphorus fertilizer levels at Northeastern Delta of Egypt. Journal Agriculture Science Mansoura University, 32(2), 1513-1520. EzzEl-Din, A. A., & Hendawy, S.F. (2010). Effect of dry yeast and compost tea on growth and oil content of Borago officinalis plant. Research Journal of Agriculture and Biological Sciences, 6(4), 424-430. ( ) Ferguson, J. J., Avigne, W. T., Allen, L. H., & Koch, K. E. (1995). References Abbas, S. M. (2013). The influence of biostimulants on the growth and on the biochemical composition of Vicia faba Cv. Giza 3 beans. Romanian Biotechnological Letters, 18(2), 8061-8068. Abbas, S. M. (2013). The influence of biostimulants on the growth and on the biochemical composition of Vicia faba Cv. Giza 3 beans. Romanian Biotechnological Letters, 18(2), 8061-8068. Abed, A. M. R., & Zeboon, N.H. (2020). Effect of spraying with organic and bio-fertilizers on some growth characteristics of sunflower plant (Helianthus annuus L.). Plant Archives, 20 (2), 1050- 1055. Abed, A. M. R., & Zeboon, N.H. (2020). Effect of spraying with organic and bio-fertilizers on some growth characteristics of sunflower plant (Helianthus annuus L.). Plant Archives, 20 (2), 1050- 1055. Abu Khouder, N. A., Abobaker, A. M. S., & Al-Mashat, K. M. E. (2019). Influence of some nitrogen fertilization and dry yeast extract levels on growth and pod yield of snap bean (Phaseolus vulgaris L.). Middle East Journal of Agriculture Research, 8(4), 973-982. Abu Khouder, N. A., Abobaker, A. M. S., & Al-Mashat, K. M. E. (2019). Influence of some nitrogen fertilization and dry yeast extract levels on growth and pod yield of snap bean (Phaseolus vulgaris L.). Middle East Journal of Agriculture Research, 8(4), 973-982. g ) f g ( ) Acar, Z., Tan, M., Ayan, İ., Aşçi, Ö. Ö., Mut, H., Başaran, U., Gülümser, E., Can, M., & Kaymak, G. (2020). Türkiye’de yem bitkileri tarımının durumu ve geliştirme olanakları. Türkiye Ziraat Mühendisleri IX. Teknik Kongresi, 13-17 Ocak, Ankara, pp 529-554. Acar, Z., Tan, M., Ayan, İ., Aşçi, Ö. Ö., Mut, H., Başaran, U., Gülümser, E., Can, M., & Kaymak, G. (2020). Türkiye’de yem bitkileri tarımının durumu ve geliştirme olanakları. Türkiye Ziraat Mühendisleri IX. Teknik Kongresi, 13-17 Ocak, Ankara, pp 529-554. g , , , pp Agamy, R., Hashem, M., & Alamri, S. (2013). Effect of soil amendment with yeasts as bio-fertilizers on the growth and productivity of sugar beet. African Journal of Agricultural Research, 8(1), 46-56. 317 YYU J AGR SCI 32 (2): 310-319 YYU J AGR SCI 32 (2): 310-319 Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Purposes ( ) Alobaidy et al. / Determination Effects of Active Dry Yeast on Morphological and Chemical Components of Maize Plants Grown in Alkaline Soils for Silage Pur Ahmed, M. A., Ahmed, G., Mohamed, M. References Growth of CO2 enriched sour orange seedlings treated with gibberellic and cytokinins. Florida State Horticultural Society, 99, 37- 39. Gomez, K. A., & Gomez, A. A. (1984). Statistical Procedures for Agricultural Research. 2nd Edition, John Wiley and Sons, New York, 680 p. Hassan, N. M. K., Marzouk, N. 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Study the effect of dry bread yeast Saccharomyces cerevisiae, in some qualities of vegetative growth and nutritional status of the Lens culinaris. Plant Archives, 19(2), 1101-1106. Öz, A., Kapar, H., & Dok, M. (2017). Mısır Tarımı. Karadeniz Tarımsal Araştırma Enstitüsü Yayınları, pp 5. [Accessed:15.09.2020, http://arastirma.tarim.gov.tr/ktae/Belgeler/brosurler/Mısır%20Tarımı.pdf]. Römheld, V., & Nikolic, M. (2006). Iron. In: Barker, A.V., Pilbeam, D.J. (Eds.), Handbook of Plant Nutrition. CRC Press, Boca Raton, FL, pp. 329-350. Rouf Shah, T., Prasad, K., & Kumar, P. (2016). Maize - A potential source of human nutrition and health: A review. Cogent Food & Agriculture, 2(1), 1-9. Sabancı, İ. (2015). Mısır-soya birlikte üretim şekillerinin bazı agronomik özelliklere etkisi. (Doktora tezi), Adnan Menderes Üniversitesi, Ziraat Fakültesi, Aydın. Saleh, S., Liu, G., Liu, M., Ji, Y., He, H., & Gruda, N. (2018). 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Estimation levels of Indole acetic acid (IAA) and gibberellic acid (GA3) in dry bakery yeast Saccharomyces cereviciae. Journal of Bio-Technology Research Center, 4 (2), 94- 100. Vas, P. R., & Papanas, N. (2019). Editorial and Mini-Review: Topical oxygen therapy for diabetic foot ulcerations-avenue towards new hope? The Review of Diabetic Studies, 15, 71-73. Wanas, A. L. (2006). Trails for improving growth and productivity of tomato plants grown in winter. Annuals Agricultural Science Moshtohor, 44(3), 466-471. Xi, Q., Lai, W., Cui, Y., Wu, H., & Zhao, T. (2019). Effect of yeast extract on seedling growth promotion and soil improvement in afforestation in a semiarid chestnut soil area. Forests, 10(76), 1-16. 319	17,884
hal-01882636-samplepaper.txt_2	French-Science-Pile	Open Science	Various open science	2,018	Evolving Living Technologies-Insights from the EvoEvo project. SSBSE 2018 - 10th Symposium on Search-Based Software Engineering, Sep 2018, Montpellier, France. pp.46-62, ⟨10.1007/978-3-319-99241-9_2⟩. ⟨hal-01882636⟩	None	English	Spoken	1,016	2,000	5 Conclusion As the 2011 FET Consultation Report “Living Technology, Artificial Systems, Embodied Evolution” shows, many approaches have been proposed to create living technologies. Now at the end of the EvoEvo project, and having created what we think is a living technology (“EvoMove”), one can draw the big picture of the design principles we identified and briefly exposed here. Indeed, we claim that the key insight into building evolutionary living technologies is to go back to a fundamental property of living systems. Living systems are in essence strongly integrated systems, while technological systems are, by construction, strongly modular systems. Evolutionary living technologies will only be efficient if they are strongly integrated within the systems (in order to enable the system to innovate) and with their users, be it a real person (as in EvoMove), or a software entity (as in commensal architecture). In some sense, this proposal is not a total surprise, since it is a similar mind shift as the one that happened at the end of the 1980s in robotics. The development of Behavior-Based Robotics under the impulsion of Rodney Brooks was nothing else than the close integration of robots with their environments and of robots components one with the others [5]. We now propose that software systems themselves, although they are not physical entities, follow the same path 16 G. Beslon et al. in order to be able to leverage Darwinian evolution to dynamically react and adapt to their users. This will enable living software systems to co-construct their behavior with a user who, in that same moment, will become a partner of this behavior. Acknowledgments This work was supported by the European Commission 7th Framework Program (FP7-ICT-2013.9.6 FET Proactive: Evolving Living Technologies) EvoEvo project (ICT- 610427, http://www.evovo.eu/). The authors thank all the partners of the EvoEvo project for fruitful discussions. References 1. Abernot J., Beslon G., Hickinbotham S., Peignier S., Rigotti, C. (2017) Evolving Instrument Based on Symbiont-Host Metaphor: a Commensal Computation. Journal of Creative Music Systems, 2(1):1–10. 2. Banzhaf W., Baumgaertner B., Beslon G., Doursat R., Foster J. A., McMullin B., Veloso de Melo V., Miconi T., Spector L., Stepney S., White R. (2016) Defining and Simulating Open-Ended Novelty: Requirements, Guidelines, and Challenges. Theory in Biosciences, 135(3):131–161 3. Batut B., Parsons D.P., Fischer S., Beslon G., Knibbe C. (2013) In silico experimental evolution: a tool to test evolutionary scenarios. BMC bioinformatics, 14(15):S11 4. Beslon G., Liard V., Elena S.F. (2016) Evolvability drives innovation in viral genomes. In:2nd EvoEvo Workshop, satellite workshop of CCS2016, Amsterdam, September 2016, 6 p. 5. Brooks R.A., (1990) Elephants don’t play chess. Robotics and autonomous systems, 6(1-2):3–15. 6. Chatterjee K., Pavlogiannis A., Adlam B., Nowak M.A. (2014) The time scale of evolutionary innovation. PLoS computational biology, 10(9):e1003818 7. Clark E.B., Hickinbotham S.J., Stepney S. (2017) Semantic closure demonstrated by the evolution of a universal constructor architecture in an artificial chemistry Journal of the Royal Society Interface, 14:20161033 8. Colizzi E.S., Hogeweg P. (2014) Evolution of functional diversification within quasispecies. Genome biology and evolution, 6(8):1990–2007 9. Colizzi E.S., Hogeweg, P. (2016) High cost enhances cooperation through the interplay between evolution and self-organisation. BMC evolutionary biology, 16(1):31 10. Cuypers T.D., Hogeweg P. (2012) Virtual genomes in flux: an interplay of neutrality and adaptability explains genome expansion and streamlining. Genome Biology and Evolution, 4(3):212–229 11. de Boer F.K., Hogeweg P. (2012) Co-evolution and ecosystem based problem solving. Ecological informatics 9:47–58 12. https://youtu.be/p eJFiQfW1E 13. Fischer S., Bernard S., Beslon G., Knibbe C. (2014) A model for genome size evolution. Bulletin of mathematical biology, 76(9): 2249–2291 14. Hickinbotham S., Stepney S. (2016) Bio-reflective architectures for evolutionary innovation. ALife 2016, Cancun, Mexico, pp.192–199. MIT Press. Evolving Living Technologies — Insights from the EvoEvo project 17 15. Hickinbotham S., Stepney S. (2016) Augmenting live coding with evolved patterns. EvoMusArt, Porto, Portugal, LNCS 9596:31–46. Springer. 16. Hindré T., Knibbe C., Beslon G., Schneider D. (2012) New insights into bacterial adaptation through in vivo and in silico experimental evolution, Nature Reviews Microbiology, 10:352–365. 17. Hooper L. V., Gordon J. I. (2001). Commensal host-bacterial relationships in the gut. Science, 292:1115–1118. 18. Hooper L. V., Midtvedt T., Gordon J. I. (2002) How host-microbial interactions shape the nutrient environment of the mammalian intestine. Annual review of nutrition, 22(1):283–307 19. Hoverd T., Stepney S. (2016) EvoMachina: a novel evolutionary algorithm inspired by bacterial genome reorganisation, 2nd EvoEvo Workshop, CCS 2016, Amsterdam, Netherlands. 20. Kimura M. (1968) Evolutionary Rate at the Molecular Level. Nature, 217:624–626 21. Kriegel H.-P., Krger P., Zimek A. (2009) Clustering highdimensional data: A survey on subspace clustering, pattern-based clustering, and correlation clustering. ACM Transactions on Knowledge Discovery from Data, 3(1):1–58 22. Pagie L., Hogeweg P. (1997) Evolutionary consequences of coevolving targets. Evolutionary computation, 5(4):401–418 23. Patrikainen A., Meila M. (2006) Comparing subspace clusterings. IEEE Transactions on Knowledge and Data Engineering, 18(7):902–916 24. Peignier S., Rigotti C., Beslon, G. (2015) Subspace clustering using evolvable genome structure. In Proceedings of the 2015 Annual Conference on Genetic and Evolutionary Computation, pp. 575–582 25. Peignier S., Abernot J., Rigotti C., Beslon, G. (2017) EvoMove: Evolutionary-based living musical companion. In European Conference on Artificial Life (ECAL), pp. 340–347 26. Peignier S., Rigotti C., Rossi A., Beslon G. (2018) Weight-based search to find clusters around medians in subspaces. ACM Symposium on Applied Computing. p.10. 27. Plucain J., Hindr T., Le Gac M., Tenaillon O., Cruveiller S., Mdigue C., Leiby N., Harcombe W. R., Marx C. J., Lenski R. E., Schneider D. (2014) Epistasis and allele specificity in the emergence of a stable polymorphism in Escherichia coli. Science, 343:1366–1369 28. Rocabert C., Knibbe C., Consuegra J., Schneider D., Beslon G. (2017) Beware batch culture: Seasonality and niche construction predicted to favor bacterial adaptive diversification. PLoS computational biology, 13(3):e1005459 29. Rutten J., Hogeweg P., Beslon G. (in prep) Adapting the engine to the fuel: mutator populations can reduce the mutational load by reorganizing their genome structure. in prep. 30. Szathmry E., Maynard-Smith J. (1997) The major evolutionary transitions, Nature, 374(6519):227–232 31. van Dijk B., Hogeweg P. (2016) In silico gene-level evolution explains microbial population diversity through differential gene mobility. Genome biology and evolution, 8(1):176–188.	12,659
W2228763662.txt_1	German-Science-Pile	Open Science	Various open science	null	None	None	German	Spoken	2,561	5,874	DOI 10.5162/12dss2015/P1.2 Ansätze für die Kalibrierung und die quantitative Messung mit Halbleitergassensoren im sub-ppm Bereich 1 1 2 1 Caroline Schultealbert , Andreas Schütze , Stefan Böttcher und Tilman Sauerwald 1 Lehrstuhl für Messtechnik, Universität des Saarlandes, Saarbrücken 2 Pharmazeutische und Medizinische Chemie, Universität des Saarlandes, Saarbrücken, c.schultealbert@lmt.uni-saarland.de Abstract: Eine Methode zur kalibrierten quantitativen Messung von Spurengasen wird vorgestellt und am Beispiel von Toluol getestet. Dafür wird ein modellbasierter Ansatz zur Verbesserung der quantitativen Messung mittels eines speziellen temperaturzyklischen Betriebs (engl. temperature cycled operation, TCO) genutzt. Außerdem werden transportable Kalibrierstandards entwickelt. Die Kalibrierstandards nutzen die gasförmige Phase über einem Toluol-Squalan-Gemisch im thermodynamischen Gleichgewicht. Mit reinem Squalan lässt sich eine Nullprobe erzeugen für eine absolute und hintergrundunabhängige Quantifizierung durch Differenzbildung. Über ein einfaches Modell kann die Konzentration im Gas aus dem Mischungsverhältnis abgeschätzt werden. Die Abschätzung wurde für die hergestellten Kalibrierstandards mit Hilfe von quantitativen Vergleichsmessungen mittels GC-MS bestätigt. Die Sensorsignale für die Kalibrierstandards wurden mit Messungen an einer Gasmischanlage verglichen und zeigen die Vergleichbarkeit beider Methoden. Mit der neuen Methode konnte ein Sensor (AS-MLV) für eine quantitative Messung sowohl in Nulluft als auch in ungereinigter Laborluft in einem Konzentrationsbereich von 100 ppb bis 100 ppm kalibriert werden. Keywords: Halbleitergassensor, quantitative Messung, Kalibrierung, Raumluft-Überwachung, Flüchtige organische Komponenten (VOCs) Einleitung Mit Hilfe von Halbleitergassensoren können flüchtige organische Komponenten (engl. volatile organic compounds, VOCs) auch in Konzentrationen von wenigen ppb detektiert werden [1–2]. Aus diesem Grund werden sie unter anderem für die Überwachung von Raumluft eingesetzt [2]. Für jede Anwendung, die eine quantitative Messung solcher Stoffe benötigt, ist eine rückführbare Kalibrierung Voraussetzung. Im Labor können Sensoren mit Hilfe von Gasmischanlagen (GMA) kalibriert werden [3–4]. Da dieses Verfahren skalierbar ist, ist es auch für die Werkskalibrierung von Sensoren in großer Stückzahl geeignet. Für eine Kalibrierung einzelner Sensoren am Einsatzort ist die Methode wegen des hohen Aufwands allerdings nicht geeignet. Die hier vorgestellte Methode bietet die Möglichkeit Kalibrierstandards beliebiger Konzentrationen verschiedener Gase (in diesem Fall Toluol) dem Sensor vor Ort anzubieten. Grundlage bildet ein thermodynamisches Gleichgewicht in einem abgeschlossenen Volumen bestehend aus einer flüssigen und einer gasförmigen Phase. Voraussetzung für eine erfolgreiche Kalibrierung ist ein stabiles Messsignal der 12. Dresdner Sensor-Symposium 2015 Sensoren. Stabilität und Drift gehören zu den größten Herausforderungen in Hinblick auf die erfolgreiche Anwendung von Halbleitergassensoren in der Praxis [5–6]. Die Quantifizierung basiert auf der Geschwindigkeit Relaxation der Sauerstoffbedeckung der Sensoroberfläche nachdem der Sensor in einen Ungleichgewichtszustand versetzt wurde [7]. Erreicht wird dieser Ungleichgewichtszustand durch die Variation der Sensortemperatur. Dieser sogenannte temperaturzyklische Betrieb (engl. temperature cycled operation, TCO) erhöht sowohl die Stabilität [8] als auch die Sensitivität [7] und die Selektivität des Sensors [2]. Quantifizierungsmethode Die Quantifizierungsmethode beruht auf der Ratenkonstante der Reaktion der in der Luft vorhanden reduzierenden Gase mit dem auf der Sensoroberfläche adsorbierten Sauerstoff. Das zu Grunde liegende Modell und Ergebnisse zur Quantifizierung von Ethanol und Benzol sind in [7] ausführlich dargestellt, weswegen an dieser Stelle nur kurz darauf eingegangen wird. Für die Bestimmung der Ratenkonstante wird der Sensor zunächst in einen Zustand mit hoher Sauerstoffbedeckung (hohe Temperatur, 106 DOI 10.5162/12dss2015/P1.2 hier 450 °C) versetzt. Wird ein Temperatursprung durchgeführt, durchläuft der Sensorleitwert zunächst ein Minimum und relaxiert dann auf einen Gleichgewichtswert. Die Geschwindigkeit dieser Relaxation, bei der umgebende Gase mit dem ionosorbierten Sauerstoff der Oberfläche reagieren, ist unter anderem abhängig von der Konzentration und Art dieser Gase. Berechnet wird die Ratenkonstante � über zwei Parameter: der Energiebarriere �� , die sich wiederum aus der Differenz der logarithmierten Leitwerte direkt vor und nach dem Temperatursprung errechnet �� = �� ln � �� (1) und der Steigung des logarithmischen Leitwerts innerhalb des Relaxationsplateaus: � � �= � ∙ �� (�) ��. (2) Experimentelles Für die beschriebene Form der Bestimmung der Ratenkonstante, wird ein Sensor mit einer geringen thermischen Zeitkonstante benötigt. Aus diesem Grund wird ein AS-MLV Sensor verwendet, ein auf SnO2 basierender Sensor der Firma ams AG. Die sensitive Schicht ist auf einer mikromechanischen Membran aufgebracht, was ein schnelles Heizen und Abkühlen mit einer thermischen Zeitkonstante von unter 10 ms ermöglicht [7]. Der Temperaturzyklus setzt sich aus zwei Phasen zusammen: einer Oxidationsphase bei 450 °C über 300 ms und einer Relaxationsphase bei einer niedrigeren Temperatur über 200 ms. Insgesamt werden zehn Temperaturen zwischen 200 °C und 380 °C mit einem Abstand von 20 °C für die Relaxation ausgewählt. Dies ergibt einen 5 s Zyklus, bestehend aus 20 verschiedenen Temperatursprüngen, dargestellt in Abbildung 1. Aus den 10 Sprüngen auf niedrige Temperatur können 10 Ratenkonstanten nach Formel (2) bestimmt werden. Temperatursteuerung und Leitwertmessung werden durch ein SniffChecker-Modul der Firma 3S GmbH – Sensors, Signal Processing, Systems realisiert, dessen Messbereichs mit einem Logarithmierverstärker LOG112 von Texas Instruments Inc. erweitert wurde [7]. Das geschlossene Volumen für die Erzeugung der Kalibrierstandards wird durch eine 50 ml Duran Protect Laborflasche dargestellt. Der Sensor wird für die Messung in einem Deckel montiert und auf die Laborflasche geschraubt. 12. Dresdner Sensor-Symposium 2015 Abbildung 1: Temperaturzyklus bestehend aus 10 Relaxationsund Oxidationsplateaus und zugehörigem Sensorleitwert. Konzept der Kalibrierung Die für die Kalibrierung benötigten Gase werden durch ein thermodynamisches Gleichgewicht in einem geschlossenen Volumen bestehend aus einer flüssigen und einer gasförmigen Phase hergestellt. Die Konzentration einer Komponente in der Dampfphase wird bestimmt durch ihren Partialdruck �� . Bei einer reinen Substanz ist der Partialdruck im thermodynamischen Gleichgewicht gleich dem Sättigungsdampfdruck �� der Flüssigkeit [9]. Besteht die Flüssigkeit im betrachteten System aus mehreren Komponenten verändert sich ihr Dampfdruck in Abhängigkeit vom Stoffmengenanteil � in der Lösung. Für verdünnte Lösungen wird der Dampfdruck durch das Henry-Gesetz �� = � ∙ �� beschrieben. Die Henry-Konstante �� ist für jede Kombination von Stoffen spezifisch anzugeben und stellt einen virtuellen Dampfdruck dar [9]. Bei Kenntnis der Henry-Konstante ist es damit möglich beliebige Konzentrationen für die Kalibrierung von Gassensoren herzustellen. Eine Abschätzung der Konstante kann mit Hilfe des UNIFAC-Modells erzielt werden[10]. Dieses im Vergleich zu Monte-Carlo-Simulationen einfach anzuwendende Modell ist eine sogenannte Gruppenbeitragsmethode, bei der die chemische Aktivität (Abweichung von der Idealität innerhalb von Mischungen) des Stoffs anhand der vorhandenen funktionellen Gruppen bestimmt wird. Für das Stoffpaar Toluol in Squalan wird daraus eine Henry-Konstante von 1,9 kPa bei 20 °C gewonnen, was deutlich niedriger als der Sättigungsdampfdruck von reinem Toluol ist (2,9 kPa) und daher eine gute Mischbarkeit beider Flüssigkeiten impliziert. Aufgrund des starken Abstiegs des Dampfdrucks und damit auch der HenryKonstante mit der Temperatur [12] muss diese sehr genau erfasst werden. 107 DOI 10.5162/12dss2015/P1.2 Die Grundsubstanz Squalan wurde vor allem aufgrund ihres vernachlässigbaren Dampfdrucks (0,02 µPa bei Raumtemperatur [11]) ausgewählt, um mögliche Quereinflüsse auf die Messung zu verhindern. Vor der Verwendung wird die Substanz bei erhöhter Temperatur (100 °C) über 72 h mit Stickstoff durchspült. Dadurch werden flüchtige Verunreinigungen entfernt. Die besonders hohe Reinheit der Matrix führt auch dazu, dass andere in der Raumluft vorhandene Stoffe (z.B. VOCs) während der Messung in ihr gelöst werden, bis sich ein thermodynamisches Gleichgewicht einstellt. Dadurch wird ihre Konzentration während der Kalibrierung deutlich reduziert, Für VOC ist der Verteilkoeffizient (d. h. der Quotient aus Gasteilchen pro Volumen in Luft bzw. in Squalan) deutlich kleiner als eins (für Toluol ca. -4 10 ). Die Konzentration von VOC wird daher typischerweise um einige Größenordnungen reduziert. Auf diese Weise kann mit einem reinen Squalan-Standard ein nahezu VOCfreies Gas erzeugt werden und damit ein Hintergrundsignal unter sonst gleichen Umgebungsbedingungen, wie Temperatur, Feuchte und Permanentgaskonzentration, aufgenommen werden. Um einen großen Bereich von Konzentrationen abzudecken und im Besonderen niedrige Konzentrationen mit hoher Genauigkeit herzustellen wird eine Verdünnungsreihe durchgeführt. Auf diese Weise können Kalibrierproben mit Konzentrationen bis in den ppb-Bereich hergestellt werden. Aus der Abschätzung der Einwaagefehler kann ein Fehler von maximal 5 % für alle Proben in der Verdünnungsreihe errechnet werden. Dies ist für die meisten Anwendung wie der Quantifizierung von VOCs in Raumluft ausreichend. Für alle Proben wird der Dampfdruck von Toluol mit Hilfe des UNIFAC-Modells bei 20°C (Raumtemperatur) bestimmt. Mit Hilfe eines GC-MS von Thermo Fisher Scientific Inc. wird dazu die Konzentration im Dampfraum von ToluolSqualan Mischungen gemessen (siehe Tabelle 1). Die Quantifizierung der GC-MS Messungen erfolgte mittels eines internen Standards und die Kalibrierung anhand eines reinen Toluol-Headspaces. Die GC-MS Messungen zeigen, dass die Bestimmung der HenryKonstante mit dem UNIFAC-Modell gut geeignet ist, insbesondere die Proben 355 ppm und 3520 ppm Toluol in Squalan zeigen eine gute Übereintimmung. Für hohe und niedrige Stoffmengen können Abweichungen aufgrund der begrenzten Gültigkeit des Henry-Gesetzes (hohe Stoffmengen) und Ungenauigkeiten in der durchgeführten Ein-Punkt-Kalibrierung des 12. Dresdner Sensor-Symposium 2015 GC-MS (niedrige Stoffmengen) zu Fehlern führen. Tabelle 1: Aufstellung der hergestellten Proben bestehend aus Toluol verdünnt in Squalan zur Erzeugung von Konzentrationen zwischen 3,13 ppm und 11,7 ppb bei einer Raumtemperatur von 20 °C (Werte erhalten aus dem UNIFAC Modell und den GC-MS Messungen). UNIFAC �� [ppm] �� [ppm] 20 °C 35100 GC-MS �� [ppm] 25 °C �� [ppm] 20 °C 866 1860 1440 3520 86,8 112 86,4 355 9,25 10,6 8,19 166 3,13 58,9 1,11 39,7 0,98 3,23 2,50 17,3 0,325 6,11 0,114 1,84 0,0344 0,63 0,0117 Ergebnisse Für die Charakterisierung der Quantifizierung wurden Messungen an einer Gasmischanlage durchgeführt. Die Erzeugung der gewünschten Toluol-Konzentrationen erfolgte mit einem Permeationsofen. Der quasistatische Leitwert am Ende des 450 °C Plateaus reagiert stark auf das Einschalten des Gases, erreicht ein Maximum und relaxiert dann über mehrere Minuten auf einen niedrigeren Wert (siehe Abbildung 2 am Beispiel von 1 ppm Toluol). Abbildung 2: Reaktion des Sensors auf ein Gasangebot von 1 ppm Toluol beginnend bei � = 0. Der quasistatische Sensorleitwert am Ende des 450 °C Plateaus schwingt stark über, was eine Quantifizierung unmöglich macht, die 108 DOI 10.5162/12dss2015/P1.2 Ratenkonstante erreicht mit Einsetzen des Gases einen konstanten Wert. Hintergründen, und die zugehörige Hintergrundmessung mit Hilfe einer Squalanprobe. Durch dieses überschwingende Verhalten ist eine Quantifizierung nicht gut möglich. Der Verlauf der berechneten Ratenkonstante, der ebenfalls in Abbildung 2 zu sehen ist, erreicht mit Beginn des Gasangebots einen konstanten Wert und ermöglicht so eine sofortige Quantifizierung ohne Einlaufen. VOCs in der Raumluft ist ein Konzentrationsbereich im ppb-Bereich von Interesse. In Abbildung 4 sind die Ratenkonstanten aller 10 Relaxationstemperaturen für Toluol-Konzentrationen zwischen 10 ppb und 3 ppm zu sehen. Aufgenommen wurden diese Daten mit Hilfe der Kalibrierstandards aus Tabelle 1. Eine aufgenommene Kalibrierkurve sollte im Idealfall unabhängig von den vorherrschenden Umgebungsbedingungen gültig sein. Dies ist aufgrund der hier vorgestellten Methode der Kompensation des Hintergrunds möglich. Zu sehen ist dies in Abbildung 5. Die dort enthaltenen Messungen wurden über einen weiten Konzentrationsbereich unter völlig unterschiedlichen Bedingungen aufgenommen. Die schwarzen Messpunkte stellen Messungen an der Gasmischanlage dar, der Hintergrund ist dort nahezu rein und muss somit nicht abgezogen werden. Die roten Punkte wurden von Messungen mit den hier vorgestellten Kalibrierstandards erhalten und wurden damit in einem normalen Raum aufgenommen. Der Hintergrund zur Differenzbildung wurde durch eine Probe mit gereinigtem Squalan erzeugt. Trotz dieser sehr stark unterschiedlichen Bedingungen liegen alle Messungen näherungsweise auf einer Kurve. Aufgrund einiger vor allem messtechnischer Probleme (Messbereich & Abtastrate) sind insbesondere die Randbereiche fehlerbelastet und der derzeit als für die Quantifizierung und Kalibrierung zuverlässig zu erachtende Bereich liegt zwischen 100 ppb und 100 ppm. Die Durchführung einer solchen Hintergrundmessung ist, wie bereits erwähnt, durch das Angebot einer Probenkammer mit gereinigtem Squalan möglich, da dieses bei der Einstellung des thermodynamischen Gleichgewichts die meisten flüchtigen Verunreinigungen aus der Luft entfernt. Abbildung 3 zeigt beispielhaft zwei Zyklen, beide bei ca. 1 ppm Toluol, aufgenommen unter stark unterschiedlichen Umgebungsbedingungen, innerhalb der Gasmischanlage vor Nullluft, einem nahezu leeren Hintergrund, und innerhalb einer Kalibrierprobe vor Raumluft. Der Zyklus der Hintergrundmessung in einer squalangefüllten Kalibrierprobe ist ebenfalls dargestellt. Der Mittelwert der Ratenkonstanten �� des an der GMA aufgenommenen Zyklus beträgt 1,5 ∙ � 10�� , die durch Differenz gebildete mittlere � Rate der Kalibrierprobe bei einer leicht höheren � Konzentration 1,8 ∙ 10�� . Es zeigt sich, dass � trotz der hohen Verschiebung des Sensorleitwerts zwischen Raumluft und Gasmischer die Ratenkonstante für die Messung sehr nahe zusammen liegen. Unter der Annahme einer linearen Abhängigkeit von Ratenkonstante und Konzentration beträgt die Abweichung der Messungen weniger als 10 %. Innerhalb einer Größenordnung ist die Annahme eines linearen Zusammenhangs gerechtfertigt. Für größere Konzentrationsbereiche, folgt das Messsignal einem Potenzgesetz. Für die Überwachung von Diskussion & Ausblick Die Herstellung von transportablen Kalibrierstandards mit der beschrieben Methode ist sehr vielversprechend. Mit dem UNIFAC-Model lassen sich für beliebige VOC schnell die gewünschten Ratenkonstanten abschätzen. Abbildung 3: Zyklen zweier Messungen bei ca. 1 ppm Toluol, aufgenommen vor unterschiedlichen 12. Dresdner Sensor-Symposium 2015 109 DOI 10.5162/12dss2015/P1.2 Halbleitergassensoren Laborbedingungen. außerhalb definierter Quellen [1] ELMI, I. ; ZAMPOLLI, S. ; COZZANI, E. ; MANCARELLA, F. ; CARDINALI, G. C.: Development of ultra-low-power consumption MOX sensors with ppb-level VOC detection capabilities for emerging applications. In: Sensors and Actuators, B: Chemical 135 (2008), S. 342– 351 Abbildung 4: Die Ratenkonstante verschiedener Temperatursprünge über der Konzentration in doppelt logarithmischer Darstellung. [2] LEIDINGER, M. ; SAUERWALD, T. ; REIMRINGER, W. ; VENTURA, G. ; SCHÜTZE, A.: Selective detection of hazardous VOCs for indoor air quality applications using a virtual gas sensor array. In: Journal of Sensors and Sensor Systems 3 (2014), S. 253–263 [3] HELWIG, N. ; SCHÜLER, M. ; BUR, C. ; SCHÜTZE, A. ; SAUERWALD, T.: Gas mixing apparatus for automated gas sensor characterization. In: Measurement Science and Technology 25 (2014). [4] ENDRES, H.-E. ; JANDER, H. D. ; GÖTTLER, W.: A test system for gas sensors. In: Sensors and Actuators B: Chemical 23 (1995), S. 163– 172 [5] BARSAN, N. ; KOZIEJ, D. ; WEIMAR, U.: Metal oxide-based gas sensor research: How to? In: Sensors and Actuators, B: Chemical 121 (2007), S. 18–35 Abbildung 5: Messpunkte verschiedener Konzentrationen aufgenommen an der GMA und mit Hilfe der hergestellten Kalibrierproben und abgezogenem Hintergrund. Trotz der stark unterschiedlichen vorherrschenden Umgebungsbedingungen liefern beide Messungen ähnlliche Ergebnisse, sodass eine unabhängige Kalibrierkurve angegeben werden kann. Eine Rückmessung kann mit GC-MS erfolgen. Die notwendigen Voraussetzungen für eine Kalibration vor Ort sind damit gegeben. Eine Herausforderung stellt die Ungenauigkeit der Sensormessung selbst dar: statische Sensorsignale zeigen hier eine sehr starke Drift und auch eine starke Abhängigkeit vom Gashintergrund und sind daher schlecht geeignet. Deutlich besser ist in diesem Punkt das neu untersuchte Merkmal der Ratenkonstante. Ein erheblicher Nachteil dieses Merkmals ist aber die starke Rauschbelastung, da hier Signale verrechnet werden, die bei sehr kleinen Strömen (im Bereich von einigen nA) gemessen wurden. Dies ist vermutlich ein Grund für die in Abbildung 5 dargestellte starke Streuung der Messpunkte; eine weitere Verbesserung der Messwerterfassung ist daher ein wichtiges Ziel. Trotz der Schwächen ist dies ein vielversprechender Umgebungsbedingungen eine zur kalibrierten quantitativen Messung mit 12. Dresdner Sensor-Symposium 2015 [6] KOROTCENKOW, G. ; CHO, B. K.: Instability of metal oxide-based conductometric gas sensors and approaches to stability improvement (short survey). In: Sensors and Actuators, B: Chemical 156 (2011), S. 527–538 [7] BAUR, T. ; SCHÜTZE, A. ; SAUERWALD, T.: Optimierung des temperaturzyklischen Betriebs von Halbleitergassensoren. In: Technisches Messen 82 (2015), S. 187–195 [8] REIMANN, R. ; SCHÜTZE, A.: Fire detection in coal mines based on semiconductor gas sensors. In: Sensor Review 32 (2012), S. 47–58 [9] ATKINS, P.: Physikalische Chemie. WILEY-VCH Verlag GmbH, 2001 [10] FREDENSLUND, A. ; JONES, R. L. ; PRAUSNITZ, J. M.: Group-contribution estimation of activity coefficients in nonideal liquid mixtures. In: AIChE Journal 21 (1975), Nr. 6, S. 1086–1099 [11] PIACENTE, V. ; FONTANA, D. ; SCARDALA, P.: Enthalpies of Vaporization of a Homologous Series of n-Alkanes Determined from Vapor Pressure Measurements. In: Journal of Chemical & Engineering Data 39 (1994), S. 231–237 [12] SMITH, F. L. ; HARVEY, A. H.: Avoid common pitfalls when using Henry ’s law. In: Chemical Engineering Progress September (2007), S. 33– 39 110.	6,770
hal-02179699-RAlg.txt_3	French-Science-Pile	Open Science	Various open science	2,021	Real spectra and ℓ-spectra of algebras and vector lattices over countable fields. Journal of Pure and Applied Algebra, In press, ⟨10.1016/j.jpaa.2021.106861⟩. ⟨hal-02179699v4⟩	None	Unknown	Unknown	1,192	2,503	Therefore, π ◦ ρ is a surjective measure from Vk onto D, so it defines a surjective homomorphism from Idℓc Vk onto D. By Stone duality, (1) follows. Now let us prove (2). We may assume that k is a real-closed field. Replacing the real field R by k in the argument of the proof, relative to R[[G]], of Dickmann et al. [13, Proposition 1.1], then taking G a divisible totally ordered Abelian group with exactly two nonzero Archimedean classes, we get a totally ordered real-closed k-algebra A, without zero divisors, such that the real spectrum of A is the twoelement chain. Denoting by K any real-closed field extension of A, it follows that the map ε : 3 ։ 2, defined by ε(0) = 0 and ε(1) = ε(2) = 1, is represented, with respect to the Stone dual of the functor Specr , by the inclusion map from A into K. Therefore, mimicking part of the argument of [34, Theorem 5.4], Spec D ∼ = Specr R 26 F. WEHRUNG where the “condensate” R is defined as the k-algebra of all eventually constant sequences of elements of K with limit in A. Finally, there is no least x ∈ D such that (ω, 2) ≤ (ω, 1) ∨ x; whence D is not a generalized dual Heyting algebra. Using Corollary 10.7, Item (3) follows. This suggests the following problem. Problem. Characterize the spectral spaces that can be represented as real spectra of algebras over any formally real field. References [1] Kirby A. Baker, Free vector lattices, Canad. J. Math. 20 (1968), 58–66. MR 0224524 [2] Elı́as Baro, Normal triangulations in o-minimal structures, J. Symbolic Logic 75 (2010), no. 1, 275–288. 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Algebra 224 (2020), no. 4, 106202. MR 4021916 [37] , From non-commutative diagrams to anti-elementary classes, J. Math. Logic 21 (2021), no. 2, 2150011. MR 4290500. [38] Günter M. Ziegler, Lectures on Polytopes, Graduate Texts in Mathematics, vol. 152, SpringerVerlag, New York, 1995. MR 1311028 Normandie Université, UNICAEN, CNRS UMR 6139, LMNO, 14000 Caen, France Email address: friedrich.wehrung01@unicaen.fr URL: https://wehrungf.users.lmno.cnrs.fr.	24,798
2013STRAJ052_6	French-Science-Pile	Open Science	Various open science	2,013	Développement de nouveaux nanovecteurs pour les thérapies anti-HCV/HCC	None	English	Spoken	7,408	14,010	RESULTS AND DISCUSSION Formation of BrIBAM-HSA Capsules. Protein film growth through HSA adsorption and refunctionalization with BrIBAM moieties on nonporous silica (SiO2) particles were followed by flow cytometry. Fluorescently labeled HSA-rhodamine isothiocyanate (RITC) was adsorbed for 30 min onto BrIBAM-modified SiO2 particles (5 μm diameter) to generate the first protein coating. The protein surface was then functionalized with BrIBAM groups by the addition of an excess of 2-bromoisobutyryl bromide, which reacts with the pendent amine groups of the lysine or arginine residues of the protein, converting them into BrIBAM moieties. Subsequently, a second HSA coating was deposited over 30 min by adsorption onto the BrIBAM-functionalized HSA surface. This coating process was repeated up to five times to form a film denoted (BrIBAM-HSA)5. Fluorescence intensity data obtained from flow cytometry (Figure 1a) illustrate the near-linear and continuous growth of the protein film on SiO2 particles with increasing number of HSA coating steps. The initial fluorescence intensity of SiO2 particles (26 ( 1 au) after the first HSA coating step (n = 1) increased by ca. 23 au increments per protein coating step until a final MERTZ ET AL. Figure 2. Fluorescence microscopy images of BrIBAM-HSA capsules initially templated onto 5 μm diameter SiO2 particles after (a) n = 2 and (b) n = 5 HSA-RITC coating steps. Insets show bright field microscopy images of the capsules. AFM images of the BrIBAM-HSA capsules initially templated onto 3 μm diameter SiO2 particles in the dried state after (c) n = 2 and (d) n = 5 HSA coating steps. (e) Evolution of the capsule wall thickness as a function of the number of HSA coating steps, as determined from AFM z-profile analysis. Scale bars are 10 μm for fluorescence microscopy images, 5 μm for bright field microscopy images, and 2 μm for AFM images. fluorescence intensity of 119 ( 5 au was achieved after the fifth HSA coating step (n = 5). Flow cytometry experiments of control particles produced using the same conditions, but without intermittent functionalization of the surface with BrIBAM groups, revealed no significant increase in fluorescence intensity (Figure 1a), thus confirming the key role of the BrIBAM moieties in driving the assembly and growth of the protein film. Fluorescence microscopy images (Figure 1b) of the protein-coated particles after each HSA coating step reveal regular protein deposition onto the SiO2 particles, as shown by the increase in the fluorescence of the particles with each coating step. Removal of the underlying SiO2 template from the BrIBAM-HSA coated particles after each coating step (n = 2!5) yielded homogeneous and well-dispersed BrIBAMprotein hollow capsules with negligible shrinkage (see Figure 2a,b for fluorescence and bright field microscopy images of capsules after n = 2 VOL . 6 ’ NO. 9 ’ 7584–7594 ’ 7586 2012 www.acsnano.org ARTICLE Figure 3. Disassembly of BrIBAM-HSA capsules imaged by fluorescence microscopy. (BrIBAM-HSA)3 capsules (a) dispersed in water, (b) incubated with acetone (ca. 1 min), and (c) incubated with urea (3 M aqueous solution) (ca. 4 h). Scale bars are 10 μm. and 5 coating steps). Atomic force microscopy (AFM) images (Figure 2c,d) of the protein capsules in the dried state for n = 2 and 5 HSA coating steps show homogeneous structures with significantly higher densities compared with our previously reported capsules prepared by a single HSA adsorption step onto BrIBAM modified surfaces.42 The capsule wall thickness with HSA coating step (Figure 2e), as determined by AFM z-profile analysis (Supporting Information, Figure S1), linearly increases, which is consistent with the flow cytometry data and fluorescence microscopy images (Figure 1). The wall thickness of the protein capsules ranged from ca. 6 ( 2 nm for the first HSA coating to ca. 14 ( 3 nm after five HSA coatings, with an average thickness increment of ca. 2 nm per HSA coating (for n = 1!5). Uniform film growth is attributed to a regular and consistent functionalization of the protein lysine or arginine groups with BrIBAM moieties after each HSA coating step. Previously, we demonstrated that BrIBAM interactions with proteins and other biopolymers occur through noncovalent interactions and that both the isobutyramide and bromine substituted BrIBAM moieties play a role in the cohesive process.42 Therefore, the noncovalent nature of the sequential BrIBAM-HSA capsules was investigated by assessing their intermolecular disassembly. (BrIBAM-HSA)3 capsules were incubated with solutions of either acetone or urea (3 M aqueous solutions). In both cases, complete disassembly was observed with protein release into solution occurring rapidly with acetone (<1 min) and more slowly with urea (ca. 4 h) (Figure 3). These results are similar to those we previously reported for BrIBAM-HSA capsules obtained with one HSA coating step, confirming that the interactions between HSA and BrIBAM-functionalized HSA are noncovalent in nature and are readily disrupted acetone or urea. BrIBAM versus IBAM. The role of the bromine atoms attached to the isobutyramide groups in the assembly of the protein films was experimentally assessed by using the bromine-free IBAM groups. Thus, the same procedure as that described for the BrIBAM studies was employed, with the exception that isobutyryl chloride was used in place of 2-bromoisobutyryl bromide. Initially, HSA was adsorbed onto IBAM-functionalized SiO2 MERTZ ET AL. particles and then the lysine and arginine groups of the surface-coated protein were refunctionalized with IBAM groups via reaction with isobutyryl chloride followed by further protein adsorption. The protein film growth via HSA adsorption and IBAM refunctionalization was followed by flow cytometry (Figure 4a). Similar to the BrIBAM system, a near-linear increase in fluorescence intensity with increasing number of HSA coating steps was monitored. However, comparison of the fluorescence intensity increments obtained per coating step from flow cytometry for the BrIBAM and IBAM systems (performed with the same acquisition parameters) indicates a significantly lower slope for the IBAM system (ca. 13 au) compared to the BrIBAM system (ca. 23 au). Over five coating steps, the fluorescence intensity of the (IBAM-HSA)5 films (78 ( 4 au) was ca. 30% less than that for the (BrIBAM-HSA)5 films. These results suggest that the IBAM approach is suitable for the assembly and growth of protein films, although significantly less protein is adsorbed (30% less after five coating steps) compared with the BrIBAM approach. Removal of the SiO2 templates from the IBAMHSA coated particles for n = 2 and 5 yielded homogeneous and well-dispersed HSA hollow capsules with negligible shrinkage (Figure 4b,c and Supporting Information, Figure S2). AFM images recorded in the dried state (Figure 4e,f) show homogeneous capsules (for n = 2 and 5 HSA coating steps). AFM z-profile analysis (Supporting Information, Figure S3) revealed thinner capsule walls (see thickness graph in Figure 4d) for the IBAM-HSA capsules (5.5 ( 2 nm for n = 2, 10 ( 2 nm for n = 5) compared to their BrIBAM-HSA analogues (7 ( 3 nm for n = 2, 14 ( 3 nm for n = 5) (Figure 2c,d). Interestingly, when only one HSA coating is assembled on IBAM-modified SiO2 particles, capsules are not observed (Figure S4). In contrast, two HSA coatings yield (IBAM-HSA)2 capsules and may constitute a suitable alternative to BrIBAM-HSA capsules when the presence of bromine atoms is not desired. IBAMmediated binding most likely results from a dense network of hydrogen bonds between the IBAM moieties and the peptide backbone of the proteins. The flow cytometry results and AFM analysis clearly emphasize the simultaneous roles of the amide and bromide functionalities to form noncovalent VOL. 6 ’ NO. 9 ’ 7584–7594 ’ 7587 2012 www.acsnano.org ARTICLE Figure 4. Growth of IBAM-HSA films and capsules. (a) Evolution of the fluorescence intensity with increasing number of HSARITC coating steps on SiO2 particles, as followed by flow cytometry. Fluorescence microscopy images of IBAM-HSA capsules initially templated onto 5 μm diameter SiO2 particles after (b) n = 2 and (c) n = 5 HSA-RITC coating steps. (d) Evolution of the capsule wall thickness as a function of the number of HSA coating steps, as determined from AFM z-profile analysis. AFM images of IBAM-HSA capsules initially templated onto 3 μm diameter SiO2 particles in the dried state after (e) n = 2 and (f) n = 5 HSA coating steps. Scale bars are 10 μm for fluorescence microscopy images and 2 μm for AFM images. interactions generated within the BrIBAM-HSA and IBAM-HSA assemblies. To assess the wettability of the BrIBAM and IBAM graf ts, water contact angle measurements of silicon slides graf ted with BrIBAM and IBAM moieties were performed. Similar values of 52 ( 3! and 56 ( 1! were measured for the BrIBAM and IBAM films, respectively, whereas blank silicon slides (control experiments) possessed a lower contact angle value of 22 ( 2! ( Support ing Information, Figure S5). These results demonstrate that BrIBAM and IBAM grafts have a similar and quite significant hydrophobicity, and that the Br atom from the BrIBAM moiety does not increase the hydrophobicity of the films. Noncovalent Interactions. Recently, the implication of the larger halogen atoms (Cl, Br, and I) in the formation of halogen bonds that are able to compete with hydrogen bonds in biological systems has been assessed.43!50 Here, we calculated the electrostatic potential surfaces of bromine and hydrogen atoms by using the Hartree!Fock quantum model (Figure 5a,b) for the BrIBAM and IBAM compounds. A positive value of the electrostatic potential (50 kJ mol!1) calculated at the top of the bromine atom reveals an electron-deficient region (referred to as a “sigma-hole”) that is able to interact favorably with electron donating groups such as oxygen atom lone pairs characteristic of carbonyl groups.43 In comparison, alkyl hydrogen atoms, such as the methine proton from IBAM, are not able to establish strong bonding interactions with electron donors. This suggests that the bromine extremity of BrIBAM behaves as a Lewis acid that may interact with electron donors present in the HSA proteins. Moreover, such MERTZ ET AL. atomic charge distribution is very similar to that obtained with models of halogenated compounds forming halogen bonds with proteins. Numerous electron donor atoms are present in the polypeptide chains of proteins (N, O, and S) and the most common halogenbonds encountered between proteins complexed with halogenated compounds involve the oxygen atoms on the protein carbonyl groups.43,47!49 A recent X-ray crystallography study on dibromo-amide modified taxane bound to tubulin revealed a short and highly directed Br!O contact (3.17 Å, angle C!Br 3 3 3 O of 173!) that was identified as a halogen-bond.50 Figure 5c shows the electrostatic potential surfaces of the dibromo-amide modified taxane that were mapped via the same Hartree!Fock model used above. Comparison of the electrostatic potential surfaces for the bromine atom of the dibromo-amide modified taxane forming a halogen-bond with BrIBAM indicates very similar atomic charge distributions and values of electrostatic potential (ca. 51 kJ mol!1 calculated at the top of the bromine). As a result of the theoretical insights obtained through these calculations and data gathered from the literature, we postulate that bromine atoms play a role, through halogen-bonding, in the formation of the thicker BrIBAM-HSA films (∼30%) when compared with IBAM-HSA films. Biofunctionalization. Biofunctionalization of the BrIBAM-HSA and IBAM-HSA protein films with other biological macromolecules after subsequent reinitiation with BrIBAM or IBAM grafts was investigated with fluorescently labeled alkaline phosphatase (RITClabeled, AP-RITC), dextran (fluorescein isothiocyanatelabeled, DEX-FITC) and DNA plasmid (stained with VOL. 6 ’ NO. 9 ’ 7584–7594 ’ 7588 2012 www.acsnano. org ARTI CLE Figure 5. Chemical structures (Lewis and 3D) and ab initio electrostatic potential surfaces of (a) an IBAM moiety, (b) a BrIBAM moiety, and (c) a dibromo-amide modified taxane establishing an halogen-bond within tubulin (3.17 Å, angle C!Br 3 3 3 O of 173!).45 The C!H (IBAM) and equivalent C!Br (BrIBAM) bonds are shown perpendicular to the page. The C!Br bonds of the dibromo-amide modified taxane are also shown perpendicular to the page and the Br establishing a halogen-bond is represented in the front. R represents a part of the brominated taxane molecule distant from the halogen bond. The potential energies are presented in the!75 to þ75 kJ mol!1 range to emphasize the variation in electrostatic potential associated with the halogen atom. The gray arrows point to positive electrostatic potentials at the apex of the bromine atoms and are indicative of the presence of an electron-deficient region (also referred to as a sigma-hole) that are able to establish favorable interactions with electron donating groups such as the lone pairs of carbonyl groups. green fluorescent YOYO-1, a high affinity nucleic acid stain, DNAp-YOYO). Flow cytometry experiments performed on SiO2 templates (Figure 6a) coated with (BrIBAM-HSA)4 or (IBAM-HSA)4 protein films indicate that the three biopolymers AP, DEX, and DNAp adsorbed onto the reinitiated protein surface. Moreover, significantly less biopolymer was adsorbed when coated onto IBAM surfaces compared to BrIBAM surfaces (ca. 30, 50, and 35% less for AP, DEX, and DNAp, respectively). These results are consistent with the presence of a higher amount of the BrIBAM grafts (compared to IBAM) resulting from a higher protein quantity and thickness, as observed above. Better retention of the biopolymers due to possible interactions with the bromine atoms may also explain this effect. Figure 6b,d show the resulting (BrIBAM-HSA)4 capsules functionalized with AP, DEX, and DNAp, respectively. Similar biofunctionalized IBAM-HSA capsules were also obtained (Supporting Information, Figure S6). Retention of the biological functionality of the adsorbed outer layer was assessed using APfunctionalized BrIBAM-HSA capsules. The continuous and linear conversion of p-nitrophenylphosphate (PNP) into p-nitrophenol (PN) catalyzed by the APfunctionalized BrIBAM-HSA capsules and followed in solution by UV/Vis spectrophotometry (at λ = 410 nm, maximum absorption of PN) demonstrates that the absorbed AP enzymes retain their activity (Figure 6e). Control experiments without any AP coating indicated that neither BrIBAM grafts nor HSA protein promoted MERTZ ET AL. the conversion of PNP into PN. The enzymatic activity of the equivalent amount of alkaline phosphatase with that contained in the AP-functionalized BrIBAM-HSA capsules was also investigated in bulk solution (see details in Supporting Information, Figure S7). The reaction kinetics were found to be considerably faster (ca. 150 times) compared with the AP-functionalized BrIBAM-HSA capsules. This effect may be explained by the fact that for the free enzyme in bulk solution, the enzymatic sites are more accessible compared to enzymes and HSA associated within the protein capsule wall. Cytotoxicity Studies. Smaller SiO2 particle templates (1.11 and 0.585 μm diameter) were employed for the formation of HSA capsules with more suitable sizes for biomedical applications and cellular interactions36,51,52 (i.e., e 1 μm). The same process as described previously was employed to prepare (BrIBAM-HSA)4 and (IBAMHSA)4 capsules. After template removal, the capsules were well-defined and readily dispersed in aqueous buffer, thus demonstrating the versatility of the process to tune the size of the capsules. Transmission electronic microscopy (TEM) images of these capsules in the dried state confirmed homogeneous structures with diameters of ca. 1 and 0.5 μm for both BrIBAMHSA (Figure 7a,b) and IBAM-HSA (Figure 7c,d) capsules. To assess the effect of these BrIBAM-HSA and IBAMHSA capsules on cell viability, an MTT assay was conducted by incubating various doses of the two types of HSA capsules (with sizes of 1 and 0.5 μm) with HeLa cells (Figure 7e). The results demonstrate that even at a VOL . 6 ’ NO. 9 ’ 7584–7594 ’ 7589 2012 www.acsnano.org ARTI CLE Figure 6. Biofunctionalization of BrIBAM-HSA and IBAM-HSA protein films with biological macromolecules. (a) Comparison followed by flow cytometry of the relative adsorption content of AP, DEX, and DNAp adsorbed onto (BrIBAM-HSA)4-BrIBAM and (IBAM-HSA)4-IBAM protein films coated onto silica templates. Fluorescence intensities are normalized for clarity. Fluorescence microscopy images of (BrIBAM-HSA)4-BrIBAM capsules functionalized with (b) AP-RITC, (c) DEX-FITC, and (d) DNAp-YOYO. Scale bars are 8 μm. (e) Kinetic plot of the conversion of PNP into PN and a phosphate ion catalyzed by the (BrIBAM-HSA)4-(BrIBAM-AP) capsules and followed by UV/Vis spectrophotometry at 410 nm. Figure 7. TEM images of (BrIBAM-HSA)4 capsules with diameters of (a) 1 and (b) 0.5 μm. TEM images of (IBAM-HSA)4 capsules with diameters of (c) 1 and (d) 0.5 μm. Scale bars are 0.5 μm. (e) Cell viability of HeLa cells incubated with (IBAM-HSA)4 (0.5 and 1 μm diameter) and (BrIBAM-HSA)4 (0.5 and 1 μm diameter) capsules at different capsule/cell ratios (100:1 and 10:1) for 24 h, as measured by an MTT assay. Cell viability is normalized as a percentage of untreated cells. The data are with the mean and standard error of two independent experiments, each performed in triplicate. high dose (ca. 100 capsules per cell), the cell viability and metabolic activity after an incubation period of 24 h are not significantly affected by the capsules (compared with untreated cells), suggesting no significant cytotoxicity. Following this initial study on cell viability in the presence of the capsules, further investigations in vitro and in vivo will be required to fully assess the biocompatibility of these capsules. Cellular Association with BrIBAM-HSA Capsules. We next investigated the effect of capsule wall thickness on cellular association. AlexaFluor (AF) 488-labeled BrIBAM-HSA capsules (1 μm in diameter) with different MERTZ ET AL. HSA layer numbers (from 2 to 5) were incubated with HeLa cells at a capsule-to-cell ratio of 100:1. The extent of cellular association at 6 and 24 h was evaluated using flow cytometry. It was shown that approximately 32% and 63% of the cells were associated with (BrIBAM-HSA)2 capsules after incubation for 6 and 24 h, respectively, which is significantly higher than the other multilayered (BrIBAM-HSA)n capsules with n = 3!5 for the same time periods (Figure 8a). The higher extent of (BrIBAM-HSA)2 capsule internalization was confirmed by deconvolution fluorescence microscopy imaging. Microscopy images show that significantly VOL. 6 ’ NO. 9 ’ 7584–7594 ’ 7590 2012 www.acsnano.org Figure 8. Interaction of AF488-labeled BrIBAM-HSA capsules composed of different layer numbers with HeLa cells for 6 and 24 h incubation at 37!C. (a) Percentage of cells associated with capsules was quantified by flow cytometry. Data are the mean ( standard deviation. At least 5000 cells were analyzed in each experiment. Representative images of cells internalized with (b) (BrIBAM-HSA)2 or (c) (BrIBAM-HSA)5 capsules are shown. more (BrIBAM-HSA)2 capsules (Figure 8b) were internalized in HeLa cells compared with (BrIBAM-HSA)5 capsules (Figure 8c). Previous studies have demonstrated that the internalization of hollow capsules involves capsule deformation. 53,54 Furthermore, Fery et al. reported that the stiffness of polyelectrolyte capsule shells is proportional to the square of the shell thickness.55,56 We postulate that the (BrIBAMHSA)2 capsules are the most flexible among the series of HSA capsules studied, favoring deformation and resulting in enhanced cellular uptake. Taken together, these results show that the sequential assembly of HSA capsules via BrIBAM grafts affords control over cellular uptake of the capsules. This opens the way for possibilities in biomedical applications, such as targeted drug delivery, where the balance between improved biodistribution and high rate of drug administration are closely linked with cellular uptake. METHODS Materials. 3-Aminopropyltriethoxysilane (APTS), ethanol (EtOH), ammonium hydroxide solution (28 wt % in water), isobutyryl chloride (IBC), 2-bromoisobutyryl bromide (BrIB), triethylamine (Et3N), N,N-dimethylformamide (DMF), rhodamine B isothiocyanate (RITC), sodium bicarbonate (NaHCO3), dimethylsulfoxide (DMSO), acetone, urea, hydrofluoric acid (HF, 48 wt % in H2O), ammonium fluoride, poly(ethylene imine) (PEI), human serum albumin (HSA), alkaline phosphatase (AP) from bovine intestinal mucosa, p-nitrophenylphosphate (PNP), and FITC labeled dextran (FITC-DEX) (Mw = 70 kDa) were obtained from Sigma-Aldrich and used as received. DNA plasmid (DNAp) was provided by A.W. Püschel (Münster Universität, Germany) (pBK-ha-NRP1). Nonporous silica particles of various sizes (5 wt % suspensions, average diameters of 5.35, 3.25, 1.11, and 0.585 μm) were obtained from Microparticles GmbH (Berlin, Germany). YOYO-1 iodide DNA-stain, Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), Alexa Fluor 488 carboxylic acid, succinimidyl ester (AF488), Dulbecco's phosphate-buffered saline (DPBS), and (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide) (MTT) were purchased from Invitrogen and used as received. MERTZ ET AL. The preparation of sequentially assembled noncovalent protein films based on the physical adsorption of HSA and chemical refunctionalization with isobutyramide grafts (i.e., isobutyramide or bromoisobutyramide) was reported. This approach allows for the formation of protein films with controlled growth profiles onto silica particles that, after template removal, yield hollow protein capsules with controllable wall thicknesses. These protein capsules displayed no significant cytotoxicity toward HeLa cells and their capsule wall thickness was found to influence their cellular uptake of HSA capsules, suggesting that the sequential assembly approach has significant potential for tailored cell studies. Versatile biofunctionalization of such protein capsules was demonstrated with the efficient formation of AP-, DEX-, and DNA plasmid-coated protein capsules that maintain biofunctionality, as demonstrated with AP-functionalized BrIBAM-HSA capsules. On the basis of the differences observed for the BrIBAM-HSA and IBAM-HSA systems, the driving force for protein assembly was hypothesized to result from a network of hydrogenand halogen-bonding interactions arising from the amide groups and bromine atoms, respectively. Understanding the processes associated with the cohesion of such protein assemblies is of primary importance to develop smart bioresponsive materials. Current investigations are focused on elucidating the local structure of the protein films, including the degree of organization and electronic environment at the molecular/atomic level around the BrIBAM and IBAM moieties. In addition, functionalization of the capsules with cell-targeting biomolecules (e.g., sugars or aptamers) and therapeutic drugs is under investigation. ARTICLE CONCLUSION Fluorescent Labeling of HSA and AP. HSA was fluorescently labeled with RITC using a HSA:RITC mole ratio of 1:2. RITC was dissolved in DMSO (10 mg mL!1) and gently mixed with a solution of HSA (10 mg mL!1) dissolved in 0.1 M NaHCO3 buffer (pH 8.5) for 1 h. An identical procedure was employed for APRITC. HSA-RITC and AP-RITC were purified by size exclusion chromatography (SEC). The SEC column was equilibrated with Milli-Q water (20 mL) and then loaded with the concentrated protein solutions. The purified HSA-RITC and AP-RITC were then collected by eluting with Milli-Q water. For cell association experiments, HSA was labeled with AF488. Briefly, 30 mg of HSA was dissolved in 50 mM PBS buffer (pH 7.4). The mixture was stirred for 3 h and dialyzed against Milli-Q water to purify the AF488-labeled HSA. The labeled HSA was freeze-dried and stored at 4!C before use. Bromoisobutyramide (BrIBAM)and Isobutyramide (IBAM)-Modified Silica Particles. Nonporous silica particles (200 μL, 5 wt % in EtOH) were dispersed in a solution of EtOH (500 μL) containing ammonium hydroxide solution (30 μL, 30 wt % in water) and APTS (125 μL). After 2 h, the amino-modified particles were isolated via centrifugation and washed with EtOH (1 " 200 μL) and DMF (3 " 200 μL). A solution containing BrIB (65 μL) in anhydrous DMF(400 μL) was added dropwise to the amino-modified silica VOL. 6 ’ NO. 9 ’ 7584–7594 ’ 7591 2012 www.acsnano.org MERTZ ET AL. calculated from two independent experiments performed in triplicate. Cellular Association Experiments. HeLa cells were plated at 7.5! 104 cells/well in 24-well plates. AF488-labeled HSA capsules with various layers of HSA (from 2 to 5 layers) were added to the cells at a capsule-to-cell ratio of 100:1. The cells were incubated with the capsules at 37!C for 6 and 24 h. After the treatment, the cells were washed with DPBS three times, trypsinized, and resuspended in DPBS. The cellular association was analyzed by flow cytometry. Experiments were performed in duplicate, and at least 5000 cells were analyzed in each experiment. Characterization Methods. Flow Cytometry. A Cyflow Space (Partec GmbH) flow cytometer using an excitation wavelength of 488 nm (emission filter of 560"590 nm) was used to follow the growth of the BrIBAM-HSA-RITC and IBAM-HSA-RITC films on silica particles in Milli-Q water (pH 5.8). Flow cytometry graphs show evolution of fluorescence intensity (au) with the number of HSA coating steps. At least 20000 particles were analyzed in each experiment. A Miltenyi flow cytometer using an excitation wavelength of 488 nm (emission filters of 490"520 nm and 560" 590 nm) was used to compare the relative amount of fluorescently labeled AP-RITC, DEX-FITC, and YOYO-1 stained DNAp onto (BrIBAM-HSA)4 and (IBAM-HSA)4 coated silica particles. At least 20000 particles were analyzed in each experiment. Bright Field and Fluorescence Microscopy. Images of the protein-coated silica particles and protein capsules were taken using an Olympus IX71 inverted fluorescence microscope equipped with a DIC slider (U-DICT, Olympus), the corresponding filter sets, and a 60! oil immersion objective (Olympus UPFL20/0.5NA, W.D. 1.6). Atomic Force Microscopy (AFM). The imaged protein capsules were deposited onto PEI-coated silicon wafers, rinsed extensively with Milli-Q water, and allowed to air-dry. AFM scans of the capsules were carried out with a JPK NanoWizard2 BioAFM and with a D3000 Nanoscope IIIa (Veeco, Santa Barbara, CA). Typical scans were conducted in intermittent contact mode (AC mode) with silicon cantilevers (NSC/CSC) (MikroMasch, Bulgaria) using the accompanying JPK image processing software and in contact mode with a silicon nitride tip (model MSCT-AUHW, Veeco, Santa Barbara, CA) using the accompanying Veeco processing software. Contact Angle Measurements. Contact angle was measured on a Digidrop (Romans, France) apparatus. Three droplets of pure water (2 μL) were deposited and analyzed for each sample. UV/Vis Spectrophotometry. A SAFAS (Monaco) spectrophotometer was used to measure evolution of the absorbance as a function of the time of p-nitrophenol (PN) formation catalyzed by the (BrIBAM-HSA)4-BrIBAM-AP capsules in NaHCO3 buffer solution (pH 8.5, 0.05 M). Transmission Electron Microcopy (TEM). HSA capsules were deposited onto Formvar-coated copper grids, rinsed extensively with Milli-Q water, and allowed to dry in air. TEM analysis was carried out with a Philips CM120 BioTWIN instrument operated at 120 kV. Theoretical Study: Mapping of Electrostatic Potential Surface (Hartree " Fock Model). The method ology used here is similar to that used in a study describing the importance of halogen bonds in biological systems.48 Molecular models of the BrIBAM and IBAM moieties were constructed and their geometries optimized at the 3-21G() Hartree"Fock level (Figure 5). Electrostatic potential surfaces were generated by mapping the 3-21G() electrostatic potentials onto surfaces of molecular electron density (0.002 electron per Å3) by using the SPARTAN program (Wave function, Irvine, CA). Note that the 3-21G() basis set was used because the 6-31G() basis set and above are incomplete for bromine atoms. The limitations related to the use of electrostatic potential surfaces are discussed at length by Mecozzi et al. who demonstrated their usefulness in providing quantitative guidelines for evaluating the ability of aromatic compounds to form cation"π interactions.58 Conflict of Interest : The authors declare no competing financial interest . ARTICLE particles previously dispersed in a solution of DMF (400 μL) and Et3N (40 μL). After 2 h, Milli-Q water (200 μL) was added to dissolve the formed precipitates, and the resulting particles were isolated via centrifugation and washed with DMF (3! 200 μL). The same procedure was used to prepare IBAMmodified silica particles with the exception that IBC (55 μL) was used in place of BrIB. Protein Coating and Re functional ization Processes. BrIBAMand IBAM-modified silica particles dispersed in DMF (100 μL, 5 wt %) were added to aqueous solutions of HSA-RITC (1.0 mL, 0.25 mg mL"1) and allowed to stand for 30 min, followed by centrifugation and washing with Milli-Q water (1 mL). The resulting BrIBAM-HSA and IBAM-HSA coated particles (n = 1 coating step) were washed with anhydrous DMF (3! 100 μL) and redispersed in a solution of DMF (200 μL) and Et3N (20 μL). Separately, BrIB (32 μL) and IBC (27 μL) were dissolved in DMF (200 μL) and slowly added to the previously dispersed BrIBAMHSA and IBAM-HSA coated particles, respectively. After 2 h, MilliQ water (100 μL) was added to dissolve the formed precipitates and the resulting particles were isolated via centrifugation and washed with DMF (3! 100 μL). A second HSA coating step was performed by addition of the particles to a fresh solution of HSARITC (1.0 mL, 0.25 mg mL"1) in Milli-Q water for 30 min followed by centrifugation and washing with Milli-Q water (1 mL). After analysis, these HSA-coated particles (n = 2 coating steps) were refunctionalized with BrIBAM and IBAM moieties and HSA coating was repeated according to the above procedure to afford particles with up to five HSA coatings (n = 5). Biofunctionalization. (BrIBAM-HSA)4 coated particles were first refunctionalized with BrIBAM moieties via the protocol described above and subsequently biofunctionalized by the adsorption (1 h) of AP-RITC (0.5 mg mL"1), DEX-FITC (0.5 mg mL"1), or DNAp (0.1 mg mL"1) in Milli-Q water (1 mL) followed by centrifugation and washing with Milli-Q water (1 mL). YOYO-1 aqueous solution (10000 fold dilution) as a green fluorescent DNA stain was added for 20 min to the (BrIBAM-HSA)4-BrIBAM-DNAp coated particles (0.1 wt %) followed by particle washing with Milli-Q water. (IBAMHSA)4 coated particles were refunctionalized with IBAM moieties and biofunctionalized via the same procedure. Preparation of HSA and HSA-biopolymer (AP, DEX, or DNA) Capsules. Caution! This method utilized HF which is highly toxic and great care must be taken when handling. For one HSA coating (n = 1), the BrIBAM-HSA coated particles (1 μL, 0.5 wt %) were deposited onto a PEI-coated slide (glass, silicon, or gold wafer) and brought into contact with ammonium fluoride (8 M) buffered HF (2 M) (NH4F/HF, 1 μL, pH 5) for 1 min, followed by extensive rinsing of the slide with MilliQ water. For n g 2 HSA coating steps (n = 2"5), BrIBAM-HSA coated particles (100 μL, 1 wt %) and IBAM-HSA coated particles (100 μL, 1 wt %) were added to ammonium fluoride (250 μL, 8 M) buffered HF (2 M, pH 4) and mixed very gently for 2 min. The resulting capsules were then centrifuged at low speed (1800" 2000 g) for 15 min and washed with Milli-Q water (3! 50 μL). The process is identical to that used for the HSA-biopolymer capsules. Measurement of Enzymatic Activity of HSA -AP Capsule s . Enzymatic activity of (BrIBAM-HSA)4-BrIBAM-AP capsules was detected by using p-nitrophenylphosphate (PNP). The PNP solution (1 mL) at a concentration of 1.7 mg mL"1 in 0.05 M NaHCO3 buffer (pH = 8.5) was brought in contact with the (BrIBAM-HSA)4-BrIBAM-AP capsules and the subsequent formation of p-nitrophenol (PN) and a phosphate ion by enzymatic conversion of PNP was followed by UV/Vis spectrophotometry at λ = 410 nm (maximum absorption). Cell Viability Experiments. HeLa cells were cultured in DMEM media containing 10% FBS at 37!C in a humidified atmosphere containing 5% CO2. Cell viability was measured by reduction of MTT, as described previously.57 Briefly, HeLa cells were seeded at 5! 103 cells/well in 96-well plates and incubated with capsules at various ratios in 200 μL of growth media for 24 h. After treatment, cells were further incubated with fresh medium containing MTT (0.5 mg mL"1) for 4 h. The resulting blue formazan was solubilized using 150 μL of acidified isopropyl alcohol (0.04 N HCl), and the absorbance at 570 nm was measured with a plate reader (Multiskan Ascent, Thermo Scientific). The cell viability of treated cells was normalized as a percentage of untreated cells. The mean ( the standard errors were Acknowledgment. This work was supported by the Australian Research Council under the Discovery (F.C.) and Federation VOL. 6 ’ NO. 9 ’ 7584–7594 ’ 7592 2012 www.acsnano.org Supporting Information Available: AFM images and z-profile analysis of BrIBAM-HSA and IBAM-HSA capsules, bright field images of IBAM-HSA capsules, contact angle measurements, fluorescence microscopy images of IBAM-HSA biofunctionalized capsule s with AP, DEX , and DNAp and enzymatic activity of alkaline phosphatase in solution . This material is available free of charge via the Internet at http://pubs.acs.org. REFEREN CES AND NOTES 1. Vörös, J. 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Habersetzer,! Michel! Doffoël,! Pieter! Leyssen,! Johan! Neyts,! Mirjam! B.! Zeisel! and! Thomas! F.! Baumert.!(Manuscrit)soumis)à)Hepatology)! Nous! avons ! souhaité! étudier! l’ efficacité ! antivir ale! a insi! que ! la ! toxicité! d’ inhibiteurs ! de ! l’entrée! du! HCV! (tels ! que! CD81,! SR"BI,! et! CLDN"1)! en! combina ison! avec! différentes! molécules! antivirales! (IFN"! α,! DAA,! HTA)! in#vitro.! Pour! cela,! nous! avons! réalisé! un! screening! d’un! grand! nombre! de! combinaisons! de! ces! molécules! et! d’anticorps! ciblant! les! inhibiteurs! d’entrée,! mettant! en! évidence! la! présence! de! synergie! d’inhibition! marquée! entre! certaines! molécules! à! différentes!concentrations!sans!toxicité!remarquable.! C es!résultats! s uggèrent!le! développement ! de!nouvelles!stratégi es !antivirales!basées! sur !ces!sy nergie s.! Ma)participation):)réalisation!d’une!partie des!expériences!de!combinaisons,!maîtrise!et! mise! en! place! de! séquences! d’actions! à! l’aide! d’un! logiciel! pour! programmer! un! robot! afin! d’automatiser!une!partie!du!screening.!!!!!!! "!167!"! Synergy of entry inhibitors with direct-acting antivirals uncovers novel combinations for prevention and treatment of hepatitis C Isabel Fofana1,2, Fei Xiao1,2, Christine Thumann1,2, Roxane Alles1,2,3, Laurent Mailly1,2, Eric Robinet1,2, Nicolas Meyer4, Mickaël Schaeffer4, François Habersetzer1,2,5, Michel Doffoël1,2,5, Pieter Leyssen6, Johan Neyts6, Mirjam B. Zeisel 1,2 and Thomas F . Baumert 1,2,5 1 Inserm, U1110, Strasbourg, France, 2University of Strasbourg, Strasbourg, France, 3Inserm, U977, Strasbourg, France, 4 Pôle de Santé Publique, Centre Hospitalier Régional Universitaire de Strasbourg, Strasbourg, France, 5Pôle Hépato-digestif, Hôpitaux Universitaires de Strasbourg, Strasbourg, France, 6Rega Institute for Medical Research, KULeuven, Leuven, Belgium. *contributed equally Word count: title – 113 characters, abstract – 248 words, text – 4,948 words, 6 figures, 1 table Correspondence: Prof. Thomas F. Baumert, M. D., Inserm Unit 1110, Université de Strasbourg, 3 rue Koeberlé, F-67000 Strasbourg, France; Phone: (++33) 3 68 85 37 03, Fax: (++33) 3 68 85 37 24, e-mail: Thomas.Baumert@unistra . fr Keywords: host-targeting agents; resistance; transplantation; treatment Abbreviations: CI combination index; CLDN claudin; CTRL control; DAA directacting antiviral; HCV hepatitis C virus; HCVcc cell culture-derived HCV; HTA host-targeting agent; IC – inhibitory concentration; IFN interferon ; mAb monoclonal antibody ; MTT 3-(4,5dimethylthiazol-2- yl)- 2,5 - diphenyl tetrazolium bromide; PKI – protein kinase inhibitor. Author contribution I.F. and F.X. contributed equally to this work. T. F. B. initiated, designed and supervised research. I.F., F.X., C.T., R.A., L.M., E.R., P.L., J.N., M.B.Z. and T.F.B. performed research. I.F., F.X., C.T., R.A., L.M., E.R., P.L., J.N., M.B.Z. and T.F.B. analyzed data. N.M. and M.S. performed statistical analyses. F.H. and M.D. provided reagents. I.F., M.B.Z. and T.F.B. wrote the paper. Grant support This work was supported by Inserm, University of Strasbourg, the European Union (ERC-2008AdG-233130-HEPCENT, INTERREG-IV-Rhin Supérieur-FEDER-Hepato-Regio-Net 2009 and 2012, EU FP7 HepaMab), ANRS (2011/132, 2012/239, 2013/ ), Laboratoire d’excellence LabEx HEPSYS (Investissement d’Avenir; ANR-10-LAB-28), the Institut Hopitalo-Universitaire Mix-Surg and the Direction Générale de l'Offre de Soins (A12027MS). The group in Leuven was funded by a grant from the Fund for Scientific Research (FWO) (G.0728.09N) and KULeuven GOA 10/014. Conflict of interest: The authors declare no conflict of interest. Inserm and the University of Strasbourg have filed patent applications on the synergy of interferons, direct-acting antivirals and entry inhibitors as well as SR-BI-specific antibodies, erlotinib, dasatinib for prevention and treatment of HCV infection. Inserm, University of Strasbourg and Genovac have filed a patent application for claudin-1 specific-antibodies for prevention and treatment of HCV infection.	38,668
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La colombe et le serpent, Paris, Classiques Garnier, 2019. VIALA, Alain, Stylistique et sociologie. Classes de postures, Revue belge de philologie et d'histoire, n°93, 2015. VIGNAUX, Georges, Le discours acteur du monde — Énonciation, argumentation et cognition, Paris, Ophrys, « L'Homme dans la langue » [coll.], 1988. WENTZLAFF-EGGEBERT, Christian, Le langage littéraire au début du XVIIe siècle : de la rhétorique à la littérature, Tübingen Günter Narr, 1991. YORKS, Ralph A., « La Rhétorique dans L’Astrée », Dix-septième siècle, n°110-111, 1976, p. 13-24. ZONZA, Christian, « Les discours rapportés dans la nouvelle historique l’exemple de Dom Carlos et du Prince de Condé », dans Histoire, histoires, nouvelles approches de Saint-Simon et des récits des XVIIe XVIIIe siècles, Marc Hersant, Marie-Paule Pilorge, Catherine Ramond et François Raviez [dir.], Arras, Artois Presses Université, 2011, p. 291-306. Ensemble l’Histoire déplorable de la belle Dunglas, vray miroir de constance et chasteté 50, 94 ANONYME [BÉDACIER, Catherine] « Histoire amoureuse du Cardinal de Richelieu », dans Histoire des amour s de Grégoire VII, du Cardinal de Richelieu, de la Princesse de Condé et de la marquise d’ Urfé , 350, 488, 546 ANONYME [BERNARD, Catherine] Le Com te d’Amboise, livre premier et livre second 51, 52, 66, 87, 94, 258, 276 ANONYME [BREMOND, Gabriel ] Hatt igé, ou les amour s du roy de Tamaran , nouvelle 51, 363, 492, 512 ANONYME [CONTI, LouiseMarguerite de Lorraine, princesse de, Mlle de Guise] Les Aventures de la cour de Perse 248, 338 Histoire des amours du Grand Alcandre 87, 149, 533, 534 ANONYME [LA ROCHE-GUILHEM, Anne (de)] Almanzaïde 51 Le Grand Scanderberg, 51, 82, 145, 146, 376 L’Innocente justifiée 139, 141 ANONYME [VAUMORIÈRE, sieur de] Diane de France 51, 52, 475 ANONYME, Isaac] [attribué à CLAUDE, Le Comte de Soissons et le cardinal de Richelieu, rivaux de madame la duchesse d’Elboeuf, 51, 309, 350, 408 ANONYME, [attribué à LA LANDE] Les Adventures non pareilles d’un Marquis Espagnol 136, 137 ANONYME Climandor ou l’histoire des Princes 87, 198, 229, 317 Clorinde, première partie 101, 205, 336, 508 Histoire du comte de Genevois et de Mademoiselle d’Anjou 57, 263 Histoire plaisante et récréative de la belle Marquise fille de Saluste roy de Hongrie 192, 196 La Duchesse de Médo 51, 94, 488 Le Sire d’Aubigny 51, 308 Mémoires du Sérail, sous Amurat second 51, 62, 81, 83, 142, 144, 354, 497, 498 Tachmas, prince de Perse 51, 52, 94, 190, 198, 230, 353, 354, 488 AUBIGNAC, François Hédelin abbé d’ Macarise, ou la Reine des Isles fortunées 73, 87, 90, 92, 153, 154, 163, 238, 269, 286, 290, 369, 466, 467, 469, 470, 471, 486, 487, 535, 536, 537, 567 B BARCLAY, Jean Argenis 10, 20, 32, 33, 34, 42, 86, 90, 222, 223, 224, 235, 275, 286, 316, 349, 370, 371, 372, 373, 375, 376, 378, 429, 430, 432, 471, 536, 560, 561 BAUDOIN, Jean 636 Histoire nègrepontique, contenant la vie et les amours d’Alexandre Castriot 38 BEAULIEU, Sie ur de La Solitude amoureuse du Sieur de Beaulieu 524, 561 BERNARD, Catherine Les Malheurs de l’Amour, nouvelle, Éléonor d’Yvrée 51, 94, 235 première BOISGUILBERT, Pierre le Pesant (de) Mademoiselle de Jarnac 51, 52, 519, 540, 545 BOITEL DE GAUBERTIN, Pierre Histoire tragique de Circé 125, 159 La De faicte du Faux amour 159, 160, 161, 291, 292, 349, 515 BOURSAULT, Édme Artemise et Poliante 169 Le Prince de Condé 51, 52, 53 C CAMUS, Jean-Pierre Aristandre, histoire germanique 106, 275 Alexis 93, 130, 452, 531 Iphigène, 42, 43, 64, 67, 68, 246, 300, 317, 331, 349, 358, 364, 499, 526, 537, 539, 543, 555, 556, 557, 558, 568 Marianne ou l’innocente victime 332 Pétronille, accidents pitoyables de nos jours, cause d’une vocation religieuse 499, 512, 558 CAUMONT LA FORCE, CharlotteRose Mémoire historique ou Ane cdote galante et secrete de la duchesse de Bar, sœur d’Henry IV roy de France. URFÉ, Honoré (d’), L’Astrée, Deuxième partie (1610), Delphine Denis [éd.], Paris, Honoré Champion, 2016. DES ESCUTEAUX, Nicolas, Les Traversez hasards de Clidion et Armirie, par le Sr D. E., Paris, François Huby, 1612. DES ESCUTEAUX, Nicolas, Les Admirables faits d’armes d’Alcestes servant l’infidele Lydie. Par le sieur des Escuteaux, Saumur, Thomas Portau, 1613. ANONYME, Histoire plaisante et récréative de la belle Marquise fille de Saluste roy de Hongrie. En laquelle on pourra voir la constance d’une dame vertueuse, en ses infortunes, et comme Dieu en fin guerdonne ceux qui mettent leur espoir en lui, Lyon, François Arnoullet le vieux, 1615. BOITEL DE GAUBERTIN, Pierre, Histoire tragique de Circé, ou suite de la défaite du faux amour, ensemble l’heureuse alliance du cavalier victorieux et de la belle Adrastre, Paris, Pierre Chevalier, 1617. BOITEL DE GAUBERTIN, Pierre, La Defaicte du Faux amour par l’unique des Braves de ce temps, député à l'exécution d'un acte tant héroïque durant l'absence de Mars, Hercule, Mercure, Apollon et Nestor, dieux fugitifs du ciel, fiction intelligible aux jugemens solides. Dédié au cavalier victorieux, Paris, Pierre Chevalier, 1617. MATTHIEU, Pierre, Ælius Sejanus. Histoire romaine, recueillie de divers autheurs, Paris, Robert Estienne, 1617. MATTHIEU, Pierre, Histoire des prosperitez malheureuses d’une femme Cathenoise, grande Seneschalle de Naples, Paris, Jean Regnoul, 1617. DES ESCUTEAUX, Nicolas, Le Ravissement de Clarinde, Histoire tres-véritable, par le Sieur des Escuteaus, Poitiers, Antoine Mesnier, 1618. GOMBERVILLE, Marin Le Roy ( ), L’Exil de Polexandre et d’Ériclée, Paris, Toussaint Du Bray, 1619. URFÉ, Honoré (d’), L’Astrée, Troisième partie (1619), Hugues Vaganay [éd.], Lyon, Masson, 1927. ANONYME [attribué à LA LANDE], Les Adventures non pareilles d’un Marquis Espagnol, Paris, Jean du Hamel, 1620. CAMUS, Jean-Pierre, La Mémoire de Darie, où se voit l’idée d’une dévotieuse vie et d’une religieuse mort, Par Monseigneur l’Evesque de Belley, Paris, Claude Chappelet, 1620. ANONYME (BARAGNES, Rolin), Histoire, vie et mort de Jacques Cinquiesme Roy d’Ecosse. Ensemble l’ Histoire déplorable de la belle Dunglas, vray miroir de constance et chaste té , Paris , Rolin Baragnes , 1621. 644 BARCLAY, Jean , Johannis Barclaii Argenis, Paris , Nicolas Buon, 1621 . La traduction consult ée est id., L ’ Argenis de Jean Barclay, Pierre de Marcassus [trad.], Paris, Nicolas Buon, 1624. GOMBERVILLE, Marin Le Roy (de), La Carithée de M. Le Roy, Sr de Gomberville, contenant sous des temps, des provinces, & des noms supposés, plusieurs rares & véritables histoires de notre temps, Paris, Pierre Billaine, 1621. CAMUS, Jean-Pierre, L’Alexis, De Monseigneur l’Evesque de Belley. Où sous la suitte de divers pélerinages sont déduites plusieurs Histoires tant anciennes que nouvelles, remplies d’enseignements de Pieté, deux parties, Paris, Claude Chappelet, 1622, 6 vol. DOURLENS, Louis, Les Adventures guerrieres et amoureuses de Licide. Histoire veritable de nostre temps, Par le Sieur L. D. D., Paris, Denis de Cay, 1623. LANNEL, Jean (de), Le Romant satyrique, par Jean de Lannel, escuyer Seigneur du Chaintreau et du Chambort, Paris, Toussaint Du Bray, 1624. CAMUS, Jean-Pierre, Aristandre, histoire germanique, Lyon, Jacques Gaudion, 1624. CAMUS , Jean - Pierre, Alcime , relation funeste , o ù se découvre la main de Dieu sur les impies, Paris , Martin Las nier, 1625. CAMUS Jean-Pierre, L’Iphigène, rigueur sarmatique, Lyon, Antoine Chard, 1625, 2 vol. LANNEL, Jean (de), Le Romant des Indes, par Jean de Lannel, escuyer seigneur du Chaintreau & du Chambort, Paris, Toussaint Du Bray, 1625. CAMUS, Jean-Pierre, Pétronille, accidents pitoyables de nos jours, cause d’une vocation religieuse, Lyon, Jacques Gaudion, 1626. URFÉ, Honoré (d’), BARO, Balthazar, L’Astrée, quatrième partie (1627), Hugues Vaganay [éd.], Lyon, Masson, 1927.	23,778
W2556561652.txt_1	Open-Science-Pile	Open Science	Various open science	2,016	PRIME Partnerships in International Medical Education - Restoring a Christian ethos to medical education worldwide	None	English	Spoken	3,170	4,991	CAPACITY BUILDING PRIME Partnerships in International Medical Education Restoring a Christian ethos to medical education worldwide Huw Morgana a Retired GP/Medical missionary, Medical Educator, Staff Tutor and Executive member of PRIME, Emeritus Director of GP training, Bristol, UK Abstract Modern medicine has developed from an essentially Christian world-view and in Western countries has been greatly influenced by the Christian tradition of hospitality and caring for the sick. However, during the 20th century, medical education became increasingly secularised and focussed on the bio-physical model of disease, losing sight of a holistic view of the person that includes awareness of a spiritual dimension. Former Communist countries in particular have little recent tradition of caring, and medical education there tends to be characterised by poor role-models and out-dated didactic teaching. In the resource poor countries of the global South there are many Christian hospitals and clinics but often a lack of experienced medical teachers. Partnerships in International Medical Education (PRIME)’s vision and mission is to support health-care education worldwide to restore a Christian-based holistic approach to patients, and act as a resource where needed, tailoring medical educational programmes to meet the needs of overseas partners (or colleagues in the NHS). Using interactive leaner-centred and problembased educational methods, PRIME tutors (all experienced and qualified Christian medical educators) seek to model patient-centred care by using learner-centred teaching, valuing each person as a bearer of the image of God. Most of PRIME’s teaching involves the doctor-patient relationship, communication skills, compassion, ethics and professionalism, often based around particular clinical scenarios to suit the learners. Small teams of voluntary tutors visiting partner institutions and colleagues for a few weeks a year can have a surprisingly large impact, as those grasping the vision become advocates for positive change in their own situations. Training of trainers and teachers in learner-centred, androgogic methodology to build capacity and sustainability is also a major part of the work. Introduction PRIME, http://www.prime-international.org is a Christian medical education charity, an international network of Christian healthcare professionals who are involved in teaching their discipline. In this paper, I will briefly outline the historical context of PRIME’s work and the challenges and opportunities we face in contemporary medical education around the world and then describe how PRIME is working to meet them. (“Medical” here is used to include nursing and professions allied to medicine.) Nov 2016. Christian Journal for Global Health, 3(2): 134-139. 135 Historical Context Modern medicine and medical teaching have developed in the Western world, influenced by both the Christian tradition of hospitality pioneered by the European monasteries1, and the Judao-Christian value system that has historically underpinned the major institutions of our society (Government, Law, Education, Commerce, etc.).2 This is particularly the case in Western and Northern European countries that experienced the Protestant Reformation in the 16th century. In addition, it can be argued that the scientific advances that have led to the development and success of modern medicine stem from the Reformation’s effect on thought that saw the investigation of the material world as a legitimate and necessary enterprise in understanding God’s creation (“Thinking God’s thoughts after Him” – Johannes Kepler, 1599).3 The remarkable advances in medicine in the th 19 and 20th centuries such as greatly improved public health, the development of surgery and anaesthesia, immunisation, antibiotics, treatment for hormone deficiencies (insulin, thyroxine), effective treatment for many chronic diseases and for mental illnesses all flowed from this, and Christian doctors were involved in many of these advances. In addition, the missionary movement that started in the 19th century resulted in mission hospitals and community health programmes being established in many remote areas in the majority of the world, bringing the advantages of modern medicine delivered with a Christian ethos to the poor and marginalized in many developing countries.4 The Challenge We Face However, the Reformation also paved the way for the Enlightenment, which gradually eroded belief in God from the scientific and public arenas through the 17th century and following, so that the moral and spiritual framework that had guided the development of modern science and medicine was lost.5 The subsequent atheistic and reductionist approach presupposed that the material world was all that existed and that everything could be understood in Morgan terms of its molecular structure. Thus Francis Crick, one of the discoverers of the structure of DNA, famously said, “You, your joys, sorrows, memories and ambitions are in fact no more than the behaviour of a vast assembly of nerve cells and their associated molecules.” 6 This perspective has tended to dehumanise the practice and teaching of medicine, leading to the loss of understanding that patients, students, and doctors are all human beings made in God’s image. There has been too much emphasis on narrow scientific knowledge, abusive teaching methodologies inappropriate for a caring profession, and lack of a whole person perspective and compassion in teaching and practice.7 So the realities of medical practice in the world today include the following:  Gross inequalities in access to health care within and between nations  Poor governmental management of health care systems  Loss of humanity and compassion by health care workers  Failure to respect human life from conception to natural death  Medical practice increasingly divorced from an ethical framework  The threats of unregulated germ-line gene therapy and selective embryo screening  Lack of acknowledgment of a spiritual perspective in human suffering  Excessive emphasis on science and systems in the teaching and practice of medicine  Gradual loss of idealism and growth of cynicism amongst students and young doctors and nurses 8 In the former communist countries, there tends to be little recent tradition or history of a caring approach to patient-care. There are also frequently only limited, out-date didactic teaching and text books with little exposure to real patients or opportunities for students to develop problem solving skills. There are also often poor rolemodels and widespread corruption in health care and its teaching, where bribes are necessary to be seen or taught by a doctor and to pass examinations.9 In the resource poor countries of Nov 2016. Christian Journal for Global Health, 3(2): 134-139. 136 the global South (such as in Africa, Asia, and South America) the too few doctors and healthcare professionals often have to work in isolation with little opportunity for continuing professional development and so lack appropriate training and skills. There is an understandable tendency for doctors and nurses to immigrate to richer countries to work, further adding to the resource problem.10 A Door of Opportunity There is light dawning, however! The “Modern” era is ending and “Post-Modernism” is challenging the closed scientific world-view of Modernity. People are dissatisfied with dehumanising medical treatments (successful as they may be) and are searching for alternatives.11 Spirituality and concern for the whole person are making a comeback. Many medical schools are teaching communication skills and using humanities to aid reflection on the human condition. In our now electronically interconnected world, there is more awareness of the global inequalities in health-care and a desire to redress them. The World Health Organisation (WHO) reports: Until recently the health professions have largely followed a medical model, which seeks to treat patients by focusing on medicines and surgery, and gives less importance to beliefs and to faith in healing, in the physician and in the doctorpatient relationship. This reductionist or mechanistic view of patients is no longer satisfactory. Patients and physicians have begun to realise the value of elements such as faith, hope, and compassion in the healing process.12 So as this new millennium began, Christians had a great opportunity to reclaim the teaching of Medicine as a spiritual and ethical practice. We need to practice and teach in a way that demonstrates respect for our patients and learners as people made in God’s image. We must also ensure our teaching embodies the concept of Whole Person Care for our patients, including a spiritual perspective, alongside evidence-based Morgan scientific truth. It is in that context that PRIME has developed, and grown remarkably, from small beginnings in the UK twenty years ago to a world-wide network of several hundred Christian medical educators sharing the same vision and mission today. PRIME’s Mission PRIME is a global network of Christian healthcare professionals (Nurses, Doctors, Paramedics) involved in teaching their discipline. There are four major components of our mission:  To support and improve medical education in resource-poor countries  To demonstrate and teach compassionate person-centred medical practice  To model the example of Jesus as the perfect physician and teacher  To restore Christ’s values to medical education and practice All this is with the aim of restoring a Christian ethos to medical education worldwide and so improving health-care for all. It has wisely been said that, “A (medical) teacher. . . can influence the lives of many times more patients through his or her students. . . than through a career of surgeries or clinics.”13 PRIME is linked to the Christian Medical Fellowship (CMF), UK and the International Christian Medical and Dental Association (ICMDA). It responds to specific invitations for partnership from groups of doctors or other health professionals in developing and restructuring countries. It has also worked with groups of pastors in some countries, teaching them basic health care and encouraging them to see this as part of their church’s ministry to the community. Its usual pattern of work is to make recurrent short-term (usually 1-3 week) visits to host countries with teams of 2 or 3 tutors to run teaching programmes and maintain relationships with hosts via email and Skype between visits. It provides teaching and training in person-centred and evidence-based medical practice for underresourced doctors from a Christian perspective and where possible, advises on local curriculum development, training methods, and health care Nov 2016. Christian Journal for Global Health, 3(2): 134-139. 137 Morgan management. PRIME’s teaching is in the context of forming genuine relationships with our hosts (generally essential to work effectively in nonWestern cultures). It emphasizes the importance of a compassionate, whole person approach to patients, builds ethical frameworks for decision making, and includes a spiritual perspective on patient care. Its teaching is learner centred, interactive, and problem based (a patient-centred approach to medical practice needs to be taught using a learner-centred approach).14 It aims for sustainability by training medical teachers in good educational methodology whenever possible. PRIME operates its courses with the approval of the University of Brighton and Sussex Department of Postgraduate Medicine and (where possible) host country university departments. It is staffed by volunteer Christian doctors and nurses with experience and qualifications in medical education, and runs a small office with a part-time manager and administrators. It produces educational materials to spread its vision (mostly electronic on CD’s or online, but with some paper publications). It now has branches in Australia, Portugal, Kenya, and Nigeria that plan and run their own teaching programmes internationally and has specialised Mental Health and Palliative Care sections. Most of PRIME’s teaching focuses on compassionate, holistic care, communication skills, professionalism and ethics sometimes structured around clinical topics. For example, there is a “Values Added” modular course aimed at junior doctors in the early years of their training (based on small groups looking at a DVD/CD and then discussing the questions raised with a more experienced facilitator/tutor) that aims to add a Christian perspective to their standard training and encouragement in its practical application to patient care and teaching. PRIME has found that because all people are made in God’s image, most respond positively to compassion and Godly values displayed in healthcare and its teaching. It has worked with Hindus, Muslims, Buddhists, Sikhs, and Atheists, but all agree about the core principles of good medical practice, and all can see that Jesus was an exemplary physician and teacher in his dealings with people. This enables PRIME to maintain and promote its Christian basis and values whilst being acceptable to people of all faiths and none. Participants are always asked to evaluate PRIME courses and sometimes follow-up studies are conducted a few months later to see if those who took part have sustained any change in practice as a result. Whilst it is very hard to measure this objectively, there have been numerous personal testimonies to the transforming nature of PRIME programmes such as, “Today I became a doctor,” ”Before this I wanted to leave (my country) to earn more money, now I want to stay and serve our people compassionately,” “I now understand that all patients are human beings like me and will treat them accordingly,” and “Now I know how to serve Jesus through practising medicine.” An example – teaching whole person care in Kenya tutors were invited back in 2003 to facilitate the second annual conference (one of the original team and another tutor with previous experience of working in Kenya went on this occasion). The programme for this visit focussed on chronic disease management and good practice organisation to provide holistic care and took place in two centres. Email contact with key Kenyan Christian medical leaders was maintained through the years (one ex-patriot Family Physician in particular), and in 2010, three other PRIME tutors ran a programme at the invitation of KAFP at a conference on empowering medical educational leadership. Then in 2014, one of the original PRIME In 2002, three PRIME tutors (British GP educators, two of whom had worked in Kenya years previously) were invited to Kenya to facilitate the inaugural four-day conference of the Kenyan Association of Family Physicians (KAFP). Many of the members of this were Christians and also members of the Kenyan Christian Medical and Dental Association (CMDA). The programme was a values-based introduction to holistic care, good communication skills, professionalism, and various other topics; all taught interactively with an implicit (and sometimes explicit) Christian perspective. The Nov 2016. Christian Journal for Global Health, 3(2): 134-139. 138 Morgan team to visit Kenya was invited by the ex-patriot family physician in partnership with the leaders of the CMDA to formally launch PRIME in Kenya. The idea was to establish an indigenous PRIME Kenya, to spread the vision of faith-based, person-centred, holistic care throughout the country and wider to other East African countries through members of the CMDA teaching PRIME concepts and using PRIME materials. The core PRIME seminars on “Whole Person Care” and “Teaching to Change Hearts” were taught in a number of centres around the country, and in each place, key local doctors were identified who would be active in taking the vision forward. A Kenyan surgeon based in Nairobi became a very active and enthusiastic advocate and teacher of the PRIME vision around Kenya and in other East African countries, and other doctors with faculty positions in a new medical school were able to incorporate aspects of the PRIME material into the curriculum they were teaching. Thus over a twelve-year period, recurrent visits and maintained personal relationships between PRIME tutors and in-country colleagues led to the forming of a true partnership, with Kenyan partners eventually totally owning and running with the vision themselves in their own situations and more widely across East Africa. To give an idea of PRIME’s activity, in 2015, 115 international PRIME tutors carried out 76 separate programmes in 24 different countries in Africa, Asia, and Europe, providing 687 days of teaching (out of 1451 days in the host country) and training to nearly 4,500 individuals. At current standard NGO consultancy rates for teaching and subsistence, this would have cost £535,000. The actual cost (met by tutors) was £31,850. The fact that tutors give their time and expertise freely, adds powerfully to the Christian impact of the work. There is also the vast amount of work done in-country by national teachers and tutors who replicate the teaching and methods in their day-to-day work and teaching. PRIME believes that this training will improve the care provided to the thousands of patients each of those individuals will treat, or the hundreds of healthcare students they will teach in the course of their careers. So in summary, PRIME teaches and encourages healthcare professionals worldwide to teach and practice whole person medicine, using patient and learner-centred teaching methods. It promotes compassionate medicine using Jesus as a role model and seeks to use the basis of Christian values as a central platform for change wherever it’s tutors work. Any health-care professional committed to a compassionate, holistic approach to patient care is welcome to join the PRIME Network, and those involved in teaching who are able to give a week or two a year at their own expense are welcome to apply to be PRIME tutors (details and application forms from admin@prime-interational.org.uk). In the next few months, PRIME is running training programmes in Australia, Kenya, Nigeria, Peru, and UK. References 1. Crislip AT. From monastery to hospital: Christian monasticism & the transformation of health care in late antiquity. Ann Arbor: University of Michigan Press; 2005. http://dx.doi.org/10.3998/mpub.93465 2. Pannenberg W. Christianity and the West: ambiguous past, Uncertain Future. First Things: Mon J Relig Publ Life. 1994;18. Available from: https://www.firstthings.com/article/1994/12/christi anity-and-the-westambiguous-past-uncertain-future 3. Hookyaas R. Religion and the rise of modern science. Edinburgh: Scottish Academic Press; 1972. 4. Hardiman D, editor. Healing bodies, saving souls: medical missions in Asia and Africa, Wellcome Series in the History of Medicine, Clio Medica 80. Amsterdam and New York: Rodopi; 2006. 5. Israel JI. Radical enlightenment: philosophy and the making of modernity 1650-1750. Oxford: Oxford University Press; 2001. 6. Crick F. The astonishing hypothesis: the scientific search for the soul. London: Simon and Schuster; 1994. 7. Coulehan J. Viewpoint: today's professionalism: engaging the mind but not the heart. Acad Med. 2005;80: 892-8. Nov 2016. Christian Journal for Global Health, 3(2): 134-139. 139 8. 9. 10. 11. Morgan http://dx.doi.org/10.1097/00001888-20051000000004 Steinbeck B, London AJ, Aras JD. Ethical issues in modern medicine: contemporary readings in bioethics. Boston: McGraw-Hill; 2009. Rechel B, McKee M. Health reform in central and Eastern Europe and the former Soviet Union. Lancet. 2009;374(9696):1186-95. http://dx.doi.org/10.1016/S0140-6736(09)61334-9 Chudi IP. Healthcare problems in developing countries. Med Practice Rev. 2010;1(1):9-11. Rayner L, Easthope G. Postmodern consumption and alternative medications. J Sociol. 2011 ; 37 (2) :157-76. Http://dx.doi.org/10.1177/144078301128756274 12. World Health Organization. WHOQOL and spirituality, religiousness and personal beliefs: report on WHO consultation. Geneva: WHO; 1998. Available from: http://apps.who.int/iris/bitstream/10665/70897/1/W HO_MSA_MHP_98.2_eng.pdf 13. Smith BH, Edwards E, Murchie P. Society for academic primary care: scotching the myths. Br J Gen Prac. 2005;55:513 (316). 14. Stewart M. Patient-centered medicine: transforming the clinical method. Oxford: Radcliffe Publishing; 2003. Peer Reviewed Competing Interests: Dr. Morgan is a Staff Tutor and executive member of PRIME Correspondence: Huw Morgan, United Kingdom. jhcmorgan@gmail.com Cite this article as: Morgan H. PRIME Partnerships in International Medical Education Restoring a Christian ethos to medical education worldwide. Christian Journal for Global Health (Nov 2016), 3(2):134-139. © Morgan H. 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 properly cited. To view a copy of the license, visit http://creativecommons.org/licenses/by/4.0/ www.cjgh.org Nov 2016. Christian Journal for Global Health, 3(2):134-139.	29,178
https://openalex.org/W4246408805	OpenAlex	Open Science	CC-By	2,020	Mepolizumab	null	English	Spoken	122	268	Qeios · Definition, February 2, 2020 Open Peer Review on Qeios Open Peer Review on Qeios Mepolizumab National Cancer Institute Qeios ID: GGI97C · https://doi.org/10.32388/GGI97C Source National Cancer Institute. Mepolizumab. NCI Thesaurus. Code C157376. A humanized immunoglobulin G1 (IgG1) monoclonal antibody directed against interleukin- 5 (IL-5) with anti-asthmatic and potential immunosuppressive activity. Upon subcutaneous administration, mepolizumab selectively binds to IL-5, preventing it from associating with interleukin-5 receptor subunit alpha (IL5RA) on the surface of eosinophils and their progenitors. IL-5 plays a role in the regulation of eosinophil development from hematopoietic progenitors as well as eosinophil maturation, differentiation, mobilization, activation, and survival. IL-5 also play a role in the pathogenesis of some phenotypes of hypereosinophilic syndrome (HES). Qeios ID: GGI97C · https://doi.org/10.32388/GGI97C 1/1	41,693
https://openalex.org/W4280550355	OpenAlex	Open Science	CC-By	2,022	ATS-STEM Research Methodology and Evaluation Guidelines	María Carmen Fernández Morante	English	Spoken	24,086	42,458	ATS-STEM Research Methodology and Assessment Guidelines ATS-STEM Research Methodology and Assessment Guidelines ATS-STEM Research Methodology and Assessment Guidelines Carmen Fernández-Morante Carmen Fernández de la Iglesia Beatriz Cebreiro López Enrique Latorre Ruiz Francisco Mareque León ATS-STEM Research Methodology and Assessment Guidelines Carmen Fernández-Morante Carmen Fernández de la Iglesia Beatriz Cebreiro López Enrique Latorre Ruiz Francisco Mareque León ATS-STEM Research Methodology and Assessment Guidelines Carmen Fernández-Morante Carmen Fernández de la Iglesia Beatriz Cebreiro López Enrique Latorre Ruiz Francisco Mareque León ATS-STEM Research Methodology and Assessment Guidelines Carmen Fernández-Morante Carmen Fernández de la Iglesia Beatriz Cebreiro López Enrique Latorre Ruiz Francisco Mareque León D5.1: ATS-STEM Research Methodology and Assessment Guidelines Project Title Assessment of Transversal Skills in STEM (ATS-STEM) Reference: 606696-EPP-1-2018-2-IE-EPPKA3-PI Please cite as: Fernández-Morante, C., Fernández de la Iglesia. J.C., Cebreiro López, B., Latorre Ruiz, E., & Mareque-León, F. (2022). ATS-STEM Research Methodology and Assessment Guidelines. Zenodo. https://doi.org/10.5281/zenodo. 6464593 D5.1: ATS-STEM Research Methodology and Assessment Guidelines Project Title Assessment of Transversal Skills in STEM (ATS STEM) Final version Updated 16th November 2020 Assessment of Transversal Skills in STEM (ATS-STEM) Reference: 606696-EPP-1-2018-2-IE-EPPKA3-PI Please cite as: Fernández-Morante, C., Fernández de la Iglesia. J.C., Cebreiro López, B., Latorre Ruiz, E., & Mareque-León, F. (2022). ATS-STEM Research Methodology and Assessment Guidelines. Zenodo. https://doi.org/10.5281/zenodo. 6464593 Please cite as: Fernández-Morante, C., Fernández de la Iglesia. J.C., Cebreiro López, B., Latorre Ruiz, E., & Mareque-León, F. (2022). ATS-STEM Research Methodology and Assessment Guidelines. Zenodo. https://doi.org/10.5281/zenodo. 6464593 This methodological research design has been developed within the framework of work package 5 of the European project "Assessment of Transversal Skills in STEM (ATS-STEM) and has been carried out by the educational technology group of the University of Santiago de Compostela (http://tecnoeduc.es) and are available from the project website: https://www.atsstem.eu/resources/ Final version Updated 16th November 2020 Final version Updated 16th November 2020 1.1. INTRODUCTION TO THE CONCEPT OF EVALUATIVE RESEARCH 1.1. INTRODUCTION TO THE CONCEPT OF EVALUATIVE RESEARCH The concept of evaluative research is presented as a strategic instrument for decision-making to develop and improve society and the quality of life of citizens. This instrument may be applied in diverse fields such as education, health, economy, culture, social protection and public policies... stressing its transdisciplinary nature, the rise in the assessment of organizations and institutions, its support in various methodologies and the importance of participatory strategies. It highlights the usefulness and appropriate use of assessments as a priority objective of this type of research, always based on ethical and scientific quality principles and standards and the corresponding meta- assessment studies (Escudero, 2016). Evaluative research should therefore be understood as a rigorous, controlled and systematic process of collecting and analysing valid and reliable information to make decisions about an educational program; and thus increase the rationality of decisions about its implementation, development and assessment (Tejedor, García-Varcárcel & Rodríguez, 1994). In this vein, Correa (2002) specifies evaluative research as "a special type of applied research whose goal, unlike basic research, is not the discovery of knowledge. With the main emphasis on usefulness, evaluative research should provide information for program planning, implementation and development. Evaluative research also assumes the particular characteristics of applied research, which allows predictions to become a research result. In relation to general educational research, some fundamental aspects of evaluative research stand out (Tejedor, García-Varcárcel, and Rodríguez, 1992) While the purpose of research is to increase knowledge in order to draw conclusions that can be generalized to other areas, the process of program assessment is one of intervention in decision-making on practical problems limited to a reference group. In research, personal or scientific interest delimits the problem on which explicit hypotheses are launched. Through a replicable process, based on a pre-established design and a usually quantitative methodology, the researcher controls and manipulates the variables to obtain data from a randomized sample, and tries to autonomously achieve objective knowledge. However, in evaluative research, there are numerous aspects that can be assigned by external conditioning factors, such as the topic and/or the interest group, preventing randomized sampling. It is a process that is difficult to replicate without explicit hypotheses, through flexible designs in which it is difficult to control the variables, and through diverse methodologies, the researcher tries to assign value criteria to a given program. TABLE OF CONTENTS ................................................................................................................................................................................................. 1 1. INTRODUCTION ................................................................................................................................................................... 4 1.1. INTRODUCTION TO THE CONCEPT OF EVALUATIVE RESEARCH................................................................................... 4 1.2. PARADIGMS AND METHODOLOGY AROUND EVALUATIVE RESEARCH ....................................................................... 5 2. THE METHODOLOGY FOR THE ASSESSMENT OF THE ATS-STEM PROJECT ......................................................................... 6 2.1. RESEARCH AIMS ........................................................................................................................................................... 6 2.2. DESCRIPTION OF THE METHODOLOGY ......................................................................................................................11 2.3. PROFILES, ROLES, TASKS AND RESPONSIBILITIES OF THE AGENTS INVOLVED IN THE RESEARCH ............................13 2.4. TEMPORISATION ........................................................................................................................................................16 2.5. ETHICAL CONSIDERATIONS ........................................................................................................................................18 2.6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEMENTATION. ..............................26 2.6.1. SELF-ADMINISTERED CORE ATS-STEM LEARNING COMPETENCES ONLINE QUESTIONNAIRE ............................51 2.6.2. CLASSROOM OBSERVATION PROTOCOL .............................................................................................................57 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL ..........................................................................................70 2.6.4. ATS-STEM LEARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) ...........85 2.6.5. ARTIFACT ANALYSIS PROTOCOL ..........................................................................................................................86 2.6.6. INTERVIEW PROTOCOL ........................................................................................................................................95 2.7. DATA TRIANGULATION (INCLUDING EXAMPLES).....................................................................................................100 3. DATA ANALYSIS: BASIC SUPPORT FOR THE ANALYSIS OF QUALITATIVE DATA...............................................................101 4. NATIONAL REPORT: STRUCTURE AND FORMAT. ............................................................................................................104 5. REFERENCES ....................................................................................................................................................................108 3. 3. 1.2. PARADIGMS AND METHODOLOGY AROUND EVALUATIVE RESEARCH The term paradigm is understood as the ideological frame of reference or conceptual context that we use to interpret a reality (Bausela, 2003; Medina & Villar, 1995). The paradigm, therefore, is not oriented per se to an incidence on reality, but rather to the interpretation of it under a determined conceptual prism. García Ramos (1992) makes a comparative study of the differential characteristics of the different approaches, from the objectives to the techniques and methodological resources. And Medina and Villar (1995) specify them in three paradigms (qualitative, quantitative and mixed), four models (behaviourists, humanistic and holistic) and three models of educational assessment (behaviourists, humanistic and holistic). The concrete method of assessment, according to Correa (2002), is evaluative research, where the tools of social research are placed at the service of the ideal consisting in making the judgement process more precise and objective. In its research form, assessment establishes clear and specific criteria that guarantee the success of the process, systematically gathers information, evidence and testimonies from a representative sample of the audiences that make up the program or object to be evaluated, translates this information into evaluative expressions and compares them with the criteria initially established, and finally draws conclusions. The diversity and flexibility in the methodology used in evaluative research is a must due to the its transdisciplinary nature and paradigmatic foundation of this type of research, as well as owing to the multiplicity of fields of work, objectives, objects and contexts of analysis with which the evaluator is obliged to work to elaborate his or her various diagnoses and proposals for action for the different people involved, responsible for, and affected by the research (Christie & Fleischer, 2010; Expósito, Olmedo & Fernández-Cano, 2004). This pluri-reality makes the vast majority of research methodologies and procedures useful for evaluative research. Nevertheless, logically, their applicability is conditioned and defined in each case by the type of problem, aspect or theme being researched (Betzner, Lawrenz, & Thao, 2016; Makrakis & Kostoulas-Makrakis, 2016; Perassi, 2009; White, 2013). The same applies to the procedures and sources for collecting information, a key aspect in many assessment studies, which must be supported by variables, information, opinions, and so forth, which are very diverse in terms of their typology, the diversity of those involved and the possibilities for collection. 1.1. INTRODUCTION TO THE CONCEPT OF EVALUATIVE RESEARCH Generally speaking, for Expósito (2003), evaluative research is similar to any other research process, distinguished more by the objectives than by the methods used. It could therefore be understood as a mere extension, with its own set of characteristics, towards a concrete practical field. The author highlights certain characteristics of evaluative research: • Throughout the process of evaluative research, from the selection of the problem to the application of a specific methodology, it is accompanied by value judgments about the program. The complexity of their object of knowledge and the contexts in which they are carried out make both the formulation of precise hypotheses and a complete control of the intervening variables extremely complex, limiting the use of designs with a more experimental character. • Throughout the process of evaluative research, from the selection of the problem to the application of a specific methodology, it is accompanied by value judgments about the program. The complexity of their object of knowledge and the contexts in which they are carried out make both the formulation of precise hypotheses and a complete control of the intervening variables extremely complex, limiting the use of designs with a more experimental character. • The collection of data is conditioned by the viability of the process and the possibilities of the subjects involved. • The collection of data is conditioned by the viability of the process and the possibilities of the subjects involved. 4. 4. lication, given the peculiar and unrepeatable characteristics of the execution, is practically impossible. • Replication, given the peculiar and unrepeatable characteristics of the execution, is practically imposs • Decisions on stopping, replacing or repeating the program are not usually left to the evaluator; the report is adapted to the requirements of the financier. • In other aspects, while research finds its validity internally, externally and through theoretical constructs, data are interpreted according to defined rules. Reports are adapted to the scientific community; program assessment, bases its credibility on the very usefulness that allows the assessment of the program, adapting its interpretations to managers and users. • In other aspects, while research finds its validity internally, externally and through theoretical constructs, data are interpreted according to defined rules. Reports are adapted to the scientific community; program assessment, bases its credibility on the very usefulness that allows the assessment of the program, adapting its interpretations to managers and users. 1.2. PARADIGMS AND METHODOLOGY AROUND EVALUATIVE RESEARCH This complexity that the evaluator usually faces, makes the triangulation of methods, techniques, sources and procedures of information collection and the combined use of different methodologies (mixing methods) acquire a special relevance in the evaluative research, as a means of strengthening and validating the diagnoses and results. 5. 5. 2. THE METHODOLOGY FOR THE ASSESSMENT OF THE ATS-STEM PROJECT 2.1. RESEARCH AIMS On the previous advances of the ATS-STEM consortium and the conceptualization of Integrated STEM Education proposed under WP1 and WP2, we have designed an evaluative research that will allow us to collect evidence throughout the Pilot process, combining both quantitative and qualitative techniques. This research will involve a rigorous, controlled and systematic process of collecting and analysing reliable and valid information to make decisions on the ATS-STEM programme and to provide recommendations on the implementation of integratedATS-STEM projects in European schools. Thus, this research methodology aims to generate useful ways of understanding educational innovation, and use it as a critical resource to improve educational action processes (Tejedor et al., 1994; Pérez Juste, 1995, Amezcua and Jiménez, 1996, Martínez Mediano, 1997). The objective of the research developed will therefore be two-fold. On the one hand, (I) "knowing and explaining" the process of implementation of the ATS-STEM programme, and on the other, (II) "understanding" how it works in different contexts (schools, classrooms or countries) hence, making proposals to improve it. In this respect, it is very important, as we will see, that the methodological proposal is perfectly integrated into the design of the ATS-STEM educational proposal and its adaptation to the context of the different schools participating in the implementation process. Through this research, we hope to obtain evidence that will allow us to analyse the possibilities of digital assessment in the implementation of the teaching of STEM skills and competences in European schools. Thus, the research will gather evidence on the use of digital technology as a resource to improve the way we assess student learning inATS-STEM projects. Throughout the process, the research questions will be very much in the forefront, focusing our attention on these two specific areas, digital assessment and STEM education. However, we believe it is crucial to emphasize that the research objective is not to evaluate a particular set of tools, but rather the populations of digital assessment as a whole in the application of STEM methodologies. We therefore ask ourselves: • How could digital assessment practices support the development of STEM skills? • How could digital assessment practices support the development of STEM skills? • What are the challenges of using digital assessment strategies in STEM learning? • Why apply digital assessment in the development ofATS-STEM projects? • What and how does it contribute to STEM teaching and learning processes? 2. THE METHODOLOGY FOR THE ASSESSMENT OF THE ATS-STEM PROJECT • How and with which digital assessment methodologies can we improve STEM teaching and learni • How and with which digital assessment methodologies can we improve STEM teaching and learning processes? In order to answer these research questions and to obtain the necessary data that will allow us to formulate a suitable analysis of our results, the research will be developed by combining not only different techniques (quantitative and qualitative), but also different instruments. In order to answer these research questions and to obtain the necessary data that will allow us to formulate a suitable analysis of our results, the research will be developed by combining not only different techniques (quantitative and qualitative), but also different instruments. • Two-phase self-administered electronic questionnaire on students' perception of the development of the eight STEM competencies identified in the project. • Two-phase self-administered electronic questionnaire on students' perception of the development of the eight STEM competencies identified in the project. • Observation sheet of the digital assessment tools for the analysis of the interaction, communication, collaboration and reflection dynamics generated by the digital assessment instruments implemented in eachATS- STEM project. These data will allow us to understand the role and possibilities of digital assessment in the implementation of the ATS-STEM methodology. • Observation sheet of the digital assessment tools for the analysis of the interaction, communication, collaboration and reflection dynamics generated by the digital assessment instruments implemented in eachATS- STEM project. These data will allow us to understand the role and possibilities of digital assessment in the implementation of the ATS-STEM methodology. 6. 6. • Classroom observation sheet with which to record the data obtained through the observation of some sessions throughout the pilot and which will provide objective information to help understand the process of application of the ATS-STEM methodology, the dynamics of interaction and the processes of knowledge production generated, the functions adopted by teachers and students and the difficulties encountered in the application of the programme. • Artifact analysis sheet with which to analyse students' creations such as reflections, video creation, podcasts, multimedia and hypermedia materials, computer graphics... and teachers' feedback on these tasks. • Artifact analysis sheet with which to analyse students' creations such as reflections, video creation, podcasts, multimedia and hypermedia materials, computer graphics... and teachers' feedback on these tasks. 2. THE METHODOLOGY FOR THE ASSESSMENT OF THE ATS-STEM PROJECT • Interviews with teachers, students and mentors from the ATS-STEM programme participating in the pilot to reflect on the process, learn about students' perceptions, teachers' perceptions of digital assessment and STEM education and see if these evolved over the course of the project. This will allow us to carry out a meta-analysis of the teaching-learning process. This procedure will allow us to carry out a triangulation of the types of data, instruments and informants that will guarantee the validity of the investigation and help us to formulate hypotheses and endorse the explanations that we develop. Data triangulation: Triangulation of methods Source triangulation - informants Strategy and description of data collection instruments The aim of our evaluative research is to analyse the possibilities of digital assessment in the implementation of teaching STEM skills and competences in European schools. The research will gather evidence on the use of digital technology to improve the way we assess student learning. It will seek to determine the strengths and impacts on the implementation of STEM methodologies by answering questions such as - How might digital assessment practices support the development of STEM competencies? How might digital assessment practices support the development of STEM competencies? - What are the challenges of using digital assessment strategies in STEM learning? - Why apply digital assessment in the development of ATS-STEM projects? - What and how does it contribute to STEM teaching and learning processes? - How and with which digital assessment methodologies and tools can we improve STEM teaching and learning processes? - How and with which digital assessment methodologies and tools can we improve STEM teaching and learning processes? 7. 7. Description Method and priorities ROLES Dates onic questionnaire for nts with which to measure rception of success in the ition of STEM skills. nts participating in the t through the 20 pilot s will complete the onnaire before starting their pation in the project and at d dents participating in ATS- projects (pilot schools and tudy) Quantitative analysis. For the construction of the questionnaire, the main theoretical and bibliographical references that support the research on the competences identified in wp1 as core stem competencies have been taken into account Different questionnaires prepared for other investigations that were directly or indirectly related to the core ATS-STEM learning competences have also been reviewed. USC produces the instrument. Pilot partners translate it into their national languages. The USC implements it electronically and analize all the international data. PRODUCT : Electronic Questionnaire for Students in all languages (DOC1) May 15th 31st May It will depend on the pilot's final dates As a guideline, December-January 2021 and June 2021 oom observation record to documenting what ned in the classroom ing) sessions held during the about the formative ment process and the mentation ofATS-STEM ts studies from among the 20 chools per country ervations per learning cycle conducted in a minimum of ning cycles per school. Qualitative Analysis This record sheet will provide evidence for the analysis of the dynamics of interaction, communication, collaboration and reflection generated from the formative assessment strategy. Strategy and description of data collection instruments For the construction of this record sheet, we have focused our attention on identifying categories of analysis from what the wp2 partners have called Key esoteric of formative assessment tasks. USC produces the instrument. The national researcher of each country is in charge of collecting the data. The national researcher of each country produces the national report with the qualitative analysis. PRODUCT : First week of June September 2021 Method and priorities For the construction of this record sheet, we have focused our attention on identifying categories of analysis from what the wp2 partners have called Key esoteric of formative assessment tasks. Check List and Record Sheet (DOC2) l assessment tools Record s to document evidence of rmative assessment process he implementation of ATS- projects through the digital sment tools used by teachers. e studies from among the 20 chools per country ervations per learning cycle in mum of 2 learning cycles per l. Qualitative Analysis This record sheet will provide us with evidence for the analysis of the dynamics of interaction, communication, collaboration and reflection generated from the formative assessment strategy and the use of digital assessment tools. These data will allow us to understand the role and possibilities of digital assessment in the implementation of ATS-STEM projects For the construction of this record sheet we have focused our attention on identifying categories of analysis from what the wp2 partners have called Key features of formative assessment tasks. USC produces the instrument. The national researcher of each country is in charge of collecting the data. The national researcher of each country produces the national report with the qualitative analysis. PRODUCT : Check List and Record Sheet (DOC3) First week of June September 2021 act recording and analysis to document the level of etence acquired through ent types of materials ced by the students in the ng tasks proposed by the er. e studies from among the 20 chools per country Qualitative analysis This registration sheet will allow us to obtain data on the course of the learning process through a still photograph such as student productions (texts, reflections, results of activities, videos etc). For the construction of this record sheet we have focused our attention on identifying categories of analysis from what the wp2 peers have called Key features of formative assessment tasks and the wp1 core stem competencies peers. USC produces the instrument. 2.2. DESCRIPTION OF THE METHODOLOGY The research developed is focused on the assessment of the effectiveness of the use of tools and digital support in the implementation of ATS-STEM projects in secondary education in order to facilitate their implementation and therefore, the achievement of core ATS-STEM learning in secondary students. Therefore, the main objective of the assessment is to check how the support of digital tools helps to develop core ATS-STEM learning competences in students. Study design: this is a cross-sectional, multi-center, descriptive observational study whose sample is made up of the students and teams of teachers from the case studies (two per country participating in the pilot study). Study population and sample size: all students in two ATS-STEM project teams from two of the pilot secondary schools in each of the case study countries (Belgium, Cyprus, Ireland, Finland, Slovenia, Spain, Sweden) will be included. In addition, 18 other centers will be chosen for the partial implementation of the research project (pilot centers). In order to carry out this study, the pilot centers and collaborators will request the participation of the minors to their families following the usual procedure of information flow and request of participation of the minors in the activities of their institution. A non- probabilistic sampling will be carried out for the convenience of the participating centers, since it will depend on the inclusion of the institution in the European Project, and, therefore, in the interest of the educational community of the institution to form part of the project. Since the participants are chosen for their proximity and for convenience, we will not a determined number of participants as a starting point. Data analysis: Initially a descriptive analysis of all variables will be performed by calculating frequencies, percentages and association between variables as appropriate. The total score of the ATS-STEM competencies will be established by assigning percentages divided into quartiles to the absolute frequencies of the successful responses. There are two different ways in which a school can participate in the pilot assessment process and thus assist in the implementation of the ATS-STEM project. This distinction responds only to research criteria and implies a different level of commitment and responsibilities during the implementation of the pilot. We refer to the difference between pilot schools and participating schools as a case study. Taking into account this fact, the data to be collected will be of a different nature. Strategy and description of data collection instruments The national researcher of each country is in charge of collecting the data. The national researcher of each country produces the national report with the qualitative analysis. First week of June September 2021 This record sheet will provide us with evidence for the analysis of the dynamics of interaction, communication, collaboration and reflection generated from the formative assessment strategy and the use of digital assessment tools. These data will allow us to understand the role and possibilities of digital assessment in the implementation of ATS-STEM projects For the construction of this record sheet we have focused our attention on identifying categories of analysis from what the wp2 partners have called Key features of formative assessment tasks. This registration sheet will allow us to obtain data on the course of the learning process through a still photograph such as student productions (texts, reflections, results of activities, videos etc). For the construction of this record sheet we have focused our attention on identifying categories of analysis from what the wp2 peers have called Key features of formative assessment tasks and the wp1 core stem competencies peers. 9. A minimum of three per learning cycle of case study with different levels of achievement PRODUCT : registration sheet (DOC4) Interview protocols 2 case studies from among the 10 - 20 pilot schools per country In each of the case studies: - Interviews with teachers - Interviews with students - Interviews with the mentor. Qualitative analysis The interviews with teachers, students and mentors will allow us to know their perceptions of the process, the uses and benefits of the digital assessment and the integrated STEM methodology to produce a meta- analysis of the learning process (As a reflective conversation). USC produces the instrument. The national researcher of each country is in charge of collecting the data. The national researcher of each country produces the national report with the qualitative analysis. PRODUCT : Teachers' Protocol (DOC5) Student Protocol (DOC6) Mentorship Protocol (DOC7) First week of June September 2021 10 2.2. DESCRIPTION OF THE METHODOLOGY Pilot Schools (FIG1) are all those schools that participate in the implementation of the ATS-STEM teaching methodology by putting at least 2 learning cycles into practice. These schools will participate in the quantitative research process by collaborating with an ad hoc questionnaire in two phases to assess the core ATS-STEM learning of the students: at the beginning and at the end of the project. We will have a sample of approximately 120 schools across 7 countries. On the other hand, we refer as case study schools (FIG2) are all those schools that, besides being a pilot school and therefore implementing at least two learning cycles according to the ATS-STEM methodology of integrated projects and participating in the quantitative research, have a greater role in the research since they also participate in the qualitative research. This implies that observations will be made in the classroom, the different productions of the students will be analysed and interviews will be carried out with both the teachers involved and the students. In no case will personal data of the participants be collected, assigning the center a participating institution code and only collecting identifying information of the study as the school year or group. Example: School 001, 3 ESO. Bullet 2. Figure 1. Pilot schools. Figure 2. Schools case study Figure 1. Pilot schools. Figure 1. Pilot schools. Figure 2. Schools case study 12 PROFILES ROLES TASKS AND RESPONSIBILITIES OF THE AGENTS INVOLVED IN THE RESEAR • The National Evaluator (NE): This is the national researcher responsible for collecting and safeguarding the qualitative data following research protocol, monitoring the correct administration of the STEM skills questionnaire, guaranteeing the correct anonymisation of the information and preparing the national research report (FIG 4). 2.3. PROFILES, ROLES, TASKS AND RESPONSIBILITIES OF THE AGENTS INVOLVED IN THE RESEARCH In order to guarantee the correct development of the data collection process and its subsequent analysis, it is convenient to briefly present the different roles, responsibilities and tasks that we will carry out during the research, with special emphasis on the figure of the National Evaluator (from now on NE) responsible for national research. In this sense, we can distinguish up to four different profiles that will participate in the research process, with different levels of commitment and unequal presence in the classroom. Among them, three are informants: namely: Teachers, Mentors and Students. Let us look at them briefly: • Mentors are advisors in the implementation of the ATS-STEM model to the national reality of each territory. In this sense, they can participate with the Partners in the training of teachers in their country, although their fundamental task is to support teachers in the different practical difficulties that may arise during the development of the pilot (FIG 3). • The teaching staff is the team of professionals in charge of the teaching activity during the practical development of the ATS-STEM model. Accordingly, they are responsible for designing theATS-STEM projects and communicating with the NE prior to their implementation by providing them with the design model developed including objectives, activities, timing/duration of each activity and digital assessment procedures (Learning Cycle Template). In addition, it should enable the collection of research data during the pilot case studies, allowing the NE access to both the classroom and the virtual environment (FIG 3). • The students are the agents whose main role in the development of the project is the active participation from the academic field (FIG 3). • The students are the agents whose main role in the development of the project is the active participation from the academic field (FIG 3). 13 13 • The National Evaluator (NE): This is the national researcher responsible for collecting and safeguarding the qualitative data following research protocol, monitoring the correct administration of the STEM skills questionnaire, guaranteeing the correct anonymisation of the information and preparing the national research report (FIG 4). 14 Figure 3. Roles and responsibilities for case study participants Figure 4. Roles and responsibilities of the National Evaluator Figure 3. Roles and responsibilities for case study participants Figure 3. Roles and responsibilities for case study participants Figure 4. Roles and responsibilities of the National Evaluator Figure 4. 2.3. PROFILES, ROLES, TASKS AND RESPONSIBILITIES OF THE AGENTS INVOLVED IN THE RESEARCH Roles and responsibilities of the National Evaluator Figure 4. Roles and responsibilities of the National Evaluator 15 15 2.4. TEMPORISATION With regard to the temporal planning of the implementation of the pilot, the period of data collection and the period of data analysis, the health crisis resulting from COVID-19 has affected the initial planning of the project. Therefore, we have readjusted the times so that the health measures taken do not affect the quality of the research. The implementation period of the pilot will be from October 2020 to June 2021, thus having a duration of 9 months. From that date, the national evaluator will have 2 months to prepare his or her national assessment report following the indications set out in this guide. The deadline for submission of this national report is 31 August 2021. From the reception of all the national research reports, the team of the University of Santiago will elaborate the transnational assessment report (D5.2) and the final executive report (D5.3). Figure 5 shows an outline summarising this process and the deadlines. To make everyone aware of their obligations, this table shows who the national evaluator and the coordinating partner of the project is in each country. To make everyone aware of their obligations, this table shows who the national evaluator and the coordinating partner of the project is in each country. Country Partner coordinator National Evaluator Belgium Bart Van Dyck bart.van.dyck@g-o.be Rudi Hendrickx rudi.hendrickx@g-o.be Cyprus Nicolas Kanaris kanaris.n@cyearn.pi.ac.cy Despo Nicolaidou nicolaidou.de@cyearn.pi.ac.cy Ireland Eamon Costello eamon.costello@dcu.ie Colette Kirwan Finland Jarmo Viteli jarmo.viteli@tuni.fi Iida Maria Peltoma iida-maria@proedugo.fi Slovenia Simona Slavič Kumer simona.slavic-kumer@zrss.si Justina Erčulj Justina.erculj@guest.arnes.si Spain María Luz Ares Fandiño maria.luz.ares.fandino@xunta.gal rebeca villaverde lópez asesoriadocente.tic@edu.xunta.es Sweden Eva Hartell ehartell@kth.se Helena Lennholm lennholm@kth.se 16 16 Figure 5. Timeframe for assessment. Figure 5. Timeframe for assessment. Figure 5. Timeframe for assessment. 17 2.5. ETHICAL CONSIDERATIONS This research study will be carried out in accordance with current legislation. Quantitative data will be obtained from anonymous questionnaires. Since the research team does not know the personal data of the participants, the processing of information collected in the context of this study will be carried out in a dissociated manner, in accordance with the General Data Protection Regulation (Regulation EU 2016-679 of the European Parliament and Council of 27 April 2016) and the Spanish regulations on personal data protection in force. None of the data collected in the context of this research will allow the identification of the participants. Furthermore, the analyses will be carried out by a different researcher than the one physically at the center. Regarding the data collected through observations in the classroom, in virtual environments and interviews, the collection of personal data will be avoided so that participants cannot be identified. Prior to the collection of data, teachers, students and parents or guardians of students will be informed in detail about the study so that they can cover, if they consider it, the consent of voluntary participation in the study in which the anonymity of the participants will be guaranteed. Three models of informed consent have been developed for this purpose, which will only cover the participants of the case study schools: It will be the responsibility of the members to keep a copy of the consents. It will be the responsibility of the members to keep a copy of the consents. • Consent for teachers to participate in the ATS-STEM research project • Consent for one parent or legal guardian to participate in the ATS-STEM research project • Informed consent for the mature child (12-17 years old) to participate in a research study Information about the study will be given to the families of the participating minors in a letter included with the consent form. The participating centers will be responsible for requesting family authorisation to participate in the educational project, following the usual communication procedures. The document of consent of the school of the participating students as well as that of the teachers participating in the project will be delivered to the school and the research team will keep a copy of it. Information about the study will be given to the families of the participating minors in a letter included with the consent form. 2.5. ETHICAL CONSIDERATIONS The participating centers will be responsible for requesting family authorisation to participate in the educational project, following the usual communication procedures. The document of consent of the school of the participating students as well as that of the teachers participating in the project will be delivered to the school and the research team will keep a copy of it. 18 [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] Researcher's name: _ Researcher's name: _ INFORMED CONSENT OF PARTICIPATION IN THE ATS-STEM PROJECT: TEACHER □ YES, I agree to participate in the data collection process of the ATS-STEM project. □ I do NOT agree to participate in the data collection process of the ATS-STEM project. nderstand that my participation is voluntary, and that I may withdraw from the study at any time. ata collected will be analysed and presented without containing any personal data. • I have the right to confidentiality of my data so that any researcher who has access to the information collected will be obliged to keep it confidential. _ Course: _ CONSENT FOR TEACHERS TO PARTICIPATE IN THE ATS-STEM RESEARCH PROJECT TITLE OF THE STUDY: ASSESSMENT OF TRANSVERSAL COMPETENCES IN STEM Dear teacher, We are pleased to inform you that your school is participating in the European Project "Assessment of transversal competences in STEM (ATS-STEM). (REF 606696-EPP-1-2018-2-IE-EPPKA3-PI-POLICY). This project is funded by the European Commission and involves 12 educational institutions from 8 European countries. The objective of the ATS-STEM project is to provide teachers and students with a model of formative assessment supported by tools and virtual learning environments that help students to acquire transversal Core ATS-STEM learning . This model will be implemented and evaluated as part of a pilot project in groups of secondary education students. As part of the research, several data will be collected in your school during the 2020-2021 academic year. Specifically, your participation as a teacher implies that you will: a) provide the national evaluator with the ATS-STEM project designs, b) facilitate access and the assessment agenda since you will be observed during the ATS-STEM projects, c) select and provide productions elaborated by the students during the project and d) manage the informed consent of the families of the participating students. No personal data will be collected during the research. For further information related to the ATS-STEM project you can visit the project website http://www.atsstem.eu/. If you have any questions, please contact Carmen Fernández Morante carmen.morante@usc.es or Carmen Fernández de la Iglesia c.delaiglesia@usc.es Thank you very much for your collaboration. 19 INFORMED CONSENT OF PARTICIPATION IN THE ATS-STEM PROJECT: TEACHER □ YES, I agree to participate in the data collection process of the ATS-STEM project. □ I do NOT agree to participate in the data collection process of the ATS-STEM project. • I understand that my participation is voluntary, and that I may withdraw from the study at any time. • I freely and unselfishly consent to my participation in this project. • The data collected will be analysed and presented without containing any personal data. • I will be able to access the results of this research through the publications made. • I have the right to confidentiality of my data so that any researcher who has access to the information collected will be obliged to keep it confidential. Teacher's name and surname: _____________________________________________ School: ____________________________________________________ Course: ________________ Country: ____________________________________________________________________ Signature Date Researcher's name: ________________________________________________ Signature Date Name and surname of the person reporting: _________________________________________________________________________ Signature Date [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] CONSENT FOR ONE PARENT OR LEGAL GUARDIAN TO PARTICIPATE IN THE ATS-STEM RESEARCH PROJECT TITLE OF THE STUDY: ASSESSMENT OF TRANSVERSAL COMPETENCES IN STEM Dear Parent, Signature Date Name and surname of the person reporting: Signature Date 20 In accordance with the Law on Data Protection and Guarantees of Rights and Regulations EU 2016/679, the data collected will be processed by the University of Santiago de Compostela (USC) for the purpose of "Management of research data", Which aims at managing the data of the European project "Assessment of transversal competences in STEM (ATS-STEM)" (REF 606696-EPP-1-2018-2-IE-EPPKA3-PI-POLICY) to create research results for scientific purposes. The person in charge of data handling is the Vice-Rectory of Research and Innovation, whose contact details are Edificio4 CACTUS, 1ª Planta, 15782 Santiago de Compostela, vr.investigacion@usc.es The Data Protection Delegate is Mr. José Julio Fernández Rodríguez, dpd@usc.es The Data Protection Delegate is Mr. José Julio Fernández Rodríguez, dpd@usc.es The data processing is justified by the consent of the persons concerned. Interested parties can exercise their rights of access, rectification, suppression, limitation of treatment, opposition and portability through the Electronic Headquarters of the USC https://sede.usc.es/sede/publica/catalogo/procedemento/55/ver.htm Interested parties can exercise their rights of access, rectification, suppression, limitation of treatment, opposition and portability through the Electronic Headquarters of the USC https://sede.usc.es/sede/publica/catalogo/procedemento/55/ver.htm Interested parties can exercise their rights of access, rectification, suppression, limitation of treatment, opposition and portability through the Electronic Headquarters of the USC https://sede.usc.es/sede/publica/catalogo/procedemento/55/ver.htm They can also contact the Spanish Data Protection Agency to make any claims they deem appropriate. The data will be kept for a period of time that will not be used to collect them, or for the time necessary to comply with legal requirements. Once the purpose has been fulfilled, the data will be blocked until the applicable deadlines have passed. The privacy and data protection policy of the USC can be consulted on the website http://www.usc.es/gl/normativa/protecciondatos/Politica-privacidade.html If the required data and the authorization for its treatment are not provided, it will not be possible to process it upon request. If the required data and the authorization for its treatment are not provided, it will not be possible to process it upon request. I hereby give my consent to the processing of the data provided in accordance with the applicable legislation. I hereby give my consent to the processing of the data provided in accordance with the applicable legislation. 21 21 Dear Parent, We are pleased to inform you that your child's school is participating in the European Project "Assessment of transversal competences in STEM (ATS-STEM). (REF 606696-EPP-1-2018-2-IE-EPPKA3-PI-POLICY) whose main researchers are J. Carmen Fernández de la Iglesia and Carmen Fernández-Morante. This project is funded by the European Commission and involves 12 educational institutions from 8 European countries. The objective of this project is two-fold: on the one hand, to implement STEM experiences in the classrooms that allow secondary education students to acquire Core ATS-STEM learning (problem solving, innovation and creativity, communication, critical thinking, metacognitive skills, collaboration, self-regulation and disciplinary competences). On the other hand, it aims to provide teachers and students with a model of formative assessment supported by tools and virtual learning environments that help students to acquire transversal Core ATS-STEM learning . As part of the research, various data will be collected at your school. Students will fill in a questionnaire in electronic format before and after the implementation of theATS-STEM project during the 2020-2021 academic year, to check the impact of the project on the acquisition of Core ATS-STEM learning . Furthermore, students will be observed at various times throughout the project, participate in a group discussion focus (recorded on audio) and analyse the work developed during the project that can be used for ATS-STEM proposals. We ask for your consent to create an account for your child for the use of a virtual learning environment and digital tools that allow the implementation of formative assessment methods in the ATS-STEM learning cycles to support the development of Core ATS-STEM learning . For further information related to the ATS-STEM project you can visit the project website http://www.atsstem.eu/. If you have any questions, please contact Carmen Fernández Morante carmen.morante@usc.es or Carmen Fernández de la Iglesia c.delaiglesia@usc.es Thank you very much for your collaboration. We hope to have the opportunity to work with your child using innovative learning methodologies. 22 INFORMED CONSENT STUDENT PARTICIPATION IN THE ATS-STEM PROJECT (2020-2021) with DNI/ID number/passport number _ Course: _ _ Parent/legal guardian of _ _ Parent/legal guardian of _ □ I DO consent to my child's participation in the ATS-STEM project described in the above information. □ I DO consent to my child's participation in the ATS-STEM project described in the above information. □ I DO NOT consent to my child's participation in the ATS-STEM project described in the above information. □ I DO consent to my child's participation in the ATS-STEM project described in the above information. □I DO NOT t t hild' ti i ti i th ATS STEM j t d ib d i th b i f ti □ I DO NOT consent to my child's participation in the ATS-STEM project described in the above information. OT consent to my child's participation in the ATS-STEM project described in the above information. • I understand that my and my child's participation is voluntary and that I may withdraw from the study at any time without reason and without affecting my child's education. • I freely and unselfishly consent to my child's participation in this project. • The data collected from my child will be analyzed and presented in a completely anonymous manne • I will be able to access the results of this research through publications. • I will be able to access the results of this research through publications. • I have the right to the confidentiality of my data so that any researcher who has access to the information collected will be under an obligation of confidentiality. Researcher's name: _ Researcher's name: _ Signature Date Signature Date Name and surname of the person reporting: _ 23 23 Signature Date 24 24 [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] [School logo] [Logo of the institutions] [EUROPEAN PROJECTO LOGO] Signature Date Signature Date In accordance with the Law on Data Protection and Guarantees of Rights and Regulations EU 2016/679, the data collected will be processed by the University of Santiago de Compostela (USC) for the purpose of "Management of research data", Which aims at managing the data of the European project "Assessment of transversal competences in STEM (ATS-STEM)" (REF 606696-EPP-1-2018-2-IE-EPPKA3-PI-POLICY) to create research results for scientific purposes. The person in charge of the treatment is the Vice-Rectory of Research and Innovation, whose contact details are Edificio4 CACTUS, 1ª Planta, 15782 Santiago de Compostela, vr.investigacion@usc.es The Data Protection Delegate is Mr. José Julio Fernández Rodríguez, dpd@usc.es e Data Protection Delegate is Mr. José Julio Fernández Rodríguez, dpd@usc.es The data processing is justified by the consent of the persons concerned. The data processing is justified by the consent of the persons concerned. Except for legal provisions, the data will not be transferred. Except for legal provisions, the data will not be transferred. The interested persons can exercise before the responsible the rights of access, rectification, suppression, limitation of treatment, opposition and portability through the Electronic Headquarters of the USC https://sede.usc.es/sede/publica/catalogo/procedemento/55/ver.htm They can also contact the Spanish Data Protection Agency to make any claims they deem appropriate. They can also contact the Spanish Data Protection Agency to make any claims they deem appropriate. The data will be kept for a period of time that will not be used to collect them, or for the time necessary to comply with legal requirements. Once the purpose has been fulfilled, the data will be blocked until the applicable deadlines have passed. The data will be kept for a period of time that will not be used to collect them, or for the time necessary to comply with legal requirements. Once the purpose has been fulfilled, the data will be blocked until the applicable deadlines have passed. The privacy and data protection policy of the USC can be consulted on the website http://www.usc.es/gl/normativa/protecciondatos/Politica-privacidade.html The privacy and data protection policy of the USC can be consulted on the website http://www.usc.es/gl/normativa/protecciondatos/Politica-privacidade.html If the required data and the authorization for its treatment are not provided, it will not be possible to process it upon request. I hereby give my consent to the processing of the data provided in accordance with the applicable legislation. 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME As we have seen in the methodological framework that we have presented, we will collect data for our research through four qualitative and one quantitative instrument. Following a systematic strategy of triangulation of information and given that our research focuses on how digital formative assessment strategies reinforce the acquisition of Core ATS-STEM learning through the implementation of integrated projects, we have considered as a fundamental basis for the assessment of the pilot, the key characteristics of formative assessment. According to the ATS-STEM framework they are six dimensions: (DOC reff D1.3.) 1. Integrate STEM content (and learning outcomes/goals) 1. Integrate STEM content (and learning outcomes/goals) 2. Reflect STEM learning design principles (and social constructivist views of learning) 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner) 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner) 4. Facilitate peer-assessment (improve learning by activating students as instructional resources for one another) 4. Facilitate peer-assessment (improve learning by activating students as instructional resources for one another) 5. Facilitate self-assessment (improve learning by activating students as owners of their own learning) 5. Facilitate self-assessment (improve learning by activating students as owners of their own learnin 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success) 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success) We have reworked these six dimensions as categories and subcategories of analysis with their consequent indicators, so that they allow us to focus on those aspects of relevance to our research, facilitating the mapping of information and the triangulation of data through cross-checking. All this information is synthesised in the following table which is essential both for planning and guiding the observations as well as for the National Research Report. This table is not a tool in itself, but is the guide that directs the observation throughout the process of data collection and analysis. From this point on, we will refer to it as Categories and subcategories of data analysis. Here we can see in detail what the indicator looks like, who the informant is, what kind of information we collect and through which instrument. CONSENT FOR THE MATURE MINOR (12-17 YEARS OLD) TO PARTICIPATE IN A RESEARCH STUDY STUDY TITLE: Assessment of transversal competences in STEM RESEARCH COORDINATOR: ________________________ SCHOOL ____________________ • have read the information sheet given to me by the above-mentioned study participant and was able to talk to:_________________________________________________ and ask any questions about the study. • understand that my participation is voluntary, and that I may withdraw from the study at any time, without reason and without any impact on my medical care. • agree that my data may be used under the conditions detailed in the participant information sheet. • freely agree to participate in this study. At the end of this study I accept that my data are: Deleted Preserved anonymized for future use in other research _______________________________________________ _____________ _____________ Participant's name and surname SIGNATURE Date _______________________________________________ _____________ _____________ Name of legal representative/parent SIGNATURE Date (One of the two parents can sign if the other parent is known to be unopposed) _______________________________________________ _____________ _____________ Name and surname of the researcher SIGNATURE Date Participant code assigned after signing this informed consent: M-______________________________ 25 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME It is also important to keep in mind that we need concrete examples of classroom situations in order to prepare national and transnational reports. All the instruments developed from these categories, also include a section of contextualization and common metadata, which identify the school (educational level, number of teachers and students participating), identify the concrete step of the learning cycle in which the information is obtained, the place of observation, etc. 26 1.Integrate STEM content (and learning outcomes/goals) 1.Integrate STEM content (and learning outcomes/goals) 1.1.- Addressing key STEM learning outcomes/objectives Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Are the learning objectives presented, explained and discussed with the students? NE-Mentor Teacher Student Product: explanation/presentation digital format explanation/oral presentation Discussion of objectives Other_____________ Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment b. Do teachers and students share objectives and criteria for the achievement of competencies? NE-Mentor Teacher Student Product: Discussion of objectives Agreement on achievement criteria Register of agreements Other_____________ Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment c. The teacher asks students to clarify the nature or demands of the task, how to approach it and the product or outcome required NE-Mentor Teacher Student Open questions Closed questions Progress report on the task Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment d. When working, do students take into account the activity if their performance is goal-oriented? NE-Mentor Teacher Student Objectives are taken up again when the task is set Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Consider the objectives at some point during the task Other_____________ Virtual enviroment Students Mentors Artifact analysis sheet 1.2.- Interdisciplinarity of STEM content a. Do the teachers involved work as a team throughout the whole activity? NE-Mentor Teacher Student They plan together They hold meetings for follow-up Several teachers are in the classroom together Other:_____________ Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment b. Is the theoretical and practical knowledge of different subjects necessary in the activity? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 28 c. Are the relationships between the disciplines explained in order to carry out the activity? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: d. Do the students use the knowledge of the different disciplines when carrying out the activity? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: e. Do students discuss and reflect on how all this interdisciplinary knowledge helps them in solving the tasks? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: f. Are assessment criteria developed among the STEM teachers? NE-Mentor Teacher Student Yes Only in some activities/tasks No Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 29 g. Is the process co-evaluated by the Team of STEM teachers? NE-Mentor Teacher Student Yes Only in some activities/tasks No Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 30 2. Reflect STEM learning design principles (and social constructivist views of learning) . Reflect STEM learning design principles (and social constructivist views of learning) 2.1.- Working in the classroom with problems and challenges Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Are the problems raised connected to the real world? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: b. Is research linked to the real world encouraged? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME (explanation) NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: c. Is technological or engineering design encouraged? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 31 31 d. Are the problems or challenges presented (by teacher/students) regarding the environment or with previous ideas and knowledge of the students? NE-Mentor Teacher Student Previous ideas are explored A case is made for discussion Other____________ Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet e. Is input from all team members on the problem encouraged? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: f. Are they encouraged to build the solution using new knowledge? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: g. Are they encouraged to bring their vision of how to bring this problem to real life? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 32 32 2.2.- Reflecting social constructivist views on learning a. Is cooperative learning promoted? (search for definition) NE-Mentor Teacher Student They plan together They hold meetings for follow-up They discuss and make decisions Other:_____________ Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet b. Does the teacher act as a facilitator of the joint educational process among the students? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: c. Does the teacher encourage the active commitment of each and every member of the classroom? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: d. What tools do you use to foster cooperation? NE-Mentor Teacher Student Communication tools Information sharing tools Tools for building together Example of each tool used Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 33 e. Does the team have autonomy in decisions about its own learning process? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: f. Is team assessment encouraged as a pillar of the constructivist learning process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: g. Are channels encouraged for different work teams to share the results of their activities or work? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 2.3- Stimulating creativity to offer more than one solution to problems. a. Is the classroom a safe environment in which students can express themselves without fear of making mistakes? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: e. Does the team have autonomy in decisions about its own learning process? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: f. Is team assessment encouraged as a pillar of the constructivist learning process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: g. 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME Are channels encouraged for different work teams to share the results of their activities or work? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 2.3- Stimulating creativity to offer more than one solution to problems. a. Is the classroom a safe environment in which students can express themselves without fear of making mistakes? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 34 34 b. Are failure and error conceptualised as an indispensable part of the learning process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: c. Are there problems, questions or activities that are open enough for students to have room to explore their own insights? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: d. Are channels open for all students to present and exchange their ideas and to stimulate creativity? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: e. Is the classroom a space for experimentation in which students are encouraged to adapt to different situations, allowing them to seek their own answers? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: f. Does the teacher encourage critical thinking by questioning the origin of his or her thoughts or ideas? NE-Mentor Teacher Student Always Most of the time Sometimes Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers 35 Hardly ever Never Explanation of any activity which demonstrate it: Students Mentors Artifact analysis sheet g. Do the activities motivate students to explore new knowledge for themselves? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: h. Are channels encouraged for different work teams to share the results of their activities or work? NE-Mentor Teacher Student Yes Only in some activities/tasks No Justification Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of any activity which demonstrate it: 36 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3.1. Feedback about a process Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Who gives the students feedback on the different tasks (individual teacher/Team of STEM teachers)? NE-Mentor Teacher Student Individual teacher Team of teachers Other_____________ Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment b. Is there feedback on learning achievements between teacher and students throughout the process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment c. Does the feedback provided give students information on their progress towards the objectives set? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Classroom Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Virtual enviroment d. Does the teacher give students opportunities to plan, stop and think, and review the tasks and NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 37 the process followed to solve them? Students Mentors Artifact analysis sheet e. Does the feedback recommend contents and strategies to improve the work or the evidence presented? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet f. 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME Does the teacher provide task- centerd feedback both in a clear and reasoned manner and in good time so that it can be used to improve successive tasks or learning cycles? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet g. Is formative feedback used which involves the inclusion of assessments with cognitive and metacognitive intent? (add examples on virtual environment observation sheet)? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 3.2 Feedback related to self-regulation a. Does the teacher provide his or her students with the correction criteria for the assessment of their own work? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students 38 Mentors Artifact analysis sheet b. Does the teacher convey the assessments to the students so that they can be used to improve learning autonomy? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet c. Does the feedback propose strategies to improve learning? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet d. Does the teacher encourage contexts of assessment (time of achievement) that allow students to generalise the knowledge acquired? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 39 e. Does the feedback provided suggest in-depth information so that the student can continue to progress autonomously in the future? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 40 4.- Facilitate peer-assessment (improve learning by activating students as instructional resources for one another Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Is there peer collaboration for the "training" process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet b. Is there peer collaboration for the assessment process? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet c. Does the teacher enable students to use the correction criteria to evaluate the work of their peers? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 4.- Facilitate peer-assessment (improve learning by activating students as instructional resources for one another 41 41 d. Are rubrics provided to students from the start to facilitate the assessment of their peers' work? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet e. Do students provide feedback on the learning process to their peers? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet f. Do students provide feedback on learning achievements to their peers? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet g. Does the teacher stimulate group discussion and reflection? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet h. 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME Does a group assignment correction and discussion take place? NE-Mentor Teacher Student Yes Only in some activities/tasks Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews 42 No Teachers Students Mentors Artifact analysis sheet i. In the proposed assessment: are there group grades? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 43 43 5.- Facilitate self-assessment (improve learning by activating students as owners of their own learning Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Do teaching staff encourage students to be actively involved in the assessment according to its characteristics and purposes? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet b. Is the student aware of the meaning of assessment in his/her learning process? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet c. Does the teacher encourage students to be aware of the learning results as well as help them decide to introduce changes and improvements in their learning? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 44 d. Does the teacher encourage the autonomy of the students to analyse and discuss new forms of assessment? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet e. Are assessment criteria and/or rubrics provided to students from the beginning to facilitate the assessment of their own work? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Attach a sample rubric Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet f. Are students competent to make valuable judgements about their own learning? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Muy competente … Explanation of the teacher's opinion: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet Explanation of the student's opinion: g. Do students identify what they have learned? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet h. Does the student identify what learning he or she should have accomplished? NE-Mentor Teacher Student Always Most of the time Sometimes Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews 45 Hardly ever Never Teachers Students Mentors Artifact analysis sheet i. Does the student attribute the learning results to his/her own causes such as effort, responsibility, and recognise that he/she can modify them in order to learn or improve his/her learning? NE-Mentor Teacher Student Attributes results to own causes Attributes results to external causes as a way of correcting the teacher, involvement of other colleagues... Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 46 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success Indicator: whom do we ask?- Informant What do we collect? Examples NOTES/ Collect through instrument a. Is student participation encouraged so as to contribute ideas in discussions and decision making? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet b. Is the activity aimed at making the result of everyone's work necessary for its achievement? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet c. Are students involved in all moments of the session (planning, explaining, executing, building, analysing, improving, etc.)? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 6. 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success 47 d. Do students support each other? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet e. Does everyone keep up by helping each other? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet f. Do they explain to each other how to do the task? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet g. Does everyone take responsibility for their work being integrated into the task? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet h. Are task sequences coordinated and shared during the work? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers 48 Never Students Mentors Artifact analysis sheet i. When there are coordination difficulties or disagreements in the way the task is solved, how is the conflict settled? (interview and specify for instruments) NE-Mentor Teacher Student Teacher mediation Collective resolution as a group (Resolución colectiva como grupo) Other_____________ Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet j. Did students analyse and reflect on whether they had achieved the goals? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet k. Do students reflect on the acquisition and advancement of their STEM skills? 6. DATA COLLECTION INSTRUMENTS AND RECOMMENDATIONS FOR THEIR IMPLEME NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet l. Do the STEM teaching staff jointly assess their students' acquisition of STEM skills? NE-Mentor Teacher Student Yes Only in some activities/tasks No Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 49 m. Are learning outcomes consistent with intentions? NE-Mentor Teacher Student Always Most of the time Sometimes Hardly ever Never Explanation of any activity which demonstrate it: Classroom observation record sheet Digital assessment tools Record Sheets Interviews Teachers Students Mentors Artifact analysis sheet 50 2.6.1. SELF-ADMINISTERED CORE ATS-STEM LEARNING COMPETENCES ONLINE QUESTIONNAIRE The questionnaire is an information collection technique that allows for obtaining information from a large number of participants in the field of education. This instrument can be defined as "... a structured technique that allows for the quick and abundant collection of information through a series of oral or written questions that an interviewee must answer with regard to one or more variables to be measured" (Albert, 2007, p.115). An electronic questionnaire was designed for the assessment of core ATS-STEM learning competences. This questionnaire is presented in the form of a 5-point lickert scale from very poor (1) to excellent (5) competence for pilot school students from all European countries participating in the pilot to cover before the start of the ATS-STEM project and again after the end 52 53 54 55 SEND 2.6.2. CLASSROOM OBSERVATION PROTOCOL Five classroom observations will be made in each of the learning cycles of the ATS-STEM projects, one in each of the steps of the cycle (defining a real world problem, finding solution(s), trialing solution(s), assessing solution(s) and discussing solution(s)). In the case of the classroom observation template, data will be collected both for the identification of the school (educational level, number of teachers and students participating...) and for the specific observation (step in which the observation is made, duration of the observation, place in which the observation is made... To support the observation, this sheet has been prepared based on the categories and subcategories of analysis explained in section 2.3. Information will be collected for each of the indicators (first column) indicating what is collected for each of the elements and providing specific examples that will later help you to relate this information to the collection through any of the other instruments. To facilitate data collection, a colour code has been used in which the indicator of each sub-dimension to be observed is related to the collection of specific information for it. In addition, a code is added to clarify and complete various issues observed that may help in the national report. To facilitate data collection, a colour code has been used in which the indicator of each sub-dimension to be observed is related to the collection of specific information for it. In addition, a code is added to clarify and complete various issues observed that may help in the national report. SEND SEND 56 56 Classroom observation sheet This instrument has been designed to document everything that happened in the classroom sessions developed during the ATS-STEM pilot. In this way, we will obtain evidence on the dynamics of interaction in the process of knowledge creation, the roles adopted by teachers and students and the difficulties of applying the programme with special attention to the use of digital assessment through a strategy of formative assessment. (i) General contextualization Date dd/mm/yy Country Name of the school Group /Level class This instrument has been designed to document everything that happened in the classroom sessions developed during the ATS-STEM pilot. In this way, we will obtain evidence on the dynamics of interaction in the process of knowledge creation, the roles adopted by teachers and students and the difficulties of applying the programme with special attention to the use of digital assessment through a strategy of formative assessment. (i) General contextualization Date dd/mm/yy Country Name of the school Group /Level class 57 Number of teachers Number of students (ii) Metadata Learning cycle title: dd/mm/yy Step: Orientation; Conceptualization; Research; Conclusion; Discussion Lesson Activity Lesson number Reference to the learning cycle (Add code) Linked to the observation in the classroom No: Number of teachers Number of students Duration of observation Place of observation 58 1.Integrate ATS-STEM content (and learning outcomes/goals) 1.1.- Addressing core ATS-STEM learning competences(learning outcomes/objectives a) Are the learning objectives presented, explained and discussed with the students? b) Do teachers and students share objectives and criteria for the achievement of competencies? c) The teacher asks students to clarify the nature or demands of the task, how to approach it and the product or outcome required d) When working, do students take into account the activity if their performance is goal-oriented? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. 1 Explanation/presentation digital format 2 Explanation/oral presentation 3 Discussion of objectives 4 Other: _______________ 5 Discussion of objectives 6 Agreement on achievement criteria 7 Register of agreements 8 Other: _______________ 9 Open questions 10 Closed questions 11 Progress report on the task 12 Objectives are taken up again when the task is set 13 Consider the objectives at some point during the task 14 Other_____________ a.1. c.8 d.12 . . . Note of a.1. Classroom observation sheet Note of d.12. 1.Integrate ATS-STEM content (and learning outcomes/goals) 59 1.2.- Interdisciplinarity of ATS-STEM content a) Do the teachers involved work as a team throughout the whole activity? b) Is the theoretical and practical knowledge of different subjects necessary in the activity? c) Are the relationships between the disciplines explained in order to carry out the activity? d) Do the students use the knowledge of the different disciplines when carrying out the activity? e) Do students discuss and reflect on how all this interdisciplinary knowledge helps them in solving the tasks? f) Are assessment criteria developed among the ATS-STEM teachers? g) Is the process co-evaluated by the Team of ATS-STEM teachers? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. 1 They plan together 2 They hold meetings for follow-up 3 Several teachers are in the classroom together 4 Other: _______________ 5 Always 6 Most of the time 7 Sometimes 8 Hardly ever 9 Never 10 Yes 11 Only in some activities/tasks 12 No a.1. e.8 g.12 . . . 1.2.- Interdisciplinarity of ATS-STEM content Do the teachers involved work as a team throughout the whole activity? 60 2. Reflect ATS-STEM learning design principles (and social constructivist views of learning) 2.1.- Working in the classroom with problems and challenges a) Are the problems raised connected to the real world? b) Is research linked to the real world encouraged? (explanation) c) Is technological or engineering design encouraged? d) Are the problems or challenges presented (by teacher/students) regarding the environment or with previous ideas and knowledge of the students? e) Is input from all team members on the problem encouraged? f) Are they encouraged to build the solution using new knowledge? Are they encouraged to bring their vision of how to bring this problem to real life? g) Are they encouraged to bring their vision of how to bring this problem to real life? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. Classroom observation sheet 1 Yes 2 Only in some activities/tasks 3 No 4 Previous ideas are explored 5 A case is made for discussion 6 Other____________ 7 always 8 most of the time 9 Sometimes 10 Hardly ever 11 Never 12 Yes 13 Only in some activities/tasks 14 No a.1. e.8 g.12 . . . M learning design principles (and social constructivist views of learning) 2. Reflect ATS-STEM learning design principles (and social constructivist views of learning) . Reflect ATS-STEM learning design principles (and social constructivist vi 61 2.2.- Reflecting social constructivist views on learning a) Is cooperative learning promoted? (search for definition) b) Does the teacher act as a facilitator of the joint educational process among the students? c) Does the teacher encourage the active commitment of each and every member of the classroom? d) What tools do you use to foster cooperation? e) Does the team have autonomy in decisions about its own learning process? f) Is team assessment encouraged as a pillar of the constructivist learning process? g) Are channels encouraged for different work teams to share the results of their activities or work? Indicator What do we collect? Examples Notes a. b. c. d. e. f. g. 1 They plan together 2 They hold meetings for follow-up 3 They discuss and make decisions 4 Other: ___________ 5 Always 6 Most of the time 7 Sometimes 8 Hardly ever 9 Never 10 Communication tools 11 Information sharing tools 12 Tools for building together 13 Yes 14 Only in some activities/tasks 15 No a.1. e.8 g.12 . . . 62 2.3- Stimulating creativity to offer more than one solution to problems. a) Is the classroom a safe environment in which students can express themselves without fear of making mistakes? b) Are failure and error conceptualised as an indispensable part of the learning process? c) Are there problems, questions or activities that are open enough for students to have room to explore their own insights? d) Are channels open for all students to present and exchange their ideas and to stimulate creativity? e) Is the classroom a space for experimentation in which students are encouraged to adapt to different situations, allowing them to seek their own answers? f) Does the teacher encourage critical thinking by questioning the origin of his or her thoughts or ideas? g) Do the activities motivate students to explore new knowledge for themselves? Classroom observation sheet h) Are channels encouraged for different work teams to share the results of their activities or work? Indicator ¿Qué recogemos?* Examples Notes a. b. c. d. e. f. g. h. 1 Always 2 Most of the time 3 Sometimes 4 Hardly ever 5 Yes 6 Only in some activities/tasks 7 No a.1. e.8 g.12 . . . 2.3- Stimulating creativity to offer more than one solution to problems. 63 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3.1. Feedback about a process 3.1. Feedback about a process a) Who gives the students feedback on the different tasks (individual teacher/Team of ATS-STEM teachers)? b) Is there feedback on learning achievements between teacher and students throughout the process? c) Does the feedback provided give students information on their progress towards the objectives set? d) Does the teacher give students opportunities to plan, stop and think, and review the tasks and the process followed to solve them? e) Does the feedback recommend contents and strategies to improve the work or the evidence presented? f) Does the teacher provide task-centerd feedback both in a clear and reasoned manner and in good time so that it can be used to improve successive tasks or learning cycles? g) Is formative feedback used which involves the inclusion of assessments with cognitive and metacognitive intent? (add examples on virtual environment observation sheet)? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. 1 Individual teacher 2 Team of teachers 3 Other: ___________ 4 Yes 5 Only in some activities/tasks 6 No 7 always 8 most of the time 9 Sometimes 10 Hardly ever 11 Never a.1. e.8 g.12 . . . 64 3.2. Feedback related to self-regulation 3.2. Feedback related to self-regulation a) Does the teacher provide his or her students with the correction criteria for the assessment of their own work? b) Does the teacher convey the assessments to the students so that they can be used to improve learning autonomy? c) Does the feedback propose strategies to improve learning? d) Does the teacher encourage contexts of assessment (time of achievement) that allow students to generalise the knowledge acquired? e) Does the feedback provided suggest in-depth information so that the student can continue to progress autonomously in the future? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. 1 Yes 5 Only in some activities/tasks 6 No 4 always 5 Most of the time 6 Sometimes 7 Hardly ever 8 Never a.1. e.8 g.12 . . . 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner Notes (personal input to help clarify what is happening in the observation). 65 4.- Facilitate peer-assessment (improve learning by activating students as instructional resources for one another a) Is there peer collaboration for the "training" process? b) Is there peer collaboration for the assessment process? c) Does the teacher enable students to use the correction criteria to evaluate the work of their peers? d) Are rubrics provided to students from the start to facilitate the assessment of their peers' work? e) Do students provide feedback on the learning process to their peers? f) Do students provide feedback on learning achievements to their peers? g) Does the teacher stimulate group discussion and reflection? h) Does a group assignment correction and discussion take place? i) In the proposed assessment: are there group grades? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. i. 1 Yes 2 Only in some activities/tasks 3 No 4 always 5 Most of the time 6 Sometimes 7 Hardly ever 8 Never a.1. e.8 g.12 . . . 4.- Facilitate peer-assessment (improve learning by activating students 66 5.- Facilitate self-assessment (improve learning by activating students as owners of their own learning a) Do teaching staff encourage students to be actively involved in the assessment according to its characteristics and purposes? b) Is the student aware of the meaning of assessment in his/her learning process? c) Does the teacher encourage students to be aware of the learning results as well as help them decide to introduce changes and improvements in their learning?? d) Does the teacher encourage the autonomy of the students to analyse and discuss new forms of assessment? e) Are assessment criteria and/or rubrics provided to students from the beginning to facilitate the assessment of their own work? f) Are students competent to make valuable judgements about their own learning? g) Do students identify what they have learned? h) Does the student identify what learning he or she should have accomplished? i) Does the student attribute the learning results to his/her own causes such as effort, responsibility, and recognise that he/she can modify them in order to learn or improve his/her learning? Indicator What do we collect? 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. i. 1 always 2 Most of the time 3 Sometimes 4 Hardly ever 5 Never 1 Yes 2 Only in some activities/tasks 3 No 9 Very poor competence 10 Not good competence 11 All right 10 Good competence 11 Excellent competence 12 Attributes results to own causes 13 Attributes results to external causes as a way of correcting the teacher, involvement of other colleagues... a.1. e.8 g.12 . . . 5.- Facilitate self-assessment (improve learning by activating students as owners of their own learning improve learning by activating students as owners of their own learning 67 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting clarify the meaning of success a) Is student participation encouraged so as to contribute ideas in discussions and decision making? b) Is the activity aimed at making the result of everyone's work necessary for its achievement? c) Are students involved in all moments of the session (planning, explaining, executing, building, analysing, improving, etc.)? d) Do students support each other? e) Does everyone keep up by helping each other? f) Do they explain to each other how to do the task? g) Does everyone take responsibility for their work being integrated into the task? h) Are task sequences coordinated and shared during the work? i) When there are coordination difficulties or disagreements in the way the task is solved, how is the conflict settled? (interview and specify for instruments) j) Did students analyse and reflect on whether they had achieved the goals? k) Do students reflect on the acquisition and advancement of their core ATS-STEM learning competences? l) Do the STEM teaching staff jointly assess their students' acquisition of ATS-STEM learning competences? m) Are learning outcomes consistent with intentions? Indicator What do we collect? Examples (to be included in those indicators where examples from that observation session can be reflected) Notes (personal input to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. i. j. k. l. 1 Yes 2 Only in some activities/tasks 3 No 4 always 5 Most of the time 6 Sometimes 7 Hardly ever 8 Never 9 teacher mediation 10 Collective resolution as a group 11 Other_________________ a.1. e.8 g.12 . . . 68 68 m. 69 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL The record sheets are designed to collect information on the interaction, communication, collaboration and reflection dynamics generated through the digital assessment tools implemented. As with the classroom observation sheets, contextualisation data and data from the specific learning cycle in which the observation of the virtual environment is carried out are collected. In this case, the same dimensions and indicators have been taken but we will focus the data collection on the type of tools used for each of the functions and on how the implementation and use of the digital assessment tools support the processes. For the analysis of interaction, communication, collaboration, and reflection dynamics generated through the digital assessment tools implemented (such as digital portfolios). These data will allow us to understand the role and possibilities of digital assessment in the implementation of the ATS-STEM program. (I) General contextualization Date dd/mm/yy Country Name of the school Group /Level class Number of teachers Number of students (II) Metadata Learning cycle title: dd/mm/yy Step: Orientation; Conceptualization; Research; Conclusion; Discussion Lesson Activity Lesson number Reference to the learning cycle (Add code) Linked to the observation in the classroom No: Number of teachers Number of students communication, collaboration, and reflection dynamics generated through the digital assessment tools implemented (such as digital portfolios). These data role and possibilities of digital assessment in the implementation of the ATS-STEM program. 72 1. Integrate STEM content (and learning outcomes/goals) 1.1. Addressing core ATS-STEM learning competences/ learning outcomes/objectives Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 1. 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL Integrate STEM content (and learning outcomes/goals) 73 1.2.- Interdisciplinarity of ATS-STEM content Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 74 Reflect ATS-STEM learning design principles (and social constructivist views of learning) .- Working in the classroom with problems and challenges cator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 2. Reflect ATS-STEM learning design principles (and social constructivist views of learning) 2.1.- Working in the classroom with problems and challenges 75 F4.2. Co-assessment F4.3. Self-assessment 76 2.2.- Reflecting social constructivist views on learning Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL b. c. d. e. f. g. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 77 2.3- Stimulating creativity to offer more than one solution to problems. Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 2.3- Stimulating creativity to offer more than one solution to problems. 78 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3.1. Feedback about a process Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow- up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 79 F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment 80 3.2 Feedback related to self-regulation Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 81 Facilitate peer-assessment (improve learning by activating students as instructional resources for one another cator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). . . . . . . . T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . 2.6.3. VIRTUAL ENVIRONMENT OBSERVATION PROTOCOL Note of a.T1.a.F1.1 Note of 82 5.- Facilitate self-assessment (improve learning by activating students as owners of their own learning) Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. i. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of learning by activating students as owners of their own learning) 83 6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success) Indicator (I) Tool (T) Function (F) Examples (They will be collected in those indicators in which examples of that observation session can be reflected) Notes (personal contributions to help clarify what is happening in the observation). a. b. c. d. e. f. g. h. i. T1 Synchronous communication tools: a)________________________________ T2 Asynchronous communication tools: _________________________________ T3 Tools for collaboration: _________________________________ T4 Tools to create T5 Tools for viewing and sharing products T6 Tools to evaluate F1 Communication 1.1. Between the whole group-po 1.2. Between work teams 1.3. Individual (between pairs) F2 Collaboration F2.1. Planning together F2.2. Holding meetings for follow-up F2.3. Discussing and making decisions F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other: _______________ F2.4. Other F3 Feedback F3.1. Continuous F3.2. Self-regulation F4 Assessment F4.1. Peer review F4.2. Co-assessment F4.3. Self-assessment Ia.T1.a.F1.1 . . . Note of a.T1.a.F1.1 Note of 84 2.6.4. ATS-STEM LEARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) 2.6.4. ATS-STEM LEARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) ARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) ARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) 2.6.4. ATS-STEM LEARNING CYCLES DESIGN (for the planning of data collection for each ATS-STEM Project) 85 . 85 2.6.5. ARTIFACT ANALYSIS PROTOCOL The next instrument designed is the artifact analysis sheet. By "artifact" we mean any elaboration or product created by the student in each of the ATS-STEM projects. This includes reflections or comments on the e-portfolio, models, multimedia or hypermedia creations, podcasts, videos etc. This instrument will be used to evaluate three different artefacts per ATS-STEM project, in which variety in the levels of academic performance (low, medium and high) should be appreciated. Through this production, the achievement of the eight core ATS-STEM learning competences will be analysed as well as the formative digital assessment strategies carried out during the product creation period. It is therefore important to bear in mind that the aim of this instrument is not to judge the productions as a final product, but rather the whole process that has led to their creation. Artifact analysis sheet For the analysis of each artefact produced by the students, this analysis sheet should be used. It includes (I) information on the contextualization of the task, (II) metadata on the artefact to interpret it in relation to the design of the ATS-STEM project, (III) observable elements linked to the formative assessment tasks and the core STEM Competencies. (I) General contextualization Date dd/mm/yy Country Name of the school Group /Level class Number of teachers Number of students (II) Metadata Learning cycle title/ATS-STEM project name: Step: Orientation; Conceptualization; Research; Conclusion; Discussion Goal of the Lesson Description of the activity Reference to the learning cycle (Add code) Linked to the observation in the classroom No: Number of teachers Number of students Artifact type e-portfolio; Reflection; discussion forum; digital presentation; video interactive video; podcast; other ____________________ 88 Core ATS-STEM learning competencies involved In this table you must indicate which of the competences included in the ATS-STEM teaching methodology are involved in the development of this product, evaluating their level of competence. Not all of them must be involved, so select those whose achievement is a priority objective for this product. Also, remember that the most important thing is that you provide us with a detailed justification and description. Competence involved if applicable Level of achievement Justification and description of the level of achievement Problem Solving excellent Medium low Innovation and creativity excellent Medium low Communication excellent Medium low Critical Thinking excellent Medium low Meta – cognitive skills excellent Medium low Collaboration excellent Medium low 89 Strategies of formative assessment Strategies of formative (digital) assessment carried out during the process of elaboration of this product. Self – regulation excellent Medium low Disciplinary competences excellent Medium low Self – regulation excellent Medium low Disciplinary competences excellent Medium low Strategies of formative assessment Strategies of formative assessment 90 Formative assessment tasks For the analysis of the artifact, we must situate ourselves in the concrete lesson of the cycle and the corresponding step and for this analysis a registration table was elaborated that follows the sequence of the corresponding dimensions and indicators. For the analysis of the artifact, we must situate ourselves in the concrete lesson of the cycle and the corresponding step and for this analysis a registration table was elaborated that follows the sequence of the corresponding dimensions and indicators. 1. Integrate ATS.STEM content (and learning outcomes/goals) 1.1 Addressing core ATS-STEM learning competences/ learning outcomes/objectives Are the objectives of this device guided by the learning objectives? 1.2 Interdisciplinarity of ATS-STEM content The production of this device requires theoretical and practical knowledge of different areas of knowledge? 2. Reflect ATS-STEM learning design principles (and social constructivist views of learning) 2.1. Working in the classroom with the real-world problems and challenges 91 The artifact is linked to the real world: works with real world data and responds to real problems 2.2.- Reflecting social constructivist views on learning In the production of this artifact, can the vision of learning as a process of social construction be appreciated? 2.3 - Stimulating creativity to offer more than one solution to problems The approach to making the artefact allows for the creativity of the students 3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner 3.1 Feedback about a process ing the learner to use effective feedback focused on the learning outcome/goal in a timely manner 92 In case it is possible to identify it: how was the feedback on the process of elaboration of the artefact 3.2 Feedback related to self-regulation In case it is possible to identify it: how was the feedback for the self-regulation of the student in the process of elaboration of the artefact 4. Facilitate peer-assessment: Improve learning by activating students as instructional resources for one another Contributions/comments have been made by colleagues/members of other groups during the development of the device and in the final product 5. Facilitate self-assessment: improve learning by activating students as owners of their own learning The student/group has carried out some process of self-assessment that has resulted in the improvement of the device 93 93 6. 2.6.6. INTERVIEW PROTOCOL Finally, the last of our instruments are the interviews that will be conducted with teachers, students and mentors of each ATS-STEM project. Finally, the last of our instruments are the interviews that will be conducted with teachers, students and mentors of each ATS-STEM project. The The interview is the most widely used social research technique and it allows us to collect subjective information about events, beliefs, attitudes, values etc (Aguiar & Barroso, 2015, p. 79). In this sense, our instrument is designed to collect information about how the process of implementation of the ATS-STEM projects went, what difficulties arose in their implementation and how they were solved. Furthermore, by conducting interviews we will complement the information obtained both through the questionnaires and through observation in the classroom and virtual environment. This will be performed in a way that will allow us to obtain more information, give confidence to the data obtained and deepen in a relaxed context how the participants of the ATS-STEM projects perceive the benefits in the use of digital assessment for the integration of ATS-STEM projects. Regarding the use of this instrument, it is important that in its application the interviews with teachers and students are carried out, each of them, in a group way, selecting a group of between three and six participants of the same ATS-STEM project in the case of the groups of teachers and three students of different academic performance level and gender. As for the interview with mentors, these can be carried out individually. The first one - interviews with teachers - attempts to identify the global perception of teachers/students and mentors about the experience of the ATS2020 model. The second of these - interviews with students - aims to find out the perception of each agent profile in relation to the indicators, sub-dimensions and dimensions of analysis. The third, -interview with mentors-, focuses on the perceptions of the main difficulties experienced by the teaching staff and the information on how far the digital assessment training strategies help to develop the eight core ATS-STEM learning competences. Formative assessment tasks Help to Elicit evidence of learning This artifact allows for the identification of whether the learning outcomes are consistent with the stated intentions. 94 2.6.6.1. TEACHING TEAMS [2.3 (f)] As a teacher, do you facilitate students' critical questioning of the origin of their ideas and perceptions about the issues worked on in the classroom in ATS-STEM projects? [2.3 (f)] As a teacher, do you facilitate students' critical questioning of the origin of their ideas and perceptions about the issues worked on in the classroom in ATS-STEM projects? [3.1 (b) (e) (f)] As a teacher, do you give feedback to students throughout the process? Does the feedback contain information that allows students to improve both the task they are doing and their future learning processes? [3.1 (b) (e) (f)] As a teacher, do you give feedback to students throughout the process? Does the feedback contain information that allows students to improve both the task they are doing and their future learning processes? [3.1 (g)] As a teacher, do you give formative feedback that aims to make students aware of how they are doing their learning and to improve future processes? [3.2 (c) (d) (e)] Did the feedback given during the projects provide sufficient information to the students to improve their strategies and autonomy in learning, as well as to generalise the knowledge acquired? [4 (c) (d) (h) (i)] Is peer assessment promoted by providing assessment criteria and rubrics for group correction of tasks? Are group ratings provided in addition to individual ratings? [4 (g)] In your activity as a teacher, do you try to stimulate debate among your students through group reflection? [5(b) (c)] In your educational proposal, have you made sure that students are aware of what they are expected to achieve with their learning, so that they have enough information to introduce changes in their own learning process? In relation to assessment, have you taken into account the students' role in the whole learning process? In your training proposal, do you consider that you have encouraged students to take into account both the learning results and the meaning of the assessment so that they can introduce changes that will improve their learning? [5(e)] Have you provided criteria or headings for students to self-assess their learning? (f) (i)] Have students been responsible for their learning and have they been able to assess their learning [6 (c) (e)] During the implementation of ATS-STEM projects, students actively participate in all phases of the session? And, do they support each other in achieving the learning objectives? 2.6.6.1. TEACHING TEAMS [1.1 (a) and (b)] Prior to and during the different phases of the ATS-STEM project, are the learning objectives and the achievement of the core ATS-STEM learning competences shared and discussed with the students? [1.1 (d) (c)] During the implementation of the proposed activities, do the students take into account the proposed objectives and, as teachers, do you ask for evidence of it (clarification on the demands of the task, how to approach them and/or required outcome)? [1.2 (a) (f) (g)] In relation to your organisation, have you worked as a team throughout the project including assessment (Elaboration of criteria and co-assessment among the team of teachers? [1.2 (b) (c) (d)] During the implementation of the ATS-STEM projects, are the relationships between the different subjects explained and are students encouraged to link their theoretical and practical knowledge to carry out the different activities proposed? [2.1 (a) (d) (f) (g)] Are the problems encountered related to the real world and do the students' previous knowledge lead to the construction of new knowledge applicable to the real world? [2.1 (c)] In the proposal for ATS-STEM projects, do you encourage technological or engineering design so that students review the proposed solutions through a cyclical process of improvement? [2.2 (a) and (d)] Is cooperative learning promoted? What tools are used to promote such learning? [2.1 (e) and 2.2 (c)(e)(g) and 2.3 (h)] Is team learning promoted by providing tools that facilitate communication among students and empower teams to make decisions? [2.1 (e) and 2.2 (c)(e)(g) and 2.3 (h)] Is team learning promoted by providing tools that facilitate communication among students and empower teams to make decisions? [2.3 (a) (b)] Is the classroom presented as an environment in which to express oneself freely and is error accepted as an important part of learning? [2.3 (a) (b)] Is the classroom presented as an environment in which to express oneself freely and is error accepted as an important part of learning? [2.3 (c) (d) (e) (g)] Are the activities proposed broad enough for all students to contribute their insights? Do these activities empower students to find solutions and exchange ideas with their peers? [2.3 (c) (d) (e) (g)] Are the activities proposed broad enough for all students to contribute their insights? Do these activities empower students to find solutions and exchange ideas with their peers? 2.6.6.2. STUDENT GROUPS [1.1 (a) and (b)] Prior to and during the various phases of the ATS-STEM project, have your teachers explained to you and discussed what you are expected to learn and what skills you should acquire through your participation in the project? [1.1 (d) (c)] While you are carrying out the activities proposed by your teachers, do you think again that you are expected to learn from this activity? [1.2 (b) (c) (d) (e)] During your participation in the ATS-STEM projects, have you been shown the relationships and the need to use the theoretical and practical knowledge of the different subjects in order to carry out the proposed activities? Have you been aware of how this knowledge of the different subjects has helped you to solve the activities? [2.1 (a) (b) (d) (f) (g)] Are the activities proposed to you linked to the real world and to your previous knowledge? Do you think that you obtained new knowledge allowing you to improve a real situation? [2.2 (a) and (d)] Is cooperative learning promoted? What tools are used to promote such learning? [2.1 (e) and 2.2 (b) (c)(e) (g) and 2.3 (h)] Is team learning promoted by providing tools that facilitate communication between students and empowering teams to make decisions? [2.1 (e) and 2.2 (b) (c)(e) (g) and 2.3 (h)] Is team learning promoted by providing tools that facilitate communication between students and empowering teams to make decisions? [2.2 (a) (b) (d) (e)] Do teachers try to get you to work as a team, give you autonomy to do so and provide you with tools to do so? [2.2 (a) (b) (d) (e)] Do teachers try to get you to work as a team, give you autonomy to do so and provide you with tools to do so? [2.2 (c) (f)] Does the teacher try to get all team members actively involved and, in addition to individual assessment, is there assessment of teamwork? hers encourage you to share the results of your activities with your colleagues? [2.2 (g)] Do teachers encourage you to share the results of your activities with your colleagues? [2.3 (a) (b)] Do you think the classroom is a safe environment where you can express yourself freely and where mistakes are accepted as an important part of learning? 2.6.6.1. TEACHING TEAMS [6 (c)] In the event of disagreements between students as a whole or with work teams, what strategies have you used and what measures have you implemented to help resolve the conflict? [6 (c)] In the event of disagreements between students as a whole or with work teams, what strategies have you used and what measures have you implemented to help resolve the conflict? [6. (l)] Has the team of ATS-STEM teachers jointly evaluated the acquisition of core ATS-STEM learning competences by your students? [6. (l)] Has the team of ATS-STEM teachers jointly evaluated the acquisition of core ATS-STEM learning competences by your students? 96 96 2.6.6.2. STUDENT GROUPS [4 (i)] Did your assessment include group grades in addition to individual grades? [4 (e) (f)], Did you have the opportunity to provide feedback to your peers on the assignment and learning outcomes? [5 (a) (b) (c) (d)] During the course of the ATS-STEM projects, have you felt involved and able to influence your assessment process? In addition, have you had the opportunity to propose new forms of assessment? [5 (e)] Has the process provided you with criteria or rubrics for self-assessment of your learning? [5 (g) (h) (f) (i) and 6. (j)] During your participation in the ATS-STEM projects and considering the objectives proposed, have you been able to identify which ones you have achieved and which ones you have not? Do you consider yourself responsible for your learning? Are you able to make value judgements about your learning? [6 (b) (d) (e) (f) (g)] During your participation in the ATS-STEM projects, do you consider that teamwork has been necessary (support, help, explanations, collective responsibility and individual contribution to teamwork etc.). [6 (a) (i)] Is everyone encouraged to participate by contributing their ideas? And, when there are disagreements, how do you resolve them? [6 (a) (i)] Is everyone encouraged to participate by contributing their ideas? And, when there are disagreements, how do you resolve them? [6 (k)] Have you been aware of what core ATS-STEM learning competences you have acquired in the course of your ATS- ATS-STEM projects? 2.6.6.2. STUDENT GROUPS [2.3(c)(d)(e)(g)] Do you think that the activities proposed by teachers are broad enough for everyone to contribute their insights and ideas, and do these activities promote the exchange of ideas with your peers and the search for new solutions? [2.3(f)] Do teachers help you to think deeply and critically about the origin of your ideas concerning the issues worked on in the classroom in the ATS-STEM projects? [3.1 (b) (e) (f)] Have you received feedback throughout the process or only at certain times in relation to specific tasks? This feedback contained information that allowed you to improve the activity you were doing. Will it be useful for future learning? [3.1 (b) (e) (f)] Have you received feedback throughout the process or only at certain times in relation to specific tasks? This feedback contained information that allowed you to improve the activity you were doing. Will it be useful for future learning? [3.1 (c) (d)] Did the feedback you received from the teachers provide you with information to check your progress and rethink your approach to the activity? [3.1 (g)] Do you think that the feedback you received from the teachers made you aware of the way you learn and you think this will be useful for future learning? [3.1 (g)] Do you think that the feedback you received from the teachers made you aware of the way you learn and you think this will be useful for future learning? 97 97 [3.2 (c) (d) (e)] Did the feedback you received from your teachers during your participation in the ATS-STEM projects enable you to improve your learning strategies and become more autonomous in your learning process? [3.2 (c) (d) (e)] Did the feedback you received from your teachers during your participation in the ATS-STEM projects enable you to improve your learning strategies and become more autonomous in your learning process? [3.2 (a) (b)] Did the teacher provide you with correction criteria to evaluate your own work? Did the assessments he/she made of your work allow you to be more autonomous in your learning? [4 (a) (b) (c) (d) (g) (h)] During the ATS-STEM projects you participated in, was there collaboration with your peers throughout the process, including assessment? Did the faculty provide you with criteria for evaluating your peers' work? [4 (i)] Did your assessment include group grades in addition to individual grades? 2.6.6.3. MENTORS [1.2 (a) (b) (f) (g)] In relation to the organization of the team of teachers who have participated in each ATS-STEM project, do you consider that they have worked as a team during all the phases of the project including the assessment (elaboration of criteria and co-assessment)? [3.1 (a)] Who gives feedback to the students on the tasks presented (Individual teacher/ATS-STEM teaching team)? [3.1] Did the teachers show any difficulties in the feedback processes? Which ones? [3.1 and 3.2] Did the teachers require any specific support regarding the use of digital tools to carry out the formative assessment? -What have been the most frequent inquiries from teachers regarding the design of ATS-STEM projects? -What have been the requests for support from the teaching staff related to the use of digital tools for the formative assessment of students in ATS-STEM projects? 98 98 99 3. DATA ANALYSIS: BASIC SUPPORT FOR THE ANALYSIS OF QUALITATIVE DATA. In qualitative research you work with a lot of information, and it is important to be rigorous in the handling of the data and the interpretation made by the researcher. This must always be based on the evidence collected in the study about reality and not let the researcher's personal vision and beliefs prevail. To this end, qualitative methodology is based on the design of instruments and the triangulation of data, sources, and methods. In this research, the process steps were clearly established to help the analysis process to be developed equally by all field researchers. The data collected in the instruments follow the indicator structure, which was already established in their design, to answer the research questions. In the analysis of the data, it is necessary to follow the indicators and use the data without losing the relationship of each data with the source and the processes experienced in the studied reality. To this end, the instrument was constructed with categories, subcategories, indicators, what is collected, and what information collection instrument/source is involved. With this instrument for analysis, the researcher must read their data, interpret, and explain the result in each indicator, also indicating at what point in the teaching/learning process or if it is an artefact/observation/interview and who is responsible for it, student/teacher, in order to be able to contrast with the data. Literal texts will also be collected in quotation marks as evidence of interpretation, for example a textual fragment of an interview or a classroom observation. The process of analysis is carried out from the evidence and in a process of reduction in which explanation and detail is given, guided by the created system of indicators, subcategories, and categories. Working first on each indicator independently, after this analysis all the indicators of the subcategory to which they belong are interpreted and finally the interpretation of each category is made with the analyses of the subcategories of each category. When the whole process of analysis is complete, it is possible to arrive at a total interpretation and explanation of the studied reality that allows us to answer the research questions based on the scientific evidence on reality. 1º Working with indicators: In the system of indicators created, each one of them has a guideline "what we collect" to have criteria that help to determine the evidence more precisely. 2.7. DATA TRIANGULATION (INCLUDING EXAMPLES) The use of data triangulation in educational research is extremely important as a guarantee of quality, rigor and validity in the research process (Aguilar & Barroso, 2015). Triangulation as an assessment strategy in education increases the reliability of results and minimizes bias (Betrián Villas, Galitó Gispert, García Merino, Jové Monclús, Macarulla Garcia, 2013). To evaluate how the use of digital assessment strategies impact on an improvement of the implementation of ATS-STEM projects we need to obtain information through the collection of different types of data (quantitative and qualitative), through different methods (observation, field notes and interviews) and through different agents/participants in the research (teachers, students and mentors). The combination of quantitative and qualitative data are methods that complement each other and allow the strengths and weaknesses of each one to be used, comparing the information obtained and making it possible to check whether the same results are obtained, i.e. whether they converge or whether, on the contrary, they diverge and are contradictory. In this way we collect data: • Quantitative: through the questionnaire of self-assessment of core ATS-STEM learning competences of the students before and after the participation in the ATS-STEM projects to check how the self-assessment of these competences varies. • Qualitative: collected through the sheets of different methods: observation both in the classroom and in the virtual environment used to support the learning cycles and through the interviews carried out with the 3 agents participating in the projects. The fact of obtaining data from different agents provides a variety of perspectives on the same data which eliminates the bias of obtaining information from a single researcher. 100 3. DATA ANALYSIS: BASIC SUPPORT FOR THE ANALYSIS OF QUALITATIVE DATA. But this guideline is not the same for all the indicators because of the nature of the data that is being collected with the different instruments. There are several guidelines: 1. In the case that what is collected has specific aspects of the activity such as Explanation/presentation digital format Open questions They plan together Individual teacher Explanation/oral presentation Closed questions They hold meetings for follow-up Teacher´s team Discussion of objectives Progress report on the task Several teachers are in the classroom together Other_____________ Other_____________ Other_____________ 1. In the case that what is collected has specific aspects of the activity such as 101 101 Each option needs to be explained and examples given to illustrate all the strategies referred to in the indicator. Also compare which option appears most in the evidence. And in the case of "Others" collect them and indicate if they are new or like the options proposed. Each option needs to be explained and examples given to illustrate all the strategies referred to in the indicator. Also compare which option appears most in the evidence. And in the case of "Others" collect them and indicate if they are new or like the options proposed. 2º In the case that what is collected is focused on the activity frequency such as Allways Yes Most of the time Only in some activities/tasks Sometimes No Hardly ever Never In these situations, all the options can be explained by identifying the number of times it appears according to the number of evidence. Of those that are more present, examples would be given to illustrate the indicator. And in the case of "Only some activities/tasks" indicate some of the most frequent ones. In these situations, all the options can be explained by identifying the number of times it appears according to the number of evidence. Of those that are more present, examples would be given to illustrate the indicator. And in the case of "Only some activities/tasks" indicate some of the most frequent ones. 2nd Work with the subcategories For the work with subcategories the researcher has to relate the interpretations he made of his indicators to respond to that subcategory. The objective of this analysis is to characterize what was done in the action when the pilot experiences were implemented. Everything that appears in the interpretation of the indicators will respond to whether and how those elements referred to by the indicator appeared in the reality studied. To do this, they will have to relate it to the planning of the pilot experiences to see the learning goals that were intended and explain how the indicator shows us what happened. That is to say, if in the design of an activity "debate" was not considered, it cannot be stated in the analysis that there is no participation of the students in debates. The researcher has to review all the plans that were designed, and, in the analysis, he has to explain all the elements that were collected about the subcategory that were planned (how they were). 3rd Work with the categories 3rd Work with the categories 3) how digital tools have been used. 3rd Work with the categories The categories have been designed according to the dimensions of the ATS-STEM teaching methodology and reflect both a ATS-STEM teaching methodology and the formative assessment approach that can support STEM education. Therefore, now at this point in the analysis we contrast whether the action, when implementing the pilots, has developed according to the features that define the ATS-STEM teaching methodology and that are specified in the six categories. Each category includes the analysis of its subcategories, and these explanations and interpretations are worked on from the evidence collected with the qualitative instruments and from the different sources and methods. In explaining the six categories, the following points will be followed: 1) how the principles of ATS-STEM teaching methodology are applied 1) how the principles of ATS-STEM teaching methodology are applied 2) what forms of formative assessment have been put in place 2) what forms of formative assessment have been put in place 102 3) how digital tools have been used. 4) and what aspects can be consolidated or if it is necessary to look for ways to improve them. 103 4. NATIONAL REPORT: STRUCTURE AND FORMAT. CHAPTER 1. INTRODUCTION This section should include the following information: This section should include the following information: 1.1.1 Description of the structure of the country's education system and levels of education by age 1.1.1 Description of the structure of the country's education system and levels of education by ag Graphic presentation of the different stages and cycles and types of teaching as they are passed from one to another Graphic presentation of the different stages and cycles and types of teaching as they are passed from one to another See as an example the computer graphics of the Spanish educational system provided below See as an example the computer graphics of the Spanish educational system provided below See as an example the computer graphics of the Spanish educational system provided below STEM Education in the national education system See as an example the computer graphics of the Spanish educational system provided below STEM Education in the national education system D t il d d i ti f h STEM d ti i t l t d d/ h d i th t ' d ti l t d STEM Education in the national education system Detailed description of how STEM education is contemplated and/or approached in the country's educational system and the different regulations, guidelines, calls, programs, existing initiatives in this regard. Including references to the corresponding documentation that should be included as a bibliography. Detailed description of how STEM education is contemplated and/or approached in the country's educational system and the different regulations, guidelines, calls, programs, existing initiatives in this regard. Including references to the corresponding documentation that should be included as a bibliography. Detailed description of how STEM education is contemplated and/or approached in the country's educational system and the different regulations, guidelines, calls, programs, existing initiatives in this regard. Including references to the corresponding documentation that should be included as a bibliography. Digital assessment in the national education system 104 Detailed description of how digital assessment is contemplated and/or approached in the country's educational system and the different regulations, guidelines, calls, programs, existing initiatives in this regard. Including references to the corresponding documentation that should be included as a bibliography. 2.2. DATA ANALYSIS How the analyses of data from the different instruments have been carried out and how their triangulation has been ensured in the preparation of the report. 1.2. CONTEXT OF THE PILOT SCHOOLS This section should include the following information on the overall description of the schools participating in the pilot Number of centers and their distribution according to: for example, context (rural, urban, semi-urban), ownership (public/private), type of center... Ownership (public, private, arranged) Composition of the school: number of students, number of teachers Defining features (pedagogical projects) that can define the specific identity for each of the schools, if it has any (for example a commitment to digitalisation, sustainability, interculturality, diversity...) 1.3.2. DETAILED DESCRIPTION OF THEATS-STEM PROJECTS DEVELOPED DURING THE PILOT Annexing the designs of the learning cycle (WP3) and illustrating it with some images of the implementation process CHAPTER 2. METHODOLOGY Annexing the designs of the learning cycle (WP3) and illustrating it with some images of the implementation process CHAPTER 2. METHODOLOGY 2.1. DATA COLLECTION Annexing the designs of the learning cycle (WP3) and illustrating it with some images of the implementation process CHAPTER 2. METHODOLOGY 2.1. DATA COLLECTION 105 In this section we must collect information about: 2.1.1. Process followed for data collection through the instruments Electronic questionnaires on student core ATS-STEM learning competences: number and process of application Classroom observations: number and process of application Observations in the virtual environment: number and process of implementation Observations in the virtual environment: number and process of implementation Selection of devices/products to be analysed from the student body: number and process of applicatio Selection of devices/products to be analysed from the student body: number and process of application Conducting interviews with the different agents involved in the ATS-STEM projects: teachers, students, and mentors: number and process of application Conducting interviews with the different agents involved in the ATS-STEM projects: teachers, students, and mentors: number and process of application 2.1.2. Incidents to be taken into account in data collection: relating any incidents that may have occurred. In addition, information on deviations from data collection in relation to planning/forecasting should be incorporated. 2.1.2. Incidents to be taken into account in data collection: relating any incidents that may have occurred. In addition, information on deviations from data collection in relation to planning/forecasting should be incorporated. 2.3. QUALITY GUARANTEE In this section, information must be provided on how the data has been processed, both to guarantee the confidentiality of the data as well as to ensure that the data is processed rigorously in terms of its validity and reliability. (a) Extraction of data from different sources of information, (b) stability technique: the same observer observes at different times (Bisquerra, 1989). Authorisation by ethics committees if appropriate and annexed 3.1. RESULTS DESCRIPTION Organised around the following research questions and including references from the pilot (literal quotations from the interviews, digital interactions captured from the DA environment, critical incident photos, etc) that allow each question to be exemplified from the observation sheets (both in the classroom and in the virtual environment) as well as from the analysis of artefacts and interviews carried out. 3.1.1. Digital assessment description 3.1.1.1. What digital assessment tools have teachers and students used? And how have they been used? (proposal for use by teachers within the framework of ATS-STEM projects) 3.1.2. Formative assessment achievements 3.1.2.1. Integrate ATS-STEM content (and learning outcomes/goals) Addressing core ATS-STEM learning outcomes/objectives/competences 106 Interdisciplinarity and globalised approach 3.1.2.2. Reflect ATS-STEM learning design principles (and social constructivist views of learning) Working in the classroom with problems and challenges Reflecting social constructivist views on learning Stimulating creativity to offer more than one solution to problems 3.1.2.3. Facilitate feedback (improve learning by prompting the learner to use effective feedback focused on the learning outcome/goal in a timely manner Feedback about the process Feedback related to self-regulation 3.1.2.4. Facilitate peer-assessment (improve learning by activating students as instructional resources for one another) 3.1.2.5. Facilitate self-assessment (improve learning by activating students as owners of their own learning) 3.1.2.6. Help to Elicit evidence of learning (improve learning through questioning and discussion and by prompting activities that clarify the meaning of success) 3.1.3. Challenges in digital assessment 3.1.3.1. What are the main difficulties? 3.1.3.2. Proposals and recommendations from the agents involved CHAPTER 4. CONCLUSIONS AND PROPOSALS In this chapter, the main conclusions derived from the data discussed above will be formulated, trying to relate them to the previous scientific literature: Conclusions following the assessment How might digital assessment practices support the development of STEM competencies? How and with which digital assessment methodologies and tools can we improve ATS-STEM teaching and learning processes? Why apply digital assessment in the development of ATS-STEM projects? What and how does it contribute for STEM teaching and learning processes? What are the challenges to using digital assessment strategies in ATS-STEM learning? Limitations of the study Recommendations and proposals for the implementation of ATS-STEM and digital assessment projects in schools. 3.1. RESULTS DESCRIPTION For teachers and students In this chapter, the main conclusions derived from the data discussed above will be formulated, trying to relate them to the previous scientific literature: Conclusions following the assessment How might digital assessment practices support the development of STEM competencies? How and with which digital assessment methodologies and tools can we improve ATS-STEM teaching and learning processes? How might digital assessment practices support the development of STEM competencies? How and with which digital assessment methodologies and tools can we improve ATS-STEM teaching and learning processes? Why apply digital assessment in the development of ATS-STEM projects? What and how does it contribute for STEM teaching and learning processes? Why apply digital assessment in the development of ATS-STEM projects? What and how does it contribute for STEM teaching and learning processes? What are the challenges to using digital assessment strategies in ATS-STEM learning? Recommendations and proposals for the implementation of ATS-STEM and digital assessment projects in schools. For teachers and students Recommendations and proposals for the implementation of ATS-STEM and digital assessment projects in schools. For teachers and students • To be taken into account in educational policies 107 107 5. REFERENCES Aguilar, S., & Barroso, J. (2015). La triangulación de datos como estrategia en investigación educativa. Píxel-Bit. Revista de Medios y Educación, 47, 2171-7966. doi: http://dx.doi.org/10.12795/pixelbit.2015.i47.05 Betrián Villas, E., Galitó Gispert, N., García Merino, N., Jové Monclús, G., & Macarulla Garcia, M. (2013). La triangulación múltiple como estrategia metodológica. REICE. Revista Iberoamericana sobre Calidad, Eficacia y Cambio en Educación, 11(4), 5-24. Retrieved from https://www.redalyc.org/pdf/551/55128238001.pdf Betzner, A., Lawrenz, F. P. & Thao. M. (2016). Examining mixing methods in an assessment of a smoking cessation program. Assessment and Program Planning, 54, 94-101. DOI: http://dx.doi.org/10.1016/j.evalprogplan.2015.06.004 Correa, S., Puerta, A., y Restrepo, B. (2002). Investigación evaluativa. Bogotá, Colombia: ICFES Christie, C. A. & Fleischer, D. N. (2010). Insight Into Assessment Practice: A Content Analysis of Designs and Methods Used in Assessment Studies Published in North American Assessment-Focused Journals. American Journal of Assessment, 31(3), 326-346. DOI: http://dx.doi.org/10.1177/1098214010369170 Escudero, T. (2016). La investigación evaluativa en el Siglo XXI: Un instrumento para el desarrollo educativo y social cada vez más relevante. RELIEVE, 22 (1), art. 4. DOI: http://dx.doi.org/10.7203/relieve.22.1.8164 Expósito, J. (2003). Análisis cientimétrico, conceptual y metodológico de la investigación española sobre evaluación de programas educativos (1975/2000). (Tesis doctoral). Recuperado de https://dialnet.unirioja.es/servlet/tesis?codigo=59487 Exposito, J., Olmedo, E. & Fernandez-Cano, A. (2004). Patrones metodológicos en la investigación española sobre evaluación de programas educativos. RELIEVE, 10 (2). DOI: http://dx.doi.org/10.7203/relieve.10.2.4319 Exposito, J., Olmedo, E. & Fernandez Cano, A. (2004). Patrones metodológicos en la investigación española sobre evaluación de programas educativos. RELIEVE, 10 (2). DOI: http://dx.doi.org/10.7203/relieve.10.2.4319 Makrakis, V. & Kostoulas-Makrakis, N. (2016). Bridging the qualitative–quantitative divide: Experiences from conducting a mixed methods assessment in the RUCAS programme. Assessment and Program Planning 54, 144-151. DOI: http://dx.doi.org/10.1016/j.evalprogplan.2015.07.008 Perassi, Z. (2009). Commentary: Evaluar un Programa Educativo: Una Experiencia Formativa Compleja. Revista Iberoamericana de Evaluación Educativa, 2(2), 172-195. 09). Commentary: Evaluar un Programa Educativo: Una Experiencia Formativa Compleja. Revista mericana de Evaluación Educativa, 2(2), 172-195. Suchman, E. (1967). Evaluative research. Nueva York: Russell Sage Foundation. Suchman, E. (1967). Evaluative research. Nueva York: Russell Sage Foundation. Tejedor, F. J., García-Varcárcel, A., y Rodríguez Conde, Mª. J. (1994). Perspectivas metodológicas actuales de la evaluación de programas en el ámbito educativo. Revista Investigación Educativa, 23, 93-127. White, H. (2013). The Use of Mixed Methods in Randomized Control Trials. New Directions for Assessment, 138(2), 61-73. DOI: http://dx.doi.org/10.1002/ev.20058 108 108	24,430
52/tel.archives-ouvertes.fr-tel-00676796-document.txt_10	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	9,954	15,566	singulariser l'événement. Le batteur prend alors le relais sur ses toms (très discrètement à partir de 7'52'', puis, le reste de la musique s'étant éteint, plus en dehors à 8'04''). 16) La musique va rester sur les deux caractéristiques principales de cette proposition initiale : à la frontière du silence ; et très bruiteux (le retour des ocarinas à 0'44'', piste 5). Du reste, c'est encore une fois un solo (ici de batterie, uniquement sur les toms) qui sert à créer l'articulation entre deux épisodes. Les autres musiciens se greffent simplement dessus pour « déclarer » le début d'une nouvelle séquence collective. Un chant de saxophone baryton, très fragile, émerge d'abord. Remarquons encore une fois le marqueur formel qui précède l'entrée de la clarinette (à 2'51'') : la basse vient doubler le mi du saxophone à 2'46'', créant ainsi un signe d'articulation claire. Toutefois, la basse quitte aussitôt le domaine des hauteurs (revenant à un jeu percussif très en arrière-plan), pour laisser s'épanouir un duo lyrique saxophone-clarinette. Son intervention a donc une valeur strictement formelle (créer une articulation en singularisant, par le redoublement d'une hauteur, un événement), indépendante de toute motivation musicale (puisque son intervention n'est absolument pas exploitée par la suite). 17) La musique se vide de plus en plus de toute intention thématique, pour se transformer en mélodie de timbre très espacée. Il est alors clair pour tous que la musique est en train de s'éteindre, et qu'il s'agit là de la fin. L'apparition d'une boucle bruiteuse (réalisée par le bassiste sur petite boîte d'effets) à 5'24'' va permettre de conclure l'improvisation. Cette boucle est d'abord comme une simple texture, environnant un dernier sursaut lyrique autour de quelques notes (avec le retour du piano, dans l'extrême aigu). Mais après un dernier mi de piano (sorte de finale de ces bribes mélodiques qui viennent de s'effacer), cette texture passe au premier plan (d'autant plus que le bassiste en augmente progressivement le volume). Tout le monde comprend qu'il ne va plus rien se passer désormais, et, effectivement, rien ne se passe (en dehors de quelques imitations bruiteuses à la clarinette, qui se fondent dans la texture). Le retour progressif du son d'arpégiateur inaugural (à 6'46'') permet de créer une véritable coda, et d'éviter la traditionnelle fin des improvisations collectives libres où la musique n'en finit plus de mourir et de retourner à la pure trame. 4. Transitions, marqueurs formels et articulations entre les séquences Cette narration détaillée ayant atteint son terme, essayons maintenant de distinguer les différentes manières dont s'opèrent les articulations formelles au sein de cette improvisation. Nous avons vu que cette improvisation pouvait assez facilement être divisée en diverses séquences, relativement autonomes et caractérisées, reliées linéairement (de proche en proche : on retrouve donc notre idée de forme concaténée). Il s'agit maintenant de comprendre comment s'effectue le passage d'une séquence de l'improvisation à une autre. On comprend évidemment l'importance de ce point puisque nous avons relevé dans la partie précédente la nature profondément séquentielle de la forme improvisée. À notre sens, une séquence contient deux « ingrédients » essentiels (c'est-à-dire définitionnels) : 200 1) un certain schéma d'interaction c'est-à-dire la manière dont les interactions se sont cristallisées, en moyenne, sur une durée longue : typiquement, qui envoie beaucoup d'information, qui en envoie peu, et la constitution de sous-équipes (qui est en interaction privilégiée avec qui) ; 2) et un ensemble de caractérisations acoustiques, cinétiques et/ou musicales valant, en moyenne, pour la durée considérée. On peut considérer qu'il y a changement de séquence à chaque fois que le schéma d'interaction change ; ou à chaque fois que la caractérisation change ; ou quand les deux ont changé. Une manière de simplifier le problème (et pas forcément fausse) est de dire que, d'une manière générale, les deux ingrédients tendent à changer en même temps. C'est-à-dire qu'un changement dans le schéma d'interaction aura tendance à forcer les improvisateurs à redéfinir la caractérisation de la séquence, et ciproquement. La re-coordination qui s'effectue à chaque nouvelle séquence porterait donc sur les deux ingrédients à la fois : il s'agirait alors à la fois de trouver une nouvelle caractérisation commune, et le schéma d'interaction qui permet de l'exprimer de manière privilégiée. On remarque en fait que, dans l'improvisation en question, c'est presque toujours une modification du schéma d'interaction qui est à l'origine du changement de séquence. En suivant Nunn (1998), on peut distinguer sept manières d'articulation entre les séquences : 1) Transition soudaine et inattendue : il n'y a pas d'effet de clôture de la séquence en cours. Une nouvelle séquence, en général contrastante, s'enchaîne directement, sans pause ni interruption. 5) Transition par modification d'un trait caractéristique : la transition est en général relativement longue car celle-ci consiste en la modification graduelle et progressive d'un des éléments précédents, permettant une redirection du flux musical. Cette transition ne produit pas d'effet de clôture. 6) Transition par fragmentation : une certaine texture, ou un certain rythme, se trouve progressivement brisé, fragmenté, entrecoupé de silence. Cette transition produit un effet de clôture. 7) Cadence interne : des gestes cadentiels conduisent la séquence à son terme, jusqu'au silence, suivi par une nouvelle séquence. Il y a bien sûr un effet de clôture. Il faut noter qu'il y a une grande imprévisibilité dans ces moments, parce qu'il n'est jamais certain que l'improvisation reparte. Autrement dit, ce genre de cadence prend toujours le risque de conduire à la fin de l'improvisation. Ce découpage est évidemment très intéressant, et pourrait sans doute constituer un bon point de départ pour l'analyse des transitions, d'autant plus que Thomas Nunn suggère qu'on peut aussi imaginer des transitions hybrides, c'est-à-dire qui combinent différents aspects des catégories pures énumérées ci-dessus. Toutefois, cette typologie nous semble se heurter à deux problèmes : 1) D'abord, il est difficile de rencontrer ces catégories à l'état « pur », surtout dans le cas de l'improvisation collective libre, où des processus formels différents selon les musiciens ont tendance à se superposer. Regardons par exemple la toute première transition de notre improvisation : comment la caractériser selon le découpage de Nunn? Le bassiste opère une prise de parole plutôt soudaine et inattendue ; le saxophoniste et le clarinettiste sont quant à eux plutôt en train de se cristalliser sur un motif directement issu de la séquence précédente ; quant au synthétiseur, il opère un ralenti très progressif du débit qui dominait la séquence précédente. C'est donc bien d'un hybride entre trois types de transition qu'il s'agit. 2) Ensuite, certaines de ces transitions s'étendent sur des durées non négligeables, de sorte qu'elles peuvent très bien acquérir le statut de séquence. Il nous semble plus raisonnable de ramener le catalogue de Nunn, de toute façon insuffisant, puisqu'il faut sans cesse imaginer des cas intermédiaires ou hybrides, à trois grandes catégories d'articulation : 202 1) La juxtaposition : une séquence est arrivée à son terme « naturel » (on observe un ou plusieurs mouvements cadentiels), par le « haut » (transition par climax) ou par le « bas » (résolution, cadence interne), les improvisateurs sont donc en attente d'une nouvelle séquence. La première proposition à surgir a donc toutes les chances de caractériser en bonne partie la séquence à venir. 2) L'interruption : une séquence est en cours mais elle se trouve brusquement perturbée par un ou plusieurs improvisateurs qui introduisent a priori des éléments très contrastants, susceptibles de jouer efficacement un rôle d'interruption. La séquence peut ou bien être effectivement interrompue, pour laisser la place à une séquence dominée par ces éléments contrastants, ou bien continuer malgré tout, et se trouver donc superposée aux éléments contrastants. 3) L'interpolation : la séquence suivante ne s'établit que très progressivement, éventuellement à partir des éléments de la séquence précédente. Cela désigne en fait tout simplement le processus général qui est à l'oeuvre dans certaines transitions dont Nunn détaille les modalités (fragmentation, transition par modification d'un trait caractéristique, transition par trait caractéristique commun). Ainsi, selon ce découpage, nous entendons plutôt la première transition comme une tentative d'interruption (l'introduction des éléments pastoriussiens à la basse) sans effet immédiat, qui se retrouve donc superposée aux reliquats de la séquence précédente (les motifs rythmiques entrecroisés du saxophone et de la clarinette). Ce découpage permet d'ailleurs de comprendre une ambiguïté notre usage du terme de « transition ». En effet, nous l'utilisons pour désigner, assez naturellement, le passage d'une séquence à une autre. Mais le fait même que ces transitions puissent être de durée fort variable peut nous amener, parfois, à vouloir ériger la transition elle-même en séquence. C'est typiquement le cas quand nous avons affaire à une interpolation ou à une interruption « avortée ». C'est pour cela que nous n'avons pas hésité, au début de cette analyse, à diviser nos séquences en épisodes et transitions. Les premiers sont des séquences caractérisées par le maintien ou le développement d'une identité ; les secondes sont des séquences processuelles, où une identité se désagrège pour laisser place à une autre. Ainsi, la partie qui s'étend de 3'20'' à 5'38'' (piste 1) est bien une séquence ; mais en tant qu'on y trouve de nombreux éléments conflictuels, et donc en raison de son caractère éminemment instable, on peut aussi considérer cette séquence tout entière comme une transition. 203 C'est pourquoi nous ne nous intéresserons pas spécialement dans nos analyses aux transitions mais bien plutôt aux articulations formelles entre les différentes séquences, fussent-elles des transitions. À cet égard, l'élément le plus remarquable mis en avant par ces analyses est sans doute la présence systématique de ce que nous avons appelé les marqueurs formels. Nous les caractérisons comme « formels » parce qu'ils sont, en un sens, vides de contenu : ils ne disent le plus souvent rien sur la nature musicale du discours improvisé. En revanche, on peut observer que les improvisateurs utilisent sans cesse des éléments saillants par rapport au contexte, comme prétexte au passage d'une séquence à une autre. Ces marqueurs formels peuvent être de différents types : 1) un élément différencié, d'une manière ou d'une autre, à l'intérieur d'un faisceau d'événements (un son à hauteur perceptible au sein d'un discours bruiteux) ; 2) un élément de type « accidentel », que ce soit au niveau individuel (un son mal maîtrisé) ou au niveau collectif (la simultanéité de deux attaques dans une musique non pulsée) ; 3) un événement disposant d'une saillance immédiate (un son très fort et brusque, par exemple) ; 4) une articulation (contraste ou interruption) produite unilatéralement par un improvisateur (l'entrée d'un improvisateur qui ne jouait pas jusqu'à présent). Les musiciens pratiquant l'improvisation libre ont, de manière générale, une sensibilité à la exacerbée : cela s'observe notamment dans l'effort d'ajustement quasi-permanent des faisceaux à l'oeuvre dans l'improvisation collective libre, qui est déjà un premier moyen de résoudre le problème de coordination temporelle. Mais certains événements acquièrent, dans un contexte donné, suffisamment de saillance (les fameux marqueurs formels) pour que les musiciens les utilisent pour opérer collectivement une articulation formelle. On comprend alors l'importance de ces événements dans le cadre qui est le nôtre, où la forme est avant tout envisagée comme séquentielle, comme une concaténation de moments liés les uns aux autres. Et il est probable que ce soit ces marqueurs formels qui fassent typiquement l'objet d'une procédure de raisonnement d'équipe dans l'improvisation collective libre. Notons que cela ne résout en rien le problème du contenu musical : sur quoi la coordination des musiciens doitelle s'effectuer? L'hypothèse que nous proposons, c'est qu'il existe des phénomènes de contagion entre marqueurs formels et points focaux ; c'est-à-dire que la présence d'un 204 marqueur formel va singulariser une proposition musicale, la constituant en point focal pour le groupe d'improvisateurs (ce dont témoigne par exemple la rapidité avec laquelle les musiciens vont se regrouper autour de cette proposition). Dans l'improvisation qui nous occupe, on peut constater différentes manières d'opérer l'articulation séquentielle à partir de ces marqueurs formels (manières non exclusives les unes des autres) : 1) Le marqueur formel amène tous les musiciens à opérer des changements graduels, typiquement en cascade : passage de 1 à 2, de 2 à 3, de 15 à 16 2) Le marqueur formel provoque un changement d'orchestration (retrait et/ou apparition d'un ou plusieurs improvisateurs), ces changements d'orchestration s'étageant souvent en cascade : passage de 3 à 4, de 5 à 6, de 7 à 8, de 8 à 9, de 16 à 17 3) Plusieurs marqueurs formels sont articulés entre eux, produisant par là un effet de confirmation intersubjective : passage de 2 à 3, de 6 à 7 4) Variante du cas précédent : le marqueur formel crée une identité mais c'est la rupture de cette identité qui va devenir saillante et fonctionner à son tour comme marqueur formel : passage de 10 à 11 5) Le marqueur formel est lui-même l'élément saillant de coordination. C'est souvent le cas quand le marqueur formel est le fait d'un changement d'orchestration, ou d'une articulation autonome de la part d'un des improvisateurs : passage de 2 à 3 6) Le marqueur formel permet de singulariser une proposition par un effet de contagion : passage de 5 à 6, de 6 à 7, de 8 à 9, de 13 à 14 Ces événements sont essentiels pour comprendre la dynamique d'une improvisation collective libre. En effet, les musiciens font sans doute le pari d'une certaine transparence de la saillance qui caractérise en propre le marqueur formel ; elle est la seule à pouvoir prétendre devenir une connaissance commune (tout le monde a remarqué tel événement, et tout le monde sait que tout le monde l'a remarqué). B/ Les Company Weeks Le guitariste Derek Bailey, pionnier de l'improvisation libre « non-idiomatique » comme nous l'avons déjà vu, a été le fondateur d'un festival d'improvisation, à la fréquence quasiannuelle entre 1977 et 1994. Il s'agissait d'un festival de musiques improvisées au principe original (même si on peut trouver aujourd'hui quelques festivals reposant sur un principe analogue, comme le festival de Moers) : constituer un ensemble de musiciens, de pays différents, qui pour la plupart n'ont jamais joué ensemble, et les faire improviser selon différentes combinaisons (du solo au tutti) lors de nombreux concerts répartis sur quatre ou cinq jours. Derek Bailey explique très clairement la motivation sous-jacente à ce principe : « Depuis quelque temps, il me semble que l'improvisation libre la plus intéressante est le fait de formations relativement éphémères []. Dans ce genre de musique, j'avais toujours trouvé que les débuts d'un groupe constituaient la période la plus satisfaisante, la plus stimulante. Lorsqu'une musique acquiert une identité assez forte pour être analysée, décrite et bien sûr reproduite, tout change. Parvenu à une certaine maturité et ayant découvert une musique à laquelle il s'identifie, le groupe atteint un point où, bien qu'il puisse continuer à se développer sur le plan musical et devenir plus commercial - attractions souvent irrésistibles - la musique cesse d'être de l'improvisation pure » (Bailey 1999, p. 143). Cette volonté de chercher l'improvisation la plus « pure », la plus radicale, fait tout le prix de la démarche de Derek Bailey. C'est aussi pourquoi les quelques enregistrements existants de Company sont pour nous des objets d'analyse privilégiés. Non qu'il faille faire mentir Derek Bailey, quand il dit que la musique des débuts d'un groupe d'improvisation est encore inanalysable. Pour nous, il est certain que l'analyse ne porte pas tant sur l'enregistrement en tant qu'objet fini, servant à dégager les traits caractéristiques d'une idiosyncrasie, mais sur l'enregistrement comme témoignage d'un processus, capturant au plus près ce qui fait la radicalité de l'improvisation collective libre, et permettant dès lors de poser la question du « comment? ». Comment ces gens qui ne se connaissent pas et ne partent pas d'un référent commun parviennent-ils à faire de la musique ensemble? Et d'ailleurs, la question du succès ou de l'échec de cette entreprise en devient presque secondaire (et l'on sait à quel point cette musique librement improvisée est fragile, et que les réussites éclatantes y sont peu nombreuses) : ce qui importe, c'est de pouvoir comprendre les mécanismes cognitifs sous- 206 jacents, les stratégies que mettent en place les improvisateurs pour essayer de créer collectivement de la musique. D'ailleurs, c'est bien souvent ce que recherchent les amateurs de musiques improvisées lorsqu'ils assistent à un concert : pas forcément du sublime, mais de l'intelligent ; des musiciens qui essayent collectivement de se sortir d'un problème particulièrement ardu dans lequel ils viennent de s'enfoncer. L'improvisation collective libre est bien une situation de type « problem-finding » et non de type « problem-solving » pour reprendre la distinction faite par K eith Sawyer (2003)250 : ce qui est intéressant, c'est de trouver de nouveaux problèmes. Par exemple, le problème n'est pas de savoir « comment les improvisateurs en viennent à partager des représentations identiques mais comment une interaction cohérente peut se produire quand ils ne partagent pas ces représentations251 » (Sawyer 2003, p. 9, c'est nous qui soulignons). Il y a beaucoup de points communs entre le plaisir que l'on a à écouter de l'improvisation collective libre et le plaisir d'écouter une véritable conversation : dans les deux cas, il s'agit du plaisir d'écouter les gens penser publiquement (la finesse dans l'enchaînement des arguments, la drôlerie d'une répartie, la négociation d'un changement de sujet). L'idée de forum d'improvisation prend ici tout son poids : l'improvisation libre apparaît comme un lieu de rencontre où des personnes qui ont en commun l'improvisation peuvent se retrouver. Évidemment, cette idée est éminemment discutable : en quoi le partage d'une modalité de discours, d'expression, d'une manière particulière de produire du musical (l'improvisation), pourrait-il être garant de la possibilité d'une véritable communication musicale? Pour le dire plus crûment, est-ce parce que nous avons une manière commune de dire les choses que nous pouvons avoir des choses communes à dire? Pour Derek Bailey, là n'est pas vraiment la question : encore une fois, l'idée sous-jacente à ce mélange des familles d'improvisateurs est de garantir la radicalité de l'improvisation libre : non seulement les musiciens ne se connaissent-ils pas, mais encore ne pourront-ils pas compter sur des codes et conventions implicites d'improvisation inhérents à leur milieu musical. Il s'agit donc de créer la situation la plus difficile et ingrate a priori, pour observer dans toute sa créativité le processus d'improvisation collective libre ; et encore une fois, ce n'est pas tant le résultat qui intéresse Derek Bailey, encore que celui-ci se trouve assez souvent satisfait de la musique ainsi produite, que le processus. Ceci est clairement revendiqué par Derek Bailey quand, par exemple, il décide d'inviter des non-improvisateurs (comprendre : des musiciens classiques, par exemple le corniste du London Sinfonietta) pour être certain qu'aucun des musiciens ne pourra se reposer sur un métier partagé d'improvisateur. « Durant les premières années de Company, les musiciens que j'invitais étaient toujours choisis parmi des gens surtout passionnés par l'improvisation. J'ai cependant tenté de rassembler des musiciens pour qui l'improvisation avait des fonctions différentes et qui, dans de nombreux cas, ne se connaissaient pas. À mon avis, cela fonctionna assez bien, mais en 1982, la situation était devenue un peu trop prévisible. Peut-être en raison d'une stagnation musicale générale, alors que dans presque tous les domaines (comme aujourd'hui d'ailleurs) on notait un enthousiasme croissant pour l'imprévisibilité totale. Ou peut-être jouer avec absolument n'importe quel improvisateur était-il devenu si courant pour les musiciens que les différences avaient perdu toute importance. Il s'agit donc bien de rendre à l'improvisation son étymologie, de faire d'elle, purement et radicalement, une situation non seulement imprévue (ce qui est le propre de l'improvisation) mais imprévisible (ce qui devrait être le propre de l'improvisation collective libre, qui, si ces deux réquisits sont pris au sérieux, devrait être une situation à la dynamique hautement instable). Cette idée (introduire des non-improvisateurs au sein d'improvisateurs) est fructueuse en raison d'une asymétrie fondamentale : en effet, non seulement, cela empêche l'improvisateur 208 de se reposer « sur des pratiques généralement considérées comme sûres » (ibid., p. 146) mais en plus cette radicalisation ne se fait pas au prix du ridicule du non-improvisateur qui, d'après Bailey, « discernera les divers repères musicaux et les intentions (pratique sensorielle courante) sans difficultés majeures » (ibid., p. 146). Au fond, c'est exactement la même idée qui rend séduisant le forum d'improvisation : ce n'est pas tant le vocabulaire utilisé (l'idiome employé, pour reprendre le vocabulaire de Bailey) qui fait la qualité d'un improvisateur, que cette « pratique sensorielle courante », partagée par la plupart des musiciens de haut niveau, improvisateurs ou non : la capacité de saisir des intentions musicales à travers des signaux sonores. C'est cette oreille du musicien qui est particulièrement sollicitée dans le processus créateur inhérent aux rencontres d'improvisation libre. C'est pour cette raison que la musique librement et collectivement improvisée peut difficilement être idiomatique. Le but étant la création collective d'une musique par l'improvisation, et les improvisateurs venant d'horizons musicaux assez variés, il faut, pour que l'idée de création collective ne soit pas vide, que chacun puise en-deçà/au-delà des idiomes qu'il maîtrise par ailleurs. Ben Watson (2004)252 souligne cette difficulté en constatant que les deux passages les plus faibles de Once (enregistré lors du Company Week de 1987) sont précisément les passages où certains improvisateurs (en l'occurrence Lee Konitz et Carlos Zingaro) se laissent aller directement à une évocation trop précise d'un idiome déjà établi (respectivement le jazz, avec une citation de As Time Goes By, et la tradition du violon folk) : « Bien sûr, les musiciens qui improvisent doivent bien venir de quelque part []. C'est juste que « l'identité musico-sonore » du jeu des individus doit être consciente du son d'ensemble, et non pas reposer sur quelque plan linéaire et préconçu ou sur des citations d'un savoir musical positif253 » (Watson 2004, p. 255). Derek Bailey est très clair sur l'objectif de son festival : « Le but de Company est de créer collectivement de la musique. Cela peut exiger le sacrifice de ses préférences individuelles ainsi que de la générosité, de la curiosité, de la sensibilité et une aptitude à réagir instinctivement et de façon constructive à des situations imprévues » (Bailey 1999, p. 147). « Of course, musicians who improvise must come from somewhere []. It's simply that the « sonic-musical identity » of the individual's playing needs to be cognisant of the ensemble sound, and not off on some preconceived linear trop, some citation of positive musical culture ». Cette idée est très importante pour notre propos : en effet, un des grands paris de notre approche est l'existence dans l'improvisation collective libre de préférences collectives, qui coexistent avec les préférences individuelles des musiciens, sans s'y réduire ni les écraser. Certes, il y a plusieurs niveaux d'interprétation de cette articulation préférences collectives/préférences individuelles, mais à un niveau assez global, on peut la relier de manière pertinente avec l'articulation improvisation idiomatique (entendu au sens très large)/improvisation non-idiomatique. Chaque improvisateur vient d'une certaine tradition musicale, avec ses schémas de pensée musicale bien établis et un certain vocabulaire, fait d'innovations et de clichés Pourtant, lorsque vient le moment de l'improvisation collective libre, ces préférences d'improvisation peuvent (voire doivent) laisser la place à des préférences collectives construites sur le moment et découlant d'une agrégation de musiciens particulière. À notre sens, on peut interpréter une improvisation réussie comme une improvisation où les musiciens ont réussi à constituer des préférences collectives, et donc à prendre au sérieux l'idée d'improvisation non-idiomatique. Ou pour le dire autrement, c'est parce que les musiciens parviennent à cadrer la situation d'improvisation collective libre comme problème de coordination que des préférences collectives apparaissent. Néanmoins, nous l'avons déjà dit, Derek Bailey est le premier à nuancer cette opposition entre improvisation idiomatique et improvisation non-idiomatique : « Il me semble aujourd'hui que, dans la pratique, la différence entre l'improvisation libre et l'improvisation idiomatique n'est pas fondamentale. Pour l'improvisateur libre, la liberté est comme le summum de l'expression idiomatique, pour l'improvisateur idiomatique, une sorte d'utopie. Dans la pratique, les deux genres de musiciens se concentrent probablement plus sur les moyens que sur le but à atteindre. Toute improvisation a lieu en relation avec ce qui est connu, qu'il s'agisse d'éléments traditionnels ou nouvellement acquis. La seule véritable différence réside dans les occasions qu'a l'improvisation libre de renouveler ou de modifier l'inconnu et d'ouvrir ainsi une perspective illimitée inexistante, par définition, dans l'improvisation idiomatique » (Bailey 1999, p. 152, c'est nous qui soulignons). Dans les deux cas, la question centrale est donc plutôt celle du « comment? », plutôt que celle du résultat ; et ce « comment? » est à comprendre comme une certaine manière d'articuler les éléments connus et les éléments inconnus. L'improvisation collective libre a ceci d'extraordinaire qu'elle oblige les musiciens à se poser cette question de l'articulation du connu et de l'inconnu avec une radicalité et une acuité uniques. Et toute la tâche de Derek Bailey dans l'organisation de ces Company Weeks a été de veiller à ce que cette question ne cède pas la place à une réponse définitive. Méthodologie et présentation des analyses Chaque analyse est accompagnée d'une partition de lecture, à laquelle il convient de se reporter : ces « partitions » sont présentes ici même, au début de chaque analyse. Mais on les trouvera également au format PDF dans le CD-Rom joint, pour plus de confort de lecture254. Ce CD-Rom comporte également les enregistrements des improvisations considérées (au format MP3). Nous n'avons d'aucune façon tenté de noter la musique analysée : outre la complexité folle d'une telle entreprise (la musique est essentiellement non-pulsée et les musiciens utilisent une foule de techniques instrumentales qui ne se laissent pas vraiment capturer par une notation traditionnelle), sa pertinence ne semble pas acquise. Même à supposer que nous parvenions à trouver des outils de notation appropriés, nous ne voyons pas pourquoi le passage par l'écrit du contenu pourrait nous aider à mieux comprendre les mécanismes de coordination à l'oeuvre dans cette musique. En revanche, il nous a semblé essentiel de proposer quelque chose qui permettait de visualiser d'un seul coup d'oeil les interactions et la structure de l'improvisation. Nous avons conservé un mode de représentation absolument traditionnel : la partition de lecture est organisée selon deux axes : le temps s'écoule, en secondes, sur l'axe horizontal (de gauche à droite) ; les différents instruments sont indiqués sur l'axe vertical. C'est essentiellement d'une représentation formelle qu'il s'agit. Nous avons essayé d'indiquer les faisceaux d'événements de chaque improvisateur, selon la formalisation qui a été proposée à la fin de la partie précédente. Ne pouvant entrer dans la tête des improvisateurs, c'est bien évidemment à une reconstruction que l'on a affaire. Mais il vient de noter que c'est également ce genre de reconstruction qui a cours à l'intérieur même d'une improvisation, entre musiciens ; la différence majeure étant bien sûr que nous disposons du privilège du temps différé et de la réécoute, privilège exorbitant pour l'improvisateur qui, lui, est soumis à l'exigence d'une reconstruction cognitive en temps réel. La démarche est donc essentiellement inductive (mais, encore une fois, c'est également de cette manière que procèdent les participants d'une même improvisation) ; dès qu'un changement notable (plus ou moins notable d'ailleurs) se fait sentir dans le discours d'un des improvisateurs, on peut raisonnablement supposer qu'il y a eu prise de décision. changement de processus (c'est-à-dire une modification dans la manipulation de ces matériaux). Nous avons systématiquement indiqué les silences (dès qu'un improvisateur ne joue plus, le faisceau est d'autorité achevé), car cela permet de comprendre certains mécanismes de coordination de base : en effet, commencer une proposition pendant que l'autre se tait, c'est minimiser les possibilités de discoordination entre les différents discours. Les faisceaux sont de taille extrêmement variable. Ce n'est pas un problème en soi, mais dans certains cas, nous franchissons allègrement les limites du réalisme cognitif : un faisceau (qui, rappelons-le, rassemble en une unité cognitive un ensemble d'événements décidés en un même point) ne peut raisonnablement excéder 10 secondes en situation d'improvisation collective. Pourtant, certains de nos faisceaux franchissent cette limite : cela signifie que nous n'avons pas perçu de modification significative, ou disons pertinente dans le contexte, et que nous avons considéré que tous les événements ressortissaient du même faisceau. En fait, il y a très certainement sous ce genre de faisceau une multitude de faisceaux générés par association par similitude. Mais, encore une fois, il y a des chances que les co-improvisateurs se livrent au même genre d'interprétation Ce genre de représentation, extrêmement formelle, permet de voir rapidement le jeu des interactions dans la musique qui nous occupe. Nous indiquons parfois, quand cela est pertinent pour l'analyse, les rapports qui existent entre les différents discours (imitation, contraste) ; apparaissent également (comme indiqué dans la légende ci-après) quelques éléments du contenu musical des discours, notamment tout ce qui permet de faciliter la coordination des discours (polarisation, tenue ou répé , introduction d'une pulsation), que ceux-ci résultent d'une tentative de coordination explicite ou de l'utilisation d'une stratégie d'attente (notamment dans le cas de la répétition) : dans les deux cas, ces stratégies créent un pôle de stabilité (maintien d'une identité) et permettent aux co-improvisateurs la mise en place de modèles prédictifs efficaces. Finalement, nous avons porté dans notre schéma ce qui pouvait être particulièrement saillant pour les improvisateurs : les prises de décision de type « articulatoire » (que ce soit donc, dans les termes de Pressing, une génération par association par contraste ou une génération par interruption) ou les éléments de type « accidentel » (qu'il s'agisse d'un événement vraiment accidentel, ou bien d'un élément de discours particulièrement surprenant ou contrastant à l'intérieur de ce qui semble être un unique faisceau d'événements). 213 D/ JZ/YR part 1 (extrait de « Company 91, Vol. 3 ») Le duo spectaculaire entre John Zorn (saxophone alto) et Yves Robert (trombone) a été un des moments particulièrement appréciés par le public du cru 1991 du festival255, comme en témoignent les applaudissements très nourris, et le fait que les musiciens aient immédiatement enchaîné avec une autre improvisation (également présente sur le disque). Il faut dire que nous avons ici affaire à deux véritables « maîtres » de l'improvisation libre. Nous examinerons trois larges extraits de cette improvisation jubilatoire dans l'analyse linéaire présentée ci-dessous256. 255 Grand cru, d'ailleurs. Derek Bailey en a été tellement content qu'il a publié trois disques issus de cette semaine d'improvisation. 256 Toutes les improvisations tirées de Company Weeks analysées ici se trouvent dans le CD-Rom au format MP3, dans le dossier « /Analyses/Company Weeks/ ». 1. Analyse linéaire 1) La première intervention revient au saxophoniste, et est assez typique du style zornien : courte, très aiguë et extrêmement précise (coupure très franche). Cette phrase suit une course descendante : du sur-aigu vers une hauteur distincte medium (fa). Le tromboniste répond immédiatement par une phrase qui suit une évolution inverse, donc ascendante (génération par contraste). Mais ce qui n'est encore qu'un jeu d'échange imitatif va vite se transformer en une claire répartition des rôles de type question-réponse. Le tromboniste décide en effet de créer un fort effet cadentiel à la fin de sa phrase en jouant un sol grave (écho approximatif du fa précédent) clairement séparé de l'intervention précédente par un court silence, un saut de registre d'une dixième, et une attaque sforzato. Dans ces deux premières interventions se manifeste donc déjà un certain schéma d'interaction (où le tromboniste joue un rôle conclusif) qui va rester largement prégnant pour tout le début. De même, cet effet de désinence va intervenir dans beaucoup de phrases du tromboniste au début. 2) Ici, on voit la persistance du schéma d'interaction qui s'est mis en place au tout début. Alors que celui-ci aurait pu s'inverser (c'est le tromboniste qui prend l'initiative pour la première fois, avec une sixte ascendante), c'est malgré tout le tromboniste qui produit l'effet de conclusion. 3) On observe un effet de relais : c'est sur le sol grave du trombone que le saxophoniste redémarre lui aussi sur un sol (avec effet de glissando depuis le demi-ton inférieur). Ce relais, véritable marqueur formel, va jouer comme un signal très fort pour redéfinir les rapports d'interaction entre les deux instrumentistes : au régime de la succession complémentaire (effet question-r ) va succéder celui de la simultanéité imitative (effet de synergie). Ce passage de relais porte en lui ces deux caractéristiques, par la quasisimultanéité de l'arrivée du tromboniste et du départ du saxophoniste, et par le choix de la même classe de hauteur. Dans toute la séquence qui suit, les interventions des deux instrumentistes sont tellement entrelacées que c'est plutôt le flux continu des deux discours qui devient pertinent. 4) L'arrêt quasi-simultané, au même titre que le redémarrage qui suit, sont encore des indices du type d'effet synergique que recherchent les deux musiciens. 7) Le saxophoniste procède ici à une interpolation pour se rapprocher de la proposition du tromboniste. Cela se fait tout simplement par passage progressif entre un multiphonique où domine la hauteur si à un si pur (auquel on peut ajouter l'ajustement dynamique du forte vers le piano). Notons que cela signifie que pour ce passage, le seuil d'autonomie des discours du saxophoniste est relativement bas (puisqu'il y a cet effet d'ajustement) ; et quand le seuil d'autonomie des discours d'un des improvisateurs est bas, on observe assez souvent un effet « boule de neige » qui conduit les musiciens à entretenir des rapports de similitude entre leurs discours. 8) On vérifie ce point immédiatement avec une interruption subite du tromboniste dont la proposition était dans un rapport d'autonomie avec celle du saxophoniste. Yves Robert choisit de ne pas faire d'interpolation, ce qui est assez rare, car cela fait immanquablement apparaître ce qui précède l'interruption comme une sorte d'erreur, ou de faux départ, en tout cas quelque chose de renié par le musicien. Impression sans doute également ressentie par le tromboniste qui semble ensuite hésiter entre plusieurs éléments musicaux pendant quelques secondes. Ce passage illustre également le fait qu'une des grandes difficultés de l'exigence de coordination dans l'improvisation libre est de ressentir collectivement les articulations formelles. Celles-ci se traduisent le plus souvent par des générations par interruption ; or ici, on remarque que seul le saxophoniste procède à une génération par interruption, le tromboniste effectuant quant à lui une génération par association, signe évident de sa volonté de poursuivre encore pour un moment l'idée musicale précédemment exposée. Il faut dire que cette idée musicale devait être d'autant plus saillant qu'elle venait d'être exprimée deux fois, avec une simple inversion des rôles (note tenue au saxophone et court fragment mélodique au trombone d'abord et viceversa ensuite). Un marqueur formel va permettre de mettre les deux musiciens d'accord sur le moment de l'articulation : il s'agit du si de saxophone à 109'' qui reprend, à l'octave supérieure, le si du trombone à 107''. Cet événement est d'autant plus prégnant que le saxophoniste a interrompu brutalement l'élément précédent pour introduire ce si. Le rôle de marqueur formel de ce si est définitivement confirmé quand le tromboniste le reprend encore une fois à 110''. 12) Le rapport entre les deux discours instrumentaux est, depuis le début, un rapport associatif ; tantôt par contraste (ainsi dans la configuration note-pédale/fragment mélodique ou dans la configuration question-réponse du début), tantôt par similitude (dans les passages d'imitation et/ou de fusion des deux discours). Nous sommes ici plutôt dans le deuxième cas de figure. Depuis 199'', le mode de jeu se fait similaire (débit rapide et continu, registre aigu de l'instrument, énergie élevée par la densité et la dynamique) ; le seuil d'autonomie des discours étant au plus bas (puisque les improvisateurs semblent privilégier des rapports de similitude), un changement chez l'un se traduit presque immédiatement par une modification du discours de l'autre. Ainsi, quand une quasi-pulsation apparaît sur le lab répété du trombone, le saxophoniste assèche son discours et se synchronise sur cette pulsation, puis joue de décalages avec celle-ci, désormais sous-entendue. 13) On retrouve une variante de la situation note-pédale/mélodie que nous connaissons bien. La variante vient d'un effet d'interpolation (entre le discours en slap extrêmement saccadé qui précède et l'idéal de la note tenue) qui conduit à substituer à la note tenue un slap à peu près régulier et stabilisé sur une zone de hauteurs relativement restreinte. 14) Un pur signal d'interruption au saxophone (un do unique, hauteur distincte contrastante, aussitôt interrompu) conduit le tromboniste à stopper son développement mélodique. La nouvelle séquence (polarisée autour de la septième sib-lab) naît juste après, les deux improvisateurs étant toujours dans un rapport d'association par contraste : on retrouve notamment le trombone dans son rôle de ponctuation souvent conclusive. Le développement de cette séquence obéit à un sch assez classique : chacun propose de petites variations (le chromatisme descendant au trombone, par exemple) qui amènent l'autre à répondre également par une légère variation. C'est assez net pour le saxophoniste qui modifie ainsi, de proche en proche, la nature de son « accompagnement » en bariolage, jusqu'à un point (à 275'') où la morphologie même de ses phrases se trouve modifiée ; le tromboniste saisit immédiatement ce point comme marqueur formel pour se rapprocher à nouveau du timbre du saxophoniste. 15) Encore un pur signal d'interruption, toujours au saxophone, toujours sur une note unique, cette fois-ci sur-aiguë ; il n'y a aucun doute à avoir sur la signification d'un tel geste, d'autant plus que la séquence précédente est remarquablement longue (et sur des éléments très restreints), surtout quand elle est mise en regard avec le reste de l'improvisation, d'une nature plutôt kaléidoscopique. Il joue donc idéalement le rôle de marqueur formel, et permet la remise à zéro des discours (génération par interruption pour chacun). Mais la nature de la séquence à venir, bien que tout de suite très caractérisée, ne se stabilise pas immédiatement ; nous avons ici un bel exemple de coordination progressive, où la nature du point focal (une évocation du jazz) n'émerge que progressivement, par affirmation successive. Les guirlandes du saxophone semblent au début (à 297'') assez indépendantes de la rapide walking bass du tromboniste (en témoigne la légère incertitude de la pulsation, que le musicien accélère un peu, dans l'espoir de se synchroniser avec le débit du saxophone) ; celuici n'étant pas certain de la bonne réception de son idée, il s'apprête à en changer (à 307'') et commence des traits rapides dans l'aigu de son instrument. Mais à 309'', John Zorn commence une descente à l'articulation tout à fait be-bop (swing, ornementation). A 310'', Yves Robert reprend par conséquent l'idée de la ligne de basse (plutôt binaire cette fois-ci) pendant que le saxophoniste lance une dernière guirlande virtuose. Enfin, à 312'', le saxophoniste se coordonne finalement sur la pulsation et l'univers idiomatique suggérés par le cours du tromboniste. 16) Cet enchaînement est intéressant ; en effet, la traditionnelle interruption zornienne n'a pour ainsi dire aucun impact sur le tromboniste (qui continue son discours pulsé). Mais, encore une fois, elle n'en indique pas moins clairement le désir d'un changement de section. Ce qui est remarquable ici, c'est que le changement de section se fait justement après cette interruption, de manière assez classique et naturelle, par interpolation : le débit s'accélérant, jusqu'à perdre tout sentiment de pulsation. C'est particulièrement net au trombone, qui utilise un procédé d'interpolation entre les deux régimes (pulsé et non-pulsé) : cela aboutit au faisceau entre 332'' et 335'', qui conserve la pulsation précédente mais affectée d'un léger rubato qui lui fait perdre tout son drive (et donc les connotations stylistiques afférentes). Ceci aurait très bien ) 21) pu se faire sans l'intervention de Zorn. Celle-ci a donc surtout servi à lever toute ambiguïté sur la durée possible de cette évocation du jazz, et à mettre les deux improvisateurs d'accord pour qu'ils initient une phase de transition. Toute cette section se caractérise par une très grande virtuosité : virtuosité des moyens individuels (recours à des techniques spécifiques, comme le slap, rapidité du débit) bien sûr, mais également virtuosité collective. Cette dernière se manifeste essentiellement dans les jeux de pulsation perpétuellement changeants auxquels se livrent les deux musiciens. Après le passage d'intense synergie (à partir de 312''), où le jeu pulsé a vraiment pu se développer, c'est maintenant le travail sur la pulsation elle même qui est au coeur de cette section. Ce travail prend diverses formes : pulsations contrariées (indépendantes ou quasiment pour chacun des musiciens), suggestion d'une pulsation mais immédiatement abandonnée, alternance rapide de phrases pulsées et non-pulsées, utilisation du jeu quasiment pulsé mais affecté d'un rubato et de décalages permanents. Ce jeu prend tout son intérêt dans les échanges auxquels se livre le duo, échanges qu'ils réalisent avec brio et virtuosité. On a ici un premier exemple de pulsation contrariée : le trombone installe un tempo auquel le saxophone surimpose immédiatement une valse ironique et méchante, dans son appareil le plus fruste, à la Ornette Coleman. La tension entre les deux pulsations est très grande (surtout en raison de leur proximité) mais elle finit par se relâcher quand le tromboniste se laisse gagner (à 352'') par le tempo du motif inexorablement répété par le saxophoniste. Une séquence pyrotechnique, où le jeu du trombone, encore assez discourant (et pulsé) se marie à merveille avec le flux très rythmique (non-pulsé) du saxophone, caractère provoqué par les changements incessants des modes de jeu convoqués. Voici encore un point qui illustre très bien cette joute virtuose avec la pulsation. Depuis 364'', plus personne n'a de pulsation : le discours est très sculpté chez John Zorn (phrases courtes entrecoupées de silences) et très continu chez Yves Robert (débit très dense), mais ils partagent la même énergie essentiellement ascendante. C'est justement le premier geste descendant, très en dehors (sforzato, débit beaucoup plus lent), au trombone, qui signale le changement de section. 223 encore vraiment Hauptstimme : beaucoup de notes répétées, sans véritable direction mélodique. Une note de trombone avec un peu de growl (à 421'', timbre contrastant jouant typiquement le rôle de marqueur formel) conduit le saxophoniste à assumer un rôle d'ostinato ; quelques secondes plus tard, le chant de trombone se déploie, entrecoupé de silences, d'abord polarisé sur un do duquel émergent divers sons chantés. 22) On remarque un effet d'ajustement assez typique : le changement appelle le changement. Ici (et c'est probablement dans ce sens qu'il faut le lire, même si la chronologie pourrait suggérer l'inverse) le balayage du saxophone parcourt subitement un registre plus large, faisant clairement entendre quelques hauteurs médiums ; cela pousse aussitôt le tromboniste à continuer sa pédale grave de do#, alternant une note sur deux avec le chromatisme sol#-la-sib. L'impression cadentielle est très forte, d'autant plus que le mouvement chromatique suggère une résolution imminente. Un marqueur formel (le fait que le saxophoniste termine brusquement son flux continu par une approximation de do#, un do un peu haut, donne à la coupure une saillance supplémentaire) avertit les improvisateurs que le geste conclusif peut intervenir sans ambiguïté à ce moment-là. 23) Et la cadence intervient effectivement ; non pas en do# (cela aurait été bien étonnant!) mais avec une reprise quasiment textuelle de l'ouverture de l'improvisation : un motif à la courbe ascendante puis descendante au saxophone, ponctué, avec une éloquence toute conclusive, par un fa grave de trombone. Cette fin est particulièrement réussie et virtuose car : elle est soudaine et surprenante (elle vient interrompre une des plus longues sections de l'improvisation ) ; elle est parfaitement réalisée (pas d'hésitation, de notes qui débordent, d'incompréhension de la part d'un des musiciens) ; elle dote l'improvisation d'une clôture formelle ; elle est absolument sans ambiguïté, en témoignent les applaudissements quasi-immédiat du public qui en saisit donc toute la signification. On sait la difficulté de terminer une improvisation libre, en l'absence de tout référent externe ; ici, on voit que le motif en question s'est constitué comme point focal, d'abord parce qu'il a eu le privilège d'ouvrir l'improvisation, ensuite parce qu'il a été répété plusieurs fois (on a vu que la première section était bâtie sur cette répartition des rôles de type question-réponse). 2. Synthèse : Hyper-séquençage, rôle des interruptions et saillance des schémas d'interaction Cette improvisation s'étend sur un peu plus de 8 minutes. Or, la première chose que l'on remarque, c'est qu'on peut facilement la découper en un assez grand nombre de séquences, 11 à notre sens, ce qui fait une moyenne de 45'' par séquence : 1) Du début à 61'' : une séquence allant progressivement d'échanges aérés à une pure synergie, le tout dans un style très virtuose (débit rapide, alternance extrêmement rapide de modes de jeux et de registres). 2) De 61'' à 111'' : une séquence dominée par des fragments mélodiques au trombone, le saxophoniste prenant un rôle d'accompagnateur excentrique. 3) De 111'' à 153'' : une séquence très humoristique (rires du public) avec caquètements, cris et bruits en tout genre. 4) De 153'' à 199'' : une séquence « folklorique » : musique des îles au trombone et évocation du Sacre au saxophone. 5) De 199'' à 237'' : une séquence proche de la première, alternant effets de synergie (le passage en slap) et échanges hyper-virtuoses. 6) De 237'' à 250'' : une séquence très courte, avec nouvelle évocation folklorique au trombone. 7) De 250'' à 295'' : une séquence construite autour de bisbigliando au saxophone, également humoristique. 8) De 295'' à 344'' : une séquence très « jazz », avec walking bass et solos. 9) De 344'' à 389'' : une séquence de quasi-valse, faisant furieusement penser à du Ornette Coleman, se dissolvant dans le chaos. 10) De 389'' à 426'' : une séquence dans les dynamiques piano, avec beaucoup d'effets instrumentaux (slaps, bisous). 11) De 426'' à la fin : une séquence où le tromboniste chante dans son instrument, avec en guise d'accompagnement un babil aigu de saxophone. On pourrait sans doute proposer un découpage encore plus fin, pour rendre compte de la nature kaléidoscopique de cette musique, mais ce serait en fait donner l'illusion d'une improvisation formellement très instable, ce qui n'est nullement le cas. Il nous semble au contraire que la lisibilité de cette improvisation est particulièrement grande. On peut donner au moins deux raisons à cette très grande richesse séquentielle : d'abord, celle-ci fait partie intégrante de l'impression générale de virtuosité recherchée par les musiciens (elle n'en est qu'une manifestation à une plus grande échelle) ; ensuite, la forme du duo permet (et peut-être même demande) une grande souplesse formelle pour une raison évidente : la coordination y est beaucoup plus aisée. Pour le dire de manière triviale, il est plus rapide de mettre deux personnes d'accord qu'un large groupe. On peut sans doute approfondir ce dernier point. En fait, il ne nous semblerait pas faux de dire que le duo exige (et pas seulement permet) effectivement une plus grande variété séquentielle. Cela est lié au critère de lassitude qui commande dans une large mesure le besoin de changer de séquence. Qu'est-ce qui fait augmenter la lassitude? D'abord la perpétuation de certaines caractéristiques acoustiques, cinétiques ou musicales. Mais aussi la permanence d'un certain schéma d'interaction. Concernant ce deuxième point, la lassitude n'augmente pas toujours 225 uniformément : ainsi, on peut supposer qu'un schéma d'interaction très différencié260 provoque moins de lassitude pour les improvisateurs qu'un schéma d'interaction très uniforme. Or, dans le cas du duo, les rapports étant le plus souvent symétriques, on voit bien que le système d'interaction ne peut pas être très riche : d'où une plus grande lassitude pour les improvisateurs. Ce point est donc très important pour comprendre l'hyper-découpage de l'improvisation qui nous occupe. Pour autant, on l'a dit, on a le sentiment d'une grande solidité formelle. C'est qu'ici, et la réussite vient en grande partie de là, le changement de séquence n'est pas subordonné au changement du schéma d'interaction ; autrement dit, un certain nombre de caractéristiques acoustiques, cinétiques ou musicales peuvent persister malgré la modification du rapport d'interaction (qui imitait plutôt l'autre, qui était très autonome), conférant par là son unité à la séquence. Ce n'était pas le cas dans l'improvisation des Emeudroïdes que nous avons examinée au début de cette partie : on pourrait donc faire l'hypothèse que plus le nombre d'improvisateurs est élevé plus il y a des chances pour que le schéma d'interaction soit prédominant dans la définition de la séquence. La deuxième chose importante qu'il faut signaler concerne la manière dont se font les articulations entre les différentes séquences. Dans la très grande majorité des cas, c'est un signal d'interruption (très souvent du saxophoniste) qui provoque le changement de séquence (voir les points 14 et 15 par exemple). La logique du marqueur formel se trouve donc poussée jusqu'à l'extrême : ces interruptions n'ont pas d'autre fonction que métapragmatique, c'est-à-dire qu'elles servent à modifier le cadre général dans lequel doivent prendre place les interactions ; mais elles sont absolument vides de contenu. Ce sont de purs signaux qui, par leur saillance évidente (un son très fort et très aigu), permettent de mettre les improvisateurs d'accord sur les moments de bascule, d'articulation, de changement. La réactivité des musiciens n'en est pas moins vraiment remarquable, avec des re-coordinations parfois presque instantanées (comme à 295''). C'est évidemment de ce procédé que découle la très grande lisibilité formelle de l'improvisation, avec de très nombreux enchaînements « cut », à mille lieux des transitions progressives et des processus d'interpolation. chose d'autre doit commencer. On peut être agacé par cette attitude, parfois peu « démocratique », mais il faut bien reconnaître que c'est une des spécificités les plus intéressantes du style d'improvisation de John Zorn. De son côté, Yves Robert n'hésite pas à plusieurs reprises (à 237'' par exemple) à amorcer une nouvelle séquence de manière unilatérale, dans un geste forcément très interruptif. Néanmoins, la logique reste différente, puisque ces gestes, pour interruptifs qu'ils soient, sont aussi plus que des signaux : ils portent en eux un contenu (acoustique, cinétique ou musical) qui est définitionnel de la séquence qui s'amorce. La première intervention « swinguée » de Zorn à 61'' (voir point 5) est également de cette nature-là. Si l'on résume, les articulations sont donc essentiellement de nature interruptive : 1) Ou bien parce qu'il y a effectivement un signal d'interruption qui met tout le monde d'accord : il faut commencer quelque chose de nouveau. 2) Ou bien parce qu'un des musiciens décide unilatéralement d'introduire un nouvel élément très contrastant, qui a donc une grande probabilité de se retrouver au coeur de la séquence à venir. Bien sûr, certaines articulations utilisent également des marqueurs formels plus traditionnels, comme au point 8 (transition de la séquence 2 à la séquence 3), avec l'apparition d'un si commun, confirmé à nouveau par le tromboniste, levant ainsi toute ambiguïté et permettant aux musiciens d'attaquer une nouvelle séquence, très humoristique, de grande synergie. Enfin, il convient de signaler un dernier point, qui concerne les rapports d'interaction à l'oeuvre dans cette improvisation. On peut en distinguer trois principaux : des rapports de synergie (imitation récipro visant à donner l'impression d'un discours et/ou d'un instrument unique) ; des rapports soliste/accompagnateur (développement mélodique/tenue ou texture ou phrase répétée) ; et des rapports d'association contrastée (type question-réponse). Il est intéressant de noter que ces rapports apparaissent tous les trois au début de l'improvisation : au bout d'un peu plus d'une minute, on a successivement le dialogue-joute (points 1 et 2), la synergie (points 3 et 4) et le solo/accompagnement (points 5 et 6). Une fois ces trois possibilités établies, les musiciens parviennent à les convoquer à nouveau dans le cours de l'improvisation avec beaucoup de facilité. Le cas le plus clair est sans doute celui du rapport soliste/accompagnateur. First (extrait de « Epiphanies ») L'étude de cette improvisation (du moins de ses sept premières minutes) du festival de 1982, un duo entre la pianiste Ursula Oppens et le contrebassiste Moto Yoshizawa, a bien pour but de mettre en lumière certains mécanismes de coordination, mais seulement de manière indirecte. En effet, à notre sens, il s'agit d'une improvisation franchement ratée, et en même temps très caractéristique des travers dans lesquels peut aisément tomber l'improvisation libre. Ben Watson, dans la longue section qu'il consacre aux divers Company Weeks, expédie assez rapidement cette partie, qui constitue la première plage du disque Epiphanies : « Dans leur duo, Oppens et Yoshizawa échouent passablement à se connecter, produisant une sorte de musique brisée dont « l'imprévisibilité » faisait les délices de Derek Bailey, mais qui peut rebuter l'acheteur de disques261 » (Watson 2004, p. 234). Cette absence de connexion soulignée par Watson, nous la comprenons comme échec de coordination. Nous allons donc essayer de montrer comment, et pourquoi, cette improvisation produit sur l'auditeur cette impression défavorable, impression qui n'est peut-être pas tant due à l'imprévisibilité du contenu mais bien à sa trop grande prévisibilité. 261 « In their duet, Oppens and Yoshizawa utterly fail to connect, providing just the kind of broken music Bailey delights in as « unpredictable », but which can disappoint the record buyer ». 1. Analyse linéaire 1) La première entrée des deux instrumentistes est déjà tout à fait caractéristique du schéma interactif qui irrigue toute cette improvisation : impulsion-réaction. Une description en termes behavioristes (stimulus-réflexe) ne serait d'ailleurs peut-être pas tout à fait hors de propos, étant donné que bien souvent la réaction semble davantage intervenir au plan gestuel de l'émission du son, qu'au plan musical proprement dit (nous en donnerons quelques exemples peu après). Cette impression de réflexe est encore renforcée par la très grande brièveté des propositions, proches d'impulsions acoustico-gestuelles. Si le premier échange se caractérise par un effet de complémentarité (opposition forte/piano, motif brisé/glissando continu), les échanges suivants vont devenir plus imitatifs : similitudes morphologiques, tentatives pour trouver des équivalents de timbre (ainsi entre 4'' et 7'', avec les effets de percussion sur la table d'harmonie du piano et les col legno de la contrebasse) Il s'agit bien d'échanges sur le modèle du dialogue, c'est-à-dire que la simultanéité est évitée, même si parfois quelques résonances ou impulsions conclusives de l'un sont encore présentes sur l'entrée de l'autre. 2) Ce pizzicato grave au piano (fa) ne déclenche rien et apparaît rétrospectivement comme un « coup pour rien ». Bien qu'il soit nettement saillant, d'abord pour la raison simple qu'il est isolé de part et d'autres par des silences (qui le mettent en valeur), ensuite parce qu'il s'agit d'une hauteur distincte émise de manière relativement pure, il ne sert pas de prétexte à développer ou bien varier le discours. Témoin la reprise à 22'' du schéma impulsion-réaction. Deux hypothèses possibles : ou bien ce pizzicato était un geste non volontaire (mais cela semble peu probable vu le silence qui le précède), de type accident, et qui reste non-assumé, accepté comme erreur, et donc renié par le glissando suivant qui permet de revenir au dialogue précédent ; ou bien il s'agit bien d'un geste volontaire introduisant un nouvel élément dans l'improvisation, mais devant l'absence de réaction du partenaire (le silence qui suit), il reste en friche puisque immédiatement renié. 3) Voici un exemple net d'ajustement quasi-moteur du discours du piano sur celui de la contrebasse. Après un glissando de relance dans l'aigu et piano, Ursula Oppens s'aligne immédiatement sur l'entrée du contrebassiste (avec un son écrasé) en produisant immédiatement un nouveau glissando beaucoup plus serré (presque arraché) et plus fort. 5) On peut noter un autre trait assez systématique de cette improvisation : le passage par le silence absolu entre les différentes séquences de l'improvisation. Ces silences ont une double caractéristique : d'abord, il ne s'agit pas de simples respirations, mais plutôt d'arrêts dans la production musicale (ce que traduit la durée de ces silences, parfois fort longs, comme c'est le cas ici) ; d'autre part, c'est presque toujours de la même courbe dynamique qu'il s'agit : diminuendo-silence-reprise piano (même s'il y a bien sûr des exceptions). Ces deux caractéristiques font que les silences sont vécus par l'auditeur (et sans doute aussi par les musiciens) comme un temps d'attente qui brise et fragmente sans cesse l'élan d'un discours qui ne démarre véritablement jamais. Cela est d'autant plus flagrant que les diminuendi amenant au silence ne sont pas vraiment des diminuendi conduits musicalement mais des affaissements dynamiques résultant d'interventions souvent hésitantes, et donc non-investies, à l'énergie très faible (on a noté les pizzicati de piano isolés ; ici, à 37'', c'est d'un gratouillis dans le piano, peut-être effectué à l'aide d'une baguette de percussion qu'il s'agit). 6) Évidemment, cette impression vient également du fait que les propositions avortent très rapidement : c'est le cas ici. En effet, il semble qu'un élément nouveau apparaisse à partir du trille fa#-fa de piano (même si on peut l'interpréter comme continuation du battuto de la contrebasse à 37''). Il s'agit clairement d'un marqueur d'articulation formelle pour les deux improvisateurs : d'abord par l'effet de timbre produit c'est la première fois que l'on entend le son du piano produit par l'action des marteaux), ensuite par le principe de répétition contenu dans l'idée même de trille (ou de mordant, pour être exact). Il serait donc tout à fait propice à l'enclenchement d'une nouvelle séquence. Et c'est bien ce qui se passe. Mais d'abord, cela se fait de manière assez timide, et non franche. La raison en est due au systématique effet d'ajustement qui conduit la pianiste à n'entretenir que des rapports de similitude avec le discours du contrebassiste. Ainsi, après ce trille, le contrebassiste répond (42'') par un battuto col legno (qui reprend l'idée de répétition du trille) mais dans une dynamique très piano ; aussitôt (44''), la pianiste reprend par deux notes égrenées pianissimo, fa#-sol, qui donneront finalement naissance à un nouveau trille (45''). 9) La pianiste interrompt clairement la séquence initiée par le contrebassiste (comme quelqu'un qui est tombé du train en marche). Encore une fois, l'auditeur a donc un sentiment d'une séquence brisée, avortée, abandonnée avant même d'avoir eu une chance de s'épanouir. 10) Le contrebassiste n'abandonne pas la partie et, après une courte réponse à l'interruption précédente, poursuit la même idée (présentée différemment), sans doute dans l'espoir que sa partenaire se coordonne correctement avec cette stratégie. Pourtant, la pianiste répond encore une fois avec le même type de proposition sonore (un glissando sur les cordes du piano, mais purement bruitiste cette fois-ci). Le contrebassiste évolue alors vers la pure trame : d'abord, en ne faisant plus entendre que le bruit de l'archet frotté ; puis en partant sur un trémolo, très piano, avec également beaucoup de bruit d'archet. Cette production de texture est assez nettement une stratégie d'accompagnement, appelant l'autre à développer un discours plus clairement en dehors. Mais la pianiste se contente de deux interventions très brèves (la deuxième peut-être involontaire, d'ailleurs) dont le but est clairement de se fondre dans la texture globale. La première intervention à 90'' s'inscrit dans le crescendo initié par le contrebassiste une seconde avant ; elle provoque d'ailleurs chez le contrebassiste un ajustement dynamique immédiat avant que celui-ci ne revienne subitement à une dynamique pianissimo. 11) Le passage de relais dans la texture est ici assez réussi. L'entrée du piano sur des pizzicati répétés (rapport de similitude avec le discours du contrebassiste) conduit ce dernier à prendre une parole « soliste » (même si 'est de manière très discrète et discontinue) ; mais il ne fait pas cela immédiatement : n'oublions pas que cette improvisation manque cruellement de continuité et donc que toutes les occasions où l'on peut réaliser ce genre de tuilage sont sans doute bonnes à prendre. Il achève d'abord sa production texturale par un diminuendo, puis énonce clairement un geste rythmique (percussion sur la caisse de résonance) qui s'inscrit dans une pulsation suggérée très brièvement par la pianiste au début de son entrée. Ce geste pourrait sans doute jouer le rôle d'un marqueur formel clair (saillance de la pulsation) mais la section échoue encore une fois à se développer, la pianiste arrêtant de gratter les cordes à 114'' et laissant le contrebassiste seul.	45,275
https://openalex.org/W2804122997	OpenAlex	Open Science	CC-By	2,018	A citizen science based survey method for estimating the density of urban carnivores	Dawn M. Scott	English	Spoken	8,783	15,488	RESEARCH ARTICLE A citizen science based survey method for estimating the density of urban carnivores Dawn M. Scott1, Rowenna Baker1, Naomi Charman1, Heidi Karlsson1, Richard W. Yarnell2, Aileen C. Mill3, Graham C. Smith4, Bryony A. Tolhurst1 1 Conservation Ecology and Zoonosis Research Group, University of Brighton, Brighton, United Kingdom, 2 School of Animal, Rural and Environmental Sciences, Nottingham Trent University, Brackenhurst Campus, Southwell, United Kingdom, 3 Centre for Wildlife Management, Newcastle University, Newcastle upon Tyne, United Kingdom, 4 Animal and Plant Health Agency, Sand Hutton, York, United Kingdom dawn.scott@brighton.ac.uk a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS Citation: Scott DM, Baker R, Charman N, Karlsson H, Yarnell RW, Mill AC, et al. (2018) A citizen science based survey method for estimating the density of urban carnivores. PLoS ONE 13(5): e0197445. https://doi.org/10.1371/journal. pone.0197445 Editor: Govindhaswamy Umapathy, Centre for Cellular and Molecular Biology, INDIA Editor: Govindhaswamy Umapathy, Centre for Cellular and Molecular Biology, INDIA Editor: Govindhaswamy Umapathy, Centre for Cellular and Molecular Biology, INDIA Received: August 24, 2017 Accepted: May 2, 2018 Published: May 22, 2018 Copyright: © 2018 Scott 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. Received: August 24, 2017 Accepted: May 2, 2018 Published: May 22, 2018 p p p Using species-specific clustering units that reflect social organisation, the method was addi- tionally applied to suburban badgers to derive relative badger group density (BGD) for one city (Brighton, 2.41 km-2). We demonstrate that citizen science approaches can effectively obtain data to assess suburban carnivore density, however publicly derived national data sets need to be locally validated before extrapolations can be undertaken. The method we present for assessing densities of foxes and badgers in British towns and cities is also adaptable to other urban carnivores elsewhere. However this transferability is contingent on species traits meeting particular criteria, and on resident responsiveness. Copyright: © 2018 Scott 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 cannot be made publicly available for ethical reasons - in this case addresses/spatial locations of participants are to be kept private and this was stated in the letter to participants during the study. Secondly, data deposition could present some threat to protected species, in this case European badgers, that are a protected species. The authors feel that releasing location data of badger setts could lead to a potential threat. Data can be available via chair of the School Ethics committee (F.Guppy@brighton. Abstract Globally there are many examples of synanthropic carnivores exploiting growth in urbanisa- tion. As carnivores can come into conflict with humans and are potential vectors of zoonotic disease, assessing densities in suburban areas and identifying factors that influence them are necessary to aid management and mitigation. However, fragmented, privately owned land restricts the use of conventional carnivore surveying techniques in these areas, requir- ing development of novel methods. We present a method that combines questionnaire dis- tribution to residents with field surveys and GIS, to determine relative density of two urban carnivores in England, Great Britain. We determined the density of: red fox (Vulpes vulpes) social groups in 14, approximately 1km2 suburban areas in 8 different towns and cities; and Eurasian badger (Meles meles) social groups in three suburban areas of one city. Average relative fox group density (FGD) was 3.72 km-2, which was double the estimates for cities with resident foxes in the 1980’s. Density was comparable to an alternative estimate derived from trapping and GPS-tracking, indicating the validity of the method. However, FGD did not correlate with a national dataset based on fox sightings, indicating unreliability of the national data to determine actual densities or to extrapolate a national population estimate. Using species-specific clustering units that reflect social organisation, the method was addi- tionally applied to suburban badgers to derive relative badger group density (BGD) for one city (Brighton, 2.41 km-2). We demonstrate that citizen science approaches can effectively obtain data to assess suburban carnivore density, however publicly derived national data sets need to be locally validated before extrapolations can be undertaken. The method we present for assessing densities of foxes and badgers in British towns and cities is also adaptable to other urban carnivores elsewhere. However this transferability is contingent on species traits meeting particular criteria, and on resident responsiveness. * dawn.scott@brighton.ac.uk Assessing urban carnivore density successfully exploited the ecological opportunities associated with urban development, becom- ing synanthropic such that their densities in urban habitat typically exceed those in natural landscapes [2, 3, 4]. In Europe examples of synanthropic species include the red fox (Vulpes vulpes) and Eurasian badger (Meles meles) [2]. Carnivore adaptations to urban environments include ecological and behavioural changes [5], which combined with increasing density can affect the type and magnitude of intra- and inter- specific interactions. Although urban wildlife can provide benefits to human residents [6, 7] the occurrence of high density carnivore popu- lations in close proximity to humans raises concerns about potential conflicts [8, 9] including increased risk of zoonotic disease transmission [8, 10]. Therefore assessing urban carnivore density, understanding those factors which influence density, and determining patterns of temporal change are essential for devising strategies for management, mitigating conflicts and controlling zoonotic disease [11, 3]. ac.uk) if ethical and confidential restrictions of the data set are met. Funding: The project was partially funded by Earthwatch Institute (earthwatch.org), Urban Mammals Project Grant (DS) for the first year. This funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The University of Brighton (DS) and APHA (DS) provided supplementary funds to cover equipment and travel costs. Universities of Nottingham Trent and Newcastle supported undergraduate students involved with the project data collection at respective cities. Academics at these institutes had a role in study design, data collection and analysis, decision to publish, and/or preparation of the manuscript. Globally, the red fox is the most widespread and successful urban carnivore, having colo- nized cities in Europe, Australia, USA, Canada and Japan [12, 13, 14]. In Europe, it is host to a range of diseases and parasites including the zoonotic tapeworm Echinococcus multilocularis and rabies (Lyssavirus sp.) [15, 16]. Although foxes in Britain are currently free of E. multilocu- laris, this causative agent of human alveolar echinococcosis (HAE) is becoming more widely distributed in Europe, and continued surveillance is recommended [17]. Likewise, it is possible that rabies could be re- introduced to Great Britain and the occurrence of both red foxes and Eurasian badgers at high density has the potential to sustain a major epizootic [18]. Effective control of such an outbreak would require accurate estimation of urban fox and badger social group densities (hereafter termed FGD and BGD respectively) [18]. Introduction Urbanization is a major cause of land use change worldwide, typically resulting in the loss of biodiversity [1]. Conversely, several behaviourally flexible mammalian carnivores have 1 / 14 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 Study sites Urban fox sightings were previously collected from members of the public during April and May 2012 and used to calculate fox sightings density (FSD: the number of sightings per 1000 people km-2) for 144 towns/cities across Great Britain (for full methodology see [24]). How- ever Scott et al. (2014) state that follow up on the ground field studies in a sub-set of cities should be conducted to quantify the relationship between ‘true’ densities relative to sightings density. Based on the FSD values, eight towns/cities in England deemed representative of FGD range in British cities were selected for the current study. Because of an historic north-south divide in urban fox distribution [36] four towns/cities were selected from northern and eastern England (Newcastle, Huddersfield, Preston, and Norwich) and four from the south (London, Bournemouth, Portsmouth and Brighton). To investigate variable FGD within suburban areas, more than one site was surveyed in four cities where such replication was logistically feasible, according to FSD categories (low-high) as follows: Huddersfield (N = 2 sites; low, FSD = 14.6), Brighton (N = 4; medium FSD = 37.8), Newcastle (N = 2; medium FSD = 51.2) and London (N = 2; high FSD = 703.1). All other cities were N = 1 due to survey logistics. In parallel with fox data collection, badgers were also surveyed in one city (Brighton, N = 3) that had known occurrence of badgers and a historical density estimate [5]. Blocks of approximately 1km2 were randomly selected at each site within each town/city. This size was deemed large enough for potential occupation by multiple social groups, but logistically feasible to survey within a narrow temporal window. Assessing urban carnivore density Badgers typically use one main sett, plus annex setts and outlying holes [31]. A positive association between number of main setts and number of social groups forms the basis of bad- ger social group density estimates in rural Britain [34]. Main sett counts combined with radio- tracking have been used to determine urban densities locally [5]. However wider transferability of main sett counts to estimate BGD within urban areas is challenging as the size and spatial configuration of urban badger group territories differ from rural ones [5, 35]. Therefore, an independent method of delineating urban social groups would be beneficial. Ideally, the method would be low-cost, implementable within suburban constraints and adaptable to other suburban carnivores and localities, potentially leading to rapid multi-carnivore assessments. We used a combined questionnaire, field survey and GIS-based method to survey urban foxes and badgers. We aimed to: (i) determine and contrast current relative FGD estimates in suburban areas of selected towns/cities in England; (ii) compare these with historical estimates to determine any temporal changes; (iii) investigate if suburban landscape composition pre- dicts FGD; and (iv) determine whether FGD correlates with relative sightings density of foxes across Britain from a previous dataset [24]. Furthermore, we explore the potential application of the method to another urban carnivore (Eurasian badger). Finally, we discuss the method’s wider applicability to other urban carnivores. Rabies contingency plans for Britain developed in the 1980s were reliant on the relationship between FGD and human sociological data demonstrated at the time [19, 20]. However, in response to a growing human population, urban areas have expanded and urban landscape structure changed [21], as has the socioeconomic profile of residents and their interactions with wildlife [8, 7]. Simultaneously, urban fox abundance and distribution has altered [22, 23, 24, 25] and consequently there is a need to assess current FGD. Urban badgers are less widespread globally but increasing reports in Britain [2] concurrent with incidences of conflict [9] highlight the need to also assess badger density. Competing interests: The authors have declared that no competing interests exist. In general, foxes and badgers are most frequently found in residential, suburban areas [26, 5] occurring less frequently in city centers and industrial zones [27, 28]. However the patch- work of privately owned land in suburban areas restricts the use of conventional carnivore sur- veying techniques such as direct sightings from spotlighting, or indirect quantification of field signs [29]. However, the presence of residents lends itself to the use of questionnaire surveys and “citizen science” based approaches which have been used successfully to monitor urban mammals [30, 8]. Both foxes and badgers typically live in social groups that defend exclusive territories and seasonally produce one litter per year [31, 13]. As social group size varies considerably in both species, density is typically expressed as social group density. Harris [32, 33] devised a survey technique to determine urban FGD based on sightings by schoolchildren of litters of cubs, to determine the number of social units. Additionally, mean territory size was estimated from local radio-tracking and mean group size from capture-mark recapture studies. This method was subsequently used in several urban areas to derive data for populating predictive models to determine FGD in multiple cities in Britain [27]. However, feasibility of using school chil- dren as assessors varies locally and the expertise and expense required to trap and radio-track animals limits the method’s wider applicability [29]. Cub sightings to determine fox social groups and density have also been applied outside of Britain, for example in Melbourne, Aus- tralia [12]. PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 2 / 14 Home range, buffer size and alternative estimates Fox cubs typically emerge from their natal den aged 4–6 weeks (mid-April to May). Up until 12–14 weeks old (July) the area over which they range gradually increases to include a series of secure locations (“rendezvous sites”) typically spaced <200m apart [37]. Where territories encompass multiple litters, these are often pooled such that individual cubs with different mothers will be present at the same rendezvous sites [37, 38]. One consequence of this behav- iour is that all the cubs present within one social group may be observed by multiple house- holders at multiple sites in any given year. A badger social group typically uses one main sett with annex setts and outlier holes, although when studying urban badgers in Brighton, Davi- son et al., (2008) found that annex holes were usually within 150m of the main sett. Conse- quently, for both species, it was necessary to devise a methodology for merging multiple records into a single cluster representing a social group. To inform an appropriate buffer size for merging records, the mean 95% home ranges of urban foxes were determined from parallel GPS-tracking studies in Brighton [39]. Between 2012 and 2015, the authors tracked 20 foxes (12 males; 8 females) from different social groups and derived kernel density estimate (KDE) home ranges using at least 200 relocations per ani- mal (for full field and computational methodology see [39]). Between 2014 and 2015, 6 badgers were trapped and GPS-tracked in Brighton (H. Karlsson, unpublished data) following a modi- fied fox-trapping protocol in accordance with Natural England (NE) Guidelines. Procedures were conducted under Home Office licence PPL 7007429, and NE licence 2014-3671-SCI-SCI. All protocols were approved by the ethics committee of the School of Pharmacy and Biomolec- ular Sciences, University of Brighton. A 200m buffer distance was selected for both species for merging observations based on: mean radial distances of 95% KDE home ranges (foxes = 14.2ha [±3.26 SE]; badgers = 6.53ha [±1.91 SE] (H. Karlsson, unpublished data); mean distance moved by fox cubs between ren- dezvous sites in July [37]; mean distance between main and annex setts within an urban badger social group; and main sett dispersion of <570m [35]. To validate FGD estimates, an addi- tional density calculation method from field data was employed at the Elm Grove site (Brigh- ton). Questionnaires and field surveys Surveys were conducted between July and mid-August in 2013, 2014 and/or 2015. Question- naires were distributed by hand to all residential buildings within each of the study sites, excluding those stating “no junk mail” (<0.1% of houses surveyed). Schools, hospitals, busi- nesses and other buildings also received questionnaires to cover their grounds. To increase return rates, questionnaires included either a pre-paid return envelope or an institute-specific email address. All questionnaires additionally included an identical online survey link. In Huddersfield and Newcastle, questionnaires were combined with door-to-door enquiries to PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 3 / 14 Assessing urban carnivore density increase return rates. Questionnaires requested the respondent’s address, and asked whether they had sighted any fox cubs on their residence between April and July in the current and pre- vious year. At three sites in Brighton we additionally asked residents to report badger sightings in their gardens and surrounding areas in the current and previous year and the location of any known badger setts in the area. Questionnaire data were submitted anonymously and vol- untarily. Concurrently we undertook an active search of all the public green spaces and acces- sible unpaved ground within the 14 sites to locate any active fox earths. In Brighton, badger setts were also surveyed and sett type (main or other) recorded. Home range, buffer size and alternative estimates GPS-tracking of neighbouring fox social groups was used to determine number of group territories, and territory size and configuration (see [39] for methods of live trapping and GPS tracking). During live trapping foxes were individually identified using external features and face markings. In addition camera traps (Bushnell, Kansas, USA) were placed at two locations at least 100m apart within each home range and operated for a minimum of seven nights. Group size was based on the minimum number of fox individuals recorded within the home range from both live trapping and camera trapping. Group size was based on the minimum number of fox individuals recorded within the home range from both live trapping and camera trapping. GIS analysis Questionnaires were checked for errors and any that were incomplete or obviously erroneous excluded. All questionnaires with valid addresses and reports of fox cubs, badger sightings or PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 4 / 14 Assessing urban carnivore density badger setts within the study area were spatially referenced using Ordnance Survey (OS) grid reference finder (http://gridreferencefinder.com). To accurately record the area that had been surveyed, all reported locations were plotted and a 50m buffer created around each return. Buffer size was based on local field experience: we considered 50m (equivalent to approxi- mately two gardens in an area of semi-detached housing) a suitable distance within which a householder was likely to be aware of fox cub or badger sett activity. However, this buffering approach created a concave outline with cavities that were partially surrounded by gardens from which data were available. In the case of foxes, which occupy convex territories [26], such “pockets” would be likely to constitute part of the territory hence excluding them would result in underestimating the area surveyed, and over-estimating FGD. Consequently, concavities were integrated into the survey area in cases where at least 75% of the surrounding edge was covered by survey respondents and/or surveyed greenspaces from field data validation. All fox cub records, badger sightings and setts spaced less than 200m apart were integrated into a single centroid point using the Integrate tool within ArcGIS (v 10.3.1). An additional 200m buffer area was then added around these points, and merged with the survey boundary to produce an overall survey area. Size of survey area (km2) was calculated using the ArcGIS Calculate Geometry tool and the number of independent centroids summed to estimate num- ber of social groups within the survey area (See Fig 1). Temporal changes in suburban fox occupancy were measured in terms of time since coloni- sation for each town/city: either ‘recent’ if resident foxes were present after 1986 only; or ‘long- term’ if present before 1986 [19]. England covers an area of 130,279 km2, of which 9% (11,690 km2) is classified as ‘urban’ [40] and 7% (9,116.4 km2) ‘suburban’. Suburban areas contain 55% of the human population in England and Wales [39]. To determine whether suburban landscape structure influenced FGD, housing density and the spatial extent (area) of green space, residential gardens, Fig 1. Fox cub centroid integration. PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 Statistical analysis To determine whether FSD from Scott et al.’s (2014) national fox sightings study could consti- tute a proxy for FGD and thus allow extrapolation to other cities using estimates from compa- rable urban areas, we explored the relationship between these two non-normally distributed variables using Spearman’s rank correlation. To identify landscape variables potentially pre- dicting FGD, we scrutinized five continuous variables (green space [GS], garden area [GA], housing density [HD], industrial area [IND] and made ground [MG]), and one binary variable (time since colonisation [COLON]: recent or long-term). FGD was square-root transformed to stabilise variances and consequently modelled with a Gaussian error distribution and a log link function. Collinearity between predictors was checked post- analysis using a Variation Inflation Factor (VIF) procedure, where VIF coefficients were calculated for each variable, and the largest value removed sequentially until all remaining values were within acceptable bounds (i.e. VIF <2.5). This resulted in the removal of garden area as a candidate model term (VIF = 45.67). As multiple densities were calculated for some towns/cities, we included CITY, with 8 levels, as a random factor in our Generalised Linear Mixed Model (GLMM). A back- wards stepwise model selection procedure was employed where all fixed effects were initially entered together before sequential removal of non-significant variables at the 95% level, based Table 1. Land use types, features included and excluded and spatial statistics. Land use type Inclusions Exclusions Spatial Statistic Residential Dwellings • Houses • Flats • Care homes • Halls of residence • Sheds and garages • Churches • Industrial buildings • Train stations • Shopping centres • Incomplete dwellings resulting from clipping OSMM to survey boundary Housing density No. houses km-2 (HD). Residential Gardens • Gardens of residential dwellings • Walled areas around flats • Church yards • School grounds • Incomplete gardens resulting from clipping OSMM to survey boundary Garden area km-2 (GA) Urban Green Space • Amenity grassland/parks • Extensive road verges/islands • Church yards • School grounds • Allotments • Sports grounds • Railway embankments • Hard ground fenced tennis courts Urban green space km-2 (GS) Made Ground • Manmade Surfaces: • Roads and verges • Car parks • Tennis courts • School playgrounds • Any natural environment within these Made ground km-2 (MG) Land use types with features included and excluded per category and spatial statistics derived from each survey area. OSMM–Ordnance Survey Master Map. GIS analysis An example of the integration method to derive the number of independent fox social groups from cub sightings for one survey site. https://doi.org/10.1371/journal.pone.0197445.g001 Fig 1. Fox cub centroid integration. An example of the integration method to derive the number of independent fox social groups from cub sightings for one survey site. https://doi.org/10.1371/journal.pone.0197445.g001 5 / 14 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 Assessing urban carnivore density industrial area and manmade surfaces were calculated per km2 within the survey area from Ordnance Survey master map topography (https://www.ordnancesurvey.co.uk/business-and- government/products/topography-layer.html). Suburban landscape variables used in the study are described in Table 1, listing features included and excluded and the spatial statistics derived from each survey area. The spatial extent of each land use type was validated using satellite imagery (Bing maps and Google Earth). Results Of ~30,000 questionnaires distributed across a total survey area of 17km2 across eight towns/ cities, 5645 responses (19%) were received; an average response of 403 questionnaires per study area (n = 14, SE = 61.3). This resulted in 1802 fox cub sightings. Field studies verified locations reported from the questionnaires but did not add additional cub sightings to the data. Using cub sightings integrated at 200m, mean FGD was 3.63 km-2 (95% CI of mean: 3.11–4.59) (Table 2). The alternative density estimation method used in 0.72km2 of Elm Grove (Brighton) gave a FGD of 4.17 km-2 and an individual fox density of 13.9 foxes km-2 if multi- plied by the local mean group size of 3.33 adults (n = 3). This was broadly similar but slightly higher than that derived using cub sightings integrations in a nearby, and partially overlapping location (FGD = 3.39 km-2, individual density = 11.5 km-2 if multiplied by average group size 3.4 following [41]). Precision of replicates was relatively high in Brighton and Huddersfield but lower in Newcastle and London (Table 2). Of the seven towns/cities with historical esti- mates for fox social groups [36] four that had previously not supported any established groups now did so, and the remaining three showed an increased FGD. The magnitude of this increase varied. In cities which had previous established fox populations (London, Brighton and Portsmouth) the average FGD had doubled from 1.73 to 3.51 km-2. In six areas that had not previously supported any groups, current mean FGD was 3.71 km-2. If densities in these Table 2. Fox and badger group density and landscape composition of study sites. City Region Area surveyed (km2) No. of response Green space (km-2) Garden area(km-2) Housing density (km-2) No. cities with recent colonisation No. of response = number of respondents data used in analysis, No. of fox cub records = number of fox cub records in one site over 2 years. FSD = Fox sightings density from [24]; (previous) = values in parenthesis denote predicted fox social group densities from [36]. Statistical analysis Any additional areas or exclusions were identified using online satellite imagery. https://doi.org/10.1371/journal.pone.0197445.t001 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 6 / 14 Table 1. Land use types, features included and excluded and spatial statistics. PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 6 / 14 Assessing urban carnivore density on lowest deviance values. All statistical analyses were computed in R 3.4.0 [40] and the GLMM was computed using the glmmPQL command in package MASS. ative fox group density (FGD) and badger group density (BGD) estimates and landscape composition in the 14 study sites. Relative fox group density (FGD) and badger group density (BGD) estimates and landscape composition in the 14 study sites. 1 i i id d l bli h d 1cities are considered long established 2 h l Relative fox group density (FGD) and badger group density (BGD) estimates and landscape composition in the 14 study sites. 1cities are considered long established d long established olonisation https://doi.org/10.1371/journal.pone.0197445.t002 Relative fox group density (FGD) and badger group density (BGD) estimates and landscape composition in the 14 s Discussion Judge et al, 2014 [34] state that social group abundance may be considered as useful as total population size (i.e. number of individuals) as it is less likely to vary at fine temporal scales, and is epidemiologically informative for disease transmission in social carnivores. Additionally, management actions are typically taken at group level. For both foxes and badgers, social group density estimates allow data to be compared historically and between different studies, whilst maintaining relevance to disease models [18]. Where an estimate of total population size is required, future studies additionally need to determine accurate current mean group size for their target species. In this study, we extrapolate from fox group density to individual (total pop- ulation) density by multiplying up with a group size of 3.4 (as used in [41]. Our field validation exercise yielded a comparable group size of 3.3. However, fox group size is documented to be highly variable, with up to 10 adults in a group in Bristol at peak population density [22]. Fur- thermore, a positive association between group density and group size cannot be assumed; for example, historical control operations resulted in a reduction in the number of individuals per group, but not the number of groups [36]. Likewise, long-term studies of rural badgers have shown temporal stability in social group density despite overall increases in population size [42] indicating that group size, rather than number of groups, was the unit of change. We used a group size of 5.5 to estimate individual density of badgers [35], however badger group sizes can also vary widely and are constantly in flux due to mortality and dispersal [31]. In this study, we recalculate fox group density in multiple cities in Britain for the first time since the 1980s. Our findings show similar group densities across towns/cities sampled between 2013 and 2015. However, we report marked increases since the 1987 study [36]. Our protocol is based on the same fundamental principles and therefore we have assumed direct comparability, although we recognise that both survey approaches may contain inherent errors and caution must be used when inferring differences/similarities. With this caveat in mind, we found the number of fox social groups in towns/cities with previously established fox popula- tions to be double that of Harris & Smith [36], and foxes to be present in all previously un-col- onised towns/cities. Results of fox cub records FSD FGD (km-2)(previous) Average FGD/city (km-2) (SE) BGD (km-2) Houses/fox family group London1 Croydon 1.04 77 0.168 0.515 1093 45 703.1 3.84 (0.7±0.4) 3.11 (1.02) 285 Surbiton 0.84 240 0.196 0.511 1614 232 2.39 675 Brighton1 Elm Grove 1.18 752 0.126 0.281 4266 129 37.8 3.39 (2.0±0.2) 3.53 (0.53) 3.70 1260 Hove 1.43 306 0.138 0.558 1155 120 3.49 2.67 331 Portslade 1.18 369 0.175 0.433 2149 137 4.25 0.87 506 Preston park 1.68 459 0.391 0.350 1104 256 2.98 370 Bournemouth1 0.92 366 0.085 0.536 1986 159 72.4 4.35 4.35 457 Portsmouth1 0.51 138 0.010 0.318 6028 21 13.2 3.91 (2.5±0.3) 3.91 1540 Newcastle2 Fenham 4.16 607 0.293 0.276 1907 38 51.2 1.68 (0) 3.24 (2.21) 1133 Heaton 0.83 395 0.244 0.389 1770 48 4.81 368 Huddersfield2 Almondbury 1.06 568 0.397 0.320 1530 337 14.6 4.73 (0) 4.15 (0.82) 324 Lockwood 0.84 858 0.262 0.196 2118 253 3.57 593 Norwich2 0.63 147 0.075 0.577 2087 19 36.6 4.73 (0) 4.73 441 Preston2 0.72 363 0.138 0.528 2177 8 4.0 2.76 (0) 2.76 789 Total 17.02 5645 1802 Mean 0.193 0.413 2213 3.63 (0.74) 3.72 2.41 648 SE 0.03 0.03 360 0.25 (0.40) 0.24 0.83 394 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 7 / 14 Assessing urban carnivore density recently colonised cities were included, the average FGD overall was five times that of previous estimates. If average FGD across cities (3.72) was multiplied by total suburban land cover this would equate to 33,092 [± 2,279 SE] fox social groups in suburban England, and a mean of one fox group per 648 houses across our study areas. Of the 6000 questionnaires distributed across three sites in Brighton, 1426 respondents reported 167 badger setts or sightings. Field studies verified and added locations and types of setts in the surrounding area to add to the distribution maps prior to analysis. This resulted in a mean BGD of 2.41 [±0.83 SE] and a mean individual density of 13.3 [± 4.56 SE], using 5.5 as group size (Table 2). If this value was representative of urban badger density, multiplication by suburban land cover would give an estimate of 21,971 [± 7,567 SE] badger social groups in sub- urban areas in England. Fox sightings data from Scott et al. (2014) were not correlated with FGD (Spearman’s rho, rs = 0.048, df = 7, P = 0.935). Results None of the landscape variables nor time since colonisation (COLON) predicted FGD (GLMM, full model deviance divided by null model deviance = 6.3%, t always < 0.9, P always > 0.05) and CITY accounted for only 0.003% of the residual variance (0.0001/3.79). PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 Assessing urban carnivore density used to develop rabies control models (maximum density 4 groups km-2 [11]). It would there- fore be advisable to revisit these models with the view that fox densities are now likely to be higher in many cities. As the method was based on fundamental principle of independent natal dens, FGD is also comparable with other countries, for example FGD in Melbourne, Aus- tralia, was estimated to be between 0.47and 2.55 km-2 [12]. Behavioural adaptation to exploit urban habitats, combined with natal habitat preference [43], suggest that foxes born and raised in urban areas are more likely to remain there, contrib- uting to the maintenance and expansion of populations [44]. Once colonisation has occurred, population growth will be determined by resources available e.g. food, rest and den sites [22] and by mortality and emigration. Foxes use a wide variety of features for den and rest sites [13] so these are unlikely to be density- limiting factors. Increases in the amount and suitability of suburban habitats, combined with increases in wildlife feeding by householders [8] may have supported population growth once colonisation occurred. There has also been a move away from lethal control and foxes have adapted their behaviour to reduce the risk of road mortality, although road traffic accidents remain a common cause of death [45]. Social group density is therefore likely to be affected by inter-group competition for resources, and overall density driven by social group size, which is itself limited by intra-group competition. We found no relationship between urban fox group densities and landscape composition. Furthermore, densities were very similar across sites in Brighton despite these areas contrast- ing in housing density and sociological composition. This indicates that previous studies using sociological data to predict fox densities [36] are no longer likely to be valid in modern subur- ban areas. Anthropogenic food sources, which can make up a considerable proportion of urban fox diet [46], have changed in availability, with increases in the provision by residents [8, 47]. These resources are typically provided in residential gardens, and their presence may reduce the influence of natural resources on fox density. The dynamics of such potential inter- relationships and associated behavioural mechanisms are unknown, and merit further study. The documented spread in distribution and potential increase in abundance of foxes in urban areas has also been reported in Europe (e.g., [48]) and globally (e.g., [12]). Discussion As fox density varies within and between cities, we cannot assume that this magnitude of change has occurred across Britain, although an overall increase appears plausible given the increased number of urban areas with foxes [24]. We found social group densities to range from 1.68 to 4.81 km-2, the higher limit of which exceeds that previously PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 8 / 14 It is still unclear why this phenomenon is occurring but suggestions include an increase in availability of habitats and resources that foxes can behaviourally adapt to and exploit [48], including sub- urban areas, which are expanding to provide housing for the growing human population. Mean density of badger main setts has been estimated nationally for different rural land use classes in England and Wales to give an overall density estimate of 0.485km-2 [34]. Our badger estimate from Brighton is five times greater than this, consistent with Davison et al (2008) who assert that urban sett counts are typically higher than rural ones. However, our replicates showed low precision, again in agreement with Davison et al. (2008). The habitat characteris- tics required by badgers (cover and sloping terrain of a suitable substrate for digging) are patchily distributed in urban areas hence urban badgers occupy non-contiguous home ranges and occur at variable social group densities [35]. Wider extrapolations for urban land classes are therefore untenable without further extensive study across multiple cities. Our density esti- mate of 2.41 km-2 is not as high as the 4.30 km-2 previously determined for one area of the same city (Brighton), but approximates to estimates from other urban areas (Hastings 2.05 km-2 and Bristol 1.88 km-2) (data from [35]). Further urban badger social group density esti- mation is required to correlate urban landscape structure with badger population size. Our method gives a viable approach to determining badger group density within suburban habitats where data are currently sparse. Our method allows estimation of relative fox group densities within and between cities, as validated by similar results from intensive surveys at one site. However, further calibration against a baseline estimate is recommended, at a range of different sites and cities. As the PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 9 / 14 Assessing urban carnivore density home range size and territorial spacing of both species varies a sensitivity analysis of the inte- gration method is recommended; i.e. calculation of robustness at different densities. The home range estimates in Brighton (see [39]) are small compared to previous studies, and ranges may be larger elsewhere, with fox cubs potentially moving between den locations that are greater than 200m apart. In such a scenario, the 200m integration method would lead to double count- ing and over-estimation of density. Fox social groups may have multiple litters in one year, which, if fragmented to smaller groups early, may also result in over-estimation. Likewise, bad- ger group inter-sett distances might be less than, or exceed, the 150m spacing used here. Therefore, sensitivity analyses are also required for integration distance. The resident response rate of 19% provided sufficient records of both fox and badger sight- ings to allow aggregation of sightings into centroids within GIS analysis, as representative of social groups. All sites provided sufficient data for estimates, demonstrating the ability to uti- lise records from residents across a country to determine numbers of social groups of both urban carnivores. However, response rate was variable, with questionnaire recipients from some sites being unresponsive. A low response rate in an area of low carnivore density would be at risk of insufficient returns, therefore incentives for response might be necessary. Responses via one route only (e.g. postal returns) should be monitored and a secondary route (e.g. door to door) followed up if response rate is low. Efficiency may also be increased by tai- loring delivery effort to housing density such that the same effort per unit area is deployed across all areas regardless of number of dwellings therein. The resident response rate of 19% provided sufficient records of both fox and badger sight- ings to allow aggregation of sightings into centroids within GIS analysis, as representative of social groups. All sites provided sufficient data for estimates, demonstrating the ability to uti- lise records from residents across a country to determine numbers of social groups of both urban carnivores. However, response rate was variable, with questionnaire recipients from some sites being unresponsive. A low response rate in an area of low carnivore density would be at risk of insufficient returns, therefore incentives for response might be necessary. Responses via one route only (e.g. postal returns) should be monitored and a secondary route (e.g. door to door) followed up if response rate is low. Efficiency may also be increased by tai- loring delivery effort to housing density such that the same effort per unit area is deployed across all areas regardless of number of dwellings therein. The method presented here could be adapted to compare densities in different cities with residential suburban areas across the globe and be used as a baseline to study spatiotemporal changes in fox and badger densities where this is desired. For example, foxes are colonising cit- ies in other parts of Europe where there are concerns of zoonotic disease transmission, includ- ing rabies and Echinococcus spp. [49, 50] and simple, efficient, and low-cost assessment of densities is paramount. Survey approaches are determined by ecological characteristics of the focal species, including social unit and group size, spatial distribution and space use, territori- ality and territory size. Table 3 shows a summary of key ecological traits of selected urban Table 3. Ecological traits of selected urban carnivores. Table 3. Ecological traits of selected urban carnivores. Red fox(Vulpes vulpes) Coyote(Canis latrans) Kit fox(Vulpes macrotis) European badger (Meles meles) Stone marten (Martes foina)c Striped skunk (Mephitis spp.) Raccoon (Procyon lotor) Bobcat (Lynx rufus) Distribution Northern hemisphere, Australia North America SW USA, NW Mexico Europe, Asia Europe, Central Asia North America North America North America Social group Yes Yes Yes Yes No No Males No Urban social group size 2.2–6.6 4–6 1–3 5.5 Solitary Solitary - Solitary Adult urban density 2–12 km-2 0.3–3 km-2 - 33 km-2 b 4.7–5.8 km-2 2–7 km-2 125–333 km-2 0.04–0.28 km-2 Exclusive territories Yes Yes Yes Yes Yes—Intersexual No No No Size of urban home range 0.14 km2 a 3–36 km2 1.2 km2 0.09 km2 (group) 1.13 (m)0.37 (f) km2 0.51–0.64 km2 0.05–0.79 km2 1.3–6.4 km2 Urban extent in home range Fully Mixed Fully Fully Fully Fully Fully Fully Seasonal breeder Yes Yes Yes Yes Yes Yes Yes Yes Typically one litter per social group Yes Yes Yes Yes No No No No A summary of ecological traits of selected urban carnivores. Information summarised from [2] with data from a [39], b [52] and c [53]; (-) denotes limited information. PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 10 / 14 Assessing urban carnivore density carnivores [2, 39, 51, 52]. To enable application of the method to other urban carnivores, such traits require consideration and the method adapted accordingly. Exclusive territories are essential for the method hence it is appropriate for coyotes (Canis latrans), kit foxes (Vulpes macrotis) and stone martens (Martes foina), but not for solitary species with overlapping terri- tories, such as skunks (Mephitis spp.) and raccoons (Procyon lotor). The clustering unit can be determined either by central den locations or litter locations, and mean territory size is required for buffer distance selection. Acknowledgments The project was funded by Earthwatch Institute, University of Brighton and APHA. We would like to thank all the volunteers who assisted with field work and data entry including R. New, N. Abery, S. Denis, J. Poole, K. Davies, S. Clare, C. Watson, A. Barbon, K. Cropper, E. Cropper, H. Bradley, H. Moorhouse, J. Crawley, T. Rogerson, Z. Keeble, A, Reynolds, A. Tarmey and Earthwatch Institute Volunteers. We would like to thank Philip J. Baker for comments on draft scripts and finally all the residents who responded to the questionnaire. For example, to estimate group density for coyotes, a large area would need to be surveyed due to extensive ranging. In this case a scatter approach might be engaged such that questionnaires cover a greater area with little or no additional effort. Ideally, the method would be employed in multi-species assessments, for example where kit foxes, coyotes and red foxes are sympatric (e.g. [53]) to maximise efficiency. Our finding of increased fox social group density relative to previous estimates has implications for ongoing disease surveillance and contingency plans, in particular relating to controlling poten- tial outbreaks of rabies and HAE. However, foxes are also considered potentially highly impor- tant in suppressing Lyme disease (Borrelia sp.) hosts in areas of human habitation [54], hence higher fox densities may be beneficial in controlling some zoonotic diseases. References 1. McKinney ML Effects of urbanization on species richness: A review of plants and animals. Urban Eco- systems, 2008; 11, 161–176. https://doi.org/10.1007/s11252-007-0045-4 2. Gehrt SD, Riley SPD, Cypher BL. Urban carnivores: ecology, conflict, and conservation. Johns Hopkins University Press, Baltimore; 2010. 3. Bateman PW, Fleming PA. Big city life: carnivores in urban environments. J Zool, 2012; 287, 1–23. https://doi.org/10.1111/j.1469-7998.2011.00887.x 4. 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Scott, Naomi Charman, Heidi Karlsson, Richard W. Yarnell, Aileen C. Mill, Bryony A. Tolhurst. Methodology: Dawn M. Scott, Rowenna Baker, Bryony A. Tolhurst. Project administration: Dawn M. Scott, Naomi Charman, Heidi Karlsson, Graham C. Smith. Resources: Dawn M. Scott. Supervision: Dawn M. Scott, Graham C. Smith, Bryony A. Tolhurst. Validation: Dawn M. Scott. Validation: Dawn M. Scott. Visualization: Dawn M. Scott, Rowenna Baker. Writing – original draft: Dawn M. Scott, Rowenna Baker, Bryony A. Tolhurst. Writing – review & editing: Dawn M. Scott, Richard W. Yarnell, Aileen C. Mill, Graham C. Smith, Bryony A. Tolhurst. 11 / 14 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 Assessing urban carnivore density PLOS ONE \| https://doi.org/10.1371/journal.pone.0197445 May 22, 2018 References Urban fox (Vulpes vulpes) population estimates and habitat requirements in sev- eral Brtisih cities. 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https://openalex.org/W3159285444	OpenAlex	Open Science	CC-By	2,020	Quasi-liquid crystals of electrons and positrons	Gianluigi Zangari del Balzo	English	Spoken	2,300	4,149	1. – Materials and methods Ultra fast uncooled photo-detector arrays based on HgCdTe (MCT) hetero-structures made by VIGO Systems S.A. [1] were used to perform real time bunch-by-bunch and turn-by-turn diagnostics at DAΦNE (Frascati) [2,3] and at Bessy 2, Helmholtz Zentrum, Berlin. The response of MCT photo-detectors in time (ns) was analyzed by a ﬁne-grain unconventional category statistical calculus system developed and patented (2011) by the present author. Given the limited space requirements, we refer amplius to [4-6]. 2. – Results The result of the analysis is a “bilobed” IR-SR emission proﬁle of each bunch, bearing the structure of a convolution of two peaks of which the second (hereinafter referred to as “delayed”) of smaller amplitude and delayed by a few hundred ps compared to the ﬁrst (hereinafter referred to as “main”), see ﬁg. 1. We used to represent this bilobed proﬁle as a ﬁnite sum of the atomic representation of a Banach algebra of Gaussian humps, a fascinating concept of Yves Meyer’s theory of ondelettes (wavelets). So, as a tribute to Yves Meyer, we used to call the phenomenon herein described as the “ondelette eﬀect” [7] . This choice expresses our methodological point of view and constraint, see [8]. Since the ondelette eﬀect was also observed in Berlin and Hefei (People’s Republic of China), we excluded that it was due to systematic errors (see also the analysis of time series, see [8]). Moreover, the velocity measurements at Frascati led to a small discrepancy in the value of the velocity of light, see [8]. Quasi-liquid crystals of electrons and positrons Gianluigi Zangari del Balzo(∗) Sapienza University of Rome - Rome, Italy Gianluigi Zangari del Balzo(∗) Sapienza University of Rome - Rome, Italy received 13 January 2021 received 13 January 2021 Summary. — A new statistical analysis of infrared synchrotron radiation (IR-SR) emitted by electrons and positrons at DAΦNE Φ-Factory (Frascati) and Bessy 2 (Berlin) reveals inner degrees of freedom within bunches of relativistic particles. We thus propose a new methodological approach towards a theory that describes the passage from a phase of maximum symmetry to a condensed phase having the qualities of a nematic mesophase of positrons and electrons. DOI 10.1393/ncc/i2021-21156-8 IL NUOVO CIMENTO 44 C (2021) 156 DOI 10.1393/ncc/i2021-21156-8 IL NUOVO CIMENTO 44 C (2021) 156 Creative Commons Attribution 4.0 License (https://creativecommons.org/licenses/by/4.0) (∗) E-mail: gianluigi.zangaridelbalzo@gmail.com 3. – A nematic mesophase The ondelette eﬀect questions the validity of the “rigid bunch” hypothesis [9, 10]. Consider thus the distribution of N electrons in a circular machine where we have a 1 1 2 GIANLUIGI ZANGARI DEL BALZO Fig. 1. – Ondelettes. (a) shows the plots of the amplitude (V) as a function of time (ns) of the IR-SR emission of a complete ﬁlling of 100 bunches of DAΦNE electrons (Frascati) with the detail of the bilobed SR-IR proﬁle of each bunch and its atomic representation through a Banach algebra of Gaussian humps, a fascinating concept of Yves Meyer’s theory of ondelettes or wavelets, hereinafter referred to simply for brevity as “ondelettes” [7]. (b) shows the plots of the amplitude (V) as a function of time (ns) of the IR-SR emission of a complete ﬁlling of 100 bunches of DAΦNE positrons (Frascati) with the detail of the bilobed SR-IR proﬁle of each bunch and our ondelettes. (c) shows the plots of the amplitude (a.u.) as a function of time per channels (ns) of the IR-SR emission of a single bunch of BESSY2 electrons (Berlin) with the detail of the bilobed SR-IR proﬁle of each bunch and our ondelettes. Data acquisition for DAΦNE electrons and DAΦNE positrons was carried out at SINDAB beam line by the group of Bocci et al., see [2], while for BESSY2 electrons it was acquired directly by the present author at the IRIS beam line (HZ-Berlin), see [5]. All the graphs, diagrams, tables and analyses (statistics, linear and non-linear regressions (ondelettes), time series analysis, category analysis and so on), all the models, methods and theoretical hypotheses for both DAΦNE and BESSY2, were created and performed by the present author with an unconventional category ﬁne grain statistical calculus and analysis, divided into 9 steps. We refer amplius to [4-6]. Fig. 1. – Ondelettes. (a) shows the plots of the amplitude (V) as a function of time (ns) of the IR-SR emission of a complete ﬁlling of 100 bunches of DAΦNE electrons (Frascati) with the detail of the bilobed SR-IR proﬁle of each bunch and its atomic representation through a Banach algebra of Gaussian humps, a fascinating concept of Yves Meyer’s theory of ondelettes or wavelets, hereinafter referred to simply for brevity as “ondelettes” [7]. 3. – A nematic mesophase For θ = 0 the form factor g(λ) is precisely the square of the Fourier transform of the bunch distribution. To deﬁne dp/dλ I took into consideration the eﬀect of the vacuum chamber screen. The cut-oﬀwavelength is λ0 ≈2h h ρ, where h is the total height of the vacuum chamber and ρ the radius of curvature of the trajectory of the particle. Following our ondelettes (see ﬁg. 2), we choose our form factor g(λ) for Gaussian bunches. For θ = 0 we will have 2πσz √ ln(N) < λ < λ0 = 2h h ρ, where σz is the bunch length. Therefore, to have a CSR emission we will have “short” bunches with large cutting wavelengths. In the case of real machines, the CSR contribution can be observed in the frequency range of THz. The distribution made of “micro-bunches” of σz length will have a density given by n (σz) = α δ (σz −σα z ) →⟨n (σz)⟩. ( ) α ( z ) ⟨( )⟩ The range of scale variation for σz is given by Λ < σz < Rs, where Λ and Rs represent, respectively, the lower and upper cut-oﬀscales, both depending on the cut-oﬀwavelength λ0 . That is, Rs is the maximum size of the distribution of micro-bunches. It is identiﬁed by the length of the whole cluster intended as “super-particle”, as Hofmann’s “rigid bunch” model. For scales such that Λ < σz < Rs we have a distribution of micro-bunches that look like “spindles”, all contained in a “super-particle” which emits synchrotron radiation along the direction of the motion. These micro-bunches are all ordered along a particular direction speciﬁed by a unit vector nμ called “director”, aligned with the direction of motion, while their centers of mass are distributed randomly (see ﬁg. 2). We thus have a distribution of bunches of particles characterized by a break (only) in the rotational symmetry. This is a condensed state resulting from a spontaneous symmetry breaking from a maximum symmetry state (homogeneous and isotropic “ﬂuid” phase) to a condensed phase where the rotational symmetry is broken. For congruence with our ondelettes (see ﬁg. 1), the model that comes closest to this phenomenon is the nematic mesophase of a liquid crystal [14, 15]. 3. – A nematic mesophase (b) shows the plots of the amplitude (V) as a function of time (ns) of the IR-SR emission of a complete ﬁlling of 100 bunches of DAΦNE positrons (Frascati) with the detail of the bilobed SR-IR proﬁle of each bunch and our ondelettes. (c) shows the plots of the amplitude (a.u.) as a function of time per channels (ns) of the IR-SR emission of a single bunch of BESSY2 electrons (Berlin) with the detail of the bilobed SR-IR proﬁle of each bunch and our ondelettes. Data acquisition for DAΦNE electrons and DAΦNE positrons was carried out at SINDAB beam line by the group of Bocci et al., see [2], while for BESSY2 electrons it was acquired directly by the present author at the IRIS beam line (HZ-Berlin), see [5]. All the graphs, diagrams, tables and analyses (statistics, linear and non-linear regressions (ondelettes), time series analysis, category analysis and so on), all the models, methods and theoretical hypotheses for both DAΦNE and BESSY2, were created and performed by the present author with an unconventional category ﬁne grain statistical calculus and analysis, divided into 9 steps. We refer amplius to [4-6]. coherent radiation term (CSR) along with an incoherent radiation term (ISR) [11-13]. The spectrum of radiated power will be coherent radiation term (CSR) along with an incoherent radiation term (ISR) [11-13]. The spectrum of radiated power will be (1) dP dλ = dp dλ N (1 −g (λ)) + N 2g (λ) , (1) where λ is the wavelength of the radiation, p the power emitted by a single particle, N is the number of particles per bunch and g the so-called “CSR form factor”, given by the following equation: g (λ) = +∞ −∞n (z) e2πicos(θ)z/λdz 2 , where 0 ≤g ≤1, n(z) is the bunch normalized distribution and θ is the angle between the longitudinal direction QUASI-LIQUID CRYSTALS OF ELECTRONS AND POSITRONS 3 Fig. 2. – A nematic mesophase of “quasi-liquid crystals” of electrons and positrons. ti h f “ i li id t l ” f l t d i id l ” Fig. 2. – A nematic mesophase of “quasi-liquid crystals” of electrons and positrons. Fig. 2. – A nematic mesophase of “quasi-liquid crystals” of electrons and positrons. Fig. 2. – A nematic mesophase of “quasi-liquid crystals” of electrons and positrons. z and the observation point. 3. – A nematic mesophase Coherence is therefore the average density as a function of the bunch dimension σz in the scale range given by the above interval (Λ, Rs), as follows: (2) lim σz→∞⟨n (σz)⟩∼ lim σz→Rs ⟨n (σz)⟩∼σDH z , (2) with DH ≥DT , where DH is the Hausdorﬀdimension and DT the topological dimen- sion. Deﬁnitely, we have a nematic fractal mesophase of micro-spindles emitting IR-SR ondelettes along the common direction of motion (ﬁg. 2). We used to call these struc- tures “quasi-liquid crystals of electrons and positrons”. Now we wonder the cause of the ondelette eﬀect. Evidences tell us that when a high charge density or the number of electrons in a bunch exceeds a threshold value, microstructures spontaneously ap- 4 GIANLUIGI ZANGARI DEL BALZO pear in the bunch, see [16, 17]. Ondelettes analysis is also a fast qualitative method de visu to detect inner bunch structures. From ﬁg. 1 we have that the main Gaussian peak (“main”) corresponds to the ISR emission of the whole bunch as a “super-particle” (super-spindle), while the small Gaussian peak (“delayed”) corresponds to the coher- ent emission (CSR) of the inner micro-spindles (which represents the theory deviation from Hofmann’s “rigid bunch model”). Considering therefore the interpolating Gaussian Ψ(t; μ, σ2) in the ondelettes, where t represents the time abscissa (ns), we have that μISR < μCSR ∀t, ∀bunch and Ψ(μISR) > Ψ(μCSR) , ∀bunch. As μCSR is the average delay time (ns) of the internal micro-spindles, between the slowest and the fastest micro-spindle inside the super-spindle all aligned with the direction of motion (see ﬁg. 2). Similarly, we note the diﬀerence ΔΨin amplitude (V) between the two ondelettes taking into account that the ISR term in the power spectrum (eq. (1)) is proportional to N, while the CSR term is proportional to N 2. Recall that particle densities scale like fractals (eq. (2)). The density of the incoherent particles is therefore greater, as expected. Given the limited space, we reserve the right to continue in a future work. REFERENCES [1] Piotrowski J. and Piotrowski A., VIGO System SA, MCT Photo detectors, http://www.vigo.com.pl/ (2006). [2] Bocci A. et al., The Time Resolved Positron Light Emission (3+L) experiment: A novel diagnostic tool for the DAΦNE positron ring. LNF-09/15 (2009). [3] Guidi M. C. et al., J. Opt. Soc. Am. A, 22 (2005) 2810. [4] Zangari G., Quasi-liquid crystals of electrons and positrons, https://doi.org/10.5281/ zenodo.3987312 (2020). [5] Zangari G., Time domain test of a fast MCT photo-detector with synchrotron light at BESSY II IR facility (IRIS), http://dx.doi.org/10.13140/RG.2.2.11483.18721 (2011). [6] Zangari G., Analisi statistica di massima congruenza applicata all’emissione di radiazione di sincrotrone: il caso della Φ-Factory DAΦNE: quasi cristalli liquidi di elettroni e positroni (KDP-Amazon) 2020. [7] Meyer Y., Wavelets and Operators (Cambridge University Press) (1992). [8] Zangari G., Theor. Biol. Med. Model., 18 (2021) 1. [ ] ( ) [9] Hofmann A., The Physics of Synchrotron Radiation (Cambridge University Press) (2004). [10] Schwinger J., On radiation by electrons in a betatron (1945), in A Quantum Legacy: Seminal Papers of Julian Schwinger (World Scientiﬁc) (2000) p 307 [9] Hofmann A., The Physics of Synchrotron Radiation (Cambridge University Press) (2004). [10] Schwinger J., On radiation by electrons in a betatron (1945), in A Quantum Legacy: [9] Hofmann A., The Physics of Synchrotron Radiation (Cambridge University Press) (2004). [10] S J O di i b l i b (1945) i A Q L [9] Hofmann A., The Physics of Synchrotron Radiation (Cambridge University Press) (2004). [10] Schwinger J., On radiation by electrons in a betatron (1945), in A Quantum Legacy: Seminal Papers of Julian Schwinger (World Scientiﬁc) (2000), p. 307. [11] Sannibale F., Byrd J. M. et al., Stable CSR in storage rings: A model, in Beam Dynamics Newsletter n. 35 (International Committee for Future Accelerators) https://icfa-usa.jlab.org/archive/newsletter/icfa bd nl 35.pdf (2004). [12] Mayes E. and Hoffstaetter G. H., Coherent synchrotron radiation simulations for the Cornell energy recovery LINAC, in Proceedings of 1st International Particle Accelerator Conference IPAC2010, http://cds.cern.ch/record/1208994. [13] Abo-Bakr M. et al., Coeherent emission of synchrotron radiation and longitudinal instabilities, in Proceedings of the 2003 Particle Accelerator Conference (IEEE) 2003. ( ) [14] de Gennes P., Theory of liquid crystals, in Pierre-Gilles de Gennes’ Works Collected By The “College De France”, (College de France) 2009. ( ) [15] Chandrasekhar S., Liquid Crystals (Cambridge University Press), 1992. [16] Roussel E. et al., Phys. Rev. ST Accel. Beams, 17 (2014) 010701. REFERENCES [ ] ( ) [17] Steinmann J. L. et al., Phys. Rev. Accel. Beams, 21 (2018) 110705.	36,110
https://openalex.org/W4362597413	OpenAlex	Open Science	CC-By	2,023	Multiple factors causing alignment instability in childhood concomitant strabismus	Yanan Guo	English	Spoken	3,394	5,950	Multiple factors causing alignment instability in childhood concomitant strabismus National Center for Children’s Health, Capital Medical University Jingjing Jiang (  jiangjingjing1980@163.com ) National Center for Children’s Health, Capital Medical University Design: Prospective cohort study. Methods: In total, 198 children with concomitant strabismus (57 esotropia, 141 exotropia) underwent repeat prism and alternate cover tests (PACTs) by two orthoptists who were certified by the study, and sensory tests were all performed once. We defined the alignment measurement as stable if the absolute value of the measurement difference between two orthoptists was within 10 prism diopters (PD) at distance or at near, and unstable if the difference was 10 PD or greater at distance or at near. We analyzed the relationship between the measurement variability and sensory results, patient age, and angle of deviations. Results: The mean age of the esotropia and exotropia patients was 68.5 ± 26.3 months (range, 36–164 months) and 96.1 ± 33.6 months (range, 22–200 months), respectively, and there was a significant difference in suppression related alignment instability, both at distance (P= 0.005) and at near (P = 0.039). Anisometropia also showed a significant difference at distance (P = 0.033) for alignment instability, and there was no significant statistical effect of age on measurement variability in our study. Alignment instability is positively associated with the angle of deviation, especially at distance (P = 0.019 for exotropia, P= 0.002 for esotropia). Conclusion: Clinical characteristics are proven related to alignment instability, and suppression is an important factor for alignment instability in childhood concomitant strabismus. Nevertheless, it would be entirely reasonable to consider alignment instability in relation to other factors, such as anisometropia and a large angle of strabismus, when evaluating patient deviations. Trial registration: Chinese Clinical Trial Registry, ChiCTR2200062404, Registered 4 August 2022. Research Article License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Additional Declarations: No competing interests reported. Page 1/13 Page 1/13 Abstract Purpose:To investigate the potential factors related to alignment instability in childhood concomitant strabismus. Introduction Childhood concomitant strabismus is the most common strabismus disorder, affecting up to 5.41% of Chinese children,[1] and it is associated with a significant negative impact on quality of life and a deficit in binocular vision.[2] It is often initially managed by prescription glasses and occlusion therapy, but when children finish amblyopia therapy and undergo alternate fixation, extraocular muscle surgery is the next intervention therapy indicated. Early surgical intervention has been proven associated with better stereopsis for esotropia patients,[3] but the timing of intervention for exotropia patients remains unclear.[4] No matter the type of strabismus, surgeons should weigh the stability of the deviation over several visits and determine surgery timing.[5] In the current literature, repeated measurements of the deviation are derived from data through routine visits, but because there is usually a time interval between each measurement, such as weeks or months,[6, 7] different kinds of source errors may be introduced to the Page 2/13 data. First, the Krimsky test and the prism and alternate cover test (PACT) may introduce errors in different measurement methods. Second, spectacle correction in patients may bring about a change in deviation for the next visit [8]. Third, the examiner taking the second measurement likely reviewed the previous measurement data, which could affect the current measurement. In addition, the natural course of diseases also influences the measurement results, as only 57% of infantile esotropia patients may have measurement stability (within 10 prism diopters [PD]).[3] In a population-based cohort of pediatric patients with intermittent exotropia, more than half were expected to have an increase of 10 or more PD within 20 years of their diagnosis.[9] data. First, the Krimsky test and the prism and alternate cover te different measurement methods. Second, spectacle correction in deviation for the next visit [8]. Third, the examiner taking the seco previous measurement data, which could affect the current mea of diseases also influences the measurement results, as only 57 have measurement stability (within 10 prism diopters [PD]).[3] In patients with intermittent exotropia, more than half were expecte within 20 years of their diagnosis.[9] Therefore, the characteristics of concomitant strabismus when measuring stability are still unclear, so this study aimed to investigate potential factors related to alignment instability and to avoid measurement errors over multiple visits to achieve a more reliable assessment of children with concomitant strabismus and provide better surgical plans. Patients The study approval was obtained from the Institutional Review Board of Beijing Children’s Hospital (2022- E-126-Y), and all procedures and data collection were conducted in compliance with the tenets of the Declaration of Helsinki. Further, informed consent was obtained from all patients and from their parents/legal guardians A prospective analysis of horizontal concomitant strabismus patients (ages ranging from 24 months to less than 18 years) who visited our strabismus clinic between August 2022 and November 2021 was performed. The inclusion criteria were horizontal concomitant strabismus with a deviation of 15 PD or more for near or distance, normal ductions, both eyes could fix at target after amblyopia treatment, and the ability to sit and cooperate with the PACT and sensory test. The exclusion criteria were patients who had fully accommodative esotropia, vertical deviations, A- or V-pattern strabismus, dissociated vertical deviation (DVD), nystagmus, restriction strabismus, paralytic strabismus, or other non-concomitant strabismus. Significant developmental delays or any known neurological diseases were excluded. Angle measurements were all obtained using the PACT, with patients wearing habitual refractive correction if prescribed. The patient viewed a target at a distant fixation (3 meters) and near fixation (1/3 meters). Then, base-in (for exotropia) or base-out (for esotropia) prisms were placed over one eye and the dissociation continued, with gradually increasing prism strengths until the angle of deviation was overcorrected. Prism strength was then decreased to identify the largest prism strength at which the deviation was neutralized. The largest prism magnitude that neutralized point was recorded as the angle of deviation. Data collected also included age, sex, amblyopia therapy history, and refractive error, and stereopsis was evaluated by the Titmus Stereo Test and suppression by Bagolini’s striated glasses. Page 3/13 To reduce the influence of examiner errors, all alignment measurements were taken by two orthoptists who were certified by the study, and previous measurement(s) were not reviewed when repeating the assessment. However, the sensory tests were all only performed once. Statistical analysis The study uses both categorical and continuous variables for analysis. In a continuous measurement, the absolute value of the measurement difference between two orthoptists is computed separately for at distance and near. The larger the difference, the more unstable the alignment measurement. In a categorical measure, we defined the alignment measurement as stable if the absolute value of the measurement difference between two orthoptists was within 10 PD for at distance or near and unstable if the difference was 10 PD or more for at distance or near. Data were analyzed using the statistical software Stata (version 15.1). Pearson’s Chi-square test was used to compare non-continuous variables between groups, and regression analyses were performed to determine whether there was greater variability with age or greater angle of deviations. P-values of less than 0.05 were considered statistically significant. Patient demographics and clinical characteristics The clinical characteristics and demographics of the patients are presented in Table 1. In total, 198 patients with concomitant strabismus enrolled in this prospective study. Among them, 57 (28.8%) had esotropia and 141 (71.2%) had exotropia. The mean ages of the esotropia and exotropia patients were 68.5 ± 26.3 months (range, 36–164 months) and 96.1 ± 33.6 months (range, 22–200 months), respectively. Page 4/13 Page 4/13 Table 1 Demographics and clinical characteristics of concomitant strabismus in this cohort study Variable Esotropia Exotropia Mean age (months) 68.5 ± 26.3 21 ± 96.1 ± 33.6 Sex (n, %) Male 24 (12.1%) 73 (36.9%) Female 33 (16.7%) 68 (34.3%) Refractive status (n, %) Hyperopia 49 (24.8%) 33 (16.7%) Emmetropia 0 (0%) 19 (9.6%) Myopia 8 (4%) 89 (45%) Anisometropia, n, (%) 7 (3.54%) 22 (11.1%) Amblyopia therapy history (n, %) 23 (11.6%) 20 (10.1%) Reliable Stereopsis (n, %) 27 (13.6%) 130 (65.7%) Suppression (n, %) 44 (22.2%) 61 (30.8%) Mean deviation (PD) At near 22 ± 13.3 24.5 ± 11 At distance 20.4 ± 12.2 19.7 ± 10.9 Table 1 The Relationship Between Alignment Instability And Angle Of Deviation Overall, we find that alignment instability is positively associated with the angle of deviation, where Fig. 2 shows this relationship concerning at distance. We can see that a larger angle of deviation at distance results in significantly higher alignment instability. Among exotropia patients, a 1-PD deviation at distance results in a 0.115-PD difference in the measurement (P = 0.019). Among esotropia patients, the effect is even greater. As the deviation at distance increases by 1 PD, the difference in the measurement increases to 0.270 PD (P = 0.002). By comparison, the effect of the angle of deviation on alignment instability at near is smaller for exotropia patients (P = 0.140), but it still has a positive effect for esotropia patients (P = 0.001; Fig. 3). The Relationship Between Alignment Instability And Age The scatter plots in Fig. 1 display the relationship between patient age and alignment instability. As the OLS regression coefficients (b Esotropia, At near = -0.009, P = 0.805; b Esotropia, At distance = 0.032, P = 0.449; b Exotropia, At near = -0.028, P = 0.154, b Exotropia, At distance = -0.008, P = 0.626) show, patient age is not significantly associated with measurement variability. Potential Factors Related To Alignment Instability According to Pearson’s Chi-square test analysis, among all the clinical characteristics (Table 2), there were no statistically significant differences in age, sex, refractive status, stereopsis, amblyopia therapy history, and type of strabismus related to alignment instability. There was a significant difference in suppression related to alignment instability, both at distance (P = 0.005) and at near (P = 0.039), and anisometropia showed a significant difference at distance (P = 0.033) with alignment instability. Page 5/13 Page 5/13 Page 5/13 Table 2 Comparison of alignment instability according to characteristics Variable At distance p At near p Stable N (%) Unstable N (%) Stable N (%) Unstable N (%) Age < 72 months 54 (72) 21 (28) 0.432 40 (53.3) 35 (46.7) 0.129 ≥ 72 months 82 (66.7) 41 (33.3) 79 (64.2) 44 (35.8) Sex Female 67 (66.3) 34 (33.7) 0.467 55 (54.5) 46 (45.5) 0.098 Male 69 (71.1) 28 (28.9) 64 (66) 33 (34) Refractive status Emmetropia 12 (63.2) 7 (36.8) 0.726 10 (52.6) 9 (47.4) 0.082 Hyperopia 55 (67.1) 27 (32.9) 43 (52.4) 39 (47.6) Myopia 69 (71.1) 28 (28.9) 66 (68) 31 (32) Anisometropia Y 15 (51.7) 14 (48.3) 0.033 19 (65.5) 10 (34.5) 0.519 N 121 (71.6) 48 (28.4) 100 (59.2) 69 (40.8) Amblyopia therapy history Y 26 (60.5) 17 (39.5) 0.189 26 (60.5) 17 (39.5) 0.956 N 110 (71) 45 (29) 93 (60) 62 (40) Reliable stereopsis Y 110 (70.1) 47 (29.9) 0.414 94 (59.9) 63 (40.1) 0.898 N 26 (63.4) 15 (36 6) 25 (61) 16 (39) Table 2 Variable At distance p At near p Reliable good stereopsis (≤ 100 sec arc) Y 82 (71.9) 32 (28.1) 0.252 68 (59.7) 46 (40.3) 0.880 N 54 (64.3) 30 (35.7) 51 (60.7) 33 (39.3) Suppression Y 63 (60) 42 (40) 0.005 56 (53.3) 49 (46.7) 0.039 N 73 (78.5) 20 (21.5) 63 (67.7) 30 (32.3) Type of strabismus Esotropia 34 (59.7) 23 (40.3) 0.081 32 (56.1) 25 (43.9) 0.469 Exotropia 102 (72.3) 39 (27.7) 87 (61.7) 54 (38.3) Th R l ti hi B t Ali t I t bilit A d A At distance Discussion Page 7/13 Page 7/13 Previous cohort studies have measured several visits for patient deviations, which cannot prevent patients’ loss of follow-up.[5, 7] Our study included a test–retest measurement simultaneously to ensure the quality is relatively reliable. By analyzing different clinical characteristics in our study, we found suppression is an important factor related to alignment instability. Regardless of whether at a distance or near, suppression can affect the measurement results. This phenomenon can likely be explained by the hemiretina trigger mechanism, as Pratt described in 1983,[10] where suppression is developed during visual immaturity to overcome diplopia caused by strabismus, as the image of the fixation object always falls on the same half of the retina; if the deviation is overcorrected by prisms, the image of the fixation object crosses the vertical retinal midline and falls on the opposite half of the retina in the deviating eye, which is a new situation that immediately triggers diplopia or a new pattern of suppression. Thus, for patients with suppression, it is difficult to recognize diplopia immediately by increasing the strength of prisms, which causes a variable neutral point and leads to measurement instability.[10, 11] In addition, patients with anisometropia show unstable measurements at distance, and we believe it has a strong relationship with suppression. In our study, we did not find statistical differences in patients with or without stereopsis, but we did observe patients with stereopsis or good stereopsis (≤ 100-sec arc), and the stability of measurements is generally higher. It can be inferred from our study that patients with better binocular vision and without suppression can obtain reliable measurement results. There is a stereotype of young children having poor coordination, and measurement stability must be worse. A 2008 Pediatric Eye Disease Investigator Group (PEDIG) study reported that unstable ocular alignment is common in childhood esotropia, especially in the youngest children with infantile esotropia (IET); for a difference in instability of ≥ 15 PD between any two measurements over four visits in an 18- week observation period, they found that 46% of children with IET were classified as having unstable alignment[7]. Birch et al [5] reported similar results, where only 28% of IET patients maintained a stable angle of deviation across four visits at approximately six-week intervals. Discussion In our study, there is no effect of age on measurement stability, a reasonable explanation is that we did not use the Krimsky method, which has been shown to lead to an overestimation of the deviation angle in young children; thus, we enrolled subjects all older than 2 years old who could cooperate with the PACT. Furthermore, our data also showed a tendency for an increased difference of deviation in patients with a larger angle of strabismus, regardless of whether at distance or near. The explanation for this finding is that the increments of the prisms used were 1 PD to 5 PD for a small angle of strabismus, but the increments for a large angle of strabismus could be from 5 PD to 10 PD, easily surpassing the 10-PD threshold. Hatt et al [12] evaluated 26 patients with intermittent exotropia with the PACT, and they also found a greater effect of the angle of deviation on variability. There are limitations to our study. First, we evaluated deviation twice, but only once for sensory tests, which should be considered in interpreting our results. Second, all patients enrolled cooperated with the PACT, ignoring the influence of many variable factors brought on by age. Page 8/13 Page 8/13 In conclusion, our data show that suppression is an important factor in alignment instability in childhood concomitant strabismus. Nevertheless, it would be entirely reasonable for other factors, such as anisometropia and a large angle of strabismus, to be considered when evaluating deviation in patients. In conclusion, our data show that suppression is an important factor in alignment instability in childhood concomitant strabismus. Nevertheless, it would be entirely reasonable for other factors, such as anisometropia and a large angle of strabismus, to be considered when evaluating deviation in patients. Availability of data and materials: All data generated or analyzed during this study are included in this published article. Funding: This research was funded by the National Natural Science Foundation of China (No. 81970844) and R&D Program of Beijing Municipal Education Commission (No. KZ202110025039). Author Contributions: Yanan Guo and Yuehan Guan collected data. Jingjing Jiang wrote and Li Li revised the manuscript. All authors have read and agreed to the published version of the manuscript. Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. Acknowledgments: I would like to express my gratitude to all those who helped me during this research. Acknowledgments: I would like to express my gratitude to all those who helped me during this research. Declarations Ethics approval and consent to participate: The study approval was obtained from the Institutional Review Board of Beijing Children’s Hospital (2022-E-126-Y), and all procedures and data collection were conducted in compliance with the tenets of the Declaration of Helsinki. Further, informed consent was obtained from all patients and from their parents/legal guardians. Consent for publication: Not applicable. Consent for publication: Not applicable. Consent for publication: Not applicable. Availability of data and materials: All data generated or analyzed during this study are included in this published article. References 1. Wang Y, Zhao A, Zhang X, Huang D, Zhu H, Sun Q, Yu J, Chen J, Zhao X, Li R, et al. Prevalence of strabismus among preschool children in eastern China and comparison at a 5-year interval: a population-based cross-sectional study. BMJ Open. 2021;11(10):e055112. 2. Wen G, McKean-Cowdin R, Varma R, Tarczy-Hornoch K, Cotter SA, Borchert M, Azen S. Multi-ethnic Pediatric Eye Disease Study G: General health-related quality of life in preschool children with strabismus or amblyopia. Ophthalmology. 2011;118(3):574–80. 3. Wong AM. Timing of surgery for infantile esotropia: sensory and motor outcomes. Can J Ophthalmol. 2008;43(6):643–51. 3. Wong AM. Timing of surgery for infantile esotropia: sensory and motor outcomes. Can J Ophthalmol. 2008;43(6):643–51. 4. Lavrich JB. Intermittent exotropia: continued controversies and current management. Curr Opin Ophthalmol. 2015;26(5):375–81. 4. Lavrich JB. Intermittent exotropia: continued controversies and current management. Curr Opin Ophthalmol. 2015;26(5):375–81. 5. Birch EE, Felius J, Stager DR, Sr., Weakley DR Jr, Bosworth RG. Pre-operative stability of infantile esotropia and post-operative outcome. Am J Ophthalmol. 2004;138(6):1003–9. 5. Birch EE, Felius J, Stager DR, Sr., Weakley DR Jr, Bosworth RG. Pre-operative stability of infantile esotropia and post-operative outcome. Am J Ophthalmol. 2004;138(6):1003–9. Page 9/13 Page 9/13 6. Hong S, Ha S, Seo Y, Kim S. Analysis of Postoperative Outcomes in Patients with Intermittent Exotropia According to Preoperative Change of Angle of Deviation. Korean J Ophthalmol. 2021;35(2):130–5. 7. Investigator PEyeD, Christiansen G, Chandler SP, Holmes DL, Arnold JM, Birch RW, Dagi E, Hoover LR, Klimek DL, Melia DL. Instability of ocular alignment in childhood esotropia. Ophthalmology. 2008;115(12):2266–2274e2264. 8. Chung SA, Kim IS, Kim WK, Lee JB. Changes in exodeviation following hyperopic correction in patients with intermittent exotropia. J Pediatr Ophthalmol Strabismus. 2011;48(5):278–84. 9. Nusz KJ, Mohney BG, Diehl NN. The course of intermittent exotropia in a population-based cohort. Ophthalmology. 2006;113(7):1154–8. 10. Pratt-Johnson JA, Tillson G, Pop A. Suppression in strabismus and the hemiretinal trigger mechanism. Arch Ophthalmol. 1983;101(2):218–24. 11. Pratt-Johnson JA, Tillson G. Suppression in strabismus–an update. Br J Ophthalmol. 1984;68(3):174–8. 12. Hatt SR, Leske DA, Liebermann L, Mohney BG, Holmes JM. Variability of angle of deviation measurements in children with intermittent exotropia. J AAPOS. 2012;16(2):120–4. Figures Page 10/13 Figure 1 The relationship between the difference in deviation and age (A, Esotropia, at near; B, Esotropia, at distance; C, Exotropia, at near; D Exotropia, at distance). The fitted lines are estimated from OLS regression models. Figure 1 Figure 1 The relationship between the difference in deviation and age (A, Esotropia, at near; B, Esotropia, at distance; C, Exotropia, at near; D Exotropia, at distance). The fitted lines are estimated from OLS regression models. Page 11/13 Figure 2 The relationship between the difference in deviation and mean deviation at distance (A, Exotropia; B, Esotropia). The fitted lines are estimated from OLS regression models. Figure 2 The relationship between the difference in deviation and mean deviation at distance (A, Exotropia; B, Esotropia). The fitted lines are estimated from OLS regression models. Page 12/13 Page 12/13 Figure 3 The relationship between the difference in the deviation and the mean deviation at near (A Exotropia; B Esotropia). The fitted lines are estimated from OLS regression models. Figure 3 The relationship between the difference in the deviation and the mean deviation at near (A Exotropia; B Esotropia). The fitted lines are estimated from OLS regression models. Page 13/13	30,424
W2607256975.txt_3	German-Science-Pile	Open Science	Various open science	null	None	None	Japanese	Spoken	5,900	17,488	17ニ於テハ下痢アリタルニ拘ハラズ尚ホ尿量ハ對照ニ比シ増加セルハ注意スベキ點ナリ.Pyrifer家痢,食慾不振於テハカカル副作用殆ドナク又比較的充分ナル發熱ヲ見ル事ヲ得タ.Py‑ 兎ニ於テハ尿比重甚シク増シタルニ對シSulfurolニ對照ニ比シ多少増シタルノ感アリ.皮 Pyrifer, Sulfurol家於テハ膚感受性ハ兎共ニ之等注射ニヨリ變化シ感受性ハ増シ或ハ減ジ一ツトシテ變化セザルハナテハNaハ Sulfurol注於テ其ノ度強キガ如シ. シSulfurolニカリシ.Pyrifer, 括家兎ニ於テモPyrifer, 兎體重ハ其ノ對照ニ比シ何レモ強度對スル當量比ヲ見ルニ輕度ノ減退アルモ,Kニク,Mgノ rifer, Sulfurol家多少増加ス. 總當價量強ク減退セリ. 次ニ皮膚ニ於ケルCaニ夫兎皮膚及ビ血液内電解質當量價爲ニ強輕度ノ減退アリ.Mgハ規 Kノ兎皮膚内電解質ニ就共ニ減退シ,Pyriferニ於テ特ニ強シ. 減退ハ甚ダ強ク又Pyriferニ於ケル其ノ度ハ Sulfurolニ ferニ Sulfurol家於ケルヨリモ遙カニ強シ.CaハPyri‑ 於テ甚ダ強ク減退シ,Sulfurolニ於テモ多少減退セリ.MgハPyriferニ於ケル輕度ノ減退, Sulfurolノニ其ノ總當價量Py‑ 強キ減退アリ.故 166 皮膚電解質ト皮膚感受性トノ關係ニ就テ(第1報) rifer, Sulfurol家兎共ニ減退シ,Pyriferニ其ノ度甚ダ強シ.血減退アリ.KハPyriferニフ.又兩氏ハ牝牛ノ乳ヲ搾ルト皮膚ノCaガスルノヲ見タ.故中電解質ニ就テハNaハPy‑ riferニ於テ輕度ノ減退,Sulfurolニ Sulfurolニ於テハ於テハ強度ノ於テハ殆ド變化ナク, 167 ニ皮膚ニハCaノ半減蓄積作用アル事明カナリト云ヘリ.雖然斯クノ如キ哺乳ニヨル皮膚Ca量共ニ同程度ニ於テ強ク減退セリ.MgハPyrifer家兎ニ於牙中ノCa減退シ易キヲ理由トシ以テ骨ヤ齒牙ハ兎ニ於テハ輕度増加單ナルCaノ蓄積所ナリト云ハントスル類ニ非ザケル減退アルモ,Sulfurol家セリ.從ツテ總當價量ハ共ニ減退シPyriferニテハ輕度ナルモ,Sulfurolニ於於テハ強度ナリ.即ハCaノノ減退ガ皮慮ニ何等影響ナク單ニ皮膚於テ多少ノ減退アリ.Caバ蓄積所ト考フルハ蓋シ哺乳ニヨリ骨ヤ齒ルカ.骨齒ノCaノ減退ニヨリ軟化スル事ハ直チニ夫レ自身ノ作用ヲ傷害スルモノナルヤ明カナルチ發熱劑ハ一般ニ皮膚及ビ血中電解質量ヲ減ゼシガ故ニ,皮メPyrifer注射ハ特ニ皮内電解質ヲ減退セシメ, ル皮膚生活作用ノ傷害ヲ推測シ得可シ. Sulfurol注射ハ特ニ血中電解質量ヲ減退セシムルモノナリ.皮膚含水量ニ就テモ兩者共甚シク減少セシムルモノナリ. 皮膚ノCa:KヲルニPyriferニ虜ニ於テモ亦カカルCa減家兎皮膚内電解質ハ健常家兎ニ於テモ動搖甚シキ事ハ屡々述ベタル處ニシテ,今横尾,齋藤,Brown氏見ルニ對照ニ於テハ1:2.03ナ於テハ1:1.95, Sulfurolニ於テハ 1:2.00ニシテPyriferニ Ca:Kト感受性トノ間ニハ終ニ一定シタル關係ヲ於ケル減退ヲ認ムルモ, 見出シ得ズ. 尚ホ著者ハ利尿劑ト發熱劑トノ關係ヲ知ランガ Brown氏ノ10匹ノ家兎ニ於テハ210%ニ搖最モ強ク,次シテ,齋藤氏ノ118%最 Brown氏ノ184%最モ強ク,次 %,次ニ著者ノ137%,齋横尾氏ノ153%ニ見ル事ヲ得ザリシハ殘念ナリ. 氏ノ306%,次 Rothman, Schaaf1), Dorffel氏 Schaaf兩シテ最モ強ク,次ノ次ハ著者ノ116%ナ最モ弱シ.MgハBrown ニ著者ノ214%,次小ハ齋藤氏ノ118%ナニ横尾氏ノリ. 等,其ノ他多ク氏ニ至リテハ緒言ニモ述ベ最モ少ナキモ實驗家兎數モ3匹 6匹,Brown氏ハ10匹,著 Surace37)氏ニシテ,横者ハ8匹ハ血液中ノCa量尾氏ハナリ. ハ變化少ナシト云タルガ如ク皮膚中ノ大部分ノ無機物ハ主ニ蓄積作ヘルモ,著者ノ實驗成績ニ據レバ健常家兎8匹用ノミニシテ皮膚生活作用トハ何等關係ナシト云血液Ca量ノ動搖範圍ハ12.32mgヨフガ如キハ著者實驗結果ニヨル健常家兎皮膚中ノニシテ甚ダ大ナリ.之總當價量ガ却ツテ血液中ノソレヨリ少ナキ點ヨリ圍ハ齋藤,Brown,著考フルモ當ラザルヲ知ル可シ.Laughlin, Mg中氏等ハ小牛,牡牛ノ皮膚内Ca量リ遙カニ多イ.牡 Theis ハ牡牛ノソレヨ牛ハ牝牛ヨリCaガ多イ故ニ兩氏ハ年齡ノ差ヨリハ寧ロ性ニヨル差異ナリト云 167 ニ上記ノ如ク齋藤氏ノ實驗結果ハ總テヲ通ジ動搖按ノ學者ガ皮膚ハ無機物ノ蓄積作用ヲナスト云ヒ, Rothman, ニ横尾氏ノ166 藤氏ハ112%ニシテ,其リ,同樣齋藤氏ノ108%ハ考ニ著者ノモ弱シ.Kモノ169%最注射後ノ皮膚感受性及ビ皮膚及ビ血液内電解質ヲ 125%,最シテ動ニ横尾氏ノ168%,次 165%ニ獸ハ總テ2日第6章ニ示セル等ノ健常家兎皮膚内電解質爲進ンデ家兎ニ利尿劑及ビ發熱劑ヲ注射セルニ試者ノ知ラント欲スル第6表ノ動搖範圍ヲ著者實驗ノ夫レト比較スルニ,Naハモ弱シ.CaモBrown氏後ニ死亡シ.著退ニ歸因ス Pick,酒者ノ例ヲ見ルモNa, 田氏ハClノ兎ニDiuretinヲ泄サレ,持リ5.862mg ニ反シ皮膚内Caノ最モ小ナル事ハ第6表ノ動搖範 K, Ca, ヲ見テモ明カナリ. 少ナイ食餌ニテ飼ヘル家與フレバ水分ノミナラズClモ續的ニDiuretinヲ排與フレバ先ヅ第1 168 西ニ皮膚中ノCl量ハ42%モ減少スルト.大田氏モ同樣無食鹽食再飼養家兎ニNovasurol, ヲ注射セシニNovasurolニ ganニルモ,血液中ノClニザリシト.今 Na量於テ265.56, 量ノ増加ヲ發見シ,Novasurolハ血球及ビ組織細胞中ノKニ膚中ノハ比シ明カナル Novasurolデニシテ對照ノソレハ267.9ナリ.即ハ215.95 モ多少ノ減退ヲ示セリ.即實驗結果ガ酒田,大ハチ著者ノ兎ニ於テハ血中Naノ減退ヲモ發生セル點ハ一致セズ. ニDiuretinヲハ變化ナカリシモ,此家兎與ヘルト水分ハ不變ナルモClハ第ニ減退スルト,即デ減ズル場合,夫血中ニ水分ヲ吸引シ利尿作用ヲ起スト.著者實驗結果ヲ見ルニ血中K量 surol 165.7,對血中K量ハSalyrgan 照173.1, 182.9, Nova‑ Salyrgan家兎ニ於テハノ増加ヲ認メ得シモNovasurolニ於テハ逆ニ減退ヲ示セリ. ハ食鹽多キ食物ヲ動物ニ與ヘタ ticaヲ與ヘ食鹽ヲ動物體ヨリ出ス樣ニスルモ血液中ノCaニハ變化ヲ及ボサズト.著劑注射家兎血液内Ca量者實驗ノ利尿ハSalyrgan家兎ニ於テハ甚ダ輕度ナルモ増加シNovasurolニ於テハ強度ノ減退ヲ示シタリ. 於テモ64.15%ニナル減退アリ.而ハ209.9, シテSalyrgan, シテ尚ホ一層著明シテ皮膚Na量 NovasurolデハSalyrganデハ190.6,對 Novasurol共ニ明カナルNa量チ著者實驗結果ニヨレバSalyrgan, Novasurol注射ニヨリ家兎皮膚内含水量,皮ハ共ニ減退シ,皮兎ニ於テハ皮膚内Na量強度ニシテ,同時ニ血中Na量ノ度上昇ニ傾クモ,尿Cl排ト.余ノPyrifer家 Sulfurolニ膚ノ減退モノ減退ヲモ招致セリ. 小林氏ニヨレバNovasurolノ量ノ増加,尿比重ノ増大田氏ハ無食鹽食餌飼養家兎ヲ「テルミン」,「チ Cl輕膚含水量ノ減退強度ナルNovasurol家重ノ減退,尿加等ノ確實ナル變化ヲ發見セリ. フスワクチン」ニヨリ發熱セシメタルニ血液中ノ照ハ253.1= 減退アリ.即内Na量シ Pyrifer, 兎皮膚中ノNa量於テハ214.03,對 Sulfurol發 242.1, ハ193.21, 照ハ253.1ニシテ, 熱家兎共ニ皮膚中ノNa量退ヲ認メタリ.而 rifer 出ハ證シ得ザリシシテ血液中ノNaニ Sulfurol 196.9,對照267.9ナテ兩發熱劑ノ場合共ニ減退シ,Snlfurol家減於テハPy‑ ルヲ以兎ニ於テハ其ノ減退亦甚ダ強度ナリ. 利尿作用ハ組織 Schroeder氏ハ硫黄ニ利尿作用アル事ヲ認メ, 168 皮膚電解質ト皮膚感受性トノ關係ニ就テ(第1報) Molitor氏ハ實驗動物ニ硫黄ヲヤリ尿素ヤ高張食ノ過敏性ヲ増シ,ソ鹽水ヲヤツタ時ト同樣滲透壓性利尿作用アル事ヲ K, 發見シダ.今 13例余ノ實驗結果ヲ見ルニ硫黄劑Sulfu‑ rol注射家兎1日照ガ」18ccナセリ.而平均ノ尿量ハ135.7ccニルニ比シ明カナル利尿作用ヲ表ハシテ比重ハSulfurolニニ,對照ニ於テハ1 .0145ニ Sulfurol家シテ謝 Ca, Mgハレニ添加スル「陰イオン」及ビ何等關係ナシト又Urbach氏ハノ天疱瘡患者ノ臨牀經驗ヨリシテ,天疱瘡患者ヨリ食鹽ヲ除ク時ハ病症惡化シ,食時ハ輕快ナル事ヲ認メ,此鹽ヲ與フル事實ヨリシテ天疱瘡患於テハ1.015ナル者ノ皮膚ニ食鹽ノ増加スルハ毒物的作用ヲナス蛋シテ反ツテ高シ.之ハ白質分解ヲ食鹽ガ防グ爲ノ保護作用ナリト云フ. 兎ニ於テハ尿中ニ電解質ノ排泄多キガ 1原因ナルモノノ如ク,皮 169 膚及ビ血液中ノ電解質斯ク考フル時ハKノ處デ云ヘル樣ニNaモ亦炎症ガ起ル場合,其ノ防禦作用トシテ其ノ部分ニ集ル總當價量ル甚シキ減退ハ之ヲ暗示セルガ如シ.即モノト考ヘ得.今余ノ例ニ於テ檢討スルニ健常家チSulfurol家兎ニ於テ皮膚感受性比較的強キ家兎Nr. 兎皮膚内總當價量ハ1122,血ノソレハ13.73ニ液内シテ對照ノソレハ各13 .35, 17.00 ナルニ比シ明カナル減退ヲ示セリ. Klose氏分,灰 Na量ニ據レバ病的條件ノモトニ於テハ水分,窒素等ノ含有量ハ無茶苦茶デ水分ハ多クナルモ少ナクナルモ總テノ疾病ノ場合Kガズ.疾減病ノ時ハ電解質ノ動搖強ク之ガ疾病ノ爲カ何ウカ不明ナリト.著者實驗ノPyrifer, 注射家兎ニ於テ皮膚及ビ血中Kノガ之ハKlose氏 Sulfurol 減退セルヲ見ルノ説ニ一致セルガ如シ.Haebler 氏ニ據レバ炎症ニヨル膿ヤ滲出物中ニハK甚ク,而モ炎症強キ程K量ト.Schade氏行ク程K増ニK多モ亦増加スルスト.Borelli氏モ皮膚内K量ハ炎症レニ由レバ皮膚キガ故ニ炎症ヲ起シ易イノデハナク皮膚感受性増セル家兎Nr. 比較的多シ.感 20ニ於テNr. 20ノ Nr. 18, 19ニ於テモ皮膚Kノ受性ヲ増セリ.以 furol家上ノ事實ヨリシテPyrifer, 兎ノ皮膚K量兎強キ減退ガ却ツテ感 Sul‑ ト皮膚感受性トノ關係ハ一定セザル事多キハ論ヲ俟タズ. Piepmeyer71)氏 169 ニヨレバNaノ熱家兎中 19ハ 15, 16, 多キモ,他ハ少ナ Mg増ニヨレバ70%ニ Brown6)氏 Na増多キセバ炎症セバ減ズト.尚ホKla‑ 於テCaノ加ガ皮膚感受性ヲ減ゼシメ,66%ニ増於テ僅カノK 増加ガ感受性ヲ増サシメタト.今余ノ實驗成績ニヨレバ健常家兎ノ比較的皮膚感受性強キNr 2ニ於テハNa, 比較的少ナシ.3ニ Mg比 . 2, 3, K比較的多ク, 於テハK僅カナルモ多較的少ナシ.6ニ於テハK比較的於テハNa,比較的多ク,Ca比較的少ナシ.7ニ多ク,Mg比較的少ナシ.Salyrgan, Novasurol 家兎ニ於テハ只一ツ感受性ノ増加セル家兎Nr. ヲ見ルニ他ニ比シNa, ナキモMgモニ於テハNa, 11 K甚ダ多ク,Caハ多少少多シ.Pyrifer, Sulfurol家兎ニ於テ感受性増シタ家兎Nr. 増加ノミガ細胞 11ハ 17, 18, 19中,17, uder5), ク,Ca, 感受性ヲ増ス事ヲ實證シタ.著者ノSulfurol家ノ1家兎Nr. 受性減ゼル家兎Nr. 準備状態高マリCa, ノナラン乎.即強キ減退ハ皮膚兎ニヨレバ皮膚ノK, Mgハ多キ皮膚ニ感受性高キ理 Novasurol家ニ感受性ノ増加セル家兎ハ皮膚Na量ハ此處ニ集リ來リテ未然ニ之ヲ防禦セントスルモチK量多キモ感受多シ.發ミ多少Na景 6ハ傾向アル事ハ云ヒ得. 6, 7ヲ見ルニNr. ルニ林氏ハ皮膚内KノニSalyrgan, 他ノ同樣實驗家兎ヨリモNa量ニ或種ノ炎症性障碍ガ先ヅ來ラントスル時ハ .K ナリ.然 1, 4, 5ハNa量ニ於テ皮膚感受性増加セル只1匹 Na量 2, 3, 6, 7 亦比較的多キモNr.3, 性比較的弱シ.次 Luithlen氏ハ〓瘡ニ於テハ炎症病竈ノ中央ヘニヨリ増加スル事ヲ發見セリ.是 7ハNa量少ナク,又Nr. シ.故ダ多ガ多イ.ソシテAcidose増加シ臨牀的ニ炎症増加スル程K量ニ於テ2, K他 17, 18, 19ヲ見ルニNr. ニ比シ比較的多ク,Mg比 17 較的 170 西少ナク,Nr. 18デハNa, 比較的少ナキモMgハ K比 Brugel, 組織中ノIonト組織中Kノ増シ,血 19デハNa比ニSulfurol家テハ大體ニ於テLuithlenノ Perutz, 較的少ナク,Caハ多シ.Nr. 的多ク,他ハ皆逆ナリ.故 Brack氏夫熱劑ハ感受性トCa:K:ト較兎ヲ除キシテ他ハ場合ハ一般ニCaニ對シテ比較的Kノ大ナル時,皮膚感受性高クナル等ニヨルト血液中ト Rothman28)氏液中ノCa増サバサバ組織中ノCa 變化ハ組織ノCa, Kノハ關係ヲ全ク破壞スルモノト考ヘラル,而モノノ如シ. 増シ,血液中ノK増 Caノ規説ニ一致スル事多シ. ハ丁度逆デ,血液中ノK, 川ハ血液ハ水分及ビ鹽類含有量ガ外界ノ状態ニヨリ影響サレニクイノハ,皮膚ヨリ不足ナル水分又ハ鹽類ヲ攝取シ,又餘レル水分又ハ鹽類ヲ皮膚ニ與ヘハレカラダト.酒田氏モ皮膚含變化ヲ表ハス鏡ノ樣ナモノダト.今著者ノ實驗例水量ニ就テ同樣ナ考ヲモツテ居ル.以ニ就テ調査セルモ終ニ一定セル關係ヲ發見シ得ザリ察スレバ血液中ノ電解質ハ甚ダ變化シ難ク,之リキ. 反シ皮膚中ノ電解質ハ甚ダ動搖シ安キガ如キ觀ヲ遠山,林兩氏ハCa:Kガ1:4カラ1:5ナ皮膚感受性モ正常ナルモ,何 1:3トナルモ,1:6ト等カノ原因ニヨリナルモ感受性亢進スルト.横尾氏ノ實驗結果ヨリCa:Kト係ヲ見ルニ遠山,林樣ダ.齋感受性變化トノ關兩氏ニ合致スル事ガ少シ多イシテ,健康家兎ニテモ感受性強キモノハCa:Kガ1:4.96,弱ナリト.又キモノハCa:Kガ1:3.58 リ甚シクK量上昇シ,感ガ各々1:3.46, 1:3.76, 少ナシ.Salyrgan, 1匹 :Kハ他ノ5匹 1:2.83ニノ家兎ヨリモ高 17, シテ其 Sulfurol家 18, 19ノ3匹 12, 13, 14, 15, 16, 17, 18, 19, 20ニ於テ減退セルモ血液中ハCaハ正常ナリ.Mgニ 18, 19, 20ニ於テ甚シキ減退アルニ血 13, 14, 於テ減退スルモ血液中ニ於テハチ利尿劑Salyrgan, rol注射家兎及ビ發熱劑Pyrifer, Novasu‑ Sulfurol注射家兎ニ於テハ皮膚電解質ニ甚シキ動搖ヲ來ス事アルモ血液内電解質ハ常ニ正常値ヲ失ハズ. 高イ.又比多少 Luithlen氏ノ實驗ノ結果ハ家兎ニ一方的食餌ヲ與フル事ニヨリ皮膚ニDemineralisationノ起驗家兎ニ於テ只ル事ヲ示シテヰル.又Nathan, 於バ飢餓動物ヤ渇水動物ノ皮膚内含水量ヤ皮膚内電テハ其ノCa キモ,其ノ値ハリ兎中感受性増セル家兎ハCa:Kハ1:2.16, 受性減ゼル家兎Nr. Salyrgan家兎皮膚含水量平均値ハ68.0ニ其ノ對照ノ平均値ガ73.8%ナ減少セリ.Novasurol家テ顯著ナル減退ヲ示セリPyrifer家リ.故ニ 66.75%ニ皮テハ66.10%ニニ於1ルソレハ1:2.72, Pyrifer, Sulfurolノ膚感受性トノ關係ハ全ク一定ノ規定ナク,即如キ發熱家兎ノCa:Kトチ發シテ, ルニ比スレバ明カニ 20 15, ニヨレノ實驗結果ヲ見ルニ 16, シテ,感 1:2.82ナ解質總量ハ減退スルト.余 Stern72)氏 1:1.82, 1:1.45ニ 1:1.24, 於テモシテNr. シテ健常家兎ノ平均値ナル1:3.38ヨ低シPyrifer, Nr. 11ニ液中ノK 9, 11, 9, ハCa:K 4ニ於テハCa:Kモノ感受性増セル家兎Nr. 甚シキ減退アルモ,血ハ尚ホ正常家兎ノ範園内ヲ出デズ.Caニ於テ皮膚中ノKノニ15, 液中ニ變化ナク,皮膚中ハ總當價量モNr. ノ健常家 1:2.97ニ Novasurol實 18, 19ニ減退ノ傾向ナシ.即除キテハ總テ平均値ヨリCa:Kガ較的感受性弱キ家兎Nr. 16, 17, 者 2, 3, 6, 7デ 1:4.21, 10, 11, 12, 13, 14, 20特 15, 16, 17, 18ニ平均1:3.38ニハ内多少感受性強キ家兎Nr. ハ今之等ニ就兎Nr. 受性減ゼル家兎ニ於テハ又甚シク低下スルト.著兎皮膚内電解質ノCa:Kハ果シテ事實ナリヤ,余テ余ガ實驗結果ヨリ總括考察スルニ,家於テモNr. 實驗ニヨリ感受性増セル家兎ハCa:K ハ1:4.32ヨ抱カシム.之 Nr. 藤氏ノ實驗ニヨル健常家兎ノ皮膚内Ca Kハ1:4.3ニ 7ヲル間ハ上ノ言句ヨ兎ニ於テハ64.15%ニシテ明カニ減少ス.Sulfurol家シテ同樣減退ヲ示セリ.皮解質總當價量ハ對照デハ13.35, シ兎ニ於テハ Salyrgan家兎ニ於膚内電兎デ 170 皮膚電解質ト皮膚感受性トノ關係ニ就テ(第1報) ハ11.74, Novasurol 11.22ニシテ,總 10.35, Pyrifer 9.94, Sulfurol 中ニ於テハNa, テノ實驗家兎ニ於テ相當多量ノ減退ヲ示セリ.故ニ利尿劑Salyrgan, 發熱劑Pyrifer, Sulfurol注含水量ノ減退 9) Novasurol, 射家兎ハ總テ著明ナル皮膚ノDemineralisationヲ示シ,同減退アリ,總時ニ皮膚 17.1 K, Mgノ輕度ハ減退,Caノ當價量亦甚シク減退ス. Salyrgan注 neralisationヲ射家兎ニ於テハ皮膚ノDemi‑ 起スモ血液中電解質ハ却ツテ増加セントスル傾向アルニ反シ,Novasurol注ニ於テハ皮膚ノDemineralisationノヲ示セリ. 強キ射家兎甚シキノミナラズ又血液ノ電解質總量モ減退セリ. 第7章結 10) 論 1) 健常家兎皮膚含水量ハ平均66.33%ナ 2) 健常家兎皮膚内電解質ハ平均(mg%)Na: 235.07, K:94.38, 13.76ニ Ca:14.59, Mg:4.84,總 Mg:5.34,總當價量:14.40ナ膚中ニ於テモ血液中ニ於テモNa及體等シク,皮量,皮膚中ノK量膚中ノCa量シテ,總ハ血液中ノCa量チ皮ハ大ノ約半ノ約2倍ニ電解質量ハ大體皮膚中モ血液中ニモ同量含マレ,寧 3) リ.即ハ血液中ノK量對スル他Ionノ當量比ハ 4) 16.5%, 5) 液ニ於テハ23.08:10.82:1:1.13ナ Ca:116/, Mg:214%ナ Novasurol注リ. 射家兎ハ確實ニ利 rganヨシテ比シ利尿作用甚シク優ノ副作用又Salyrganヨハ共ニ減退シ,其リ. 比重減退等ヲ生ズ.而リ強シ. ケルヨリ甚シク,血ハPyriferニ Salyrgan注 Ca, Mgハ於テハSulfurolニ於於テ於テハ減退セリ.皮 Sulfurol共ニ甚シキ減退於テ甚シク,血液中ノK, 於テハCaノ Ca ミ減退シ,Kニ於テハ於テハCaノ強度ノ減僅カノ減退アリ.皮膚内MgハPyrifer 於ケル甚シキ減退ヲ示セリ.血液中ニ於テハPyriferノMg ハ少シ減退セルニSulfurolニ於テハ多少増加セリ.皮 Sulfurol共膚總當價量ハPyrifer, ノ度ハNovasurolノ方ガSaly‑ 量ハPyriferニ於テ特ニ著明ナリニ減退血液中ノ總當價於テ輕度ニ減退シSulfurolニ於テハ明カニ減退セリ. 射ニヨリ其ノ家兎皮膚内電解輕度ノ減退,Kノ増加アリ. Caノ 14) Ca:KハPyrifer, ニ對スルKノ Sulfurol家輕度ノ減退アリ.Ca:Kト兎共ニCa 皮膚感受性トノ關係ハ一定ナラズ. 減少スルヲ認メタリ. Novasurol注 Mgノ兎皮膚内Naハニ於ケル僅カノ減退ニ比シSulfurolニシPyriferニ其ノ總當價量モ減退ス.血液中ニ於テハK, 増加,Mgノ兎共不液中ノNaハPyriferニ CaハPyrifer, 兎皮膚含水量リ強度ナリ. 質中Na, 射家兎ニ於テハ皮膚内ニ於テ甚シキ減退,Caノ相當ノ減退アリ, 爲ニ其ノ總當價量ノ甚シキ減少ヲ認メタリ.血液 171 ビSulfurol家共ニ減退セルモPyriferニ膚内K, Sulfurol家受性或ハ増加シ,或ハ減退セリ. Pyrifer及ビSalyrgan家 Novasurol及 K, 13) 退,Kノルモ,其 8) 規則ニ變化シ,感 K:137%, NovasurolハSalyrganニ Na, 皮膚感受性ハPyrifer, 全ク變化ナキニSulftvolニ尿起リ尿量増加,尿 7) 12) 健常家兎皮膚内電解質量ノ動搖範圍ハNa: Salyrgan, 6) 兎ニ於テハ反ツテ相當度ノ利アルモ特ニPyriferニ皮膚ニ於テハ平均Na:K:Ca:Mgハ14.0:3.38: 1:0.49,血兎ニ於テハ尿量甚シク減少セルハ變化殆ドナキモSulfurolニロ血液中ノソレノ方ガ多シ. 健常家兎ノCaニ Pyrifer家ニ反シSulfurol家リ. 尿作用ヲ表ハセリ. K:169.26 ビMg量射ハ皮膚感受性ヲ高メル事ア 11) 當價量: シテ血液中ノソレハNa:212.30, Ca:8.02, リ. Salyrgan注本稿ヲ終ルニ臨ミ本研究ヲ余ニ命ゼラレ終始御懇篤ナル御指導ト御校閲ヲ賜ハリシ恩師根岸教授ニ滿腔ハ謝意ヲ表ス. 西 172 川規文夫獻 1) Rothman u. Schaaf, Handb. d.H.u.G. v. Jadassohn, 1/2, 1929. 2) Dorffel, Arch. f.D. u. S., Bd. 174, Ht. 1, S. 1, 1936. 3) Luithlen, 35) Kramer-Tisdall-Denis 量法. 36) Wien. Klin. Wschr., S. 703, 1911; Arch. f. exp. 411頁. 37) Surace, Path., 68, 209, 1912. 4) Wiechowski, Munch. Med. Wschr., Nr. 34, S. 1082, 1921. 5) Klauder, 39) Nathan Arch. of. Dermat. a. Syphil., 11, 283, 1925; 15, 1, 1927; 19, 52, 1929. 6) Brown, J. of biol. 學的微量測定法;小 702. 松島不二,國 38) Borrell, 藤憲三著,醫化家醫學會次雜誌,第255號, Zbl. f.H.u.G., Bd. 42, S. Zbl. f.H.u.G., Bd. 43, S. 271. u. Stern, Zbl. f.H.u.G., 323, 1931. Dermat., 法;須金井良一著,生化學的微量定 40) Carrie 173, 606-611, 41) 1936. Chem., 68, 729, 1926; 75, 789, 1927. 7) Nathan u. Stern, Dermat. Z., 53, 451; 54, 14; 54, 232, ski, 1928. 8) Urbach, Klin. Wschr., 2, 17; 11, 2094- 43) Richter, 2096, 1929; Arch. f. Dermat. u. Syphil., 145, 220, 1924; 155, 1928; 156, 73, 1928; Biochem. Z., 196, 535, 1928. 9) Waterman, Biochem. Z., 1934. 44) Padtberg, 1910. 45) Baird u. Haldne, 259, 1922. 46) Abe u. Sakata, 1925. 立醫科大學雜誌,第9卷,昭 Munch. Med. Bd. 38, S. u. Heemeyer, 和8年. Wschr., Arch. f.D.u. Arch. f. 中島,京都府 42) Wiechow- Nr. 34, 1082, 1921. Syph., Arch. Bd. 170, S. 348, exp. Path., 63, 60, J. of Physiol., Arch. 56, f. exp. 133, 535, 1922. 10) Gans, Dermat. Wschr., 78, Path., 249, 1924. 11) Dahn, Dermat. Wschr., 82, 425, 1926. 12) Habler, Klin. Wschr., II, 1569-1572, 理及ビ療法. inn. Med., 1929. 13) Pinkussen, L., Biochem. Z., 182, 1927. f. inn. Ned., Bd. 6, 1923. 50) Feher, 14) Meltzer u. Auer, Amer. J. of Physiol., 15, 387, 1905-190; 15) Becku, Z. exp. Med., 67, Klin. Wschr., S. 964, 1929. 51) 市村丑雄,岡醫雜,舞513號. 52) 1929. 16) Kolm u. Pick, Pfluger's Arch., 185, 53) Block, Wien. Med. Wschr., 50, 1925. 54) Mauck, Arch. f. Dermat., 135, 208, 1921. 55) 235, 1920. 17) Zondek u. Aschheim, Klin. Wschr., Nr. 18, 831, 1928. 18) Klaus, Biochem. Z., 163, 41, 1925. 19) Ehrismann, Arch. f. exp. Path., 134, 1928. 20) Pincussen, Biochem. Z., 182, 359, 1927. 21) Klauder u. Brown, Arch. f. Dermat., 11, 283, 1925; 15, 1, 1927; 19, 52, 1929. 22) Bornstein, 山,林,皮 Biochem. Z., 172, 133, 泌誌,第26卷,第12號. 誌,第29卷,第12號. 25) 23) 遠 1926. 24) 横尾,皮泌齋藤,皮泌誌,第35卷, 第5號. 26) Engels, Arch. f. exp. Path., 51, 346, 1904. 27) Urbach, Arch. f. Dermat., 156, 73, 1928. 28) Rothman, Arch. f. Dermat., 131, 549, 1921. 29) Sakata, 1923. 30) Konigstein, 27, 799, 1926. f. exp. Wien. 105, 11, Klin. Wschr., 32) Schade, Z. exp. Path., 33) Wiechowski, Munch. Nr. 34, 1082, 1921. schr. f. gesamt. Path., Nr. 31) Rothstein, Berl. Klin. Wschr., Nr. 7, 154, 1920. 1, 1913. Arch. 34) Hahn 14, Med. Wschr., u. Taegr, Zeit- exp. Med., Bd. 76, S. 513, 1931. 105, 47) 佐々廉平,腎臟疾患ノ病 48) Saxl u. Heilig, Wien. Arch. f. Bd. 3, 1921. 49) Schur, Wien. 根岸,中西,岡 Arch. Wien. 醫雜,第557號. Nobl, Zbl. Hautkrkh., 11, 1924. 56) Kantor, Zbl. Hautkrkh., 11, 1924. 57) Gordonoff, Scheiz. Med. Wschr., Nr.46, 1926. 58) Schroew der, Kl. Wschr., Nr.46, 1927. 59) Meyer u. Bisch, Zeitschr. f. Kl. M., Ht. 4-6, 1922. 60) Maliva, Wien. Arch. f. inn. Med., 1924. 61) Spiro, Dtsch. 正雄,皮 m. W., Nr. 21, 1926. 62) 尼ケ崎泌誌,第44卷,第1號. 63) Gordonoff, Schweiz. med. W., Nr. 56, 1926. 64) Campanacci, Kl. W., 1926. 65) Molitor, Wien. Kl. W., 35, 40, 1927. 66) Schlesinger, Munch. m.W., Nr. 31, 1931. 67) Heubner, Zbl. Hautkrkh., 20, 1926; Biochem. Z., 122, 128, 1921. 68) Spiethoff, Dermat. Wschr., 1077, 1925. 69) Schwarz, Zbl. f.H.u.G., 第48年,第9號. 133-141. Bd. 48. 71) Piepmeyer, 72) Nathan u. Stern, 70) 大田,岡醫雜, Dermat. Z., 70, Zbl. f.H.u.G., Bd. 38, S. 193. 172 皮膚電解質ト皮膚感受性トノ關係ニ就テ(第1報) 173 Aus dem, Dermato-UrologischenKlinik der MedizinischenFakultat Okayama (Vorstand: Prof. Dr. H. Negishi) Experimentelle Studien uber lichkeit der die Haut Beziehungen and ihren zwischen der Empfind- die Empfind- Elektrolyten. (I. Mitteilung.) Uber den Einfluss von Diuretica und und Elektrolyte lichkeit die Fiebermitteln der auf Haut. Von Dr. Norio Eingegangen Nisikawa. am 26. September 1938. Der Verfasser untersuchte die Empfindlichkeit der Haut and dabei gleichzeitig auch die Elektrolyte in der Haut und im Blute nach Verabreichung von Dinretica (Salyrgan und Novasurol) und Fiebermitteln (Pyrifer und Sulfurol). Die Resultate sind folgendermassen; 1) Der Wassergehalt in der gesunden Kaninchenhaut ist durchschnittlich 66.33%. 2) Der normale Wert der 4 Elektrolyte der Haut (Na, K, Ca, Mg) beim mannlichen, erwachsenen, weissen Kaninchen betragt im Durchschnitt 235.07; 94.38; 14.59; 4.84 und das Gesamtaquivalent ist 13.76. Im Blute betragt derselbe Wert beim gleichen Kaninchen durchschnittlich 212.30; 169.26; 8.02; 5.34 und das Gesamtaquivalent ist 14.40. 3) Durch Injektion von Salyrgan und Novasurol bei den Versuchstieren trat Diurese auf, die Harnmenge vermehrte sich und das spezifische Gewicht des Harns nahm ab. 4) Sowohl bei Novasurol- als auch bei Salyrgan nahm beim Versuchstier der Wasser- gehalt der Haut ab und zwar bei Novasurol starker als bei Salyrgan. 5) Bei Salyrganinjektion verringerten sich Na, Ca und Mg in der Haut, aber K vermehrte sich, und zu gleieher Zeit trat im Blut eine Vermehrung von K und Ca sowie eine Abnahme von Mg ein. 6) Nach Novasurolverabreichung zeigte sich in der Haut eine deutliche Vermehrung von Na, K sowie Mg und eine mittelmassige Abnahme von Ca, dagegen im Blut eine geringe Abnahme von Na, K sowie Mg und eine deutliche Abnahme von Ca. 7) Bei Salyrganinjektion steigerte sich zuweilen die Empfindlichkeit der Haut, aber durch Novasurolinjektion blieb sie unbeeinflusst. 8) Bei Pyriferinjektion zeigte sich eine deutliche Abnahme der Harnmenge; die Sulfurolinjektion hatte eine massige diuretische Wirkung. 9) Die Empfindlichkeit der Haut zeigte naeh Injektion von Pyrifer oder Sulfurol keine bestimmten Veranderungen. 173 174 10) Der Mineralgehalt der Haut und im Blut wies bei Pyrifer- und Sulfurolinjektion folgende Veranderungen auf: Der Na-Gehalt in der Haut nahm bei Pyrifer- sowie Sulfurolinjektion ab, und zwar bei ersterer deutlicher als bei letzterer. Der Na-Gehalt im Blut zeigte bei Pyriferinjektion keine Abweichung, nahm aber bei Sulfurolinjektion ab. er K- und Ca-Gehalt D in der Haut nahm sowohl bei Pyrifer- ale auch Sulfurolinjektion auffallend ab, besonders stark war die Abnahme bei erstere. Was den K- und Ca-Gehalt im Blut angeht, so zeigte sick bei Pyrifer eine Abnahme von Ca, bei Sulfurol eine starkere Verringerung von Ca und eine schwachere von K. Per Mg-Gehalt in der Haut zeigte eine geringe Abnahme bei Pyriferinjektion eine starkere bei Sulfurol. und Der Mg-Gehalt im Blut verringerte sich ein wenig bei Pyrifer-, nahm aber auffallend zu bei Sulfurolinjektion. 11) Die Beziehung zwischen der Empfindlichkeit der Haut und dem Verhaltnis von Ca und K war nicht konstant. (Autoreferat) 174.	32,339
https://openalex.org/W3118281305	OpenAlex	Open Science	CC-By	2,021	Synthesis, crystal structure and photophysical properties of chlorido[2-(2′,6′-difluoro-2,3′-bipyridin-6-yl-κ<i>N</i> <sup>1</sup>)-6-(pyridin-2-yloxy-κ<i>N</i>)phenyl-κ<i>C</i> <sup>1</sup>]platinum(II)	Ki‐Min Park	English	Spoken	6,870	22,761	Ki-Min Park,a Cheol Joo Moon,a Sanghyun Paekb and Youngjin Kangc* Ki-Min Park,a Cheol Joo Moon,a Sanghyun Paekb and Youngjin Kangc* Received 4 January 2021 Accepted 5 January 2021 Edited by W. T. A. Harrison, University of Aberdeen, Scotland Keywords: crystal structure; 2,30-bipyridine; platinum compound; luminescence; OLED. CCDC reference: 2053861 Supporting information: this article has supporting information at journals.iucr.org/e Received 4 January 2021 Accepted 5 January 2021 aResearch Institute of Natural Science, Gyeongsang National University, Jinju 52828, Republic of Korea, bDepartment of Chemistry & Energy Engineering, Sangmyung University, Seoul 03016, Republic of Korea, and cDivision of Science Education, Kangwon National University, Chuncheon 24341, Republic of Korea. Correspondence e-mail: kangy@kangwon.ac.kr Edited by W. T. A. Harrison, University of Aberdeen, Scotland The title compound, [Pt(C21H12F2N3O)Cl], crystallizes with two crystallogra- phically independent molecules (A and B) in the asymmetric unit, which adopt similar conformations. The PtII atoms in both molecules adopt distorted square- planar geometries, coordinated by one C and two N atoms from the tridentate 20,60-diﬂuoro-6-[3-(pyridin-2-yloxy)phenyl]-2,30-bipyridine ligand and a chloride anion: the C and Cl atoms are trans. In the crystal, C—H Cl/F hydrogen bonds, F and weak – stacking interactions between adjacent A and B molecules and between pairs of inversion-related B molecules lead to the formation of a two-dimensional supramolecular network lying parallel to the ab plane. The sheets are stacked along the c-axis direction and linked by F and weak – stacking interactions between pairs of inversion-related A molecules, forming a three-dimensional supramolecular network. The photoluminescence quantum efﬁciency of the title compound in the blue–green region of the visible region (max = 517 and 544 nm) is estimated to be 0.2–0.3, indicating that the title compound could be a suitable candidate as the emitting material in organic light-emitting diode (OLED) applications. CCDC reference: 2053861 Supporting information: this article has supporting information at journals.iucr.org/e The C,N-chelating 2,30-bipyridine-based transition-metal compounds have attracted much interest because of their wide applications as biological labels, photosensitizers in water reduction, sensors and organic light-emitting diodes (OLEDs) (Zaen et al., 2019a,b). Especially, highly efﬁcient phospho- rescent metal complexes containing IrIII and PtII can be synthesized by using 2,30-bipyridine as ligand, which feature a high triplet-state energy (Lee et al., 2018). In terms of the efﬁciency and stability of OLEDs, tetradentate ligand-based PtII complexes are known to be very good candidates as triplet emitters (Wang & Wang, 2019). research communications research communications research communications Synthesis, crystal structure and photophysical properties of chlorido[2-(2000,6000-difluoro-2,3000-bipyri- din-6-yl-jN1)-6-(pyridin-2-yloxy-jN)phenyl-jC1]- platinum(II) ISSN 2056-9890 research communications Table 1 Selected geometric parameters (A˚ , ). Pt1—C11 1.949 (4) Pt2—C32 1.948 (4) Pt1—N1 2.035 (3) Pt2—N4 2.042 (3) Pt1—N2 2.040 (3) Pt2—N5 2.049 (3) Pt1—Cl1 2.4193 (8) Pt2—Cl2 2.4154 (8) C11—Pt1—N1 90.92 (14) C32—Pt2—N4 90.87 (14) C11—Pt1—N2 80.86 (14) C32—Pt2—N5 80.98 (13) N1—Pt1—N2 171.61 (12) N4—Pt2—N5 171.56 (11) C11—Pt1—Cl1 163.28 (11) C32—Pt2—Cl2 163.13 (11) N1—Pt1—Cl1 94.81 (9) N4—Pt2—Cl2 94.78 (9) N2—Pt1—Cl1 93.56 (9) N5—Pt2—Cl2 93.65 (8) synthesized its coordination metal complex containing PtII and determined its crystal structure: herein, we report the struc- tural and photophysical characteristics of chlorido[20,60-diﬂu- oro-6-[3-(pyridin-2-yloxy)phenyl]-2,30-bipyridine-3N,C,N0]- platinum(II). l slightly shorter than that [2.042 (3) A˚ ] of the Pt—N bonds because of back bonding between the metal and the anionic C atom of the ligand. The Cl1 and Cl2 atoms deviate from the mean plane consisting of the Pt and coordinated N/C atoms [r.m.s. deviations = 0.013 (1) (A) and 0.017 (1) A˚ (B)] with deviations of 0.700 (6) A˚ for A and 0.720 (6) A˚ for B. In each molecule, there are intramolecular C—H Cl/F interactions, contributing to the stabilization of the molecular structure (Table 2 and black dashed lines in Fig. 1). Moreover, an intramolecular Cl interaction [Cl1 Cg4 = 3.4537 (19) A˚ , Cl2 Cg8 = 3.455 (2) A˚ ; green dashed lines in Fig. 1; Cg4 and Cg8 are the centroids of the N3/C17–C21 and N6/C38–C42 rings, respectively] between the coordinated chloride ion and the pyridine ring with ﬂuorine substituents are also observed. Molecules A and B are interlinked by a C— H Cl interaction (Table 2 and yellow dashed line in Fig. 1). In the 6-phenyl-2,30-bipyridine system in both molecules, the phenylpyridine moieties are approximately coplanar with the dihedral angles between the pyridine ring and the attached phenyl rings being 10.01 (11) for A and 9.64 (11) for B. However, the terminal diﬂuoro-pyridine ring is tilted by 46.08 (9) for A and 46.96 (8) for B with respect to phenyl- pyridine ring plane. This distortion may be caused by the intramolecular Cl interaction described above. The pyri- dine ring of the pyridine-2-yloxy group is slightly tilted by 22.09 (13) for A and 19.70 (13) for B relative to the phenyl- pyridine ring plane. 2. Structural commentary 2. research communications Structural commentary The asymmetric unit of the title compound, Pt(C21H12F2N3O)Cl, contains two crystallographically inde- pendent molecules (A and B denote the Pt1- and Pt2- containing molecules, respectively), which adopt similar conformations (Fig. 1 and Table 1). The coordination sphere of the PtII atoms in both molecules is a distorted square-planar geometry, with the respective coordination sites occupied by one C and two N atoms from the 20,60-diﬂuoro-6-[3-(pyridin-2- yloxy)phenyl]-2,30-bipyridine ligand together with a chloride anion. The average length [1.949 (4) A˚ ] of the Pt—C bonds is Figure 1 The molecular structure of the title compound, showing the atom- numbering scheme and displacement ellipsoids at the 50% probability level. Black and green dashed lines represent intramolecular C—H Cl/ F and Cl interactions, respectively, and yellow dashed line represents intermolecular C—H Cl interaction. H atoms not involved in intra- and intermolecular interactions are not shown for clarity. Ki-Min Park,a Cheol Joo Moon,a Sanghyun Paekb and Youngjin Kangc The design of tetradentate ligands is focused on making appropriate coordination modes in order to form ﬁve or six- membered metallacycles. To achieve blue emission in Pt-based triplet emitters, two stra- tegies have been employed as follows: (i) incorporation of a high-triplet-energy moiety into the ligand framework; (ii) the breakage of -conjugation in the ligand to increase the energy gap (Fleetham et al., 2016; Kang et al., 2020). With these in mind, we have recently synthesized 20,60-diﬂuoro-6-[3- (pyridin-2-yloxy)phenyl]-2,30-bipyridine as a ligand with high triplet energy (Park et al., 2020). By using this ligand, we have Acta Cryst. (2021). E77, 107–110 107 https://doi.org/10.1107/S2056989021000128 Acta Cryst. (2021). E77, 107–110 Acta Cryst. (2021). E77, 107–110 3. Supramolecular features In the crystal structure, intermolecular C–H Cl/F hydrogen bonds (Table 2, yellow dashed lines in Figs. 1 and 2) between adjacent A and B molecules and between pairs of inversion- related B molecules lead to the formation of a two-dimen- sional supramolecular network lying parallel to the ab plane. In addition, this network is consolidated by halogen and weak – stacking interactions [red and black dashed lines in Fig. 2, respectively; F2 Cg5i = 3.819 (3); Cg6 Cg7ii = 4.022 (2) A˚ ; Cg5, Cg6 and Cg7 are the centroids of the N4/ C22–C26, C27–C32 and N5/C33–C37 rings, respectively; symmetry codes: (i) x + 2, y + 1, z; (ii) x + 2, y + 2, z] between pairs of inversion-related B molecules. These sheets are stacked along the c-axis direction and connected by F and weak – stacking interactions [sky-blue and green In addition, this network is consolidated by halogen and weak – stacking interactions [red and black dashed lines in Fig. 2, respectively; F2 Cg5i = 3.819 (3); Cg6 Cg7ii = 4.022 (2) A˚ ; Cg5, Cg6 and Cg7 are the centroids of the N4/ C22–C26, C27–C32 and N5/C33–C37 rings, respectively; symmetry codes: (i) x + 2, y + 1, z; (ii) x + 2, y + 2, z] between pairs of inversion-related B molecules. These sheets are stacked along the c-axis direction and connected by F and weak – stacking interactions [sky-blue and green Figure 1 g The molecular structure of the title compound, showing the atom- numbering scheme and displacement ellipsoids at the 50% probability level. Black and green dashed lines represent intramolecular C—H Cl/ F and Cl interactions, respectively, and yellow dashed line represents intermolecular C—H Cl interaction. H atoms not involved in intra- and intermolecular interactions are not shown for clarity. g The molecular structure of the title compound, showing the atom- numbering scheme and displacement ellipsoids at the 50% probability level. Black and green dashed lines represent intramolecular C—H Cl/ F and Cl interactions, respectively, and yellow dashed line represents intermolecular C—H Cl interaction. H atoms not involved in intra- and intermolecular interactions are not shown for clarity. 108 Park et al. [Pt(C21H12F2N3O)Cl] 108 Acta Cryst. (2021). E77, 107–110 research communications Table 2 Hydrogen-bond geometry (A˚ , ). 3. Supramolecular features D—H A D—H H A D A D—H A C1—H1 Cl1 0.95 2.46 3.175 (4) 132 C3—H3 F1i 0.95 2.53 3.474 (5) 172 C15—H15 F1 0.95 2.56 2.989 (5) 108 C22—H22 Cl2 0.95 2.45 3.170 (4) 132 C23—H23 Cl2ii 0.95 2.82 3.540 (5) 133 C24—H24 F3iii 0.95 2.50 3.443 (5) 172 C36—H36 F3 0.95 2.55 2.975 (5) 108 C41—H41 Cl1 0.95 2.79 3.684 (4) 158 Symmetry codes: (i) x; y þ 1; z; (ii) x þ 2; y þ 1; z; (iii) x 1; y; z. Figure 3 The three-dimensional supramolecular network formed through F and – stacking interactions (sky-blue and green dashed lines) between the two-dimensional networks stacked along the c-axis direction. H atoms not involved in the intermolecular interactions have been omitted for clarity. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. dashed lines in Fig. 3, respectively; F4 Cg1iii = 3.834 (3) A˚ ; Cg2 Cg3iv = 4.073 (2) A˚ ; Cg1, Cg2 and Cg3 are the centroids of the N1/C1–C5, C6–C11 and N2/C12–C16 rings, respectively; symmetry code: (iii) x + 2, y + 1, z + 1; (iv) x + 1, y + 1, z + 1] between pairs of inversion-related A molecules, resulting in the formation of a three-dimensional supra- molecular network. Figure 3 The three-dimensional supramolecular network formed through F and – stacking interactions (sky-blue and green dashed lines) between the two-dimensional networks stacked along the c-axis direction. H atoms not involved in the intermolecular interactions have been omitted for clarity. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. The three-dimensional supramolecular network formed through F and – stacking interactions (sky-blue and green dashed lines) between the two-dimensional networks stacked along the c-axis direction. H atoms not involved in the intermolecular interactions have been omitted for clarity. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. 4. Luminescent property was estimated to be 0.2–0.3 (Fig. 4, inset). Such efﬁciency is enough to use the title compound as the emitting material in organic light-emitting diode (OLED) applications. The bright blueish-green emission of the title compound in solution is dominated by phosphorescence as supported by an excited-state lifetime of more than 1 ms. Emission maxima appear at 517 and 544 nm at room temperature, as shown in Fig. 4. The emission observed in the title compound is attri- butable to an intra-ligand charge transfer (ILCT) transition mixed with a metal-to-ligand charge-transfer (MLCT) transi- tion based on previous reports (Wang & Wang, 2019). Contrary to our expectations, the title compound shows green emission. It may be that the chloride ion bound directly to the platinum ion causes this effect because 20,60-diﬂuoro-2,30-bi- pyridine (dfpypy)-based platinum complexes without chloride ions often exhibit blue emission at room temperature. The photoluminescence quantum efﬁciency of the title compound 5. Database survey A survey of SciFinder (SciFinder, 2020) for 6-[3-(pyridin-2- yloxy)phenyl]-2,30-bipyridine (i.e., the title ligand without specifying the ﬂuorine substituents), gave two hits. These are the reports of the crystal structures and photophysical prop- erties of the free ligands for 20,60-diﬂuoro-6-[3-(pyridin-2-yl- oxy)phenyl]-2,30-bipyridine and 20,60-dimethoxy-6-[3-(pyridin- 2-yloxy)phenyl]-2,30-bipyridine (Park et al., 2020). The survey revealed no exact matches for the reported structure of the Acta Cryst. (2021). E77, 107–110 Park et al. [Pt(C21H12F2N3O)Cl] 109 Figure 4 Emission spectrum of the title compound in solution at room temperature (Inset: Emission photo). Figure 2 The two-dimensional supramolecular network formed through inter- molecular C—H Cl/F hydrogen bonds (yellow dashed lines), F (red dashed lines) and – stacking (black dashed lines) interactions between aromatic rings of inversion-related B molecules. For clarity, H atoms not involved in the intermolecular interactions have been omitted. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. Figure 2 The two-dimensional supramolecular network formed through inter- molecular C—H Cl/F hydrogen bonds (yellow dashed lines), F (red dashed lines) and – stacking (black dashed lines) interactions between aromatic rings of inversion-related B molecules. For clarity, H atoms not involved in the intermolecular interactions have been omitted. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. Figure 4 Emission spectrum of the title compound in solution at room temperature (Inset: Emission photo) Figure 4 Emission spectrum of the title compound in solution at room temperature (Inset: Emission photo). Figure 2 g The two-dimensional supramolecular network formed through inter- molecular C—H Cl/F hydrogen bonds (yellow dashed lines), F (red dashed lines) and – stacking (black dashed lines) interactions between aromatic rings of inversion-related B molecules. For clarity, H atoms not involved in the intermolecular interactions have been omitted. Colour codes: violet = platinum, plum = chloride, green = ﬂuorine, red = oxygen, blue = nitrogen, grey = carbon and white = hydrogen. Figure 4 Emission spectrum of the title compound in solution at room temperature (Inset: Emission photo). Park et al. [Pt(C21H12F2N3O)Cl] 109 109 Acta Cryst. (2021). E77, 107–110 References title ligand: to the best of our knowledge, this is the ﬁrst crystal structure reported for a platinum complex with the title ligand. title ligand: to the best of our knowledge, this is the ﬁrst crystal structure reported for a platinum complex with the title ligand. Brandenburg, K. (2010). DIAMOND. Crystal Impact GbR, Bonn, Germany. Bruker (2014). APEX2, SAINT and SADABS. Bruker AXS Inc., Madison, Wisconsin, USA. , , Fleetham, T., Li, G. & Li, J. (2016). Adv. Mater. 29, 1601861. research communications Table 3 Experimental details. Crystal data Chemical formula [Pt(C21H12F2N3O)Cl] Mr 590.88 Crystal system, space group Triclinic, P1 Temperature (K) 173 a, b, c (A˚ ) 12.4924 (9), 12.5008 (9), 14.1163 (11) , , () 74.498 (3), 73.401 (3), 61.954 (3) V (A˚ 3) 1841.3 (2) Z 4 Radiation type Mo K (mm1) 7.80 Crystal size (mm) 0.46 0.32 0.24 Data collection Diffractometer Bruker APEXII CCD Absorption correction Multi-scan (SADABS; Bruker, 2014) Tmin, Tmax 0.233, 0.746 No. of measured, independent and observed [I > 2 (I)] reﬂections 34679, 9126, 7667 Rint 0.045 (sin /)max (A˚ 1) 0.667 Reﬁnement R[F 2 > 2 (F 2)], wR(F 2), S 0.028, 0.067, 1.03 No. of reﬂections 9126 No. of parameters 523 H-atom treatment H-atom parameters constrained max, min (e A˚ 3) 1.64, 1.21 Computer programs: APEX2 (Bruker, 2014), SAINT (Bruker, 2014), SHELXS97 (Sheldrick, 2008), SHELXL2014/7 (Sheldrick, 2015), DIAMOND (Brandenburg, 2010), SHELXTL (Sheldrick, 2008) and publCIF (Westrip, 2010). compound as a yellow solid in 40% yield. Orange–red crystals suitable for X-ray crystallography analysis were obtained from a CH2Cl2/hexane solution by slow evaporation. 1H NMR (400 MHz, CDCl3) 9.91 (dd, J = 6.0, 2.0 Hz, 1H), 8.20(m, 1H), 7.97 (t, J = 8.0 Hz, 1H), 7.90–7.84 (m, 2H), 7.50–7.42 (m, 2H), 7.30–7.22 (m, 2H), 7.06 (d, J = 7.6 Hz, 1H), 6.97-6.91 (m, 2H). 13C NMR (100 MHz, CDCl3) 206.5, 167.3, 156.4, 155.9, 151.0, 148.1, 146.6, 146.5, 140.1, 139.0, 125.7, 125.0, 121.0, 118.4, 118.3, 117.6, 115.9, 106.0, 105.9, 105.7, 105.6. Analysis calculated for C21H12ClF2N3OPt: C 42.69; H 2.05; N 7.11%; found: C 42.70, H 2.06, N 7.09%. [Pt(C21H12F2N3O)Cl] 590.88 Triclinic, P1 173 12.4924 (9), 12.5008 (9), 14.1163 (11) 74.498 (3), 73.401 (3), 61.954 (3) 1841.3 (2) 4 Mo K 7.80 0.46 0.32 0.24 Funding information Funding information Funding for this research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF- 2016R1D1A1B01012630 and 2018R1D1A3A03000716). This study has been undertaken with the support of a research grant of Kangwon National University in 2020. Computer programs: APEX2 (Bruker, 2014), SAINT (Bruker, 2014), SHELXS97 (Sheldrick, 2008), SHELXL2014/7 (Sheldrick, 2015), DIAMOND (Brandenburg, 2010), SHELXTL (Sheldrick, 2008) and publCIF (Westrip, 2010). 7. Refinement Crystal data, data collection and structure reﬁnement details are summarized in Table 3. All H atoms were positioned geometrically and reﬁned using a riding model: C—H = 0.95 A˚ with Uiso(H) = 1.2Ueq(C). 6. Synthesis and crystallization Kang, J., Zaen, R., Park, K.-M., Lee, K. H., Lee, J. Y. & Kang, Y. (2020). Cryst. Growth Des. 20, 6129–6138. All experiments were performed under a dry N2 atmosphere using standard Schlenk techniques. All solvents used in this study were freshly distilled over appropriate drying reagents prior to use. All starting materials were commercially purchased and used without further puriﬁcation. The 1H NMR spectrum was recorded on a JEOL 400 MHz spectrometer. The ligand, 20,30-diﬂuoro-6-[3-(pyridin-2-yloxy)phenyl]-2,30- bipyridine (Park et al., 2018, 2020) and starting material, PtCl2(PhCN)2, (Uchiyama et al., 1980) were synthesized according to previous reports. Lee, C., Zaen, R., Park, K.-M., Lee, K. H., Lee, J. Y. & Kang, Y. (2018). Organometallics, 37, 4639–4647. Park, K.-M., Moon, S.-H. & Kang, Y. (2018). Acta Cryst. E74, 1475– 1479. Park, K.-M., Yang, K., Moon, S.-H. & Kang, Y. (2020). Acta Cryst. C79, 381–388. SciFinder (2020). Chemical Abstracts Service: Colombus, OH, 2010; RN 58 08 2 ( d D b 31 2020) SciFinder (2020). Chemical Abstracts Service: Colombus, OH, 2010; RN 58-08-2 (accessed December 31, 2020). ( ) RN 58-08-2 (accessed December 31, 2020). Sheldrick G M (2008) Acta Cryst A64 112 122 Sheldrick, G. M. (2008). Acta Cryst. A64, 112–122. Sheldrick, G. M. (2015). Acta Cryst. C71, 3–8. Uchiyama, Y., Nakamura, Y., Miwa, T., Kawaguchi, S. & Okeya, S. (1980). Chem. Lett. 3, 337–338. The title compound was synthesized as follows: A mixture of the ligand (0.36 g, 1.0 mmol), PtCl2(PhCN)2 (0.47 g, 1.0 mmol) and xylene (10 ml) was reﬂuxed (433 K) for 48 h under an N2 ﬂow. The xylene was removed by distillation and the crude product was puriﬁed by silica gel column chroma- tography (CH2Cl2:hexane = 1:1, v/v) to give the title Wang, X. & Wang, S. (2019). Chem. Rec. 19, 1693–1709. Westrip, S. P. (2010). J. Appl. Cryst. 43, 920–925. Zaen, R., Kim, M., Park, K.-M., Lee, K. H., Lee, J. Y. & Kang, Y. (2019a). Dalton Trans. 48, 9734–9743. Zaen, R., Park, K.-M., Lee, K. H., Lee, J. Y. & Kang, Y. (2019b). Adv. Optical Mater. 7, 1901387. 110 Park et al. [Pt(C21H12F2N3O)Cl] Acta Cryst. (2021). E77, 107–110 supporting information supporting information supporting information p Geometry. All esds (except the esd in the dihedral angle between two l.s. planes) are estimated using the full covariance matrix. The cell esds are taken into account individually in the estimation of esds in distances, angles and torsion angles; correlations between esds in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell esds is used for estimating esds involving l.s. planes. supporting information Synthesis, crystal structure and photophysical properties of chlorido[2-(2′,6′-di- fluoro-2,3′-bipyridin-6-yl-κN1)-6-(pyridin-2-yloxy-κN)phenyl-κC1]platinum(II) Ki-Min Park, Cheol Joo Moon, Sanghyun Paek and Youngjin Kang Ki-Min Park, Cheol Joo Moon, Sanghyun Paek and Youngjin Kang Computing details Data collection: APEX2 (Bruker, 2014); cell refinement: SAINT (Bruker, 2014); data reduction: SAINT (Bruker, 2014); program(s) used to solve structure: SHELXS97 (Sheldrick, 2008); program(s) used to refine structure: SHELXL2014/7 (Sheldrick, 2015); molecular graphics: DIAMOND (Brandenburg, 2010); software used to prepare material for publication: SHELXTL (Sheldrick, 2008) and publCIF (Westrip, 2010). Chlorido[2-(2′,6′-difluoro-2,3′-bipyridin-6-yl-κN1)-6-(pyridin-2-yloxy-κN)phenyl-κC1]platinum(II) Crystal data [Pt(C21H12F2N3O)Cl] Mr = 590.88 Triclinic, P1 a = 12.4924 (9) Å b = 12.5008 (9) Å c = 14.1163 (11) Å α = 74.498 (3)° β = 73.401 (3)° γ = 61.954 (3)° V = 1841.3 (2) Å3 Z = 4 F(000) = 1120 Dx = 2.131 Mg m−3 Mo Kα radiation, λ = 0.71073 Å Cell parameters from 9258 reflections θ = 2.6–28.3° µ = 7.80 mm−1 T = 173 K Block, yellow 0.46 × 0.32 × 0.24 mm Data collection Bruker APEXII CCD diffractometer φ and ω scans Absorption correction: multi-scan (SADABS; Bruker, 2014) Tmin = 0.233, Tmax = 0.746 34679 measured reflections 9126 independent reflections 7667 reflections with I > 2σ(I) Rint = 0.045 θmax = 28.3°, θmin = 1.5° h = −16→16 k = −15→16 l = −18→18 Refinement Refinement on F2 Least-squares matrix: full R[F2 > 2σ(F2)] = 0.028 wR(F2) = 0.067 S = 1.03 9126 reflections 523 parameters 0 restraints Hydrogen site location: inferred from neighbouring sites H-atom parameters constrained w = 1/[σ2(Fo2) + (0.0327P)2] where P = (Fo2 + 2Fc2)/3 (Δ/σ)max = 0.001 Δρmax = 1.64 e Å−3 Δρmin = −1.21 e Å−3 Hydrogen site location: inferred from neighbouring sites H-atom parameters constrained w = 1/[σ2(Fo2) + (0.0327P)2] where P = (Fo2 + 2Fc2)/3 (Δ/σ)max = 0.001 Δρmax = 1.64 e Å−3 Δρmin = −1.21 e Å−3 sup-1 Acta Cryst. (2021). E77, 107-110 supporting information Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) x y z Uiso/Ueq Pt1 0.69646 (2) 0.61157 (2) 0.39509 (2) 0.03247 (5) Pt2 0.88563 (2) 0.80611 (2) 0.10492 (2) 0.03240 (5) Cl1 0.86494 (8) 0.44909 (8) 0.47237 (7) 0.0410 (2) Cl2 1.05014 (8) 0.63393 (8) 0.02853 (7) 0.0413 (2) F1 0.8999 (2) 0.1430 (2) 0.4219 (2) 0.0689 (7) F2 1.2132 (2) 0.1920 (3) 0.1978 (2) 0.0730 (8) F3 1.3562 (2) 0.6034 (2) 0.0774 (2) 0.0685 (7) F4 1.3006 (3) 0.2977 (2) 0.3009 (2) 0.0711 (7) O1 0.5471 (3) 0.9163 (2) 0.3614 (3) 0.0668 (9) O2 0.5792 (2) 0.9597 (3) 0.1333 (3) 0.0674 (9) N1 0.7262 (3) 0.7497 (3) 0.4159 (2) 0.0358 (7) N2 0.6446 (3) 0.4930 (3) 0.3661 (2) 0.0340 (7) N3 1.0540 (3) 0.1720 (3) 0.3099 (3) 0.0504 (9) N4 0.7472 (3) 0.7748 (3) 0.0861 (2) 0.0358 (7) N5 1.0042 (3) 0.8593 (3) 0.1331 (2) 0.0340 (7) N6 1.3250 (3) 0.4527 (3) 0.1888 (3) 0.0479 (8) C1 0.8297 (4) 0.7229 (4) 0.4491 (3) 0.0421 (9) H1 0.8809 0.6389 0.4688 0.050 C2 0.8652 (5) 0.8078 (4) 0.4562 (4) 0.0550 (11) H2 0.9387 0.7833 0.4797 0.066* C3 0.7916 (5) 0.9306 (4) 0.4283 (4) 0.0661 (13) H3 0.8138 0.9921 0.4322 0.079* C4 0.6870 (5) 0.9618 (4) 0.3953 (4) 0.0657 (13) H4 0.6358 1.0455 0.3748 0.079* C5 0.6559 (4) 0.8701 (4) 0.3920 (3) 0.0511 (11) C6 0.4800 (4) 0.8536 (4) 0.3637 (3) 0.0505 (10) C7 0.3573 (4) 0.9249 (4) 0.3571 (3) 0.0586 (12) H7 0.3244 1.0117 0.3499 0.070* C8 0.2835 (4) 0.8706 (4) 0.3610 (3) 0.0587 (12) H8 0.1985 0.9199 0.3585 0.070* C9 0.3308 (4) 0.7440 (4) 0.3685 (3) 0.0490 (10) H9 0.2791 0.7059 0.3720 0.059* C10 0.4556 (4) 0.6748 (4) 0.3706 (3) 0.0416 (9) C11 0.5324 (4) 0.7272 (3) 0.3681 (3) 0.0393 (8) C12 0.5184 (3) 0.5402 (3) 0.3718 (3) 0.0375 (8) C13 0.4631 (4) 0.4677 (4) 0.3732 (3) 0.0459 (10) H13 0.3762 0.5021 0.3785 0.055* C14 0.5330 (4) 0.3454 (4) 0.3668 (3) 0.0498 (10) H14 0.4949 0.2933 0.3719 0.060* Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) x y z Uiso/Ueq Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) l atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) Acta Cryst. (2021). E77, 107-110 sup-2 supporting information supporting information C15 0.6593 (4) 0.2998 (4) 0.3530 (3) 0.0463 (9) H15 0.7091 0.2161 0.3457 0.056 C16 0.7148 (4) 0.3742 (3) 0.3495 (3) 0.0370 (8) C17 0.8482 (3) 0.3332 (3) 0.3148 (3) 0.0365 (8) C18 0.9353 (4) 0.2176 (3) 0.3469 (3) 0.0461 (10) C19 1.0912 (4) 0.2433 (4) 0.2383 (3) 0.0492 (10) C20 1.0203 (4) 0.3602 (4) 0.1990 (3) 0.0475 (9) H20 1.0544 0.4082 0.1473 0.057* C21 0.8959 (4) 0.4040 (3) 0.2393 (3) 0.0426 (9) H21 0.8418 0.4847 0.2146 0.051* C22 0.7757 (3) 0.6672 (4) 0.0570 (3) 0.0408 (9) H22 0.8606 0.6136 0.0403 0.049* C23 0.6911 (4) 0.6315 (5) 0.0502 (4) 0.0566 (12) H23 0.7162 0.5556 0.0294 0.068* C24 0.5660 (4) 0.7100 (5) 0.0748 (4) 0.0658 (14) H24 0.5042 0.6880 0.0715 0.079* C25 0.5344 (4) 0.8173 (5) 0.1032 (4) 0.0655 (13) H25 0.4499 0.8718 0.1202 0.079* C26 0.6258 (4) 0.8477 (4) 0.1076 (3) 0.0500 (10) C27 0.6426 (4) 1.0252 (4) 0.1338 (3) 0.0480 (10) C28 0.5712 (4) 1.1484 (4) 0.1399 (3) 0.0576 (12) H28 0.4841 1.1823 0.1463 0.069* C29 0.6266 (4) 1.2206 (4) 0.1368 (3) 0.0589 (12) H29 0.5775 1.3056 0.1394 0.071* C30 0.7530 (4) 1.1732 (4) 0.1298 (3) 0.0507 (10) H30 0.7910 1.2249 0.1261 0.061* C31 0.8231 (4) 1.0476 (4) 0.1284 (3) 0.0398 (9) C32 0.7694 (3) 0.9716 (3) 0.1304 (3) 0.0380 (8) C33 0.9573 (4) 0.9852 (4) 0.1276 (3) 0.0398 (9) C34 1.0291 (4) 1.0404 (4) 0.1256 (3) 0.0439 (9) H34 0.9949 1.1272 0.1193 0.053* C35 1.1513 (4) 0.9709 (4) 0.1327 (3) 0.0499 (10) H35 1.2035 1.0087 0.1281 0.060* C36 1.1962 (4) 0.8450 (4) 0.1465 (3) 0.0466 (9) H36 1.2797 0.7955 0.1541 0.056* C37 1.1221 (3) 0.7895 (4) 0.1496 (3) 0.0370 (8) C38 1.1627 (3) 0.6569 (3) 0.1841 (3) 0.0349 (8) C39 1.2793 (3) 0.5693 (4) 0.1529 (3) 0.0452 (10) C40 1.2510 (4) 0.4174 (4) 0.2611 (3) 0.0494 (10) C41 1.1313 (4) 0.4895 (4) 0.2994 (3) 0.0463 (9) H41 1.0810 0.4570 0.3502 0.056* C42 1.0885 (3) 0.6118 (4) 0.2596 (3) 0.0414 (9) H42 1.0066 0.6665 0.2843 0.050* sup-3 Acta Cryst. (2021). supporting information E77, 107-110 Atomic displacement parameters (Å2) U11 U22 U33 U12 U13 U23 Pt1 0.03777 (9) 0.02938 (8) 0.03200 (8) −0.01344 (6) −0.01039 (6) −0.00506 (6) supporting information pp g Pt2 0.02768 (8) 0.03840 (9) 0.03250 (8) −0.01276 (6) −0.00509 (6) −0.01087 (6) Cl1 0.0438 (5) 0.0350 (4) 0.0486 (5) −0.0142 (4) −0.0198 (4) −0.0065 (4) Cl2 0.0340 (5) 0.0435 (5) 0.0498 (5) −0.0123 (4) −0.0076 (4) −0.0212 (4) F1 0.0752 (18) 0.0410 (13) 0.0822 (19) −0.0289 (13) −0.0188 (15) 0.0153 (13) F2 0.0478 (15) 0.0675 (18) 0.093 (2) −0.0155 (14) −0.0027 (14) −0.0253 (15) F3 0.0387 (13) 0.0711 (17) 0.0841 (19) −0.0261 (13) 0.0130 (12) −0.0154 (14) F4 0.0702 (18) 0.0477 (15) 0.0856 (19) −0.0189 (14) −0.0214 (15) −0.0003 (14) O1 0.071 (2) 0.0312 (15) 0.105 (3) −0.0161 (15) −0.047 (2) 0.0014 (16) O2 0.0295 (15) 0.071 (2) 0.104 (3) −0.0123 (15) −0.0051 (15) −0.0447 (19) N1 0.0453 (18) 0.0280 (15) 0.0366 (16) −0.0167 (14) −0.0065 (13) −0.0082 (12) N2 0.0411 (17) 0.0318 (16) 0.0313 (16) −0.0158 (14) −0.0099 (13) −0.0047 (12) N3 0.050 (2) 0.0360 (18) 0.067 (2) −0.0129 (17) −0.0152 (18) −0.0159 (17) N4 0.0288 (15) 0.0432 (18) 0.0364 (16) −0.0142 (14) −0.0073 (12) −0.0088 (14) N5 0.0321 (16) 0.0387 (17) 0.0330 (16) −0.0136 (14) −0.0054 (12) −0.0122 (13) N6 0.0372 (18) 0.045 (2) 0.061 (2) −0.0130 (16) −0.0155 (17) −0.0106 (17) C1 0.048 (2) 0.042 (2) 0.045 (2) −0.0228 (19) −0.0103 (18) −0.0106 (17) C2 0.064 (3) 0.051 (3) 0.062 (3) −0.030 (2) −0.014 (2) −0.015 (2) C3 0.090 (4) 0.049 (3) 0.079 (3) −0.040 (3) −0.024 (3) −0.012 (2) C4 0.084 (3) 0.031 (2) 0.092 (4) −0.022 (2) −0.039 (3) −0.006 (2) C5 0.062 (3) 0.037 (2) 0.052 (3) −0.015 (2) −0.018 (2) −0.0069 (19) C6 0.056 (3) 0.037 (2) 0.053 (3) −0.011 (2) −0.021 (2) −0.0040 (19) C7 0.063 (3) 0.041 (2) 0.066 (3) −0.006 (2) −0.031 (2) −0.008 (2) C8 0.046 (2) 0.061 (3) 0.057 (3) −0.008 (2) −0.022 (2) −0.006 (2) C9 0.041 (2) 0.056 (3) 0.047 (2) −0.011 (2) −0.0166 (18) −0.011 (2) C10 0.045 (2) 0.045 (2) 0.034 (2) −0.0149 (19) −0.0134 (17) −0.0064 (17) C11 0.044 (2) 0.038 (2) 0.0312 (19) −0.0135 (18) −0.0126 (16) −0.0006 (16) C12 0.041 (2) 0.042 (2) 0.0297 (19) −0.0163 (18) −0.0118 (16) −0.0026 (16) C13 0.045 (2) 0.061 (3) 0.041 (2) −0.025 (2) −0.0128 (18) −0.0124 (19) C14 0.063 (3) 0.054 (3) 0.052 (2) −0.038 (2) −0.018 (2) −0.007 (2) C15 0.058 (3) 0.042 (2) 0.046 (2) −0.025 (2) −0.0131 (19) −0.0092 (18) C16 0.049 (2) 0.0316 (18) 0.0343 (19) −0.0173 (17) −0.0125 (16) −0.0048 (15) C17 0.045 (2) 0.0310 (18) 0.037 (2) −0.0149 (17) −0.0091 (16) −0.0110 (16) C18 0.055 (3) 0.034 (2) 0.053 (3) −0.020 (2) −0.012 (2) −0.0076 (18) C19 0.043 (2) 0.047 (2) 0.058 (3) −0.013 (2) −0.010 (2) −0.022 (2) C20 0.052 (2) 0.052 (2) 0.041 (2) −0.026 (2) −0.0040 (18) −0.0090 (19) C21 0.054 (2) 0.036 (2) 0.035 (2) −0.0141 (18) −0.0130 (17) −0.0053 (16) C22 0.0339 (19) 0.048 (2) 0.047 (2) −0.0187 (18) −0.0086 (16) −0.0132 (18) C23 0.051 (3) 0.067 (3) 0.065 (3) −0.033 (2) −0.011 (2) −0.016 (2) C24 0.042 (3) 0.094 (4) 0.081 (3) −0.039 (3) −0.010 (2) −0.027 (3) C25 0.032 (2) 0.081 (3) 0.091 (4) −0.020 (2) −0.008 (2) −0.038 (3) C26 0.036 (2) 0.064 (3) 0.052 (3) −0.017 (2) −0.0089 (18) −0.020 (2) C27 0.036 (2) 0.055 (3) 0.048 (2) −0.011 (2) −0.0038 (18) −0.021 (2) C28 0.039 (2) 0.063 (3) 0.064 (3) −0.004 (2) −0.011 (2) −0.032 (2) C29 0.055 (3) 0.045 (2) 0.060 (3) −0.001 (2) −0.010 (2) −0.024 (2) C30 0.058 (3) 0.044 (2) 0.045 (2) −0.013 (2) −0.008 (2) −0.0184 (19) C31 0.040 (2) 0.040 (2) 0.037 (2) −0.0118 (18) −0.0059 (16) −0.0147 (17) C32 0.035 (2) 0.042 (2) 0.0330 (19) −0.0118 (17) −0.0047 (15) −0.0111 (16) C33 0.042 (2) 0.043 (2) 0.033 (2) −0.0153 (18) 0.0004 (16) −0.0188 (17) Acta Cryst. supporting information E77, 107-110 sup-4 supporting information pp g C34 0.052 (2) 0.042 (2) 0.040 (2) −0.019 (2) −0.0089 (18) −0.0125 (17) C35 0.053 (3) 0.060 (3) 0.055 (3) −0.035 (2) −0.0053 (19) −0.019 (2) C36 0.036 (2) 0.058 (3) 0.052 (2) −0.021 (2) −0.0093 (17) −0.015 (2) C37 0.036 (2) 0.047 (2) 0.0302 (19) −0.0164 (18) −0.0043 (15) −0.0132 (16) C38 0.0292 (18) 0.044 (2) 0.0349 (19) −0.0143 (16) −0.0083 (15) −0.0107 (16) C39 0.032 (2) 0.062 (3) 0.047 (2) −0.025 (2) −0.0054 (17) −0.011 (2) C40 0.050 (2) 0.045 (2) 0.058 (3) −0.017 (2) −0.022 (2) −0.008 (2) C41 0.050 (2) 0.055 (3) 0.035 (2) −0.025 (2) −0.0102 (18) −0.0031 (18) C42 0.035 (2) 0.053 (2) 0.034 (2) −0.0157 (18) −0.0054 (15) −0.0114 (17) Geometric parameters (Å, º) Pt1—C11 1.949 (4) C12—C13 1.368 (5) Pt1—N1 2.035 (3) C13—C14 1.370 (6) Pt1—N2 2.040 (3) C13—H13 0.9500 Pt1—Cl1 2.4193 (8) C14—C15 1.375 (6) Pt2—C32 1.948 (4) C14—H14 0.9500 Pt2—N4 2.042 (3) C15—C16 1.381 (5) Pt2—N5 2.049 (3) C15—H15 0.9500 Pt2—Cl2 2.4154 (8) C16—C17 1.465 (5) F1—C18 1.334 (5) C17—C21 1.380 (5) F2—C19 1.356 (5) C17—C18 1.389 (5) F3—C39 1.341 (4) C19—C20 1.363 (6) F4—C40 1.347 (5) C20—C21 1.379 (5) O1—C5 1.354 (5) C20—H20 0.9500 O1—C6 1.380 (5) C21—H21 0.9500 O2—C26 1.347 (5) C22—C23 1.360 (6) O2—C27 1.385 (5) C22—H22 0.9500 N1—C5 1.344 (5) C23—C24 1.398 (6) N1—C1 1.364 (5) C23—H23 0.9500 N2—C16 1.369 (5) C24—C25 1.348 (7) N2—C12 1.385 (5) C24—H24 0.9500 N3—C19 1.292 (6) C25—C26 1.382 (6) N3—C18 1.311 (5) C25—H25 0.9500 N4—C26 1.344 (5) C27—C28 1.382 (6) N4—C22 1.366 (5) C27—C32 1.391 (5) N5—C37 1.365 (5) C28—C29 1.357 (7) N5—C33 1.386 (5) C28—H28 0.9500 N6—C40 1.301 (6) C29—C30 1.384 (6) N6—C39 1.302 (5) C29—H29 0.9500 C1—C2 1.364 (6) C30—C31 1.395 (5) C1—H1 0.9500 C30—H30 0.9500 C2—C3 1.383 (6) C31—C32 1.389 (5) C2—H2 0.9500 C31—C33 1.477 (5) C3—C4 1.359 (7) C33—C34 1.355 (5) C3—H3 0.9500 C34—C35 1.374 (6) C4—C5 1.388 (6) C34—H34 0.9500 C4—H4 0.9500 C35—C36 1.378 (6) C34 0.052 (2) 0.042 (2) 0.040 (2) −0.019 (2) −0.0089 (18) −0.0125 (17) C35 0.053 (3) 0.060 (3) 0.055 (3) −0.035 (2) −0.0053 (19) −0.019 (2) C36 0.036 (2) 0.058 (3) 0.052 (2) −0.021 (2) −0.0093 (17) −0.015 (2) C37 0.036 (2) 0.047 (2) 0.0302 (19) −0.0164 (18) −0.0043 (15) −0.0132 (16) C38 0.0292 (18) 0.044 (2) 0.0349 (19) −0.0143 (16) −0.0083 (15) −0.0107 (16) C39 0.032 (2) 0.062 (3) 0.047 (2) −0.025 (2) −0.0054 (17) −0.011 (2) C40 0.050 (2) 0.045 (2) 0.058 (3) −0.017 (2) −0.022 (2) −0.008 (2) C41 0.050 (2) 0.055 (3) 0.035 (2) −0.025 (2) −0.0102 (18) −0.0031 (18) C42 0.035 (2) 0.053 (2) 0.034 (2) −0.0157 (18) −0.0054 (15) −0.0114 (17) Acta Cryst. supporting information (2021). supporting information (2021). E77, 107-110 sup-5 supporting information supporting information C3—C4—C5 119.3 (4) C30—C29—H29 119.2 C3—C4—H4 120.4 C29—C30—C31 118.4 (4) C5—C4—H4 120.4 C29—C30—H30 120.8 N1—C5—O1 124.2 (4) C31—C30—H30 120.8 N1—C5—C4 123.8 (4) C32—C31—C30 121.4 (4) O1—C5—C4 112.0 (4) C32—C31—C33 115.2 (3) O1—C6—C7 115.9 (4) C30—C31—C33 123.3 (4) O1—C6—C11 122.8 (4) C31—C32—C27 117.6 (4) C7—C6—C11 121.3 (4) C31—C32—Pt2 114.9 (3) C8—C7—C6 119.9 (4) C27—C32—Pt2 126.6 (3) C8—C7—H7 120.1 C34—C33—N5 122.0 (4) C6—C7—H7 120.1 C34—C33—C31 125.6 (4) C7—C8—C9 120.9 (4) N5—C33—C31 112.4 (3) C7—C8—H8 119.5 C33—C34—C35 120.1 (4) C9—C8—H8 119.5 C33—C34—H34 120.0 C10—C9—C8 118.0 (4) C35—C34—H34 120.0 C10—C9—H9 121.0 C34—C35—C36 118.3 (4) C8—C9—H9 121.0 C34—C35—H35 120.8 C11—C10—C9 122.3 (4) C36—C35—H35 120.8 C11—C10—C12 114.5 (3) C35—C36—C37 121.1 (4) C9—C10—C12 123.1 (4) C35—C36—H36 119.4 C10—C11—C6 117.4 (4) C37—C36—H36 119.4 C10—C11—Pt1 115.3 (3) N5—C37—C36 120.1 (4) C6—C11—Pt1 126.4 (3) N5—C37—C38 119.0 (3) C13—C12—N2 121.7 (4) C36—C37—C38 120.3 (3) C13—C12—C10 125.7 (4) C39—C38—C42 114.6 (3) N2—C12—C10 112.5 (3) C39—C38—C37 124.0 (3) C12—C13—C14 120.2 (4) C42—C38—C37 121.1 (3) C12—C13—H13 119.9 N6—C39—F3 114.3 (4) C14—C13—H13 119.9 N6—C39—C38 126.7 (4) C13—C14—C15 118.6 (4) F3—C39—C38 118.9 (4) C13—C14—H14 120.7 N6—C40—F4 115.0 (4) C15—C14—H14 120.7 N6—C40—C41 126.4 (4) C14—C15—C16 120.9 (4) F4—C40—C41 118.6 (4) C14—C15—H15 119.6 C40—C41—C42 115.7 (4) C16—C15—H15 119.6 C40—C41—H41 122.1 N2—C16—C15 120.5 (4) C42—C41—H41 122.1 N2—C16—C17 118.4 (3) C41—C42—C38 121.2 (4) C15—C16—C17 120.5 (3) C41—C42—H42 119.4 C21—C17—C18 114.7 (4) C38—C42—H42 119.4 C5—N1—C1—C2 −1.9 (6) C26—N4—C22—C23 −0.8 (6) Pt1—N1—C1—C2 173.2 (3) Pt2—N4—C22—C23 174.2 (3) N1—C1—C2—C3 0.3 (7) N4—C22—C23—C24 −0.1 (7) C1—C2—C3—C4 0.2 (7) C22—C23—C24—C25 0.5 (8) C2—C3—C4—C5 1.0 (8) C23—C24—C25—C26 0.0 (8) C1—N1—C5—O1 −177.3 (4) C22—N4—C26—O2 −176.9 (4) Pt1—N1—C5—O1 8.0 (6) Pt2—N4—C26—O2 8.5 (6) 119.3 (4) C30—C29—H29 119.2 120.4 C29—C30—C31 118.4 (4) 120.4 C29—C30—H30 120.8 124.2 (4) C31—C30—H30 120.8 123.8 (4) C32—C31—C30 121.4 (4) 112.0 (4) C32—C31—C33 115.2 (3) 115.9 (4) C30—C31—C33 123.3 (4) 122.8 (4) C31—C32—C27 117.6 (4) 121.3 (4) C31—C32—Pt2 114.9 (3) 119.9 (4) C27—C32—Pt2 126.6 (3) 120.1 C34—C33—N5 122.0 (4) 120.1 C34—C33—C31 125.6 (4) 120.9 (4) N5—C33—C31 112.4 (3) 119.5 C33—C34—C35 120.1 (4) 119.5 C33—C34—H34 120.0 118.0 (4) C35—C34—H34 120.0 121.0 C34—C35—C36 118.3 (4) 121.0 C34—C35—H35 120.8 122.3 (4) C36—C35—H35 120.8 114.5 (3) C35—C36—C37 121.1 (4) 123.1 (4) C35—C36—H36 119.4 117.4 (4) C37—C36—H36 119.4 115.3 (3) N5—C37—C36 120.1 (4) 126.4 (3) N5—C37—C38 119.0 (3) 121.7 (4) C36—C37—C38 120.3 (3) 125.7 (4) C39—C38—C42 114.6 (3) 112.5 (3) C39—C38—C37 124.0 (3) 120.2 (4) C42—C38—C37 121.1 (3) 119.9 N6—C39—F3 114.3 (4) 119.9 N6—C39—C38 126.7 (4) 118.6 (4) F3—C39—C38 118.9 (4) 120.7 N6—C40—F4 115.0 (4) 120.7 N6—C40—C41 126.4 (4) 120.9 (4) F4—C40—C41 118.6 (4) 119.6 C40—C41—C42 115.7 (4) 119.6 C40—C41—H41 122.1 120.5 (4) C42—C41—H41 122.1 118.4 (3) C41—C42—C38 121.2 (4) 120.5 (3) C41—C42—H42 119.4 114.7 (4) C38—C42—H42 119.4 −1.9 (6) C26—N4—C22—C23 −0.8 (6) 173.2 (3) Pt2—N4—C22—C23 174.2 (3) 0.3 (7) N4—C22—C23—C24 −0.1 (7) 0.2 (7) C22—C23—C24—C25 0.5 (8) 1.0 (8) C23—C24—C25—C26 0.0 (8) −177.3 (4) C22—N4—C26—O2 −176.9 (4) 8.0 (6) Pt2—N4—C26—O2 8.5 (6) Acta Cryst. supporting information supporting information C6—C7 1.380 (6) C35—H35 0.9500 C6—C11 1.388 (5) C36—C37 1.381 (5) C7—C8 1.362 (7) C36—H36 0.9500 C7—H7 0.9500 C37—C38 1.470 (5) C8—C9 1.391 (6) C38—C39 1.380 (5) C8—H8 0.9500 C38—C42 1.386 (5) C9—C10 1.387 (5) C40—C41 1.369 (6) C9—H9 0.9500 C41—C42 1.371 (5) C10—C11 1.383 (6) C41—H41 0.9500 C10—C12 1.482 (5) C42—H42 0.9500 C11—Pt1—N1 90.92 (14) C21—C17—C16 121.3 (3) C11—Pt1—N2 80.86 (14) C18—C17—C16 123.7 (4) N1—Pt1—N2 171.61 (12) N3—C18—F1 114.5 (4) C11—Pt1—Cl1 163.28 (11) N3—C18—C17 125.7 (4) N1—Pt1—Cl1 94.81 (9) F1—C18—C17 119.7 (4) N2—Pt1—Cl1 93.56 (9) N3—C19—F2 114.7 (4) C32—Pt2—N4 90.87 (14) N3—C19—C20 126.8 (4) C32—Pt2—N5 80.98 (13) F2—C19—C20 118.4 (4) N4—Pt2—N5 171.56 (11) C19—C20—C21 115.4 (4) C32—Pt2—Cl2 163.13 (11) C19—C20—H20 122.3 N4—Pt2—Cl2 94.78 (9) C21—C20—H20 122.3 N5—Pt2—Cl2 93.65 (8) C20—C21—C17 121.5 (4) C5—O1—C6 127.3 (3) C20—C21—H21 119.3 C26—O2—C27 128.0 (3) C17—C21—H21 119.3 C5—N1—C1 114.8 (3) C23—C22—N4 124.6 (4) C5—N1—Pt1 125.2 (3) C23—C22—H22 117.7 C1—N1—Pt1 119.8 (2) N4—C22—H22 117.7 C16—N2—C12 117.5 (3) C22—C23—C24 118.0 (4) C16—N2—Pt1 129.2 (3) C22—C23—H23 121.0 C12—N2—Pt1 113.1 (2) C24—C23—H23 121.0 C19—N3—C18 115.8 (4) C25—C24—C23 119.1 (4) C26—N4—C22 115.1 (3) C25—C24—H24 120.5 C26—N4—Pt2 125.5 (3) C23—C24—H24 120.5 C22—N4—Pt2 119.2 (2) C24—C25—C26 119.6 (4) C37—N5—C33 117.7 (3) C24—C25—H25 120.2 C37—N5—Pt2 129.0 (2) C26—C25—H25 120.2 C33—N5—Pt2 113.1 (2) N4—C26—O2 124.0 (4) C40—N6—C39 115.3 (4) N4—C26—C25 123.7 (4) C2—C1—N1 124.8 (4) O2—C26—C25 112.2 (4) C2—C1—H1 117.6 C28—C27—O2 115.6 (4) N1—C1—H1 117.6 C28—C27—C32 121.5 (4) C1—C2—C3 118.4 (4) O2—C27—C32 122.9 (4) C1—C2—H2 120.8 C29—C28—C27 119.5 (4) C3—C2—H2 120.8 C29—C28—H28 120.3 C4—C3—C2 118.9 (4) C27—C28—H28 120.3 C4—C3—H3 120.5 C28—C29—C30 121.5 (4) C2—C3—H3 120.5 C28—C29—H29 119.2 Acta Cryst. (2021). E77, 107-110 sup-6 supporting information supporting information supporting information supporting information pp g C1—N1—C5—C4 3.2 (6) C22—N4—C26—C25 1.3 (6) Pt1—N1—C5—C4 −171.6 (4) Pt2—N4—C26—C25 −173.3 (4) C6—O1—C5—N1 10.8 (7) C27—O2—C26—N4 6.4 (7) C6—O1—C5—C4 −169.6 (4) C27—O2—C26—C25 −172.0 (4) C3—C4—C5—N1 −2.9 (8) C24—C25—C26—N4 −0.9 (8) C3—C4—C5—O1 177.5 (5) C24—C25—C26—O2 177.5 (5) C5—O1—C6—C7 161.3 (4) C26—O2—C27—C28 165.0 (4) C5—O1—C6—C11 −21.3 (7) C26—O2—C27—C32 −16.3 (7) O1—C6—C7—C8 −178.6 (4) O2—C27—C28—C29 −177.4 (4) C11—C6—C7—C8 4.0 (7) C32—C27—C28—C29 3.9 (7) C6—C7—C8—C9 −2.0 (7) C27—C28—C29—C30 −1.5 (7) C7—C8—C9—C10 −0.8 (7) C28—C29—C30—C31 −1.5 (7) C8—C9—C10—C11 1.7 (6) C29—C30—C31—C32 2.1 (6) C8—C9—C10—C12 −176.0 (4) C29—C30—C31—C33 −176.0 (4) C9—C10—C11—C6 0.2 (6) C30—C31—C32—C27 0.1 (6) C12—C10—C11—C6 178.1 (3) C33—C31—C32—C27 178.4 (3) C9—C10—C11—Pt1 169.8 (3) C30—C31—C32—Pt2 169.9 (3) C12—C10—C11—Pt1 −12.4 (4) C33—C31—C32—Pt2 −11.8 (4) O1—C6—C11—C10 179.7 (4) C28—C27—C32—C31 −3.2 (6) C7—C6—C11—C10 −3.1 (6) O2—C27—C32—C31 178.2 (4) O1—C6—C11—Pt1 11.4 (6) C28—C27—C32—Pt2 −171.7 (3) C7—C6—C11—Pt1 −171.3 (3) O2—C27—C32—Pt2 9.7 (6) C16—N2—C12—C13 7.8 (5) C37—N5—C33—C34 8.4 (5) Pt1—N2—C12—C13 −167.8 (3) Pt2—N5—C33—C34 −167.7 (3) C16—N2—C12—C10 −169.1 (3) C37—N5—C33—C31 −169.4 (3) Pt1—N2—C12—C10 15.3 (4) Pt2—N5—C33—C31 14.5 (4) C11—C10—C12—C13 −179.3 (4) C32—C31—C33—C34 179.9 (4) C9—C10—C12—C13 −1.5 (6) C30—C31—C33—C34 −1.8 (6) C11—C10—C12—N2 −2.5 (5) C32—C31—C33—N5 −2.3 (5) C9—C10—C12—N2 175.3 (3) C30—C31—C33—N5 175.9 (3) N2—C12—C13—C14 −1.3 (6) N5—C33—C34—C35 −2.4 (6) C10—C12—C13—C14 175.2 (4) C31—C33—C34—C35 175.1 (4) C12—C13—C14—C15 −4.0 (6) C33—C34—C35—C36 −3.1 (6) C13—C14—C15—C16 2.6 (6) C34—C35—C36—C37 2.4 (6) C12—N2—C16—C15 −9.2 (5) C33—N5—C37—C36 −8.9 (5) Pt1—N2—C16—C15 165.6 (3) Pt2—N5—C37—C36 166.4 (3) C12—N2—C16—C17 162.7 (3) C33—N5—C37—C38 162.7 (3) Pt1—N2—C16—C17 −22.5 (5) Pt2—N5—C37—C38 −21.9 (5) C14—C15—C16—N2 4.2 (6) C35—C36—C37—N5 3.8 (6) C14—C15—C16—C17 −167.5 (4) C35—C36—C37—C38 −167.8 (4) N2—C16—C17—C21 −45.1 (5) N5—C37—C38—C39 142.1 (4) C15—C16—C17—C21 126.7 (4) C36—C37—C38—C39 −46.3 (5) N2—C16—C17—C18 141.4 (4) N5—C37—C38—C42 −44.8 (5) C15—C16—C17—C18 −46.8 (5) C36—C37—C38—C42 126.8 (4) C19—N3—C18—F1 −178.8 (4) C40—N6—C39—F3 −178.8 (3) C19—N3—C18—C17 1.0 (6) C40—N6—C39—C38 0.3 (6) C21—C17—C18—N3 −2.0 (6) C42—C38—C39—N6 −1.7 (6) C16—C17—C18—N3 171.9 (4) C37—C38—C39—N6 171.7 (4) pp g 3.2 (6) C22—N4—C26—C25 1.3 (6) −171.6 (4) Pt2—N4—C26—C25 −173.3 (4) 10.8 (7) C27—O2—C26—N4 6.4 (7) −169.6 (4) C27—O2—C26—C25 −172.0 (4) −2.9 (8) C24—C25—C26—N4 −0.9 (8) 177.5 (5) C24—C25—C26—O2 177.5 (5) 161.3 (4) C26—O2—C27—C28 165.0 (4) −21.3 (7) C26—O2—C27—C32 −16.3 (7) −178.6 (4) O2—C27—C28—C29 −177.4 (4) 4.0 (7) C32—C27—C28—C29 3.9 (7) −2.0 (7) C27—C28—C29—C30 −1.5 (7) −0.8 (7) C28—C29—C30—C31 −1.5 (7) 1.7 (6) C29—C30—C31—C32 2.1 (6) −176.0 (4) C29—C30—C31—C33 −176.0 (4) 0.2 (6) C30—C31—C32—C27 0.1 (6) 178.1 (3) C33—C31—C32—C27 178.4 (3) 169.8 (3) C30—C31—C32—Pt2 169.9 (3) −12.4 (4) C33—C31—C32—Pt2 −11.8 (4) 179.7 (4) C28—C27—C32—C31 −3.2 (6) −3.1 (6) O2—C27—C32—C31 178.2 (4) 11.4 (6) C28—C27—C32—Pt2 −171.7 (3) −171.3 (3) O2—C27—C32—Pt2 9.7 (6) 7.8 (5) C37—N5—C33—C34 8.4 (5) −167.8 (3) Pt2—N5—C33—C34 −167.7 (3) −169.1 (3) C37—N5—C33—C31 −169.4 (3) 15.3 (4) Pt2—N5—C33—C31 14.5 (4) −179.3 (4) C32—C31—C33—C34 179.9 (4) −1.5 (6) C30—C31—C33—C34 −1.8 (6) −2.5 (5) C32—C31—C33—N5 −2.3 (5) 175.3 (3) C30—C31—C33—N5 175.9 (3) −1.3 (6) N5—C33—C34—C35 −2.4 (6) 175.2 (4) C31—C33—C34—C35 175.1 (4) −4.0 (6) C33—C34—C35—C36 −3.1 (6) 2.6 (6) C34—C35—C36—C37 2.4 (6) −9.2 (5) C33—N5—C37—C36 −8.9 (5) 165.6 (3) Pt2—N5—C37—C36 166.4 (3) 162.7 (3) C33—N5—C37—C38 162.7 (3) −22.5 (5) Pt2—N5—C37—C38 −21.9 (5) 4.2 (6) C35—C36—C37—N5 3.8 (6) −167.5 (4) C35—C36—C37—C38 −167.8 (4) −45.1 (5) N5—C37—C38—C39 142.1 (4) 126.7 (4) C36—C37—C38—C39 −46.3 (5) 141.4 (4) N5—C37—C38—C42 −44.8 (5) −46.8 (5) C36—C37—C38—C42 126.8 (4) −178.8 (4) C40—N6—C39—F3 −178.8 (3) 1.0 (6) C40—N6—C39—C38 0.3 (6) −2.0 (6) C42—C38—C39—N6 −1.7 (6) 171.9 (4) C37—C38—C39—N6 171.7 (4) Acta Cryst. supporting information (2021). E77, 107-110 sup-7 Symmetry codes: (i) x, y+1, z; (ii) −x+2, −y+1, −z; (iii) x−1, y, z. supporting information (2021). E77, 107-110 sup-8 C21—C17—C18—F1 177.7 (3) C42—C38—C39—F3 177.4 (3) C16—C17—C18—F1 −8.4 (6) C37—C38—C39—F3 −9.2 (5) C18—N3—C19—F2 −176.8 (3) C39—N6—C40—F4 −177.1 (3) C18—N3—C19—C20 1.0 (6) C39—N6—C40—C41 2.3 (6) N3—C19—C20—C21 −1.7 (6) N6—C40—C41—C42 −3.1 (6) F2—C19—C20—C21 176.1 (3) F4—C40—C41—C42 176.4 (3) C19—C20—C21—C17 0.4 (5) C40—C41—C42—C38 1.3 (5) C18—C17—C21—C20 1.2 (5) C39—C38—C42—C41 0.8 (5) C16—C17—C21—C20 −172.8 (3) C37—C38—C42—C41 −172.9 (3) Hydrogen-bond geometry (Å, º) Hydrogen-bond geometry (Å, º) D—H···A D—H H···A D···A D—H···A C1—H1···Cl1 0.95 2.46 3.175 (4) 132 C3—H3···F1i 0.95 2.53 3.474 (5) 172 C15—H15···F1 0.95 2.56 2.989 (5) 108 C22—H22···Cl2 0.95 2.45 3.170 (4) 132 C23—H23···Cl2ii 0.95 2.82 3.540 (5) 133 C24—H24···F3iii 0.95 2.50 3.443 (5) 172 C36—H36···F3 0.95 2.55 2.975 (5) 108 C41—H41···Cl1 0.95 2.79 3.684 (4) 158 Symmetry codes: (i) x, y+1, z; (ii) −x+2, −y+1, −z; (iii) x−1, y, z. Acta Cryst. (2021). E77, 107-110 sup-9 sup-9	12,815
tel-04274811-LAURENT_Emmanuelle_vavd.txt_20	French-Science-Pile	Open Science	Various open science	2,023	Parenté, rites et territoire d’un village naxi du sud-ouest de la Chine. Sociologie. Institut National des Langues et Civilisations Orientales- INALCO PARIS - LANGUES O', 2022. Français. ⟨NNT : 2022INAL0015⟩. ⟨tel-04274811⟩	None	French	Spoken	6,727	10,148	Le rôle du charpentier débute plusieurs semaines – si ce n'est des mois – avant le dressage de la charpente. Son travail sera rémunéré plusieurs dizaines de milliers de yuan. La construction d'une nouvelle bâtisse dans la maison coûte environ 50 ou 60 000 yuan, 40 000 yuan si les hommes de la famille font eux-mêmes une partie des travaux. Une partie est le salaire versé au charpentier, le reste sert à l'achat des matériaux et à la main d’œuvre supplémentaire. Assisté des hommes de la maisonnée qui l'emploie, le charpentier procède à la sélection des arbres dans la montagne puis supervise leur coupe. En compensation du bois coupé, le maître de la maisonnée accomplit un petit culte simple aux dieux de la montagne en déposant quelques offrandes et des bâtons d’encens en remerciement. Il peut aussi tuer un coq et offrir un peu de son sang en offrande. Mis à part ce processus ritualisé de la coupe des arbres, celle-ci est plus concrètement supervisée pour l’ensemble de la population du village administratif de Wumu et de ses forêts par un responsable au sein du Comité villageois. Il est chargé de veiller au respect des forêts, à ce que les villageois prélèvent du bois de manière raisonnable et en certains lieux seulement et il surveille d’occasionnelles intrusions et vols de bois par les habitants du canton voisin. Après la coupe du bois dans la montagne et son rapatriement au village, le charpentier conçoit et fabrique la charpente dans la cour de la maison, avec l'assistance de la famille de son employeur. Tout au long de son travail, comme il est l'usage envers quiconque effectue un travail ou toute autre tâche d'entraide, cette dernière invite le charpentier aux repas matin, midi et soir. Enfin, une fois la charpente bâtie, c'est le charpentier qui supervise son dressage tout au long de la journée. Il y a au village de Wumu plusieurs charpentiers, des paysans spécialisés dans cette tâche. Les villageois font en général appel à l'un ou l'autre en fonction de leurs liens de parentés les uns avec les autres, par affinité sociale, ou selon leurs tarifs respectifs. Le charpentier et le maître de maison répartissent les rôles parmi les hommes venus aider au dressage de la charpente – parents ou amis de la famille hôte du jour –. Les profils ne s'accordent pas à toutes les tâches, cela va de l'adolescent filiforme, pour lequel c'est la première participation à un dressage de charpente, au quinquagénaire rôdé à l'exercice, en passant par les parents citadins de passage et les hommes du village dans la force de l'âge. Il y a chaque année entre cinq et dix nouveaux dressages de charpentes au village, les villageois permanents connaissent donc parfaitement la marche à suivre puisqu'ils prennent part à chacune de ces journées au nom du principe d'entraide et de réciprocité qui régit la vie sociale de Wumu. 353 Le charpentier supervise depuis le sol sans grimper sur la structure, s'assurant que les bonnes pièces trouvent chacune leurs places respectives et, à l'aide de son pendule, que les piliers porteurs soient bien verticaux. Le maître de maison reste lui aussi en bas pour remplir son rôle d'hôte et garder le recul sur l'ensemble de la situation. Sur la trentaine d'hommes participant au travail de force, cinq ou six ont pour rôle de grimper au sommet de la charpente et de superviser l'emboîtement des différentes pièces de la structure pour stabiliser l'ensemble (Photographie 26). Ces hommes se portent volontaires, ce sont souvent des jeunes non mariés (xiao huozi 小伙子) ou tout du moins des hommes fins et agiles, n'ayant pas froid aux yeux ni le vertige. Ce sont souvent des résidents permanents du village, familiers de l'exercice et rompus au dur labeur agricole. Ils se déplacent avec assurance et agilité sur les poutres surplombant la charpente, avec une désinvolture de voltigeurs, tandis que les hommes restés au sol leur font parvenir masses, planches et poutres pour immobiliser les différents plans de la structure. Au sol, les quelques dizaines d'hommes restants ont la charge de stabiliser les poutres au fur et à mesure que les plans porteurs de la charpente sont dressés. Ils assurent l'ensemble par un système de cordes jusqu'à ce que de longues hampes en bois soient fixées tout autour à tout support fixe disponible, d'un arbre voisin à un rocher, en passant par les ancrages au sol ou le pilier d'une maison voisine. Ainsi immobilisée une section de la charpente, les hommes peuvent hisser la suivante, laquelle est couchée à proximité, qui tirant, qui poussant. En tant qu'observatrice extérieure, j'ai toujours été frappée par la discipline sans faille régissant l'organisation de grands événements collectifs basés sur l'entraide villageoise. Les rôles individuels sont clairement définis, hiérarchisés et mis au service de l'action de groupe, sur la base d'une confiance mutuelle et sur les compétences respectives de chacun des participants. Les anciens sont également invités le jour du dressage de la charpente, s'ils ne participent pas au travail de force avec les jeunes, ils observent de loin toute la scène, assis à l'ombre en fumant et commentant le travail. Leur participation tient de fait davantage de la somme de leurs expériences de vie respectives, de leurs savoirs et des conseils prodigués au moment de préparer le rituel. Il y a là une coordination parfaite, qui semble s’opérer de manière naturelle, sans accros ni dispute, grâce au respect de la hiérarchie des compétences de chacun. Les dresseurs de charpente forment, le temps de cette entreprise, un tout. Parmi les autres personnes présentes le jour du dressage de la charpente, les femmes et quelques hommes reconnus pour leur compétence culinaire sont chargés de préparer les deux petits banquets auxquels se restaureront le midi tous les villageois venus aider à la journée de 354 travail, et auxquels se joindront, le soir, les membres de leurs familles, les doyens, parents et amis indisponible en journée. Le banquet est organisé comme d'ordinaire ; quatre ou cinq personnes, le plus souvent des femmes, s'activent devant de grands woks posés sur des fourneaux et font mijoter bouillons et ragoûts au-dessus du feu du foyer. Dès le matin, le petitdéjeuner avalé, les femmes de la maison s'attaquent immédiatement à l'épluchage, au tri, au lavage et à la coupe des légumes. Les ingrédients sont soigneusement préparés, la viande est découpée et marinée, un ou deux coqs sont tués pour l'occasion. Si le mobilier de la maison est insuffisant pour accueillir tous les invités, des jeunes sont envoyés chez le voisin pour emprunter une table et des tabourets. Le service et la vaisselle sont confiés aux jeunes et supervisés par les membres de la maison. En milieu de matinée, tous les travailleurs sont invités à faire une pause et à boire du thé et manger un en-cas. Les femmes de la maison ont fait frire des xi-mul-dduq83 et des baba84 sur le feu, ou des mantou85 à la vapeur. Le tout est de remplir les estomacs des travailleurs jusqu'au déjeuner qui aura lieu en début d'après-midi. Lorsque la nourriture et les plats sont prêts et servis dans des bols, il faut les présenter avec une génuflexion face à la tablette des ancêtres pour honorer ces derniers. Puis des plateaux sont chargés des bols d'offrandes et portés dans les maisons respectives des familles de la même branche lignagère ailleurs dans le village. Ce n'est qu'une fois ces offrandes retournées dans la cuisine de la maison qui organise la journée que le banquet peut commencer. Lors des jours de cérémonies, de fêtes et de banquets, comme c'est le cas notamment pour le dressage d'une charpente, il faut aussi éloigner et apaiser les esprits susceptibles de passer par là. Avant de servir le banquet, un membre de la famille dépose dans une bassine d'eau un peu de chaque plat, va en jeter le contenu à l'extérieur de la porte d'entrée de la maison et plante des bâtons d'encens à l'emplacement de l'offrande pour tenir les esprits éloignés. Pendant le banquet, le maître de maison s'assure que chacun est servi en boisson et que les hommes ont un verre d'alcool. Les personnes en cuisine mangent après les invités et les 83 Les xi-mul-dduq sont des petites galettes découpées dans un pain de pâte de riz cuit, elles sont cuites dans les cendres du feu ou frites dans un wok, saupoudrées de sel et mangées par les Naxi au petit-déjeuner ou en en-cas. En chinois, erkuai 饵块. Il s’agit par ailleurs d’une spécialité du Yunnan. 84 Les baba sont des galettes de pain naxi à base de farine de blé et de levure boulangère, généralement frites dans un wok sur le feu. C'est le petit-déjeuner de base au village. (voir Chapitre 2) 85 Les mantou 馒头 sont des petits pains à base de farine de blé cuits à la vapeur, très répandus en Chine et très appréciés aux petits-déjeuners. 355 hommes qui ont accompli le travail physique. La famille-hôte mange quant à elle en tout dernier, à la dernière tablée, une fois que tout le monde a fini de se restaurer. 2. Le processus socio-rituel du dressage de la charpente La construction d'une nouvelle maison, d'un nouveau bâtiment dans la maison ou encore d'une nouvelle porte d'entrée donne lieu à l'organisation d'une journée de travail collectif et d'entraide pour dresser une charpente. Le processus est similaire dans chacun des cas, précédé la veille par une cérémonie et conclu le jour-même par une autre cérémonie. La charpente de la maison et celle de la porte de l'enceinte de la maison peuvent être dressées le même jour ou bien à des dates différentes. Une cérémonie est nécessaire pour le hissage de la poutre faîtière symbolique tout en haut de la charpente. Lorsque la porte est fabriquée individuellement du reste de la maison, une cérémonie est donc effectuée au terme du dressage de sa charpente. Au terme du travail de force pour dresser la structure de bois, le hissage d’une poutre faîtière symbolique par-dessus la panne faîtière effective au sommet s'accompagne d'une cérémonie. La poutre symbolique est l’accessoire principal de la cérémonie qui clôt le dressage de charpente, c’est une poutre faîtière au sens rituel, social, elle ne joue en effet pas de rôle physique dans le maintien de la charpente. L’ensemble de la structure physique est tenu par la poutre faîtière physique tandis que la panne symbolique maintient l’ensemble social et cosmique. Le hissage ritualisé de cette poutre est accompli en l'honneur de l’ancêtre fondateur des charpentiers, Lu Ban qui, selon les légendes locales, serait descendu du Ciel. Cela permet également accueillir diverses divinités dongba (divinités du bois, de la vie, de la famille) ainsi que les ancêtres au sein de la nouvelle maison. Le rite apaise les mauvais esprits qui sont chassés par une longue chaîne de pétards allumée au moment du hissage de la poutre. Dans l'après-midi, une fois la charpente dressée, tandis que les hommes venus prêter main forte se reposent à l'ombre, le maître de maison, assisté et guidé par le charpentier et quelques aînés et doyens de sa famille, prépare les accessoires ainsi que les objets nécessaires au bon déroulement de la cérémonie qui clôturera la naissance de la nouvelle charpente. Ces objets doivent être sélectionnés et préparés avec minutie, car ils assureront un avenir favorable à la nouvelle maison. Un homme qui n’est ni le maître de maison, ni le Dongba, et appartenant 356 au groupe de parenté de la maisonnée, dirige la préparation de ces objets. Il charge les membres de la maisonnée de rassembler ce qu'il lui faut (Photographie 22) : un ensemble de graines (courge, fèves, blé, soja, pignons de pin), des copeaux de bois d'encens, des brisures d'or et d'autres d'argent ou bien des objets évoquant de l'or (morceaux de feuilles dorées d'emballage des paquets de cigarette) et de l'argent (une pièce de monnaie) ainsi qu'un morceau de tissu rouge. Le carré de tissu, destiné à être enveloppé autour de la poutre faîtière en son centre, doit porter le symbole du yin et du yang, les huit trigrammes ainsi que les points cardinaux. Certaines familles préfèrent acheter directement un carré de tissu déjà imprimé, d'autres invitent un homme éduqué à tracer lui-même l'ensemble des éléments sur un tissu rouge vierge avec de l'encre noire (Photographie 21). Le maître de maison fournit à l'homme ce qu'il demande. Il choisit également cinq longs fils de cinq couleurs différentes. Le regroupement des objets nécessaires se déroule sous yeux des doyens présents qui, de temps à autre, guident les plus jeunes de quelques conseils et remarques. L'autel est placé sur une table au milieu de la nouvelle charpente, sous l'emplacement de la poutre faîtière. Photographie 21 - Création du carré de tissu rouge orné des Huit Trigrammes et du Taijitu à l'encre noire en préparation de la cérémonie de hissage de la poutre faîtière symbolique (Wumu, 2018 - Photo ©Emmanuelle Laurent) Photographie 22 - Sélection des objets à insérer dans l'encoche de la panne faîtière symbolique (Wumu, 2018 - Photo ©Emmanuelle Laurent) Dans l'espoir de futures bonnes récoltes, les graines sont glissées dans une encoche taillée par le charpentier au milieu de la poutre faîtière. L'or et l'argent symbolisent ce qu'il y a de plus précieux, à placer au-dessus de la tête des habitants de la maison. L'ensemble des éléments disposés dans l'encoche de la poutre symbolise ce que l'on souhaite aux résidents de la nouvelle maison : de bonnes récoltes et de la richesse notamment. Après avoir refermé l'encoche avec le bout de bois manquant, le tissu rouge sur lequel sont tracés les Huit trigrammes (bagua 八卦) et le taijitu, le symbole du Yin et du Yang, est enroulé par-dessus. Les bagua sont un emprunt aux pratiques taoïstes. Les fils de couleur, passés à travers une pièce 357 trouée, sont enroulés et noués autour de la poutre et du carré de tissu. Pour maintenir le tout, la pièce est enfoncée à coup de maillet dans la rainure de l'encoche. La chaîne de pétards est suspendue au centre de la poutre faîtière, elle sera allumée au moment du hissage de la poutre. Le lien qui attache le pétard à la poutre est rouge, mais il peut aussi être noir, blanc, rouge, jaune, bleu ou vert. Tandis que les préparatifs suivent leur cours, deux ou trois jeunes hommes agiles se hissent encore une fois dans les hauteurs de la charpente, ils sont chargés de coller des duilian, ces sentences de bonne fortune écrites sur de longues bandes de papier rouge, sur chacune des poutres centrales de la charpente. Photographie 23 - Fixation de la poutre faîtière symbolique au sommet de la charpente (Wumu, 2018 - Photo ©Emmanuelle Laurent) La cérémonie est rapide, parfois expédiée en quelques minutes. La poutre faîtière est posée sur des supports devant la table d'offrandes installée au centre de la charpente. Le pot de cendres, les deux vases et la lampe à huile sont disposés sur cette table comme pour la cérémonie de la veille à l'instar de leur place habituelle autour de la tablette des ancêtres. Sur un côté de la table sont disposés deux ou trois outils du charpentier. Des offrandes d'alcool, de thé et de nourriture sont apportés par les femmes de la maison depuis la cuisine, l'homme qui dirige la cérémonie et guide le maître de la maison et les autres membres de sa famille verse un peu de thé et d'alcool sur la poutre puis y plante deux bâtons d'encens. Le maître de la maison et son épouse, accompagnés du maître charpentier, tournés vers la cuisine de la maison et la tablette des ancêtres, s'inclinent ou se prosternent trois fois devant la table d'offrandes avant de déposer leurs bâtons d'encens dans les vases de l'autel. Deux hommes grimpent jusqu'en haut de la charpente. La mèche de la chaîne de pétards est allumée et la poutre faîtière est hissée avec des cordes par les deux « voltigeurs » tandis que les détonations assourdissantes des 358 pétards résonnent dans l'espace alentour et effraient et chassent les esprits malveillants (Photographie 24). La poutre faîtière est attachée (Photographie 23) avec du fil de fer au-dessus de la plus haute des poutres emboîtées dans la pente. La cérémonie est destinée à inviter et accueillir la divinité de la vie, pour la prier de venir dans la maison et pour l'y accueillir. La chaîne de pétards sert à chasser les esprits malveillants. La cérémonie se conclut ainsi, le dressage de la charpente est achevé. La maison sera progressivement améliorée au fil des mois qui suivent, la famille invitera de nouveau quelques hommes pour aider à poser les tuiles, et fabriquera les briques de torchis pour construire les murs. La construction d'une maison peut s'étendre sur plusieurs années si la famille n'a pas suffisamment d'argent pour tout faire en une fois. B. Le « transfert de la maison » (jjiq-bber) et l’importation du foyer Lorsqu'une famille construit une toute nouvelle maison, son emménagement n'est pas toujours immédiatement consécutif de la fin des travaux. Après une séparation du foyer des fils mariés d'une même famille, c'est en général le fils aîné qui quitte la maison parentale avec son épouse et ses enfants pour emménager dans sa propre maison ailleurs dans le village. Le processus de séparation est longuement préparé et n'est concrétisé qu'une fois la nouvelle maison construite. Ce n'est parfois que des mois – voire des années – plus tard que la division de l’« os » est réalisée, que le partage de biens est conclu et que les membres de la famille se répartissent entre les deux foyers selon la situation. L'importation du feu depuis le foyer parental jusqu'au nouveau foyer marque la naissance de la nouvelle maison, d'un nouveau yagoq. Plusieurs expressions naxi font référence au processus de séparation, chacune désignant l'un des stades de la séparation de la famille. L'expression jjiq-bbiu86, « séparation de la maison (habitation) » désigne une séparation de la famille qui n'est pas immédiatement suivie par un déménagement dans une nouvelle maison. Elle entraîne seulement, dans un premier temps, la naissance d’une seconde maison. Entre 2016 et 2018, les deux démarches de séparation qui ont été portées à mon attention se sont conclues par un déménagement un an et demi après le dressage de la nouvelle charpente et des travaux. La phase de déménagement dans une nouvelle éralement, « maison, habitation » et « division, partage ». maison suite à une séparation de la famille est appelée jjiq-bber87 : « transfert de la maison, l'habitation ». La dernière phase, o-bbiu88, « séparation de l’os », renvoie au partage de l'os – donc à la division de la chaîne de descendance en deux lignées – et au partage entre les fils des biens, ustensiles domestiques, du bétail, des terres etc. Le fils qui demeure dans la maison parentale porte alors la responsabilité de la poursuite de la lignée principale, il vit en général avec ses deux parents, tandis que le fils qui quitte la maison devient le premier homme d'une nouvelle lignée collatérale. Le partage des biens doit être équitable, il est supervisé par deux hommes et suivi attentivement et commenté par les doyens du lignage. Lorsqu'une famille emménage dans sa nouvelle maison, elle doit apporter avec elle un certain nombre d'éléments essentiels, en plus des cendres de son ancien foyer. Au moins deux animaux doivent les suivre dans la maison : un coq pour les premières offrandes aux ancêtres ainsi qu'un chaton pour manger les souris. Il faut apporter de l'eau, du thé, de l'alcool et de quoi manger. Le premier feu de la nouvelle maison doit être allumé à partir des braises ou des flammes provenant de l'ancien foyer de la famille. Ainsi, le nouveau foyer est systématiquement construit à partir du foyer de la « maisonmère » qui est l’origine de la division de l’« os ». Cela montre que toute nouvelle maison ne peut être construite à partir de rien. La constatation de l’importation de cendres de l’ancien foyer pour fonder le nouveau n’est pas sans rappeler les écrits du sinologue néerlandais Kristofer Schipper (1934-2021) sur le principe du « fen-hsiang » (en pinyin chinois fenxiang 分香) – la « division de l’encens » (1990). Ce qu’il décrit comme une « institution largement adoptée en Chine » est utilisée « pour l’affiliation des groupes de culte et la fondation de temples » (Schipper, 1990 : 397). Cette pratique permet de créer un nouveau groupe de culte à partir d’un groupe similaire déjà existant. En effet, « lorsqu'un nouveau groupe de culte est formé, ses membres remplissent un encensoir avec les cendres recueillies dans l’[encensoir] d'un groupe de culte déjà existant du même saint. » (op. cit.). Kristofer Schipper décrit l’apport de ces cendres dans l’encensoir du nouveau groupe de culte comme le « rite fondateur » de ce celui-ci. De nos jours, nous dit le sinologue, le transfert des cendres implique « une reconnaissance et une assistance mutuelles des deux parties » (op. cit.). Le transfert de cendres est, au village, une pratique liée spécifiquement au culte des ancêtres honorés par les membres d’une maisonnée et à la tablette de la cuisine. La discussion sera approfondie sur ce sujet dans 87 88 Littéralement, « maison, habitation » et « déménagement, transfert ». Littéralement, « os » et « division, partage ». 360 le chapitre suivant portant sur le culte des ancêtres et le lien existant ces derniers et le foyer. Rappelons-le, tout événement social ou rituel donnant lieu à la performance d’un rite important, à l’organisation d’un banquet dans la maison donne lieu à un don d’offrandes aux ancêtres de la maisonnée précédé par un renouvellement des cendres dans le pot posé devant la tablette. Comme cela a été décrit à plusieurs reprises dans l’ethnographie de rites depuis le début de cette thèse, des cendres chaudes sont en effet retirées du feu du foyer de la cuisine et versées dans le pot devant la tablette, puis des braises de la même provenance sont déposées par-dessus, sur quelques allumettes taillées dans du pin. De même que le transfert de cendres des foyers d’une maison à une autre invoque une continuité des liens entre la « maison-mère » et la nouvelle maison, une filiation, le transfert de cendres entre le foyer de la maison et le foyer symbolique des ancêtres montre une volonté de renouvellement et de maintien du lien entre les membres d’une maisonnée et leurs ancêtres. Lorsqu’une famille emménage dans sa nouvelle maison après la division des frères et de la lignée principale, il n’y a, dans un premier temps, aucune tablette dans la nouvelle cuisine. Les villageois justifient cela par le fait qu’aucun membre de la maisonnée n’est encore décédé. Ce n’est qu’au premier décès qui arrivera dans la nouvelle maison qu’une tablette sera fabriquée et placée dans la cuisine au moment des funérailles. D’ici là, tous les rituels du thé matinal, les offrandes pour honorer les ancêtres epvq-esse et el-cif-loq-el-cif-nail, bien qu’effectués dans la nouvelle cuisine, seront orientés en direction de la tablette de l’ancienne maison. Ainsi, la famille d’A-Jun, dont le père a séparé l’« os » et a construit sa maison en bas du village, honore les ancêtres paternels au quotidien en se tournant vers le nord, en direction de la maison de son oncle paternel où se trouve la vieille tablette de leur lignée. Les membres de la nouvelle lignée continuent donc à rendre un culte aux ancêtres de l’« os » dont ils sont issus. La naissance d’une maison ne donne pas lieu à un commencement - d’une famille, d’un culte d’ancêtres etc. – mais à une continuation d’une descendance, d’un « groupe de lignées » ou d’un « groupe domestique ». La dynamique de la maison est liée au foyer et à la tablette des ancêtres, deux éléments constitutifs de chaque maison du village. Conclusion Yagoq a fait l’objet du présent chapitre à la lumière de la définition, des délimitations et de la dynamique de cette unité domestique au sein du village de Wumu. Il s’agit d’une unité sociale mobilisable par un groupe de yagoq-xi par l’intermédiaire du feu. Yagoq peut ainsi être exporté hors des murs du bâtiment physique provisoirement – pour une journée de culte des ancêtres dans la montagne par exemple. Dans les murs en revanche, la maison est configurée similairement d’une famille à une autre : le cœur des interactions est dans la cuisine qui d’ailleurs est, comme je le nomme, le « yagoq au sein du yagoq ». Les infrastructures sanitaires sont placées derrière la maison, tandis que les pièces essentielles – la cuisine, l’étable, le grenier, le garde-manger et les chambres – sont organisées autour de la cour carrée. Cette dernière est dotée d’un rôle fonctionnel essentiellement agricole : c’est l’endroit où sont nourris les animaux de la maisonnée, où les épis de maïs, les fèves ou les ballots de paille peuvent être mis à sécher. C’est encore l’espace où les grands banquets et les cérémonies rituelles importantes se déroulent en présence d’un grand nombre d’invités. Les maisons sont des nœuds d’interactions sociales dans le réseau formé par la communauté villageoise. Au-delà du concept de yagoq, le chapitre a cherché à décrire la maisonnée, de sa naissance à l’occupation de son espace en passant par son fonctionnement et l’organisation de la vie domestique. La dynamique de yagoq est étroitement liée aux relations de germanité, aux séparations de l’« os » qui interviennent lorsqu’une relation de consanguinité (filiation ou germanité) se dégrade au sein de la maisonnée. Nous l’avons vu, de telles circonstances entraînent d’abord la séparation de l’habitation (jjiq-bbiu), puis son transfert (jjiq-bber) avant que le groupe de filiation dit l’« os » ne soit à son tour scindé en deux (o-bbiu). La question de la séparation des frères, de l’os et de leurs maisons soulève beaucoup d’autres interrogations notamment concernant les tablettes et sur ce que la séparation implique dans le culte des ancêtres. Après division, deux maisons restent liées par le culte des ancêtres de la maison origine de la division. Les membres des deux maisonnées se rassemblent dans cette maison en plusieurs occasions dans l’année, pour honorer ensemble leurs ancêtres de lignée. Parmi le lignage Lei bbu, un groupe de maisonnées toutes issues d’un même ancêtre commun et de divisions successives de l’« os » sont restées très proches et se rassemblent à chaque repas du Nouvel an chez les uns ou les autres, à tour de rôle, ce qui leur permet ainsi de maintenir le lien entre eux et d’honorer les ancêtres respectifs de chaque maisonnée de leur groupe. Pour tenter d’y voir plus clair, le septième et dernier chapitre de cette thèse abordera les différents aspects du culte des ancêtres epvq-esse et el-cif-loq-el-cif-nail par les familles du village et discutera du devenir et des rôles des tablettes et du feu du foyer pour honorer les ancêtres de la maisonnée. Les funérailles marquant le point de départ de l’« ancestralisation » d’une personne défunte, le Chapitre 7 décrira également ce processus de naissance et de formation des nouveaux ancêtres tout en mettant en lumière les degrés de relations de parenté intervenant dans l’organisation du deuil et notamment dans l’ordre des processions qui guident et accueillent des parents endeuillés jusqu’au cercueil au début des funérailles. Chapitre 7 Ancêtres patrilinéaires : cultes et devenirs Introduction Les epvq-esse sont, je le rappelle, les ancêtres paternels masculins et féminins antérieurs de plus de quatre générations, honorés par les familles au sein des maisons. L’association des expressions epvq, « ancêtre paternel masculin », et esse, « ancêtre paternel féminin », renvoie à l’idée d’un couple ancestral, elle désigne l’ensemble des ancêtres paternels du groupe de filiation auxquels une famille de descendants rend un culte. Les el-cif-loq et el-cif-nail sont les ancêtres paternels masculins et féminins antérieurs de trois générations : les arrière-grandsparents paternels d’Ego. El-cif-loq, « arrière-grand-père paternel », et el-cif-nail, « arrièregrand-mère paternelle », sont ainsi généralement désignés comme un tout, un couple ancestral, par les descendants qui les honorent. Les epvq-essse et el-cif-loq-el-cif-nail sont représentés par des tablettes en bois, les paqweil, situées dans les cuisines. Les honorer est aussi important dans les pratiques rituelles du village qu'il est fondamental de rendre un culte aux ancêtres agnatiques dans la société rurale han. Toutes les célébrations et événements sans exception sont complétées, en parallèle, par la présentation d offrandes pour honorer les ancêtres paternels de la maisonnée. Ce septième et dernier chapitre permet de revenir sur le sujet des ancêtres patrilinéaires et des modalités de culte par lesquelles les villageois les honorent au sein de leurs maisonnées. Il en a été question tout au long de cette thèse, que ce soit dans les rituels familiaux quotidiens (Chapitre 2), par l’approche terminologique de la parenté (Chapitre 3), à travers les règles des lignages (Chapitre 5) ou encore dans le cadre de l’étude des termes vernaculaires désignant la maison et la famille (Chapitre 6). La place des ancêtres el-cif-loq-el-cif-nail et des ancêtres epvq-essse est centrale dans l’organisation sociale de la maisonnée et des lignées. Nous verrons ici les logiques relationnelles existantes à travers les pratiques honorant les ancêtres. Dans la première partie, nous nous intéresserons aux formes que prend le culte des ancêtres au sein de la maisonnée et entre les lignées ainsi qu’aux caractéristiques de la tablette paqweil. Cette 366 dernière constitue un vecteur entre les membres de la maisonnée et ses ancêtres patrilinéaires. Puis, plus brièvement, le processus du deuil sera abordé à partir d’une description des rites funéraires, en discutant la « naissance » d’un ancêtre. Par cette approche, ce chapitre questionne non seulement les modalités de culte des ancêtres mais aussi le rôle et la place des femmes, que ce soit comme officiantes ou comme ancêtres. I. Le culte des ancêtres Comme dans beaucoup de sociétés d'Asie, le culte des ancêtres est central en Chine. Il entre dans la catégorie de la « religion de lignage » discutée par Joël Thoraval (1991). Le culte des ancêtres tient également une place prépondérante au sein de la vie villageoise de Wumu. Comme le précise Jacques Lemoine (1973 : 147) dans un article sur le culte des ancêtres chez les Hmong (Laos), « le culte des Ancêtres s'analyse en fait comme un culte de la lignée, c'està-dire de l'ensemble des âmes dans un groupe de commune descendance [...] ». Il est vrai qu'à Wumu, le culte des ancêtres s'accomplit notamment au niveau de la lignée. Le culte des ancêtres de la ligne directe, effectué quotidiennement au sein d'une maisonnée, est la forme la plus fréquente, on l'observe à l'occasion des rituels quotidiens d'offrandes de thé et de nourriture, ainsi que lors du réveillon du Nouvel An. Tous les premier et quinzième jours de chaque mois, les offrandes entourant la tablette sont renouvelées. Le culte des ancêtres est par ailleurs accompli à divers autres niveaux. À l'échelle d’un groupe de lignées, directes et collatérales conjointes, le culte des ancêtres est accompli ponctuellement ou annuellement. Les ancêtres sont honorés lors d'événements significatifs, en hiver lors des visites des vœux du nouvel an lunaire et du nettoyage des tombes, pendant le second mois lunaire à l'occasion des célébrations des 36 ans et des 49 ans des hommes du village, au printemps pour la fête des morts lors de la visite des tombes et pour le premier jour du sixième mois lunaire, nommé chual-mei tal-biuq, le « culte estival aux ancêtres du sixième mois ». Enfin, l’envergure du Sacrifice aux ancêtres maternels célestes, l'une des deux grandes cérémonies collectives auxquelles participe annuellement le village entier, dépasse les simples pratiques de la vie quotidienne de culte des ancêtres car il s'inscrit dans un élan régional de renouveau du patrimoine culturel naxi et puisque les institutions culturelles et le gouvernement local interviennent dans son organisation. Sa performance se distingue difficilement de l'aspect de mise en scène qui caractérise tant de pratiques rituelles soutenues et développées par les autorités officielles dans le cadre des politiques des minorités en Chine. 367 Le culte des ancêtres el-cif-lol-el-cif-nail et epvq-esse par les habitants de Wumu imprègne le quotidien et est intégré à toutes les pratiques rituelles naxi de même qu'à toute festivité plus ou moins importante impliquant la tenue d'un banquet. Quelles qu’en soient l’occasion et la fréquence, le dialogue avec les ancêtres est assuré en toutes circonstances, au quotidien comme pour les événements extraordinaires. C'est un processus ordinaire porté dans tous les aspects de la vie villageoise, qui, bien qu'associé à des performances diverses, suit en général un même mode opératoire. À Wumu, il est souvent associé à des petits rites naxi renvoyant aux divinités ou esprits malfaisants relevant de la pratique et des mythes dongba. A. Ancêtres et tablettes Les ancêtres epvq-esse sont omniprésents dans la vie et les activités quotidiennes des villageois. La cuisine « yagoq » dans la maison « yagoq » (voir Chapitre 6) est la pièce centrale dans la performance rituelle du culte aux ancêtres de la maisonnée, où les ancêtres sont représentés matériellement par une tablette. Il s'agit d'une pièce de bois d'environ trente centimètres de haut sur laquelle sont inscrits, en chinois, les noms et dates de naissance et décès des défunts de la famille. Elle est placée en hauteur sur un meuble ou sur une planche où des offrandes sont déposées quotidiennement. La tablette doit être placée dos au mur, à l'Est, et faisant face à l’Ouest. Appelée communément paqweil par les habitants du village, adaptation en naxi du terme chinois panwei ( 盘位 ), cette tablette retrace les ancêtres respectifs de la maisonnée sur trois générations, suivant le cours des lignées, à partir des parents décédés le plus récemment dans la maison. Les ancêtres epvq-esse, el-cif-lol et el-cif-nail sont des membres à part entière de la maisonnée, les offrandes à leur égard sont renouvelées régulièrement et leurs descendants les honorent avant chaque repas. Ces ancêtres prennent part à chaque grand événement important ayant lieu dans l'enceinte de la maison. Si trois générations d'ancêtres figurent déjà sur la tablette lorsqu'un nouveau décès intervient, elle est alors remplacée et brûlée dans le foyer de la cuisine, et le nom du défunt doit être inscrit sur une nouvelle tablette vierge, qui ne comporte alors plus que les noms d’une seule génération. La continuité de la filiation est essentielle, elle doit suivre son cours et ne peut sauter de générations. Ainsi, dans la cuisine du vieux A-De, un doyen des Miq we jjiq, l'étagère rituelle porte non pas une mais deux tablettes des ancêtres. Une pour son père, sa mère et son grandpère paternel, une seconde pour ses deux fils décédés prématurément quelques années auparavant. L'une des règles fondamentales du deuil à Wumu est qu'il n'est pas possible de 368 porter le deuil et d’honorer comme ancêtre un parent plus jeune que soi en génération et en âge. De fait, tant qu’A-De et son épouse sont encore vivants, les noms de ses ancêtres et ceux de leurs deux fils ne peuvent être tous inscrits sur une seule et même tablette. De même, au cimetière, les tombes des deux fils d’A-De ne peuvent être identifiées par une stèle gravée tant qu’A-De et son épouse sont encore vivants. Certains doyens du village ont disposé d'autres figurines aux côtés de la tablette des ancêtres, au milieu des offrandes de nourriture, des bâtons d'encens et des vases, comme une statuette de Mao remontant à l'époque de la Révolution Culturelle (1966-1976) ou une figurine de Guanyin, selon les croyances des familles. Sous le régime de Mao Zedong, il était interdit de placer la tablette des ancêtres dans la cuisine et quiconque contrevenait à cette règle pouvait être exécuté, selon les dires d’A-Yu. Ce dernier se souvient encore de cette époque-là, la fin de son adolescence, et invoque le combat de Mao contre les Quatre Vieilleries89 (po sijiu 破四旧) et contre les superstitions (mixin 迷信). Pendant toute cette période, les tablettes ont été retirées des cuisines. La tablette traduit deux dynamiques, l'une est l’action d’honorer les ancêtres de la maisonnée, elle représente le devoir de piété filiale envers les ascendants par les descendants. L'autre dynamique est la continuation du processus de filiation. L'anthropologue Maurice Freedman parlait dans Family and Kinship in Chinese Society (1970) de la chaîne de filiation (« Continuum of Descent »), dans laquelle ancêtres et descendants dépendent mutuellement les uns des autres. La maisonnée constitue un maillon de cette chaîne, le lien entre le réseau ancestral et le réseau social de parenté. La tablette fournit un intermédiaire entre ces deux réseaux, un entre-deux matériel. Elle ancre également tout individu à son yagoq, sa maison. Toutefois, il n’y a de tablette dans une nouvelle maison – généralement issue d'une séparation du foyer – que si une personne y est décédée. Un homme du village m'a un jour expliqué qu'étant le plus jeune de sa fratrie, il a pu construire sa propre maison indépendante. Il y a emménagé avec son épouse, ils y vivent avec leurs deux enfants. Ils n'ont cependant pas de tablette des ancêtres dans la maison, ce qu'il m'a expliqué par le fait que personne n'est encore mort sous son toit. Au moment des repas, en l'absence de tablette dans la cuisine, c'est donc en 89 Les Quatre Vieilleries, objets de la campagne de destruction de Mao Zedong pendant la Révolution Culturelle (1966-1976), sont les « vieilles idées » (jiu sixiang 旧思想), la « vieille culture » (jiu wenhua 旧文化), les « vieilles coutumes » (jiu fengsu 旧风俗) et les « vieilles habitudes » (jiu xiguan 旧习惯). 369 direction de sa maison de naissance, dont il est parti par division du foyer, qu'il doit présenter les plats et honorer les ancêtres. B. Transfert de tablettes et continuité de culte En juillet 2018, A-You, une femme veuve, est décédée, laissant derrière elle quatre descendants : ses deux fils et ses deux neveux, les fils du frère de son mari (voir Figure 26). J’identifie ici lesdits neveux comme des descendants car, nous allons le voir, ils ont pris part aux rites funéraires au même titre que les fils, en portant eux-aussi le chapeau de descendant (Photographie 30). Cela nous questionne : se pourrait-il que l’objet du deuil ne soit pas seulement la défunte, mais également le dernier doyen de toute une lignée? Se pourrait-il encore que les descendants ne fassent pas le deuil d’une seule personne, mais du couple ancestral – à présent réuni – et de son lien à la « maison-mère »? Ces funérailles auraient-elles été un point de jonction deux générations, dans la continuité de la chaîne de filiation? Comme nous le verrons plus loin, les funérailles se déroulent comme de coutume au village, pendant trois jours. Les rites funéraires permettent à un défunt d'accéder au statut d'ancêtre, et, le deuxième jour, son nom est inscrit au côté des noms des ancêtres sur la tablette des ancêtres de la lignée. Lors de funérailles d'un membre de la maisonnée, la tablette paqweil est sortie de la cuisine et posée sur une table d'offrandes dressée dans la cour de la maison, pour y être complétée du nom du défunt par un homme lettré choisi par la famille. L'été 2018, la tablette a donc été sortie de la cuisine. Le nom et les dates de naissance et de décès de la défunte A-You y ont été inscrits par un voisin lettré, ancien maître d’école, à la suite de ceux des ancêtres de la maisonnée. Mais, à ma grande surprise, une seconde tablette de bois a été fabriquée, et reproduite à l'identique de la première tablette. A-You a en effet deux fils, le plus jeune ayant hérité de la maison et prenant soin de sa mère, l'aîné ayant séparé son foyer à son mariage pour emménager de l’autre côté de la rue dans sa propre maison avec son épouse et ses enfants. Aux funérailles de leur mère, la première tablette a retrouvé sa place dans la cuisine de la maison familiale, la seconde tablette a, elle, été portée chez le fils aîné de l'autre côté de la ruelle. C'est ici la seule occasion où il m'a été donné d'observer une paqweil sortir de l'enceinte de la maison. Plus exactement, l’inscription de la nouvelle tablette marque le moment où une maison issue d’une séparation de l’os (o-bbiu) et jusqu’à présent sans tablette, 370 « intègre » son premier ancêtre. A-You vivait avec son fils cadet, cependant son fils aîné pourra également l’honorer par des offrandes quotidiennes dans sa propre maison.	13,598
https://openalex.org/W2972341748	OpenAlex	Open Science	CC-By	2,021	Quantum-inspired algorithms for multivariate analysis: from interpolation to partial differential equations	Juan José García‐Ripoll	English	Spoken	18,477	34,517	Quantum-inspired algorithms for multivariate analy- sis: from interpolation to partial diﬀerential equations Juan José García Ripoll Institute of Fundamental Physics, Calle Serrano 113b, 28006 Madrid, Spain In this work we study the encoding of smooth, diﬀerentiable multivariate function in quantum registers, using quantum computers or tensor-network representations. We show that a large family of distributions can be encoded as low-entanglement states of the quantum register. These states can be ef- ﬁciently created in a quantum computer, but they are also eﬃciently stored, manipulated and probed using Matrix-Product States techniques. Inspired by this idea, we present eight quantum-inspired numerical analysis algorithms, that include Fourier sampling, interpolation, diﬀerentiation and integration of partial derivative equations. These algorithms combine classical ideas—ﬁnite- diﬀerences, spectral methods—with the eﬃcient encoding of quantum registers, and well known algorithms, such as the Quantum Fourier Transform. When these heuristic methods work, they provide an exponential speed-up over other classical algorithms, such as Monte Carlo integration, ﬁnite-diﬀerence and fast Fourier transforms (FFT). But even when they don’t, some of these algorithms can be translated back to a quantum computer to implement a similar task. v:1909.06619v5 [quant-ph] 12 Apr 2021 arXiv:1909.06619v5 [quant-ph] 12 Apr 2 arXiv:1909.06619v5 [quant-ph] Juan José García Ripoll: jj.garcia.ripoll@csic.es, http://quinfog.hbar.es Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 1 Introduction Quantum computers use the exponential capacity of a Hilbert space to process information. A quantum computer with m qubits can store 2m complex numbers as the components of the quantum register wavefunction, \|ψ⟩= P2m−1 s=0 ψs \|s⟩. A quantum algorithm creates, manipulates and probes these amplitudes to solve a concrete problem. There exist al- gorithms where the amplitudes ψ(s) encode a smooth function deﬁned over a volume in RN. A common example is storing probability distributions in the quantum register [1, 2] and developing algorithms to extract expected values [3] or conditional probabilities [4]. With this, it becomes possible to perform valuations of complex ﬁnancial assets [5, 6], VaR estimates [7], and other sophisticated interrogations. Using a similar encoding, one may also address radically diﬀerent problems, such as solving partial diﬀerential equations with ﬁnite diﬀerences [8–10] or variational methods [11]. In this work we discuss how eﬃcient it is to encode discretized functions in a quan- tum register. We ﬁnd that for certain distributions—smooth diﬀerentiable functions with bounded derivatives or bounded spectrum—, the accuracy of the discretization increases exponentially with the number of qubits, while the bipartite entanglement grows slowly or even remains bounded with the problem size. This implies such distributions may be constructed with quasi-local operations and polynomial resources on quantum com- puters. But it also opens an exciting possibility: a family of quantum-inspired matrix- product state (MPS) techniques that can approximate the low entanglement state of a Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 1 quantum register and provide new classical (and quantum) algorithms for interpolating, diﬀerentiating, Fourier transforming or solving diﬀerential equations of such distributions. These techniques work eﬃciently because of an implicit renormalization where diﬀerent qubits work with diﬀerent length scales, in a way that lends itself to eﬃcient interpolation and compression. This idea, analogous to multigrid renormalization method developed in Ref. [12], has strong parallelisms to earlier developments in the 2D quantum image pro- cessing world [13], gives rise to a performance improvement over earlier techniques based on tensor trains [14, 15] or MPS encodings of mode expansions [16]. This paper is structured in three parts. The bulk of the work is preceded by a summary (Section 2) of the main results and heuristic algorithms that are developed in this work. The ﬁrst section addresses the representation of discretized functions in quantum registers. It presents state-of-the-art techniques (Sect. 1 Introduction 3.1) and new discretizations (Sect. 3.2) on an equal footing. We argue that these samplings produce to weakly entangled multi-qubit states (Sect. 3.3). This prediction is conﬁrmed numerically for common distributions in 1, 2 and 3 dimensions, using exact simulations of up to 28 qubits. The second part of this work builds on this result, introducing the idea of using matrix-product states (MPS) for representing the low-entanglement encoding of such functions in quantum registers. In other words, we propose using an MPS-encoded (classically simulated) quantum register to store, compute, manipulate and interrogate smooth multivariate functions. Section 4 articulates this idea, recalling well known algorithms from the literature and how they specialize for our purposes. With these tools at hand, Section 5 develops new quantum- inspired algorithms for the numerical analysis of multivariate functions. These include the mapping of functions to MPS format (Sect. 5.5), Fourier analysis (Sect. 5.1), interpolation methods (Sect. 5.2), and techniques for approximating derivatives of discretized functions (Sect. 5.3), both through ﬁnite-diﬀerence (Sect. 5.3.1) and the Quantum Fourier Transform (QFT, Sect. 5.3.2). Sect. 5.4 combines these techniques into higher level algorithms for solving partial derivative equations, demonstrating their performance in the Fokker-Planck model. This work is closed with a discussion of the results, including connections to recent advances in tensor-based numerical analysis, and an outlook of applications. 2 Summary Quantum computing has introduced the clever idea of encoding probability distributions in quantum registers. In this encoding, a small number of qubits can store an exponen- tially large number of function samples. To ﬁx ideas and notation, let us take a function p(x1, . . . , xN) of N variables in a bounded interval. Let us use m qubits to uniformly discretize the domain xi s = ai + bi −ai 2m si = ai + δi m m X k=1 sk i 2k , i = 1, 2 . . . N. (1) (1) The non-negative integer si takes all possible values obtained by grouping m bits s1 i s2 i · · · sm i , ordered in decreasing signiﬁcance. The N integers or Nm bits can be associated to diﬀer- ent states of a quantum register, enabling two representations of the function p. The ﬁrst one assumes a non-negative function, The non-negative integer si takes all possible values obtained by grouping m bits s1 i s2 i · · · sm i , ordered in decreasing signiﬁcance. The N integers or Nm bits can be associated to diﬀer- ent states of a quantum register, enabling two representations of the function p. The ﬁrst one assumes a non-negative function, \|p⟩∝ X s1,...,sN q p(s1, . . . , sN) \|s1⟩⊗· · · ⊗\|sN⟩, si ∈{0, 1, . . . 2m −1}, (2) (2) Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 2 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 2 Problem Algorithm Type Cost Expected value Monte Carlo C O(1/ε2) [3] " Amplitude estimation Q O(1/ε) [3] " MPS QI O(Nχ3 log2(1/ε)) [cf. §4.3] Fourier transform QFT Q O(Nm log(Nm)) [17] " FFT C O(Nm2Nm) [18] " MPS QFT QI O(Nm × SimpNm) [cf. §5.1] Interpolation Linear (k = 1) C O(2Nm) [18] " MPS Linear (k = 1) QI ∼SimpNm [cf. §5.2.1] " FFT C O(N(m + k)2N(m+k)) [cf. §5.2.2] " MPS QFT QI ∼QFTN(m+k) [cf. §5.2.2] PDE Evolution Finite diﬀerences C O(Tcgs22Nm) [18] " MPS diﬀerences QI O(Tcgs × SimpNm) [cf. §5.4.1] " FFT method C O((Nm + 1)2Nm) [18, 19] " MPS QFT QI ∼2 × QFTN(m+k) [cf. §5.4.2] State construct GR-like Q O(Nmχ2) [cf. §3.3] " Explicit wavefunction C O(2Nm) " MPS QI O(Tsteps × SimpNm) [e.g. 2 Summary 3 carry information over long wavelengths and large features, and the least signiﬁcant bits sm 1 , sm 2 . . . are eﬃciently approximated with low-entanglement interpolation methods over those qubits (Sect. 5.2). These ﬁndings suggest that many useful functions and problems can be constructed in a quantum register with polynomial resources, due to the bounded entanglement (see Sect. 3.3). However, the same results open the ﬁeld of quantum-inspired numerical anal- ysis, which combines the quantum computing encoding of functions with tensor-network representations and algorithms to manipulate them (see Sect. 4). This approach sug- gests new algorithms for integrating probability distributions and computing expected values (Sect. 4.3), for implementing discrete diﬀerentiation (Sect. 5.3), Fourier transform (Sect. 5.1), interpolation (Sect. 5.2) and for the solution of partial diﬀerential equations (Sect. 5.4). Table 1 summarizes the algorithms discussed in this work, focusing on ﬁve diﬀerent problems (i) computation of expectation variables from probability distributions, (ii) con- struction of probability distributions (see Section 5.5), (iii) Fourier analysis (Section 5.1) and (iv) interpolation of discretized functions (Section 5.2, and (v) solution of partial dif- ferential equations (Section 5.4). The table pairs the quantum-inspired methods based on matrix-product states (type QI), with alternatives that already exist for quantum com- puters (type Q) or in the ﬁeld of classical numerical analysis (type C). For each method, the table provides an estimate of the algorithm’s cost, expressed in terms of well known quantities: desired error bounds ϵ, number of dimensions of the functions involved N, di- mension d = 2 of quantum register elements, logarithm of the discretization size along each dimension m, estimated bond dimension χ for the MPS representation, number of steps in the MPS approximation algorithms Tstep, Tsweeps. In the quantum and classical versions, we provide versions of the bounds that are found in the literature, specialized for the parameters we have just mentioned, together with original of state-of-the-art references. In the case of quantum-inspired numerical analysis, we provide standard heuristic perfor- mance metrics based on the literature [20, 21]. These estimates use parameters, such as the bond dimension χ or the number of iterations in the approximation algorithm Tsweeps, that are very much problem-dependent. 2 Summary §4.5] MPS algorithms Simpliﬁcation (SimpNm) C O(TsweepsNm4d3χ3) [20] " Expected values C O(Nm × 2dχ3) [20] " MPO-MPS product C O(Nm(dχχf)2) [20] Table 1: Algorithms and their costs. We compare the costs of diﬀerent tasks when working with multivariate functions, from the construction of the state, to the simulation of their evolution. We use the following heuristic values: ε, desired error bound; N, number of variables; m ∼O(log2(1/ε)), number of qubits per variable for 2m points in discretization; d = 2, physical dimension of qubits; χ, χf, eﬀective MPS and MPO bond dimensions; Tsweeps number of iterations in the simpliﬁcation algorithm (cf. Sect. 4.4); Tcgs, number of iterations in conjugate gradient method (cf. Sect. 5.4.1). Some of the algorithms give rise to their own heuristic metrics: SimpK, cost of simplifying an MPS with K qubits; QFTK, cost of simulating a QFT using MPS with K qubits. We distinguish classical methods (C), from algorithms for a quantum computer (Q), or classical MPS techniques that imitate the quantum register approach (QI). We also reference the papers or sections in this work where such costs are discussed. and associates p(s) to the probability of the state \|s⟩. The second encoding does not make this assumption and maps the distribution directly to the wavefunction \|p⟩∝ X s1,...,sN p(s1, . . . , sN) \|s1⟩⊗· · · ⊗\|sN⟩. (3) (3) Both representations can be extended to situations where the functions are not normalized, just by keeping track of global prefactors. In this work we argue that both representations are exponentially eﬃcient in many ways. First, the quantum register demands only a logarithmically growing number of qubits Nm to store an exponential amount of weights 2Nm in a discretized function. Sec- ond, we also need an exponentially small number of qubits m ∼−log(ε) to reduce the discretization error below a given tolerance ε. Third and ﬁnally, we ﬁnd that for smooth, diﬀerentiable functions with bounded derivatives, these states have a small amount of en- tanglement. Indeed, for many distributions of interest we obtain an entanglement scaling O(N) with the dimension of the problem. We conjecture that this behavior is due to an implicit renormalization that happens in the quantum register, where some bits s1 1, s1 2 . . . 3 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 2 Summary However, as we will see throughout this work, if entanglement remains bounded throughout the simulations, the MPS-simulated quantum register demands a small bond dimension χ, and the algorithms provide an exponential speedup over other classical techniques—from ﬁnite diﬀerences to the highly performant FFT techniques. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 3.1 GR discretization One of the earliest works suggesting the encoding of functions in quantum registers is the work by C. Zalka [1], later rediscovered by Grover and Rudolph [2]. This designed a unitary operator Up that encodes a probability distribution p(x) in an empty quantum register with m qubits \|p(m) GR ⟩:= Up \|0, 0, . . . , 0⟩= 2m−1 X i=0 q p(m) GR (s) \|s⟩, with p(m) GR (s) = Z xs+1 xs p(u)du. (4) (4) The original construct assumes a random variable x in a bounded interval [a, b] subdivided into 2m smaller intervals, labeled by the quantum register states \|s⟩= \|s1s2 · · · sm⟩. The original construct assumes a random variable x in a bounded interval [a, b] subdivided into 2m smaller intervals, labeled by the quantum register states \|s⟩= \|s1s2 · · · sm⟩. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 4 a) c) b) Figure 1: (a) A probability state \|p(m)⟩with discretization size 2m+1 can be constructed from a coarser state \|p(m)⟩by appending one auxiliary qubit through a unitary operation U (m). (b) When a state of m qubits is upgraded to m + k, we can study the entanglement between the old and new qubits through a Schmidt decomposition (14). (c) If the state has a small Schmidt number for all 1D bipartitions, it is weakly entangled and admits an eﬃcient MPS representation (29). a) c) b) b) a) c) Figure 1: (a) A probability state \|p(m)⟩with discretization size 2m+1 can be constructed from a coarser state \|p(m)⟩by appending one auxiliary qubit through a unitary operation U (m). (b) When a state of m qubits is upgraded to m + k, we can study the entanglement between the old and new qubits through a Schmidt decomposition (14). (c) If the state has a small Schmidt number for all 1D bipartitions, it is weakly entangled and admits an eﬃcient MPS representation (29). A practical application of this encoded state would be the computation of expected values for any observable or function f(x). This requires engineering an observable ˆO(m) f such that Z A practical application of this encoded state would be the computation of expected values for any observable or function f(x). This requires engineering an observable ˆO(m) f such that ¯f = Z f(x)p(x)dx ≃⟨p(m) GR \| ˆO(m) f \|p(m) GR ⟩+ εint. 3.1 GR discretization (5) (5) Typically, we approximate ˆO(m) f = P s f(xs) \|s⟩⟨s\| , and apply a uniform discretization (1), to have an integration error that decays exponentially with register size, εint ∼O(δm). As found by A. Montanaro [3], using amplitude estimation with Up and the operator ˆOf, one may estimate ¯f with a precision that scales better than Monte Carlo algorithms. If the cost of implementing Up is TGR, and we aim for a sampling precision εsample, the asymptotic time cost of the ideal amplitude estimation algorithm is roughly TQCMC = O (TGR/εsample) . (6) (6) This represents a favorable scaling when compared with traditional Monte Carlo, where the sampling uncertainty goes as O(ε−2 sample), but only if the cost of encoding the probability state TGR remains small or weakly dependent on the integration error εint. The Up is a recursive construct that adds one more qubit of precision in each step. As sketched in Fig. 1a, the procedure reads \|p(m+1) GR ⟩= u(m+1) \|p(m) GR (s)⟩\|0⟩= 2m−1 X s=0 q p(m) GR (s) \|s⟩(cos(θs) \|0⟩+ sin(θs) \|1⟩) (7) (7) = 2m+1−1 X s′=0 q p(m+1) GR (s′) \|s′⟩, = 2m+1−1 X s′=0 q p(m+1) GR (s′) \|s′⟩, where we identify \|s⟩\|0⟩=: \|2s⟩and \|s⟩\|1⟩=: \|2s + 1⟩. The rotation angle 0 ≤θi ≤π/2 is obtained from two identities where we identify \|s⟩\|0⟩=: \|2s⟩and \|s⟩\|1⟩=: \|2s + 1⟩. The rotation angle 0 ≤θi ≤π/2 is obtained from two identities where we identify \|s⟩\|0⟩=: \|2s⟩and \|s⟩\|1⟩=: \|2s + 1⟩. The rotation angle 0 ≤θi ≤π/2 obtained from two identities cos(θs)2 = p(m+1) GR (2s) p(m) GR (s) , sin(θs)2 = p(m+1) GR (2s) p(m) GR (s) . (8) (8) Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5 Unfortunately, this algorithm requires an exponentially large number of angles and involves a highly non-local unitary with a potentially bad decomposition. The GR algorithm must be therefore considered more a proof of existence, than a practical recipe that can be used when bounding the resources of Monte Carlo analysis. We will study the GR construct and other discretizations, demonstrating that there are eﬃcient alternatives to equation (7). Our analysis centers on the complexity of the sampled states. Using the bipartite entanglement as quantiﬁer, we will show that the cost of adding one more qubit of resolution decays exponentially. 3.1 GR discretization This will help us understand that there one quasi-local unitary procedure that builds the GR state with a cost that is polynomial in the number of qubits, TGR ∼O(m log2(1/ε)). We will conﬁrm numerically this result using various well-known probability distributions. 3.2 Other discretizations The integral representation (4) encodes the probability that is contained inside each inter- val, but it requires computing 2m integrals. In practice, this is unnecessary because the estimation of expected values already introduces a discretization error (5) εsample ∼O(δm) in the operator deﬁnition. It is not diﬃcult to ﬁnd simpler representations that have the same or better scaling. The obvious one is the uniform sampling of the probability distribution \|p(m) R ⟩= s δm Nm X s q p(xs) \|s⟩, with Nm = X s δmp(xs). (9) (9) The standard error bound for this Riemann-type state is εsample ≤max d dx(fp) δm, which depends on the derivatives of both the sampled observables and probability. The ﬁrst order scaling is good enough for the simulations that we will show below, because the interval size decreases exponentially with the number of qubits m. We will therefore stick to the GR states or to Eq. (9), unless otherwise noted. However, if we need to save some qubits, we can try variations, such as a probability state that implements the trapezoidal or the Simpson rule \|p(m) S ⟩= 1 S1/2 m X s q θsp(xs) \|s⟩, θs = ( 1, s = 0, 2m −1 4 −(s mod 2), else. (10) (10) The discretization error of the Simpson state decreases faster with the interval size, εsample ∼ O(δ2 m). This scaling means that the qubits required to achieve a given precision m ∼ log2(1/εsample) can be half those required by the uniform ansatz. These savings may be interesting in resource-starved architectures, such as NISQ computers, and also in the algorithms to be considered later in Sect. 4. 3.3 Entanglement bounds We can show that adding k new qubits to a one-dimensional GR probability state with m qubits demands a vanishingly small amount of entanglement, that decreases exponentially with 2m. This small amount of entanglement suggests that the GR unitary U(m) p can be replaced with simpler decomposition in terms of quasi-local gates. The reasoning proceeds as follows. Assuming that the probability distribution is smooth, diﬀerentiable and has an upper bound on its derivative, Dp = max x \|p′(x)\|, (11) (11) 6 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. we can obtain upper bounds for the entropy of the extra qubit. For one added qubit, k = 1 our bound reads q q S[ρ(m,1)] ≤ q 2(1 −P) ≤2 q Dp\|b −a\|2−m/2. (12) (12) If k is larger, we have roughly If k is larger, we have roughly S[ρ(m,k)] ≤O 2 q Dp\|b −a\|2−(m−k)/2 . (13) (13) Note that this argument easily extends to the Riemann-type (9) or Simpson sampling (10), because the diﬀerences between them decays exponentially with the number of qubits. To prove these bounds, assume that we have produced a GR state with m qubits and we wish to extend it to include k additional qubits. Our goal is to show that there exists an Schmidt decomposition of the new state into two sets of m and k qubits \|p(m+k)⟩= X α λ1/2 α \|ξ(m) α ⟩⊗\|ξ(m,k) α ⟩, (14) (14) with a small amount of entanglement between its components S[ρ(m,k)] = − X α λα log(λα). (15) (15) Our ﬁrst step is to derive an expression for ρ(m,k). In our extended state, the coordinates x are a function two numbers of m and k bits, which we label s and u. These numbers combine to form a larger previous index s with m + k bits. More precisely Our ﬁrst step is to derive an expression for ρ(m,k). In our extended state, the coordinates x are a function two numbers of m and k bits, which we label s and u. These numbers combine to form a larger previous index s with m + k bits. More precisely x ∈a + δ(m)s + δ(m+k)u =: rs + ϵu, ( s = 0, 1, . . . , 2m −1, u = 0, 1, . . . , 2k −1. 3.3 Entanglement bounds In the particular case in which k = 1, the two eigenvalues of the reduced density matrix are λ± = 1 2 1 ± √ 2P −1 . (25) (25) Using the bound H2(x) = −x log2(x) −(1 −x) log2(1 −x) ≤2 q x(1 −x), (26) H2(x) = −x log2(x) −(1 −x) log2(1 −x) ≤2 q x(1 −x), (26) (26) we ﬁnd that the von Neumann entropy is upper bounded by Eq. (12). We can also bound the von Neumann entropy of the reduced density matrix for a larger number of extra qubits. Introducing the trace norm of σ We can also bound the von Neumann entropy of the reduced density matrix for a large number of extra qubits. Introducing the trace norm of σ We can also bound the von Neumann entropy of the red number of extra qubits. Introducing the trace norm of σ T[σ] = 1 2∥σ∥1 = X uv \|σuv\| ≤Dp(b −a)2 1 2m−k+1 . (27) (27) Manipulating the Fannes-Audenaert bound [22] Manipulating the Fannes-Audenaert bound [22] \|S(ρ) −S(\|e⟩⟨e\|)\| ≤T log2(2k −1) + H2(T) ≤T(k −e−k+1) + 2 √ T, (28) (28) we have that the entropy can be made arbitrarily close to zero as we make T smaller and smaller, by increasing 2mc−k+1. For suﬃciently small T, we can neglect the ﬁrst term and write (12). 3.3 Entanglement bounds (16) (16) For convenience, we will us split xz = rs + ϵu, separating a large and small length scale. We can write q ce, we will us split xz = rs + ϵu, separating a large and small length scale. For convenience, we will us split xz = rs + ϵu, separating a large and small length scale. We can write q We can write ρ(m,k) = X s,u,v q p(m+k)(rs + ϵu)p(m+k)(rs + ϵv) \|u⟩⟨v\| . (17) (17) Starting with Eq. (17), we split the density matrix into Starting with Eq. (17), we split the density matrix into ρ(m,k) = \|e⟩⟨e\| + σ, (18) (18) ρ \| ⟩⟨\| , ( ) with the unit vector \|e⟩= 1 2k/2 2k−1 X u=0 \|u⟩, (19) with the unit vector \|e⟩= 1 2k/2 2k−1 X u=0 \|u⟩, (19) (19) and the perturbation and the perturbation ⟨u\|σ\|v⟩= X s q p(m+k)(rs + ϵu)p(m+k)(rs + ϵv) −1 2k p(m)(rs) . (20) (20) Thanks to the mean-value theorem, we know that p(x) achieves its mean value Thanks to the mean-value theorem, we know that p(x) achieves its mean value ¯p(m) = 1 δ(m) p(m)(rs) = p(¯rs) (21) (21) at some point ¯rs in the interval [rs, rs + δ(m)]. We can therefore bound at some point ¯rs in the interval [rs, rs + δ(m)]. We can therefore bound p(m+k)(rs + ϵu) = Z rs+ϵu+δ(m+k) rs+ϵu p(w)dw ≤δ(m+k) × h p(¯rs) + Dpδ(m)i , (22) (22) Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. where Dp = maxa≤x≤b p′(x). Using δ(m+k) = δ(m)/2k, we bound where Dp = maxa≤x≤b p′(x). Using δ(m+k) = δ(m)/2k, we bound \|σ \| ≤ X 1 D (δ(m))2 D (b a)2 1 (23) where Dp = maxa≤x≤b p′(x). Using δ(m+k) = δ(m)/2k, we bound ≤b p′(x). Using δ(m+k) = δ(m)/2k, we bound \|σuv\| ≤ X s 1 2k Dp(δ(m))2 = Dp(b −a)2 1 2m+k (23) (23) We can now use this bound in various ways. We can for instance study the purity of the state, using tr(σ2) ≥0 to obtain We can now use this bound in various ways. We can for instance study the purity of the state, using tr(σ2) ≥0 to obtain P[ρ] = trρ2 = 1 + tr(σ2) −2tr ⟨e\|σ\|e⟩≥1 −2Dp(b −a)2 1 2m , (24) (24) a value that vanishes exponentially with the number of qubits. 3.4 MPS representation 2 4 6 8 10 12 m 10 5 10 2 10 1 b) 2 4 6 8 10 12 m 0.00 0.25 0.50 0.75 1.00 S[ (m, 1)] a) Gaussian Log-normal Lorentzian Non-convex 2 4 6 8 10 12 k 10 5 10 3 10 1 S[ (m k, k)] c) Gaussian Log-normal Lorentzian Non-convex 0 1 2 3 4 5 0.5 1.0 max(S[ (M k, k)]) d) c) d) Figure 2: (a) Entropy of entanglement between the m ﬁrst qubits and an additional qubit that is added to a GR state. We use (σ, µ) = (1, 0), (1, 1), (1, 0), (1, 0) for the Gaussian, Log-normal, Lorentzian and non-convex distributions, choosing the intervals [a, b] = [µ −6σ, µ + 6σ], [10−16, µ + 50σ], [µ − 10σ, mu + 10σ] and [0, 7σ]. (c-d) Similar as (a), but we plot the entanglement of a bipartition with (k, M −k) qubits, for M = 14. are pictures produce a macroscopic number of extremely small eigenvalues in any Schmidt decomposition. These eigenvalues are too small to make the purity or the von Neumann entropy grow, but are too many for an accurate and eﬃcient MPS representation of the state. However, as we will see below with practical simulations, this does not happen for many smooth functions. Instead, we ﬁnd very accurate representations of smooth func- tions in 1, 2 an 3 dimensions using very small MPS. This has important consequences for both quantum and classical algorithms. First, MPS states have eﬃcient sequential con- structs [24, 25] with bounded time-cost TGR ∼O(mχ2), potentially smaller than the GR construct. Such constructs are very relevant for the implementation of quantum numer- ical analysis in quantum computers. Second, MPS states can be manipulated eﬃciently in classical computers, leading to a new family of quantum-inspired numerical algorithms introduced in later sections. 3.4 MPS representation The fact that adding every new bit requires a small amount of entanglement suggests that the probability state \|p(m)⟩has an eﬃcient matrix-product state (MPS) representation. A matrix product state is a decomposition of a wavefunction as a contraction of matrices that are labeled by the physical indices of a composite quantum system \|p(m)⟩= X s q p(m)(s) \|s⟩≃ X As1 α1As2 α1,α2 · · · Asm αm \|s1, s2, . . . , sm⟩. (29) (29) Each of the tensors is in general diﬀerent, and has three indices Asi αi,αi+1 ∈C2×χi×χi+1. There is one physical index s of dimension 2, and two bond dimensions of sizes χi and χi+1. Interestingly, MPS states with a small bond dimension χi have also small von Neumann entropy, S[ρ(i,n−i)] ≤O(χi). Heuristically, we could argue that the bounds for smooth functions indicate the repre- sentability of such functions using weakly entangled MP states. In full rigor, the bounds we obtained are bounds on Renyi entropies Sα with α >= 1. This means that, as shown by N. Schuch et al [23], the MPS simulability of the states is undetermined. This ap- parent contradiction is observed in the MPS encoding of images [13]. In this case, there Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 8 2 4 6 8 10 12 m 0.00 0.25 0.50 0.75 1.00 S[ (m, 1)] a) Gaussian Log-normal Lorentzian Non-convex 2 4 6 8 10 12 m 10 5 10 2 10 1 b) 2 4 6 8 10 12 k 10 5 10 3 10 1 S[ (m k, k)] c) Gaussian Log-normal Lorentzian Non-convex 0 1 2 3 4 5 0.5 1.0 max(S[ (M k, k)]) d) Figure 2: (a) Entropy of entanglement between the m ﬁrst qubits and an additional qubit that is added to a GR state. We use (σ, µ) = (1, 0), (1, 1), (1, 0), (1, 0) for the Gaussian, Log-normal, Lorentzian and non-convex distributions, choosing the intervals [a, b] = [µ −6σ, µ + 6σ], [10−16, µ + 50σ], [µ − 10σ, mu + 10σ] and [0, 7σ]. (c-d) Similar as (a), but we plot the entanglement of a bipartition with (k, M −k) qubits, for M = 14. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 3.5 Numerical study of 1D distributions Let us put these ideas to the test using three paradigmatic distributions Gaussian: pG(x; σ, µ) = 1 √ 2πσe−(x−µ)2/σ2, (30) Log-normal: pln(x; σ, µ) = 1 xpG(log(x)), (31) Lorentzian: pL(x; σ, µ) = σ 2π 1 (x −µ)2 + σ2 . (32) (30) (31) (32) Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 9 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 9 We will also consider an unconventional function that also has a ﬁnite bandwidth, but which is not log-convex σ We will also consider an unconventional function that also has a ﬁnite bandwidth, but which is not log-convex σ (33) pnc(x; σ) ∝e−x sin(2σx)2 cos(3σx)2. (33) We have chosen all these distributions because we can compute the functions p(m)(x) exactly for all sampling sizes, constructing the GR states for up to m = 14 qubits1, analyzing their structure and complexity. In Fig. 2a we plot the entanglement required to enlarge the quantum register by one bit, from m to m+1. This is the entropy S[ρ(m,1)] in the notation above. As shown in Fig. 2b, the bipartition never exceeds one e-bit of entanglement, and exhibits an exponential decay at large sizes that goes as 2−γm with γ between 1.73 and 1.84, depending on the simulation and probability distribution. The behavior is therefore more favorable than the bound from Eq. (12), which overestimates the entanglement. Fig. 2c shows that just like the entropy increase of adding one more qubit is small, the cumulative entropy obtained by studying all bipartitions of m + k qubits also remains small, and with a similar tendency. Moreover, for a detailed enough sampling with m = 14, there is no diﬀerence between the GR states and the simpler discretizations from Sect. 3.2, shown here with dash-dot lines. To test whether these favorable dependencies are artifacts of our choice of distributions, we have varied the parameters of the distributions and also tested situations where one of them (33) acquires more features. Fig. 1d shows the maximum entanglement entropy over all bipartitions as we change the parameter σ. The ﬁrst three probability distributions have a bounded entanglement below an e-bit. The non-log-convex distribution pnc(x) behaves slightly diﬀerent: increasing σ leads to the appearance of more peaks, that are more diﬃcult to describe. 1While m = 14 qubits do not challenge the computational capabilities of classical computers, in one dimension this precision leads to discretization error 6 × 10−6 so small that the plots do not change by enlarging the register. 3.5 Numerical study of 1D distributions This causes a steady increase in entropy, but this is slow enough that still facilitates an eﬃcient construct. 3.6 Extensions to more variables Challenging applications to ﬁelds such as Physics, ﬂuid dynamics or ﬁnance [26] require the study of states with many more, p(x1, x2 . . .). To better understand the scaling of entanglement and the complexity of the state, we have studied the discretization (9) of two- and three-dimensional Gaussian distributions p(⃗x) = 1 (2π)N/2det(Σ) exp −1 2⃗xT Σ−2⃗x , ⃗x ∈RN, (34) (34) ith covariance matrix Σ and zero mean, using the same number of qubits in all dimensions. As in the 1D problem, we will treat the quantum register as a one-dimensional arrange- ment of qubits, studying the entanglement over all 1D bipartitions. Naturally, the complex- ity of this discretization will depend on how we arrange the qubits. The simple straightfor- ward order (A) distributes the qubits sequentially, ﬁrst by coordinate, then by signiﬁcance. In this order, Gaussian states with a diagonal covariance matrix Σ = diag{σ1, σ2, . . .} be- come products states of one-dimensional distributions, such as those studied in Sects. 3.3 and 3.5. We also introduce the order (B), where qubits are ﬁrst sorted by signiﬁcance and only then by coordinate. This order is inspired by the renormalization group, and deeply similar to the multi-scale representation of 2D quantum image encodings [13]. For 10 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 2 1 0 1 2 x1/ max 2 1 0 1 2 x2/ max max min a) 0.000 0.005 0.010 0.015 0.020 0.025 0.030 0.2 0.4 0.6 0.8 1.0 min/ max 1.0 1.5 2.0 2.5 3.0 max(S) = /4 (B) = /4 (A) = 0 (A) b) exact, m = 14 MPS, m = 18 0 5 10 15 20 k 0.0 0.5 1.0 1.5 2.0 2.5 3.0 S[ (k, 2m k)]) exact (B) exact (A) c) 0 10 20 30 k 0.0 0.5 1.0 1.5 2.0 2.5 3.0 S[ (k, 2m k)]) MPS (B) MPS (A) c) Figure 3: (a) Two-mode Gaussian probability distribution with variances (σmin, σmax), rotated an angle θ. We work with a discretization interval [−7σmax, 7σmax]⊗2, compressing σmin down to 0.1, which is a 20dB squeezing of the variance. (b) Maximum entanglement of all bipartitions, for diﬀerent variances, orientations and orders. In solid lines, we plot the trivial sampling (9) with N ×m = 28 qubits in total. 3.6 Extensions to more variables We compare those plots with the same simulation using an MPS at a higher sampling (circles). We probe two angles θ = 0 and π/4 in Eq. (36), and two orders (35). (c) Entanglement entropies for all bipartitions of Nn = 24 qubits into (k, Nm −k), for the highly squeezed state σmin = 0.1σmax with θ = π/4. (d) Similar plot but for the MPS algorithm using a Nm = 36 qubits. 0.2 0.4 0.6 0.8 1.0 min/ max 1.0 1.5 2.0 2.5 3.0 max(S) = /4 (B) = /4 (A) = 0 (A) b) exact, m = 14 MPS, m = 18 2 1 0 1 2 x1/ max 2 1 0 1 2 x2/ max max min a) 0.000 0.005 0.010 0.015 0.020 0.025 0.030 0 5 10 15 20 k 0.0 0.5 1.0 1.5 2.0 2.5 3.0 S[ (k, 2m k)]) exact (B) exact (A) c) 0 10 20 30 k 0.0 0.5 1.0 1.5 2.0 2.5 3.0 S[ (k, 2m k)]) MPS (B) MPS (A) c) Figure 3: (a) Two-mode Gaussian probability distribution with variances (σmin, σmax), rotated an angle θ. We work with a discretization interval [−7σmax, 7σmax]⊗2, compressing σmin down to 0.1, which is a 20dB squeezing of the variance. (b) Maximum entanglement of all bipartitions, for diﬀerent variances, orientations and orders. In solid lines, we plot the trivial sampling (9) with N ×m = 28 qubits in total. We compare those plots with the same simulation using an MPS at a higher sampling (circles). We probe two angles θ = 0 and π/4 in Eq. (36), and two orders (35). (c) Entanglement entropies for all bipartitions of Nn = 24 qubits into (k, Nm −k), for the highly squeezed state σmin = 0.1σmax with θ = π/4. (d) Similar plot but for the MPS algorithm using a Nm = 36 qubits. a distribution with two random variables, x1 and x2, using three qubits per variable, the two orders read Order (A): \|s1s2⟩→\|s1 1⟩\|s2 1⟩\|s3 1⟩\|s1 2⟩\|s2 2⟩\|s3 2⟩, and (35) Order (B): \|s1s2⟩→\|s1 1⟩\|s1 2⟩\|s2 1⟩\|s2 2⟩\|s3 1⟩\|s3 2⟩. (35) Let us begin the discussion using a general two-dimensional covariance matrix Σ = O(θ) σmax 0 0 σmin ! OT , with O = cos(θ) sin(θ) −sin(θ) cos(θ) ! . 3.6 Extensions to more variables (36) (36) One quadrature is squeezed by a factor σmin/σmax, while rotating the frame of reference an angle θ, as shown in Fig. 3a. We sample this probability, reconstructing the exact wavefunction (9) with Nm = 28 qubits. As shown in Table 2, this size approaches the limits of a decent computer, using 2 gigabytes of data in real double precision form. In contrast, the same distributions using MPS, the techniques from Sect. 5.5 and up to Nm = 36 qubits consume less than 1 megabyte. One quadrature is squeezed by a factor σmin/σmax, while rotating the frame of reference an angle θ, as shown in Fig. 3a. We sample this probability, reconstructing the exact wavefunction (9) with Nm = 28 qubits. As shown in Table 2, this size approaches the limits of a decent computer, using 2 gigabytes of data in real double precision form. In contrast, the same distributions using MPS, the techniques from Sect. 5.5 and up to Nm = 36 qubits consume less than 1 megabyte. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 11 Discretization Nm σmin/σmax θ exact size order MPS size 16, 3842 28 1 0 268 × 106 (2 Gb) (A) 1,040 (8 kb) " 28 0.1 π/4 " (A) 126,628 (1 Mb) " 28 0.1 π/4 " (B) 9,388 (71 kb) 262, 1442 36 0.1 π/4 69 × 109 (524 Tb) (A) 183,220 (1 Mb) " 36 0.1 π/4 " (B) 10,626 (80 kb) 20483 33 0.1 π/4 8.6 × 109 (64 Gb) (A) 825,922 (6.6 Mb) " 33 0.1 π/4 " (B) 156,720 (1.22 Mb) Table 2: Summary of resources to describe numerically the Gaussian probability distributions with covariance matrices given by (36) and (37), using m qubits per dimension, either in an exact form— storing all values p(m)(xs)—or in the compact MPS representation (29). Table 2: Summary of resources to describe numerically the Gaussian probability distributions with covariance matrices given by (36) and (37), using m qubits per dimension, either in an exact form— storing all values p(m)(xs)—or in the compact MPS representation (29). Table 2: Summary of resources to describe numerically the Gaussian probability distributions with covariance matrices given by (36) and (37), using m qubits per dimension, either in an exact form— storing all values p(m)(xs)—or in the compact MPS representation (29). The output of these simulations is shown in Fig. 2. 3.6 Extensions to more variables When θ = 0, the wavefunction \|p(m) 2d ⟩is a product state \|p(m)(σmax)⟩⊗\|p(m)(σmin)⟩of two one-dimensional distributions. As shown in Fig. 2b (blue solid), the maximum entanglement over all one-dimensional bipartitions is less than one e-bit, consistent with Sect. 3.5. To grow the entanglement we must combine squeezing and rotation, recreating a two-mode squeezed state. From the theory of Gaussian states, the entanglement should be maximal for θ = π/4 and it should diverge with the squeezing. Our simulations conﬁrm this prediction for the (A) order. Fig. 3b (orange, solid) shows that the maximum bipartite entanglement between our qubit variables grows as (σmax/σmin)1/4. This entanglement is also spread all along the chain of qubits, as seen in Fig. 3c. We can even improve on these results. As shown in Fig. 3b, if we adopt the renormalization order (B), all states can be described with N = 2 e-bits of entanglement. Moreover, the entanglement distribution concentrates around the most signiﬁcant qubits [cf. Fig. 3c (dashed, green)], producing signiﬁcantly smaller tensors. Despite the growth of entanglement, the quantum states that we create always admit a compact MPS representation that beats the classical approach of storing the full wave- function. As shown in Table 2, a highly-correlated discretization in the (A) order with 14 and 18 bits per coordinate, requires 1Mb of ﬂoating point real numbers in MPS form. The same states stored using the (B) order, take, in the worst scenario σmin = 0.1σmax with θ = π/4, just about 80 kilobytes of information. All this is to be compared to the 2 Gb and 524 Tb of data required to write down the wavefunctions of 28 and 36 qubits. 0.2 0.4 0.6 0.8 1.0 1.0 1.5 2.0 2.5 3.0 = /4 (B) = /4 (A) = 0 (A) a) b) Figure 4: (a) Maximum entanglement over all bipartitions for a three-dimensional Gaussian state (37), computed with exact wavefunctions (m = 7 bits per mode, lines) or MPS (m = 9 bits per mode, circles), using diﬀerent squeezing rates, angles and orders. (b) Illustration of the three-dimensional Gaussian probability distribution for σmin = 0.5σmax. Figure 4: (a) Maximum entanglement over all bipartitions for a three-dimensional Gaussian state (37), computed with exact wavefunctions (m = 7 bits per mode, lines) or MPS (m = 9 bits per mode, circles), using diﬀerent squeezing rates, angles and orders. 3.6 Extensions to more variables (b) Illustration of the three-dimensional Gaussian probability distribution for σmin = 0.5σmax. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 12 We have performed the same study using three-dimensional Gaussian states. For con- creteness, we have focused on a three-mode squeezed state that starts with a diagonal matrix diag(σmin, σmax, σmin) and performs two identical rotations around the X and Z axes, with angles θx = θy = θ Σ = Ox(θx)Oz(θy) σmax 0 0 σmin ! Oz(θz)T Ox(θx), with (37) Ox =    1 0 0 0 cos(θ) sin(θ) 0 −sin(θ) cos(θ)   , Oz =    cos(θ) sin(θ) 0 −sin(θ) cos(θ) 0 0 0 1   . (37) We show results for a large, exact simulation with Nm = 21 qubits, which amounts to 2563 points and 128 Mb of data, together with an MPS that is directly built with 33 qubits, a sampling of 20483 points. As in the two-dimensional case, the combination of squeezing and rotation deforms the state, which as shown in Fig. 4b becomes a tilted rugby ball. Once more, the unrotated state remains weakly entangled. It is a product of three one- dimensional probabilities, and the entanglement never exceeds one 1 e-bit [cf. Fig. 4a (blue, solid)]. The squeezing and rotation leads to a divergence of the maximum entanglement, but this divergence is once more cured by the (B) order. This importance-based structure brings down the entanglement to about 3 e-bits and a reduction of 54,000 in the information required. We conjecture that for these Gaussian states—and other smooth functions—the (B) order consumes at most N e-bits, giving a scaling of resources O(22N ×N) in both the time and memory costs of reproducing the probability distribution. 4 MPS simulation of quantum registers We have seen that it is possible to encode single and multimode probability distributions (and general smooth functions!) in quantum registers, that these states are typically weakly entangled and admit an eﬃcient MPS representation. In the remaining of this work we take this idea a step further, advocating the use of MPS to study multivariate analysis problems, by encoding the solutions to those problems in the state of a ﬁctitious quantum register, and representing and manipulating this register using MPS-simulation techniques. In order to explain how those algorithms work, we need to review the representation of wavefunctions and operators, and the basic ingredients in the MPS toolbox—tensor con- traction and reordering, tensor renormalization, time evolution, etc—that will be used in Sect. 5 to implement actual algorithms. More precisely, Sect. 4.1 introduces two represen- tations of multivariate functions: one following the precepts from Sect. 3.2, and another one that improves the computation of expected values (Sect. 4.3) and equation solving. We will also discuss the algorithm of MPS simpliﬁcation (Sect. 4.4), which is an essential tool to implement all numerical analysis approximation schemes. We also refer the reader to a more practical introduction [20] and to our SeeMPS library [27, 28] for details on how these methods are implemented in practice. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 4.1 Quadratic and linear representation Our overarching goal for the rest of this work is to encode multivariate functions using a virtual quantum register, and to store this register eﬃciently as an MPS. If the function we want to encode is non-negative—such as a probability distribution—, we can follow Sect. 3, identify √p with a quantum register wavefunction (2) and use the MPS representation (29) 13 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. Figure 5: Quadratic MPS representation, where functions are mapped to the amplitude of a wavefunc- tion (as MPS) and observables are mapped to operators (as MPO). Expected values ¯f = ⟨p\| ˆOf\|p⟩are obtained by contracting three layers of tensors with physical dimensions d and bond dimensions ∼χ. This has a cost O(3d2χ4) and causes each tensor to appear twice—in other words, ¯f is a quadratic function w.r.t. each tensor. Figure 5: Quadratic MPS representation, where functions are mapped to the amplitude of a wavefunc- tion (as MPS) and observables are mapped to operators (as MPO). Expected values ¯f = ⟨p\| ˆOf\|p⟩are obtained by contracting three layers of tensors with physical dimensions d and bond dimensions ∼χ. This has a cost O(3d2χ4) and causes each tensor to appear twice—in other words, ¯f is a quadratic function w.r.t. each tensor. to compress it. In this representation, observables f(x) become Matrix-Product operators (MPO’s [29]) ˆO(m) f = X α,s Bs1,r1 β1 Bs2,r2 β2,β3 · · · Bsm,rm βm \|s1, s2 . . . sm⟩⟨r1, r2 . . . rm\| . (38) (38) We call this the quadratic MPS representation because the mean value of an observable ¯f ∼⟨p\| ˆOf\|p⟩is a quadratic function of any of the tensors in the MPS state [cf. Fig. 5]. f ⟨p\| f\|p⟩ q y [ g ] In the alternative encoding from equation (3), both observables f(x) and probability distributions p(x) become unnormalized vectors in a Hilbert space In the alternative encoding from equation (3), both observables f(x) and probabilit distributions p(x) become unnormalized vectors in a Hilbert space \|f(m)⟩= X s f(m)(s) \|s⟩= X α,s F s1 β1 F s2 β2,β3 · · · F sm βm \|s1, s2 . . . sm⟩, (39) \|p(m)⟩= X s p(m)(s) ⟨s\| = X α,s As1 α1As2 α1,α2 · · · Asm αm \|s1, s2, . . . , sm⟩. 4.1 Quadratic and linear representation (40) (39) (40) In this representation, each probability state is a linear form that maps observables to expected values, or vice versa ¯f = X s f(s)p(s) = ⟨p\|f⟩≃ Z f(x)p(x)dx. (41) (41) We will call this strategy the linear MPS representation because ¯f is a linear function with respect to any of the tensors in \|p⟩or \|f⟩. We will call this strategy the linear MPS representation because ¯f is a linear function with respect to any of the tensors in \|p⟩or \|f⟩. 4.2 Memory cost Let us denote χf, χp or simply χ the largest bond dimensions to encode those functions as MPS. Assuming an N-dimensional volume, discretized with m qubits per dimension, the space required by an MPS and an MPO scales as O(Nmχ2). Since χ dominates this scaling, we need to understand how this dimension behaves in typical problems. The answer to this question is connected to the bipartite entanglement that is stored in the MPS representations. Since MPS’s are obtained through a recursive Schmidt decompo- sition [30], the entanglement entropy over any sequential bipartition of the state is bounded by S ≤log2(χ). Conversely, if the maximum entanglement is S, we will expect that the 14 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. a) b) Figure 6: Linear MPS representation. (a) Both observables \|f⟩and a probability distributions \|p⟩ have MPS representations with bond dimensions of order χ, and they combine to give a mean value ¯f = ⟨p\|f⟩. (b) When there are m qubits, the optimal contraction is sequence of m × 2 steps, with the structure shown here, taking O(2dχ3) operations. a) b) b) Figure 6: Linear MPS representation. (a) Both observables \|f⟩and a probability distributions \|p⟩ have MPS representations with bond dimensions of order χ, and they combine to give a mean value ¯f = ⟨p\|f⟩. (b) When there are m qubits, the optimal contraction is sequence of m × 2 steps, with the structure shown here, taking O(2dχ3) operations. bond dimension scales as χ ∼2S. Thus, for the smooth distributions from Sect. 3.5, where S ∼N, we expect a scaling of resources of the form2 memory ∼O(mN22S+1) ∼O(mN22N+1) ∼O N22N+1 log2(1/εint) (42) (42) Note that 2m ∼εint grows with the desired discretization precision. Therefore, if we consider this quantity as ruling parameter, the asymptotic tendency 22N+1 log2(1/εint) represents an exponential saving over the space O(2Nm) ∼O(1/εN int) required to store the full wavefunction in general classical algorithms. Note that 2m ∼εint grows with the desired discretization precision. Therefore, if we consider this quantity as ruling parameter, the asymptotic tendency 22N+1 log2(1/εint) represents an exponential saving over the space O(2Nm) ∼O(1/εN int) required to store the full wavefunction in general classical algorithms. This exponential improvement is similar in origin to the one in quantum computers, as it also exploits the rapid growth of the Hilbert space with the number of qubits. 2This type of cost analysis is standard in the MPS and tensor network literature. You may consult Ref. [20] for some general analysis of MPS and MPO contraction and simpliﬁcation costs. 4.2 Memory cost We must remark that in the MPS representation the gain is heuristic: it only appears for distributions with nice sampling properties. There are inﬁnitely many problems where this improvement vanishes, due to the growth of S and the exponential blowup of the bond dimensions χ. However, it also seems that there are still many problems of interest where the MPS representation is useful, as we see below. 4.4 Approximating states In working with the MPS representation, we will frequently need to apply operators that distort and enlarge the tensors. This is corrected by a process known as MPS simpliﬁcation, which seeks the closest matrix-product state with the smallest bond dimensions, within a prescribed time and error tolerance. The simpliﬁcation is an optimization typically deﬁned with respect to the 2-norm distance between states argminφ in MPS∥φ −p∥2 = argminφd(φ, p). (43) (43) Here p is the state we wish to approximate and φ is the new MPS. The distance d(φ, p) = ⟨p\|p⟩+ ⟨φ\|φ⟩−2Re ⟨φ\|p⟩is a quadratic form with respect to the tensors in φ, which is optimized iteratively, sweeping across the MPS [20, 21, 31] in a two-site DMRG-like process. Here p is the state we wish to approximate and φ is the new MPS. The distance d(φ, p) = ⟨p\|p⟩+ ⟨φ\|φ⟩−2Re ⟨φ\|p⟩is a quadratic form with respect to the tensors in φ, which is optimized iteratively, sweeping across the MPS [20, 21, 31] in a two-site DMRG-like process. The cost of this optimization has two parts. The estimation of the linear form ⟨φ\|p⟩ involves a contractions like the ones shown in Fig. 6b, involving O(dχ3) operations per site. On top of this, we apply a two-site simpliﬁcation algorithm that optimizes pairs of tensors simultaneously, dynamically adapting the bond dimension. This has an extra cost O(4(dχ)3) due to the singular value decomposition. Thus, assuming that we need Tsweeps for convergence, the simpliﬁcation time cost grows as O(Tsweeps4d3χ3). In practical examples we ﬁnd that Tsweeps is very small: one or two sweeps reach the numerical precision of the computer, giving eﬃcient results. Finally, note that we can use the algorithm to construct a new state φ that approximates a linear combination of MPS argminφ in MPS∥φ − k X i=1 αipi∥2. (44) (44) This has a linear increase in the cost, O(kTsweeps4d3χ3), that has been exploited in other algorithms such as time evolution [21]. This has a linear increase in the cost, O(kTsweeps4d3χ3), that has been exploited in other algorithms such as time evolution [21]. 4.3 Integrals and expected values In the linear MPS representation, the scalar product ⟨f\|p⟩is an approximation to the integral between both functions R f(x)p(x)dx. As sketched in Fig. 6a, there is an optimal contraction scheme that starts from one boundary—the left-most site in Fig. 6a—, and sequentially contracts with one tensor from \|p⟩and one from \|f⟩. The optimal sequence of contractions in Fig. 6b demands O(dχ3) operations in 2Nm steps, with an estimated cost O(2dNmχ3). We can perform a similar analysis for the quadratic MPS representation, where mean values ¯f = ⟨p\| ˆOf\|p⟩involve contracting the state \|p⟩twice with the MPO ˆOf Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 15 that represents the observable. In this case the optimal procedure is slightly more involved, with a higher asymptotic cost O(Nm × 4χ4). In order to compare these algorithms with other classical methods, we have to introduce the accuracy of the estimate. In the MPS representation the only error we make is the discretization of N variables with m qubits, which scales as εint ∼N2m. Thus, the MPS approximation to the integral demands a time Tconstr + O(N log2(1/εint/N) × 2χ3), where Tconstr is the time to build the MPS and the rest is a logarithmically growing cost associated to the contraction. We can compare this with Monte Carlo sampling [18], a good and general method for integration. The errors in this technique arise from the statistical uncertainty εsample ∼1/ √ M. This error decays slowly with the number of iterations M, giving a time cost O(1/ε2). The MPS therefore has the potential of providing an exponential speedup, given that (i) the bond dimension χ remains small and (ii) the cost of constructing the MPS states is also bounded. 4.5 Multiplication by functions and operators In many algorithms below we will need to construct a state \|fp⟩that approximates the product of two sampled functions f(x)p(x). This operation can be implemented eﬃciently in at least four cases, which we now discuss. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 16 Given an arbitrary complex constant c and a discretization of N variables with m qubits per dimension, there exist MPO’s for f(x) = cx, cx2 and exp(cx) using bond dimensions 2, Nm and 1, respectively. As illustration, let us discuss the implementation of cx. This operator is an MPO (38) with a bond dimension of size χ = 2 that keeps track of whether any operator has been applied. The n −th tensor reads Bs′ nsn a,b = δs′n,sn          cL2−nsn, a = 0, b = 1 1, a = b = 1, n > 1 1, a = b = 0, 0, else. (45) (45) More generally, we can write the exponential of any QUBO formula as a product of MPO’s with bond dimension 2 exp   K X i,j=1 Qijsisj  = K Y k=1 exp  si X j Qijsj  . (46) (46) The MPO’s inside the product are constructed with simple tensors. In particular, for the k−th step, the tensors read The MPO’s inside the product are constructed with simple tensors. In particular, for the k−th step, the tensors read Bs′ nsn a,b = δs′nsn      δa0δb0, n < k, δbs1δa0 exp(Q11sn), n = k, δab exp(2aQansn), k < n. (47) (47) Using this idea, we can decompose any exponential or Gaussian, such as exp(cx2 + dx) or exp(P nm cnmxnxm), using Nm layers of MPO’s. This technique is discussed in Sect. 5.5 when we construct MPS. For other formulas, we ﬁnd alternative approaches. One is to ﬁnd an MPO represen- tation of the operator ˆOf. This is expected to be a simple tensor contraction for smooth functions, but the cost of computing it in general may be signiﬁcant, unless we combine it with strategies such as interpolation (cf. Sect. 5.2.2). Another alternative is to approxi- mate f(x) in Taylor series and use multiplications by constants and by x, combined with simpliﬁcation stages, to approximate the product. A ﬁnal approach is to use algorithms from quantum computing to implement the equivalent transformations [32, 33]. 4.5 Multiplication by functions and operators The in- teresting thing in this approach is that, since we are working with artiﬁcial registers, we can forego many of the auxiliary qubits, imposing success in all operations. 5 Multivariate analysis on the MPS-simulated quantum register After this brief review of MPS method, we will now introduce speciﬁc, higher level al- gorithms for interpolation, Fourier analysis, diﬀerentiation, solving PDE’s and creating MPS for a sampled function. Unless otherwise stated, all algorithms are implemented us- ing the linear MPS encoding from equations (3) and (40). Whenever possible we provide also a comparison with the asymptotic performance of state-of-the-art classical algorithms, discussing issues of eﬃciency, accuracy and numerical stability. 5.1 Fourier transforms and frequency analysis Assume a function p(xs) uniformly sampled over 2m points in an interval xs ∈[a, b] of size L = \|b −a\|. We can deﬁne 2m distinct Fourier modes ˜ψr(xs) ∝exp(iqrxs) for qr = 2πr/L Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 17 0 5 10 15 j (step) 0.00 0.25 0.50 0.75 1.00 max S[ (k, m k)] d) j 1 U2c 5 0 5 xs 0.00000 0.00005 0.00010 0.00015 p(xs) a) 0.0 0.2 0.4 0.6 0.8 1.0 s/2m 0.0 0.1 0.2 0.3 [ p](s) b) 0 5 10 k 0.00 0.25 0.50 0.75 1.00 S[ (k, m k)] c) U2c Figure 7: (a) Discretized Gaussian probability distribution with σ = 1, µ = 0, over 15 qubits, and (b) quantum Fourier transform. (b) Entanglement entropy with respect to all bipartitions of a probability distribution ψ (blue, circles), its quantum Fourier transform Fψ (orange), and the two’s complement of this (green, squares). (c) Maximum entanglement entropy for all bipartitions for the original state (circle), diﬀerent stages of the QFT (orange) and the QFT after a two’s complement transformation. 5 0 5 xs 0.00000 0.00005 0.00010 0.00015 p(xs) a) 0 5 10 k 0.00 0.25 0.50 0.75 1.00 S[ (k, m k)] c) U2c c) 0 5 10 15 j (step) 0.00 0.25 0.50 0.75 1.00 max S[ (k, m k)] d) j 1 U2c Figure 7: (a) Discretized Gaussian probability distribution with σ = 1, µ = 0, over 15 qubits, and (b) quantum Fourier transform. (b) Entanglement entropy with respect to all bipartitions of a probability distribution ψ (blue, circles), its quantum Fourier transform Fψ (orange), and the two’s complement of this (green, squares). (c) Maximum entanglement entropy for all bipartitions for the original state (circle), diﬀerent stages of the QFT (orange) and the QFT after a two’s complement transformation. and r = 0, 1, . . . 2m. We can also deﬁne a Fourier transform and r = 0, 1, . . . 2m. We can also deﬁne a Fourier transform ˜p(qr) = [Fp](qr) = 1 2m/2 2m−1 X s=0 exp (−iqr(xs −a)) p(xs) (48) = 1 2m/2 2m−1 X s=0 exp (−i2πrs/2m) p(xs), (48) which returns the expansion of the original function in the Fourier basis. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.1 Fourier transforms and frequency analysis Fortunately, there exists a very eﬃcient classical algorithm, known as the Fast Fourier Transform (FFT), which implements this formula with a cost that is almost proportional to the size of our discretrization [18], that is O(m2m) for one dimension or O(Nm2Nm) for an N-dimensional function. The Quantum Fourier Transform (QFT) is the extension of the classical operator F to the quantum realm. The QFT operator ˆF takes a quantum state with wavefunction p(s) and converts it into its Fourier transform ˆF \|p⟩= X s p(s) ˆF \|s⟩= X r ˜p(r) \|r⟩. (49) (49) The QFT is deﬁned by the action on the quantum register states The QFT is deﬁned by the action on the quantum register states ˆF \|s⟩= 1 2m/2 X s ei2πss′/2m \|s′⟩. (50) (50) This operation can be implemented using m+1 layers of quantum gates, as F = UﬂipFm · · · F1. The last operation, Uﬂip just reverses the order of the qubits in the quantum register. The Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 18 i-th layer Fi contains a Hadamard gate Hi on the i-th qubit, followed by m −i controlled rotations with respect to the following m −i qubits i-th layer Fi contains a Hadamard gate Hi on the i-th qubit, followed by m −i controlled rotations with respect to the following m −i qubits Fi = m Y j=i+1 exp i 2π 2j−i σz i ⊗σz j exp iπ 2 σy i . (51) (51) Each of the Fi is an MPO with bond dimension χ = 2 and tensors Bs′ n,sn 0,0 = δs′n,sn, n < i, (52) B s′ isi 0,s′ i = Hs′ i,si, i-th qubit, B s′ j,sj si,si = exp i 2π 2j−i sjsi δs′ jsj, j > i. (52) Applying the QFT on an MPS requires contracting the Nm operators Fi and simplify- ing the resulting states. The cost of each simpliﬁcation stage SimpNm dominates the per- formance of the algorithm, so that we can assume a scaling O(Nm×SimpNm). The cost of one simpliﬁcation stage SimpNm ∼O(TsweepsNm4d3χ3) is very much problem dependent, as it depends on the number of required sweeps and entanglement. For smooth functions we expect that the entanglement will be bounded, both because of the estimates above, and because smooth functions will also tend to be concentrated in Fourier space. 5.2.1 Taylor expansions Let us consider a scenario in which we have discretized the interval [a, b] uniformly with 2m points, and we want to add 2m extra points, moving to m + 1 qubits. Let us call xs the original variables and x′ r the new sampling, which satisﬁes (55) x′ 2s = x2s, x′ 2s+1 = xs + δm/2. (55) We assume that the values at xs determine those at x′ 2s p(m+1)(x′ 2s) = p(m)(xs), s = 0, 1, . . . 2m −1, (56) (56) and we only need to extrapolate the values at the odd sites x′ 2s+1. In order to do so, we can assume that our function is analytic and admits a Taylor expansion to some ﬁnite order, which gives the following approximation p(xs + δm+1) = 1 2 [p(xs) + p(xs+1)] + O(δ2 m+1). (57) (57) Note that the cost of this classical interpolation algorithm is proportional to the size of the destination vector, thus O(2N(m+1)) ∼O(ε−N int ), considering the precision of the ﬁnal discretization εint ∝2−(m+1). We can translate equations (56) and (57) into an algorithm that extends a sampling with m qubits into another one with m + 1. In a way that resembles very much equation (7) , but which is deﬁnitely not unitary, we add one least signiﬁcant qubit at the end \|pm+1⟩= \|pm⟩\|0⟩+ 1 2( ˆS−+ 1) \|pm⟩\|1⟩. (58) (58) The ladder operators ˆS± increase or decrease the quantum register by one, displacing the function we encoded The ladder operators ˆS± increase or decrease the quantum register by one, displacing the function we encoded ˆS+ \|s⟩= ( \|s + 1⟩, s < 2m 0, else ˆS−= ( ˆS+)†. (59) (59) Instead of using reversible operations (Toﬀoli, CNOT), we implement the MPO as a smaller, irreversible and classical circuit with one carry bit that propagates through the bond-dimension. This requires a single tensor C ˆS+ = X C s′ 1,s1 0,a1 C s′ 2,s2 a1,a2 · · · Cs′ m,sm am,1 \|s′ 1, s′ 2 . . . s′ m⟩⟨s1, s2 . . . sm\| . (60) (60) The tensor Cs′,s a,b is nonzero only for s′ = s ⊕b, and a = s ∧b. The MPO ˆS−is obtained by simply exchanging the indices as ˆS−= X C s′ 1,s1 0,a1 C s′ 2,s2 a1,a2 · · · Cs′ m,sm am,1 \|s1, s2 . . . 5.2 Interpolation 5.2 Interpolation Interpolating means approximating values of a discretized function on points that were not initially considered. We discuss two techniques for interpolating from a quantum register with m qubits to a new register and discretization with m + k qubits. The ﬁrst method uses ﬁnite diﬀerences to extrapolate new points as linear combinations of previous values. The second method is a spectral technique based on Fourier transformations that, as we will show, can be exponentially more accurate for ﬁnite bandwidth functions. Interpolating means approximating values of a discretized function on points that were not initially considered. We discuss two techniques for interpolating from a quantum register with m qubits to a new register and discretization with m + k qubits. The ﬁrst method uses ﬁnite diﬀerences to extrapolate new points as linear combinations of previous values. The second method is a spectral technique based on Fourier transformations that, as we will show, can be exponentially more accurate for ﬁnite bandwidth functions. 5.1 Fourier transforms and frequency analysis In that case we can link the cost of one simpliﬁcation stage SimpNm ∼O(log2(1/εint) and of the full QFT O(log2(1/εint)2)) to the precision of the discretization εint ∝2−m. This is exponen- tially faster than the classical FFT O(Nm2Nm) ∼O(log2(1/εint)/εN int). It is interesting to note that other authors have considered using the QFT in a classical context [34, 35], but they never observed a real speed-up—more like a 20-fold slow down—because of working with the complete wavefunction and not with the MPS-simulated quantum register. As example, take the Gaussian probability distribution from Fig. 7a. Its QFT is a highly concentrated state, another Gaussian in momentum space shown in Fig. 7b. We know that the entanglement entropy of the transformed state is upper-bounded by 1 e-bit (see Sect. 3.5). This is what we see not only for the ﬁnal state—orange line in Fig. 7c—, but also when we analyze all stages of the transform F1ψ, F2F1ψ, etc [cf. Fig. 7d.] Let us inspect more carefully the Fourier transformed wavefunction Fig. 7b. Note how the wavefunction concentrates on both sides of the interval, s ≃0 and s ∼2m −1. This is caused by the mapping from the non-negative quantum register states s, to the actual momenta, and which has the representation ˆk = X s k¯s \|s⟩⟨s\| , with ks = 2π L ¯s ∈ π L, π L . (53) (53) Here ¯s is the two’s complement of the binary number s = s1s2 . . . sm ¯s = (1 −s1) m X n=2 2m−nsn −s1 " 1 + m X n=2 2m−nsn # . (54) (54) It is useful to implement a two’s complement operation that ﬂips all qubits conditioned on the state of the sign qubit, U2c = \|s1, s2, . . . , sm⟩→\|s1, s1 ⊕s2, . . . , s1 ⊕sm⟩. When we apply this operator to the Fourier transformed state, U2cFψ, we ﬁnd that the amount of entanglement surprisingly drops down, almost close to zero for all bipartitions—see Figs. 7c-d. This hints at the fact that the signed quantum register is a much better variable for describing this (and probably other) symmetric probability distributions. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 19 5.2.1 Taylor expansions sm⟩⟨s′ 1, s′ 2 . . . s′ m\| , (61) (61) Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 20 5 0 5 xs 0.0 0.1 0.2 p(5)(xs) a) 5 0 5 ks 0.0 0.1 0.2 [ p](s) c) 5 0 5 xs 0.0 0.1 0.2 p(5)(xs) a) 5 0 5 xs 0.0000 0.0025 0.0050 0.0075 p(10)(xs) b) 5 0 5 ks 0.0 0.1 0.2 [ p](s) c) 200 0 200 ks 0.0 0.1 0.2 [ p](s) d) Figure 8: (a) Gaussian probability distribution sampled with m = 5 qubits and 32 points. (b) Fourier interpolation and exact sampling with m = 10 qubits and 1024 points. Both samplings are indistin- guishable. (c) Fourier spectrum of the original distribution. Note how higher frequency components are negligible. (d) Fourier transform of the interpolated function. We have added zero values over 25 sites at higher momenta, outside the axes. Figure 8: (a) Gaussian probability distribution sampled with m = 5 qubits and 32 points. (b) Fourier interpolation and exact sampling with m = 10 qubits and 1024 points. Both samplings are indistin- guishable. (c) Fourier spectrum of the original distribution. Note how higher frequency components are negligible. (d) Fourier transform of the interpolated function. We have added zero values over 25 sites at higher momenta, outside the axes. and the linear combination algorithm (58) can be implemented using the simpliﬁcation techniques from Sect. 4.4 Heuristically, the MPS algorithm can be exponentially more eﬃcient than trivial inter- polation. Since the MPO’s have very small bond dimensions, they can be integrated in the simpliﬁcation phase [20], which dominates the overall cost [cf. Table 1]. More precisely, in the case of smooth functions with low-entanglement representations, the MPS tensors have a small bond dimension and the simpliﬁcation will require SimpN(m+1) ∼O(log2(1/εint)) operations, which behaves heuristically better than the scaling of the classical algorithm. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.3 Diﬀerentiation In numerical analysis, there are two main ways to estimate the derivative of a discretized function. The ﬁrst method is called ﬁnite diﬀerences, because it relies on linear combina- tions of the function p(xs) and its displacements p(xs ± nδx). The second type of methods are called spectral methods, because they work with the Fourier expansion from (62). Both methods have simple translations to the language of MPO’s and MPS’s. 5.2.2 Fourier interpolation We start with a Fourier transform and two’s complement over m qubits. We then insert k qubits in positions 2 to k + 1. These are bits that encode very high frequencies and which are populated with zeros, as we do not need any ﬁner details in the sampled function. We ﬁnally take the enlarged register and invert both the two’s complement and the Fourier transform. As illustrated in Fig. 8 this is a powerful technique that can reconstruct a Gaussian using 210 = 1024 points out of a discretized Gaussian with 25 = 128 points, with negligible error. Note that the classical and MPS-simulated version of Fourier interpolation contain the same ingredients: zero-padding in between two discrete Fourier transforms, which dominate the scaling of the algorithms. However, when we operate with MPS-encoded functions, the improved performance of the QFT with MPS can provide an exponential speedup in practical situations. Following Section 5.1, if we extend the discretization, interpolating from a grid with 2m points to one with 2m+k, the classical algorithm based on the FFT will scale as O(N(m + k)2N(m+k)) ∼O(log2(1/εint)/εN int) if we consider the discretization error of the ﬁnal lattice εint. The MPS on the other hand will scale as O(N(m + k) × SimpN(m+k)). If the practical bond dimension remains bounded χ, this implies a behavior O(TsweepsN2(m + k)24d3χ3) ∼O(log2(1/εint)2N2χ3). This is exponentially faster in many useful situations. 5.2.2 Fourier interpolation The accuracy of linear interpolation is constrained by the initial sampling, O(δ2 m). This seems to contradict the Nyquist-Shannon theorem, according to which a function with a bounded spectrum only needs to be sampled with a frequency of 2 × νmax for a perfect interpolation. Take for instance a Gaussian probability distribution (30) with width σ, which has been sampled in the interval [−8σ, 8σ]. Its Fourier transform is a normal distri- bution with center ν = 0 and width 1/σ. We can say that the information beyond ν = 4/σ, is exponentially suppressed. According to Nyquist’s theorem, we can exactly reconstruct a Gaussian function by sampling it with period σ/8. For the conditions above, that means 8/σ ×16σ ∼128 points stored in 7 qubits. However, if we attempt linear interpolation, we typically will make a bounded error that is ﬁxed by the initial sampling, O(δx ∼1/16). A well known solution is to do the interpolation in frequency space. When we perform a discrete Fourier transform, we are decomposing the sampled function p(xs) as a sum of discrete Fourier modes ψ(k¯s′) ∝exp(ik¯s′xs). We can use this to reconstruct a continuous approximation to the original function p(x) in what is known as Fourier interpolation [36] p(x) ≃ 1 p \|b −a\| 2m−1 X s=0 exp(ik¯sx) h Fp(m)i (k¯s), x ∈[a, b]. (62) (62) 21 This continuous approximation can then be resampled with as ﬁne a grid as needed. The technique is so ubiquitous that it is available in standard numerical toolboxes, such as Matlab. We can implement the approximation and re-sampling very eﬃciently. If we wish t enlarge the number of qubits from m to m + k, we compute \|p(m+k)⟩= ˆF(m+k)Um+k 2c h U2c ˆF(m) \|p(m)⟩ ⊗\|02, 03, . . . , 0k+1⟩ i . (63) (63) We start with a Fourier transform and two’s complement over m qubits. We then insert k qubits in positions 2 to k + 1. These are bits that encode very high frequencies and which are populated with zeros, as we do not need any ﬁner details in the sampled function. We ﬁnally take the enlarged register and invert both the two’s complement and the Fourier transform. As illustrated in Fig. 8 this is a powerful technique that can reconstruct a Gaussian using 210 = 1024 points out of a discretized Gaussian with 25 = 128 points, with negligible error. 5.3.2 Fourier approximations to derivatives Since Fourier interpolation works so well, we can use it to approximate the action deriva- tives at arbitrarily high orders. Take any diﬀerential operator G(∂x), such as ∂x, ∂2 x, a∂x + b∂2 x, etc. The action of this operator on the Fourier interpolated continuous function (62) is very simple G(∂x)p(x) ≃ 1 p \|b −a\| 2m−1 X s=0 G(ik¯s) exp(ik¯sx) h Fp(m)i (k¯s), x ∈[a, b]. (67) (67) This formula tells us to Fourier transform the discretized function, multiply it by some factors G(iks) and then Fourier-transform back to position space. In a classical scenario, where we store a function using 2Nm values, the cost of this algorithm is dominated by the FFT O(Nm2Nm) ∼O(N log2(1/εint)/εN int). This formula tells us to Fourier transform the discretized function, multiply it by some factors G(iks) and then Fourier-transform back to position space. In a classical scenario, where we store a function using 2Nm values, the cost of this algorithm is dominated by the FFT O(Nm2Nm) ∼O(N log2(1/εint)/εN int). We can translate this recipe to our representation of functions in a quantum regis- ter. This leads to a non-unitary transformation, with two QFT’s and multiplication by a function of the momentum operator (53) G(∂x) \|p(m)⟩= ˆF−1G(iˆk) ˆF \|p(m)⟩. (68) (68) Even though this is a non-unitary operation, it can be computed using the MPS representa- tion. In this case the algorithmic cost is equivalent to that of a QFT, plus the product with the operator G(iˆk). Fortunately, for ﬁrst and second order derivatives G(iˆk) ∼aˆk + bˆk2 has an eﬃcient MPO representation and we revert back to the good heuristic scaling of SimpNm ∼O(Nχ3 log2(1/εint)) for smooth functions. 5.3.1 Finite diﬀerences We illustrate this technique using standard ﬁnite-diﬀerence approximations to ﬁrst and second spatial derivatives [18], ∂ ∂xp(x) ≃p(x + δx) −p(x −δx) 2δx + O(δx2), (64) ∂2 ∂x2 p(x) ≃p(x + δx)p(x −δx) −2p(x) δx2 + O(δx). (65) (64) (65) Note how the error of the approximation depends on the discretization step δx ∼(b−a)/2m of the uniform sampling (1). In classical algorithms, these formulas are applied onto a vector of values representing the function discretized over an N-dimensional grid with 2m points along each dimension. In this scenario, the cost of this operation is proportional to the size of the vector that encodes the function O(2Nm) ∼O(ε−N int ). Note how the error of the approximation depends on the discretization step δx ∼(b−a)/2m of the uniform sampling (1). In classical algorithms, these formulas are applied onto a vector of values representing the function discretized over an N-dimensional grid with 2m points along each dimension. In this scenario, the cost of this operation is proportional to the size of the vector that encodes the function O(2Nm) ∼O(ε−N int ). ( ) ( int ) We can improve on this by using the MPS representation, which requires less memory to store the same function. Here, small changes or displacements on the grid xs ± δx Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 22 map to increments and decrements of the quantum register variable s. These motions a implemented using the ladder operators (59). Our ﬁnite-diﬀerence formulas become ∂x \|p⟩≃ ˆS+ −ˆS− 2δx \|p⟩, ∂2 xx \|p⟩≃ ˆS+ + ˆS−−1 δx2 \|p⟩. (66) (66) These formulas can be implemented as MPO’s of bond dimension 3, which can be eﬃciently contracted and simpliﬁed using the algorithm from Sect. 4.4. Unsurprisingly, as in the linear interpolation algorithm, the cost is dominated by the simpliﬁcation phase SimpNm ∼ O(log2(1/εint)Nχ3), which can be exponentially faster for functions that allow a low- entanglement representation with small bond dimension χ. Note that higher order approximations are also possible using the same operator or powers of it. Roughly, the bond dimension of the MPO that implements a ﬁnite diﬀerence formula grows linearly with the order of the approximation, just like the number of non- diagonals in its matrix representation. This is a moderate cost that makes diﬀerentiation an approachable routine in higher level algorithms. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.4 Solving partial diﬀerential equations One of the main applications of all these techniques is the study of how multivariate functions evolve in time, when subject to one of many partial diﬀerential equations. We focus our discussion on the Fokker-Planck equation in one dimension ∂ ∂tp(x, t) = −∂ ∂x[µ(x, t)p(x, t)] + ∂2 ∂x2 [D(x, t)p(x, t)], (69) (69) 23 0 2 4 6 8 10 t 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 b) x = x 2 = (x x)2 x, theory 2, theory 10 5 0 5 10 0 2 4 6 8 10 a) 0.000 0.001 0.002 0.003 0.004 0.005 0.006 0.007 Figure 9: Solution of the Fokker-Planck equation using MPS and ﬁnite diﬀerences. (a) Density plot of the probability distribution evolved with the Fokker-Planck equation (69) and algorithm (73). In dashed lines we plot the expected value ¯x(t) and the curves ¯x(t) ± σ(t). (b) Center ¯x(t) and variance σ2(t) of the a probability distribution. We use D = 0.1, µ = 0.2 and a 10-qubit discretization (1024 points), with a time step δt = 0.01. In circles we show the theoretical predictions. 0 2 4 6 8 10 t 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 b) x = x 2 = (x x)2 x, theory 2, theory 10 5 0 5 10 0 2 4 6 8 10 a) 0.000 0.001 0.002 0.003 0.004 0.005 0.006 0.007 Figure 9: Solution of the Fokker-Planck equation using MPS and ﬁnite diﬀerences. (a) Density plot of the probability distribution evolved with the Fokker-Planck equation (69) and algorithm (73). In dashed lines we plot the expected value ¯x(t) and the curves ¯x(t) ± σ(t). (b) Center ¯x(t) and variance σ2(t) of the a probability distribution. We use D = 0.1, µ = 0.2 and a 10-qubit discretization (1024 points), with a time step δt = 0.01. In circles we show the theoretical predictions. 0 2 4 6 8 10 t 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 b) x = x 2 = (x x)2 x, theory 2, theory 0.000 0.001 0.002 0.003 0.004 0.005 0.006 0.007 Figure 9: Solution of the Fokker-Planck equation using MPS and ﬁnite diﬀerences. (a) Density plot of the probability distribution evolved with the Fokker-Planck equation (69) and algorithm (73). In dashed lines we plot the expected value ¯x(t) and the curves ¯x(t) ± σ(t). 5.4 Solving partial diﬀerential equations (b) Center ¯x(t) and variance σ2(t) of the a probability distribution. We use D = 0.1, µ = 0.2 and a 10-qubit discretization (1024 points), with a time step δt = 0.01. In circles we show the theoretical predictions. using uniform drift and diﬀusion, µ(x, t) = µ and D(x, t) = D. This equation governs the evolution of probability distributions for random variables undergoing a Wiener process in the Îto representation—a recurrent problem in quantum optics and ﬁnance, for instance. Moreover, this is already a challenging toy model from numerical analysis that is subject to numerical instabilities and demands state-of-the-art integration techniques. We will pro- vide two techniques to solve this equation using the MPS encoding of a quantum register, matching the two methods to work with diﬀerential operators from Sects. 5.3.1 and 5.3.2. 5.4.1 Finite diﬀerences Let us assume that we have the MPS representation of the initial value p(x, 0) and we need to estimate the evolution of this probability distribution at later times, p(x, t). Following Sect. 5.3.1, we write a ﬁnite-diﬀerence approximation to the Fokker-Planck model, replacing derivative operators with ladder operators. This transforms the Fokker-Planck equation into a ﬁrst order diﬀerential equation ∂t \|p(t)⟩= ˆG \|p(t)⟩, (70) (70) generated by the linear operator ˆG, whose action we approximate with an MPO. For concreteness, we will use the combined ﬁrst-second order approximation from (66) generated by the linear operator ˆG, whose action we approximate with an MPO. For concreteness, we will use the combined ﬁrst-second order approximation from (66) (66) ˆG = −µ ˆS+ −ˆS− 2δx + D ˆS+ + ˆS−−1 δx2 . (71) (71) We also need to build an implicit integration method that works around the fact that equa- tion (69) is not unitary and has the potential to develop exponentially growing numerical instabilities. We have chosen a second order implicit method, We also need to build an implicit integration method that works around the fact that equa- tion (69) is not unitary and has the potential to develop exponentially growing numerical instabilities. We have chosen a second order implicit method, ∂t \|p(x, t)⟩≃1 δt [\|p(x, t + δ)⟩−\|p(x, t)⟩] ≃1 2 ˆG [\|p(x, t + δ)⟩−\|p(x, t)⟩] . (72) (72) which translates into our integration recipe \|p(t + δt)⟩≃ 1 −1 2δt ˆG −1 1 + 1 2δt ˆG \|p(x, t)⟩. (73) (73) 24 Since ˆG has a simple representation in terms of MPO’s, we build our algorithm around the repetition of two elementary steps. First, compute the MPS for the product \|φ1(t)⟩= (1 + 1 2δt ˆG) \|p(x, t)⟩, using standard simpliﬁcation techniques to get the simplest and best approximation. Second, estimate the MPS \|φ2(t)⟩that best approximates the equation 1 −1 2δt ˆG \|φ2(t)⟩= \|φ1(t)⟩. (74) (74) In practice, we implement this step using a conjugate gradient method3, that only requires the matrix-vector multiplication between the MPO (1 −1 2δt ˆG) and a state represented in MPS form [27, 28]. In this form, a single evolution step has a cost O(Tcgs×SimpNm) given by the number of iterations in the conjugate-gradient method Tcgs times the cost of a single MPO-MPS mul- tiplication, which is dominated by the simpliﬁcation cost SimpNm ∼O(log2(1/εint)Nχ3). 5.4.1 Finite diﬀerences This cost has to be compared with the classical one, in which we represent the discretized function as a vector with 2Nm components and encode ˆG as a sparse matrix. In this case the conjugate-gradient method will take the same number of steps, but the matrix- vector multiplication cost can be exponentially larger, roughly proportional to the size of the vector O(2Nm) ∼O(ε−N int ), thanks to the sparse representation. Once more, if the MPS representation remains smooth and low-entangled, with bounded χ, there can be an exponential gain. As illustration, Fig. 9a shows the solution of the Fokker-Planck equation for a state that is initially Gaussian with variance σ(0) = 1 and center ¯x(0) = 0, living in an interval [−10, 10] discretized with m = 10 bits. The ﬁnite diﬀerence methods have some intrinsic limitations. The operator ∥δt ˆG∥∝22m diverges as we add more and more qubits. Our simulation is therefore very limited both in time and in the spatial discretization. This intrinsic instability is only partially cured with the implicit methods, but not always and also not for very long simulations. If we wish to have higher precision and numerical stability, we need to develop slightly better techniques, described in the following section. 3One could also write quadratic optimization techniques that minimize the distance between both states, as in DMRG’s correction vector techniques [37–39]. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.4.2 Spectral split-step method 0 10 20 30 t 10 6 10 5 error bound pexact p b) 100 steps 10 steps 10 5 0 5 x 0 5 10 15 20 25 30 t a) Figure 10: Solution of the Fokker-Planck equation using the Quantum Fourier Transform, Figure 10: Solution of the Fokker-Planck equation using the Quantum Fourier Transform, for D = 0.1, µ = 0.5 and a 14-qubit discretization (16384 points in space). (a) Density plot of the probability distribution. In dashed lines we plot the expected value ¯x(t) and the curves ¯x ± σ. Note how the QFT implements periodic boundary conditions and the wavefunction wraps around the borders. (b) Error bounds for a time-step δt = 0.3 (solid) and δt = 3. Note how the estimate decreases, indicating that our error bounds are pessimistic and that we can implement evolution for arbitrary long times without truncation errors. The function G(iˆk) quasi-momentum operator (53) is approximated by small MPO’s, as explained in Sect. 4.4. Unlike the ﬁnite diﬀerence method, if the generator G(∂x) does not depend on x, the solution (77) is exact: it works for all times in a single step. We have implemented the recipe (77) using MPS and MPO’s. We represent ˆF, ˆF−1 and exp(−µˆk + Dˆk2) using 3(m + 1) MPO’s of bond dimension 2. Provided that states remain weakly entangled, we can exactly solve the time evolution, without any numerical instabilities or truncation error, for any time and coeﬃcients. As regards the eﬃciency, the cost is dominated by the simpliﬁcation stages in the QFT and MPO-MPS product stages, which for the operators at hand seem have same practical asymptotic behavior O(Nm × SimpNm) ∼O(log2(1/εint)2N2χ3). In contrast, the cost of implementing this algorithm classically, using a traditional vector representation, scales linearly with the size of the discretized function. More precisely, the FFT [18] dominates the cost O(Nm2Nm) ∼ O(log2(1/εint)εN int), and we expect useful speedups for many well-behaved problems. As illustration, Fig. 10 reproduces once more the solution of the Fokker-Planck equation for a state that is initially Gaussian with variance σ(0) = 1 and center ¯x(0) = 0, living in an interval [−10σ, 10σ] discretized with m = 14 bits (16384 points in space). We can aﬀord larger number of qubits and longer times than in the ﬁnite diﬀerence method, because the algorithm is orders of magnitude more eﬃcient and very stable numerically. 5.4.2 Spectral split-step method The spectral methods, and in particular the Fourier transform and the split-step method techniques [19], have been traditionally used in many nonlinear Optics and quantum me- chanical problems, due to their eﬃciency, stability and accuracy. The method is optimally designed to solve equations of the form ∂tp(x, t) = G(∂x)p(x, t), (75) (75) where G(∂x) is a function of the diﬀerential operator ∂x and the coordinates are deﬁned over a regular interval. It works by moving to Fourier space, where the generator of the evolution is a function of the momentum k where G(∂x) is a function of the diﬀerential operator ∂x and the coordinates are deﬁned over a regular interval. It works by moving to Fourier space, where the generator of the evolution is a function of the momentum k ∂t˜p(x, t) = G(ik)˜p(x, t) ⇒˜p(x, t) = eG(ik)t˜p(x, 0). (76) (76) Here ˜p is the Fourier transform of the original function, ˜p = ˆFp, over the real line. ere ˜p is the Fourier transform of the original function, ˜p = ˆFp, over the real line. In our discrete scenario with uniformly sampled, regular intervals, the spectral method can be implemented using the Quantum Fourier Transform (49) and the interpolation techniques from Sect. 5.3.2. The approximate solution to (75) is expressed as \|p(m)(t)⟩= ˆF−1 exp h G(iˆk)t i ˆF \|p(m)(0)⟩, (77) (77) 3One could also write quadratic optimization techniques that minimize the distance between both states, as in DMRG’s correction vector techniques [37–39]. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 25 0 10 20 30 t 10 6 10 5 error bound pexact p b) 100 steps 10 steps 10 5 0 5 x 0 5 10 15 20 25 30 t a) Figure 10: Solution of the Fokker-Planck equation using the Quantum Fourier Transform, for D = 0.1, µ = 0.5 and a 14-qubit discretization (16384 points in space). (a) Density plot of the probability distribution. In dashed lines we plot the expected value ¯x(t) and the curves ¯x ± σ. Note how the QFT implements periodic boundary conditions and the wavefunction wraps around the borders. (b) Error bounds for a time-step δt = 0.3 (solid) and δt = 3. Note how the estimate decreases, indicating that our error bounds are pessimistic and that we can implement evolution for arbitrary long times without truncation errors. 5.4.2 Spectral split-step method Notice also how the algorithm implements periodic boundaries by default; this is a feature that is very useful to avoid boundary reﬂections and simulate in small intervals the dynamics of propagating ﬁelds. This recipe can be extended to problems that include dependencies on both the spatial derivatives and the spatial coordinates. We explicitly refer to equations of the form (78) ∂tp(x, t) = [G(∂x) + V (x)] p(x, t), (78) or higher-dimensional equivalents. The solution to this problem is no longer exact, but or higher-dimensional equivalents. The solution to this problem is no longer exact, but Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 26 Trotter-Suzuki expansion of the generator relies on a Trotter-Suzuki expansion of the generator relies on a Trotter-Suzuki expansion of the generator \|p(m)(t + δt)⟩= e[G(∂x)+V (x)]δt \|p(m)(t)⟩ ≃eV (x)δt/2 ˆF−1eG(iˆk)δt ˆFeV (x)δt/2 \|p(m)(t)⟩+ O(δt2), (79) (79) This technique is known as a split-step method because it combines evolution steps in real space, with stages that are implemented in Fourier space [19]. All ingredients in this formula (79) are well known from earlier pages. 4When there is a lot of squeezing, or when the interval size \|b−a\| is orders of magnitude larger than the exponential width, the computer cannot represent the extreme values that appear in the MPO tensors (47). Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.5 Construction of the MPS The algorithms from previous sections assume that we already have an MPS wavefunction to begin with. We will now discuss diﬀerent ways in which to obtain such initial conditions. Exact discretization The trivial way to construct an MPS representation is to begin with the discretized function p(m)(s1, s2 . . . sm) and perform a sequential Schmidt decom- position. This has an exponential cost O(23m/2) and fails when the number of qubits goes above 30-something, due to memory and time constraints, as we have seen in Sect. 3.6. Explicit formulas Some probability distributions admit analytical expressions. We can see those functions p(x) as a product p(x)e(x) with the uniform distribution e(x), an operation that can be approximated using the techniques from Sect. 4.5. An important example is the Gaussian probability distribution. When we express the coordinates in terms of qubits, the Gaussian (34) becomes the partition function of an Ising model H Ising model H p(xs1, xs2 . . . xsN ) = 1 Z(β)e−βH(s1 1,s1 2,...,sm N ) \|s1⟩. (80) (80) As discussed in Sect. 4.5, we could write the whole MPS using the exponential of a QUBO formula. In practice, this only works well when the state Σ is not very squeezed4. More generally, we need to reconstruct the whole probability state a progressive reﬁnement of the uniform distribution p(x) = 1 Z(β/K)e−βH/K K e(x), (81) (81) with each step implemented by an MPO with each step implemented by an MPO \|p⟩= ˆZMPO(β/K) · ˆZMPO(β/K) · · · ˆZMPO(β/K) \|e⟩, (82) (82) that is a better behaved Gaussian function. We have used this technique to reconstruct the MPS representations of the 2D (36) and 3D (37) Gaussian states. Figs. 3b and 4a show the maximum entanglement entropy over all bipartitions of the resulting MPS state, while Table 2 discusses the amount of memory and the size of the tensors. 4When there is a lot of squeezing, or when the interval size \|b−a\| is orders of magnitude larger than the exponential width, the computer cannot represent the extreme values that appear in the MPO tensors (47). 27 Imaginary time evolution Equation (82) is nothing but a discrete approximation to the imaginary time evolution of the unnormalized state \|pu⟩ ∂β \|pu⟩= −ˆH \|pu⟩, (83) (83) with uniform initial condition \|pu(β = 0)⟩= \|e⟩. 5.5 Construction of the MPS This equation can be solved for any partition function distribution (80) generated by an operator ˆH with an eﬃcient MPO representation. This approach is reminiscent of how thermal state density matrix have been simulated using the MPS formalism [29], but it is much simpler, since we do not introduce any auxiliary puriﬁcation degrees of freedom. Machine learning The challenge of writing an MPS representation of a function p(x) is comparable to the challenge of constructing that function in a quantum computer: there are eﬃcient protocols, but we do not know them a priori. The quantum computing ﬁeld has developed diﬀerent strategies to address this problem. A promising one is to approximate the state \|p⟩using a parameterized quantum circuit that is trained using the techniques of machine learning, such as generative adversarial networks [40]. These techniques can be extended to our domain. Instead of using a quantum-classical approach with a circuit that generates the probabilities and a neural network that discriminates, we can use the MPS as generator and apply similar training techniques. 5.6 Porting back to the quantum computers This work has introduced various classical MPS algorithms that use the tools of quan- tum computing—quantum register representation, function encodings, quantum gates and algorithms—to solve eﬃciently various numerical analysis tasks. These techniques, and the whole line of research, can feed back to the world of quantum computing. For instance, the Fourier interpolation algorithm from Sect. 5.2.2 has an immediate translation to a real quantum computer. Other algorithms require some ﬁne tuning. In linear interpolation and ﬁnite diﬀerences, the the ˆS± operator was designed to use irreversible arithmetic. However, we know that a similar operator can be implemented or approximated using auxiliary qubits and reversible arithmetic—half and full adders—, or other ideas from quantum simulation [8]. Something similar happens in the case of PDE solvers. Our discussion has focused on the non-unitary evolution induced by Fokker-Planck techniques, studying the implemen- tation of the non-unitary operator exp[G(ˆk)]. Algorithms (77) and (79) can be trivially generalized to solve Schrödinger equations such as ∂tψ(x, t) = −i ℏ 2m∇2 −iV (x)/ℏ ψ(x, t). (84) (84) The generator of this equation is anti-Hermitian and both exp[δt G(iˆk)] and exp[−iV (x)δt/ℏ] can be implemented as a unitary gate in the quantum register. As in the MPS case, the scaling of the algorithm is problem dependent. The exponential exp[iαˆk2] can be imple- mented with O(N2m2) steps, but the exponential of V (x) may have a more complicated scaling, strongly dependent on function arithmetic. However, we expect that the smooth- ness of usual potentials will also lead to simple approximations with quasi-local gates. The generator of this equation is anti-Hermitian and both exp[δt G(iˆk)] and exp[−iV (x)δt/ℏ] can be implemented as a unitary gate in the quantum register. As in the MPS case, the scaling of the algorithm is problem dependent. The exponential exp[iαˆk2] can be imple- mented with O(N2m2) steps, but the exponential of V (x) may have a more complicated scaling, strongly dependent on function arithmetic. However, we expect that the smooth- ness of usual potentials will also lead to simple approximations with quasi-local gates. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 5.7 Implementation Together with this manuscript, we have developed an accompanying MPS library [27] and supplementary driving code [28], all of which are available as separate Zenodo/GitHub Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 28 repositories. The SeeMPS library [27] contains an implementation of all MPS fundamen- tal algorithms, including the fundamental representation, approximation of linear combi- nations of states, MPO-MPS contractions, conjugate-gradient for matrix inversion, and Quantum Fourier Transform. The library is written using the literate-programming ap- proach, and combines Python code with the mathematical formulation of the algorithms that we have explained. The supplementary material to this manuscript [28] is also available as Zenodo/GitHub respository. It builds on the given version of the SeeMPS library, using it to implement all methods and simulations in this work . In particular, it shows (i) how to construct the quantum-register approximations of the probability distributions studied in Sect. 3.5, (ii) eﬃcient techniques to construct Gaussian multivariate probability distributions using MPS, (iii) a study of Fourier interpolation with MPS, (iv) a program to solve the Fokker- Planck using the ﬁnite-diﬀerence and conjugate-gradient methods, and (v) a solution of the Fokker-Planck equation using the MPS split-step method. Those codes are the ones that have been used to produce the plots in this work. 6 Discussion and outlook This work has presented many numerical algorithms for constructing, manipulating and interrogating multivariate functions in MPS-simulated quantum registers. We have shown that, heuristically, the renormalization provided by the quantum register representation is key to the creation of states with low entanglement, capable of encoding smooth, diﬀeren- tiable functions. The use of tensor network states is a modern development in numerical analysis [14, 15]. In the ﬁeld of low-rank tensor approximations, a multivariate function ψ(x1, x2 . . . xN) is approximated by a contraction of tensors, labeled either by the continuous variables xi, or discretized versions of them xsi. In other works, the spatial degrees of freedom are replaced by labels in some local mode expansion [16]. These approaches use a Hilbert space decomposition that preserves the notion of local degrees of freedom, by expanding a function in a normal basis of modes f(x1, x2, . . . , xN) = X c(i1, i2, . . . , iN)φi1(x1)φi2(x2) · · · φiN (xN), (85) (85) and using a low-rank tensor approximation to represent c(i1, i2, . . . , iN). It can be argued that such encodings are isomorphic to the A-order tensor structure discussed in Section 3.6. Since the orthonormal modes φin(xn) can be expanded in the basis of m qubits that represents the n-th coordinate, \|s1 n, . . . , sm n ⟩, these modes can be regarded as an isometry from the space of qubits {s1 n, . . . , sm n } to the mode coordinate {in}. This isometry cannot cure the fact that the entanglement between coordinates grows in an unfavorable way with the dimensionality of the problem, as we saw in Section 3.6. This entanglement growth will aﬀect the complexity of any splitting of the tensor c(i1, i2, . . . , iN) just as badly as it aﬀected our MPS representation. In this work we have presented a quantum register discretization that solves those limitations. In this representation each local coordinate is exploded into m qubits of information, each of them probing the function at a diﬀerent length scale. As we have seen in Sect. 3.6, these qubits need not be kept together, and there may be more eﬃcient renormalization schemes when we group common length scales that are more strongly correlated. Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. 6 Discussion and outlook Returning to the mode representation mentioned before, this technique of 29 Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. grouping qubits is comparable to a multi-scale wavelet modal representations f(x1, x2, . . . , xN) = X c(i1, i2, . . . , iN) Y n φin(x1, x2, . . . , xN), (86) grouping qubits is comparable to a multi-scale wavelet modal representations f(x1, x2, . . . , xN) = X c(i1, i2, . . . , iN) Y n φin(x1, x2, . . . , xN), (86) (86) where where each φin(x1, x2, . . . , xN) represents the oscillations of the function at a diﬀer- ent length scale, and our tensor c(i1, . . . , iN) explores correlations between diﬀerent length scales. This implicit renormalization scheme is a discovery of the quantum computing com- munity that has not been suﬃciently exploited so far and which may empower many recent developments in the ﬁeld of quantum ﬁnance and probability distribution analysis [4, 5, 26]. The algorithms in this paper bend themselves to a broad family of problems which have been considered in the quantum computing world. We have illustrated the solution of time-dependent partial diﬀerential equations, but the same techniques can be extended to stationary problems. This way, the MPS-simulated quantum register becomes a natural tool to solve the Poisson equation [8], the wave equation [41], the ﬂuid equations [35], or even the Schrödinger equation itself. This implies not just abstract, fundamental studies in Physics, but practical applications in ﬁelds such as aerodynamics or ﬁnance. We expect new applications of quantum-inspired ﬁnance that reach beyond the state-of-the-art [26], providing new schemes for evaluating ﬁnancial products [5], performing risk analysis [4] and even more sophisticated time-dependent simulations and tracking. Let us also remark that the algorithms developed in this work are of a heuristic nature. All methods and techniques in Sections 4 and 5 assume states with low entanglement. This approximation is bound to break at some point, either because of functions with broad spectra and complex structure, or because of increased dimensionality. Quantum computers become a valuable tool that still has an exponential advantage over classical algorithms, and which may proﬁt from the ideas and developments associated to MPS- simulated quantum register techniques, as discussed before [cf. Sect. 5.6]. 6 Discussion and outlook Finally, all simulations in this paper have been implemented with the SeeMPS Python library [27, 28]. While Python is a slow language, the algorithms are high-level and beneﬁt from the eﬃcient tensor operations that are available in Numpy and Scipy. This makes the code performant enough for practical applications. However, it is undeniable that recent frameworks based on TensorFlow [42] and highly parallelized architectures for tensor contractions [43] would be a better platform to real-world deployment. J.J.G.-R. thanks Andrea Cadarso for discussions around the motivation of the method and its applications. This material is based upon work supported by the Air Force Oﬃce of Scientiﬁc Research under award number FA2386-18-1-4019. This work has been sup- ported by funding from project PGC2018-094792-B-I00 (MCIU/AEI/FEDER, UE) and and CAM/FEDER Project No. S2018/TCS-4342 (QUITEMAD-CM). Accepted in Quantum 2020-04-12, click title to verify. Published under CC-BY 4.0. References [1] C. Zalka. Simulating quantum systems on a quantum computer. 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https://openalex.org/W2297338717	OpenAlex	Open Science	CC-By	2,009	Bis[5-(2-naphthyl)-1<i>H</i>-pyrazole-κ<i>N</i><sup>2</sup>]silver(I) nitrate	Zhaoyang Wang	Latin	Spoken	2,121	6,919	Zhao-Yang Wang,a Pei-Pei Zhang,a Guang Yanga and Seik Weng Ngb* Zhao-Yang Wang,a Pei-Pei Zhang,a Guang Yanga and Seik Weng Ngb* 2268 measured reﬂections 2056 independent reﬂections 1752 reﬂections with I > 2(I) Rint = 0.061 aDepartment of Chemistry, Zhengzhou University, Zhengzhou 450001, People’s Republic of China, and bDepartment of Chemistry, University of Malaya, 50603 Kuala Lumpur, Malaysia C d il ik @ d H-atom parameters constrained max = 0.36 e A˚ 3 min = 0.48 e A˚ 3 Absolute structure: Flack (1983), 353 Friedel pairs Flack parameter: 0.33 (8) Received 20 July 2009; accepted 20 July 2009 Key indicators: single-crystal X-ray study; T = 291 K; mean (C–C) = 0.019 A˚; disorder in main residue; R factor = 0.061; wR factor = 0.174; data-to-parameter ratio = 11.5. The Ag atom in the title compound, [Ag(C13H10N2)2]NO3, shows an approximately linear coordination [N–Ag–N 162.6 (4)]. The coordination geometry is distorted towards square-planar owing to two long Ag O interactions [Ag O = 2.634 (15) and 2.861 (13) A˚ ]. In the crystal structure, the Ag atom lies on a special position of 2 site symmetry; the nitrate anion is disordered about the special position. The crystal under investigation was a racemic twin with a 33% minor twin component. Data collection: RAPID-AUTO (Rigaku, 1998); cell reﬁnement: RAPID-AUTO; data reduction: CrystalStructure (Rigaku/MSC, 2002); program(s) used to solve structure: SHELXS97 (Sheldrick, 2008); program(s) used to reﬁne structure: SHELXL97 (Sheldrick, 2008); molecular graphics: X-SEED (Barbour, 2001); software used to prepare material for publication: publCIF (Westrip, 2009). We thank the Education Department of Henan Province, Zhengzhou University and the University of Malaya for supporting this study. metal-organic compounds Related literature Supplementary data and ﬁgures for this paper are available from the IUCr electronic archives (Reference: XU2561). This structure is the ﬁrst report of a metal complex of the 5-(2- naphthyl)-1H-pyrazole; for the synthesis of this N-heterocycle, see: Yang & Raptis (2003). metal-organic compounds Experimental Crystal data [Ag(C13H10N2)2]NO3 Mr = 558.34 Monoclinic, C2 a = 13.911 (6) A˚ b = 7.340 (1) A˚ c = 12.669 (5) A˚ = 113.43 (2) V = 1186.9 (7) A˚ 3 Z = 2 Mo K radiation = 0.89 mm1 T = 291 K 0.20 0.18 0.16 mm Data collection Rigaku R-AXIS RAPID IP diffractometer Absorption correction: multi-scan (ABSCOR; Higashi, 1995) Tmin = 0.843, Tmax = 0.871 2268 measured reﬂections 2056 independent reﬂections 1752 reﬂections with I > 2(I) Rint = 0.061 Reﬁnement R[F 2 > 2(F 2)] = 0.061 wR(F 2) = 0.174 S = 1.09 2056 reﬂections 179 parameters 32 restraints H-atom parameters constrained max = 0.36 e A˚ 3 min = 0.48 e A˚ 3 Absolute structure: Flack (1983), 353 Friedel pairs Flack parameter: 0.33 (8) Acta Crystallographica Section E Structure Reports Online ISSN 1600-5368 Acta Crystallographica Section E Structure Reports Online ISSN 1600-5368 Barbour, L. J. (2001). J. Supramol. Chem. 1, 189–191. Flack, H. D. (1983). Acta Cryst. A39, 876–881. Higashi, T. (1995). ABSCOR. Rigaku Corporation, Tokyo, Japan. Rigaku (1998). RAPID-AUTO. Rigaku Corporation, Tokyo, Japan. Rigaku/MSC (2002). CrystalStructure. Rigaku/MSC Inc., The Woodlands, Texas, USA. Sheldrick, G. M. (2008). Acta Cryst. A64, 112–122. Westrip, S. P. (2009). publCIF. In preparation. Yang, G. & Raptis, R. G. (2003). J. Heterocycl. Chem. 40, 659–664. S1. Experimental 3-(2-Naphthyl)pyrazole was prepared according to the literature method (Yang & Raptis, 2003). An acetonitrile solution (2 ml) of silver nitrate (0.02 mmol, 3.4 mg) was mixed with an ethanol solution (1 ml) of 3-(2-naphthyl)pyrazole (0.02 mmol, 4 mg). The pH value of the mixture was adjusted to about 5 by dilute nitric acid. The resulting solution was allowed to evaporate for two weeks to yield colorless crystals in 60% yield. References m980 Wang et al. Acta Cryst. (2009). E65, m980 doi:10.1107/S1600536809028645 supporting information S2. Refinement The measurements are complete to 94% at a 2θ of 55 °, but are complete to 98% at a 2θ of 50 °. The nitrate group is disordered about the twofold rotation axis; the anion was allowed to refine off the symmetry element. The three N–O distances were restrained to within 0.01 Å of each other, as were the three O···O distances. The four atoms were restrained to be nearly planar, and their anisotropic temperature factors were restrained to be nearly isotropic. Carbon-bound H atoms were placed in calculated positions [C—H = 0.93, N–H 0.89 Å; U(H) = 1.2Ueq(C,N)]. The crystal under investigation is a racemic twin; the explicit refinement of the Flack parameter gave a minor twin t f 33% The crystal under investigation is a racemic twin; the explicit refinement of the Flack parameter gave a minor twin component of 33%. Figure 1 Thermal ellipsoid plot of the [Ag(C13H10N2)2][NO3], displacement ellipsoids are drawn at the 50% probability level. The nitrate anion is disordered about a special position; the disorder is not shown. Acta Cryst. (2009). E65, m980 [doi:10.1107/S1600536809028645] Acta Cryst. (2009). E65, m980 [doi:10.1107/S1600536809028645] Figure 1 Thermal ellipsoid plot of the [Ag(C13H10N2)2][NO3], displacement ellipsoids are drawn at the 50% probability level. The nitrate anion is disordered about a special position; the disorder is not shown. Acta Cryst. (2009). E65, m980 sup-1 supporting information Bis[5-(2-naphthyl)-1H-pyrazole-κN2]silver(I) n Crystal data [Ag(C13H10N2)2]NO3 Mr = 558.34 Monoclinic, C2 Hall symbol: C 2y a = 13.911 (6) Å b = 7.340 (1) Å c = 12.669 (5) Å β = 113.43 (2)° V = 1186.9 (7) Å3 Z = 2 Data collection Rigaku R-AXIS RAPID IP diffractometer Radiation source: fine-focus sealed tube Graphite monochromator ω scans Absorption correction: multi-scan (ABSCOR; Higashi, 1995) Tmin = 0.843, Tmax = 0.871 Refinement Refinement on F2 Least-squares matrix: full R[F2 > 2σ(F2)] = 0.061 wR(F2) = 0.174 S = 1.09 2056 reflections 179 parameters 32 restraints Primary atom site location: structure-invariant direct methods Secondary atom site location: difference Fourier map Bis[5-(2-naphthyl)-1H-pyrazole-κN2]silver(I) nitrate F(000) = 564 Dx = 1.562 Mg m−3 Mo Kα radiation, λ = 0.71073 Å Cell parameters from 2056 reflections θ = 1.8–27.5° µ = 0.89 mm−1 T = 291 K Block, colorless 0.20 × 0.18 × 0.16 mm 2268 measured reflections 2056 independent reflections 1752 reflections with I > 2σ(I) Rint = 0.061 θmax = 27.5°, θmin = 1.8° h = −17→8 k = −9→7 l = −14→16 Hydrogen site location: inferred from neighbouring sites H-atom parameters constrained w = 1/[σ2(Fo2) + (0.1009P)2 + 2.0218P] where P = (Fo2 + 2Fc2)/3 (Δ/σ)max = 0.001 Δρmax = 0.36 e Å−3 Δρmin = −0.48 e Å−3 Extinction correction: SHELXL97 (Sheldrick, 2008), Fc=kFc[1+0.001xFc2λ3/sin(2θ)]-1/4 Extinction coefficient: 0.010 (2) Absolute structure: Flack (1983), 353 Friedel pairs Absolute structure parameter: 0.33 (8) Hydrogen site location: inferred from neighbouring sites Hydrogen site location: inferred from neighbouring sites H-atom parameters constrained ρmin Extinction correction: SHELXL97 (Sheldrick, 2008), Fc=kFc[1+0.001xFc2λ3/sin(2θ)]-1/4 Extinction coefficient: 0.010 (2) Absolute structure: Flack (1983), 353 Friedel pairs Absolute structure parameter: 0.33 (8) Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) x y z Uiso/Ueq Occ. Figure 1 (<1) Ag1 0.5000 0.5000 (2) 0.5000 0.0696 (4) O1 0.5305 (13) 0.8398 (18) 0.4537 (12) 0.079 (4) 0.50 O2 0.4805 (13) 1.1184 (16) 0.4497 (12) 0.078 (4) 0.50 O3 0.4805 (13) 0.9378 (17) 0.5844 (11) 0.079 (4) 0.50 N1 0.6133 (6) 0.4563 (10) 0.4293 (6) 0.060 (2) N2 0.6235 (6) 0.2913 (10) 0.3832 (6) 0.0565 (16) H2 0.5892 0.1907 0.3868 0.068 N3 0.4971 (9) 0.9654 (11) 0.4961 (9) 0.062 (3) 0.50 C1 0.6779 (7) 0.5684 (15) 0.4085 (8) 0.068 (2) H1 0.6879 0.6898 0.4312 0.082* Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) sup-2 Acta Cryst. (2009). Figure 1 E65, m980 supporting information supporting information C2 0.7299 (7) 0.479 (3) 0.3471 (7) 0.068 (3) H2A 0.7788 0.5297 0.3225 0.082* C3 0.6928 (7) 0.3009 (12) 0.3311 (7) 0.0546 (18) C4 0.7151 (6) 0.1443 (12) 0.2722 (7) 0.0537 (18) C5 0.6475 (6) −0.008 (3) 0.2316 (6) 0.0616 (18) H5 0.5874 −0.0157 0.2467 0.074* C6 0.6685 (8) −0.1416 (16) 0.1720 (8) 0.070 (2) H6 0.6211 −0.2367 0.1431 0.084* C7 0.7620 (8) −0.1405 (17) 0.1522 (8) 0.067 (2) C8 0.7883 (9) −0.282 (2) 0.0913 (9) 0.080 (3) H8 0.7440 −0.3821 0.0639 0.096* C9 0.8788 (10) −0.271 (2) 0.0733 (9) 0.096 (4) H9 0.8948 −0.3637 0.0328 0.115* C10 0.9456 (8) −0.130 (3) 0.1125 (10) 0.094 (4) H10 1.0069 −0.1275 0.0997 0.113* C11 0.9243 (6) 0.009 (4) 0.1703 (6) 0.082 (3) H11 0.9719 0.1039 0.1966 0.098* C12 0.8311 (6) 0.014 (3) 0.1924 (6) 0.060 (2) C13 0.8064 (6) 0.1497 (16) 0.2520 (8) 0.063 (2) H13 0.8519 0.2479 0.2795 0.075* Atomic displacement parameters (Å2) U11 U22 U33 U12 U13 U23 Ag1 0.0884 (6) 0.0661 (6) 0.0753 (6) 0.000 0.0547 (5) 0.000 O1 0.077 (7) 0.070 (8) 0.097 (8) 0.012 (6) 0.043 (6) −0.013 (7) O2 0.089 (8) 0.058 (7) 0.093 (8) 0.009 (6) 0.043 (7) 0.006 (6) O3 0.085 (7) 0.088 (9) 0.076 (7) 0.007 (6) 0.045 (6) 0.005 (6) N1 0.076 (4) 0.052 (6) 0.062 (4) 0.002 (3) 0.039 (3) −0.003 (3) N2 0.072 (4) 0.050 (4) 0.056 (4) −0.001 (3) 0.034 (4) −0.004 (3) N3 0.054 (4) 0.071 (9) 0.067 (5) 0.001 (10) 0.029 (4) −0.005 (10) C1 0.071 (5) 0.074 (6) 0.060 (5) −0.009 (4) 0.027 (4) −0.003 (4) C2 0.068 (4) 0.072 (9) 0.068 (4) −0.003 (6) 0.033 (3) 0.001 (6) C3 0.060 (4) 0.057 (4) 0.054 (4) 0.001 (3) 0.029 (4) 0.004 (3) C4 0.055 (4) 0.053 (4) 0.058 (5) 0.005 (3) 0.028 (4) 0.009 (4) C5 0.071 (4) 0.060 (5) 0.067 (4) 0.008 (7) 0.041 (3) 0.006 (8) C6 0.075 (5) 0.074 (6) 0.064 (6) −0.004 (5) 0.031 (5) 0.002 (5) C7 0.077 (5) 0.085 (7) 0.048 (5) 0.008 (5) 0.034 (4) 0.004 (4) C8 0.090 (6) 0.089 (8) 0.068 (5) 0.011 (6) 0.037 (5) −0.017 (6) C9 0.108 (9) 0.124 (11) 0.065 (6) 0.030 (8) 0.043 (6) −0.015 (7) C10 0.053 (5) 0.160 (14) 0.066 (6) 0.022 (6) 0.019 (5) −0.023 (7) C11 0.053 (4) 0.141 (10) 0.055 (4) 0.004 (10) 0.024 (3) −0.018 (11) C12 0.056 (3) 0.081 (7) 0.044 (3) 0.002 (7) 0.021 (3) 0.003 (7) C13 0.051 (4) 0.091 (7) 0.056 (5) −0.001 (4) 0.032 (4) −0.004 (4) Atomic displacement parameters (Å2) sup-3 Acta Cryst. Figure 1 (2009). E65, m980 supporting information Geometric parameters (Å, º) Ag1—N1i 2.124 (7) C4—C5 1.42 (2) Ag1—N1 2.124 (7) C5—C6 1.34 (2) Ag1—O1 2.634 (15) C5—H5 0.9300 Ag1—O2ii 2.861 (13) C6—C7 1.419 (13) O1—N3 1.246 (10) C6—H6 0.9300 O2—N3 1.245 (10) C7—C8 1.427 (15) O3—N3 1.246 (10) C7—C12 1.44 (2) N1—C1 1.319 (12) C8—C9 1.369 (17) N1—N2 1.375 (10) C8—H8 0.9300 N2—C3 1.370 (11) C9—C10 1.35 (2) N2—H2 0.8900 C9—H9 0.9300 C1—C2 1.416 (16) C10—C11 1.35 (3) C1—H1 0.9300 C10—H10 0.9300 C2—C3 1.39 (2) C11—C12 1.432 (10) C2—H2A 0.9300 C11—H11 0.9300 C3—C4 1.469 (12) C12—C13 1.38 (2) C4—C13 1.393 (11) C13—H13 0.9300 N1i—Ag1—N1 162.6 (4) C6—C5—C4 121.3 (7) N1i—Ag1—O1 116.7 (4) C6—C5—H5 119.4 N1—Ag1—O1 80.6 (4) C4—C5—H5 119.4 N1i—Ag1—O2ii 85.7 (4) C5—C6—C7 121.2 (10) N1—Ag1—O2ii 77.2 (4) C5—C6—H6 119.4 O1—Ag1—O2ii 152.4 (5) C7—C6—H6 119.4 N3—O1—Ag1 119.0 (7) C6—C7—C8 122.8 (11) C1—N1—N2 105.7 (7) C6—C7—C12 118.1 (10) C1—N1—Ag1 131.9 (7) C8—C7—C12 119.1 (9) N2—N1—Ag1 122.0 (5) C9—C8—C7 119.8 (12) C3—N2—N1 111.7 (7) C9—C8—H8 120.1 C3—N2—H2 124.1 C7—C8—H8 120.1 N1—N2—H2 124.1 C10—C9—C8 122.0 (11) O2—N3—O1 119.7 (7) C10—C9—H9 119.0 O2—N3—O3 120.3 (7) C8—C9—H9 119.0 O1—N3—O3 120.0 (7) C11—C10—C9 120.7 (11) N1—C1—C2 111.1 (12) C11—C10—H10 119.6 N1—C1—H1 124.5 C9—C10—H10 119.6 C2—C1—H1 124.5 C10—C11—C12 122 (2) C3—C2—C1 105.8 (9) C10—C11—H11 118.8 C3—C2—H2A 127.1 C12—C11—H11 118.8 C1—C2—H2A 127.1 C13—C12—C11 124.7 (18) C2—C3—N2 105.6 (7) C13—C12—C7 119.2 (7) C2—C3—C4 132.0 (7) C11—C12—C7 116.1 (17) N2—C3—C4 122.4 (8) C12—C13—C4 121.7 (10) C13—C4—C5 118.4 (8) C12—C13—H13 119.2 C13—C4—C3 117.6 (8) C4—C13—H13 119.2 C5—C4—C3 123.9 (8) sup-4 Acta Cryst. (2009). Figure 1 E65, m980 supporting information N1i—Ag1—O1—N3 15.5 (10) C2—C3—C4—C5 −158.6 (11) N1—Ag1—O1—N3 −166.2 (9) N2—C3—C4—C5 20.6 (14) O2ii—Ag1—O1—N3 157.0 (11) C13—C4—C5—C6 −2.1 (16) N1i—Ag1—N1—C1 −170.0 (8) C3—C4—C5—C6 175.7 (10) O1—Ag1—N1—C1 15.2 (9) C4—C5—C6—C7 3.3 (17) O2ii—Ag1—N1—C1 178.7 (9) C5—C6—C7—C8 178.5 (11) N1i—Ag1—N1—N2 17.3 (6) C5—C6—C7—C12 −3.4 (15) O1—Ag1—N1—N2 −157.4 (8) C6—C7—C8—C9 178.8 (10) O2ii—Ag1—N1—N2 6.0 (6) C12—C7—C8—C9 0.8 (16) C1—N1—N2—C3 −1.4 (10) C7—C8—C9—C10 0.7 (19) Ag1—N1—N2—C3 173.0 (5) C8—C9—C10—C11 −1 (2) Ag1—O1—N3—O2 −157.6 (9) C9—C10—C11—C12 0 (2) Ag1—O1—N3—O3 22.4 (10) C10—C11—C12—C13 179.1 (13) N2—N1—C1—C2 0.9 (10) C10—C11—C12—C7 2 (2) Ag1—N1—C1—C2 −172.6 (6) C6—C7—C12—C13 2.5 (16) N1—C1—C2—C3 −0.2 (11) C8—C7—C12—C13 −179.5 (11) C1—C2—C3—N2 −0.7 (10) C6—C7—C12—C11 −179.9 (11) C1—C2—C3—C4 178.6 (9) C8—C7—C12—C11 −1.8 (16) N1—N2—C3—C2 1.3 (10) C11—C12—C13—C4 −178.8 (12) N1—N2—C3—C4 −178.1 (8) C7—C12—C13—C4 −1.4 (17) C2—C3—C4—C13 19.2 (15) C5—C4—C13—C12 1.2 (15) N2—C3—C4—C13 −161.6 (8) C3—C4—C13—C12 −176.7 (10) S t d (i) +1 +1 (ii) 1 Hydrogen-bond geometry (Å, º) D—H···A D—H H···A D···A D—H···A N2—H2···O2ii 0.89 2.04 2.76 (2) 137 N2—H2···O3iii 0.89 2.19 3.08 (2) 173 Symmetry codes: (ii) x, y−1, z; (iii) −x+1, y−1, −z+1. Symmetry codes: (ii) x, y−1, z; (iii) −x+1, y−1, −z+1. sup-5 Acta Cryst. (2009). E65, m980	3,251
https://openalex.org/W4230876385	OpenAlex	Open Science	CC-By	2,012	The accommodation coefficient of water molecules on ice-cirrus cloud studies at the AIDA simulation chamber	J. Skrotzki	English	Spoken	15,299	27,043	E th S nvited contribution by J. Skrotzki, recipient of the EGU Union Outstanding Student Poster Award 20 Correspondence to: H. Saathoff (harald.saathoff@kit.edu) Received: 29 June 2012 – Published in Atmos. Chem. Phys. Discuss.: 18 September 2012 Revised: 25 March 2013 – Accepted: 3 April 2013 – Published: 29 April 2013 Abstract. Cirrus clouds and their impact on the Earth’s ra- diative budget are subjects of current research. The processes governing the growth of cirrus ice particles are central to the radiative properties of cirrus clouds. At temperatures rele- vant to cirrus clouds, the growth of ice crystals smaller than a few microns in size is strongly inﬂuenced by the accom- modation coefﬁcient of water molecules on ice, αice, making this parameter relevant for cirrus cloud modeling. However, the experimentally determined magnitude of αice for cirrus temperatures is afﬂicted with uncertainties of almost three orders of magnitude, and values for αice derived from cir- rus cloud data lack signiﬁcance so far. This has motivated dedicated experiments at the cloud chamber AIDA (Aerosol Interactions and Dynamics in the Atmosphere) to determine αice in the cirrus-relevant temperature interval between 190 K and 235 K under realistic cirrus ice particle growth condi- tions. The experimental data sets have been evaluated in- dependently with two model approaches: the ﬁrst relying on the newly developed model SIGMA (Simple Ice Growth Model for determining Alpha), the second one on an estab- lished model, ACPIM (Aerosol-Cloud-Precipitation Interac- tion Model). Within both approaches a careful uncertainty analysis of the obtained αice values has been carried out for each AIDA experiment. The results show no signiﬁcant de- pendence of αice on temperature between 190 K and 235 K. In addition, we ﬁnd no evidence for a dependence of αice Instrumentation Methods and Data Systems en Access Geoscientific Model Development Open Access Hydrology and O on ice particle size or on water vapor supersaturation for ice particles smaller than 20 µm and supersaturations of up to 70 %. The temperature-averaged and combined result from both models is αice = 0.7+0.3 −0.5, which implies that αice may only exert a minor impact on cirrus clouds and their charac- teristics when compared to the assumption of αice = 1. Im- pact on prior calculations of cirrus cloud properties, e.g., in climate models, with αice typically chosen in the range 0.2–1 is thus expected to be negligible. In any case, we provide a well-constrained αice which future cirrus model studies can rely on. 1 Introduction Ocean Science Open Access Solid Earth Open Access Cirrus clouds play a major role in the radiative budget of the Earth’s atmospheric system through their interactions with incident solar and surface-emitted terrestrial radiation (Liou, 1986). The radiative properties of cirrus clouds strongly de- pend on ice particle size, shape, and number concentration (Zhang et al., 1999). Ice particle properties and number con- centration in cirrus clouds depend, besides other inﬂuences, on ice particle growth rates (Lin et al., 2002). One of the main parameters governing the growth of ice particles up to a size of few micrometers, i.e., in the initial stage of ice particle Atmospheric Chemistry and Physics Open Access Atmospheric Chemistry and Physics Open Access Cli t Ope 1Institute for Meteorology and Climate Research – Atmospheric Aerosol Research, Karlsruhe Institute of Technology (KIT), Karlsruhe Germany Climate Ope 1Institute for Meteorology and Climate Research – Atmospheric Aerosol Research, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany Clim of the P 2School of Earth, Atmospheric and Environmental Sciences, University of Manchester, Manchester, UK 3Physikalisch-Technische Bundesanstalt, Braunschweig, Germany 4 4Center of Smart Interfaces, Technische Universit¨at Darmstadt, Darmstadt, Germany J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4452 Table 1. Selection of laboratory measurements of the accommodation coefﬁcient αice at temperatures relevant for cirrus clouds. Results are spread over almost three orders of magnitude. Previous studies are summarized in Choularton and Latham (1977), Haynes et al. (1992), and Pruppacher and Klett (1997). αice Temp. [K] Method Reference 1.06 ± 0.1 > αice > 0.65 ± 0.08 20–185 Ice layer growth Haynes et al. (1992) 0.5 ± 0.1 < αice < 1.4 ± 0.2 211–232 Ice layer Kramers and sublimation Stemerding (1951) 0.3+0.7 −0.1 200 Ice layer growth Leu (1988) 0.48 ± 0.04 > αice > 0.08 ± 0.03 140–210 Condensed ice Pratte et al. (2006) sample growth 0.031 ± 0.001 234–236 Frozen droplet Earle et al. (2010) growth 0.06 < αice < 0.07 200–219 Ice crystal growth Isono and Iwai (1969) 0.004 ± 0.002 < αice < 0.009 ± 0.003 213–233 Frozen droplet Magee et al. (2006) growth growth in cirrus clouds (in the kinetic growth regime), is the accommodation coefﬁcient of water molecules on ice. sistent supersaturations. For such high persisting supersatu- rations that have been observed in cirrus clouds, a very low ice accommodation coefﬁcient could indeed serve as expla- nation (Gao et al., 2004; Peter et al., 2006). This accommodation coefﬁcient αice, also known as the deposition coefﬁcient, is deﬁned as the sticking probability of water molecules that collide with an ice surface, e.g., of an ice particle. In the following discussion, αice will be re- ferred to as the ice accommodation coefﬁcient or simply the accommodation coefﬁcient, for brevity. The outlined potential impact on the ice particle growth and the properties of cirrus clouds make αice a relevant pa- rameter in cirrus cloud modeling. It is included not only in the formalisms for cloud ice formation in general circulation models (K¨archer et al., 2006; K¨archer and Lohmann, 2002a, b, 2003; Morrison and Gettelman, 2008) but also in more de- tailed models (Cotton et al., 2007; Spichtinger and Gierens, 2009; Haag et al., 2003). Cirrus cloud model calculations have shown that use of αice values below 0.1 going down to 0.001 can lead to a signiﬁcant increase in ice number concentration by several orders of magnitude when compared to simulations using αice = 1 (Lin et al., 2002; Gierens et al., 2003). In contrast, lowering αice from 1 to 0.1 had only little impact on the model results. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice The reason for the increase in ice number con- centration for very low αice values will be outlined in the following. However, αice is not well constrained experimentally so far, with experimental values ranging from the order of 10−3 up to unity. For a comprehensive summary of experimental results for αice from laboratory measurements, see Choular- ton and Latham (1977), Haynes et al. (1992), and Pruppacher and Klett (1997). A selection of laboratory studies that were performed at temperatures relevant for the ice growth in cir- rus clouds is given in Table 1. With respect to the applied experimental approach, they can be divided into two groups. Supersaturations with respect to ice in the upper tropo- sphere may arise by the lifting of an air parcel and the re- sulting nearly adiabatic cooling. Ice nucleation, i.e., the for- mation of an ice particle by an aerosol particle, requires that a certain supersaturation threshold is exceeded. After ice nu- cleation has set in, the growing ice particles tend to deplete the supersaturation. Hence, the reason that models predict increasing ice number concentrations with decreasing αice is that lower values of αice would lead to a higher suppres- sion of ice particle growth in cirrus clouds. This suppression in the growth would result in a higher peak supersaturation and a longer time during which the supersaturation is able to stay above the nucleation threshold, allowing more of the ice nuclei to be activated or more of the aerosols to freeze by homogeneous nucleation within the cloud. This increased ice number concentration would enhance optical depth and albedo of cirrus clouds, i.e., the radiative properties, in a sim- ilar way to the well-known Twomey effect for “warm” clouds (Lohmann and Feichter, 2005; Twomey, 1974). In addition, the ice growth suppression would lead to higher, more per- One approach observes the growth or sublimation of an ice layer or sample. Note that in the framework of ice growth used in this work (cf. Sect. 2), the sublimation coefﬁ- cient equals the ice accommodation coefﬁcient by deﬁnition. These measurements are typically carried out under pres- sures of less than 1 Pa, e.g., in an ultrahigh-vacuum chamber or a low-pressure ﬂow reactor. Under these conditions, the ice growth or sublimation rates, respectively, are directly pro- portional to αice. Th Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4453 Table 2. Overview of cirrus cloud model studies with their preferred values or ranges for the accommodation coefﬁcient αice. Studies include the examination of atmospheric cirrus clouds and of simulated cirrus clouds in cloud chamber experiments. Table 2. Overview of cirrus cloud model studies with their preferred values or ranges for the accommodation coefﬁcient αice. Studies include the examination of atmospheric cirrus clouds and of simulated cirrus clouds in cloud chamber experiments. αice Temp. [K] Study Reference 0.01 < αice < 0.1 225 Atmospheric, local Gierens et al. (2003) 0.2 preferred over 0.05 210–235 Atmospheric, local K¨archer and Str¨om (2003) > 0.1 225 Atmospheric, local Kay and Wood (2008) 0.5 preferred over 0.006 < 238 Atmospheric, global Lohmann et al. (2008) > 0.2 202 Cloud chamber Haag et al. (2003) 0.1 preferred over 1 180–200 Cloud chamber Saunders et al. (2010) are much larger and of different appearance than typical cir- rus cloud ice particles. face take place which inhibit the incorporation of water vapor molecules (Gao et al., 2004). Due to the speciﬁed relevance of the ice accommoda- tion coefﬁcient, αice, in cirrus cloud modeling, dedicated αice measurements were carried out at the aerosol and cloud chamber AIDA (Aerosol Interactions and Dynamics in the Atmosphere) by experimentally simulating the formation and evolution of cirrus ice particles under realistic conditions. These experiments are described in Sect. 3, before which we present a brief summary of the theoretical description of at- mospheric ice particle growth in Sect. 2. In order to deter- mine αice values from the AIDA experiments, two different model approaches were applied. The methods of these ap- proaches are described in Sect. 4. Combined experimental and modeling data are presented in Sect. 5. In addition, this section includes a careful uncertainty analysis in order to set appropriate lower bounds on αice. The results of our study to determine αice for cirrus ice particle growth are presented and discussed in Sect. 6. Section 7 concludes this paper. The other approach optically monitors the growth of sin- gle droplets, frozen by the homogeneous nucleation of ice, which are electrodynamically levitated in vertical wind tun- nels (Earle et al., 2010; Magee et al., 2006) or the growth of single ice crystals on a substrate (Isono and Iwai, 1969). These three experiments obtained αice values far below 0.1. The value retrieved by Earle et al. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice (2010) is αice = 0.031, the result by Isono and Iwai (1969) is 0.06 < αice < 0.07, and the results by Magee et al. (2006) suggest very low αice values in the range 0.004–0.009. g In addition to these laboratory measurements, there are several cirrus cloud model studies, summarized in Table 2, which vary αice in model calculations until good agree- ment between model output and observational data, i.e., measured ice number concentrations or ice supersaturations, is achieved. These studies have either investigated atmo- spheric cirrus cloud data from in situ airborne measurements (Gierens et al., 2003; K¨archer and Str¨om, 2003; Kay and Wood, 2008), global satellite retrievals of cloud properties (Lohmann et al., 2008) or simulated cirrus clouds in cloud chamber experiments (Haag et al., 2003; Saunders et al., 2010). All except the study by Gierens et al. (2003) are in fa- vor of an accommodation coefﬁcient greater than 0.1. Based on the results of these studies, αice is generally assumed in the range 0.2–1 for the parameterization of cirrus ice particle growth in all the different types of models mentioned pre- viously. However, αice values have either been retrieved for very limited data sets at one speciﬁc temperature (Gierens et al., 2003; Kay and Wood, 2008; Haag et al., 2003) or the magnitude of αice has not been stated to any level of precision (K¨archer and Str¨om, 2003; Lohmann et al., 2008; Saunders et al., 2010). Moreover, none of these analyses have performed a thorough uncertainty analysis for their retrieved αice values. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice Ice growth/sublimation rates are, e.g., deter- mined gravimetrically (Kramers and Stemerding, 1951) by interferometric measurement of the thickness of a plane ice layer (Haynes et al., 1992) or by measurement of the molec- ular water vapor ﬂux to the ice surface through mass spec- troscopic techniques (Leu, 1988; Pratte et al., 2006). These experiments typically yield results of αice > 0.1 for cirrus cloud temperatures. However, the ice samples investigated Atmos. Chem. Phys., 13, 4451–4466, 2013 www.atmos-chem-phys.net/13/4451/2013/ 2 Atmospheric ice particle growth Mass transfer of water molecules to the surface of atmo- spheric ice particles determines the process of their growth. The mathematical expressions for the description of this mass transfer are outlined in this section. Three cases or regimes for ice particle growth have to be distinguished dependent on the (volume equivalent) particle radius, rp, in relation to the mean free path of water vapor molecules in air λw. It is useful to deﬁne the Knudsen number Kn in this context: Kn = λw rp . (1) Kn = λw rp . (1) The Knudsen number is used to distinguish between the dif- ferent regimes of ice particle growth. If Kn ≫1, mass trans- port is determined by elementary gas kinetic processes in the so-called kinetic regime. In this regime the accommodation coefﬁcient αice plays a dominant role. On the other hand, if Kn ≪1, the ﬂux of water molecules to the ice particle is governed by diffusion in the so-called continuum regime. The determinant quantity in this regime is the diffusivity of The Knudsen number is used to distinguish between the dif- ferent regimes of ice particle growth. If Kn ≫1, mass trans- port is determined by elementary gas kinetic processes in the so-called kinetic regime. In this regime the accommodation coefﬁcient αice plays a dominant role. On the other hand, if Kn ≪1, the ﬂux of water molecules to the ice particle is governed by diffusion in the so-called continuum regime. The determinant quantity in this regime is the diffusivity of The reasons for the wide spread of values for the ice ac- commodation coefﬁcient obtained by different experimental measurements and model studies remains unknown. Some rather speculative explanations have been brought forward such as that αice could depend on particle size (Gierens et al., 2003; Magee et al., 2006) or supersaturation (Nelson and Baker, 1996) or that reactions on the ice particle sur- J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 13–76), the ice particle mass growth rate is given by dmp dt = 4πC (Sice −1) RTg ˆeice(Tg)D∗wMw + LH , (6) (6) where mp is the ice particle mass, C the electrostatic capac- itance of the ice particle divided by 4πϵ0 and referred to as the capacitance, and LH describes the effect of latent heat release from deposition of water molecules on the surface of the ice particle. However, the impact of LH is rather low for Tg < 235 K, i.e., for the growth of ice particles in cir- rus clouds. LH affects the result of Eq. (6) by around 5 % at Tg = 235 K and this effect decreases with decreasing temper- ature. Therefore, LH will not be described in detail here, but in Appendix A. The intermediate regime between kinetic and continuum regime is called transition regime (Kn ≈1) and connects the mass transfer formulation of both limiting cases. For at- mospheric conditions relevant for cirrus clouds, λw typically takes values of 200 nm and above. This means that ice parti- cles in natural cirrus clouds stay in the kinetic and interme- diate regime until they are a few micrometers in size. In order to connect the kinetic regime with the contin- uum regime, one can apply the so-called ﬂux-matching ap- proach for the transition regime (Pruppacher and Klett, 1997, Chap. 13). This approach assumes that for distances away from the ice particle surface smaller than the vapor jump length, 1v, which is typically chosen to be of the order of λw, water vapor transport is governed by elementary gas kinetic mechanisms, i.e., the kinetic regime applies. For distances greater than 1v, on the other hand, water vapor transport is governed by diffusion and the continuum regime is valid. At the boundary deﬁned by r = rp + 1v, the water vapor ﬂuxes of both regimes have to be matched. The ice particle capacitance, C, depends on the ice par- ticle shape. For spherical ice particles it is simply given by the particle’s radius rp. More complex expressions for the capacitance of spheroid ice particles are given in McDonald (1963), and good approximations for cylinders and hexag- onal columns have been determined numerically by West- brook et al. (2008). In general, the consideration of nonspher- ical ice particle shapes is of importance for high aspect ratios. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice For compact ice particles with aspect ratios up to approxi- mately two, the assumption of spherical ice particles causes an error in ice particle capacitance of ∼10 % at most. With this approach, ice particle growth within the transi- tion regime is described by a modiﬁed version of the water vapor diffusivity Dw from Eq. (2), so called D∗ w. D∗ w is given by (Pruppacher and Klett, 1997; Eqs. 13–14) www.atmos-chem-phys.net/13/4451/2013/ Atmos. Chem. Phys., 13, 4451–4466, 2013 4454 J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice water molecules in air, Dw, given by (Hall and Pruppacher, 1976) water molecules in air, Dw, given by (Hall and Pruppacher, 1976) Together with D∗ w the mass growth rate of an ice particle mainly depends on the water vapor saturation ratio with re- spect to ice, Sice. It is deﬁned by Dw = Dw,0 p0 p Tg T0 γ , (2) (2) Sice = e ˆeice(Tg), (5) Sice = e ˆeice(Tg), (5) where p is the gas pressure, Tg the gas temperature, and Dw,0 the diffusivity at p0 = 1013.25 hPa and T0 = 273.15 K. The temperature coefﬁcient γ and Dw,0 were estimated by Hall and Pruppacher (1976) for the temperature range 233–313 K to 1.94 and 0.211 cm2 s−1, respectively. We used the quite common empirical parameterization from Hall and Prup- pacher (1976) since it results in only about 8 % lower dif- fusivity values compared to a more recent analysis of ex- perimental data by Massman (1998). Please note that values calculated with a Lennard–Jones model are 20–30 % lower (Ghosh et al., 2007), which would lead to even higher ac- commodation coefﬁcients if applied in our analysis. where p is the gas pressure, Tg the gas temperature, and Dw,0 the diffusivity at p0 = 1013.25 hPa and T0 = 273.15 K. The temperature coefﬁcient γ and Dw,0 were estimated by Hall and Pruppacher (1976) for the temperature range 233–313 K to 1.94 and 0.211 cm2 s−1, respectively. We used the quite common empirical parameterization from Hall and Prup- pacher (1976) since it results in only about 8 % lower dif- fusivity values compared to a more recent analysis of ex- perimental data by Massman (1998). Please note that values calculated with a Lennard–Jones model are 20–30 % lower (Ghosh et al., 2007), which would lead to even higher ac- commodation coefﬁcients if applied in our analysis. where e is the water vapor partial pressure far away from the ice particle and ˆeice(Tg) the saturation vapor pressure with re- spect to ice as function of the gas temperature Tg. According to Pruppacher and Klett (1997; Eqs. 13–76), the ice particle mass growth rate is given by where e is the water vapor partial pressure far away from the ice particle and ˆeice(Tg) the saturation vapor pressure with re- spect to ice as function of the gas temperature Tg. According to Pruppacher and Klett (1997; Eqs. 3.1.2 Ice number concentration An optical particle counter (OPC; PALAS, WELAS) is avail- able to register ice particle number concentrations, Cn,ice, for particles in the size range 0.7–40 µm. This range has been determined for spherical particles with a refractive index of 1.33. The WELAS instrument counts particles by measuring the pulses of white light scattered by individual particles. The instrument is operated at 5 s time resolution. Its accuracy is estimated to be ±20 % (M¨ohler et al., 2006). A detailed description of the AIDA instrumentation is given in Wagner et al. (2009). The instrumentation relevant to the cirrus ice growth experiments presented in this paper is depicted in Fig. 1 and consists of the following: 3.1.3 In situ laser light scattering For the in situ measurement of water vapor concentration and partial pressure, respectively, as well as extractive measure- ment of total water content, two tunable diode laser (TDL) hygrometers – APicT and APeT – operating at a wavelength of 1370 nm are available (Skrotzki, 2012; Skrotzki et al., 2013; Ebert et al., 2005; Fahey et al., 2009). The time res- olution of these TDL hygrometers is approximately 1 s and accuracy is given at ±5 %. The in situ water vapor measure- ment is performed by APicT and the total water content is retrieved by extractive sampling of AIDA gas via a heated stainless steel line to which APeT is connected. From the difference of total water and water vapor measurements, the ice water content IWC within AIDA can be derived. The wa- ter partial pressure, e, obtained by the TDL instruments can be converted into an ice saturation ratio Sice by calculating the water vapor saturation pressure, ˆeice, with respect to the AIDA gas temperature, Tg (Murphy and Koop, 2005), ac- The in situ light scattering and depolarization instrument SI- MONE detects light scattered by aerosol or cloud particles in forward (2◦) and backward (178◦) direction. It uses a linearly polarized continuous wave semiconductor laser at 488 nm wavelength. In addition, the parallel and perpendicular po- larization components of the backscattered intensity can be detected. Due to its high sensitivity, SIMONE is used to pre- cisely determine the onset of cloud ice particle generation, i.e., the onset of ice nucleation. Further details about the in- strument can be found in Schnaiter et al. (2012). J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice APicT SIMONE WELAS OPC CPC 3010 Graphite spark generator SMPS/APS AIDA cloud simulation chamber APeT Dry powder disperser Figure 1. Schematic drawing of the AIDA cloud chamber with instrume measurements (APicT and APeT), ice particle characterization (SIM optical particle counter), aerosol generation (graphite spark generato disperser), and aerosol characterization (CPC 3010, SMPS, and APS illustrates the thermostated insulating housing surrounding the AIDA cham Fig. 1. Schematic drawing of the AIDA cloud chamber with in- strumentation for humidity measurements (APicT and APeT), ice particle characterization (SIMONE and WELAS optical particle counter), aerosol generation (graphite spark generator and dry- powder disperser), and aerosol characterization (CPC 3010, SMPS, and APS). The gray frame illustrates the thermostated insulating housing surrounding the AIDA chamber APicT SIMONE WELAS OPC CPC 3010 Graphite spark generator SMPS/APS AIDA cloud simulation chamber APeT Dry powder disperser conditions are characteristic of cirrus clouds formed by oro- graphic waves (Field et al., 2001; Gayet et al., 2006) and re- sulted in realistic cirrus ice particle growth yielding repre- sentative particle sizes and shapes (cf. the discussion on ice particle shapes in Sect. 4). conditions are characteristic of cirrus clouds formed by oro- graphic waves (Field et al., 2001; Gayet et al., 2006) and re- sulted in realistic cirrus ice particle growth yielding repre- sentative particle sizes and shapes (cf. the discussion on ice particle shapes in Sect. 4). In the following, an overview of the AIDA chamber and the instrumentation relevant to this work, the aerosol types utilized in the experiments, and the experimental parameters and methods will be given. 3.1 AIDA instrumentation and aerosol types The AIDA chamber consists of an aluminum vacuum vessel with a diameter of 4 m, a height of 7.5 m, and a volume of 84.3 m3. This large volume keeps boundary effects from the aluminum wall such as temperature and humidity gradients conﬁned to a small fraction of the total volume. The vessel is placed in an isolating and thermostated housing which allows an operation temperature range between −90 ◦C and +60 ◦C. The gas temperature inside the AIDA chamber is measured to an accuracy of ±0.3 K (M¨ohler et al., 2006). A mixing fan maintains homogeneous conditions in the gas volume inside the vessel, which results in temperature differences of less than ±0.2 K within the entire gas volume under static con- ditions (M¨ohler et al., 2006). Two vacuum pumps allow gas pressures from ambient pressure down to 0.01 hPa. Available cooling rates range from 0.1 K min−1 to 6 K min−1 and are a result of nearly adiabatic cooling by gas pressure reduction due to pumping. This cooling process is used in AIDA ex- periments to simulate the quasi-adiabatic expansion cooling that ascending air parcels experience in the atmosphere. Figure 1. Schematic drawing of the AIDA cloud chamber with instrume measurements (APicT and APeT), ice particle characterization (SIM optical particle counter), aerosol generation (graphite spark generat disperser), and aerosol characterization (CPC 3010, SMPS, and AP illustrates the thermostated insulating housing surrounding the AIDA cha Fig. 1. Schematic drawing of the AIDA cloud chamber with in- strumentation for humidity measurements (APicT and APeT), ice particle characterization (SIMONE and WELAS optical particle counter), aerosol generation (graphite spark generator and dry- powder disperser), and aerosol characterization (CPC 3010, SMPS, and APS). The gray frame illustrates the thermostated insulating housing surrounding the AIDA chamber cording to Eq. (5). The accuracy of the retrieval of Sice is therefore not only determined by the accuracy of the TDL instruments but also by the uncertainty of the gas tempera- ture Tg. 3 Experimental methods For the determination of the ice accommodation coefﬁcient αice, dedicated experiments at the cloud chamber AIDA (M¨ohler et al., 2003, 2006) with simulated cirrus clouds in the temperature range between 190 K and 235 K have been performed. The ice particles were created by deposition nu- cleation (M¨ohler et al., 2006) on synthetic hematite aerosol particles as well as graphite spark generator (GSG) soot. The utilization of deposition nucleation allowed very small initial sizes of the ice particles, below 100 nm, which resulted in the experiments having a high sensitivity to αice. Cooling rates during the dynamic expansion experiments were between 0.5 K min−1 and 2.7 K min−1, and experimental peak super- saturations varied between moderate supersaturations and su- persaturations close to the homogeneous freezing threshold of supercooled solution droplets (Koop et al., 2000). These D∗ w = Dw rp rp+1v + 4Dw αicerp ¯cw , (3) (3) where ¯cw is the mean thermal speed of water vapor molecules. It is given by (Pruppacher and Klett, 1997; Eqs. 5–49) ¯cw = 8RTg πMw 1/2 , (4) ¯cw = 8RTg πMw 1/2 , (4) with the universal gas constant R and the molar mass of wa- ter Mw. From Eq. (3), one obtains the limiting cases of the continuum as well as the kinetic regime for the correspond- ing limits of rp. In the limit rp →∞, D∗ w →Dw (contin- uum regime). In the limit rp →0, D∗ w →αicerp ¯cw/4 (kinetic regime), where αice plays a dominant role. www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ Atmos. Chem. Phys., 13, 4451–4466, 2013 Atmos. Chem. Phys., 13, 4451–4466, 2013 4455 3.1.4 Aerosol generation For injection of aerosol particles into the AIDA chamber, the following aerosol generators have been used: a dry-powder disperser (TSI, model 3433) for the addition of two different www.atmos-chem-phys.net/13/4451/2013/ 3.1.5 Aerosol characterization The aerosol number concentration, Cn,ae, is measured by a condensation particle counter (TSI, CPC 3010) at a time res- olution of 1 s. Its accuracy is estimated to be ±20 % . Aerosol size distributions are determined by a scanning mobility par- ticle sizer (SMPS, TSI) in combination with an aerodynamic particle sizer (APS, TSI). From these measurements, the me- dian aerosol size µae and the width parameter σae of a lognor- mal ﬁt to the measured aerosol distribution are estimated to be retrieved with an accuracy of ±10 % and ±15 %, respec- tively; see Sect. 3.2. Aerosol size distribution measurements were carried out close before the start of each AIDA experi- ment. Aerosol surface size distributions for two of the AIDA ex- periments listed in Table 3 are given in Fig. 2. They are ob- tained from SMPS number size distribution measurements. For the hematite particles, the size distributions are given with respect to their volume-equivalent diameter. A dynamic shape factor of 1.1 was assumed for hematite #1 and 1.0 for hematite #2. For GSG soot the size distributions are given with respect to the electrical mobility diameter. Lognormal ﬁts were applied to the experimental aerosol surface size distributions, bimodal for the hematite aerosol consisting of monomers, as well as agglomerates and monomodal for GSG soot. The lognormal functions are described by the median aerosol size µae and the width parameter σae. The size distri- butions show that aerosol particles greater than 1 µm are neg- ligible for both aerosol types and conﬁrm the mean aerosol sizes given above. Rigorous cleaning by evacuating the AIDA vessel to pressures below 0.1 hPa and purging with synthetic air when changing the aerosol type resulted in very low back- ground concentrations of aerosol particles (typically below 0.1 cm−1) before addition of the aerosol. Hematite and GSG soot were used as aerosol particles due to their efﬁciency as ice nuclei in the temperature range 190–235 K (Gallavardin et al., 2008; M¨ohler et al., 2005). A second reason for the choice of hematite particles and GSG soot as aerosol was that these aerosol particles are hardly detected by the WELAS OPC due to their small size and low reﬂectivity, which re- duces interference of aerosol particle signals in the ice num- ber concentration measurement to a minimum. 3.1.5 Aerosol characterization This interfer- ence is caused by aerosol particles larger in optical diameter than the lower detection limit of the WELAS OPC of 0.7 µm. Of the two hematite particle samples used for the exper- iments, sample one (hematite #1) consists of nearly spheri- cal particles with a mean diameter of approximately 200 nm, while hematite sample two (hematite #2) consists of prolate spheroids with an aspect ratio of nearly two with mean ma- jor extension of approximately 500 nm. These characteris- tics of the hematite particles have been determined by scan- ning electron microscopy (Vragel, 2009). The GSG soot par- ticles have sizes mainly in the range 100–200 nm as de- scribed above. For the size distribution of the aerosol types, an aerosol background in the WELAS OPC measurement of below 1 cm−3 was obtained for GSG soot and hematite #1 as well as below 10 cm−3 for hematite #2. In order to subtract this aerosol background from the OPC ice number concentra- tion measurement, a lower cut-off at 1 µm particle size was applied to the OPC data for all AIDA experiments. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ic 4456 3.2 Overview of experiments samples of synthetic hematite particles and a graphite spark generator (PALAS, GFG 1000) which creates soot particles by spark discharge between two electrodes of pure carbon. The obtained GSG soot particles have sizes mainly in the range 100–200 nm and are agglomerates of individual soot particles with diameters below 10 nm. For details on cre- ation, morphology, and properties of GSG soot, see M¨ohler et al. (2005) and references therein. Table 3 gives an overview of the AIDA cirrus cloud exper- iments carried out for the determination of the ice accom- modation coefﬁcient αice. As mentioned above, they spread a temperature range of approximately 190–235 K to indi- cate a potential dependency of αice on temperature. A wide variety of maximum ice number concentrations Cn,ice, be- tween approximately 40 cm−3 and 200 cm−3, and maximum ice saturation ratios Sice, from moderate supersaturations of 16 % up to supersaturations close to the homogeneous freez- ing threshold of supercooled solution droplets (Koop et al., 2000), were achieved. This variety in Cn,ice and Sice was deemed appropriate to demonstrate a potential dependence of αice on ice particle size or supersaturation. Conducting the experiments at lower pressures would demonstrate higher sensitivity for the accommodation coefﬁcient of water on the growing ice crystals; however, here we started the expan- sion experiments at approximately 1 atm. Test experiments at 200 hPa gave lower cooling rates and therefore supersat- urations and ice crystal number concentrations were smaller compared to 1 atm. These difﬁculties compensated the poten- tial gain expected for operating at lower pressures. www.atmos-chem-phys.net/13/4451/2013/ Atmos. Chem. Phys., 13, 4451–4466, 2013 J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4457 HALO05_24 HALO06_26 1 2 3 Figure 2. Aerosol surface size distributions for the experiments HALO05_24 with hemat 4 Fig. 2. Aerosol surface size distributions for the experiments HALO05 24 with hematite #1 aerosol and HALO06 26 with GSG soot aerosol. The surface size distributions are inferred from number size distributions as measured by a scanning mobility particle sizer SMPS. Lognormal ﬁts (solid lines) are applied to the measurement data (open symbols), bimodal for the hematite aerosol (monomers and agglomerates), and monomodal for the graphite spark generator soot aerosol. HALO05_24 HALO06_26 1 2 3 Figure 2. Aerosol surface size distributions for the experiments HALO05_24 with hema 4 Fig. 2. Aerosol surface size distributions for the experiments HALO05 24 with hematite #1 aerosol and HALO06 26 with GSG soot aerosol. The surface size distributions are inferred from number size distributions as measured by a scanning mobility particle sizer SMPS. Lognormal ﬁts (solid lines) are applied to the measurement data (open symbols), bimodal for the hematite aerosol (monomers and agglomerates), and monomodal for the graphite spark generator soot aerosol. aerosol and HALO06_26 with GSG soot aerosol. The surface size distributions are in 5 from number size distributions as measured by a scanning mobility particle sizer S 6 Lognormal fits (solid lines) are applied to the measurement data (open symbols), bimod 7 Table 3. Overview of AIDA accommodation coefﬁcient experiments sorted by aerosol type and temperature. Two types of aerosols were used: hematite particles (two different samples) and graphite spark generator (GSG) soot. p(t0) and Tg(t0) indicate gas pressure and temperature at start time t0 of the experiments. The experiments cover a broad temperature range relevant for cirrus clouds (approximately between 190 K and 235 K) and a wide variety of initial aerosol number concentrations Cn,ae(t0), maximum ice number concentrations Cn,ice, and maximally obtained ice saturation ratios Sice. aerosol and HALO06_26 with GSG soot aerosol. The surface size distributions are in 5 from number size distributions as measured by a scanning mobility particle sizer S 6 Lognormal fits (solid lines) are applied to the measurement data (open symbols), bimod 7 Table 3. Overview of AIDA accommodation coefﬁcient experiments sorted by aerosol type and temperature. Two types of aerosols were used: hematite particles (two different samples) and graphite spark generator (GSG) soot. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice p(t0) and Tg(t0) indicate gas pressure and temperature at start time t0 of the experiments. The experiments cover a broad temperature range relevant for cirrus clouds (approximately between 190 K and 235 K) and a wide variety of initial aerosol number concentrations Cn,ae(t0), maximum ice number concentrations Cn,ice, and maximally obtained ice saturation ratios Sice. the hematite aerosol (monomers and agglomerates) and monomodal for the grap 8 generator soot aerosol. 9 10 Exp. no. Aerosol particles p(t0) [hPa] Tg (t0) [K] Cn,ae(t0) [cm−3] max[Cn,ice] [cm−3] max[Sice] HALO06 19 Hematite #1 1008.5 234.9 315 111 1.23 HALO06 20 Hematite #1 1011.7 234.9 192 88 1.31 HALO06 21 Hematite #1 1011.3 225.0 287 72 1.16 HALO06 22 Hematite #1 1011.9 224.5 183 61 1.24 HALO05 18 Hematite #1 1009.1 213.7 189 90 1.26 HALO04 05 Hematite #2 995.4 212.5 280 63 1.36 HALO05 24 Hematite #1 1005.7 198.1 185 60 1.69 HALO04 09 Hematite #2 965.9 196.4 –∗ 56 1.51 HALO06 23 GSG soot 1015.3 233.9 1976 153 1.30 HALO06 24 GSG soot 1015.9 234.0 862 121 1.33 HALO06 25 GSG soot 1015.5 224.3 321 72 1.27 HALO06 26 GSG soot 1015.3 223.7 164 65 1.30 HALO06 27 GSG soot 1019.7 212.8 269 80 1.45 HALO06 28 GSG soot 1019.9 213.0 145 37 1.41 HALO04 26 GSG soot 1011.4 198.2 2410 197 1.37 ∗no measurement available ∗no measurement available 2011; Earle et al., 2010) for temperatures below −35 ◦C. The maximum size to which the ice particles in all experiments listed in Table 3 grew stayed below 20 µm. where ρice is the mass density of ice. The assumption of spherical ice particles also implies that no exposed facets of enhanced growth (Libbrecht, 2005) exist. Furthermore, the analysis of SIMONE depolarization ratio data for HALO06 25 and HALO06 27 as well as two AIDA ice nucleation experiments with GSG soot at approximately 200 K shows that prolate spheroids with a maximum aspect ratio of two represent the entire ensemble of ice particles The assumption of spherical ice particles is justiﬁed since ice particles smaller than 20 µm in diameter have been ob- served to be compact and nearly spherical in cirrus cloud measurements (Korolev and Isaac, 2003; Mitchell et al., 2011) as well as in laboratory studies (Abdelmonem et al., Atmos. Chem. Phys., 13, 4451–4466, 2013 4 Modeling methods In order to retrieve values for the ice accommodation co- efﬁcient αice from the AIDA cirrus cloud experiments, ex- perimental data were evaluated with two independent model approaches. The ﬁrst model used is SIGMA (Simple Ice Growth Model for determining Alpha), the second one is ACPIM (Aerosol-Cloud-Precipitation Interaction Model). Both models apply Eq. (6) for the parameterization of cir- rus ice particle growth and assumed spherical ice particle shape. This implies that ice particle size and mass are con- nected by rp = 3 4π mp ρice 1/3 , (7) (7) Atmos. Chem. Phys., 13, 4451–4466, 2013 www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ 4.1 SIGMA The ice growth model SIGMA has been developed as a ded- icated tool to model the growth of cirrus cloud ice particles dependent on the magnitude of the accommodation coefﬁ- cient αice. A list of all physical quantities used in SIGMA is given in Appendix B. 6 7 8 yields the ice growth start times it in the subsequent ic The time-dependent total ice number concentration Cn,ice, the gas pressure p, and the gas temperature Tg are SIGMA input parameters along with the ice saturation ratio Sice of water vapor which is required in Eq. (6). These measured quantities are linearly interpolated to match the model time resolution 1t. SIGMA relies on the Dahneke approach (Dahneke, 1983) for a choice of the vapor jump length 1v in Eq. (3). This approach sets 9 1v = λw, (8) (8) 1v = λw, For the application of SIGMA to the AIDA experiments listed in Table 3, the calculation of IWCSIGMA covered time spans approximately between 100 s and 500 s with a time res- olution 1t = 0.1 s. Ice particle growth was segmented into N = 20 individual size bins with the initial diameter of the ice particles in each ice growth bin set to 1 µm, which corre- sponds to the cut-off set to safely separate between the WE- LAS OPC counts for aerosol and ice particles. where the mean free path λw is connected to the diffusivity Dw by where the mean free path λw is connected to the diffusivity Dw by λw = 2Dw ¯cw . (9) λw = 2Dw ¯cw . (9) As output quantity, SIGMA calculates the time-dependent total ice water content IWCSIGMA inside AIDA for a seg- mentation of ice particle growth into individual bins. Each individual bin is indexed by i and has a different start time of ice growth ti. This is illustrated in Fig. 3. Of a total number of ice growth bins N, a fraction n(t) is active at time t. Each bin contains ice particles of mass mp[i] with ice number con- centration Cn,ice[i]. By summing over all active bins SIGMA calculates IWCSIGMA in terms of volume mixing ratio by the following relation: J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4458 present in these experiments well, cf. Schnaiter et al. (2012). These results refer to a temperature range approximately be- tween 200 K and 225 K and can be used to estimate an upper bound for the error resulting from the assumption of spheri- cal particles. Spheroids with an aspect ratio of two result in a deviation of 4 % in ice particle capacitance Cs (McDonald, 1963) compared to the capacitance rp of a volume-equivalent sphere. The ice particle shape cannot be excluded as being hexagonal columnar for a fraction of the ice particles in the entire ensemble, but even in this case the capacitance Ch (Westbrook et al., 2008; Westbrook and Heymsﬁeld, 2011) for an aspect ratio of two would not deviate more than 11 % from rp of a volume-equivalent sphere. Therefore, it is ex- pected that the assumption of spherical ice particles does not have signiﬁcant impact on the retrieval of the accommoda- tion coefﬁcient αice. 1 2 0 100 200 300 400 500 700 800 900 1000 190 195 200 p [hPa] Relative time [s] T [K] 0 10 20 30 40 50 60 Cn,ice [cm -3] Figure 3. Division of ice particle growth into individua a typical evolution of the ice number concentratio experiment as measured by the WELAS optical particl dashed dotted line and is chosen as start time 1t for Fig. 3. Division of ice particle growth into individual bins in SIGMA. The blue line shows a typical evolution of the ice num- ber concentration Cn,ice during an AIDA expansion experiment as measured by the WELAS optical particle counter. Ice onset is indi- cated by the dashed dotted line and is chosen as start time t1 for ice growth in the ﬁrst ice growth bin. Equidistant division of Cn,ice with respect to the maximally reached ice number concentration yields the ice growth start times ti in the subsequent ice growth bins (dot- ted lines). Model speciﬁc details about SIGMA and APCIM and their application to the AIDA cirrus cloud experiments are now given. 3 4 www.atmos-chem-phys.net/13/4451/2013/ t al.: Accommodation coefﬁcient of water molecules on ice 0 100 200 300 400 500 700 800 900 1000 190 195 200 p [hPa] Relative time [s] T [K] 0 10 20 30 40 50 60 Cn,ice [cm -3] Figure 3. Division of ice particle growth into individua a typical evolution of the ice number concentratio experiment as measured by the WELAS optical particl dashed dotted line and is chosen as start time 1t for Fig. 3. Division of ice particle growth into individual bins in SIGMA. The blue line shows a typical evolution of the ice num- ber concentration Cn,ice during an AIDA expansion experiment as measured by the WELAS optical particle counter. Ice onset is indi- cated by the dashed dotted line and is chosen as start time t1 for ice growth in the ﬁrst ice growth bin. Equidistant division of Cn,ice with respect to the maximally reached ice number concentration yields the ice growth start times ti in the subsequent ice growth bins (dot- ted lines). 5 Experimental and modeling data The probability density functions used for the variation are, in the case of SIGMA, normal distributions with a stan- dard deviation corresponding to the respective measurement accuracy. In the case of ACPIM, these functions are given by even probability distributions within the interval deﬁned by the measurement accuracy, which gives a more conservative error bar on the ﬁnal output. The accuracies taken as basis for the uncertainty analysis are given in Table 4 and are based on the measurement accuracies discussed in Sect. 3.1. As Sice and IWC are not retrieved from independent measurements, the variation of these two quantities is coupled. For ACPIM, it does not make sense to adjust the total water measurement by its accuracy since conditions were close to ice saturation at the start of the experiments. Combined measurement and model data based on the de- scription of Sects. 3 and 4 for the AIDA experiments HALO05 24 with hematite aerosol and HALO06 26 with GSG soot aerosol are presented in Fig. 4. Experimental data of the ice water content IWC are compared with the SIGMA modeled ice water content IWCSIGMA for different values of the accommodation coefﬁcient αice. Correspondingly, inde- pendent ACPIM calculations of the temporal evolution of ice saturation ratio Sice for different values of αice are presented together with the according measurement data. For both ex- periments, SIGMA as well as ACPIM suggest αice > 0.1 and are in good agreement with respect to each other despite their different approaches to retrieve αice. p In the case of SIGMA, the Monte Carlo simulation con- sists of 1000 model runs for each AIDA experiment. For the randomly varied data sets of each run, the SIGMA model obtains a best-ﬁt value α∗ ice by comparing the model output IWCSIGMA to the experimentally determined IWC. This is done by means of a Levenberg–Marquardt ﬁtting algorithm (Marquardt, 1963; Press et al., 2007) which varies αice un- til a best-ﬁt value α∗ ice resulting in optimal agreement be- tween IWCSIGMA and experimental IWC is obtained. The ﬁt- ting algorithm is constrained to α∗ ice ≤1 in this procedure as α∗ ice > 1 does not represent a physically plausible state. The resulting distribution of 1000 α∗ ice values allows computing of an overall best-ﬁt value with uncertainty bounds for each AIDA experiment. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice on this best-ﬁt value is obtained for each AIDA experiment. This is done by means of an uncertainty analysis based on the Monte Carlo method (Rubinstein and Kroese, 2008). while the humidity is allowed to vary and depends on the growth of the ice by vapor deposition. In each model time step, the rate of change of temperature was speciﬁed so that it equaled the measured value during the experiment. The ice crystals are assumed to grow from the aerosol size distribution as ice is nucleated on the aerosol particles and form at a rate that was measured using the WE- LAS OPC. An assumption was that ice nucleation occurred in proportion to the surface area of the aerosol particles so that the largest particles have the highest chance of nucleat- ing ice; this was found to be the case in other similar exper- iments (Saunders et al., 2010). Once an aerosol particle has nucleated ice it is no longer available to nucleate further ice. The ice particles then grow by vapor deposition, depleting the available water vapor; however, an additional term is also added to the modeled humidity for each time step so that the total water content in the model equals that which was mea- sured throughout the experiment. A comparison of the mod- eled and measured humidity then allows for an assessment of the level of agreement between model and data. The Monte Carlo method was implemented in the follow- ing manner, cf. Cullen and Frey (1999). For each run of the Monte Carlo simulation, SIGMA and ACPIM input data sets as well as the experimentally determined data set to which the model output is compared are randomly varied accord- ing to their accuracy. Note that, in general, the measurement precision is of minor importance for the measurement un- certainty and therefore not taken into account. Uncertainties of gas pressure p with ±1 hPa and gas temperature Tg with ±0.3 K (M¨ohler et al., 2006) are expected to be insigniﬁcant and are neglected as well. Although uncertainties in thermophysical quantities like diffusion coefﬁcients (see Sect. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 2) or latent heat (see Ap- pendix A) contribute to the overall uncertainty for the accom- modation coefﬁcients (for example in the experimental con- ﬁguration of Miles et al., 2012), we refrain from including them in this systematic uncertainty analysis since the dom- inating factors for our work are uncertainties in the experi- mental observables. Uncertainty analysis ice In the case of ACPIM, simulations for 13 of the 15 ex- periments given in Table 3 were considered, and for each of them 100 model runs with different values of the input From the SIGMA and ACPIM model calculations, a best-ﬁt value for the accommodation coefﬁcient with an uncertainty 5 Experimental and modeling data The overall best-ﬁt value is given by the median, its lower bound by the lower quartile, and its upper bound by the upper quartile of the α∗ ice distribution. Note that here and for all other experiments listed in Ta- ble 3, a constant offset correction was applied to the experi- mental data set of APeT total water content in order to match it with the APicT water vapor content at ice onset of each ex- periment. This approach assumes that the IWC inside AIDA is zero until the onset of ice nucleation. The necessity of this correction by up to 7 % for Tg < 200 K may be because of the APeT extractive sampling at one point inside the AIDA chamber close to the vessel wall, which may be inﬂuenced by slight gas heterogeneities inside AIDA during the dynamic expansions. This could cause deviations when compared to the APicT water vapor measurement, which is more repre- sentative of humidity conditions along the entire diameter of the chamber. 4.2 ACPIM The Aerosol-Cloud-Precipitation Interaction Model has been described and used for nucleation studies in the AIDA by Connolly et al. (2009). More thorough and up-to-date de- scriptions of the numerical methods used are provided by Dearden et al. (2011) and Connolly et al. (2012). Essentially, it is run as a bin-microphysical parcel model in which the aerosol size distribution is discretized over a bin grid and the growth of ice particles and interaction with the temperature and water vapor ﬁeld are solved as a coupled set of ordi- nary differential equations. In this paper only the process of growth of ice by vapor deposition was considered. ACPIM was constrained to the measurements of temperature, pres- sure, total water, and ice particle number concentration, IWCSIGMA= RTg Mwp n(t)≤N X i=1 Cn,ice[i]mp[i], (10) (10) where mp[i] is obtained by the integration of Eq. (6) mp[i] = t ∫ ti dmp dt 1t. (11) (11) Atmos. Chem. Phys., 13, 4451–4466, 2013 Atmos. Chem. Phys., 13, 4451–4466, 2013 Atmos. Chem. Phys., 13, 4451–4466, 2013 Atmos. Chem. Phys., 13, 4451–4466, 2013 www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ 4459 J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice The dotted line indicates the start time of the experiment, i.e., the start of pumping, while the dashed dotted line depicts the ice onset time inferred from SIMONE forward scattering data. vapor content, ice water content IWC inferred measurement, ice number concentration n,ice C Table 4. Accuracies of the experimental data sets used for the SIGMA and ACPIM model uncertainty analysis based on the Monte Carlo method. The points mark if the data set is randomly varied in the uncertainty analysis of the respective model. saturation ratio ice S derived from APicT wat 7 from APicT water vapor and APeT total wat 8 measured by the WELAS optical particle co 9 gT , and AIDA wall temperature w T . For a r 10 from the SIGMA model SIGMA IWC for th 11 ACPIM model for the evolution of ice S are i 12 i di t th t t ti f th i t i 13 parameters were performed with seven different values of the ice accommodation coefﬁcient: αice = 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, and 1.0. This made a grand total of 9100 ACPIM simulations. For each run we then calculated the sum of the squares of the residual between the measured humidity and the model for 600 s of the experiment (see Fig. 5a). In order to ensure that the Monte Carlo simulation was unbiased, we generated a vector of values of αice on an equidistant spaced grid, and the residuals at values of αice that were not modeled were estimated by linear interpolation (which was reasonable in this case as there was a smooth variation of the sum of squares with αice). This gives a large distribution of the sum of squares (e.g., Fig. 5b). er as well as gas pressure p , gas temperature ge of accommodation coefficients ice  , results volution of ice water content and from the uded in the respective panels. The dotted line Accuracy SIGMA ACPIM Sice water vapor ±5 % • Ice water content IWC ±5 % • Ice number concentration Cn,ice ±20 % • • Aerosol number concentration Cn,ae ±20 % • Median aerosol size µae ±10 % • Aerosol width parameter σae ±15 % • indicates the start time of the experiment, i. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 1 2 Figure 4. Experimental measurements and model calculations for one experiment with 3 hematite aerosol at low temperatures (HALO05_18) and one experiment with graphite spark 4 generator soot aerosol at intermediate temperatures (HALO06_26). The aerosol surface size 5 Fig. 4. Experimental measurements and model calculations for one experiment with hematite aerosol at low temperatures (HALO05 18) and one experiment with graphite spark generator soot aerosol at intermediate temperatures (HALO06 26). The aerosol surface size distributions for these experiments are given in Fig. 2. Panels from top to bottom show ice saturation ratio Sice derived from APicT water vapor content, ice water content IWC inferred from APicT water vapor and APeT total water measurement, ice number concentration Cn,ice measured by the WELAS optical particle counter as well as gas pressure p, gas temperature Tg, and AIDA wall temperature Tw. For a range of accommodation coefﬁcients αice, results from the SIGMA model IWCSIGMA for the evolution of ice water content and from the ACPIM model for the evolution of Sice are included in the respective panels. The dotted line indicates the start time of the experiment, i.e., the start of pumping, while the dashed dotted line depicts the ice onset time inferred from SIMONE forward scattering data. 2 Figure 4. Experimental measurements and model calculations for one experiment with 3 hematite aerosol at low temperatures (HALO05_18) and one experiment with graphite spark 4 generator soot aerosol at intermediate temperatures (HALO06_26). The aerosol surface size 5 Fig. 4. Experimental measurements and model calculations for one experiment with hematite aerosol at low temperatures (HALO05 18) and one experiment with graphite spark generator soot aerosol at intermediate temperatures (HALO06 26). The aerosol surface size distributions for these experiments are given in Fig. 2. Panels from top to bottom show ice saturation ratio Sice derived from APicT water vapor content, ice water content IWC inferred from APicT water vapor and APeT total water measurement, ice number concentration Cn,ice measured by the WELAS optical particle counter as well as gas pressure p, gas temperature Tg, and AIDA wall temperature Tw. For a range of accommodation coefﬁcients αice, results from the SIGMA model IWCSIGMA for the evolution of ice water content and from the ACPIM model for the evolution of Sice are included in the respective panels. www.atmos-chem-phys.net/13/4451/2013/ Atmos. Chem. Phys., 13, 4451–4466, 2013 4460 38 www.atmos-chem-phys.net/13/4451/2013/ J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice A Monte Carlo simulation is used to generate the cumulative fraction of residuals (c) and then a signiﬁcance level for the error bar is assigned (25 %) to ﬁnd the critical value of the residual above which the observation and model are deemed to be signiﬁcantly different. Remapping this to ﬁnd the corresponding αice gives the conﬁdence interval for alpha (d). 2 Figure 5. A schematic of the technique used to find the best guess and confidence interval of 3 the ice accommodation coefficient ice  using ACPIM. Panel (a) shows a schematic of the 4 observed and model calculated relative humidity for one choice of ice  . The sum of squares of 5 Fig. 5. A schematic of the technique used to ﬁnd the best guess and conﬁdence interval of the ice accommodation coefﬁcient αice using ACPIM. Panel (a) shows a schematic of the observed and model-calculated relative humidity for one choice of αice. The sum of squares of the residual is calculated for each 10 second time interval of the experiment from 0 to 600 s. This is done for different values of αice, so that a graph of the sum of squares vs. αice can be produced (b). A Monte Carlo simulation is used to generate the cumulative fraction of residuals (c) and then a signiﬁcance level for the error bar is assigned (25 %) to ﬁnd the critical value of the residual above which the observation and model are deemed to be signiﬁcantly different. Remapping this to ﬁnd the corresponding αice gives the conﬁdence interval for alpha (d). a graph of the sum of squares vs. ice  can be sed to generate the cumulative fraction of the error bar is assigned (25%) to find the observation and model are deemed to be he corresponding ice  gives the confidence SIGMA and ACPIM according to Sect. 5.1. The results are presented in Fig. 6. For all individual experiments, αice > 0.2 is preferred by both models and αice < 0.1 is excluded by the respective uncertainty bounds. ACPIM seems to prefer lower αice values with increasing temperature, but no sig- niﬁcant temperature dependence of αice can be observed in the ACPIM and SIGMA results. Therefore, average values of αice valid for the given temperature range are computed to be αSIGMA ice = 0.9+0.1 −0.5 and αACPIM ice = 0.5+0.5 −0.3. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice In addition, no indication for a dependence of αice on ice particle size or on supersaturation has been found for the considered ice parti- cles smaller than 20 µm and supersaturations of up to 70 %. This is done for different values of ice  , so th 7 produced (b). A Monte Carlo simulation is 8 residuals (c) and then a significance level fo 9 critical value of the residual above which 10 speciﬁed there to be poor agreement between model and data. To ﬁnd the value of αice that this corresponds to, we used the data indices to ﬁnd all of the αice values above this point; the minimum value of those αice corresponds to the lower quar- tile and the maximum to the upper quartile (e.g., Fig. 5d). To ﬁnd the median we did the same procedure except used a cumulative fraction of 0.5 (Fig. 5d). significantly different. Remapping this to fin 11 interval for alpha (d). 12 13 Note that in the case of SIGMA, the given accuracy for the measurement of water vapor Sice is the factor dominating the obtained uncertainty limits except for the experiments at lowest temperatures around 200 K for which uncertainty of the ice number concentration Cn,ice starts to become equally important. The contribution of IWC accuracy is of minor im- portance for all experiments. For ACPIM, uncertainties in aerosol size distribution parameters and ice number concen- trations tend to have the largest effect on the sum of squares of residuals. 39 ACPIM results in a somewhat lower value for αice than SIGMA, but within the uncertainty limits SIGMA and ACPIM results are in good agreement with respect to each other. Therefore, the results of both models are combined to one overall result: αice = 0.7+0.3 −0.5. A comparison of this over- all result with existing literature data is depicted in Fig. 7. The given literature values are based on laboratory measure- ments of αice (cf. Table 1) and cirrus cloud model studies (cf. Table 2). J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4461 1 1 2 Figure 5. A schematic of the technique used to find the best guess and confidence interval of 3 the ice accommodation coefficient ice  using ACPIM. Panel (a) shows a schematic of the 4 observed and model calculated relative humidity for one choice of ice  . The sum of squares of 5 Fig. 5. A schematic of the technique used to ﬁnd the best guess and conﬁdence interval of the ice accommodation coefﬁcient αice u ACPIM. Panel (a) shows a schematic of the observed and model-calculated relative humidity for one choice of αice. The sum of squ of the residual is calculated for each 10 second time interval of the experiment from 0 to 600 s. This is done for different values of so that a graph of the sum of squares vs. αice can be produced (b). A Monte Carlo simulation is used to generate the cumulative frac of residuals (c) and then a signiﬁcance level for the error bar is assigned (25 %) to ﬁnd the critical value of the residual above which observation and model are deemed to be signiﬁcantly different. Remapping this to ﬁnd the corresponding αice gives the conﬁdence int for alpha (d). 2 Figure 5. A schematic of the technique used to find the best guess and confidence interval of 3 the ice accommodation coefficient ice  using ACPIM. Panel (a) shows a schematic of the 4 observed and model calculated relative humidity for one choice of ice  . The sum of squares of 5 Fig. 5. A schematic of the technique used to ﬁnd the best guess and conﬁdence interval of the ice accommodation coefﬁcient αice using ACPIM. Panel (a) shows a schematic of the observed and model-calculated relative humidity for one choice of αice. The sum of squares of the residual is calculated for each 10 second time interval of the experiment from 0 to 600 s. This is done for different values of αice, so that a graph of the sum of squares vs. αice can be produced (b). 6 Results and discussion For the ﬁfteen AIDA cirrus cloud experiments covering a temperature range between 190 K and 235 K (see Table 3), overall best-ﬁt values of the ice accommodation coefﬁcient αice along with uncertainty bounds have been obtained by Classiﬁcation of the presented results with respect to pre- vious cirrus cloud model studies generally shows good agree- ment. A possible source of uncertainty in the model studies Atmos. Chem. Phys., 13, 4451–4466, 2013 J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 13 line depicts the ice onset time inferred from S 14 15 To ﬁnd the values of αice that gave the lowest sums of squares residuals, we took the sums of squares of residuals for each experiment and created a histogram with 30 loga- rithmically spaced bins, following which we calculated the 38 the start of pumping, while the dashed dotted MONE forward scattering data. cumulative fraction histogram of the sums of squares of the residuals (e.g., Fig. 5c). We then found the bin which had a cumulative fraction larger than 0.75, which was deﬁned as the critical value of sums of squares above which we Atmos. Chem. Phys., 13, 4451–4466, 2013 38 www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice 4462 1 2 Figure 6. Accommodation coefficients ice  obtained from SIGMA and ACPIM model 3 calculations for the AIDA experiments listed in Table 3. Best-fit values relate to the median 4 Fig. 6. Accommodation coefﬁcients αiceobtained from SIGMA and ACPIM model calculations for the AIDA experiments listed in Table 3. Best-ﬁt values relate to the median of the distributions of αice values that were retrieved by the uncertainty analysis described in Sect. 5.1, while error bars indicate the respective lower and upper quartiles. Likewise, median as well as lower and upper quartiles were determined from the temperature distribution of each experiment and applied accordingly. The temperature-averaged αice values obtained by SIGMA αSIGMA ice = 0.9+0.1 −0.5 and ACPIM αACPIM ice = 0.5+0.5 −0.3 are given by the solid black lines with the shaded region indicating their uncertainty. 1 2 Figure 6. Accommodation coefficients ice  obtained from SIGMA and ACPIM model 3 calculations for the AIDA experiments listed in Table 3. Best-fit values relate to the median 4 Fig. 6. Accommodation coefﬁcients αiceobtained from SIGMA and ACPIM model calculations for the AIDA experiments listed in Table 3. Best-ﬁt values relate to the median of the distributions of αice values that were retrieved by the uncertainty analysis described in Sect. 5.1, while error bars indicate the respective lower and upper quartiles. Likewise, median as well as lower and upper quartiles were determined from the temperature distribution of each experiment and applied accordingly. The temperature-averaged αice values obtained by SIGMA αSIGMA ice = 0.9+0.1 −0.5 and ACPIM αACPIM ice = 0.5+0.5 −0.3 are given by the solid black lines with the shaded region indicating their uncertainty. ice y y y Section 5.1 while error bars indicate the respective lower and upper quartiles. Likewise, 6 median as well as lower and upper quartiles were determined from the temperature 7 distribution of each experiment and applied accordingly. The temperature-averaged ice  8 values obtained by SIGMA SIGMA 0.1 ice 0.5 0.9     and ACPIM ACPIM 0.5 ice 0.3 0.5    are given by the 9 solid black lines with the shaded region indicating their uncertainty. 10 11 1 2 Figure 7. www.atmos-chem-phys.net/13/4451/2013/ Comparison of the combined SIGMA and ACPIM result 0.3 ice 0.5 0.7     (black solid 3 line, uncertainty illustrated by the shaded region) resulting from the AIDA cirrus cloud 4 Fig. 7. Comparison of the combined SIGMA and ACPIM result αice = 0.7+0.3 −0.5 (black solid line, uncertainty illustrated by the shaded region) resulting from the AIDA cirrus cloud experiments (purple crosses, cf. Fig. 6) between 190 K and 235 K with literature data from laboratory measurements (red symbols, cf. Table 1) and cirrus cloud model studies (blue bars, cf. Table 2). Error bars are included when given in the original publication. For the cirrus cloud model studies, the bars indicate the range of preferred αice values at one temperature or the preferred αice value within the considered temperature range, respectively. ice Section 5.1 while error bars indicate the respe 6 median as well as lower and upper quartile 7 distribution of each experiment and applied a 8 values obtained by SIGMA SIGMA 0.1 ice 0.5 0.9     and 9 solid black lines with the shaded region indicating 10 11 1 ctive lower and upper quartiles. Likewise, s were determined from the temperature ccordingly. The temperature-averaged ice  d ACPIM ACPIM 0.5 ice 0.3 0.5    are given by the their uncertainty. 2 Figure 7. Comparison of the combined SIGMA and ACPIM result 0.3 ice 0.5 0.7     (black solid 3 line, uncertainty illustrated by the shaded region) resulting from the AIDA cirrus cloud 4 Fig. 7. Comparison of the combined SIGMA and ACPIM result αice = 0.7+0.3 −0.5 (black solid line, uncertainty illustrated by the shaded region) resulting from the AIDA cirrus cloud experiments (purple crosses, cf. Fig. 6) between 190 K and 235 K with literature data from laboratory measurements (red symbols, cf. Table 1) and cirrus cloud model studies (blue bars, cf. Table 2). Error bars are included when given in the original publication. For the cirrus cloud model studies, the bars indicate the range of preferred αice values at one temperature or the preferred αice value within the considered temperature range, respectively. 40 1) and cirrus cloud model studies (blue bars, en in the original publication. For the cirrus of preferred ice  values at one temperature or mperature range, respectively. 0.003; Earle et al. J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice of αice (Magee et al., 2006; Earle et al., 2010; Isono and Iwai, 1969). The reason for these discrepancies can only be specu- lated upon at present. were at temperatures above 0 ◦C. In this case, frost built up on the inside of the experimental apparatus during the αice measurements may act as an unconsidered sink of water va- por and result in lower ice saturation ratios in the vicinity of the ice particles. The results of this work suggest that the ice particle growth in cirrus clouds is not signiﬁcantly impeded as it would be for a low value of the ice accommodation coefﬁcient αice < 0.1. An αice value close to unity also suggests that enhanced growth at few speciﬁc ice particle facets does not signiﬁcantly inﬂuence the observed ice particle growth that is governed by αice, i.e., up to a particle size of a few mi- crons. Implications of αice for cirrus clouds and their char- acteristics should therefore be minor. Furthermore, the result αice = 0.7+0.3 −0.5 is in good agreement with typical choices for αice in cirrus cloud modeling which lie in the range 0.2–1. Impact on prior calculations of cirrus cloud properties, e.g., in climate or regional models, is thus expected to be neg- ligible and future cirrus model studies can rely on a well- constrained ice accommodation coefﬁcient. If the given accuracy is assumed to be correct, it trans- lates into a minimum uncertainty of 25 % in supersaturation with respect to ice (Sice −1) since maximally obtained Sice in Magee et al. (2006) is around 1.2. Moreover, the initial size from which the ice particles in Magee et al. (2006) start to grow is above 6 µm at nearly ambient pressure conditions. Compared to the size of 0.7 µm from which ice growth is observable in our work, this should result in a rather low sensitivity with respect to αice. According to individual es- timates, this low sensitivity combined with the uncertainty in Sice would allow for αice values greater than 10−2. If the given uncertainty in Sice has been estimated too low, e.g., if water vapor losses to the wall of the experimental apparatus have been evaluated incorrectly, even much higher values of αice > 0.1 could possibly not be excluded by the experiments of Magee et al. (2006). Effect of latent heat of deposition When the release of latent heat from deposition of water molecules on the surface of an ice particle plays a signiﬁcant role, the ice particle surface temperature Ts is higher than the temperature Tg of the surrounding air. This results in an in- hibition of ice particle growth. The determining parameter in this context is the heat conductivity of air ka. As for the diffusivity Dw, ka has to be modiﬁed for gas kinetic effects which reduce the magnitude of ka for small ice particle sizes rp, cf. Eq. (3). The modiﬁed heat conductivity, k∗ a, is given by (Pruppacher and Klett, 1997; Eqs. 13–20) 7 Conclusions Phys., 13, 4451–4466, 2013 Atmos. Chem. Phys., 13, 4451–4466, 2013 J. Skrotzki et al.: Accommodation coefﬁcient of water molecules on ice Therefore, this we feel highlights the importance of in situ measurements of water vapor when at- tempting to quantify the ice accommodation coefﬁcient. www.atmos-chem-phys.net/13/4451/2013/ (2010) and Isono and Iwai (1969) retrieved αice values in the 10−2 range. A possible explanation for the very low values of αice obtained by these experiments could be the systematic underestimation of the ice saturation ra- tio Sice of water vapor. It has been pointed out in Sect. 5.1 that the uncertainty in the measurement of Sice usually is the dominant source for uncertainty in the retrieval of αice. An accuracy for Sice of 5 % is given in Magee et al. (2006) al- beit Sice is not directly measured in the region of ice particle growth but inferred from the amount of water vapor emitted from a water source into the experimental apparatus. Admit- tedly, it is argued that the humidity is calibrated using known growth factors for ammonium sulphate particles. However, it is not clear in Magee’s study whether these calibrations laboratory measurements (red symbols, cf. Ta 6 cf. Table 2). Error bars are included when g 7 cloud model studies, the bars indicate the ran 8 the preferred ice  value within the considered 9 10 relying on atmospheric in situ cirrus cloud data is the mea- surement of ice particle number concentrations which could yield artiﬁcially enhanced number concentrations due to shattering of cloud ice particles (Field et al., 2003; McFar- quhar et al., 2007). This enhanced ice number concentrations would result in an underestimated αice. However, this expla- nation does possibly not apply to the study by Gierens et al. (2003) in which 0.01 < αice < 0.1 was preferred. Gayet et al. (2006) argued that shattering of cloud ice particles can probably be excluded for the ﬁeld measurement data on which the analysis by Gierens et al. (2003) is based. 0 3 Discrepancies of the result αice = 0.7+0.3 −0.5 compared to lab- oratory measurements of αice are in part signiﬁcant. Magee et al. (2006) obtained the result 0.004±0.002 < αice < 0.009± www.atmos-chem-phys.net/13/4451/2013/ www.atmos-chem-phys.net/13/4451/2013/ Atmos. Chem. Phys., 13, 4451–4466, 2013 4463 7 Conclusions Dedicated experiments at the aerosol and cloud chamber AIDA were carried out to determine the accommodation co- efﬁcient of water molecules on growing cirrus ice particles, αice, in the temperature range between 190 K and 235 K. Pre- vious literature values of αice cover a range of almost three orders of magnitude in this temperature regime. Therefore, it is difﬁcult to assess the impact of αice on growth rates of ice particles in cirrus clouds, and consequently on ice parti- cle properties, number concentrations, and cirrus cloud radia- tive forcing. The experiments were conducted for a range of atmospheric conditions under which cirrus clouds typically form – including cooling rates and water vapor supersatu- rations. This resulted in realistic cirrus ice particle growth conditions yielding representative particle sizes and shapes. k∗ a ka = 1 1 + 4ka αTrpρacp,a ¯cw , (A1) (A1) where αT is the thermal accommodation coefﬁcient, ρa the density of air, cp,a the speciﬁc heat of air, and ¯cw the mean thermal speed of air molecules corresponding to Eq. (4). The thermal jump distance 1T was set to zero according to Fukuta and Walter (1970). As choice for αT, experimental results suggest a value of unity (Mozurkewich, 1986). The data sets of the performed AIDA experiments were in- dependently evaluated by two different models – SIGMA and ACPIM. With these models it was possible to retrieve a best- ﬁt value for the ice accommodation coefﬁcient αice along with uncertainty bounds for each individual experiment. No signiﬁcant temperature dependence of αice was observed. The temperature-averaged value resulting from the SIGMA model is αSIGMA ice = 0.9+0.1 −0.5. This result is in good agreement with the independent analysis by the ACPIM model yield- ing αACPIM ice = 0.5+0.5 −0.3. The combined result of both mod- els, αice = 0.7+0.3 −0.5, compares well with most of the previous model studies of cirrus ice particle growth in the atmosphere or in cloud chambers. There are, however, signiﬁcant dis- crepancies with respect to three relevant laboratory retrievals Taking the effect of latent heat release into account yields for the latent heat term LH in Eq. (6) LH = Ls k∗aTg LsMw RTg −1 . 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tel-04217850-Th%C3%A8se%20Baticle%20Christophe.txt_45	French-Science-Pile	Open Science	Various open science	2,023	Les pratiques de chasse comme affirmations politiques du principe d'autochtonie : Dimensions territoriales des luttes cynégétiques : Picardie, Normandie, Pyrénées, Savoie. Sciences de l'Homme et Société. Université de Picardie Jules Verne, 2007. Français. ⟨NNT : ⟩. ⟨tel-04217850⟩	None	French	Spoken	7,183	11,314	Si bien que tant la Baie de Somme est le terrain des sauvaginiers du département, tant bcelle de la Baie d’Authie se partage avec les résidents du Pas-de-Calais, voire serait quel que peu leur déversoir quand la partie nord de la même baie n’accueille que quelques huttes et celle de la Canche, plus au nord, reste limitée. Une vue rétrospective de l’extériorité en Baie d’Authie De manière à vérifier cette hypothèse d’une désaffection en Baie d’Authie, conséquence du caractère exogène de bon nombre de sociétaires, nous avons pu reconstituer la répartition par département de résidence sur la longue période. Il apparaît ainsi que l’on vient de loin pour 962 Nous ne disposons pas en effet de la même exhaustivité, bien que l’amplitude temporelle soit identique, en ce qui concerne cette ACDPM que pour ce qui concernait sa collègue de la Baie de Somme. 963 La phase de décollage y connaît la même pente ascendante, mais accentuée, de la même manière que le reflux de la fin des années 80 ou le plateau de la seconde moitié de la décennie 90. 611 chasser en Baie d’Authie, même si c’est en nombres réduits. La limitation des accès terrestres lui confère un charme sauvage, prisé des chasseurs recherchant le calme et l’isolement (même relatif). CAMPAGNE Départements* 25 32 33 37 34 44 52 Départements les plus représentés Nord/Oise/Seine-St-Denis Nord/Oise/Paris Nord/Seine-St-Denis/Val-de-Marne Nord/Seine-St-Denis/Val-de-Marne Nord/Seine-St-Denis/Val-d’Oise Nord/Seine-St-Denis/Oise Nord/Seine-St-Denis/Oise % « extérieurs »** 42,6% 44,6% 43,6% 41,3% 38,4% 37,5% 37,5% 1975-1976 1976-1977 1977-1978 1978 -1980 1980-1981 1981-1982 2000-2001 31 Nord/Oise/Val-d’Oise 20 ,6% * Hors Somme et d épartement limitrophe du Pas-de-Calais . ** % représentant l ’ensemble des « extérieurs » sur l’ensemble des effectifs inclu ant la Somme et le Pas-de-Calais. D’emblée, les résidents des départements extérieurs à la Somme et au Pas-de-Calais ont représenté une part substantielle des effectifs adhérents de l’ACDPM, avec une croissance dès la seconde année qui correspond à la forte augmentation constatée globalement. A partir de cette période la chute de la part exogène est continue jusqu’à ne plus atteindre qu’un cinquième du total. Pour exemple, alors que l’on comptait encore 711 « extérieurs » pour la saison 1976-1977, ils n’étaient plus que 183 lors de la campagne 2000-2001. De ce fait, on peut valider que cette composition géographique spécifique en Baie d’Authie explique une part de sa considérable régression. Baisse qui touche un peu plus Paris et l’Ile-de-France, cette région passant de 23,7% de l’ensemble en 75-76 à 19,1% en 81-82. Les chasseurs franciliens ne représentaient en 00-01 que 6,6% des sociétaires. Evolutions Comparées Ile-de-France "Extérieurs" 80 & 62 Ensemble 2500 2000 1500 1000 500 0 75-76 76-77 77-78 78-79 79-80 80-81 81-82 Concernant maintenant les codes postaux, on en dénombre 264 distincts pour cette association, dont 88 relevant du Pas-de-Calais et seulement 36 de la Somme. La dispersion s’avère forte pour ces voisins nordistes, le premier code postal (Berck) ne regroupant que 41 adhérents, très loin derrière Rue, en tête pour la Somme et tous départements confondus avec 186 sociétaires 612 Ainsi, même s’ils sont minoritaires dans l’ensemble, lorsqu’ils résident dans la Somme ce sont généralement des locaux au sens du canton de Rue et particulièrement les communes directement en façade côtière964. Afin de comprendre les principaux motifs de l’érosion démographique dont sont l’objet les associations du DPM, nous disposons également de deux enquêtes qui permettent de cerner quelques éléments de réponses. La première de ces études concerne justement l’ACDPM de la Baie d’Authie965. Elle fut menée par le biais d’un questionnaire envoyé par la voie postale et concernait les adhérents 2000-2001 n’ayant pas renouvelé leur carte en 01-02. Sur 310 envois, les 147 questionnaires retournés, dont 130 exploitables (42%), font état d’un contexte juridique contribuant pour une bonne part à l’explicitation des motifs. Dans l’ensemble il apparaît que des conditions plus limitées de la pratique, liées à un public parcourant une longue distance afin de se rendre en Baie d’Authie, soient les principales causes de la déperdition dans cette association. C’est donc pour une grande part le type géographiquement éclaté du recrutement qui apparaît comme ici déterminant en toute vraisemblance. On assiste en conséquence à une relocalisation dans le renouvellement des effectifs. 3.) L’ACD PM du Littoral Picard-Sud : la plus forte dé croissance chez les « chasseurs aux pieds nus » Cette troisième et dernière ACDPM picarde a pour particularité de ne pas couvrir un estuaire, mais de courir le long de 22 km de côtes, offrant 900 ha de domaine public. Ils ont parfois pu se retourner sur l’ACPH du hâble d’Ault966, dont le territoire est contigu. Néanmoins, si l’on a pu dire, dans les années soixante, à propos de l’immense citéusine Renault « Ne désespérez pas Boulogne-Billancourt » pour signifier le chaudron social que représentait alors la « citadelle ouvrière », la même formule appliquée au Vimeu Rouge fait facilement éch chez les dirigeants cynégétiques départementaux. Et effectivement « ches durs d’ch’ Vimeu »967 ont pour réputation de défendre pied à pied leur passe-temps favori. 964 Les dix communes les plus densément pourvoyeuses le sont moins qu’en Baie de Somme, mais ce sont surtout Fort-Mahon et Quend-Plage qui se détachent. Ces deux villes se caractérisant par une importante population résidentielle, souvent en provenance d’autres départements, il est crédible de penser que l’effectif exogène s’avère plus important que ce qu’il en apparaît dans le listing, ces adhérents ayant pu indiquer leur résidence secondaire comme lieu d’habitation à saisir. 965 A.C.D.P.M. Baie d’Authie-Sud : Enquête sur les déperditions entre les campagnes 1999-2000 et 2000-2001, février 2000. 966 Association des Chasseurs, Propriétaires, Huttiers, présidée par le premier représentant de l’ACDPM Littoral Sud, les identités sociogéographiques des deux structures s’avèrent très proches. 967 Expression reprise d’un chasseur amiénois dans le car qui nous menait à Paris pour la grande manifestation de 1998, alors qu’arrivaient à notre hauteur les petits bus affrétés par les associations de chasse du Vimeu. Au regard des 4,7% de perte annuelle moyenne depuis la saison 1978-1979, ce que d’aucun appelle « l’irrédentisme du Vimeu » peut également apparaître comme une réaction à la désaffection des effectifs, signe d’une perte de puissance notable. Surtout, nous sommes ici face à une chasse de spécialistes très spécifique, exigeant un savoir-faire et une connaissance tant du milieu que des espèces et de leur calendrier, qui associe avec une rare précision les conditions météorologiques du moment à la topographie des lieux. Ces « chasseurs aux pieds nus », pour reprendre l’expression d’un connaisseur de la presse cynégétique, ont développé une fierté de leur ingéniosité inversement proportionnelle à la raréfaction de leurs effectifs. Il s’agit en effet d’une quête plutôt orientée vers les limicoles, soit ces oiseaux vivant dans et de la vase. Afin de les amener à se poser dans l’aire retenue par le chasseur celui-ci se doit de ne faire aucune erreur quant aux « formes » (ou « blettes », nom picard ce ces leurres) retenues. Dos au rivage, ce nemrod particulier ne doit pas attendre d’attirer un oiseau autre que les congénères en plastique ou en bois qu’il a disposé sur la plage. Le chiffre de 1 100 adhérents est souvent avancé localement pour quantifier le point culminant atteint par l’association. Pour ce qui nous concerne, nous pouvons avaliser celui de 943 en 1985-1986. Quoiqu’il en soit, le Littoral Sud a vu le départ d’un nombre considérable de licenciés et connaît actuellement les effectifs les plus bas de son histoire. Ils ne représentent plus qu’un peu plus du cinquième de ce qu’ils étaient en 1985-1986 : 206. Ici encore et comme pour sa voisine de la Baie de Somme, un moment-clef marque un revirement de tendance avec la seconde partie de la décennie quatre-vingt. Alors que sur la durée précédente les effectifs vont plutôt croissants, avec en moyenne et par an plus 4,3%, l’ère la plus proche de nous se trouve très affectée par une érosion sans commune mesure avec la décroissance des validations du permis de chasser en France sur la période (-1,78% en moyenne et par an de 1985-1986 à 2000-2001. Il n’y a guère eu que la saison 1996-1997 pour contredire cette baisse quasi continue, les années suivantes ayant absorbé la recrudescence temporaire. 00 -0 1 98 -9 9 96 -9 7 94 -9 5 92 -9 3 90 -9 1 88 -8 9 86 -8 7 84 -8 5 82 -8 3 80 -8 1 78 -7 9 1000 900 800 700 600 500 400 300 200 100 0 S AIS ONS Une telle décrue s’explique par un ensemble de phénomènes, dont les périodes de chasse occupent la première place. Avec la rétraction de la période estivale, nombre de résidents extérieurs à la région ont, comme en Baie d’Authie, renoncé à prendre leur carte. De plus, les caractéristiques topographiques du site rendant impossible l’installation de huttes, seules les chasses au hutteau et à la botte y sont pratiquées. De ce fait, l’attractivité du littoral Sud peut s’avérer moindre pour les amateurs de canards. Pourtant la richesse en limicoles a longtemps contrebalancé ce handicap. Les mesures de protection pour ces derniers ont alors 614 fait pencher la balance dans l’autre sens. De plus, l’accentuation de la pression touristique pour ces « hauts lieux » du littoral a globalement été inversement proportionnelle à la pression cynégétique. Enfin, les récents travaux sur la digue ont causé des perturbations pour la pratique, amenant certains chasseurs à se replier sur d’autres territoires. Pour finir, il est permis de s’interroger sur les difficultés propres à un mode de chasse aussi spécifique quant à sa reproduction sociale. Effectifs sexués : quid des femmes? Seules cinq femmes sembleraient adhérer à cette association, ce qui en ferait néanmoins l’association la plus féminisée des ACDPM. du littoral picard avec 2,4% des sociétaires. Même si cela reste peu, ce constat nous amène à deux remarques, d’importance nous semble-t-il. Primo, domaine « public » ne signifie en rien mixité, et ce en conformité avec tous les acquis de l’anthropologie quant à la division sexuée de l’espace. Secundo, la sociologie classique pousserait à penser que la cynégéphilie d’une région jouerait logiquement dans le sens d’un plus large accès des femmes aux territoires de chasse, ce qui se trouverait alors ici vérifié. Mais toujours en phase avec ce classicisme, les spécialistes de la chasse s’accordent souvent pour reconnaître un clivage social déterminant en matière de mixité : les femmes des classes populaires étant plutôt invitées à s’abstenir d’entrer dans le cercle des activités viriles, tandis qu’à l’autre extrême de l’échelle sociale, en particulier dans l’aristocratie, cette exclusion tacite aurait un caractère moins clivant. Il semblerait ici qu’il faille revisiter cette question pour chercher à vérifier l’hypothèse selon laquelle ce serait les deux pôles de la stratification sociale qui faciliteraient le plus l’entrée des femmes dans la chasse. Il faudrait dans ce cas s’interroger sur ce qu’un sociologue comme Pierre Bourdieu appelait « l’ethos de classe », à savoir une éthique pratique, non dogmatique et inconsciente, qui s’incorpore dans les êtres, par exemple ce qu’il convient de faire ou de ne pas faire quand on est une femme « respectable » pour son milieu social. Il y aurait ici sûrement fort à dire sur les dispositions à la pratique cynégétique qu’offre l’ethos populaire dans un contexte d’effacement des distinctions sexuées. Encore que pour ce qui concerne les baies, leurs marées rendent la fréquentation cynégétique particulièrement éprouvante, renforçant probablement encore pour longtemps la quasi exclusive masculine. Par ailleurs, la stratégie actuellement favorisée par les instances cynégétiques va dans le sens d’une ouverture sur les femmes, non seulement parce qu’elle permet de lutter contre l’image machiste qui colle à la chasse, mais aussi dans le but de relancer des adhésions qui se tarissent durablement. Le même phénomène fut observé pour de nombreuses activités dont le point commun est le primat du physique, qu’il s’agisse de force pure ou d’endurance 968. Il reste que l’Association Nationale des Femmes Chasseresses (« Chasser au Féminin ») fut fondée et se trouve présidée par une femme issue de notre département et du milieu des sauvaginiers du littoral picard. « Gens d’ici » : une autochtonie affirmée A rebours de l’ACDPM Baie d’Authie, le littoral Sud a pour réputation de constituer un milieu plus endogène, où la déclinaison localiste du cogito cartésien par Guy Di Méo prend plus d’importance qu’ailleurs : « Je suis, donc je suis de quelque part » et plus particulièrement « d’ici ». Le nombre de départements y est en effet peu élevé (9 seulement), mais surtout, si l’on prend en considération la situation géographique quasiment limitrophe du territoire associatif avec la Haute-Normandie, on obtient une presque exclusivité des résidents de la Somme et de Seine-Maritime (près de 92%), l’Ile-de-France ne comptant ici que pour 968 Pour n’en donner que des exemples régionaux, le Ballon au Poing, présent dans la région d’Albert, ou la Longue Paume, répartie dans le Santerre Somme et le bassin d’emploi défini par l’INSEE en « Santerre Oise », ont procédé de même. 615 moins de 3,5%. Même en matière de codes postaux, leur nombre élevé (24) ne dissimule pas la prédominance de l’arrondissement d’Abbeville. Par rapport aux effectifs maintenant, le Vimeu, en particulier dit « industriel », se retrouve majoritaire. Les ouï-dire quant à la dimension localiste des adhérents de cette ACDPM trouvent ainsi à se vérifier au regard de ces données. Un constat identique peut se tirer pour les dix premières communes, toutes dans un périmètre de quelques kilomètres au plus, nouveau signe d’un primat des locaux parmi les adhérents de cette association. Cette situation de déperdition explique qu’on en ait tenté l’analyse via une étude spécifique969, laquelle révèle un effet pécuniaire plus sensible qu’ailleurs, rebondissant en quelque sorte de la composition plus populaire du chaudron vimeusien. Ici plus encore qu’ailleurs, le coût de l’activité, associé au contexte légal et aux difficultés liées à la superposition des publics sur ce territoire sont des éléments déterminants, soit pour renoncer à la chasse associative du gibier d’eau, soit pour adhérer à une autre structure. 4.) L’Association des Chasseurs Propriétaires Huttiers du Hâble d’Ault : un chaudron social en forme de soupape d’échappement Bien qu’il ne s’agisse pas d’une ACDPM et que son périmètre n’entre pas totalement dans le site Natura PIC.01, nous avons choisi de présenter quelques données de l’Association des Propriétaires Huttiers du Hâble d’Ault, dans la mesure où comme précisé plus haut cette structure sert de déversoir par rapport aux problèmes de déficit territorial sur le DPM. Pour autant, ces chiffres à but uniquement informationnel ne seront pas inclus dans la synthèse qui suit et qui concerne l’ensemble des trois associations publiques. Pour le dernier listing disponible, on y comptait 283 adhérents inscrits970, soit sensiblement plus que l’effectif de l’ACDPM Littoral Sud. On y dénombre également peu de femmes malgré le caractère non exclusivement cynégétique de l’adhésion : 3,2%. Ainsi, même lorsque l’adhésion n’est pas liée à la pratique de la chasse, ce sont des hommes qui s’impliquent le plus dans les associations cynégético-foncières. Quant à l’inscription territoriale, nous avons observé que le nombre de résidents de la Somme allait croissant de la Baie d’Authie (30,21%) au Littoral Sud (83,50%) en passant par la Baie de Somme (82,36%). On atteint avec l’ACPH un nouveau rapport proche des plus forts taux. Ils sont en effet plus de huit sur dix alors que le nombre de départements représenté y est le plus faible avec 969 ACDPM Littoral Picard-Sud : Enquête sur les déperditions entre les campagnes 1995-1996 et 1996-1997, novembre 1996. La collecte des données a là aussi été réalisée sur la base d’un questionnaire envoyé par voie postale et donc auto-administré par les enquêtés. La population visée concernait les sociétaires de la saison 19951996 n’ayant pas renouvelé leur adhésion pour 1996-1997, soit 95 individus. A cette époque, l’association est en train de connaître sa plus spectaculaire remontée (+ 33,4%) après une décennie de décroissance globale dont une année seulement en stagnation. 970 Deux remarques préliminaires doivent cadrer la valeur des effectifs concernant l’ACPH : primo l’adhésion n’y est pas obligatoire, ce qui induit à l’évidence un certain déficit de données et secundo on y compte quelques adhérents « sympathisants », voire inscrits à titre honorifique pour services rendus à l’association. De plus, s’agissant également d’un regroupement de propriétaires, tous ne sont pas personnellement impliqués dans la chasse. 616 seulement sept. Quand ils ne sont pas habitants de la Somme c’est dans un département limitrophe qu’ils résident souvent (13%), alors que moins de 2% se situent au-delà. On enregistre néanmoins quelques 60 codes postaux, ce qui laisse imaginer une plus grande dispersion géographique qu’il n’y paraîtrait au premier abord. Parmi ces 60 codes 36 concernent la Somme, soit beaucoup plus que les 24 de l’A.C.D.P.M. Littoral Sud, ce qui révèle une dispersion géographique beaucoup plus importante au sein du département. 17 codes sont communs aux deux associations, mais ceux qui sont propres à l’A.C.P.H. se situent parfois à l’autre extrémité du département, comme Roye, Combles, Ham ou Roisel. On a par là même un des signes de cette incitation plus sensible à passer de l’ACDPM à l’ACPH pour les moins disposés à se rendre sur le littoral sans l’assurance que des conditions météorologiques défavorables n’obligeront pas à rebrousser chemin. C’était moins vrai lorsque la saison estivale commençait au 14 juillet et c’est en quelque sorte compensé par ce pied-à-terre qu’offre la hutte du Hâble d’Ault. Rappelons ici que ce débat sur les dates de la pratique débouche sur des conséquences parfois inattendues en matière d’occupation territoriale. De ce fait, en dehors d’Abbeville, qui arrive à la huitième position et du Tréport en dernière, les dix premières communes de résidence bordent quasiment le hâble. On a probablement ici un effet de la propriété. Le reste de la liste confirme d’ailleurs ce point. Enfin, le listing nous informe sur certaines des motivations de l’adhésion, dont la détention du permis de chasser qui concerne 184 membres (65,02%). Les numéros de ces permis permettent encore d’en connaître les départements où ils furent acquis. Des numéros qui font état de dix départements, l’ordre des départements restant identique à celui tous adhérents confondus, avec une Somme très largement en tête devant la Seine-Maritime. A l’évidence, la chasse constitue l’objet majeur de l’adhésion à l’ACPH. 5.) Une fiche d’identité971 des chasseurs du DPM picard Avec 3 613 droits de chasser pour la période de l’enquête972, l’évolution sur la longue durée fait état de moins de 2% de pertes en taux de variation annuel moyen depuis la campagne 1978-1979, soit une régression apparemment moindre à ce qu’il en fut pour les chasseurs de France. Néanmoins, une décroissance presque continue se profile dès la saison 86-87 et caractérise le long terme, sauf pour la campagne 96-97, période pendant laquelle on constate une légère remontée. La perte globale s’élève alors à plus de 37% pour l’ensemble de l’étendue, avec deux phases, la décrue allant en s’accentuant de la première à la seconde. Pour cette dernière, les ACDPM ont atteint un taux de décrue plus de quatre fois au chasseur français (-6,1% de 96-97 à 00-01 contre -1,5%). Surtout, elles ont perdu plus de la moitié de leur optimum atteint en 85-86 avec 7 254 sociétaires. Notons également que la saison 00-01 fut une campagne particulièrement négative, après que le parlement ait refondu le droit français en matière de chasse. 971 Précaution d’emploi : les chiffres donnés dans ce point ne correspondent pas à des individus, mais bien à des droits de chasser. Certains chasseurs ont en effet une double, voire triple adhésion aux ACDPM étudiées. Si l’on retient les résultats de l’enquête « Opinion » (détaillée plus loin) et si l’on considère l’échantillon représentatif de l’ensemble de la population, les double-comptes représentent 14,6% des effectifs cumulés. 972 Hors double-comptes maintenant, en appliquant le taux de 14,63% rencontré dans l’enquête Opinion des chasseurs du DPM, on obtient 3 024 chasseurs, lesquels remplacent ici les précédents « droits de chasse » et constituent un effectif « redressé ». 617 Somme Sud Ces trois associations ont vécu des évolutions globalement similaires, mais avec des décalages temporels d’une à deux années pour les retournements d’importance. Il subsiste que si cette décroissance globale est tributaire d’un contexte général de crise, elle correspond aussi à l’émergence du « parti des chasseurs ». En 8 99 -0 6 97 -9 4 95 -9 2 93 -9 0 91 -9 8 89 -9 6 87 -8 4 85 -8 2 83 -8 0 81 -8 8 79 -8 6 75 -7 77 -7 4500 4000 3500 3000 2500 2000 1500 1000 500 0 Quant à la composition des trois associations prises toujours dans leur ensemble, les droits de chasser se concentrent surtout sur dix départements pour 96% d’entre eux. La Somme approche les trois quarts et plus de 85% avec le Pas-de-Calais. En-déca, les autres départements pourvoient pour des valeurs très inférieures. Même l’Ile-de-France ne recueille ici qu’à peine plus de 5%, alors qu’elle fut depuis longtemps considérée comme une grande pourvoyeuse de nemrods pour le littoral picard. Il n’est donc pas possible de prétendre, comme on l’entend parfois, que la population cynégétique locale tendrait à devenir de plus en plus distante d’avec les terrains de chasse. C’est plutôt au contraire à un recentrage sur la Somme auquel nous assistons. Par contre, une règle toute géographique semble se confirmer avec une domination des habitants du départements, suivis par les limitrophes (23,7%), puis par les franciliens. Pas-de-Calais 15% Oise 3% Nord Seine-Maritime 3% 2% Val-d'Oise 2% Seine-et-Marne 1% Yvelines 1% Seine-Saint-Denis 1% Hauts-de-Seine 1% Somme 71% La liste des dix premiers codes postaux reprend ici quasiment celle de l’ACDPM de la Baie de Somme, dans la mesure où il s’agit de la plus peuplée. Seul l’ordre est quelque peu modifié par l’association de la Baie d’Authie qui fait remonter Rue de la quatrième à la seconde position, et le Littoral Sud qui fait gagner trois places à FrivilleEscarbotin (de la dixième à la septième). De plus et surtout, la prédominance des communes du PIC.01, si l’on ne retient que ces dix premiers codes, est massive puisque ce sont plus de la moitié des chasseurs qui en relèveraient, plaçant résolument les chasseurs du DPM parmi les riverains côtiers. 618 6.) Les « croyants et pratiquants » de Rue : « le meilleur canton de France » En préambule, rappelons ici qu’il convient de préciser que l’adhésion à la société communale des Chasseurs de Rue (comme à toute association communale ou ACDPM) permet quant à elle l’exercice de la chasse sur son territoire, contrairement à l’ACGER, dotée d’un statut davantage syndical. Pour cette dernière, 137 particuliers lui accordaient leur adhésion en juin 2003, alors que l’association elle-même en annonçait une centaine dans le guide du sauvaginier de la Somme édité pour la campagne 04-05. En d’autres termes, elle aurait subi une certaine désaffection, l’enquête de terrain ayant vérifié qu’elle avait en effet compté dans le passé beaucoup plus de membres, certaines difficultés liées à la succession d’un au charisme inégalé ayant contribué à cette décrue. Créée dans les années cinquante, elle a réuni jusqu’à 268 adhérents dans les années quatre-vingts, alors qu’on y comptait encore un nombre plus important de résidents extérieurs au canton même de Rue, ainsi que davantage de propriétaires de huttes, lesquels ont peu à peu déserté l’organisation. Pour expliquer ce désintérêt, on peut avancer en plus de la notoriété de l’ancien président, notable jouissant d’une réputation indiscutable, le service rendu en matière d’achat de colverts pour le repeuplement des marais. Par ce dispositif, les adhérents bénéficiaient de tarifs préférentiels. Par ailleurs et plutôt, la dimension extrêmement populaire du recrutement, que nous analyserons plus loin, a pu laisser le jeu de la distinction sociale s’exprimer. De plus, la disparition progressive des sociétés communales de chasse du secteur a indéniablement atomisé les chasseurs locaux, ce qui n’a pas favorisé leur regroupement au sein d’une instance de défense. En perdant les plus motivés des adhérents d’autres communes, l’effet « boule de neige » s’est estompé, provoquant un recentrage sur la ville de Rue. Enfin, la systématicité de la double adhésion (ACGECR/Société Communale) si elle reste prégnante, a perdu de son efficacité, alors que dans le même temps cette Société a perdu de ses effectifs. Aujourd’hui, le niveau cantonal poursuit ses tentatives en direction des propriétaires « parce qu’ils ont du poids », mais sans le succès escompté : « Quand tu vois qu’un type comme X il nous refuse une cotisation à 7€! » Ce syndicat, à ses origines section locale de l’ANCGE, a également subi les derniers revers électoraux de CPNT dont il était le meilleur promoteur localement , lui offrant son meilleur canton lors des européennes de 1999, mais encore à l’occasion du premier tour de la présidentielle de 2002. Au sein des ACDPM la gent féminine s’avérait déjà très peu représentée avec une proportion de l’ordre de 1,3%, mais ici et malgré le caractère non exclusivement cynégétique de l’association, puisqu’il s’agit plus de défendre que de seulement pratiquer, le taux de virilité devient écrasant avec une unique femme recensée. S’agissant d’une association qui comporte dans son intitulé même la vocation locale de son objet, on peut se demander si elle attire les seuls résidents du secteur cantonal, ou audelà les pratiquants du canton qui n’y habitent pas, lesquels témoigneraient ainsi d’un souci pour la défense de leurs terrains de chasse, en dehors des questions du vécu quotidien. De plus, les associations ont pleinement conscience des avantages stratégiques qui tiennent dans un recrutement large dépassant les limites strictes de leurs aires d’action, en mettant en avant 619 l’attractivité dont témoignent ces adhésions extra locales. Concernant donc le département d’habitat et à l’exception de deux ressortissants du Pas-de-Calais, seule la Somme est citée. Et encore, ce 1,5% de « 62 » concerne le sud du département voisin, avec la région d’Aubigny. A lui seul, le canton de Rue rassemble près de neuf cartes sur dix. Néanmoins, près de 13% des adhérents proviennent des alentours, validant une forme d’exogénéité du recrutement. Dans ces cas de figure, les deux sites Natura 2000, au sens des codes postaux attribués aux trente communes les composant, représentent la presque totalité des adhésions. De même, si l’on s’attache aux communes de résidence, on constate que l’ensemble de la population est composée à82,5% d’habitants des communes du PIC.02. En interrogeant nous ciblerons ainsi des individus dont les localités de résidence sont parfaitement incluses dans le terrain retenu. A propos des propriétaires huttiers que nous évoquions ci-dessus, ils restent présents avec seulement moins de 6% des membres (huit adhésions), ce qui s’avère effectivement très peu. Au-delà de cet engagement sur le canton de Rue, 17% des adhérents relèvent également d’une ACDPM, dont la majeure partie en Baie de Somme. En vis-à-vis des désavantages du nombre, l’ACGECR permet à ses dirigeants d’en connaître très bien les membres, y compris sur le plan professionnel. Les entretiens ont de ce fait permis d’établir une classification précise sur ce plan. On constate à ce niveau une majorité relative d’ouvriers qui avoisine les 50%, alors que les agriculteurs s’y montrent peu nombreux, ainsi que les classes dites « supérieures » totalement absentes. CLASSIFICATION APRÈS RECODAGE DES 28 RETRAITÉS Profession Indéterminé Agriculteur-Exploitant Contremaître, Agent de Maîtrise etc. Ouvrier Dont Ouvrier Agricole Employé Artisan, Commerçant Cadre Moyen Cadre Supérieur Chef d’Entreprise Profession Intermédiaire Profession Intellectuelle Sans Activité Dont étudiant TOTAL DES OBSERVATIONS Nb. Cit. 11 6 0 81 10 5 8 4 2 0 6 4 10 2 137 Fréq. 8,0% 4,4% 0,0% 59,1% 7,3% 3,6% 5,8% 2,9% 1,5% 0,0% 4,4% 2,9% 7,3% 1,5% 100% Seconds en effectifs les retraités et pré-retraités donnent maintenant une majorité absolue à la PCS ouvriers, qu’on trouve dans près de six cas sur dix. En revanche, les artisans et commerçants, pourtant nombreux dans les rangs cynégétiques, entretiennent ici un effectif assez confidentiel. Le caractère populaire de l’ACGECR est en conséquence validé, les classes laborieuses y étant majoritaires de façon écrasante. 620 7.) La Société Communale de la ville de Rue : une jumelle qui recrute large Comme pour l’ACGECR, la Société Communale n’accueille qu’une seule femme, et encore qui ne réside pas dans le secteur. C’est la première situation de ce type rencontrée sur les deux enquêtes pour une association qui regroupe autant de sociétaires. Bizarrement également, alors que ses statuts en font une association communale et qu’elle comporte moins d’adhérents (105) que l’ACGECR, la Société de Rue compte davantage de résidents du Pas-de-Calais La proportion des habitants du canton s’y est légèrement tassée, laissant percevoir que cette association recrute davantage à l’extérieur qu’une banale société communale, ici jusque dans le Santerre. En ce qui concerne maintenant les communes, si Rue arrive logiquement en tête avec 78%, c’est en revanche la seule commune du PIC.02 représentée sur les seize agglomérations répertoriées. Quant aux propriétaires huttiers, leur effectif déjà faible au sein de l’ACGECR s’effondre ici avec uniquement trois membres. Enfin, plus d’un sociétaire sur cinq émarge également dans une ACDPM et toujours plutôt en Baie de Somme. La proximité entre adhérents cantonaux et sociétaires de Rue n’est pas que géographique, mais aussi et surtout sociale. La seule différence notable entre les deux associations tient dans la proportion moins importante de retraités dans les rangs de la Société. Il est effectivement courant de trouver davantage d’actifs dans les chasses communales, dont les tarifs sont souvent modérés. Arrivés au terme de leur activité professionnelle, certains chasseurs s’investissent pour la première fois dans des domaines privés faisant appel à l’actionnariat, généralement plus onéreux. C’est probablement le signe d’une disponibilité plus importante, mais aussi de la diminution des dépenses d’investissement qui marquent la fin de vie, laissant plus de ressources pour les loisirs. Reste qu’on y trouve presque autant d’ouvriers et de représentants des classes populaires après recodage des retraités. Ce sont les agriculteurs-exploitants, la plupart du temps détenteurs de droits de chasse qu’ils peuvent se réserver, qui régressent sensiblement par comparaison avec ce qu’ils représentaient au sein de l’ACGECR. 8.) L’identité des chasseurs de base sur l’arrière litt oral Enfin, comme pour les ACDPM précédemment, le cumul des deux associations, ACGECR et Société Communale de Rue, à l’exclusion des doubles comptes, donne l’ensemble de la population pour la seconde vague de questionnaires. La féminité s’y retrouve logiquement très faible avec seulement deux adhérentes, soit 1,8% des 170 membres de la population interrogée. Seuls six codes postaux sur onze ne relèvent pas du département, mais du Pas-deCalais limitrophe pour sept adhérents (4,1%), tout comme se démarque le canton de Rue avec la plus forte concentration de cartes, soit près de 85%. Sur vingt-sept communes de provenance, vingt relèvent de la Somme et sept du Pas-de-Calais. Parmi les personnes interrogées, seules huit sont propriétaires huttiers et avec près d’un quart des effectifs membre d’une ACDPM, le gibier d’eau semble occuper une part déterminante dans leurs orientations cynégétiques, d’autant que nombre de ces adhérents peuvent pratiquer cette forme de chasse en marais d’intérieur, à Rue même. Lorsque c’est sur le domaine maritime on constate que c’est le plus souvent en Baie de Somme 28/40). La répartition par profession ou groupe de métiers nous intéresse au premier chef parce qu’elle permet de fournir l’identité sociale dominante de la population, mais d’un point 621 de vue plus méthodologique elle fournira une indication qui servira à vérifier la représentativité de l’échantillon répondant au questionnaire. CLASSIFICATION APRÈS RECODAGE DES 30 RETRAITÉS Profession Indéterminé Agriculteur-Exploitant Contremaître, Agent de Maîtrise etc. Ouvrier Dont Ouvrier Agricole Employé Artisan, Commerçant Cadre Moyen Cadre Supérieur Chef d’Entreprise Profession Intermédiaire Profession Intellectuelle Sans Activité Dont étudiant TOTAL DES OBSERVATIONS Nb. Cit. 21 8 0 91 11 7 8 6 3 0 7 4 15 4 170 Fréq. 12,4% 4,7% 0,0% 53,5% 6,5% 4,1% 4,7% 3,5% 1,8% 0,0% 4,1% 2,4% 8,8% 2,3% 100% Alors que la région de Rue se caractérise plutôt par la prédominance des services et du secteur primaire, on observe chez les chasseurs retenus pour l’investigation des tendances très nettement orientées vers le secondaire. En effet, même si 47% des hommes vivant dans la Somme relèvent de ce secteur, ils sont moins de 30% sur le canton qui nous intéresse en premier lieu, la dimension populaire de la population d’enquête restant très affirmée. B.) CONSTITUTION DE L’ÉCHANTILLON : recherche de la représentativité optimale Les listings des cinq associations visées par les deux vagues de questionnaires lancés par site Natura disposent donc de fichiers nominaux, comportant patronymes, prénoms et adresses des adhérents, ce qui a permis un échantillonnage par ACDPM, selon le sexe, le département et le code postal. Par ailleurs, nous avons cherché à établir une proportionnalité par commune pour celles qui pourvoient le plus à l’effectif global des chasseurs recensés dans ces associations973. A partir de ces préalables méthodologiques, il a été décidé de viser un sociétaire sur cinq. Au total, ce furent ainsi 800 questionnaires d’expédiés. En revanche, la méthode fut simplifiée en ce qui concerne les deux associations du PIC.02, leurs effectifs permettant d’interroger l’ensemble des membres. Taux d'enquête sur les ACDPM : 22,14% de la population-mère, soit 800 questionnaires Taux d'enquête sur les associations de Rue : 99% de la population-mère, soit 169 questionnaires 973 Recherche également du questionnaire unique par foyer connaissant les redondances de ce côté et la concertation entre apparentés pour répondre, ce qui laisse espérer un taux d’enquête réel supérieur ; élimination des doublons (cas de figure d'un pratiquant relevant de plusieurs ACDPM). 622 1.) Taux de participation : une volonté forte d’expression qui interroge quant au rôle cathartique de l’enquête d’opinion Au - delà de cette recherche de représentativité, les bases de données disponibles ne nous ont pas permis de prendre en considération d’autres déterminants classiques fondamentaux de l’opinion telle que définie en sociologie, en particulier l’âge, la ruralité ou l’urbanité du lieu de résidence, mais surtout la profession. Seule une étude, réalisée sur ’unique ACDPM Baie de Somme permettait d’apprécier cette dernière variable, mais déjà ancienne et sans liaison avec le fichier des adresses. Nonobstant ces aléas et lacunes, la première surprise et de taille vint du niveau de retour enregistré. Il nous avait été signifié en effet qu’un taux de réponse d’environ 10% constituerait déjà une réussite, et encore à partir d’un questionnaire pas supérieur à deux pages. Les dirigeants cynégétiques estimaient franchement que la distance présumée de ces chasseurs d’avec l’écrit d’une part, le « ras-le-bol » général qui semblait chez eux omniprésent d’autre part, invalidaient passablement les chances de réussite. Il n’en fut rien, au contraire, puisque le questionnaire de quatre pages, pour 63 questions, analysées par 90 variables, a dépassé de loin toutes les attentes en matière de retour. L’ensemble de ces éléments nous permet d’estimer que les résultats de cette enquête méritent une crédibilité certaine974. Taux de participation ACDPM : 62,74% (hors retours à l’expéditeur : 19), soit 490 retours exploitables. Taux de participation PIC.02 : 31,29% (hors retours à l’expéditeur : 6), soit 51 retours exploitables. Si on les compare au taux moyen de réponse pour les enquêtes par questionnaire auto administré, lequel s’établit en France autour de 7% d’après les relevés établis, on peut estimer exceptionnelle la réception de ces études (57%). L’explication tient probablement dans l’approche adoptée, le courrier d’accompagnement étant signé du Président de la FDC.80, Yves Butel. Pourtant, l’hypothèse selon laquelle le contexte aurait joué favorablement en faveur de ce taux de réponse massif doit s’imposer comme une évidence, les dernières polémiques sur la chasse, et en particulier les débats contradictoires sur le devenir des sauvaginiers, ayant repris de plus belle dans la période pré-électorale. Plus encore pour le PIC.01, la candidature de Jean Saint-Josse à l’élection présidentielle et l’annonce des 974 Afin de favoriser la participation à l’enquête, et ce en fonction de l’association d’adhésion des publics visés, le logo correspondant a été reproduit sur la première page du questionnaire. Ainsi, chaque destinataire pouvait visuellement identifier l’organisme demandeur de l’enquête (la FDC.80) et vérifier le soutien de son association qui accordait l’emploi de son emblème. Cette précaution nous était suggérée par la méfiance, vérifiée sur d’autres terrains et à l’occasion d’autres études, dont les chasseurs font preuve à l’égard du regain d’intérêt dont ils sont l’objet désormais de la part du reste de la société, laquelle est ressentie par eux comme de plus en plus hostile à leur pratique. Par ailleurs, un encart fut inséré dans le mensuel de la Fédération des Chasseurs de la Somme (« Picardie Chasse »), et celui des chasseurs de gibier d’eau (« La Sauvagine »), invitant retourner le questionnaire. Enfin, les administrateurs des trois ACDPM ont répondu dans le même sens aux interrogations de leurs adhérents. Il est à ce propos intéressant de noter que plusieurs chasseurs de la Baie de Somme se sont rendus à leur siège social afin de « se faire aider » dans la rédaction de leurs réponses, preuve s’il en était besoin de l’extrême inquiétude quant à l’usage qu’il pourrait en être fait. Nous avons toutes les raisons de penser qu’ils ont été effectivement déboutés quant à leur demande, comme cela nous a été rapporté. De plus, la rapidité exigée pour le retour du questionnaire permettait mal les concertations. Or, la plupart des documents sont revenus dans un temps particulièrement court. 623 candidats CPNT aux législatives qui devaient suivre, ne pouvaient manquer de mobiliser les chasseurs du DPM picard. Considérés par ACDPM, ceux-ci répondent dans une proportion graduée par ce que l’on sait maintenant de leur proximité au terrain, avec pour extrême d’un côté le Littoral Sud (69,56%), désormais très majoritairement pratiqué par des locaux du Vimeu, et de l’autre la Baie d’Authie (61,29%), plus largement occupée par une population allogène, en particulier en provenance du Pas-de-Calais. 2.) Taux de réponse par type de question : un investissement qui s’exprime d’autant mieux avec les éléments biographiques Si donc la participation au questionnaire fut relativement massive, force est également de constater que ce fut le cas pour la plupart des questions posées975. Ces questions pouvaient avoir des statuts différents : les unes concernant l’intégralité de l’échantillon, les autres une part seulement, comme celles attendant une précision par rapport à un type de réponse à une interrogation précédente. Pour le PIC.01, sur les 64 questions du premier type, le plus significatif en terme de taux de réponse, la moyenne s’établit à 93,6%, allant de 59% pour les remarques, questions et ajouts personnels (espace libre à la fin du questionnaire), jusqu’à 100% en ce qui concerne les initiateurs ayant contribué à l’apprentissage de la chasse. L’investissement des répondants peut ainsi être qualifié de très fort. Les interrogations qui ont semblé générer le plus de difficultés, et donc susceptibles d’avoir été problématiques, relèvent d’une part des sujets d’actualité vecteurs de polémiques et de discussions au sein même du monde cynégétique et d’autre part des opinions et de leur justification. Pour exemples : 1.) les raisons venant en appui de leur opinion actuelle quant à Natura 2000 (64,1% de taux de réponse) 2.) les motifs expliquant l’opinion vis-à-vis des éventuelles perturbations provoquées par les activités humaines (70,7%) 3.) trois mots définissant le ressenti par rapport au littoral picard (76,9%) 4.) l’explication de l’opinion quant au devenir de la chasse (82%), 5.) les publics usagers du littoral qui pourraient gêner leur activité (85,7%). Par catégorie de question maintenant, la formulation peut appeler à la rédaction d’un texte, à un positionnement selon des modalités de réponse pré-formatées, à un nombre, une date etc. 1.) Questions fermées (modalités proposées : 96,0% de taux de réponse) 2.) questions ouvertes (texte : 85,4%) 3.) semi-ouvertes (modalités plus « autre » libre : 96,5%) 4.) questions numériques : 97,9% 5.) questions dates : 99,4%. Particulièrement révélatrices de la place occupée par la chasse dans le vécu des répondants, les dates obtiennent un score rarement égalé dans ce genre d’enquête. Exemple parmi les exemples, l’année du premier permis de chasser ne s’oublie pas facilement et semble constituer un véritable rite de passage, surtout lorsque le sésame fut décrocher au plus tôt, soit seize ans. En seconde position, les chiffres, ramenant souvent ici à des nombres d’années de pratique, sont également parfaitement maîtrisés. Seules les questions ouvertes restent plus souvent sans 975 Le premier questionnaire (cf. annexes) était composé de 43 questions fermées appelant à se ranger dans une ou plusieurs modalités de réponses pré-déterminées, 35 questions ouvertes amenant à produire un texte, 10 questions numériques impliquant des données chiffrées et 2 questions « dates ». Néanmoins, l’aspect « fermé » du questionnaire doit être nuancé par le fait que 20 questions ouvertes visaient en fait à offrir la possibilité de nuancer 19 des questions fermées. Il en était de même pour une des questions numériques. De plus, ces variables sont le produit d’un recodage, certaines questions ayant amené à des compléments d’information non sollicités que nous avons jugé utile de saisir. Pour les chasseurs de Rue, la seconde vague de questionnaires s’est enrich de l’expérience des premiers retours et de la nécessité d’interroger sur les éventuelles formes de chasse pratiquées sur le DPM le cas échéant, et ce dans un objectif comparatif, ce qui explique qu’on ait eu recours à 126 variables. 624 réponse, mais avec néanmoins un bon taux de participation. C’est par ailleurs souvent ce type de questions qui laisse le plus d’espace blanc. Antérieurement à ce travail, une première étude976 s’était attachée à interroger les visiteurs du littoral picard, quant à la manière dont ils recevaient le réseau Natura 2000 localement. L’auteur du rapport constatait dans un premier temps qu’ils étaient ainsi globalement 23,8% à déclarer avoir « entendu parler de la directive européenne », mais ce taux s’élevait jusqu’à 43,8% si l’on considérait les seuls locaux. Au total, la désapprobation touchait 37,1% de l’échantillon, dont 13,7% au nom du fait que la mise en place du réseau serait allée « à l’encontre de leurs activités de chasse et de pêche ». Pour ce qui nous concerne, l’optique visait d’emblée à répondre à ce qui, en arrière plan d’une désapprobation connue et présentée comme la proue du navire ANCGE, relevait de l’amirauté en vue sur le pont, par rapport à la réalité des cales. Ramait-on par exemple dans le même sens au plus près des flots, mais bien plus encore qui étaient les mutins en conflit avec le gros de la flotte ; comment avaient-ils réussi à modifier un temps le cap de l’armada cynégétique pour entrer en guerre sur l’océan politique. S’agissait-il, comme on l’avançait souvent dans les médias, avec parfois de troublants échos de la recherche en sciences sociales, d’une stratégie contestataire à l’allure gaullienne visant à instrumentaliser une base prolétarienne pour éperonner un lobbying plus classique en perte de vitesse? Même à imaginer les leaders du coup d’Etat en seconds couteaux désireux de jeter l’ancre dans les instances de décision pour s’amarrer à quelques strapontins électoraux, fallait-il penser les porteurs de bulletin s comme de dociles sherpas, naïvement disposés à servir de galériens? Et en premier lieu au nom de quelles valeurs? C’est ce à quoi se sont attachées les enquêtes, tant qualitatives que quantitatives, mises en place auprès de ces quatre équipages d’attaque que sont les trois ACDPM du littoral picard, épaulées de ce bâtiment en fond de rade qu’est l’ACGECR, étroitement imbriquée avec la SCCR.	9,445
https://openalex.org/W4388491428_2	Spanish-Science-Pile	Open Science	Various open science	2,023	REENCUENTROS CON TINTES DRAMÁTICOS TRAS LA GRAN RECESIÓN. LES DISTÀNCIES (2018) DE ELENA TRAPERO	None	Spanish	Spoken	2,852	5,706	Dejando a un lado el contexto económico que el filme muestra, otras causas (más universales para cualquier generación) conllevan la desintegración de dicho grupo de amigos. En primer lugar, la desaparición de sus puntos en común, la bifurcación de los caminos que cada uno toma, la distancia física y emocional que se alza entre ellos. Dicen que los amigos se eligen y la familia no, aunque al cabo de unos años puede ser que no nos despeguemos de ciertas/os amigas/os por costumbre, por miedo al aislamiento, y no por elección. Y en el filme, estas diferencias son palpables en todos los diálogos. Solo hay un destello de luz y esperanza para su relación, solo una única cosa los une sin complejos: los recuerdos de los viejos tiempos. Pero, aunque por unos instantes hablen el mismo lenguaje, ese momento es fugaz, no es suficiente para cohesionarlos y salvar su amistad. En segundo lugar, la transición de la juventud a la vida adulta puede ser complicada, y unas personas pueden aferrarse demasiado a ciertas épocas de la vida, como la etapa universitaria. Acaba llegando el momento en el cual aceptamos que esas noches y fiestas de la juventud se han esfumado, como le pasa a Eloy en la escena del bar del inicio de la película. Además, a los veinte años hay espacio para la esperanza y la reinvención, pero a medida que pasa el tiempo, esa sensación va dejando paso a la frustración. Al ser preguntados en el coloquio por sus conclusiones respecto al largometraje, Esparbé, Sevilla y Trapé coinciden en que la mayor lección que sacan es la de aprender a dejar ir, aunque sea lo que más duele, porque ·crecer es también saber decir adiós. Eso no nos hace mejores ni peores, simplemente es lo que toca, es lo responsable. Esas épocas de cambio pueden ser muy dolorosas, ya que FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 325 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero implican desprendernos de una parte de nosotros, pero son también muy enriquecedoras. Además de ese minucioso desplome de la amistad, la directora abarca otros ámbitos que le interesan, siempre bajo ese sombrío Berlín (toda la película se rodó allí para favorecer la sensación de frialdad), con sus tonos grises y apagados, que van oscureciéndose a medida que transcurre el argumento. Ya había indagado en su ópera prima Blog (2010) el cambio de paradigma que suponen las nuevas tecnologías y sus efectos a la hora de relacionarnos. Esta vez explora el papel (inútil) de las llamadas y los mensajes de voz en ciertas situaciones críticas e inesperadas que pueden suceder. En segundo lugar, el embarazo del personaje de Olivia juega también un papel relevante, sobre el cual se exploran las obligaciones y prohibiciones que acarrea. Tal y como la directora expone, quería tratar la maternidad, que se presupone que es un estado de bendición de la mujer, un instinto que no se cuestiona, y que muchas mujeres viven y sufren. Algo tan absurdo como el no poder dejar de fumar se convierte en una cosa que se hace mal y por la cual se culpabiliza a la mujer. Olivia está empeñada en parar el tiempo, porque cuando tenga el hijo ya todo va a cambiar. Por último, el declive de la amistad se vincula al declive de una pareja que se encuentra en un punto de no retorno de la relación, en la cual uno de los miembros se desencanta del otro al ver cómo trata de forma hostil a sus amigos. Imagen 10. Fotograma de Les distàncies (2018). Álex (Miki Esparbé) en el metro de Berlín. Fuente: www.filmaffinity.com/es/film108072.html. En 2005, el dúo Amaral publicaba la canción Marta, Sebas, Guille, y los demás con ciertas similitudes con el filme que, de manera más amable, decía FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 326 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero Carlos me contó que a su hermana Isabel la echaron del trabajo, sin saber por qué, no le dieron ni las gracias porque estaba sin contrato, aquella misma tarde fuimos a celebrarlo […] ¿Dónde empieza y dónde acabará el destino que nos une y que nos separará? […] son mis amigos, en la calle pasábamos las horas, son mis amigos por encima de todas las cosas […]. Alicia fue a vivir a Barcelona, y hoy ha venido a la memoria, Claudia tuvo un hijo, y de Guille y los demás ya no sé nada. Se retrata, así, el luto que suponen las rupturas de amistad, a veces eclipsado por las rupturas de pareja o los fallecimientos de familiares. Es, sin embargo, un luto que puede llegar a ser igual de doloroso, ya que las personas más cercanas en la juventud pueden convertirse, de un día para otro, en desconocidos, tal y como se muestra en una escena en la cual Guille y Álex se encuentran en un bar, sin nada que decirse. Quedará en nuestra mente otro episodio de la película: una de las miradas más tristes que se pueden recordar en el cine reciente. Al final del largometraje Eloy y Álex se encuentran en el metro, estando cada uno en una vía: se atisban, comprenden en ese momento todo lo que ha sucedido, entienden que no podrán volver atrás, se vuelven conscientes de su propia soledad y aceptan su propia desgracia, que se ha manifestado de forma inesperada y cruel ese fin de semana que prometía ser memorable, pero que acaba siendo el más funesto y compungido que podrán recordar. REFERENCIAS BIBLIOGRÁFICAS Alberoni, Francesco (2006). La amistad. Aproximación a uno de los más antiguos vínculos humanos. Barcelona: Gedisa. Asión, Ana (2018). El cambio ya está aquí. 50 películas para entender la Transición española. Barcelona: UOC. Beck, Ulrich (2007). Un nuevo mundo feliz. La precariedad del trabajo en la era de la globalización. Barcelona: Paidós Ibérica. Berardi, Franco (2007). El sabio, el mercader y el guerrero. Del rechazo del trabajo al surgimiento del cognitariado. Madrid: Acuarela. Caparrós Lera, José María (2005). La Pantalla Popular. El cine español durante el Gobierno de la derecha (1996-2003). Madrid: Akal. ___ (2017). El cine español durante el Gobierno de Zapatero (2004-2011). Madrid: Biblioteca Nueva. Cucó Giner, Josepa (1995). La amistad: una perspectiva antropológica. Barcelona: Icaria/ Institut Català d’Antropologia. FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 327 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero Homans, George, C. (1963). El grupo humano. Buenos Aires: Eudeba. Muir, Edward (1994). Fiesta y rito en la Europa moderna. Madrid: Complutense. Requena Santos, Félix (1994). Amigos y redes sociales. Elementos para una sociología de la amistad. Madrid: CIS/Siglo XXI. Riechmann, Jorge, Fernández Buey, Francisco (1995). Redes que dan libertad. Introducción a los nuevos movimientos sociales. Barcelona: Paidós Ibérica. Sánchez Barba, Francesc (2019). Visiones de la amistad. El cine del reencuentro. Canterano (Roma): Aracne. Schult, Uwe (1993). La fiesta. Una historia cultural desde la Antigüedad hasta nuestros días. Madrid: Alianza Editorial. Sennett, Richard (2006). La corrosión del carácter. Las consecuencias personales del trabajo en el nuevo capitalismo. Barcelona: Anagrama. Sorlin, Pierre (1983). Sociología del cine. La apertura para la historia del mañana. México D.F.: Fondo de Cultura Económica. Van Dam, Andrew. “The unluckiest generation in U.S. history”. The Washington Post, 5 de Jun. 2020. www.washingtonpost.com/business/2020/05/27/millennialrecession-covid/ FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 328 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero ANEXO. PELÍCULAS SOBRE LA AMISTAD EN EL CINE ESPAÑOL (SE INCLUYEN LAS COPRODUCCIONES) DURANTE EL SIGLO XXI 3 2000 (13 de 98 títulos). Ataque verbal de Miguel Albaladejo, Besos para todos, de Jaime Chávarri, El cielo abierto, de M. Albaladejo, En construcción, de J.L. Guerín, Érase otra vez, de Juan Pinzás, Km. 0, de J.L. Iborra & Y. García, Krámpack, de Cesc Gay, Lo mejor de cada casa (una semana en el parque), de Toni Abad, Maestros, de Óscar del Caz, Noche de fiesta, de Xavi Puebla, Nosotras, de Judith Colell, Las razones de mis amigos, de Gerardo Herrero, Todo me pasa a mí, de Miquel García. 2001 (7 de 106) Amigorama, de J.M. Nunes, Antigua vida mía de Héctor Olivera, Canícula, de A. García-Capelo, Clara & Elena, de M. Iborra, Gente pez, de Jorge Iglesias, El paraíso ya no es lo que era, de Francesc Betriu, Pau i el seu germà, de Marc Recha. 2002 (8 de 136) Amnesia, de G. Salvatore, Hable con ella, de P. Almodóvar, Los lunes al sol, de F. León de Aranoa, Piedras, de R. Salazar, Planta 4ª, de A. Mercero, El sueño de Ibiza, de Igor Fioravanti, La tarara del chapeo, de Enrique Navarro, Una casa de locos, de Cédric Klapisch. 2003 (4 de 109) En la ciudad, de Cesc Gay, La fiesta, de M. Sanabria & C. Villaverde, Noviembre, de Achero Mañas, Viaje a Narragonia, de Germán Berger. 2004 (4 de 134) Mundo fantástico, de Max Lemcke, El abrazo partido, de Daniel Burman, El milagro de Candeal, de Fdo. Trueba, Luna de Avellaneda, de J.J. Campanella. 2005 (7 de 142) 7 vírgenes, de Alberto Rodríguez, Aguaviva, de Ariadna Pujol, El calentito, de Chus Gutiérrez, Malas temporadas, de Manuel Martín, Princesas, de F. León de Aranoa, Sínfin, el retorno del rock, de C. Villaverde & M. Sanabria, Tapas, de J. Corbacho & J. Cruz. 2006 (4 de 141) Arena en los bolsillos, de César Martínez, Tu vida en 65’, de Maria Ripoll, Va a ser que nadie es perfecto, de J. Oristrell, Remake, de Roger Gual. 33 Elaboración propia a partir de: Anuarios ICAA publicados por el Ministerio de Cultura: en papel entre 2001 y 2007 y en la página web (mcu.gob.es) en los sucesivos y de la base de datos Filmaffinity (www.filmaffinity.com/es/main.html). Entre paréntesis los títulos detectados (posiblemente haya más) de la producción anual. FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 329 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero 2007 (6 de 172) Atlas de Geografía Humana, de Azucena Rodríguez, Bajo las estrellas, de Félix Viscarret, Café solo... o con ellas, de Álvaro Díez, Déjate caer, de Jesús Ponce, La torre de Suso, de Tom Fernández, Ciudad en celo, de Hernán Gaffet. 2008 (6 de 173) Gente de mala calidad, de J.J. Cavestany, Pradolongo, de Ignacio Vilar, Una palabra tuya, de A. González-Sinde, El juego del ahorcado, de M. Gomez Pereira, Mi única familia, de Miguel Óscar, Ruido, de Marcelo Bertalmio. 2009 (8 de 308) 7 Minutos, de Daniela Féjerman, After, de Alberto Rodríguez, La isla interior, de F. Ayuso & D. Sabroso, Gordos, de D. Sánchez Arévalo, Malas noticias, de M.A. Cárcano, Mentiras y gordas, de A. Albacete & D. Menkes, Tres días en familia, de Mar Coll, V.O.S., de Cesc Gay. 2010 (10 de 293) 9 meses, de M. Perelló, Blog, de Elena Trapé, Bon appetit, de David Pinellos, Cuidadores, de Óskar Tejedor, Diario de Carlota, de J.M. Carrasco, Héroes, de Pau Freixas, Pájaros de papel, de Emilio Aragón, Scouts (en la senda), de J. López & F. J. Menéndez & M.Á. Pérez, Terrados, de Demian Sabini, La vida empieza hoy, de Laura Mañá. 2011 (11 de 295) Aficionados, de Arturo Dueñas, Amigos…, de B. Manso & M. Cabota, Any de Gràcia, de Ventura Pons, Arrugas, de I. Ferreras, Cara de queso: mi primer gueto, de Ariel Winograd, Katmandú, un espejo en el cielo, de Icíar Bollaín, La montaña rusa, de E. Martínez Lázaro, La plaza, de A. Morán, Primos, de D. Sánchez Arévalo, La Trinca: biografía no autorizada, de J. Oristrell, Viene una chica, de Chema Sarmiento. 2012 (7 de 259) Onde Está a Felicidade?, de C.A. Ricelli, Dormíamos, despertamos, de A. Domingo et al, Elefante blanco, de Pablo Trapero, FIN, de J. Torregrossa, Libre te quiero, de B. Martín Patino, Los niños salvajes, de Patricia Ferreira, Zuloak, de Fermín Muguruza. 2013 (6 de 289) El árbol magnético, de Isabel de Ayguavives, La estrella, de Alberto Aranda, Máscaras, de Iago González, Viaje a Surtsey, de J. Asenjo & M.Á. Pérez, La vida inesperada, de Jorge Torregrossa, Vivir es fácil con los ojos cerrados, de David Trueba. 2014 (9 de 257) 2 francos, 40 pesetas, de Carlos Iglesias, Las altas presiones, de Ángel Santos, El club de los incomprendidos, de Carlos Sedes, Cómo sobrevivir a una despedida, de Manuel Moreno, Dos amigos de Polo Menárguez, Felices 140, de Gracia Querejeta, El futuro, FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 330 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero de L. López Carrasco, Historias de Lavapiés de Ramón Luque, Un sitio donde quedarse, de M. Arribas & A. Pérez. 2015 (11 de 296) A cambio de nada, de Daniel Guzmán, Los exiliados románticos, de Jonás Trueba, Los héroes del mal, de Zoe Berriatúa, Isla bonita, de Fdo. Colomo, Losers, de O. Pérez & S. Soler, Mi panadería en Brooklyn, de Gustavo Ron, Muchos pedazos de algo, de David Yáñez, Perdidos en el Norte, de I. García Velillo, Tres días en Pedro Bernardo, de Daniel Andrés Sánchez, Truman, de Cesc Gay, Un otoño sin Berlín, de Lara Izaguirre. 2016 (8 de 244) Antonio cumple 50, de Alejandro Ferrer, Nueve bares, de Ángel Puado, Alalá (Alegría), de Mª R. Malvárez, Bittersweet days, de Marga Melià, El olivo, de Icíar Bollaín, Las furias, de Miguel del Arco, Les amigues de l’Àgata, de Laia Alabert et al, Proyecto USA, de Miguel Herrero. 2017 (12 de 268) Casi leyendas, de Gabriel Nesci, Dream Songs, de Agustí Vilà, El mundo es suyo, de Alfonso Sánchez, Jean-François y el sentido de la vida, de Sergi Portabella, La llamada, de J. Calvo & J. Ambrossi, La última aventura del Gandul, de Tomás Cimadevilla, Les distàncies, de Elena Trapé, Sergio & Serguéi, de Ernesto Daranas, Smoking Club (129 normas), de Alberto Utrera, Sólo en un día, de R. Álvarez y J.L. Noriega, Thi Mai. Rumbo a Vietnam, de Patricia Ferreira, Terra ferma, de C. MarquésMarcet. 2018 (12 de 268) Seguidores, de Iván Fernández, 4 Latas, de Gerardo Olivares, Abuelos, de Santiago Requejo, Ardara, les passes perdudes, de R. Fransoy & X. Puig, Cabanyal, de Fréderique Pressman, Campeones, de Javier Fesser, El increíble finde menguante, de Jon Mikel Caballero, La estación violenta, de Anxos Fazáns, Nosotros, de Felipe Vara, Paella Today, de César Sabater, The Best Day Of My Life, de Fernando González Molina, Try de Ángel, de Haro. 2019 (14 de 258) ¿A quién te llevarías a una isla desierta?, de Jota Linares, Be Happy!, de Ventura Pons, Buñuel en el laberinto de las tortugas, de Salvador Simó, Érase una vez, de Primitivo Pérez, Invisibles, de Gracia Querejeta, Isaac, de David Matamoros, La banda, de Roberto Bueso, La innocència (La inocencia), de Lucia Alemany, La lista de los deseos, de Álvaro Díaz, Las niñas, de Pilar Palomero, Litus, de Dani de la Orden, Lo dejo cuando quiera, de Carlos Therón, Ojos negros, de M. Lallana, I. Castelo, I. Alarcón & S. García, Operación Globus, de Ariadna Seuba. FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 331 Arlet Sánchez Rojals y Francesc Sánchez Barba Reencuentros con tintes dramáticos tras la gran recesión. Les distàncies (2018) de Elena Trapero 2020 (7 de 222) Chavalas, de Carolina Rodríguez, Coses a fer abans de morir, de M. Llorens & C. Fernández, Descarrilados, de Fer García-Ruiz, El sitio de Otto, de Oriol Puig, La vida era eso, de David Martín, Les dues nits d’ahir, de Pau Cruanyes, Los europeos, de Víctor García León. 2021 (8 de 290) Alegría, de Violeta Salama, El club del paro, de David Marqués, Escape Room, de Héctor Claramunt, La cançó pop, de Raul Portero, La passió segons Pep Amores, de Júlia Girós, La revolución bailando, de Eli Martín, Live is Life, de Dani de la Torre, Libertad, de Clara Roquet. 2022 (10 de 343) Amigos hasta la muerte, de Javier Veiga, Buscando la película (Verano 2020), de E. García-Vázquez, De perdidos a Río, de Joaquín Mazón, Isósceles, de Ignacio Nacho, La mala familia, de Nacho A. Villar, La manzana de oro, de Jaime Chávarri, Les vacances, de Mara de Elena Escura, Nosaltres no ens matarem amb pistoles, de Maria Ripoll, Quan no acaba la nit, de Óscar Montón, Tenéis que venir a verla, de Jonás Trueba. 2023 (11 de 181 hasta el 23/09/2023) Alguien que cuide de mí, de E. Lindo & D. Féjerman, Amigos hasta la muerte, de Javier Veiga (España-México), Las buenas compañías, de Sílvia Munt, Campeonex, de Javier Fesser, Las chicas están bien, de Itsaso Arana, De perdidos al río, de Joaquín Mazón, La manzana de oro, de Jaime Chávarri, Mi soledad tiene alas, de Mario Casas, La pecera, de G. Marrero, Te estoy amando locamente, de Alejandro Marín, Viento sur, de A. García-Capelo. Total: 203 (de 5282 títulos). FILMHISTORIA Online Vol. 33, núm. 1 (2023)· ISSN: 2014-668X 332.	2,643
https://openalex.org/W4283760617	OpenAlex	Open Science	CC-By	2,022	PSOAS ABSCESS:A RARE LOCALIZATION OF TUBERCULOSIS INFECTION	Lotfi Azzouz	English	Spoken	3,277	5,815	Introduction:- Tuberculosis is an infectious disease that can affect several organs and systems. It can involve almost the whole body, including the abdomen. Retroperitoneal involvement is rare. Extra-pulmonary tuberculosis remains a difficult diagnosis to make, because of its often non-specific manifestations. However, since there are no pathognomonic imaging findings, the diagnosis ultimately relies on histopathological and microbiological confirmation. Before the discovery of modern tuberculosis treatment, psoas abscess was a well-known complication of tuberculosis of the spine, where it was first described by Mynter in 1881, who called it psoitis. [1] We report a rare case of retroperitoneal abscess of the psoas whose ISSN: 2320-5407 ISSN: 2320-5407 Int. J. Adv. Res. 10(06), 23-28 Journal Homepage: -www.journalijar.com Article DOI:10.21474/IJAR01/14852 DOI URL: http://dx.doi.org/10.21474/IJAR01/14852 PSOAS ABSCESS:A RARE LOCALIZATION OF TUBERCULOSIS INFECTION Lotfi Azzouz, Fatima Zohra Benmoula, Laila Dahbi-Skali, Marouane Baiss, Mdaghri Jalil, Benamar Said, Mssrouri Rahal and Abdellatif Settaf …………………………… Manuscript Info ……………………. Manuscript History Received: 05 April 2022 Final Accepted: 08 May 2022 Published: June 2022 Key words:- Psoas Abscess, Tuberculosis ……………………………………………………………… Psoas abscess is a rare disease with various etiologies. The most common pathogen in primary abscess is Staphylo coccus aureus. Crohn's disease is the most common cause of secondary abscess. Mycobacterium tuberculosis is considered an extremely rare cause of psoas abscess, The clinical manifestation is insidious, the classic symptoms are pain and fever. Computed tomography (CT) is the gold standard in the diagnosis and follow-up of retroperitoneal abscesses. The treatment of choice is percutaneous drainage guided by imagery. Surgical drainage should be reserved for cases of failure of percutaneous drainage, if possible via the extraperitoneal route, but in the event of a secondary abscess, the transperitoneal approach is preferred to correct the original disease. Copy Right, IJAR, 2022,. All rights reserved. Copy Right, IJAR, 2022,. All rights reserved. …………………………………………………………………………………………………….... Introduction:- Tuberculosis is an infectious disease that can affect several organs and systems. It can involve almost the whole Observation:- A 44-year-old woman was admitted to our surgery department for left cervical swelling. In these antecedents we find the notion of pulmonary tuberculosis treated in childhood. She had no surgical history. His history of the disease goes back to 2 months of his admission by the appearance of a left laterocervical lymphadenopathy increasing in size gradually with notion of dysphagia having motivated a consultation with a general practitioner. An abdominal ultrasound was ordered and revealed the presence of a cystic mass in the left iliac fossa measuring 13 × 6 cm, probably related to a left ovarian cyst. Furthermore, the patient presented no associated signs, and all progressed in a context of conservation of general condition and apyrexia. The clinical examination on admission found the patient to be hemodynamically and respiratory stable, with a temperature of 37°C, a heart rate of 75/min and blood pressure of 130/60 mmHg. The cervical examination objectified the presence of left laterocervicaladenopathies, on the other hand the abdominal examination was unremarkable. The biological assessment was normal with hemoglobin at 12.1 g/dl, white blood cells at 7610 /µl, urea at 0.18g/l, creatinine at 6.6mg/l and a CRP at 5.79mg/l. during the hospitalization an abdominal and cervical ultrasound was redone and which showed at the cervical level, 23 Corresponding Author:- Lotfi Azzouz Corresponding Author:- Lotfi Azzouz Int. J. Adv. Res. 10(06), 23-28 ISSN: 2320-5407 ISSN: 2320-5407 the presence at the left lateral level of an anechoic, heterogeneous, well-limited oval formation, regular contours, coming into contact with the jugular vein internal left, not taking the color Doppler measuring 15 × 38 × 36 mm in favor of left necrotic ADP. At the abdominal level, the presence of a cystic mass in the left iliac fossa measuring 136 × 61 mm very probably related to a left ovarian cyst. The radiological assessment was completed by a CERVICO- THORACO-ABDOMINO-PELVIC CT scan, the result of which was in favor of a necrotic ADP of the left IV chain, measuring: 42x19mm with the presence of a collection of the left psoas muscle, well limited, hypodense, enhanced at the periphery after injection of PDC, measuring: 61x66x107mm (APXT×H). And a left laterouterine mass, of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. Observation:- (Figure 1) the presence of a cystic mass in the left iliac fossa measuring 136 × 61 mm most likely related to a left ovarian cyst. The radiological assessment was completed by a CERVICO-THORACO-ABDOMINO-PELVIC CT scan, the result of which was in favor of a necrotic ADP of the left IV chain, measuring: 42x19mm with the presence of a collection of the left psoas muscle, well limited, hypodense, enhanced at the periphery after injection of PDC, measuring: 61x66x107mm (APXT×H). And a left laterouterine mass, of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. (Figure 1) the presence of a cystic mass in the left iliac fossa measuring 136 × 61 mm most likely related to a left ovarian cyst. The radiological assessment was completed by a CERVICO-THORACO-ABDOMINO-PELVIC CT scan, the result of which was in favor of a necrotic ADP of the left IV chain, measuring: 42x19mm with the presence of a collection of the left psoas muscle, well limited, hypodense, enhanced at the periphery after injection of PDC, measuring: 61x66x107mm (APXT×H). And a left laterouterine mass, of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. (Figure 1) The radiological assessment was completed by a CERVICO-THORACO- ABDOMINO-PELVIC CT scan, the result of which was in favor of a necrotic ADP of the left IV chain, measuring: 42x19mm with the presence of a collection of the left psoas muscle, well limited, hypodense, enhanced at the periphery after injection of PDC, measuring: 61x66x107mm (APXT×H). And a left laterouterine mass, of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. (Figure 1) The radiological assessment was completed by a CERVICO-THORACO-ABDOMINO-PELVIC CT scan, the result of which was in favor of a necrotic ADP of the left IV chain, measuring: 42x19mm with the presence of a collection of the left psoas muscle, well limited, hypodense, enhanced at the periphery after injection of PDC, measuring: 61x66x107mm (APXT×H). And a left laterouterine mass, of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. (Figure 1) of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. (Figure 1) of pure fluid density, not enhanced after injection of PDC, measuring: 30x32mm, related to an ovarian cyst. Observation:- (Figure 1) Figure 1:- CT image showing a left psoas abscess (arrow). Figure 1:- CT image showing a left psoas abscess (arrow). 24 Int. J. Adv. Res. 10(06), 23-28 The was positive for Mycobacterium tuberculosis. According to these results, the patient treatment with isoniazid, rifampicin, pyrazinamide and ethambutol for 2 months. This by isoniazid and rifampicin for another four months without reporting any side effects. No inistered with a good clinical and radiological evolution on a control abdominal CT scan 6 Figure 4:- Purulent appearance of drainage fluid. Figure 5:- Drainage of a left cervical abscess. Figure 4:- Purulent appearance of drainage fluid. Figure 4:- Purulent appearance of drainage fluid. Figure 4:- Purulent appearance of drainage fluid. Figure 5:- Drainage of a left cervical abscess. Figure 5:- Drainage of a left cervical abscess. Figure 5:- Drainage of a left cervical abscess. Histological examination was compatible with the presence of granulomas with caseous necrosis. The microbiological culture was positive for Mycobacterium tuberculosis. According to these results, the patient received antituberculosis treatment with isoniazid, rifampicin, pyrazinamide and ethambutol for 2 months. This treatment was followed by isoniazid and rifampicin for another four months without reporting any side effects. No other antibiotic was administered with a good clinical and radiological evolution on a control abdominal CT scan 6 months later. Histological examination was compatible with the presence of granulomas with caseous necrosis. The microbiological culture was positive for Mycobacterium tuberculosis. According to these results, the patient received antituberculosis treatment with isoniazid, rifampicin, pyrazinamide and ethambutol for 2 months. This treatment was followed by isoniazid and rifampicin for another four months without reporting any side effects. No other antibiotic was administered with a good clinical and radiological evolution on a control abdominal CT scan 6 months later. Int. J. Adv. Res. 10(06), 23-28 Percutaneous aspiration of the mass, under ultrasound control, gave a mucinous liquid but insufficient for mycobacteriological examination. For this reason the indication of retroperitoneal surgical drainage was raised. The patient underwent surgery with extraperitoneal drainage through a Jalaguier incision in the left iliac fossa (Figure 2.3) obtaining 300 ml of purulent material from the psoas. (Figure 4) obtaining 300 ml of purulent material from the psoas. (Figure 4) Figure 2:- Retroperitoneal abscess that gives a bulging appearance of the posterior parietal peritoneum. Figure 2:- Retroperitoneal abscess that gives a bulging appearance of the posterior parietal peritoneum. Picture 3:- Extraperitoneal drainage. Picture 3:- Extraperitoneal drainage. At the cervical level: a left cervical incision along the internal edge of the sternocleidomastoid muscle was performed with the discovery of a retro muscular abscess which was punctured and drained by a Redon drain. (Figure 5) The postoperative evolution of the patient was good. Discharge on D3 postoperative. Picture 3:- Extraperitoneal drainage. At the cervical level: a left cervical incision along the internal edge of the sternocleidomastoid muscle was performed with the discovery of a retro muscular abscess which was punctured and drained by a Redon drain. (Figure 5) The postoperative evolution of the patient was good. Discharge on D3 postoperative. At the cervical level: a left cervical incision along the internal edge of the sternocleidomastoid muscle was performed with the discovery of a retro muscular abscess which was punctured and drained by a Redon drain. (Figure 5) The postoperative evolution of the patient was good. Discharge on D3 postoperative. 25 ISSN: 2320-5407 Int. J. Adv. Res. 10(06), 23-28 Int. J. Adv. Res. 10(06), 23-28 ISSN: 2320-5407 ISSN: 2320-5407 Figure 4:- Purulent appearance of drainage fluid. Figure 5:- Drainage of a left cervical abscess. Histological examination was compatible with the presence of granulomas with caseo microbiological culture was positive for Mycobacterium tuberculosis. According to these received antituberculosis treatment with isoniazid, rifampicin, pyrazinamide and ethambutol treatment was followed by isoniazid and rifampicin for another four months without reporting a other antibiotic was administered with a good clinical and radiological evolution on a control ab months later. Discussion:- PR i f ti i i t d ith d l d di i d hi h t lit t i f Figure 4:- Purulent appearance of drainage fluid. Figure 5:- Drainage of a left cervical abscess. n was compatible with the presence of granulomas with caseous necrosis. ISSN: 2320-5407 ISSN: 2320-5407 Mycobacterium tuberculosis was the cause responsible for most cases of psoas abscess. As the TB pathogen was brought under control, secondary abscesses became more common. [4] Currently, the most frequent cause of primary abscesses is Staphylococcus aureus in 88.4%, followed by streptococcus and Escherichia coli [5.6]. Crohn's disease is the most common cause of secondary abscesses. Abdominal tuberculosis is rare and is usually found in patients with severe disseminated disease [7]. Tuberculosis more frequently affects immunocompromised patients, such as HIV-infected subjects or patients with diabetes mellitus and end-stage renal disease. [8] Our country is considered an endemic area, so reactivation of a latent infection should be considered possible, even in an immunocompetent subject. In our patient, the retroperitoneal localization of tuberculosis was associated with cervical lymph node localization. There is no associated pulmonary or vertebral involvement. Immunosuppression was ruled out by negative routine blood tests, normal immunoglobulin levels, and negative HIV tests. No other bacterial infection was found in blood culture and the Percutaneous aspiration of fluid was insufficient for mycobacteriological examination. Ultrasound is a widely available method and therefore usually the primary method for detecting an abscess, but computed tomography (CT) of the abdomen remains the gold standard in the diagnosis and follow-up of retroperitoneal abscesses. [9] The diagnostic sensitivity of ultrasound for retroperitoneal abscesses is 67% to 87%, while that of CT is 90% to 100%. Tuberculosis can be detected by chest X-ray (33%) and abdominal computed tomography (88%), which are the most frequently used imaging modalities, but there were no pathogenetic criteria. [10,11]. Thus, there are no techniques for specific imaging for retroperitoneal abscesses secondary to tuberculosis infection. In our context, the chest X-ray was normal and it was the CT scan that revealed the presence of an abscess of the left psoas muscle. Treatment of retroperitoneal abscesses can be conservative with antibiotic therapy, interventional with percutaneous drainage of the abscess, and surgical with evacuation of the abscess. In many case series analyses, interventional radiation therapy with antibiotics was the recommended treatment modality. However, no randomized clinical trials have been reported on the treatment of retroperitoneal abscesses. Ultrasound-guided drainage is the most common interventional treatment. Surgery was the main treatment method along with antibiotic treatment in Huang et al. [12, 13] Drainage by puncture can be technically difficult in some patients due to the location of the abscess or its multilocular appearance. Conclusion:- Tuberculous psoas abscess is a pathology that mainly affects immunocompromised patients. On the other hand, the tuberculous etiology of a retroperitoneal abscess should be kept in mind in an immunocompetent patient if he comes from endemic areas. Management is essentially based on drainage of the abscess, if possible by puncture under radiological control, if not surgical drainage remains the definitive treatment in association with antituberculosis therapy. ISSN: 2320-5407 Surgery is reserved if percutaneous drainage fails [14]. The preferred surgical drainage is usually extraperitoneally, as we did in this case. However, patients with secondary abscesses require correction of the underlying pathology, in addition to adequate drainage. In a few reported cases of retroperitoneal abscess with tuberculosis infection, the diagnosis was usually confirmed by surgery. [15, 16]. L' pathological study of the surgical specimens revealed a granuloma and giant cells with necrotic tissue. The culture of the samples was positive. A positive culture can be found when the histological examination is negative. [17] Our patient underwent ultrasound-guided drainage, which was insufficient, hence the need to supplement with extraperitoneal surgical drainage, the anapath of the surgical specimens of which was suggestive of tuberculosis infection by the presence of granulomas with caseous necrosis. The microbiological culture was positive for Mycobacterium tuberculosis. Standard four-drug anti-tuberculosis therapy is the mainstay of management of abdominal tuberculosis for at least 6 months [10]. Our patient received anti-tuberculosis treatment for 6 months, 2. Harris LF, Sparks JE. Retroperitoneal abscess. Case report and literature review. Dig Dis Sci 1980 ;25(5):392-5. 3. Crepps JT, Welch JP, Orlando R 3rd. Management and evolution of retroperitoneal abscesses. Ann Surg 1987 ;205(3) :276-81. g ; 5. Agrawal S, Dwivedo A, Khan M. Primary psoas abscess. Dig Dis Sci 2002 ;47 :2103-2105. Dwivedo A, Khan M. Primary psoas abscess. Dig Dis Sci 2002 ;47 :2103-2105. 1. Agrawal S, Dwivedo A, Khan M. Primary psoas abscess. Dig Dis Sci 2002 ;47 :2103-2105. 2. Harris LF, Sparks JE. Retroperitoneal abscess. Case report and literature review. Dig Dis Sci 1980 ;25(5):392-5. 3. Crepps JT, Welch JP, Orlando R 3rd. Management and evolution of retroperitoneal abscesses. Ann Surg 1987 ;205(3) :276-81. 4 Santaella R Fishman E Lipsett P Primary versus secondary iliopsoas abscess : presentation microbiology and ( ) 4. Santaella R, Fishman E, Lipsett P. Primary versus secondary iliopsoas abscess : presentation, microbiology, and treatment. Arch Surg 1995 ;130 :1309-1313. Discussion:- PR space infection is associated with delayed diagnosis and a high mortality rate, ranging from 22 to 46% [2]. Typical symptoms include non-specific and non-localized abdominal pain, variable gastrointestinal upset, and generalized symptoms, such as chills, sweating, fever, or malaise. About 10% of patients are asymptomatic [3]. Our patient reported no abdominal pain and no abnormalities or signs of peritonitis were found on abdominal examination. The only symptom was a history of progressive cervical adenopathy. Psoas abscess was first described by Mynter in 1881. It is relatively rare in Western countries. It can be primary due to infections from unknown foci that spread hematogenously, or secondary to retroperitoneal or intra-abdominal abscesses, associated with another cause that facilitates its development, for example Crohn's disease. At the beginning of the last century 26 Int. J. Adv. Res. 10(06), 23-28 ISSN: 2320-5407 The References:- 1. Agrawal S, Dwivedo A, Khan M. Primary psoas abscess. Dig Dis Sci 2002 ;47 :2103-2105. 2. Harris LF, Sparks JE. Retroperitoneal abscess. Case report and literature review. Dig Dis Sci 1980 ;25(5):392-5. 3. Crepps JT, Welch JP, Orlando R 3rd. Management and evolution of retroperitoneal abscesses. Ann Surg 1987 ;205(3) :276-81. 4. Santaella R, Fishman E, Lipsett P. Primary versus secondary iliopsoas abscess : presentation, microbiology, and treatment. Arch Surg 1995 ;130 :1309-1313. 5. Agrawal S, Dwivedo A, Khan M. Primary psoas abscess. Dig Dis Sci 2002 ;47 :2103-2105. 6. Ricci M, Rose F, Meyer K. Pyogenic psoas abscess : global variations in etiology. World J Surg 1968 ;64 :834- 843. The References:- 3. Crepps JT, Welch JP, Orlando R 3rd. Management and evolution of retroperitoneal abscesses. Ann Surg 1987 ;205(3) :276-81. 4. Santaella R, Fishman E, Lipsett P. Primary versus secondary iliopsoas abscess : presentation, microbiology, and treatment. Arch Surg 1995 ;130 :1309-1313. 6. Ricci M, Rose F, Meyer K. Pyogenic psoas abscess : global variations in etiology. World J Surg 1968 ;64 :834- 843. 27 ISSN: 2320-5407 Int. J. Adv. Res. 10(06), 23-28 Int. J. Adv. Res. 10(06), 23-28 7. SK Sharma, D. Smith-Rohrberg, M. Tahir, A. Mohan, and A. Seith, “Radiological manifestations of splenic tuberculosis : a case series of 23 patients from India,” Indian Journal of Medical Research, vol. 125, no. 5, pages 669–678, 2007 p g 8. V. Nissapatorn, I. Kuppusamy, FP Josephine, I. Jamaiah, M. Rohela, and A. Khairul Anuar, “Tuberculosis : A resurgent disease in immunocompromised patients,” The South Asian Journal of Tropical Medicine and Public Health Southeast, vol. 37, supplement 13, p. 153–160, 2006. , , pp , p , 9. Kawashima A, Sandler CM, Goldman SMetal (1997) CT of inflammatory kidney disease. Radiographs 17(4):851-866 10. I. Hassan, ES Brilakis, RL Thompson, and FG Que, “Surgical management of abdominal tuberculosis,” Journal of Gastrointestinal Surgery, vol. 6, no. 6, pp. 862–867, 2002. g y pp 11. A. Uzunkoy, M. Harma, and M. Harma, “Diagnosis of abdominal tuberculosis : experience of 11 cases and review of the literature,” World Journal of Gastroenterology, vol. 10, no. 24, pp. 3647–3649, 2004. 12. RF Coelho, ED Schneider-Monteiro, JLB Mesquita, E. Mazzucchi, A. MarmoLucon and M. Srougi, “Renal and perinephretic abscesses: analysis of 65 consecutive cases”, World Journal of Surgery, vol. 31, no. 2, p. 431–436, 2007. 13. AR EL-Nahas, R. Faisal, T. Mohsen, MS AL-Marhoon, and H. Abol-Enein, “What is the best drainage method for a perinephric abscess ?” International Brazilian Journal of Urology, vol. 36, no. 1, p. 29–37, 2010 g for a perinephric abscess ?” International Brazilian Journal of Urology, vol. 36, no. 1, p. 29–37, 2010 14. Pruett T, Simmons R. Status of percutaneous catheter drainage of abscesses. Surg Clin North Am 1988;68:89- 105. 14. Pruett T, Simmons R. Status of percutaneous catheter drainage of abscesses. Surg Clin North Am 1988;68:89- 105. 15. L. Pasqualini, C. Leli, GVL De Socio et al., “Retroperitoneal abscess : a rare localization of tuberculosis infection”, Infezionien medicine, vol. 16, no. 4, p. 230–232, 2008. p 16. RJ Barbalinardo, GB Hamilton, GR Eliot, EJ Lazaro, and C. Haycock, "Tuberculous retroperitoneal lymphadenopathy mimicking metastatic pancreatic carcinoma," Journal of the National Medical Association, vol. 78, no. 5, pp. 385–387, 1986 . pp 17. Lazare A, Thilagar B. Abdominal tuberculosis. Say Mon. 2007; 53:32-38. 28	34,192
https://openalex.org/W2975867678	OpenAlex	Open Science	CC-By	2,019	The role of idealisations in describing an isolated molecule	Vanessa A. Seifert	English	Spoken	10,127	15,111	Foundations of Chemistry (2020) 22:15–29 https://doi.org/10.1007/s10698-019-09342-7 Foundations of Chemistry (2020) 22:15–29 https://doi.org/10.1007/s10698-019-09342-7 Abstract The investigation of the relation between chemistry and quantum mechanics includes examining how the two theories each describe an isolated molecule. This paper focuses on one particular characteristic of chemistry’s and quantum mechanics’ descriptions of an isolated molecule; namely on the assumptions made by each description that an isolated molecule is stable and has structure. The paper argues that these assumptions are an ide- alisation. First, this is because stability and structure are partially determined by factors that concern the context in which a molecule is considered (i.e. thermodynamic condi- tions, time-range of experiment, environment, etc.). Secondly, the stability and structure of a molecule can only be empirically identified with reference to those factors. This paper examines these assumptions in the context of the philosophical literature on idealisations. This examination is a novel contribution that raises interesting questions about the relation between the two theories, the nature of stability and structure, and the function of these assumptions in the two theories. Keywords Idealisations · Relation between chemistry and quantum mechanics · Stability · Molecular structure * Vanessa A. Seifert vs14902@bristol.ac.uk The role of idealisations in describing an isolated molecule Vanessa A. Seifert1 Published online: 24 September 2019 © The Author(s) 2019 Published online: 24 September 2019 © The Author(s) 2019 1 University of Bristol, Bristol, UK The philosophical investigation of idealisations Making idealisations is one of the most common practices in science and numerous cases can be invoked as examples. ‘(F)rictionless planes, point masses, infinite velocities, iso- lated systems, omniscient agents, and markets in perfect equilibrium’ are regarded as idealisations that are made in the relevant theories or models (Frigg and Hartmann 2012, Section 1.1). Assuming that gas molecules are infinitely small, or that light consists of ‘one-dimensional beams rather than waves’ are also examples of idealisations (Strevens 2017, p. 2). The philosophy of science examines various questions regarding idealisations. For example, it examines the nature of idealisations and whether there are distinct kinds of idealisations in science. Moreover, it examines how particular figures in the history of sci- ence have understood idealisations. This is because idealisations are crucial to the work of central figures such as Aristotle, Galileo, and Newton (McMullin 1985, pp. 240–262). For example, McMullin analyses Galileo’s understanding of idealisations in order to identify a particular type of idealisation; namely Galilean idealisation (1985). Furthermore, the use of idealisations in science is often examined in relation to philosophical topics such as sci- entific realism, emergence, models in science, representation and explanation (Ladyman 2008, pp. 360–365). For example, the use of idealisations in science is used as putative evidence for antirealism (Weisberg 2007, p. 657; Cartwright 1983). On the other hand, it is also argued that idealisations can support scientific realism (Weisberg 2007, p. 657; Norton 2012, p. 211). Perhaps as is to be expected, there is no standardly accepted single account of idealisa- tions. As Weisberg states: Philosophers of science increasingly recognize the importance of idealization (…). Yet this recognition has not yielded consensus about the nature of idealization. The literature of the past 30 years contains disparate characterizations and justifications, but little evidence of convergence towards a common position. (2007, p. 639) The lack of a common position about idealisations becomes apparent from the fact that numerous definitions of idealisations are given. For example, Frigg and Hartmann propose the following definition: An idealization is a deliberate simplification of something complicated with the objective of making it more tractable. (2012, Section 1.1) McMullin defines idealisations in a similar manner; namely as ‘a deliberate simplifying of something complicated (a situation, a concept, etc.) with a view to achieving at least at a partial understanding of that thing’ (1985, p. 248). Introduction When chemistry and quantum mechanics each describe an isolated molecule, they assume that the molecule is stable and has structure. Identifying these assumptions as an idealisa- tion is a novel contribution to the discussion of idealisations in chemistry and in quantum mechanics, and raises interesting philosophical issues, including those concerning the rela- tion between chemistry and quantum mechanics. The paper is structured as follows. “The philosophical investigation of idealisations” section presents how idealisations are understood and discussed in philosophy. “Stability and structure in chemistry and quantum mechanics” section explains why regarding an iso- lated molecule as being stable and having structure is an idealisation. When it comes to describing an isolated molecule, both chemistry and quantum mechanics make this ide- alisation. “Philosophical implications” section presents how this idealisation contributes 56789) 3 23451 V. A. Seifert 16 to the investigation of various philosophical issues. Lastly, “Conclusion” section concludes this paper by restating the paper’s main claim. to the investigation of various philosophical issues. Lastly, “Conclusion” section concludes this paper by restating the paper’s main claim. The philosophical investigation of idealisations Norton argues that idealisations should be taken to ‘refer to new systems some of whose properties approximate some of those of the target system’ (Norton 2012, p. 207), whereas Weisberg defines idealisations as ‘the intentional introduction of distortion into scientific theories’ (Weisberg 2007, p. 639). These definitions illustrate that idealisations are ‘a feature of both the formulation of laws and theories and of their application to the world’ (Ladyman 2008, p. 358). For example, Weisberg understands idealisations as a feature of scientific theories, whereas 1 3 The role of idealisations in describing an isolated molecule 17 Norton understands them as a feature of the system or phenomenon that scientific theories describe.1 Norton understands them as a feature of the system or phenomenon that scientific theories describe.1 A question which is raised about idealisations concerns their function in science. According to Frigg and Hartmann, the function of idealisations is to make ‘something’ more ‘tractable’ (2012, Section 1.1). Other philosophers specify in more detail the function of idealisations. For example, idealisations are used in order to: (1) ‘reduce the complexity of the model’, by either ‘making the mathematics more tractable or by reducing the empirical demands of the model’ (Strevens 2017, p. 3); (1) ‘reduce the complexity of the model’, by either ‘making the mathematics more tractab ‘reduce the complexity of the model’, by either ‘making the mathematics more tractable (1) ‘reduce the complexity of the model’, by either ‘making the mathematics more tractable or by reducing the empirical demands of the model’ (Strevens 2017, p. 3); p y y g or by reducing the empirical demands of the model’ (Strevens 2017, p. 3); (2) identify the ‘core or primary causal factors that give rise to the phenomenon of interest’ (Weisberg 2007, p. 651); and, (3) construct ‘a single model for a particular target or class of target phenomena’ (Weisberg 2007, p. 655). Moreover, a particular notion of idealisation is often supported with respect to the descrip- tion of a particular phenomenon. For example, Batterman proposes the notion of ‘infinite idealizations’ with respect to phase transitions and critical phenomena (2011, p. 1033). Weisberg proposes the notion of ‘multiple-models idealization’ regarding the description of global weather patterns (2007, p. 646). McMullin argues that the application of geom- etry for the specification of the movement of planets is an example of ‘mathematical ideali- zation’ (1985, pp. 248–254). 3 Specifically, ‘some scale models would also qualify as idealized models and it is not clear where exactly to draw the line between idealized and analogue models’ (Frigg and Hartmann 2012, Section 1.1). This question requires a detailed analysis of models and representations in science and is not examined here (for example Ladyman 2008, pp. 360–364). 1 McMullin, as well as Frigg and Hartmann, formulate a definition which allows both understandings idealisations; namely as a feature of theories and as a feature applied to systems or phenomena in the wor 1 McMullin, as well as Frigg and Hartmann, formulate a definition which allows both understandings of idealisations; namely as a feature of theories and as a feature applied to systems or phenomena in the world. 2 The former is supported by Norton (2012). An example of the latter is (Wayne 2012, p. 343). A similar issue arises with respect to the distinction between idealisations and abstractions (Cartwright 1989; Lady- man 2008, pp. 362–363). 3 Specifically, ‘some scale models would also qualify as idealized models and it is not clear where exactly to draw the line between idealized and analogue models’ (Frigg and Hartmann 2012, Section 1.1). This question requires a detailed analysis of models and representations in science and is not examined here (for example Ladyman 2008, pp. 360–364). 1 McMullin, as well as Frigg and Hartmann, formulate a definition which allows both understandings of idealisations; namely as a feature of theories and as a feature applied to systems or phenomena in the world. 2 The former is supported by Norton (2012). An example of the latter is (Wayne 2012, p. 343). A similar issue arises with respect to the distinction between idealisations and abstractions (Cartwright 1989; Lady- man 2008, pp. 362–363). 3 S ifi ll ‘ l d l ld l lif id li d d l d i i l h l 1 McMullin, as well as Frigg and Hartmann, formulate a definition which allows both understandings of idealisations; namely as a feature of theories and as a feature applied to systems or phenomena in the world. 2 The former is supported by Norton (2012). An example of the latter is (Wayne 2012, p. 343). A similar 2 The former is supported by Norton (2012). An example of the latter is (Wayne 2012, p. 343). A sim issue arises with respect to the distinction between idealisations and abstractions (Cartwright 1989; La man 2008, pp. 362–363).i The philosophical investigation of idealisations There are disagreements about several issues regarding idealisations. For example, some argue that idealisations are distinct from approximations, whereas others use the two terms interchangeably.2 Also, there are particular examples of theories, models, or repre- sentations for which it is not very clear whether idealisations or some other notion (such as analogues) accurately specifies their ‘representational styles’ (Frigg and Hartmann 2012, Section 1.1).3 Another issue that is discussed concerns the categorisation of idealisations. For example, Frigg and Hartmann argue that there are two main kinds of idealisations; i.e. ‘Galilean’ and ‘Aristotelian idealisations’ (2012, Section 1.1). McMullin distinguishes between ‘mathematical’, ‘Galilean’ (which are further distinguished into ‘construct’ and ‘causal idealisations’), ‘material’, ‘formal’ and ‘subjunctive idealisations’ (McMullin 1985; Ladyman 2008, pp. 361–362). Weisberg argues that there are three kinds of idealisations; namely ‘Galilean’, ‘minimalist’, and ‘multiple-models idealizations’ (2007). Strevens examines ‘asymptotic’ and ‘simple idealisations’ (2017). Whether there is a unique clas- sification that underwrites all proposed kinds of idealisations is not examined here. Regard- less of the exact characterisation of ‘idealisation’ and its various species and cognates, the 1 3 3 3 V. A. Seifert 18 way chemistry and quantum mechanics describe the stability and structure of an isolated molecule counts as idealisation of some kind in the sense of all the accounts above.4 way chemistry and quantum mechanics describe the stability and structure of an isolated molecule counts as idealisation of some kind in the sense of all the accounts above.4 Lastly, a number of examples from chemistry and quantum mechanics have been invoked when discussing idealisations in science. For example: 1. The use of approximate wavefunctions in order to calculate the properties of molecules, is regarded as an example of Galilean idealisation (Weisberg 2007, p. 641). 2. Treating ‘the vibrating bond as spring-like with a natural vibrational frequency’ in order to calculate the vibrational properties of a covalent bond, is regarded as a minimalist idealisation (Weisberg 2007, p. 644). 3. The description of atoms and molecules via the molecular orbital or the valence bond approach is regarded as an example of Weisberg’s third kind of idealisation; i.e. the multiple-model idealisation (Weisberg 2007, p. 646). 4. The Bohr image of the atom is regarded as an example of Galilean idealisation (McMul- lin 1985, p. 260). 5. The Lewis electron pair model of chemical bonding is also regarded an idealisation (Weisberg 2007, p. 650). 5 This paper leaves it open whether there are other features in chemistry or quantum mechanics which have not been considered as idealisations in the philosophical literature and which could possibly be construed as such. 6 A central reference of this paper is the Gold Book (IUPAC 2014) which is provided by the International Union of Pure and Applied Chemistry (IUPAC). IUPAC is one of the leading authorities regarding chem- ical nomenclature and the definition of chemical terms. The glossary of the Gold Book is more or less accepted by the entire chemical community; the terms and their respective definitions have been rigorously reviewed and published in chemistry journals. 4 The particular example may be alternatively identified as an abstraction or approximation. This is not examined here. The philosophical investigation of idealisations While this paper does not examine the general role of idealisations in chemistry and in quantum mechanics, the above examples illustrate that both chemistry and quantum mechanics employ various idealisations of possibly different types. Moreover, some of the above examples illustrate that there are particular features of the quantum mechani- cal description of an isolated molecule which have been examined in terms of the notion of idealisations. Nevertheless, there is a particular feature of quantum mechanics (and of chemistry) that has not been identified as an idealisation in either the philosophy of science or the philosophy of chemistry literature; namely the assumptions that an isolated molecule is stable and has structure.5 Stability and structure in chemistry and quantum mechanics This section presents how stability and structure are defined, understood, and empirically identified.6 This analysis explicates why the assumption of an isolated molecule with sta- bility and structure is an idealisation. Specifically, this assumption is correctly construed as an idealisation for two main reasons. First, stability and structure are partially determined by factors that concern the context in which a molecule is considered (i.e. thermodynamic 1 3 The role of idealisations in describing an isolated molecule 19 conditions, time-range of experiment, environment, etc.).7 Secondly, the stability and struc- ture of a molecule can only be empirically identified with reference to those factors. conditions, time-range of experiment, environment, etc.).7 Secondly, the stability and struc- ture of a molecule can only be empirically identified with reference to those factors. i Note that a particular account of idealisations regarding their nature, function, etc. is not currently assumed. The aim of this section is to present the scientific evidence which sup- ports the claim that this assumption is an idealisation. 7 Llored (2012) also points out factors that are involved in the empirical identification and description of molecules. However, contrary to the present paper, those factors are taken by Llored to include the relevant tools, methods, instruments and modes of access. Given this, as well as that he does not discuss this issue with respect to idealisations, this paper does not further consider his account. 10 The term ‘conformation’ standardly applies to molecules which exhibit different stereoisomeric struc- tures. However, it is also understood more broadly as referring to the different possible spatial arrangements of the atoms that comprise a molecular entity. The latter understanding of conformation is assumed here. 7 Llored (2012) also points out factors that are involved in the empirical identification and description of molecules. However, contrary to the present paper, those factors are taken by Llored to include the relevant tools, methods, instruments and modes of access. Given this, as well as that he does not discuss this issue with respect to idealisations, this paper does not further consider his account. 8 What is meant by ‘isolation’ is explained later on in this paper. 9 A chemical species is ‘(a)n ensemble of chemically identical molecular entities that can explore the same set of molecular energy levels on the time scale of the experiment’ (IUPAC 2014, p. 264). 10 The term ‘conformation’ standardly applies to molecules which exhibit different stereoisomeric struc- tures. However, it is also understood more broadly as referring to the different possible spatial arrangements of the atoms that comprise a molecular entity. The latter understanding of conformation is assumed here. 9 A chemical species is ‘(a)n ensemble of chemically identical molecular entities that can explore the same set of molecular energy levels on the time scale of the experiment’ (IUPAC 2014, p. 264). 10f Stability There are two cases in which stability is considered with respect to a single molecule, namely: a. the stability of a molecule when considered as part of an ensemble; and, b. the stability of a molecule in isolation. IUPAC (2014) does not provide a definition of stability with reference to a molecule (whether isolated or not).8 However, it is common practice in chemistry to talk about a molecule’s stability. Regarding the stability of a molecule that is part of an ensemble, a molecule is stable in virtue of being part of a stable chemical species, and not in virtue of being in its ground state.9 This is because the molecule that is part of an ensemble is neither static, nor always fixed at its ground state. A molecule that is part of an ensem- ble interacts both intermolecularly and intramolecularly, and its conformation dynamically changes over time.10 Therefore, when one refers to a molecule (which is part of an ensem- ble) as being stable, what is meant is that the molecule is part of a stable chemical species. Given this, it follows that the stability of a molecule is partially determined by the fac- tors that determine the stability of the chemical species of which the molecule is part. These factors become apparent from the definition of stability with reference to chemical species: As applied to chemical species, the term expresses a thermodynamic property, which is quantitatively measured by relative molar standard Gibbs energies. A chemical species A is more stable than its isomer B if ΔrGo > 0 for the (real or hypothetical) reaction A ≥ B, under standard conditions. If for the two reactions: ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. P ≥X + Y(ΔrGo 1 ) Q ≥X + Z(ΔrGo 2 ) P ≥X + Y(ΔrGo 1 ) Q ≥X + Z(ΔrGo 2 ) 8 What is meant by ‘isolation’ is explained later on in this paper. 9 9 A chemical species is (a)n ensemble of chemically identical molecular entities that can explore the same set of molecular energy levels on the time scale of the experiment’ (IUPAC 2014, p. 264). 10 The term ‘conformation’ standardly applies to molecules which exhibit different stereoisomeric struc- tures. However, it is also understood more broadly as referring to the different possible spatial arrangements of the atoms that comprise a molecular entity. The latter understanding of conformation is assumed here. ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. ( r 2 ) ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. 7 Llored (2012) also points out factors that are involved in the empirical identification and description of molecules. However, contrary to the present paper, those factors are taken by Llored to include the relevant tools, methods, instruments and modes of access. Given this, as well as that he does not discuss this issue with respect to idealisations, this paper does not further consider his account. 8 10 The term ‘conformation’ standardly applies to molecules which exhibit different stereoisomeric struc- tures. However, it is also understood more broadly as referring to the different possible spatial arrangements of the atoms that comprise a molecular entity. The latter understanding of conformation is assumed here. 1 3 3 V. A. Seifert 20 Both in qualitative and quantitative usage the term stable is therefore always used in reference to some explicitly stated or implicitly assumed standard. (IUPAC 2014, p. 1432) Both in qualitative and quantitative usage the term stable is therefore always used in reference to some explicitly stated or implicitly assumed standard. (IUPAC 2014, p. 1432) Based on the above, the stability of a molecule which is part of a chemical species is rela- tive to the following factors: Based on the above, the stability of a molecule which is part of a chemical species is rela- tive to the following factors: • The particular reaction(s) with respect to which the stability of the relevant chemical species is considered. For example, while a chemical species P is more stable relative to Y when considered with respect to how stable Q is relative to Z, P is less stable rela- tive to Y when considered with respect to how stable A is relative to T (for the hypo- thetical reaction A ≥ X + T, ΔrGo 3 , where ΔrGo 3 > ΔrGo 1). 3 3 1 • The particular chemical species with which the relevant chemical species’ stability is relative. 13 Intrinsic properties are ‘properties objects have just in virtue of how they are in themselves, not how they are in relation to other things’ (Ney 2014, p. 285). 11 For example (Marvel 2009) examines stability with respect to various thermodynamic factors, including temperature. 12 Ney’s definition of relational properties provides a rather general understanding of relational properties because it does not specify what renders an object in relation to something else. This understanding of relational properties suffices in order to support the claim that the stability of a molecule that is part of an ensemble is not a property that the molecule has in and of itself. 11 For example (Marvel 2009) examines stability with respect to various thermodynamic factors, including temperature.i 14 To my knowledge, there is no bibliographical reference in chemistry or quantum chemistry which explic- itly specifies what renders a molecule in isolation (for example IUPAC 2014). However, based on how chemists and quantum chemists describe an isolated molecule and on how physics defines isolation, I take this to be a correct definition of an isolated molecule. 15 One could justify this assumption if, for example, the chemical substance is examined in very low pres- sure. 16 In fact, there has been extensive development of techniques and experiments in chemistry which aim at measuring the value of properties of single molecules, instead of inferring the properties of a single mol- ecule by measuring the properties of an ensemble of molecules. This research is often referred to as ‘single- molecule chemistry’ (Bai et al. 1999; Walter 2008). ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. ( r 2 ) This is because the ground state of a molecule is taken to correspond to a particular internal structure of 1 3 The role of idealisations in describing an isolated molecule 21 the molecule. This structure corresponds to the average nucleonic and electronic configu- rations of the atomic nuclei and electrons within the molecule, and is determined by their respective interactions. However, the in principle impossibility of empirically identifying any property of an isolated system, entails that this model of the stability of an isolated molecule (specified solely in terms of the molecule’s ground state) is highly idealised.ii the molecule. This structure corresponds to the average nucleonic and electronic configu- rations of the atomic nuclei and electrons within the molecule, and is determined by their respective interactions. However, the in principle impossibility of empirically identifying any property of an isolated system, entails that this model of the stability of an isolated molecule (specified solely in terms of the molecule’s ground state) is highly idealised.ii i In order to support this claim, it is important to first define what is meant by ‘isolation’. The term is understood in science in various ways; there is thermodynamic isolation, mate- rial isolation, radiative isolation, etc. Given how science understands ‘isolation’, a mol- ecule is understood here as a system in isolation when the following requirements are met: • The molecule is far removed from any other system and thus doesn’t interact with other molecular entities (i.e. with other atoms, molecules, ions, etc.); and, 14 • The molecule is far removed from any other system and thus doesn’t interact with other molecular entities (i.e. with other atoms, molecules, ions, etc.); and, • The total energy of the molecule is conserved.14 This is a highly idealised representation of a molecule for three reasons. First, it is very difficult to empirically examine a molecule’s properties in an environment in which the molecule doesn’t interact with other molecular entities. In practice, when it comes to the empirical support of chemical properties, the experiments are performed on chemical sub- stances and not on a single molecular entity. Given this, it is common in chemical practice to assign chemical properties to a single molecule by measuring properties of samples of matter. ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. ( r 2 ) For example, while P is more stable relative to Y (when considered with the reaction Q ≥ X + Z), it is less stable relative to M (when considered with the reaction Q ≥ X + Z), if it is the case that there is a hypothetical reaction P ≥ X + M (with ΔrGo 4 ) such that ΔrGo 4 < ΔrGo 2. 4 2 • The thermodynamic conditions in which those reactions are performed. The relative molar Gibbs energies are calculated and compared under the assumption that the con- sidered reactions are performed under standard thermodynamic conditions. However, if the thermodynamic conditions are different (for example, the temperature or pressure of the examined systems are different), then it is possible that the relevant molar Gibbs energies are different, and thus that the stability of the chemical species is different.11 ff • The environment in which that particular set of reaction(s) is assumed to be performed. This includes the specification of the type of solution or background gas within which the reactions are performed. For example, there are polymers which ‘are reported to be stable in air up to 500 °C and to nearly 800 °C in nitrogen’ (Marvel 2009, p. 361). Given the above, the stability of a molecule that is part of an ensemble is a relational property of that molecule; namely a property which the molecule has ‘in relation to other things’ (Ney 2014, p. 285).12 This is because the molecule has this property in virtue of the relation that the relevant chemical species has with other chemical species, as well as in relation to particular thermodynamic conditions, to a set of reaction(s), and to the environ- ment in which the relevant chemical species is assumed to be found. The other case considered here is a molecule in isolation. Contrary to a molecule which is part of an ensemble, one would expect the stability of an isolated molecule to be an intrinsic property which is only determined by its composing entities and interactions.13 In this case, the stability of a molecule in isolation would refer to the ground state of the mol- ecule; namely to ‘the state of lowest Gibbs energy’ (IUPAC 2014, p. 646). ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. ( r 2 ) The experiments are performed in such conditions that it is safe to assume that the behaviour of each molecule of an ensemble, closely resembles the behaviour of a single molecule when in isolation.15fi This is merely an epistemic difficulty which, with the improvement of technological means, could possibly be overcome.16 There is, however, a second reason why the descrip- tion of an isolated molecule is highly idealised. The empirical identification of a mole- cule’s stability (or of any other property) necessitates the interaction of that molecule with an experimental device, through the emission of light or the imposition of a field to the sys- tem. Therefore, the properties of an examined system are necessarily those that the system exhibits when it is in a state of non-conservation of energy. This cannot be overcome; in principle any act of measurement will probe the examined system in such a way that it will cease to be in isolation. Thirdly, there is one additional feature that renders the chemical and quantum mechan- ical descriptions of an isolated molecule an idealisation; namely the way in which they represent it in time. To the extent that the stability of an isolated molecule is taken to correspond to its ground state, the stability of an isolated molecule is defined independ- ent of time. This is because the ground state of a molecule is a stationary state; namely a state that ‘does not evolve with time’ (IUPAC 2014, p. 1443). Taking stability to be a 1 3 3 V. A. Seifert 22 time-independent property is an idealisation because in practice the stability of a molecule is always relevant to the particular time range of an experiment. In fact, there is empiri- cal evidence of particular molecules which change from one structure into another in the course of time, suggesting that a molecule may not remain in one stable state indefinitely. For example (Meléndez-Martínez et al. 2014) examine the time-dependence of the differ- ent isomeric structures of carotenoids. So, given that in non-isolation, stability is a time- dependent property, it may be assumed that stability in isolation is a time-dependent prop- erty as well. On the other hand, one can assume that the stability of an isolated molecule is time-independent, in accordance to how it is described in quantum mechanics through the stationary states. While both assumptions are consistent with empirical evidence, the above analysis suggests the following. ΔrGo 1 > ΔrGo 2 , P is more stable relative to the product Y than is Q relative to Z. ( r 2 ) When describing an isolated molecule, chemistry’s and quantum mechanics’ understanding of stability as a time-independent property is an ideali- sation; namely something that is assumed, rather than empirically identified.i i In sum, the stability of a molecule is empirically verified always in relation to particular factors, including the thermodynamics conditions, the environment, and the time-range of an experiment. Given this, the stability of an isolated molecule refers to a state of the mole- cule which can never be empirically identified and whose existence is assumed, rather than empirically verified. This idealisation is made both in chemistry and in quantum mechan- ics, whenever the two theories describe a stable isolated molecule. The next section argues that a similar idealised understanding is assumed with respect to the structure of an iso- lated molecule. Structure Molecular structure is a central property of molecules that figures in the explanation of their chemical behaviour. The particular structure of a molecule can explain (to a certain degree) the stability and reactivity of that molecule. Also, the structure of a molecule that participates in a reaction may partially explain the process through which it transforms into particular products (i.e. the reaction mechanism), as well as the sort of products that are produced during the reaction. Moreover, physical properties of matter may be explained in terms of the structure of the molecules that compose it.i While molecular structure is employed by IUPAC for the definition of other chemical terms (such as molecular modelling and the Lewis formula), the term itself is not defined in the Gold Book. In light of this, this paper proposes a definition which is based on how the term is broadly understood in chemistry. Moreover, similarly to the examination of stability, this paper investigates how a molecule’s structure is understood and empirically identified for two cases, namely: The role of idealisations in describing an isolated molecule The role of idealisations in describing an isolated molecule 23 Given the above, the structure of a molecule is defined here as the spatial arrange- ment of the atoms that constitute it. The factors that determine this spatial arrangement are contingent not only on the identity and interactions between the atoms that comprise the molecule, but also on the context in which the molecule as a whole is considered. This becomes evident when examining the structure of a molecule that is considered as part of an ensemble. In this case, the spatial arrangement of a molecule’s atoms is determined by (i) the interactions between the molecule’s own atoms (i.e. the intramolecular interactions), and (ii) the interactions between the molecule’s atoms and one or more distinct molecular entities (i.e. the intermolecular interactions).17 Molecular structure is a collective term in the sense that it doesn’t refer to one par- ticular and empirically measurable chemical property, but rather to a collection of empiri- cally measurable chemical (but also quantum mechanical) properties of the molecule. This seems to be in accordance with intuition. For example, when describing the structure of a box, one specifies its angles, the length of its sides, the distance between its surfaces, etc. There is no one measurable property that fully identifies and describes the structure of a box. The same applies to a molecule’s structure. There is a set of properties that need to be specified in order to provide a satisfactory description of molecular structure. i There is no one measurable property that fully identifies and describes the structure of a box. The same applies to a molecule’s structure. There is a set of properties that need to be specified in order to provide a satisfactory description of molecular structure. i Given this, the properties that specify a molecule’s structure when that molecule is part of an ensemble, can be categorised into two sets. The first set includes the information that specifies the intramolecular interactions. This includes reference to: • the types of intramolecular interactions, both bonding and nonbonding; • the types of intramolecular interactions, both bonding and nonbonding; the properties that are assigned to the intramolecular interactions; certain properties that are based on the quantum mechanical description of molecules; the pictorial representation of molecules; and,f p p the properties that specify the structural differences between molecules. 17 This justifies why this paper distinguishes between the structure of a single molecule considered as part of an ensemble, and the structure of a single molecule in isolation. 18 Needham raises a similar point regarding the properties of a molecule: ‘A molecule in the liquid isn’t like a water molecule in isolation (in the gas phase at very low pressure), but will be subject to all the stresses imposed by the ambient medium’ (2017, p. 135). 17 This justifies why this paper distinguishes between the structure of a single molecule considered as part of an ensemble, and the structure of a single molecule in isolation. 18 Needham raises a similar point regarding the properties of a molecule: ‘A molecule in the liquid isn’t like a water molecule in isolation (in the gas phase at very low pressure) but will be subject to all the stresses 17 This justifies why this paper distinguishes between the structure of a single molecule considered as part of an ensemble, and the structure of a single molecule in isolation. 18 o a e se b e, a d t e st uctu e o a s g e o ecu e so at o . 18 Needham raises a similar point regarding the properties of a molecule: ‘A molecule in the liquid isn’t like a water molecule in isolation (in the gas phase at very low pressure), but will be subject to all the stresses imposed by the ambient medium’ (2017, p. 135). a. a single molecule which is part of an ensemble; and, b. a single molecule in isolation. a. a single molecule which is part of an ensemble; and, b. a single molecule in isolation. a. a single molecule which is part of an ensemble; and, b. a single molecule in isolation. Two concepts are often employed when referring to molecular structure; ‘shape’ and ‘conformation’. The ‘structure of a molecule’ is often employed interchangeably with the ‘shape of a molecule’ or ‘molecular shape’ (Ramsey 1997; Woolley 1976, p. 28). Moreo- ver, depending on the type of molecules that are examined, molecular structure is often referred to in terms of the conformations that a specific molecule can take. Furthermore, the specific structure of a particular molecule is described by specifying the spatial arrangement of the atoms within that molecule. 1 3 • the properties that are assigned to those interactions. This set is dependent on the identity of the molecule under examination, but also on the identity of the surrounding molecular entities. Depending on the explanatory, predictive, and heuristic needs of chemistry as well as on the particular type of molecule that is under examination, intermolecular interactions may be disregarded in the description of a molecule’s structure. For example, a molecule with aromatic character exhibits unusual stability due to its structure, regardless of the particular environment in which it is considered (IUPAC 2014, p. 109). Therefore, its structure and stability are usually explained independently of the environment and conditions in which the molecule may be found. On the other hand, there are also cases where intermolecu- lar interactions play an important role in explaining molecular structure, and thus in the behaviour of molecules. For example, the ‘abnormal physical properties’ of matter com- posed of NH3, H2O or HF is explained with reference to the intermolecular interactions that take place between such molecules (namely in terms of hydrogen bonding) (Needham 2013, p. 52). In light of the above, it is clear that when a molecule is considered as part of an ensem- ble then its structure is a relational property, in the sense that a molecule’s structure is par- tially determined by the relations it develops with other molecular entities. y y p Concerning an isolated molecule, it becomes evident that, in virtue of being isolated, intermolecular interactions do not play any role in the determination of the molecule’s structure. However, it is not entirely accurate to infer from this that what determines an isolated molecule’s structure are only its intramolecular interactions. Just like in the case of stability, molecular structure is empirically supported in the context of chemical spe- cies, and not of an isolated molecule. The terms employed for the description of molecu- lar structure are defined with reference to chemical species and empirically supported by experiments done on ensembles of molecular entities. 19 For example, depending the temperature of the system, the molecule may exhibit different structures (Xiao et al. 2016). 20 A molecule has the property of being chiral when its mirror molecule is not identical to itself. The term ‘enantiomer’ refers to each molecule of any pair of chiral molecules. The two enantiomers of a pair of chiral molecules are often referred to as the right- and left-handed enantiomers. The role of idealisations in describing an isolated molecule Moreover, the structure of a molecule can be significantly affected by its intermolecular interactions. For example, the helical structure of DNA is determined by the intermolecu- lar interactions (mainly hydrogen bonds) between the nucleic acids of the two strands that make up the DNA. These two strands are distinct molecules, and the reason why these two strands curl up into the overall helical structure of DNA (and why therefore they acquire their particular structure) is due to the intermolecular interactions between them. Another example is the structure of a water molecule (H2O). Two water molecules in a water dimer [i.e. (H2O)2] do not have the same structure and each molecule’s structure in the water dimer is also different from the structure of a single water molecule (whether in gas-phase, liquid-phase or solid-phase water) (Klopper et al. 2000).18 In light of this, there is a second set of properties which are invoked for the specification of a molecule’s structure. This set includes reference to: • the types of interactions between the molecule (or its constituting atoms) and one or more distinct molecular entities; and, • the types of interactions between the molecule (or its constituting atoms) and one or more distinct molecular entities; and, 1 3 3 V. A. Seifert 24 • the properties that are assigned to those interactions. 20 A molecule has the property of being chiral when its mirror molecule is not identical to itself. The te ‘enantiomer’ refers to each molecule of any pair of chiral molecules. The two enantiomers of a pair of ch molecules are often referred to as the right- and left-handed enantiomers. 19 For example, depending the temperature of the system, the molecule may exhibit different structu (Xiao et al. 2016). • the properties that are assigned to those interactions. Moreover, the structure of a single molecule is partially determined by the thermodynamic conditions in which the molecule is considered, as well as by the environment in which it is found.19 Therefore, the structure of an isolated molecule refers to an idealised state of the molecule which can never be empirically identified and whose existence is assumed rather than empirically verified.i ii Consider, for example, the empirical identification of chirality; namely of the ‘geometric property’ of a molecule ‘of being non-superposable on its mirror image’ (IUPAC 2014, p. 269).20 A chiral molecule is experimentally identified through the measurement of the opti- cal activity of the sample in which that molecule is contained (IUPAC 2014, p. 1030). Any assembly of chiral molecules that is either enantiomerically pure (i.e. it contains only right or only left-handed enantiomers) or that contains an unequal amount of a pair of enantiom- ers (i.e. it contains, say, more left- than right- handed enantiomers) is optically active. The measurement of optical activity is performed on a sample of material, i.e. on matter com- posed of an ensemble of (either chemically distinct or identical) molecular entities. While chemists infer from such experiments that the observed sample contains molecules with 1 3 The role of idealisations in describing an isolated molecule 25 specific chiral structure, the observed optical activity is that of the entire sample, and not of a single chiral molecule. specific chiral structure, the observed optical activity is that of the entire sample, and not of a single chiral molecule. It should be noted that there are advances on single-molecule chemistry concerning the measurement of the polarisation of light produced by one single molecule. This is directly relevant to the empirical examination of the chirality of a single molecule (Chuntonov and Haran 2013). However, even in single-molecule chemistry, the single molecule whose properties are measured, is considered in a system where it interacts with other molecules. It is not the case that the single molecule is far removed from other molecular entities. Therefore, both the presence of other molecules, as well as the environmental and thermo- dynamic conditions in which a molecule is considered, are still relevant to the determina- tion of its structure. An additional idealisation concerning the structure of a molecule becomes apparent from how structure is pictorially represented. • the properties that are assigned to those interactions. In chemistry, the structure of a molecule is represented statically, as if the molecule and its constituting atoms are not continuously moving and rotating in space. However, the pictorial representation of a molecule’s struc- ture only highlights the average shape of a molecule, and does not refer to a static and unchanging structure. This is important because, even if one considers an isolated mol- ecule, it is still the case that the particles that comprise an isolated molecule interact with each other and move in space. This is due to the existence of intramolecular interactions which persist even when the total energy of the system is constant (since the angular momentum of its comprising entities is non-zero). Therefore, the structure of a molecule is dynamic in the sense that a molecule (even in isolation) can possess different conforma- tions in time as a result of the continuous interactions of the entities (namely electrons and nuclei) that comprise the molecule (Longuet-Higgins 1963, p. 446). There is one additional factor that plays a central role in the understanding of struc- ture; namely time. The empirical identification of a molecule’s structure is only possible if the examined molecule is stable for a particular range of time. If a molecule is not stable under particular conditions and within a particular environment, then it is not possible to empirically identify its structure. Given that the empirical identification of the stability of a molecule is relevant to the time-range of an experiment, it follows that the empirical identification of a molecule’s structure is also relevant to that particular time-range. This is further supported by the fact that there are types of molecules whose structure changes in the course of time (Meléndez-Martínez et al. 2014). All the above is not taken into account in quantum mechanics which describes the properties of a molecule by specifying its sta- tionary (i.e. time-independent) states. All the above illustrate the idealised framework in which chemistry and quantum mechanics each describe the structure of an isolated molecule. Philosophical implications For example, one could say that this is an example of Gali- lean idealisation as the latter is defined by Weisberg: Galilean idealization is the practice of introducing distortions into theories with the goal of simplifying theories in order to make them computationally tractable. One starts with some idea of what a nonidealized theory would look like. Then one men- tally and mathematically creates a simplified model of the target. (2007, p. 640) Galilean idealization is the practice of introducing distortions into theories with the goal of simplifying theories in order to make them computationally tractable. One starts with some idea of what a nonidealized theory would look like. Then one men- tally and mathematically creates a simplified model of the target. (2007, p. 640) Alternatively, one could say that this is an example of Aristotelian idealisation, as defined by Frigg and Hartmann: Aristotelian idealization amounts to ‘stripping away’, in our imagination, all proper- Aristotelian idealization amounts to ‘stripping away’, in our imagination, all proper- ties from a concrete object that we believe are not relevant to the problem at hand. This allows us to focus on a limited set of properties in isolation. (2012, Section 1.1) Aristotelian idealization amounts to ‘stripping away’, in our imagination, all proper- ties from a concrete object that we believe are not relevant to the problem at hand. This allows us to focus on a limited set of properties in isolation (2012 Section 1 1) This allows us to focus on a limited set of properties in isolation. (2012, Section 1.1) While it is possible to argue for different types of idealisations, there are particular types that do not apply to the idealisation presented here. For example, the idealisation is not a case of an infinite idealisation where the latter is defined as ‘distortions or misrepresenta- tions (…) in which some parameter takes on an infinite or infinitesimal value’ (Shech 2018, p. 1). In fact, it is not a case of a mathematical idealisation, in the sense that some math- ematical value that is involved in the description of the system, is set to infinity or zero. Consider, for example, McMullin’s definition of mathematical idealisation: Mathematical idealisation is a matter of imposing a mathematical formalism on a physical situation, in the hope that the essentials of that situation will lend them- selves to mathematical representation. (1985, p. Philosophical implications The previous section presented the factors that determine a molecule’s stability and struc- ture. A molecule’s stability and structure are by necessity empirically identified in rela- tion to those factors. Hence, when one describes a molecule in isolation, it is just assumed that it is stable and has structure, because this description (whether chemical or quantum mechanical) disregards the factors that partially determine the stability and structure of an empirically identifiable molecule. Whether this assumption is justified (namely whether an isolated molecule is stable and has structure) is not examined here. The paper only points 3 3 V. A. Seifert 26 out that, given how science empirically identifies these two properties, an isolated mol- ecule with stability and structure is an idealisation. out that, given how science empirically identifies these two properties, an isolated mol- ecule with stability and structure is an idealisation. If one takes into account the philosophical literature on idealisations, then numerous questions can be raised about the assumption that an isolated molecule is stable and has structure. First, there are various ways in which one could understand this idealisation. For example, one could define this as a deliberate simplification/distortion of the description of a molecule which disregards some of the factors that determine the molecule’s proper- ties, in order to achieve one or more goals (i.e. tractability, solubility, simplicity, generality, identification of core causal factors, etc.) (in line with McMullin 1985; Weisberg 2007). On the other hand, one could say that chemistry and quantum mechanics each refer to a new system (i.e. an isolated and time-independent molecule) some of whose properties approxi- mate some of those of the target system (i.e. a non-isolated and time-dependent molecule) (in line with Norton 2012). Contrary to both views, it could be argued that there is no inter- esting difference between these two interpretations of the idealisation; both interpretations capture the same understanding of idealisation regardless of whether we formulate it in terms of descriptions or in terms of the system to be described. However, there are philoso- phers who would possibly disagree with this. For example, Norton argues that his proposed understanding of idealisation introduces ‘reference to a novel system’ and thus carries ‘a novel semantic import’ (2012, pp. 208–209). Another question is whether the assumption that is presented here is an example of a particular type of idealisation. Philosophical implications 254) That the particular assumption is not a case of an infinite or mathematical idealisation is supported as follows. Indeed, quantum mechanics imposes a mathematical formalism on molecules via the Schrödinger equation. The equation assumes that an isolated molecule is stable and has structure by setting the values of some parameters in the Schrödinger equation. However, the same does not apply to chemistry’s description of a molecule. This 1 3 The role of idealisations in describing an isolated molecule 27 is because chemistry is not primarily a mathematical description; its description of the rel- evant phenomena includes non-mathematical vocabulary as well. While it employs math- ematical formalisms (from thermodynamics, quantum mechanics, etc.), it also employs a formalism that is non-mathematical. For example, chemistry’s definition and understand- ing of stability involves reference to chemical reactions which in turn are described with reference to the elements of the periodic table. This issue also raises the question of why this idealisation is made in chemistry and in quantum mechanics. Since this idealisation is not only made in the mathematical formal- ism of quantum mechanics, it is inadequate to claim that its function is the mathematical tractability of the system. Perhaps its function is simplicity in general, in the sense that one includes in the descriptions as little as possible (Weisberg 2007, p. 650). Alternatively, its function may also be explanatory because the descriptions employ this idealisation in order to ‘include only the core causal factors which give rise to a phenomenon’ (Weisberg 2007, p. 642). While this section does not examine and specify in detail what the function of this idealisation is, it is evident that this idealisation serves more than one function in the relevant descriptions. First, this idealisation accommodates the mathematical tractability of the quantum mechanical description of a molecule. This is because, if quantum mechanics took into account time and the different conditions in which the molecule can be consid- ered, then the relevant mathematical equation would be very difficult, if not impossible, to solve. Moreover, this idealisation has an explanatory function in the following sense. It is much easier for chemists to explain and understand the properties of a molecule, if they consider it in isolation and independent of time. Philosophical implications By considering a molecule in isolation, chemists identify particular properties of the molecule and explain how these properties account for macroscopic properties of the matter in which the molecule is part of. These properties play a large role in understanding why the molecule behaves the way it does, even if these properties are not the only factors which determine its behaviour (see also Weisberg 2007, p. 650). One more issue that is revealed in light of the literature on idealisations, concerns the nature of stability and structure. If stability and structure are only empirically identifiable when external factors partially determine them, then it is a matter of philosophical debate whether stability and structure are intrinsic or relational properties of a molecule. Put dif- ferently, can an isolated molecule be stable and have structure? Is the assumption of an isolated stable molecule with structure a ‘legitimate idealisation’ or an ‘outright falsehood’ (Ladyman 2008, p. 360)? This is a particularly interesting question, if one considers exam- ples of idealisations in science which are considered as false or impossible: For example, a perfectly reversible (or maximally efficient) Carnot engine is impos- sible to build in practice, and yet is considered a respectable part of the subject mat- ter of thermodynamics. On the other hand, a perpetual-motion machine of the sec- ond kind, the sole effect of which is the complete conversion of heat into work, is regarded as fundamentally impossible. What is the difference between an impossibil- ity that can be considered possible in ideal circumstances and an impossibility that remains so no matter how idealised the scenario we envisage? (Ladyman 2008, pp. 360–361) Lastly, there is a general consensus in the philosophy of chemistry literature that the man- ner in which quantum mechanics describes an isolated molecule’s structure is problematic 1 3 V. A. Seifert 28 when it comes to specifying the relation between chemistry and quantum mechanics.21 Specifically, the inability of quantum mechanics to recover a molecule’s structure is posed as a challenge to the strict Nagelian reduction of chemistry to quantum mechanics, as well as to reductive and non-reductive physicalism (see for example Hendry 2010, p. 212; Wool- ley 1998).22 Identifying that the chemical and quantum mechanical descriptions of an iso- lated molecule involve the use of idealisations, can accommodate one’s understanding of quantum mechanics’ description of molecular structure, as well as of the relation between chemistry and quantum mechanics. 22 Strict Nagelian reduction is defined here as the position which at least requires the (in principle) deriva- tion or deduction of chemistry from quantum mechanics (this is in line with Needham 2010, p. 164; Scerri 1994, p. 160). While there are different accounts of physicalism in the philosophy of science literature, physicalism (whether reductive or non-reductive) is broadly understood here as the ‘ontological position according to which the physical facts determine all the facts’ (Hendry 1999, p. 118). Philosophical implications These issues go beyond the scope of this paper. Nev- ertheless, the manner in which philosophy investigates the nature, type, and function of idealisations indicates that the identification of idealisations in chemistry and in quantum mechanics can provide novel insight into the relation between the two theories. 21 A concise presentation of this is provided by Hettema (2017, pp. 49–68). 22i Conclusion When chemistry and quantum mechanics each describe an isolated molecule, they assume that the molecule is stable and has structure. This paper showed that these assumptions are an idealisation. This is because stability and structure are relational properties when referring to a molecule that is part of an ensemble. In fact, given that the empirical iden- tification of a molecule’s stability and structure requires taking into account the environ- ment, the thermodynamic conditions and the time-range of an experiment, it follows that chemistry and quantum mechanics assume, rather than empirically verify, that an isolated molecule is stable and has structure. The paper examined these assumptions in the context of the literature on idealisations and outlined the philosophical questions that are informed by this idealisation, including the question about the relation between chemistry and quan- tum mechanics. Acknowledgements I would like to thank James Ladyman for his continuous support and advice while undertaking research for this paper. This paper was supported by the Darwin Trust of Edinburgh, the Royal Institute of Philosophy, and the European Research Council Project ‘The Metaphysical Unity of Science’, grant no. 771509. Special thanks to Nick Norman and Fred Manby of the Department of Chemistry at the University of Bristol. I would also like to thank Geoffrey Blumenthal, Robin Hendry, Tami Spector, Nicos Stylianou, Elay Shech, the reviewers of this paper, as well as the audience at various conferences in which I presented material relevant to this paper. 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https://openalex.org/W3151239091	OpenAlex	Open Science	CC-By	2,021	Understanding the impact of lumbar disc degeneration and chronic low back pain: A cross-sectional electromyographic analysis of postural strategy during predicted and unpredicted postural perturbations	Janet A. Deane	English	Spoken	10,560	19,389	The University of Manchester Research The University of Manchester Research The University of Manchester Research Citation for published version (APA): Citation for published version (APA): Deane, J. A., Macaluso, A. (Ed.), Lim, A. K. P., McGregor, A. H., & Strutton, P. H. (2021). Understanding the impact of lumbar disc degeneration and chronic low back pain: A cross-sectional electromyographic analysis of postural strategy during predicted and unpredicted postural perturbations. PLoS ONE, 16(4), Article e0249308. https://doi.org/10.1371/journal.pone.0249308 Published in: PLoS ONE Citing this paper Please note that where the full-text provided on Manchester Research Explorer is the Author Accepted Manuscrip or Proof version this may differ from the final Published version. If citing, it is advised that you check and use the publisher's definitive version. Understanding the impact of lumbar disc degeneration and chronic low back pain: A cross-sectional electromyographic analysis of postural strategy during predicted and unpredicted postural perturbations DOI: 10.1371/journal.pone.0249308 Document Version Final published version Link to publication record in Manchester Research Explorer Link to publication record in Manchester Research Explorer General rights General rights Copyright and moral rights for the publications made accessible in the Research Explorer are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Takedown policy If you believe that this document breaches copyright please refer to the University of Manchester’s Takedown Procedures [http://man.ac.uk/04Y6Bo] or contact uml.scholarlycommunications@manchester.ac.uk providing relevant details, so we can investigate your claim. Takedown policy Takedown policy If you believe that this document breaches copyright please refer to the University of Manchester’s Takedown Procedures [http://man.ac.uk/04Y6Bo] or contact uml.scholarlycommunications@manchester.ac.uk providing relevant details, so we can investigate your claim. Download date:24. Oct. 2024 PLOS ONE PLOS ONE Abstract People with chronic low back pain (LBP) exhibit changes in postural control. Stereotypical muscle activations resulting from external perturbations include anticipatory (APAs) and compensatory (CPAs) postural adjustments. The aim and objective of this study was to determine differences in postural control strategies (peak amplitude, APAs and CPAs) between symptomatic and asymptomatic adults with and without Lumbar Disc Degeneration (LDD) using surface electromyography during forward postural perturbation. Ninety-seven subjects participated in the study (mean age 50 years (SD 12)). 3T MRI was used to acquire T2 weighted images (L1-S1). LDD was determined using Pfirrmann grading. A bespoke translational platform was designed to deliver horizontal perturbations in sagittal and frontal planes. Electromyographic activity was analysed bilaterally from 8 trunk and lower limb mus- cles during four established APA and CPA epochs. A Kruskal-Wallis H test with Bonferroni correction for multiple comparisons was conducted. Four groups were identified: no LDD no pain (n = 19), LDD no pain (n = 38), LDD pain (n = 35) and no LDD pain (n = 5). There were no significant differences in age or gender between groups. The most significant difference between groups was observed during forward perturbation. In the APA and CPA phases of predictable forward perturbation there were significant differences ankle strategy between groups (p = 0.007–0.008); lateral gastrocnemius and tibialis anterior activity was higher in the LDD pain than the LDD no pain group. There were no significant differences in the unpredictable condition (p>0.05). These findings were different from the remaining groups, where significant differences in hip strategy were observed during both perturbation Editor: Andrea Macaluso, Universita degli Studi di Roma ’Foro Italico’, ITALY Editor: Andrea Macaluso, Universita degli Studi di Roma ’Foro Italico’, ITALY Received: November 20, 2020 Accepted: March 16, 2021 Published: April 1, 2021 Copyright: © 2021 Deane 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. Received: November 20, 2020 Accepted: March 16, 2021 Published: April 1, 2021 Copyright: © 2021 Deane 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. OPEN ACCESS Citation: Deane JA, Lim AKP, McGregor AH, Strutton PH (2021) Understanding the impact of lumbar disc degeneration and chronic low back pain: A cross-sectional electromyographic analysis of postural strategy during predicted and unpredicted postural perturbations. PLoS ONE 16(4): e0249308. https://doi.org/10.1371/journal. pone.0249308 Understanding the impact of lumbar disc degeneration and chronic low back pain: A cross-sectional electromyographic analysis of postural strategy during predicted and unpredicted postural perturbations Janet A. DeaneID1,2¤a¤b, Adrian K. P. Lim1,3☯¤a¤c, Alison H. McGregor1☯¤a, Paul H. Strutton1☯¤a Janet A. DeaneID1,2¤a¤b, Adrian K. P. Lim1,3☯¤a¤c, Alison H. McGregor1☯¤a, Paul H. Strutton1☯¤a Janet A. DeaneID1,2¤a¤b, Adrian K. P. Lim1,3☯¤a¤c, Alison H. McGregor1☯¤a, Paul H. Strutton1☯¤a Janet A. DeaneID H. Strutton1☯¤a a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 1 Department of Surgery and Cancer, Imperial College London, London, United Kingdom, 2 Department of Health Sciences, The University of Manchester and Manchester University NHS Foundation Trust, Manchester, United Kingdom, 3 Department of Imaging, Imperial College Healthcare NHS Trust, London, United Kingdom ☯These authors contributed equally to this work. ¤a Current address: Sackler Musculoskeletal Laboratory, Sir Michael Uren Hub, Imperial College London, London, United Kingdom ¤b Current address: School of Health Sciences, The University of Manchester, Manchester, United Kingdom ¤c Current address: Charing Cross Hospital, Hammersmith, London, United Kingdom j.deane@manchester.ac.uk Introduction Postural perturbations result from self-initiated movement or may be externally induced due to trips or slips [1], causing the centre of mass (CoM) to move close to or outside the base of support [2, 3]. This leads to system instability and subsequent loss of balance [1, 4], which may result in falls or injury if one is unable to respond effectively [3]. Translational mechanical plat- forms, designed to deliver external perturbations beneath the feet, are commonly used to examine stereotyped muscle responses, including anticipatory (APA) and compensatory adjustments (CPAs) [5–8]. APAs are described as the muscle activation that occur prior to a predicted perturbation event in order to minimise disequilibrium or falling [9–12], while CPAs are the muscle activations required to restore equilibrium following both predictable and unpredictable events [13, 14]. A recent systematic review suggests that when APAs and CPAs are evaluated in terms of muscle onset, people with chronic low back pain (LBP) exhibit significant muscle onset delay when compared with healthy controls in predicted and unpredicted postural perturbation con- ditions [15]. However, insufficient data, failure to examine both lower limb and trunk APAs and CPAs and small sample sizes, has precluded the identification of convincing differences in muscle activation or joint movement [15]. Postural perturbation strategies are usually defined in terms of muscle activation and joint movement [14]. The ‘ankle strategy’ requires the whole body to move as an inverted pendulum about the ankle joint; characterised by early activation of the dorsal ankle muscles, followed by dorsal thigh and trunk muscle activation. In contrast, the ‘hip strategy’ requires the body to move as a ‘double segment inverted with counter phase motion at the ankle and hip’ [16]; characterised by early activation of the ventral trunk and thigh muscles. In health, it has been hypothesised that the ‘ankle strategy’ and ‘hip strategy’ may be used discretely or together (‘mixed strategy’) to produce an adaptable response to postural perturbation within the sagittal plane [14]. In reality, healthy postural responses appear to represent a continuum of strategies, where strategy dominance is determined by factors including subject experience, perturbation predictability and environmental constraints [17]. Impaired postural control is often observed in those with LBP [18]. In quiet standing the ‘hip strategy’, which has been demonstrated to be more efficient in balance recovery than the ‘ankle strategy’ [19], appears restricted in people with LBP [20]. Abstract Data Availability Statement: Data generated for this manuscript are not publicly available because patient data need to be handled in accordance with the current data protection laws and ethical guidelines. Requests to access the data should be directed to the corresponding author or the NHS Health Research Authority, London Stanmore Research Ethics Committee (reference number: 13/ LO/0793, nrescommittee.london-stanmore@nhs. net). 1 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Funding: JD was funded by Versus Arthritis (grant number 20172) (https://www.versusarthritis.org). Funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. conditions (p = 0.004–0.006). Symptomatic LDD patients exhibit different electromyographic strategies to asymptomatic LDD controls. Future LBP electromyographic research should benefit from considering assessment of both lower limbs in addition to the spine. This approach could prevent underestimation of postural control deficits and guide targeted rehabilitation. Competing interests: Authors have declared that no competing interests exist. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Introduction However, this increased reli- ance on the ‘ankle strategy’ and enhanced ankle proprioceptive acuity appears to persist during more demanding balance tasks [20–22]. Impaired LBP postural control is postulated to occur as a result of pain, fear avoidance and deficits in trunk proprioception, however, the underlying mechanisms remain unclear and need to be identified in order to direct treatment [17, 21, 23]. Although some studies have demonstrated that LBP patients exhibit different peak amplitudes and muscle activations when compared with healthy controls through the evaluation of differences in lower limb or trunk postural strategy [5, 24–26], these observations are not always consistent [22, 24, 27–30]. 2 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis This reflects the heterogeneity of this patient group, for which management is challenging and remains undefined. In advance of defining a targeted rehabilitation approach, it is important to understand the impact of LBP through the examination of postural strategies in larger, more homogeneous cohorts. Lumbar Disc Degeneration (LDD) is a prevalent condition significantly associated with LBP and changes in postural control [31–37]. However, LDD is also found in asymptom- atic individuals [35, 38]. Using this cohort, the aim of this study was to investigate the differ- ences in postural control strategy between symptomatic and asymptomatic LDD patients and healthy controls through the examination of peak amplitude and integral activity of bilateral trunk and lower limb muscles, using surface electromyography (sEMG), in response to pre- dictable and unpredictable forward perturbations delivered by an in-plane translational plat- form. It was hypothesised that there would be differences between the postural control strategies adopted by symptomatic LDD patients (LDD pain) and asymptomatic controls (LDD no pain) and that these strategies would be different to those adopted by the remaining groups (no LDD pain and no LDD no pain). Participants PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 3 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis To control for bias all Modified Pfirrmann disc grades were determined by an experienced consultant radiologist, blinded to the demographics and clinical data of participants. Partici- pants were subsequently categorised into four groups including asymptomatic healthy controls (no LDD no pain (n = 19), LDD no pain (n = 38)) and symptomatic patients (LDD pain (n = 35) and no LDD pain (n = 5)). Participants were then identified as ‘LDD’ if they had mod- ified Pfirrmann grade of 6 at one or more lumbar levels [39] and as ‘LBP’ if they experienced recurrent LBP for 3 months duration (pain > 0 using a numerical rating scale (NRS)) [40]. Confounders including demographics (sex, age, weight, height and BMI) and pain (NRS) were recorded for each participant during the one-off visit to the laboratory. NRS measure- ments were recorded before the first and following the second and third perturbation to deter- mine changes in pain related to the trial. Additional self-reported clinical outcomes included the Hospital Anxiety and Depression Scale (HADS) [41] and Short Form 36, Version 2 (SF- 36) [42] to assess anxiety and depression and quality of life, respectively. To control for bias all Modified Pfirrmann disc grades were determined by an experienced consultant radiologist, blinded to the demographics and clinical data of participants. Partici- pants were subsequently categorised into four groups including asymptomatic healthy controls (no LDD no pain (n = 19), LDD no pain (n = 38)) and symptomatic patients (LDD pain (n = 35) and no LDD pain (n = 5)). Participants were then identified as ‘LDD’ if they had mod- ified Pfirrmann grade of 6 at one or more lumbar levels [39] and as ‘LBP’ if they experienced recurrent LBP for 3 months duration (pain > 0 using a numerical rating scale (NRS)) [40]. Confounders including demographics (sex, age, weight, height and BMI) and pain (NRS) were recorded for each participant during the one-off visit to the laboratory. NRS measure- ments were recorded before the first and following the second and third perturbation to deter- mine changes in pain related to the trial. Participants Additional self-reported clinical outcomes included the Hospital Anxiety and Depression Scale (HADS) [41] and Short Form 36, Version 2 (SF- 36) [42] to assess anxiety and depression and quality of life, respectively. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Participants A total of 97 participants completed the study (43 males and 54 females, mean age 50 years (SD 12), mean body mass index (BMI) 26 kg/m2 (SD 5)). Participants were recruited to this cross-sectional study through local advertising and from primary and secondary care within Imperial College NHS Trust (London, UK). Each participant provided written informed con- sent and was recruited in accordance with strict inclusion and exclusion criteria (Table 1) between September 2015 and January 2018 (NHS Health Research Authority, London Stan- more Research Ethics Committee, reference number: 13/LO/0793). A priori, a total sample size of 64 (16 per group) was estimated for a predefined 5% level of significance and 80% power based upon trunk peak amplitudes derived by Boudreau et al. [6] (GPower 3.1 Statistical Power Analyses, Dusseldorf, Germany). A 3T Verio MRI scanner (Siemens Medical Systems, Erlangen, Germany) was used to acquire supine T2 weighted sagittal lumbar spine images (L1-L5/S1) (TR = 3000 ms, TE = 92ms, 15 slices, 4mm slice width with 0.5 mm gap) from healthy controls and patients. Table 1. Inclusion and exclusion criteria. Inclusion Criteria Exclusion Criteria Asymptomatic Healthy Controls (including LDD no pain and no LDD no pain groups) • 30 years • No low back pain • No recurrent history of low back pain • No episodes of LBP lasting greater than 3 months duration • Spinal surgery • Malignancy • Spondylolisthesis • Peripheral neuropathy with loss of sensation • Systemic or spinal infection • Neurological disease or balance disorder • Disorders affecting pain perception • Significant cardiovascular or metabolic disease • Severe musculoskeletal deformity (scoliosis, osteoporosis, Paget’s disease, fracture) • Spinal surgery or major surgery within three months prior to testing • MRI contraindicated • Perturbation contraindicated Symptomatic Patients (including LDD pain and no LDD pain groups) • 30 years • Evidence of LDD without neural compression on MRI • Recurrent low back pain (central/ unilateral) of greater than 3 months duration • MRI as part of routine NHS care https://doi org/10 1371/journal pone 0249308 t001 Table 1. Inclusion and exclusion criteria. Table 1. Inclusion and exclusion criteria. Experimental procedures A bespoke perturbation platform was designed to simulate the perturbations experienced on public transport [43]. The platform accommodated feet-in-place responses from one subject standing comfortably (Fig 1). A bespoke safety harness was worn by each participant to ensure Fig 1. Perturbation platform and experimental set up. https://doi.org/10.1371/journal.pone.0249308.g001 Fig 1. Perturbation platform and experimental set up. https://doi.org/10.1371/journal.pone.0249308.g001 https://doi.org/10.1371/journal.pone.0249308.g001 4 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis safety and enable electrode placement without movement restriction. The ceiling was fitted with a zip wire, to which the harness was attached using a karabiner (Fig 1). Participants were familiarised with perturbation prior to the trial by observing forward, backward, right and left perturbations of the platform itself (40 mm in 0.2 s, average accelera- tion 1.97 m/s2). A foot template and standardardised instruction encouraged the maintenance of a consistent base of support; ‘Place your feet hip width apart and maintain this position for the duration of this experiment’. The perturbations were of sufficient magnitude to elicit com- pensatory feet-in-place responses without stepping. Subjects were instructed to stand barefoot in the centre of the perturbation platform with arms by the side. The safety harness was worn throughout the experiment. In the predictable condition participants were aware of the direction and timing of each perturbation using the advanced warning provided by an audible cue five seconds in advance of the perturbation [44, 45]. In the unpredictable condition there was no auditory cues pro- vided regarding the direction or timing of the perturbation [44, 45]. In order to limit the impact of visual cues and potential postural changes associated with visibility of perturbation graphics on the computer screen, all participants were instructed to look forward at a visual target at eye level approximately 1.5m from the platform and all perturbations were triggered with participants facing away from the computer. The conditions were presented in the same order to each subject. Subjects completed a total of three predicted and unpredicted forward, backward, left and right perturbations. Since the preliminary results indicated that the most significant difference between groups was observed during forward perturbation, this direc- tion became the focus of our investigation. Electromyography The sEMG data were acquired bilaterally from three trunk (rectus abdominis, external oblique and erector spinae) and five lower limb muscles (gluteus medius, rectus femoris, biceps femo- ris, tibialis anterior, lateral gastrocnemius) (Table 2) at 1000Hz using a sixteen channel system (Myon AG, Sonnenrain 75, 6103 Schwarzenburg Switzerland). The skin surface was cleaned with alcohol wipes and shaved if necessary. Disposable Ag/AgCl chloride self-adhesive sEMG electrodes (Neuroline 72000-S/25, Ambu Ltd. U.K., 2 cm inter-electrode distance) were posi- tioned parallel to the orientation of muscle fibres in accordance with SENIAM guidelines (http://www.seniam.org) [46] and were fixed in place with surgical tape. The raw data were bandpass filtered (Butterworth filter, 10-500Hz) and rectified using MATLAB (Mathworks, Natick, MA., U.S.A.). PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Data processing The sEMG signals were synchronised with the perturbation onset. The movement of the plat- form was assessed using an in-house constructed accelerometer (sampled at 1000Hz) attached directly to the platform. An algorithm confirmed the onset of platform perturbation using the Shewart protocol [47]. Perturbation occurred at 0ms. Differences in postural strategy were defined as a significant difference in peak sEMG amplitude following perturbation (0-350ms) or integral sEMG activation during APA and CPA time intervals defined with respect to the timing of the perturbation [1, 48]; APA1 (-250 to -100 ms), APA2 (-100 ms to +50 ms), CPA1 (+50 to 200 ms) and CPA2 (+200 to 350 ms) (Fig 2). The baseline sEMG activation was deter- mined by calculating the mean level -500ms to -450ms prior to the perturbation (0ms) [1]. All data were corrected by subtracting the baseline activation. Three times the integral of the base- line activation was subtracted from integral sEMG activation during the APA and CPA phases since each epoch represented three times the baseline window (150ms) [1]. 5 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Table 2. sEMG placement. Muscle Abbreviation Electrode Placement Rectus Abdominis RRA Electrode positioned 3cm lateral from and inferior to the umbilicus. LRA External Abdominal Oblique REO Electrode positioned superior to the ASIS and lateral to the rectus abdominis, 50% on the line between the iliac crest and the ribs. LEO Erector Spinae (Longissimus) RES Electrode positioned two finger widths lateral of the spinous processes of L1. LES Gluteus Medius RGMed Electrode positioned 50% on the line between the crista iliaca and the greater Trochanter. LGMed Rectus Femoris RRF Electrode positioned 50% on the line between the anterior spina iliaca superior to the superior part of the patella. LRF Biceps Femoris RBF Electrode positioned 50% on the line between the ischial tuberosity and the lateral epicondyle of the tibia. LBF Lateral Gastrocnemius RLGastroc Electrode positioned 50% on the line between the head of the fibula and the heel. LLGastroc Tibialis Anterior RTA Electrode positioned 33% on the line between the tip of the fibula and the tip of the medial malleolus. LTA R = Right and L = left. https://doi.org/10.1371/journal.pone.0249308.t002 https://doi.org/10.1371/journal.pone.0249308.t002 Fig 2. Representation of rectified sEMG trace. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Participants There were no significant differences in age (p = 0.28) and sex (p = 0.58) between groups and BMI was not associated with significant findings (rs = -0.20 to 0.18, p = 0.61–0.97) with one exception, right rectus abdominis in response to predicted forward perturbation (RRA APA2, rs = 0.22, p = 0.04). 100% of NRS pain scores taken prior to each trial did not change following the second or third perturbation (mean scores for the LDD pain (4 (SD 3)) and no LDD pain groups (5 (SD 2)) did not change). Data processing The original sEMG data from trunk and lower limb muscles of one healthy subject during predicted and unpredicted forward perturbation. A typical sEMG pattern averaged over three trials. 0ms represents the time of perturbation. Time scales on x axis are in ms and sEMG (mV) on the y axis. Scale (indicated bottom right) is the same for each muscle. https://doi.org/10.1371/journal.pone.0249308.g002 Fig 2. Representation of rectified sEMG trace. The original sEMG data from trunk and lower limb muscles of one healthy subject during predicted and unpredicted forward perturbation. A typical sEMG pattern averaged over three trials. 0ms represents the time of perturbation. Time scales on x axis are in ms and sEMG (mV) on the y axis. Scale (indicated bottom right) is the same for each muscle. https://doi.org/10.1371/journal.pone.0249308.g002 https://doi.org/10.1371/journal.pone.0249308.g002 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 6 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis To enable group comparisons, the peak, integral sEMG (APA1, APA2, CPA1, CPA2) and integral sum sEMG activation (APA sum (APA1+APA2) and CPA sum (CPA1+CPA2)) were normalised to the respective maximum absolute peak amplitude, maximum integral and maxi- mum integral sum of each muscle over 2 conditions and 3 perturbation trials for each muscle [24, 49]. Normalisation to the overall absolute maximum sEMG activity during postural per- turbation tasks has been previously justified since LBP patients are often not capable or willing to generate a maximum voluntary contraction [50], as was the case in our study. Preliminary observations concluded that peak amplitudes were consistently and signifi- cantly higher in the first perturbation trial than the second and third trials (p = 0.001–0.004). Therefore, the first of three perturbations was used for analysis. Statistical analysis Statistical analysis was performed using SPSS statistical package (Version, IBM SPSS statistics, IBM Corp.). Normality was determined using QQ plots, histogram and Shapiro Wilks tests. Values described in the results section are based upon mean ranks, since the distributions of integral sEMG activity for all postural adjustments were not the same (i.e. different shapes). This was assessed by visual inspection of a boxplot. Kruskal-Wallis H test was used to deter- mine if there were statistically significant differences in the mean ranks of baseline sEMG acti- vation and dependent variables (peak and integral sEMG activity (APA1, APA2, CPA1, CPA2, APA sum and CPA sum) between groups. Subsequent post-hoc analysis was performed using Dunn’s procedure [51] with a Bonferroni correction for multiple comparisons to determine group differences of the dependent variables. The effect sizes for significant comparisons of these variables were also computed (r ¼ zffiffiffi N p , where r = effect size, z = standard test statistic or z-score and N = total n for given comparison). The effects of confounding variables (age, BMI and sex) were examined using a Kruskal-Wallis H test and Spearman’s rho correlation coeffi- cient (rs). Spearman’s rho correlations were used to explore associations between muscle acti- vations (peak and integral EMG data) and LDD (LDD sum scores) [52], pain (NRS), anxiety and depression (HADS) and quality of life (SF-36). Results were considered significant at P 0.05. Missing data were excluded case wise from the analysis and was not replaced by imputed values. Data were also excluded in the event of protocol violation due to stepping reactions (1.85% of patient trials and 1.27% of healthy par- ticipant trials were excluded from the final analysis). PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Baseline activation and peak amplitude There were no significant differences in baseline sEMG activation between groups prior to predicted and unpredicted forward perturbations in the sagittal plane (p = 0.05–0.99), apart from biceps femoris (BF) during the unpredicted perturbation (p0.01). This implied that in the majority of cases, any differences observed in peak amplitude and integral sEMG activity PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 7 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis between groups were real and did not result from differences in baseline activation. There were also no significant differences in peak amplitude observed between groups following pre- dicted (p = 0.11–0.69) or unpredicted perturbations (p = 0.25–0.93). Anticipatory and compensatory postural adjustments Predicted forward perturbation. During predicted forward perturbation, integral sEMG activity was significantly different between groups; RLGastroc (APA2, H(3) = 10.838, p = 0.013), LTA (CPA2, H(3) = 9.643, p = 0.031), RRA (APA2, H(3) = 8.718, p = 0.033, CPA1, H(3) = 9.533, p = 0.023 and CPA sum, H(3) = 13.069, p = 0.004). p p Post hoc analysis revealed that during the APA phase, RLGastroc activity was higher in the LDD pain group (median rank = 55.11) than the LDD no pain group (median rank = 37.47) (RLGastroc APA2, p = 0.007, adjusted p = 0.045, medium effect size = 0.31). During the CPA phase, LTA activity was also higher in the LDD pain group (median rank = 56.70) when com- pared with the LDD no pain group (median rank = 39.74) (LTA CPA2, p = 0.008, adjusted p = 0.045, medium effect size = 0.31) (Fig 3). In contrast, the remaining groups exhibited differences in trunk muscle activation. During the APA and CPA phases, RRA activity was significantly higher in the LDD no pain (median ranks APA2 = 50.46, CPA1 = 48.68 and CPA sum = 50.65) and No LDD no pain groups (median ranks APA2 = 53.63, CPA1 = 55.26 and CPA sum = 58.11) when compared to the No Fig 3. Distribution of RLGast and LTA ranks in the APA and CPA phases of predicted forward perturbation. In the APA2 and CPA2 phases of predicted forward perturbation the LDD pain group exhibited higher RLGast and LTA mean ranks than the LDD no pain group. The group names and RLGast APA2 or LTA CPA2 (integral in arbitrary units) appear on the X and Y axis respectively. The APA2 and CPA2 phases represent 150ms (-100 to 50ms (APA2) and 200 to 350ms (CPA2) post perturbation). Significance (asterisk) and individual outliers that fell 1.5 (circles) and 3 times (stars) outside of interquartile range are indicated. https://doi.org/10.1371/journal.pone.0249308.g003 Fig 3. Distribution of RLGast and LTA ranks in the APA and CPA phases of predicted forward perturbation. In the APA2 and CPA2 phases of predicted forward perturbation the LDD pain group exhibited higher RLGast and LTA mean ranks than the LDD no pain group. The group names and RLGast APA2 or LTA CPA2 (integral in arbitrary units) appear on the X and Y axis respectively. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Anticipatory and compensatory postural adjustments The APA2 and CPA2 phases represent 150ms (-100 to 50ms (APA2) and 200 to 350ms (CPA2) post perturbation). Significance (asterisk) and individual outliers that fell 1.5 (circles) and 3 times (stars) outside of interquartile range are indicated. https://doi.org/10.1371/journal.pone.0249308.g003 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 8 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Fig 4. Distribution of RRA and LRF ranks in the APA and CPA phases of unpredicted forward perturbation. In the APA1 phase of unpredicted forward perturbation the LDD no pain group exhibited lower RRA mean ranks that the no LDD no pain group. In the CPA1 phase of the same condition, the LDD no pain group displayed significantly higher LRF mean ranks than no LDD pain group. The group names and RRA APA1 or LRF CPA1 (integral in arbitrary units) appear on the X and Y axis respectively. The APA1 and CPA1 phases represent 150ms (-250 to-100ms (APA1) and +50 to 200 ms (CPA1) post perturbation). Significance (asterisk) and individual outliers that fell 1.5 (circles) and 3 times (stars) outside of interquartile range are indicated. https://doi.org/10.1371/journal.pone.0249308.g004 Fig 4. Distribution of RRA and LRF ranks in the APA and CPA phases of unpredicted forward perturbation. In the APA1 phase of unpredicted forward perturbation the LDD no pain group exhibited lower RRA mean ranks that the no LDD no pain group. In the CPA1 phase of the same condition, the LDD no pain group displayed significantly higher LRF mean ranks than no LDD pain group. The group names and RRA APA1 or LRF CPA1 (integral in arbitrary units) appear on the X and Y axis respectively. The APA1 and CPA1 phases represent 150ms (-250 to-100ms (APA1) and +50 to 200 ms (CPA1) post perturbation). Significance (asterisk) and individual outliers that fell 1.5 (circles) and 3 times (stars) outside of interquartile range are indicated. https://doi.org/10.1371/journal.pone.0249308.g004 https://doi.org/10.1371/journal.pone.0249308.g004 https://doi.org/10.1371/journal.pone.0249308.g004 LDD pain group (median ranks APA2 = 11.25, CPA1 = 9.25 and CPA sum = 9.00) (adjusted p = 0.038 to 0.005, medium to large effect size = 0.42–0.69). Unpredicted forward perturbation. In the unpredicted forward condition, there were no differences between the LDD pain and LDD no pain groups (p>0.05). PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Peak amplitude Preliminary observations determined that there was a significant reduction in peak sEMG amplitude in all groups over three perturbations; the highest peak amplitude was observed fol- lowing the first perturbation and the lowest following the third. In the literature authors fre- quently describe computing the average or median response [1], but this can mask core differences. Therefore, in this study, the first of three perturbations was analysed to reflect the once off predictable and unpredictable perturbations that occur in daily life. However, irrespective of the predictability of the perturbation, there were no significant dif- ferences in the peak sEMG amplitudes between groups. While these findings concur with a previous LBP perturbation study [53], this is in contrast with others which found decreases and increases in trunk and lower limb muscle peak activation in response to postural perturba- tion [24, 30, 49]. The inconsistencies presented in the literature may reflect differences in experimental pro- tocol including normalisation procedures, baseline evaluation and the magnitude of perturba- tion. For example, a study which accounted for differences in baseline prior to perturbation onset and used normalisation procedures similar to that used in this current study, also found no significant differences in peak amplitudes between LBP patients and healthy controls [53]. However, in a study where significant differences in peak amplitude were found, the data did not appear to be normalised, as authors cited that LBP patients were unable to perform maxi- mum voluntary contractions to their true maximum, which may, in part, explain this differ- ence in results [30]. Anticipatory and compensatory postural adjustments However, integral sEMG activity was significantly different between the LDD no pain and remaining groups; LRF (CPA1, H (3) = 8.484, p = 0.037) (Fig 4) and RRA (APA1, H (3) = 9.714, p = 0.021). Post hoc analysis indicated that RRA activation was significantly lower in the LDD no pain group (median rank = 40.19) than the no LDD no pain group (median rank = 61.53) (RRA APA1, p = 0.006, adjusted p = 0.033, medium effect size = 0.37) and that LRF activity was significantly higher in the LDD no pain group (median rank = 51.46) than the no LDD pain group (median rank = 10.50) (LRF CPA1 (p = 0.004, adjusted p = 0.024, medium effect size = 0.44). Associations with postural strategy. In symptomatic patients, there was no direct corre- lation between APAs and CPAs and self-reported pain (NRS), LDD or depression and anxiety (P>0.05). However, in symptomatic patients, anticipatory muscle activation (RLGast APA2) correlated with SF-36 items relating to pain (‘bodily pain’ rs = 0.40, p = 0.02), physical func- tioning (‘role physical’ rs = 0.45, p = 0.006, ‘vitality’ rs = 0.55, p = 0.001) and mental health (‘emotional’ rs = 0.49, p = 0.003, ‘mental health’ rs = 0.49, p = 0.003). PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 9 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Discussion To our knowledge, this is the first time that surface translation perturbation has been used within a larger, defined LBP cohort to determine differences in postural strategy or APAs and CPAs, as determined by muscle activation. This study provides new evidence that significant differences in postural strategy occur between LDD pain and LDD no pain groups during the APA and CPA phases of postural perturbation and that these are different to those observed in non LDD groups. This is of interest since it is known that some people with LDD experience pain (LDD pain) while others do not (LDD no pain). Therefore, understanding differences in APAs and CPAs in defined LBP cohorts could help to inform patient management. Anticipatory and compensatory postural adjustments Preliminary results from our previous experiments examining predicted and unpredicted right, left and backward perturbations indicated no significant difference between LDD pain and LDD no pain groups during right or leftward perturbations (p = 0.14–0.99) and where sig- nificant differences were observed, for example, during the unpredicted backward condition, the difference in muscle activation between groups, appeared to reflect a healthy, if exaggerated response, to the study protocol, which required feet-in-place responses at high acceleration [16]. Therefore, the examination of postural strategy in response to predicted and unpredicted forward perturbations became the primary focus of this study. A recent systematic review concluded that APAs and CPAs, based upon measures of muscle onset, are delayed in response to both predicted and unpredicted perturbations in people with chronic LBP [15]. However, this review also suggested that there is no conclusive evidence that APAs and CPAs change in people with chronic LBP when measurements are based upon mus- cle activation [15]. This review not only highlighted a paucity of studies in this area but also a need for powered studies and comprehensive evaluation of trunk and lower limb muscle activation. 10 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis The strengths of this current study is that it is powered, larger than those described in the review [15] and it provides a comprehensive examination of both trunk and lower limb APAs and CPAs based upon muscle activation parameters. Using this approach, it has been possible to uncover significant differences in APAs and CPAs between groups. In the predicted forward perturbation condition, for example, lower limb muscle activations (RLGastroc APA2 and LTA CPA2) were higher in the LDD pain group when compared with the LDD no pain group. This response was different to asymptomatic controls (LDD no pain and No LDD no pain), who exhibited higher trunk muscle activations (RRA APA2, CPA1 and CPA sum) when com- pared to symptomatic LBP patients without degenerative lumbar changes (No LDD pain). In this predicted condition, the emphasis on lower limb muscle activation in the LDD pain group would appear to suggest an increased reliance on ankle APAs and CPAs in response to pos- tural perturbation. Anticipatory and compensatory postural adjustments In the absence of conclusive evidence and a paucity of studies using similar cohorts and methodologies [15], it is difficult to make direct comparison. However, in agree- ment with this study, people with chronic LBP have been reported to display significant differ- ences in TA and Gastroc activations when compared to healthy controls using similarly challenging perturbations [24, 49]. This is hypothesised to be a strategic adaptation used to avoid predictably painful, multi-segmental movement [24]. In contrast, asymptomatic controls appear to rely upon increased trunk activation (RRA) when compared to symptomatic LBP patients, which reflects previous results [49]. These findings are of interest for three reasons. Firstly, in response to predicted forward perturbation, asymptomatic healthy controls (LDD no pain and No LDD no pain) appear to favour activation of the ventral trunk, which typically contributes towards the ‘hip strategy’ [16], while the LDD pain group favour lower limb activation, which may align with the hypothesis that this group is reliant upon the ‘ankle strategy’ [16]. Although further kinematic analysis is required to verify this hypothesis, these findings are of interest since the ‘trunk stiff- ening strategy’ [54], which is associated with LBP and fear avoidance [21, 55, 56], is reliant upon the ‘ankle strategy’ [16, 21]. Secondly, studies report that people experiencing LBP dem- onstrate an ankle steered proprioceptive strategy in order to retain balance [21–23], which may explain the increased TA and Gastroc activation observed in the LDD pain group. Since this ankle steered strategy increases CoM displacement following perturbation, this finding suggests that this group could be more vulnerable to instability [5, 14]. This vulnerability is further supported by evidence that symptomatic LDD patients exhibit poor postural control during static and dynamic balance tasks [36, 37]. Finally, the observation of asymmetrical sig- nificant results as well as the apparent lack of homolateral differences has been previously doc- umented [57]. This asymmetric presentation may result from changes in axial lumbar rotation in the LDD pain group, where restricted sagittal and frontal movement, secondary to degener- ative disc changes, typically necessitates increased spinal rotation [58, 59]. Although further trunk and lower limb kinematic analysis would be required to confirm this, spinal rotation in symptomatic LDD patients is likely to have consequences for the entire kinematic chain. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Anticipatory and compensatory postural adjustments Overall, the predicted response appears in alignment with the evidence that people with LBP avoid pain provoking postures more than healthy controls in predicted conditions [60]. However, such results are in contrast to those observed in the unpredicted condition; there was no significant difference in muscle activation between the LDD pain and LDD no pain groups. Therefore, it could be concluded that it is the anticipation or prediction of the pertur- bation event (anticipatory feedforward control) that determines the ankle steered response in symptomatic LDD patients. Asymptomatic healthy controls also demonstrated strategic differences in muscle activation between predicted and unpredicted states. Asymptomatic groups (LDD no pain and no LDD no pain) exhibited higher trunk muscle activation (RRA APA1) than symptomatic patients PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 11 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis without degenerative changes (no LDD pain) in the predicted condition. However, this changed to a significantly higher lower limb reliance in the unpredicted scenario; the LDD no pain group demonstrated a higher lower limb activations (LRF CPA1) and lower trunk activa- tions (RRA APA1) than the no LDD pain and no LDD no pain group respectively. Therefore, in this study, asymptomatic healthy controls demonstrate the capacity for flexible activation of proximal muscles (RRA and LRF) depending on the context of the task. It is possible that such proximal muscle activation represents a difference in ‘hip strategy’, which is typically restricted in patients with chronic LBP [17]. Since the activation patterns of asymptomatic healthy con- trols (proximal muscle activation) are different to symptomatic patients (distal muscle activa- tion), it appears that the identification of phenotypes is possible in this cohort using this perturbation paradigm. Associations with postural strategy Correlation analysis revealed a lack of direct association between APAs and CPAs and the degree of LDD or self-reported pain, anxiety and depression experienced by symptomatic patients. However, in the predicted condition, lower limb APAs (RLGastroc APA2) were found to be directly associated with SF-36 items relating to bodily pain, mental health and physical functioning in symptomatic patients. It is noteworthy that this is the exact muscle activation parameter (RLGastroc APA2) that was observed to be significantly different between the LDD pain and LDD no pain groups in the same condition. The association between muscle activation and ‘bodily pain’ in symptomatic patients is important as it indicates the possibility that a NRS may not be sufficient in terms of determin- ing the true impact of LBP. In fact, it seems that when patients are invited to report their pain experience over a longer time period (four weeks) and to reflect upon how much pain inter- feres with their lives that it becomes feasible to uncover the association between pain and pos- tural strategy. It is possible that this correlation reflects a change in the ‘central set’ or the preparatory state of the CNS in response to predictable perturbations, proposed to result from an increased arousal of the nervous system due to pain or pain related fear avoidance move- ment in patients with active LBP [28]. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 Acknowledgments We would like to acknowledge David Buckwell, Dr Clement Favier, Dr Megan Sperry and Dr Andrew Phillips for their expert advice and support in designing and developing the perturba- tion platform used in this study. Thanks to Dr Djordje Brujic for his generous support with the development and creation of bespoke MATLAB scripts and to Lesley Honeyfield (Lead Research Radiographer, Charing Cross Hospital, Imperial College Healthcare NHS Trust, London) for assisting with imaging optimisation and the acquisition of MR images. Finally, many thanks to the participants in this study and to the National Institute for Health Research (NIHR) for supporting recruitment. Conclusions LDD patients with chronic pain exhibit different postural strategies from asymptomatic LDD controls. These strategies are associated with task predictability, different to those exhibited between other groups and are not restricted to the lumbar spine. Acknowledgement of such maladaptive strategies, through the comprehensive evaluation of the spine in addition to the lower limbs, may prevent future underestimation of deficits in postural control and guide tar- geted rehabilitation. Further kinematic analysis will be required to understand how LBP impacts upon movement selection strategy in LDD patients. Limitations In this study a priori testing confirmed that a total sample size of 64 subjects (16 per group) would be required to obtain sufficient power. This estimate was overachieved by recruiting 97 subjects with at least 19 subjects per group (no LDD no pain = 19, LDD no pain = 38, LDD pain = 35). However, despite ongoing recruitment, it was not possible to recruit more than 5 subjects to the ‘no LDD pain’ group, indicating the rarity of symptomatic adults (>30 years) without degenerative lumbar change. Therefore, results pertaining to this group should be interpreted with caution and are not generalisable. It is accepted that the differences in muscle activation strategy observed in this study do not reflect differences in kinematic strategy, nor do they provide information regarding co-con- traction. Therefore, future analysis will be required to elucidate this. Although it could be con- sidered a limitation that the ‘unpredictable’ condition in this study potentially underestimated differences, as participants had their eyes open throughout the trial, patient and public involve- ment in advance of the trial determined that patients did not feel safe and would be less likely to consent if their eyes were shut during the trial. Therefore, a suitable compromise was reached, such that participants focussed on a visual target and did not receive visual prompts or audible cues. It is acknowledged that causation cannot be implied from this study. Therefore, it is not possible to confirm whether differences in postural control result from degenerative change, changes in reflex response modulation by higher centres, changes within the cerebral cortex (or subcortical structures) or represent difficulty in interpreting feedback. Further longitudinal research will improve understanding of these processes. Clinical relevance The findings of this current study suggest that differences in postural responses between LDD pain and LDD no pain groups are similar to the differences found between non-specific LBP and healthy subjects and may prove modifiable. While motor retraining has the capacity to change postural strategy [61–64], understanding specific impairments in motor control may assist the advancement of targeted treatment design and improve patient outcomes. It has been demonstrated in this study that the response to postural perturbation depends on the predictability of the task. In the unpredictable condition, LDD pain patients moved similarly to the LDD no pain group because they could not predict the event. However, in pre- dicted conditions LDD pain patients use maladaptive strategies, which have been found to be associated with pain and quality of life in this current study, which could increase the likeli- hood of falls or injury in this group by increasing susceptibility to centre of mass displacement secondary to reliance on the ankle strategy. The observation that changes in postural strategy do not occur exclusively in the trunk and that changes may be observed in the lower limbs, suggests that a comprehensive assessment of the trunk and bilateral lower limbs is important. This approach may avoid potential underesti- mation of motor control impairment and treatment efficacy in future [65–67]. PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 12 / 18 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Formal analysis: Janet A. Deane, Adrian K. P. Lim, Paul H. Strutton. Conceptualization: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Data curation: Janet A. Deane. Formal analysis: Janet A. Deane, Adrian K. P. Lim, Paul H. Strutton. Author Contributions Conceptualization: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Data curation: Janet A. Deane. Formal analysis: Janet A. Deane, Adrian K. P. Lim, Paul H. Strutton. 13 / 18 PLOS ONE \| https://doi.org/10.1371/journal.pone.0249308 April 1, 2021 PLOS ONE Understanding the impact of lumbar disc degeneration and chronic low back pain: An electromyographic analysis Funding acquisition: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Investigation: Janet A. Deane. Methodology: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Project administration: Janet A. Deane. Resources: Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Software: Paul H. Strutton. Supervision: Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Validation: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Visualization: Janet A. Deane. Funding acquisition: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Funding acquisition: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Investigation: Janet A. Deane. Methodology: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. P j d i i i J A D Methodology: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Project administration: Janet A. Deane. Resources: Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Supervision: Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Validation: Janet A. Deane, Adrian K. P. Lim, Alison H. McGregor, Paul H. Strutton. Visualization: Janet A. Deane. Visualization: Janet A. Deane. Writing – original draft: Janet A. Deane. Writing – review & editing: Janet A. Deane, Adrian K. P. Lim, Alison H. 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https://openalex.org/W2975586688_1	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	5,753	11,051	fa Vida y ta Niotaxia Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano LAS CUATRO CATEGORÍAS TÉCNICO MATERIALES DEL ARTE RUPESTRE PERUANO MATERIAL AND TECHNICAL FOUR CATEGORIES OF PERUVIAN ROCK ART Gori Tumi Echevarría López RESUMEN Haciendo un balance histórico de los estudios rupestres peruanos se puede concluir que, en la actualidad, la forma como se estima esta evidencia cultural en el Perú es completamente arcaica, supervalorando únicamente algunos componentes materiales mínimos de esta evidencia y relegando completamente otros componentes asociados importantes. Esta apreciación fragmentaria se basa principalmente en el desconocimiento de la naturaleza material del arte rupestre yen el poco interés científico en establecer una definición técnica en base a categorías formales con valor epistemológico. Como vamos a exponer en este artículo, la incorporación de categorías definitorias del arte rupestre peruano, desde una perspectiva arqueológica, tiene implicancias en la forma como este material es entendido, registrado y estudiado; siendo claves para su conservación, protección, yen especial, para dotar de valor social a esta importante evidencia cultural. Palabras clave: ABSTRACT Making a historical record of Peruvian rock studies it can be concluded that, at present, the way how this cultural evidence in Pera is estimated is completely archaic, overvaluing only some minimal material components of Mis evidence and completely relegating other important associated components. This fragmentary assessment is mainly based on the lack of awareness of materia Is for rock art and the finte scientific interest in establishing a technical definition based on formal categories with epistemological value. As we will explain in this article, the incorporation of defining categories of Peruvian rock an, from an archaeoloeical perspective has implications for how this material is understood, recorded and studied; being the keys for conservation, protection, and in particular to provide social value lo this important cultural evidence. Key words: INTRODUCCIÓN había reconocido muchas marcas sobre rocas como signos culturales, y su uso durante tiempos históricos, especialmente durante el Tahuantinsuyu (1470— 1533), es atestiguado por varios cronistas de la colonia como bien ha concluido Raúl Porras Barrenechea en su clásico estudio "Quipu y Quilca" de 1947 (Porras 1963). Porras identifica claramente, como un hecho histórico, la existencia de dos variaciones gráficas simbólicas o escriturales para las sociedades andinas, expresadas individualmente con los vocablos nativos quipu y quilco. Para Rivero y Tschudi (ob. cit.) pero especialmente para Porras (ob. cit.) la dualidad de este registro gráfico simbólico es contundente y se expresa en un sin número de notables fuentes históricas relacionadas. Aunque es bastante evidente que los Las primeras definiciones del "arte rupestre" en el Perú han sido totalmente transplantadas del acervo europeo, donde el arte rupestre o "arte parietal" fue registrado y estudiado sistemáticamente desde el siglo XIX (Álvarez 2006). Los términos asociados a estas definiciones, como "pictografias" o "petroglifos" incluidos dentro del concepto genérico de arte rupestre se han usado en el Perú desde el siglo XX, aunque las primeras descripciones de estos materiales, hechos por intelectuales y viajeros ilustrados del siglo XIX, han incluido frecuentemente conceptos relacionados con el de "jeroglifos" o simplemente "inscripciones" (Rivero y Tschucli 1851 [1958]: 73, Hutchinson 1873: 174) No obstante esto la tradición peruanista ya 'Arqueólogo. Universidad NacionalMayorde San Marcos. Presidente dela Asociación Peruana deArte Rupestre (APAR). Lima-Perú. Correoelectránico:goritumi@gmait con? Recibido: 18/10/2014 Aceptado: 28/11/2014 ISSN 2312-9115; 2014(2)VoL 111 :38-47 38 La Vida y la Mistada Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano quipus constituyen hasta hoy una variación etnográfica de los sistemas de registros andinos, las quilcas no habían sido estimadas seriamente hasta los estudios de Porras quien demuestra que existe una relación vinculante directa entre el significado del término y el hecho que este implica: la pictografía o la escritura. De acuerdo a los estudios de Porras, las referencias a pictografías que se relacionan a los vocablos guaca o quellca provenientes de las lenguas peruanas Quechua o Aymara respectivamente, son utilizadas de forma sistemática e inequívoca en los documentos históricos de la colonia temprana, como las crónicas de Titu Cusi Yupanqui de 1560, la de Huaman Poma de Ayala (escrita probablemente entre 1567 y 1615 [Porras 1999:53]), o la crónica de Montesinos (siglo XVII); pero especialmente en los vocabularios lingüísticos como el "Lexicón' de Fray Domingo de Santo Tomás de 1560, la "Doctrina Cristiana" de 1584, el vocabulario del Padre Diego González de Holguín de 1608, el vocabulario de Francisco del Canto de 1614, o el Diccionario de Torres Rubio de 1619 (Porras ob. cit.) Es evidente entonces que el término quilca se relaciona técnicamente a un determinado tipo de actividad y hecho físico; y es el mismo Porras quien concluye al respecto: "No cabe, pues, duda de que los indios prehispánicos del Perú tuvieron una palabra especial para denominar los signos escritos y que esta palabra se aplicó más tarde por analogía, a la escritura española y al papel que no conocieron los Incas. Esto no quiere decir que ambos sistemas de escritura fuesen iguales sino que se utilizó en quechua el término lingüístico más próximo o semejante para calificar el invento occidental. También se concluye que la palabra quilca estaba unida a la idea no sólo de signos gráficos sino de signos coloreados o de dibujos o pinturas. La voz Quilca trae de por sí aparejada una representación cromática. Escribir, dibujar, pintar se expresan por una sola palabra: guillen, y esta misma palabra sirve para expresar el lienzo, la tabla o el objeto pintado. Cabe, pues, usar la palabra guaca como sinónimo de pictografía en el concepto de los etnólogos modernos?' (Porras 1963: 112). Lo establecido por Porras es sin duda notable, sin embargo este autor no asoció directamente el concepto guaca con el de las pinturas rupestres, aunque en su estudio sí alude indirectamente a los petroglifos sugiriendo una relación casual respecto de la existencia de algún tipo de "escritura" antigua en el Perú por la conspicua presencia de este material cultural en los desagregado y es básicamente un concepto literal, histórico, es claro que se había hecho el primer acercamiento serio para la integración histórica material de este término. "Quilcas", no obstante, no define al arte rupestre por sí mismo, es solo el nombre que los antiguos peruanos le otorgaron a ese fenómeno en el momento en que los españoles arribaron al Tahuantinsuyu (Perú); las poblaciones Quechua o Aymara hablantes las llamaron quilcas aún cuando ellos supieron que las pinturas o los petroglifos no fueron de su propio tiempo. El uso del término fue probablemente asignado al arte rupestre por muchos años, incluso antes de que los gobernantes del Tahuantinsuyu (Incas) gobernaran nuestro país. Se desconoce aún cuando se introdujo el término quilca, pero la lengua Quechua se originó al menos 2000 años antes del presente en la costa central del Perú (Shady 2000) Más allá de Porras, el primer investigador que logra reconocer una relación contundente entre el concepto quilca y pinturas rupestres es Javier Pulgar Vidal. Pulgar Vidal desconocía cualquier argumento historicista cuando en la década de 1930 establece la primera asociación técnica entre el termino quilca y la existencia de arte rupestre. Para esto Pulgar Vidal identificó primero el significado de la palabra compuesta "Quilla-Rurni" (el nombre de una roca sobre un cerro en la cuenca del río Higueras en Huanuco) descubriendo que el término quilla era una derivación española de una palabra en lengua nativa "Cauqui" cuya acepción literal es "grafía" o "signo gráfico"; revelación que fue seguida con la exploración de la roca en 1936 encontrando que esta era el soporte de más de "doscientos signos ideográficos" (Pulgar 1946: 12), es decir pinturas rupestres, A partir de este hallazgo Javier Pulgar elaboró la más importante hipótesis para el descubrimiento de sitios con arte rupestre en el Perú, estableciendo una relación directa entre el topónimo quilca, en todas sus variantes lingüísticas, y la presencia de pinturas o pictografías sobre roca. Aunque, como hemos visto, la identificación física y el hallazgo de sitios con arte rupestre ISSN 2312-9115; 2014(2)Vol. 111 :38-47 39 Ea Vida y ea Niotaxia Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano alcanzaron niveles sobresalientes, la estimación material de las guacas, como un artefacto arqueológico definido quedo subordinada para la propia arqueología peruana y este material, identificado corrientemente como "arte rupestre", no fue considerado primordial en los estudios arqueológicos nacionales cuyas investigaciones relacionadas lamentablemente no siguieron las tendencias de localización y reconocimiento iniciadas por Javier Pulgar Vidal y sus alumnos de la Universidad de San Marcos. Es importante notar que a partir de la década de 1960 los estudios rupestres fueron relegados frente a los notables avances de la arqueología peruana respecto de otros materiales, cerámica o arquitectura, o frente a otros problemas culturales (Echevarría 2008). No obstante esta situación hay que destacar que variadas investigaciones arqueológicas significaron para el arte rupestre, por este mismo vínculo o asociación asimétrica, un avance claro en el conocimiento de este material respecto del establecimiento de contextos arqueológicos o culturales, especialmente para las épocas más tempranas del poblamiento andino, y los estudios de Cardich en Lauricocha (1964), Neyra en Sumbay (1968), Muelle en Toquepala (1969), o Linares Málaga en una perspectiva general para Arequipa (1973) son ejemplos en este sentido. Hay que mencionar también que se intentaron aproximaciones directas, y la más importante contribución la constituye el análisis formal comparativo de Toribio Mejía Xesspe (1968), quien establece arte rupestre culturalmente asociado, en este caso arte rupestre Chalán. Aunque estos estudios son importantes, debemos reconocer que el arte rupestre, las quilcas en la acepción más extendida de Pulgar Vidal, ha continuado siendo un material sin definición propia cuyos atributos particulares han sido generalmente obviados a favor de la supervaloración de la imagen graficada en la roca, siendo muy pocos los investigadores que han reconocido otros atributos materiales relevantes más allá de esta misma imagen, o que hayan llevado a cabo un enfoque de investigación que aborde el material como una variable principal en un estudio arqueológico convencional, tal como hizo Mejía Xesspe. A este respecto el adelanto más resaltante ha sido sin duda el trabajo sostenido de Eloy Linares Málaga quien, desde finales de la década de 1950, emprendió estudios arqueológicos utilizando al arte rupestre como una variable dominante ya sea en sitios como Toro Muerto (Linares 1960, 1974), o evaluando técnicamente el arte rupestre de Arequipa (Linares 1973,1988). A comienzos de los años setenta Eloy Linares Málaga va a establecer de forma notable la tipología variantes ("pictografias", "petroglifos", "arte rupestre mobiliar" y "geoglifos") las que hasta hoy constituyen la principal base ordenadora de este material arqueológico peruano. La distinción, explícitamente tipológica, con fines de investigación material es un desarrollo técnico en la comprensión del valor cultural de las Micas del Perú, que constituye una contribución importantísima para la arqueología nacional y que obviamente ha permitido desarrollar investigaciones posteriores derivadas Sin embargo, aunque Eloy Linares Málaga trató al arte rupestre como un objeto arqueológico regular, incluyéndolo corrientemente en ensamblajes arqueológicos complejos (Linares 1973, 1988, 1999), este material quedo siempre supeditado al criterio técnico del autor respecto de sus características intrínsecas, las que son independientes de su distinción tipológica convencional o del tipo de tratamiento regular como variable científica. Esto es interesante porque, salvo Linares Málaga, muy pocos investigadores han logrado desarrollar estudios favoreciendo variables provenientes de las propiedades intrínsecas del material o han considerado a las guacas como un artefacto en sentido expreso. Después de Linares Málaga, las contribuciones en los estudios rupestres han sido muy numerosas, diversificadas y genéricas y es difícil evaluarlas en detalle aquí, aunque ya hemos advertido (Echevarría 2008) claras tendencias hacia la sistematización científica usando principalmente diferentes aproximaciones metodologías, en variables materiales directas (Morales 1993; Echevarría 2003, 2004) o en otras variables relacionadas, como etnográficas (Bueno 2006) espaciales (Nieves 2006), o visuales (Campana 2007) entre otras; sin embargo es cierto también que la falta de una perspectiva técnica en la identificación de las propiedades intrínsecas del arte rupestre peruano ha generado, más que investigación pura, registros fragmentados en el mejor de los casos basados en algunas propiedades particulares de estos materiales, sea la figuración, la técnica, o el tipo petrológico del soporte; siendo el registro de Antonio Núñez Jiménez (1986) una contribución de este tipo. En los últimos veinte arios la ignorancia en el tratamiento de este material arqueológico ha producido relaciones irregulares de sitios con arte rupestre, basados principalmente en referencias secundarias (Ravines 1986) y en el peor de los casos en referencias falsas y dudosas, en el uso de bibliografía no corroborada o inexistente, y en la supervaloración de la imagen figurada (Hostnig 2003); aún así, algunas de estas documentaciones son apócrifamente llamadas "inventarlos" por sus propios autores. La existencia de documentación fragmentaria o apócrifa tipifica en parte el nivel académico actual de 1SSN 2312-9115; 2014(2)17(1111:38-47 40 — .ea Vida y ea Niotwila Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano carencia técnica respecto al reconocimiento y tratamiento del arte rupestre peruano. Esto es potencialmente peligroso hacia el material puesto que genera una estimación negativa del valor real de las quilcas, como un objeto cultural y como una reliquia arqueológica con propiedades particulares complejas que van más allá de su imagen figurada. La falta de propuestas más estandarizadas sobre el tratamiento técnico del arte rupestre indica que los estudios en este material han seguido principalmente parámetros generalizados de apreciación artística e interpretativa, prosiguiendo desarrollos cada vez menos convencionales y sin valor científico. Hasta año 2007, no obstante, la recientemente fundada Asociación Peruana de ArteRupestre ca para (APAR) propone el primer Código de Éti visitas a sitios arqueológicos con arte rupestre que incluye una explicita definición de este material, al que considera un objeto arqueológico en sentido expreso. Este Código de Ética propone premisas conceptuales dotando al arte rupestre de características propias particulares entre las que se incluyen la "figuración", el "soporte" y el "entorno paisajístico"; las que conforman en conjunto el "sitio arqueológico de arte rupestre". Esta es probablemente la primera vez en la historia de los estudios de este material cultural en el Perú en el que se intenta establecer una definición material explicita del arte rupestre. No obstante este acercamiento, considero que la definición de APAR es todavía insuficiente para poder cubrir cabalmente las propiedades particulares de este artefacto, aunque es evidente que se ha tomado en cuenta genéricamente la mayoría de sus propiedades básicas. En la actualidad es imperioso desarrollar una conceptualización lógica explicita de estas propiedades y justificar su valor en referencia a algunos aspectos cruciales de su uso en actividades académico culturales relacionadas, como son el registro, la conservación o la investigación científica. Pienso que una definición del tipo que se intenta realizar, redunda únicamente en la inclusión definitiva de las quilcas o el arte rupestre dentro de los bienes patrimoniales más importantes del Perú, entre las que estuvo relegada por falta de perspectivas técnicas claras, como ya vimos más atrás. Las categorías Decir que el arte rupestre es un artefacto arqueológico en el Perú no es una definición ilusoria o un capricho nominal, las leyes peruanas protegen textualmente al arte rupestre al considerarlo en forma general dentro de sus bienes arqueológicos inmuebles (ley 28296). Sin embargo, aunque el estatus legal y de protección estatal regulariza la inclusión del arte rupestre dentro de sus bienes patrimoniales, las leyes no establecen las características particulares de los objetos arqueológicos debido a extrema amplitud o variabilidad material; por lo que es claro que el establecimiento de las particularidades intrínsecas de los materiales arqueológicos, como las guacas, depende fundamentalmente de los investigadores que proponen estas cualidades para sus propios fines. En este sentido una definición regular, sin indicaciones subjetivas, es necesaria. De acuerdo a Robert Bednarik, "Una científica definición de arte rupestre (...) es que este consiste de marcas que ocurren en la superficie de las rocas que fueron "intencionalmente" producidas por miembros del género Horno (me. marcas antrópicas), las que son detectables por las facultades sensoriales "normales", y que son externalizaciones de conceptos meditados de una "conciencia" advertida de la realidad que percibe" (Bednarik 2007. Traducción mía). Como se puede ver los criterios técnicos científicos para el reconocimiento del arte rupestre pasan más por la definición de su naturaleza físico cultural -deliberada hechura humana-, que por su significado implícito en algún nivel de aprensión sociocultural, y esto es crucial para su entendimiento adecuado. El valor del arte rupestre, como objeto o artefacto cultural, es independiente al valor de la apreciación subjetiva que se pueda tener sobre una cualidad particular del mismo objeto, como la calidad del dibujo o de la pintura que presenta; ya que toda marca cultural o figuración en la roca tiene su propia carga social significativa que le es particular e intrínseca. A partir de aquí, y siguiendo los parámetros básicos de identificación regular del objeto arqueológico llamado genéricamente arte rupestre -quilcas- propuestos por la Asociación Peruana de Arte Rupestre (APAR 2007), estimo que debemos considerar al menos cuatro características intrínsecas mínimas del material rupestre para fines de su uso y tratamiento regular en el Perú, estas características convertidas en categorías formales son: la imagen figurada o motivo, el soporte, el entorno inmediato, y el paisaje. La imagen figurada o el motivo La imagen figurada o el motivo es probablemente la más importante propiedad del arte rupestre y constituye el eje del reconocimiento material de este objeto arqueológico. Independientemente de su manufactura, de su escala, o de su ubicación, la imagen figurada otorga un valor cultural al material físico, la roca, al dotarlo de representatividad gráfica. La imagen es un ardid de graficación cultural y es 1SSN 2312-9115; 20140) Vol. 111:38.47 fa Vida y. ta Ajotada Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano hombre mediante recursos físicos materiales, ya sea utilizando técnicas reductivas (petroglifos) o aditivos (p.e. pinturas); debiendo ser reconocida por el hombre de forma visual simple (Figura 1). Para que exista arte rupestre debe haber un motivo inscrito, y este reconocimiento debe poder ser descrito, cuantificado, o ser sujeto de medición. atributo rupestre debe ser identificado dentro de parámetros situacionales respetando su integridad volumétrica y espacial; a lo que debe sumarse la estimación de sus propiedades, que debe estar resuelta en las variables físicas que le son constituyentes más allá de la escala, como sus aspectos mineralógicos, todos cuantificables. El soporte El soporte es la base de la figuración, este debe ser necesariamente roca de cualquier tipo y puede estar ubicado en cualquier contexto de exposición geológica (Figura 2). Dado que el soporte sirve de base para la representación figurada sufre la transformación física de su estructura al ser afectada por esta figuración; esto condiciona el material rupestre a las características físico químicas del soporte, es decir de la roca, por lo que el soporte constituye el sustrato físico que da integridad material al objeto arqueológico. Imagen figurada y soporte son condicionales y necesariamente se implican respecto de su materia, conformando una unidad física. Siendo físico el soporte debe ser también cuantificable, es decir sujeto de medición. Aunque la escala del soporte puede variar desde pequeñas piedras muebles, lajas, cantos rodados, remanentes aluviónicos, o verdaderos farallones de roca expuestos sin discontinuidad evidente, este El entorno inmediato Independientemente de la imagen figurada y del soporte, que conforman una unidad física, el entorno inmediato es una propiedad externa del artefacto rupestre puesto que este sirve de base inicial en la ubicación referencial del material arqueológico, con el que forma una unidad contextual primaria de correlación arqueológica. Esta referencia es consustancial al hecho que la producción de arte rupestre implica un entorno circunscrito que es transformado mientras se produce el arte rupestre, tal como se transforma el entorno de la manufactura de cualquier instrumental litico, con la diferencia que el arte rupestre es generalmente inmueble y se realiza en una locación fija. La estimación del valor del entorno inmediato del arte rupestre peruano ya se ha hecho desde la década de 1950 cuando el Dr. Eloy Linares Málaga hizo la primera sugerencia controlada para el uso de Figura 1. El ino va cn un priroylifo del sitio arqueológico Checta. Lima, Perú (imagen digitalmente enfatizada). ISSN 2312-9115; 2014(2) Vol. Ill :38-47 42 - £a Vida la »atada Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano Figura 2. El soporte en un petroglifo del o arqueológico Checta. El paisaje El paisaje en el arte rupestre debe considerarse, como la categoría anterior, un atributo externo pero un valor intrínseco del objeto, puesto que constituye su soporte situacional físico e implica los contextos de afectación cultural y natural que condicionan la existencia y la conservación del mismo; entre los que se encuentran los agentes antrópicos presentes y los condicionantes físico naturales asociados, como son la naturaleza geológica y medioambiental que la afectan. El paisaje, como un componente externo de los sitios y materiales arqueológicos, es por lo tanto un elemento de su constitución con valores referenciales e implicancias culturales. Como todos los atributos técnico-materiales del arte rupestre, el paisaje involucra una escala física cuantificable que en este caso se encuentra condicionada por la naturaleza histórica de formación del sitio y su imagen general reconocida. Por lo tanto, el paisaje en el arte rupestre está constituido por los medios circundantes que afectan el artefacto rupestre, soporte, medio ambiente y cultura, que han determinado su formación como yacimiento arqueológico (agentes tafonómicos mediatos), y por la imagen actual, "natural", del sitio (Figura 4). artefactos liticos en la producción de arte rupestre en sitio de Toro Muerto (Linares Málaga 1960), y este tipo de estudios se han llevado a cabo después en otras regiones andinas, como en Bolivia (Querejazu 2001), lo que confirma una relación directa entre el material rupestre, en el sentido del artefacto, y el entorno inmediato asociado. La distancia regular del entorno debe estimarse de acuerdo a las características de ubicación del sitio con arte rupestre, y su potencial arqueológico. En el también llamado sitio de "Toro Muerto" en la cuenca del río Mizque (Bolivia), Robet Bednarik recupero artefactos fabricados para hacer arte rupestre ("mur-e") asociados al entorno inmediato de los petroglifos, los que le sirvieron después para datar estas evidencias (Bednarik 1998). Aunque la cueva de "Toro Muerto" contiene un radio externo de más de cincuenta metros con materiales arqueológicos asociados; otros sitios, como "Checta" o "El Alto de las Guitarras" que contienen piedras con petroglifos en áreas muy reducidas, deben incluir necesariamente radios de áreas menores cuya estimación deben ser evaluada adecuadamente para poder reconocer el entorno donde pueda hallarse evidencia cultural significativa, ya sea para la producción de arte rupestre o la inferencia de actividades y/o relaciones culturales relevantes vinculadas a este material (Figura 3). ¡SS?'! 2312-91.15; 20140) Vol. 111:3847 43 La Vida y la Nietoda Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano Figura 3. Un ejemplo arbitrario del entorno inmediato en un petroglifo del sitio arqueológico Gaceta. Figura 4. Una vista del paisaje en el sitio arqueológico Checta Las implicancias Como se puede colegir, las implicancias del reconocimiento de las cuatro categorías técnico — materiales del arte rupestre peruano afectan inmediatamente la forma como este objeto es percibido, especialmente para fines de tratamiento material y estudios controlados; y esta es precisamente la intención de la propuesta, el cambio en la comprensión conceptual del arte rupestre peruano a partir de su identificación material primaria como un ISSN 2312-9115; 2014(2) vol. III :3847 artefacto arqueológico convencional, con propiedades particulares cuantificables. Aunque ya he mencionado más arriba que los atributos particulares de los materiales arqueológicos son generalmente propuestos por los investigadores que estudian estos materiales, la definición de estas propiedades básicas debería afectar el desarrollo de los estudios rupestres en un sentido material expreso. Vamos a comentar someramente algunas de estas implicancias relacionadas al registro, la conservación y la investigación de este artefacto. £a Vicia fl ea Niatosiit Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano Registro La consideración expresa de las cuatro categorías materiales del arte rupestre debe afectar definitivamente el reconocimiento de este artefacto, condicionado la inclusión en el registro de aspectos no tradicionalmente tomados en cuenta en la identificación de los componentes particulares de este objeto, como el soporte o el paisaje, con el mismo valor normalizado para fines de documentación básica. Un buen reconocimiento, al menos en algunos aspectos físicos relevantes, de un material cultural, incrementa notablemente la utilidad y la importancia de los registros ya sea para investigación o fines más genéricos, como inventarios, catastros, o mapas, los que son usados corrientemente en estudios arqueológicos convencionales. Aunque el registro técnico del arte rupestre presenta potencialmente variables ilimitadas, la inclusión de estas cuatro propiedades materiales como rubros estandarizados para una observación genérica busca identificar el objeto, dentro de su propia naturaleza fisica, como un corpus indivisible para su uso científico. relación se aplica a la pérdida o destrucción del soporte, del entorno o del paisaje asociado; como he dicho cada propiedad incluye valores consustanciales a la naturaleza del arte rupestre que le son esenciales e insustituibles La conservación integral de las propiedades físicas del arte rupestre permite además establecer parámetros regulares de preservación que pueden ser usados científicamente para la comprensión de variados aspectos relacionados a esta evidencia, como los procesos tafonómicos que la afectan, los contextos de producción o manufactura rupestre, los contextos de significación cultural y cualquier otro aspecto contextual asociado, comúnmente estudiado por la arqueología. Investigación La definición de las propiedades particulares del arte rupestre amplía considerablemente los alcances del estudio en este material arqueológico al incluir otros aspectos físicos del objeto, no usualmente tomados en cuenta, como variables de una investigación convencional con perspectivas científicas. En arqueología esto permite abstraer el material a aspectos analíticos básicos relacionados, como el establecimiento de contextos arqueológicos, asociación cultural, cronología y ubicación; los que pueden establecerse sin reducir el objeto rupestre a variables singulares o exclusivas. Esta inclusión, epistemológicamente controlada, permite esencialmente dotar de un valor científico extendido a la investigación rupestre al habilitar nuevos parámetros de análisis lógicos en el procesamiento de la información material obtenida y al permitir la inclusión de otras ciencias, como geomorfología, física, química, etc., en estos mismos estudios. Este cambio, en la lógica y en la colaboración científica, es positivo e irreversible y constituye de hecho el más grande avance en la investigación rupestre contemporánea (Bednarik, 2007: 12). La incorporación de más variables controladas a la investigación científica del arte rupestre brinda oportunidades ilimitadas al conocimiento de este artefacto cultural, dotándolo de un valor complejo que puede ser reconocido independiente por sus propias cualidades intrínsecas, sea su locación, su soporte, o su imagen, etc. Al complejizax su constitución, por el reconocimiento de estas propiedades, las guacas puede inscribirse, mediante la investigación, en los anales de correspondencia histórica, ajenos de las simples apreciaciones estéticas superlativas, y vinculados positivamente a los agentes sociales de los que han derivado. Conservación y protección El reconocimiento de las propiedades materiales del arte rupestre tiene una importancia fundamental en la conservación y protección de esta evidencia cultural, pues permite establecer con contundencia los límites y alcances de cualquier intervención directa en el objeto ya sea para su estudio o tratamiento científico; y esto incluye la conservación del motivo, del soporte, del entorno inmediato y del paisaje que enmarca el sitio. Las cualidades designadas, independientemente de su variabilidad, son genéricas y obligan a considerar una preservación integral que no sobrevalore ningún aspecto específico de los artefactos y/o sitios con arte rupestre en detrimento de otros; en este sentido es evidente que la conservación de una propiedad particular del arte rupestre puede acarrear la destrucción de otras propiedades de la misma importancia. Por ejemplo la supervaloración del motivo sobre el soporte que puede inducir a la descontextualización de la imagen figurada mediante el retiro y traslado del sustrato que la soporta. Al perder una de sus propiedades por la supervaloración, •o conservación diferenciada, de alguna otra propiedad específica (que generalmente está centrada en la imagen figurada), el arte rupestre queda destruido y pierde probablemente todos los valores culturales con los fue dotado originalmente, además de su significado y sentido social. Esto es como preservar sólo una copia dibujada de la imagen original del artefacto rupestre, o como conservar únicamente la pintura decorativa de una vasija cerámica; y esta 1SSN 2312-9115; 2014(2)Vol. ¡11:38-47 45 .ext 'Vida ta Mistada Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano CONCLUSIONES Debo recalcar que esta es una propuesta inicial para el tratamiento técnico del arte rupestre peruano, cuyas implicancias alcanzan aspectos culturales relacionados a estos bienes arqueológicos como son el registro, la conservación y la investigación de estos mismos bienes. Considero que las cuatro categorías propuestas, imagen figurada o motivo, soporte, entorno inmediato y paisaje, constituyen características o propiedades intrínsecas mínimas de estos materiales, cuyo uso debe hacerse a partir del conocimiento explícito de estas propiedades, y esta es la intención del discurso. El valor epistemológico del conocimiento se basa en la cabal comprensión de las categorías conceptuales que son usadas para desarrollas estos conocimientos a partir de los hechos de la naturaleza; estas categorías no pueden estar implícitas si lo que se quiere es orientar el tratamiento conciente e intelectual de los bienes culturales peruanos como las quileas. Julio C. Tello afirmaba en 1922, que: "La investigación o escudriñamiento consiste en la aplicación sistemática de los métodos de la ciencia, en el dominio de lo desconocido, por aquel que posee espíritu científico." (Tello 1965). Este espíritu es patente en los pioneros en el estudio del arte rupestre peruano, como Porras Barrenechea, Pulgar Vidal, Mejía Xesspe y Linares Málaga, quienes han seguido ciertamente claros parámetros científicos en el tratamiento y análisis de estos preciado bienes culturales; éste es el espíritu que debe poblar las mentes de los que quieren proteger el legado nacional de nuestra historia, y de los que quieren entender el desarrollo cognitivo de los antiguos peruanos plasmados en sus piedras; este es el espíritu de los que quieren leer nuevamente las guacas del Perú. REFERENCIAS BIBLIOGRÁFICAS Alvarez, A. M., (2006). Arte rupestre paleolítico (I): soportes, técnicas y temática. En: Prehistoria y Protohistoria de la Península Ibérica, pp 378-405. Tomo I. Unidad Didáctica. Universidad Nacional de Educación a Distancia. Madrid. Asociación Peruana de Arte Rupestre. (2007), Código de Ética Para Visitas a Sitios Arqueológicos con Arte Rupestre. En: httn:/ /grouns.google.com /mono/spar neru/web/codigode-etica Bednarik, R.. (1998), The Technology of the petroglyphs. Rock Art Research (15) 1: 23-35. Bedn ik, R., (2007), Rock Art Science, The Scientific Study of Palaeoart. Aryan Book International, New Delhi. Bueno, A.. (2006), Petroglifos en la quebrada Marín y La Galgada: de los textos gráficos al mito etiológico. Investigaciones Sociales. X(17): 67-90. 155N 2312-9115; 20140) Vol. ¡11 :3847 Campana, C., (2007), El prisionero del tiempo: un petroglifo del Ato de las Guitarras". En: Hostnig, Strecker y Guffroy (Mg.), Actas del Primer Simposio Nacional de Arte Rupestre del Perú (Cuco noviembre 2004), pp. 37-67. Antes y Memoires 12. IFEA. Lima. Cardich, A., (1964), Lauricocha. Fundamentos para una prehistoria de los andes centrales. En: Studia Prehistórica III. Centro Argentino de Estudios Prehistóricos. Buenos Aires. 171 pp. Echevarría, G., (2003), Petroglifos en la cuenca del río Cachiyacu, metodología y registro en contexto de explotación petrolera. VI Simposio Internacional de Arte Rupestre, Jujuy, 29 noviembre al 4 de diciembre. Argentina. Echevarría, G., (2004), El petrograbado de Chocas, costa central del Perú". ter Simposio Nacional de Arte Rupestre. Cosco, 25 al 30 de noviembre. Perú. Echevarría, G., (2008), Rock art in Peru, problems and perspectives. Man in India: A quarterly International Journal of Anthropology 88(2-3): 261-275. Hostnig, R., (2003), Arte Rupestre del Perú. Inventario Nacional. CONCYTEC. Lima. Hutchinson, T. J., (1873), Two Years in Peru, with Exploration of its Antiquities. Vol. II. London. Linares, E., (1960), Notas sobre los petroglifos de Toro Muerto. En: Antiguo Perú, Espacio y Tiempo, pp. 297-299. Juan Mejía Baca editor. Lima. Linares, E., (1973), Anotaciones sobre las cuatro modalidades de arte rupestre en Arequipa (pictografías, Petroglifos, Arte rupestre moblar y Geoglifos). Anales Científicos de la Universidad del Centro del Perú. 2:133-267. Linares, E., (1974), Arte Rupestre: El Arte Rupestre en el Sur del Perú. Introducción al Estudio de los Petroglifos de Toro Muerto. Tesis Doctoral, 2 tomos. 700 pp., 223 ilustraciones, 2 mapas. Universidad San Agustín. Arequipa. Huancayo. Linares, E., (1988), Arte mobiliar con tradición rupestre en el sur del Perú. Rock Art Research 5: 54-65. Linares, E.. (1999), Arte Rupestre en Sudamérica Prehistoria. Universidad Nacional Mayor de San Marcos. Fondo Editorial. Lima, Perú. Mejía, T., (1968), Pintura Chavinoide en los lindes del arte rupestre. San Marcos 19:15-32. Lima. Morales, D., (1993), Historia Arqueológica del Perú. En: Compendio Histórico del Perú, pp. 138-149. Tomo I. Editorial Milla Batres. Lima. Muelle, J., (1969), Las cuevas y pinturas de Toquepala. En: Mesa Redonda de Ciencias Prehistóricas y Antropológicas. PUCPIRA. Tomo II. pp. 186-196. Lima. Neyra, M., (1968), Un complejo lítico y pinturas rupestres en la gruta Su-3 de Sumbay. Revista de la Facultad de Letras. 5: 43-75. Arequipa. Nieves, A., (2006), Reconstructing ritual: some thoughts on the location of petroglyphs groups in the Nasca valley, Pera. In: Robertson, Elizabeth, et al. In Space and Spatial Analysis in Archae,ology, pp. 217-226. Calgary Press. Canada. Nuñez, A., (1986), Petroglifos del Perú. Panorama Mundial del Arte Rupestre. Editorial Científico Técnica. La Habana. Porras, R., (1963), Fuentes Históricas Peruanas. Instituto Raúl La Vida y la Nidada Echevarría, G. Las cuatro categorías técnico materiales del Arte Rupestre Peruano Porras Barrenechea, Universidad Nacional Mayor de San Marcos. Lima. Porras, R., (1999), Indagaciones Peruanas, El Legado Quechua. Universidad Nacional Mayor de San Marcos, Fondo Editorial. Lima. Querejazu, R., (2001), El Arte Rupestre de la Cuenca del Río Mizgue. Universidad Mayor de San Simón, Facultad de Arquitectura, Instituto de Investigaciones. Cochabamba. Pulgar, J., (1946), Historia y Geografía del Perú Tomo L Las Ocho Regiones Naturales del Perú. Universidad Nacional Mayor de San Marcos, Lima. Pulgar, 1, (1959/1960), Introducción. La investigación Toponímica y el hallazgo de los centros pictográficos de la cuenca del río Huallaga. Revista del Instituto de Geografía 6: 155-156. Universidad Nacional Mayor de San Marcos, Facultad de Letras. Lima, Perú. llavines, R., (1986) Arte Rupestre en el Perú, Inventario General. (Primera Aproximación). Instituto Nacional de Cultura. Lima. Rivero, M. de y De Thchudi, J., (1851) [1958]. Antigüedades Peruanas. Primer Festival del Libro Arequipeño. Arequipa. Shady, R., (2000), Los orígenes de la civilización y la formación del estado en el Perú. Boletín del Museo de Arqueología y Antropología 3(2): 2-7. Tallo, J. 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https://openalex.org/W2899839204	OpenAlex	Open Science	CC-By	2,018	Measurement of jet fragmentation in Pb+Pb and <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:mi>p</mml:mi><mml:mi>p</mml:mi></mml:mrow></mml:math> collisions at <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:msqrt><mml:msub><mml:mi>s</mml:mi><mml:mrow><mml:mi>N</mml:mi><mml:mi>N</mml:mi></mml:mrow></mml:msub></mml:msqrt><mml:mo>=</mml:mo><mml:mn>5.02</mml:mn></mml:mrow></mml:math> TeV with the ATLAS detector	Morad Aaboud	English	Spoken	30,720	67,383	Measurement of jet fragmentation in Pb plus Pb and pp collisions at sNN = 5.02 TeV with th ATLAS detector Doi link to publisher: https://doi.org/10.1103/PhysRevC.98.024908 Version of the following full text: Publisher’s version Downloaded from: http://hdl.handle.net/2066/195388 Download date: 2024-10-23 Measurement of jet fragmentation in Pb plus Pb and pp collisions at sNN = 5.02 TeV with the ATLAS detector Aaboud, M.; Aad, G.; Abbott, B.; Abdinov, O.; Abeloos, B.; Abhayasinghe, D.K.; Alderweireldt, S.C.; Caron, S.; Colasurdo, L.; Groot, N. de; Fabiani, V.; Filthaut, F.; Galea, C.; Igonkina, O.; König, A.C; Nektarijevic, S.; Pedraza Diaz, L.; Schouwenberg, J.F.P.; Zwalinski, L. 2018, Article / Letter to editor (Physical Review C, 98, 2, (2018), pp. 1-34, article 024908) Doi link to publisher: https://doi.org/10.1103/PhysRevC.98.024908 Measurement of jet fragmentation in Pb plus Pb and pp collisions at sNN = 5.02 TeV with the ATLAS detector Aaboud, M.; Aad, G.; Abbott, B.; Abdinov, O.; Abeloos, B.; Abhayasinghe, D.K.; Alderweireldt, S.C.; Caron, S.; Colasurdo, L.; Groot, N. de; Fabiani, V.; Filthaut, F.; Galea, C.; Igonkina, O.; König, A.C; Nektarijevic, S.; Pedraza Diaz, L.; Schouwenberg, J.F.P.; Zwalinski, L. 2018, Article / Letter to editor (Physical Review C, 98, 2, (2018), pp. 1-34, article 024908) Doi link to publisher: https://doi.org/10.1103/PhysRevC.98.024908 I. INTRODUCTION The distribution of particles within the jet are affected by this mechanism of energy loss. Several related observables sensitive to the properties of the medium can be constructed. Measurements of the jet shape [13] and the fragmentation functions were made in 2.76 TeV Pb+Pb collisions [14–16]. In Ref. [16], jet fragmentation functions are measured as a function of both the charged-particle transverse momentum pT and the charged-particle longitudinal momentum fraction relative to the jet, Ultrarelativistic nuclear collisions at the Large Hadron Col- lider (LHC) produce hot dense matter called the quark-gluon plasma (QGP); recent reviews can be found in Refs. [1,2]. Hard-scattering processes occurring in these collisions pro- ducejetswhichtraverseandinteractwiththeQGP.Thestudyof modiﬁcations of jet rates and properties in heavy-ion collisions compared to pp collisions provides information about the properties of the QGP. z ≡pT cos R / pjet T . (1) (1) The rates of jet production are observed to be reduced by approximately a factor of 2 in lead-lead (Pb+Pb) colli- sions at LHC energies compared to expectations from the jet production cross sections measured in pp interactions scaled by the nuclear overlap function of Pb+Pb collisions [3–5]. Similarly, back-to-back dijet [6–8] and photon-jet pairs [9] are observed to have unbalanced transverse momentum in Pb+Pb collisions compared to pp collisions. Related phenomena were ﬁrst observed at the Relativistic Heavy Ion Collider where the measurements were made with hadrons rather than reconstructedjets[10–12]. Theseobservationsimplythatsome of the energy of the parton showering process is transferred outside of the jet through its interaction with the QGP. This has been termed “jet quenching.” The fragmentation functions are deﬁned as The fragmentation functions are deﬁned as D(z) ≡ 1 Njet dnch dz , and D(pT) ≡ 1 Njet dnch dpT , where pjet T is the transverse momentum of the jet, nch is the number of charged particles in the jet, Njet is the number of jets under consideration, and R = (η)2 + (φ)2 with η and φ deﬁned as the differences between the jet axis and the charged-particle direction in pseudorapidity and azimuth,1 1ATLAS uses a right-handed coordinate system with its origin at the nominal interaction point (IP) in the center of the detector and the z axis along the beam pipe. The x axis points from the IP to the center of the LHC ring, and the y axis points upward. (Received 16 May 2018; published 16 August 2018) This paper presents a measurement of jet fragmentation functions in 0.49 nb−1 of Pb+Pb collisions and 25 pb−1 of pp collisions at √sNN = 5.02 TeV collected in 2015 with the ATLAS detector at the LHC. These measurements provide insight into the jet quenching process in the quark-gluon plasma created in the aftermath of ultrarelativistic collisions between two nuclei. The modiﬁcations to the jet fragmentation functions are quantiﬁed by dividing the measurements in Pb+Pb collisions by baseline measurements in pp collisions. This ratio is studied as a function of the transverse momentum of the jet, the jet rapidity, and the centrality of the collision. In both collision systems, the jet fragmentation functions are measured for jets with transverse momentum between 126 and 398 GeV and with an absolute value of jet rapidity less than 2.1. An enhancement of particles carrying a small fraction of the jet momentum is observed, which increases with centrality and with increasing jet transverse momentum. Yields of particles carrying a very large fraction of the jet momentum are also observed to be enhanced. Between these two enhancements of the fragmentation functions a suppression of particles carrying an intermediate fraction of the jet momentum is observed in Pb+Pb collisions. A small dependence of the modiﬁcations on jet rapidity is observed. Note: To cite this publication please use the final published version (if applicable). PHYSICAL REVIEW C 98, 024908 (2018) Measurement of jet fragmentation in Pb+Pb and pp collisions at √sN N = 5.02 TeV with the ATLAS detector M. Aaboud et al.∗ (ATLAS Collaboration) ©2018 CERN, for the ATLAS Collaboration I. INTRODUCTION Cylindrical coordinates (r, φ) are used in the transverse plane, φ being the azimuthal angle Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. 024908-1 ©2018 CERN, for the ATLAS Collaboration 2469-9985/2018/98(2)/024908(34) PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. respectively. In order to quantify differences between Pb+Pb and pp collisions at the same collision energy, the ratios of the fragmentation functions are measured: respectively. In order to quantify differences between Pb+Pb and pp collisions at the same collision energy, the ratios of the fragmentation functions are measured: rapidityrangeof\|yjet\| < 2.1usingtheanti-kt reconstructional- gorithm[24] withradius parameterR = 0.4. Themeasurement is presented in intervals of pjet T , yjet, and collision centrality. These data extend the previous studies at √sNN = 2.76 TeV in two ways. First, an increase in the peak energy density of the medium is expected. Second, the Pb+Pb integrated luminosity in the current dataset is 3.5 times the integrated luminosity available at 2.76 TeV, and the increase in the collision energy also increases the jet cross sections. These two factors allow a measurement of the dependence of jet fragmentation functions on the transverse momentum of the jet over a wider range than was previously possible. RD(z) ≡D(z)PbPb D(z)pp , and and and RD(pT) ≡D(pT)PbPb D(pT)pp . Relative to jets in pp collisions, it was found in Ref. [16] that jets in Pb+Pb collisions have an excess of particles with transverse momentum below 4 GeV and an excess of particles carrying a large fraction of the jet transverse momentum. At intermediate charged-particle pT, there is a suppression of the charged-particle yield. At the same time, an excess of low-pT particles is observed for particles in a wide region around the jet cone [17,18]. These observations may indicate that the energy lost by jets through the jet quenching process is being transferred to soft particles within and around the jet [19,20]; measurements of these soft particles have the potential to constrain the models describing such processes. A possible explanation for the enhancement of particles carrying a large fraction of the jet momentum is that it is related to the gluon-initiated jets losing more energy than quark- initiated jets. II. EXPERIMENTAL SETUP The measurements presented in this paper were performed using the ATLAS inner detector, calorimeter, trigger, and data acquisition systems [25]. The calorimeter system consists of a sampling liquid argon (LAr) electromagnetic (EM) calorime- ter covering \|η\| < 3.2, a steel/scintillator sampling hadronic calorimeter covering \|η\| < 1.7, LAr hadronic calorimeters covering 1.5 < \|η\| < 3.2, and two LAr forward calorimeters (FCal) covering 3.1 < \|η\| < 4.9 [25]. The EM calorimeters are segmented longitudinally in shower depth into three layers with an additional presampler layer. They have segmentation in φ and η that varies with layer and pseudorapidity. The hadronic calorimeters have three sampling layers longitudinal in shower depth. The inner detector measures charged particles within the pseudorapidity interval \|η\| < 2.5 using a combination of sil- icon pixel detectors, silicon microstrip detectors (SCTs), and a straw-tube transition radiation tracker (TRT), all immersed in a 2 T axial magnetic ﬁeld [25]. Each of the three de- tectors is composed of a barrel and two symmetric endcap sections. The pixel detector is composed of four layers: the “insertable B layer” [26,27] and three layers with a pixel size of 50 μm × 400 μm. The SCT barrel section contains four layers of modules with 80 μm pitch sensors on both sides and each endcap consists of nine layers of double-sided modules with radial strips having a mean pitch of 80 μm. The two sides of eachSCTlayerinboththebarrelandtheendcapshavearelative stereo angle of 40 mrad. The TRT contains up to 73 (160) layers of staggered straws interleaved with ﬁbers in the barrel (endcap). Proton-nucleus collisions, which do not generate a large amount of QGP, are used to differentiate between initial- and ﬁnal-state effects due to the QGP formed in Pb+Pb collisions. Fragmentation functions in p+Pb collisions show no evidence of modiﬁcation when compared with those in pp collisions [22]. Thus, any modiﬁcations observed in Pb+Pb collisions can be attributed to the presence of the QGP rather than to effects arising from the presence of the large nucleus. The rapidity dependence of jet observables in Pb+Pb collisions is of great interest, in part because at ﬁxed pjet T the fraction of quark jets increases with increasing \|yjet\| (see, for example,Refs.[21,23]).Thismakestherapiditydependenceof jet observables potentially sensitive to the different interactions of quarks and gluons with the QGP. I. INTRODUCTION This leads to a higher quark-jet fraction in Pb+Pb collisions than in pp collisions. The change in ﬂavor compo- sition combined with the different shapes of the quark and gluon fragmentation functions [21] then lead to the observed excess. II. EXPERIMENTAL SETUP Previous measurements of the rapidity dependence of jet fragmentation functions at √sNN = 2.76 TeV in Pb+Pb collisions found a rapidity dependence of the fragmentation function modiﬁcation with limited signiﬁcance [16]. The zero-degree calorimeters (ZDCs) are located sym- metrically at z = ±140 m and cover \|η\| > 8.3. They are constructed from tungsten absorber plates and ˇCerenkov light is transmitted via quartz ﬁbers. In Pb+Pb collisions the ZDCs primarily measure “spectator” neutrons, i.e., neutrons that do not interact hadronically when the incident nuclei collide. A ZDC coincidence trigger is implemented by requiring the pulse height from each ZDC to be above a threshold set to accept the single-neutron peak. In this paper, the fragmentation functions and the RD(z) and RD(pT) ratios are measured in Pb+Pb and pp collisions at 5.02 TeV using 0.49 nb−1 of Pb+Pb collisions and 25 pb−1 of pp collisions collected in 2015. Jets are measured over a A two-level trigger system is used to select the Pb+Pb and pp collisions. The ﬁrst trigger level (L1) is hardware-based and implemented with custom electronics. The second level is the software-based high-level trigger (HLT) and is used to further reduce the accepted event rate. Minimum-bias Pb+Pb events are recorded using a trigger deﬁned by the logical OR around the beam pipe. The pseudorapidity is deﬁned in terms of the polar angle θ as η = −ln tan(θ/2). The rapidity is deﬁned as y = 0.5 ln[(E + pz)/(E −pz)] where E and pz are the energy and the component of the momentum along the beam direction. 024908-2 PHYSICAL REVIEW C 98, 024908 (2018) MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … NNPDF23LO parton distribution function (PDF) set [32]. The detector response was simulated using GEANT4 [33,34] and the simulated hits were combined with those from the data event. A weight is assigned to each MC event such that the event sample obtained from the minimum-bias trigger has the same centrality distribution as the sample collected by the jet trigger. A separate sample of 1.8 × 107 simulated 5.02 TeV PYTHIA8 pp hard-scattering events, generated with the same tune and PDFs as for the Pb+Pb MC sample, is used to evaluate the performance for measuring fragmentation functions in the pp data. The contribution from additional collisions in the same bunch crossing is not included in the MC simulation. III. DATA SETS AND EVENT SELECTION The Pb+Pb and pp data used in this analysis were recorded in 2015. The data samples consist of 25 pb−1 of √s = 5.02 TeV pp data and 0.49 nb−1 of √sNN = 5.02 TeV Pb+Pb data. In Pb+Pb and pp collisions, events are required to have a reconstructed vertex within 150 mm of the nominal interaction point along the beam axis. Only events taken during stable beam conditions and satisfying detector and data-quality re- quirements, which include the calorimeters and inner tracking detectors being in nominal operation, are considered. In Pb+Pb collisions, the event centrality reﬂects the overlap area of the two colliding nuclei and is characterized by EFCal T , the total transverse energy deposited in the FCal [28]. The centrality intervals used in this analysis are deﬁned according to successive percentiles of the EFCal T distribution obtained from minimum-bias triggered Pb+Pb events ordered from the most central (highest EFCal T ) to the most peripheral collisions (lowest EFCal T ): 0–10%, 10–20%, 20–30%, 30–40%, 40– 60%, 60–80%. In addition to the jet-triggered sample, a separate Pb+Pb data sample was recorded with the minimum-bias trigger and two total transverse-energy triggers requiring 1.5 and 6.5 TeV to enhance the rate of more central Pb+Pb events. This data sample is used to produce a Pb+Pb Monte Carlo (MC) events with conditions that match those registered while the data were recorded. Jets are required to have a rapidity within \|yjet\| < 2.1 so that all R = 0.4 jet cones are contained within the inner detector’s acceptance. To prevent neighboring jets from distorting the measurement of the fragmentation functions, jets are rejected if there is another jet with higher pjet T anywhere within a distance R < 1.0. A correction is applied to reduce the effects of the broadening of the jet direction measurement for R = 0.4 jets due to the UE. The correction uses jets reconstructed with a smaller distance parameter R = 0.2 since their angular resolution evaluated in MC studies is found to be less affected by the UE ﬂuctuations than that of larger-R jets. The jet direction is redeﬁned as that of the closest R = 0.2 jet with The performance of the detector and of the analysis procedure in Pb+Pb collisions is evaluated using 1.8 × 107 5.02 TeV MC events. II. EXPERIMENTAL SETUP A sample of Pb+Pb events generated with HIJING version 1.38b [35] is also used to evaluate the performance of the track reconstruction. of a L1 total energy trigger and the ZDC coincidence trigger. The total energy trigger required the total transverse energy measured in the calorimeter system to be greater than 50 GeV in Pb+Pb collisions. Jet events are selected by the HLT, after requiring the identiﬁcation of a jet by the L1 jet trigger in pp collisions or the total energy trigger with a threshold of 50 GeV in Pb+Pb collisions. The L1 jet trigger utilized in pp collisions required a jet with transverse momentum greater than 20 GeV. The HLT jet trigger used a jet reconstruction algorithm similar to that used in the ofﬂine analysis (the ofﬂine jet reconstruction is discussed in Sec. IV). It selected events containing jets with transverse energy of at least 75 GeV in Pb+Pb collisions and at least 85 GeV in pp collisions. In pp collisions, the 85 GeV threshold jet trigger sampled the full delivered luminosity. The 75 GeV threshold jet trigger used in Pb+Pb collisions was prescaled2 in a small part of the Pb+Pb data-taking period; however, the trigger sampled more than 99% of the total integrated luminosity. The measurement is performed in the jet transverse momentum region where the triggers are fully efﬁcient. 2The prescale indicates which fraction of events that passed the trigger selection was selected for recording by the data acquisition system. IV. JET AND TRACK SELECTION The jet reconstruction, underlying event (UE) determina- tion, and subtraction procedures closely follow those used by ATLAS for jet measurements in pp and Pb+Pb collisions at √sNN = 2.76 TeV [4]. The anti-kt algorithm is ﬁrst run in four- momentum recombination mode, on η × φ = 0.1 × 0.1 calorimeter towers with the anti-kt radius parameter R = 0.2 and R = 0.4. The energies in the towers are obtained by sum- ming the energies of calorimeter cells at the electromagnetic energy scale within the tower boundaries. Then, an iterative procedure is used to estimate the η-dependent UE transverse energy density on an event-by-event basis using the energy measurements in all calorimeter towers in the event while excluding the regions populated by jets. The resulting UE transverse energy density is modulated taking into account the presence of the azimuthal anisotropy of particle production [36]. The modulation includes contributions of the second-, third-, and fourth-order azimuthal anisotropy harmonics. Higher-order harmonics introduce negligible variation of the reconstructed jet energy. The UE transverse energy is sub- tracted from each calorimeter cell within the towers included in the reconstructed jet, and the four-momentum of the jet is updated accordingly. Then, a jet η- and pT-dependent correc- tion factor to the pjet T derived from the simulation samples is applied to correct for the calorimeter energy response [37]. An additional correction based on in situ studies of jets recoiling against photons, Z bosons, and jets in other regions of the calorimeter is applied [38,39]. The same jet reconstruction procedure without the azimuthal modulation of the UE is also applied to pp collisions. j III. DATA SETS AND EVENT SELECTION The efﬁciency exhibits only a small variation with yjet in the region \|yjet\| < 1.2, and it decreases by approximately 10% in the most forward yjet interval. pjet T > 35 GeV and matching the original jet direction within R = 0.3 of the R = 0.4 jet, when such a matching jet is found. If no matching R = 0.2 jet is found the axis remains unchanged. g Charged-particle tracks are reconstructed from hits in the inner detector using the track reconstruction algorithm with settings optimized for the high hit density in heavy-ion colli- sions [40]. Tracks used in this analysis are required to have a total of at least 9 (11) hits in the silicon pixel and microstrip detectors for charged particles with pseudorapidity \|ηch\| ⩽ 1.65(\|ηch\| > 1.65). At least one hit is required in one of the two innermost pixel layers. If the track trajectory passed through an active module in the innermost layer, then a hit in this layer is required. Furthermore, a track must have no more than two holes in the Pixel and SCT detectors together, where a hole is deﬁned by the absence of a hit predicted by the track trajectory. All charged-particle tracks used in this analysis are required to have reconstructed transverse momentum pch T > 1 GeV. In order to suppress the contribution from secondary particles, the distance of closest approach of the track to the primary vertex in the transverse plane is required to be less than a value which varies from 0.45 mm at pch T = 4 GeV to 0.2 mm at pch T = 20 GeV, and at that point the track must be less than 1.0 mm from the primary vertex in the longitudinal direction. The contribution of reconstructed tracks which are not be matched to a generated primary particle in the MC samples of pp collision events produced without data overlay, along with the residual contribution of tracks matched to secondary particles, are together considered “fake” tracks. The fraction of fake tracks is less than 2% over the full kinematic range of this measurement. A possible degradation of the tracking performance at high occupancy is checked in the sample of Pb+Pb collision events simulated with the HIJING MC. No signiﬁcant dependence of the rate of fake tracks on centrality is observed. The correction for the fake contribution is discussed in Sec. V. III. DATA SETS AND EVENT SELECTION [GeV] truth T p 1 10 2 10 Tracking efficiency 0.7 0.75 0.8 0.85 0.9 0.95 1 ATLAS Simulation = 5.02 TeV s pp < 158 GeV jet T p 126 GeV < < 200 GeV jet T p 158 GeV < < 251 GeV jet T p 200 GeV < < 316 GeV jet T p 251 GeV < < 398 GeV jet T p 316 GeV < \| < 0.3 jet y \| =0.4 R tk anti- [GeV] truth T p 1 10 2 10 Tracking efficiency 0.7 0.75 0.8 0.85 0.9 0.95 1 < 158 GeV jet T p 0-10%, 126 GeV < < 316 GeV jet T p 0-10%, 251 GeV < < 158 GeV jet T p 60-80%, 126 GeV < < 316 GeV jet T p 60-80%, 251 GeV < ATLAS Simulation = 5.02 TeV NN s Pb+Pb \| < 0.3 jet y \| =0.4 R tk anti- FIG. 1. Tracking efﬁciency ε, smoothed using a third-order polynomial in ln(ptruth T ) as a function of ptruth T in pp collisions in ﬁve different jet-pT intervals (left) and in Pb+Pb collisions (right) in two different jet-pT intervals and for 0–10% and 60–80% centrality intervals. In both plots the efﬁciency is evaluated for tracks within jets with \|yjet\| < 0.3. reconstruction efﬁciencies as a function of the generator-level primary particle transverse momentum, ptruth T , are shown in Fig. 1 for jets with \|yjet\| < 0.3 in pp and Pb+Pb collisions. In order to remove ﬂuctuations in the efﬁciency due to the limited MC sample size, the ptruth T dependence of the efﬁciencies is parametrized and smoothed using a third-order polynomial in ln(ptruth T ) that gives a good description of the efﬁciency in the full range of ptruth T . The efﬁciencies shown in Fig. 1 exhibit only a modest variation with ptruth T , centrality, and pjet T . A small almost continuous increaseof theefﬁciencywiththeincreasing ptruth T is observed. The efﬁciency over the 20–100 GeV ptruth T range is smaller for high pjet T compared to low pjet T by about 2% and 5% in pp and Pb+Pb collisions, respectively. This behavior is attributed to the higher probability to lose tracks in the dense core of high-pT jets than to lose tracks that are more isolated [41]. The efﬁciency is lower in more central Pb+Pb collisions due to the higher hit density. III. DATA SETS AND EVENT SELECTION The efﬁciency, ε(ptruth T , pjet T , yjet), for reconstructing charged particles within jets in Pb+Pb and pp collisions is evaluated from the matching of reconstructed tracks to generator-levelprimaryparticles3 usingMCsamplesdescribed above. The matching is based on contributions of generator- level particles to the hits in the detector layers. A reconstructed track is matched to a generator-level particle if it contains hits produced primarily by this particle [34]. The efﬁciency is evaluated separately in four \|yjet\| intervals and each interval of reconstructed pjet T used in the measurement. Furthermore, the efﬁciency is evaluated separately for each centrality in- terval in the case of Pb+Pb collisions. The charged-particle III. DATA SETS AND EVENT SELECTION These were produced from minimum- bias Pb+Pb data events overlaid with hard-scattering dijet pp events generated with POWHEG+PYTHIA8 [29,30] using a set of tuned parameters called the A14 tune [31] and the 2The prescale indicates which fraction of events that passed the trigger selection was selected for recording by the data acquisition system. 024908-3 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. [GeV] truth T p 1 10 2 10 Tracking efficiency 0.7 0.75 0.8 0.85 0.9 0.95 1 ATLAS Simulation = 5.02 TeV s pp < 158 GeV jet T p 126 GeV < < 200 GeV jet T p 158 GeV < < 251 GeV jet T p 200 GeV < < 316 GeV jet T p 251 GeV < < 398 GeV jet T p 316 GeV < \| < 0.3 jet y \| =0.4 R tk anti- [GeV] truth T p 1 10 2 10 Tracking efficiency 0.7 0.75 0.8 0.85 0.9 0.95 1 < 158 GeV jet T p 0-10%, 126 GeV < < 316 GeV jet T p 0-10%, 251 GeV < < 158 GeV jet T p 60-80%, 126 GeV < < 316 GeV jet T p 60-80%, 251 GeV < ATLAS Simulation = 5.02 TeV NN s Pb+Pb \| < 0.3 jet y \| =0.4 R tk anti- FIG. 1. Tracking efﬁciency ε, smoothed using a third-order polynomial in ln(ptruth T ) as a function of ptruth T in pp collisions in ﬁve different jet-pT intervals (left) and in Pb+Pb collisions (right) in two different jet-pT intervals and for 0–10% and 60–80% centrality intervals. In both plots the efﬁciency is evaluated for tracks within jets with \|yjet\| < 0.3. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … [GeV] T p 1 10 ) T p ( D / sub ) ch T p ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- [GeV] T p 1 10 2 10 ) T p ( D / sub ) ch T p ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 316 GeV jet T p 251 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- [GeV] T p 1 10 ) T p ( D / sub ) ch T p ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- z 2 − 10 1 − 10 1 ) z( D / sub ) z ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- z 2 − 10 1 − 10 1 ) z( D / sub ) z ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- [GeV] T p 1 10 ) T p ( D / sub ) ch T p ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- z 2 − 10 1 − 10 1 ) z ( D / sub ) z ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 316 GeV jet T p 251 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- [GeV] T p 1 10 2 10 ) T p ( D / sub ) ch T p ( D 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 , 0-10% -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 316 GeV jet T p 251 < ATLAS \|<2.1 jet y =0.4 jets, \| R tk anti- FIG. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) FIG. 2. Ratio of the measured charged-particle distributions before and after the subtraction of the UE and fake tracks as a function of pch T for pjet T in the range 126–158 GeV for 0–10% (left), 30–40% (middle), and 60–80% (right) centrality. The uncertainties are smaller than the marker size in all cases for which there is a signiﬁcant UE. FIG. 2. Ratio of the measured charged-particle distributions before and after the subtraction of the UE and fake tracks as a function of pch T for pjet T in the range 126–158 GeV for 0–10% (left), 30–40% (middle), and 60–80% (right) centrality. The uncertainties are smaller than the marker size in all cases for which there is a signiﬁcant UE. FIG. 2. Ratio of the measured charged-particle distributions before and after the subtraction of the UE and fake tracks as a function of pch T for pjet T in the range 126–158 GeV for 0–10% (left), 30–40% (middle), and 60–80% (right) centrality. The uncertainties are smaller than the marker size in all cases for which there is a signiﬁcant UE. constructed as Tracks which are not correlated with the jet need to be subtracted from the measured distributions; these tracks come from both fake tracks and the UE. In Pb+Pb collisions, contributions to the fragmentation functions from the charged particles originating from the UE in Pb+Pb collisions are subtracted. This contribution is evaluated as a function of charge particle z or pch T , yjet, pjet T , and the collision centrality. Additionally, the measured track yields in pp and Pb+Pb collisions are corrected for the presence of fake tracks. ucted as dnmeas ch dz = Nch z, yjet, pjet T z dnmeas ch dpch T = Nch pch T , yjet, pjet T pch T , and dnmeas ch dpch T = Nch pch T , yjet, pjet T pch T , where the quantities Nch(z) and Nch(pch T ) represent the number of associated tracks within the given z or pch T range, respectively corrected for the track reconstruction efﬁciency. The efﬁciency correction is applied as a 1/ε(pch T , pjet T , yjet) weight on a track-by-track basis, assuming pch T = ptruth T . MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … While that assumption is not strictly valid, the efﬁciency varies sufﬁciently slowly with ptruth T that the error introduced by this assumption is less than 1%. where the quantities Nch(z) and Nch(pch T ) represent the number of associated tracks within the given z or pch T range, respectively corrected for the track reconstruction efﬁciency. The efﬁciency correction is applied as a 1/ε(pch T , pjet T , yjet) weight on a track-by-track basis, assuming pch T = ptruth T . While that assumption is not strictly valid, the efﬁciency varies sufﬁciently slowly with ptruth T that the error introduced by this assumption is less than 1%. The UE contribution is determined for each measured jet using a grid of R = 0.4 cones spanning the full coverage of the inner detector and following the method introduced in Ref. [14]. The method is applied to events containing jets included in the analysis. The cones have a ﬁxed distance between their centers chosen such that the inner detector acceptance is uniformly covered while avoiding overlaps. V. ANALYSIS PROCEDURE The analysis procedure closely follows the one used in the measurement of jet fragmentation at √sNN = 2.76 TeV [16]. Reconstructed tracks are associated with a reconstructed jet if they fall within R = 0.4 of the jet axis and for each of these particles the longitudinal momentum fraction z is calculated. The measured track yields, dnmeas ch /dz or dnmeas ch /dpch T , are 3Primary particles are deﬁned as particles with a mean lifetime τ > 0.3 × 10−10 s either directly produced in pp interactions or from subsequent decays of particles with a shorter lifetime. All other particles are considered to be secondary. 024908-4 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … 3. Ratios Dsub(z)/D(z) (left) and Dsub(pch T )/D(pT) (right) for pp and 0–10% central Pb+Pb collisions for 126 < pjet T < 158 GeV (top) and 251 < pjet T < 316 GeV (bottom) for \|yjet\| < 2.1. The error bars show the statistical uncertainties and the boxes show the systematic uncertainties in the unfolding procedure. 1 z FIG. 3. Ratios Dsub(z)/D(z) (left) and Dsub(pch T )/D(pT) (right) for pp and 0–10% central Pb+Pb collisions for 126 < pjet T < 158 GeV (top) and 251 < pjet T < 316 GeV (bottom) for \|yjet\| < 2.1. The error bars show the statistical uncertainties and the boxes show the systematic uncertainties in the unfolding procedure. 024908-5 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. z 2 − 10 1 − 10 1 [%] D(z) δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS = 5.02 TeV NN s -1 Pb+Pb 2015, 0.49 nb < 158 GeV jet T p 126 < 0-10% z 2 − 10 1 − 10 1 [%] D(z) δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS = 5.02 TeV s -1 2015, 25 pb pp < 158 GeV jet T p 126 < [GeV] T p 1 10 ) [%] T p ( D δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS = 5.02 TeV NN s -1 Pb+Pb 2015, 0.49 nb < 158 GeV jet T p 126 < 0-10% [GeV] T p 1 10 ) [%] T p ( D δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS = 5.02 TeV s -1 2015, 25 pb pp < 158 GeV jet T p 126 < FIG. 4. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Summary of the systematic uncertainties of the D(z) (top) and D(pT) (bottom) distributions in 0–10% central Pb+Pb collisions (left) and pp collisions (right) for jets in the 126–158 GeV pjet T interval. The systematic uncertainties due to JES, JER, unfolding, UE contribution, MC nonclosure and tracking are shown along with the total systematic uncertainty from all sources. z 2 − 10 1 − 10 1 [%] D(z) δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS = 5.02 TeV NN s -1 Pb+Pb 2015, 0.49 nb < 158 GeV jet T p 126 < 0-10% z 1 [GeV] T p FIG. 4. Summary of the systematic uncertainties of the D(z) (top) and D(pT) (bottom) distributions in 0–10% central Pb+Pb collisions (left) and pp collisions (right) for jets in the 126–158 GeV pjet T interval. The systematic uncertainties due to JES, JER, unfolding, UE contribution, MC nonclosure and tracking are shown along with the total systematic uncertainty from all sources. Any cone having a charged particle with pch T > 10 GeV or overlapping with a reconstructed jet with pjet T > 90 GeV is assumed to be associated with a hard process and is excluded from the UE estimation to avoid biasing it. The parameters deﬁning the exclusion regions are evaluated in MC studies and are subjected to variations as part of the estimation of systematic uncertainties. The resulting UE charged particle yields, dnUE ch /dz or dnUE ch /dpch T , are evaluated over 1 < pch T < 10 GeV according to applied to the charged-particle UE estimate to account for the difference in the azimuthal particle density, due to elliptic ﬂow, between the φ angle of the cone and the φ angle of the jet. This utilizes a centrality- and pch T -dependent parametrization of the measured elliptic ﬂow coefﬁcients [36]. The UE contribution is further corrected for the correlation between the actual UE charged particle yield underneath the jet and the jet energy resolution [14]; in regions where the UE has an upward ﬂuctuation, the jet energy resolution is worse. The smearing due to jet energy resolution leads to a net migration of jets from lower pjet T to higher pjet T values. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … The effect of the migration causes the actual UE contribution underneath the jet to be larger than that estimated from the procedure described above. This effect is corrected for by applying multiplicative correction factors, depending on pch T or z, yjet, pjet T , and collision centrality. The correction is estimated as a ratio of the UE charged particle yield evaluated by two different methods using the Pb+Pb MC samples. The ﬁrst estimate uses the cone method discussed above. The second method calculates the UE contribution in the data overlay MC samples from tracks, within the area of a jet, that do not have an associated generated primary particle. The size of the correction is less than 2% at low z or pch T where the UE has the largest impact, and has only a small dependence on pjet T . dnUE ch dz = 1 Ncone 1 ε pch T ,ηch Ncone ch z,pjet T ,yjet z z=pch T cos R/pjet T , dnUE ch dpch T = 1 Ncone 1 ε pch T , ηch Ncone ch pch T , pjet T , yjet pch T . dnUE ch dz = 1 Ncone 1 ε pch T ,ηch Ncone ch z,pjet T ,yjet z z=pch T cos R/pjet T , dnUE ch dpch T = 1 Ncone 1 ε pch T , ηch Ncone ch pch T , pjet T , yjet pch T . z, yjet, pjet T , and collision centrality. The correction is estimated as a ratio of the UE charged particle yield evaluated by two different methods using the Pb+Pb MC samples. The ﬁrst estimate uses the cone method discussed above. The second method calculates the UE contribution in the data overlay MC samples from tracks, within the area of a jet, that do not have an associated generated primary particle. The size of the correction is less than 2% at low z or pch T where the UE has the largest impact, and has only a small dependence on pjet T . Here Ncone is the number of background cones used in the UE determination of a given jet, Ncone ch represents the number of charged particles summed over all background cones, and R represents the distance between the center of a cone and the direction of a given charged particle. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … z 2 − 10 1 − 10 1 [%] D(z) R δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < 0-10% [GeV] T p 1 10 [%] ) T D(p R δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < 0-10% FIG. 5. Summary of the systematic uncertainties for 0–10% central RD(z) (left) and RD(pT) (right) ratios, for jets in the 126–158 GeV pjet T interval. The systematic uncertainties due to JES, JER, unfolding, UE contribution, MC nonclosure, and tracking are shown along with the total systematic uncertainty from all sources. z 2 − 10 1 − 10 1 [%] D(z) R δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < 0-10% [GeV] T p 1 10 [%] ) T D(p R δ 20 − 15 − 10 − 5 − 0 5 10 15 20 25 JES JER Unfolding MC non-closure Tracking UE subtraction Total =0.4 R tk anti- \|<2.1 jet y \| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, -1 = 5.02 TeV, 25 pb s , pp < 158 GeV jet T p 126 < 0-10% FIG. 5. Summary of the systematic uncertainties for 0–10% central RD(z) (left) and RD(pT) (right) ratios, for jets in the 126–158 GeV pjet T interval. The systematic uncertainties due to JES, JER, unfolding, UE contribution, MC nonclosure, and tracking are shown along with the total systematic uncertainty from all sources. FIG. 5. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … The term ε(pch T , ηch) is the efﬁciency for reconstructing charged particles, estimated as a function of pch T and ηch without requiring track-to-jet matching. pT q g j g The estimated contribution from the UE in each cone is corrected for the difference in the average yield of UE charged particles at a given pch T between the η position of the cone and η position of the jet. This correction is based on the centrality-, pch T -, and η-dependent distribution of charged-particle yields in minimum-bias data events. An additional correction is T The contribution from fake tracks to the fragmentation functionsisestimatedfromtheMCsampleswithoutminimum- bias interactions overlaid. The fraction of these tracks is found to be below 2% of the tracks that pass the selection in all track and jet kinematic regions in this analysis. 024908-6 PHYSICAL REVIEW C 98, 024908 (2018) MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Summary of the systematic uncertainties for 0–10% central RD(z) (left) and RD(pT) (right) ratios, for jets in the 126–158 GeV pjet T interval. The systematic uncertainties due to JES, JER, unfolding, UE contribution, MC nonclosure, and tracking are shown along with the total systematic uncertainty from all sources. The UE distributions corrected for the additive contribution of fake tracks, d ˜nUE+fake ch /dpch T and d ˜nUE+fake ch /dz, are then subtracted from the measured distributions, and the subtracted charged-particle yields and fragmentation functions are evalu- ated: [44]. Using MC samples, four-dimensional response matrices are created using the generator-level and reconstructed pjet T , and the generator-level and reconstructed charged-particle z or pT. Separate unfolding matrices are constructed for pp data and each centrality interval in Pb+Pb collisions. A separate one-dimensional Bayesian unfolding is used to correct the measured pjet T spectra which are used to normalize the unfolded unnormalized fragmentation functions, dnunfolded ch /dpT and dnunfolded ch /dz. To achieve better agreement with the data, the MC jet spectra and fragmentation functions are reweighted to match the shapes in the reconstructed data. The Bayesian pro- cedure requires a choice in the number of iterations. Additional iterations reduce the sensitivity to the choice of prior, but may amplify statistical ﬂuctuations in the distributions. After four iterations for both the one-dimensional and two-dimensional unfoldings the fragmentation functions are stable for both the Pb+Pb and pp data. The ﬁnal, particle-level corrected distributions are deﬁned as dnsub ch dz = dnmeas ch dz −d ˜nUE+fake ch dz , Dsub(z) = 1 Nmeas jet dnsub ch dz , and dnsub ch dpch T = dnmeas ch dpch T −d ˜nUE+fake ch dpch T , Dsub pch T = 1 Nmeas jet dnsub ch dpch T , where Nmeas jet is the total number of measured jets in a given pjet T interval. The signal-to-background ratio, nsub ch /nUE ch , strongly depends on the collision centrality and pch T . Figure 2 shows the distributions prior to the UE and fake-track subtraction, dnmeas ch dpch T , divided by the distributions after the subtraction, dnsub ch dpch T , where Nmeas jet is the total number of measured jets in a given pjet T interval. The signal-to-background ratio, nsub ch /nUE ch , strongly depends on the collision centrality and pch T . MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Figure 2 shows the distributions prior to the UE and fake-track subtraction, dnmeas ch dpch T , divided by the distributions after the subtraction, dnsub ch dpch T , T T as a function of pch T for three centrality selections. In 0–10% central collisions, the distributions prior to subtraction are over ten times larger than the subtracted distributions for the most extreme case of 1 GeV charged particles. This ratio is reduced to approximately 2 in peripheral collisions at the same charged particle pT. The fake-track contribution to the fragmentation functions is subtracted from the measured fragmentation func- tions in both the pp and Pb+Pb collisions; the UE subtraction is performed only for the Pb+Pb measurement as the UE contribution is negligible in the pp collisions (less than 2% over the entire kinematic range measured). where Nunfolded jet is the unfolded number of jets in a given pjet T interval. The performance of the analysis procedure is tested by dividing the MC events in half and using one half to generate response matrices with which the other half is unfolded and the ratio of unfolded to generator-level fragmentation functions4 is evaluated. This procedure tests all the analysis corrections and the unfolding procedure. Good recovery of the generator-level (truth) MC distributions is observed for the unfolded events. The deviations from the exact recovery of the generator- level MC distributions, the nonclosure, are included in the systematic uncertainties. The ratios of Dsub(z) and Dsub(pch T ) distributions to the unfolded D(z) and D(pT) distributions are To remove the effects of bin migration due to the jet energy and track momentum resolution, the subtracted dnsub ch /dz and dnsub ch /dpch T distributions are corrected by using a two- dimensional Bayesian unfolding procedure [42] in z or pT and pjet T as implemented in the RooUnfold package [43]. Two-dimensionalunfoldingisusedbecausethecalorimetricjet energy response depends on the fragmentation pattern of the jet 4The generator-level fragmentation functions are constructed using generator-level jets and primary charged particles. 024908-7 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … In most cases, the statistical uncertainties are smaller than the marker size. shown in Fig. 3 for pp collisions and 0–10% central Pb+Pb collisions. The magnitude of the unfolding effect varies as a function of pjet T , pch T , and centrality. The effect of the unfolding is similar in pp and Pb+Pb collisions at low z and pT, but for higher-momentum particles within the jet, the effect of the unfolding in pp and Pb+Pb collisions differs by up to 25% between the two collision systems for 126 < pjet T < 158 GeV. This difference is due to UE ﬂuctuations, which lead to poorer jet energy resolution in Pb+Pb collisions than in pp collisions. With increasing pjet T , the effect of UE ﬂuctuations decreases; for 251 < pjet T < 316 GeV the effect of the unfolding is similar in Pb+Pb and pp collisions at all value of z and pT. The effect of the unfolding is larger at high z and pT due to the steepness of the fragmentation function near z = 1. The shaded boxes in Fig. 3 show the size of systematic uncertainties associated with the unfolding which originate from the sensitivity of the unfolding to the shape of input MC distributions, as described in the next section. With increasing pjet T , the effect of UE ﬂuctuations decreases; for 251 < pjet T < 316 GeV the effect of the unfolding is similar in Pb+Pb and pp collisions at all value of z and pT. The effect of the unfolding is larger at high z and pT due to the steepness of the fragmentation function near z = 1. The shaded boxes in Fig. 3 show the size of systematic uncertainties associated with the unfolding which originate from the sensitivity of the unfolding to the shape of input MC distributions, as described in the next section. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … z −2 10 −1 10 1 ) z ( D −4 10 −2 10 1 2 10 4 10 6 10 8 10 \| <2.1 jet y\| ATLAS = 5.02 TeV s , pp -1 25 pb =0.4 R tk anti- < 158 GeV x10 p 126 < < 200 GeV x10 p 158 < < 251 GeV x10 p 200 < < 316 GeV x10 p 251 < < 398 GeV x10 p 316 < [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D −6 10 −3 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS = 5.02 TeV s , pp -1 25 pb =0.4 R tk anti- < 158 GeV x10 p 126 < < 200 GeV x10 p 158 < < 251 GeV x10 p 200 < < 316 GeV x10 p 251 < < 398 GeV x10 p 316 < FIG. 6. Fragmentation functions, D(z) (left) and D(pT) (right), in pp collisions measured in ﬁve pjet T ranges from 126 to 398 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. z −2 10 −1 10 1 ) z ( D −4 10 −2 10 1 2 10 4 10 6 10 8 10 \| <2.1 jet y\| ATLAS = 5.02 TeV s , pp -1 25 pb =0.4 R tk anti- < 158 GeV x10 p 126 < < 200 GeV x10 p 158 < < 251 GeV x10 p 200 < < 316 GeV x10 p 251 < < 398 GeV x10 p 316 < [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D −6 10 −3 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS = 5.02 TeV s , pp -1 25 pb =0.4 R tk anti- < 158 GeV x10 p 126 < < 200 GeV x10 p 158 < < 251 GeV x10 p 200 < < 316 GeV x10 p 251 < < 398 GeV x10 p 316 < z FIG. 6. Fragmentation functions, D(z) (left) and D(pT) (right), in pp collisions measured in ﬁve pjet T ranges from 126 to 398 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … z −2 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 158 GeV jet T p 126 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p 1 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 158 GeV jet T p 126 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 7. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 126 to 158 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … [GeV] T p 1 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 158 GeV jet T p 126 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 z −2 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 158 GeV jet T p 126 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p FIG. 7. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 126 to 158 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. VI. SYSTEMATIC UNCERTAINTIES and pp collisions are re-evaluated. The difference between the varied and nominal distributions is used as an estimate of the resulting uncertainty. The following sources of systematic uncertainty are con- sidered: the jet energy scale (JES), the jet energy resolution (JER), the sensitivity of the unfolding to the prior, the residual nonclosure of the analysis procedure, UE contribution, and tracking-related uncertainties. For each variation accounting for a source of systematic uncertainty, the fragmentation functions and ratios of D(z) and D(pT) distributions in Pb+Pb The systematic uncertainty due to the JES in Pb+Pb collisions is composed of two parts: a centrality-independent baseline component and a centrality-dependent component. Only the centrality-independent baseline component is used in pp collisions; it is determined from in situ studies of the calorimeter response [37,45,46], and studies of the relative z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 251 GeV jet T p 200 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 251 GeV jet T p 200 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 9. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 200 to 251 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 200 GeV jet T p 158 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 200 GeV jet T p 158 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 8. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 158 to 200 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … 024908-8 PHYSICAL REVIEW C 98, 024908 (2018) MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 200 GeV jet T p 158 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 200 GeV jet T p 158 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 8. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 158 to 200 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. VI. SYSTEMATIC UNCERTAINTIES [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 251 GeV jet T p 200 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 251 GeV jet T p 200 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 9. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 200 to 251 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. 024908-9 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. VI. SYSTEMATIC UNCERTAINTIES [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 316 GeV jet T p 251 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 316 GeV jet T p 251 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 10. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 251 to 316 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. energy scale difference between the jet reconstruction pro- cedure in heavy-ion collisions [45] and the procedure in pp collisions [37]. The centrality-dependent uncertainty reﬂects a modiﬁcation of parton showers by the Pb+Pb environment. It is evaluated by comparing calorimeter pjet T and the sum of pT of tracks within the jet in data and MC simulation. The size of the centrality-dependent uncertainty in the JES reaches 0.5% in the most central collisions. Each component that contributes to the JES uncertainty is varied separately by ±1 standard deviation for each interval in pjet T , and the response matrix is recomputed accordingly. The data are unfolded with these matrices. The resulting uncertainty on the fragmentation functions increases with increasing z and particle pT at ﬁxed pjet T and decreases with increasing pjet T . VI. SYSTEMATIC UNCERTAINTIES The uncertainty in the fragmentation functions due to the JER is evaluated by repeating the unfolding procedure with modiﬁed response matrices, where an additional contribution is added to the resolution of the reconstructed pjet T using a [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 398 GeV jet T p 316 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 398 GeV jet T p 316 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 398 GeV jet T p 316 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 11. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 316 to 398 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. VI. SYSTEMATIC UNCERTAINTIES z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 316 GeV jet T p 251 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 [GeV] T p 1 10 2 10 ] -1 ) [GeV T p ( D 6 − 10 3 − 10 1 3 10 6 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 316 GeV jet T p 251 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 10. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 251 to 316 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. VI. SYSTEMATIC UNCERTAINTIES z 2 − 10 1 − 10 1 ) z ( D 4 − 10 1 − 10 2 10 5 10 8 10 \| <2.1 jet y\| ATLAS -1 = 5.02 TeV, 0.49 nb NN s Pb+Pb, =0.4 jets R tk anti- < 398 GeV jet T p 316 < 3 0 - 10% x 10 2 10 - 20% x 10 1 20 - 30% x 10 0 30 - 40% x 10 -1 40 - 60% x 10 -2 60 - 80% x 10 FIG. 11. Fragmentation functions, D(z) (left) and D(pT) (right), in Pb+Pb collisions measured in six different centrality classes for pjet T of 316 to 398 GeV. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. In most cases, the statistical uncertainties are smaller than the marker size. 024908-10 024908-10 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) FIG. 12. Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated for ﬁve pjet T r \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic trality decreases from top to bottom panels and pjet T increases from left to right panels. ussiansmearingprocedure.Thesmearingfactorisevaluated The unfolding uncertainty is estimated by ge FIG. 12. Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated for ﬁve pjet T ranges for jets with \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. Gaussiansmearingprocedure.Thesmearingfactorisevaluated using an in situ technique in 13 TeV pp data involving studies of dijet energy balance [47,48]. An additional uncertainty is included to account for differences between the heavy-ion- style jet reconstruction and that used in analyses of 13 TeV pp data. The size of the resulting uncertainty on the fragmentation functions due to the JER typically reaches 10% for the highest charged-particle z and pT bins and decreases with decreasing charged-particle z and pT at ﬁxed pjet T . The positive and negative uncertainties from the JER are symmetrized. The unfolding uncertainty is estimated by generating the response matrices from the MC distributions without reweight- ing in pjet T , D(z), and D(pT). VI. SYSTEMATIC UNCERTAINTIES An additional uncertainty is assigned for the nonclosure of the unfolded distributions in simulations, as described in Sec. V. The magnitude of the uncertainty due to the unfolding and the nonclosure is typically below 2% and 5%, respectively. The systematic uncertainty associated with the estimation of the UE contribution on the fragmentation functions has two components. First, the parameter that excludes random cones 024908-11 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. FIG. 13. Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated for ﬁve pjet T h \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic ntrality decreases from top to bottom panels and pjet T increases from left to right panels. FIG. 13. Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated for ﬁve pjet T ranges for jets with \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. alternatively evaluated using an efﬁciency-corrected differ- ential yield of charged particles d4nch/dηchdφchdpch T d, where is the difference in azimuth of the charged particle from the second-order event plane, evaluated in minimum-bias Pb+Pb events. To each event considered, a weight is assigned such that the event sample obtained from the minimum-bias trigger has the same centrality distribution as the sample collected by the jet trigger. The resulting uncertainty on the fragmentation functions is smaller than 10% at low z or pT and it rapidly decreases in higher charged-particle z or pT bins. from the estimate is varied. Random cones are assumed to be associated with a hard process and excluded if the centroid of the cone is R < 0.8 from a reconstructed jet with pT > 90 GeV. The exclusion requirement is changed to R < 1.2 to estimate the sensitivity of the UE contributions. The size of the resulting uncertainty on the fragmentation function is everywheresmallerthan3%anditdecreasesinhighercharged- particle z or pT. The second component of the UE uncertainty arises from a difference when the UE from the cone method is compared with an alternative UE estimation. The UE is 024908-12 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) IG. 14. VI. SYSTEMATIC UNCERTAINTIES Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T ra \|yjet\| < 0.3. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic u rality decreases from top to bottom panels and pjet T increases from left to right panels. he uncertainties related to the track reconstruction and i i i f l U i i l d evaluated by varying the material description. Th i i h k i fﬁi FIG. 14. Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T ranges for jets with \|yjet\| < 0.3. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. The uncertainties related to the track reconstruction and selection originate from several sources. Uncertainties related to the fake rate, the material description in simulation, and the track transverse momentum are obtained from studies in data and simulation described in Ref. [49]. The systematic uncertainty on the fake-track rate is 30% in both collision systems [49]. The contamination of fake tracks is less than 2%, and the resulting uncertainty on the fragmentation functions is at most 0.5%. The sensitivity of the tracking efﬁciency to the description of the inactive material in the MC samples is evaluated by varying the material description. This resulting uncertainty in the track reconstruction efﬁciency is between 0.5% and 2% over the track pT range used in the analysis. An additional uncertainty takes into account a possible residual misalignment of the tracking detectors in pp and Pb+Pb data- taking. The alignment in these data sets is checked in situ using Z →μ+μ−events, and a track-pT dependent uncertainty arises from the ﬁnite size of this sample. The resulting uncer- tainties on the fragmentation functions are typically smaller than 1%, except at large z, where they are as large as 4%. 024908-13 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. FIG. 15. Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T range < \|yjet\| < 2.1. VI. SYSTEMATIC UNCERTAINTIES The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic trality decreases from top to bottom panels and pjet T increases from left to right panels. additional uncertainty on the tracking efﬁciency due to the This variation affects the track reconstruction efﬁc FIG. 15. Ratios of D(z) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T ranges for jets with 1.2 < \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. An additional uncertainty on the tracking efﬁciency due to the high local track density in the core of jets is 0.4% [41] for all j An additional uncertainty on the tracking efﬁciency due to the high local track density in the core of jets is 0.4% [41] for all pjet T ranges in this analysis. The uncertainty due to the track selection is evaluated by repeating the analysis with an addi- tional requirement on the signiﬁcance of the distance of closest approach of the track to the primary vertex. This uncertainty affects the track reconstruction efﬁciencies, track momentum resolution, and rate of fake tracks. The resulting uncertainty typically varies from 1% at low track pT to 5% at high track pT. Finally, the track-to-particle matching requirements are varied. This variation affects the track reconstruction efﬁciency, track momentum resolution, and rate of fake tracks. The resulting systematic uncertainty in the fragmentation functions is less than 0.5%. Example systematic uncertainties on the D(z) and D(pT) distributions for jets in the 126–158 GeV pjet T range measured in the two collision systems are presented in Fig. 4. All track-related systematic uncertainties are added in quadrature and presented as a total tracking uncertainty. The systematic uncertainties from each source are assumed to be uncorrelated, 024908-14 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) . Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T < 0.3. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic decreases from top to bottom panels and pjet T increases from left to right panels. FIG. 16. VI. SYSTEMATIC UNCERTAINTIES Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T ranges for jets with \|yjet\| < 0.3. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. so they are combined in quadrature to obtain the total system- atic uncertainty. variations of the ratios from their central values are used as the correlated systematic uncertainty. The uncorrelated uncertain- ties are added in quadrature. Each systematic uncertainty is as- sumed to be fully correlated with itself between different rapid- ity bins. The systematic uncertainty from each source, except the nonclosure of the unfolded distributions and the residual misalignment of thetrackingdetectors, is bin-to-bincorrelated. The total systematic uncertainties of the RD(z) and RD(pT) distributions are shown in Fig. 5 for one selected pjet T range. The correlations between the various systematic compo- nents are considered in evaluating the ratios of Pb+Pb to pp fragmentation functions. The unfolding and the MC nonclo- sure are each taken to be uncorrelated between the two collision systems. All other uncertainties are taken to be correlated. For the correlated uncertainties, the ratios are re-evaluated by applying the variation to both collision systems; the resulting 024908-15 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. FIG. 17. Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four p with 1.2 < \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indic ertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. VII. RESULTS The D(z) and D(pT) distributions in pp c FIG. 17. Ratios of D(pT) distributions in six centrality intervals of Pb+Pb collisions to pp collisions evaluated in four pjet T ranges for jets with 1.2 < \|yjet\| < 2.1. The vertical bars on the data points indicate statistical uncertainties, while the shaded bands indicate systematic uncertainties. Centrality decreases from top to bottom panels and pjet T increases from left to right panels. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) FIG. 18. RD(z) (left) and RD(pT) (right) for 126–158 GeV jets for collision energies of 5.02 TeV (this analysis) and 2.76 TeV [16]. The vertical bars on the data points indicate statistical uncertainties while the boxes indicate systematic uncertainties. FIG. 18. RD(z) (left) and RD(pT) (right) for 126–158 GeV jets for collision energies of 5.02 TeV (this analysis) and 2.76 TeV [16]. The vertical bars on the data points indicate statistical uncertainties while the boxes indicate systematic uncertainties. The shapes of the RD(z) and RD(pT) distributions are similar for all centralities: inside the jets; the yields of particles with low pT or z are enhanced; there is a reduction for particles with intermediate pT or z; and the yields of particles with high pT or z are enhanced. This is qualitatively consistent with previous measurements of jet fragmentation at √sNN = 2.76 TeV [14–16]; a quantitative comparison is provided in Sec. VIII. The magnitudes of the deviations of the ratios from unity decrease with decreasing collision centrality. In the most central collisions, the size of the enhancement is as large as 70% at low pT or z and 30% at high pT or z. The depletion of charged-particle yields at intermediate pT and z is as large as 20%.Insomecentralityandpjet T rangesthereisadecreaseofthe fragmentation functions at the highest z values. In this region the statistical and systematic uncertainties are the largest; more precise measurements are needed to determine if a signiﬁcant decrease exists. Figures 14 and 15 show the RD(z) distributions for jets in the most central and most forward rapidity intervals, 0.0–0.3 and 1.2–2.1, respectively, for the six centrality intervals used in this analysis and for four pjet T intervals: 126–158, 158– 200, 200–251, and 251–316 GeV. Figures 16 and 17 show RD(pT) distributions for the same jet rapidity, centrality, and pjet T ranges. In all rapidity ranges, the RD(z) and RD(pT) distributions have the same qualitative shape and centrality dependence as the rapidity-inclusive results presented above. VII. RESULTS The D(z) and D(pT) distributions in pp collisions are shown in Fig. 6. The corresponding distributions in Pb+Pb collisions are shown in Figs. 7–11. In order to quantify the difference in the fragmentation functions between Pb+Pb and pp collisions, the ratios of D(z) and D(pT) distributions measured in Pb+Pb collisions to those measured in pp collisions, RD(z) and RD(pT), are shown in Figs. 12 and 13, respectively. In each ﬁgure, the shaded boxes indicate systematic uncertainties and the vertical bars show the statistical uncertainties. The D(z) and D(pT) distributions in pp collisions are shown in Fig. 6. The corresponding distributions in Pb+Pb collisions are shown in Figs. 7–11. In this section, results are presented of the measurement of the D(z) and D(pT) distributions for jet pT between 126 and 398 GeV and six centrality intervals in Pb+Pb collisions; the same distributions are presented in pp collisions for the same pjet T ranges. In order to study the effects of hot dense matter on the jet fragmentation process, ratios of Pb+Pb fragmentation functions to pp fragmentation functions are evaluated. In order to quantify the difference in the fragmentation functions between Pb+Pb and pp collisions, the ratios of D(z) and D(pT) distributions measured in Pb+Pb collisions to those measured in pp collisions, RD(z) and RD(pT), are shown in Figs. 12 and 13, respectively. In each ﬁgure, the shaded boxes indicate systematic uncertainties and the vertical bars show the statistical uncertainties. 024908-16 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … VIII. DISCUSSION The EQ model does not have a description of the soft processes from soft gluon radiation or the response of the hot QCD matter to the jet passing through it, so the comparison with data is only appropriate at z > 0.1. In order to determine how the fragmentation functions depend on pjet T , the fragmentation functions from three pjet T intervals are compared in Fig. 19. The D(pT) and D(z) distributions are closely related to each other, differing, pri- marily, in the normalization by pjet T in the deﬁnition of z [see Eq. (1)]. Therefore, a comparison of the modiﬁcations of the fragmentation functions as a function of pjet T can show whether the size of modiﬁcations scales with charged-particle z or with pT. The former would be expected for fragmentation effects, and the latter might indicate some scale in the QGP. The large pjet T range available in this measurement allows these two scenarios to be distinguished. Figure 19 shows that the excess of soft particles observed in central Pb+Pb collisions exhibits a much smaller pjet T dependence for the D(pT) ratios than for the D(z) ratios; the transition from enhancement to suppressionforsoftfragmentsoccursatpT around4GeVforall pjet T values investigated in this analysis. The same comparison can be made for the hard particles. In this case, Fig. 19 shows that the enhancement of hard fragments with z ≳0.3 is nearly independent of pjet T . Figure 21 shows a comparison between measured RD(pT) and the hybrid model calculation with Rres = 3 for three pjet T in- tervals.Themagnitudeoftheenhancementofhigh-pT particles in the calculation agrees with the observations for pjet T in the ranges 126–158 and 200–251 GeV. The RD(z) values are also compared in Fig. 22 with a third model which uses calculations based on soft collinear effective theory (SCET) [55,56]. This model well describes RD(z) in the low and intermediate z regions, but does not reproduce the enhancement in the high-z region observed in the data. The fragmentation functions have been calculated within a hybrid model of jet quenching, which uses perturbative techniques for the high-Q2 processes in jet evolution and strong coupling for the low momentum scales associated with the QGP [50,51]. VIII. DISCUSSION In this section, the results from the previous section are further discussed and compared to theoretical models. In order to make a direct comparison with measurements at 2.76 TeV, Fig. 18 overlays the RD(z) and RD(pT) distributions measured in 2.76 TeV collisions [16] on those obtained in this FIG. 19. RD(z) (left) and RD(pT) (right) ratios for three pjet T ranges: 126–158 GeV (circles), 200–251 GeV (diamonds), and 316–398 GeV (crosses). The statistical uncertainties are shown as bars and the systematic uncertainties as outlined boxes. FIG. 19. RD(z) (left) and RD(pT) (right) ratios for three pjet T ranges: 126–158 GeV (circles), 200–251 GeV (diamonds), and 316–398 GeV (crosses). The statistical uncertainties are shown as bars and the systematic uncertainties as outlined boxes. 024908-17 024908-17 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. FIG. 20. RD(z) forjetswith126 < pjet T < 158GeVcomparedwith calculations from Ref. [51] (hybrid model) for Rres = 0 (dot-dashed curve), Rres = 3 (dashed curve), and to calculations from Ref. [21] (EQ model). FIG. 20. RD(z) forjetswith126 < pjet T < 158GeVcomparedwith calculations from Ref. [51] (hybrid model) for Rres = 0 (dot-dashed curve), Rres = 3 (dashed curve), and to calculations from Ref. [21] (EQ model). FIG. 21. RD(pT) ratios for three pjet T ranges: 126–158 GeV (cir- cles), 200–251 GeV (diamonds), and 316–398 GeV (crosses) com- pared with calculations from the hybrid model [51] with Rres = 3. FIG. 21. RD(pT) ratios for three pjet T ranges: 126–158 GeV (cir- cles), 200–251 GeV (diamonds), and 316–398 GeV (crosses) com- pared with calculations from the hybrid model [51] with Rres = 3. FIG. 20. RD(z) forjetswith126 < pjet T < 158GeVcomparedwith calculations from Ref. [51] (hybrid model) for Rres = 0 (dot-dashed curve), Rres = 3 (dashed curve), and to calculations from Ref. [21] (EQ model). FIG. 21. RD(pT) ratios for three pjet T ranges: 126–158 GeV (cir- cles), 200–251 GeV (diamonds), and 316–398 GeV (crosses) com- pared with calculations from the hybrid model [51] with Rres = 3. analysisat5.02TeV.Thetwomeasurementsatthetwocollision energies quantitatively agree over the entire z and charged- particle pT range of the measurement; no signiﬁcant collision energy dependence is observed [the lowest point in the D(pT) ratios differs by less than two standard deviations when the statistical and systematic uncertainties are combined]. as Lres = Rres/πT where Rres is a parameter of the model. VIII. DISCUSSION The fragmentation functions measured here are compared with calculations from this model in Fig. 20 for two values of Rres. The calculations with Rres = 3 are qualitatively consistent with the measurement at high z and pT. At low z and pT, the results of the calculations are below the data, in agreement with prior observations in comparisons to related observables [52]. Also shown in Fig. 20 is a calculation from Ref. [21] which is a phenomenological model, the effective quenching (EQ) model, incorporating energy-loss effects through two downward shifts in the pjet T spectrum: one for quark-initiated jets and a larger one for gluon-initiated jets. In this case, the jets fragment as in vacuum, but RD(z) differs from unity due to an increase in the fraction of quark jets in Pb+Pb collisions relative to pp collisions at a ﬁxed pjet T . Since quark jets are more likely to produce high-z particles than gluon jets [53,54] this causes RD(z) > 1 at high z in the model predictions. The EQ model does not have a description of the soft processes from soft gluon radiation or the response of the hot QCD matter to the jet passing through it, so the comparison with data is only appropriate at z > 0.1. as Lres = Rres/πT where Rres is a parameter of the model. The fragmentation functions measured here are compared with calculations from this model in Fig. 20 for two values of Rres. The calculations with Rres = 3 are qualitatively consistent with the measurement at high z and pT. At low z and pT, the results of the calculations are below the data, in agreement with prior observations in comparisons to related observables [52]. Also shown in Fig. 20 is a calculation from Ref. [21] which is a phenomenological model, the effective quenching (EQ) model, incorporating energy-loss effects through two downward shifts in the pjet T spectrum: one for quark-initiated jets and a larger one for gluon-initiated jets. In this case, the jets fragment as in vacuum, but RD(z) differs from unity due to an increase in the fraction of quark jets in Pb+Pb collisions relative to pp collisions at a ﬁxed pjet T . Since quark jets are more likely to produce high-z particles than gluon jets [53,54] this causes RD(z) > 1 at high z in the model predictions. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) FIG. 22. RD(z) for three pjet T ranges: 126–158 GeV (circles), 200– 251 GeV (diamonds), and 316–398 GeV (crosses) compared with calculations from the SCET model [55,56]. The P ch T \|cent represents the total transverse momentum carried byparticlesinthelowpT enhancementregion.Thedependence of Nch\|cent and P ch T \|cent on pjet T and centrality is presented in Fig. 23. Overall, both quantities are found to increase as a function of pjet T and collision centrality. In the most central collisions, Nch increases from approximately 1.5 to 2.0 particles over the pjet T range of this measurement. The amount of transverse momentum carried by these particles increases from approximately 2.5 to 4 GeV over the same pjet T range. In peripheral collisions, the number of particles contributing to the enhancement is much smaller, approximately 0.2 particles carrying less than 0.5 GeV of transverse momentum in the lowest pjet T range. These results are in qualitative agreement with measurements of the same quantities in √sNN = 2.76 TeV Pb+Pb collisions [16]; however, the pjet T ranges are not the same as used in this analysis and the pjet T dependence is not reported in that measurement. p In order to quantify the rapidity dependence, the ratio of RD(z) in the rapidity intervals 0.3–0.8, 0.8–1.2, and 1.2–2.1 to the RD(z) in \|yjet\| < 0.3 is shown in Fig. 24 for pjet T intervals of 126–158, 158–200, and 200–251 GeV and for 0–10%, 10–20%, and 20–30% central collisions. A similar quantity was reported in Ref. [16] for 100–398 GeV jets at 2.76 TeV. In that measurement, a small rapidity dependence for RD(z) is observed at high z for jets with \|yjet\| < 0.8; however, no strong conclusion could be drawn due to the size of the uncertainties. The pjet T intervals used in the measurement presented here are selected to be similar to those used in the measurement of frag- mentation functions at 2.76 TeV. Furthermore, jets populating the 200–251 GeV pjet T interval in collisions at 5.02 TeV have similar fractions of quark- and gluon-initiated jets as jets hav- ing pT between 126 and 158 GeV in 2.76 TeV collisions. The ratiosofRD(z) evaluatedinvariousrapidityintervalstothemost central rapidity RD(z) in different pjet T intervals suggest with a FIG. 22. VIII. DISCUSSION Within this model, there is a length scale Lreswhich can be interpreted as the minimum distance required to resolve a parton as separate from the others in the showering process when it occurs in the QGP medium. The scale Lres can be expressed in terms of the temperature of QGP, T , In order to quantify the magnitude of the low-pT enhance- ment in the D(pT) distributions in Pb+Pb collisions compared to pp collisions, the difference between the two distributions is evaluated for the pjet T and centrality intervals used in this 024908-18 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … RD(z) for three pjet T ranges: 126–158 GeV (circles), 200– 251 GeV (diamonds), and 316–398 GeV (crosses) compared with calculations from the SCET model [55,56]. analysis: Nch\|cent ≡ pT,max pT,min [D(pT)\|cent −D(pT)\|pp]dpT, where “cent” represents one of the six centrality intervals, and the values of pT,min and pT,max are boundaries of the low pT enhancement region, chosen to be 1.0 and 4.2 GeV, respectively. In addition, the pT-weighted difference between the same quantities is also computed: P ch T cent ≡ pT,max pT,min [D(pT)\|cent −D(pT)\|pp]pTdpT. T cent pT,min [ (pT)\|cent (pT)\|pp]pT pT central rapidity RD(z) in different pT intervals suggest with a FIG. 23. Difference between Pb + Pb collisions and pp collisions in the total yield of charged particles Nch\|cent (left), and difference in the total transverse momentum carried by charged particles P ch T \|cent (right) for particles with pT from 1 < pT < 4.2 GeV evaluated as a function of pjet T for six centrality intervals. The vertical bars on the data points indicate statistical uncertainties while the boxes indicate systematic uncertainties. FIG. 23. Difference between Pb + Pb collisions and pp collisions in the total yield of charged particles Nch\|cent (left), and difference in the total transverse momentum carried by charged particles P ch T \|cent (right) for particles with pT from 1 < pT < 4.2 GeV evaluated as a function of pjet T for six centrality intervals. The vertical bars on the data points indicate statistical uncertainties while the boxes indicate systematic uncertainties. 024908-19 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. FIG. 24. Ratio of the rapidity-selected RD(z) distributions to the RD(z) distributions measured in \|yjet\| < 0.3 for three pjet T ranges and three centrality intervals. The vertical bars on the data points indicate statistical uncertainties while the shaded bands indicate systematic uncertainties. FIG. 24. Ratio of the rapidity-selected RD(z) distributions to the RD(z) distributions measured in \|yjet\| < 0.3 for three pjet T ranges and three centrality intervals. The vertical bars on the data points indicate statistical uncertainties while the shaded bands indicate systematic uncertainties. FIG. 25. Comparison of the measured ratio of the rapidity-selected RD(z) distributions to the RD(z) distributions measured in \|yjet\| < 0.3 and the same quantity evaluated in the hybrid model [51] for Rres = 3 and in the EQ model [21]. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … momentum and rapidity dependence of the modiﬁcations to jet fragmentation in Pb+Pb collisions and, together with other jet measurements in heavy-ion collisions, will constrain models of jet quenching in the QGP created in heavy-ion collisions. low signiﬁcance a small enhancement of yields of fragments with low and intermediate z and reduction of yields of high-z fragments for more forward jets in the most central Pb + Pb collisions. However, the observation for high-z fragments is of limited signiﬁcance due to the limited size of the available data sample. Figure 25 shows the same ratios for the 0–10% centrality interval compared with calculations from the hybrid model [51] and the effective quenching model [21]. Both cal- culations are consistent with the data for jets with \|yjet\| < 1.2 with larger deviations in rapidity interval 1.2 < \|yjet\| < 2.1. ACKNOWLEDGMENTS We thank CERN for the very successful operation of the LHC, as well as the support staff from our institutions without whom ATLAS could not be operated efﬁciently. We acknowl- edge the support of ANPCyT, Argentina; YerPhI, Armenia; ARC, Australia; BMWFW and FWF, Austria; ANAS, Azer- baijan; SSTC, Belarus; CNPq and FAPESP, Brazil; NSERC, NRC and CFI, Canada; CERN; CONICYT, Chile; CAS, MOST and NSFC, China; COLCIENCIAS, Colombia; MSMT CR, MPO CR and VSC CR, Czech Republic; DNRF and DNSRC, Denmark; IN2P3-CNRS, CEA-DRF/IRFU, France; SRNSFG, Georgia; BMBF, HGF, and MPG, Germany; GSRT, Greece; RGC, Hong Kong SAR, China; ISF, I-CORE and Benoziyo Center, Israel; INFN, Italy; MEXT and JSPS, Japan; CNRST, Morocco; NWO, Netherlands; RCN, Nor- way; MNiSW and NCN, Poland; FCT, Portugal; MNE/IFA, Romania; MES of Russia and NRC KI, Russian Federation; JINR; MESTD, Serbia; MSSR, Slovakia; ARRS and MIZŠ, Slovenia; DST/NRF, South Africa; MINECO, Spain; SRC and Wallenberg Foundation, Sweden; SERI, SNSF and Cantons of Bern and Geneva, Switzerland; MOST, Taiwan; TAEK, Turkey; STFC, United Kingdom; DOE and NSF, United States of America. In addition, individual groups and members have received support from BCKDF, the Canada Council, CANARIE, CRC, Compute Canada, FQRNT, and the Ontario Innovation Trust, Canada; EPLANET, ERC, ERDF, FP7, Horizon 2020 and Marie Skłodowska-Curie Actions, Euro- pean Union; Investissements d’Avenir Labex and Idex, ANR, Région Auvergne and Fondation Partager le Savoir, France; DFG and AvH Foundation, Germany; Herakleitos, Thales and Aristeia programmes co-ﬁnanced by EU-ESF and the Greek NSRF; BSF, GIF and Minerva, Israel; BRF, Norway; CERCA Programme Generalitat de Catalunya, Generalitat Valenciana, Spain; the Royal Society and Leverhulme Trust, United King- dom. The crucial computing support from all WLCG partners is acknowledged gratefully, in particular from CERN, the ATLAS Tier-1 facilities at TRIUMF (Canada), NDGF (Den- mark, Norway, Sweden), CC-IN2P3 (France), KIT/GridKA (Germany), INFN-CNAF (Italy), NL-T1 (Netherlands), PIC (Spain), ASGC (Taiwan), RAL (UK), and BNL (USA), the Tier-2 facilities worldwide, and large non-WLCG resource providers. Major contributors of computing resources are listed in Ref. [57]. [1] G. Roland, K. Šafaˇrík, and P. Steinberg, Heavy-ion collisions at the LHC, Prog. Part. Nucl. Phys. 77, 70 (2014). [2] W. Busza, K. Rajagopal, and W. van der Schee, Heavy ion collisions: The big picture, and the big questions, arXiv:1802.04801. [3] ALICE Collaboration, Measurement of charged jet suppression in Pb-Pb collisions at √sNN = 2.76 TeV, J. High Energy Phys. 03 (2014) 013. 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The vertical bars on the data points indicate statistical uncertainties while the shaded bars indicate systematic uncertainties. The band represents the statistical uncertainty of the calculations. FIG. 25. Comparison of the measured ratio of the rapidity-selected RD(z) distributions to the RD(z) distributions measured in \|yjet\| < 0.3 and the same quantity evaluated in the hybrid model [51] for Rres = 3 and in the EQ model [21]. The comparison with the hybrid model is done for three pjet T ranges in 0–10% central collisions. The comparison with the EQ model is shown for 126–158 GeV pjet T interval. The vertical bars on the data points indicate statistical uncertainties while the shaded bars indicate systematic uncertainties. 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SUMMARY This paper presents an analysis of 0.49 nb−1 of Pb+Pb and 25 pb−1 of pp collisions at √sNN = 5.02 TeV using data collected with the ATLAS detector at the LHC in 2015. The analysis measures the fragmentation functions of jets into charged particles and the distributions of charged-particle transverse momenta within R = 0.4 anti-kt jets with \|yjet\| < 2.1 and with pjet T from 126 to 398 GeV. The studies are performed as a function of the event centrality, jet rapidity, and jet transverse momentum for charged particles with transverse momentum greater than 1 GeV. g Centrality-dependent modiﬁcations to these fragmentation functions in Pb+Pb collisions are observed when compared with those measured in pp collisions. The magnitude of these modiﬁcations increases with increasing collision centrality. The ratios of fragmentation functions evaluated in Pb+Pb collisions to those in pp collisions exhibit enhancements both for transverse momentum less than 4 GeV and for z ≳0.3. Between these two enhancements there is a suppression of the fragmentation functions in Pb+Pb collisions compared to pp collisions. The enhancement of yields of low and high transverse momentum fragments is as large as 70% and 30%, respectively, in central collisions. The depletion of fragment yields with intermediate pT and z is as large as 20%. The difference in charged-particle multiplicity and total transverse momentum in Pb+Pb compared to pp collisions for 1.0 < pT < 4.2 GeV range increases with increasing centrality and jet transverse momentum. No signiﬁcant dependence of the high-z enhancement on the transverse momentum of the jet is observed. The SCET model describes the low pT excess and the EQ and hybrid models describe the high-z excess, but none of the models describes the modiﬁcation of the full fragmentation functions. A small increase in the modiﬁcation of yields of fragments with low and intermediate z is observed in forward jets compared to those at central rapidity. These measurementsprovidenewinformationaboutthejettransverse [4] ATLAS Collaboration, Measurements of the Nuclear Modiﬁca- tion Factor for Jets in Pb+Pb Collisions at √sNN = 2.76 TeV with the ATLAS Detector, Phys. Rev. Lett. 114, 072302 (2015). [5] CMS Collaboration, Measurement of inclusive jet cross sections in pp and PbPb collisions at √sNN = 2.76 TeV, Phys. Rev. C 96, 015202 (2017). [6] ATLAS Collaboration, Observation of a Centrality-Dependent Dijet Asymmetry in Lead-Lead Collisions at √sNN = 2.77 TeV 024908-21 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. IX. SUMMARY [22] ATLASCollaboration,Measurementofjetfragmentationin5.02 TeV proton–lead and proton–proton collisions with the ATLAS detector, Nucl. Phys. A 978, 65 (2018). [23] ATLAS Collaboration, Measurement of the charged-particle multiplicity inside jets from √s = 8 TeV pp collisions with the ATLAS detector, Eur. Phys. J. C 76, 322 (2016). 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Ferrari,169 P. Ferrari,118 R. Ferrari,68a D. E. Ferre y g D. Ferrere,52 C. Ferretti,103 F. Fiedler,97 A. Filipˇciˇc,89 F. Filthaut,117 K. D. Finelli,25 M. C. N. Fiol , , , p , , , , , C. Fischer,14 W. C. Fisher,104 N. Flaschel,44 I. Fleck,148 P. Fleischmann,103 R. R. M. Fletcher,133 T. Flick,179 B. M. Flierl,112 L. M. Flores,133 L. R. Flores Castillo,61a N. Fomin,17 G. T. Forcolin,98 A. Formica,142 F. A. Förster,14 A. C. Forti,98 C. Fischer,14 W. C. Fisher,104 N. Flaschel,44 I. Fleck,148 P. Fleischmann,103 R. R. M. Fletcher,133 T. Flick,179 B. M. Flierl,112 L. M. Flores,133 L. R. Flores Castillo,61a N. Fomin,17 G. T. Forcolin,98 A. Formica,142 F. A. Förster,14 A. C. Forti,98 W. S. Freund,78b D. Froidevaux,35 J. A. Frost,131 C. Fukunaga,161 E. Fullana Torregrosa,171 T. Fusayasu,114 J. Fuster,171 O. Gabizon,157 A. Gabrielli,23b,23a A. Gabrielli,18 G. P. Gach,81a S. Gadatsch,52 P. Gadow,113 G. Gagliardi,53b,53a L. G. Gagnon,107 C. Galea,27b B. Galhardo,136a,136c E. J. Gallas,131 B. J. Gallop,141 P. Gallus,138 G. Galster,39 R. Gamboa Goni,90 K. K. Gan,122 S. Ganguly,177 Y. Gao,88 Y. S. Gao,150,g C. García,171 J. E. García Navarro,171 J. A. García Pascual,15a M. Garcia-Sciveres,18 R. W. Gardner,36 N. Garelli,150 V. Garonne,130 K. Gasnikova,44 W. S. Freund,78b D. Froidevaux,35 J. A. Frost,131 C. Fukunaga,161 E. Fullana Torregrosa,171 T. Fusayasu,114 J. Fuster,171 O. Gabizon,157 A. Gabrielli,23b,23a A. Gabrielli,18 G. P. Gach,81a S. Gadatsch,52 P. Gadow,113 G. Gagliardi,53b,53a L. G. Gagnon,107 C. Galea,27b B. Galhardo,136a,136c E. J. Gallas,131 B. J. Gallop,141 P. Gallus,138 G. Galster,39 R. Gamboa Goni,90 K. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … K. Gan,122 S. Ganguly,177 Y. Gao,88 Y. S. Gao,150,g C. García,171 J. E. García Navarro,171 J. A. García Pascual,15a M. Garcia-Sciveres,18 R. W. Gardner,36 N. Garelli,150 V. Garonne,130 K. Gasnikova,44 53b 53 68 108 109 172 24 10 141 J. Geisen,51 M. Geisen,97 M. P. Geisler,59a K. Gellerstedt,43a,43b C. Gemme,53b M. H. Genest,56 C. Geng,103 S. Gentile,70a,70b C. Gentsos,159 S. George,91 D. Gerbaudo,14 G. Gessner,45 S. Ghasemi,148 M. Ghasemi Bostanabad,173 M. Ghneimat,24 B. Giacobbe,23b S. Giagu,70a,70b N. Giangiacomi,23b,23a P. Giannetti,69a A. Giannini,67a,67b S. M. Gibson,91 M. Gignac,143 D. Gillberg,33 G. Gilles,179 D. M. Gingrich,3,e M. P. Giordani,64a,64c F. M. Giorgi,23b P. F. Giraud,142 P. Giromini,57 G. Giugliarelli,64a,64c D. Giugni,66a F. Giuli,131 M. Giulini,59b S. Gkaitatzis,159 I. Gkialas,9,t E. L. Gkougkousis,14 P. Gkountoumis,10 L. K. Gladilin,111 C. Glasman,96 J. Glatzer,14 P. C. F. Glaysher,44 A. Glazov,44 M. Goblirsch-Kolb,26 y J. Godlewski,82 S. Goldfarb,102 T. Golling,52 D. Golubkov,140 A. Gomes,136a,136b,136d R. Goncalves Gama,78a R. Gonçalo,136a G G ll 50 L G ll 21 A G d 77 F G ll 21 J L G ki 57 S G ál d l H 171 S G l S ill 52 L. Goossens,35 P. A. Gorbounov,109 H. A. Gordon,29 B. Gorini,35 E. Gorini,65a,65b A. Gorišek,89 A. T. Goshaw,47 C. Gössling,45 M. I. Gostkin,77 C. A. Gottardo,24 C. R. Goudet,128 D. Goujdami,34c A. G. Goussiou,145 N. Govender,32b,u C. Goy,5 E. Gozani,157 I. Grabowska-Bold,81a P. O. J. Gradin,169 E. C. Graham,88 J. Gramling,168 E. Gramstad,130 S. Grancagnolo,19 V. Gratchev,134 P. M. Gravila,27f C. Gray,55 H. M. Gray,18 Z. D. Greenwood,93,v C. Grefe,24 K. Gregersen,92 I. M. Gregor,44 P. Grenier,150 K. Grevtsov,44 J. Grifﬁths,8 A. A. Grillo,143 K. Grimm,150 S. Grinstein,14,w Ph. Gris,37 J.-F. Grivaz,128 S. Groh,97 E. Gross,177 J. Grosse-Knetter,51 G. C. Grossi,93 Z. J. Grout,92 C. Grud,103 A. Grummer,116 L. Guan,103 W. Guan,178 J. Guenther,35 A. Guerguichon,128 F. Guescini,165a D. Guest,168 R. Gugel,50 B. Gui,122 T. Guillemin,5 S. Guindon,35 U. Gul,55 C. Gumpert,35 J. Guo,58c W. Guo,103 Y. Guo,58a,x Z. Guo,99 R. Gupta,41 S. Gurbuz,12c G. Gustavino,124 B. J. Gutelman,157 P. Gutierrez,124 C. Gutschow,92 C. Guyot,142 M. P. Guzik,81a C. Gwenlan,131 C. B. Gwilliam,88 A. Haas,121 C. Haber,18 8 34 58 24 166 181 1 4 125 H. K. Hadavand,8 N. Haddad,34e A. Hadef,58a S. Hageböck,24 M. Hagihara,166 H. Hakobyan,181,a M. Haleem,174 J. Haley,125 G. Halladjian,104 G. D. Hallewell,99 K. Hamacher,179 P. Hamal,126 K. Hamano,173 A. Hamilton,32a G. N. Hamity,146 H. K. Hadavand,8 N. C. Issever,131 S. Istin,12c,ac F. Ito,166 J. M. Iturbe Ponce,61a R. Iuppa,73a,73b A. Ivina,177 H. Iwasaki,79 J. M. Izen,42 V. Izzo,67a S. Jabbar,3 P. Jacka,137 P. Jackson,1 R. M. Jacobs,24 V. Jain,2 G. Jäkel,179 K. B. Jakobi,97 K. Jakobs,50 S. Jakobsen,74 C. Issever,131 S. Istin,12c,ac F. Ito,166 J. M. Iturbe Ponce,61a R. Iuppa,73a,73b A. Ivina,177 H. Iwasaki,79 J. M. Izen,42 V. Izzo,67a S. Jabbar,3 P. Jacka,137 P. Jackson,1 R. M. Jacobs,24 V. Jain,2 G. Jäkel,179 K. B. Jakobi,97 K. Jakobs,50 S. Jakobsen,74 T. Jakoubek,137 D. O. Jamin,125 D. K. Jana,93 R. Jansky,52 J. Janssen,24 M. Janus,51 P. A. Janus,81a G. Jarlskog,94 N. Javadov,77,d T. Jav˚urek,50 M. Javurkova,50 F. Jeanneau,142 L. Jeanty,18 J. Jejelava,156a,ad A. Jelinskas,175 P. Jenni,50,ae J. Jeong,44 S. Jézéquel,5 H. Ji,178 J. Jia,152 H. Jiang,76 Y. Jiang,58a Z. Jiang,150 S. Jiggins,50 F. A. Jimenez Morales,37 J. Jimenez Pena,171 S. Jin,15c A. Jinaru,27b O. Jinnouchi,162 H. Jivan,32c P. Johansson,146 K. A. Johns,7 C. A. Johnson,63 W. J. Johnson,145 K. Jon-And,43a,43b R. W. L. Jones,87 S. D. Jones,153 S. Jones,7 T. J. Jones,88 J. Jongmanns,59a P. M. Jorge,136a,136b J. Jovicevic,165a X. Ju,178 J. J. Junggeburth,113 A. Juste Rozas,14,w A. Kaczmarska,82 M. Kado,128 H. Kagan,122 M. Kagan,150 T. Kaji,176 E. Kajomovitz,157 C. W. Kalderon,94 A. Kaluza,97 S. Kama,41 A. Kamenshchikov,140 L. Kanjir,89 Y. Kano,160 V. A. Kantserov,110 J. Kanzaki,79 B. Kaplan,121 L. S. Kaplan,178 D. Kar,32c M. J. Kareem,165b E. Karentzos,10 S. N. Karpov,77 Z. M. Karpova,77 V. Kartvelishvili,87 A. N. Karyukhin,140 K. Kasahara,166 L. Kashif,178 R. D. Kass,122 A. Kastanas,151 Y. Kataoka,160 C. Kato,160 J. Katzy,44 K. Kawade,80 K. Kawagoe,85 T. Kawamoto,160 G. Kawamura,51 E. F. Kay,88 V. F. Kazanin,120b,120a R. Keeler,173 R. Kehoe,41 J. S. Keller,33 E. Kellermann,94 J. J. Kempster,21 J. Kendrick,21 O. Kepka,137 S. Kersten,179 B. P. Kerševan,89 R. A. Keyes,101 M. Khader,170 F. Khalil-Zada,13 A. Khanov,125 A. G. Kharlamov,120b,120a T. Kharlamova,120b,120a A. Khodinov,163 T. J. Khoo,52 E. Khramov,77 J. Khubua,156b S. Kido,80 M. Kiehn,52 C. R. Kilby,91 S. H. Kim,166 Y. K. Kim,36 N. Kimura,64a,64c O. M. Kind,19 B. T. King,88 D. Kirchmeier,46 J. Kirk,141 A. E. Kiryunin,113 T. Kishimoto,160 D. Kisielewska,81a V. Kitali,44 O. Kivernyk,5 E. Kladiva,28b T. Klapdor-Kleingrothaus,50 M. H. Klein,103 M. Klein,88 U. Klein,88 K. Kleinknecht,97 P. Klimek,119 A. Klimentov,29 R. Klingenberg,45,a T. Klingl,24 T. Klioutchnikova,35 F. F. Klitzner,112 P. Kluit,118 S. Kluth,113 E. Kneringer,74 E. B. F. G. Knoops,99 A. Knue,50 A. Kobayashi,160 D. Kobayashi,85 T. Kobayashi,160 M. Kobel,46 M. Kocian,150 P. Kodys,139 T. Koffas,33 E. Koffeman,118 N. M. Köhler,113 T. Koi,150 M. Kolb,59b I. Koletsou,5 T. Kondo,79 N. Kondrashova,58c K. Köneke,50 A. C. König,117 T. Kono,79 R. Konoplich,121,af V. Konstantinides,92 N. Konstantinidis,92 B. Konya,94 R. Kopeliansky,63 S. Koperny,81a K. Korcyl,82 K. Kordas,159 A. Korn,92 I. Korolkov,14 E. V. Korolkova,146 O. Kortner,113 S. Kortner,113 T. Kosek,139 V. V. Kostyukhin,24 A. Kotwal,47 A. Koulouris,10 A. Kourkoumeli-Charalampidi,68a,68b C. Kourkoumelis,9 E. Kourlitis,146 V. Kouskoura,29 A. B. Kowalewska,82 R. Kowalewski,173 T. Z. Kowalski,81a C. Kozakai,160 W. Kozanecki,142 A. S. Kozhin,140 V. A. Kramarenko,111 G. Kramberger,89 D. Krasnopevtsev,110 M. W. Krasny,132 A. Krasznahorkay,35 D. Krauss,113 J. A. Kremer,81a J. Kretzschmar,88 P. Krieger,164 K. Krizka,18 K. Kroeninger,45 H. Kroha,113 J. Kroll,137 J. Kroll,133 J. Krstic,16 U. Kruchonak,77 H. Krüger,24 N. Krumnack,76 M. C. Kruse,47 T. Kubota,102 S. Kuday,4b J. T. Kuechler,179 S. Kuehn,35 A. Kugel,59a F. Kuger,174 T. Kuhl,44 V. Kukhtin,77 R. Kukla,99 Y. Kulchitsky,105 S. Kuleshov,144b Y. P. Kulinich,170 M. Kuna,56 T. Kunigo,83 A. Kupco,137 T. Kupfer,45 O. Kuprash,158 H. Kurashige,80 L. L. Kurchaninov,165a Y. A. Kurochkin,105 M. G. Kurth,15d E. S. Kuwertz,173 M. Kuze,162 J. Kvita,126 T. Kwan,101 A. La Rosa,113 J. L. La Rosa Navarro,78d L. La Rotonda,40b,40a F. La Ruffa,40b,40a C. Lacasta,171 F. Lacava,70a,70b J. Lacey,44 D. P. J. Lack,98 H. Lacker,19 D. Lacour,132 E. Ladygin,77 R. Lafaye,5 B. Laforge,132 T. Lagouri,32c S. Lai,51 S. Lammers,63 W. Lampl,7 E. Lançon,29 U. Landgraf,50 M. P. J. Landon,90 M. C. Lanfermann,52 V. S. Lang,44 J. C. Lange,14 R. J. Langenberg,35 A. J. Lankford,168 F. Lanni,29 K. Lantzsch,24 A. Lanza,68a A. Lapertosa,53b,53a S. Laplace,132 J. F. Laporte,142 T. Lari,66a F. Lasagni Manghi,23b,23a M. Lassnig,35 T. S. Lau,61a A. Laudrain,128 M. Lavorgna,67a,67b A. T. Law,143 P. Laycock,88 M. Lazzaroni,66a,66b B. Le,102 O. Le Dortz,132 E. Le Guirriec,99 E. P. Le Quilleuc,142 M. LeBlanc,7 T. LeCompte,6 F. Ledroit-Guillon,56 C. A. Lee,29 G. R. Lee,144a L. Lee,57 S. C. Lee,155 B. Lefebvre,101 M. Lefebvre,173 F. Legger,112 C. Leggett,18 N. Lehmann,179 G. Lehmann Miotto,35 W. A. Leight,44 A. Leisos,159,ag M. A. L. Leite,78d R. Leitner,139 D. Lellouch,177 B. Lemmer,51 K. J. C. Leney,92 T. Lenz,24 B. Lenzi,35 R. Leone,7 S. Leone,69a C. Leonidopoulos,48 G. Lerner,153 C. Leroy,107 R. Les,164 A. A. J. Lesage,142 C. G. Lester,31 M. Levchenko,134 J. Levêque,5 D. Levin,103 L. J. Levinson,177 D. Lewis,90 B. Li,103 C-Q. Li,58a H. Li,58b L. Li,58c Q. Li,15d Q. Y. Li,58a S. Li,58d,58c X. Li,58c Y. Li,148 Z. Liang,15a B. Liberti,71a A. Liblong,164 K. Lie,61c S. Liem,118 A. Limosani,154 C. Y. Lin,31 K. Lin,104 T. H. Lin,97 R. A. Linck,63 B. E. Lindquist,152 A. L. Lionti,52 E. Lipeles,133 A. Lipniacka,17 M. Lisovyi,59b T. M. Liss,170,ah A. Lister,172 A. M. Litke,143 J. D. Little,8 B. Liu,76 B. L Liu,6 H. B. Liu,29 H. Liu,103 J. B. Liu,58a J. K. K. Liu,131 K. Liu,132 M. Liu,58a P. Liu,18 Y. Liu,15a Y. L. Liu,58a Y. W. Liu,58a M. Livan,68a,68b A. Lleres,56 J. Llorente Merino,15a S. L. Lloyd,90 C. Y. Lo,61b F. Lo Sterzo,41 E. M. Lobodzinska,44 P. Loch,7 A. Loesle,50 K. M. Loew,26 T. Lohse,19 K. Lohwasser,146 M. Lokajicek,137 B. A. Long,25 J. D. Long,170 R. E. Long,87 L. Longo,65a,65b K. A. Looper,122 J. A. Lopez,144b I. Lopez Paz,14 A. Lopez Solis,146 J. Lorenz,112 N. Lorenzo Martinez,5 M. Losada,22 P. J. Lösel,112 X. Lou,44 X. Lou,15a A. Lounis,128 J. Love,6 P. A. Love,87 J. J. Lozano Bahilo,171 H. Lu,61a M. Lu,58a N. Lu,103 Y. J. Lu,62 H. J. Lubatti,145 C. Luci,70a,70b A. Lucotte,56 C. Luedtke,50 F. Luehring,63 I. Luise,132 W. Lukas,74 L. Luminari,70a B. Lund-Jensen,151 M. S. Lutz,100 P. M. Luzi,132 D. Lynn,29 R. Lysak,137 E. Lytken,94 F. Lyu,15a V. Lyubushkin,77 H. Ma,29 L. L. Ma,58b Y. Ma,58b G. Maccarrone,49 A. Macchiolo,113 C. M. Macdonald,146 J. Machado Miguens,133,136b D. Madaffari,171 R. Madar,37 W. F. Mader,46 A. Madsen,44 N. Madysa,46 J. Maeda,80 K. Maekawa,160 S. Maeland,17 T. Maeno,29 A. S. Maevskiy,111 V. Magerl,50 C. Maidantchik,78b T. Maier,112 A. Maio,136a,136b,136d O. Majersky,28a S. Majewski,127 Y. Makida,79 N. Makovec,128 B. Malaescu,132 Pa. Malecki,82 V. P. Maleev,134 F. Malek,56 U. Mallik,75 D. Malon,6 C. Malone,31 S. Maltezos,10 S. Malyukov,35 J. Mamuzic,171 G. Mancini,49 I. Mandi´c,89 J. Maneira,136a L. Manhaes de Andrade Filho,78a J. Manjarres Ramos,46 K. H. Mankinen,94 A. Mann,112 A. Manousos,74 B. Mansoulie,142 J. D. Mansour,15a M. Mantoani,51 S. Manzoni,66a,66b G. Marceca,30 L. March,52 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … g A. C. Martyniuk,92 A. Marzin,35 L. Masetti,97 T. Mashimo,160 R. Mashinistov,108 J. Masik,98 A. L. Maslennikov,120b,120a L H Mason 102 L Massa 71a,71b P Mastrandrea 5 A Mastroberardino 40b,40a T Masubuchi 160 P Mättig 179 J Maurer 27b B. Maˇcek,89 S. J. Maxﬁeld,88 D. A. Maximov,120b,120a R. Mazini,155 I. Maznas,159 S. M. Mazza,143 N. C. Mc Fadden,116 G. Mc Goldrick,164 S. P. Mc Kee,103 A. McCarn,103 T. G. McCarthy,113 L. I. McClymont,92 E. F. McDonald,102 J. A. Mcfayden,35 G. Mchedlidze,51 M. A. McKay,41 K. D. McLean,173 S. J. McMahon,141 P. C. McNamara,102 175 173 32 100 145 50 112 y y g J. U. Mjörnmark,94 T. Mkrtchyan,181 M. Mlynarikova,139 T. Moa,43a,43b K. Mochizuki,107 P. Mogg,50 S. Mohapatra,38 S. Molander,43a,43b R. Moles-Valls,24 M. C. Mondragon,104 K. Mönig,44 J. Monk,39 E. Monnier,99 A. Montalbano,149 H. D. N. Nguyen,99 T. Nguyen Manh,107 E. Nibigira,37 R. B. Nickerson,131 R. Nicolaidou,142 J. Nielsen,143 N. Nikiforou,11 V. Nikolaenko,140,ak I. Nikolic-Audit,132 K. Nikolopoulos,21 P. Nilsson,29 Y. Ninomiya,79 A. Nisati,70a N. Nishu,58c R. Nisius,113 I. Nitsche,45 T. Nitta,176 T. Nobe,160 Y. Noguchi,83 M. Nomachi,129 I. Nomidis,132 M. A. Nomura,29 T. Nooney,90 35 131 89 46 138 126 168 92 M. Nordberg, N. Norjoharuddeen, T. Novak, O. Novgorodova, R. Novotny, L. Nozka, K. Ntekas, E. Nurse, F. Nuti,102 F. G. Oakham,33,e H. Oberlack,113 T. Obermann,24 J. Ocariz,132 A. Ochi,80 I. Ochoa,38 J. P. Ochoa-Ricoux,144a K. O’Connor,26 S. Oda,85 S. Odaka,79 S. Oerdek,51 A. Oh,98 S. H. Oh,47 C. C. Ohm,151 H. Oide,53b,53a H. Okawa,166 Y. Okazaki,83 Y. Okumura,160 T. Okuyama,79 A. Olariu,27b L. F. Oleiro Seabra,136a S. A. Olivares Pino,144a D. Oliveira Damazio,29 J. L. Oliver,1 M. J. R. Olsson,36 A. Olszewski,82 J. Olszowska,82 D. C. O’Neil,149 A. Onofre,136a,136e K. Onogi,115 P. U. E. Onyisi,11 H. Oppen,130 M. J. Oreglia,36 Y. Oren,158 D. Orestano,72a,72b E. C. Orgill,98 N. Orlando,61b A. A. O’Rourke,44 R. S. Orr,164 B. Osculati,53b,53a,a V. O’Shea,55 R. Ospanov,58a G. Otero y Garzon,30 H. Otono,85 M. Ouchrif,34d F. Ould-Saada,130 A. Ouraou,142 Q. Ouyang,15a M. Owen,55 R. E. Owen,21 V. E. Ozcan,12c N. Ozturk,8 y g J. Pacalt,126 H. A. Pacey,31 K. Pachal,149 A. Pacheco Pages,14 L. Pacheco Rodriguez,142 C. Padilla Aranda,14 S. Pagan Griso,18 M. Paganini,180 G. Palacino,63 S. Palazzo,40b,40a S. Palestini,35 M. Palka,81b D. Pallin,37 I. Panagoulias,10 C. E. Pandini,35 D. Paredes Hernandez,61b S. R. Paredes Saenz,131 B. Parida,58c A. J. Parker,87 K. A. Parker,44 M. A. Parker,31 F. Parodi,53b,53a J. A. Parsons,38 U. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Haddad,34e A. Hadef,58a S. Hageböck,24 M. Hagihara,166 H. Hakobyan,181,a M. Haleem,174 J. Haley,125 G. Halladjian,104 G. D. Hallewell,99 K. Hamacher,179 P. Hamal,126 K. Hamano,173 A. Hamilton,32a G. N. Hamity,146 024908-25 024908-25 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. V. P. Maleev,134 F. Malek,56 U. Mallik,75 D. Malon,6 C. Malone,31 S. Maltezos,10 S. Malyukov,35 J. Mamuzic,171 G. Mancini,49 I. Mandi´c,89 J. Maneira,136a L. Manhaes de Andrade Filho,78a J. Manjarres Ramos,46 K. H. Mankinen,94 A. Mann,112 A. Manousos,74 B. Mansoulie,142 J. D. Mansour,15a M. Mantoani,51 S. Manzoni,66a,66b G. Marceca,30 L. March,52 L. Marchese,131 G. Marchiori,132 M. Marcisovsky,137 C. A. Marin Tobon,35 M. Marjanovic,37 D. E. Marley,103 024908-26 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) F. Marroquim,78b Z. Marshall,18 M. U. F. Martensson,169 S. Marti-Garcia,171 C. B. Martin,122 T. A. Martin,175 V. J. Martin,48 B. Martin dit Latour,17 M. Martinez,14,w V. I. Martinez Outschoorn,100 S. Martin-Haugh,141 V. S. Martoiu,27b A. C. Martyniuk,92 A. Marzin,35 L. Masetti,97 T. Mashimo,160 R. Mashinistov,108 J. Masik,98 A. L. Maslennikov,120b,120a L. H. Mason,102 L. Massa,71a,71b P. Mastrandrea,5 A. Mastroberardino,40b,40a T. Masubuchi,160 P. Mättig,179 J. Maurer,27b B. Maˇcek,89 S. J. Maxﬁeld,88 D. A. Maximov,120b,120a R. Mazini,155 I. Maznas,159 S. M. Mazza,143 N. C. Mc Fadden,116 G. Mc Goldrick,164 S. P. Mc Kee,103 A. McCarn,103 T. G. McCarthy,113 L. I. McClymont,92 E. F. McDonald,102 J. A. Mcfayden,35 G. Mchedlidze,51 M. A. McKay,41 K. D. McLean,173 S. J. McMahon,141 P. C. McNamara,102 C. J. McNicol,175 R. A. McPherson,173,n J. E. Mdhluli,32c Z. A. Meadows,100 S. Meehan,145 T. Megy,50 S. Mehlhase,112 A. Mehta,88 T. Meideck,56 B. Meirose,42 D. Melini,171,ai B. R. Mellado Garcia,32c J. D. Mellenthin,51 M. Melo,28a F. Meloni,44 A. Melzer,24 S. B. Menary,98 E. D. Mendes Gouveia,136a L. Meng,88 X. T. Meng,103 A. Mengarelli,23b,23a S. Menke,113 E. Meoni,40b,40a S. Mergelmeyer,19 C. Merlassino,20 P. Mermod,52 L. Merola,67a,67b C. Meroni,66a F. S. Merritt,36 A. Messina,70a,70b J. Metcalfe,6 A. S. Mete,168 C. Meyer,133 J. Meyer,157 J-P. Meyer,142 H. Meyer Zu Theenhausen,59a F. Miano,153 R. P. Middleton,141 L. Mijovi´c,48 G. Mikenberg,177 M. Mikestikova,137 M. Mikuž,89 M. Milesi,102 A. Milic,164 D. A. Millar,90 D. W. Miller,36 A. Milov,177 D. A. Milstead,43a,43b A. A. Minaenko,140 M. Miñano Moya,171 I. A. Minashvili,156b A. I. Mincer,121 B. Mindur,81a M. Mineev,77 Y. Minegishi,160 Y. Ming,178 L. M. Mir,14 A. Mirto,65a,65b K. P. Mistry,133 T. Mitani,176 J. Mitrevski,112 V. A. Mitsou,171 A. Miucci,20 P. S. Miyagawa,146 A. Mizukami,79 J. U. Mjörnmark,94 T. Mkrtchyan,181 M. Mlynarikova,139 T. Moa,43a,43b K. Mochizuki,107 P. Mogg,50 S. Mohapatra,38 S. Molander,43a,43b R. Moles-Valls,24 M. C. Mondragon,104 K. Mönig,44 J. Monk,39 E. Monnier,99 A. Montalbano,149 J. Montejo Berlingen,35 F. Monticelli,86 S. Monzani,66a R. W. Moore,3 N. Morange,128 D. Moreno,22 M. Moreno Llácer,35 P. Morettini,53b M. Morgenstern,118 S. Morgenstern,46 D. Mori,149 T. Mori,160 M. Morii,57 M. Morinaga,176 V. Morisbak,130 A. K. Morley,35 G. Mornacchi,35 A. P. Morris,92 J. D. Morris,90 L. Morvaj,152 P. Moschovakos,10 M. Mosidze,156b H. J. Moss,146 J. Moss,150,aj K. Motohashi,162 R. Mount,150 E. Mountricha,35 E. J. W. Moyse,100 S. Muanza,99 F. Mueller,113 J. Mueller,135 R. S. P. Mueller,112 D. Muenstermann,87 P. Mullen,55 G. A. Mullier,20 F. J. Munoz Sanchez,98 P. Murin,28b W. J. Murray,175,141 A. Murrone,66a,66b M. Muškinja,89 C. Mwewa,32a A. G. Myagkov,140,ak J. Myers,127 M. Myska,138 B. P. Nachman,18 O. Nackenhorst,45 K. Nagai,131 K. Nagano,79 Y. Nagasaka,60 K. Nagata,166 M. Nagel,50 E. Nagy,99 A. M. Nairz,35 Y. Nakahama,115 K. Nakamura,79 T. Nakamura,160 I. Nakano,123 H. Nanjo,129 F. Napolitano,59a R. F. Naranjo Garcia,44 R. Narayan,11 D. I. Narrias Villar,59a I. Naryshkin,134 T. Naumann,44 G. Navarro,22 R. Nayyar,7 H. A. Neal,103 P. Y. Nechaeva,108 T. J. Neep,142 A. Negri,68a,68b M. Negrini,23b S. Nektarijevic,117 C. Nellist,51 M. E. Nelson,131 S. Nemecek,137 P. Nemethy,121 M. Nessi,35,al M. S. Neubauer,170 M. Neumann,179 P. R. Newman,21 T. Y. Ng,61c Y. S. Ng,19 H. D. N. Nguyen,99 T. Nguyen Manh,107 E. Nibigira,37 R. B. Nickerson,131 R. Nicolaidou,142 J. Nielsen,143 N. Nikiforou,11 V. Nikolaenko,140,ak I. Nikolic-Audit,132 K. Nikolopoulos,21 P. Nilsson,29 Y. Ninomiya,79 A. Nisati,70a N. Nishu,58c R. Nisius,113 I. Nitsche,45 T. Nitta,176 T. Nobe,160 Y. Noguchi,83 M. Nomachi,129 I. Nomidis,132 M. A. Nomura,29 T. Nooney,90 M. Nordberg,35 N. Norjoharuddeen,131 T. Novak,89 O. Novgorodova,46 R. Novotny,138 L. Nozka,126 K. Ntekas,168 E. Nurse,92 F. Nuti,102 F. G. Oakham,33,e H. Oberlack,113 T. Obermann,24 J. Ocariz,132 A. Ochi,80 I. Ochoa,38 J. P. Ochoa-Ricoux,144a K. O’Connor,26 S. Oda,85 S. Odaka,79 S. Oerdek,51 A. Oh,98 S. H. Oh,47 C. C. Ohm,151 H. Oide,53b,53a H. Okawa,166 Y. Okazaki,83 Y. Okumura,160 T. Okuyama,79 A. Olariu,27b L. F. Oleiro Seabra,136a S. A. Olivares Pino,144a D. Oliveira Damazio,29 J. L. Oliver,1 M. J. R. Olsson,36 A. Olszewski,82 J. Olszowska,82 D. C. O’Neil,149 A. Onofre,136a,136e K. Onogi,115 P. U. E. Onyisi,11 H. Oppen,130 M. J. Oreglia,36 Y. Oren,158 D. Orestano,72a,72b E. C. Orgill,98 N. Orlando,61b A. A. O’Rourke,44 R. S. Orr,164 B. Osculati,53b,53a,a V. O’Shea,55 R. Ospanov,58a G. Otero y Garzon,30 H. Otono,85 M. Ouchrif,34d F. Ould-Saada,130 A. Ouraou,142 Q. Ouyang,15a M. Owen,55 R. E. Owen,21 V. E. Ozcan,12c N. Ozturk,8 J. Pacalt,126 H. A. Pacey,31 K. Pachal,149 A. Pacheco Pages,14 L. Pacheco Rodriguez,142 C. Padilla Aranda,14 S. Pagan Griso,18 M. Paganini,180 G. Palacino,63 S. Palazzo,40b,40a S. Palestini,35 M. Palka,81b D. Pallin,37 I. Panagoulias,10 C. E. Pandini,35 J. G. Panduro Vazquez,91 P. Pani,35 G. Panizzo,64a,64c L. Paolozzi,52 T. D. Papadopoulou,10 K. Papageorgiou,9,t A. Paramonov,6 D. Paredes Hernandez,61b S. R. Paredes Saenz,131 B. Parida,58c A. J. Parker,87 K. A. Parker,44 M. A. Parker,31 F. Parodi,53b,53a J. A. Parsons,38 U. Parzefall,50 V. R. Pascuzzi,164 J. M. P. Pasner,143 E. Pasqualucci,70a S. Passaggio,53b F. Pastore,91 P. Pasuwan,43a,43b S. Pataraia,97 J. R. Pater,98 A. Pathak,178,f T. Pauly,35 B. Pearson,113 M. Pedersen,130 L. Pedraza Diaz,117 R. Pedro,136a,136b S. V. Peleganchuk,120b,120a O. Penc,137 C. Peng,15d H. Peng,58a B. S. Peralva,78a M. M. Perego,142 A. P. Pereira Peixoto,136a D. V. Perepelitsa,29 F. Peri,19 L. Perini,66a,66b H. Pernegger,35 S. Perrella,67a,67b V. D. Peshekhonov,77,a K. Peters,44 R. F. Y. Peters,98 B. A. Petersen,35 T. C. Petersen,39 E. Petit,56 A. Petridis,1 C. Petridou,159 P. Petroff,128 E. Petrolo,70a M. Petrov,131 F. Petrucci,72a,72b M. Pettee,180 N. E. Pettersson,100 A. Peyaud,142 R. Pezoa,144b T. Pham,102 F. H. Phillips,104 P. W. Phillips,141 G. Piacquadio,152 E. Pianori,18 A. Picazio,100 M. A. Pickering,131 R. Piegaia,30 36 98 71 71b 3 177 29 139 77 P. Rados,102 F. Ragusa,66a,66b G. Rahal,95 J. A. Raine,98 S. Rajagopalan,29 A. Ramirez Morales,90 T. Rashid,128 S. Raspopov,5 66 66b 44 112 9 146 1 98 35 M. G. Ratti,66a,66b D. M. Rauch,44 F. Rauscher,112 S. Rave,97 B. Ravina,146 I. Ravinovich,177 J. H. Rawling,98 M. Raymond,35 A. L. Read,130 N. P. Readioff,56 M. Reale,65a,65b D. M. Rebuzzi,68a,68b A. Redelbach,174 G. Redlinger,29 R. Reece,143 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Parzefall,50 V. R. Pascuzzi,164 J. M. P. Pasner,143 E. Pasqualucci,70a S. Passaggio,53b F. Pastore,91 P. Pasuwan,43a,43b S. Pataraia,97 J. R. Pater,98 A. Pathak,178,f T. Pauly,35 B. Pearson,113 M. Pedersen,130 L. Pedraza Diaz,117 R. Pedro,136a,136b S. V. Peleganchuk,120b,120a O. Penc,137 C. Peng,15d H. Peng,58a B. S. Peralva,78a M. M. Perego,142 136 29 19 66 66b 35 67 67b 77 A. P. Pereira Peixoto,136a D. V. Perepelitsa,29 F. Peri,19 L. Perini,66a,66b H. Pernegger,35 S. Perrella,67a,67b V. D. Peshekhonov,77,a K. Peters,44 R. F. Y. Peters,98 B. A. Petersen,35 T. C. Petersen,39 E. Petit,56 A. Petridis,1 C. Petridou,159 P. Petroff,128 E. Petrolo,70a M. Petrov,131 F. Petrucci,72a,72b M. Pettee,180 N. E. Pettersson,100 A. Peyaud,142 R. Pezoa,144b T. Pham,102 F. H. Phillips,104 P. W. Phillips,141 G. Piacquadio,152 E. Pianori,18 A. Picazio,100 M. A. Pickering,131 R. Piegaia,30 6 J. E. Pilcher,36 A. D. Pilkington,98 M. Pinamonti,71a,71b J. L. Pinfold,3 M. Pitt,177 M-A. Pleier,29 V. Pleskot,139 E. Plotnikova,77 D. Pluth,76 P. Podberezko,120b,120a R. Poettgen,94 R. Poggi,52 L. Poggioli,128 I. Pogrebnyak,104 D. Pohl,24 I. Pokharel,51 G. Polesello,68a A. Poley,44 A. Policicchio,40b,40a R. Polifka,35 A. Polini,23b C. S. Pollard,44 V. Polychronakos,29 D Ponomarenko 110 L Pontecorvo 70a G A Popeneciu 27d D M Portillo Quintero 132 S Pospisil 138 K Potamianos 44 J. E. Pilcher,36 A. D. Pilkington,98 M. Pinamonti,71a,71b J. L. Pinfold,3 M. Pitt,177 M-A. Pleier,29 V. Pleskot,139 E. Plotnikova,77 D. Pluth,76 P. Podberezko,120b,120a R. Poettgen,94 R. Poggi,52 L. Poggioli,128 I. Pogrebnyak,104 D. Pohl,24 I. Pokharel,51 G. Polesello,68a A. Poley,44 A. Policicchio,40b,40a R. Polifka,35 A. Polini,23b C. S. Pollard,44 V. Polychronakos,29 D. Ponomarenko,110 L. Pontecorvo,70a G. A. Popeneciu,27d D. M. Portillo Quintero,132 S. Pospisil,138 K. Potamianos,44 I. N. Potrap,77 C. J. Potter,31 H. Potti,11 T. Poulsen,94 J. Poveda,35 T. D. Powell,146 M. E. Pozo Astigarraga,35 P. Pralavorio,99 S. Prell,76 D. Price,98 M. Primavera,65a S. Prince,101 N. Proklova,110 K. Prokoﬁev,61c F. Prokoshin,144b S. Protopopescu,29 J. Proudfoot,6 M. Przybycien,81a A. Puri,170 P. Puzo,128 J. Qian,103 Y. Qin,98 A. Quadt,51 M. Queitsch-Maitland,44 A. Qureshi,1 024908-27 024908-27 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. p P. Rose,143 N-A. Rosien,51 E. Rossi,67a,67b L. P. Rossi,53b L. Rossini,66a,66b J. H. N. Rosten,31 R. Rosten,14 M. Rotaru,27b J. Rothberg,145 D. Rousseau,128 D. Roy,32c A. Rozanov,99 Y. Rozen,157 X. Ruan,32c F. Rubbo,150 F. Rühr,50 A. Ruiz-Martinez,171 50 77 101 7 44 134 170 Z. Rurikova,50 N. A. Rusakovich,77 H. L. Russell,101 J. P. Rutherfoord,7 E. M. Rüttinger,44,am Y. F. Ryabov,134 M. Rybar,170 G. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Rybkin,128 S. Ryu,6 A. Ryzhov,140 G. F. Rzehorz,51 P. Sabatini,51 G. Sabato,118 S. Sacerdoti,128 H. F-W. Sadrozinski,143 R. Sadykov,77 F. Safai Tehrani,70a P. Saha,119 M. Sahinsoy,59a A. Sahu,179 M. Saimpert,44 M. Saito,160 T. Saito,160 R. Sadykov, F. Safai Tehrani, P. Saha, M. Sahinsoy, A. Sahu, M. Saimpert, M. Saito, T. Saito, H. Sakamoto,160 A. Sakharov,121,af D. Salamani,52 G. Salamanna,72a,72b J. E. Salazar Loyola,144b D. Salek,118 P. H. Sales De Bruin,169 D. Salihagic,113 A. Salnikov,150 J. Salt,171 D. Salvatore,40b,40a F. Salvatore,153 A. Salvucci,61a,61b,61c A. Salzburger,35 35 50 159 159 1 1 64 64 130 g g J. Samarati,35 D. Sammel,50 D. Sampsonidis,159 D. Sampsonidou,159 J. Sánchez,171 A. Sanchez Pineda,64a,64c H. Sandaker,130 C O Sander 44 M Sandhoff 179 C Sandoval 22 D P C Sankey 141 M Sannino 53b,53a Y Sano 115 A Sansoni 49 C Santoni 37 y H. Santos,136a I. Santoyo Castillo,153 A. Sapronov,77 J. G. Saraiva,136a,136d O. Sasaki,79 K. Sato,166 E. Sauvan,5 P. Savard,164,e N. Savic,113 R. Sawada,160 C. Sawyer,141 L. Sawyer,93,v C. Sbarra,23b A. Sbrizzi,23b,23a T. Scanlon,92 J. Schaarschmidt,145 C. Schiavi,53b,53a S. Schier, 3 L. K. Schildgen, Z. M. Schillaci, 6 E. J. Schioppa,35 M. Schioppa, 0b, 0a K. E. Schleicher,50 S. Schlenker,35 K. R. Schmidt-Sommerfeld,113 K. Schmieden,35 C. Schmitt,97 S. Schmitt,44 S. Schmitz,97 U. Schnoor,50 L. Schoeffel,142 A. Schoening,59b E. Schopf,24 M. Schott,97 J. F. P. Schouwenberg,117 J. Schovancova,35 S. Schramm,52 A. Schulte,97 H-C. Schultz-Coulon,59a M. Schumacher,50 B. A. Schumm,143 Ph. Schune,142 A. Schwartzman,150 T. A. Schwarz,103 H. Schweiger,98 Ph. Schwemling,142 R. Schwienhorst,104 A. Sciandra,24 G. Sciolla,26 M. Scornajenghi,40b,40a F. Scuri,69a F. Scutti,102 L. M. Scyboz,113 J. Searcy,103 C. D. Sebastiani,70a,70b P. Seema,24 S. C. Seidel,116 A. Seiden,143 T. Seiss,36 J. M. Seixas,78b G. Sekhniaidze,67a K. Sekhon,103 S. J. Sekula,41 N. Semprini-Cesari,23b,23a S. Sen,47 S. Senkin,37 C. Serfon,130 L. Serin,128 L. Serkin,64a,64b M. Sessa,72a,72b H. Severini,124 F. Sforza,167 A. Sfyrla,52 E. Shabalina,51 y J. D. Shahinian,143 N. W. Shaikh,43a,43b L. Y. Shan,15a R. Shang,170 J. T. Shank,25 M. Shapiro,18 A. S. Sharma,1 A. Sharma,131 L. Shi,155,an S. Shimizu,80 C. O. Shimmin,180 M. Shimojima,114 I. P. J. Shipsey,131 S. Shirabe,85 M. Shiyakova,77 J. Shlomi,177 108 107 36 102 124 122 110 137 Shi,155,an S. Shimizu,80 C. O. Shimmin,180 M. Shi , , , , , , , , , R. Simoniello,97 P. Sinervo,164 N. B. Sinev,127 M. Sioli,23b,23a G. Siragusa,174 I. Siral,103 S. Yu. Sivoklokov,111 J. Sjölin,43a,43b M. B. Skinner,87 P. Skubic,124 M. Slater,21 T. Slavicek,138 M. J. Silva,136a M. Silva, Jr.,178 M. V. Silva Oliveira,78a S. B. Silverstein,43a L. Simic,77 S. Simion,128 E. Simioni,97 M. Simon,97 R. Simoniello,97 P. Sinervo,164 N. B. Sinev,127 M. Sioli,23b,23a G. Siragusa,174 I. Siral,103 S. Yu. Sivoklokov,111 J. Sjölin,43a,43b M. B. Skinner,87 P. Skubic,124 M. Slater,21 T. Slavicek,138 M. Slawinska,82 K. Sliwa,167 R. Slovak,139 V. Smakhtin,177 B. H. Smart,5 J. Smiesko,28a N. Smirnov,110 S. Yu. Smirnov,110 Y. Smirnov,110 L. N. Smirnova,111 O. Smirnova,94 J. W. Smith,51 M. N. K. Smith,38 R. W. Smith,38 M. Smizanska,87 K. Smolek,138 A. A. Snesarev,108 I. M. Snyder,127 S. Snyder,29 R. Sobie,173,n A. M. Soffa,168 A. Soffer,158 A. Søgaard,48 D. A. Soh,155 G. Sokhrannyi,89 C. A. Solans Sanchez,35 M. Solar,138 E. Yu. Soldatov,110 U. Soldevila,171 A. A. Solodkov,140 A. Soloshenko,77 O. V. Solovyanov,140 V. Solovyev,134 P. Sommer,146 H. Son,167 W. Song,141 A. Sopczak,138 F. Sopkova,28b D. Sosa,59b C. L. Sotiropoulou,69a,69b S. Sottocornola,68a,68b R. Soualah,64a,64c,ao A. M. Soukharev,120b,120a D. South,44 B. C. Sowden,91 S. Spagnolo,65a,65b M. Spalla,113 M. Spangenberg,175 F. Spanò,91 D. Sperlich,19 F. Spettel,113 T. M. Spieker,59a R. Spighi,23b G. Spigo,35 L. A. Spiller,102 D. P. Spiteri,55 M. Spousta,139 A. Stabile,66a,66b R. Stamen,59a S. Stamm,19 E. Stanecka,82 R. W. Stanek,6 C. Stanescu,72a B. Stanislaus,131 M. M. Stanitzki,44 B. Stapf,118 S. Stapnes,130 E. A. Starchenko,140 G. H. Stark,36 J. Stark,56 S. H Stark,39 P. Staroba,137 P. Starovoitov,59a S. Stärz,35 R. Staszewski,82 M. Stegler,44 P. Steinberg,29 B. Stelzer,149 H. J. Stelzer,35 O. Stelzer-Chilton,165a H. Stenzel,54 T. J. Stevenson,90 G. A. Stewart,55 M. C. Stockton,127 G. Stoicea,27b P. Stolte,51 S. Stonjek,113 A. Straessner,46 J. Strandberg,151 S. Strandberg,43a,43b M. Strauss,124 P. Strizenec,28b R. Ströhmer,174 D. M. Strom,127 R. Stroynowski,41 A. Strubig,48 S. A. Stucci,29 B. Stugu,17 J. Stupak,124 N. A. Styles,44 D. Su,150 J. Su,135 S. Suchek,59a Y. Sugaya,129 M. Suk,138 V. V. Sulin,108 D. M. S. Sultan,52 S. Sultansoy,4c T. Sumida,83 S. Sun,103 X. Sun,3 K. Suruliz,153 C. J. E. Suster,154 M. R. Sutton,153 S. Suzuki,79 M. Svatos,137 M. Swiatlowski,36 S. P. Swift,2 A. Sydorenko,97 I. Sykora,28a T. Sykora,139 D. Ta,97 K. Tackmann,44,ap J. Taenzer,158 A. Taffard,168 R. Taﬁrout,165a E. Tahirovic,90 N. Taiblum,158 H. Takai,29 R. Takashima,84 E. H. Takasugi,113 K. Takeda,80 T. Takeshita,147 Y. Takubo,79 M. Talby,99 A. A. Talyshev,120b,120a J. Tanaka,160 M. Tanaka,162 R. Tanaka,128 R. Tanioka,80 B. B. Tannenwald,122 S. Tapia Araya,144b S. Tapprogge,97 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Slawinska,82 K. Sliwa,167 R. Slovak,139 V. Smakhtin,177 B. H. Smart,5 J. Smiesko,28a N. Smirnov,110 S. Yu. Smirnov,110 Y. Smirnov,110 L. N. Smirnova,111 O. Smirnova,94 J. W. Smith,51 M. N. K. Smith,38 R. W. Smith,38 M. Smizanska,87 K. Smolek,138 A. A. Snesarev,108 I. M. Snyder,127 S. Snyder,29 R. Sobie,173,n A. M. Soffa,168 A. Soffer,158 A. Søgaard,48 D. A. Soh,155 G. Sokhrannyi,89 C. A. Solans Sanchez,35 M. Solar,138 E. Yu. Soldatov,110 U. Soldevila,171 A. A. Solodkov,140 A. Soloshenko,77 O. V. Solovyanov,140 V. Solovyev,134 P. Sommer,146 H. Son,167 W. Song,141 A. Sopczak,138 F. Sopkova,28b D. Sosa,59b C. L. Sotiropoulou,69a,69b S. Sottocornola,68a,68b R. Soualah,64a,64c,ao A. M. Soukharev,120b,120a D. South,44 B. C. Sowden,91 S. Spagnolo,65a,65b M. Spalla,113 M. Spangenberg,175 F. Spanò,91 D. Sperlich,19 F. Spettel,113 T. M. Spieker,59a R. Spighi,23b G. Spigo,35 L. A. Spiller,102 D. P. Spiteri,55 M. Spousta,139 A. Stabile,66a,66b R. Stamen,59a S. Stamm,19 E. Stanecka,82 R. W. Stanek,6 C. Stanescu,72a B. Stanislaus,131 M. M. Stanitzki,44 B. Stapf,118 S. Stapnes,130 E. A. Starchenko,140 G. H. Stark,36 J. Stark,56 S. H Stark,39 P. Staroba,137 P. Starovoitov,59a S. Stärz,35 R. Staszewski,82 M. Stegler,44 P. Steinberg,29 B. Stelzer,149 H. J. Stelzer,35 O. Stelzer-Chilton,165a H. Stenzel,54 T. J. Stevenson,90 G. A. Stewart,55 M. C. Stockton,127 G. Stoicea,27b P. Stolte,51 S. Stonjek,113 A. Straessner,46 J. Strandberg,151 S. Strandberg,43a,43b M. Strauss,124 P. Strizenec,28b R. Ströhmer,174 D. M. Strom,127 R. Stroynowski,41 A. Strubig,48 S. A. Stucci,29 B. Stugu,17 J. Stupak,124 N. A. Styles,44 D. Su,150 J. Su,135 S. Suchek,59a Y. Sugaya,129 M. Suk,138 V. V. Sulin,108 D. M. S. Sultan,52 S. Sultansoy,4c T. Sumida,83 S. Sun,103 X. Sun,3 K. Suruliz,153 C. J. E. Suster,154 M. R. Sutton,153 S. Suzuki,79 M. Svatos,137 M. Swiatlowski,36 S. P. Swift,2 A. Sydorenko,97 I. Sykora,28a T. Sykora,139 D. Ta,97 K. Tackmann,44,ap J. Taenzer,158 A. Taffard,168 R. Taﬁrout,165a E. Tahirovic,90 N. Taiblum,158 H. Takai,29 R. Takashima,84 E. H. Takasugi,113 K. Takeda,80 T. Takeshita,147 Y. Takubo,79 M. Talby,99 A. A. Talyshev,120b,120a J. Tanaka,160 M. Tanaka,162 R. Tanaka,128 R. Tanioka,80 B. B. Tannenwald,122 S. Tapia Araya,144b S. Tapprogge,97 024908-28 024908-28 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … 1Department of Physics, University of Adelaide, Adelaide, Australia 2Physics Department, SUNY Albany, Albany, New York, USA A. Tarek Abouelfadl Mohamed,132 S. Tarem,157 G. Tarna,27b,q G. F. Tartarelli,66a P. Tas,139 M. Tasevsky,137 T. Tashiro,83 E. Tassi,40b,40a A. Tavares Delgado,136a,136b Y. Tayalati,34e A. C. Taylor,116 A. J. Taylor,48 G. N. Taylor,102 P. T. E. Taylor,102 W. Taylor,165b A. S. Tee,87 P. Teixeira-Dias,91 H. Ten Kate,35 P. K. Teng,155 J. J. Teoh,118 F. Tepel,179 S. Terada,79 K. Terashi,160 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Urrejola,97 G. Usai,8 J. Usui,79 L. Vacavant,99 V. Vacek,138 B. Vachon,101 K. O. H. Vadla,130 A. Vaidya,92 C. Valderanis,112 E. Valdes Santurio,43a,43b M. Valente,52 S Valentinetti 23b,23a A Valero 171 L Valéry 44 R A Vallance 21 A Vallier 5 J A Valls Ferrer 171 T R Van Daalen 14 G. Unel,168 F. C. Ungaro,102 Y. Unno,79 K. Uno,160 J. Urban,28b P. Urquijo,102 P. Urrejola,97 G. Usai,8 J. Usui,79 L. Vacavant,99 V. Vacek,138 B. Vachon,101 K. O. H. Vadla,130 A. Vaidya,92 C. Valderanis,112 E. Valdes Santurio,43a,43b M. Valente,52 S. Valentinetti,23b,23a A. Valero,171 L. Valéry,44 R. A. Vallance,21 A. Vallier,5 J. A. Valls Ferrer,171 T. R. Van Daalen,14 y W. Van Den Wollenberg,118 H. Van der Graaf,118 P. Van Gemmeren,6 J. Van Nieuwkoop,149 I. Van Vulpen,118 M. Vanadia,71a,71b W. Vandelli,35 A. Vaniachine,163 P. Vankov,118 R. Vari,70a E. W. Varnes,7 C. Varni,53b,53a T. Varol,41 D. Varouchas,128 , , , , , , , , K. E. Varvell,154 G. A. Vasquez,144b J. G. Vasquez,180 F. Vazeille,37 D. Vazquez Furelos,14 T. Vazquez Schroeder,101 J. Veatch,51 V. Vecchio,72a,72b L. M. Veloce,164 F. Veloso,136a,136c S. Veneziano,70a A. Ventura,65a,65b M. Venturi,173 N. Venturi,35 V. Vercesi,68a M. Verducci,72a,72b C. M. Vergel Infante,76 W. Verkerke,118 A. T. Vermeulen,118 J. C. Vermeulen,118 M. C. Vetterli,149,e N. Viaux Maira,144b M. Vicente Barreto Pinto,52 I. Vichou,170,* T. Vickey,146 O. E. Vickey Boeriu,146 G. H. A. Viehhauser,131 S. Viel,18 L. Vigani,131 M. Villa,23b,23a M. Villaplana Perez,66a,66b E. Vilucchi,49 M. G. Vincter,33 V. B. Vinogradov,77 A. Vishwakarma,44 C. Vittori,23b,23a I. Vivarelli,153 S. Vlachos,10 M. Vogel,179 P. Vokac,138 G. Volpi,14 S. E. von Buddenbrock,32c E. Von Toerne,24 V. Vorobel,139 K. Vorobev,110 M. Vos,171 J. H. Vossebeld,88 N. Vranjes,16 16 138 118 Š 89 35 36 Ž 28 Ž 16 , , , , , , j , M. Vranjes Milosavljevic,16 V. Vrba,138 M. Vreeswijk,118 T. Šﬁligoj,89 R. Vuillermet,35 I. Vukotic,36 T. Ženiš,28a L. Živkovi´c,16 P. Wagner,24 W. Wagner,179 J. Wagner-Kuhr,112 H. Wahlberg,86 S. Wahrmund,46 K. Wakamiya,80 V. M. Walbrecht,113 J. Walder,87 R. Walker,112 W. Walkowiak,148 V. Wallangen,43a,43b A. M. Wang,57 C. Wang,58b,q F. Wang,178 H. Wang,18 H. Wang,3 J. Wang,154 J. Wang,59b P. Wang,41 Q. Wang,124 R.-J. Wang,132 R. Wang,58a R. Wang,6 S. M. Wang,155 W. T. Wang,58a W. Wang,155,at W. X. Wang,58a,au Y. Wang,58a Z. Wang,58c C. Wanotayaroj,44 A. Warburton,101 C. P. Ward,31 D. R. Wardrope,92 A. Washbrook,48 P. M. Watkins,21 A. T. Watson,21 M. F. Watson,21 G. Watts,145 S. Watts,98 B. M. A. Tarek Abouelfadl Mohamed,132 S. Tarem,157 G. Tarna,27b,q G. F. Tartarelli,66a P. Tas,139 M. Tasevsky,137 T. Tashiro,83 E. Tassi,40b,40a A. Tavares Delgado,136a,136b Y. Tayalati,34e A. C. Taylor,116 A. J. Taylor,48 G. N. Taylor,102 P. T. E. Taylor,102 W. Taylor,165b A. S. Tee,87 P. Teixeira-Dias,91 H. Ten Kate,35 P. K. Teng,155 J. J. Teoh,118 F. Tepel,179 S. Terada,79 K. Terashi,160 J. Terron,96 S. Terzo,14 M. Testa,49 R. J. Teuscher,164,n S. J. Thais,180 T. Theveneaux-Pelzer,44 F. Thiele,39 J. P. Thomas,21 A. S. Thompson,55 P. D. Thompson,21 L. A. Thomsen,180 E. Thomson,133 Y. Tian,38 R. E. Ticse Torres,51 V O Tikh i 108 aq Y A Tikh 120b 120a S Ti h k 110 P Ti t 180 S Ti t 99 K T d 162 MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … PHYSICAL REVIEW C 98, 024908 (2018) A. Tarek Abouelfadl Mohamed,132 S. Tarem,157 G. Tarna,27b,q G. F. Tartarelli,66a P. Tas,139 M. Tasevsky,137 T. Tashiro,83 E. Tassi,40b,40a A. Tavares Delgado,136a,136b Y. Tayalati,34e A. C. Taylor,116 A. J. Taylor,48 G. N. Taylor,102 P. T. E. Taylor,102 W. Taylor,165b A. S. Tee,87 P. Teixeira-Dias,91 H. Ten Kate,35 P. K. Teng,155 J. J. Teoh,118 F. Tepel,179 S. Terada,79 K. Terashi,160 J. Terron,96 S. Terzo,14 M. Testa,49 R. J. Teuscher,164,n S. J. Thais,180 T. Theveneaux-Pelzer,44 F. Thiele,39 J. P. Thomas,21 A. S. Thompson,55 P. D. Thompson,21 L. A. Thomsen,180 E. Thomson,133 Y. Tian,38 R. E. Ticse Torres,51 V. O. Tikhomirov,108,aq Yu. A. Tikhonov,120b,120a S. Timoshenko,110 P. Tipton,180 S. Tisserant,99 K. Todome,162 S. Todorova-Nova,5 S. Todt,46 J. Tojo,85 S. Tokár,28a K. Tokushuku,79 E. Tolley,122 K. G. Tomiwa,32c M. Tomoto,115 L. Tompkins,150 K. Toms,116 B. Tong,57 P. Tornambe,50 E. Torrence,127 H. Torres,46 E. Torró Pastor,145 C. Tosciri,131 J. Toth,99,ar F. Touchard,99 D. R. Tovey,146 C. J. Treado,121 T. Trefzger,174 F. Tresoldi,153 A. Tricoli,29 I. M. Trigger,165a S. Trincaz-Duvoid,132 M. F. Tripiana,14 W. Trischuk,164 B. Trocmé,56 A. Trofymov,128 C. Troncon,66a M. Trovatelli,173 S. Trincaz-Duvoid,132 M. F. Tripiana,14 W. Trischuk,164 B. Trocmé,56 A. Trofymov,128 C. Troncon,66a M. Trovatelli,173 F. Trovato,153 L. Truong,32b M. Trzebinski,82 A. Trzupek,82 F. Tsai,44 J. C.-L. Tseng,131 P. V. Tsiareshka,105 N. Tsirintanis,9 V. Tsiskaridze,152 E. G. Tskhadadze,156a I. I. Tsukerman,109 V. Tsulaia,18 S. Tsuno,79 D. Tsybychev,152 Y. Tu,61b A. Tudorache,27b V. Tudorache,27b T. T. Tulbure,27a A. N. Tuna,57 S. Turchikhin,77 D. Turgeman,177 I. Turk Cakir,4b,as 66 38 97 23b 23 79 43 43b 166 35 29 p y F. Trovato,153 L. Truong,32b M. Trzebinski,82 A. Trzupek,82 F. Tsai,44 J. C.-L. Tseng,131 P. V. Tsiareshka,105 N. Tsirintanis,9 V. Tsiskaridze,152 E. G. Tskhadadze,156a I. I. Tsukerman,109 V. Tsulaia,18 S. Tsuno,79 D. Tsybychev,152 Y. Tu,61b A. Tudorache,27b V. Tudorache,27b T. T. Tulbure,27a A. N. Tuna,57 S. Turchikhin,77 D. Turgeman,177 I. Turk Cakir,4b,as g R. Turra,66a P. M. Tuts,38 E. Tzovara,97 G. Ucchielli,23b,23a I. Ueda,79 M. Ughetto,43a,43b F. Ukeg 168 102 79 160 28b 102 97 g R. Turra,66a P. M. Tuts,38 E. Tzovara,97 G. Ucchielli,23b,23a I. Ueda,79 M. Ughetto,43a,43b F. Ukegawa,166 G. Unal,35 A. Undrus,29 168 102 79 160 28b 102 97 8 79 99 R. Turra,66a P. M. Tuts,38 E. Tzovara,97 G. Ucchielli,23b,23a I. Ueda,79 M. Ughetto,43a,43b F. Ukegawa,166 G. Unal,35 A. Undrus,29 G. Unel,168 F. C. Ungaro,102 Y. Unno,79 K. Uno,160 J. Urban,28b P. Urquijo,102 P. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Waugh,92 A. F. Webb,11 S. Webb,97 C. Weber,180 M. S. Weber,20 S. A. Weber,33 S. M. Weber,59a J. S. Webster,6 A. R. Weidberg,131 g g g M. D. Werner,76 P. Werner,35 M. Wessels,59a T. D. Weston,20 K. Whalen,127 N. L. Whallon,145 A. M. Wharton,87 A. S. White,103 g S. Williams,31 C. Willis,104 S. Willocq,100 J. A. Wilson,21 I. Wingerter-Seez,5 E. Winkels,153 F. Winklmeier,127 O. J. Winston,153 B. T. Winter,24 M. Wittgen,150 M. Wobisch,93 A. Wolf,97 T. M. H. Wolf,118 R. Wolff,99 M. W. Wolter,82 H. Wolters,136a,136c V. W. S. Wong,172 N. L. Woods,143 S. D. Worm,21 B. K. Wosiek,82 K. W. Wo´zniak,82 K. Wraight,55 M. Wu,36 S. L. Wu,178 X. Wu,52 Y. Wu,58a T. R. Wyatt,98 B. M. Wynne,48 S. Xella,39 Z. Xi,103 L. Xia,175 D. Xu,15a H. Xu,58a L. Xu,29 T. Xu,142 W. Xu,103 B. Yabsley,154 S. Yacoob,32a K. Yajima,129 D. P. Yallup,92 D. Yamaguchi,162 Y. Yamaguchi,162 A. Yamamoto,79 T. Yamanaka,160 F. Yamane,80 M. Yamatani,160 T. Yamazaki,160 Y. Yamazaki,80 Z. Yan,25 H. J. Yang,58c,58d H. T. Yang,18 S. Yang,75 Y. Yang,160 Z. Yang,17 W-M. Yao,18 Y. C. Yap,44 Y. Yasu,79 E. Yatsenko,58c,58d J. Ye,41 S. Ye,29 I. Yeletskikh,77 E. Yigitbasi,25 E. Yildirim,97 K. Yorita,176 K. Yoshihara,133 C. J. S. Young,35 C. Young,150 J. Yu,8 J. Yu,76 X. Yue,59a 24 82 10 52 14 140 k 44 17 g g g S. P. Y. Yuen,24 B. Zabinski,82 G. Zacharis,10 E. Zaffaroni,52 R. Zaidan,14 A. M. Zaitsev,140,ak N. Zakharchuk,44 J. Zalieckas,17 S. Zambito,57 D. Zanzi,35 D. R. Zaripovas,55 S. V. Zeißner,45 C. Zeitnitz,179 G. Zemaityte,131 J. C. Zeng,170 Q. Zeng,150 O. Zenin,140 D. Zerwas,128 M. Zgubiˇc,131 D. F. Zhang,58b D. Zhang,103 F. Zhang,178 G. Zhang,58a H. Zhang,15c J. Zhang,6 L. Zhang,15c L. Zhang,58a M. Zhang,170 P. Zhang,15c R. Zhang,58a R. Zhang,24 X. Zhang,58b Y. Zhang,15d Z. Zhang,128 X. Zhao,41 Y. Zhao,58b,128,y Z. Zhao,58a A. Zhemchugov,77 B. Zhou,103 C. Zhou,178 L. Zhou,41 M. S. Zhou,15d M. Zhou,152 N. Zhou,58c Y. Zhou,7 C. G. Zhu,58b H. L. Zhu,58a H. Zhu,15a J. Zhu,103 Y. Zhu,58a X. Zhuang,15a K. Zhukov,108 V. Zhulanov,120b,120a A. Zibell,174 D. Zieminska,63 N. I. Zimine,77 S. Zimmermann,50 Z. Zinonos,113 M. Zinser,97 M. Ziolkowski,148 G. Zobernig,178 A. Zoccoli,23b,23a K. Zoch,51 T. G. Zorbas,146 R. Zou,36 M. Zur Nedden,19 and L. Zwalinski35 (ATLAS Collaboration) 024908-29 PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. M. AABOUD et al. (ATLAS Collaboration) PHYSICAL REVIEW C 98, 024908 (2018 3Department of Physics, University of Alberta, Edmonton, Alberta, Canada 4aDepartment of Physics, Ankara University, Ankara, Turkey 4bIstanbul Aydin University, Istanbul, Turkey 4cDivision of Physics, TOBB University of Economics and Technology, Ankara, Turkey 5LAPP, Université Grenoble Alpes, Université Savoie Mont Blanc, CNRS/IN2P3, Annecy, France 6High Energy Physics Division, Argonne National Laboratory, Argonne, Illinois, USA 7Department of Physics, University of Arizona, Tucson, Arizona, USA 8Department of Physics, University of Texas at Arlington, Arlington, Texas, USA 9Physics Department, National and Kapodistrian University of Athens, Athens, Greece 10Physics Department, National Technical University of Athens, Zografou, Greece 11Department of Physics, University of Texas at Austin, Austin, Texas, USA 12aBahcesehir University, Faculty of Engineering and Natural Sciences, Istanbul, Turkey 12bIstanbul Bilgi University, Faculty of Engineering and Natural Sciences, Istanbul, Turkey 12cDepartment of Physics, Bogazici University, Istanbul, Turkey 12dDepartment of Physics Engineering, Gaziantep University, Gaziantep, Turkey 13Institute of Physics, Azerbaijan Academy of Sciences, Baku, Azerbaijan 14Institut de Física d’Altes Energies (IFAE), Barcelona Institute of Science and Technology, Barcelona, Spain 15aInstitute of High Energy Physics, Chinese Academy of Sciences, Beijing, China 15bPhysics Department, Tsinghua University, Beijing, China 15cDepartment of Physics, Nanjing University, Nanjing, China 15dUniversity of Chinese Academy of Science (UCAS), Beijing, China 16Institute of Physics, University of Belgrade, Belgrade, Serbia 17Department for Physics and Technology, University of Bergen, Bergen, Norway 18Physics Division, Lawrence Berkeley National Laboratory and University of California, Berkeley, California, USA 19Institut für Physik, Humboldt Universität zu Berlin, Berlin, Germany 20Albert Einstein Center for Fundamental Physics and Laboratory for High Energy Physics, University of Bern, Bern, Switzerland 21School of Physics and Astronomy, University of Birmingham, Birmingham, United Kingdom 22Centro de Investigaciónes, Universidad Antonio Nariño, Bogota, Colombia 23aDipartimento di Fisica e Astronomia, Università di Bologna, Bologna, Italy 23bINFN Sezione di Bologna, Italy 24Physikalisches Institut, Universität Bonn, Bonn, Germany 25Department of Physics, Boston University, Boston, Massachusetts, USA 26Department of Physics, Brandeis University, Waltham, Massachusetts, USA 27aTransilvania University of Brasov, Brasov, Romania 27bHoria Hulubei National Institute of Physics and Nuclear Engineering, Bucharest, Romania 27cDepartment of Physics, Alexandru Ioan Cuza University of Iasi, Iasi, Romania 27dNational Institute for Research and Development of Isotopic and Molecular Technologies, Physics Department, Cluj-Napoca, Romania 27eUniversity Politehnica Bucharest, Bucharest, Romania 27fWest University in Timisoara, Timisoara, Romania 28aFaculty of Mathematics, Physics and Informatics, Comenius University, Bratislava 28bDepartment of Subnuclear Physics, Institute of Experimental Physics of the Slovak Academy of Sciences, Kosice, Slovak Republic 29Physics Department, Brookhaven National Laboratory, Upton, New York, USA 30Departamento de Física, Universidad de Buenos Aires, Buenos Aires, Argentina 31Cavendish Laboratory, University of Cambridge, Cambridge, United Kingdom 32aDepartment of Physics, University of Cape Town, Cape Town, South Africa 32bDepartment of Mechanical Engineering Science, University of Johannesburg, Johannesburg, Southo Africa 32cSchool of Physics, University of the Witwatersrand, Johannesburg, South Africa 33Department of Physics, Carleton University, Ottawa, Ontario, Canada 34aFaculté des Sciences Ain Chock, Réseau Universitaire de Physique des Hautes Energies - Université Hassan II, Casablanca, Morocco 34bCentre National de l’Energie des Sciences Techniques Nucleaires (CNESTEN), Rabat, Morocco 34cFaculté des Sciences Semlalia, Université Cadi Ayyad, LPHEA-Marrakech, Morocco 34dFaculté des Sciences, Université Mohamed Premier and LPTPM, Oujda, Morocco 34eFaculté des sciences, Université Mohammed V, Rabat, Morocco 35CERN, Geneva, Switzerland 36Enrico Fermi Institute, University of Chicago, Chicago, Illinois, USA 37LPC, Université Clermont Auvergne, CNRS/IN2P3, Clermont-Ferrand, France 38Nevis Laboratory, Columbia University, Irvington, New York, USA 39Niels Bohr Institute, University of Copenhagen, Copenhagen, Denmark 024908-30 3Department of Physics, University of Alberta, Edmonton, Alberta, Canada 4aDepartment of Physics, Ankara University, Ankara, Turkey 4bIstanbul Aydin University, Istanbul, Turkey 4cDivision of Physics, TOBB University of Economics and Technology, Ankara, Turkey 5LAPP, Université Grenoble Alpes, Université Savoie Mont Blanc, CNRS/IN2P3, Annecy, France 6High Energy Physics Division, Argonne National Laboratory, Argonne, Illinois, USA 7Department of Physics, University of Arizona, Tucson, Arizona, USA 8Department of Physics, University of Texas at Arlington, Arlington, Texas, USA 9Physics Department, National and Kapodistrian University of Athens, Athens, Greece 10Physics Department, National Technical University of Athens, Zografou, Greece 11Department of Physics, University of Texas at Austin, Austin, Texas, USA 12aBahcesehir University, Faculty of Engineering and Natural Sciences, Istanbul, Turkey 12bIstanbul Bilgi University, Faculty of Engineering and Natural Sciences, Istanbul, Turkey 12cDepartment of Physics, Bogazici University, Istanbul, Turkey 12dDepartment of Physics Engineering, Gaziantep University, Gaziantep, Turkey 13Institute of Physics, Azerbaijan Academy of Sciences, Baku, Azerbaijan 14Institut de Física d’Altes Energies (IFAE), Barcelona Institute of Science and Technology, Barcelona, Spain 15aInstitute of High Energy Physics, Chinese Academy of Sciences, Beijing, China 15bPhysics Department, Tsinghua University, Beijing, China 15cDepartment of Physics, Nanjing University, Nanjing, China 15dUniversity of Chinese Academy of Science (UCAS), Beijing, China 16Institute of Physics, University of Belgrade, Belgrade, Serbia 17Department for Physics and Technology, University of Bergen, Bergen, Norway 18Physics Division, Lawrence Berkeley National Laboratory and University of California, Berkeley, California, USA 19Institut für Physik, Humboldt Universität zu Berlin, Berlin, Germany 20Albert Einstein Center for Fundamental Physics and Laboratory for High Energy Physics, University of Bern, Bern, Switzerland 21School of Physics and Astronomy, University of Birmingham, Birmingham, United Kingdom 22Centro de Investigaciónes, Universidad Antonio Nariño, Bogota, Colombia 23aDipartimento di Fisica e Astronomia, Università di Bologna, Bologna, Italy 23bINFN Sezione di Bologna, Italy 24Physikalisches Institut, Universität Bonn, Bonn, Germany 25Department of Physics, Boston University, Boston, Massachusetts, USA 26Department of Physics, Brandeis University, Waltham, Massachusetts, USA 27aTransilvania University of Brasov, Brasov, Romania 27bHoria Hulubei National Institute of Physics and Nuclear Engineering, Bucharest, Romania 27cDepartment of Physics, Alexandru Ioan Cuza University of Iasi, Iasi, Romania 27dNational Institute for Research and Development of Isotopic and Molecular Technologies, Physics Department, Cluj-Napoca, Romania 27eUniversity Politehnica Bucharest, Bucharest, Romania 27fWest University in Timisoara, Timisoara, Romania 28aFaculty of Mathematics, Physics and Informatics, Comenius University, Bratislava 28bDepartment of Subnuclear Physics, Institute of Experimental Physics of the Slovak Academy of Sciences, Kosice, Slovak Republic 29Physics Department, Brookhaven National Laboratory, Upton, New York, USA 30Departamento de Física, Universidad de Buenos Aires, Buenos Aires, Argentina 31Cavendish Laboratory, University of Cambridge, Cambridge, United Kingdom 32aDepartment of Physics, University of Cape Town, Cape Town, South Africa 32bDepartment of Mechanical Engineering Science, University of Johannesburg, Johannesburg, Southo Africa 32cSchool of Physics, University of the Witwatersrand, Johannesburg, South Africa 33Department of Physics, Carleton University, Ottawa, Ontario, Canada 34aFaculté des Sciences Ain Chock, Réseau Universitaire de Physique des Hautes Energies - Université Hassan II, Casablanca, Morocco 34bCentre National de l’Energie des Sciences Techniques Nucleaires (CNESTEN), Rabat, Morocco 34cFaculté des Sciences Semlalia, Université Cadi Ayyad, LPHEA-Marrakech, Morocco 34dFaculté des Sciences, Université Mohamed Premier and LPTPM, Oujda, Morocco 34eFaculté des sciences, Université Mohammed V, Rabat, Morocco 35CERN, Geneva, Switzerland 36Enrico Fermi Institute, University of Chicago, Chicago, Illinois, USA 37LPC, Université Clermont Auvergne, CNRS/IN2P3, Clermont-Ferrand, France 38Nevis Laboratory, Columbia University, Irvington, New York, USA 39Niels Bohr Institute, University of Copenhagen, Copenhagen, Denmark 024908-30 PHYSICAL REVIEW C 98, 024908 (2018) MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … SU O J G O + S C C 98, 0 908 ( 0 40aDipartimento di Fisica, Università della Calabria, Rende 40bINFN Gruppo Collegato di Cosenza, Laboratori Nazionali di Frascati, Italy 41Physics Department, Southern Methodist University, Dallas, Texas, USA 42Physics Department, University of Texas at Dallas, Richardson, Texas, USA 43aDepartment of Physics, Stockholm University, Stockholm, Sweden 43bOskar Klein Centre, Stockholm, Sweden 44Deutsches Elektronen-Synchrotron DESY, Hamburg and Zeuthen, Germany 45Lehrstuhl für Experimentelle Physik IV, Technische Universität Dortmund, Dortmund, Germany 46Institut für Kern und Teilchenphysik, Technische Universität Dresden, Dresden, Germany 47Department of Physics, Duke University, Durham, North Carolina, USA 48SUPA - School of Physics and Astronomy, University of Edinburgh, Edinburgh, United Kingdom 49INFN e Laboratori Nazionali di Frascati, Frascati, Italy 50Physikalisches Institut, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany 51II. Physikalisches Institut, Georg-August-Universität Göttingen, Göttingen, Germany 52Département de Physique Nucléaire et Corpusculaire, Université de Genève, Genève, Switzerland 53aDipartimento di Fisica, Università di Genova, Genova 53bINFN Sezione di Genova, Italy 54II. PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. 76Department of Physics and Astronomy, Iowa State University, Ames, Iowa, USA 77Joint Institute for Nuclear Research, Dubna, Russia 78aDepartamento de Engenharia Elétrica, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, Brazil 78bUniversidade Federal do Rio De Janeiro COPPE/EE/IF, Rio de Janeiro, Brazil 78cUniversidade Federal de São João del Rei (UFSJ), São João del Rei, Brazil 78dInstituto de Física, Universidade de São Paulo, São Paulo, Brazil 79KEK, High Energy Accelerator Research Organization, Tsukuba, Japan 80Graduate School of Science, Kobe University, Kobe, Japan 81aAGH University of Science and Technology, Faculty of Physics and Applied Computer Science, Krakow 81bMarian Smoluchowski Institute of Physics, Jagiellonian University, Krakow, Poland 82Institute of Nuclear Physics Polish Academy of Sciences, Krakow, Poland 83Faculty of Science, Kyoto University, Kyoto, Japan 84Kyoto University of Education, Kyoto, Japan 85Research Center for Advanced Particle Physics and Department of Physics, Kyushu University, Fukuoka, Japan 86Instituto de Física La Plata, Universidad Nacional de La Plata and CONICET, La Plata, Argentina 87Physics Department, Lancaster University, Lancaster, United Kingdom 88Oliver Lodge Laboratory, University of Liverpool, Liverpool, United Kingdom 89Department of Experimental Particle Physics, Jožef Stefan Institute and Department of Physics, University of Ljubljana, Ljubljana, Slovenia 90School of Physics and Astronomy, Queen Mary University of London, London, United Kingdom 91Department of Physics, Royal Holloway University of London, Egham, United Kingdom 92Department of Physics and Astronomy, University College London, London, United Kingdom 93Louisiana Tech University, Ruston, Louisiana, USA 94Fysiska institutionen, Lunds Universitet, Lund, Sweden 95Centre de Calcul de l’Institut National de Physique Nucléaire et de Physique des Particules (IN2P3), Villeurbanne, France 96Departamento de Física Teorica C-15 and CIAFF, Universidad Autónoma de Madrid, Madrid, Spain 97Institut für Physik, Universität Mainz, Mainz, Germany 98School of Physics and Astronomy, University of Manchester, Manchester, United Kingdom 99CPPM, Aix-Marseille Université, CNRS/IN2P3, Marseille, France 100Department of Physics, University of Massachusetts, Amherst, Massachusetts, USA 101Department of Physics, McGill University, Montreal, Quebec, Canada 102School of Physics, University of Melbourne, Victoria, Australia 103Department of Physics, University of Michigan, Ann Arbor, Michigan, USA 104Department of Physics and Astronomy, Michigan State University, East Lansing, Michigan, USA 105B.I. Stepanov Institute of Physics, National Academy of Sciences of Belarus, Minsk, Belarus 106Research Institute for Nuclear Problems of Byelorussian State University, Minsk, Belarus 107Group of Particle Physics, University of Montreal, Montreal, Quebec, Canada 108P.N. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Physikalisches Institut, Justus-Liebig-Universität Giessen, Giessen, Germany 55SUPA - School of Physics and Astronomy, University of Glasgow, Glasgow, United Kingdom 56LPSC, Université Grenoble Alpes, CNRS/IN2P3, Grenoble INP, Grenoble, France 57Laboratory for Particle Physics and Cosmology, Harvard University, Cambridge, Massachusetts, USA 58aDepartment of Modern Physics and State Key Laboratory of Particle Detection and Electronics, University of Science and Technology of China, Hefei 58bInstitute of Frontier and Interdisciplinary Science and Key Laboratory of Particle Physics and Particle Irradiation (MOE), Shandong University, Qingdao 58cSchool of Physics and Astronomy, Shanghai Jiao Tong University, KLPPAC-MoE, SKLPPC, Shanghai 58dTsung-Dao Lee Institute, Shanghai, China 59aKirchhoff-Institut für Physik, Ruprecht-Karls-Universität Heidelberg, Heidelberg 59bPhysikalisches Institut, Ruprecht-Karls-Universität Heidelberg, Heidelberg, Germany 60Faculty of Applied Information Science, Hiroshima Institute of Technology, Hiroshima, Japan 61aDepartment of Physics, Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China 61bDepartment of Physics, University of Hong Kong, Hong Kong, China 61cDepartment of Physics and Institute for Advanced Study, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China 62Department of Physics, National Tsing Hua University, Hsinchu, Taiwan 63Department of Physics, Indiana University, Bloomington, Indiana, USA 64aINFN Gruppo Collegato di Udine, Sezione di Trieste, Udine, Italy 64bICTP, Trieste, Italy 64cDipartimento di Chimica, Fisica e Ambiente, Università di Udine, Udine, Italy 65aINFN Sezione di Lecce, Italy 65bDipartimento di Matematica e Fisica, Università del Salento, Lecce, Italy 66aINFN Sezione di Milano, Italy 66bDipartimento di Fisica, Università di Milano, Milano, Italy 67aINFN Sezione di Napoli, Italy 67bDipartimento di Fisica, Università di Napoli, Napoli, Italy 68aINFN Sezione di Pavia, Italy 68bDipartimento di Fisica, Università di Pavia, Pavia, Italy 69aINFN Sezione di Pisa, Italy 69bDipartimento di Fisica E. Fermi, Università di Pisa, Pisa, Italy 70aINFN Sezione di Roma, Italy 70bDipartimento di Fisica, Sapienza Università di Roma, Roma, Italy 71aINFN Sezione di Roma Tor Vergata, Italy 71bDipartimento di Fisica, Università di Roma Tor Vergata, Roma, Italy 72aINFN Sezione di Roma Tre, Italy 72bDipartimento di Matematica e Fisica, Università Roma Tre, Roma, Italy 73aINFN-TIFPA, Italy 73bUniversità degli Studi di Trento, Trento, Italy 74Institut für Astro und Teilchenphysik, Leopold-Franzens-Universität, Innsbruck, Austria 75University of Iowa, Iowa City, Iowa, USA 024908-31 PHYSICAL REVIEW C 98, 024908 (2018) PHYSICAL REVIEW C 98, 024908 (2018) Lebedev Physical Institute of the Russian Academy of Sciences, Moscow, Russia 109Institute for Theoretical and Experimental Physics (ITEP), Moscow, Russia 110National Research Nuclear University MEPhI, Moscow, Russia 111D.V. Skobeltsyn Institute of Nuclear Physics, M.V. Lomonosov Moscow State University, Moscow, Russia 112Fakultät für Physik, Ludwig-Maximilians-Universität München, München, Germany 113Max-Planck-Institut für Physik (Werner-Heisenberg-Institut), München, Germany 114Nagasaki Institute of Applied Science, Nagasaki, Japan 115Graduate School of Science and Kobayashi-Maskawa Institute, Nagoya University, Nagoya, Japan 116Department of Physics and Astronomy, University of New Mexico, Albuquerque, New Mexico, USA 117Institute for Mathematics, Astrophysics and Particle Physics, Radboud University Nijmegen/Nikhef, Nijmegen, Netherlands 118Nikhef National Institute for Subatomic Physics and University of Amsterdam, Amsterdam, Netherlands 119Department of Physics, Northern Illinois University, DeKalb, Illinois, USA 120aBudker Institute of Nuclear Physics, SB RAS, Novosibirsk 120bNovosibirsk State University Novosibirsk, Russia 121Department of Physics, New York University, New York, New York, USA 122Ohio State University, Columbus, Ohio, USA 123Faculty of Science, Okayama University, Okayama, Japan 124Homer L. Dodge Department of Physics and Astronomy, University of Oklahoma, Norman, Oklahoma, USA 125Department of Physics, Oklahoma State University, Stillwater, Oklahoma, USA 126Palacký University, RCPTM, Joint Laboratory of Optics, Olomouc, Czech Republic 127Center for High Energy Physics, University of Oregon, Eugene, Oregon, USA 128LAL, Université Paris-Sud, CNRS/IN2P3, Université Paris-Saclay, Orsay, France 024908-32 PHYSICAL REVIEW C 98, 024908 (2018) MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … 129Graduate School of Science, Osaka University, Osaka, Japan 130Department of Physics, University of Oslo, Oslo, Norway 131Department of Physics, Oxford University, Oxford, United Kingdom 132LPNHE, Sorbonne Université, Paris Diderot Sorbonne Paris Cité, CNRS/IN2P3, Paris, France 133Department of Physics, University of Pennsylvania, Philadelphia, Pennsylvania, USA 134Konstantinov Nuclear Physics Institute of National Research Centre “Kurchatov Institute,” PNPI, St. Petersburg, Russia 135Department of Physics and Astronomy, University of Pittsburgh, Pittsburgh, Pennsylvania, USA 136aLaboratório de Instrumentação e Física Experimental de Partículas - LIP, Lisboa, Portugal 136bDepartamento de Física, Faculdade de Ciências, Universidade de Lisboa, Lisboa, Portugal 136cDepartamento de Física, Universidade de Coimbra, Coimbra, Portugal 136dCentro de Física Nuclear da Universidade de Lisboa, Lisboa, Portugal 136eDepartamento de Física, Universidade do Minho, Braga, Portugal 136fDepartamento de Física Teorica y del Cosmos, Universidad de Granada, Granada, Spain 136gDep Física and CEFITEC of Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal 137Institute of Physics, Academy of Sciences of the Czech Republic, Prague,Czech Republic 138Czech Technical University in Prague, Prague, Czech Republic 139Charles University, Faculty of Mathematics and Physics, Prague, Czech Republic 140State Research Center Institute for High Energy Physics, NRC KI, Protvino, Russia 141Particle Physics Department, Rutherford Appleton Laboratory, Didcot, United Kingdom 142IRFU, CEA, Université Paris-Saclay, Gif-sur-Yvette, France 143Santa Cruz Institute for Particle Physics, University of California Santa Cruz, Santa Cruz, California, USA 144aDepartamento de Física, Pontiﬁcia Universidad Católica de Chile, Santiago, Chile 144bDepartamento de Física, Universidad Técnica Federico Santa María, Valparaíso, Chile 145Department of Physics, University of Washington, Seattle, Washington, USA 146Department of Physics and Astronomy, University of Shefﬁeld, Shefﬁeld, United Kingdom 147Department of Physics, Shinshu University, Nagano, Japan 148Department Physik, Universität Siegen, Siegen, Germany 149Department of Physics, Simon Fraser University, Burnaby, British Columbia, Canada 150SLAC National Accelerator Laboratory, Stanford, California, USA 151Physics Department, Royal Institute of Technology, Stockholm, Sweden 152Departments of Physics and Astronomy, Stony Brook University, Stony Brook, New York, USA 153Department of Physics and Astronomy, University of Sussex, Brighton, United Kingdom 154School of Physics, University of Sydney, Sydney, Australia 155Institute of Physics, Academia Sinica, Taipei, Taiwan 156aE. Andronikashvili Institute of Physics, Iv. MEASUREMENT OF JET FRAGMENTATION IN Pb+Pb AND … Javakhishvili Tbilisi State University, Tbilisi, Georgia 156bHigh Energy Physics Institute, Tbilisi State University, Tbilisi, Georgia 157Department of Physics, Technion, Israel Institute of Technology, Haifa, Israel 158Raymond and Beverly Sackler School of Physics and Astronomy, Tel Aviv University, Tel Aviv, Israel 159Department of Physics, Aristotle University of Thessaloniki, Thessaloniki, Greece 160International Center for Elementary Particle Physics and Department of Physics, University of Tokyo, Tokyo, Japan 161Graduate School of Science and Technology, Tokyo Metropolitan University, Tokyo, Japan 162Department of Physics, Tokyo Institute of Technology, Tokyo, Japan 163Tomsk State University, Tomsk, Russia 164Department of Physics, University of Toronto, Toronto, Ontario, Canada 165aTRIUMF, Vancouver BC 165bDepartment of Physics and Astronomy, York University, Toronto, Ontario, Canada 166Division of Physics and Tomonaga Center for the History of the Universe, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba Japan 167Department of Physics and Astronomy, Tufts University, Medford, Massachusetts, USA 168Department of Physics and Astronomy, University of California Irvine, Irvine, California, USA 169Department of Physics and Astronomy, University of Uppsala, Uppsala, Sweden 170Department of Physics, University of Illinois, Urbana, Illinois, USA 171Instituto de Física Corpuscular (IFIC), Centro Mixto Universidad de Valencia - CSIC, Valencia, Spain 172Department of Physics, University of British Columbia, Vancouver, British Columbia, Canada 173Department of Physics and Astronomy, University of Victoria, Victoria, British Columbia, Canada 174Fakultät für Physik und Astronomie, Julius-Maximilians-Universität Würzburg, Würzburg, Germany 175Department of Physics, University of Warwick, Coventry, United Kingdom 176Waseda University, Tokyo, Japan 177D t t f P ti l Ph i W i I tit t f S i R h t I l of Physics and Astronomy, Tufts University, Medford, Massachusetts, USA 167Department of Physics and Astronomy, Tufts University, Medford, Massachusetts, USA 173Department of Physics and Astronomy, University of Victoria, Victoria, British Columbia, Canada 174Fakultät für Physik und Astronomie, Julius-Maximilians-Universität Würzburg, Würzburg, Germany 175Department of Physics, University of Warwick, Coventry, United Kingdom 176Waseda University, Tokyo, Japan 177Department of Particle Physics Weizmann Institute of Science Rehovot Israel 024908-33 PHYSICAL REVIEW C 98, 024908 (2018) 178Department of Physics, University of Wisconsin, Madison, Wisconsin, USA 179Fakultät für Mathematik und Naturwissenschaften, Fachgruppe Physik, Bergische Universität Wuppertal, Wuppertal, Germany 180Department of Physics, Yale University, New Haven, Connecticut, USA 181Yerevan Physics Institute, Yerevan, Armenia PHYSICAL REVIEW C 98, 024908 (2018) M. AABOUD et al. 178Department of Physics, University of Wisconsin, Madison, Wisconsin, USA 179Fakultät für Mathematik und Naturwissenschaften, Fachgruppe Physik, Bergische Universität Wuppertal, Wuppertal, Germany 180Department of Physics, Yale University, New Haven, Connecticut, USA 181Yerevan Physics Institute, Yerevan, Armenia aDeceased. lso at Department of Physics, King’s College London, London, United Kingdom. bAlso at Department of Physics, King’s College London, London, United Kingdom. cAlso at Istanbul University, Dept. of Physics, Istanbul, Turkey. cAlso at Istanbul University, Dept. of Physics, Istanbul, Turkey. lso at Institute of Physics, Azerbaijan Academy of Sciences, Baku, Azerbaijan. lso at TRIUMF Vancouver BC Canada dAlso at Institute of Physics, Azerbaijan Academy of Sciences, Baku, Azerbaijan. eAlso at TRIUMF, Vancouver, BC, Canada. fAlso at Department of Physics and Astronomy, University of Louisville, Louisville, KY, USA. lso at Department of Physics and Astronomy, University of Louisville, Louisville, KY, USA. gAlso at Department of Physics, California State University, Fresno, CA, USA. gAlso at Department of Physics, California State University, Fresno, CA, USA. hAlso at Department of Physics, University of Fribourg, Fribourg, Switzerland. hAlso at Department of Physics, University of Fribourg, Fribourg, Switzerland. iAlso at II. Physikalisches Institut, Georg-August-Universität Göttingen, Göttingen, Germany. iAlso at II. Physikalisches Institut, Georg-August-Universität Göttingen, Göttingen, Germany. jAlso at Departament de Fisica de la Universitat Autonoma de Barcelona, Barcelona, Spain. jAlso at Departament de Fisica de la Universitat Autonoma de Barcelona, Barcelona, Spain. kAlso at Tomsk State University, Tomsk, and Moscow Institute of Physics and Technology State University, Dolgoprudny, Russia. lAlso at The Collaborative Innovation Center of Quantum Matter (CICQM), Beijing, China. y y gy y g p y lAlso at The Collaborative Innovation Center of Quantum Matter (CICQM), Beijing, China. lAlso at The Collaborative Innovation Center of Quantum Matter (CICQM), Beijing, China. lAlso at The Collaborative Innovation Center of Quantum Matter (CICQM), Beijing, China mAlso at Universita di Napoli Parthenope, Napoli, Italy. mAlso at Universita di Napoli Parthenope, Napoli, Italy. nAlso at Institute of Particle Physics (IPP), Victoria, BC, Canada. nAlso at Institute of Particle Physics (IPP), Victoria, BC, Canada. oAlso at Dipartimento di Fisica E. Fermi, Università di Pisa, Pisa, Italy. oAlso at Dipartimento di Fisica E. Fermi, Università di Pisa, Pisa, Italy. pAlso at Horia Hulubei National Institute of Physics and Nuclear Engineering, Bucharest, Romania. pAlso at Horia Hulubei National Institute of Physics and Nuclear Engineering, B qAlso at CPPM, Aix-Marseille Université, CNRS/IN2P3, Marseille, France. qAlso at CPPM, Aix-Marseille Université, CNRS/IN2P3, Marseille, France. PHYSICAL REVIEW C 98, 024908 (2018) at CPPM, Aix-Marseille Université, CNRS/IN2P3, Ma rAlso at Department of Physics, St. Petersburg State Polytechnical University, St. Petersburg, R rAlso at Department of Physics, St. Petersburg State Polytechnical University, St. Petersburg, Russia sAlso at Borough of Manhattan Community College, City University of New York, NY, USA. sAlso at Borough of Manhattan Community College, City University of New York, NY, USA. tAlso at Department of Financial and Management Engineering, University of the Aegean, Chi at Department of Financial and Management Engineeri tAlso at Department of Financial and Management Engineering, University of the Aegean, Chios, Greece. uAlso at Centre for High Performance Computing, CSIR Campus, Rosebank, Cape Town, S uAlso at Centre for High Performance Computing, CSIR Campus, Rosebank, Cape Town, South Africa. vAlso at Louisiana Tech University, Ruston, LA, USA. vAlso at Louisiana Tech University, Ruston, LA, USA. vAlso at Louisiana Tech University, Ruston, LA, USA. y wAlso at Institucio Catalana de Recerca i Estudis Avancats, ICREA, Barcelona, Spain. a de Recerca i Estudis Avancats, ICREA, Barcelona, Spa Also at Institucio Catalana de Recerca i Estudis Avancat xAlso at Department of Physics, University of Michigan, Ann Arbor, MI, USA. xAlso at Department of Physics, University of Michigan, Ann Arbor, MI, USA. yAlso at LAL, Université Paris-Sud, CNRS/IN2P3, Université Paris-Saclay, Orsay, France. yAlso at LAL, Université Paris-Sud, CNRS/IN2P3, Université Paris-Saclay, Orsa zAlso at Graduate School of Science, Osaka University, Osaka, Japan. zAlso at Graduate School of Science, Osaka University, Osaka, Japan. aaAlso at Physikalisches Institut, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany. aaAlso at Physikalisches Institut, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany. abAlso at Institute for Mathematics, Astrophysics and Particle Physics, Radboud University Nijmegen/Nikhef, Nijmegen, Netherlands. acAlso at Near East University, Nicosia, North Cyprus, Mersin, Turkey. abAlso at Institute for Mathematics, Astrophysics and Particle Physics, Radboud University Nijmegen/Nikhef, Nijmegen, Netherlands. acAlso at Near East University, Nicosia, North Cyprus, Mersin, Turkey. , p y y , y j g , j g , acAlso at Near East University, Nicosia, North Cyprus, Mersin, Turkey. acAlso at Near East University, Nicosia, North Cyprus, Mersin, Turkey. adAlso at Institute of Theoretical Physics, Ilia State University, Tbilisi, Georgia. adAlso at Institute of Theoretical Physics, Ilia State University, Tbilisi, Georg aeAlso at CERN, Geneva, Switzerland. afAlso at Manhattan College, New York, NY, USA. agAlso at Hellenic Open University, Patras, Greece. PHYSICAL REVIEW C 98, 024908 (2018) ahAlso at The City College of New York, New York, NY, U ahAlso at The City College of New York, New York, NY, USA. y g aiAlso at Departamento de Física Teorica y del Cosmos, Universidad de Granada, Granada, Spain. o at Departamento de Física Teorica y del Cosmos, Univ ajAlso at Department of Physics, California State University, Sacramento, CA, USA. ajAlso at Department of Physics, California State University, Sacramento, CA, USA. akAlso at Moscow Institute of Physics and Technology State University, Dolgoprudny, Russia. akAlso at Moscow Institute of Physics and Technology State University, Dolgoprudny, alAlso at Département de Physique Nucléaire et Corpusculaire, Université de Genève, Genève, Switzerland. alAlso at Département de Physique Nucléaire et Corpusculaire, Université de Genève, Genève, Département de Physique Nucléaire et Corpusculaire, U amAlso at Department of Physics and Astronomy, University of Shefﬁeld, Shefﬁeld, United Kingdom. amAlso at Department of Physics and Astronomy, University of Shefﬁeld, Shefﬁeld, United Kingdom. hool of Physics, Sun Yat-sen University, Guangzhou, Ch aoAlso at Department of Applied Physics and Astronomy, University of Sharjah, Sharjah, United Arab Emirates aoAlso at Department of Applied Physics and Astronomy, University of S aoAlso at Department of Applied Physics and Astronomy, University of Sharjah apAlso at Institut für Experimentalphysik, Universität Hamburg, Hamburg, Germany. apAlso at Institut für Experimentalphysik, Universität Hamburg, Hamburg, Germany. aqAlso at National Research Nuclear University MEPhI, Moscow, Russia. aqAlso at National Research Nuclear University MEPhI, Moscow, Russia. y arAlso at Institute for Particle and Nuclear Physics, Wigner Research Centre for Physics, Budapest, Hungary. arAlso at Institute for Particle and Nuclear Physics, Wigner Research Centre for Physics, Bud asAlso at Giresun University, Faculty of Engineering, Giresun, Turkey. asAlso at Giresun University, Faculty of Engineering, Giresun, Turkey. atAlso at Department of Physics, Nanjing University, Nanjing, China. atAlso at Department of Physics, Nanjing University, Nanjing, Chin auAlso at Institute of Physics, Academia Sinica, Taipei, Taiwan. o at Institute of Physics, Academia Sinica, Taipei, Taiwa 024908-34	38,359
hal-03225621-Synth%C3%A8se%20des%20r%C3%A9sultats%20de%20l%27%C3%A9tude%20de%20la%20consommation%20en%20Guadeloupe%20%28projet%20NuTWInd%29.txt_1	French-Science-Pile	Open Science	Various open science	2,021	Synthèse des résultats de l'étude de la consommation en Guadeloupe (projet Nutwind). [Rapport de recherche] INRAE. 2021. ⟨hal-03225621⟩	None	French	Spoken	1,840	2,960	Synthèse des résultats de l’étude de la consommation en Guadeloupe (projet Nutwind) Mathilde Gorza, Pascale Bazoche, Valérie Angeon To cite this version: Mathilde Gorza, Pascale Bazoche, Valérie Angeon. Synthèse des résultats de l’étude de la consommation en Guadeloupe (projet Nutwind). [Rapport de recherche] INRAE. 2021. �hal-03225621� HAL Id: hal-03225621 https://hal.science/hal-03225621 Submitted on 16 Jun 2021 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 des résultats de l'étude de la consommation en Guadeloupe (projet NuTWInd) Merci pour votre participation Le projet NuTWInd est un projet de recherche coordonné par l’Institut national de recherche pour l’agriculture, l’alimentation et l’environnement (INRAE) et financé par l’agence nationale de la recherche (ANR). Son objectif principal est de comprendre les relations entre l'offre alimentaire et les préférences des populations pour proposer des stratégies en vue d’améliorer la sécurité nutritionnelle aux Antilles françaises. Dans le cadre de ce projet, une équipe de recherche INRAE a animé des ateliers de consommation au lycée hôtelier du Gosier durant le mois de mars dernier. Au total, 179 volontaires ont participé à ces sessions. Nous vous remercions de votre implication ! L’analyse détaillée des résultats est en cours mais nous tenions à vous proposer une première synthèse et des éléments de réponse au questionnaire auquel vous avez répondu. 1. Petit aperçu des résultats des enquêtes de consommation L’expérience consistait à évaluer différents nectars de goyave et répondre à un questionnaire sur les habitudes alimentaires, les connaissances et les croyances alimentaires, et la situation sociodémographique des Guadeloupéens pour comprendre leurs préférences alimentaires. Vous étiez 40% à avoir moins de 30 ans, 17% entre 30 et 39 ans, 16% entre 40 et 49 ans, 19% entre 50 et 59 ans et 8% de plus de 60 ans. Les jeunes ont représenté volontairement la part la plus importante du panel car plusieurs études ont montré qu’ils sont les plus grands consommateurs de produits sucrés. Comment les consommateurs Guadeloupéens comprennent-ils les informations nutritionnelles ? Depuis 2016, la réglementation européenne a rendu obligatoire les informations nutritionnelles sur la plupart des produits alimentaires. Les consommateurs sont donc régulièrement confrontés à ces informations sur les étiquettes des produits. Nous avons souhaité comprendre comment les consommateurs perçoivent ces informations. Ainsi, plusieurs questions portaient sur la connaissance de termes de nutrition et la lecture d’étiquettes nutritionnelles. Notamment, on peut noter que : - 3% des participants ont répondu que les jus de fruits ne contiennent pas de sucre ajouté. En effet, il faut savoir que les jus de fruits ne peuvent pas contenir de sucre ajouté (voir la partie 2 du document). - 13% ont répondu que l’allégation « allégé en sucre » signifie que le produit contient au minimum 30% de sucre en moins qu’un produit standard, ce qui est le cas. 1 Concernant la lecture de la table nutritionnelle présentée ci-dessous : - 78% des participants ont repéré la quantité correcte de sucres contenue dans 100g de produit. - 47% ont repéré la quantité correcte de lipides (c’est-à-dire de matières grasses) contenue dans 100g de produit. - 72% ont indiqué la valeur énergétique correcte d’une gaufrette. Figure 1 : Étiquette nutritionnelle d’un paquet de gaufrettes goût vanille Les recommandations du Plan National Nutrition Santé (PNNS) sont-elles connues par les consommateurs ? Lancé en 2001, le PNNS est un plan de santé publique visant à améliorer l’état de santé de la population en agissant sur la nutrition qui est un facteur majeur de protection de la santé. Ces recommandations sont actualisées tous les 4 ans selon les objectifs nutritionnels de santé publique. Ainsi, différentes questions portaient sur la connaissance des recommandations incluses dans le PNNS actuel. Le graphique ci-dessous présente pour chaque question posée la réponse correcte correspondante avec la répartition des réponses des participants. Concernant la quantité de consommation maximum de boisson à base de fruits recommandée par jour, 13% des participants ont répondu 10cL par jour, ce qui correspond à la quantité indiquée dans le PNNS. De plus, 70% des participants ont répondu correctement que manger un fruit et boire un verre de jus ne fournit pas les mêmes apports nutritionnels. 2 Taux de réponses aux questions relatives aux recommandations du PNNS Quantité maximum de jus recommandée par jour ? Réponse correcte : 10cL Manger un fruit entier équivaut à boire un verre de jus : FAUX La consommation de jus augmente les risques de diabètes, obésités et maladies cardiovasculaires : VRAI L'eau est la seule boisson recommandée : VRAI Les jus de fruits sont riches en sucres et pauvres en fibres : VRAI Il est recommandé de boire au moins un verre de jus de fruits par jour : FAUX 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% Correct Incorrect Ne sait pas Figure 2 : Taux de réponses correctes/incorrectes aux questions autour des recommandations du PNNS Quelles sont les habitudes de consommation des boissons à base de fruits des Guadeloupéens ? Toute une partie du questionnaire était dédiée aux habitudes alimentaires, plus particulièrement centrées sur les boissons à base de fruits. Le graphique ci-dessous présente la fréquence de consommation de boissons à base de fruits des participants. On peut noter que les 2/3 des participants en consomment au minimum plusieurs fois par semaine. Répartition des fréquences de consommation de boisson à base de fruits Moins d'une fois par mois ou jamais Environ 1 fois par mois Environ 2 à 3 fois par mois Environ 1 fois par semaine Plusieurs fois par semaine Une fois par jour Plusieurs fois par jour 0 5 10 15 20 25 30 35 Pourcentage (%) Figure 3 : Répartition des fréquences de consommation de boisson à base de fruits des participants 3 Une question portait sur la quantité de boisson ingérée par prise, ce qui signifie la quantité bue à chaque fois que le participant consomme ce type de boisson. Quantité de boisson à base de fruit consommée par prise 33% Pour rappel, la quantité maximum recommandée par jour selon le PNNS est de 10cL par jour et pas tous les jours. 39% 7% 2% 19% 10cL 20cL 33cL 50cL 1L Figure 4 : Répartition de la quantité consommée de boisson à base de fruits par prise par les participants Quelles sont les croyances alimentaires qui pourraient influencer les pratiques et les préférences alimentaires des Guadeloupéens ? Pour identifier ce qui peut influencer les choix des consommateurs, nous nous sommes intéressées aux croyances alimentaires de la population guadeloupéenne. Les croyances alimentaires correspondent à ce que l’individu attend du produit et à l’idée qu’il s’en fait : par rapport à son origine, sa qualité, son goût, ses apports pour la santé, etc. Les croyances sont donc subjectives, hétérogènes et propres à un individu. On note que : - 79% des participants pensent que le sport permet de compenser la consommation de produits énergétiques. - 86% voient dans les étiquettes nutritionnelles des informations utiles et pertinentes. - 93% trouvent que les jus faits maison sont de meilleure qualité nutritionnelle que ceux du commerce. - 89% estiment que les sodas « light » ne sont pas bons pour la santé. 4 2. Quelques définitions nécessaires à la lecture d’une étiquette nutritionnelle et présentation des recommandations du PNNS • Des termes de nutrition à connaître pour comprendre les étiquettes nutritionnelles La valeur énergétique : quantité d’énergie pouvant être extraite d’un aliment et fournie à l’organisme. Elle s’exprime en kilojoules (kJ) ou kilocalories (kcal). Les glucides : type de nutriment qui contribue à l’apport énergétique, ils sont constitués de sucres simples et complexes et de fibres. Les matières grasses : type de nutriment aussi appelé « lipides », qui contribue à l’apport énergétique et permet notamment le stockage de l’énergie dans les tissus adipeux. • Qu’est-ce qu’une allégation ? À quoi cela correspond ? Les allégations nutritionnelles et de santé sont des mentions, images ou symboles valorisant les denrées alimentaires sur le plan nutritionnel ou de la santé comme par exemple « jus de fruit riche en vitamine C », « riche en calcium », « source de fibres », « allégé en sucres » ou « bon pour le cœur ». L’allégation « allégé en sucre » signifie que le produit contient au minimum 30% de sucre en moins qu’un produit standard. • Nectar, jus, boissons fruitées : quelles différences ? Les nectars de fruits sont constitués de jus ou de purée de fruits (entre 25% et 50% minimum selon les fruits), d’eau avec ou sans addition de sucre et/ou édulcorant et/ou miel (20% de sucre maximum). Les jus de fruits sont fabriqués à partir de l’extraction du fruit. L’ajout de sucre est interdit depuis 2013. Ce qui différencie un jus et un nectar est donc la teneur en fruit, elle est plus faible dans les nectars de fruits. Différents procédés de fabrication de jus de fruits peuvent être utilisés : - Les purs jus sont obtenus par simple pressage du fruit, sans aucun autre ajout. Les purs jus peuvent être pasteurisés ou stérilisés. - Les jus à base de concentré sont des jus auxquels l'eau est extraite par évaporation sur le lieu même de l’exploitation et dont le concentré ainsi obtenu est congelé. Cette concentration a pour but de diminuer les coûts de stockage et de transport. À son arrivée sur le lieu de conditionnement, le jus est reconstitué en ajoutant la même quantité d’eau qui avait été préalablement enlevée. 5 Une boisson à base de fruits est un mélange d’eau, de sucre et d’un minimum de 10 % de fruits. Ce sont des boissons qui se rapprochent des sodas : elles contiennent souvent des sirops ou des arômes de synthèse et des additifs chimiques. Tableau 1 : Présentation des différents types de boissons aux fruits et leur procédé de fabrication simplifié (source : Gorza, 2021) • Quelles sont les recommandations du Plan National Nutrition Santé (PNNS) ? D’après le PNNS, les jus de fruits sont classés dans les boissons sucrées, au même titre que les sodas même light ou les boissons dites « énergisantes ». Leur consommation doit être limitée le plus possible. Dans tous les cas, il ne faut pas consommer plus d’un verre par jour de 10cL maximum. De manière générale, la seule boisson recommandée est l’eau qui peut aussi être apportée par le thé non sucré, le café (sans excès) non sucré et des infusions non sucrées. La consommation de fruits entiers est préférable à celle de jus de fruits pour favoriser l’apport en fibres qui sont très peu présentes dans les jus. Pour plus d’informations, nous vous invitons à consulter l’ensemble des recommandations du PNNS : https://www.mangerbouger.fr/Les-recommandations L’équipe comportement du consommateur, Mathilde Gorza, Pascale Bazoche, Valérie Angeon, INRAE Contacts : pascale.bazoche@inrae.fr / valerie.angeon@inrae.fr 6.	22,017
https://openalex.org/W2999300644	OpenAlex	Open Science	CC-By	2,020	The genetics of intellectual disability: advancing technology and gene editing	Muhammad Ilyas	English	Spoken	7,673	13,331	The genetics of intellectual disability: advancing technology and gene editing [version 1; peer review: 2 approved] Muhammad Ilyas 1,2, Asif Mir1, Stephanie Efthymiou 2, Henry Houlden 2 1Department of Neuromuscular Disorders, UCL Institute of Neurology Queen Square, UCL collage london, Queen Square, London, WC1N 3BG, UK 2Department of Biological Sciences, International islamic university Islamabad, Islamabad, 44000, Pakistan * Equal contributors Open Peer Review Approval Status 1 2 version 1 16 Jan 2020 First published: 16 Jan 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 Latest published: 16 Jan 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 v1 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 Abstract Intellectual disability (ID) is a neurodevelopmental condition affecting Intellectual disability (ID) is a neurodevelopmental condition affecting 1–3% of the world’s population. Genetic factors play a key role causing the congenital limitations in intellectual functioning and adaptive behavior. The heterogeneity of ID makes it more challenging for genetic and clinical diagnosis, but the advent of large-scale genome sequencing projects in a trio approach has proven very effective. However, many variants are still difficult to interpret. A combined approach of next-generation sequencing and functional, electrophysiological, and bioinformatics analysis has identified new ways to understand the causes of ID and help to interpret novel ID- causing genes. This approach offers new targets for ID therapy and increases the efficiency of ID diagnosis. The most recent functional advancements and new gene editing techniques involving the use of CRISPR–Cas9 allow for targeted editing of DNA in in vitro and more effective mammalian and human tissue-derived disease models. The expansion of genomic analysis of ID patients in diverse and ancient populations can reveal rare novel disease-causing genes. Hao Hu, Guangzhou Women and Children’s Medical Center, Guangzhou, China Max-Planck Institute for Molecular Genetics, Berlin, Germany 1. Jennifer Winter, Institute of Human Genetics, University Medical Center of the Johannes Gutenberg University, Mainz, Germany 2. Any comments on the article can be found at the end of the article. y Intellectual Disability, Neurological Disorders, Gene Editing, Mental Retrdation, NGS, WES, CRISPR/Cas9. Keywords y Intellectual Disability, Neurological Disorders, Gene Editing, Mental Retrdation, NGS, WES, CRISPR/Cas9. Intellectual Disability, Neurological Disorders, Gene Editing, Mental Retrdation, NGS, WES, CRISPR/Cas9. Jennifer Winter, Institute of Human Genetics, University Medical Center of the Johannes Gutenberg University, Mainz, Germany 2. Page 1 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 Corresponding authors: Asif Mir (asif.mir@iiu.edu.pk), Henry Houlden (h.houlden@ucl.ac.uk) Author roles: Ilyas M: Writing – Original Draft Preparation; Mir A: Methodology, Writing – Review & Editing; Efthymiou S: Writing – Review & Editing; Houlden H: Funding Acquisition, Supervision, Writing – Review & Editing Competing interests: No competing interests were disclosed. Grant information: The authors’ work is funded by the MRC (MR/S01165X/1, MR/S005021/1, G0601943), The National Institute for Health Research University College London Hospitals Biomedical Research Centre, Rosetree Trust, Ataxia UK, MSA Trust, Brain Research UK, Sparks GOSH Charity, Muscular Dystrophy UK (MDUK), Muscular Dystrophy Association (MDA USA), and Higher Education Commission of Pakistan (Project 7028_NRPU). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2020 Ilyas M 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 work is properly cited. How to cite this article: Ilyas M, Mir A, Efthymiou S and Houlden H. The genetics of intellectual disability: advancing technology and gene editing [version 1; peer review: 2 approved] F1000Research 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 First published: 16 Jan 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 Corresponding authors: Asif Mir (asif.mir@iiu.edu.pk), Henry Houlden (h.houlden@ucl.ac.uk) Author roles: Ilyas M: Writing – Original Draft Preparation; Mir A: Methodology, Writing – Review & Editing; Efthymiou S: Writing – Review & Editing; Houlden H: Funding Acquisition, Supervision, Writing – Review & Editing Competing interests: No competing interests were disclosed. Grant information: The authors’ work is funded by the MRC (MR/S01165X/1, MR/S005021/1, G0601943), The National Institute for Health Research University College London Hospitals Biomedical Research Centre, Rosetree Trust, Ataxia UK, MSA Trust, Brain Research UK, Sparks GOSH Charity, Muscular Dystrophy UK (MDUK), Muscular Dystrophy Association (MDA USA), and Higher Education Commission of Pakistan (Project 7028_NRPU). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2020 Ilyas M et al. Introduction is liable to mutation, mediating genetic plasticity. The expan- sion of tandem repeats can cause a range of disorders associ- ated with ID such as X-linked ID (XLID)9, various ataxias, motor neuron disease, and epilepsy. Emerging data suggest that tandem repeat polymorphisms (TRPs) can contribute to the missing heritability of polygenic disorders10–13. Furthermore, various metabolic factors, repeat expansions of nucleotides, and mitochondrial DNA variants can also contribute to ID7. Environ- mental factors such as hazardous chemical exposures, infections during pregnancy, and UV radiation are also reported to cause ID. In addition to these, lack of nutrition, cultural deprivation, childhood diseases such as measles, meningitis, and severe head injury can cause malfunction of the nervous system, lead- ing to ID14. Even though the etiological factors of ID are very broad, as shown in Figure 2, in 50% of individuals the cause of ID is still unknown, but the most prominent causes are genetic15. Intellectual disability (ID) occurs in the developmental period before the age of 18 years. ID is a heterogeneous group of disorders characterized by significantly impaired intellectual functioning and deficits in adaptive behaviors1. It affects 1–3% of the world population, is the most common developmental disorder, and represents an important socio-economic problem in healthcare. However, owing to the heterogeneity of ID, its frequency ratio changes worldwide2. Diagnosis of ID is also performed with the identification of clinical phenotype symptoms such as delayed speech, hypotonia, and seizures3. In the past decade, the genetic background of ID was believed to be mostly autosomal dominant (de novo mutations) in the outbred countries such as the USA and those in Western Europe, while in the middle-east countries where inbreeding is common, autosomal recessive ID has some preponderance4,5. Next-generation sequencing (NGS) provided tremendous power to sequence personal genomes and detect a large number of genetic variants. The discovery of disease-causing variants by whole exome or genome sequencing in patients has dramati- cally changed our perspective on precision medicine6. Through NGS methods, now it is possible to find pathogenic mutations, including novel mutations, associated with ID7. A combined NGS and bioinformatics approach is used to iden- tify novel ID genes and screen candidate ID genes as well. NGS has efficiently expedited ID research and provided new strategies at the clinical level in the last few years, as shown in Figure 1. Etiological classification of intellectual disability Etiological classification of intellectual disability Multiple factors are involved in causing ID. Genetic factors include genetic variations such as aneuploidies, copy number variations (CNVs), and tandem repeats in specific genes8. DNA Keywords 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 work is properly cited. How to cite this article: Ilyas M, Mir A, Efthymiou S and Houlden H. The genetics of intellectual disability: advancing technology and gene editing [version 1; peer review: 2 approved] F1000Research 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 First published: 16 Jan 2020, 9(F1000 Faculty Rev):22 https://doi.org/10.12688/f1000research.16315.1 Corresponding authors: Asif Mir (asif.mir@iiu.edu.pk), Henry Houlden (h.houlden@ucl.ac.uk) Page 2 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 Introduction Genomics England’s PanelApp database (https://panelapp.genomicsengland.co.uk/panels/285/) shows that around 1,396 genes cause ID (see Table 1, Extended data). Cytogenetic abnormalitiesi The identification of genetic factors causing ID has advanced in terms of number and type with recent developments in cytoge- netic techniques. The first genetic test used in the investigation of ID was karyotyping16 to identify aneuploidies, such as Down’s syndrome (trisomy 21) and Edwards syndrome (trisomy 18), and large structural re-arrangements such as insertions, deletions, and duplications17,18. Karyotyping detects only large dele- tions or insertions owing to its low resolution. Fluorescence in situ hybridization (FISH) is used to detect structural abnor- malities and numerical changes on chromosomes. Specific probes are used in FISH for the analysis of chromosomal aber- ration19. A number of studies on ID describe the growing benefits of using NGS in clinics for diagnostic purposes20. Chromosomal aberrations The mortality rate is very high with this condition; only 5–10% of affected children survive after the first year of life24. Patau syndrome is a trisomy of chromosome 13, characterized by malformations of the central nervous sys- tem (CNS). The occurrence of this syndrome is 1 in 12,000 in the general population23. The survival rate is very low in infants with trisomy 13, but it depends on the severity of the condi- tion in infants, i.e. whether or not there are cerebral, cardiac, or other congenital malformations25. the most frequent form of ID. The clinical features of Down’s syndrome include dysmorphic features, seizures, psychomo- tor slowing, and congenital malformation. These conditions may not be present in each affected individual22. Edwards syndrome is a trisomy of chromosome 18 characterized by psychomo- tor and cognitive brain impairment, malformation, and growth deficiency in infants23. The occurrence of Edwards syndrome is 1 out of 8,000 live births. The mortality rate is very high with this condition; only 5–10% of affected children survive after the first year of life24. Patau syndrome is a trisomy of chromosome 13, characterized by malformations of the central nervous sys- tem (CNS). The occurrence of this syndrome is 1 in 12,000 in the general population23. The survival rate is very low in infants with trisomy 13, but it depends on the severity of the condi- tion in infants, i.e. whether or not there are cerebral, cardiac, or other congenital malformations25. Chromosomal aberrations Chromosomal aberrations An extra copy of chromosome 21 as a result of an error in cell division can cause a trisomy known as Down’s syndrome21. It is Figure 1. The number of articles on intellectual disability published in the last seven years identified using the PubMed search terms “ID”, “mental retardation”, “next-generation sequencing”, and “exome sequencing”. Figure 1. The number of articles on intellectual disability published in the last seven years identified using the PubMed search terms “ID”, “mental retardation”, “next-generation sequencing”, and “exome sequencing”. Page 3 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 Figure 2. Intellectual disability classification. Multiple factors are involved in intellectual disability including genetic inheritance and environmental conditions. ility classification. Multiple factors are involved in intellectual disability including genetic inheritance and Figure 2. Intellectual disability classification. Multiple factors are involved in intellectual disability including genetic inheritance and environmental conditions. Figure 2. Intellectual disability classification. Multiple factors are involved in intellectual disability inclu environmental conditions. on a large cohort reported 118 rare de novo CNVs30. Analysis of these reported CNVs and candidate genes pinpointed 10 genes with loss of function associated with ID31. CNVs cannot be vis- ibly detected with a light microscope. Array comparative genomic hybridization (CGH) can perform rapid genome-wide analysis at a high resolution and detect CNVs, gain or loss, at the chromosomal level32. Array CGH analysis of unresolved ID cases increased the identification of pathogenic CNVs up to 13% in recent years. Phenotypes associated with these 13% of cases are congenital defects, primary microcephaly, and short stature33. A disorder (Online Mendelian Inheritance in Man [OMIM] #612001) with microdeletion on 15q13.3 shows an ID phenotype with a complex variety of seizures, autism, and psychiatric conditions34,35. SNP array is also a new technique as compared to array CGH which can detect CNVs36. A novel genetic disorder related to ID with a 3q29 microdeletion was reported using microarray techniques37. the most frequent form of ID. The clinical features of Down’s syndrome include dysmorphic features, seizures, psychomo- tor slowing, and congenital malformation. These conditions may not be present in each affected individual22. Edwards syndrome is a trisomy of chromosome 18 characterized by psychomo- tor and cognitive brain impairment, malformation, and growth deficiency in infants23. The occurrence of Edwards syndrome is 1 out of 8,000 live births. Whole exome and genome sequencing h l i i CNVs are small segments of DNA that vary in number. Usu- ally each individual carries two copies: one that comes from the maternal side and one from the paternal side. Variations in copy number occur through duplications and deletions of the small DNA segments. But not all copy numbers, that are either a dele- tion or duplication, are pathogenic to humans26,27. The largest database for CNVs (DGV) is available online (www.dgv.tcag. ca)28. Decipher is another database used by clinicians to com- pare clinical and genetic information for the identification and interpretation of pathogenic variants (sequence variants and copy number variants) in patients with ID29. CNVs causing de novo and inherited mutations have been associated with ID. A study Whole exome sequencing (WES) is an efficient technology that can increase the diagnostic yield when searching for alleles causing rare Mendelian disorders. Exome analysis examines the protein’s encoding region, where an estimated 85% of disease- causing mutations are believed to occur38. It has been an invaluable tool in gene discovery for ID. WES performed on three members of a family with autosomal dominant ID (MRD44; 617061) showed a heterozygous mutation, a 1 bp deletion (c.4466delA, NM_007118) in exon 30 of the TRIO gene. This mutation resulted in a framshift and premature termination (Gln1489ArgfsTer11) in the GEFD1 domain. A Page 4 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 9-year-old girl with autosomal dominant ID was also found to carry a de novo heterozygous c.3239A-T transversion (c.3239A-T, NM_007118) in exon 19 of the TRIO gene, which causes changes in the spectrin repeat domain39. Whole genome sequencing (WGS) allows examination of single- nucleotide variants (SNVs), indels, structural variants (SVs), and CNVs in both the ~1% part of the genome that encodes protein sequences and the ~99% of remaining non-coding sequences. Therefore, WGS has more reliable sequence coverage with more uniformity. It is likely to reveal many novel variants and genes and derive new scientific and clinical findings for ID. With the rapid drop in sequencing cost and the ability of WGS to rapidly produce large volumes of data, it is becoming a powerful tool for genomic research. genes have been linked with ID50. X-linked recessive fragile X syndrome occurs on chromosome Xq27.3 in the FMR1 gene at the 5' untranslated region (UTR) owing to the expansion of CGG trinucleotide repeats51. Autosomal recessive intellectual disability Autosomal recessive ID is a genetically heterogeneous group of disorders41. Autosomal recessive ID occurs in syndromic and non-syndromic forms. The syndromic type of ID is character- ized by intellectual problems occurring with a group of other phenotypic features42. Non-syndromic ID is characterized by a lack of associated pathology. Genes linked with non-syndromic ID are being studied to understand the normal variation in intelligence43. Distinction in intelligence quotient (IQ) is linked with those genes that can also cause large variations in intellectual ability when mutated. Homozygosity mapping is performed to check the autosomal recessive causes of ID in con- sanguineous families with affected siblings44. OMIM and SysID (http://sysid.cmbi.umcn.nl/) search results show that 399 genes can cause autosomal recessive ID (see Table 2, Extended data). In vitro and in vivo study of intellectual disability Biological assays can be used to study any undefined variant and its role or pathogenicity. These assays can be used in mutated cells and also directly in patient-derived cells58. To identify the pathogenicity of candidate ID genes, electrophysiological studies of SH-SY5Y neuronal cell lines are a very powerful approach59 to capture early cortical development with high fidelity, thus helping to study genes that are relevant in early cortical develop- ment and associated with ID60. In vivo studies of candidate ID genes provide additional information regarding pathogenicity. Different animal models can be used to study ID genes. CRBN knockout (CrbnKO) mice have been used to study learning and memory tasks. Loss of CRBN results in memory problems, learning problems when AMPK activity is accelerated, block- ing of mTORC1 signaling, and a decreased level of glutama- tergic synaptic proteins. These findings show that the CrbnKO mouse is an ideal animal model to understand the molecular mechanisms of learning and memory problems in ID patients61. Zebrafish can also be used as a model organism to study the function of genes associated with ID. Zebrafish have unique features including the rapid development of embryos and easy visualization of the nervous system during developmental stages, which make them an ideal organism to study the func- tion of genes. Overexpression comparison can be performed between the wild-type PK1A gene and the zebrafish ortholog of PK1A. In this case, zebrafish mutants show severe phenotypes as compared to the wild-type, and a mutant PK1A gene has been reported in patients with epilepsy. These results signify that the mutation changes the in vivo function of PK1A62. Autosomal dominant intellectual disability The inheritance pattern of autosomal dominant ID is when an indi- vidual carries one copy of a mutant allele and one normal allele on a gene. Autosomal dominant ID is caused by heterozygous mutations in different reported genes and CNVs. Tuberous scle- rosis, neurofibromatosis, and myotonic dystrophy are autosomal dominant disorders linked with ID45. SNP microarray analy- sis of the methyl binding domain gene on chromosome 2q23.1 showed that a 200 kb deletion in exon 6 of a female patient was associated with autosomal dominant ID46. It is difficult to find the estimated frequency of mutations in autosomal dominant ID genes. ARID1B, SYNGAP1, DYRK1A, MED13L, KCNQ2, CTNNB1, STXB1, KMT2A, PACS1, FOXP1, and SMARCA2 are the most commonly mutated autosomal dominant ID genes47,48. Some of these genes are essential for neuronal differentiation in the developing brain and play important roles in synaptic forma- tion and transmission49. The outcomes of our OMIM search show that in total around 180 genes or loci, as reported in the literature, are involved in autosomal dominant ID (see Table 3, Extended data). Whole exome and genome sequencing h l i i Fragile X syndrome is clinically characterized as ID; phenotypically, patients show dysmorphic facial features and protruded ears. FMR1 encodes for a protein that provides RNA stability and plays a key role in brain devel- opment and neuronal plasticity. Deficiency of the FMRP pro- tein causes suppression or excitation of GABA that results in low synaptic connections, leading to syndromic features in patients52. The number of new X-linked ID genes identified has increased rapidly over the last few years with the use of NGS. More than 140 known X-linked genes have been reported53,54. Autosomal recessive intellectual disability In addition, the TAF1 gene is associated with ID. Functional study of this gene Identification of candidate and novel intellectual disability genes Inherited mutations causing intellectual disability Single gene disorders are grouped into different types on the basis of inheritance pattern40. Different techniques have been used to identify novel ID-causing genes over the past few decades. For the identification of novel or candidate genes in affected families, it is necessary to recon- struct the family pedigree and perform some clinical investigation before reaching any conclusion55,56. The introduction of robust microarray technologies in research has increased the power of identification; SNP arrays can detect small deletions and micro duplications in the probands57. NGS has accelerated the speed of identification of novel ID-causing genes. NGS technology has become very popular over the past 5 years, with a consider- able number of new ID genes and candidate genes reported. NGS can detect SNVs and small insertions/deletions in the whole genome. X-linked intellectual disability The human X-chromosome comprises 5% of the human genome, but an increasing number of genetic diseases are associated with the X chromosome; approximately 10–12% of X-chromosome Page 5 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 was performed by using a zebrafish knockout model. Severe phenotypes were observed during embryogenesis and neurode- velopment in the zebrafish TAF1 knockout model63. The hetero- geneity of ID makes it very difficult to validate candidate genes as causative ID genes, but in vitro and in vivo studies provide grounds for conclusively identifying certain genes. Single-cell transcriptomics and quantitative proteomics can also be used to improve our understanding of the global changes in the central nervous system when the genome-edited organism is available. been used on fragile X syndrome caused by the extension of tri- nucleotide repeats, which results in deteriorated levels of FMR1 protein and Huntington disease (HD) models. No treatment is avail- able for these conditions65. Induced pluripotent stem cells (iPSCs) of patients with fragile X syndrome in the FRX gene upstream of the CGH repeat were targeted with expressed CRISPR–Cas9 nuclease along with single guide RNA (sgRNA) and resulted in the reactivation of the FMR1 protein66. Further studies show that two sgRNA-guided methods that flanked the trinucleotide CGG repeat produced two double-stranded breaks and the recombina- tion of the breaks resulted in the deletion of repeats. The deletion of repeats resulted in an increased FMR1 protein level67. HD is caused by trinucleotide repeat extensions in the coding region of the huntingtin gene (HTT)68. The CRISPR–Cas9 system has been used recently in the treatment of HD. iPSCs derived from a HD patient were corrected by using CRISPR–Cas9 that selec- tively inactivated the mutant HTT gene without altering the nor- mal allele of the same gene69,70. HD patient fibroblast cell lines were used to show that CAG repeats can be precisely excised using CRISPR–Cas9 nickase from the HTT gene71. Strict guidelines must be adopted to avoid the exploitation of the safety and secu- rity weakness in genome editing techniques as well as to reduce the risk of off-site editing of genome and epigenetic changes with the help of further research. Adopting appropriate biosafety levels for genome editing to stop contamination is important CRISPR–Cas9 gene editing tooli The identification of causal genes and their functional analysis requires further understanding of disease mechanisms, especially for potential therapies, even if we do not have a full understand- ing of the biological functions. Acknowledging this, clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 is an RNA control nuclease system that has become essential for gene editing and correcting mutated genes. It provides potential treatment options for genetic disorders that cause ID (Figure 3). This system has been applied recently to mammalian genomes to stop the expression of the disease gene or to edit mutated genes, thereby correcting the mutation. CRISPR–Cas9 technol- ogy will possibly cure diseases that have no treatment option available, such as trinucleotide repeat expansion diseases causing neurological disorders64. CRISPR–Cas9 systems have Figure 3. The CRISPR–Cas9 system used to correct extension repeats in the huntingtin gene (HTT) by using single-guide RNA (sgRNA) on both sides of the repeats and Cas9 nuclease, creating nicks and removing the CAG repeats, blocking further extension. Figure 3. The CRISPR–Cas9 system used to correct extension repeats in the huntingtin gene (HTT) by using single-guide RNA (sgRNA) on both sides of the repeats and Cas9 nuclease, creating nicks and removing the CAG repeats, blocking further extension. Page 6 of 10 Page 6 of 10 F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 but so is applying rules to cover the biosecurity of gene editing72. Further studies are needed before the application of gene therapy to the treatment of these diseases. Huntington’s disease; ID, intellectual disability; iPSC, induced pluripotent stem cell; NGS, next-generation sequencing; OMIM, Online Mendelian Inheritance in Man; sgRNA, single guide RNA; SNV, single nucleotide variation; WES, whole exome sequencing. Harvard Dataverse: The Genetics of Intellectual Disability, https://doi.org/10.7910/DVN/AEKQOL76 This project contains the following extended data: This project contains the following extended data: This project contains the following extended data: • Table 2: List of Genes causing Autosomal Recessive Intellectual Disability Harvard Dataverse: Replication Data for: The Genetics of Intellectual Disability, https://doi.org/10.7910/DVN/9BJDI677 The genomic understanding of ID has primarily come from devel- oped countries, and our knowledge in developing countries is very limited. This is important, as many of these countries have a young population and culturally prefer large families, indi- cating that the burden of ID and other childhood disorders will increase before technology is developed enough to treat these conditions. Therefore, early diagnosis, education, and genetic counseling will be important for families suffering with these con- ditions. In addition, many of the genes in developing countries are likely to have founder effects that may be amenable to diag- nosis in cultural groups and demographic areas, accelerating diagnosis and revealing carrier status to help family planning. This project contains the following extended data: • Table 3: List of Autosomal Dominant Genes in Omim and PubMed DataBase Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). 2. Harripaul R, Vasli N, Mikhailov A, et al.: Mapping autosomal recessive intellectual disability: combined microarray and exome sequencing identifies 26 novel candidate genes in 192 consanguineous families. Mol Psychiatry. 2018; 23(4): 973–984. PubMed Abstract \| Publisher Full Text 1. Bertelli MO, Munir K, Harris J, et al.: “Intellectual developmental disorders”: reflections on the international consensus document for redefining “mental retardation-intellectual disability” in ICD-11. Adv Ment Health Intellect Disabil. 2016; 10(1): 36–58. P bMed Abstract \| P blisher F ll Te t \| Free F ll Te t 3. Redin C, Gérard B, Lauer J, et al.: Efficient strategy for the molecular diagnosis of intellectual disability using targeted high-throughput sequencing. Abbreviations The authors would like to thank the participants and their families for their essential help with this work. They also thank the National Institute for Health Research Biomedical Research Centre at UCLH/UCL. CGH, comparative genomic hybridization; CNV, copy number variation; CRISPR, clustered regularly interspaced short palin- dromic repeats; FISH, fluorescence in situ hybridization; HD, F1000 recommended 2016; 10(1): 36 58. PubMed Abstract \| Publisher Full Text \| Free Full Text Conclusions and the future direction of intellectual disability genetics Data availability Extended data Harvard Dataverse: Replication Data for: The Genetics of Intellectual Disability Review Tables, https://doi.org/10.7910/ DVN/MMUNLR75 The prevalence of ID varies from country to country, and it is especially low in developed countries as compared to less- developed countries73,74. The identification of genes causing ID rapidly increased over the past 3 to 5 years owing to the use of sophisticated sequencing techniques. The diagnosis of ID patients became easy with new massively parallel sequencing methods and the help of different human variant databases. The heteroge- neity of ID makes it difficult for etiological diagnosis; however, WES is likely to be used as the first-line test for ID probands. NGS improves our understanding of the genetic origin of ID. Not all ID-causing genes have been identified yet, but a combined approach of sequencing techniques, functional analysis, and bioin- formatics will help to identify new ID-causing genes. This approach will potentially provide a new way of treating ID. Although ID genes can be therapeutically targeted using the CRISPR–Cas-9 system, this method and its application in mammals is still emerging, and many regulatory, methodological, and off-target effects still need to be understood. This project contains the following extended data: This project contains the following extended data: • Table 1: Estimated total number of genes involved in Intellectual Disability Genes from Literature search and from genome England Database Harvard Dataverse: The Genetics of Intellectual Disability, https://doi.org/10.7910/DVN/AEKQOL76 Harvard Dataverse: The Genetics of Intellectual Disability, https://doi.org/10.7910/DVN/AEKQOL76 References J Med Genet. 2014; 51(11): 724–736. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 4. Kahrizi K, Hu H, Hosseini M, et al.: Effect of inbreeding on intellectual disability revisited by trio sequencing. Clin Genet. 2019; 95(1): 151–159. PubMed Abstract \| Publisher Full Text \| F1000 Recommendation 5. Hu H, Kahrizi K, Musante L, et al.: Genetics of intellectual disability in consanguineous families. Mol Psychiatry. 2019; 24(7): 1027–1039. PubMed Abstract \| Publisher Full Text \| F1000 Recommendation 6. 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MacDonald JR, Ziman R, Yuen RK, et al.: The Database of Genomic Variants: a curated collection of structural variation in the human genome. Nucleic Acids Res. 2014; 42(Database issue): D986–D992. PubMed Abstract \| Publisher Full Text \| Free Full Text 51. Bagni C, Tassone F, Neri G, et al.: Fragile X syndrome: causes, diagnosis, mechanisms, and therapeutics. J Clin Invest. 2012; 122(12): 4314–22. PubMed Abstract \| Publisher Full Text \| Free Full Text 6 9 658 PubMed Abstract \| Publisher Full Text \| Free Full Text 35. Ben-Shachar S, Lanpher B, German JR, et al.: Microdeletion 15q13.3: a locus with incomplete penetrance for autism, mental retardation, and psychiatric disorders. J Med Genet. 2009; 46(6): 382–388. PubMed Abstract \| Publisher Full Text \| Free Full Text 12. Ishiura H, Doi K, Mitsui J, et al.: Expansions of intronic TTTCA and TTTTA repeats in benign adult familial myoclonic epilepsy. Nat Genet. 2018; 50(4): 581–590. PubMed Abstract \| Publisher Full Text \| F1000 Recommendation ; ( ) PubMed Abstract \| Publisher Full Text \| Free Full Text 36. Bernardini L, Alesi V, Loddo S, et al.: High-resolution SNP arrays in mental retardation diagnostics: how much do we gain? Eur J Hum Genet. 2010; 18(2): 178–85. PubMed Abstract \| Publisher Full Text \| Free Full Text 178 85. PubMed Abstract \| Publisher Full Text \| Free Full Text 37. Digilio MC, Bernardini L, Mingarelli R, et al.: 3q29 Microdeletion: a mental retardation disorder unassociated with a recognizable phenotype in two mother-daughter pairs. Am J Med Genet A. 2009; 149(8): 1777–1781. PubMed Abstract \| Publisher Full Text PubMed Abstract \| Publisher Full Text \| F1000 Recommendation 13. Kim K, Bang S, Yoo D, et al.: De novo emergence and potential function of human-specific tandem repeats in brain-related loci. Hum Genet. 2019; 138(6): 661–672. PubMed Abstract \| Publisher Full Text \| F1000 Recommendation PubMed Abstract \| Publisher Full Text \| F1000 Recommendation 14. Winnepenninckx B, Rooms L, Kooy RF: Mental retardation: a review of the genetic causes. Br J Dev Disabil. 2003; 49(96): 29–44. Publisher Full Text 38. Choi M, Scholl UI, Ji W, et al.: Genetic diagnosis by whole exome capture and massively parallel DNA sequencing. Proc Natl Acad Sci U S A. 2009; 106(45): 19096–19101. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 15. Milani D, Ronzoni L, Esposito S: Genetic Advances in Intellectual Disability. J Pediatr Genet. 2015; 4(3): 125–127. PubMed Abstract \| Publisher Full Text \| Free Full Text 16. Tjio JH, Levan A: The chromosome number of man. Hereditas. 1956; 42(1–2): 1–6. Publisher Full Text 39. Pengelly RJ, Greville-Heygate S, Schmidt S, et al.: Mutations specific to the Rac-GEF domain of TRIO cause intellectual disability and microcephaly. J Med Genet. 2016; 53(11): 735–742. PubMed Abstract \| Publisher Full Text \| Free Full Text 17. Raymond FL, Tarpey P: The genetics of mental retardation. Hum Mol Genet. 2006; 15 Spec No 2: R110–R116. PubMed Abstract \| Publisher Full Text ; PubMed Abstract \| Publisher Full Text 56. van Karnebeek CD, Stockler S: Treatable inborn errors of metabolism causing intellectual disability: a systematic literature review. Mol Genet Metab. 2012; 105(3): 368–381. PubMed Abstract \| Publisher Full Text 68. Bates GP, Dorsey R, Gusella JF, et al.: Huntington disease. Nat Rev Dis Primers. 2015; 1: 15005. PubMed Abstract \| Publisher Full Text 64. Cong L, Ran FA, Cox D, et al.: Multiplex genome engineering using CRISPR/Cas systems. Science. 2013; 339(6121): 819–23. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation ( ) PubMed Abstract \| Publisher Full Text \| Free Full Text 29. 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Park CY, Halevy T, Lee DR, et al.: Reversion of FMR1 Methylation and Silencing by Editing the Triplet Repeats in Fragile X iPSC-Derived Neurons. Cell Rep. 2015; 13(2): 234–41. PubMed Abstract \| Publisher Full Text 54. Neri G, Schwartz CE, Lubs HA, et al.: X-linked intellectual disability update 2017. Am J Med Genet A. 2018; 176(6): 1375–1388. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 67. Xie N, Gong H, Suhl JA, et al.: Reactivation of FMR1 by CRISPR/Cas9-Mediated Deletion of the Expanded CGG-Repeat of the Fragile X Chromosome. PLoS One. 2016; 11(10): e0165499. PubMed Abstract \| Publisher Full Text \| Free Full Text 55. Ropers HH: Genetics of early onset cognitive impairment. Annu Rev Genomics Hum Genet. 2010; 11: 161–87. PubMed Abstract \| Publisher Full Text ( ) PubMed Abstract \| Publisher Full Text 69. Shin JW, Kim KH, Chao MJ, et al.: Permanent inactivation of Huntington’s disease mutation by personalized allele-specific CRISPR/Cas9. 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Bavley CC, Rice RC, Fischer DK, et al.: Rescue of Learning and Memory Deficits in the Human Nonsyndromic Intellectual Disability Cereblon Knock- Out Mouse Model by Targeting the AMP-Activated Protein Kinase-mTORC1 Translational Pathway. J Neurosci. 2018; 38(11): 2780–2795. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 73. Maulik PK, Mascarenhas MN, Mathers CD, et al.: Prevalence of intellectual disability: a meta-analysis of population-based studies. Res Dev Disabil. 2011; 32(2): 419–436. PubMed Abstract \| Publisher Full Text \| 74. Westerinen H, Kaski M, Virta LJ, et al.: The nationwide register-based prevalence of intellectual disability during childhood and adolescence. J Intellect Disabil Res. 2017; 61(8): 802–809. Editorial Note on the Review Process Faculty Reviews are review articles written by the prestigious Members of Faculty Opinions. The articles are commissioned and peer reviewed before publication to ensure that the final, published version is comprehensive and accessible. The reviewers who approved the final version are listed with their names and affiliations. Current Peer Review Status: Current Peer Review Status: F1000Research 2020, 9(F1000 Faculty Rev):22 Last updated: 01 AUG 2023 ( ) PubMed Abstract \| Publisher Full Text PubMed Abstract \| Publisher Full Text \| F1000 Recommendation 75. Ilyas M: Replication Data for: The Genetics of Intellectual Disability Review Tables. Harvard Dataverse, V1. 2019. http://www.doi.org/10.7910/DVN/MMUNLR 76. Ilyas M: The Genetics of Intellectual Disability. Harvard Dataverse, V1. 2019. http://www.doi.org/10.7910/DVN/AEKQOL 77. Ilyas M: Replication Data for: The Genetics of Intellectual Disability. Harvard Dataverse, V1. 2019. http://www.doi.org/10.7910/DVN/9BJDI6 62. Bassuk AG, Wallace RH, Buhr A, et al.: A homozygous mutation in human PRICKLE1 causes an autosomal-recessive progressive myoclonus epilepsy- ataxia syndrome. Am J Hum Genet. 2008; 83(5): 572–581. PubMed Abstract \| Publisher Full Text \| Free Full Text 63. Gudmundsson S, Wilbe M, Filipek-Górniok B, et al.: TAF1, associated with intellectual disability in humans, is essential for embryogenesis and regulates neurodevelopmental processes in zebrafish. Sci Rep. 2019; 9(1): 10730. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 64. Cong L, Ran FA, Cox D, et al.: Multiplex genome engineering using CRISPR/Cas systems. Science. 2013; 339(6121): 819–23. PubMed Abstract \| Publisher Full Text \| Free Full Text \| F1000 Recommendation 77. Ilyas M: Replication Data for: The Genetics of Intellectual Disability. Harvard Dataverse, V1. 2019. http://www.doi.org/10.7910/DVN/9BJDI6 77. Ilyas M: Replication Data for: The Genetics of Intellectual Disability. Harvard Dataverse, V1. 2019. http://www.doi.org/10.7910/DVN/9BJDI6 Page 9 of 10 Page 9 of 10 The reviewers who approved this article are: The reviewers who approved this article are: Version 1 Jennifer Winter Institute of Human Genetics, University Medical Center of the Johannes Gutenberg University, Mainz, Germany Competing Interests: No competing interests were disclosed. 1. Hao Hu 1 Guangzhou Women and Children’s Medical Center, Guangzhou, China 2 Max-Planck Institute for Molecular Genetics, Berlin, Germany Competing Interests: No competing interests were disclosed. 2. The benefits of publishing with F1000Research: Your article is published within days, with no editorial bias • You can publish traditional articles, null/negative results, case reports, data notes and more • The peer review process is transparent and collaborative • Your article is indexed in PubMed after passing peer review • Dedicated customer support at every stage • For pre-submission enquiries, contact research@f1000.com Version 1 Version 1 Jennifer Winter Institute of Human Genetics, University Medical Center of the Johannes Gutenberg University, Mainz, Germany Competing Interests: No competing interests were disclosed. 1. Hao Hu 1 Guangzhou Women and Children’s Medical Center, Guangzhou, China 2 Max-Planck Institute for Molecular Genetics, Berlin, Germany Competing Interests: No competing interests were disclosed. 2. The benefits of publishing with F1000Research: Your article is published within days, with no editorial bias • You can publish traditional articles, null/negative results, case reports, data notes and more • The peer review process is transparent and collaborative • Your article is indexed in PubMed after passing peer review • Dedicated customer support at every stage • For pre-submission enquiries, contact research@f1000.com Page 10 of 10	38,723
dumas-03779317-Med_generale_2022_Vanderstraeten.txt_1	French-Science-Pile	Open Science	Various open science	2,022	Analyse de la pratique gynécologique des médecins généralistes du département des Landes : une étude quantitative descriptive. Médecine humaine et pathologie. 2022. ⟨dumas-03779317⟩	None	French	Spoken	7,060	11,233	Analyse de la pratique gynécologique des médecins généralistes du département des Landes : une étude quantitative descriptive Mathilde Vanderstraeten 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 recherche français trangers des laboratoires publics ou privés. Université de Bordeaux U.F.R DES SCIENCES MEDICALES Année 2022 N°73 Thèse pour l’obtention du DIPLOME d’ETAT de DOCTEUR EN MEDECINE Présentée et soutenue publiquement le 30 juin 2022 par Mathilde VANDERSTRAETEN Née le 17 décembre 1990 à Lille Analyse de la pratique gynécologique des médecins généralistes du département des Landes : une étude quantitative descriptive. Sous la direction du Docteur Élisa LAMAZOU Jury Professeur Bernard GAY ............................... ................................................................Président Professeur Jean-Philippe JOSEPH.................................................................Rapporteur et Juge Docteur Marco ROMERO....................................................................................................Juge Docteur Véronique CHEMINADE-DARMAILLACQ.........................................................Juge Docteur Élisa LAMAZOU................................................................................Directrice et Juge 2 REMERCIEMENTS A Monsieur le Professeur Bernard GAY, Professeur émérite du département de médecine générale de Bordeaux. Vous me faites l’honneur de présider mon jury, merci d’avoir accepté avec un tel enthousiasme. Je tiens à vous témoigner mon respect et ma reconnaissance. A Monsieur le Professeur Jean-Philippe Joseph, Professeur de médecine générale, directeur du département de médecine générale de Bordeaux et médecin généraliste à Bordeaux. Je vous remercie d’avoir accepté d’être le rapporteur de ma thèse et de me faire l’honneur de juger ce travail, pour le temps consacré à la lecture de cette thèse et les remarques judicieuses que vous m’avez indiquées. Soyez assuré de mon profond respect et de ma sincère gratitude. A Madame le Docteur Véronique Cheminade-Darmaillacq, Médecin généraliste à Amou pratiquant uniquement la gynécologie, détentrice d’un DU de gynécologie et d’un DIU d’échographie générale dont les modules gynécologie et obstétrique. Merci d’avoir accepté d’intégrer mon jury de thèse et d’évaluer mon travail. Soyez assurée de mon profond respect et de ma reconnaissance. A Monsieur le Docteur Marco ROMERO, Maître de conférences associé du département de médecine générale de Bordeaux et médecin généraliste à Samadet. Merci d’avoir accepté d’intégrer mon jury de thèse et d’évaluer mon travail. Soyez assuré de mon profond respect et de ma reconnaissance. A Madame le Docteur Élisa Lamazou, Médecin urgentiste au CH de Dax. Je te remercie d’avoir accepté de diriger ma thèse. Après tout ce temps et toutes ces péripéties, on y est! Pour toutes ces journées au SAU en ta compagnie, même si tu nous abandonnes beaucoup trop souvent en sorties SMUR! A tous les médecins généralistes ayant accepté de participer à cette étude, Je vous remercie de m’avoir accordé un peu de votre temps si précieux pour répondre à mon questionnaire. A mes parents, Papa, merci de nous avoir transmis ta passion pour ce métier depuis notre enfance en nous emmenant faire tes visites à moto, en nous racontant toutes ces anecdotes. Pour tous ces « Alors tu es dans les temps? » pendant mes révisions qui pouvaient me stresser mais me motivaient d’autant plus. Maman, merci de t’être pliée en quatre pour nous quand on rentrait le weekend, pour tous ces petits messages d’encouragements (même les messages vocaux juste pour dire que tu avais appelé) et les petites attentions du dimanche soir pour s’assurer qu’on ne manque de rien la semaine. Grâce à vous, tout a été plus facile. Je vous aime. A Poupette, Merci pour ton soutien inconditionnel, pour les semaines de révisions passées à Biache et bien sûr pour les petites patates! Je t’aime. A Camille, ma sœur, ma confidente, Merci d’être là en toutes circonstances, pour tous tes conseils, pour tous ces fous rires, pour ces heures passées à te regarder jouer... Je serai toujours là pour toi. Je t’aime. A Simon, Merci pour ton soutien sans faille et pour ton implication immense dans ce travail mais aussi de tous les jours. De tout cœur, merci pour tout. A Fati et Mosti, pour tous ces moments précieux qu’on a partagés, si importants à mes yeux et que je n’oublierai jamais. On continuera de danser ensemble sous n’importe quel temps, même la pluie! A Imane et Loïc (et Maël), pour toutes ces soirées, ces weekends et ces surprises si bien organisés par la Mama. 4 Rhizlan et Mohammed, pour toute la joie que vous m’apportez et de nous accueillir Camille et moi à bras ouverts, comme vos filles. A toute la famille du Maroc, ma deuxième famille. A Hélène et Azzedine (et Axel), notre amitié m’est chère et j’ai hâte de rencontrer votre petite merveille. A mes amis d’internat, Marion et Pierrick, Margaux, Thibaut et Baptiste, Philippine, Célia et Thomas, Stéphanie, Julie, Quentin, Adri en, Marie, Isabelle, Jérémy et bien d’autres. Merci de m’avoir accompagnée dans cette aventure et je l’espère dans les prochaines à venir. A Alexia et Christophe (et Naomi), pour les apéros, les soirées Halloween, les pauses sur les berges de l’Adour (!), etc... Il faut qu’on fasse tout ça plus souvent! A Julie, ma coloc par intermittence qui va bientôt voler de ses propres ailes! Merci pour ton écoute, tes conseils et ces petites soirées qui font tellement de bien entre deux jours de boulot. A mes collègues, ce n’est pas toujours évident aux urgences mais en votre compagnie tout est plus facile. Je suis ravie d’avoir une équipe comme la nôtre. A Thomas, alias Clark (oui c’est très vieux, comme nous) Pour m’avoir remis dans le droit chemin plus d’une fois durant toutes ces années! Malgré la distance, rien ne change et merci d’être là. A la bande de fous de Pau, Nico La Nique, Tim La Boule et Lord Paulo de L.A, membres de la Rocket Company, pour ces soirées sardines, ces soirées Dudu (qu’on s’en souvienne ou non) et toutes les autres. Vous êtes fous mais qu’est-ce qu’on se marre. A toute ma famille, malgré la distance je pense à vous. A Calie et Roger. 5 A Thibault, mon frère ... AUX ................ 9 RESUME.................................................................................................................................. 11 ABSTRACT............................................................................................................................. 13 GLOSSAIRE............................................................................................................................ 15 INTRODUCTION.................................................................................................................... 16 I. Rôle du médecin généraliste........................................................................................ 16 II. Démographie médicale................................................................................................. 16 1. Démographie des médecins généralistes.................. ................ ................................ 16 2. Démographie des gynécol ogue s ............................................................................... 17 III. Place du médecin généraliste dans la prise en charge gynécologique des patientes 17 MATERIELS ET METHODES............................................................................................... 19 I. Matériels....................................................................................................................... 19 1. Population................................................................................................................. 19 2. Critères d’inclusion.................................................................................................. 19 II. Méthodes...................................................................................................................... 19 1. Type d’étude............................................................................................................. 19 2. Recrutement............................................................................................................. 20 3. Recueil des données................................................................................................. 20 a. Déroulement de l’enquête.................................................................................... 20 b. Questionnaire....................................................................................................... 21 4. Analyse des données................................................................................................ 21 III. Étude de la littérature............................................................................................... 22 RESULTATS........................................................................................................................... 23 I. Caractéristiques des Médecins Généralistes................................................................ 23 1. Population étudiée.................................................................................................... 23 2. Répartition des médecins selon le sexe et l’âge....................................................... 25 3. Mode d’exercice et proximité avec un gyn écologue ................................................ 26 4. Nombre de consultations de médecine général e...................................................... 27 5. Formation initiale et formation com plémentaire ...................................................... 28 II. Caractéristiques de la pratique gynécologique............................................................. 29 1. Pratique gynécologique globale............................................................................... 29 2. Pratique gynécologique selon le sexe....................................................................... 33 3. Pratique gynécologique selon l’âge.......................................................................... 34 7 4. Pratique gynécologique selon la proximité avec un gynécologue........................... 36 5. Pratique gynécologique selon la formation.............................................................. 37 6. Freins à la pratique gynécologique........................................................................... 39 7. Motivations à la pratique gynécologique................................................................. 43 8. Projection sur la demande........................................................................................ 47 DISCUSSION.......................................................................................................................... 49 I. Analyse des résultats.................................................................................................... 49 1. Données démographiques........................ ................................................................ 49 2. Étude des résultats .................................................................................................... 49 II. Critiques méthodologiques........................................................................................... 61 1. Biais de sélection...................................................................................................... 61 2. Biais d’ interprétation ................................................................................................ 62 3. Biais de déclaration .................................................................................................. 62 III. Atouts de l’étude...................................................................................................... 63 CONCLUSION........................................................................................................................ 64 BIBLIOGRAPHIE................................................................................................................... 66 ANNEXES............................................................................................................................... 71 SERMENT D’HIPPOCRATE................................................................................................. Contexte : Le suivi gynécologique est essentiellement assuré par les gynécologues, bien qu’il soit accessible aux compétences du médecin généraliste. Les projections de démographie médicale annoncent une pénurie de gynécologues. Dans le département des Landes, cette diminution du nombre de gynécologues depuis 2010 atteint 63,6% pour les gynécologues obstétriciens et 25% pour les gynécologues médicaux. Ce constat pourrait amener les médecins généralistes à devoir accroitre leur activité gynécologique. L’objectif principal de cette étude est de réaliser une évaluation de la pratique de la gynécologie des médecins généralistes des Landes. Méthode : Étude épidémiologique descriptive par questionnaire adressé aux médecins généralistes installées dans le département des Landes. Les médecins remplaçants, salariés et exerçant une « activité particulière » (angiologie, thermalisme...) ont été exclus. Le questionnaire a été envoyé par courrier électronique entre octobre 2019 et juillet 2020 et comprenait trois parties : caractéristiques des répondants, état des lieux de leur activité globale et gynécologique et évaluation des motivations et frein à leur pratique de la gynécologie. Résultats : Sur les 283 questionnaires envoyés, 145 réponses ont été obtenues (51,2%). Les femmes réalisent davantage les actes courants de gynécologie que les hommes. Quelque soient leurs caractéristiques, les médecins déclarent réaliser peu de gestes techniques : 3% posent un DIU et 2% un implant). L’âge du médecin et la distance avec le gynécologue ne modifiaient pas la pratique des médecins. Les médecins s’estimant mieux formés ou ayant réalisé une formation complémentaire pratiquaient davantage la gynécologie mais 70,3% des médecins estimaient leur formation initiale comme insuffisante. Les principaux freins rencontrés étaient la proximité avec un gynécologue (4 et le manque de demande des patientes (35,9%). La majorité des répondants (63%) accepteraient de faire davantage de gynécologie si la demande était plus forte. 11 Conclusion : Une meilleure information de la patiente sur les compétences du médecin généraliste, une répartition plus précise des rôles de chacun des acteurs de la prise en charge gynécologique et une formation initiale plus solide amélioreraient la pratique du suivi gynécologique par les médecins généralistes. 12 ABSTRACT Title: Analysis of the gynecological practice of general practitioners in the Landes department: a quantitative descriptive study. Context : The regular gynecological follow-up is essentially performed by gynecologists, although it is accessible through the expertise of general practitioners. Medical demographic projections predict a decline in the number of gynecologists. Since 2010 in the Landes department, this decrease in the number of gynecologists reached 63.6% for obstetrician gynecologists and 25% for medical gynecologists. This observation could lead general practitioners to increase their gynecological activity. The main objective of this study is to evaluate the practice of gynecology by general practitioners in the Landes. Method: Descriptive and quantitative epidemiological study by questionnaire sent to general practitioners in the Landes department between October 2019 and July 2020. Substitute doctors, salaried doctors, and doctors practising a "particular activity" (angiology, thermalism, etc.) were excluded. The questionnaire was sent by e-mail and included three parts: characteristics of the respondents, assessment of their general and gynecological activity, and evaluation of the motivations and obstacles to their practice of gynecology. Results: Of the 283 questionnaires sent to the 313 doctors included, 145 responses were received. This represents a participation rate of 51.2%. Female general practitioners performed more routine gynecological procedures than male doctors. Regardless of their characteristics, the doctors declared they performed few technical procedures (97% for the IUD and 98% for the contraceptive implant). Age and distance from the gynecologist did not affect doctors’ practice. 70.3% of the doctors surveyed considered their initial training to be sufficient. Doctors who felt they were better trained or had completed supplementary training practised more gynecology. The main obstacles encountered were proximity to a gynecologist (41%) and lack of demand from patients (35.9%). Most respondents (63%) would accept to do more gynecology if there was more demand. 13 Conclusion: Better information for the patient on the GP's competences, a more precise distribution of the roles of each actor in gynecological care and the interest of more intensive training would improve the quality of women's follow-up. 14 GLOSSAIRE CDOM 40 = Conseil Départemental de l’Ordre des Médecins des Landes CNOM = Conseil National de l’Ordre des Médecins CPAM = Caisse Primaire d’Assurance Maladie DIU = Dispositif intra-utérin DU = Diplôme universitaire FC = Formation complémentaire FCV = Frottis cervico-vaginaux FI = Formation initiale FMC = Formation médicale continue GM = Gynécologues médicaux GO = Gynécologues obstétricaux HPV = Human Papilloma Virus (Papillomavirus humain) IST = Infection sexuellement transmissible Rég. = Régulièrement TV = Toucher vaginal URPS ML-NA = Union Régionale des Professionnels de Santé – Médecins Libéraux de Nouvelle-Aquitaine 15 INTRODUCTION I. Rôle du médecin généraliste Le médecin généraliste joue un rôle primordial dans la prise en charge globale du patient. Comme le précise l’Assurance Maladie, le médecin traitant assure les soins de premier recours habituels et de prévention. Il participe activement à la bonne utilisation du système de soins par ses patients. Il met également en place un suivi médical personnalisé et dirige son patient vers un médecin spécialiste en cas de nécessité (1). En 2002, la Société Européenne de Médecine générale – Médecine de famille (WONCA Europe) a défini les caractéristiques essentielles de la médecine générale. « Le médecin généraliste est le premier contact avec le système de soins, permettant un accès ouvert et non limité aux usagers, prenant en compte tous les problèmes de santé, indépendamment de l’âge, du sexe, ou de toutes autres caractéristiques de la personne concernée » (2). Une femme consulte son médecin généraliste en moyenne 3,6 fois par an pour un motif d’ordre gynécologique et cette fréquence augmente avec l’âge de la patiente (3). Alors qu’en 1997, 60% des femmes consultaient régulièrement et spontanément leur gynécologue médical, en 2012 elles n’étaient plus que 25% à pouvoir le faire, faute d’effectifs nécessaires (4). Cela les pousse à consulter davantage leur médecin traitant pour ce motif. II. Démographie médicale Le CNOM a publié un atlas de démographique médicale (5). 1. Démographie des médecins généralistes En 2018, on recensait en France 87 801 médecins généralistes en activité régulière (tous mode d’exercice) soit une diminution de 7% depuis 2010 dont 0,4% entre 2017 et 2018. Cette diminution atteint même 1,4% dans le département des Landes entre ces mêmes dates (Annexe 1). 16 Cette tendance à la baisse à une forte probabilité de se confirmer jusqu’en 2025, ce qui s’explique par la pyramide des âges qui illustre le vieillissement de la profession (âge moyen de 50,6 ans et 24% de plus de 60 ans), les départs en retraites et le recul de l’âge de la première installation (Annexe 2 et 3). Selon l’URPSML-NA, le département des Landes comptabilise une densité de 548,1 médecins généralistes pour 100 000 femmes en âge de procréer (de 15 à 49 ans) (Annexe 4). 2. Démographie des gynécologues Le rapport du CNOM de 2018 recensait en France 2230 gynécologues dont 1105 gynécologues obstétriciens (GO) et 1125 gynécologues médicaux (GM). Alors qu’en 2010 on comptait 3525 gynécologues dont 1886 GO et 1639 GM. Dans les Landes, cette diminution du nombre de gynécologues depuis 2010 atteint 63,6% pour les GO et 25% pour les GM (7) (Annexe 5). Selon l’URPSML-NA, le département des Landes comptabilise une densité de 12,7 GO et 17,8 GM pour 100 000 femmes en âge de procréer (15-49ans) (Annexe 4). Cela représente une diminution plus rapide que celle des médecins généralistes. De plus, la pyramide des âges des gynécologues illustre parfaitement cette pénurie à venir (Annexe 6). En effet, près de la moitié des gynécologues des Landes sont proches de la retraite avec une proportion de plus de 60 ans chiffrée à 50% (7). Cette tendance est expliquée par la réforme de l’internat de 1984. En effet la spécialité gynécologie médicale a été supprimée du cursus universitaire pendant environ vingt ans. Cette diminution d’effectifs de gynécologues a ainsi soulevé un problème potentiel de santé publique engendrant une éventuelle incapacité à assurer un suivi gynécologique régulier. Et ce ne qu’à partir de 2003 qu’une augmentation régulière du nombre de postes d’internes en gynécologie médicale a commencé à être observée. Ces postes ont même plus que doublé en trois ans passant de 30 en 2012 à 68 en 2015 (8). III. Place du médecin généraliste dans la prise en charge gynécologique des patientes Devant cette pénurie annoncée de gynécologues et l’augmentation démographique de la population féminine (accroissement de la population et allongement de l'espérance de vie) (9), le Conseil National des Gynécologues Obstétriciens Français (CNGOF) confirme que « les 17 spécialistes de la gynécologie obstétrique n’ont pas pour vocation de voir toutes les femmes pour les problèmes de contraception, les examens systématiques et de dépistage, les thérapeutiques les plus courantes ou le traitement substitutif de la ménopause »(10). En effet, celui-ci souligne la nécessité d’une collaboration avec les médecins généralistes afin que ces derniers soient en première ligne en ce qui concerne les actes courants de gynécologie. Tous ces éléments montrent que le médecin généraliste joue un rôle important dans le suivi gynécologique de ses patientes et qu’il va devoir répondre à une demande potentiellement croissante en gynécologie. L’objectif principal de cette étude est de réaliser une évaluation de la pratique de la gynécologie des médecins généralistes des Landes. L’objectif secondaire de l’étude est de cibler leurs motivations ou freins potentiels à cette pratique afin de leur en faciliter l’exercice. 18 MATERIELS ET METHODES I. Matériels 1. Population La population cible était celle des médecins généralistes libéraux exerçant dans le département des Landes en 2019. 2. Critères d’inclusion Les critères d’inclusion étaient : - Médecin généraliste - Activité libérale - Installé dans le département des Landes (40). Les critères d’exclusion étaient : - Mode d’exercice majoritairement salarié/hospitalier - Médecins à « exercice particulier » (médecine thermale, nutrition, acupuncture, angiologie...) - Remplacement. II. Méthodes Type d’étude Il s’agit d’une étude épidémiologique quantitative avec analyse descriptive des données recueillies par questionnaire. Recrutement Pour des raisons de protection des données personnelles, l’Union Régionale des Professionnels de Santé – Médecins Libéraux de Nouvelle-Aquitaine (URPS-ML-NA) et le Conseil Départemental de l’Ordre des Médecins des Landes (CDOM 40) n’ont pas divulgué la liste des médecins généralistes car ils ne pouvaient pas en extraire les coordonnées téléphoniques ou électroniques des médecins. La liste de ces médecins a donc été obtenue via le site du Conseil National de l’Ordre des Médecins (CNOM) et via le site Ameli.fr de la Caisse Primaire d’Assurance Maladie (CPAM) des Landes. La récupération des données téléphoniques a été faite par recherches internet, essentiellement sur le site des Pages Jaunes. Chaque cabinet a été appelé afin d’obtenir les adresses électroniques des médecins en fonction des critères d’inclusion. 3. Recueil des données a. Déroulement de l’enquête Un message électronique envoyé à chaque médecin de la liste comprenait un texte explicatif de l’objet de ce message, une courte description du sujet de la thèse et le lien pour remplir le questionnaire (Annexe 7). Le recueil des données s’est déroulé d’octobre 2019 à juillet 2020. Plusieurs relances ont été nécessaires afin d’obtenir un échantillon représentatif. La réponse au questionnaire étant anonyme, les mails de relance ont été envoyés à tous les médecins en leur précisant de « ne pas y répondre si déjà fait ». 20 b. Questionnaire Avec ma directrice de thèse le Dr LAMAZOU Élisa, nous avons opté pour un questionnaire à réponses fermées, anonyme et court afin de récolter le maximum de réponses possibles (Annexe 8). Celui-ci a été élaboré via la plateforme Google Forms et comprenait : première partie descriptive des caractéristiques du médecin : âge, sexe, mode d’exercice, formation en gynécologie et distance d’un gynécologue (hospitalier ou libéral). - Une deuxième partie sur la quantification du nombre de consultation de médecine générale tous motifs confondus et des actes de gynécologie réalisés. Pour que le remplissage du questionnaire soit plus aisé pour les médecins, le nombre de consultations de médecine générale a été demandé par semaine et le nombre d’actes de gynécologie demandé par mois. - Une dernière partie sur l’évaluation des difficultés rencontrées freinant leur pratique de la gynécologie. La liste de ces difficultés a été établie selon les principaux freins évoqués dans la littérature. 4. Analyse des données Les données ont été extraites de manière automatisée par la plateforme Google Forms sous forme d’un tableur Excel. Les données qualitatives ont été exprimées en termes d’effectifs et de pourcentages. Les différents tests (χ 2, test exact de Fisher) de comparaison de données (pourcentages) ont été effectués sur le logiciel R. Un test du χ 2 d’homogénéité a été utilisé lorsque la règle de Cochran était respectée (au moins 80% des valeurs théoriques étaient au moins égales à 5, et que toutes les valeurs théoriques étaient supérieures à 1). Dans le cas contraire, le test exact de Fisher a été choisi. III. Étude de la littérature Les bases de données les plus pertinentes concernant la thématique ont été interrogées. Les équations de recherches ont été adaptées au thesaurus de chacune d’entre elles, avec pour mots-clés « gynécologie » OU « suivi gynécologique » OU « pratique gynécologique » ET « médecine générale » OU « médecin généraliste ». La plupart des références bibliographiques ont été trouvées sur la base de données Sudoc, ainsi que par des articles parus dans la revue Exercer. Les résultats ont été enrichis de sources trouvées indirectement, notamment par le biais des bibliographies des thèses similaires consultées. La thèse de H. Guyomard a été très utile puisqu’il s’agit d’une revue de la littérature du suivi gynécologique en médecine générale (7). 22 RESULTATS I. Caractéristiques des Médecins Généralistes 1. Population étudiée Selon le site du CNOM et Ameli.fr de la CPAM des Landes, 424 médecins exercent dans le département des Landes en 2019. Sur ces 424 médecins généralistes, 60 ont été exclus car ils ne répondaient pas aux critères d’inclusion et 51 n’ont jamais répondu à l’appel téléphonique après trois tentatives. Parmi les 313 médecins finalement retenus pour participer à cette étude, 25 ont refusé d’y participer et 5 avaient une adresse électronique erronée. Le questionnaire a donc été envoyé à 283 médecins et 145 réponses ont été obtenues, soit un taux de participation au questionnaire de 51,2%. 23 Figure 1: Flow Chart 24 2. Répartition des médecins selon le sexe et l’âge Sexe Effectifs (%) Femmes 74 (51) Hommes 71 (49) Total 145 Tableau 1 : Répartition des médecins selon le sexe La population est donc représentée par 145 médecins dont 74 femmes et 71 hommes, soit un sexe ratio H/F de 0,96. Effectifs (%) Tranches d’âge Total Hommes Femmes (n = 71) (n = 74) 25 – 40 ans 13 (18,4) 28 (37,8) 41 (28,3) 41 – 59 ans 29 (40,8) 33 (44,6) 62 (42,7) > 60 ans 29 (40,8) 13 (17,6) 42 (29) 145 Tableau 2 : Répartition des médecins selon la tranche d’âge et le sexe La tranche d’âge la plus représentée est la tranche des 41-59 ans (42,7%). Les médecins généralistes de sexe féminin sont significativ plus jeunes que leurs homologues masculins (p < 0.01, test du Chi2 de Pearson). Mode d’exercice et proximité avec un gynécologue Mode d’exercice Effectifs (%) Rural 45 (31) Semi-rural 70 (48,3) Urbain 30 (20,7) Total 145 Tableau 3 : Répartition des médecins selon leur mode d’exercice Les modes d’exercice ou secteurs d’activité sont répartis en 3 catégories : rural, semi-rural et urbain. La pratique en milieu semi-rural est le mode d’exercice majoritairement représenté puisqu’il a récolté près de la moitié des réponses (48,3%), soit 70 médecins. Effectifs (%) Temps de Urbain Semi-rural Rural Total trajet n = 29 n = 70 n = 45 < 15 min 29 (100) 40 (57,1) 4 (8,9) 73 16 – 30 min 0 (0) 21 (30) 24 (53,3) 45 31 min – 1h 0 (0) 9 (12,9) 16 (35,6) 25 >1h 0 (0) 0 (0) 1 (2,2) 1 144 Tableau 4 : Répartition des médecins selon leur mode d’exercice et leur temps de trajet pour se rendre chez le gynécologue le plus proche La moitié des médecins a répondu être en proximité immédiate d’un gynécologue : 73 d’entre eux (soit 50,7%) sont à moins de 15min de celui-ci. Il existe une différence significative du temps de trajet pour se rendre chez le gynécologue le plus proche selon le lieu d’exercice du médecin généraliste (p < 0.001, test exact de Fisher) ; 26 les médecins exerçant en milieu rural ont plus difficilement accès à une consultation avec un gynécologue que leurs confrères exerçant en milieu urbain par leur éloignement avec ce spécialiste. Le nombre total de réponses à cette question est de 144. Une d’entre elles a été exclue pour deux propositions contradictoires cochées (<15min et 31min - 1h). 4. Nombre de consultations de médecine générale Nombres de consultations par semaine Effectifs (%) < 50 / semaine 19 (13,1) 51 – 100 / semaine 41 (28,3) 101 – 150 / semaine 69 (47,6) > 150 / semaine 16 (11) Tableau 5 : Répartition des médecins en fonction du nombre hebdomadaire de consultations de médecine générale réalisées Effectifs (%) Sexe Total < 50 / sem 51 – 100 / sem 101 – 150 / sem > 150 / sem n = 19 n = 41 n = 69 n = 16 Hommes 11 (57,9) 14 (34,1) 35 (50,7) 11 (68,75) 71 Femmes 8 (42,1) 27 (65,9) 34 (49,3) 5 (31,25) 74 145 Table au 6 : Ré partition des médecins selon le sex e et le nombre de consultations hebdomadaires de médecine générale Les hommes semblent réaliser globalement plus de consultations de médecine générale par semaine que les femmes (p = 0.078, Test du Chi2 de Pearson). 27 Effectifs (%) Nombre de Total 25 – 40 ans 41 – 59 ans > 60 ans semaine n = 41 n = 62 n = 42 < 50 / semaine 4 (9,8) 9 (14,5) 6 (14,3) 19 51 – 100 / semaine 16 (39) 18 (29) 7 (16,7) 41 101 – 150 / semaine 18 (43,9) 28 (45,2) 23 (54,7) 69 > 150 / semaine 3 (7,3) 7 (11,3) 6 (14,3) 16 consultations par 145 Tableau 7 : Répartition des médecins selon l’âge et le nombre de consultations hebdomadaires de médecine générale L’âge du praticien n’exerce aucune influence sur le nombre de consultations hebdomadaires (p = 0.45, Test de Fisher). 5. Formation initiale et formation complémentaire Formation initiale suffisante? Effectifs (%) Oui 43 (29,7) Non 102 (70,3) Total 145 Tableau 8 : Répartition des médecins en fonction de l’avis sur leur formation initiale en gynécologie 28 Réalisation Effectifs (%) formation FI suffisante FI non suffisante Total complémentaire? n = 43 n = 102 Oui 25 (58,1) 32 (31,4) 57 Non 18 (41,9) 70 (68,6) 88 145 Tableau 9 : Répartition des médecins en fonction de leur avis sur leur formation initiale et de leur réalisation de formation complémentaire Sur les 102 médecins estimant leur formation initiale (FI) comme non suffisante, 70 n’ont pas réalisé de formation complémentaire (FC) ou de stage en gynécologie (68,6%). Les praticiens qui considèrent leur FI comme suffisante réalisent significativement plus de FC que leurs confrères l’estimant comme non suffisante (p = 0.0047, Test du Chi2 de Pearson avec correction de Yates). Sur les 57 médecins ayant réalisé une FC en gynécologie, 35 étaient des femmes (61,4%). Ces résultats montrent que les femmes semblent réaliser plus de FC que les hommes, sans être toutefois significatifs (p = 0.066, Test du Chi2 de Pearson avec correction de Yates). II. Caractéristiques de la pratique gynécologique 1. Pratique gynécologique globale Les différents actes gynécologiques étudiés sont : - L’examen gynécologique général : sénologique, toucher vaginal (TV)... - La prescription ou renouvellement de contraception - Le bilan d’infection(s) sexuellement transmissible(s) (IST) - La vaccination contre le papillomavirus (HPV) - Le suivi de la ménopause - Le suivi de grossesse 29 - La réalisation du frottis cervico-vaginal (FCV) - La pose/le retrait d’implant contraceptif - La pose/le retrait de dispositif intra-utérin (DIU) Figure 2 : Répartition des médecins en fonction de la fréquence mensuelle de réalisation des différents actes de gynécologie Voici une vue d’ensemble de la pratique gynécologique des médecins ayant répondu au questionnaire. Ce graphique est intéressant car il permet de visualiser en un seul coup d’œil la pratique des médecins en fonction des actes. La fréquence de réalisation des différents actes de gynécologie est répartie en quatre catégories en fonction du nombre de consultations mensuelles : - Pas : 0 / mois - Peu : 1 à 5 / mois - Régulièrement (Rég.) : 5 à 10 / mois - Beaucoup : > 10 / mois. 30 Actes réalisés Examen gynécologique Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Réalisation des FCV Pose / retrait implant Fréquence Pas / Peu 119 (82) Régulièrement / Beaucoup 26 (18) Pas / Peu 76 (52) Régulièrement / Beaucoup 69 (48) Pas / Peu 107 (74) Régulièrement / Beaucoup 38 (26) Pas / Peu 113 (78) Régulièrement / Beaucoup 32 (22) Pas / Peu 117 (81) Régulièrement / Beaucoup 28 (19) Pas / Peu 141 (98) Régulièrement / Beaucoup 3 (2) * Pas / Peu 124 (86) Régulièrement / Beaucoup 21 (14) Pas / Peu 142 (98) Régulièrement / Beaucoup Pose / retrait DIU Effectifs (%) Pas / Peu Régulièrement / Beaucoup 3 (2) 141 (97) 4 (3) * A cette question, total de réponses de 144 et non 145 comme les autres résultats puisqu’une réponse a été exclue car erronée. Tableau 10 : Répartition des actes gynécologiques selon leur fréquence de réalisation par les médecins de l’étude On peut voir que l’activité gynécologique des médecins généralistes est très variable en fonction du motif de consultation (figure 2) (tableau 10) (Annexe 9). Si près de la moitié des médecins (48%) déclare réaliser « régulièrement » ou « beaucoup » les consultations abordant le sujet de la contraception, leur pratique est plus hésitante concernant les autres motifs gynécologiques. Ils sont une majorité à avoir répondu « peu » réaliser le bilan IST, le suivi de ménopause et la vaccination HPV. 31 Pour les autres motifs de consultation ils sont une majorité déclarant ne pas en faire. Si les réponses sont unanimes sur l’absence totale de réalisation des gestes techniques de la contraception (85% pour l’implant et 90% pour le DIU), on peut tout même nuancer les résultats pour l’examen gynécologique, le suivi de grossesse et le FCV. Si 46% des médecins ne réalisent pas d’examen gynécologique, ils sont malgré tout 36% à en réaliser « peu » et 18% à en réaliser « régulièrement » ou « beaucoup ». Concernant le suivi de grossesse, 60% n’en réalisent pas du tout mais 38% en réalisent « peu ». De même pour le FCV, 61% n’en réalisent pas du tout mais 25% en réalisent « peu » et 14% le font régulièrement ou beaucoup. 32 2. Pratique gynécologique selon le sexe Effectifs (%) Actes réalisés Examen gynéco Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Fréquence Hommes Femmes n = 71 n = 74 Pas / Peu 69 (97) 50 (68) Régulièrement / Beaucoup 2 (3) 24 (32) Pas / Peu 46 (65) 30 (41) Régulièrement / Beaucoup 25 (35) 44 (59) Pas / Peu 59 (83) 48 (65) Régulièrement / Beaucoup 12 (17) 26 (35) Pas / Peu 58 (82) 55 (74) Régulièrement / Beaucoup 13 (18) 19 (26) Pas / Peu 60 (85) 57 (77) Régulièrement / Beaucoup 11 (15) 17 (23) Pas / Peu 68 (97) 73 (99) Régulièrement / Beaucoup 2 (3) 1 (1) 70 (99) 54 (73) 1 (1) 20 (27) 71 (100) 71 (96) 0 (0) 3 (4) 71 (100) 70 (95) 0 (0) 4 (5) Réalisation FCV Pas / Peu Régulièrement / Beaucoup Pose/retrait Pas / Peu implant Régulièrement / Beaucoup Pose/retrait DIU Pas / Peu Régulièrement / Beaucoup p* < 0.01 < 0.01 0.013 0.29 0.25 0.62F <0.01 0.25F 0.12F * Test du Chi2 de Pearson ou F Test exact de Fisher A cette question, total de réponses de 70 et non 71 comme les autres résultats concernant les hommes puisqu’une réponse a été exclue car erronée. Tableau 11 : Répartition des actes gynécologiques selon leur fréquence de réalisation et le sexe des médecins de l’étude Les femmes réalisent significativement plus d’examen gynécologique, de prescription de contraception et de FCV que leurs confrères masculins (p <0.01). Elles réalisent également significativement plus de bilan d’IST que les hommes (p = 0.013). 33 En revanche il n’y a pas de différence significative selon le sexe du praticien en ce qui concerne le suivi de ménopause (p = 0.25), le suivi de grossesse (p=0.62), la vaccination HPV (p = 0.29) et la pose ou retrait d’implant (p = 0.25) et de DIU (p = 0.12). 3. Pratique gynécologique selon l’âge Effectifs (%) Actes réalisés Fréquence 25 – 40 ans 41 – 59 ans > 60 ans n = 41 n = 62 n = 42 Pas / Peu 32 (78) 54 (87) 33 (79) Rég. / Beaucoup 9 (22) 8 (13) 9 (21) Pas / Peu 24 (59) 33 (53) 19 (45) Rég. / Beaucoup 17 (41) 29 (47) 23 (55) Pas / Peu 29 (71) 46 (74) 32 (76) Rég. / Beaucoup 12 (29) 16 (26) 10 (24) Pas / Peu 31 (76) 52 (84) 30 (71) Rég. / Beaucoup 10 (24) 10 (16) 12 (29) Pas / Peu 36 (88) 49 (79) 32 (76) Rég. / Beaucoup 5 (12) 13 (21) 10 (24) Pas / Peu 38 (95) 61 (98) 42 (100) Rég. / Beaucoup 2 (5) ** 1 (2) 0 (0) 34 (83) 56 (90) 34 (81) Rég. / Beaucoup 7 (17) 6 (10) 8 (19) Pose/retrait Pas / Peu 40 (98) 60 (97) 42 (100) implant Rég. / Beaucoup 1 (2) 2 (3) 0 (0) Pose/retrait DIU Pas / Peu 39 (95) 60 (97) 42 (100) Rég. / Beaucoup 2 (5) 2 (3) 0 (0) Examen gynéco Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Réalisation FCV Pas / Peu p* 0.39 0.47 0.85 0.30 0.37 0.36F 0.35 0.63F 0.39F * Test du Chi2 de Pearson ou F Test exact de Fisher A cette question « suivi de grossesse », total de réponses de 40 et non 41 comme les autres résultats concernant les 25 - 40 ans puisqu’une réponse a été exclue car erronée. Tableau 12 : Répartition des actes gynécologiques selon leur fréquence de réalisation et l’âge des médecins de l’étude 34 Les résultats montrent que l’âge du praticien n’influe pas sur la réalisation des différents actes de gynécologie que ce soit en termes de réalisation de consultation simple ou d’actes techniques. 35 4. Pratique gynécologique selon la proximité avec un gynécologue Effectifs (%) Actes réalisés Proche Éloigné (< 15 min) (> 15min) n = 73 n = 71 Pas / Peu 60 (82) 59 (83) 119 Rég. / Beaucoup 13 (18) 12 (17) 25 Pas / Peu 41 (56) 35 (49) 75 Rég. / Beaucoup 32 (44) 36 (51) 68 Pas / Peu 53 (73) 54 (76) 107 Rég. / Beaucoup 20 (27) 17 (24) 37 Pas / Peu 61 (84) 51 (72) 112 Rég. / Beaucoup 12 (16) 20 (28) 32 Pas / Peu 58 (79) 58 (82) 116 Rég. / Beaucoup 15 (21) 13 (18) 28 Pas / Peu 71 (99) * 69 (97) 140 Rég. / Beaucoup 1 (1) * 2 (3) 3 63 (86) 61 (86) 124 Rég. / Beaucoup 10 (14) 10 (14) 20 Pose/retrait Pas / Peu 72 (99) 69 (97) 141 implant Rég. / Beaucoup 1 (1) 2 (3) 3 Pose/retrait DIU Pas / Peu 71 (97) 69 (97) 140 Rég. / Beaucoup 2 (3) 2 (3) 4 Fréquence Examen gynéco Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Réalisation FCV Pas / Peu Total p* 0.89 0.41 0.64 0.09 0.73 0.62F 0.95 0.62F 1F * Test du Chi2 de Pearson ou F Test exact de Fisher Un médecin a rendu une réponse erronée concernant son éloignement avec un gynécologue, ses réponses ont donc été exclues portant le nombre total de réponses à 144 et non 145 comme pour les autres items. * A cette question « suivi de grossesse », total de réponses de 72 et non 73 comme les autres résultats concernant la catégorie « Éloigné (> 15min) » puisqu’une réponse a été exclue car erronée. Tableau 13 : Répartition des actes gynécologiques selon leur fréquence de réalisation et la proximité des médecins avec un gynécologue 36 L’éloignement avec un gynécologue n’a montré aucune différence significative dans la pratique des divers actes gynécologiques des médecins généralistes. Pratique gynécologique selon la formation Effectifs (%) Actes réalisés Fréquence FI suffisante FI non suffisante n = 43 n = 102 Pas / Peu 29 (67) 90 (88) Rég. / Beaucoup 14 (33) 12 (12) Pas / Peu 20 (47) 56 (55) Rég. / Beaucoup 23 (53) 46 (45) Pas / Peu 29 (67) 78 (76) Rég. / Beaucoup 14 (33) 24 (24) Pas / Peu 32 (74) 81 (79) Rég. / Beaucoup 11 (26) 21 (21) Pas / Peu 31 (72) 86 (84) Rég. / Beaucoup 12 (28) 16 (16) Pas / Peu 42 (98) 99 (98) Rég. / Beaucoup 1 (2) 2 (2) 33 (77) 91 (89) Rég. / Beaucoup 10 (23) 11 (11) Pose/retrait Pas / Peu 42 (98) 100 (98) implant Rég. / Beaucoup 1 (2) 2 (2) 41 (95) 100 (98) 2 (5) 2 (2) Examen gynéco Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Réalisation FCV Pas / Peu Pose/retrait DIU Pas / Peu Rég. / Beaucoup p* < 0.01 0.36 0.26 0.51 0.08 1F 0.051 1F 0.58F * Test du Chi2 de Pearson ou F Test exact de Fisher A cette question « suivi de grossesse », total de réponses de 101 et non 102 comme les autres résultats concernant la catégorie « FI non suffisante » puisqu’une réponse a été exclue car erronée. Tableau 14 : Répartition des actes gynécologiques selon leur fréquence de réalisation et l’avis des médecins sur leur formation initiale 37 Les praticiens considérant leur FI comme suffisante réalisent significativement plus d’examen gynécologique (p <0.01) que leurs confrères l’estimant comme non suffisante. Nous pouvons étendre cette constatation à la réalisation de FCV puisque les résultats sont tout juste significatifs (p = 0.05). Effectifs (%) Actes réalisés Formation Pas de formation complémentaire complémentaire n = 57 n = 88 Pas / Peu 41 (72) 78 (89) Rég. / Beaucoup 16 (28) 10 (11) Pas / Peu 29 (51) 47 (53) Rég. / Beaucoup 28 (49) 41 (47) Pas / Peu 39 (68) 68 (77) Rég. / Beaucoup 18 (32) 20 (23) Pas / Peu 45 (79) 68 (77) Rég. / Beaucoup 12 (21) 20 (23) Pas / Peu 41 (72) 76 (86) Rég. / Beaucoup 16 (28) 12 (14) Pas / Peu 55 (96) 86 (99) Rég. / Beaucoup 2 (4) 1 (1) ** 42 (74) 82 (93) Rég. / Beaucoup 15 (26) 6 (7) Pose/retrait Pas / Peu 54 (95) 88 (100) implant Rég. / Beaucoup 3 (5) 0 (0) 53 (93) 88 (100) 4 (7) 0 (0) Examen gynéco Contraception Bilan IST Vaccination HPV Suivi ménopause Suivi grossesse Fréquence Réalisation FCV Pas / Peu Pose/retrait DIU Pas / Peu Rég. / Beaucoup p* 0.01 0.77 0.24 0.81 0.03 0.56F < 0.01 0.059F 0.02F * Test du Chi2 de Pearson ou F Test exact de Fisher ** A cette question « suivi de grossesse », n = 87 et non 88 comme les autres résultats concernant la catégorie « Pas de formation complémentaire » puisqu’une réponse a été exclue car erronée. Tableau 15 : Répartition des actes gynécologiques selon leur fréquence de réalisation et la réalisation d’une formation complémentaire par les médecins de l’étude 38 Les médecins généralistes ayant réalisé une FC réalisent significativement plus d’examen gynécologique (p = 0.01) et de suivi de ménopause (p = 0.03) que les médecins n’ayant pas réalisé de FC. Même constatations concernant la réalisation de gestes techniques puisque la différence est significative pour la réalisation de FCV (p < 0.01) et la pose ou retrait de DIU (p = 0.02) et quasiment significative pour la pose ou retrait d’implant (p = 0.059). 6. Rang 1 Freins à la pratique gynécologique Freins Proximité avec autres professionnels Effectifs (%) 59 (41%) 2 Manque de demande 52 (35,9%) 3 Manque de temps 47 (32,4%) 4 Manque de formation 42 (29%) 5 Manque d’intérêt 19 (13,1%) 6 Relation médecin – patientes / Gêne 16 (11%) 7 Manque de matériel 13 (9%) 8 Manque de rémunération 9 (6,2%) Tableau 16 : Classement des freins à la pratique gynécologique selon les médecins généralistes Les quatre freins principaux à la pratique gynécologique rapportés par les médecins généralistes sont la proximité avec un autre professionnel exerçant dans ce domaine (gynécologues, sagefemme), le manque de demande de la part des patientes, le manque de temps et le manque de formation. 2 Tableau 17 : Analyse des différents freins à la pratique gynécologique selon les caractéristiques des médecins généralistes 40 - Proximité avec les autres professionnels Logiquement, la proximité avec un autre professionnel exerçant la gynécologie est un frein significativement plus exprimé par les médecins généralistes ayant leur cabinet à moins de 15min du gynécologue par rapport à leurs confrères qui en sont éloignés (p < 0.01). - Manque de demande Le manque de demande est significativement plus ressenti par les médecins masculins que féminins (p < 0.01). 65,4% des médecins rencontrant ce frein étaient des hommes. - Manque de temps Le manque de temps pour l’exercice de la gynécologie est un frein significativement plus ressenti par les praticiens éloignés d’un gynécologue (p < 0.01) et n’ayant pas réalisé de FC (p = 0.046). Il est à la limite de significativité pour ceux estimant leur FI comme non suffisante (p = 0.055). - Manque de formation Logiquement, le manque de formation est un frein significativement plus exprimé chez les praticiens estimant leur FI comme non suffisante et chez ceux n’ayant pas réalisé de FC (p > 0.01). - Manque d’intérêt Les médecins n’ayant pas réalisé de FC se sentent significativement moins concernés par la pratique de la gynécologie que leurs confrères qui se sont formés (p < 0.01). 41 89,5% des médecins ayant répondus avoir un manque d’intérêt pour cette discipline n’avaient pas effectué de FC. - Relation médecin – patientes / Gêne Il existe une différence significative entre le sentiment de gêne vis-à-vis de la patiente et le sexe du praticien (p < 0.01). 93,8% des médecins rencontrant ce frein étaient des hommes. - Manque de matériel / équipement Le profil du médecin n’exerce aucune influence significative sur réponses concernant ce frein. - Manque de rémunération Aucune différence significative n’a été mise en évidence selon les caractéristiques des médecins pour ce frein.	2,842
https://openalex.org/W4387501996	OpenAlex	Open Science	CC-By	2,023	Large mesenchymal cystic and chondroid pulmonary hamartoma mimicking lung cancer: Case report	Seha Ahn	English	Spoken	2,573	4,767	© The Author(s) 2023. 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 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. CASE REPORT Open Access Journal of Cardiothoracic Surgery Journal of Cardiothoracic Surgery Journal of Cardiothoracic Surgery Ahn et al. Journal of Cardiothoracic Surgery (2023) 18:278 https://doi.org/10.1186/s13019-023-02394-z Ahn et al. Journal of Cardiothoracic Surgery https://doi.org/10.1186/s13019-023-02394-z Abstract Pulmonary hamartoma is the most commonly resected benign neoplasm of lung. The mesenchymal cystic subtype is a rare and often bilaterally occurring variant composed of multiple cysts and nodules. Herein, we present an asymptomatic 70-year-old woman with a large and mostly cystic growth of right hilar region. Computed tomography of the chest and fluorodeoxyglucose positron emission tomography/computed tomography imaging traced its origins to right middle lobe. Overall features suggested primary lung cancer or perhaps other cystic lung disease. Because transbronchial lung biopsy failed to establish a histologic diagnosis, right middle lobectomy was undertaken by video-assisted thoracoscopic surgery. The gross surgical specimen harbored a single and sizeable (8.0 × 4.0 cm) cystic lesion containing multiple yellow-white nodules. A diagnosis of mesenchymal cystic and chondroid hamartoma was ultimately rendered. This particular case is noteworthy, given the initial clinical resemblance to primary lung cancer. Keywords Cystic hamartoma, Chondroid hamartoma, Mimicking, Lung cancer Introduction bilaterally. These tumors grow slowly and tend to become cystic at diameters > 1 cm [2].h Pulmonary hamartoma is the most commonly resected benign lung tumor, affecting 0.025–0.032% of the adult population [1]. Typically, such growths replicate nor mal lung constituents, including fat, mucinous fibrous connective tissue, ciliated epithelium, and respiratory epithelial-lined adenoid structures (2). Mesenchymal cys tic hamartoma (MCH) is a rare variant first reported in 1986. It is largely comprised of primitive mesenchymal cells that form multiple cysts and nodules, often arising The elderly woman we describe was initially thought to have primary lung cancer with cystic change. Instead, a mesenchymal cystic and chondroid hamartoma was con firmed through surgical resection. Large mesenchymal cystic and chondroid pulmonary hamartoma mimicking lung cancer: Case report Seha Ahn1, Heejin Lee1, Joon Kyu Kang1, In Sub Kim1, Youngkyu Moon1, Jung Suk Choi1 and Si Young Choi1* Case presentation An outpatient, a 70-year-old woman with no history of smoking, presented our Department of General Surgery with bilateral lower leg edema. Historically, a right lung nodule encountered 15 years prior had received no sub sequent follow-up. The patient has a medical history that includes hypertension, diabetes mellitus, and hyperlipid emia. In terms of past surgeries, she underwent total knee replacement surgery for both knees, with the right side operated on 5 months ago and the left side 11 years ago, *Correspondence: Si Young Choi redbaron@catholic.ac.kr 1Department of Thoracic & Cardiovascular Surgery. Eunpyeong St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, 1021, Tongil-ro, Eunpyeong-gu, Seoul 03312, Republic of Korea © The Author(s) 2023. 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 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. Page 2 of 5 Page 2 of 5 Ahn et al. Journal of Cardiothoracic Surgery (2023) 18:278 Ahn et al. Journal of Cardiothoracic Surgery additional 10-mm incisions of mid-axillary and posterior axillary lines at seventh ICS. Upon inspection, a thick- walled cavitary lesion of RML was noted. The resected specimen was collected by Endo Catch bag (Medtronic, Minneapolis, MN, USA) to retrieve via working port for frozen section diagnosis. Tissue assessed during medias tinal lymphadenectomy appeared benign, suggestive of hamartoma, so no further procedures were done. Opera tive time was 195 min, and the duration of anesthesia was 240 min. The estimated blood loss was 100 cc. both performed at a different institution. Chest X-ray now disclosed a cystic area and a ground-glass opac ity of the right hilar region (Fig. 1A). Computed tomog raphy (CT) of the chest localized both lesions (cystic: 8.8 × 6.1 cm; solid: 4.9 cm) to the right middle lobe (RML) (Fig. 1B, C), suggesting primary lung cancer with cystic change. She was referred to our Pulmonary Department for further evaluation. The patient was asymptomatic on physical exam. Her preoperative pulmonary function tests and echocardio gram results were within normal limits. In the absence of overt endobronchial pathology during bronchoscopy, a transbronchial lung biopsy and lavage fluid were rou tinely obtained. Fluorodeoxyglucose positron emission tomography/computed tomography (18 F-FDG PET/ CT) showed a large cystic area of RML, with mild FDG uptake. There was also spotty FDG uptake at right lower paratracheal and subcarinal locations and involving both hilar and peribronchial regions. The interpretation of 18 F-FDG PET/CT images in the right lower paratracheal and subcarinal regions indicated reactive lymph nodes, leading to a diminished suspicion of lymph node metas tasis. Despite favoring a chronic inflammatory process with cyst formation, efforts to exclude malignancy were warranted. The transbronchial lung biopsy was not help ful (respiratory epithelium only), and the inordinate risk of pneumothorax precluded CT-guided percutaneous cutting needle biopsy (PCNB). Diagnosis and treatment were then entrusted to our Department of Thoracic and Cardiovascular Surgery. h An upright anteroposterior chest X-ray obtained in the recovery room (20 min after procedural completion) was clear (Fig. 2). As depicted radiographically, the resected lung specimen (Fig. 3) grossly displayed a single large cystic mass (13.0 × 9.0 × 4.5 cm) containing multiple yel low-white nodules. A mesenchymal cystic and chondroid hamartoma was confirmed by the final pathology report. Additionally, the resection margin was clear, and the pleural surface exhibited a smooth and glistening appear ance. Furthermore, the paratracheal lymph node and hilar lymph node were negative, indicating the absence of cancer involvement. The chest tube was removed on postoperative Day 5, and no complications were observed in the patient. However, to ensure better pain control, the patient was discharged on postoperative Day 9. Her clini cal course thereafter has been uneventful for 36 months. Discussion In this particular instance, the initially suspected primary lung cancer (with cystic change) proved to be a rare vari ant of pulmonary hamartoma. Various subtypes are cat egorized by the major component: cartilage (chondroid hamartoma, most common), smooth muscle, or con nective tissue [1]. In CT studies, hamartomas typically present as single, well-defined nodules that are smooth- surfaced and round or lobulated. The radiographic Right middle lobectomy was eventually performed by video-assisted thoracoscopic surgery (VATS). The patient was placed in left lateral decubitus position, using a double-lumen endotracheal tube for single-lung ven tilation. An incision (4.0 cm) was made in anterior axil lary line at fifth intercostal space (ICS), followed by two Fig. 1 Preoperative imaging studies: (A) cystic area (black arrow) and ground-glass opacity of right hilar region detected by chext X-ray; and (B, C) com puted tomography of chest (axial and sagittal views) demonstrating large cystic area (8.8 × 6.1 cm) and solid growth (4.9 cm) of right middle lobe Fig. 1 Preoperative imaging studies: (A) cystic area (black arrow) and ground-glass opacity of right hilar region detected by chext X-ray; and (B, C) com puted tomography of chest (axial and sagittal views) demonstrating large cystic area (8.8 × 6.1 cm) and solid growth (4.9 cm) of right middle lobe Fig. 1 Preoperative imaging studies: (A) cystic area (black arrow) and ground-glass opacity of right hilar region detected by chext X-ray; and (B, C) com puted tomography of chest (axial and sagittal views) demonstrating large cystic area (8.8 × 6.1 cm) and solid growth (4.9 cm) of right middle lobe Ahn et al. Journal of Cardiothoracic Surgery (2023) 18:278 (2023) 18:278 Page 3 of 5 The metabolic activity observed on 18 F-FDG PET/CT for pulmonary hamartoma typically appears absent or low, similar to other benign lung lesion [8]. Several fac tors contribute to the diagnosis of pulmonary hamar toma using 18 F-FDG PET/CT, such as the presence of a single lesion, a low maximum standardized uptake value (SUVmax) of the lesion (falling below the internationally recognized cut-off value of 2.5), and the absence of any other coexisting malignant tumors [8–10]. At our insti tution, outpatient follow-up is performed for suspected cases of typical hamartoma. In cases where a large mass is present and distinguishing it from primary lung can cer is challenging, 18 F-FDG PET/CT scans are con ducted. Data Availability Th d t d l i The data underlying this article will be shared on reasonable request to the corresponding author. Ethics approval and consent to participate The study was approved by the institutional review board of Eunpyeong St. Mary’s Hospital at the Catholic University of Korea with Reference and approval number (PC23ZESI0047). Acknowledgements Acknowledgements Not applicable. Not applicable. Informed consent In this report, informed consent to participate was obtained from the patient and her family. Abbreviations MCH Mesenchymal cystic hamartoma CT Computed tomography RML Right middle lobe 18F-FDG PET/CT Fluorodeoxyglucose positron emission tomography/ computed tomography PCNB percutaneous cutting needle biopsy VATS Video-assisted thoracoscopic surgery ICS Intercostal space Author contributions SA and HL, supported by SC, performed the literature search and wrote the preliminary manuscript. SC performed the surgery. SA assisted in the surgery. All authors reviewed the manuscript draft and revised it critically on intellectual content. All authors approved the final version of the manuscript to be published. Conclusionh This 70-year-old woman was initially thought to have pri mary lung cancer with cystic change. Tissue examination later revealed a rare variant of pulmonary hamartoma, specifically a mesenchymal cystic and chondroid subtype. Discussion If deemed necessary, PCNB or surgical resection hallmark of chondroid hamartoma is a fatty density with popcorn-like calcifications [1, 3]. i MCH is a rare and often bilateral growth in which largely primitive mesenchymal cells proliferate, forming multiple cysts and nodules [2]. Hemoptysis, pneumo thorax, hemothorax, pleuritic chest pain, or dyspnea, may or may not accompany these tumors [4, 5]. Discov ery by X-ray in our asymptomatic patient was inciden tal. The differential diagnosis includes other benign and radiographically similar diseases (i.e., pleuropulmonary blastoma, cystic adenomatoid malformation, and lymph angiomyomatosis), with malignant transformation as a remote possibility [4, 6, 7]. Ahn et al. Journal of Cardiothoracic Surgery (2023) 18:278 (2023) 18:278 Page 4 of 5 Fig. 3 Gross appearance of surgical specimen, demonstrating single, large cystic growth (8.0 × 4.0 cm) with multiple yellow-white nod rance of surgical specimen, demonstrating single, large cystic growth (8.0 × 4.0 cm) with multiple yellow-white nodules Standardized uptake value SUV is performed for further evaluation. In our specific case, a large solid tumor with a cystic component was a rare variant of pulmonary hamartoma, and the size of the lesion did not correlate with typical hamartoma charac teristics. Furthermore, it was essential to differentiate the patient’s large solid tumor with a cystic component from primary lung cancer. Despite the presence of mild FDG uptake on the 18 F-FDG PET/CT scan, accurate dis crimination between these two conditions was of utmost importance. CT-guided PCNB was not a viable option in the wake of an unproductive transbronchial lung biopsy. Cystic lesions carry a high risk of pneumothorax and obfuscate needle localization. Furthermore, withdrawal of fluid was likely, making accurate diagnosis difficult [11]. We finally settled on a surgical approach for diagno sis and treatment. Following RML resection, the patient has remained recurrence-free for 36 months. Consent for publication Informed consent to participate was obtained from the patient and her family. References 8. Ye S, Meng S, Bian S, Zhao C, Yang J, Lei W. Diagnosis value of 18F-Fluoro-d- glucose positron emission tomography-computed tomography in pulmo nary hamartoma: a retrospective study and systematic review. BMC Med Imaging. 2023;23(1):28. 8. Ye S, Meng S, Bian S, Zhao C, Yang J, Lei W. Diagnosis value of 18F-Fluoro-d- glucose positron emission tomography-computed tomography in pulmo nary hamartoma: a retrospective study and systematic review. BMC Med Imaging. 2023;23(1):28. References 1. Radosavljevic V, Gardijan V, Brajkovic M, Andric Z. Lung hamartoma-diagnosis and treatment. Med Archives. 2012;66(4):281. 2. Mark EJ. Mesenchymal cystic hamartoma of the lung. N Engl J Med. 1986;315(20):1255–9. 3. Leiter Herrán F, Restrepo CS, Alvarez Gómez DI, Suby-Long T, Ocazionez D, Vargas D. Hamartomas from head to toe: an imaging overview. Br J Radiol. 2017;90(1071):20160607. 4. Zhu H, Huang S, Zhou X. Mesenchymal cystic hamartoma of the lung. Ann Thorac Surg. 2012;93(6):e145–7. 5. Glezos J, Toppin D, Cooney T. Mesenchymal cystic hamartoma presenting with spontaneous pneumothorax. Can Respir J. 2003;10(5):280–1. 6. Ribet M, Jaillard-Thery S, Nuttens MC. Pulmonary hamartoma and malig nancy. J Thorac Cardiovasc Surg. 1994;107(2):611–4. 7. Fasanya AA, Hattab Y, Patel A, Lega M. Mesenchymal cystic hamartoma of the lung Respiratory Med case Rep 2017;21:158 60 9. Ergonul AG, Akcam TI, Özdil A, Turhan K, Cakan A, Cagirici U. Diagnostic value of 18 F-FDG-PET/CT in benign lung diseases. Kardiochirurgia i Torakochirurgia Polska/Polish Journal of Thoracic and Cardiovascular Surgery. 2018;15(1):1–4. 9. Ergonul AG, Akcam TI, Özdil A, Turhan K, Cakan A, Cagirici U. Diagnostic value of 18 F-FDG-PET/CT in benign lung diseases. Kardiochirurgia i Torakochirurgia Polska/Polish Journal of Thoracic and Cardiovascular Surgery. 2018;15(1):1–4. 10. Bunyaviroch T, Coleman RE. PET evaluation of lung cancer. J Nucl Med. 2006;47(3):451–69. 11. Yuan L, Wang S, Wei J, Yang K, Mao Y. Mesenchymal cystic hamartoma of the lung: a case report. Med (Baltim). 2022;101(1):e28242. 11. Yuan L, Wang S, Wei J, Yang K, Mao Y. Mesenchymal cystic hamartoma of the lung: a case report. Med (Baltim). 2022;101(1):e28242. 11. Yuan L, Wang S, Wei J, Yang K, Mao Y. Mesenchymal cystic hamartoma of the lung: a case report. Med (Baltim). 2022;101(1):e28242. Conflict of interest Received: 30 March 2023 / Accepted: 30 September 2023 Page 5 of 5 Page 5 of 5 Ahn et al. Journal of Cardiothoracic Surgery (2023) 18:278 (2023) 18:278 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. 7. Fasanya AA, Hattab Y, Patel A, Lega M. Mesenchymal cystic hamartoma of the lung. Respiratory Med case Rep. 2017;21:158–60.	17,764
https://openalex.org/W4281562471	OpenAlex	Open Science	CC-By	2,022	Fragility Fractures of the Acetabulum: Current Concepts for Improving Patients’ Outcomes	Giuseppe Toro	English	Spoken	7,935	15,099	Abstract The incidence of fragility fractures of the acetabulum (FFA) is constantly increasing. Generally, these fractures are related to a fall on the greater trochanter involving the anterior column. The management of FFA is extremely difficult considering both patients’ comorbidities and poor bone quality. Both non-operative and several operative treatment protocols are avail- able, and the choice among them is still ambiguous. The proposed surgical techniques for FFA [namely open reduction and internal fixation (ORIF), percutaneous fixation and total hip arthroplasty (THA)] are associated with a high complication rate. The treatment with the higher early mortality is the ORIF + THA, while the one with the lowest is the non-operative. However, at longer follow-up, this difference dreadfully change is becoming the opposite. Frequently ORIF, percutaneous fixation, and non-operative treatment need a subsequent re-operation through a THA. This latter could be extremely difficult, because of poor bone quality, acetabular mal union/non-union, bone gaps and hardware retention. However, the outcomes of each of the proposed treatment are mostly poor and controverted; therefore, a comprehensive patient evaluation and an accurate fracture description are required to appropriately manage acetabular fracture in the elderly. Keywords Acetabular fracture · Elderly · Fragility fracture · Open reduction and internal fixation · Total hip arthroplasty · Percutaneous fixation · Osteoporosis · Hip fracture · Mortality · Plate Fragility Fractures of the Acetabulum: Current Concepts for Improving Patients’ Outcomes Giuseppe Toro1,2 · Adriano Braile1 · Annalisa De Cicco1 · Raffaele Pezzella3 · Francesco Ascione4 · Antonio Benedetto Cecere1 · Alfredo Schiavone Panni1 Received: 18 July 2021 / Accepted: 4 May 2022 / Published online: 26 May 2022 © The Author(s) 2022 Indian Journal of Orthopaedics (2022) 56:1139–1149 https://doi.org/10.1007/s43465-022-00653-0 REVIEW ARTICLE * Giuseppe Toro giusep.toro@gmail.com * Giuseppe Toro giusep.toro@gmail.com 1 Department of Medical and Surgical Specialties and Dentistry, University of Campania “Luigi Vanvitelli”, 80138 Naples, Italy 2 Department of Clinical Sciences and Translational Medicine, University of Rome Tor Vergata, 00133 Rome, Italy 3 Unit of Orthopedics and Traumatology, AORN San Giuseppe Moscati, 83100 Avellino, Italy 4 Department of Orthopaedic and Traumatology Surgery, Ospedale Buon Consiglio Fatebenefratelli, 80123 Naples, Italy * Giuseppe Toro giusep.toro@gmail.com Introduction population one of the most commonly affected (about 24% of all acetabular fractures) [6]. As a definition, FFA are due to a fall from a standing height, with a subsequent impact on the greater trochanter. The resulting anteromedial force com- monly leads to a fracture of the anterior column and/or the quadrilateral plate with a medialization of the femoral head and a supero-medial roof impaction [6]. The management of FAA is still a matter of debate. Particularly, indications for non-operative versus operative treatment, the reliability of surgical fixation in an osteoporotic bone and the safety of complex joint reconstructive procedures (i.e., revision arthroplasties) are some of the unmet needs. Moreover, regardless of the treatment choice, final outcomes are mostly poor both in terms of function and mortality (Table 1). The aim of the present study is to aid the orthopaedic surgeon in the treatment decision making for fragility acetabular frac- tures through a comprehensive literature review, focusing on the technical tips that may aid to improve patient’s outcomes. The constant increase in life expectancy led to a growing incidence of fragility fractures [1–4]. Recently, a constant increase of fragility fractures of the acetabulum (FFA) has been observed [5]. Particularly, a 2.4-fold increase in the incidence of acetabular fractures in patients over 60 years of age during the last 3 decades was observed, making this 4 Department of Orthopaedic and Traumatology Surgery, Ospedale Buon Consiglio Fatebenefratelli, 80123 Naples, Italy :(0123 1 23456789) 3 Indian Journal of Orthopaedics (2022) 56:1139–1149 1140 Table 1 Treatment-related mortality risk. Adapted from Daurka et al.9 ORIF open reduction and internal fixation, THA total hip arthroplasty Procedure Mortality rate (%) Mortality follow-up (months) Conservative 12 52 ORIF 15.3 42.2 ORIF + THA 13.15 33.3 Percutaneous Fixation 30.5 121.8 investigating on both the femoral neck and the acetabu- lum, and to adequately manage the patient an appropri- ate evaluation of pre-fracture patients’ walking ability is recommendable. The clinics of a patient with an acetabular fracture depend on the fracture displacement, varying from the absence of pain and normal range of motion (ROM) to intractable pain, lower limb discrepancy and restricted ROM. In case of a suspected acetabular fracture, a standard- ized radiograph protocol, based on an anteroposterior (AP) and Judet oblique views (both obturator and iliac), must be obtained [11]. These X-rays are generally able to identify and classify the fracture. The Management of FFA In the elderly, the appropriate management of an acetabular fracture should consider not only the fracture pattern and the available treatment options, but also the patient’s health status and pre-fracture mobility. Therefore, a comprehen- sive patient’s history must be collected. Different treatment options are available both non-operative and operative. How- ever, recently, a shift in paradigm from the conservative to the operative treatment could be observed, with the final purpose of improving patient’s early mobility and lowering bed rest complications. Introduction However, CT scan is useful to improve fracture diagnosis and classification. In fact, some characteristics of the fractures (i.e., articular incongruities, nondisplaced fractures, intra-articular fragments, femoral head subluxation or quadrilateral plate impaction) are eas- ily observed using a CT scan. Moreover, CT with 3D recon- structions can help to visualize complex fractures and to plan the surgical procedure, being able to enhance diagnos- tic and therapeutic accuracy [12]. However, considering the mechanism of fracture and the poor bone quality, fragility acetabular fractures might be nondisplaced and difficult to diagnose. Therefore, in case of high suspicious of FFA with inconclusive X-ray and CT scans, an MRI or a bone scan should be used to identify the fracture [11, 13, 14]. ORIF open reduction and internal fixation, THA total hip arthroplasty Fig. 1 a A 3D reconstruction of a two columns fracture with the involvement of the quadrilateral plate occurred in a 75-year-old male. b Coronal reconstruction of a CT scan. Please note in red the “gull sing” that represent the result of the supero-medial impaction of the femoral head Fig. 1 a A 3D reconstruction of a two columns fracture with the involvement of the quadrilateral plate occurred in a 75-year-old male. b Coronal reconstruction of a CT scan. Please note in red the “gull sing” that represent the result of the supero-medial impaction of the femoral head Patient Evaluation and Fracture Pattern FFA presents some differences from acetabular fractures observed in the young. In fact, in the elderly, most of ace- tabular fractures are related to a lateral compression force on the greater trochanter transmitted antero-medially to the anterior column, the anterior wall and the quadrilateral plate (Fig. 1) [6–8]. This characteristic mechanism of frac- ture explains the high incidence anterior column fractures both elementary and associated [4]. Furthermore, because of poor bone quality, the FFA is associated to an increased incidence of both femoral head injury and posterior hip dislocation related to a more severe posterior wall involve- ment (i.e. marginal impaction or comminution) [5, 9, 10]. These observations underline the troublesome need of both appropriately diagnose and treat fragility acetabular frac- tures. As a rule, a comprehensive evaluation of an elderly patient after a fall from a standing height is mandatory, Non‑operative Treatment The non-operative treatment of elderly patients with FFA might be associated to poor outcomes, in terms of both bed rest complications and joint function. Therefore, it should be indicated only in selected fractures patterns and patients [15–17]. Patients with severe comorbidities or with a severely impaired pre-injury mobility are those most eli- gible for conservative management. Moreover, also mini- mally displaced fractures (including anterior column and transverse ones) could be managed non-operatively, thanks to the intrinsic fracture stability [17–19]. According Lovric 1 3 3 Indian Journal of Orthopaedics (2022) 56:1139–1149 1141 et al., functional outcomes and bed rest were lower in con- servatively treated transverse fractures than anterior column/ wall ones [20]. The relevance of fracture pattern in patient’s outcomes was further underlined by Heeg et al. in a study on 57 conservatively treated FFA [21]. In fact, the authors observed that patients with acetabular roof impaction and a displacement over 2 mm reported the worst outcomes [21]. However, fracture displacement should not be considered a mandatory factor for surgical indication. In fact, fractures with a secondary congruence of the hip joint (defined as congruence of the femoral head with the acetabular roof on antero-posterior and Judet views without traction [22]) might be considered for non-operative treatment. The sec- ondary congruence could be commonly observed in both column fractures, where a congruency between the femoral head and the acetabulum might be observed despite the lack of a continuity of the articular surface with the hemipelvis [23]. high-energy trauma [30]. However, considering the comor- bidities and the singular fracture pattern observable in the elderly, whenever possible non-extensile approaches should be preferred over combined and extensile ones, because of these latter are associated with longer surgical times and higher complications rate [31]. In fractures involving the anterior wall, the anterior column, or the quadrilateral plate, an ilioinguinal approach is generally preferred. However, the anterior column and the quadrilateral plate fractures might be treated using the Stoppa approach. On the other hand, the Kocher–Langenbeck approach is recommended for posterior column or posterior wall fractures [31]. p Considering the poor bone quality, commonly observed in primary osteoporosis as well as in other metabolic bone disease [32], fixation stability is another issue of concern in FFA. Some precautions might be used to overcome this issue. Non‑operative Treatment The addition of plate fixation to lag screws improves fixation stability, with locking plates that were specifically developed for the osteoporotic bone [4, 33, 34]. Although the cement augmentation is widely used for improving fixa- tion stability in the osteoporotic bone throughout the body [35, 36], to the best of our knowledge, it has not been used for acetabular fractures. However, nevertheless the type of plate used (conventional or locked), a buttress plate is rec- ommended in case of wall fractures, whereas a neutralization plate for column ones. In posterior column fractures, the use of two buttress plates provides a better fracture stability [37], probably also because the possibility of using a higher screw density in the areas of high bone density. The involve- ment of quadrilateral plate is a relevant issue considering that its management might be extremely difficult [38, 39]. Figure 2 shows a clinical case representing a failed fixa- tion due to quadrilateral plate reduction loosing. In fact, the quadrilateral plate needs an optimal reduction of the medial fragment protrusion to assure viable outcomes (see Fig. 3). A biomechanical analysis conducted by Culemann et al., analysed different types of fixation of the quadrilateral plate (including conventional, locking and specifically designed plates) showing that conventional plates with three periar- ticular screws provides the best fixation stability [40]. Any- way, quadrilateral plate non-anatomical reduction could be considered acceptable in elderly patients when the femoral head remains centered within the acetabular roof [41]. Skeletal traction should be avoided, considering the high the high risk of pin-related complications (i.e. pin site infections or pin pull-out), the unviable reduction due to the rotational deforming forces that acts on the acetabular frac- tures, the need of prolonged bed immobilization and poor bone quality [18, 24, 25]. Therefore, also those patients non- operatively treated should be early mobilized, while partial weight-bearing initiated as soon as possible depending on patient’ tolerance. However, a constant evaluation of the fracture stability through several radiographs is suggested during the entire follow-up period to early identify any sec- ondary displacement that may require a change in the man- agement [5, 14, 17]. Osteosynthesis Open Reduction and Internal Fixation (ORIF) The FFA are associated frequently associated to bad outcomes because of some of the conditions associated to the aging, like osteoporosis and low level of activity, are considered risk factors for poor results [26]. However, although the clini- cal outcomes are generally worse than those reported in the younger population, ORIF represents the gold standard of treatment for most displaced FFA [18]. Anyway, as a rule, considering patient’s comorbidities, fracture comminution, and poor bone quality, also a non-anatomic reduction can ensure optimal results in terms of both fracture healing and early mobilization, that represent mandatory goals in the elderly [27–29]. Percutaneous Fixation Percutaneous fixation of FFA is a challenging technique that could offer some advantages in a subset of patients. In fact, quick recovery, minimal blood loss and a low risk of post-operative infection are achievable, using appropriate small stab incisions to insert modified clamps and pushers for fracture’s reduction and cannulated screw for fixation [42, 43]. Another theoreti- cal advantage of percutaneous fixation is the possibility to perform a subsequent arthroplasty with less technical Obviously, the surgical plan of an ORIF of an FFA should start from the surgical approach. The surgical approaches available for the treatment of fragility acetabular frac- tures are the same described for those occurring after a 1 1142 Indian Journal of Orthopaedics (2022) 56:1139–1149 Fig. 2 a Anteroposterior X-ray view of a fragility both column frac- ture occurred in an 84-year-old female. Please note the degree of fracture fragmentation and the involvement of the quadrilateral plate. b and c Anteroposterior and oblique postoperative X-rays. Because of the patient started to complain respiratory failure during the pro- cedure, a non-anatomic reduction of the quadrilateral plate was accepted. d Anteroposterior X-ray at 1 month of the surgery showing reduction loosening of the quadrilateral plate and femoral head medi- alization Fig. 2 a Anteroposterior X-ray view of a fragility both column frac- ture occurred in an 84-year-old female. Please note the degree of fracture fragmentation and the involvement of the quadrilateral plate. b and c Anteroposterior and oblique postoperative X-rays. Because of the patient started to complain respiratory failure during the pro- cedure, a non-anatomic reduction of the quadrilateral plate was accepted. d Anteroposterior X-ray at 1 month of the surgery showing reduction loosening of the quadrilateral plate and femoral head medi- alization Fig. Osteosynthesis 3 An anteroposterior X-ray in a 78-year-old lady. As opposite of the previous case, note the anatomical reduction of the quadrilateral plate that led to fracture healing without the further development of the osteoarthritis Fig. 4 Antero-posterior standard X-ray, showing the three corridors for the percutaneous fixation of the acetabular fractures. In Blue, the iliac-pubic corridor (in retrograde fashion) for the anterior column. In Yellow, the iliac-ischiatic corridor for the fixation for the posterior column. In Red, the dome corridor Fig. 3 An anteroposterior X-ray in a 78-year-old lady. As opposite of the previous case, note the anatomical reduction of the quadrilateral plate that led to fracture healing without the further development of the osteoarthritis Fig. 4 Antero-posterior standard X-ray, showing the three corridors for the percutaneous fixation of the acetabular fractures. In Blue, the iliac-pubic corridor (in retrograde fashion) for the anterior column. In Yellow, the iliac-ischiatic corridor for the fixation for the posterior column. In Red, the dome corridor problems thanks to a limited scar tissue and soft tissue damage. The insertion of the percutaneous screws that fixes the bone is made through 3 osseous “corridors”: the iliac-pubic, the iliac-ischial and the dome zone (see Fig. 4 for further details). Generally, in case of simple column fracture, the stabilization of one corridor is sufficient to fix the fracture, while two to three corridors must be stabi- lized to fix complex fractures [43]. However, some impor- tant issues limit the use of these techniques. In fact, they need a considerable expertise and a relevant confidence with the radiological anatomy of the pelvis, considering the lack of direct visibility of the fracture site. Moreover, an accurate fracture reduction might be difficult to obtain and the screws might be far from the correct and safe positions for the fracture stabilization [12]. Specific con- traindications to the percutaneous fixation include poste- rior wall fractures with hip instability and lack of surgeon expertise with percutaneous pelvic fixation [43]. i Table 2 summarizes the available treatment options and their drawbacks in FFA. Osteosynthesis 1 3 1143 Indian Journal of Orthopaedics (2022) 56:1139–1149 Table 2 Type of treatment, possible drawbacks, and their solutions ORIF Open reduction and internal fixation Type of treatment Drawbacks Possible solutions Non-operative Bed rest complications Early mobilization; partial weight-bearing as soon as possible Secondary fracture displacement Routine radiograph evaluation; operative treatment Secondary osteoarthritis Total hip arthroplasty ORIF Poor bone quality Neutralization plates OR Locking plates Quadrilateral plate displacement Reduction and plate fixation with 3 periarticular screws OR specific designed plate fixation for quadrilateral plate Surgical-related complications in high-risk patients Accept non-anatomical reduction; Prefer non-extensile surgical approaches Secondary osteoarthritis Total hip arthroplasty Percutaneous fixation Technical demanding Proper knowledge of the radiological anatomy of the pelvis Inappropriate screw positioning Accurate fracture reduction Secondary osteoarthritis Total hip arthroplasty Hip Replacement Acute THA Associated with Fracture Fixation The main indications for acute THA associated with fracture fixation are acetabular dome impaction > 40%, concomitant femo- ral head impaction/neck fracture, acetabular comminution, concomitant HOA and/or multiple associated fractures [44– 46]. The goal of fracture fixation in this type of treatment is represented by a rigid stabilization of the fracture without considering the quality of reduction to assure good primary stability to the THA [46]. Typically, this combined pro- cedure could be performed through a Kocher–Lagenbeck Hip osteoarthritis (HOA) is another critical issue that the orthopaedic surgeon has to face up in FFA. In fact, in the elderly, HOA could represent both a late complication and a concomitant disease of the acetabular fracture. Therefore, the use of total hip arthroplasty (THA) is an option that has to be considered, in both the acute setting and fracture sequelae. In Table 3, a summary of the possible pitfalls asso- ciated with THA in FFA. Table 3 THA after FFA, possible drawbacks, and their solutions THA Total hip replacement, FFA Fragility Fracture of the Acetabulum, ORIF Open reduction and internal fixation Type of THA Drawbacks Possible solutions Acute with ORIF Inadequate cup stability Column reconstruction + multi-hole revision shell Bone loss Allograft/autograft Poor bone quality Porous metal cups Surgical-related complications in high-risk patients Non-anatomical reduction; proper patient selection Poor long-term implant survivorship Proper patient and implant selection Delayed after non-operative treatment THA in non-union or malunion Accurate evaluation of the preoperative CT scan + appropriate surgical approach + revision cages; plate fixation; bone graft; porous buttress augmentation devices Bone loss Revision cages; bone graft; porous buttress aug- mentation devices Poor femoral bone quality Cemented stems Delayed after operative treatment Infection Rule out a possible unknown infection preopera- tively (i.e.: perform biochemical evaluations) Scar tissue and avascularity of the soft tissues Appropriate surgical approach; proper patient selection Bone loss Revision cages; bone graft; porous buttress aug- mentation devices Hardware retention Plan to remove hardware coming on the way of THA Table 3 THA after FFA, possible drawbacks, and their solutions Possible solutions THA Total hip replacement, FFA Fragility Fracture of the Acetabulum, ORIF Open reduction and internal fixation 1 3 Indian Journal of Orthopaedics (2022) 56:1139–1149 1144 approach, but a secondary anterior approach should be done if an anterior fixation is also required. still relevant, and represented by acetabulum malalignment, severe bone loss and fracture non-union. Hip Replacement Therefore, also in case of the occurrence of an HOA after an FFA, conserva- tively treated rTHA facilities are required. Particularly, cages and bone grafts might be extremely useful [52]. The choice of the surgical approach depends on the surgeon expertise and the type of sequelae. In fact, in case of severe anatomi- cal alterations or instability of the pelvic ring due to fracture non-union (a condition that required the correction of the pelvic ring alignment before performing THA), modified Kocher–Langenbeck or iliofemoral might be preferred [17, 18, 54, 55].i i The acetabular reaming should be made carefully, con- sidering the poor bone quality. The bone defects observable after fracture reduction should be filled with allograft or autograft (i.e., femoral head morselized graft). A multi-hole revision acetabular shell could be useful to achieve addi- tional fixation. The more recently introduced porous metal cups provides sufficient primary stability ensuring good clinical results at mid-term follow-up [47]. The femoral stem implantation is done standardly, using both uncemented and or cemented stems depending on patient’ age, bone quality and surgeon preference [5, 18, 46]. Generally, a bed to chair transfer can be started from the first day after the surgery, while a partial weight-bearing can be allowed from the sec- ond with a progressive weight-bearing [29]. The filling of acetabular bone gaps is one of the most relevant problems observed in these patients, especially in case of central protrusion of the femoral head with the consequent possible excessive medialisation of the implant. Recently, the use of porous tantalum buttress augmentation and tantalum acetabular component has been proposed to decrease the need of cementation and massive bone grafting while increasing the implant stability [56–58]. Another pos- sibility, is to use autologous graft from the patient’s femoral head [8, 59]. The use of impacted morselised allograft and a cemented cup, eventually with a reinforcement ring, could be another viable option to face up the problem of the bone loss [60, 61]. However, the indication for this kind of surgery depends also on patient’s general health status. Acute THA has been demonstrated to provide benefit of immediate postoperative weight-bearing, reducing the risk of any thrombotic events, decubitus ulcers and pulmonary complications [48]. How- ever, the procedure might be very challenging and time con- suming. Hip Replacement Moreover, the 10-year survivorship of this kind if implants is inferior compared to that observed in patients who underwent to THA for primary HOA or avascular necrosis (AVN) [48]. Therefore, a strict patient’s selection is required to assure better outcomes of acute THA in fragil- ity acetabular fractures. On the femoral side, the use of cemented stems should be preferred because of the predisposition of premature loosen- ing consequently to the long period of inactivity after hip fracture that might lead to a disuse osteoporosis [18]. Delayed Total Hip Arthroplasty A delayed THA might be extremely difficult. Therefore, a meticulous pre-operative planning is mandatory. Any bone defect should be analysed using the CT scan [14], and any potential joint infection should be ruled out especially in case of previous ORIF or percutaneous fixation, evaluating at least the serum bio- markers (i.e. erythrocyte sedimentation rate, C-reactive pro- tein, white cell blood count) [49, 50] . The use of revision THA (rTHA) facilities and approaches might be required to face up all the issues related to a delayed THA. 3 Patient’s Outcomes However, some concerns had been raised around their long-term safety, considering that their use had been associated with osteonecrosis of the jaw and atypical femoral fractures [72–74]. Therefore, an accurate follow- up of patients under anti-resorptive drugs is mandatory to timely correct the therapy, if required [66, 75]. Anabolic drugs, like teriparatide or romosozumab, are generally used in non-responsive patients to antiresorptive therapy (i.e.: occurrence of a new fragility fractures in patients under anti-resorptive drugs) [76–78]. Moreover, the availability of anabolic drugs paved the way to the implementation of sequential therapies strategies (i.e. antiresorptive therapy first followed by an anabolic drug; anabolic therapy first fol- lowed by an antiresorptive drug, co-administration of both antiresorptive and anabolic agents) [78, 79]. Among them, the administration of an anabolic drug followed by an antire- sorptive one seems to be associated with the best outcomes in terms of fragility fracture prevention [78]. THA Total hip arthroplasty, HOA Hip Osteoarthritis authors regardless of the fixation technique used (22% ORIF vs 25% percutaneous). Despite performing THA in patients with acetabular fractures is a complex procedure, it provides significant improvement in pain and function. In fact, Stibolt et al. [52] in a review including 448 patients with a median age of 51.5, observed that delayed THA was associated to an increase of the mean Harris Hip Score from 41.5 to 87.6 at a mean of 82 months of follow-up. Interestingly, the use of anti-osteoporotic drugs (both vitamin D supplementation and bisphosphonates) in patients with FFA was associated to a reduction in the time to frac- ture healing [68]. According to the authors, the osteoporo- sis management as a part of a standardized protocol for the management of FFA focused also on the optimisation of medical comorbidities, early ambulation, and early hospital discharge to an appropriate facility, was associated to an improvement in functional outcomes [68]. However, the complication rate is generally high (i.e. infections, sciatic nerve palsy, dislocation, heterotopic ossi- fication) and THA in acetabular fracture was associated to an inferior 10-year survival compared to those with primary OA or AVN [64]. Finally, a recent study conducted by Glogovac et al. observed that performing the surgery within the first 48 h after the injury were not associated with mortality rates benefits in elderlies with acetabular fractures, in contrast to that reported for proximal femur fractures [65]. Patient’s Outcomes Confusing outcomes had been reported in FFA, further underlining the difficulties that the orthopaedic surgeon face up in these patients. However, some risk factors for both reoperation need and poor outcomes had been identified (see Table 4) [62, 63]. A systematic review performed by Daurka et al. [9], com- pared ORIF vs percutaneous, ORIF + THA vs ORIF/Percu- taneous and ORIF Vs Conservative. The authors observed several controversies in the achievable outcomes and com- plications [9]. Particularly, ORIF + THA was the treatment associated with the highest and earliest mortality rate, whereas percutaneous fixation presented a higher mortality rate than ORIF. This surprising observation might be related to the older age of the patients included in that group. These results were further confirmed by the direct comparison through the pooled data between ORIF with ORIF + THA (death occurred at 47.2 months vs 33.3 months respectively) and ORIF vs percutaneous (death occurred at 47.2 months vs 121.8 months respectively) (see Table 1 for further details). Of note, a high conversion rate to THA was reported by the rTHA and THA after ORIF share several issues, includ- ing longer operative time, higher blood loss and transfusion rate, presence of scar tissues, avascularity of soft tissues and bone, heterotopic ossification, retained hardware, acetabular deformity and bone loss, and high risk of infection [51]. Considering the above risk factors, the clinical outcomes are less favourable, and a higher revision rate has to be expected following a delayed THA compared to hip replacement for primary HOA [52, 53]. The occurrence of post-traumatic HOA or AVN with the development of a painful hip joint with a decreased quality of life is one of the drawbacks of non-operative treatment of acetabular fractures [52, 53]. Although in this scenario, there are no soft tissue alteration, the technical problems are 3 3 Indian Journal of Orthopaedics (2022) 56:1139–1149 1145 Table 4 Factors associated with worse outcomes THA Total hip arthroplasty, HOA Hip Osteoarthritis Patient-related Age Osteoporosis Low activity level Contralateral THA Concomitant HOA Fracture-related Roof impaction (gull sign) Quadrilateral plate comminution Posterior wall fragmentation Concomitant femoral head fracture Surgery-related Non-anatomical reduction (espe- cially of the quadrilateral plate) Time consuming procedure and denosumab are well established and safe antiresorptive drugs with proved efficacy in preventing fragility fractures [66, 67, 69–71]. Conclusion Acetabular fractures in the elderly are constantly increas- ing during last years and requires a comprehensive patient’s evaluation and advanced surgical skills. The treatment choice is exceptionally difficult, considering the contro- verted outcomes and high rate of complications reported in the available literature. ORIF could provide good clinical outcomes when a congruent, anatomic reduction is obtained. However, an intermediate mortality risk had been associ- ated with ORIF and a large part of patients would require a subsequent THA. Patient’s Outcomes The authors concluded that the time to surgery in acetabular fragility fractures should be determined on individual bases [65]. In Fig. 5, we proposed an algorithm for a practical approach to FFA. Declarations Conflict of Interest Authors declare that they had no conflict of inter- est. Informed Consent Additional informed consent was obtained from all patients for whom identifying information is included in this article. Ethical Standard statement This article does not contain any studies with human or animal subjects. Open Access This article is licensed under a Creative Commons Attri- bution 4.0 International License, which permits use, sharing, adapta- tion, 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/. FFA and Osteoporosis Management Very few studies evaluated the efficacy of anti-osteoporotic drugs in FFA, however, as well as the other fragility frac- tures an appropriate management must also consider the underlying osteoporosis and sarcopenia to improve outcomes and prevent subsequent fractures [66–68]. According to the clinical guidelines of the Italian Society for Orthopaedics and Traumatology to appropriately manage a patient with an osteoporotic fracture, a comprehensive evaluation of the clinical risk factors, and an evaluation of the bone metabo- lism through both biochemical testing and DXA scan might be performed [66]. After excluding secondary osteoporo- sis, the treatment would be based mainly on vitamin D and/ or calcium supplementations and both antiresorptive and anabolic anti-osteoporotic drugs [66]. Bisphosphonates A primary THA with a concomitant ORIF might be pref- erable for patients with simple fractures and severe comor- bidities for whom a second surgery could be not affordable. However, this kind of approach was associated to the earliest mortality. Although theoretically less risky for the patient, a spe- cial attention in the preoperative planning is mandatory in case of delayed THA, because of the high complexity of 1 1146 Indian Journal of Orthopaedics (2022) 56:1139–1149 on a standardized protocol to ensure a holistic approach to FFA, with the final purpose of improve patients’ outcomes, may be advisable. Author Contributions AB, GT, and RP conceived the study. RP, ADC and ABC performed literature research. GT, FA, and ASP analyzed retrieved data. AB, ADC and GT wrote the article. GT and ASP super- vised the entire study. All authors read and approved the final draft of the article. Funding Open access funding provided by Università degli Studi della Campania Luigi Vanvitelli within the CRUI-CARE Agreement. None. 5. Butterwick, D., Papp, S., Gofton, W., Liew, A., & Beaule, P. E. (2015). Acetabular fractures in the elderly. JBJS, 97(9), 11. 3 References Fractures of the acetabulum: Accuracy of reduction and clinical results in patients managed operatively within three weeks after the injury. The Journal of Bone and Joint Surgery American, 78(11), 1632–1645. 8. Jouffroy, P. (2014). Indications and technical challenges of total hip arthroplasty in the elderly after acetabular fracture. Orthopae- dics & Traumatology: Surgery & Research, 100(2), 197–201. 27. Helfet, D. L., Borrelli, J., DiPasquale, T., & Sanders, R. (1992). Stabilization of acetabular fractures in elderly patients. The Jour- nal of Bone and Joint Surgery American, 74(5), 753–765.f 9. Daurka, J. 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https://openalex.org/W4225964674	OpenAlex	Open Science	CC-By	2,022	Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET	Dharmender Nishad	English	Spoken	16,218	34,753	Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 5 6 7 8 Dharmender · Kaushal Nigam · Satyendra Kumar 0 1 TFET demonstrates lesser sensitivity towards tempera- ture variation. Hence, the proposed DMSGO-SDP-TFET can be a suitable candidate for low power switching, and biosensing applications at elevated temperatures as compared to conventional DMSGO-TFET. Keywords Dielectric pocket · Temperature sensitiv- ity · stack gate-oxide · Linearity Research Article Keywords: Dielectric pocket, Temperature sensitivity, stack gate-oxide, Linearity Posted Date: January 12th, 2022 DOI: https://doi.org/10.21203/rs.3.rs-916159/v2 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Journal of Computational Electronics manuscript No. (will be inserted by the editor) Journal of Computational Electronics manuscript (will be inserted by the editor) Temperature Sensitivity Anal Gate Oxide Source Dielectric Dharmender · Kaushal Nigam · Satyendra Kuma Received: date / Accepted: date Abstract Temperature dependence performance vari- ation is one of the major concerns in predicting the ac- tual electrical characteristics of the device as the bandgap of semiconducting material varies with temperature. Therefore, in this article, for the ﬁrst time the impact of temperature variations ranging from 300K to 450K on the DC, analog/ radio frequency, and linearity per- formance of dual material stack gate oxide-source di- electric pocket-tunnel-ﬁeld-eﬀect transistor (DMSGO- SDP-TFET) is investigated. In this regard, technology computer-aided design (TCAD) simulator is used to analyze DC, and analog/radio frequency performance parameters such as carrier concentration, energy band variation, band-to-band tunneling rate (BTBT), IDS − VGS characteristics, transconductance (gm), cut oﬀfre- quency (fT ), gain-bandwidth product (GBP), maxi- mum oscillating frequency (fmax), transconductance fre- quency product (TFP), and transit time (τ) consider- ing the impact of temperature variations. Furthermore, linearity parameters such as third-order transconduc- tance (gm3), third-order voltage intercept point (VIP3), third-order input-interception point (IIP3), and inter- modulation distortion (IMD3) are also analyzed with temperature variations as these performance param- eters are signiﬁcant for linear and analog/radio fre- quency applications. Moreover, the performance of the proposed DMSGO-SDP-TFET is compared with the conventional dual-material stack gate oxide-tunnel-ﬁeld- eﬀect transistor (DMSGO-TFET). From the compara- tive analysis, in terms of % per kelvin, DMSGO-SDP- Dharmender (corresponding author) Department of Electronics and Communication Engineering, Jaypee Institute of Information Technology, Noida, India E-mail: dharmender.nishad@gmail.com 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 1 Introduction To overcome the scaling issues of conventional MOS- FETs, a tunnel ﬁeld-eﬀect transistor (TFET) based on the quantum tunneling mechanism has emerged as an alternative device to conventional MOSFETs due to lower subthreshold swing (SS) below 60 mV/decade, immunity to various short channel eﬀects, and low OFF- state current (IOF F ) [1]-[5]. However, TFETs have been reported some major limitations such as ambipolar cur- rent, lower ON-state current (ION) and poor analog/radio frequency performance due to ineﬃcient band-to-band tunneling [6]. Therefore, to overcome the lower ON- state current issue, various methods have been reported by the researchers, such as double-gate TFET, work- function engineering, hetero-dielectric, stacked gate struc- ture, electrically doped (ED), pocket doping, dielectric pocket, Extended Source TFET, dual material and gate over source overlap [6]-[22]. Furthermore, to address the ambipolar current issue, various methods have been reported such as, hetero-dielectric, work-function en- gineering, stacked gate structure, pocket doping, dual material gate [23]-[25]. In addition to the above issues, performance variation with temperature is also one of the major causes of concern in TFETs. Several studies [26]-[30] have reported temperature dependence perfor- mance of various TFET structures. 65 2 Dharmender et al. In the previous work [21], to enhance the DC, ana- log/radio frequency performance of the device, authors have proposed DMSGO-SDP-TFET with optimized di- electric pocket at the source-channel junction and re- ported an improved performance in terms of higher ON- state current (ION = 1.47 ×10−4 A) at (VGS = VDS =1.0 V), smaller point subthreshold swing (SS) = 15.7 mV/decade, maximum ION/IOF F ratio (3.14×1012), transconductance (gm = 1.02 ×10−3 S), and cut-oﬀfre- quency (fT = 193 GHz). This motivated us to further investigate the reliability of the proposed device at dif- ferent temperatures. For this, in this work, we inves- tigate the temperature sensitivity of DMSGO-TFET and DMSGO-SDP-TFET in terms of DC character- istics, analog/radio frequency, and linearity distortion performance parameters using TCAD simulations. leakage current, improve the switching ratio, subthresh- old swing and analog /radio frequency performance of the device. The temperature sensitivity of the proposed DMSGO- SDP-TFET is analyzed using TCAD simulations. For this, suitable models are incorporated such as the non- local band-to-band tunneling (BTBT) model is consid- ered to measure the tunneling probability across the junctions. The Shockley-Read-Hall model is enabled to account for the minority carrier recombination eﬀect. 1 Introduction The bandgap narrowing model is used to consider the bandgap narrowing caused by high doping concentra- tion. In our simulations, the Quantum conﬁnement ef- fect is not taken into account as it is signiﬁcant only if the thickness of the Si body is less than 10 nm. In addition to these models, Newton’s method was used to provide strong coupling between the resulting equations in order to improve current convergence. The remaining part of this paper is organized as follows. Section 2 presents the structural and simula- tion setup details. Section 3 describes the temperature sensitivity of DMSGO-TFET and DMSGO-SDP-TFET in three parts. The ﬁrst part presents a temperature sensitivity analysis of various DC parameters, the sec- ond part investigates the analog/radio frequency per- formance and the third part presents the linearity and distortion performance at diﬀerent temperatures. Fi- nally, the main ﬁndings of this work are concluded in Section 4. 19 20 21 22 23 24 25 26 27 28 29 30 Fig. 1 2-D schematic view of (a) conventional DMSGO- TFET and (b) DMSGO-SDP-TFET [21] 2 Device structure, parameters and simulation details The 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 Table 1 List of device parameters Parameters DMSGO-TFET DMSGO-SDP-TFET Gate Length (LG) 50 nm 50 nm SiO2 thickness (TSiO2) 0.8 nm 0.8 nm HfO2 thickness (THfO2) 1.2 nm 1.2 nm Pocket thickness - 2 nm Pocket height - 4 nm Silicon Film Thickness (Tsi) 10 nm 10 nm Channel doping (Nch) 1×1017 cm−3 1×1017 cm−3 Source doping(p type) (NS) 1×1020 cm−3 1×1020 cm−3 Drain doping(n type) (ND) 5×1018 cm−3 5×1018 cm−3 HfO2 dielectric constant (K) 25 25 Tunnel gate Length (L1) 10 nm 10 nm Control gate Length (L2) 25 nm 25 nm Auxiliary gate Length (L3) 15 nm 15 nm Tunnel gate workfunction(φ1) 4.0 eV 4.0 eV Control gate workfunction(φ2) 4.4 eV 4.4 eV Auxiliary gate workfunction(φ3) 4.0 eV 4.0 eV 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 energy bandgap of the semiconductor material decreases as temperature increases, inﬂuencing device character- istics accordingly. The energy bandgap variation with temperature can be obtained using the equation [30],[32] 3.1 Temperature sensitivity analysis of DC Parameters 2 3 4 e Eg(T) = Eg(0) −αT 2 T + β (2) This section presents the comparative temperature-sensit performance analysis of conventional DMSGO-TFET and proposed DMSGO-SDP-TFET in terms of carrier concentration, energy band variation, BTBT Variation, IDS −VGS characteristics, threshold voltage variation (VT ) and average subthreshold swing variation at ele- vated temperatures ranging from 300K to 450K. 25 26 27 28 29 30 31 32 (2) Where Eg(T) represents energy band gap at abso- lute temperature T, Eg(0) is the energy bandgap at T= 0 K, and α, β represents material-speciﬁc ﬁtting parameters. The ON-state energy band variation with temperature ranging from 300K to 450K for DMSGO- TFET and DMSGO-SDP-TFET are illustrated in Figs. 2(c) and 2(d), respectively. These results demonstrate that at room temperature, a signiﬁcant decrease in tun- neling barrier width (≈4nm) is observed at the source junction for DMSGO-SDP-TFET, which increases the inter-band tunneling rate for DMSGO-SDP-TFET as compared to conventional DMSGO-TFET. 2 Device structure, parameters and simulation details 2(c) and 2(d), respectively. The energy bandgap of the semiconductor material d as temperature increases, inﬂuencing device ch istics accordingly. The energy bandgap variat temperature can be obtained using the equation Eg(T) = Eg(0) −αT 2 T + β Where Eg(T) represents energy band gap lute temperature T, Eg(0) is the energy ban T= 0 K, and α, β represents material-speciﬁ parameters. The ON-state energy band variat temperature ranging from 300K to 450K for D TFET and DMSGO-SDP-TFET are illustrated 2(c) and 2(d), respectively. These results dem that at room temperature, a signiﬁcant decreas neling barrier width (≈4nm) is observed at th junction for DMSGO-SDP-TFET, which incre inter-band tunneling rate for DMSGO-SDP-T compared to conventional DMSGO-TFET. Furt the impact of temperature variation is noted small in both devices. Figs. 2(e) and 2(f) illustrate the inter-band ing rate with temperature ranging from 300K for DMSGO-TFET and DMSGO-SDP-TFET tively. The above ﬁgures show that, with tem variation, the inter-band tunneling rate increase the devices as shown in the inset. Furthermore demonstrate that the inclusion of a low k d pocket exhibits a higher tunneling rate due crease in tunneling width at the source-chann tion for DMSGO-SDP-TFET compared to conv DMSGO-TFET. Figs. 3(a) and 3(b) illustrate the eﬀect of t ture variations on IDS −VGS characteristics for tional DMSGO-TFET and DMSGO-SDP-TFE tively. 2 Device structure, parameters and simulation details These ﬁgures show a smaller ION variat 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 Table 1 List of device parameters Parameters DM Gate Length (LG) SiO2 thickness (TSiO2) HfO2 thickness (THfO2) Pocket thickness Pocket height Silicon Film Thickness (Tsi) Channel doping (Nch) 1× Source doping(p type) (NS) 1× Drain doping(n type) (ND) 5× HfO2 dielectric constant (K) Tunnel gate Length (L1) Control gate Length (L2) Auxiliary gate Length (L3) Tunnel gate workfunction(φ1) Control gate workfunction(φ2) Auxiliary gate workfunction(φ3) 3 Results and discussion 3.1 Temperature sensitivity analysis of DC Parameters This section presents the comparative temperature-sensitiv performance analysis of conventional DMSGO-TFET and proposed DMSGO-SDP-TFET in terms of carrier concentration, energy band variation, BTBT Variation, IDS −VGS characteristics, threshold voltage variation (VT ) and average subthreshold swing variation at ele- vated temperatures ranging from 300K to 450K. Figs. 2(a) and 2(b) depict the ON-state carrier con- centration variation with temperature ranging from 300K to 450K for conventional DMSGO-TFET and DMSGO- SDP-TFET, respectively. It is evident from these ﬁg- ures, due to stack gate oxide with lower work functions (φ1 = φ3 = 4.0 eV), at the source (drain) junctions, the electron (hole) concentration increases (decreases) in the channel, respectively for both the devices. More- over, as the temperature rises above room tempera- ture, the covalent bond inside the lattice of semicon- ductor body material begins to break, resulting in a larger number of electron-hole pairs (EHPs) generation. The rate of EHPs generation is directly propositional to the intrinsic carrier concentration of semiconductor (ni) [29]. The ni is exponentially related to temperature as per the expression ni = Naexp −Eg 2KT (1) Here, Na represents the impurity (acceptor/donor) car- rier concentration, Eg represents bandgap, K represents the Boltzmann constant, and T is the temperature . To understand the BTBT process the energy band variation of DMSGO-TFET and DMSGO-SDP-TFET are illustrated in Figs. 2(c) and 2(d), respectively. 2 Device structure, parameters and simulation details Figs. 1(a) and 1(b) illustrate the 2D structural views of conventional DMSGO-TFET and the proposed DMSGO- SDP-TFET for the parameters listed in Table 1. A stack gate oxide approach (low-k/high-k stack gate oxide i.e. SiO2/HfO2 stack) is used to provide better quality oxide/channel interface which enhances the ON-state current [30]. The entire length of the stack gate (LG) is considered 50 nm with SiO2 oxide layer thickness of (0.8 nm) and HfO2 oxide layer thickness of (1.2 nm) [19]. Furthermore, the total gate length (LG) of both devices is partitioned into three parts: tunneling gate (M1), control gate (M2), and auxiliary gate (M3) with diﬀerent length (L1, L2, L3) and work function (φ1, φ2, φ3), respectively. The control gate workfunc- tion (φ2) is considered 4.4 eV which corresponds to the metal molybdenum (Mo) (4.36 eV - 4.95 eV). The work functions (φ1 and φ3) are considered 4.0 eV which cor- responds to metal aluminum (Al) (4.0 eV - 4.26 eV) [24]. The combination of stack gate oxide approach with workfunction engineering (φ1 = φ3 = 4.0 eV < φ2 = 4.4 eV) and dielectric pocket at the source- channel in- terface is used to enhance the ON-state current, reduce 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 Fig. 2 Device structure, parameters and simulation details 1 2-D schematic view of (a) conventional DMSGO- TFET and (b) DMSGO-SDP-TFET [21] 65 3 Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 3 Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET Table 1 List of device parameters Parameters DMSGO-TFET DMSGO-SDP-TFET Gate Length (LG) 50 nm 50 nm SiO2 thickness (TSiO2) 0.8 nm 0.8 nm HfO2 thickness (THfO2) 1.2 nm 1.2 nm Pocket thickness - 2 nm Pocket height - 4 nm Silicon Film Thickness (Tsi) 10 nm 10 nm Channel doping (Nch) 1×1017 cm−3 1×1017 cm−3 Source doping(p type) (NS) 1×1020 cm−3 1×1020 cm−3 Drain doping(n type) (ND) 5×1018 cm−3 5×1018 cm−3 HfO2 dielectric constant (K) 25 25 Tunnel gate Length (L1) 10 nm 10 nm Control gate Length (L2) 25 nm 25 nm Auxiliary gate Length (L3) 15 nm 15 nm Tunnel gate workfunction(φ1) 4.0 eV 4.0 eV Control gate workfunction(φ2) 4.4 eV 4.4 eV Auxiliary gate workfunction(φ3) 4.0 eV 4.0 eV 3 Results and discussion 3.1 Temperature sensitivity analysis of DC Parameters This section presents the comparative temperature-sensitive performance analysis of conventional DMSGO-TFET and proposed DMSGO-SDP-TFET in terms of carrier concentration, energy band variation, BTBT Variation, IDS −VGS characteristics, threshold voltage variation (VT ) and average subthreshold swing variation at ele- vated temperatures ranging from 300K to 450K. Figs. 2(a) and 2(b) depict the ON-state carrier con- centration variation with temperature ranging from 300K to 450K for conventional DMSGO-TFET and DMSGO- SDP-TFET, respectively. It is evident from these ﬁg- ures, due to stack gate oxide with lower work functions (φ1 = φ3 = 4.0 eV), at the source (drain) junctions, the electron (hole) concentration increases (decreases) in the channel, respectively for both the devices. More- over, as the temperature rises above room tempera- ture, the covalent bond inside the lattice of semicon- ductor body material begins to break, resulting in a larger number of electron-hole pairs (EHPs) generation. The rate of EHPs generation is directly propositional to the intrinsic carrier concentration of semiconductor (ni) [29]. The ni is exponentially related to temperature as per the expression ni = Naexp −Eg 2KT (1) Here, Na represents the impurity (acceptor/donor) car- rier concentration, Eg represents bandgap, K represents the Boltzmann constant, and T is the temperature . To understand the BTBT process the energy band variation of DMSGO-TFET and DMSGO-SDP-TFET are illustrated in Figs. 2 Device structure, parameters and simulation details Furthermore, the impact of temperature variation is noted as very small in both devices. Figs. 2(e) and 2(f) illustrate the inter-band tunnel- ing rate with temperature ranging from 300K to 450K for DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. The above ﬁgures show that, with temperature variation, the inter-band tunneling rate increases in both the devices as shown in the inset. Furthermore, results demonstrate that the inclusion of a low k dielectric pocket exhibits a higher tunneling rate due to a de- crease in tunneling width at the source-channel junc- tion for DMSGO-SDP-TFET compared to conventional DMSGO-TFET. (1) Figs. 3(a) and 3(b) illustrate the eﬀect of tempera- ture variations on IDS −VGS characteristics for conven- tional DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. These ﬁgures show a smaller ION variation with Figs. 3(a) and 3(b) illustrate the eﬀect of tempera- ture variations on IDS −VGS characteristics for conven- tional DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. These ﬁgures show a smaller ION variation with To understand the BTBT process the energy band variation of DMSGO-TFET and DMSGO-SDP-TFET are illustrated in Figs. 2(c) and 2(d), respectively. The 65 Dharmender et al. 4 4 Dharmender et al. 2 Device structure, parameters and simulation details 0.00 0.02 0.04 0.06 0.08 0.10 -5 0 5 10 15 20 25 (a) M ajority carrier con cen tration D M S G O -TF ET V G S =1 .2 V , V D S =1 .0 V C ar r ie r c onc e ntr ation x 10 X (c m -3 ) P osition along c hanne l (µm) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K O N sta te electron h ole M in ority carrier con cen tration 0.00 0.02 0.04 0.06 0.08 0.10 -5 0 5 10 15 20 25 (b) M ajority carrier con cen tration M in ority carrier con cen tration D M S G O -S D P -T F E T V G S =1 .2 V ,V D S =1 .0 V C ar r ie r c onc e ntr ation x 10 X (c m -3 ) P osition along c hanne l (µm) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K O N sta te electron h ole 0.00 0.02 0.04 0.06 0.08 0.10 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 (c ) D M S G O -T F E T V GS = 1.2 V , V D S = 1.0V SO U R C E CH ANNE L DRAIN E n erg y (eV ) P o sitio n a lo n g ch a n n el (µm) 300K 350K 400K 450K O N sta te V B CB T unneling ba rrier w idth (~ 5 nm) e - 0.00 0.02 0.04 0.06 0.08 0.10 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 (d) e - T unneling ba rrier w idth (~ 4 nm) D M S G O -S D P -TF ET V GS = 1.2 V , V D S = 1.0V SO U R C E C H A N N E L D R A IN E ne r gy (e V ) P osition along c hanne l (µm) 300K 350K 400K 450K O N sta te V B CB 0 .0 0 0 .0 2 0 .0 4 0 .0 6 0 .0 8 0 .1 0 0 5 10 15 20 25 30 35 0 .0 6 5 9 0 0 .0 6 5 9 1 0 .0 6 5 9 2 0 .0 6 5 9 3 0 .0 6 5 9 4 3 0 .2 3 0 .3 3 0 .4 3 0 .5 D M S G O -TF ET Position alon g ch an n el (µm) Ban d to Ban d Tu n n elin g R ate (/cm 3 S) 300K 350K 400K 450K (e) V D S = 1.0V P o s itio n a lo ng cha nnel (µm) B a nd to B a nd T unne ling Ra te (/c m 3 S) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0 .0 0 0 .0 2 0 .0 4 0 .0 6 0 .0 8 0 .1 0 0 5 10 15 20 25 30 35 0 .0 6 5 9 1 0 .0 6 5 9 2 0 .0 6 5 9 3 3 0 .4 3 0 .5 3 0 .6 3 0 .7 D M S G O -S D P -TF ET Position alon g ch an n el (µm) Ban d to Ban d Tu n n elin g R ate (/cm 3 S) 300K 350K 400K 450K V D S = 1.0V (f) P o s itio n a lo ng cha nnel (µm) B a n d to B a n d T u n n elin g R a te (/cm 3 S) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K Fig. 2 Device structure, parameters and simulation details 2 Variation of carrier concentration with temperature for (a) DMSGO-TFET and (b) DMSGO-SDP-TFET, Variation of energy band with temperature for (c) DMSGO-TFET and (d) DMSGO-SDP-TFET, BTBT Variation with temperature for (e) DMSGO-TFET and (f) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 F en (e te o te O d cr d 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 1 0.00 0.02 0.04 0.06 0.08 0.10 -5 0 5 10 15 20 25 (a) M ajority carrier con cen tration D M S G O -TF ET V G S =1 .2 V , V D S =1 .0 V C ar r ie r c onc e ntr ation x 10 X (c m -3 ) P osition along c hanne l (µm) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K O N sta te electron h ole M in ority carrier con cen tration 0.00 0.02 0.04 0.06 0.08 0.10 -5 0 5 10 15 20 25 (b) M ajority carrier con cen tration M in ority carrier con cen tration D M S G O -S D P -T F E T V G S =1 .2 V ,V D S =1 .0 V C ar r ie r c onc e ntr ation x 10 X (c m -3 ) P osition along c hanne l (µm) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K O N sta te electron h ole 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 (a) g (b) g 0.00 0.02 0.04 0.06 0.08 0.10 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 (c ) D M S G O -T F E T V GS = 1.2 V , V D S = 1.0V SO U R C E CH ANNE L DRAIN E n erg y (eV ) P o sitio n a lo n g ch a n n el (µm) 300K 350K 400K 450K O N sta te V B CB T unneling ba rrier w idth (~ 5 nm) e - 0.00 0.02 0.04 0.06 0.08 0.10 -2.5 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 (d) e - T unneling ba rrier w idth (~ 4 nm) D M S G O -S D P -TF ET V GS = 1.2 V , V D S = 1.0V SO U R C E C H A N N E L D R A IN E ne r gy (e V ) P osition along c hanne l (µm) 300K 350K 400K 450K O N sta te V B CB - - - E n erg y (eV ) (d) g (c ) g 0 .0 0 0 .0 2 0 .0 4 0 .0 6 0 .0 8 0 .1 0 0 5 10 15 20 25 30 35 0 .0 6 5 9 1 0 .0 6 5 9 2 0 .0 6 5 9 3 3 0 .4 3 0 .5 3 0 .6 3 0 .7 D M S G O -S D P -TF ET Position alon g ch an n el (µm) Ban d to Ban d Tu n n elin g R ate (/cm 3 S) 300K 350K 400K 450K V D S = 1.0V (f) P o s itio n a lo ng cha nnel (µm) B a n d to B a n d T u n n elin g R a te (/cm 3 S) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0 .0 0 0 .0 2 0 .0 4 0 .0 6 0 .0 8 0 .1 0 0 5 10 15 20 25 30 35 0 .0 6 5 9 0 0 .0 6 5 9 1 0 .0 6 5 9 2 0 .0 6 5 9 3 0 .0 6 5 9 4 3 0 .2 3 0 .3 3 0 .4 3 0 .5 D M S G O -TF ET Position alon g ch an n el (µm) Ban d to Ban d Tu n n elin g R ate (/cm 3 S) 300K 350K 400K 450K (e) V D S = 1.0V P o s itio n a lo ng cha nnel (µm) B a nd to B a nd T unne ling Ra te (/c m 3 S) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K g (f) (e) Fig. 2 Device structure, parameters and simulation details 2 Variation of carrier concentration with temperature for (a) DMSGO-TFET and (b) DMSGO-SDP-TFET, Variation of energy band with temperature for (c) DMSGO-TFET and (d) DMSGO-SDP-TFET, BTBT Variation with temperature for (e) DMSGO-TFET and (f) DMSGO-SDP-TFET. 47 48 49 Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 5 5 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 D r ain c ur r e nt, I D S (A /µm) Fig. 3 Var T hr e shold V oltage , (V ) Fig. 4 (a) and DMSG for the D tional DM SDP-TFE to DMSG tric pocke DMSGO- in Figs. 2 Device structure, parameters and simulation details 3 as ION, I sented in The co with temp convention is shown creases for over, DM cause of r dependen change in DMSGO- ature rang 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 Fig Fig and for tion SDP to D tric DM in F as I sent with con is s crea ove cau dep 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 0 .0 0 .2 0 .4 0 .6 0 .8 1 .0 1 .2 1 0 -1 7 1 0 -1 5 1 0 -1 3 1 0 -1 1 1 0 -9 1 0 -7 1 0 -5 1 0 -3 D r ain c ur r e nt, I D S (A /µm) G a te V o lta g e , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K D M SG O -T FE T V D S = 1 .0 V I OF F Var iation w ith T e m pe r atur e V TH Var iation w ith T e m pe r atur e V TH U sin g C on stan t C u rren t M eth od I ON Var iation w ith T e m pe r atur e (a ) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -17 10 -15 10 -13 10 -11 10 -9 10 -7 10 -5 10 -3 D M S G O -S D P -T F E T V D S = 1.0V D r a in c ur r e nt, I D S (A /µm) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V TH V ariation w ith T emperature I O F F Va ria tio n w ith T em pera ture I O N V ariation w ith T emperature (b ) Fig. 2 Device structure, parameters and simulation details 3 Variation of IDS −VGS characteristics with temperature for (a) conventional DMSGO-TFET, (b) DMSGO-SDP-TFET. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -17 10 -15 10 -13 10 -11 10 -9 10 -7 10 -5 10 -3 D M S G O -S D P -T F E T V D S = 1.0V D r a in c ur r e nt, I D S (A /µm) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V TH V ariation w ith T emperature I O F F Va ria tio n w ith T em pera ture I O N V ariation w ith T emperature (b ) 0 .0 0 .2 0 .4 0 .6 0 .8 1 .0 1 .2 1 0 -1 7 1 0 -1 5 1 0 -1 3 1 0 -1 1 1 0 -9 1 0 -7 1 0 -5 1 0 -3 D r ain c ur r e nt, I D S (A /µm) G a te V o lta g e , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K D M SG O -T FE T V D S = 1 .0 V I OF F Var iation w ith T e m pe r atur e V TH Var iation w ith T e m pe r atur e V TH U sin g C on stan t C u rren t M eth od I ON Var iation w ith T e m pe r atur e (a ) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 g (a ) ta g e , (b ) Fig. 3 Variation of IDS −VGS characteristics with temperature for (a) conventional DMSGO-TFET, (b) DMSGO-SDP-TFET. 16 17 Fig. 2 Device structure, parameters and simulation details 3 Variation of IDS −VGS characteristics with temperature for (a) conventional DMSGO-TF 300K 350K 400K 450K 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55 T e m pe r atur e , (K ) V D S = 1.0 V T hr e shold V oltage , (V ) D M SGO -T FE T D M SGO -SD P-T FE T (a) 300K 350K 400K 450K 20 30 40 50 60 70 80 90 T e m pe r atur e , (K ) V D S = 1.0 V A ve r age SS, (m V /de c ade ) D M SGO -T FE T D M SGO -SD P-T FE T (b) Fig. 4 (a) Comparative threshold voltage and (b) Average subthreshold swing variation with temperature for DMSGO-TFET and DMSGO-SDP-TFET 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 300K 350K 400K 450K 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55 T e m pe r atur e , (K ) V D S = 1.0 V T hr e shold V oltage , (V ) D M SGO -T FE T D M SGO -SD P-T FE T (a) 300K 350K 400K 450K 20 30 40 50 60 70 80 90 T e m pe r atur e , (K ) V D S = 1.0 V A ve r age SS, (m V /de c ade ) D M SGO -T FE T D M SGO -SD P-T FE T (b) (b) Fig. 4 (a) Comparative threshold voltage and (b) Average subthreshold swing variation with temperature for DMSGO-TFET and DMSGO-SDP-TFET 34 35 36 for DMSGO-SDP-TFET compared to DMSGO-TFET. Hence, a substantial increase in drain current is ob- served at lower VGS in the proposed DMSGO-SDP- TFET as shown in the transfer characteristics. Fig. 4(b) illustrates the eﬀect of temperature vari- ation on the average subthreshold swing (SSAV G) for conventional DMSGO-TFET and DMSGO-SDP-TFET. The SSAV G is formulated using the expression [6]. SSAV G = (VT −VOF F ) log(IV T ) −log(IOF F ) (3) (3) The subthreshold swing is inversely proportional to the steepness of IDS −VGS characteristics curve in the sub- threshold region. Fig.4(b) shows that SSAV G for both devices increases as the temperature rises due to an increase in OFF-state current. 2 Device structure, parameters and simulation details Moreover, temperature dependency of DMSGO-SDP-TFET provides 0.28%/K change in SSAV G, whereas it exhibits 0.32%/K change for DMSGO-TFET. Hence, SSAV G variation with tem- perature ranging from 300K to 450K is 0.04%/K lower The subthreshold swing is inversely proportional to the steepness of IDS −VGS characteristics curve in the sub- threshold region. Fig.4(b) shows that SSAV G for both devices increases as the temperature rises due to an increase in OFF-state current. Moreover, temperature dependency of DMSGO-SDP-TFET provides 0.28%/K change in SSAV G, whereas it exhibits 0.32%/K change for DMSGO-TFET. Hence, SSAV G variation with tem- perature ranging from 300K to 450K is 0.04%/K lower 65 Dharmender et al. 2 Device structure, parameters and simulation details 6 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.12 0.14 0.16 0.18 0.20 0.22 (a) D M S G O -T F E T V D S = 1.0V C a pa cita nce, C g s(fF ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.10 0.12 0.14 0.16 0.18 (b) D M S G O -S D P -T F E T V D S = 1.0V C a pa cita nce, C g s(fF ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.0 0.4 0.8 1.2 1.6 2.0 (c ) D M S G O -TF ET V D S = 1 .0 V C a pa cita nce, C g d(fF ) G a te V o lta g e, V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.0 0.4 0.8 1.2 1.6 2.0 (d) D M S G O -S D P -T F E T V D S = 1 .0 V C a pa cita nce, C g d(fF ) G a te V o lta g e, V G S (V) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.00 0.04 0.08 0.12 0.16 0.20 0.24 (e ) D M S G O -T F E T V D S = 1.0 V T r ansc onduc tanc e , g m (m S) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.00 0.20 0.40 0.60 0.80 1.00 (f) D M S G O -S D P -TF ET V D S = 1.0 V T r ansc onduc tanc e , g m (m S) G a te V o lta g e , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K Fig. 2 Device structure, parameters and simulation details 5 Cgs variation with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Cgd Variation with temperature fo (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details gm Variation with temperature for (e) DMSGO-TFET,(f) DMSGO-SDP-TFET 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 F ( f r 3 f T v 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.10 0.12 0.14 0.16 0.18 (b) D M S G O -S D P -T F E T V D S = 1.0V C a pa cita nce, C g s(fF ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.12 0.14 0.16 0.18 0.20 0.22 (a) D M S G O -T F E T V D S = 1.0V C a pa cita nce, C g s(fF ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 (b) (a) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.0 0.4 0.8 1.2 1.6 2.0 (c ) D M S G O -TF ET V D S = 1 .0 V C a pa cita nce, C g d(fF ) G a te V o lta g e, V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.0 0.4 0.8 1.2 1.6 2.0 (d) D M S G O -S D P -T F E T V D S = 1 .0 V C a pa cita nce, C g d(fF ) G a te V o lta g e, V G S (V) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K C a pa cita nce, C g d(fF ) (d) g (c ) g 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.00 0.04 0.08 0.12 0.16 0.20 0.24 (e ) D M S G O -T F E T V D S = 1.0 V T r ansc onduc tanc e , g m (m S) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.00 0.20 0.40 0.60 0.80 1.00 (f) D M S G O -S D P -TF ET V D S = 1.0 V T r ansc onduc tanc e , g m (m S) G a te V o lta g e , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K (f) o g e (e ) g Fig. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 4 8 12 16 20 24 (a) D M S G O -T F E T f T (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K V D S = 1.0V 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 40 80 120 160 200 (b) D M S G O -S D P -TF ET f T (G H Z ) G ate V oltage , V G S (V) 300K 350K 400K 450K V D S = 1.0V (b) (a) Fig. 6 variation of fT with temperature for (a) conventional DMSGO-TFET, (b) DMSGO-SDP-TFET. 16 17 tively. A signiﬁcant decrease in Cgs with increasing tem- perature in both the devices because the potential bar- rier at Source channel interface increases with temper- ature. Similarly, Figs. 5(c) and 5(d) illustrate the Cgd variation for both the devices, respectively. As the ther- mally generated charge carriers in the channel region in- crease the inversion layer. The potential barrier across the drain channel interface decreases as temperature in- creases. Hence, Cgd increases at higher gate bias in both the devices. Also, a similar trend is reported in [29] 19 20 21 22 23 24 25 26 27 28 29 30 Another critical parameter for evaluating the de- vice’s analog/radio frequency performance is the cutoﬀ frequency (fT ). It is deﬁned as the frequency at which current gain becomes 0 dB [28]. It is obtained using the expression fT = gm 2π(Cgs+Cgd). It can be noted that fT depends on Cgs, Cgd and gm of the device. Figs. 6(a) and 6(b) depict the variation in fT with VGS at tem- peratures ranging from 300K to 450K for DMSGO- TFET and DMSGO-SDP-TFET. As seen from Figs. 6(a) and 6(b), fT initially increases with VGS due to an increase in gm and then decreases after a certain value of VGS due to the increased parasitic capacitances and reduced gm because of mobility degradation. 2 Device structure, parameters and simulation details The results also reveal that, with increasing temperature, the peak of fT is shifted more towards lower VGS in DMSGO-SDP-TFET which means that a lower VGS is needed to achieve better high-frequency performance as compared to DMSGO-TFET. Moreover, DMSGO- SDP-TFET exhibits 0.19%/K variation in fT for tem- perature range from 300K to 450K at VGS =0.75 V, whereas for the same biasing conditions and tempera- ture range DMSGO-TFET exhibits 0.39 %/K variation. Hence, the proposed DMSGO-SDP-TFET exhibits bet- ter immunity and reliability to temperature variations for high-frequency applications at higher temperatures when compared to DMSGO-TFET. Transconductance (gm) is one of the most crucial parameters when evaluating a device for its analog/radio frequency and linearity performance. Therefore, higher gm results in achieving higher gain, fT and GBP in the design of analog circuits [31]. In this regard, Figs. 5(e) and 5(f) illustrate the gm variation with temperature for DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. It is evident from the results in the subthreshold region gm of both the devices are very small and it starts increasing due to a steep rise in the ON-state current. However, it starts decreasing after a particu- lar value of VGS because of mobility degradation [32]. The results also reveal that, with increasing tempera- ture, a peak of gm is shifted more towards lower VGS in DMSGO-SDP-TFET which means that a lower VGS is needed to achieve better analog/radio frequency per- formance as compared to DMSGO-TFET. Moreover, DMSGO-SDP-TFET exhibits 0.37%/K variation in gm for temperature ranging from 300K to 450K at VGS = 0.75 V, whereas for the same biasing conditions and temperature range DMSGO-TFET exhibits 0.55 %/K variation. Hence, gm of DMSGO-SDP-TFET exhibits better immunity to temperature variations when com- pared to DMSGO-TFET. This indicates that the pro- posed device is appropriate for analog/radio frequency applications at higher temperatures. 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 The GBP is another important parameter for evalu- ating the analog/radio frequency performance of the de- vice. It is formulated as GBP = gm 20πCgd . The GBP vari- ations for DMSGO-TFET and DMSGO-SDP-TFET with VGS at the temperature range from 300K to 450K, is shown in Figs. 7(a) and 7(b), respectively. 2 Device structure, parameters and simulation details The results also reveal that, with increasing tempera- ture, a peak of gm is shifted more towards lower VGS in DMSGO-SDP-TFET which means that a lower VGS is needed to achieve better analog/radio frequency per- formance as compared to DMSGO-TFET. Moreover, DMSGO-SDP-TFET exhibits 0.37%/K variation in gm for temperature ranging from 300K to 450K at VGS = 0.75 V, whereas for the same biasing conditions and temperature range DMSGO-TFET exhibits 0.55 %/K variation. Hence, gm of DMSGO-SDP-TFET exhibits better immunity to temperature variations when com- pared to DMSGO-TFET. This indicates that the pro- posed device is appropriate for analog/radio frequency applications at higher temperatures. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 Fig. 6 tively. peratu rier at ature. variati mally crease the dr crease the de Tra param freque gm res design and 5 for DM tively. region starts curren lar val The re ture, a in DM is need forman DMSG for tem = 0.75 tempe 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 4 8 12 16 20 24 (a) D M S G O -T F E T f T (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K V D S = 1.0V 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 40 80 120 160 200 (b) D M S G O -S D P -TF ET f T (G H Z ) G ate V oltage , V G S (V) 300K 350K 400K 450K V D S = 1.0V Fig. 6 variation of fT with temperature for (a) conventional DMSGO-TFET, (b) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details 5 Cgs variation with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Cgd Variation with temperature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. gm Variation with temperature for (e) DMSGO-TFET,(f) DMSGO-SDP-TFET. 46 47 48 49 ters such as gm, fT , GBP, fmax,TFP, τ for conventional DMSGO-TFET and the proposed DMSGO-SDP-TFET. The parasitic capacitances (Cgs and Cgd) are cru- cial parameters to analyze the analog/radio frequency and linearity performance of the device. In this regard, Figs. 5(a) and 5(b) illustrate the Cgs variation with VGS at the temperature range from 300K to 450K for DMSGO-TFET and DMSGO-SDP-TFET, respec- Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 7 p y y 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 4 8 12 16 20 24 (a) D M S G O -T F E T f T (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K V D S = 1.0V Fig. 6 variation of fT with temperature for (a) conventional D tively. A signiﬁcant decrease in Cgs with increasing tem- perature in both the devices because the potential bar- rier at Source channel interface increases with temper- ature. Similarly, Figs. 5(c) and 5(d) illustrate the Cgd variation for both the devices, respectively. As the ther- mally generated charge carriers in the channel region in- crease the inversion layer. The potential barrier across the drain channel interface decreases as temperature in- creases. Hence, Cgd increases at higher gate bias in both the devices. Also, a similar trend is reported in [29] Transconductance (gm) is one of the most crucial parameters when evaluating a device for its analog/radio frequency and linearity performance. Therefore, higher gm results in achieving higher gain, fT and GBP in the design of analog circuits [31]. In this regard, Figs. 5(e) and 5(f) illustrate the gm variation with temperature for DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. It is evident from the results in the subthreshold region gm of both the devices are very small and it starts increasing due to a steep rise in the ON-state current. However, it starts decreasing after a particu- lar value of VGS because of mobility degradation [32]. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 Fi at st de at a pa ity ra DM an pr to qu im fm fm 30 TF to th at de 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 5 10 15 20 25 (b) D M S G O -S D P -TF ET V D S = 1.0 V G B P (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 1 2 3 (a) D M S G O -T F E T V D S = 1.0 V G B P (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 (a) g (b) g 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 40 80 120 160 200 (c ) D M S G O -T F E T V D S = 1 .0 V F m a x (G H z) G ate V oltage , V G S (V ) 300K 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 100 200 300 400 500 600 (d) D M S G O -S D P -TF ET V D S = 1.0V F m ax (G H z) G ate V oltage , V G S (V) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K (d) g , (c ) g Fig. 2 Device structure, parameters and simulation details The re- sults reveal that GBP of conventional DMSGO-TFET is inferior to DMSGO-SDP-TFET for the same rea- sons discussed earlier for the fT . Further, it is evident from the results as the temperature increases, GBP 65 Dharmender et al. 8 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 1 2 3 (a) D M S G O -T F E T V D S = 1.0 V G B P (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 5 10 15 20 25 (b) D M S G O -S D P -TF ET V D S = 1.0 V G B P (G H Z ) G ate V oltage , V G S (V ) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 40 80 120 160 200 (c ) D M S G O -T F E T V D S = 1 .0 V F m a x (G H z) G ate V oltage , V G S (V ) 300K 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 100 200 300 400 500 600 (d) D M S G O -S D P -TF ET V D S = 1.0V F m ax (G H z) G ate V oltage , V G S (V) 3 0 0 K 3 5 0 K 4 0 0 K 4 5 0 K Fig. 7 Variation of GBP with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Variation of fmax with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 (b) (a) g (a) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -13 10 -11 10 -9 10 -7 10 -5 10 -3 10 -1 (c ) D M S G O -T F E T V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -14 10 -12 10 -10 10 -8 10 -6 (d) D M S G O -S D P -TF ET V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K 1 T r ansit tim e , (S) (d) g (c ) g Fig. 8 TFP Variation with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Transit time variation with tem- perature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 31 32 33 The transit time (τ) is another important parame- ter for the radio frequency performance assessment of the device. It is deﬁned as the time taken by the de- vice to transfer the carriers from source to drain re- gion [24]. It is formulated as τ = 1 2π10fT . From the above expression, it is evident that transit time de- pends on fT . Figs. 8(c) and 8(d) depict the transit time varies with temperature for DMSGO-TFET and DMSGO-SDP-TFET, respectively. From Fig. 8(c), it can be seen that with increasing VGS, τ decreases in both devices, whereas in the case of DMSGO-SDP- TFET shown in Fig. 8(d) after a certain VGS transit time starts increasing due to decrease in fT because of mobility degradation. Moreover, the results also re- veal that the eﬀect of temperature variations on conven- tional DMSGO-TFET is negligible, whereas DMSGO- SDP-TFET exhibit signiﬁcant variation in transit time. 2 Device structure, parameters and simulation details 7 Variation of GBP with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Variation of fmax with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 1 2 3 mobility degradation started at lower VGS in DMSGO- SDP-TFET which means that a lower VGS is needed to achieve higher fmax as compared to DMSGO-TFET. Therefore, before the mobility degradation i.e, at VGS =0.75 V, and temperature range from 300K to 450K, in terms of %/K variation for fmax, DMSGO-TFET exhibits 0.104 %/K variation and DMSGO-SDP-TFET exhibits 0.075%/K. Hence, the proposed DMSGO-SDP- TFET shows better immunity to temperature varia- tions and exhibits better analog/radio frequency per- formance at lower VGS and elevated temperatures com- pared to DMSGO-TFET. TFP is also an important parameter to consider for the high-frequency performance assessment of the de- vice. It is formulated as TFP = gm IDS fT . Fig. 8(a) and 8(b) depict the TFP variation with temperature for DMSGO-TFET and DMSGO-SDP-TFET, respec- tively. It is evident from the results, for both devices, TFP increases linearly up to a certain value of VGS due to increased gm. After reaching its peak value, it begins to fall due to a decrease in gm and increased parasitic capacitance. 2 Device structure, parameters and simulation details The results also reveal that, as the tem- 65 Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 9 9 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 3 0 6 0 9 0 1 2 0 1 5 0 (a) D M S G O -T F E T T F P (G H Z /V) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V D S = 1.0V 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 2 0 0 4 0 0 6 0 0 8 0 0 1 0 0 0 1 2 0 0 (b) D M S G O -S D P -TF ET T F P (G H Z /V) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V D S = 1.0V 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -13 10 -11 10 -9 10 -7 10 -5 10 -3 10 -1 (c ) D M S G O -T F E T V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -14 10 -12 10 -10 10 -8 10 -6 (d) D M S G O -S D P -TF ET V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K Fig. 8 TFP Variation with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Transit time variation with tem- perature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details 8 TFP Variation with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Transit time variation with tem- perature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. Table 2 Temperature sensitivity in % per kelvin for conven- tional DMSGO-TFET and DMSGO-SDP-TFET. Parameters DMSGO-TFET DMSGO-SDP-TFET VT 0.069 %/K 0.095 %/K SSAV G 0.32 %/K 0.28 %/K ION 1.12 %/K 0.23 %/K gm 0.55 %/K 0.37 %/K fT 0.39 %/K 0.19 %/K GBP 0.27 %/K 0.09 %/K fmax 0.104 %/K 0.075 %/K TFP 0.32 %/K 0.13 %/K perature rises, the TFP of both devices falls due to a decrease in fT and gm because of mobility degradation. Moreover, the results also reveal that at VGS = 0.75 V and temperature range 300K to 450K, in terms of %/K variation, DMSGO-TFET exhibits 0.32 %/K vari- ation in TFP and the proposed DMSGO-SDP-TFET exhibits 0.13%/K. Hence, the proposed DMSGO-SDP- TFET shows better immunity to temperature varia- tions compared to conventional DMSGO-TFET. The transit time (τ) is another important parame- ter for the radio frequency performance assessment of the device. It is deﬁned as the time taken by the de- vice to transfer the carriers from source to drain re- gion [24]. It is formulated as τ = 1 2π10fT . From the above expression, it is evident that transit time de- pends on fT . Figs. 8(c) and 8(d) depict the transit time varies with temperature for DMSGO-TFET and DMSGO-SDP-TFET, respectively. From Fig. 8(c), it can be seen that with increasing VGS, τ decreases in both devices, whereas in the case of DMSGO-SDP- TFET shown in Fig. 8(d) after a certain VGS transit time starts increasing due to decrease in fT because of mobility degradation. Moreover, the results also re- veal that the eﬀect of temperature variations on conven- tional DMSGO-TFET is negligible, whereas DMSGO- SDP-TFET exhibit signiﬁcant variation in transit time. Finally, the temperature sensitivity of diﬀerent device parameters in % per kelvin for conventional DMSGO- TFET and the proposed DMSGO-SDP-TFET are pre- sented in Table 2, and the performance of the proposed device is compared with recent literature in Table 3. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 t V 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 T it ti (S) Fig. 2 Device structure, parameters and simulation details 8 T perature Table 2 tional D Param VT SSAV G ION gm fT GBP fmax TFP peratur decreas Moreov V and %/K va ation in exhibit TFET tions co 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 2 0 0 4 0 0 6 0 0 8 0 0 1 0 0 0 1 2 0 0 (b) D M S G O -S D P -TF ET T F P (G H Z /V) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V D S = 1.0V 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 3 0 6 0 9 0 1 2 0 1 5 0 (a) D M S G O -T F E T T F P (G H Z /V) G a te V o lta g e , V G S (V ) 300K 350K 400K 450K V D S = 1.0V 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 3 0 6 0 (a) T F P (G H G a te V o lta g e , V G S (V ) 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 2 0 0 4 0 0 6 0 0 (b) T F P (G H G a te V o lta g e , V G S (V ) 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -13 10 -11 10 -9 10 -7 10 -5 10 -3 10 -1 (c ) D M S G O -T F E T V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K 0.0 0.2 0.4 0.6 0.8 1.0 1.2 10 -14 10 -12 10 -10 10 -8 10 -6 (d) D M S G O -S D P -TF ET V D S = 1.0 V T r ansit tim e , (S) G ate V oltage , V G S (V ) 300K 350K 400K 450K Fig. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 Fig atu 3.3 dis In on ter DM gar as ysi V I IIP IM plic tor hi 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 0.0 0.2 0.4 0.6 0.8 1.0 1.2 -30 -25 -20 -15 -10 -5 0 5 10 (b) V D S = 1 .0 V g m 3 (m A/V 3 ) G a te V o lta g e ,V G S (V ) 300K 350K 400K 450K D M S G O -S D P -T F E T 0.0 0.2 0.4 0.6 0.8 1.0 1.2 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 2.0 (a) V D S = 1 .0 V g m 3 (m A /V 3 ) G ate V oltage , V G S (V) 300K 350K 400K 450K D M SG O -T FE T (a) (b) g , (a) 0.0 0.3 0.6 0.9 1.2 0 2 4 6 8 10 12 (c ) V D S = 1.0 V D M S G O -T F E T V IP 3 (V ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K (b) 0.0 0.3 0.6 0.9 1.2 0 1 2 3 4 5 6 V DS = 1.0V D M SGO -SD P-T FE T V IP 3 (V ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K (d) g , (d) (c ) Fig. 9 Variation of gm3 with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details Finally, the temperature sensitivity of diﬀerent device parameters in % per kelvin for conventional DMSGO- TFET and the proposed DMSGO-SDP-TFET are pre- sented in Table 2, and the performance of the proposed device is compared with recent literature in Table 3. 65 Dharmender et al. 10 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 0.0 0.2 0.4 0.6 0.8 1.0 1.2 -2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 2.0 (a) V D S = 1 .0 V g m 3 (m A /V 3 ) G ate V oltage , V G S (V) 300K 350K 400K 450K D M SG O -T FE T 0.0 0.2 0.4 0.6 0.8 1.0 1.2 -30 -25 -20 -15 -10 -5 0 5 10 (b) V D S = 1 .0 V g m 3 (m A/V 3 ) G a te V o lta g e ,V G S (V ) 300K 350K 400K 450K D M S G O -S D P -T F E T 0.0 0.3 0.6 0.9 1.2 0 2 4 6 8 10 12 (c ) V D S = 1.0 V D M S G O -T F E T V IP 3 (V ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K 0.0 0.3 0.6 0.9 1.2 0 1 2 3 4 5 6 V DS = 1.0V D M SGO -SD P-T FE T V IP 3 (V ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K (d) Fig. 9 Variation of gm3 with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. Variation of VIP3 with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details It is evi- dent from the results that IIP3 of the proposed DMSGO- Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 11 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 F a T S i e S T p 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 - - - - - IIP 3 (dB m ) -2 -2 -1 -1 - 1 IM D 3 (dB m ) Fig. 10 Va ature for (c Table 3 Co Parame ION (A IOF F (A ION/IO VT H(V) SSavg(m gm(s) fT (GH GBP (G TFP (G SDP-TFET in the IIP3 earity perf SDP-TFET TFET. 2 Device structure, parameters and simulation details Fu perature in are temper A lower IM withstand 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 0.0 0.3 0.6 0.9 1.2 -75 -60 -45 -30 -15 0 15 30 (a) IIP 3 (dB m ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K D M S G O -T F E T V D S = 1 .0 V 0.0 0.3 0.6 0.9 1.2 -75 -60 -45 -30 -15 0 15 30 (b) IIP 3 (dB m ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K D M S GO-S D P -T F E T V D S = 1 .0 V 0.0 0.3 0.6 0.9 1.2 -250 -200 -150 -100 -50 0 50 100 (c ) D M S G O -TF ET V D S = 1 .0 V IM D 3 (dB m ) G ate V oltage , V G S (V) 300K 350K 400K 450K 0.0 0.3 0.6 0.9 1.2 -2 0 0 -1 5 0 -1 0 0 -5 0 0 5 0 1 0 0 (d) D M SGO -SD P-T FE T V D S = 1 .0 V IM D 3 (dB m ) G a te V o lta g e, V G S (V) 300K 350K 400K 450K Fig. 10 Variation of IIP3 with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. IMD3 Variation with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 2 Device structure, parameters and simulation details 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 0.0 0.3 0.6 0.9 1.2 -75 -60 -45 -30 -15 0 15 30 (b) IIP 3 (dB m ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K D M S GO-S D P -T F E T V D S = 1 .0 V 0.0 0.3 0.6 0.9 1.2 -75 -60 -45 -30 -15 0 15 30 (a) IIP 3 (dB m ) G a te V o lta g e, V G S (V ) 300K 350K 400K 450K D M S G O -T F E T V D S = 1 .0 V 0.0 0.3 0.6 0.9 1.2 -2 0 0 -1 5 0 -1 0 0 -5 0 0 5 0 1 0 0 (d) D M SGO -SD P-T FE T V D S = 1 .0 V IM D 3 (dB m ) G a te V o lta g e, V G S (V) 300K 350K 400K 450K 0.0 0.3 0.6 0.9 1.2 -250 -200 -150 -100 -50 0 50 100 (c ) D M S G O -TF ET V D S = 1 .0 V IM D 3 (dB m ) G ate V oltage , V G S (V) 300K 350K 400K 450K (c ) Fig. 10 Variation of IIP3 with temperature for (a) DMSGO-TFET, (b) DMSGO-SDP-TFET. IMD3 Variation with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. 1 2 3 Table 3 Comparison of DC, analog/RF parameters with recent literature Parameters DMSGO-SDP-TFET Ref[33] Ref[34] Ref[35] Ref[23] Ref[36] ION (A/µm) 1.47×10−4 2.06×10−5 1.4×10−4 1.24×10−5 1.21×10−4 1.85×10−5 IOF F (A/µm) 4.56×10−17 2.76×10−17 7.82 ×10−17 5.15×10−18 1.23×10−13 1.26 ×10−16 ION/IOF F 3.14×1012 7.5×1011 1.79 ×1012 2.42×1012 9.83×1011 1.46 ×1011 VT H(V) 0.29 0.31 0.54 0.41 0.41 0.7 SSavg(mV/decade) 29.4 26 34.5 39.8 - 40.44 gm(s) 1.02×10−3 66×10−6 2.61×10−4 6.5×10−5 2.9 ×10−4 7.4×10−5 fT (GHz) 193 46 34 355 59.6 0.17 GBP (GHz) 23.6 7 3.9 36 9.97 0.16 TFP (GHz) 1158 550 - - - 500 SDP-TFET is higher than DMSGO-TFET, this increase in the IIP3 parameter indicates improvement in the lin- earity performance of the device. Therefore, DMSGO- SDP-TFET shows better linearity compared to DMSGO- TFET. 2 Device structure, parameters and simulation details Variation of VIP3 with temper- ature for (c) DMSGO-TFET, (d) DMSGO-SDP-TFET. Figs. 9(a) and 9(b) illustrate the impact of tempera- ture variations ranging from 300K to 450K on higher- order transconductance (gm3) for DMSGO-TFET and DMSGO-SDP-TFET, respectively. It is evident from the above ﬁgures that gm3 variation is negligible at lower VGS for both devices. Moreover, the results also reveal that DMSGO-SDP-TFET is more sensitive to temperature variations at higher gate voltage compared to DMSGO-TFET. In this section, the impact of temperature variations on the linearity and distortion performance parame- ters for conventional DMSGO-TFET and the proposed DMSGO-SDP-TFET have been analyzed. In this re- gard, various linearity and distortion parameters such as gm3, VIP3, IIP3, and IMD3 are considered for anal- ysis. These parameters are deﬁned as follows [31]. 38 39 40 41 42 43 44 45 46 A device with a higher value of VIP3 shows better linearity [25],[31]. Here, Figs. 9(c) and 9(d) illustrate VIP3 variation with VGS at a diﬀerent temperature ranging from 300K to 450K for DMSGO-TFET and DMSGO-SDP-TFET, respectively. It is evident from the results VIP3 of the proposed DMSGO-SDP-TFET is higher and shows better linearity compared to DMSGO- TFET. Further, the results also reveal that both the devices are temperature sensitive for higher VGS. V IP3 = s 24 × gm1 gm3 (4) IIP3 = 2 3 × gm1 gm3 × RS (5) IMD3 = [9 2 × (V IP3)2 × (gm3)]2 × RS (6) 46 47 48 49 50 51 52 53 54 55 V IP3 = s 24 × gm1 gm3 (4) IIP3 = 2 3 × gm1 gm3 × RS (5) IMD3 = [9 2 × (V IP3)2 × (gm3)]2 × RS (6) 46 47 48 49 50 51 52 53 54 55 (4) (5) (6) Where RS is assumed to be 50 Ωfor most RF ap- plications. To obtain better linearity and minimum dis- tortion VIP3, IIP3 parameters of the device should be higher, and IMD3 parameter must be lower [26], [31]. The impact of temperature variations on the IIP3 for DMSGO-TFET and DMSGO-SDP-TFET are illus- trated in Figs. 10(a) and 10(b), respectively. 4 Conclusions IEEE Electron Device Letters 28(8):743-745 (2007) 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 A comparative temperature sensitivity analysis of DC, analog/radio frequency, linearity, and distortion perfor- mance has been carried out for conventional DMSGO- TFET and the proposed DMGOSDG-TFET at the tem- perature ranging from 300K to 450K. Based on this study, it can be stated that for the same biasing con- ditions and temperature range, the DC, analog/radio frequency, and linearity performance parameters of the proposed DMSGO-SDP-TFET are less sensitive to the temperature variation compared to conventional DMSGO TFET. Further, the results summarized in Table 2 in % variation per kelvin, and Table 3 performance compar- ison with recent literature indicate that DMSGO-SDP- TFET is more reliable when compared to DMSGO- TFET. Hence, it can be concluded that the proposed DMSGO-SDP-TFET is a better choice for low power switching and analog/radio frequency applications for elevated temperature range . 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 5. Pal, A., Sachid, A.B., Gossner, H., Rao, V.R.: Insights into the design and optimization of tunnel-FET devices and circuits.IEEE Transactions on Electron Devices 58(4):1045- 1053 (2011) 6. Boucart, K., Ionescu, A.M.: Double gate tunnel FET with high k gate dielectric. IEEE Transactions on Electron Devices 54(7):1725-1733 (2007) 7. Ionescu, A.M., Riel, H.: Tunnel ﬁeld-eﬀect transistors as energy eﬃcient electronic switches. Nature 479(7373):329- 337 (2011) 8. Lee, M.J., Choi, W.Y.: Eﬀects of device geometry on hetero-gate-dielectric tunneling ﬁeld-eﬀect transistors. IEEE Electron Device Letters 33(10):1459-1461 (2012) 9. Jain, P., Prabhat, V., Ghosh, B.: Dual metal-double gate tunnel ﬁeld-eﬀect transistor with mono/hetero dielectric gate material. Journal of Computational Electronics 14(2), 537–542 (2015) 10. Kale, S., Kondekar, P.N.: Charge plasma-based source/drain engineered Schottky Barrier MOSFET: Ambipolar suppression and improvement of the RF per- formance. Superlattices and Microstructures 113: 799-809 (2018) 11. Vijayvargiya, V., Vishvakarma, S.K.: Eﬀect of drain dop- ing proﬁle on double-gate tunnel ﬁeld-eﬀect transistor and its inﬂuence on device RF performance. IEEE Transactions on Nanotechnology 13(5):974-981 (2014) 12. 2 Device structure, parameters and simulation details Further, the results also reveal that as the tem- perature increases from 300K to 450K, both the devices are temperature sensitive at lower and higher VGS. A lower IMD3 parameter indicates that the device can withstand higher distortions [31]. In this regard, Figs. 10(c) and 10(d) illustrate the IMD3 variation with tem- perature. Results demonstrate that DMSGO-SDP-TFET exhibits 0.10%/K variation in IMD3 with temperature ranging from 300K to 450K at VGS = 1.2 V and VDS = 1.0 V, whereas, for the same operating temperature and biasing conditions, DMSGO-TFET exhibits 0.41 %/K variation. Hence, DMSGO-SDP-TFET shows better in- termodulation distortion performance and more reliable to temperature variations as compared to conventional DMSGO-TFET. 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Table 3 Comparison of DC, analog/RF parameters with recent literature Parameters DMSGO-SDP-TFET Ref[33] Ref[34] Ref[35] Ref[23] Ref[36] ION (A/µm) 1.47×10−4 2.06×10−5 1.4×10−4 1.24×10−5 1.21×10−4 1.85×10−5 IOF F (A/µm) 4.56×10−17 2.76×10−17 7.82 ×10−17 5.15×10−18 1.23×10−13 1.26 ×10−16 ION/IOF F 3.14×1012 7.5×1011 1.79 ×1012 2.42×1012 9.83×1011 1.46 ×1011 VT H(V) 0.29 0.31 0.54 0.41 0.41 0.7 SSavg(mV/decade) 29.4 26 34.5 39.8 - 40.44 gm(s) 1.02×10−3 66×10−6 2.61×10−4 6.5×10−5 2.9 ×10−4 7.4×10−5 fT (GHz) 193 46 34 355 59.6 0.17 GBP (GHz) 23.6 7 3.9 36 9.97 0.16 TFP (GHz) 1158 550 - - - 500 34 35 36 37 38 39 40 41 42 43 44 45 46 Table 3 Comparison of DC, analog/RF parameters with recent literature 10(c) and 10(d) illustrate the IMD3 variation with tem- perature. Results demonstrate that DMSGO-SDP-TFET exhibits 0.10%/K variation in IMD3 with temperature ranging from 300K to 450K at VGS = 1.2 V and VDS = 1.0 V, whereas, for the same operating temperature and biasing conditions, DMSGO-TFET exhibits 0.41 %/K variation. Hence, DMSGO-SDP-TFET shows better in- termodulation distortion performance and more reliable to temperature variations as compared to conventional DMSGO-TFET. 65 Dharmender et al. 12 4 Conclusions 4. Saurabh, S., Kumar, M.J.: Novel attributes of a dual ma- terial gate nanoscale tunnel ﬁeld-eﬀect transistor. IEEE Transactions on Electron Devices 58(2): 404-410 (2010) 4 Conclusions A comparative temperature sensitivity analysis of DC, analog/radio frequency, linearity, and distortion perfor- mance has been carried out for conventional DMSGO- TFET and the proposed DMGOSDG-TFET at the tem- perature ranging from 300K to 450K. Based on this study, it can be stated that for the same biasing con- ditions and temperature range, the DC, analog/radio frequency, and linearity performance parameters of the proposed DMSGO-SDP-TFET are less sensitive to the temperature variation compared to conventional DMSGO- TFET. Further, the results summarized in Table 2 in % variation per kelvin, and Table 3 performance compar- ison with recent literature indicate that DMSGO-SDP- TFET is more reliable when compared to DMSGO- TFET. Hence, it can be concluded that the proposed DMSGO-SDP-TFET is a better choice for low power switching and analog/radio frequency applications for elevated temperature range . Declarations Funding: The authors did not receive any ﬁnancial support from any agency/organization for this work. Conﬂict of Interests: No conﬂicts of interest. Availability of data and material (data trans- parency): All the data taken from another resource has been given the corresponding reference. The data, for which reference is not provided, is the original data. Code availability (software application or cus- tom code): The code has been implemented on a 2-D silvaco ATLAS device simulator. Author contributions: Dharmender, Kaushal Nigam, and Satyendra Kumar contributed equally to the de- sign, analysis of the results, and writing of the manuscript. Compliance with ethical standards: The manuscript follows all the ethical standards, including plagiarism. Consent to Participate Yes. Consent for publication: Yes. References 1. Young, K.K.: Short-channel eﬀect in fully-depleted SOI MOSFETs. IEEE Transactions on Electron Devices 36(2):399-402 (1989) 2. Bangsaruntip, S., Cohen, G.M., Majumdar, A., Sleight, J.W.: Universality of short-channel eﬀects in undoped-body silicon nanowire MOSFETs. IEEE Transactions on Electron Devices 31(9):903-905 (2010) 3. Choi, W.Y., Park, B., Lee, J.D., Liu, T.K.: Tunneling ﬁeld-eﬀect transistors (TFETs) with subthreshold swing (SS) less than 60 mV/dec. 4 Conclusions Vivek Anand, I., Arun Samuel, T.S., Vimala, P.: Modeling and simulation of a dual-material asymmetric heterodielectric-gate TFET. Journal of Computational Electronics 19:1450–1462 (2020). https://doi.org/10.1007/s10825-020-01563-w 13. Raad, B.R., Sharma, D., Nigam, K., Kondekar, P.: Group III–V ternary compound semiconductor materials for unipolar conduction in tunnel ﬁeld-eﬀect transistors. Journal of Computational Electronics 16(1): 24-29 (201) 13. Raad, B.R., Sharma, D., Nigam, K., Kondekar, P.: Group III–V ternary compound semiconductor materials for unipolar conduction in tunnel ﬁeld-eﬀect transistors. Journal of Computational Electronics 16(1): 24-29 (201) 14. Kale, S., Kondekar, P.N.: Ferroelectric Schottky barrier tunnel FET with gate-drain underlap: Proposal and investi- gation. Superlattices and Microstructures 89: 225-230 (2016) 14. Kale, S., Kondekar, P.N.: Ferroelectric Schottky barrier tunnel FET with gate-drain underlap: Proposal and investi- gation. Superlattices and Microstructures 89: 225-230 (2016) 15. Kondekar, P.N., Nigam, K., Pandey, S., Sharma, D.: De- sign and analysis of polarity controlled electrically doped tunnel FET with bandgap engineering for analog/RF ap- plications. IEEE Transactions on Electron Devices 64(2):412- 418 (2017) Compliance with ethical standards: The manuscript follows all the ethical standards, including plagiarism. 40 41 Consent to Participate Yes. 42 16. Ashita., Loan, S.A., Rafat, M.: A high-performance inverted- C tunnel junction FET with source-channel overlap pockets. IEEE Transactions on Electron Devices 65(2):763-768 (2018) Consent for publication: Yes. 43 44 ( ) ( ) 17. Kale, S., Kondekar, P.N.: Design and Investigation of Dielectric Engineered Dopant Segregated Schottky Barrier MOSFET With NiSi Source/Drain. IEEE Transactions on Electron Devices 64(11): 4400-4407 (2017) References 48 49 Madan, J., Chaujar, R.: Tem ity issues of heterogeneous ga Tunnel FET. IEEE Transacti 41–48 (2018) 27. Paras, N., Chauhan, S.S.: T ysis of vertical tunneling base analog/RF FOMs. Applied Ph 28. Nigam, K., Pandey, S., K Parte, P.K.: Temperature sen controlled electrostatically do sistor. Superlattices and Micro 29. Singh, P., Yadav, D.S.: Im log/RF, linearity and reliabilit nel FET with ultra-thin source (671) (2020) 30. Kumar, S.: Temperatu logue/RF performance, line tortion for dual-material gate TFET. IET Circuits https://doi.org/10.1049/cds2. 31. Ghosh, P., Haldar, S., Gup vestigation of linearity perfor distortion of GME CGT MOS Transactions on Electron Devic 32. Varshni Y.P.: Temperature in semiconductors. Physica 3 33. Tripathy, M.R., Singh, A. Jit, S.: III-V/Si staggered h pocket engineered vertical TF tions. Superlattice and Microstr 34. Shaikh, M.R.U., S.A.: Loa With Fully Suppressed Amb Application. IEEE Transac 66(4):1628-1634 (2019) 35. Yadav, S., Sharma, D., Son ambipolarity with improved R work function engineering and rial in electrically doped TFET Journal of Computational Elect 36. Seema, Chauhan., S.S.: De tunnel ﬁeld-eﬀect transistor us estimation. Superlattice and M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 21. Dharmender., Nigam, K.: Low-K Dielectric Pocket and Workfunction Engineering for DC and Analog/RF Perfor- mance Improvement in Dual Material Stack Gate Oxide Double Gate TFET. Silicon 13:2347-2356 (2020) 1 2 3 4 Double Gate TFET. Silicon 13:2347-2356 (2020) ( ) 22. Chandan, B.V., Gautami, M., Nigam, K., Sharma, D., Tikkiwal, V.A., Yadav, S., Kumar, S.: Impact of a Metal- strip on a Polarity-based Electrically Doped TFET for Im- provement of DC and Analog/RF Performance. Journal of Computational Electronics 18(1):76-82 (2018) 5 6 7 8 9 ( ) ( ) 23. Raad, B.R., Nigam, K., Sharma, D., Kondekar, P.: Di- electric and work function engineered TFET for ambipolar suppression and RF performance enhancement. Electronics Letters 52(9):770-772 (2016) 9 10 11 12 24. Nigam, K., Kondekar, P.N., Sharma, D.: Approach for ambipolar behavior suppression in tunnel FET by workfunction engineering. IET Micro and Nano Letters 11(8):460-464 (2016) 13 14 15 16 25. References 48 49 1. Young, K.K.: Short-channel eﬀect in fully-depleted SOI MOSFETs. IEEE Transactions on Electron Devices 36(2):399-402 (1989) 50 51 52 18. Rawat, G., Talukdar, J., Mummaneni, K.: A Novel Ex- tended Source TFET with δp+ - SiGe Layer. Silicon 12: 2273–2281 (2020) 19. Kumar, S., Singh, S., Nigam, K., Tikkiwal, V.A.: Dual material dual-oxide dual-gate TFET for improvement in DC characteristics, analog/RF and linearity performance. Applied Physics A 125(5):3531-3538 (2019) 2. Bangsaruntip, S., Cohen, G.M., Majumdar, A., Sleight, J.W.: Universality of short-channel eﬀects in undoped-body silicon nanowire MOSFETs. IEEE Transactions on Electron Devices 31(9):903-905 (2010) 53 54 55 56 20. Joshi, T., Singh, Y., Singh, B.: Extended-Source Double-Gate Tunnel FET With Improved DC and Ana- log/RF Performance. IEEE Transactions on Electron Devices 67(4):1873-1879 (2020) 3. Choi, W.Y., Park, B., Lee, J.D., Liu, T.K.: Tunneling ﬁeld-eﬀect transistors (TFETs) with subthreshold swing (SS) less than 60 mV/dec. IEEE Electron Device Letters 28(8):743-745 (2007) 56 57 58 59 Temperature Sensitivity Analysis of Dual Material Stack Gate Ox 21. Dharmender., Nigam, K.: Low-K Dielectric Pocket and Workfunction Engineering for DC and Analog/RF Perfor- mance Improvement in Dual Material Stack Gate Oxide Double Gate TFET. Silicon 13:2347-2356 (2020) 22. Chandan, B.V., Gautami, M., Nigam, K., Sharma, D., Tikkiwal, V.A., Yadav, S., Kumar, S.: Impact of a Metal- strip on a Polarity-based Electrically Doped TFET for Im- provement of DC and Analog/RF Performance. Journal of Computational Electronics 18(1):76-82 (2018) 23. Raad, B.R., Nigam, K., Sharma, D., Kondekar, P.: Di- electric and work function engineered TFET for ambipolar suppression and RF performance enhancement. Electronics Letters 52(9):770-772 (2016) 24. Nigam, K., Kondekar, P.N., Sharma, D.: Approach for ambipolar behavior suppression in tunnel FET by workfunction engineering. IET Micro and Nano Letters 11(8):460-464 (2016) 25. Nigam, K., Pandey, S., Kondekar, P., Sharma, D., Parte, P.K.: A Barrier Controlled Charge Plasma-Based TFET With Gate Engineering for Ambipolar Suppression and RF/Linearity Performance Improvement. IEEE Transac- tions on Electron Devices 64(6):2751–2757 (2017) 26. Madan, J., Chaujar, R.: Temperature-associated reliabil- ity issues of heterogeneous gate dielectric gate all around Tunnel FET. IEEE Transactions on nanotechnology 17(1): 41–48 (2018) 27. Paras, N., Chauhan, S.S.: Temperature sensitivity anal- ysis of vertical tunneling based dual metal Gate TFET on analog/RF FOMs. Applied Physics A 125: 316 (2019) 28. Nigam, K., Pandey, S., Kondekar, P.N., Sharma, D., Parte, P.K.: Temperature sensitivity analysis of polarity controlled electrostatically doped tunnel ﬁeld-eﬀect tran- sistor. Superlattices and Microstructures 97:598–605 (2016) 29. References 48 49 Nigam, K., Pandey, S., Kondekar, P., Sharma, D., Parte, P.K.: A Barrier Controlled Charge Plasma-Based TFET With Gate Engineering for Ambipolar Suppression and RF/Linearity Performance Improvement. IEEE Transac- tions on Electron Devices 64(6):2751–2757 (2017) 16 17 18 19 20 25. Nigam, K., Pandey, S., Kondekar, P., Sharma, D., Parte, P.K.: A Barrier Controlled Charge Plasma-Based TFET 16 17 18 26. Madan, J., Chaujar, R.: Temperature-associated reliabil- ity issues of heterogeneous gate dielectric gate all around 21 22 26. Madan, J., Chaujar, R.: Temperature-associated reliabil- ity issues of heterogeneous gate dielectric gate all around T l FET IEEE T ti t h l 17(1) 21 22 26. Madan, J., Chaujar, R.: Temperature-associated reliabil- ity issues of heterogeneous gate dielectric gate all around Tunnel FET. IEEE Transactions on nanotechnology 17(1): 41–48 (2018) 21 22 23 26. Madan, J., Chaujar, R.: Temperature associated reliabil ity issues of heterogeneous gate dielectric gate all around ( ) 21 22 Tunnel FET. IEEE Transactions on nanotechnology 17(1): 41–48 (2018) 22 23 24 ( ) 27. Paras, N., Chauhan, S.S.: Temperature sensitivity anal- ysis of vertical tunneling based dual metal Gate TFET on analog/RF FOMs. Applied Physics A 125: 316 (2019) 24 25 26 28. Nigam, K., Pandey, S., Kondekar, P.N., Sharma, D., Parte, P.K.: Temperature sensitivity analysis of polarity controlled electrostatically doped tunnel ﬁeld-eﬀect tran- sistor. Superlattices and Microstructures 97:598–605 (2016) 27 28 29 30 29. Singh, P., Yadav, D.S.: Impact of temperature on ana- log/RF, linearity and reliability performance metrics of tun- nel FET with ultra-thin source region. Applied physics A 127 (671) (2020) 31 32 33 34 30. Kumar, S.: Temperature dependence of ana- logue/RF performance, linearity, and harmonic dis- tortion for dual-material gate-oxide-stack double- gate TFET. IET Circuits Devices & Systems (2021) https://doi.org/10.1049/cds2.12049 35 36 37 38 39 31. Ghosh, P., Haldar, S., Gupta, R.S., Gupta, M.: An in- vestigation of linearity performance and intermodulation distortion of GME CGT MOSFET for RFIC design. IEEE Transactions on Electron Devices 59(12):3263-3268 (2012) 39 40 41 42 Transactions on Electron Devices 59(12):3263-3268 (2012) 42 32. Varshni Y.P.: Temperature dependence of the energy gap in semiconductors. Physica 34(1):149–154 (1967) 43 44 y ( ) ( ) 33. Tripathy, M.R., Singh, A.K., Baral, K., Singh, P.K., Jit, S.: III-V/Si staggered heterojunction-based source- pocket engineered vertical TFETs for low power applica- tions. Superlattice and Microstructures 142:106494 (2020). 44 45 46 47 p 4 ( ) 34. References 48 49 Singh, P., Yadav, D.S.: Impact of temperature on ana- log/RF, linearity and reliability performance metrics of tun- nel FET with ultra-thin source region. Applied physics A 127 (671) (2020) 30. Kumar, S.: Temperature dependence of ana- logue/RF performance, linearity, and harmonic dis- tortion for dual-material gate-oxide-stack double- gate TFET. IET Circuits Devices & Systems (2021) https://doi.org/10.1049/cds2.12049 31. Ghosh, P., Haldar, S., Gupta, R.S., Gupta, M.: An in- vestigation of linearity performance and intermodulation distortion of GME CGT MOSFET for RFIC design. IEEE Transactions on Electron Devices 59(12):3263-3268 (2012) 32. Varshni Y.P.: Temperature dependence of the energy gap in semiconductors. Physica 34(1):149–154 (1967) 33. Tripathy, M.R., Singh, A.K., Baral, K., Singh, P.K., Jit, S.: III-V/Si staggered heterojunction-based source- pocket engineered vertical TFETs for low power applica- tions. Superlattice and Microstructures 142:106494 (2020). 34. Shaikh, M.R.U., S.A.: Loan, Drain-Engineered TFET With Fully Suppressed Ambipolarity for High-Frequency Application. IEEE Transactions on Electron Devices 66(4):1628-1634 (2019) 35. Yadav, S., Sharma, D., Soni, D., Aslam, M.: Controlling ambipolarity with improved RF performance by drain/gate work function engineering and using high K dielectric mate- rial in electrically doped TFET: proposal and optimization. Journal of Computational Electronics 16: 721–731 (2017) 36. Seema, Chauhan., S.S.: Design of double gate vertical tunnel ﬁeld-eﬀect transistor using HDB and its performance estimation. Superlattice and Microstructures 117:1–8 (2018) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 13 Temperature Sensitivity Analysis of Dual Material Stack Gate Oxide Source Dielectric Pocket TFET 21. Dharmender., Nigam, K.: L Workfunction Engineering for mance Improvement in Dual Double Gate TFET. Silicon 1 22. Chandan, B.V., Gautami, Tikkiwal, V.A., Yadav, S., Ku strip on a Polarity-based Elec provement of DC and Analog Computational Electronics 18( 23. Raad, B.R., Nigam, K., Sh electric and work function eng suppression and RF performa Letters 52(9):770-772 (2016) 24. Nigam, K., Kondekar, P. for ambipolar behavior supp workfunction engineering. IE 11(8):460-464 (2016) 25. Nigam, K., Pandey, S., Kon P.K.: A Barrier Controlled C With Gate Engineering for RF/Linearity Performance Im tions on Electron Devices 64(6 26. References 48 49 Shaikh, M.R.U., S.A.: Loan, Drain-Engineered TFET With Fully Suppressed Ambipolarity for High-Frequency Application. IEEE Transactions on Electron Devices 66(4):1628-1634 (2019) 48 49 50 51 ( ) ( ) 35. Yadav, S., Sharma, D., Soni, D., Aslam, M.: Controlling ambipolarity with improved RF performance by drain/gate work function engineering and using high K dielectric mate- rial in electrically doped TFET: proposal and optimization. Journal of Computational Electronics 16: 721–731 (2017) 51 52 53 54 55 ( ) 36. Seema, Chauhan., S.S.: Design of double gate vertical tunnel ﬁeld-eﬀect transistor using HDB and its performance estimation. Superlattice and Microstructures 117:1–8 (2018) 56 57 58	13,754
hal-02739725-2016_Gauffreteau_ISC_1.txt_1	French-Science-Pile	Open Science	Various open science	2,020	Can genotype x environment management interactions (GEMI) be predictedd in sunflower multi-environment trial ?. 19. International Sunflower Conference, May 2016, Edirne, Turkey. ⟨hal-02739725⟩	None	English	Spoken	144	279	Can genotype x environment management interactions (GEMI) be predictedd in sunflower multi-environment trial ? Arnaud Gauffreteau, Margaux d’Orchymont, Celia Pontet, Philippe Debaeke To cite this version: Arnaud Gauffreteau, Margaux d’Orchymont, Celia Pontet, Philippe Debaeke. Can genotype x environment management interactions (GEMI) be predictedd in sunflower multi-environment trial ?. 19. International Sunflower Conference, May 2016, Edirne, Turkey. �hal-02739725� HAL Id: hal-02739725 https://hal.inrae.fr/hal-02739725 Submitted on 2 Jun 2020 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.	13,255
https://openalex.org/W1993737324	OpenAlex	Open Science	CC-By	2,009	How parents choose to use CAM: a systematic review of theoretical models	Ava Lorenc	English	Spoken	11,333	19,600	Published: 22 April 2009 Published: 22 April 2009 BMC Complementary and Alternative Medicine 2009, 9:9 doi:10.1186/1472-6882-9-9 This article is available from: http://www.biomedcentral.com/1472-6882/9/9 This article is available from: http://www.biomedcentral.com This article is available from: http://www.biomedcent © 2009 Lorenc et al; licensee BioMed Central Ltd. © 2009 Lorenc et al; licensee BioMed Central Ltd. © 2009 Lorenc et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creative which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cit Ava Lorenc1, Yael Ilan-Clarke2, Nicola Robinson1 and Mitch Blair3 Ava Lorenc1, Yael Ilan-Clarke2, Nicola Robinson1 and Mitch Address: 1Centre for Complementary Healthcare and Integrated Medicine, Thames Valley University, Paragon House, Boston Manor Road, Brentford, Middlesex, TW8 9GA, UK, 2Lifespan Research Group, Royal Holloway, University of London, 11 Bedford Square, London, WC1B 3RF, UK and 3River Island Paediatric and Child Health Academic Centre, Imperial College, Northwick Park Hospital Campus, Watford Road, Harrow, Middlesex HA1 3UJ, UK Email: Ava Lorenc - ava.lorenc@tvu.ac.uk; Yael Ilan-Clarke - Yael.Ilan-Clarke@rhul.ac.uk; Nicola Robinson - nicky.robinson@tvu.ac.uk; Mitch Blair m blair@imperial ac uk Email: Ava Lorenc - ava.lorenc@tvu.ac.uk; Yael Ilan-Clarke - Yael.Ilan-Clarke@rhul.ac.uk; Nicola Robinson - nicky.robinson@tvu.ac.uk; Mitch Blair* - m.blair@imperial.ac.uk * Corresponding author Received: 1 September 2008 Accepted: 22 April 2009 Received: 1 September 2008 Accepted: 22 April 2009 Received: 1 September 2008 Accepted: 22 April 2009 BMC Complementary and Alternative Medicine BioMed Central BioMed Central Open Access Abstract Background: Complementary and Alternative Medicine (CAM) is widely used throughout the UK and the Western world. CAM is commonly used for children and the decision-making process to use CAM is affected by numerous factors. Most research on CAM use lacks a theoretical framework and is largely based on bivariate statistics. The aim of this review was to identify a conceptual model which could be used to explain the decision-making process in parental choice of CAM. Methods: A systematic search of the literature was carried out. A two-stage selection process with predetermined inclusion/exclusion criteria identified studies using a theoretical framework depicting the interaction of psychological factors involved in the CAM decision process. Papers were critically appraised and findings summarised. Results: Twenty two studies using a theoretical model to predict CAM use were included in the final review; only one examined child use. Seven different models were identified. The most commonly used and successful model was Andersen's Sociobehavioural Model (SBM). Two papers proposed modifications to the SBM for CAM use. Six qualitative studies developed their own model. Conclusion: The SBM modified for CAM use, which incorporates both psychological and pragmatic determinants, was identified as the best conceptual model of CAM use. This model provides a valuable framework for future research, and could be used to explain child CAM use. An understanding of the decision making process is crucial in promoting shared decision making between healthcare practitioners and parents and could inform service delivery, guidance and policy. http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 recent definitions divides CAM into mind-body medicine, biologically based therapies, manipulative and body- based systems and energy medicine, and whole system approaches such as Ayurveda and Traditional Chinese Medicine[5]. readily and taking caution in using home remedies[25]. However, they may also feel more strongly dissatisfied with conventional healthcare for their child than for themselves[29]. In addition, females are higher users of healthcare service[30], and mothers are more likely to use CAM for their children than fathers[31,32], indicating that mainly mothers are involved in decision making. CAM is very popular, with recent population based esti- mates of yearly adult use in the UK of 20% – 28%[6,7] and 34% – 38% in the USA [8,9]. A systematic review of CAM prevalence surveys worldwide found a prevalence of between 23% – 62%, and for over the counter CAM 25% – 46%[10]. CAM is also commonly used for children, with prevalence estimates from 12% in the USA[9], 11% in Canada[11] to 51% in Australia[12]and 17.9%[13] to 37%[14] in the UK. There is evidence that home remedies are also commonly used for children in the UK[13,15,16]. There is a large amount of literature written on the deci- sion-making process in conventional care but the extrap- olation of the conventional medicine (CM) decision- making process to CAM choices is debatable[33]. The process of choosing to use CAM may be far more dynamic, iterative and more individualistic than the more logical and rational decisions in conventional care[33,34]. As in making choices about conventional care treatments, in choosing CAM there are numerous considerations to take into account. Such considerations revolve around the personal perception of the balance between expected drawbacks of action (e.g. side effects) and the anticipated benefits of the treatment[17]. Within this vast continuum, the variables which will impact on the decision making process include desires (utilities associated with each alternative, personal values, goals, etc), beliefs (expecta- tions about processes and outcome, knowledge, means to achieve desired outcome etc)[18] and other practical con- siderations (e.g. access). Decision making in healthcare may be moving from paternalism to autonomy, finally settling on shared decision making and 'consumer- ism'[19]. The literature about choosing to use CAM encompasses many different approaches. Many studies have identified the factors associated with CAM use in children in the UK[13,15,35,36], the USA[32,37-42] and Canada[11,43]. http://www.biomedcentral.com/1472-6882/9/9 However, much of the research into reasons for using CAM so far has been atheoretical and lacks a clear and comprehensive conceptual framework to contain and explain the processes which are inherent in CAM decision- making[44,45]. Most of the studies are based on survey methods, are cross-sectional and cannot determine direc- tional relationships[46]. In recent years there has been a move to explain the mechanisms which motivate and actualise the choice of CAM. This review examines the literature relating to the deci- sion-making process in CAM, but excludes models which do not include psychosocial factors or affective values or beliefs such as computational models of decision making, the cognitive processes involved in decision making (e.g. Hypothetico-deductive model[47]) and other descriptive theories of the decision making process which relate to treatment choices (e.g. Prospect theory[48]). This review focuses on the decision which leads to choosing CAM; as such it concerns itself with the models which attempt to explain the choice of complementary or alternative healthcare and the psychosocial factors that are involved in this decision. In addition, the decision making process for child health is different to that of adults as it may include the whole family not just the patient[20]. Decisions are often made by parents, not the child although there is debate around the role of children, with the model of constrained paren- tal autonomy suggesting that parents make decisions for their children but this autonomy is not absolute[21]. Family centred care is now a central tenet of healthcare, particularly nursing[22], although a recent literature review found that despite the importance of including children in decision making on their own health they are rarely involved in the decision making process[23]. Regarding CAM use, particularly for certain ethnic minor- ity groups, children may be even less autonomous than in other areas[24,25]. Adolescents however are likely to have a greater degree of autonomy, using CAM due to personal beliefs and control[26] Parents may use CAM to be a 'good' parent, particularly for children with a serious ill- ness[27]. In addition parents may not use the same treat- ments for children as for themselves, including home remedies[28]. Background definitions varying over time[4]. CAM can be defined as "any health improving technique outside of the main- stream of conventional medicine"[2]. One of the most Background There is considerable debate around the definition of Complementary and Alternative Medicine (CAM) [1-3], Page 1 of 12 (page number not for citation purposes) Page 1 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 Page 2 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 Parents may be more cautious with chil- dren's health, trusting and visiting practitioners more Two dominant approaches have been used to study deci- sion-making in CAM: the first originates in the concept of healthcare utilisation, concerned with the factors which enable and encourage the consumption of health services. The second approach views the decision to use CAM as a health behaviour, where the decision to use CAM is viewed within the framework of social and psychological, mainly cognitive, factors. Background on the models reviewed is given in table 1. Page 2 of 12 (page number not for citation purposes) Page 2 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 BMC Complementary and Alternative Medicine 2009, 9:9 Table 1: Descriptions of the models Model Description References Andersen's Sociobehavioural Model (SBM) Sets out three sequential components which mitigate healthcare use. The first most indirect, are predisposing factors including beliefs, sociodemographics and characteristics which motivate the healthcare service use. The next component, more directly related to behaviour, are enabling factors which allow and give access to healthcare services (e.g. income, physical location, insurance). The final most proximal component is medical need, including the objective and subjective experience of symptoms of illness [65] Consumer Decision-Making model (CDM) Has three components: first is external influences: sociocultural influences on beliefs, knowledge and behaviours. Second is the consumer decision-making process; including psychological influences (values, beliefs, attitudes, personality) which form the main part of the decision making process. Finally, the post decision behaviour consists of the behaviour itself and an evaluative comparison of the actual experience with the anticipated experience. [51] Health locus of control (HLoC) The HLoC identifies the extent to which people perceive their health, treatment, course of illness and other health related factors, to be under their control or external to them (e.g. fate, doctor, others). [55,61] Transtheoretical model (TTM) TTM engenders five stages: 1. Precontemplation (no intention to make change) 2. Contemplation (consideration of making change) 3. Preparation (effecting small steps to begin change) 4. Action (carrying out the change to its full extent) 5. Maintenance (sustaining the change over time). Healthcare utilisation models The decision leading to choice of healthcare can be mod- elled by pathway (sequential), or determinants mod- els[49]. Pathway models give stages of healthcare seeking, moving from self-care, adoption of the sick role, seeking medical care and finally recovery[49]. Determinants mod- els focus on explanatory factors of the choices made[49]. The Consumer Decision-Making model which is less often used for healthcare has three components: external influences, the consumer decision-making process and the post decision behaviour[51]. The main determinants model is Andersen's sociobehav- ioural model (SBM)[30,50], particularly prominent in the (conventional) medicine literature[30]. This model sets out three sequential components which mitigate health- care use; predisposing, enabling and need factors[30]. A recent addition to the model is the role of social support, whereby social influence can encourage the utilisation of The majority of healthcare utilisation models have found that the dominant determinant of healthcare utilisation is need, or illness[49]. http://www.biomedcentral.com/1472-6882/9/9 [72] Health locus of control (HLoC) Transtheoretical model (TTM) Health locus of control (HLoC) The HLoC identifies the extent to which people perceive their health, treatment, course of illness and other health related factors, to be under their control or external to them (e.g. fate, doctor, others). [55,61] [92] 1. Precontemplation (no intention to make change) 2. Contemplation (consideration of making change) 3. Preparation (effecting small steps to begin change) 4. Action (carrying out the change to its full extent) 5. Maintenance (sustaining the change over time). The distinguishing characteristics of this model are firstly that moving through the stages is not necessarily a linear process, but it is necessary to move through all changes in order to incur sustained change; Secondly, the balance of pros and cons of carrying out a given behaviour, will determine the stage of change in which the individual finds him/her self. [57]. Theory of planned behaviour (TPB) TPB attempts to explain behavioural intentions as predicted from three major sources: attitudes, perceived behavioural control and subjective norms. Attitudes include beliefs and expectations about a particular behaviour and the extent to which consequences are seen as desirable. Subjective norms are the beliefs one has about the expectations of 'significant others' and the motivation to comply with these. Perceived behavioural control is the extent to which one expects the behaviour to be easy or difficult and whether they perceive themselves to have the ability to carry out such a behaviour – often equated with self efficacy [5 The SRM explains how individuals have 'illness beliefs' or 'illness perceptions' about their condition. These are predefined cognitions which represent illness characteristics and coping strategies, related to perceived cause, effects, consequences, duration and sources of control or cure. People go on to form a representation of their coping alternatives, which may be represented as 'treatment beliefs'. [70,93] Braden's self-help model specifies central variables and relationships involved in a learned response to chronic illness and includes the following elements: side-effects burden, uncertainty, perceived enabling skills, self help and quality of life. Its utility lies in its ability to form the connection between individual's use of enabling skills to manage their illness. [72] healthcare as well as the perception of the efficacy of a given treatment[50]. http://www.biomedcentral.com/1472-6882/9/9 The distinguishing characteristics of this model are firstly that moving through the stages is not necessarily a linear process, but it is necessary to move through all changes in order to incur sustained change; Secondly, the balance of pros and cons of carrying out a given behaviour, will determine the stage of change in which the individual finds him/her self. [92] Theory of planned behaviour (TPB) TPB attempts to explain behavioural intentions as predicted from three major sources: attitudes, perceived behavioural control and subjective norms. Attitudes include beliefs and expectations about a particular behaviour and the extent to which consequences are seen as desirable. Subjective norms are the beliefs one has about the expectations of 'significant others' and the motivation to comply with these. Perceived behavioural control is the extent to which one expects the behaviour to be easy or difficult and whether they perceive themselves to have the ability to carry out such a behaviour – often equated with self efficacy [57]. The self-regulatory model (SRM) The SRM explains how individuals have 'illness beliefs' or 'illness perceptions' about their condition. These are predefined cognitions which represent illness characteristics and coping strategies, related to perceived cause, effects, consequences, duration and sources of control or cure. People go on to form a representation of their coping alternatives, which may be represented as 'treatment beliefs'. [70,93] Braden's Self-help model Braden's self-help model specifies central variables and relationships involved in a learned response to chronic illness and includes the following elements: side-effects burden, uncertainty, perceived enabling skills, self help and quality of life. Its utility lies in its ability to form the connection between individual's use of enabling skills to manage their illness. [72] Table 1: Descriptions of the models Model Description Andersen's Sociobehavioural Model (SBM) Sets out three sequential compo indirect, are predisposing factors which motivate the healthcare se to behaviour, are enabling factor income, physical location, insura need, including the objective and Consumer Decision-Making model (CDM) Has three components: first is ex knowledge and behaviours. Seco psychological influences (values, of the decision making process. behaviour itself and an evaluative anticipated experience. Health locus of control (HLoC) The HLoC identifies the extent t of illness and other health relate (e.g. fate, doctor, others). Transtheoretical model (TTM) TTM engenders five stages: 1. Precontemplation (no intentio 2. Contemplation (consideration 3. Preparation (effecting small st 4. Action (carrying out the chang 5. http://www.biomedcentral.com/1472-6882/9/9 Finally, the post decision behaviour consists of the behaviour itself and an evaluative comparison of the actual experience with the anticipated experience. [51] The HLoC identifies the extent to which people perceive their health, treatment, course of illness and other health related factors, to be under their control or external to them (e.g. fate, doctor, others). [55,61] TTM engenders five stages: 1. Precontemplation (no intention to make change) 2. Contemplation (consideration of making change) 3. Preparation (effecting small steps to begin change) 4. Action (carrying out the change to its full extent) 5. Maintenance (sustaining the change over time). The distinguishing characteristics of this model are firstly that moving through the stages is not necessarily a linear process, but it is necessary to move through all changes in order to incur sustained change; Secondly, the balance of pros and cons of carrying out a given behaviour, will determine the stage of change in which the individual finds him/her self. [92] TPB attempts to explain behavioural intentions as predicted from three major sources: attitudes, perceived behavioural control and subjective norms. Attitudes include beliefs and expectations about a particular behaviour and the extent to which consequences are seen as desirable. Subjective norms are the beliefs one has about the expectations of 'significant others' and the motivation to comply with these. Perceived behavioural control is the extent to which one expects the behaviour to be easy or difficult and whether they perceive themselves to have the ability to carry out such a behaviour – often equated with self efficacy [57]. The SRM explains how individuals have 'illness beliefs' or 'illness perceptions' about their condition. These are predefined cognitions which represent illness characteristics and coping strategies, related to perceived cause, effects, consequences, duration and sources of control or cure. People go on to form a representation of their coping alternatives, which may be represented as 'treatment beliefs'. [70,93] Braden's self-help model specifies central variables and relationships involved in a learned response to chronic illness and includes the following elements: side-effects burden, uncertainty, perceived enabling skills, self help and quality of life. Its utility lies in its ability to form the connection between individual's use of enabling skills to manage their illness. http://www.biomedcentral.com/1472-6882/9/9 Maintenance (sustaining the ch The distinguishing characteristics is not necessarily a linear proces order to incur sustained change; given behaviour, will determine t self. Theory of planned behaviour (TPB) TPB attempts to explain behavio attitudes, perceived behavioural and expectations about a particu seen as desirable. Subjective nor 'significant others' and the motiv control is the extent to which o whether they perceive themselv often equated with self efficacy The self-regulatory model (SRM) The SRM explains how individua condition. These are predefined coping strategies, related to perc sources of control or cure. Peop alternatives, which may be repre Braden's Self-help model Braden's self-help model specifie response to chronic illness and i uncertainty, perceived enabling s to form the connection between odels Description References del (SBM) Sets out three sequential components which mitigate healthcare use. The first most indirect, are predisposing factors including beliefs, sociodemographics and characteristics which motivate the healthcare service use. The next component, more directly related to behaviour, are enabling factors which allow and give access to healthcare services (e.g. income, physical location, insurance). The final most proximal component is medical need, including the objective and subjective experience of symptoms of illness [65] el (CDM) Has three components: first is external influences: sociocultural influences on beliefs, knowledge and behaviours. Second is the consumer decision-making process; including psychological influences (values, beliefs, attitudes, personality) which form the main part of the decision making process. Finally, the post decision behaviour consists of the behaviour itself and an evaluative comparison of the actual experience with the anticipated experience. [51] healthcare as well as the perception of the efficacy of a els Description References (SBM) Sets out three sequential components which mitigate healthcare use. The first most indirect, are predisposing factors including beliefs, sociodemographics and characteristics which motivate the healthcare service use. The next component, more directly related to behaviour, are enabling factors which allow and give access to healthcare services (e.g. income, physical location, insurance). The final most proximal component is medical need, including the objective and subjective experience of symptoms of illness [65] CDM) Has three components: first is external influences: sociocultural influences on beliefs, knowledge and behaviours. Second is the consumer decision-making process; including psychological influences (values, beliefs, attitudes, personality) which form the main part of the decision making process. Health behaviour models Health behaviour models take into account psychological influences on behaviour and thus explain the individual Page 3 of 12 (page number not for citation purposes) Page 3 of 12 (page number not for citation purposes) BMC Complementary and Alternative Medicine 2009, 9:9 http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 Two stages of screening were used to identify relevant arti- cles. A diagram of the process of inclusion/exclusion is given in Figure 1. differences in behaviour, but often do not take account of external characteristics such as sociodemographic varia- bles. Within this approach there are various models which have been applied to health behaviours, such as the Health locus of Control (HLoC), the Theory of Planned Behaviour (TPB), the Transtheoretical model (Stages of change) (TTM) and the Self-Regulatory model (SRM). At first screening exclusion criteria were: randomised con- trol trial/efficacy trials of CAM; physician/practitioner knowledge/choice/decision making; teaching in medical school; integration into primary care; non-English lan- guage; relating to regulation of CAM; survey, comment, editorial or review; published before the year 1995. The concept of HLoC originates in Attribution Theory [52] and relates to the extent to which individuals view events as under their control (an internal locus of control) or out of their control (external locus of control). HLoC has been widely applied in the health and other arenas [53,54], based on the prediction that individuals high in internal locus of control are more likely to carry out health-promoting behaviours, whereas those with high external locus of control who attribute their health to chance will be unlikely to engage in health-enhancing activities [55]. The TTM was originally developed as a charting of the processes engendered in the elicitation and maintenance of change during the therapeutic process [56] and has been used for various health behaviours. The theory of planned behaviour[57] attempts to explain behavioural intentions as predicted from three major sources: attitudes, perceived behavioural control and sub- jective norms. The SRM was formulated to describe the process whereby when confronted with a health threat, individuals seek overcome the problem and return to nor- mality. At the second selection stage the inclusion criteria were that the paper had to present a model of factors associated with the utilisation or choice of CAM. Results Over 2700 articles were screened for inclusion. Only one study focussed on children, so the review had to include articles related to adult decision making. As seen in Figure 1, 22 articles met the criteria for inclusion in the review. These studies are presented in Additional file 1, and dis- cussed below. This paper presents the results of a systematic review to identify how these models have been applied to the pre- diction of CAM use, and focuses on their potential use for children. The paper discusses the methods used in the lit- erature search, followed by the results which discuss the quality of the papers found and their conclusions regard- ing the suitability of the model they test. Methods Aim To identify a conceptual framework which can success- fully model the parental decision making process of choosing to use CAM for children, through a systematic review of studies using a decision making model for CAM use. Health behaviour models This was done by screening each paper for the key word 'model' or 'theory' and determining their use or reference to a model within the article (excluding the term 'regression model' and 'integrative model/integrative care model'). The term model was occasionally used in reference to a paradigm, rather than the intended use as in a theoretical framework which depicts a process of sorts; when this occurred it was necessary to exclude such studies. Models h In the current review 7 established models and four 'uni- dentified' models were used to predict the psychological process of choice of CAM: scales relating to the locus of control [58-62], TPB [63], TTM [63], Andersen's sociobe- havioural model [29,58,64-69], the self-regulatory model [70,71], the consumer decision making model [44] and Braden's self-help model [72]. Six studies [71,73-77] con- structed models based on their findings, including one using ethnographic decision tree modelling[71]. Only one study focussed on child use of CAM[29]. Measures and analyses tion of a large amount of variables. This often resulted in largely correlational datasets, which, while being inform- ative and to some extent predictive, fail to provide a causal model of CAM decision-making and to identify the exact mechanisms through which CAM choices are made. Only one study examined the propensity for CAM use over two time points, thereby assigning specific causal relation- ships to use of CAM[67]. Most of the quantitative studies included in this review tended to use measures which were largely found to be reliable and valid. This was often established in previous studies which used the same measures. In some cases, the reliability of the measures was tested through internal consistency (Cronbach ά) and multi-item responses used to establish reliability within the studies. Aside from the qualitative studies and a number of the surveys, many of the studies were self-report questionnaires, which are open to biases in the form of response bias, demand char- acteristics and to the introduction of systematic errors. One major advantage of a number of studies was that they used the frequency of CAM use as a variable to compare specific predictors of more or less use. By doing this they were able to distinguish between the beliefs which differ- entiated a person who tried CAM but did not have a com- mitted treatment plan [58]. The qualitative studies used semi structured or open ended interviews. They also tended to use methods which attest to the integrity and validity of the data such as con- firming the findings with participants. One qualitative study tested the predictability of the model they devel- oped[71]. Design Out of 22 studies surveyed, 16 used quantitative approaches to examine the models in question. Six studies [71,73-77] used qualitative methodology. In the initial search, many studies were identified which used qualita- tive methodology to examine the subject of predictors or correlates of CAM use, however in the subsequent analysis these had to be excluded due to lack of use of a model. A systematic search was conducted of the following data- bases: Psychinfo, Sciencedirect, Academic search elite, Medline, Psycharticles, Elsevier, Biomed, Ingenta connect, Cinahl and Embase. A combination of the following search terms was used: CAM or Complementary or alter- native Medicine, choice, decision making, parent or child or adolescent or paediatric or pediatric, model, utiliz. All the studies, excluding one [67] used a cross-sectional design. The main disadvantage of such a design is that cause and effect cannot be determined in spite of collec- Page 4 of 12 (page number not for citation purposes) Page 4 of 12 (page number not for citation purposes) BMC Complementary and Alternative Medicine 2009, 9:9 http://www.biomedcentral.com/1472-6882/9/9 wchart of article selection process gure 1 owchart of article selection process. 459 Over 2700 papers identified in original search Selected for relevance Relevance to CAM use n =133 326 Duplicates excluded n =48 278 Not English language: n =15 263 Related to Health professionals / integrative model n = 30 233 Focus on RCT /efficacy trials N = 12 221 Editorial / review / comments N = 29 193 187 177 Pre 1995 N= 6 Only prevalence / no identifiable model N = 154 Not available (eg dissertation) N= 10 N = 22 articles included in the review Over 2700 papers identified in original search Selected for relevance Relevance to CAM use n =133 Duplicates excluded n =48 Not English language: n =15 Related to Health professionals / integrative model n = 30 Focus on RCT /efficacy trials N = 12 Editorial / review / comments N = 29 Pre 1995 N= 6 Not available (eg dissertation) N= 10 Only prevalence / no identifiable model N = 154 N = 22 articles included in the review Flowchart of article selection process Figure 1 Flowchart of article selection process. Page 5 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 Limitations of the studies Many studies did not distinguish between different types of CAM, which may have significant implications given that those studies that did differentiate CAM type found that the decision making process did vary for the different CAM modalities[66,68-70]. Some studies were unclear about what was included in their definition of CAM[67,72]. Overall, the response rate for most studies was highly acceptable at 60% or higher for most studies[82]. This serves to enhance the reliability of the findings in terms of their generalisability. However, for a few studies there was very low response rate, which would indicate that it was unlikely that the sample was representative of the popula- tion concerned[44,60,62]. Not all studies controlled for factors which may have biased the sample or introduced extraneous variables, such as; stage of illness, duration of illness and conven- tional or other treatments[61,67,72]. Studies using the SBM did not always explain the recursive nature of the fac- tors which has recently been described[50]. Most studies only included adults, most defined as over 18, but some with limited age ranges[65,70,71,76]. Only one paper [29] focused on the use of CAM for children and utilized a theoretical model. Although some studies were based on large, nationally representative samples, some used small, potentially underpowered samples[58,59,62]. Two studies addition- ally only included CAM users, preventing comparison of CAM users and non users[58,75]. Although most studies did not specifically exclude non CAM users, there may have been response bias in terms of CAM users being more likely to take part. Sampling The majority (10) of studies surveyed had medium sized samples of between 123 and 551, indicating attempts to achieve a representative population. Of the studies using self-selection there was a predominance of female respondents, usually because females were more likely to use health services [78] and more likely to use or consider using CAM [6,8,79-81]. Some studies were specifically aimed at the experience of females only, mainly in rela- tion to breast cancer and other female-dominated diseases [61,64,72,73,75], one included males only (prostate can- cer)[76]. Other studies based on large scale, often national, studies had much larger samples of between 1672 and 31,044 so were able to pursue a more represent- ative sample in most cases. Qualitative studies had much smaller samples, between 16 and 42, which is appropriate for their methodology. The quantitative studies predominately used multivariate logistic regression or multinomial logit regression to explain the relative variance of each of the factors signifi- cant in the decision making process although some anal- yses were limited to bivariate or correlational association[59,66]. The qualitative studies used either grounded theory[73,75], or 'thematic' analysis[74,76,77]. CAM definition In the studies included in this review not all studies made the distinction between different types of CAM therapies. Hendrickson et al highlight the problem of treating CAM as one modality and illustrate through their study that there are differences in the determinants of use of differ- ent type of CAM therapies[68]. Most studies included only practitioner based therapies, others viewed CAM in terms of the behaviour. The lack of a consistent opera- tional definition of CAM use made the papers heterogene- ous and difficult to combine and form conclusions and may explain some of the inconsistent findings in the liter- ature. There is a need to examine studies which identify types of CAM in order to compare their findings, reflected in Andersen's suggestion that the outcome of the SBM The findings from the studies using the SBM are summa- rised in Figure 2. As described by Andersen[50], the Factors in the SBM important for CAM use Figure 2 Factors in the SBM important for CAM use. References show which factors were important in which studies. ! " # # $ % & $ $ ' ! ( ) ( " # # # $ ' " + $ $ , - " , - " . ( * " / # & " 0 " . 1 2 # # ! CAM definition " . 2 & 2 ' " . 2 # 3 # # % & 4 * " 5 6 7 8 6 9 : : : ; < < = > ? @ = > ? A 9 : 9 @ B C ? = > : D E $ F # # G ' ! ( ' ( ! ( * " / # ! ( * " # ' " % 2 & - " ' " & D 2 ' " H 7 H 7 8 0 $ # F $ 2 G - ( ' Ͳ ! " I F 2 $ G 4 * ( ) ( ' ( ! http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 should ideally relate to a specific type of healthcare serv- ice[50]. In addition, a distinction between CAM use as a treat, a preventative strategy or treatment of disease was often lacking; This may serve to distinguish between diverse CAM users who have different motivations for using such services[66] In addition other factors influenc- ing the status of CAM which may affect the decision mak- ing process will vary between, and even within, countries of study. These include professional regulation, legal sta- tus, financial access and reimbursement of CAM and its integration within national health systems. son between different studies difficult. In particular a number of the studies based on the SBM used non vali- dated measures of health beliefs [65,66], or did not include health beliefs at all[68,69]. Some studies did not provide statistics on the percentage of variance the model explained[59,66]. Healthcare utilisation models The socio-behavioural model was the most commonly used, and was largely supported for modelling CAM deci- sion making, although some studies were only partially supportive of the model; most commonly enabling fac- tors were not significant. One study[69] was based on an adapted SBM model, the "CAM Healthcare Model"[83]. Here the SBM was extended to examine the concurrent, complementary use of conventional medicine and CAM, and the choice between them, and included self-care prac- tices and products as well as practitioner based CAM[83]. The only study using the SBM for child CAM use added the component of healthcare experience[29]. These adap- tations may be important for child use of CAM which is often non practitioner based (88% of CAM use by London paediatric outpatients[84] and 64% in the USA was non practitioner based[85]), and parental use of CAM is very likely to influence child use[11,13,29,40,86,87]. Discussion h The current review found that almost 100 papers (elimi- nated from the final analysis) did not use an overarching framework to examine their findings, leaving them open to spurious explanations. Some studies investigated psy- chological constructs such as beliefs, using validated measures, but refrained from going further to consolidate a model. Other studies set out to validate the items and their interrelationships within a model, discussed below (see table 1 for descriptions of models). CAM definition ( * " F # # G - Ͳ * " D F G - ( ) ( ! ( * " ' " J 2 K 1 & ) ( ' " L ) " / # 2 2 F $ $ $ G ) " . $ % # ' " 1 2 ' " E 2 ' " M N O P Q R S 6 S T H 7 . 2 & $ 2 # 2 4 * " 0 2 U # $ ' ! " 5 6 7 8 6 9 : : : ; < < = > ? @ = > ? A 9 : 9 @ B C ? = > : S 6 S T H 7 . Settings Studies were mainly conducted in the US, Canada, Japan and the UK (although limited by English language inclu- sion only). Participants were recruited from conventional medicine (CM) centres, CAM clinics, health related inter- net sites, national surveys or random internet mailing. A number of studies used non validated measures, which limited the validity of the study and also makes compari- Page 6 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 Health behaviour models This review only identified one study using a theoretical model for child use of CAM. Although this study found support for most components of the SBM, this study was based in the USA, only tested practitioner based CAM, and the survey was not specifically designed to test the SBM. The lack of studies on child use of CAM using a theoretical model means this review is unable to extrapolate findings to the use of CAM in children. However it does highlight the need to identify the most suitable model and to test it's suitability for application to CAM use both for parents and children to determine whether the processes are sim- ilar to the choices of CAM in adults. The original SBM used the family as the unit of study[30], and although recently this has changed to the individual, due to prob- lems in measuring family based variables[50], the family focus is especially appropriate for child CAM use. When CAM use is viewed as a health behaviour, individ- ual differences which are not explainable in terms of more extrinsic characteristics (e.g. socio-demographic) can be explained. This is important given that CAM use is often a behaviour specific to an individual[77]. The advantages of this approach are two-fold; Firstly psychological factors, such as cognition, beliefs and values, are considered to be important and proximal to the decision to carry out cer- tain behaviour, and may mediate other more extrinsic fac- tors[88]. Secondly, psychological factors, as opposed to extrinsic factors, have the advantage of being amenable to change, at least to some extent; this is particularly impor- tant in relation to health-related interventions including CAM[88]. The health locus of control was predictive of CAM use in two studies[59,61] but not in three[58,60,62]. In addi- tion, many of the HLoC studies were carried out on sam- ples of patients with a chronic illness, so of limited generalisability. In terms of the decision to use CAM, it can relate to the perception of control over illness and treatment. One of the main strengths of the SBM is that it incorpo- rates variables which may include both subjective (e.g. health beliefs, perception of illness) and objective (e.g. income, symptoms) variables from a variety of domains – socioeconomic, biological, psychological and social. http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 other. However, it is not specifically related to health and does not include factors relating to emotional and interac- tive aspects of care which may be important for both CAM use and child healthcare. The Consumer Decision Making model contained no integration of affective or value- laden factors which would differentiate individuals with similar experience from one another[44]. importance of health beliefs and organisational (ena- bling) factors may be underestimated due to the inade- quate conceptualisation (and therefore measurement) of these in many studies. Findings have supported Andersen's claim that need factors are important, but it should be emphasised that these factors are heavily dependent on social context and health beliefs[50]. CAM definition 2 & $ 2 # 2 4 * " 0 2 U # $ ' ! " M N O P Q R Factors in the SBM important for CAM use Figure 2 Factors in the SBM important for CAM use. References show which factors were important in which studies. Page 7 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 Authors' contributions MB conceived of the paper. NR supervised the project. YIC carried out the literature searches and YIC and AL reviewed individual papers. The manuscript was drafted by AL and YIC and revised and amended by MB and NR. All authors read and approved the final manuscript. Health behaviour models The Health Locus of Control, Self-Regulatory model, TPB and TTM all had the weakness that they tended to origi- nate from a singular viewpoint which results in limited integration of the sources of influence, and thus they were The Consumer Decision-Making model[51] was able to take into account the variability in the decision factors and the intricacy of their relationships and effects on each Page 8 of 12 (page number not for citation purposes) Page 8 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 able to account for limited variability in the dependent variable. studies are often omitted from subject analysis as they do not fit easily into pre-set conceptual categories. As child use of CAM was identified as an underexplored area, the use of qualitative methodology to examine the predictors and correlates of CAM use would be particularly relevant for child use of CAM. The models developed using qualitative data may prove useful once empirically tested, although these were all based on patients with chronic health prob- lems[71,75,77]. Some of the authors are now engaged in a funded research project using qualitative methods to clarify validity and to identify the relative importance of the factors and will test this using a quantitative questionnaire using correlational and regression analysis to validate the model. Abbreviations CAM: Complementary and alternative medicine; CM: Conventional medicine; SBM: Sociobehavioural model; HLoC: Health Locus of Control; TTM: Transtheoretical model; TPB: Theory of planned behaviour; SRM: Self reg- ulatory model. Future research The review found that the SBM has strong support for modelling the decision making process in CAM use. How- ever, a number of methodological limitations were iden- tified which future research needs to address. The decision making process appears to vary depending on the CAM modality; comparison between CAM modalities should be made. Extraneous variables should be controlled for, especially illness characteristics. Quantitative studies should include sufficiently powered samples, validated measures and multivariate analysis. Studies of the SBM should also incorporate the dynamic, interactive nature of the factors in the model. Additional material The use of qualitative methods to explore decision mak- ing is particularly interesting and should be considered carefully. The discipline of psychology is particularly prone to quantitative methods, except when the subject of study is exploratory. This issue may represent a disadvan- tage in applying psychological approaches to the data as many pertinent findings which arise from qualitative Additional file 1 Results of literature review. This table gives detailed information on each study included in the review, including study design, factors predict- ing CAM use, limitations and suitability of the model tested. Click here for file [http://www.biomedcentral.com/content/supplementary/1472- 6882-9-9-S1.doc] Additional file 1 Results of literature review. This table gives detailed information on each study included in the review, including study design, factors predict- ing CAM use, limitations and suitability of the model tested. Click here for file [http://www.biomedcentral.com/content/supplementary/1472- 6882-9-9-S1.doc] Limitations of review Due to disparate terms used in the decision making liter- ature, it was difficult to define search terms to capture all relevant papers. Although searches were kept broad with extensive hand searching of reference lists to capture a wide range of articles, the search terms could have included terms such as 'Framework', although subsequent scanning of results it did not seem to make a difference to the papers chosen for inclusion. Language bias may well be an issue as only English language papers were chosen (15 were excluded for this reason) [90]. p g The authors declare that they have no competing interests. p g The authors declare that they have no competing interests. Conclusion d Andersen's sociobehavioural model has been identified as a suitable model for modelling the decision making proc- ess resulting in adult CAM use. However, the suitability of application of this model to child CAM use has not fully been studied and needs further clarification. This identifi- cation of a suitable decision making model is facilitating theory-guided research into how and why CAM is used for children, through empirical testing. Using an existing model promotes methodological consistency, which is imperative in the field of CAM which often uses disparate methods and tools. Providing an overarching model which has been tested for a child population will aid to guide healthcare practitioners' understanding and appli- cation to clinical practice[91]. The review only included published papers, which did not capture potentially important sources of information such as theses, conferences abstracts and official reports. In addition papers on this subject were published in journals in a very wide range of subject areas; there may be other databases which should have been included. The review only included studies published post 1994 which may have been a source of bias. Terms for CAM could have been expanded to include all CAM modalities (e.g. acupuncture, herbal etc), in order to capture studies that may not be indexed under general CAM terms. Health behaviour models As such this model fits very comfortably with an interdisci- plinary and integrative view of healthcare utilisation, whist taking account of idiosyncratic influences as well. TTM and TPB were found to be supportive of CAM use prediction and psychological factors were more important than medical or demographic, but only when the two models were used together, and findings may be open to selection bias[63]. Both beliefs about the positive effects and worries about the negative effects were impor- tant[63]. Family expectation was particularly important in the TPB[63]. However, the SBM falls short of naming the specific proc- esses which are often complicated and non-linear, which lead to the specific decisions to use CAM among individ- uals and subgroups[44]. Also, qualitative studies [73,77] highlighted the importance of temporal factors, e.g. decid- ing to use CAM through continuous appraisal of well- being and due to perception of circumstances at that point, which SBM fails to account for. While the model succeeds in incorporating health beliefs within the predis- posing factors, which in turn encompass other affective and cognitive factors, specific health beliefs are often not identified. Empirical study using the SBM needs to ensure that the findings are integrated back into the model and not left as a collection of associations. To this end, it is rec- ommended that future studies utilise a longitudinal pro- spective approach, whilst ensuring differentiation between different CAM modalities. The self-regulatory model received little support for pre- dicting CAM use[70]. People may pursue a particular treatment if they perceive their illness in a certain way or hold particular treatment beliefs, for example having a holistic approach to health and illness being the strongest predictor of CAM use[89]. Braden's self help model had strong support as patients used CAM because it was per- ceived as effective and was related to income, however it was only tested by one study of cancer patients[72]. References 1. Defining and describing complementary and alternative medicine. Panel on Definition and Description, CAM Research Methodology Conference, April 1995. Altern Ther Health Med 1997, 3:49-57. 25. Kleinman A: Patients and Healers in the Context of Culture: An exploration of the Borderland between Anthropology, Medicine and Psychiatry Berkeley and Los Angeles: University of California Press; 1981. 2. Ernst E, Resch KL, Mills S, Hill R, Mitchell A, Willoughby M, White A: Complementary Medicine – a definition. Br J Gen Pract 1995, 45:506. 26. Patterson C, Arthur H, Noesgaard C, Caldwell P, Vohra J, Francoeur C, Swinton M: Exploring adolescent complementary/alterna- tive medicine (CAM) use in Canada. Journal of Interprofessional Care 2008, 22:45-55. 3. Zollman C, Vickers A: ABC of complementary medicine: What is complementary medicine? British Medical Journal 1999, 319:693-696. 27. 27. Prussing E, Sobo EJ, Walker E, Kurtin PS: Between 'desperation' and disability rights: a narrative analysis of complementary/ alternative medicine use by parents for children with Down syndrome. Social Science & Medicine 2005, 60:587-598. 4. Pachter LM, Sumner T, Fontan A, Sneed M, Bernstein BA: Home- based therapies for the common cold among European American and ethnic minority families: The interface between alternative/complementary and folk medicine. Archives of Pediatrics & Adolescent Medicine 1998, 152:1083-1088. 28. 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Additional file 1 Page 9 of 12 (page number not for citation purposes) Page 9 of 12 (page number not for citation purposes) http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 http://www.biomedcentral.com/1472-6882/9/9 BMC Complementary and Alternative Medicine 2009, 9:9 Acknowledgements 23. Coyne I: Children's participation in consultations and deci- sion-making at health service level: A review of the litera- ture. International Journal of Nursing Studies 2008, 45:1682-1689. g We would like to thank Rizvana Ahmed and Julie Donaldson and the King's Fund Partners for Health in London programme for the project grant. 24. Rhodes P, Small N, Ismail H, Wright J: The use of biomedicine, complementary and alternative medicine, and ethnomedi- cine for the treatment of epilepsy among people of South Asian origin in the UK. BMC Complementary and Alternative Medi- cine 2008, 8:7. 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Leventhal H, Diefencach M, Leventhal EA: llness cognition: using common sense to understand treatment adherence and affect cognition internactions. Cognitive Therapy and Research 1992, 16:143-163. Pre-publication history The pre-publication history for this paper can be accessed here: http://www.biomedcentral.com/1472-6882/9/9/prepub http://www.biomedcentral.com/1472-6882/9/9/prepub http://www.biomedcentral.com/1472-6882/9/9/prepub Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." 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5deef6edaceb406e782848d88bd9fe6c_1	French-Science-Pile	Open Science	Various open science	2,015	High frequency waves and the maximal smoothing effect for nonlinear scalar conservation laws. SIAM Journal on Mathematical Analysis, 2014, 46 (3), pp.2160-2184. ⟨10.1137/120880367⟩. ⟨hal-00576662v3⟩	None	Unknown	Unknown	8,031	13,264	High frequency waves and the maximal smoothing effect for nonlinear scalar conservation laws Stéphane Junca To cite this version: Stéphane Junca. High frequency waves and the maximal smoothing effect for nonlinear scalar conservation laws. SIAM Journal on Mathematical Analysis, 2014, 46 (3), pp.2160-2184. �10.1137/120880367�. �hal-00576662v3� HAL Id: hal-00576662 https://hal.science/hal-00576662v3 Submitted on 21 Jun 2015 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. HIGH FREQUENCY WAVES AND THE MAXIMAL SMOOTHING EFFECT FOR NONLINEAR SCALAR CONSERVATION LAWS STÉPHANE JUNCA Abstract. The article first studies the propagation of well prepared high frequency waves with small amplitude ε near constant solutions for entropy solutions of multidimensional nonlinear scalar conservation laws. Second, such oscillating solutions are used to highlight a conjecture of Lions, Perthame, Tadmor, ([23]), about the maximal regularizing effect for nonlinear conservation laws. For this purpose, a definition of smooth nonlinear flux is stated and compared to classical definitions. Then it is proved that the uniform smoothness expected by [23] in Sobolev spaces cannot be exceeded for all smooth nonlinear fluxes. Key-words: multidimensional conservation laws, nonlinear smooth flux, geometric optics, Sobolev spaces, smoothing effect. Mathematics Subject Classification: Primary: 35L65, 35B65; Secondary: 35B10, 35B40, 35C20. Contents 1. Introduction 2. The smoothing effect in Sobolev spaces 3. Main Results 3.1. Critical geometric optics 3.2. Consequence on the uniform maximal smoothing effect 4. High frequency waves with small amplitude 5. Characterization of nonlinear flux 5.1. Nonlinear smooth flux 5.2. Comparisons with other nonlinear flux definitions 6. Sobolev estimates 7. Super critical geometric optics and maximal smoothing effect References 1 3 5 6 7 10 12 13 17 19 28 29 1. Introduction This paper deals with super critical geometric optics to highlight the maximal regularizing effect for nonlinear multidimensional scalar conservation laws. Date: April 8, 2014. 1 2 STÉPHANE JUNCA This effect is studied in Sobolev spaces by P.L. Lions, B. Perthame and E. Tadmor in [23]. They obtain a uniform fractional Sobolev bounds for any ball of L∞ initial data under a non linearity condition on the flux quantified by a parameter α ∈]0, 1]. They also conjectured the better Sobolev exponent s = α for entropy solutions. For the first time, the multidimensional case is investigated to bound this maximal smoothing effect. Furthermore, all smooth nonlinear fluxes are considered in this paper. We build sequences of solutions uniformly bounded s,1 in Wloc with the conjectured maximal exponent s = α and with no possible improvement of the Sobolev exponent. High frequency periodic solutions are used for this purpose. Near a constant state and for L∞ data, geometric optics expansions with various frequencies and various phases are validated in the framework of weak entropy solutions and of L1loc convergence in [5]. Here, results of [5] are specified in C 1 for a well chosen phase and proved for a particular sequence of smooth solutions (without shocks on a strip). This allows us to prove that, necessarily, the best uniform Soblev exponent s, for entropy solutions and for positive time, satisfies s ≤ α for a ball of L∞ initial data. Notice that we look for the best uniform Sobolev exponent for a set of solutions. The smoothness of any individual solution is not studied in this paper. This point is discussed later. An important point to note here is the definition of nonlinear flux. In [23] they give well known Definition 2.4 below and the conjecture about the maximal smoothing effect in Sobolev spaces related to the parameter “α“ from their definition. The study of periodic solutions leads to another definitions [15, 5]. We obtain Definition 5.1 for smooth flux. It generalizes the definition of [5] and it is typical in the context of stationary phase. For C ∞ flux, our definition is equivalent to classical Definition 2.4. Furthermore, Definition 5.1 gives a way to compute the parameter “α”. Our definition also shows that smoothing effects for scalar conservation laws depend on the space dimension. The paper is organized as follows. To be more precise, the smoothing effect and the related conjecture in the Sobolev framework are recalled in Section 2. Section 3 contains the two main results, propagations of high frequency waves and the consequence for the maximal smoothing effect. Some comments and other approaches are also discussed. In Section 4 examples of highly oscillating solutions are validated under new orthogonality conditions between the flux derivatives and the phase gradient. In Section 5, these orthogonality conditions lead to a new definition of nonlinear smooth flux. The concept of flux non-linearity is clarified , characterized and compared with other classical definitions. Section 6 is devoted to get optimal Sobolev estimates on oscillating solutions built in Section 4. It is a quite technical part. Finally, Section 7 gives the super critical geometric optics expansion with the highest frequency related to the geometric structure of the nonlinear flux. The existence of this family of high frequency waves implies s ≤ α. But the whole conjecture s = α is still an open problem. HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 3 2. The smoothing effect in Sobolev spaces We look for Sobolev bounds for entropy solutions u(., .) of (2.1) ∂t u + divx F(u) = 0, where t ∈ [0, +∞[, x ∈ Rd , u : [0, +∞[t ×Rdx → R, F : R → Rd is a smooth flux function, F ∈ C ∞ (R, Rd ), and the initial data are only bounded in L∞ (Rdx , R): (2.2) u(0, x) = u0 (x). The smoothing effect depends on the class of solutions studied. It is well known that shock occurs even with smooth initial data [10]. Let us recall definitions of solutions for initial-value problem (2.1),(2.2). Definition 2.1. [Weak solutions] We say that a function u ∈ L∞ ([0, +∞[×Rd ) is a weak solution of initial-value problem (2.1),(2.2) provided Z Z +∞ Z u0 (x) φ(0, x)dx = 0 u ∂t φ + F(u) · ∇x φ dx dt + 0 Rd Rd for all smooth functions φ with compact support. This definition means that partial differential equation (2.1) is written in the sense of distribution. This class of solutions is too large to provide uniqueness and smoothing effect. We have to restrict our attention to a more physical class of solutions. Definition 2.2. [entropy solutions] We say that a function u ∈ L∞ ([0, +∞[×Rd ) is an entropy solution of initialvalue problem (2.1),(2.2) provided the following two conditions hold: • ∂t ϕ(u) + divx Q(u) ≤ 0 in the sense of distribution, for all convex functions ϕ where Q0 = ϕ0 F0 , ϕ is called an entropy and Q its entropy-flux, • u ∈ C 0 ([0, +∞[, L1loc (Rd , R) and lim u(t, x) = u0 (x) in L1loc (Rd , R). t→0 It is classical to check that an entropy solution is a weak solution. It suffices to take ϕ(u) = ±u and to recover the weak trace u0 at t = 0 since the second condition means that u has a strong trace at t = 0. For this class of solution, Kruzkov proved in 70’ the uniqueness (see for instance [10] and the references therein) and Lions, Perthame, Tadmor proved a smoothing effect in Sobolev spaces [23]. Indeed, the smoothing effect and the proof given in [23] is valid in the following larger class of solutions. Definition 2.3. [Solutions with bounded entropy production] We say that a weak solution of initial-value problem (2.1),(2.2) has a bounded entropy production provided for each entropy ϕ and entropy-flux Q, there exists a signed measure µ, locally bounded, such that ∂t ϕ(u) + divx Q(u) = µ. 4 STÉPHANE JUNCA The uniqueness is lost for this class, but, since the fundamental paper [23], various results and interests can be found in [11, 13, 16]. Indeed, for entropy solutions, the measure is non positive, µ ≤ 0 and for smooth solutions, µ ≡ 0. The smoothing effect for the class of solutions with bounded entropy production is clearly weaker than the smoothing effect for the class of entropy solutions since the first class is larger than the second class. Let us turn to the notion of nonlinear flux. Set a(u) = F0 (u). Obviously, when F is linear: a(u) = a where a is a constant vector, u(t, x) = u0 (x − t a) so that there is no smoothing effect. In [23], it was first proved a regularizing effect for nonlinear multidimensional flux F. The sharp measurement of the non-linearity plays a key role in our study. Let us recall the classical definition for nonlinear flux from [23]. Definition 2.4. [ Nonlinear flux [23]] Let M be a positive constant. F : R → Rd is said to be nonlinear on [−M, M ] if there exist α > 0 and C = Cα > 0 such that for all δ > 0 (2.3) supτ 2 +\|ξ\|2 =1 \|Wδ (τ, ξ)\| ≤ C δ α , where (τ, ξ) ∈ S d ⊂ Rd+1 , i.e. τ 2 + \|ξ\|2 = 1, and \|Wδ (τ, ξ)\| is the one dimensional measure of the singular set: Wδ (τ, ξ) := {\|v\| ≤ M, \|τ + a(v) ξ\| ≤ δ} ⊂ [−M, M ] and a = F0. Indeed, Wδ (τ, ξ) is a neighborhood of the cricital value v for the symbol of the linear operator L[v] in the Fourier direction (τ, ξ) where L[v] = ∂t + a(v) ∇x. The symbol in this direction is: i (τ + a(v) ξ). This operator is simply related to any smooth solution u of equation (2.1) by the chain rule formula: ∂t u + divx F(u) = ∂t u + a(u) ∇x u = L[u]u. α is a degeneracy measurement of the operator L parametrized by v. α depends only on the flux F and the compact set [−M, M ]: α = α[F, M ]. In the sequel we denote by (2.4) αsup = αsup [F, M ], the supremum of all α satisfying (2.3). α, or more precisely αsup , is a key parameter to describe the sharp smoothing effect for entropy solutions of nonlinear scalar conservation laws. For smooth flux the parameter α always belongs to [0, 1], for instance: αsup = 0 for a linear flux, α = 1 for strictly convex flux in dimension one. For the first time αsup is characterized below in Section 5. Indeed, for smooth nonlinear flux, 1 is always an integer greater or equal to the space dimension. αsup In the sequel we assume that M ≥ ku0 k∞ and the flux F is nonlinear on [−M, M ] so that: (2.5) αsup > 0. HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 5 When nonlinear condition (2.5) is true, the entropy solution operator associated with the nonlinear conservation law (2.1), (2.2), St : L∞ (Rdx , [−M, M ]) → L∞ (Rdx , [−M, M ]) u0 (.) 7→ u(t, .), s,1 (Rdx , R) for all has a regularizing effect: mapping L∞ (Rdx , [−M, M ]) into Wloc t > 0. α In [23], they proved this regularizing effect for all s <. 2+α α under a generic assumption In [31] the result is improved for all s < 1 + 2α 0 00 on a = F . These results are based on averaging lemmas and are still valid for solutions with bounded entropy production (Definition 2.3). For entropy solutions, Lions, Perthame and Tadmor conjectured in 1994 a better regularizing effect, ([23], remark 3, p .180, line 14-17). In [23], they proposed an optimal bound ssup for Sobolev exponents of entropy solutions: (2.6) ssup = αsup. s,1 That is to say, u(t, .) belongs in all Wloc (Rd , R) for all s < ssup = αsup and for all t > 0. The shock formation implies s < 1 and ssup ≤ 1 since W 1,1 functions do not have shock. The main goal of the paper is to give an insight of the conjecture (2.6) by bounding the uniform Sobolev smoothing effect ssup for the whole set of entropy solutions with initial data bounded by M in L∞ : (2.7) ssup ≤ αsup. Some previous results highlight the conjecture (2.6) or the inequality (2.7) in the one dimensional case. This is discussed in Subsection 3.2.1 for entropy solutions and for solutions with bounded entropy production. But for the multidimensional case and for all smooth fluxes, our results are new. 3. Main Results Our main results are about the propagation of high frequency waves with frequencies higher than usual [14, 29] and the consequence on the uniform maximal smoothing effect for scalar conservation laws. Weakly nonlinear geometric optics study propagations of sequences of high frequency waves. The classical case [14, 29] deals with uniformly bounded derivatives. That is to say the sequence is bounded in W 1,1 or in BV when shocks occur. We show that sequence of high frequencies waves, not uniformly bounded in W 1,1 , can propagate under a stationary phase assumption on the flux, condition (3.5) below. Since the frequencies are higher than usual in geometric optics, we call this case critical geometric optics. A natural question is about the maximal frequencies arising in critical geometric optics. This last case is called super critical geometric optics. Critical geometric optics is expounded in Subsection 3.1. The explicit construction of the super critical optics are expounded later and completed in the 6 STÉPHANE JUNCA last section, Section 7. A main consequence of super critical geometric optics on smoothing effect is stated and commented in Subsection 3.2. 3.1. Critical geometric optics. This subsection deals with highly oscillating initial data near a constant state: v · x (3.1) uε (0, x) = uε0 (x) := u + εU0 , εγ where U0 (θ) is a one periodic function w.r.t. θ, γ > 0, u is a constant ground state, u ∈ [−M, M ], v ∈ Rd . The case γ = 1 is the classical geometric optics for scalar conservation laws, ([14]). In this paper we focus on critical oscillations, that is to say γ > 1. The aim of this section is to see when such high frequency are propagated or not propagated. As we will see, it depends on new compatibility conditions between the phase and the flux (3.5). One of the two following asymptotic expansions (3.2) or (3.3), is expected in 1 Lloc (]0, +∞[×Rd , R) for the entropy-solution uε of conservation law (2.1) with highly oscillating data (3.1) when ε goes to 0, φ(t, x) uε (t, x) = u + εU t, (3.2) + o(ε) εγ (3.3) or uε (t, x) = u + εU 0 + o(ε), whereZthe profile U (t, θ) satisfies a conservation law with initial data U0 (θ), 1 U0 (θ)dθ and the phase φ satisfies the eikonal equation: U0 = 0 (3.4) ∂t φ + a(u) ∇x φ = 0, φ(0, x) = v · x. Thus the phase is simply a linear phase: φ(t, x) = v · (x − t a(u)). The propagation of such oscillating data is obtained under the crucial compatibility condition (3.5) below. On the other hand, when the the compatibility condition (3.5) is nowhere satisfied, the nonlinear semi-group associated with equation (2.1) cancels these high oscillations, see Theorem 4.1. The validity or invalidity of assumption (3.5) is a key point related to the nonlinearity of the flux (Section 5). Theorem 3.1. [Propagation of smooth high oscillations] Let γ belong to ]1, +∞[ and let q be the integer such that q − 1 < γ ≤ q. Assume F belongs to C q+3 (R, Rd ), U0 ∈ C 1 (R/Z, R), v 6= (0, · · · , 0) and (3.5) a(k) (u) v = 0, k = 1, · · · , q − 1. Then there exists T0 > 0 such that, for all ε ∈]0, 1], the solutions of conservation law (2.1) with initial oscillating data (3.1) are smooth on [0, T0 ] × R and φ(t, x) uε (t, x) = u + εU t, + O(ε1+r ) in C 1 ([0, T0 ] × Rd ), εγ HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 7 1 if γ = q, q − γ else, φ is given by the eikonal equation (3.4) and the smooth profile U is uniquely determined by the Cauchy problem (3.6): where 0 < r = (3.6) with b = 1 (q+1)! ∂U q+1 ∂U +b = 0, U (0, θ) = U0 (θ), ∂t ∂θ a(q) (u) v if γ = q,. 0 else. We deal with smooth solutions to compute Sobolev bounds later. Indeed, the asymptotic stays valid after shocks formation and for all positive time but in L1loc instead of L∞ ([5]). When γ = 1, we do not need assumption (3.5). It is the classic case for geometric optics ([14, 29]). In dimension d ≥ 2, it is always possible to find a non trivial vector v satisfying (3.5). For example for γ = 2, (3.5) is reduced to find v 6= 0 such that a0 (u) v = 0. Thus, such singular solutions always exist in dimension greater than one. But, for genuine nonlinear one dimensional conservation law, there is no such solution. Of course, we assume that U0 is a non constant function and that F is a nonlinear function near u, else the theorem is obvious. Indeed, when U0 is constant, uε is also constant. When F is linear on [u − δ, u + δ] for some δ > 0, high oscillations propagate for all time without any restriction of the phase and of the frequency size. In fact, Theorem 3.1 expresses a kind of degeneracy of multidimensional scalar conservation laws. This degeneracy (period smaller than the amplitude) appears for quasilinear systems whit some nonlinear degeneracy (see for instance [7]). Notice that for γ > 1, smooth solutions exist for larger time than it is currently known [10, 22]: Tε ∼ 1/\|∇x uε0 \| ∼ εγ−1 . Furthermore, equation (3.6) is nonlinear if and only if γ ∈ N and aq (u) v 6= 0. 3.2. Consequence on the uniform maximal smoothing effect. We construct a sequence of smooth solutions which are exactly uniformly bounded in the Sobolev space conjectured in [23]. The uniform Sobolev estimate of this sequence blows up in all more regular Sobolev spaces. This super critical geometric optics expansion and its optimal Sobolev estimates are the goal of this paper which is completed in Section 7. Let us state the main consequence of this construction in the following theorem. Theorem 3.2. [Bound of the maximal smoothing effect] Let F be a nonlinear flux which belongs to C ∞ ([−M, M ], Rd ). Let αsup be the sharp measurement of the flux non-linearity. Then there exist a constant u ∈ [−M, M ], a time T0 > 0, and a sequence of initial data (uε0 )0<ε<1 such that 8 STÉPHANE JUNCA kuε0 − ukL∞ (Rd ) < ε, and the sequence of entropy solutions (uε )0<ε<1 associated with conservation law (2.1) satisfy the followings: • for all s ≤ αsup , the sequence (uε )0<ε<1 is uniformly bounded s,1 s,1 (Rd )), ([0, T0 ] × Rd ) ∩ C 0 ([0, T0 ], Wloc in Wloc • for all s > αsup , the sequence (uε )0<ε<1 is unbounded s,1 s,1 in Wloc ([0, T0 ] × Rd ) and in C 0 ([0, T0 ], Wloc (Rd )). Theorem 3.2 gives the upper bound (2.7) for the W s,1 −regularizing effect : ssup ≤ αsup . Indeed, let us denote St the semi-group associated with conservation law (2.1) and B ∞ (u, ρ)) = {u ∈ L∞ (Rd , R), ku − ukL∞ (Rd ,R) < ρ}. Theorem 3.2 proves that for a well chosen u ∈ [−M, M ], there exists T0 > 0, such that for all ρ > 0 and for all 0 < t < T0 , St (B ∞ (u, ρ)) is not a bounded s,1 subset of Wloc (Rdx ) for all s > αsup . This result yields several remarks. Remark 3.1. Optimality for large dimension or small nonlinear degeneracy. 1 By Theorem 5.1, αsup ≤ , so, for larger dimension, αsup is smaller. Using d a better lower bound of ssup from [31] we have: αsup 1 ≤ ssup ≤ αsup ≤. 1 + 2 αsup d Thus for large dimension (d >> 1) or small nonlinear degeneracy (αsup << 1) we have asymptotically: ssup ∼ αsup. Remark 3.2. In W s,p , 1 < p < +∞, our geometric optics expansion shows that spsup ≤ αsup by Theorem 6.1, where spsup denotes the maximal uniform smoothing effect in W s,p . In other words our example is not related to the parameter p. Indeed, our geometric optics expansion is bounded in W s,∞. Other examples show the importance of the parameter p in Subsection 3.2.1. Sobolev spaces are not sufficient to describe all the properties of the solutions. Some comments are given in Subsection 3.2.1 for other approaches. 3.2.1. Other approaches for the smoothing effect. The maximal Sobolev exponent is not sufficient to characterize the smoothing effect. Other relevant ways are presented by one-sided Oleinik condition, BV , generalized characteristics, other oscillating solutions, BV s , trace properties. First, we comment other approaches on the one dimensional case, where the optimal smoothing effect is not yet completely understood . Second, we briefly discuss the multidimensional case where there are only few results. One dimensional case: • α = 1, s = 1, entropy solutions. In one dimension (d=1) and for uniformly convex flux, it is well known from Lax and Oleinik that the entropy solution becomes BV , HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 9 ([24, 22]). Conjecture (2.6) is true in this case since for all t > 0, u(t, .) s,1 for all s < 1. belongs to Wloc • α = 1, s = 1/3: for solutions with bounded entropy production. For Burgers flux, F (u) = u2 , De Lellis and Westdickenberg built piecewise constant solutions with bounded entropy production to show that s ≤ 1/3 [13]. Recently, for uniform convex flux (α = 1), the optimal Sobolev exponent s = 1/3 is reached in [16]. • 0 < α < 1, s = α: entropy solutions. For power law flux, F (u) = \|u\|1+p ,p ≥ 1, De Lellis and Westdickenberg also built piecewise smooth entropy solutions and proved that s ≤ α ([13], Proposition 3.4 p. 1085). For all nonlinear smooth fluxes, new continuous examples are also given in [4]. Both examples are only justified for a bounded time interval before waves interact. For nonlinear degenerate convex fluxes the regularity ssup = αsup is reached in W s,1 ([19]) and also W s,1/s ([3]). For this purpose, fractional BV spaces, which are called BV s , are introduced in [3]. BV s functions have a structure similar to the one of BV maps, for all s ∈]0, 1]. BV s spaces seem to be the natural spaces to capture the optimal regularizing effect for one dimensional scalar conservation laws. • One-sided Oleinik condition and its generalizations: In the 50’, Oleinik ([24]) obtained her one-sided Lipschitz condition which ensures the uniqueness and the BV regularity of the entropy solution. This is the first basis and the proof of s = α for one dimensional uniformly convex flux (α = 1). Dafermos ([9, 10]), with his generalized characteristics, handled convex and some degenerate convex fluxes. Hoff extended this one-sided condition in several space variables ([17]) but restricted to vectorial fluxes which are scalar convex fluxes after a change of space variables. A generalized Oleinik condition, only validated in the one dimensional case, is the key assumption to prove the best W s,1 smoothing effect in [19] (0 < s = α < 1). The maximal W s,p smoothing effect is proved in [3] with a one-sided Hölder condition and BV s spaces. For a recent generalization of Oleinik condition for a flux with one inflection point, we refer the reader to [20]. To conclude, the one-sided Oleinik condition is essentially specific to the one dimensional case with some convexity on the flux. Multidimensional case: In the 90’, the kinetic formulation of conservation laws ([23]) gave another approach for other authors. This approach is also valid for solutions with bounded entropy production. In this framework, some suitable averaging of nonlinear expressions of the solution are BV ([6]). Some trace properties were 10 STÉPHANE JUNCA first obtained in [34, 11, 12, 8]. These structure of a BV function for solutions, without being BV , cannot be given by Sobolev regularity. Neither BV nor W s,p is the perfect space to explain properties of solutions with only L∞ initial data. 4. High frequency waves with small amplitude In this section we prove Theorem 3.1 about the propagation of oscillations with not uniformly bounded derivatives, that is to say space or time derivatives unbounded with respect to the parameter ε. We also show the optimality of the assumption (3.5) in Theorem 4.1. Proof of Theorem 3.1: First one performs a WKB computations with following ansatz: φ(t, x) uε (t, x) = u + ε Uε t, (4.1). εγ Notice that we use the exact profile Uε for the proof as in [21]. It is a method to sharply control the difference between the exact solution and the geometric optics expansion: Uε and U. The Taylor expansion of the flux and the remainder are: q+1 X Uεk (k) F (u) + εq+2 Gεq (Uε ), k! k=0 Z 1 (1 − s)q+1 (q+2) ε q+2 F (u + sεU )ds, Gq (U ) = U (q + 1)! 0 gqε (U ) = v.Gεq (U ). F(uε ) = εk We now compute the partial derivatives with respect to time and space variables: φ(t, x) ∂t Uε t, = ∂t Uε − ε−γ (a(u) · v)∂θ Uε γ ε divx F(uε ) = q X εk+1−γ k=0 1−γ = ε where cq = ∂θ Uεk+1 (k) a (u) · v + εq+2 divx Gεq (Uε ) (k + 1)! (a(u) · v)∂θ Uε + εq+1−γ cq ∂θ Uεq+1 + εq+2−γ ∂θ gqε (Uε ), a(q) (u) · v . Then simplification yields (q + 1)! (4.2) ∂t uε + divx F(uε ) = ε ∂t Uε + εq−γ cq ∂θ Uεq+1 + ε1+q−γ ∂θ gqε (Uε ). It suffices to take Uε to be the solution of the one dimensional scalar conservation laws with ψε (U ) = εq−γ cq U q+1 + ε1+q−γ gqε (U ) (4.3) ∂t Uε + ∂θ ψε (Uε ) = 0, Uε (0, θ) = U0 (θ). 2 2 Notice that ψε = O(1) ∈ Cloc . For γ < q, ψε is even smaller: ψε = O(εr ) ∈ Cloc. That is enough to prove the existence of a sequence of smooth oscillating HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 11 solutions on the same strip. Uniform life span for smooth solutions (Uε )0<ε≤1 : d We use the method of characteristics with ψε0 (U ) = dU ψε (U ): d Θ(t, θ) = ψε0 (Uε (t, Θ(t, θ)), Θ(0, θ) = θ. dt Since Uε is constant along the characteristics, Θ(t, θ) = θ + tψε0 (U0 (θ)). As long as the map θ → Θ(t, θ) is not decreasing no shock occurs. ∂ d Θ(t, θ) = 1 + tψε00 (U0 (θ)) U0 (θ) ∂θ dθ The first shock appears at the time Tε when the right hand side vanishes. Let d m0 = sup[0,1] \|U0 \| > 0, d0 = sup[0,1] U0 , m = sup0<ε≤1 sup\|U −u\|≤m0 \|ψε00 (U )\|, dθ then, d 00 1/Tε = sup[0,1] −ψε (U0 (θ)) U0 (θ) ≤ m d0. dθ Of course, for constant initial data (d0 = 0), no shock occurs, the solution is constant and Tε = +∞. In the case m d0 6= 0, Tε is finite but 0 < inf0<ε≤1 Tε since Tε ≥ 1/(m d0 ) for all 0 < ε ≤ 1. This gives the existence of a positive time T0 < T ∗ = inf{Tε , ε ∈]0, 1]} such that Uε ∈ C 1 ([0, T0 ] × R/Z). Thus uε , which is well defined by (4.1), belongs to C 1 ([0, T0 ] × Rd ) for all 0 < ε ≤ 1. Now we prove the C 1 convergence of the geometric optics expansion. There are two cases: γ is an integer or not. q = γ: From (4.2) and (2.1) we get ∂t Uε + ∂θ cq Uεq+1 + εgqε (Uε ) = 0, ∂t U + cq ∂θ U q+1 = 0, Uε (0, θ) = U0 (θ), U (0, θ) = U0 (θ). The method of characteristics gives C 1 characteristics, C 1 solutions and kUε − U kC 1 ([0,T0 ]×Rd ) = O(ε), where kU kC 1 ([0,T0 ]×Rd ) = kU kL∞ ([0,T0 ]×Rd ) + k∂t U kL∞ ([0,T0 ]×Rd ) + k∂θ U kL∞ ([0,T0 ]×Rd ). integer q > γ: The proof is similar except that the term εr cq ∂θ (cq U q+1 ) becomes a remainder, with r = q − γ and U (t, θ) = U0 (θ), thus kUε (., .) − U0 (.)kC 1 ([0,T0 ]×Rd ) = O(εr ), which concludes the proof. When condition (3.5) is violated, oscillations are immediately canceled. 12 STÉPHANE JUNCA Theorem 4.1. [Cancellation of high oscillations, [5]] Let F belong to C q+2 and U0 ∈ L∞ (R/Z, R), where q − 1 < γ ≤ q where q is defined in Theorem 3.1. If for some 0 < j < q (4.4) a(j) (u) v 6= 0 then the solutions uε of conservation law (2.1) with initial oscillating data (3.1) for ε ∈]0, 1] satisfy when ε → 0 uε (t, x) = u + εU 0 + o(ε) in L1loc (]0, +∞[×Rd ). Obviously the interesting case is when U0 is non constant. When U0 is smooth and non constant the first time when a shock occurs Tε converges towards 0 when ε → 0. Thus solutions are weak entropy solutions. The proof is in the spirit of [5] and uses averaging lemmas (see [27] and the references given there). The proof is briefly expounded to be self-contained. Proof : For non constant initial data it is impossible to avoid shock waves on any fixed strip [0, T0 ] × Rd with T0 > 0 as in the previous proof of Theorem 3.1 since the time span of smooth solutions is εβ where β = γ − j > 0. First, with a change of space variable x ↔ x − t.a(u), we can assume that a(u) = 0. The WKB computations use the following anzatz: uε (t, x) = u + εvε (t, x) where vε (t, x) = Wε (t, ε−j φ(t, x)). Indeed, the condition (4.4) leads to such anzatz as we can see in the WKB computations of the proof of Theorem 3.1. Then Wε satisfies the one dimensional nonlinear conservation laws: (4.5) ∂t Wε + ∂θ cj Wεj+1 + εgjε (Wε ) = 0, Wε (0, θ) = U0 (ε−β θ), cj 6= 0. Wε (0, .) converges weakly towards U 0 . As in [5], Wε is relatively compact in L1loc thanks to averaging lemmas. Then Wε converges towards the unique entropy solution W of ∂t W + cj ∂θ W j+1 = 0, W (0, θ) = U 0. That is to say that W (t, θ) ≡ U 0 . Then vε (t, x) converges towards U 0 in L1loc which completes the proof. 5. Characterization of nonlinear flux The flux nonlinearity is characterized by the parameter α in Definition 2.4, the Lions-Perthame-Tadmor definition of nonlinear flux. The smoothing effect depends only on the best α = αsup . The understanding of the parameter αsup is a key step to the comprehension of the regularity of entropy solutions. Unfortunately, there are only few examples where αsup is computed in dimension 2 ([23, 31]) and there are some remarks in [18, 19, 2]. For the first time, for all nonlinear C ∞ fluxes and for all dimensions we characterize the fundamental parameter αsup . For this purpose we state Definition 5.1 of smooth nonlinear flux. This definition is related to the critical HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 13 geometric optics expansion given in Theorem 3.1. Notice that Definition 5.1 is typical for stationary phase methods. It is less common in the context of conservation laws. Let us emphasize on two important consequences of Definition 5.1. • The parameter αsup is explicitly characterized with the flux derivatives in Theorem 5.1. • The super critical geometric optics expansion is built to highlight the uniform maximal smoothing effect in Theorem 3.2. We explain Definition 5.1 in Subsection 5.1. For C ∞ flux, we prove that Definition 5.1 is equivalent to Definition 2.4 by Theorem 5.1. We compare our new definition with some other classical definitions in Subsection 5.2. We show that all definitions of nonlinear flux are equivalent for analytical flux. 5.1. Nonlinear smooth flux. We introduce a definition of nonlinear C ∞ flux related to critical geometric optics expansions. When the compatibility conditions (3.5) are satisfied in Theorem 3.1, high frequency waves are smooth solutions of the conservation law (2.1). Furthermore, these conditions are optimal thanks to Theorem 4.1. What is the highest frequency waves solutions of the conservation law (2.1) which can be propagated as in Theorem 3.1? That is to say, what is the critical geometric optics for the conservation law (2.1)? Of course, the answer depends on the flux. Indeed, near the constant state u we can propagate waves with frequency ε−m , m > 1, if the set {a0 (u), a00 (u), · · · , a(m−1) (u)}⊥ is not reduced to {0}. Thus the maximal m occurs when {0} = {a0 (u), a00 (u), · · · , a(m) (u)}⊥ and {0} = 6 {a0 (u), a00 (u), · · · , a(m−1) (u)}⊥ . We now can write the following definition. Definition 5.1. [Nonlinear smooth flux] Let the flux F belong to C ∞ (R, Rd ) and I = [−M, M ]. The flux is said to be nonlinear on I if, for all u ∈ I, there exists m ∈ N∗ such that (5.1) rank{a0 (u), · · · , a(m) (u)} = d. Furthermore, the flux is said to be genuine nonlinear if m = d is enough in (5.1) for all u ∈ I. In fact, the non-linearity is a matter of the second derivatives of F, a0 = F00. Notice that m ≥ d. We need at least d vectors in (5.1) to span the space Rd. Thus the genuine nonlinear case is the strongest nonlinear case. The genuine nonlinear case was first stated in [5] (condition (2.8) and Lemma 2.5 p. 447 therein). The genuine nonlinear condition in the d dimensional case (5.2) det(a0 (u), a00 (u), · · · , a(d) (u)) 6= 0, ∀u ∈ I, 14 STÉPHANE JUNCA was also in [8], see condition (16) p. 84 therein. The simplest example of genuine nonlinear flux F with the velocity a was given in [5, 8, 2]: 2 u ud+1 2 d a(u) = (u, u , · · · , u ) with F(u) = ,··· ,. 2 d+1 Definition 5.1 is a generalization of the genuine nonlinear condition (5.2). Definition 5.1 is more explicit with following integers with I = [−M, M ]. (5.3) dF [u] = inf{k ≥ 1, rank{F00 (u), · · · , F(k+1) (u)} = d} (5.4) dF = sup\|u\|≤M dF [u] ∈ {d, d + 1, · · · } ∪ {+∞}. ≥ d, Indeed, Definition 5.1 states that the flux is genuine nonlinear when dF reaches its minimal value, dF = d. Conversely, when the flux F is linear, a is a constant vector in Rd and dF reaches its maximal value, dF = +∞. Between dF = d and dF = +∞, there is a large variety of nonlinear flux. The following theorem gives the optimal parameter α (2.3) for smooth flux. Notice that this theorem is essentially a corollary of the uniform estimates given by Stein [30] for stationnary phase lemmas, see also [19]. Theorem 5.1. [Sharp measurement of the flux non-linearity ] Let F be a smooth flux, F ∈ C ∞ ([−M, M ], Rd ). Then, the measurement of the flux non-linearity αsup is given by αsup = 1 1. ≤ dF d Furthermore, when αsup > 0 there exists u ∈ [−M, M ] such that dF = dF [u]. A similar result for the genuine nonlinear case, dF = d, can be found in [2]. This theorem is a powerful tool to compute the parameter αsup , for instance: • F (u) = (cos(u), sin(u)) is genuine nonlinear flux , αsup = 1/2 since det(F 00 (u), F 000 (u)) = 1. • When F is polynomial with degree less or equal to the space dimension d, αsup = 0 and F does not satisfy Definition 5.1. • It is well known that the “Burgers multi-D” flux F (u) = (u2 , · · · , u2 ) is not nonlinear when d ≥ 2. Let us explain this fact by two arguments: the explicit computation of αsup and a sequence of high frequencies waves solutions of (2.1). – a00 (u) ≡ 0 so dF = +∞ and Theorem 5.1 yields αsup = 0. – The sequence of oscillations with large amplitude (uε )0<ε≤1 given s,1 2 blows up in any Wloc by uε (t, x) = uε0 (x) = sin x1 −x , s > 0: ε for all t, sup0<ε≤1 kuε (t, .)kW s,1 ([0,1]d ,R) = +∞. But the sequence of initial data is uniformly bounded in L∞ , kuε0 kL∞ = 1. Thus there is no improvement of the uniform initial Sobolev bounds. HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 15 • When F is polynomial such that deg(Fi ) = 1+i, F is genuine nonlinear: 1 αsup =. d Remark 5.1. For smooth Flux αsup is the inverse of an integer. Not all real value of αsup in [0, 1] are possible for F ∈ C ∞ . With less smooth flux, all other values of αsup are possible ([23, 13, 31, 19, 4, 3]). For sake of completeness, we give a proof of Theorem 5.1 related to stationnary phase lemmas ([30, 19, 2]). We mainly follow Stein [30]. Notice that, when Definition 2.4 is simplified by fixing τ = 0, a nice proof can be found in [19]. Our proof needs many lemmas. First we recall Lemma 1 p. 125 from [2] giving the optimal α for real functions. Lemma 5.1 ([2]). Let ϕ ∈ C ∞ ([−M, M ], R), mϕ [v] = inf{k ∈ N, ϕ(k) (v) 6= 0} mϕ = sup\|v\|≤M mϕ [v] ∈ N = N ∪ {+∞}, ∈ N, Z(ϕ, ε) = {v ∈ [−M, M ], \|ϕ(v)\| ≤ ε}. If 0 < mϕ < +∞ then there exists C > 1 dependent of the function φ such that, for all ε ∈]0, 1], 1 (5.5). C −1 εα ≤ meas(Z(ϕ, ε)) ≤ Cεα with α = mϕ To compute the measure of Z(ϕ, ε) with a different assumption, we adapt a proof of E. Stein about stationary phase method [30]. The main point in the following lemma is that the constant does not depend on the function φ. Indeed, the condition 1 ≤ \|φ(k) (v)\| is stronger than the condition mϕ = k. The following lemma is fundamental to prove Theorem 5.1. Lemma 5.2. [2] Let k ≥ 1, I an interval of R, φ ∈ C k (I, R). If then 1 ≤ \|φ(k) (v)\|, measure{v ∈ I, \|φ(v)\| ≤ ε} ≤ ck ε1/k , for all v ∈ I, where ck are constant independent of φ. Proof : Since the result is independent of the interval I and the constant sign of the derivative φ(k) on the interval, let us suppose that I = R and φ(k) (v) ≥ 1 for all v ∈ R. Thus we have for all v ≥ u : φ(k−1) (v) − φ(k−1) (u) ≥ v − u. This inequality shows that the function φ(k−1) admits an unique root. Assume φ(k−1) (0) = 0 without loss of generality. With these assumptions we prove the lemma when k = 1. Since \|φ(v)\| ≥ \|v\| for all v, we have Z(φ, I, ε) = {v ∈ I, \|φ(v)\| ≤ ε} ⊂ [−ε, ε]. So the lemma is proved for k = 1 with c1 = 2. We now prove the Lemma by induction on k > 1. We have for all v, \|φ(k−1) (v)\| ≥ \|v\|. Let η > 0 . Notice that meas(Z(φ, [−η, η], ε)) ≤ 2η. Let 16 STÉPHANE JUNCA ψ be the function φ/η. Notice that ψ (k−1) (v) ≥ 1 on ]η, +∞[. By our inductive hypothesis on ψ we have meas(Z(ψ, ]η, +∞[, ε) ≤ ck−1 (ε)1/(k−1) , so meas(Z(φ, ]η, +∞[, ε) ≤ ck−1 (ε/η)1/(k−1). A similar argument yields meas(Z(φ, ] − ∞, −η[, ε) ≤ ck−1 (ε/η)1/(k−1) . These previous three bounds give meas(Z(φ, R, ε)) ≤ g(η) = 2 η + ck−1 (ε/η)1/(k−1). This last inequality is valid for all η > 0. It suffices to minimize the function g on ]0, +∞[. A computation of the minimum yields meas(Z(φ, R, ε)) ≤ ck ε1/k , where ck = 4 (ck−1 /(k − 1))(k−1)/k which concludes the proof. The previous lemma is generalized to parameters in a compact set, see Lemma 4 p. 127 in [2]. Lemma 5.3 ([2]). Let P be a compact set of parameters, k a positive integer, A > 0, V = [−A, A], K = V × P , φ(v; p) ∈ C 0 (P, C k (V, R)), such that, for all (v, p) in the compact K, we have k X ∂j φ j=1 ∂v j (v; p) > 0. Let Z(φ(.; p), ε) = {v ∈ V, \|φ(v; p)\| ≤ ε}. Then, there exists a constant C such that supp∈P meas(Z(φ(.; p), ε)) ≤ Cε1/k. We now turn to the key integer dF. Lemma 5.4. If F is a nonlinear flux on I in the sense of Definition 5.1 then dF is finite and there exists u ∈ I such that dF = dF [u]. Proof. Let u be fixed in I. Then there exist, 1 ≤ j1 < j2 < · · · < jd = dF [u] such that rank{a(j1 ) (u), · · · , a(jd ) (u)} = d by the definition of dF [u]. So the continuous function g(v) = det(a(j1 ) (v), · · · , a(jd ) (v)) does not vanish at v = u. By continuity, this is still true on an open set J with u ∈ J. Since jd = dF [u], we have dF [v] ≤ dF [u] for all v ∈ J. Thus v 7→ dF [v] is upper semi-continuous and the result follows immediately on the compact set I. Now we are able to prove Theorem 5.1. Proof of Theorem 5.1. There are two steps. 1 step 1: αsup ≥. dF Set φ(v; τ, ξ) = τ + a(v) · ξ with τ 2 + \|ξ\|2 = 1. Since φ(.; τ, 0) = τ has no root, we can assume that ξ 6= 0Rd . For j ≥ 1 we have ∂vj φ(v; τ, ξ) = a(j) (v) · ξ. By definition of dF [v] there exists j ≤ dF [v] ≤ dF such that ∂vj φ(v; τ, ξ) 6= 0. Thus, we have when ξ 6= 0 (5.6) dF X j=1 \|∂vj φ(v; τ, ξ)\| > 0. HIGH FREQUENCIES AND REGULARITY FOR CONSERVATION LAWS 17 When ξ = 0, we have τ = ±1 since τ 2 + \|ξ\|2 = 1. The function φ(v; ±1, 0) = ±1 6= 0. By continuity of this function there exists an open neighborhood V of (1, 0Rd ) such the function does not vanish on V . Set P be the complementary set of V in the unit sphere of Rd+1 . P is compact and (5.6) is true on P . Now we can use Lemma 5.3 to conclude the first step. 1 step 2: αsup ≤. dF This inequality is already stated in [18, 19]. Let us give a proof to be selfcontained. u is defined in Lemma 5.4. Then, by the definition of u, there exists ξ ∗ 6= 0 such that ∂vj φ(u; τ, ξ ∗ ) 6= 0 for 1 ≤ j < dF and ∂vj φ(u; τ, ξ ∗ ) = 0 for j = dF . Notice that the previous derivatives are independent of τ . Let τ ∗ be −a(u) · ξ ∗ , so φ(u; τ ∗ , ξ ∗ ) = 0. To be on the unit circle we set τ = r τ ∗ and ξ = r ξ ∗ where r = ((τ ∗ )2 + \|ξ ∗ \|2 )−1/2 . ϕ(v) = φ(v; τ , ξ) vanishes at v = u and mϕ [u] = dF . Furthermore, since u is the point where the flux reaches its maximal degeneracy we also have mϕ = mϕ [u] = dF . Now, by Lemma 5.1, more precisely the first inequality of (5.5), and the definition of α (2.3), we have 1/mϕ ≥ α. Thus the second step is proved. Finally 1 1 ≤ αsup ≤ and the proof is complete with Lemma 5.4. dF dF 5.2. Comparisons with other nonlinear flux definitions. There are more general definitions of nonlinear flux [15, 23]. Definitions 5.2, 5.3 below are qualitative. We use quantitative definitions 2.4, 5.1. Indeed, the precise smoothing is related to Definition 2.4 or Definition 5.1 and the parameter αsup or equivalently dF . Let us compare theses definitions with Definition 5.1. It can be useful for other applications. Notice also that definitions depend on the flux regularity, but, for our purpose, it is not an important point. For C 1 flux, Lions, Perthame and Tadmor introduced Definition 2.4 and a more general definition of nonlinear flux, Definition 5.2. For C ∞ flux, we introduce Definition 5.1 related to stationary phase assumption on the flux. Indeed, by Theorem 5.1, Definition 5.1 is equivalent to Definition 2.4 for C ∞ flux. For C 2 flux, Definition 5.3 related to the second derivative of the flux is more general than Definition 5.1, even in the context of C ∞ flux. Finally, for analytic flux, all these definitions are equivalent. For less smooth flux we refer to the works of E. Yu. Panov ([25, 26]). Let us state Definitions 5.2 and 5.3 and prove our previous comparisons with Definition 5.1. Definition 5.2. [General Nonlinear Flux [23]] A flux F, differentiable on [−M, M ] is said to be nonlinear if the degeneracy set W (τ, ξ) = {\|v\| ≤ M, τ + F0 (v) · ξ = 0} has null Lebesgue measure for all (τ, ξ) on the sphere. 18 STÉPHANE JUNCA This definition is of a great importance since this condition implies the compactness of the semi-group St associated with the conservation law (2.1).	9,526
https://openalex.org/W4233729675	OpenAlex	Open Science	CC-By	2,020	Perception of Childbirth Experiences of Japanese Women in Bali, Indonesia: A Qualitative Study	Kazuko Tanaka	English	Spoken	10,617	16,906	Perception of childbirth experiences of Japane Indonesia: A qualitative study Kazuko Tanaka (  tkazuko@n.ypu.jp ) Yamaguchi Kenritsu Daigaku Kango Eiyo Gakubu https://orcid.org/0000-0002-8292-4930 Perception of childbirth experiences of Japanese women in Bali, Indonesia: A qualitative study Kazuko Tanaka (  tkazuko@n.ypu.jp ) Yamaguchi Kenritsu Daigaku Kango Eiyo Gakubu https://orcid.org/0000-0002-8292-4930 Ni Made Dian Kurniasari Universitas Udayana Kampus Surdiman Desak Nyoman Widyanthini Universitas Udayana Kampus Surdiman Ni Luh Putu Suariyani Universitas Udayana Kampus Surdiman Rina Listyowati Universitas Udayana Kampus Surdiman Akimi Urayama Yamaguchi Kenritsu Daigaku I Made Ady Wirawan Universitas Udayana Kampus Surdiman Koichi Yoshimura Yamaguchi Kenritsu Daigaku Research article Keywords: childbirth experiences, Japanese, pregnant women, Bali Indonesia, midwifery, qualitative study, woman-centred care Posted Date: December 8th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-30084/v3 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Perception of childbirth experiences of Japanese women in Bali, Indonesia: A qualitative study Kazuko Tanaka (  tkazuko@n.ypu.jp ) Yamaguchi Kenritsu Daigaku Kango Eiyo Gakubu https://orcid.org/0000-0002-8292-4930 Ni Made Dian Kurniasari Universitas Udayana Kampus Surdiman Desak Nyoman Widyanthini Universitas Udayana Kampus Surdiman Ni Luh Putu Suariyani Universitas Udayana Kampus Surdiman Rina Listyowati Universitas Udayana Kampus Surdiman Akimi Urayama Yamaguchi Kenritsu Daigaku I Made Ady Wirawan Universitas Udayana Kampus Surdiman Koichi Yoshimura Yamaguchi Kenritsu Daigaku Research article Keywords: childbirth experiences, Japanese, pregnant women, Bali Indonesia, midwifery, qualitative study, woman-centred care Posted Date: December 8th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-30084/v3 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Background Indonesia’s maternal mortality ratio was 446 per 100,000 live births in 1990 however, as a result of government efforts to improve maternal health services, by 2015 this ration had fallen to approximately 126 per 100,000 live births [1]. However, perinatal care including the maternal mortality ratio in Indonesia still has room for improvement. Previous Indonesian studies indicated that midwives did not have enough clinical competencies and skills particularly in anaemia management during pregnancy [2] and that many pregnant women were unsatisfied with antenatal services provided by midwives [3]. Patient satisfaction can be defined as patients’ judgements regarding the quality of care they have received [4]. Women assess the quality of perinatal care received based on their satisfaction with the services provided, thus influencing their utilization of the available health facilities [5]. Measuring women’s childbirth satisfaction is complex and multidimensional; however, determining women’s satisfaction provides essential and cost-effective feedback that contributes to improving institutional childbirth services [6]. In the current study, we focused on assessing womens’ satisfaction with quality of care given by midwives as described in the document “International Confederation of Midwives (ICM) Essential Competencies for Midwifery Practice” [7]. A midwife is described as a responsible and accountable professional who works in partnership with women and gives the necessary support, care and advice during pregnancy, labour and the postpartum period, to conduct births on the midwife’s own responsibility and to provide care for the newborn and the infant [8]. All women, including Japanese, generally request and expect support from midwives during pregnancy, birth, and the postpartum period [9-13]. Many foreigners live in Indonesia, and according to Japan’s Ministry of Foreign Affairs, the number of Japanese residents in Indonesia is increasing, particularly in Bali [14]. The Japanese community in Bali is demographically young and is experiencing a baby boom [15]; these residents are predominantly women in their 30s and 40s living there with their husbands and children. Migrants around the world generally tend to have negative experiences with maternal care in their new countries. Previous studies have reported that migrant women in developing countries usually do not receive timely, appropriate or high-quality maternal health care [16-18]. A systematic and comparative review of immigrant and non-immigrant women’s experiences of maternity care, revealed that immigrant women were less positive about the care they received than non-immigrant women because of reasons such as communication problems and lack of familiarity with care systems [19]. Research article License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License k is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published on December 7th, 2020. See the published version at https://doi.org/10.1186/s12884-020-03466-x. Page 1/17 Page 1/17 Abstract Background: Maternal healthcare services in Indonesia have seen dramatic improvements over the past 25 years and yet there is still room for improvement. The perception, by the women, of the perinatal care provided, is a vital input to further improving these services. This study examines how the perinatal care provided is experienced by Japanese women in Bali, using an interview survey. Methods: We conducted semi-structured interviews, from August to October 2017, with 14 Japanese women living in Badung Regency and Denpasar City in Bali Province, Indonesia to report their perception of the perinatal care they experienced during their pregnancies. The interview guide included among others, the reasons for choosing specific (perinatal care) health facilities and their satisfaction with their experience of using the antenatal, delivery, and postnatal care services. The data were analysed using the qualitative content analysis method. Results: From the interview data, 12 categories across five themes were extracted. Participants reported experiencing various concerns during their pregnancies such as difficulty in obtaining perinatal care related information. From the beginning of their pregnancies, participants gradually established trusting relationships with midwives, but in many situations, they were disappointed with their childbirth experiences, as they felt that the care provided was not woman-centred. Through their own efforts and with the support of family members and other Japanese residents, many women were able to eventually regard their childbirth experiences as positive. Nevertheless, some women could not overcome their negative impressions even years after childbirth. Conclusions: Participants desired close attention and encouragement from nurses and midwives. Our results suggest that Japanese women in Bali expected a woman-centred perinatal care and active support from nursing/midwifery staff during their pregnancies and postnatal care. Design Our study employed a qualitative descriptive design using in-depth semi-structured interviews. Qualitative description represents the methodological category with the least level of inference among the qualitative methods [20], and in-depth semi-structured interview data constitutes the empirical backbone of qualitative research in the social sciences [21]. The theoretical framework that guided this study was a derived Donabedian's model [22]. According to this model, quality of care is drawn from three categories: structure, process, and outcome. We used this model to understand childbirth experience and evaluate midwives’ quality of care, which allowed insight into mothers’ satisfaction during the perinatal period. To assess the quality of midwifery care, we also referred to Halldorsdottir and Karlsdottir’s theory [23] and the concept of woman-centred care [24]. Halldorsdottir and Karlsdottir highlight five principal factors in a midwife’s profession. These five principal factors are: professional caring, competence and wisdom, empowering interaction and partnership together with the midwife’s personal and professional development holistically combined [23]. In order to focus our objective, we asked participants about their childbirth experiences, their satisfaction or dissatisfaction with the attitudes of, and services provided by midwives during the perinatal period. The consolidated criteria for reporting qualitative research (COREQ) checklist was used to guide reporting of this study. The COREQ checklist contains 32 items [25]. empowering interaction and partnership together with the midwife’s personal and professional development holistically combined [23]. In order to focus our objective, we asked participants about their childbirth experiences, their satisfaction or dissatisfaction with the attitudes of, and services provided by midwives during the perinatal period. The consolidated criteria for reporting qualitative research (COREQ) checklist was used to guide reporting of this study. The COREQ checklist contains 32 items [25]. Qualitative research is based on interpretation, which requires input from researchers. Although interpretation is subjective, the authors of this study are well qualified to understand the nuances of the topic. The first author (KT) is a female Japanese midwifery lecturer in a university who has had midwifery experience in Japan and Southeast Asia. From her personal experience, the first author felt there was room for improvement in perinatal care in Southeast Asia, especially in midwifery care during birth. All Indonesian co- authors are public health and maternal and child health research experts (NMDK, DNW, NLPS, RL, IMAW) and university lecturers. Setting Bali is renowned worldwide as a popular tourist destination and has many foreign residents and immigrants from other islands in Indonesia. Bali is one of the most developed islands in Indonesia in terms of economic activity, infrastructure, and population [26]. The number of Japanese residents in Bali is nearly 40 times greater than it was 30 years ago. At the time of this study, a Japanese Foreign Ministry survey showed that there were approximately 3,000 Japanese nationals residing in Bali [14]. The data for this research were collected from the Badung Regency and Denpasar City (the province’s capital) in Bali Province, where three quarters of these Japanese residents in Bali were located [14]. Design As for the Japanese co-authors, AU is a female professor in the division of midwifery and KY is a male medical doctor and professor in the Graduate School Background Communication problems and lack of familiarity with health care systems had a negative impact on immigrant women’s experiences, as did perceptions of lack of respect, kindness and discrimination in the care their received, [19]. Non-native and immigrant women, including the Japanese living in Indonesia may have Page 2/17 Page 2/17 differing expectations and perceptions of the maternal care during their pregnancy and, as a result, they may be raising new issues and concerns that could be considered for further improving maternal and nursing care in Bali, Indonesia. differing expectations and perceptions of the maternal care during their pregnancy and, as a result, they may be raising new issues and concerns that could be considered for further improving maternal and nursing care in Bali, Indonesia. There is little research in Indonesia that has focused on maternal satisfaction, expectations or experiences of native and migrant women during the perinatal period. In our study we intended to explore, through respondent interviews, the perinatal experiences of the migrant Japanese women residing in Bali. Because many of these women could compare perinatal care in Indonesia with what is provided in Japan, we expected that they would identify more satisfaction and experience related issues than the local Balinese women. Data collection Their answers, we concluded, could provide insights that would assist the local and Japanese interviewers to look at any similarities and differences in their responses and types of issues raised. Semi-structured, face-to-face interviews were conducted with the 14 participants to explore their explanations, perceptions and experiences with the perinatal midwifery care they received during their childbirth experiences. Before starting the interviews, consent and background information was obtained, including age, religion and duration of stay in Bali, length of the marriage, reproductive history, and Indonesian-language proficiency (Additional file 2). In our interview guide we included questions on reasons for choosing a particular childbirth facility and their explanations, perceptions and experiences (both positive and negative) about their midwives’ attitudes and services they received during the perinatal period, including their knowledge and skills, interpersonal communication abilities, partnering with women and their families, and so on (Additional file 1). All of the participating women consented to being interviewed which were conducted in the participants’ homes or at other mutually agreed places where privacy could be guaranteed. The interviews were audio recorded and the participants’ consent and contemporaneous notes were also taken. The first author (KT) conducted the semi-structured face-to-face interviews in Japanese. The Indonesian researchers also participated in the interviews taking field notes with the support of a professional female Japanese interpreter who was familiar with local medical terminologies and conditions. The interpreter received advance orientation about the aim of the study and its themes. Each interview was jointly conducted in Japanese by the first author and an Indonesian researcher supported by the interpreter to enable multiple observations and conclusions. Each interview lasted approximately one hour. At the end of the interview, the interviewer (KT) verbally summarized the key points and asked the participant if the summary was accurate. Data collection was characterised by openness to new ideas among the interviewers and reinforced by the follow-up probing questions during these interviews. Theoretical saturation was achieved after 14 interviews as no more new information emerged. Sample recruitment For the purpose of this study, Japanese women, aged 26-42 years, who had given birth in Bali, were recruited. Every woman who consented to participate in the study had to meet the criteria of having their youngest child to be under five. All of these women had knowledge of and many had experienced maternity care services in Japan. Only 3 women from these recruited women had never given birth before. Nevertheless, based on their knowledge, the authors concluded that they could still compare their perinatal experiences in Indonesia with those in Japan. The first participant was introduced to the researchers by a Japanese permanent resident working in Bali. Through the snowball sampling method, a total of only 14 participants were recruited. The researchers collected data between August and October 2017. Page 3/17 Page 3/17 Page 3/17 Data collection The quality of midwifery services plays a decisive role in a woman’s experience of childbirth [23], and because recent work in policy and service provision surrounding maternity care recommends a woman-centred approach [27,28], we developed an interview guide for this study based on the notion of a midwife's professionalism [23] linked to the concept of woman-centred care [24]. The Japanese researchers (KT, AU, KY) designed the first draft of the interview guide and then consulted about it with Indonesian researchers (NMDK, DNW, NLPS, IMAW) (Additional file 1). The interview guide was tested on the local Balinese women to gain insight into what sort of satisfaction and services related issues that may be raised by these women using these interview questions. Coincidentally, there was similar research being conducted at the same time on Balinese women by our research group and so we decided to use the same interview guide for interviewing the Japanese in light of the limited time we had for the study. Their answers, we concluded, could provide insights that would assist the local and Japanese interviewers to look at any similarities and differences in their responses and types of issues raised. The quality of midwifery services plays a decisive role in a woman’s experience of childbirth [23], and because recent work in policy and service provision surrounding maternity care recommends a woman-centred approach [27,28], we developed an interview guide for this study based on the notion of a midwife's professionalism [23] linked to the concept of woman-centred care [24]. The Japanese researchers (KT, AU, KY) designed the first draft of the interview guide and then consulted about it with Indonesian researchers (NMDK, DNW, NLPS, IMAW) (Additional file 1). The interview guide was tested on the local Balinese women to gain insight into what sort of satisfaction and services related issues that may be raised by these women using these interview questions. Coincidentally there was similar research being conducted at the same time on Balinese women by our research group and so we Coincidentally, there was similar research being conducted at the same time on Balinese women by our research group and so we decided to use the same interview guide for interviewing the Japanese in light of the limited time we had for the study. Participant Characteristics The participants’ average age was 37.6 (± 4.7) years. Of the 14 participants, three had a high-school education and 11 had tertiary education. The average length of stay in Bali was 8.8 (± 5.1) years; the majority 10(71%) had been in Bali for over five years. Most of the women’s husbands were Indonesian. {10(71%) were Indonesian, 4(29%) were Japanese}. Six participants were Muslims, three were Hindu, (they were required to convert when they got married with Indonesian men) and five were adherents of other religions. Over half of the participants could read and write in Indonesian. Of the 14 participants, 11 had given birth two or more times, and six of the 14 women had also experienced childbirth in Japan. In terms of the choice of birthing facilities, 11 participants had chosen private hospitals and three had chosen a midwives’ clinic (bidan in Indonesian). Six women had given birth vaginally, two had been induced, five had had caesarean sections, and one had experienced a waterbirth. All babies were full-term. The average age of their youngest child was 21 (± 15) months. Only three participants possessed the Indonesian government’s Mother and Child Health (MCH) Handbook, while over half of the women retained the Japanese government’s MCH Handbook (Table 1). Ethical considerations Researchers adhered to the Declaration of Helsinki recommendations in conducting this research. Ethical approval was obtained from the Faculty of Medicine, Udayana University/Sanglah Hospital (493/UN.14.2/KEP/2017), and Yamaguchi Prefectural University (No.27-45). Before data collection, the purpose and aim of the research were explained to the participants, and they were informed about their rights. Written and verbal information was given to participants, and written consent was obtained before the interview. Participants were also assured that all their responses would be kept confidential; all collected information including audio recordings and transcripts were securely stored and accessible only to the research team. This study is a part of an ongoing project (2016–2021), which was implemented in cooperation with Udayana University and Yamaguchi Prefectural University. Data analysis To ensure reliability, field notes and individual interview data were compared to intensively examine the consistency of data [36, 37]. The ATLAS.ti 8 software package was used to support analysis of the transcripts. compared the categories and themes generated from the Japanese and English data sets [34]. Categories and themes were refined, reviewed and reconciled going back to the data repeatedly to ensure consistency. Triangulation was used to reduce the effect of research bias to establish confirmability, including reflexive field notes. Interview data, data analysis products, and data reconstruction products were provided to the research team for verification. To ensure reliability, field notes and individual interview data were compared to intensively examine the consistency of data [36, 37]. The ATLAS.ti 8 software package was used to support analysis of the transcripts. Data analysis The data were independently analysed using the qualitative content analysis in accordance with Graneheim and Lundman’s methods [29]. Content analysis is defined as a research technique for identifying replicable and valid inferences from data in context [30]; it is a flexible method for analysing textual data [31]. This method can be useful as one stage of data analysis because it allows the relevance of a pre-existing theory to be tested, and it can be used as a way of assessing the applicability of theory [32]. The process includes open coding, category creation, and abstraction [33]. After completing the interviews, the Indonesian and Japanese researchers (KT and NMDK, DNW, or NLPS) involved in conducting the interviews reviewed and discussed the childbirth experiences that had been described by the participants. Recordings were transcribed verbatim into Japanese by the first author and later translated into English by one professional translator. We used only one translator to ensure the reliability of the data [34]. English transcripts were given to the Indonesian researchers (NMDK, DNW, NLPS) who checked it and compared it with their field notes. The data analysis process was led by the Japanese researchers (KT, AU, KY). All Japanese researchers and Indonesian researchers (NMDK, DNW, NLPS) repeatedly read the verbatim records in order to familiarise themselves with the participants’ expressions, and then extracted and summarised related text in order to avoid changing the individual participants’ overall meanings. Researchers performed the analysis separately and then discussed the results as a team and arrived at a consensus [30, 35]. The five phases of analysis [33] began with the researchers familiarising themselves with the data and identifying the preliminary codes. The codes were integrated into subcategories until the main categories emerged. This process allowed us to identify key comprehensive themes that described the investigated phenomena, as well as the key elements of interest. We developed categories and themes, then Page 4/17 Page 4/17 compared the categories and themes generated from the Japanese and English data sets [34]. Categories and themes were refined, reviewed and reconciled going back to the data repeatedly to ensure consistency. Triangulation was used to reduce the effect of research bias to establish confirmability, including reflexive field notes. Interview data, data analysis products, and data reconstruction products were provided to the research team for verification. Themes and Categories When the participants were preparing to give birth in Bali, they desired a safe pregnancy and childbirth experience. They had a variety of concerns and worries regarding giving birth at an older age or fears of miscarriage and unpredictability in the onset of labour, and they wanted a high level of maternity care. Consequently, they placed high importance on finding specialist obstetricians and well- equipped health facilities where they could undergo maternity related ultrasound examinations. These women also preferred natural childbirth, to give birth with their husbands in attendance, doula support, kangaroo mother care, and/or exclusive breastfeeding. As a result, they selected childbirth facilities that could provide these services. In effect, these women were expressing the sort of childbirth plans they preferred. A Japanese friend who had been a midwifery professional said that she would be present for the birth, even if it was in the middle of the night. Also, I kept hearing that hospitals here perform a lot of caesarean sections, so I asked them to perform a caesarean section only if the situation became very bad and with my consent, but, if there were no problems, I said that I would prefer to have natural childbirth. The doctor said that it was his policy was to proceed with a natural childbirth whenever possible (JPN5). After becoming pregnant, the Japanese women searched for an obstetrician they could trust, underwent regular health check-ups, and then gave birth at the hospital or clinic where their attending obstetrician worked. The women who chose to give birth in midwives’ clinics reported developing trust in their midwives because of the midwives’ attitudes, good listening skills, and their efforts to help the birth occur in a home-like atmosphere, and also because they provided advance explanations of the procedures that were to be undertaken. The (bidan) staff were wonderful people. They always smiled when they spoke, and if I had diarrhoea when I visited, they would give me lots of advice […], so I kind of depended on them (JPN6). Almost three-quarters (71%) of the Japanese women’s spouses were Indonesian, and they were based in Bali; consequently, they chose to give birth in Bali rather than travel back to Japan despite their anxiety. Themes and Categories Some women sought a feeling of religious solidarity and chose care providers of the same religious faith, while others sought a sense of security and peace of mind at accessible midwives’ clinics (bidan in Indonesian). Women with no experience of childbirth found greater peace of mind in giving birth in familiar surroundings. Moreover, they were grateful that they could, without feeling guilt or embarrassment, ask staff to look after their infants after birth, a common practice in Bali, unlike in Japan. I heard in Japan that if you asked hospital staff to look after the baby at night, they would make a stern face, as if to say: ‘No, you’re the mother, you look after it’. Friends told me that staff would refuse to look after the baby, saying things like, ‘It’s born now; that is the mother’s job’, but in Indonesia, this isn’t the case. I’ve heard that in Japan, the nurses make you feel really guilty (JPN5). Themes and Categories We identified 12 categories across five themes, which are described below (Figure 1). During their perinatal period, the participants strongly emphasised safe and satisfying childbirth experiences (Figure 1: I-1). However, they had various worries and concerns regarding issues such as the difficulty of obtaining information pertaining to pregnancy and childbirth (Figure 1: II-1). For this reason, support from family members and Japanese friends who had previously worked as midwives and nurses in Japan, was essential from the beginning of their pregnancy (Figure 1: II-3). Although the participants did not have high expectations of the midwives caring for them (Figure 1: II-2), they did seek comfort and peace of mind (Figure 1: I-3). From the beginning of their pregnancies, they gradually built relationships of trust with the midwives or other caregivers (Figure 1: I-2), but many were disappointed with their childbirth experiences, as they felt that the care provided during labour was not centred on the woman giving birth (Figure 1: III-1). However, through both their own efforts (Figure 1: IV-1) and with the support of family members and the Japanese friends as mentioned earlier (Figure 1: II-3), many women were able to reflect on their childbirth experience positively (Figure 1: IV-2). However, some women could not achieve this, even years after they had given birth (Figure 1: III-2). Additionally, empowerment of mothers during pregnancy, childbirth, and the postpartum confinement was regarded as important (Figure 1: V-1), and participants expressed their hope that perinatal care in Bali, Indonesia would further improve in the future (Figure 1: V-2). Desire for a satisfying childbirth experience Page 5/17 Page 5/17 When the participants were preparing to give birth in Bali, they desired a safe pregnancy and childbirth experience. They had a variety of concerns and worries regarding giving birth at an older age or fears of miscarriage and unpredictability in the onset of labour, and they wanted a high level of maternity care. Consequently, they placed high importance on finding specialist obstetricians and well- equipped health facilities where they could undergo maternity related ultrasound examinations. These women also preferred natural childbirth, to give birth with their husbands in attendance, doula support, kangaroo mother care, and/or exclusive breastfeeding. As a result, they selected childbirth facilities that could provide these services. In effect, these women were expressing the sort of childbirth plans they preferred. Embarrassment and disappointment During pregnancy check-ups in hospital by the obstetricians, the Japanese women had virtually no interaction with midwives. Having the belief that health guidance comes from the midwives, based on their knowledge of Japan, this lack of health guidance made them feel anxious. During childbirth, doctors directly assisted with the labour; so the women did not remember any nurses or midwives being present in the birthing room. Comparing this with the situation in Japan, where midwives help during labour, it was understandable the women sensed the extensive authority of the physicians and felt that midwives had an unclear role. Because of this comparison, they were especially discomfited by light level of midwifery and nursing care during labour and birth. When the women entered the hospital with labour pains or after their membrane ruptured midwives checked on them in accordance with their local standards and, since they did not receive clear advice regarding alleviating childbirth pain or actions they should take during labour when compared to their knowledge and experience in Japan, participants felt abandoned and also experienced distress because they perceived that they did not receive the expected support during labour. Many participants were especially anxious about breastfeeding, unsure about how to treat the umbilical cord when bathing the baby, and they expressed concern about limited guidance provided. Even when the women had the support of family members or friends, they still expected additional health based guidance from professionals. Despite planning for a vaginal birth, women did not feel that they were given enough support from the midwives regarding education on pain management during birth, guidance on breastfeeding, bathing the baby, and so on. Women who had previously experienced childbirth in Japan perceived significant differences in maternity care provided in Bali when compared to Japan that became a source of concern for them. The most memorable episode was that when I was giving birth in Japan to my first child, a midwife supported me during labour to avoid push with each contraction, but I didn’t have such specific support here (in Bali) (JPN1). If the staff had knowledge, I wish they had shared it with me. Concerns about local maternity services Consequently, even when the level of care they received differed from what they hoped for, they convinced themselves that it could not be helped. Having no expectations for maternity care is the best way not to get hurt, and my attitude is the same toward some of the other local people. After living here for seven years, you get good at it. But don’t get me wrong, Indonesian people are good people; I love them (JPN3). Many of the participants had networks of Japanese people they could consult. They were aware of the importance of support from their family and Japanese friends who had been midwives and who also worked in Indonesia. When they, the respondents gave birth, their mothers arrived from Japan to assist them, and Japanese friends who had been midwives living in Indonesia provided support until the postpartum confinement period ended. For women with Indonesian husbands, there was, on one hand, the added hardship of receiving a large number of relatives who visited after the birth but, on the other hand, family members also provided essential assistance, such as support for breastfeeding and various other aspects of child-raising. Ms. A. (Japanese friend who had been a midwife) gave me detailed explanations, and I was able to consult Ms. A. about even trivial matters. She was that kind of person. Accordingly, I was not anxious in the least, and it was helpful that she stayed with me during the birth as well (JPN2). Concerns about local maternity services Even participants who had no difficulty engaging in everyday conversations in Indonesian had some trouble communicating with midwives, as a result of unfamiliarity with specialist terminology, and many participants reported receiving support from Japanese friends who had also been midwives and other similar health-care professionals who worked in Indonesia. The women experienced difficulty obtaining information related to local maternal and child health services and facilities because of language and technical barriers and were worried that, as foreigners, they could be at a disadvantage. They feared that they would undergo an unnecessary caesarean section without their consent. Three women were given the Indonesian government’s MCH handbook. Indonesia’s maternal care is unlike Japan’s [2,38], where fourteen maternity check-ups are provided without charge. As this was not the case in Bali, participants felt they had less support provided to them. Consequently, the local maternity care system added to the participants’ unease regarding giving birth and other pregnancy-related difficulties. The women also felt anxious about the fewer number of examinations conducted during pregnancy in Bali when compared to Japan, and they had doubts regarding diagnoses and prescriptions. Additional differences in the maternity services between Bali and Japan also caused great concern, such as the short duration of postpartum hospital stays, generally one night and two days [39, 40]; when compared with the postpartum hospitalisation period in Japan of 5–7 days [41, 42]. They have many caesareans here. I would hate to have a caesarean. I wanted, if possible, to give birth naturally. Here, there is a tendency to quickly resort to surgery. All of the staff at my company gave birth via caesarean section (JPN3). Page 6/17 Page 6/17 I would like them to carry out a few more tests such as urine tests and blood pressure tests before determining that a C section was really necessary for the baby’s and the mother’s sake, (JPN9). I would like them to carry out a few more tests such as urine tests and blood pressure tests before determining that a C section was really necessary for the baby’s and the mother’s sake, (JPN9). They had adopted an attitude of generally not having high expectations during their social inter- personal encounters and this also extended to health staff, meaning they had fewer expectations from midwives from the beginning. Embarrassment and disappointment Rather than leaving the breastfeeding process after birth to the mother, I would have liked it if they had provided a little more guidance.…I thought that they probably don’t provide guidance for how to bathe the baby and every little thing because there are family members to do that (JPN9). Those experiences left women dissatisfied with their experiences during and after birth, and hoped they could have better experiences in the future. Women who were unable to bear the labour pains and underwent caesarean sections felt regret that they could have perhaps borne the pain if they had received help and encouragement. Page 7/17 Page 7/17 I was in the throes of birth, I couldn’t cope with labour pain, I asked help from the midwife, but she just said ‘Not yet’. I felt increasingly sad and neglected because no one did anything for me during labour. Finally, I said, ‘Please perform caesarean’ (JPN8). I was in the throes of birth, I couldn’t cope with labour pain, I asked help from the midwife, but she just said ‘Not yet’. I felt increasingly sad and neglected because no one did anything for me during labour. Finally, I said, ‘Please perform caesarean’ (JPN8). Future hope At Japanese childbirth facilities, healthcare guidance is provided through mothering classes and individual instruction from midwives, but only a few of the participants received similar guidance in Bali. Many of the women studied these things on their own, using the internet and other resources. During pregnancy check-ups, they felt that information and explanations were not actively being provided to expectant and nursing mothers; for instance, regarding examination results, they were simply told if there were any problems but they were not given additional detailed information to reassure the mother. The women also reported that information was not offered on topics such as weight gain and foods to be avoided during pregnancy. The participants knew that excessive weight gain during pregnancy can increase the risk of gestational diabetes, macrosomia, and/or hypertensive disorders of pregnancy, and they felt that they needed to take the initiative to obtain relevant information. This was probably due to the fact that the advice given to the local Balinese pregnant may not be appropriate for the Japanese participants. The participants also thought that the care and limited information provided to the local expectant mothers by their doctors who were in charge of their maternity care as problematic when compared to their knowledge and experiences in Japan. This highlighted the need for childbirth preparation and education for foreign expectant women that can help broadens their options. My hands and feet were really swollen and puffy; my blood pressure was elevated. I was worried, so I did a search on the Internet, and I found quite a number of diseases/conditions I would probably have been diagnosed with in Japan, such as hypertensive disorders of pregnancy. That was my situation, but here I didn’t receive any particular explanation or advice (JPN5). In contrast to Japan, Indonesia has a high birth rate. Some participants thought that their obstetricians and gynaecologists would have a great deal of experience, and that there were also veteran midwives who had established their own practices. The participants wanted to receive care from experienced professionals. Some participants who experienced discomfort during pregnancy found relief after receiving treatment at midwives’ clinics, while other participants were pleased to receive breast massages after birth, which improved milk production. When I went to my husband’s parents’ after childbirth, I got breast massage from an elderly traditional midwife, it helped me with exclusive breast-feeding. Women’s potential strength After giving birth, many of the participants did not receive clear guidance about breastfeeding or caring for the infant and consequently taught themselves instead. This is the way it is culturally done in Bali because of the available family support structures. While they were in hospital, they watched internet videos and referred to Japanese leaflets and childcare books since many of these were in Japanese and therefore understandable. Furthermore, those who had experienced childbirth in Japan drew on their previous and existing knowledge of labour and child-rearing to use in their situation. They didn’t teach me anything about how to care for the baby, even how to change a diaper. […] I didn’t really expect the hospital staff to teach me these things, and so I read various (Japanese) books on child-raising. (JPN7). Even though some of the women were dissatisfied with various aspects of the perinatal care they received, they were relieved that their babies had been born healthy. They did not regret their decision to give birth in Bali and were able to regard their pregnancy and labour experiences as positive. I had really strong feelings about ‘What if it had been a natural birth’? but all of a sudden they decided I needed a caesarean and I just said something like, ‘OK, as long as the baby is born healthy’. Ultimately, I felt that it was more important that the baby was born healthy than whether I wanted to do this or that. And so, in the end, I was just happy that the baby was born healthy, and that we were able to leave the hospital and that the baby grew healthily (JPN11). Need for responsive and competent staff in the maternity care facilities In 2012, the rate of caesarean sections in Indonesia was 12% [45] but, according to earlier research, there was a disparity regarding this between maternity facilities in urban and rural areas, with rates being higher in more urbanised areas such as Java and Bali [26]. Caesarean section rates are continuing to rise, particularly in high- and middle-income countries [46, 47]. When medically justified, a timely caesarean section can effectively reduce maternal morbidity and mortality; however, there is no evidence of the benefits of caesarean birth for women or infants who do not require the procedure [48]. According to new research by Indonesian researchers, the rate of caesarean sections is 23.0% in the urban areas of Indonesia [49]. All participants were aware of the high rate of caesarean sections in Bali; therefore, this was a major factor increasing their anxiety (as they preferred to give birth naturally). Even though the foetus and mother were not in danger, some participants requested caesarean sections during the first stage of labour because they were unable to bear the pain of childbirth. These women strongly believed that they would have been able to withstand the pain with active support, encouragement and explanations about their diagnoses from their midwives and other health staff. Women who received continuous support during labour were more likely to give birth ‘spontaneously’; that is, vaginally without the use of ventouse, forceps, or a caesarean section. In addition, such women were also less likely to use pain medications and to feel more satisfied and have shorter labour [50]. Moreover, in Bali, with the support of their mothers and other family members, the women did their best to overcome their unsatisfactory experiences during childbirth. Mental health issues can easily arise during the early postpartum period [42,51,52]; although the majority of women coped well. Nevertheless, for some, childbirth was a stressful experience; this is of concern given that in previous work it has been reported that childbirth-related stress increases the potential for developing postpartum posttraumatic stress disorder symptoms [53]. Literature reveals that the prevalence of postnatal depression is highest among migrant women [54]. Brief midwife-led counselling interventions for women who report distressing birth experiences have been found to be effective for reducing symptoms of trauma, depression, stress, and coping with feelings of self-blame [55]. Future hope There are many midwives’ clinics here, they shared with us useful pieces of maternal health trivia, and they were friendly (JPN10). It may be that in Indonesia having children is almost too routine; pregnancy and childbirth are not treated as big and special events. But I think the process could be made a little more enjoyable. As a mother, there are only a limited number of times you can Page 8/17 Page 8/17 experience childbirth in your lifetime, aren’t there? That’s why I think there could be more opportunities for enjoying the childbirth process (JPN7). experience childbirth in your lifetime, aren’t there? That’s why I think there could be more opportunities for enjoying the childbirth process (JPN7). I think the care provided for childbirth is OK as far as locals are concerned, but I think that health-care professionals at health facilities where foreigners are treated need to provide more information and support about postpartum care and caring for pregnant women. (JPN13) Need for information Private hospitals are the most popular maternity facilities in Bali and Indonesia (used by 48% of women; [26]), and the substantial majority of our participants gave birth in private hospitals in Bali. When choosing a facility for prenatal check-ups and childbirth, the participants consulted their networks of Japanese people as well as local information. For foreign mothers generally, maternal and child health information in their native language is extremely limited and among our participants, few possessed the Indonesian version of the MCH Handbook which was adapted from Japan’s maternal and child health system [43]. The Handbook facilitates the continuum of care throughout pregnancy, birth, postpartum, and the child’s infancy, and thus has the potential to be an effective tool for improving health awareness and client-provider communication. It includes maternal and child health tools that record the maternity related information as well as child development milestones from 0 to 5 years [44]. However, in Bali, Indonesia, there were some private health facilities where no MCH Handbook was available. These were found mostly at government-owned public health centres. Thus improving the availability of the local MCH Handbook in all maternity health care facilities available to all mothers would strengthen the maternity care services. Conclusions This study shows that the perception of the childbirth process as experienced by Japanese women in Bali, Indonesia during the intrapartum and puerperium periods was generally not woman-centred. The selected Japanese women who were interviewed sought, but, in most cases, did not receive, active support and encouragement from midwives who they interacted with during their childbirth experiences. Our results highlight the need for providing a more woman-centred care approach that includes the empowerment of women generally and, Japanese women especially, during the perinatal/maternity period. In general terms, maternal health care in Indonesia has significantly and consistently improved over the past 25 years and further research from women’s perspectives, especially from foreign women, provide critical insights from the lessons learned, that could contribute to improving the quality of perinatal/maternal care in Bali in particular and, in Indonesia generally consistent with the evolving international standards, and yet responsive to local socio-cultural needs. Women were seeking clear and detailed explanations and an accommodating attitude by midwives during perinatal/maternity period. Taking action to improve maternal care services by training and providing specific practice guidelines, adequate interpersonal communication skills and providing sufficient and effective skill development training and health education materials to be used for counselling is required. To develop appropriate information education and communication materials which are easy-to-understand even for foreigners could positively improve the health education provided by midwives. In addition, to contribute to improving maternal health services, it is necessary for medical professionals such as midwives to form partnerships with women, respect them, and cooperate with them. We conclude that midwives need to respect women's choices so that women themselves can be enabled to face their pregnancy and childbirth constructively and safely and with reassurance, and build a system to help these women to take the initiative in childbirth. Woman-centred care After childbirth, all new mothers tend review their childbirth experiences and events, reflecting on how they differed from what they had expected, and assimilate this experience in line with their expectations [56]. Recently, in Japan, there has been an increase in the Page 9/17 Page 9/17 number of maternity facilities where women who had just given birth along with midwives, can review their labour experience during the early postpartum period [57, 58]. Some of our study participants were disillusioned with their childbirth experiences and continued to recall this disillusionment during their interviews. For such women to overcome their negative childbirth experiences, reviewing their childbirth experiences with a midwife during the early postpartum period would have been an effective measure to help with coping better. number of maternity facilities where women who had just given birth along with midwives, can review their labour experience during the early postpartum period [57, 58]. Some of our study participants were disillusioned with their childbirth experiences and continued to recall this disillusionment during their interviews. For such women to overcome their negative childbirth experiences, reviewing their childbirth experiences with a midwife during the early postpartum period would have been an effective measure to help with coping better. Over the past 20 years, there have been various studies mostly in UK and Japan on women-centred care during pregnancy and childbirth [24, 27, 28, 38, 59-62]. The qualitative and quantitative results of a previous survey of expectant and nursing mothers done in the UK indicated that there is a need for woman-centred care through which midwives form partnerships with expectant and nursing mothers, and help expectant and nursing mothers make information-based choices [63,64]. Many of our participants were unable to form such partnerships with their local midwives during the perinatal period, consequently were unable to achieve the desired childbirth experience. Our results suggest that empowering women through the provision of information and attentive care throughout the perinatal period would be a valuable improvement. To enable women to approach pregnancy and childbirth proactively, and to have safe and satisfying childbirth experiences, it is essential that they receive support from their midwives that could contribute to further improvements in perinatal care in Bali, Indonesia. Limitations In this study, only Japanese women living in Indonesia were selected as study participants because they could compare their experience of perinatal care provided both in their local area Bali and in Japan. This comparison could help reveal relevant issues relating to perinatal care by comparing maternity care services in Bali, Indonesia and Japan. Thus, women’s perceptions about their experiences are solely described from the viewpoint of Japanese women which limits the value of the comparison between their perception of the care in Bali Indonesia and their recollections of their care in Japan. In addition, our study participants all used private facilities for their maternity care during their perinatal period. In Indonesia, giving birth in private facilities has increased to 48.1%, but many other women still give birth in public health facilities and at home [26]. Authors' contributions KT: Conceptualization, Methodology, Formal analysis, Data Curation, Investigation, Resources, Writing - Original Draft, Funding acquisition NMDK: Methodology, Formal analysis, Investigation, Data Curation, Writing - Review & Editing DNW: Methodology, Formal analysis, Investigation, Writing - Review & Editing NLPS: Methodology, Formal analysis, Investigation, Writing - Review & Editing RL: Validation, Writing - Review & Editing AU: Methodology, Formal analysis IMAW: Conceptualization, Methodology, Supervision KY: Conceptualization, Methodology, Formal analysis, Writing - Review & Editing, Supervision All authors have read and approved the manuscript. Ethics approval and consent to participate Ethics approval and consent to participate Ethical approval was obtained from the Faculty of Medicine, Udayana University/Sanglah Hospital (493/UN.14.2/KEP/2017), and Yamaguchi Prefectural University (No.27-45). Written and verbal information was given to participants, and written consent was collected before the interview. Abbreviations ICM: International Confederation of Midwives; COREQ: Consolidated criteria for reporting qualitative research; MCH: Maternal and Child Health; UK: United Kingdom Page 10/17 Page 10/17 Page 10/17 Acknowledgements The authors thank the Japanese women living in Bali who participated in this study. We would like to gratefully thank Dr. Indermohan S Narula, Team Leader, Local Fund Agent, Global Fund for TB, AIDS/HIV and Malaria, Ministry of Health, Mongolia for his assistance with the review and English editing of the paper. Consent for publication Not applicable. Funding This work was supported by JSPS KAKENHI Grant Number JP17K12364. The funding source did not have any role in the design, collection, analysis or interpretation of the data and in the writing of the manuscript. 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Journal of caring sciences 2017, 6(4):303-314. 16. Afaya A, Yakong VN, Afaya RA, Salia SM, Adatara P, Kuug AK, Nyande FK: A Qualitative Study on Women's Experiences of Intrapartum Nursing Care at Tamale Teaching Hospital (TTH), Ghana. Journal of caring sciences 2017, 6(4):303-314. 17. Mukamurigo J, Dencker A, Ntaganira J, Berg M: The meaning of a poor childbirth experience - A qualitative phenomenological study with women in Rwanda. PLoS One 2017, 12(12):e0189371. 18. Rai RK: Tracking women and children in a Continuum of Reproductive, Maternal, Newborn, and Child Healthcare (RMNCH) in India. Journal of epidemiology and global health 2014, 4(3):239-243. 19. Small R, Roth C, Raval M, Shafiei T, Korfker D, Heaman M, McCourt C, Gagnon A: Immigrant and non-immigrant women's experiences of maternity care: a systematic and comparative review of studies in five countries. BMC Pregnancy Childbirth 201 14:152. 20. Sandelowski M: What's in a name? Qualitative description revisited. Res Nurs Health 2010, 3 21. Campbell JL, Quincy C, Osserman J, Pedersen OK: Coding In-depth Semistructured Interviews:Problems of Unitization and Intercoder Reliability and Agreement. Sociological Methods & Research 2013, 42(3):294-320. 22. Donabedian A: The quality of care. How can it be assessed? Jama 1988, 260(12):1743-1748. Page 12/17 Page 12/17 23. Halldorsdottir S, Karlsdottir SI: The primacy of the good midwife in midwifery services: an evolving theory of professionalism in midwifery. Scandinavian journal of caring sciences 2011, 25(4):806-817. 24. Horiuchi S, Yaju Y, Kataoka Y, Grace Eto H, Matsumoto N: Development of an evidence-based domestic violence guideline: supporting perinatal women-centred care in Japan. Midwifery 2009, 25(1):72-78. 25. Tong A, Sainsbury P, Craig J: Consolidated criteria for reporting qualitative research (COREQ): a 32-item checklist for interviews and focus groups. References International journal for quality in health care : journal of the International Society for Quality in Health Care 2007, 19(6):349-357. 26. Nababan HY, Hasan M, Marthias T, Dhital R, Rahman A, Anwar I: Trends and inequities in use of maternal health care services in Indonesia, 1986-2012. International journal of women's health 2018, 10:11-24. 27. Hunter A, Devane D, Houghton C, Grealish A, Tully A, Smith V: Woman-centred care during pregnancy and birth in Ireland: thematic analysis of women's and clinicians' experiences. BMC Pregnancy Childbirth 2017, 17(1):322. , , g , , y , g p g y thematic analysis of women's and clinicians' experiences. BMC Pregnancy Childbirth 2017, 17(1):322. 28. Pope R, Graham L, Patel S: Woman-centred care. International journal of nursing studies 200 29. Graneheim UH, Lundman B: Qualitative content analysis in nursing research: concepts, procedures and measures to achieve trustworthiness. Nurse education today 2004, 24(2):105-112. Krippendorff K: Content analysis: An introduction to its methodology: Sage publications; 1980. 31. Cavanagh S: Content analysis: concepts, methods and applications. Nurse researcher 1997, 4(3):5-16. 32. Ezzy D: Qualitative Analysis: Practice and Innovation. London: Taylor & Francis Ltd; 2002. 33. Elo S, Kyngas H: The qualitative content analysis process. Journal of advanced nursing 200 33. Elo S, Kyngas H: The qualitative content analysis process. Journal of advanced nursing 2008, 62(1):107-115. 34. Twinn S: An exploratory study examining the influence of translation on the validity and reliability of qualitative data in nursing research. Journal of advanced nursing 1997, 26(2):418-423. 35. Bengtsson M: How to plan and perform a qualitative study using content analysis. NursingP 35. Bengtsson M: How to plan and perform a qualitative study using content analysis. NursingPlus Open 2016, 2:8-14. ow to plan and perform a qualitative study using content analysis. NursingPlus Open 2016, 2:8 36. Carter N, Bryant-Lukosius D, DiCenso A, Blythe J, Neville AJ: The use of triangulation in qualitative research. Oncology nursing forum 2014, 41(5):545-547. 37. Chen SW, Hutchinson AM, Nagle C, Bucknall TK: Women's decision-making processes and the influences on their mode of birth following a previous caesarean section in Taiwan: a qualitative study. BMC Pregnancy Childbirth 2018, 18(1):31. 38. Iida M, Horiuchi S, Porter SE, Pope R, Graham L, Patel S: The relationship between women-centred care and women's birth experiences: a comparison between birth centres, clinics, and hospitals in Japan. Midwifery 2012, 28. 39. References Broughton E, Achadi A, Latief K, Nandiaty F, Nurhaidah, Qomariyah SN, Rianty T, Wahyuni S, AP E: Indonesia Hospital Accreditation Process Impact Evaluation:Midline Report. In.: USAID; 2015. 40. Syahriana S, Mappaware NA, Hasibuan Y, Elizawarda E, Usman AN: Type of Delivery and Length of Stay in Hypertension Patients. Global Journal of Health Science 2018, 10(6):158. 41. Sakanashi K, Katsukawa Y, Usui M, Nabeta M, Ohga A, Nagai S: Preference regarding conditions for postpartum early discharge and support system: view of medical profession. Yokohama Journal of Nursing 2011, 4(1), 71-77. https://ci.nii.ac.jp/naid/110008146960/. 42. Takahashi Y, Tamakoshi K: Factors associated with early postpartum maternity blues and depression tendency among Japanese mothers with full-term healthy infants. Nagoya journal of medical science 2014, 76(1-2):129-138. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345732/pdf/2186-3326-76-0129.pdf 43. Takeuchi J, Sakagami Y, Perez RC: The Mother and Child Health Handbook in Japan as a Health Promotion Tool: An Overview of Its History, Contents, Use, Benefits, and Global Influence. Glob Pediatr Health 2016, 3:2333794X16649884. 44. Mori R, Yonemoto N, Noma H, Ochirbat T, Barber E, Soyolgerel G, Nakamura Y, Lkhagvasuren O: The Maternal and Child Health (MCH) handbook in Mongolia: a cluster-randomized, controlled trial. PLoS One 2015, 10(4):e0119772. 45. UNICEF : UNICEF Data: Monitoring the Situation of Children and Women Delivery care data Caesarian section [https://data.unicef.org/resources/resource-topic/maternal-health/] Accessed 3 January 2019. 46. Betran AP, Ye J, Moller AB, Zhang J, Gulmezoglu AM, Torloni MR: The Increasing Trend in Caesarean Section Rates: Global, Regional and National Estimates: 1990-2014. PLoS One 2016, 11(2):e0148343. Page 13/17 Page 13/17 47. Boerma T, Ronsmans C: Global epidemiology of use of and disparities in caesarean sections - Authors' reply. Lancet 2019, 394(10192):25. 48. World Health Organization, HRP: WHO Statement on Caesarean Section Rates 2015 [http://www.who.int/reproductivehealth/publications/maternal_perinatal_health/cs-statement/en/]. Accessed 3 January 2019. 48. World Health Organization, HRP: WHO Statement on Caesarean Section Rates 2015 [http://www.who.int/reproductivehealth/publications/maternal_perinatal_health/cs-statement/en/]. Accessed 3 January 2019. 49. Pristya TY, Mas’udah AF: Section Cesarean in Urban Areas Indonesia: How the Relation with Frequencies of Antenatal Care? KnE Life Sciences 2018:377–382-377–382. 50. Bohren MA, Hofmeyr GJ, Sakala C, Fukuzawa RK, Cuthbert A: Continuous support for women during childbirth. Cochrane Database Syst Rev 2017, 7(7):Cd003766. 51. Munk-Olsen T, Laursen TM, Mendelson T, Pedersen CB, Mors O, Mortensen PB: Risks and predictors of readmission for a mental disorder during the postpartum period. Arch Gen Psychiatry 2009, 66(2):189-195. 52. References Munk-Olsen T, Laursen TM, Meltzer-Brody S, Mortensen PB, Jones I: Psychiatric disorders with postpartum onset: possible early manifestations of bipolar affective disorders. Arch Gen Psychiatry 2012, 69(4):428-434. 53. Dekel S, Stuebe C, Dishy G: Childbirth Induced Posttraumatic Stress Syndrome: A Systematic Review of Prevalence and Risk Factors. Front Psychol 2017, 8:560. 54. Schmied V, Black E, Naidoo N, Dahlen HG, Liamputtong P: Migrant women's experiences, meanings and ways of dealing with postnatal depression: A meta-ethnographic study. PLoS One 2017, 12(3):e0172385. 55. Gamble J, Creedy D, Moyle W, Webster J, McAllister M, Dickson P: Effectiveness of a counseling intervention after a traumatic childbirth: a randomized controlled trial. Birth (Berkeley, Calif) 2005, 32(1):11-19. 56. Mercer RT: The nurse and maternal tasks of early postpartum. MCN The American journal of maternal child nursing 1981, 6(5):341-345. 57. Matsunaga, Y: Attitude of married couples toward husband’s presence at child birth: A questionnaire survey. The Japanese Association of Rural Medicine 2014, 62(5), 779-784. doi: 10.2185/jjrm.62.779. 57. Matsunaga, Y: Attitude of married couples toward husband’s presence at child birth: A questionnaire survey. The Japanese Association of Rural Medicine 2014, 62(5), 779-784. doi: 10.2185/jjrm.62.779. 58. Sonobe M, Usui M, Kawamura A, Hirose T: Relationship between satisfaction with childbirth mother-child interaction one month postpartum. J Child Health 2012, 53:210-218. 58. Sonobe M, Usui M, Kawamura A, Hirose T: Relationship between satisfaction with childbirth mother-child interaction one month postpartum. J Child Health 2012, 53:210-218. 59. Horiuchi S, Kataoka Y, Eto H, Oguro M, Mori T: The applicability of women-centered care: Two case studies of capacity-building for maternal health through international collaboration. Japan Journal of Nursing Science 2006, 3(2):143-150. 59. Horiuchi S, Kataoka Y, Eto H, Oguro M, Mori T: The applicability of women-centered care: Two case studies of capacity-building for maternal health through international collaboration. Japan Journal of Nursing Science 2006, 3(2):143-150. 60. Iida M: Developing and testing of an instrument to assess women-centered care during pregnancy. J Jpn Acad Midwif 2010, 24(2):284-293. 60. Iida M: Developing and testing of an instrument to assess women-centered care during pregnancy. J Jpn Acad Midwif 2010, 24(2):284-293. 61. Morgan L: Conceptualizing Woman-Centred Care in Midwifery. Canadian Journal of Midwifery Research and Practice 2015, 14(1):8-15. https://www.researchgate.net/publication/281746956 61. Morgan L: Conceptualizing Woman-Centred Care in Midwifery. Canadian Journal of Midwifery Research and Practice 2015, 14(1):8-15. https://www.researchgate.net/publication/281746956 62. References Yanti Y, Claramita M, Emilia O, Hakimi M: Students' understanding of "Women-Centred Care Philosophy" in midwifery care through Continuity of Care (CoC) learning model: a quasi-experimental study. BMC Nurs 2015, 14:22. 62. Yanti Y, Claramita M, Emilia O, Hakimi M: Students' understanding of "Women-Centred Care Philosophy" in midwifery care through Continuity of Care (CoC) learning model: a quasi-experimental study. BMC Nurs 2015, 14:22. 63. Tinkler A, Quinney D: Team midwifery: the influence of the midwife-woman relationship on women's experiences and perceptions of maternity care. Journal of advanced nursing 1998, 28(1):30-35. 63. Tinkler A, Quinney D: Team midwifery: the influence of the midwife-woman relationship on women's experiences and percept of maternity care. Journal of advanced nursing 1998, 28(1):30-35. 64. Fleissig A: Are women given enough information by staff during labour and delivery? Midwifery 1993, 9(2):70-75. 64. Fleissig A: Are women given enough information by staff during labour and delivery? Midwifery 1993, 9(2):70-75. Table Page 14/17 Figures Figures Page 15/17 Figure 1 Themes and categories of the childbirth experiences of Japanese women in Bali Description Note: Themes are in italics and categories are listed below the themes Figure 1 Themes and categories of the childbirth experiences of Japanese women in Bali Description Note: Themes are in italics and categories are listed below the themes Page 16/17 Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. Additionalfile1.docx Additionalfile1.docx Additionalfile2.docx Additionalfile2.docx Page 17/17 Page 17/17	33,944
ac172013fb1ea00311df2412a7dbcacc_2	French-Science-Pile	Open Science	Various open science	2,010	Learning in disrupted projects: on the nature of corporate and personal learning. International Journal of Production Research, 2007, 45 (12), pp.2775-2797. ⟨10.1080/00207540600774083⟩. ⟨hal-00512915⟩	None	English	Spoken	5,153	9,261	Jefferson Harvey Fo Basic LC 8.44% 7.01% Disaggregated LC 4.01% 5.30% Proportional difference 0.48 0.76 Cost of disruption over 100 products when splitting PL & CL ee rP Note that if Jefferson’s assertions were taken over 100 products (rather than 1000 items) there rR would be a lower CLCI and the disaggregated LC would be 6.48% (rather than 4.01%) giving a proportional difference of 0.77 (equal to that for Harvey). iew 6.3 Exploring the role of asymptotes ev One of the assertions used in the above example is that all labour hours can benefit from both PL and CL. If we now assume that this is not the case, and that, instead, PL is possible on On only a certain proportion of the task, then this means that there is likely to be a limit to the extent to which the time to undertake a task can be reduced. This leads on to consideration of ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research an asymptote for PL gains. The role of such an asymptote will be discussed next. When disaggregating the learning curve it seems plausible to consider the proportion of the time taken to complete a task that can benefit from either PL or CL. For example, a typical learning curve projection assumes that PL is possible on all of the hours involved in the 1st unit – that is the asymptote is zero. In addressing the implementation of the learning curve on 20 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research personal learning it may be important to establish the extent of learning that can be attained the position of the asymptote. The above sections have discussed the consideration of the learning rate and overall labour hours when disaggregating a LC between PL and CL. This was done within the context of the most common type of learning curve model - the log-linear model. A common amendment to Fo the log-linear model is to incorporate a plateauing effect (Conway and Schultz (1959); Baloff (1966)) or a non-zero asymptote (DeJong (1957)). rP An indication of the proportion of the task where PL is possible is therefore relevant to the ee disaggregation of the learning curve within the context of managing a disrupted project. This rR is related to the so-called “plateauing effect”. A plateauing effect reflects the situation where labour ceases to learn and is the equivalent of a non-zero asymptote. The most common ev reason for plateauing - a non-zero asymptote - to be incorporated into a LC is because there may be a limit to the minimum time taken to complete a task regardless of how many iew repetitions are made. DeJong (1957)) describes this as incompressibility. On When considering whether or not asymptotes do occur, Dar-El (2000)(2000, p19) notes that there are “two references in the literature which state that learning continues even in the ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Page 22 of 40 millions, however, in practice when the number of cycles runs into the thousands performance times tend to asymptote”. An asymptote is used in curves such as DeJong’s (DeJong (1957)) model, which adapts a loglinear model by incorporating an incompressibility factor. This is an allowance that is made because machine-time is not compressible as the number of units increases. Dar-El (2000, 21 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 23 of 40 p70) argues that “any review of learning curve literature, would refer to DeJong’s model as ‘appealing’ – but in reality, it was useless – because of our inability to find an accurate value for M [the incompressibility factor]”. However, Cochrane (1968) quoted in Teplitz (1991), supplies sample incompressibility factors. For example he suggests that the asymptote for a machine shop operation occurs at 77% of the hours taken to produce unit 1 whereas for aircraft subassembly the asymptote occurs at 43%. Fo When considering the existence of an asymptote for PL or CL, De Jong (DeJong (1957)) rP states, “where no technical or organizational changes are made for a given task [that is corporate learning], it appears – as is only to be expected – that [Tn] tends to approach a ee certain limit value”(p53, our emphasis). This implies that it is likely that an asymptote exists rR for PL. That is, the potential amount of PL to be gained is restricted by the given set of tasks that need to be performed. ev However, DeJong (DeJong (1957), p53) also states “in the … case…when technical iew equipment and organization are gradually improved [that is, corporate learning is included] it does repeatedly happen that the fall in [Tn] is in accordance with [the normal LC formula - On with no asymptote] even over a relatively long period”. DeJong is therefore suggesting that an asymptote does not occur for the overall LC that includes both PL and CL. The reason for ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research this could be that CL can potentially remove or replace tasks, change the nature of tasks through, for example, tooling, and reduce the size of the task to be completed. Thus, there is no obvious non-zero asymptote for CL. The basic task that is to be performed could dramatically change throughout the project due to CL. Thus, we consider the following situation for an overall curve: 22 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research If we take the situation of aircraft assembly, then Cochrane (1968) asserts that an overall asymptote should occur at 43% of the hours of the first unit. We intend to split the curve between PL & CL and it has been argued that PL may have an asymptote, but that CL does not. Based on Jefferson’s assertions, we use an overall LCI of 75%. The basic LC is split between Fo PL and CL with the value of CLCI and PLCI being calculated in order to minimise the variance between the basic LC and the split LC over 1000 products and the PL gain over rP 1000 products is approximately 22% of the overall learning gain. This results in a PLCI of 95.3% and a CLCI of 86.1%. rR ee For Harvey’s assertions, the same procedure results in a PLCI of 93.6% and a CLCI of 85.7%. The difference between the basic LC and the disaggregated curve with an asymptote is shown in graph 6 below. ev Graph 6: ABOUT HERE. iew As for the previous example, the two curves are not the same. However, as before, we use On them to compare the impact of the design change disruption. Graphs 7 and 8 demonstrate the impact of the design change on each of these curves. Graph 7: ABOUT HERE ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Graph 8: ABOUT HERE As for the previous example, the main observation from Graphs 7 and 8 is that the additional hours that are expected from the impact of the design change (that is, the area between the 23 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 24 of 40 Page 25 of 40 solid and dotted lines) is less when PL and CL are split compared to the impact on the basic LC. The cost of the disruption in each case is as follows: Jefferson Harvey Cost of disruption over 100 products when Fo splitting PL & CL with a non-zero asymptote Basic LC 8.44% 7.01% Disaggregated LC 4.06% 5.28% Proportional difference 0.48 0.75 rR ee rP In the case of this particular example disruption, the cost of the disruption is similar to the previous example where asymptotes was not taken account of, and the shapes of the graphs are not significantly different. iew ev 6.4 Inter-relationship between corporate and personal learning We have indicated above that gains in PL impact the extent of gains from CL, that is, On introducing CL late in a project is likely to cost more in wasted PL than if introduced early. Thus, CL also impacts the gains in PL. For example, if PL can be shared and routinized ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research across workers, then this will become CL. However, if the organisation makes improvements to its processes, then this will have an impact on individual workers learning. For example, if CL involves the introduction of new tools then individual workers will lose the learning that they have built up on the use of previous tools and so require to relearn the task. Also, if CL involves the introduction of new instructions on how to carry out a task, then this will affect the rate at which individual workers will learn. Therefore, PL must also be a function of CL. 24 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research It was argued earlier that there is only a proportion of all of the tasks, or of each task, which is amenable to PL. However, in this case the amount of PL that can be obtained is a continuous function of the CL applied. For Jefferson, a 75% learning curve over 1000 sets reduces the time taken on unit 1 to 5.7% Fo of its value by the 1000th set. If it were assumed that a PL log curve could be produced and that its initial value was taken as the total time taken on the 1st unit, then to meet Jefferson’s rP assertion, a PLCI of approximately 97.7% would be required. However, this assumes that there is no interaction between CL and PL. When PL and CL are considered to be inter- ee related, a more complex approach is required. rR As CL and PL are inter-related the curve will be constructed iteratively. For example, the ev amount of PLG (Personal Learning Gain) and CLG for unit 1 will be required to be calculated, these will then feed into the calculations for PLG and CLG for unit 2 and so on. iew Thus, the labour-hours required to unit 3 (T3) will be equal to: On T3 = T1 – PLG1 – CLG1 – PLG2 – CLG2 ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 And PLG2 = f(CLG1), CLG2 = f(PLG1) Where, in general: T1 = Labour-hours required to manufacture product 1 PLGi = Number of labour-hours learning gain between product i and product i+1 due to Personal Learning 25 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 26 of 40 Page 27 of 40 CLGi = Number of labour-hours learning gain between product i and product i+1 due to Corporate Learning Not only are CL and PL a function of each other, but an organisation may choose not to invest in CL because the extent of total PL gained over the production run does not warrant the investment. This would be particularly relevant for projects where the number of items to Fo be manufactured is not large. Therefore, investment in CL is a judgment made by an organisation dependant upon the extent of PL that has already been gained by workers. For rP this reason it becomes important to be able to compute the extent of total PL gained before the implementation of the proposed CL. Thus, disaggregation is important, not only when ee considering disruptions but also when considering new proposals for achieving CL. When a rR disruption occurs on a project, an iterative model can be used to help to assess the impact of a disruption on the expected learning gains for a project. The expected learning gains for both ev PL and CL, prior to the occurrence of any disruptions, can be determined using the above procedure only. iew In order to illustrate the impact of an iterative model the following approach is taken: On i. Begin with T1: This may be known because unit 1 has already been completed, ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research otherwise it is calculated by going back up the overall learning curve from a standard time at unit x. ii. Determine PL Curve: Assuming the asymptote is zero, then based on an estimate for the PLCI, calculate PLG1, PLG2, PLGi and so on. Based on Jefferson’s assertions, PLCI is 97.7% and for Harvey a PLCI of 96.8%. 26 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research iii. Calculate the CL gains between each product as the total gain between each product (based on the overall LC) minus the PL Gain between each product. This will result in a CL that is not a log curve. Based on this approach, the impact of the design change considered in the previous example would be as shown in graphs 9 and 10 below. rP Fo Graph 9: ABOUT HERE Graph 10: ABOUT HERE The cost of the disruption in each case is as follows: ee Jefferson Cost of disruption over 100 rR Harvey products when PL & CL Interact Basic LC Disaggregated LC 7.01% 5.18% 4.58% 0.61 0.65 iew Proportional difference 8.44% ev 8. Discussion ly On 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 The table below shows a summary of the results from the example disruption when considered from each of the four approaches. It is unsurprising that in each of the disaggregated cases the impact of the disruption is less than when calculated using the basic learning curve model. Given the nature of this particular example disruption, the differences are such that, from the perspective of labour planning, disaggregation is likely to lead to the more efficient use of labour. Nevertheless, for reasons identified earlier, using Jefferson as 27 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 28 of 40 Page 29 of 40 the basis for calculations might lead to too little labour being planned for and so possible further self-induced disruption from the need for accelerating actions. However, for the interactive model there is agreement. Thus, it seems likely that disaggregation would be useful when considering disruptions of the sort discussed at the beginning of this paper. At the extremes, the role of disaggregation is Fo obviously less significant: if all CL undertaken before the manufacturing commences (here there must be an expectation of no disruptions to the design of the product after production rP starts) then only PL can be gained during the project; if all of the tasks only involve machinery, then all CL and no PL to be gained during project. rR ee The explorations above do suggest that it is feasible to use the sparse assertions from the literature (Jefferson and Harvey) to undertake a plausible quantitative evaluation. However, ev the consequences of using each of these assertions may, in many instances, provide very different forecasts of labour requirements, except when an interaction between PL and CL is iew considered. Jefferson SUMMARY Harvey On Disruption of Basic LC 8.44% 7.01% Cost of disruption over 100 products when 4.01%/ 0.48 5.30%/ 0.76 4.06%/ 0.48 5.28%/ 0.75 Cost of disruption over 100 products when PL 5.18%/ 0.61 4.48%/ 0.65 ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research splitting PL & CL Cost of disruption over 100 products when splitting PL & CL with a non-zero asymptote & CL Interact 28 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research The explorations undertaken have demonstrated that the disaggregation of a learning curve with respect to disruptions is a non-trivial task. The paper has highlighted a number of issues with current theories when considering the disaggregation of a LC. The 5 key elements of disaggregation are: Fo (i) determining the boundary between PL and CL; (ii) determining whether either, or neither of PL and CL be represented as smooth log curves; rP (iii) establishing the typical learning rates for PL and CL and the proportion of overall learning split between the two; ee (iv) establishing whether PL and/or CL learning reach an asymptote. rR (v) although it seems plausible that PL and CL interact, an interactive model has rarely been explored, and if it were used then how are PL and CL related. ev In this paper we have been considering the relatively short run of 100 items. However in long iew run cases there may be a need to account for the so called “dual-phase model” (Dar-El et al. (1995)). This seeks to account for the different rate of learning for cognitive task as compared On to motor tasks, where cognitive learning is faster that motor learning. The LCI changes as the dominance of cognitive learning shifts to the dominance of motor learning. ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Through an exploration of the disaggregation process, a number of management insights have been gained. An understanding of the process of disaggregation forces managers to ask important questions that may not be addressed otherwise. • The analysis draws attention to the differential impacts of personal and corporate learning under circumstances of disruption – managers are better able to consider 29 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 30 of 40 Page 31 of 40 appropriate mitigation actions. Similarly, it is only when seeking to disaggregate that the question as to how many of the new tasks that may be required following disruption have been undertaken by the organization before becomes relevant and necessary. • The process highlights that disaggregation is important, not only when considering disruptions but also, when considering proposals for gaining CL at the potential cost Fo of wasted PL. • In addition, the requirement to make at least an assessment of the rate of personal rP learning may also have repercussions on non-disrupted learning estimates and so labour force targets. • ee The achievement of CL can change the nature of the tasks that are required to be rR performed and therefore suggest the requirement to consider the position of the PL asymptote, because the CL may change this. ev The test of the adequacy of any of the models derives from their ability to differentiate the iew consequences of different disruptions and so help in the ability to think through what to do as a manager, beyond any considerations for quantifying the real cost of learning losses from On disruptions. 33 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 34 of 40 Page 35 of 40 Basic LC & Disaggre gate d with asymptote and disruption - Je fferson Basic LC Disaggregated LC with asymptote 1200 Fo Labor Hours 1000 800 600 400 200 0 5 Basic LC with disruption 10 15 20 ee 0 Disaggreagted LC with disruption rP 25 30 P roduct rR Graph 7: Impact of disruption on basic and disaggregated (with asymptote) LC’s for Jefferson’s assertions iew ev Basic LC & Disaggregated w ith asymptote and disruptions Harvey Basic LC Disaggregated LC with asymptote 1000 Basic LC wth disruption Labor Hours 800 On Disaggregated LC with disruption 600 400 ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 International Journal of Production Research 200 0 0 5 10 15 20 25 30 Product Graph 8: Impact of disruption on basic and disaggregated (with asymptote) LC’s for Harvey’s assertions 34 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk International Journal of Production Research Basic LC & PL/CL Interact - Jefferson 1000 Basic LC & PL/CL Interac t LC Basic LC w ith dis ruption Interact PL/CL w ith disruption 600 400 200 0 5 10 15 20 ee 0 rP Labor Hours 800 Fo 25 30 Product Graph 9: Impact of disruption on basic and disaggregated (with interacting PL & CL) LC’s for Jefferson’s rR assertions ev Basic LC v PL/CL Interact - Harvey iew 1000 Basic LC & PL/CL Interact LC PL/CL interact w ith disruption Basic LC w ith disruption Labor Hours 800 On 600 400 200 ly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 0 0 5 10 15 20 25 30 Product Graph 10: Impact of disruption on basic and disaggregated (with interacting PL & CL) LC’s for Harvey’s assertions 35 http://mc.manuscriptcentral.com/tprs Email: ijpr@lboro.ac.uk Page 36 of 40 Page 37 of 40 Notes 1. 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https://openalex.org/W1553711420	OpenAlex	Open Science	CC-By	2,006	Prevalence of metabolic syndrome-related disorders in a large adult population in Turkey	S. Yavuz Sanisoğlu	English	Spoken	5,745	10,298	BioMed Central BioMed Central BioMed Central Open Acc Research article Prevalence of metabolic syndrome-related disorders in a large adult population in Turkey S Yavuz Sanisoglu1, Cagatay Oktenli2, Adnan Hasimi3, Mehmet Yokusoglu4 and Mehmet Ugurlu5 Open Access Address: 1Department of Biostatistics, Gülhane Military Medical Academy, Ankara, Turkey, 2Department of Internal Medicine, Gülhane Military Medical Academy, Ankara, Turkey, 3Department of Biochemistry and Clinical Biochemistry, Gülhane Military Medical Academy, Ankara, Turkey, 4Department of Cardiology, Gülhane Military Medical Academy, Ankara, Turkey and 5Turkish Ministry of Health, Ankara, Turkey Email: S Yavuz Sanisoglu - biyoistatistik@gmail.com; Cagatay Oktenli - coktenli@gata.edu.tr; Adnan Hasimi - ahasim@gata.edu.tr; Mehmet Yokusoglu - myokusoglu@gata.edu.tr; Mehmet Ugurlu - drmehmetugurlu@yahoo.com * Corresponding author * Corresponding author Received: 18 October 2005 Accepted: 10 April 2006 Published: 10 April 2006 Received: 18 October 2005 Accepted: 10 April 2006 BMC Public Health2006, 6:92 doi:10.1186/1471-2458-6-92 This article is available from: http://www.biomedcentral.com/1471-2458/6/92 This article is available from: http://www.biomedcentral.com/1 © 2006Sanisoglu et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Main Geographical Regions and Location of the Study Centers. syndrome to be 21.8% among adults in the United States [9-11]. Background Turkey, with its large land area, more than 70 million peo- ple, and growing economy, resembles a bridge in geo- graphical aspects between Europe and Asia. There are few existing large population studies on the epidemiology of metabolic syndrome-related disorders of Turkey. Turkish Adults Risk Factor Study (TEKHARF) was the first study, representing the whole country, conducted by the Turkish Cardiology Association [1]. The purpose of this study was to assess the prevalence of the metabolic syndrome-related disorders in the Turkish adult population of more than 15,000 persons, to address sex, age, educational and geographical differences, and to examine blood pressure, BMI, fasting blood glucose and serum lipids in Turkey. The etiology, prevention, and treatment of the metabolic syndrome currently are the focus of the intense research activities. The combination of abdominal obesity, hyper- tension, dyslipidemia, hyperglycemia, and insulin resist- ance or type 2 diabetes mellitus defines the metabolic syndrome [2]. The metabolic syndrome is said to consist of a cluster of metabolic risk factors, including dyslipi- demia, impaired carbohydrate metabolism, obesity, and high blood pressure [3,4]. In 1999, the WHO published the first official definition of the metabolic syndrome [5]. The new IDF definition of the metabolic syndrome addresses both clinical and research needs and provides an accessible, diagnostic tool that is suitable for use in populations worldwide [6]. The pathogenesis of meta- bolic syndrome is still unclear, although some environ- mental factors, coupled with still largely unknown genetic factors, clearly interact to produce the syndrome [7,8]. Previous reports using population-based data from the Third National Health and Nutrition Examination Survey have estimated the age-adjusted prevalence of metabolic Abstract Background: There are few existing large population studies on the epidemiology of metabolic syndrome-related disorders of Turkey. The purpose of this study was to assess the prevalence of metabolic syndrome-related disorders in the Turkish adult population, to address sex, age, educational and geographical differences, and to examine blood pressure, body mass index, fasting blood glucose and serum lipids in Turkey. Methods: This study was executed under the population study "The Healthy Nutrition for Healthy Heart Study" conducted between December 2000 and December 2002 by the Health Ministry of Turkey. Overall, 15,468 Caucasian inhabitants aged over 30 were recruited in 14 centers in the seven main different regions of Turkey. The data were analyzed with the Students' t, ANOVA or Chi-Square tests. Results: Overall, more than one-third (35.08 %) of the participants was obese. The hypertensive people ratio in the population was 13.66 %, while these ratios for DM and metabolic syndrome were 4.16 % and 17.91 %, respectively. The prevalence of hypertension, metabolic syndrome and obesity were higher in females than males, whereas diabetes mellitus was higher in males than females. The prevalence of metabolic syndrome and related disorders were found to be significantly different across educational attainments for both men and women. The prevalence of hypertension increased with age, while it was remarkable that in the age group of 60–69 years, prevalence of diabetes mellitus and metabolic syndrome reached a peak value and than decreased. For obesity, the peak prevalence occurred in the 50–59 year old group. The prevalence of metabolic syndrome and related disorders were found to be significantly different according to geographical region. Conclusion: In conclusion, high prevalence of obesity and metabolic syndrome, particularly among women, is one of the major public health problems in Turkey. Interestingly, obesity prevalence is relatively high, but the prevalence of hypertension and hypercholesterolemia is relatively low in Turkish people. Future studies may focus on elucidating the reasons behind this controversy. Our findings may be helpful in formulating public health policy and prevention strategies on future health in Turkey. Page 1 of 6 (page number not for citation purposes) (page number not for citation purposes) BMC Public Health 2006, 6:92 http://www.biomedcentral.com/1471-2458/6/92 Main Geographical Regions and Location of the Study Centers Figure 1 Main Geographical Regions and Location of the Study Centers. Main Geo Figure 1 g p g y g Main Geographical Regions and Location of the Study Centers. Page 2 of 6 (page number not for citation purposes) Results A total of 2096 (13.6 %) were from Marmara, 1555 (10.1 %) were from Eastern Anatolia, 2541 (16.4 %) were from Southeast Anatolia, 2554 (16.5 %) were from Mediterra- nean, 2519 (16.3%) were from Aegean, 2190 (14.2 %) were from Black Sea, and 2013 (13.0 %) were from Cen- tral Anatolia. Based on self-report, 20.3% were illiterate (n = 3123), 18.0 % were literate only (n = 2766), 41.2% of the people (n = 6334) were elementary education, 13.1 % were high school graduate (n = 2015), and 7.4% were university educated (n = 1136). Study participants were studied in six separate age groups, namely those aged 30–39, 40–49, 50–59, 60–69, 70–79 and those aged older than 80. The survey was representa- tively stratified for sex, age, and for rural-urban distribu- tion. Overall, more than one-third (35.08 %) of the partici- pants was obese. The hypertensive people ratio in the pop- ulation was 13.66 %, while these ratios for DM and metabolic syndrome were 4.16 % and 27.38 %, respec- tively. Seven measures representing the metabolic syndrome were obtained, including fasting blood glucose, BMI, HDL-C and Low-density lipoprotein cholesterol (LDL-C), triglycerides, systolic blood pressure, and diastolic blood pressure. At the baseline examination, blood samples were taken after a minimum 6-hour fast. Serum was sepa- rated on-site within 30 minutes of venipuncture, stored at -4°C, and analyzed within 24 hours of venipuncture. Determination of routine biochemical parameters was performed with standard techniques by using an autoan- alyser. Values for each person were calculated by Friede- wald's formula and LDL-C values >10.36 mmol/l were not taken into account. The prevalence of hypertension, metabolic syndrome and obesity were higher in females than males, whereas diabe- tes mellitus was higher in males than females (Table 1). There were no statistically significant difference between the urban and rural areas with respect to metabolic syn- drome and related disorders such as hypertension, diabe- tes mellitus, and obesity (data not shown). The prevalence of metabolic syndrome and related disorders were found to be significantly different across educational attain- ments for both men and women (Table 1). Obese men and women tended to have lower educational attainment (Table 1). The prevalence of hypertension increased with age, while it was remarkable that in the age group of 60– 69 years, prevalence of diabetes mellitus and metabolic syndrome reached a peak value and than decreased (Table 1). Methods This study was executed under the population study "The Healthy Nutrition for Healthy Heart Study" conducted between December 2000 and December 2002 by the Health Ministry of Turkey. Overall, 15,468 Caucasian inhabitants aged over 30 were recruited in 14 centers in the seven main different regions of Turkey that would rep- resent whole Turkish People in terms of living conditions and geographic background (Figure 1). Marmara, Aegean and Mediterranean regions are in the western part of Tur- key, while Eastern and South-Eastern Anatolia are in the eastern part. Black Sea region is in the northern side. The number of people for each center was calculated by using stratified sampling technique. In the selection of the par- ticipants, we used random number table. In order to cover the whole population of Turkey, men and women were randomly selected from urban and rural areas from all regions. Sample size for the study was calculated by allow- ing for 0.25% error in prevalence with 95% confidence interval (CI). At the end of the study, we reached 15,468 Page 2 of 6 (page number not for citation purposes) Page 2 of 6 (page number not for citation purposes) BMC Public Health 2006, 6:92 http://www.biomedcentral.com/1471-2458/6/92 http://www.biomedcentral.com/1471-2458/6/92 people. The response rate was 87.3 %. Sample sizes for each health center were calculated using the census of the year 2000 by stratified sampling method [12]. Data form was developed after the execution of a pilot study. All socio-demographic data (age, gender, and educational level), and medical history were obtained through a med- ical staff by using a standardized questionnaire in the study centers. Educational attainment was categorized as illiterate, literate only, elementary education, high school graduate or a university education. Completion of the questionnaire was considered to imply informed consent. The study was carried out in accordance with the guide- lines of the Helsinki Declaration of Human Studies. used to compare the parameters of more than two groups. Relations among the categorical parameters were investi- gated by "Chi-square test". pvalues less than or equal to 0.05 were considered as statistically significant. Page 3 of 6 (page number not for citation purposes) Discussion This study is one of the largest population-based studies of metabolic syndrome-related disorders ever conducted in Turkey. The prevalence of metabolic syndrome was 10.09 and 27.33 % in men and women, respectively. Onat et al. reported a higher prevalence of the metabolic syn- drome in Turkish adults (27% in men and 38.6% in women) than our findings [13]. In the National Health and Nutrition Examination Survey sample in U.S. adults [11], age-adjusted prevalence of the metabolic syndrome was 24.0% and 23.4% in men and women, respectively. In the current study, the prevalence of diabetes was deter- mined to be 4.16% (5.22% in men and 3.69% in women). Onat et al. reported that prevalence of diabetes in a Turkish population was 3.4% (4.2% for men and 2.8% for women) [13]. In The Turkish Diabetes Epidemi- ology Study (TURDEP), the prevalence of diabetes was 7.4% (6.2% in men and 8.0% in women) [14]. On the other hand, the overall prevalence of obesity was 35.08 % (21.16% in men and 41.32% in women) in our study. Similar to our results, in a large multicenter nationwide study [1], this prevalence was found to be 25.3% for Turk- ish males and 44.2% for Turkish females. Conversely, Sat- man et al. reported that the low prevalence of obesity in adults in Turkey was 22.3% (12.9% in men, 29.9% in women) [14]. Finally, the prevalence of hypertension in our study (13.66%) was relatively lower than a previous study (46%) [15]. We found that the prevalence of diabetes among the regions was the highest in South-East Anatolia region. A study conducted by Gokcel et al. [20] in Adana, a city in South-East Anatolia, reported the diabetes prevalence as 11.6%. On the contrary, as reported previously [33], the lowest prevalence of diabetes was obtained from the par- ticipants living in the Eastern Anatolia Region of Turkey. In consistent with our observation, in Central Anatolia, the prevalence of diabetes was reported as 6.4% [19] and 6.9% [34]. Similar to our results, the prevalence of diabe- tes was 5.2% in Trabzon, a city located in the Black Sea Region. On the other hand, in contrast to a previous report [14], subjects living in the Eastern and South-East Anatolia Region of Turkey had the highest prevalence of obesity in our study. However, consistent with the same report [14], obesity prevalence in Central Anatolia was high. triglyceride in the 70–79 age group, and triglyceride in 40–49 age group were significantly different (Table 3). over 50 years met the criteria for the metabolic syndrome [11]. In this context, aging may be a risk factor for meta- bolic syndrome. In many studies, it was also reported that the prevalence of diabetes increased with age [16-20]. Likewise, in the current study, the prevalence of obesity increased markedly from the 30- to 39-year-old age group to the 50- to 59-year-old age group and then decreased. In many studies, it has been reported that prevalence of obesity increases with age [21-23]. This can be explained partly by a decrease in the degree of physical activity by aging [21,24]. over 50 years met the criteria for the metabolic syndrome [11]. In this context, aging may be a risk factor for meta- bolic syndrome. In many studies, it was also reported that the prevalence of diabetes increased with age [16-20]. Likewise, in the current study, the prevalence of obesity increased markedly from the 30- to 39-year-old age group to the 50- to 59-year-old age group and then decreased. In many studies, it has been reported that prevalence of obesity increases with age [21-23]. This can be explained partly by a decrease in the degree of physical activity by aging [21,24]. Mean total cholesterol level of our study population was 4.92 mmol/l (4.91 mmol/l in men and 4.92 mmol/l in women). Hypercholesterolemic levels were recorded in 29.89% in men, 28.35 in women in the present study. Mean LDL-C level was 2.92 mmol/l (2.96 mmol/l in men, 2.90 mmol/l in women). High values of LDL-C were found 29.89% in men and 28.35% in women in our study. Mean concentrations of HDL-C, were 1.10 mmol/l (1.10 mmol/l in men and women). In the current study, lower HDL-C concentrations were found 25.16% in men and 32.39% in women. In our study, mean concentra- tions of triglyceride were 1.76 mmol/l (1.80 mmol/l in men and 1.75 mmol/l in women). In addition, prevalence of hypertriglyceridemia in our study was 35.54% (40.72% in men and 33.23% in women). The prevalence of diabetes, obesity, hypertension and metabolic syndrome was found to be decreased with high educational levels in our study. The results are in line with studies conducted in Turkey and Europe [21,25-27,31]. In a recent study [25], a clear association was reported between education and blood glucose levels and a higher risk of diabetes associated with lower levels of education. triglyceride in the 70–79 age group, and triglyceride in 40–49 age group were significantly different (Table 3). It is known that unhealthy dietary habits as a high fat intake and low fruit and vegetable intake, as well as phys- ical inactivity, are inversely related with educational level [32]. Page 4 of 6 (page number not for citation purposes) Results For obesity, the peak prevalence occurred in the 50–59 year old group. The prevalence of metabolic syndrome and related disorders were found to be significantly differ- ent according to geographical region (Table 1). Because waist circumference was not determined in this study, we defined the metabolic syndrome using BMI (≥ 30 kg/m2 for both men and women) instead of waist cir- cumference. BMI was calculated as the ratio of body weight to square of body height (kg/m2). Obesity was defined based on BMI 30 kg/m2. Blood pressure was cal- culated as the average of three measurements taken under standardized conditions in a supine position with a sphygmomanometer. Diabetes mellitus was defined as a fasting blood glucose level = 7 mmol/l, or reported use of diabetes medication. The IDF definition was used for the diagnosis of the metabolic syndrome [6]. The distribution across regions of biochemical parameters by geographical region, gender and age groups is pre- sented in the Table 2 and 3. The differences of biochemi- cal parameters among the geographical regions were significantly different (Table 2), while they, except HDL- C, were significantly different among the age groups (Table 3). When the data compared according to gender, we found that total cholesterol and LDL-C in the age groups of 30–39, 50–59 and 60–69, total cholesterol and A total of data were analyzed by StatsDirect (Ver 2.2.0, Stats Direct Ltd, UK) and SPSS 11.5 (SPSS Inc., Chicago, Il., USA) software. Descriptives were shown either the per- centages or mean ± SD for categorical and continuous var- iables, respectively. In order to minimize the effects of the outliers we calculated the 5% trimmed means. Parameters of two group of people were compared by "Independent samples t test". "Analysis of variance (ANOVA test)" was Page 3 of 6 (page number not for citation purposes) Page 3 of 6 (page number not for citation purposes) http://www.biomedcentral.com/1471-2458/6/92 http://www.biomedcentral.com/1471-2458/6/92 http://www.biomedcentral.com/1471-2458/6/92 http://www.biomedcentral.com/1471-2458/6/92 BMC Public Health 2006, 6:92 triglyceride in the 70–79 age group, and triglyceride in 40–49 age group were significantly different (Table 3). Competing interests The author(s) declare that they have no competing inter- ests. 11. Ford ES, Giles WH, Dietz WH: Prevalence of the metabolic syn- drome among US adults: Findings from the third National Health and Nutrition Examination Survey. JAMA 2002, 287:356-359. http://www.biomedcentral.com/1471-2458/6/92 Onat A, Ceyhan K, Erer B, Basar O, Uysal O, Sansoy V: Systolic, diastolic pressure and pulse pressure as coronary risk factors in a population with low cholesterol levels Prospective 10- year evaluation. Clin Cardiol 2003, 26:91-97. 16. Ramachadran A, Snehalatha C, Satyavani K, Vijay V: Impaired fast- ing glucose and impaired glucose tolerance in urban popula- tion in India. Diabet Med 2003, 20:220-224. 17. Baltazar JC, Ancheta CA, Aban IB, Fernando RE, Baquilod MM: Prev- alence and correlates of diabetes mellitus and impaired glu- cose tolerance among adults in Luzon, Philippines. Diabetes Res Clin Pract 2004, 64:107-115. 18. Jaber LA, Brown MB, Hammad A, Nowak SN, Zhu Q, Ghafoor A, Herman WH: Epidemiology of diabetes among Arab Ameri- cans. Diabetes Care 2003, 26:308-313. 19. Ozdemir L, Topcu S, Nadir I, Nur N, Arslan S, Sumer H: The Prev- alence of Diabetes and Impaired Glucose Tolerance in Sivas, Central Anatolia, Turkey. Diabetes Care 2005, 28:795-798. 20. Gokcel A, Ozsahin AK, Sezgin N, Karakose H, Ertorer ME, Akbaba M, Baklaci N, Sengul A, Guvener N: High prevalence of diabetes in Adana, a southern province of Turkey. Diabetes Care 2003, 26:3031-3034 was lower in the western region (Mediterranean and Aegean Regions) than the northern region (Black Sea Region). Environmental factors or individual exposures such as salt intake may cause such differences in preva- lence among these populations. Table 1 – Prevalence of Metabolic Syndrome and Related Disorders Table 1 – Prevalence of Metabolic Syndrome and Related Disorders p gy y 15. Onat A, Ceyhan K, Erer B, Basar O, Uysal O, Sansoy V: Systolic, diastolic pressure and pulse pressure as coronary risk factors in a population with low cholesterol levels Prospective 10- year evaluation. Clin Cardiol 2003, 26:91-97. Table 2 – Distribution and Comparisons Results of the Biochemical Parameters for The Geographical Regions (5% Trimmed Mean) FBG, fasting blood glucose; TC, total cholesterol; TG, triglyceride y 16. Ramachadran A, Snehalatha C, Satyavani K, Vijay V: Impaired fast- ing glucose and impaired glucose tolerance in urban popula- tion in India. Diabet Med 2003, 20:220-224. 17. Baltazar JC, Ancheta CA, Aban IB, Fernando RE, Baquilod MM: Prev- alence and correlates of diabetes mellitus and impaired glu- cose tolerance among adults in Luzon, Philippines. Diabetes Res Clin Pract 2004, 64:107-115. Table 3 – Descriptive Statistics of the Parameters Accord- ing to the Age Groups and Gender (5% Trimmed Mean) F, female; M, male; FBG, fasting blood glucose; TC, total cholesterol; TG, triglyceride Table 3 – Descriptive Statistics of the Parameters Accord- ing to the Age Groups and Gender (5% Trimmed Mean) F, female; M, male; FBG, fasting blood glucose; TC, total cholesterol; TG, triglyceride 18. Jaber LA, Brown MB, Hammad A, Nowak SN, Zhu Q, Ghafoor A, Herman WH: Epidemiology of diabetes among Arab Ameri- cans. Diabetes Care 2003, 26:308-313. 19. Ozdemir L, Topcu S, Nadir I, Nur N, Arslan S, Sumer H: The Prev- alence of Diabetes and Impaired Glucose Tolerance in Sivas, Central Anatolia, Turkey. Diabetes Care 2005, 28:795-798. Conclusion In conclusion, high prevalence of obesity and metabolic syndrome, particularly among women, is one of the major public health problems in Turkey. Interestingly, obesity prevalence is relatively high, but the prevalence of hyper- tension and hypercholesterolemia is relatively low in Turkish people. Future studies may focus on elucidating the reasons behind this controversy. Our findings may be helpful in formulating public health policy and preven- tion strategies on future health in Turkey. g 6. Zimmet P, Alberti G, Shaw J: A new IDFworldwide definition of the metabolic syndrome: the rationale and the results. Dia- betesVoice 2005, 50:31-33 [http://www.diabetesvoice.org/issues/ 2005-09/issue_2005-09.pdf]. (accessed January 4, 2006) p ] ( J y ) 7. Liese AD, Mayer-Davis EJ, Haffner SM: Development of the mul- tiple metabolic syndrome: an epidemiologic perspective. Epidemiol Rev 1998, 20:157-172. 8. 9. 8. p 8. Bouchard C: Genetics and the metabolic syndrome. Int J Obes Relat Metab Disord 1995, 19(suppl 1):S52-59. 9. National Cholesterol Education Program: Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evalua- tion, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) 2002. ) 10. Park YW, Zhu S, Palaniappan L, Heshka S, Carnethon MR, Heymsfield SB: The metabolic syndrome: Prevalence and associated risk factors in the US population from the Third National Health and Nutrition Examination Survey, 1998–1994. Arch Intern Med 2003, 163:427-436. References 1. Onat A, Sansoy V, Soydan I, Tokgozoglu L, Adalet K: Türk Eriskin- lerde Kalp Sagligi, Risk Profili ve Kalp Sagligi. Ohan, Istanbul 2000. 2. Eckel RH, Grundy SM, Zimmet PZ: The metabolic syndrome. Lancet 2005, 365:1415-1428. 3. Timar O, Sestier F, Levy E: Metabolic syndrome X: a review. Can J Cardiol 2000, 16:779-789. In the current study, as in others [40,41], mean total cho- lesterol, LDL-C and HDL-C level of Turkish adults was lower than the Northern Europe and the Mediterranean populations [42]. The mean concentrations of triglyceride were higher than the Turkish Heart Study [43]. 4. Lindberg O, Tilvis RS, Strandberg TE, Valvanne J, Sairanen S, Ehnholm C, Tuomilehto J: Impacts of components of the metabolic syn- drome on health status and survival in an aged population. Eur J Epidemiol 1997, 13:429-434. 5. J p 5. World Health Organization: Definition, diagnosis and classifica- tion of diabetes mellitus and its complications: report of a WHO Consultation. Part 1: diagnosis and classification of diabetes mellitus. 1999 [http://whqlibdoc.who.int/hq/1999/ WHO_NCD_NCS_99.2.pdf]. Geneva, Switzerland: World Health Organization Discussion In the Black Sea Region, prevalence of obesity was found to be higher (36.89%) than a previous study (19.2%) [35]. The average values of total cholesterol, LDL- C, and triglyceride were also higher in South-Eastern Ana- tolia, Eastern Anatolia and Central Anatolia than the other regions. Consumption of foods with high fat and sugar content is also frequent in this area [36]. In our study, the prevalence of obesity in Marmara Region is consistent with a previous report [37] and this region has the lowest obesity prevalence as well as Aegean region. This may be due to participants from these regions consume more veg- etables, less carbohydrate; prefer olive oil or corn oil more when cooking [38,39]. The hypertension prevalence among the geographical areas in Turkey is also considera- bly different. Like a previous report [14], the prevalence In the present study, there was a clear age-related increase in the prevalence of the metabolic syndrome in Turkish adult population. The prevalence of metabolic syndrome was the lowest at age group 30–39 (15.34%), while it pro- gressively increased with age until the age group 50–59 (27.98%). Ford et al. reported the prevalence of metabolic syndrome increased with age, and 33–45% of subjects Page 4 of 6 (page number not for citation purposes) Page 4 of 6 (page number not for citation purposes) http://www.biomedcentral.com/1471-2458/6/92 Authors' contributions SYS carried out the data organization and performed the statistical analysis. CO participated in the evaluation of data. AH and MY contributed to the study with their knowledge on biochemistry and cardiology, respectively. MU participated in coordination and evaluation of data. 12. [http://www.die.gov.tr/nufus_sayimi/2000Nufus_Kesin1.htm]. 13 O A C h K B O E B T k S S V M t b [http://www.die.gov.tr/nufus_sayimi/2000Nufus_Kesin1.htm O A C O S S 13. Onat A, Ceyhan K, Basar O, Erer B, Toprak S, Sansoy V: Metabolic syndrome Major impact on coronary risk in a population with low cholesterol levels – A prospective and cross-sec- tional evaluation. Atherosclerosis 2002, 165:285-292. 14. Satman I, Yilmaz T, Sengul A, Salman S, Salman F, Uygur S, Bastar I, Tutuncu Y, Sargin M, Dinccag N, Karsidag K, Kalaca S, Ozcan C, King H, The TURDEP Group: Population-based study of diabetes and risk characteristics in Turkey: results of the Turkish Diabe- tes Epidemiology Study. Diabetes Care 2002, 25:1551-1556. Note Note Table 1 – Prevalence of Metabolic Syndrome and Related Disorders Table 2 – Distribution and Comparisons Results of the Biochemical Parameters for The Geographical Regions (5% Trimmed Mean) FBG, fasting blood glucose; TC, total cholesterol; TG, triglyceride Table 3 – Descriptive Statistics of the Parameters Accord- ing to the Age Groups and Gender (5% Trimmed Mean) F, female; M, male; FBG, fasting blood glucose; TC, total cholesterol; TG, triglyceride Acknowledgements This study was funded, organized and conducted by the Turkish Ministry of Health. The authors express their special thanks to the staff of 14 health screening centers. http://www.biomedcentral.com/1471-2458/6/92 BMC Public Health 2006, 6:92 References 1. Onat A, Sansoy V, Soydan I, Tokgozoglu L, Adalet K: Türk Eriskin- lerde Kalp Sagligi, Risk Profili ve Kalp Sagligi. Ohan, Istanbul 2000. 2. Eckel RH, Grundy SM, Zimmet PZ: The metabolic syndrome. Lancet 2005, 365:1415-1428. 3. Timar O, Sestier F, Levy E: Metabolic syndrome X: a review. Can J Cardiol 2000, 16:779-789. 4. Lindberg O, Tilvis RS, Strandberg TE, Valvanne J, Sairanen S, Ehnholm C, Tuomilehto J: Impacts of components of the metabolic syn- drome on health status and survival in an aged population. Eur J Epidemiol 1997, 13:429-434. 5. World Health Organization: Definition, diagnosis and classifica- tion of diabetes mellitus and its complications: report of a WHO Consultation. Part 1: diagnosis and classification of diabetes mellitus. 1999 [http://whqlibdoc.who.int/hq/1999/ WHO_NCD_NCS_99.2.pdf]. Geneva, Switzerland: World Health Organization 6. Zimmet P, Alberti G, Shaw J: A new IDFworldwide definition of the metabolic syndrome: the rationale and the results. Dia- betesVoice 2005, 50:31-33 [http://www.diabetesvoice.org/issues/ 2005-09/issue_2005-09.pdf]. (accessed January 4, 2006) 7. Liese AD, Mayer-Davis EJ, Haffner SM: Development of the mul- tiple metabolic syndrome: an epidemiologic perspective. Epidemiol Rev 1998, 20:157-172. 8. Bouchard C: Genetics and the metabolic syndrome. Int J Obes Relat Metab Disord 1995, 19(suppl 1):S52-59. 9. National Cholesterol Education Program: Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evalua- tion, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) 2002. 10. Park YW, Zhu S, Palaniappan L, Heshka S, Carnethon MR, Heymsfield SB: The metabolic syndrome: Prevalence and associated risk factors in the US population from the Third National Health and Nutrition Examination Survey, 1998–1994. Arch Intern Med 2003, 163:427-436. 11. Ford ES, Giles WH, Dietz WH: Prevalence of the metabolic syn- drome among US adults: Findings from the third National Health and Nutrition Examination Survey. JAMA 2002, 287:356-359. 12. [http://www.die.gov.tr/nufus_sayimi/2000Nufus_Kesin1.htm]. 13. Onat A, Ceyhan K, Basar O, Erer B, Toprak S, Sansoy V: Metabolic syndrome Major impact on coronary risk in a population with low cholesterol levels – A prospective and cross-sec- tional evaluation. Atherosclerosis 2002, 165:285-292. 14. Satman I, Yilmaz T, Sengul A, Salman S, Salman F, Uygur S, Bastar I, Tutuncu Y, Sargin M, Dinccag N, Karsidag K, Kalaca S, Ozcan C, King H, The TURDEP Group: Population-based study of diabetes and risk characteristics in Turkey: results of the Turkish Diabe- tes Epidemiology Study. Diabetes Care 2002, 25:1551-1556. 15. Pre-publication history The pre-publication history for this paper can be accessed here: 24. Gutierrez-Fisac JL, Regidor E, Rodriguez C: Economic and social factors associated with body mass index and obesity in the Spanish population aged 20–64 y. Eur J Public Health 1995, 5:193-198. http://www.biomedcentral.com/1471-2458/6/92/prepub 25. Stelmach W, Kaczmarczyk-Chalas K, Bielecki W, Stelmach I, Drygas W: How income and education contribute to risk factors for cardiovascular disease in the elderly in a former Communist country. Public Health 2004, 118:439-449. 26. WHO: Obesity: Preventing and Managing the Global Epidemic WHO Geneva, Switzerland; 1998. 27. Erem C, Arslan C, Hacihasanoglu A, Deger O, Topbas M, Ukinc K, Ersoz HO, Telatar M: Prevalence of Obesity and Associated Risk Factors in a Turkish Population (Trabzon City, Turkey). Obes Res 2004, 12:1117-1127. 28. Stam-Moraga MC, Kolanowski J, Dramaix M, Henauw SD, De Bac- quer DG, Kornitzer MD: Trends in the prevalence of obesity among Belgian men at work, 1977–1992. Int J Obes 1998, 22:988-992. 29. Quiles Izquierdo J, Vioque J: Prevalencia de obesidad en la Comunidad Valenciana. Med Clin 1996, 106:529-533. 30. Paeratakul S, Lovejoy JC, Ryan DH, Bray GA: The relation of gen- der, race and socioeconomic status to obesity and obesity co-morbidities in a sample of US adults. Int J Obes Relat Metab Disord 2002, 26:1205-1210. 31. Rosmond R, Bjorntorp P: Psychosocial and socio-economic fac- tors in women and their relationship to obesity and regional body fat distribution. Int J Obes Relat Metab Disord 1999, 23:138-145. 32. La Vecchia C, Negri E, Franceschi S, Parazzini F, Decarli A: Differ- ences in dietary intake with smoking, alcohol, and education. Nutr Cancer 1992, 17:297-304. 33. Onat A: Risk factors and cardiovascular disease in Turkey. Atherosclerosis 2001, 156:1-10. 34. Kelestimur F, Cetin M, Pasaoglu H, Coksevim B, Cetinkaya F, Unluhisarcý K, Unal S, Koker AH: The prevalence of identifica- tion of risk factors for type 2 diabetes mellitus and impaired glucose tolerance in Kayseri, Central Anatolia, Turkey. Acta Diabetol 1999, 36:85-91. 35. Erem C, Yildiz R, Kavgaci H, Karahan C, Deger O, Can G, Telatar M: Prevalence of diabetes, obesity and hypertension in a Turk- ish population (Trabzon city). Diabetes Res Clin Prac 2001, 54:203-208. 36. The State Institute of Statistics Prime Ministry Republic of Turkey: Household Consumption Expenditures Survey Results: 1994. Ankara, State Institute of Statistics Pub. Co.; 1997. 37. http://www.biomedcentral.com/1471-2458/6/92 http://www.biomedcentral.com/1471-2458/6/92 43. Mahley RW, Palaoglu KE, Atak Z, Dawson-Pepin J, Langlois AM, Che- ung V, Onat H, Fulks P, Mahley LL, Vakar F, Ozbayrakci S, Gokdemir O, Winkler W: Turkish Heart Study: lipids, lipoproteins, and apolipoproteins. J Lipid Res 1995, 36:839-859. 43. Mahley RW, Palaoglu KE, Atak Z, Dawson-Pepin J, Langlois AM, Che- ung V, Onat H, Fulks P, Mahley LL, Vakar F, Ozbayrakci S, Gokdemir O, Winkler W: Turkish Heart Study: lipids, lipoproteins, and apolipoproteins. J Lipid Res 1995, 36:839-859. 22. Stene LCM, Giacaman R, Abdul-Rahim H, Husseini A, Norum KR, Holmboe-Ottesen G: Obesity and associated factors in a Pales- tinian West Bank village population. Eur J Clin Nutr 2001, 55:805-811. 23. Fanghanel G, Sanchez-Reyes L, Berber A, Gomez-Santos R: Evalua- tion of the prevalence of obesity in the workers of a general hospital in Mexico. Obes Res 2001, 9:268-273. Acknowledgements 20. Gokcel A, Ozsahin AK, Sezgin N, Karakose H, Ertorer ME, Akbaba M, Baklaci N, Sengul A, Guvener N: High prevalence of diabetes in Adana, a southern province of Turkey. Diabetes Care 2003, 26:3031-3034. g This study was funded, organized and conducted by the Turkish Ministry of Health. The authors express their special thanks to the staff of 14 health screening centers. 21. Martinez-Ros MT, Tormo MJ, Navarro C, Chirlaque MD, Perez- Flores D: Extremely high prevalence of overweight and obes- ity in Murcia, a Mediterranean region in south-east Spain. Int J Obes 2001, 25:1372-1380. Page 5 of 6 (page number not for citation purposes) Page 5 of 6 (page number not for citation purposes) BMC Public Health 2006, 6:92 http://www.biomedcentral.com/1471-2458/6/92 Pre-publication history Satman I, Dinccag N, Yilmaz MT, Sengul AM, Yillar G, Salman S, Sal- man F, Tutuncu Y, Gedik S, Karsidag K, Karadeniz S, Tasyurek A, Sav H: Northern Cyprus: another high prevalence area of diabe- tes and impaired glucose tolerance in the Mediterranean [Abstract]. Diabetelogia 1997, 40:A185. 38 T h l A L P H l hl di i l M di 38. Trichopoulou A, Lagiou P: Healthly traditional Mediterranean diet: an expression of culture, history and lifestyle. Nutr Rev 1997, 55:383-389. Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Page 6 of 6 (page number not for citation purposes) Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Publish with BioMed Central and every scientist can read your work free of charge 39. De Lorgeril M, Salen P, Martin JL: Effect of a meditterranean type of diet on the rate of cardiovascular complications in patients with coronary heart disease. Insights into the cardi- oprotective effect of certain nutriments. J Am Coll Cardiol 1996, 28:1103-1108. 40. Onat A, Surdum-Avci G, Senocak M, Ornek E, Gozukara Y: Plasma lipids and their interrelationship in Turkish adults. J Epidemol Community Health 1992, 46:470-476. y 41. Mahley RW, Mahley LL, Bersot TP, Pépin GM, Palaoglu KE: The Turkish lipid problem: low levels of high density lipoproteins. Turk J Endocr Metab 2002, 1:1-12. 42. Wolf HK, Kuulasmaa K, Tolonen H, Sans S, Molarius A, Eastwood BJ: WHO MONICA Project. Effect of sampling frames on response rates in the WHO MONICA risk factor surveys. Eur J Epidemiol 2005, 20:293-299. 43. Mahley RW, Palaoglu KE, Atak Z, Dawson-Pepin J, Langlois AM, Che- ung V, Onat H, Fulks P, Mahley LL, Vakar F, Ozbayrakci S, Gokdemir O, Winkler W: Turkish Heart Study: lipids, lipoproteins, and apolipoproteins. J Lipid Res 1995, 36:839-859.	15,552
W2042529545.txt_1	Open-Science-Pile	Open Science	Various open science	2,006	Temperature and moisture dependence of soil H<sub>2</sub> uptake measured in the laboratory	None	Unknown	Unknown	4,258	6,989	UC Irvine Faculty Publications Title Temperature and moisture dependence of soil H 2 uptake measured in the laboratory Permalink https://escholarship.org/uc/item/4r7096qk Journal Geophysical Research Letters, 33(14) ISSN 0094-8276 Authors Smith-Downey, Nicole V Randerson, James T Eiler, John M Publication Date 2006-07-01 DOI 10.1029/2006GL026749 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 eScholarship.org Powered by the California Digital Library University of California GEOPHYSICAL RESEARCH LETTERS, VOL. 33, L14813, doi:10.1029/2006GL026749, 2006 Temperature and moisture dependence of soil H2 uptake measured in the laboratory Nicole V. Smith-Downey,1,2 James T. Randerson,3 and John M. Eiler1 Received 28 April 2006; revised 2 June 2006; accepted 13 June 2006; published 25 July 2006. [1] The soil sink of molecular hydrogen is the largest and most uncertain term in the global atmospheric H2 budget. Lack of information about the mechanisms regulating this sink limits our ability to predict how atmospheric H2 may respond to future changes in climate or anthropogenic emissions. Here we present the results from a series of laboratory experiments designed to systematically evaluate and describe the temperature and soil moisture dependence of H2 uptake by soils from boreal forest and desert ecosystems. We observed substantial H2 uptake between 4C and 0C, a broad temperature optimum between 20C and 30C, a soil moisture optimum at approximately 20% saturation, and inhibition of uptake at both low and high soil moisture. A sigmoidal function described the temperature response of H2 uptake by soils between 15C and 40C. Based on our results, we present a framework for a model of the soil H2 sink. Citation: Smith-Downey, N. V., J. T. Randerson, and J. M. Eiler (2006), Temperature and moisture dependence of soil H2 uptake measured in the laboratory, Geophys. Res. Lett., 33, L14813, doi:10.1029/ 2006GL026749. 1. Introduction [2] Molecular hydrogen (H2) is the second most abundant reduced gas in the troposphere (after methane), with a concentration of approximately 530 ppb during the 1990s [Novelli et al., 1999]. Sources of H2 to the troposphere include photolysis of formaldehyde, which is primarily generated from the oxidation of methane and non-methane hydrocarbons, and emissions from fossil fuel combustion and biomass burning. Sinks of hydrogen include oxidation by OH and uptake by soils [Novelli et al., 1999]. The soil sink is the largest and most uncertain term in the H2 budget. Estimating the magnitude of the soil sink has proved challenging and the calculated proportion of H2 uptake by soils to total loss has varied from 62% to 92% [Seiler and Conrad, 1987; Warneck, 1988; Ehhalt, 1999; Novelli et al., 1999; Gerst and Quay, 2001; Hauglustaine and Ehhalt, 2002]. The prospect of a global hydrogen economy has sparked interest in the environmental impact of substantial increases in H2 emissions. A hydrogen economy may be accompanied by reduced NOx emissions, which would increase the atmospheric lifetimes of CH4 and CO due to 1 Geological and Planetary Sciences, California Institute of Technology, Pasadena, California, USA. 2 Also at Engineering and Applied Science, California Institute of Technology, Pasadena, California, USA. 3 Department of Earth System Science, University of California, Irvine, California, USA. Copyright 2006 by the American Geophysical Union. 0094-8276/06/2006GL026749 decreases in OH, and decrease tropospheric O3 levels in polluted urban areas [Schultz et al., 2003]. Tromp et al. [2003] and Warwick et al. [2004] found that replacing fossil fuels with hydrogen fuel cells would lead to a decrease in stratospheric ozone because H2 mixes freely across the tropopause and forms water in the stratosphere. A critical aspect of this problem is the response of the soil sink. Because neither the mechanism nor the environmental controls on soil uptake of H2 are well defined it is difficult to predict both temporal and spatial variability in the magnitude of this sink and how it may respond to future changes in climate or anthropogenic emissions. [3] Schuler and Conrad [1991] studied the temperature response of H2 uptake by soils from a temperate forest, compost bin, and a private garden. They found that at H2 concentrations of 1 ppm, the uptake was optimized at a temperature of 30C whereas at higher H2 concentrations (3000 ppm) H2 uptake was optimized at 65C. At 1 ppm, rates of soil H2 uptake doubled when temperature was increased from 5C to 30C. In an incubation experiment that measured soil uptake for H2 concentrations between 100 and 500 ppm, Trevors [1985] found a lower temperature optimum (20C) for aerobic soils at 60% water holding capacity (WHC) and observed a 3-fold increase in uptake between 5 and 20C. Another experiment by Yonemura et al. [2000b] found a similar temperature optimum, and a factor of 2 difference in uptake rate between 1C and 25C at a single soil moisture level. [4] Previous work suggests that soil moisture also plays an important role in determining the uptake rate of H2. Experiments by Conrad and Seiler [1981; 1985] and Fallon [1982] show that in very dry soils, uptake is stimulated by the addition of water. Godde et al. [2000] measured the H2 oxidation capacity of soils from a temperate forest, agricultural field and meadow in Germany, and found that uptake was generally higher in drier soils (at 30% WHC) than in wetter soils (at 60% WHC). Field studies from a forest and arable field in Japan show that at high soil moisture levels (> 30% volumetric water content) H2 uptake decreases with increasing soil moisture [Yonemura et al., 1999; 2000a], probably from limited H2 diffusion through water filled soils [Yonemura et al., 2000b]. [5] Although several experiments on the temperature and moisture response of H2 uptake by soils exist, there are insufficient data to parameterize a mechanistic model that predicts soil H 2 fluxes across the full range of environmental conditions found in Earth’s major biomes. Such a model would be useful in exploring the sources of variability in the atmospheric H2 record and could be used to predict changes in the H2 cycle due to changing climate or emissions. To construct a mechanistic model of H2 soil uptake, more information is needed about the interaction L14813 1 of 5 L14813 SMITH-DOWNEY ET AL.: TEMPERATURE AND MOISTURE DEPENDENCE L14813 Figure 1. Experimental setup for measuring the temperature and moisture dependence of H2 uptake by soils. Air with known [H2] is pushed through the system from left to right and the outgoing [H2] is measured to determine uptake in the soil column using equation (1). The bubbler hydrated the incoming gas stream slightly, which prevented desiccation of soils over the course of our experiments. The bubbler was removed from the system for the 11% saturated Mojave Desert experiment to prevent addition of water to the soil. The entire beaker apparatus was weighed both before and after each experiment to monitor changes in water content over the course of each experiment. between soil temperature and soil moisture, the sensitivity of soil H2 uptake to soil moisture under unsaturated conditions, rates of soil H2 uptake at temperatures below 0C, and differences in uptake between different biomes. [6] We measured rates of H2 uptake by soils across a wide range of temperatures and soil moisture levels using a flow-through chamber system. In our experiments we used two different surface soils, one from a mature black spruce boreal forest in interior Alaska and another from the Mojave Desert in California. Our experiments imply that there is 1) a strong soil temperature control on H2 uptake, 2) a minimum soil moisture requirement for biological activation, 3) maximized uptake at approximately 20% saturation, 4) decreasing uptake at higher soil moisture levels, 5) substantial H2 uptake occurring between 4C and 0C and 6) that soil type affects the shape of the moisture response. We propose a framework from which global H2 uptake by soils can be modeled based on these data. 2. Methods 2.1. Soil Collection and Preparation [7] Approximately 500 mL of the top 5 cm of soil was collected from a black spruce (Piceae Mariana) forest near Delta Junction, Alaska (63530N, 145440W) and from the University of California Burns Piñon Ecological Reserve in the Mojave Desert (34080N, 116270W). From 1961 to 1990 the mean annual temperature was 2.3C at the boreal forest site and 20C at the Mojave Desert site. Mean annual precipitation was 30 cm/yr at the boreal forest site and 11 cm/yr at the Mojave Desert site (Western Regional Climate Center (WRCC) station observations from Big Delta, AK and Twentynine Palms, CA, available online at http://www.wrcc.dri.edu/clim-sum.html). Soils were stored in sealed containers for transport back to the lab, after which they were stored at room temperature in jars sealed with Parafilm. The carbon and nitrogen content of the boreal forest soil was 38.5% C and 1.11% N whereas the Mojave Desert soil was 0.3% C and 0.02% N (measured with an elemental analyzer, Carlo Erba, Lakewood, NJ). 2.2. Flux Measurements [8] Hydrogen calibration gases between 5 and 2000 ppb H2 were generated by diluting a standard gas containing 5070 ± 25 ppm H2 (Scott-Marrin, Riverside, CA) with ultrapure N2. Samples from cylinders of compressed air were measured following calibration of the instrument and these cylinders were used as secondary standards throughout the experiment. [9] We created a flow through apparatus for measuring H2 uptake by insetting a 1=2 00 OD glass tube through a 2L beaker and filling the tube with soil (Figure 1). Gas from a compressed air cylinder containing 1585 ± 21 ppb H2 flowed, in order, through a mass flow controller (MKS, Wilmington, MA), through a flask filled with water to control humidity, through the 1=2 00 glass tube filled with the soil sample, through a dry ice loop to remove moisture and finally to the inlet of a TA 3000R Reducing Gas Analyzer (Ametek Process Instruments, Newark, DE) at a rate of 10 standard cubic centimeters per minute (sccm). The TA3000R RGA is a continuous flow instrument with a Unibead 1S and an MS 13X column for separation of H2 and CO. Ultra-torr fittings (Swagelok, Solon, OH) were used at each joint, and the system was leak tested with an 80% H2 source. The TA 3000R instrument measures H2 with a relative precision of 1.5%. 2.3. Temperature and Moisture Experiments [10] In preparation for the temperature and moisture manipulation experiments, soils were thinly spread out in plastic pans and allowed to dry at room temperature (23C) overnight. Soil samples were then split into 5 equal portions and water was added to each sample in increments. In these experiments, soil moisture was described as ‘% saturated’, which was calculated as the ratio of water (g) contained in soils to the amount contained after soils are fully submerged in pure H2O and allowed to drain for 10 minutes. The soils were stirred to evenly incorporate the added water and stored in 50 mL test tubes sealed with Parafilm for at least 3 days before experiments began. The manipulation of the soils may have changed the soil porosity and existing 2 of 5 L14813 SMITH-DOWNEY ET AL.: TEMPERATURE AND MOISTURE DEPENDENCE L14813 and T is the temperature (K). Pressure was assumed to be 1.01325 105 Pa. 3. Results Figure 2. Time series of [H2]out from a single laboratory experiment using the boreal forest soils at 23% saturation. The difference between the 23C no soil ([H2]in) and [H2]out was used in equation 1 to calculate the uptake of H2 in the soil column. The drop in [H2]out between 4C and 15C corresponds with a warm temperature excursion due to the exchange of the salt-water solution from the beaker with an ethanol/ice solution. microbial communities and, for this reason, our results may not be representative of in situ fluxes. [11] For each soil moisture level, the column was filled with 10 mL of soil with a cone of filter paper at each end. The beaker was filled with water of different temperatures, and the concentration of outgoing H2 was measured every 3.5 minutes. Temperatures were cycled through in the following order 23, 30, 37, 10, 0, 4, 15C (Figure 2). We used a hot plate to maintain 30C and 37C, a water bath with periodic ice additions to maintain 10C, an ice water bath to maintain 0C, an ice/salt water bath to maintain 4C, and an ice/ethanol/water bath to maintain 15C. The water in the beaker was drained and replaced between each temperature level to ensure a quick transition. We used a stir bar to avoid temperature stratification within the beaker. We conducted one set of temperature manipulations on soils with a thermocouple embedded in the soil and another in the water bath. The temperature of the soil equilibrated with the water bath temperature within minutes, and there was no observed temperature offset between the soil and the water bath. The thermocouple wire could not be placed in the soil during the flow through experiments because a gas-tight seal was necessary at each joint. Each temperature/moisture level was considered stable after the outgoing H2 concentration had remained constant for at least 30 minutes (e.g., Figure 2). 2.4. Uptake Rate Estimates 1 [12] The uptake rate (pmol cm3 soil s ) of each soil sample was calculated using the measured difference between the incoming and outgoing H2 concentrations: H2 uptake ¼ ½H2 in ½H2 out f P Vsoil R T 1 109 ð1Þ [ 13 ] Our experiments showed a dependence of H 2 uptake on soil moisture and temperature in both soils (Figure 3). The maximum flux from the boreal forest soil (0.8 pmol cm3 soil s1) was approximately twice as large as the maximum flux rate from the Mojave Desert soil (0.35 pmol cm3 soil s1). We observed a broad temperature maximum between 10C and 35C (Figure 3). Substantial uptake continued at 4C (especially for intermediate soil moisture levels), but was nearly eliminated by 15C. The uptake rate increased sharply with increasing temperature between 4C and 0C for the boreal forest soils at intermediate saturation levels, whereas the changes in uptake rate with temperature were more gradual in the Mojave Desert soils. In a similar experiment on soils from a California forest, fluxes decreased substantially when the temperature was increased to 65C (Figure S11) suggesting uptake inhibition at high temperatures. [14] We observed both high and low soil moisture inhibition of H 2 uptake with soils from both sites (Figure 3). For the boreal forest soils, increasing the saturation level from 4% to 10% caused uptake to increase by 300% at 23C. Similarly, increasing the saturation from 11% to 19% for the Mojave Desert soils caused the uptake rate to increase from approximately zero to the maximum uptake levels (Figure 3). The boreal forest soils continued consuming substantial H2 at higher saturation levels than the Mojave Desert soils, and did not show evidence of moisture inhibition until 59% saturation. [15] We constructed normalized contour plots of the boreal forest and Mojave Desert fluxes as a function of both temperature and soil moisture (Figure 4). Contour values ranged between 0 and 1 and were scaled by the maximum observed flux at each site. The maximum flux occurred at 23C and 23% saturation for the boreal forest soil, and at 30C and 19% saturation for the Mojave Desert soil. [16] The response to temperature, when normalized to the maximum flux at each soil moisture level, was similar between the two sites and can be described by a sigmoidal function with the following parameters (Figure S2): f ðT Þ ¼ 1 1 þ expð0:1718 T þ 46:938Þ ð2Þ where T is the temperature (K). This function is valid for temperatures between 15C and 40C (with an r2 of 0.72). At temperatures above 40C, our results suggest that uptake decreases with increasing temperature (Figure S1). [17] The moisture response cannot be characterized by a simple function, largely due to the effect of soil moisture on the diffusion of H2 into soils, which varies considerably with soil type. It is clear, however, that a minimum level of soil moisture must be attained before uptake is possible. To account for the effects of soil moisture as it fills pore space, where f is the flow rate (cm3/s), P is the pressure (Pa), Vsoil is the volume of soil (cm3), R is the gas constant 3 of 5 1 Auxiliary material is available in the HTML. doi:10.1029/2006GL026749. L14813 SMITH-DOWNEY ET AL.: TEMPERATURE AND MOISTURE DEPENDENCE L14813 R is the gas constant, and T is temperature (K). We assumed that the reaction was first order based on chamber measurements of H2 uptake in the field. This framework can be used to estimate H2 uptake as a function of soil moisture and temperature. 4. Discussion and Conclusions [18] We systematically defined the temperature and moisture controls on soil H2 uptake over a wide range of environmental conditions and identified, for the first time, uptake at subzero temperatures (from 4C to 0C). Our results provide additional evidence that a minimum moisture level is required for biological activation of microbes utilizing H2 [e.g., Conrad and Seiler, 1981; Fallon, 1982; Conrad and Seiler, 1985], but that once pore spaces are filled with water, diffusion of H2 into the soil becomes limited and fluxes decrease [e.g., Yonemura et al., 2000b]. Figure 3. Hydrogen uptake rates as a function of temperature for different soil moisture levels. (a) Boreal forest soils collected near Delta Junction, AK and (b) Mojave Desert soils collected near Yucca Valley, CA. Note that the scale in Figure 3a is twice that of Figure 3b. Uptake rate at 23C was measured twice for the boreal forest soils at 10% and 23% saturation, and in both cases the difference between measurements was less than 5%. pﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ Standard deviations were calculated as s2in þ s2out where sin and sout are the standard deviation of the measurement of incoming and outgoing [H2] respectively. The negative values observed probably do not represent an actual production of H2, but illustrate the level of internal noise in our measurement system. hydrogen uptake must be modeled as a diffusion process where the uptake rate is scaled according to the results presented here: ls ¼ f ðT Þgð M Þ * lmax * ½H2 * P R*T ð3Þ where f(T) and g(M) are functions of soil temperature (T) (Equation 2) and moisture (M) respectively that range between 0 and 1, lmax is the maximum possible H2 uptake (1/s) under ideal soil temperature and moisture conditions (i.e., when f(T) and g(M) are equal to 1), [H2] is the available hydrogen concentration (ppb), P is pressure (Pa), Figure 4. Contour plots of normalized uptake rate at each site as a function of temperature and moisture. Data were normalized by the maximum flux at each site. Grey dots represent locations of the data points used to generate the contour plots. (a) For boreal forest soils there was a relatively strong gradient between 0% and 10% saturation and 4C and 0C. (b) For Mojave Desert soils there was more symmetry about the moisture maximum as compared to the boreal forest soils. In constructing this plot, we assumed negative flux values measured in the Mojave Desert soils (Figure 3b) were equal to zero. 4 of 5 L14813 SMITH-DOWNEY ET AL.: TEMPERATURE AND MOISTURE DEPENDENCE The low soil moisture inhibition of H2 uptake has important implications for desert soils, and may lead to maximum H2 uptake in wetter months despite the possibility of lower temperatures. [19] The shape of the temperature and moisture response (Figure 4a) suggests that in boreal forest soils most of the variability in H2 uptake by soils occurs under relatively low (4C to 15C) temperatures and low soil moisture conditions. These observations may have important implications for the seasonal cycle of H2 uptake globally. Northern soils routinely experience shifts in temperature over the range 4C to 15C seasonally. The timing of the spring drawdown of H2, (beginning in May) and buildup (beginning in October) at northern latitudes [Novelli et al., 1999] may be sensitive to the sharp increase in uptake that we observed between 4C to 15C (Figure S2). It is possible for northern soils to consume H2 through the warmer parts of the winter, although the effect of snow cover, which may limit diffusion of H2 into soils, must be measured and accounted for. Additionally, the seasonal and interannual soil moisture changes at these latitudes will affect the timing and magnitude of H2 uptake. [20] Boreal forest soil fluxes are approximately twice as large as Mojave Desert soil fluxes at the same temperature and soil moisture content. We hypothesize that this is due, in part, to differences in organic carbon content and soil structure. The two soils we examined have carbon contents near the extremes observed in common soils (0.02 to 38.5%) but more data is needed to clearly define the dependence of uptake on organic carbon content of soils. Although our laboratory measurements provided evidence that on a volumetric basis, boreal soils consume more H2 than desert soils, chamber observations at these two locations do not show a large difference between surface flux rates per m2 ground area [Rahn et al., 2002] (N. SmithDowney et al., manuscript in preparation, 2006). The net uptake at the surface is a product of the maximum uptake capacity of a soil (lmax), the temperature and moisture controls defined here, along with the diffusion of H2 into the soil profile. It is possible that despite a higher capacity for H2 uptake per cm3 soil, the boreal forest uptake is diffusion limited due to higher soil moisture or differences in soil structure. Constructing a model that takes all of these factors into account will allow us to examine sources of variability in the observed atmospheric H2 record and to assess how climate change will influence future atmospheric levels of H2. [21] Acknowledgments. NSD thanks J. Leadbetter for lab and instrument use, K. Treseder for collecting soils in Alaska, X. Xu for assistance with the elemental analyzer, P. Ghosh, J. O’Leary, H. Afek and M. Child for help in the lab, and N. Downey. NSD received support from the NCER L14813 STAR program, EPA. JTR acknowledges support from UC Irvine’s School of Physical Sciences Dean’s Innovation Fund. This work was also supported by a generous gift from General Motors and William Davidow and family. References Conrad, R., and W. Seiler (1981), Decomposition of atmospheric hydrogen by soil microorganisms and soil enzymes, Soil Biol. Biochem., 13, 43 – 49. Conrad, R., and W. Seiler (1985), Influence of temperature, moisture, and organic carbon on the flux of H2 and CO between soil and atmosphere: Field studies in tropical regions, J. Geophys. Res., 90, 5699 – 5709. Ehhalt, D. H. (1999), Gas phase chemistry of the troposphere, in Global Aspects of Atmospheric Chemistry, edited by R. Sellner, pp. 21 – 109, Springer, New York. Fallon, R. D. (1982), Influences of pH, temperature, and moisture on gaseous tritium uptake in surface soils, Appl. Environ. Microbiol., 44, 171 – 178. Gerst, S., and P. Quay (2001), Deuterium component of the global molecular hydrogen cycle, J. Geophys. Res., 106, 5021 – 5031. Godde, M., et al. (2000), Hydrogen consumption and carbon monoxide production in soils with different properties, Biol. Fertil. Soils, 32, 129 – 134. Hauglustaine, D. A., and D. H. Ehhalt (2002), A three-dimensional model of molecular hydrogen in the troposphere, J. Geophys. Res., 107(D17), 4330, doi:10.1029/2001JD001156. Novelli, P. C., et al. (1999), Molecular hydrogen in the troposphere: Global distribution and budget, J. Geophys. Res., 104, 30,427 – 30,444. Rahn, T., J. M. Eiler, N. Kitchen, J. E. Fessenden, and J. T. Randerson (2002), Concentration and dD of molecular hydrogen in boreal forests: Ecosystem-scale systematics of atmospheric H2, Geophys. Res. Lett., 29(18), 1888, doi:10.1029/2002GL015118. Schuler, S., and R. Conrad (1991), Hydrogen oxidation activities in soil as influenced by pH, temperature, moisture and season, Biol. Fertil. Soils, 12, 127 – 130. Schultz, M. G., et al. (2003), Air pollution and climate-forcing impacts of a global hydrogen economy, Science, 302, 624 – 627. Seiler, W., and R. Conrad (1987), Contribution of tropical ecosystems to the global budgets of trace gases, especially CH4, H2, CO and N2O, in The Geophysiology of Amazonia: Vegetation and Climate Interactions, edited by R. E. Dickerson, pp. 33 – 62, John Wiley, Hoboken, N. J. Trevors, J. T. (1985), Hydrogen consumption in soil, Plant Soil, 87, 417 – 422. Tromp, T. K., et al. (2003), Potential environmental impact of a hydrogen economy on the stratosphere, Science, 300, 1740 – 1742. Warneck, P. (1988), Chemistry of the natural atmosphere, in International Geophysics Series, vol. 41, p. 757, Elsevier, New York. Warwick, N. J., S. Bekki, E. G. Nisbet, and J. A. Pyle (2004), Impact of a hydrogen economy on the stratosphere and troposphere studied in a 2-D model, Geophys. Res. Lett., 31, L05107, doi:10.1029/2003GL019224. Yonemura, S., et al. (1999), Continuous measurements of CO and H2 deposition velocities onto an andisol: Uptake control by soil moisture, Tellus, Ser. B, 51, 688 – 700. Yonemura, S., S. Kawashima, and H. Tsuruta (2000a), Carbon monoxide, hydrogen, and methane uptake by soils in a temperate arable field and a forest, J. Geophys. Res., 105, 14,347 – 14,362. Yonemura, S., et al. (2000b), Model analysis of the influence of gas diffusivity in soil on CO and H2 uptake, Tellus, Ser. B, 52, 919 – 933. J. M. Eiler and N. V. Smith-Downey, Geological and Planetary Sciences, California Institute of Technology, MS 100-23, Pasadena, CA 91125, USA. (nicolev@gps.caltech.edu) J. T. Randerson, Department of Earth System Science, University of California, Irvine, Croul Hall, Irvine, CA 92697 – 3100, USA. (jranders@uci.edu) 5 of 5.	3,626
https://openalex.org/W4360923793	OpenAlex	Open Science	CC-By	2,022	The Effect of Salty Groundwater Levels on the Grass Yield of Some Pasture Crops and Soil Salinity	Barış Bahçeci	English	Spoken	6,687	11,569	ABSTRACT the second year. According to the Duncan test results, it was determined that the H1 (40 cm groundwater depth) treatment in the first year provided the highest yield in all three cultivars and formed the first group (p<0.01) in the Duncan test. The interaction between water table depths and plant species was statistically significant at the p<0.05 level. At the end of the experiment, the salt concentration of the topsoil (40 cm depth) increased significantly (p=0.025) at all water table depths. Moreover, although the sodium adsorption ratio of the inlet water was low, it was determined that the exchangeable sodium percentage of the soils increased significantly at all groundwater levels at the end of the trial. Salinity is one of the most serious environmental factors limiting the yield of plants. Because many crops experience yield losses due to the harmful effects of high salt content in soil and water. The increase in the area of land affected by salt has the potential to create problems in terms of food safety. In this context, it is necessary to develop some cultural practices to prevent or reduce the harmful effects of salinity. This study investigated the effects of salty groundwater at different depths on the yield and soil salinity of some forage crops grown in semi-arid regions for three years. The experiment was conducted using the randomized block split-plot design. The effect of water table depths on the yields of the cultivars in the first year was found to have a statistically significant (p<0.01) effect while it was insignificant in Keywords: Forage crops, Pasture lands, Festuca elatior, Lotus strictus, Puccinella distans, Groundwater salinity The Effect of Different Depths of Salty Groundwater on Yield and Soil Salinity of Some Pasture Crops aDepartment of Agricultural Structure and Irrıgation, Faculty of Agriculture, Harran University, Şanlıurfa, Türkiye bDepartment of Agricultural Structure and Irrıgation, Faculty of Agriculture, Harran University, Şanlıurfa, Türkiye (Retired) Barış BAHÇECİa , Ali Fuat TARIa , İdris BAHÇECİb* aDepartment of Agricultural Structure and Irrıgation, Faculty of Agriculture, Harran University, Şanlıurfa, Türkiye bDepartment of Agricultural Structure and Irrıgation, Faculty of Agriculture, Harran University, Şanlıurfa, Türkiye (Retire ARTICLE INFO Research Article Corresponding Author: İdris BAHÇECİ, E-mail: bahceci@harran.edu.tr Received: 2 May 2021 / Revised: 18 Oct 2022 / Accepted: 10 Oct 2022 / Online: 25 Mar 2023 JAS JAS JAS Cite this article i BAHÇECİ B, TARI A F, BAHÇECİ İ (2023). The Effect of Different Depths of Salty Groundwater on Yield and Soil Salinity of Some Pasture Crops. Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 29(2):546-554. DOI: 10.15832/ankutbd.918734 Journal of Agricultural Sciences (Tarim Bilimleri Dergisi) Journal of Agricultural Sciences (Tarim Bilimleri Dergisi) 2023, 29 (2): 546-554 DOI: 10.15832/ankutbd.918734 Journal of Agricultural Sciences (Tarim Bilimleri Dergisi) J Agr Sci-Tarim Bili e-ISSN: 2148-9297 2023, 29 (2): 546-554 1. Introduction For example; Puccinella distans can provide a good yield of up to 26 dS m-1 salinity in salty soils (Öztan 1965; Akhazari et al. 2012). Bennett & Barrett-Lennard (2013) reported that samphire (Tecticornia pergranulata) and Puccinellia (Puccinellia ciliata) grow in areas with a salinity of 16 dS m-1. The increased salinity causes only a decrease in plant height (Ashkan & Jalal 2013; Kuşvuran et al. 2014). A. elongatum species are less damaged by salinity conditions than other Agropyron species because it is more adaptable to salinity conditions in morphological terms (Koç & Acar 2017). In addition, Agropyron species (Agropyron spp.) can naturally grow in areas with salinity issues in the Central Anatolia rangeland (Acar et al. 2016). The grass yield of 70% of the natural pastures in the Konya plain is very low, and the average dry grass yield is approximately 200 kg ha-1. High-quality forage plants with high grass yields have been replaced by sour marsh plants such as Carex and Juncus (Tosun 1967). Tall wheatgrass (A. elongatum) and crested wheatgrass (A. cristatum) from Agropyron spp. give a yield with a 50% loss in high salinity levels, which is not available to grow other plant species. (Akhazari et al. 2012; Ashkan & Jalal 2013). The presence of saline groundwater at 25 cm depth had a detrimental effect on the production of biomass and its components on Rhodes grass (Chloris gayana), whereas the effect at 125 cm and greater depths was neutral (Chiacchiera et al. 2016). Leptochloa fusca, Spartina patens, and Sporobolus virginicus (Smyrna) have been reported to be promising halophytic plants for feeding goats and sheep in desert areas (Ashour et al. 1997). For the reasons mentioned above, this study was carried out to determine the effects of salty groundwater at different depths on the yield and soil salinization of some forage crops in pasture areas in semi-arid regions and to contribute to the determination of forage plant varieties suitable for semi-arid conditions. 1. Introduction Flatlands in arid and semi-arid regions and alluvial plains on the coasts have inadequate drainage due to topographic and soil structure. These lands have been salted by high groundwater levels and insufficient drainage. This type of land that covers extremely large areas of the world, is in the form of pastures or abandoned lands with low, and poor-quality grass yields. These unproductive pastures were degraded by negative factors such as overgrazing, erosion, drainage, and barrenness. These lands have reduced plant diversity due to the high-water table and excessive salt, and natural vegetation has become poor-quality pastures dominated by barren-halophyte plants of low nutritional value that are not liked by animals. In many countries where livestock is based on meadow and pasture farming, it is aimed at producing cheap and healthy food by growing salt-resistant forage plants and their mixtures in meadows and pastures under the influence of high groundwater (Dieleman 1977). Establishing artificial pastures consisting of acceptable forage crops, currently used as pastures, may significantly increase the agricultural potential of these lands. Another way to benefit from these lands is to rehabilitate them. However, although salt-affected soils are chemically recovered, it takes a long time to restore the physical properties of these soils. Therefore, by cultivating some salt-tolerant forage crops during this period, both the recovery of the physical properties of the soil and the farm income will be contributed. Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 In this context, halophytes are important because of the prevalence around the Lake Tuz and arid soils in central Anatolia (Birand, 1961). For example; Puccinella distans can provide a good yield of up to 26 dS m-1 salinity in salty soils (Öztan 1965; Akhazari et al. 2012). Bennett & Barrett-Lennard (2013) reported that samphire (Tecticornia pergranulata) and Puccinellia (Puccinellia ciliata) grow in areas with a salinity of 16 dS m-1. The increased salinity causes only a decrease in plant height (Ashkan & Jalal 2013; Kuşvuran et al. 2014). A. elongatum species are less damaged by salinity conditions than other Agropyron species because it is more adaptable to salinity conditions in morphological terms (Koç & Acar 2017). In this context, halophytes are important because of the prevalence around the Lake Tuz and arid soils in central Anatolia (Birand, 1961). 2.3. Method Trials were carried out with 3 forage plants, A- Lotus sitrictus, B- Puccinella distans, C- Festuca elatior and 4 groundwater depth levels, H1=40 cm H2=60 cm H3=80 cm H4=110 cm. 2.3.1. Setting up the experiment The experiment was carried out in 3 replicated, randomized blocks split plots trial design. The experiment was conducted in cylindrical steel barrels with a depth of 120 cm and with a diameter of 56 cm. The plot areas are 0.2462 m2 for planting and harvesting. In the first place, the volume weights of the soils at the trial site were determined by the layers of 20 cm. The soils were dug, stored, and allowed to dry in the shade, depending on the soil layers. Ten cm thick sand gravel was laid on the bottoms of the barrels. To measure the depth, 10 cm of soil was placed on the top of the sand-gravel layer. It was compacted until the original bulk weight was achieved with a mallet. Then the second 10 cm layer was laid, and the same operations continued until the barrel was full. To create a water table in the barrels, holes in the specified depth are drilled and the outlet drainage pipes were located. Groundwater was pumped through inlet pipes placed under the tanks to a 4 m high water tank. The salt content of the water entered and discharged from the barrels was measured every month. 2.3.2. Properties of the plants used in the experiment Festuca elatior, Puccinellia distans, and Lotus strictus, which are under the shallow water table around Konya in the natural vegetation of Aslım pasture lands, were used in the experiment (Figure 2). Festuca elatior; high meadow ball, salt resistant, 40-60 cm tall, also called meadow ball (Tosun 1974). Its multi-sibling, bundles are 30 cm in diameter, growing vertically upwards, between 20 cm and 1 meter in length. Leaves are curled, cylindrical leaf sheathed. The underside of the leaf is brightly colored. Flowers are unique with many large and elongated spikelets together to form compound clusters. Puccinellia distans is a green-colored forage crop with wheatgrass spikes resistant to salt (Werner & Senghas 1973). Five-forty cm in length with body characteristics of herbaceous plants native to Turkey. It is common in semi-marshy areas around inland salty lakes. 2.2. Climate The Konya Plain has the typical characteristics of the continental climate; the summers are dry and hot, and the winters are snowy and cold. The highest annual temperature average is 11.6 ºC, the annual precipitation is 327.7 mm and the average relative humidity is 63% (Anonymous, 2018). During the trial, precipitation was 386 mm in the first year, and 277.8 mm in the second year, the evaporation was 1,171 in the first year and 1,243 mm in the second year and the average temperature was 11.1 in the first year and 10.5 ºC in the second year. 2.1. Trial site The trial site is the salty lands in Konya Plain with a high-water table. These lands are widely located in different parts of the Central Anatolia (Figure 1), especially around Lake Salt, in Aslım pastures, in the plains of Karapınar, Ereğli plain, and around Lake Hotamış (Meester 1970). Figure 1- The geographical location of the research site The trial site soils are in the hydromorphic soil group, with medium depth, it is flat land (Atalay & Secerli 1971). The upper soil is silty loam, and the lower soil is silty and organically rich. Its lime content is high, it is excessively salty and salt content decreases with depth (Table 1). The water conductivity of light sodium soils is in the middle group of the upper layers and the medium-slow group of the lower layers (Meester 1970). Figure 1- The geographical location of the research site The trial site soils are in the hydromorphic soil group, with medium depth, it is flat land (Atalay & Secerli 1971). The upper soil is silty loam, and the lower soil is silty and organically rich. Its lime content is high, it is excessively salty and salt content decreases with depth (Table 1). The water conductivity of light sodium soils is in the middle group of the upper layers and the medium-slow group of the lower layers (Meester 1970). 547 Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 Table 1- Some physical and chemical properties of trial soils Soil depth cm Texture pH ECe NaX CEC ESP Lime OM S Si C dS m-1 cmol+ kg-1 % % % 0-20 24.5 68.7 6.7 Si 7.8 17 3.1 13 24 22.0 4.35 20-40 20.7 70.3 9.0 Si 7.0 15 2.3 10.9 21 23.3 2.90 40-70 8.0 87.9 4.0 Si 6.5 12 1.4 11.4 13 41.7 2.56 70-110 13.3 87.0 0.0 Si 6.4 6.9 0.3 8.5 3.5 43.1 2.04 ECe: Electrical conductivity of soil extract, NaX: Exchangeable Na, CEC: Cation exchange capacity, ESP: Exchange Na percentage, OM: Organic material, S: Sandy, Si: Silty, C: Clay 2.2. Climate Table 1- Some physical and chemical properties of trial soils 2.3. Method It generally grows in sandy loam, silty loam, and loamy soils and has a strong root system and anti-erosion feature, which is important for grazing in salty areas. Lotus strictus is a salt-resistant, yellow-flowered forage plant that reproduces with the rhizome and is 50-100 cm tall and highly branched (Kyell Vist 1971). Glabrous, with short hair only on top, perennial, erect, thick, woody below, with umbrella-like inflorescences bloom in August - September. 548 Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 2.3.5. Statistical analysis Grass yields of the cultivars were analyzed in randomized blocks with a split-plot design. The effect of groundwater on salt accumulation was evaluated through regression analysis and pairwise comparisons (Yurtsever 1984). 2.3.4. Sowing, fertilizing, and harvesting The seeds were sown by hand, but as there was not adequate germination, the Lotus strictus plots were replanted. Before planting, 40 kg N, and 100 kg P2O5 were given per hectare and mixed into the soil. During the summer, the trial plots were irrigated 3 times with groundwater. Figure 2- Pasture plants used in the experiment Figure 2- Pasture plants used in the experiment When the plants reached grazing maturity, Festuca elatior, and Agropyron elongatum 20 cm, Puccinella distans, and Lotus strictus 15 cm in length, were harvested, and wet and dry grass weights were determined. When the plants reached grazing maturity, Festuca elatior, and Agropyron elongatum 20 cm, Puccinella distans, and Lotus strictus 15 cm in length, were harvested, and wet and dry grass weights were determined. In order to determine the dry grass yield, the wet grass samples were dried in an oven at 65 ºC until they reached a constant weight, and their moisture content was determined. In order to determine the dry grass yield, the wet grass samples were dried in an oven at 65 ºC until they reached a constant weight, and their moisture content was determined. 3.1. Grass yields of crop varieties The highest yield was obtained for 40 cm water table depth. The dry grass yields of F. elatior, P. distans, and L. strictus are 744, 642, and 615 kg da-1, respectively. When the groundwater level fell to 60 cm depth, the hay yields decreased to 407, 383, and 454 ka da-1, respectively. The effect of water table levels on plant yield was found to be significant, and H1=40 cm water table depth formed the first group (F0.01=34.57 and p<0.01) in both years, and also the interaction between water table and plant species (p=0.05) was found to be significant (Table 2) and crop varieties (A, B, C). Table 2. Dry grass yields versus different water table depths (kg da-1) Groundwater depth, cm Crop varieties: 1st year Average Crop varieties: 2st year Average A B C A B C 40 744 642 615 667a 816 173 494 494a 60 407 383 454 415ab 543 72 378 331b 80 214 318 334 289b 792 125 209 375b 110 224 270 314 269b 509 99 206 271c Table 2. Dry grass yields versus different water table depths (kg da-1) However, while L. strictus regenerated itself with rhizome and maintained its productivity at all water levels, the decrease in the yield of P. distans and F. elatior was significant statistically (Figure 3). Due to a decrease from 60 cm to 110 cm in the water table, the efficiency decrease continued, but the decrease was not statistically significant. In these pasture areas, a groundwater depth of 40 cm should be considered a critical level. It should be noted that increasing the depth of the water table, in addition to damaging the existing vegetation due to water stress, may also accelerate the oxidation of organic matter in the upper soil layers. In these pasture areas, a groundwater depth of 40 cm should be considered a critical level. It should be noted that increasing the depth of the water table, in addition to damaging the existing vegetation due to water stress, may also accelerate the oxidation of organic matter in the upper soil layers. 549 Bahçeci et al. 3.1. Grass yields of crop varieties Tall wheatgrass has less tolerance to salinity, and the yield is severely reduced (by 55%) under high salinity (Grattan et al. 2004, Robinson et al. 2004). To maintain high bermudagrass yields, it is recommended that soil salinities should not exceed Ece 12 dS m-1 (Kaffka et al. 2004). Many halophytic plants thrive optimally at 20 to 25 g l-1 soil salinity, and at 30 to 40 g l-1 show 25% to 50% growth reduction but they can sustain their lives in soils containing 60 to 90 g l-1 salt (Miyamoto et al. 1994). Leptochloa fusca, Spartina patensand, Sporobolus virginicus (Smyrna) are shown as promising halophytic plants for feeding goats and sheep in desert lands using available saline water for irrigation areas (Ashour et al. 1997). In a land with a drainage problem, 85% of the yield from tall wheatgrass was obtained from forage plants irrigated with saline drainage water, while the yield for alfalfa was 43%. (Suyama et al. 2007). Tall wheatgrass has less tolerance to salinity, and the yield is severely reduced (by 55%) under high salinity (Grattan et al. 2004, Robinson et al. 2004). To maintain high bermudagrass yields, it is recommended that soil salinities should not exceed Ece 12 dS m-1 (Kaffka et al. 2004). Many salt-tolerant forage crop species, such as the forage crops discussed in this study, can be grown successfully in semi-arid conditions dominated by shallow groundwater. On lands with similar conditions, artificial pastures can be established before improvement. Puccinellia distans is a perennial cool-season salty grass, that can adapt to arid climates and extremely saline-alkaline soil conditions (Hughes 1972; Brotherson 1987; Scalia et al. 2009; Ehsani et al. 2016), and, spread in salty meadow-pasture areas. This plant produces between 4 and 10 tons of hay per hectare per year (Warren et al. 1994) and this delicious species (Shidaei & Namati 1978) is generally consumed by sheep [Peng et al. 2004; Akhazari et al. 2012, Ashkan & Jalal 2013], but, it has been reported that increasing salinity causes a decrease in plant height (Acar et al. 2016). Considering that 0.4 hectares of artificial pasture per sheep provides the most economical live weight gain in the shallow water table conditions of A. elongatum, P. capillaris and F. elatior in this research area (Uçar 1982), the usefulness of such an application can be easily understood that it can make a significant contribution to increasing feed production. 3.1. Grass yields of crop varieties - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 Figure 3 Dry grass yields versus water table depths in different years Figure 3- Dry grass yields versus water table depths in different years h correlation coefficients of the relationship between the decrease in the water table level and the grass yields indicated that n reason for the decrease in the crop was water stress. Although similar results were obtained in the second year of the trial, ative effects of the increase in soil salinity were also observed However although L strictus regenerated by rhizome and Figure 3- Dry grass yields versus water table depths in different years The high correlation coefficients of the relationship between the decrease in the water table level and the grass yields indicated that the main reason for the decrease in the crop was water stress. Although similar results were obtained in the second year of the trial, the negative effects of the increase in soil salinity were also observed. However, although L. strictus regenerated by rhizome and maintained its productivity at all water levels, there were significant decreases in the yield of P. distans and F. elatior (Figure 3). The high correlation coefficients of the relationship between the decrease in the water table level and the grass yields indicated that the main reason for the decrease in the crop was water stress. Although similar results were obtained in the second year of the trial, the negative effects of the increase in soil salinity were also observed. However, although L. strictus regenerated by rhizome and maintained its productivity at all water levels, there were significant decreases in the yield of P. distans and F. elatior (Figure 3). Many halophytic plants thrive optimally at 20 to 25 g l-1 soil salinity, and at 30 to 40 g l-1 show 25% to 50% growth reduction but they can sustain their lives in soils containing 60 to 90 g l-1 salt (Miyamoto et al. 1994). Leptochloa fusca, Spartina patensand, Sporobolus virginicus (Smyrna) are shown as promising halophytic plants for feeding goats and sheep in desert lands using available saline water for irrigation areas (Ashour et al. 1997). In a land with a drainage problem, 85% of the yield from tall wheatgrass was obtained from forage plants irrigated with saline drainage water, while the yield for alfalfa was 43%. (Suyama et al. 2007). 3.2. Change in soil salinity and sodium The salt distribution in the soil profile was similar for all groundwater levels. The distribution of salts in the soil profile was typical, as in soils under the influence of a high water table. That is, the salt concentrations were decreased from the upper layers to the lower 550 Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 layers. Statistical analysis after the experiment showed that the interaction of soil layers and water levels was significant (F0.01=34.57 p<0.01) in the first year. Analysis by year showed that the effect of water levels on salinization was insignificant (F0.01=124.44 p<0.01), but salt accumulation in soil layers was significantly different. layers. Statistical analysis after the experiment showed that the interaction of soil layers and water levels was significant (F0.01=34.57 p<0.01) in the first year. Analysis by year showed that the effect of water levels on salinization was insignificant (F0.01=124.44 p<0.01), but salt accumulation in soil layers was significantly different. In other words, the water levels were effective in the salinization of the soil layers. In the beginning, the topsoil layer had the highest salt content and the salinity have been decreased with increasing depth (Table 3, Figure 4). Differences in soil salt contents between treatments were insignificant (p=0.202>0.05). However, there was a statistically (p<0.05) significant difference between the salt contents of the soil layers. 3.2. Change in soil salinity and sodium Table 3- Variation of salinity in soil layers versus different water table depths and crop varieties (A, B, C) Groundwater depth, cm Soil Before trial After trial depth, ECe, dSm-1 ECe, dSm-1 A B C Mean A B C Mean 40 0-40 15.3 14.8 15.4 15.2 18.3 21.2 24.7 21.4 40-70 10.8 6.9 8.5 8.6 7.9 9.9 12.4 10.1 70-110 5.9 9.0 9.4 8.1 4.9 6.7 6.4 6.0 60 0-40 14.7 16.7 14.5 15.3 20.3 28.8 23.4 24.2 40-70 8.8 14.3 12.9 12.0 13.1 14.0 16.6 14.6 70-110 6.7 10.0 6.0 7.6 8.1 7.8 9.2 8.4 80 0-40 16.7 16.4 16.4 16.5 13.9 17.3 12.4 14.5 40-70 11.1 10.9 12.0 11.3 13.3 9.7 11.0 11.3 70-110 6.3 7.0 6.3 6.5 6.4 6.9 8.9 7.4 110 0-40 16.5 15.8 16.6 16.3 19.3 16.7 17.7 17.7 40-70 10.8 7.7 9.2 9.0 14.3 9.6 11.5 11.8 70-110 7.1 6.7 6.7 6.8 10.4 6.7 8.1 8.4 Before and after the trial all soil layers and all treatments, p=0.0036 <0.01 Before and after the trial topsoil layer and all treatments, p=0.025 <0.05 Table 3- Variation of salinity in soil layers versus different water table depths and crop varieties Although the salt accumulation in the upper soil layers was slightly higher than the water table at 80 and 110 cm in the treatments where the water table was kept at a depth of 40 and 60 cm, there was no significant difference observed between them. Although the salt accumulation in the upper soil layers was slightly higher than the water table at 80 and 110 cm in the treatments where the water table was kept at a depth of 40 and 60 cm, there was no significant difference observed between them. On the other hand, when we consider all trial treatments, the salt concentration of the top layer increased significantly at the end of the trial (p=0.025). In comparison to the baseline, salt accumulation at depths of 40-70 and 70-110 cm is insignificant (p=0.326, p=0.715). It was determined that there was a significant difference (p=0.006-0.04) between the salt load of the incoming water and the salt load of the drained water, while there was no difference between the effects of the water table levels on the salt load of the drainage water (Table 4). 3.2. Change in soil salinity and sodium On the other hand, when we consider all trial treatments, the salt concentration of the top layer increased significantly at the end of the trial (p=0.025). In comparison to the baseline, salt accumulation at depths of 40-70 and 70-110 cm is insignificant (p=0.326, p=0.715). trial (p 0.025). In comparison to the baseline, salt accumulation at depths of 40 70 and 70 110 cm is insignificant (p 0.326, p 0.715). It was determined that there was a significant difference (p=0.006-0.04) between the salt load of the incoming water and the salt load of the drained water, while there was no difference between the effects of the water table levels on the salt load of the drainage water (Table 4). Table 4- The salinity of entering and drainage water Water table depth, cm March April May June July March April May June July Inlet water, dS m-1 3.3 3.6 4.4 4.1 4.0 3.6 3.6 3.1 4.4 4.4 Drainage water (1st year) Drainage water (2st year) 40 5.2 4.3 5.1 4.2 4.0 5.1 5.2 6.0 8.5 5.2 60 6.3 6.1 6.5 6.7 5.5 4.7 8.1 5.4 4.8 80 4.1 3.6 3.6 4.1 4.1 6.8 6.4 6.7 6.6 4.5 110 10.2 10.2 5.6 5.6 7.50 . . . . As shown in Table 5, before the experiment, the soil exchangeable sodium percentage (ESP) was 24% in the upper layer and decreased to 4% at 110 cm in depth. At the end of the trial, the ESP values in the topsoil increased significantly in all treatments compared to the initial values and reached 50-60%. In plots with a water table level of 80-110 cm, the ESP values are closer to each other in the entire profile. Crop varieties have not been shown affect ESP change. As shown in Table 5, before the experiment, the soil exchangeable sodium percentage (ESP) was 24% in the upper layer and decreased to 4% at 110 cm in depth. At the end of the trial, the ESP values in the topsoil increased significantly in all treatments compared to the initial values and reached 50-60%. In plots with a water table level of 80-110 cm, the ESP values are closer to each other in the entire profile. Crop varieties have not been shown affect ESP change. 3.2. Change in soil salinity and sodium As shown in Table 5, before the experiment, the soil exchangeable sodium percentage (ESP) was 24% in the upper layer and decreased to 4% at 110 cm in depth. At the end of the trial, the ESP values in the topsoil increased significantly in all treatments compared to the initial values and reached 50-60%. In plots with a water table level of 80-110 cm, the ESP values are closer to each other in the entire profile. Crop varieties have not been shown affect ESP change. 551 Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 Table 5- ESP status before and after trial according to groundwater depths Groundwater depth, cm Soil depth L. sitrictus P. distans F. elatior Soil depth Groundwater depth, cm L. sitrictus P. distans F. elatior ESP status before trial, (%) Before trial 0-40 24 24 24 40-60 21 21 21 60-80 13 13 13 80-110 4 4 4 ESP status after trial, (%) 40 0-40 68 61 51 0-40 39 94 42 40-60 22 20 81 80 40-60 61 53 35 60-80 16 12 25 60-80 24 74 36 80-110 13 14 16 80-110 18 29 33 60 0-40 55 50 68 0-40 53 60 31 40-60 38 12 15 110 40-60 39 53 56 60-80 28 9 43 60-80 28 21 50 80-110 21 15 17 80-110 26 23 24 Since there was no difference between the ESP values according to the crops (p=0.970ns-0.470ns), the average values are taken and the changes in ESP according to the depths of the water table and soil layers are shown in Table 5 and Figure 5. Table 5- ESP status before and after trial according to groundwater depths Since there was no difference between the ESP values according to the crops (p=0.970ns-0.470ns), the average values are taken and the changes in ESP according to the depths of the water table and soil layers are shown in Table 5 and Figure 5. 3.2. Change in soil salinity and sodium Since there was no difference between the ESP values according to the crops (p=0.970ns-0.470ns), t changes in ESP according to the depths of the water table and soil layers are shown in Table 5 and Whether there was a difference between the ESP values of the soils before the trial and the ESP values at the end of the trial was determined in the form of pairwise comparisons, taking into account the water table levels. A statistically significant difference (p<0.01; (0.000-0.007) was found between the initial ESP values of all soil layers and the ESP values at the end of the experiment at different water table levels. However, the effect of different water table levels on ESP in soil layers was found to be statistically insignificant (p>0.05; 0.95-0.719ns). Figure 4- Change of soil salinity at different water table depths Figure 4- Change of soil salinity at different water table depths 552 Bahçeci et al. - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 Figure 5- ESP status before and after trial according to different groundwater depths i Figure 5- ESP status before and after trial according to different groundwater depths 4. Conclusion The high groundwater level significantly increased the yield of pasture crops used in the experiment. Significant differences have been found between salt concentrations before and after the experiment in the trial soils. On the other hand, no difference was found between the effects of water table levels on salt and sodium accumulation in the soil. Even if the high-water table increases the grass yield in the artificial pastures planned to be created, salt and sodium accumulation should be controlled for a sustainable yield. In such areas, it seems possible to create sustainable pastures if managed drainage and groundwater are provided as leaching water at the end of the season and salt-resistant forage crops are grown. In arid and semi-arid areas under the influence of shallow groundwater for many years, uncontrolled lowering of the water table will accelerate the oxidation of organic matter as well as cause a loss of yield in these pastures. The irrigation and drainage systems in these regions should therefore be planned together, and the drainage systems to be built should be controlled drainage rather than free- flowing. Data availability: Data are available on request due to privacy or other restrictions. Data availability: Data are available on request due to privacy or other restrictions. Authorship Contributions: Concept: İ.B., Design: İ.B., Data Collection or Processing: İ.B., Analysis or Interpretation: B.B., A.F.T., Literature Search: B.B., A.F.T., Writing: B.B., A.F.T. Authorship Contributions: Concept: İ.B., Design: İ.B., Data Collection or Processing: İ.B., Analysis or Interpretation: B.B., A.F.T., Literature Search: B.B., A.F.T., Writing: B.B., A.F.T. Conflict of Interest: No conflict of interest was declared by the authors. Conflict of Interest: No conflict of interest was declared by the authors. Conflict of Interest: No conflict of interest was declared by the authors. Financial Disclosure: The authors declared that this study received no financial support. Bennett S J & Barrett-Lennard E G (2013). Predictions of groundwater depth and salinity levels land capability Crop and Pasture Science 64(3): 285-294. doi.org/10.1071/cp12417 References Acar R, Koç N, Çelik S A & Direk M (2016). The Some Grasses Forage Crops Grown in Arid Rangeland of the Central Anatolian and Properties of These Plants. 3rd International Conference on Sustainable Agriculture and Environment (3rd ICSAE) 26-28. Akhazari D, Sepehry A, Pessarakli M & Barani H (2012). Studying the Effects of Salinity Stress on the Growth of Various Halophytic Plant Species (Agropyron elongatum, Kochia prostrata and Puccinellia distans). World Applied Sciences Journal 16(7): 998-1003. Akhazari D, Sepehry A, Pessarakli M & Barani H (2012). Studying the Effects of Salinity Stress on the Growth of Various Halophytic Plant Species (Agropyron elongatum, Kochia prostrata and Puccinellia distans). World Applied Sciences Journal 16(7): 998-1003. Anonymous (2018). Republic of Turkey Ministry of Agriculture and Forestry. General Directorate of Meteorology https://www.mgm.gov.tr/index. aspx Anonymous (2018). Republic of Turkey Ministry of Agriculture and Forestry. General Directorate of Meteorology https://www.mgm.gov.tr/index. aspx Anonymous (2018). Republic of Turkey Ministry of Agriculture and Forestry. General Directorate of M aspx Ashkan A & Jalal M (2013). Effects of Salinity Stress on seed germination and seedling vigor indices of two Halophytic Plant Species (Agropyron elongatum and A. pectiniforme). International Journal of Agriculture and Crop Sciences (IJACS) 5: 2669-2676. Ashkan A & Jalal M (2013). Effects of Salinity Stress on seed germination and seedling vigor indices of two Halophytic Plant Species (Agropyron elongatum and A. pectiniforme). International Journal of Agriculture and Crop Sciences (IJACS) 5: 2669-2676. Ashour N I, Serag M S & Abd El-Haleem A K (1997). Forage production from three grass species under saline irrigation in Egypt. Journal of Arid Environments 37(2): 299-307. doi.org/10.1006/jare.1997.0284 Ashour N I, Serag M S & Abd El-Haleem A K (1997). Forage production from three grass species under saline irrigation in Egypt. Journal of Arid Environments 37(2): 299-307. doi.org/10.1006/jare.1997.0284 Ashour N I, Serag M S & Abd El-Haleem A K (1997). Forage production from three grass species under sal Environments 37(2): 299-307. doi.org/10.1006/jare.1997.0284 Atalay M & Secerli L (1971). Konya arazi tasnif raporu, Topraksu Bölge Müdürlüğü, Konya, (In Turkish) Bennett S J & Barrett-Lennard E G (2013). Predictions of groundwater depth and salinity levels land capability assessment using site indicator species. Crop and Pasture Science 64(3): 285-294. doi.org/10.1071/cp12417 Bennett S J & Barrett-Lennard E G (2013). Predictions of groundwater depth and salinity levels land capability assessment using site indicator species. Crop and Pasture Science 64(3): 285-294. doi.org/10.1071/cp12417 553 Bahçeci et al. References - Journal of Agricultural Sciences (Tarim Bilimleri Dergisi), 2023, 29 (2): 546-554 Brotherson J D (1987). 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hal-02081353-PlosOne_NaudinotEtAl_2017.txt_2	French-Science-Pile	Open Science	Various open science	2,019	Divergence in the evolution of Paleolithic symbolic and technological systems : The shining bull and engraved tablets of Rocher de l'Impératrice. PLoS ONE, 2017, 12 (3), pp.e0173037. ⟨10.1371/journal.pone.0173037⟩. ⟨hal-02081353⟩	None	English	Spoken	5,490	10,500	In addition, the visual impact of the figure is enhanced by the coloring of the tracings with black pigment revealed by the cleansing of the incisions and evidenced by Raman spectrometry. Raman measurements were performed in the engravings at the black colorations and all recorded Raman spectra underline the presence of amorphous carbon (commonly called as a PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 16 / 26 The shining bull: A missing link for Paleolithic art pigment “black carbon”). Even if black carbon is by nature poorly organized regarding its atomic structure, the Raman analyses could reveal a clear and specific structural signature (S1A Fig). Such result clearly excludes other possible black pigments such as manganese oxides which should be detected by Raman spectroscopy. However, the tablet is made of shale and such rocks are known to contain some carbonaceous materials [41]. Spectra were then recorded on the rock outside of the engravings and without macroscopically or microscopically noticeable black coloration; underlining that some amorphous carbon can be detected in the shale itself as illustrated in S1A Fig. However, by overlapping all the spectra from the pigment and the shale some slight differences could be observed in the spectral shape (S1B Fig). Even if it appears tiny, such differences are related to structural variations of the carbonaceous material according to their origin (carbonization or rock diagenetic process) [42], allowing the differentiation of the carbonaceous matter from various origins. The carbon signal detected in the engraving, therefore, does not originate from the shale itself. The carbon can only be explained by people introducing carbon onto the tablet, which is preserved in the engravings. Such enhancement of engraved motifs with color is rather unusual even if documented from time to time (some mobile art pieces from La Vache, Gourdan or Enlène for instance; [43–45]). A very similar pattern of large and deep rays is found on another small fragment (Fig 5: fragments 167–168) suggesting that other shining figures could have been depicted. The “shining bull” may not be alone. Besides this special visual valorization, the two animals exhibit a naturalistic rendering. Their outlines are shapely (convexity of the eye-socket, the chin and the lower jaw). The horns are sinuous. The tracings which diverge to their end would rather indicate two horns in single linear tracing seen in three-quarters. On side B, the two parallel tracings seem to belong to the same horn directed upward. The sensory organs are indicated but not as precisely drawn: round eye, mouth line. A nostril and a short pointed ear directly stuck in the outline at the back of the horn complete the aurochs of tablet 317B. The two subjects show some variations: the shaping work is more sensitive on the aurochs of 317B, and is also more detailed; the perspective applied to the horns also seems to be different. On the aurochs of 317A, a special attempt to reproduce the volume of the animal was made: the natural foliation of the shale was used and slightly retouched with scratching to suggest the relief of the zygomatic arch. The coat was given particular attention, reproduced in different ways: a series of parallel oblique short rectilinear lines stuck to the back line, a grid (317A) or patches of vertical and long superficial fine incisions at the base of the neck. Using 317A as a guide, other pieces could be fragments of animal silhouettes and/or internal fillings (Fig 5: fragments 46, 164, 46, 673, TAM. L12-E-CA101). Side B of 317 also bears numerous fine incisions of a former set made of animal depictions (herbivore forequarters?) covered in what currently appears as “unorganized tracings.” 4. Discussion This collection testifies to formal and technical homogeneity, with some particular graphic conventions shared by different figures: the angular and pinched muzzles of the little horse on slab 741B and the horse on fragment 607, and the representation of the coat with a series of short oblique straight lines along the outlines of the horses on slab 741 and the aurochs on slab 317. Because of the obvious stylistic unity of these figures, we consider the corpus as a coherent assemblage in which coexist two formal renderings of the animal, one naturalistic, the other schematic. Even if most of these artifacts were discovered in the disturbed unit of the shelter, the entire corpus of engraved tablets can reasonably be attributed to the Early Azilian PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 17 / 26 The shining bull: A missing link for Paleolithic art occupation(s) of Le Rocher de l’Impératrice as no evidence of occupation prior to the Azilian was found on the site. The 45 engraved stone fragments from Le Rocher de l’Impératrice are the oldest graphic remains in Brittany and prove to be exceptional for the EA. In France, less than a dozen sites have yielded decorated items related to this techno-complex, most of them from uncertain contexts. Among them, only three collections of mobile art can be clearly attributed to this culture: the 82 engraved lithic artifacts of Murat rock-shelter (layers B and D of Lemozi, layer IV of Lorblanchet: [25]), the five flakes with engraved cortex of Le Closeau [14], an engraved pebble of Bois-Ragot (layer 4: [26]). A single parietal art site is attributed to the EA- Gouy cave [23, 46]–on the basis of the lithic industry found at the site; however, no direct link exists so far between the parietal set and the archaeological deposit. In this regard, the collection of Le Rocher de l’Impératrice considerably increases the data. Even if it is difficult to draw general conclusions about the Early Azilian symbolic system from these few sites, and the very restricted corpus for Le Closeau and Le Bois-Ragot, a coherent image of these graphic works still appears: the notable selection of small pebbles and slabs as blanks, and the mostly unifacial sets made of a unique motif or a very small number of superimposed motifs. The iconography combines both geometric themes such as checkered triangles, lines, or beams, and figurations with bestiaries dominated by horses and aurochs (Fig 8). The schematic animals depicted in Gouy cave bear stylized coats in the form of grid patterns or parallel lines reminiscent of the aurochs on tablet 317 [46], together with angular and pinched muzzles for the horses. Murat rock-shelter figures also show two types of formal rendering: schematic representations cohabit with realistically shaped and dynamic animals with simplified internal details (Fig 8). The EA iconography of Le Rocher de l’Impératrice is very similar to the Upper-Final Magdalenian and several elements suggest direct continuity. That is the case of the selected materials to be engraved: pebbles and tablets are also abundantly used in large lithic mobile art assemblages such as Limeuil, Gourdan, Gönnersdorf, La Peña de Estebanvela and Foz do Metal [40, 48, 50–53]. This similarity can also be seen in the technical preference for engraving instead of painting. The compositions associating geometric patterns, figures and unorganized tracings is another common pattern. The themes are also very similar: the geometric register appears to remain rather unchanged with the predominance of chevrons, grids, lines beams, together with triangles and ellipses. As to the figurative register, aurochs and horses also fill the Upper/Final Magdalenian bestiary even if aurochs are not very common. The formal conventions of the animals in Le Rocher de l’Impératrice match well with those documented of the Upper-Final Magdalenian: a realistic rendering with complete bodies, with the two finely reproduced pairs of legs, seen in real perspective, shapely silhouettes, and numerous anatomical details depicting the sensorial organs, the internal volumes and the coat. Macrocephaly of the horses is another strong figurative trend [5, 54]. These formal traits are shared within the whole graphic production of the second half of the Magdalenian in Western Europe, from the Paris Basin with the engraved pebble from Étiolles [55] to the North of Spain (Abauntz: [56]; in others). They are also the ones of some parietal paintings directly dated between 13,500 and 12,500 BP at Niaux [57], La Pasiega C [58], and El Castillo [58]. Some particular MiddleUpper Magdalenian conventions can be more specifically pointed: the depiction of the zygomatic depression (aurochs 317A, horse 741A?), the stylized representation of the coat as a series of short, straight oblique lines stuck to the back line (aurochs on fragment 317, horse 741A) or as a fine hatching filling inside the legs and the belly (horses on tablet 741) [5, 44, 48– 49, 51, 59]. A less detailed formal rendering could be argued for the engravings at Le Rocher de l’Impératrice but simplified representations are also found in the Upper-Final Magdalenian together with very naturalistic figures, some turning very schematic, sometimes within the PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 18 / 26 The shining bull: A missing link for Paleolithic art Fig 8. doi:10.1371/journal.pone.0173037.g008 PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 19 / 26 The shining bull: A missing link for Paleolithic art same assemblage (e.g. abri Mège, La Mairie, Le Morin, Limeuil, Laugerie-Basse, Rochereil or La Madeleine–[49, 60]). From these results we suggest continuity in the iconographic tradition between the Upper Magdalenian and the EA, as already suggested by M. Lorblanchet in Murat [25]. The dearth of artistic items from the EA nevertheless limits our estimation of the degree of longevity after the Upper Magdalenian. However, the special valorization of the aurochs as shown by the “shining bull” could indicate some symbolic recoding of the traditional bestiary. Evidence is too scarce at the moment to develop this hypothesis further. We also suggest that the chronocultural attribution of some Upper Magdalenian mobile art collections should be reevaluated with the possibility that some may be more recent since many assemblages were actually collected in sites where the Upper Magdalenian layers are generally covered with EA and LA layers (e.g., La Madeleine, Villepin, grotte Richard; [48]; Laugerie-Basse: [61]). However, this continuity is particularly striking with regard to the Southern cultural context of Northern Spain where the Upper-Final Magdalenian lasts until 11,800–11,600 BP whereas the EA develops in France [62]. Thus, two neighboring cultural groups with different domestic and hunting equipment but a common iconography seem to coexist (i.e., in others La Vache, Le Portel, Ekain VIb, Oscura de Ania IIIa, Abauntz 2r, Las Monedas, La Pasiega C–[44, 63–66]). These results provide the first picture of a graphic tradition in the EA clearly distinctive from the LA whose iconography is dominated by painted and engraved abstract patterns on pebbles; however such abstract pebbles could be already present in the EA in Murat rock-shelter (layers B and D of Lemozi, layer IV of Lorblanchet). Similar pebbles in Bois-Ragot (layer 4), Gay rock-shelter and Rochedane [2, 67], are likely from mixed contexts containing EA and LA materials. Where abstract pebbles are found in secure contexts, there is a clear iconographic schism between the EA and LA in regard to 1) a shift in material selection with use only of standardized pebbles, 2) painting as a preferred technique, 3) the explosion of abstract expressions in the shape of repeated simple patterns (points, lines, crosses, chevrons), 4) the correlative scarcity of depiction whether animal or human, and 5) a change in the formal rendering of the figures in favor of geometric schematism. The lines are simplified and rugged (triangular legs without hoof), the bodies are wide and the heads atrophied. Most often, the internal details are missing. The bodies can be blank or filled with patches of long tracings, oblique or horizontal on the flank, vertical in the legs (Murat: [25]). Similar iconographic changes accompany the spread of the classical Azilian in the North of the Iberian peninsula [53, 68]. The data presented here highlight a discrepancy between technical and symbolic changes during the azilianization process. If the graphic corpus of Le Rocher de l’Impératrice testifies to iconographic continuity with the Magdalenian, the lithic assemblage already bears all the Azilian characteristics with especially the exclusive use of soft hammer stones and the exclusive use of axial projectile points to the detriment of backed bladelets. The major transformation of the symbolic system with the development of a geometric expression and the correlative nearabandonment of animal and human figures seems to start later. While the data are still too scarce to determine precisely when a shift in symbolic system begins, those available data suggest the beginning of the Allerød. The move away from Magdalenian and EA iconography probably occurred after the beginning of the LA. This timing is suggested by the discovery of a horse head still bearing the EA formal conventions (arched nose, pinched lip, few details but the mention of the zygomatic depression and the coat represented by series of long vertical lines) engraved on cortex found in layer III.20 at Pincevent dated around 13,600 cal. BP at the beginning of the LA ([69–70]; Fig 9). This study underlines a difference in rhythm between technological and symbolic changes of the Magdalenian and EA. Our data suggest a progressive dissolution of the Magdalenian lithic standards while conventions of Magdalenian iconography seem to change later and PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 20 / 26 The shining bull: A missing link for Paleolithic art Fig 9. Similarities of conventions in the formal rendering of the horse heads from Le Rocher de l’Impératrice and Pincevent (layer III.20) (photo N. Naudinot; tracing D. Baffier in [69]). doi:10.1371/journal.pone.0173037.g009 abruptly by the Allerød. The likely disappearance of animal and human figures in favor of an exclusive abstract expression is a crucial iconographic shift with regards to the other former Upper Palaeolithic societies and to the subsequent final Paleolithic populations in Western Europe. While it is possible that this picture of a clear scission in the symbolic sphere is only caused by the lack of data between EA and LA, the findings at Le Rocher de l’Impératrice suggest that Magdalenian iconography is greatly extend in time. The onset of the Azilian is not a package of changes affecting both the symbolic system and technical equipment. It is essential to understand the origin of this critical evolution in symbolic expression, which may bear important ideological and sociological significance during the Azilian. The simple techniques, motifs, as well as the large residential sites in which the decorated pebbles are generally unearthed would rather indicate common productions of these items in the LA. This marks an important shift from Magdalenian graphic production [62], which emphasizes a complex technical savoir-faire, realism, and a degree of detail in numerous representations that are without any doubt the work of specialists who went through a distinct learning process [71– 72]. According to Gonzalez Morales [73], the abandonment of figures for geometric expression would be linked to changes in the techniques used and in the actors, and the three would result from a complete series of global changes in the social organization of graphic production. In parallel, the decrease in savoir-faire can also be identified in the lithic and bone technology of the LA. It is tempting to see these phenomena as a possible development of a common production strategy that minimized the importance of specialists in both graphic and technical activities. These data could suggest a major sociological shift during the Allerød. Changes in mobility patterns, judged by the few data available, suggest a potential increase in residential mobility during the LA [6, 13, 74–76]. This change in mobility is important to consider as the reorganization of activities in time and space may be profoundly linked with these cultural changes. In human cultural evolution, changes in the cultural system do not necessarily affect all elements of material culture at the same time [62, 77]. This is sometimes contrary to the general idea conveyed in archaeological studies. This vision is mostly perpetuated by the nature of the archaeological record as our theories and interpretations are often based on an extremely fragmentary subset of remains from past cultural systems. Sites like Le Rocher de l’Impératrice PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 21 / 26 The shining bull: A missing link for Paleolithic art with remains of various parts of the cultural system allow us to discuss and compare differences in the tempo of cultural change. In this case, the development of Azilian cultural or symbolic acts appear to lag behind major technological adaptations. Supporting information S1 Table. Engraved tablets database. Captions: L/D: Large and deep engraving; F/S: Fine and superficial engraving; F.D: Fine and deep engraving; W/D: Wide and deep engraving; W/D +Ch: Wide and very deep engraving with champlevé; SPSC: Small patches of sediment crusts; SPSD: Small patches of sediment deposit; SPSDC: Small patches of sediment deposit and crusts; HP: Highly patinated surface; HEE: Highly eroded edges; HPT: Highly patinated tracing; OAA: Other anthropological action; 1: Blue grey; 2: Visible lamination; 3: metallic light grey; 4: light and dark blue; 5: Shiny lamination; 6: Pearlescent pinkish light grey; 7: Matt slate grey; 8: Matt dark blue grey; 9: Grey (DOCX) S1 Fig. a)Several Raman spectrum obtained on the black pigment in the engraving compared with some obtained on the shale tablet out of engraved areas and without visible black colour. These signatures are constituted by the symmetric stretching of carbon-carbon single bonds (band around 1367 cm-1) and carbon-carbon double bonds (band around 1597 cm-1). Spectra are baseline corrected to remove the fluorescence contribution. Several spectra from the two kind of areas are overlapped to underline reproducibility, and the two vertically shifted for readability. b) Same spectra from both areas overlapped in order to highlight the shapes different between the pigment and the carbonaceous matter from the shale. (DOCX) Acknowledgments This work was conducted with the help of the SRA Bretagne (French Ministry of Culture and Communication), the Conseil départemental du Finistère and the municipality of PlougastelDaoulas. We would like to also thank all the students and volunteers involved in the excavation of Le Rocher de l’Impératrice. We are extremely thankful to Robert L. Kelly (University of Wyoming) and Joseph A.M. Gingerich (Ohio University and The Smithsonian Institution) who helped considerably with the manuscript and François Bon (University of Toulouse II) for his precious advice. Thank you to Marco Peresani (Ferrare University), Patrick Paillet (UMR 7194, Muséum national d’histoire naturelle), Ludovic Mevel (UMR 7041, Arscan) and the other three anonymous reviewers of this paper. Author Contributions Conceptualization: NN CB. Data curation: NN. Formal analysis: NN CB ML SB CP LBG ITP. Funding acquisition: NN. Investigation: NN MLG. Methodology: NN CB ML SB CP LBG ITP. Project administration: NN. PLOS ONE \| DOI:10.1371/journal.pone.0173037 March 3, 2017 22 / 26 The shining bull: A missing link for Paleolithic art Resources: NN MLG. Supervision: NN. Visualization: NN CB ML. Writing – original draft: NN CB ML CP LBG. Writing – review & editing: NN CB. References 1. Piette E. L’art pendant l’âge du renne. Paris: Masson; 1907. 2. Thévenin A. 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Bulletin de la Société Préhistorique Française, 1992; 89, 9: 270–274. 58. Moure Romanillo J. A. and González Sainz C. Cronologı́a del arte paleolı́tico cantábrico: últimas aportaciones y estado actual de la cuestión. In: Actas do 3e Congresso de Arqueologia Peninsular (Vila Real, 1999). Vol. II: Paleolı́tico da Penı́nsula Ibérica. Porto: ADECAP; 2000. pp.461-473. 59. Man-Estier E. & Paillet P. Rochereil et l’art magdalénien de la fin du Tardiglaciaire dans le nord du Périgord. In: Groenen M, editor. Aesthetic expressions and technical behaviours in the Palaeolithic Age (Proceedings of the XVI IUPPS World Congress, BAR International Series 2496). Oxford: Archeopress; 2013. pp 7–36. 60. Roussot A. Art mobilier et art pariétal du Périgord et de la Gironde: comparaisons stylistiques. In: Clottes J, editor. L’art des objets au Paléolithique, 1, l’art mobilier et son contexte, colloque international de Foix-le Mas-d’Azil. Paris: Ministère de la Culture; 1987, pp. 189–205. 61. 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https://openalex.org/W1874784862	OpenAlex	Open Science	CC-By	2,015	Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis	Chunyan Bao	English	Spoken	9,595	16,921	MIT Open Access Articles Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation: Bao, Chenchen, Joao Conde, James Curtin, Natalie Artzi, Furong Tian, and Daxiang Cui. “Bioresponsive Antisense DNA Gold Nanobeacons as a Hybrid in Vivo Theranostics Platform for the Inhibition of Cancer Cells and Metastasis.” Scientific Reports 5 (July 20, 2015): 12297. As Published: http://dx.doi.org/10.1038/srep12297 MIT Open Access Articles Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation: Bao, Chenchen, Joao Conde, James Curtin, Natalie Artzi, Furong Tian, and Daxiang Cui. “Bioresponsive Antisense DNA Gold Nanobeacons as a Hybrid in Vivo Theranostics Platform for the Inhibition of Cancer Cells and Metastasis.” Scientific Reports 5 (July 20, 2015): 12297. As Published: http://dx.doi.org/10.1038/srep12297 P bli h N t P bli hi G MIT Open Access Articles The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation: Bao, Chenchen, Joao Conde, James Curtin, Natalie Artzi, Furong Tian, and Daxiang Cui. “Bioresponsive Antisense DNA Gold Nanobeacons as a Hybrid in Vivo Theranostics Platform for the Inhibition of Cancer Cells and Metastasis.” Scientific Reports 5 (July 20, 2015): 12297. Citation: Bao, Chenchen, Joao Conde, James Curtin, Natalie Artzi, Furong Tian, and Daxiang Cui. “Bioresponsive Antisense DNA Gold Nanobeacons as a Hybrid in Vivo Theranostics Platform for the Inhibition of Cancer Cells and Metastasis.” Scientific Reports 5 (July 20, 2015): 12297. Version: Final published version: final published article, as it appeared in a journal, conference proceedings, or other formally published context Terms of use: Creative Commons Attribution Terms of use: Creative Commons Attribution www.nature.com/scientificreports www.nature.com/scientificreports Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis received: 01 April 2015 accepted: 23 June 2015 Published: 20 July 2015 Chenchen Bao1,, João Conde2,3,, James Curtin4, Natalie Artzi2,5, Furong Tian6 & Daxiang Cui1 Chenchen Bao1,, João Conde2,3,, James Curtin4, Natalie Artzi2,5, Furong Tian6 & Daxiang Cui1 Gold nanobeacons can be used as a powerful tool for cancer theranostics. Here, we proposed a nanomaterial platform based on gold nanobeacons to detect, target and inhibit the expression of a mutant Kras gene in an in vivo murine gastric cancer model. The conjugation of fluorescently-labeled antisense DNA hairpin oligonucleotides to the surface of gold nanoparticles enables using their localized surface plasmon resonance properties to directly track the delivery to the primary gastric tumor and to lung metastatic sites. The fluorescently labeled nanobeacons reports on the interaction with the target as the fluorescent Cy3 signal is quenched by the gold nanoparticle and only emit light following conjugation to the Kras target owing to reorganization and opening of the nanobeacons, thus increasing the distance between the dye and the quencher. The systemic administration of the anti-Kras nanobeacons resulted in approximately 60% tumor size reduction and a 90% reduction in tumor vascularization. More important, the inhibition of the Kras gene expression in gastric tumors prevents the occurrence of metastasis to lung (80% reduction), increasing mice survival in more than 85%. Our developed platform can be easily adjusted to hybridize with any specific target and provide facile diagnosis and treatment for neoplastic diseases. Amolecular beacon is a hairpin DNA single-stranded oligonucleotide, which transports a dye and a quencher at both ends. In the absence of a complementary target, the stem-loop structure is closed imposing the dye and the quencher to a close proximity, causing a fluorescence quenching. In the pres- ence of a complementary target, the stem-loop sequence is opened and the molecular beacon hybridizes, 1Institute of Nano Biomedicine and Engineering, Key Lab. of Thin Film and Microfabrication Technology of Ministry of Education, Department of instrument science and engineering, School of Electronic Information and Electrical Engineering, National Center for Translational Medicine, Shanghai Jiao Tong University, P.R.China. 2Massachusetts Institute of Technology, Institute for Medical Engineering and Science, Harvard-MIT Division for Health Sciences and Technology, Cambridge, Massachusetts, USA. 3School of Engineering and Materials Science, Queen Mary University of London, London, UK. 4School of Food Science and Environmental Health, College of Sciences and Health, Dublin Institute of Technology, Cathal Brugha Street, Dublin, Ireland. www.nature.com/scientificreports/ The 5-year survival rate for patients with advanced GC is only around 10%; the rest of the patients usually have median survival times of 8–12 months26. Improving the survival of GC patients requires the development of novel diagnostic and anti-tumor therapy. A Kras mutation have recently been described in gastrointestinal stromal tumors (GISTs), yielded a high number of KIT mutant GISTs and reported also in wild type for KIT and PDGFRA27,28. yp Kras is one of the most commonly activated oncogenes in human cancer, with 17 to 25% of all human tumors harboring an activating Kras mutation29 In addition to gastrointestinal tumors, over 90% of pan- creatic adenocarcinomas, 30%–50% of colorectal cancers, 55% of thyroid cancers, 35% of lung cancers, and 35% of rhabdomyosarcomas harbor mutated RAS genes30. Previously, Conde et al. developed an in vitro gold-nanobeacon system as a proof-of-concept to follow RNA synthesis in real time in bulky solutions and for antisense DNA and RNA interference (RNAi), from gene specific silencing to silence-the-silencers in vitro only31–33. From these studies, the authors proved the potential of a single molecular nanoconjugate to intersect all RNA pathways in vitro: from gene specific downregulation (ie. inhibit a GFP reporter) to silencing siRNA and miRNA pathways31.h i Here, we used antisense DNA gold nanobeacons to selective inhibition of mutant Kras in vivo. These bioresponsive gold nanobeacons have a unique and positive effect on metastatic cell colonization in the lungs in a syngeneic and spontaneous gastric cancer mice model. This nanobeacons platform is of high therapeutic potential and serves as a theranostics system of orthotopic gastric tumors but also as anti-metastasis treatment, where nanobeacons show a strong involvement in both local and distant lesions.h Therefore, we designed and developed an in vivo universal tool based on AuNPs functionalized with a dye labeled hairpin-DNA, i.e. gold-nanobeacon to inhibit simultaneously cancer cells and metastasis in a murine tumor model following systemic administration via the antisense silencing of a mutant transcript of Kras gene (see Fig. 1A). The NPs used in this study consist of a AuNP core decorated with and a thiol-DNA-hairpin labelled with a Cy3 dye. A polyethylene glycol (PEG) is also used as a spacer between the antisense DNA hairpins, increasing the nanostructure stability in biological medium32,33 (see Supplementary Figure S1 for PEG optimization and quantification). www.nature.com/scientificreports/ creating a double-stranded 3D structure. With the opening of the stem-loop, the spatial distance between dye and quencher increases and the fluorescence is reestablished1.i creating a double-stranded 3D structure. With the opening of the stem-loop, the spatial distance between dye and quencher increases and the fluorescence is reestablished1.i l Molecular beacons can be optimized to match with specific experimental requirements. By controlling the length of both stem and loop a broad range of temperatures for target detection can be achieved, as well as lower signal-to-background ratios and/or different dissociation rate constants2. Although very productive for in vitro experiments, utilization of molecular beacons ex vivo and in vivo has not been successful, probably due to the reduced chemical stability of nucleic acids in biological media and fragile protection against serum nucleases.hi p g The growing field of nanotechnology has brought a wide variety of tools for molecular analysis and clinical imaging and theranostics with increased sensitivity and specificity3–9. Due to their opti- cal properties, nanomaterials and nanoparticles in particular have been used for nucleic acid screen- ing methodologies, specifically through thiolated DNA oligonucleotides capable of recognizing specific nucleotide sequences10–13. The combination of the optical properties of nanoparticles with the demon- strated advantages of standard molecular beacons can prove to be extremely useful in molecular biology. Gold nanoparticles (AuNPs) have been demonstrated to modulate dye emission in their vicinity14, from fluorescence quenching to fluorescence enhancement15, which suggests AuNPs as an excellent substitute of classic quenchers. Additionally, AuNPs provide the possibility for in vivo studies of DNA hairpin-like structures as they offer protection against degradation by nucleases16,17 and can act as both transfection and targeting vectors18–20 controlling crucial cellular processes such as antisense DNA and RNA inter- ference pathways. p y Antisense DNA21,22 pathways have emerged as powerful and useful tools to block gene function and for sequence-specific posttranscriptional gene silencing. The discovery of their prominent role in regu- lation of specific gene expression in numerous disease states, such as cancer resulted into an enormous effort towards the development of new efficient delivery systems that control those pathways23,24. ffi Gastric cancer (GC) is a prevalent cancer in Asia, with most new cases and deaths in China, Japan and Korea. It is the third most common malignant tumor and causes 300,000 deaths in China each year25. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis 5Department of Anesthesiology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA. 6Focas Research Institute, Dublin Institute of Technology, Camden Row, Dublin, Ireland. These authors contributed equally to this work. Correspondence and requests for materials should be addressed to J.C. (email: jdconde@mit.edu) or F.T. (email: furong.tian@dit.ie) Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 1 www.nature.com/scientificreports/ www.nature.com/scientificreports/ Under hairpin configuration, the proximity of the Cy3 dye to the AuNP (that works as quencher) leads to fluorescence quenching. Hybridization of the DNA hairpin to a complementary target (i.e. Kras mRNA) restores fluorescence emission due to the gold nanobeacons’ conformational reorganization that causes the fluorophore and the quencher to part from each other, yielding a quantitative response. These molecular beacons have the ability to interlock or hybridize with the target Kras mRNA (blocking specific translation initiation signals of Kras gene) thus inhibiting the translation of the target protein. In addition to designing anti-Kras nanobeacon that detects and inhibits Kras mRNA, a nonsense nano- beacon (that does not hybridize with any target) was developed as an internal control (Supplementary Figure S2 and Table S1). Oligomers sequences and 2-dimensional structures of the beacons at 37 °C are depicted in Supplementary Table S1. p pp y Transmission electron microscopy (TEM) images of the DNA antisense gold nanobeacons anti-Kras showed an average diameter of the gold core of 15.1 ± 1.1 nm (Supplementary Figure S3A). The final bea- con:NP ratio was around 45:1 (see Supplementary Table S2). The mean particle diameter of nanobeacons Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 2 ure.com/scientificreports/ Figure 1. (A) Gold nanoparticles (AuNPs) functionalized with a dye (Cy3) labeled hairpin-DNA, i.e. gold- nanobeacon (Au-nanobeacon), designed to detect, target and inhibit the expression of Kras gene in an in vivo murine gastric cancer model. Under hairpin configuration, proximity to gold nanoparticles leads to fluorescence quenching; hybridization to a complementary target restores fluorescence emission due to the Au-nanobeacons’ conformational reorganization that causes the dye and the AuNP to part from each other. The scheme was designed and produced on Adobe Illustrator CC 2014 by J.Conde. (B) Cy3 emission at 570 nm after hybridization with increasing amounts of complementary and non-complementary ssRNA targets (0 to 3E-05 M). (C) Flow cytometry analysis comparing nanobeacon anti-Kras treated cells to nanobeacon nonsense. (D) The silencing effect was expressed as a concentration-dependent decrease in Kras relative expression. Anti-Kras nanobeacons showed potent silencing effects in human gastric cancer cells (MGC-803) with an ED50 between 1 and 5 nM. (E) Live imaging images of anti-Kras nanobeacon (previously incubated with Kras complementary target) compared to nanonsense nanobeacon and a blank b www.nature.com/scientificreports/ Figure 1. (A) Gold nanoparticles (AuNPs) functionalized with a dye (Cy3) labeled hairpin-DNA, i.e. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 www.nature.com/scientificreports/ When the concentration of Cy3-labeled DNA oligo was fixed at 1 μ M, the increase in AuNPs concentration from 0 to 8 nM in the reaction mixture resulted in a signifi- cant increase in Cy3 fluorescence quenching (Supplementary Figure S6B), demonstrating the presence of NSET between the Cy3 groups and the AuNPs. These data showed a quenching efficiency of nearly 90% when the concentration of AuNPs reached 3 nM, whereas nearly 100% when the concentration of AuNPs reached 8 nM. Conversely, the Cy3 emission can be restored after hybridization with the complementary ssDNA Kras target. Increasing amounts of complementary target results in elevated emission form the Cy3 dye at 570 nm until reaching a plateau at 1E–05M of target (Fig. 1B).lii y y g p g g Indeed, analytical flow cytometry data confirmed the signal specificity of the nanobeacon incubated in human gastric cancer cells (MGC-803), where a substantial increase in Cy3 intensity signal is observed only for nanobeacons anti-Kras when compared to nanobeacon nonsense (Fig. 1C).fi We next assessed in vitro gene silencing efficiency in the human gastric cancer cell line MGC-803 of both anti-Kras and nonsense nanobeacons. The ability of both anti-Kras and nonsense nanobeacons to silence Kras was evaluated in gastric cancer cells, where only the anti-Kras nanobeacons provided a robust knockdown in a dose dependent manner, with a median effective dose (ED50) between 1 and 5 nM. Specifically, more than 80% silencing could be observed at the mRNA level by qPCR at a dose of 50 nM at 48 hours of incubation (Fig. 1D).ll In order to corroborate flow cytometry data (Fig. 1C) the fluorescence signal from gold nanobea- cons previously incubated with a ssRNA Kras target (mimicking Kras mRNA) was evaluated by the live imaging system (Fig. 1E). As expected, only anti-Kras nanobeacons produce a significant increase in fluorescence signal, when compared to nonsense nanobeacons and a blank.hfi l The efficiency of the anti-Kras nanobeacon probes in sensing and in silencing gastric cancer cells in vivo was evaluated in an orthotopic gastric cancer mice model. Human gastric cancer cell line MGC-803 was used to generate the orthotopic tumor model in nude mice. After the formation of the tumor tissue, the tumor tissue stability in nude mice was passaged three more generations in subcutaneous tumors, before transferred into nude stomach via small animal surgery. www.nature.com/scientificreports/ gold- nanobeacon (Au-nanobeacon), designed to detect, target and inhibit the expression of Kras gene in an in vivo murine gastric cancer model. Under hairpin configuration, proximity to gold nanoparticles leads to fluorescence quenching; hybridization to a complementary target restores fluorescence emission due to the Au-nanobeacons’ conformational reorganization that causes the dye and the AuNP to part from each other. The scheme was designed and produced on Adobe Illustrator CC 2014 by J.Conde. (B) Cy3 emission at 570 nm after hybridization with increasing amounts of complementary and non-complementary ssRNA targets (0 to 3E-05 M). (C) Flow cytometry analysis comparing nanobeacon anti-Kras treated cells to nanobeacon nonsense. (D) The silencing effect was expressed as a concentration-dependent decrease in Kras relative expression. Anti-Kras nanobeacons showed potent silencing effects in human gastric cancer cells (MGC-803) with an ED50 between 1 and 5 nM. (E) Live imaging images of anti-Kras nanobeacon (previously incubated with Kras complementary target) compared to nanonsense nanobeacon and a blank tube. anti-Kras is 25.9 (±3.5) nm and nanobeacon nonsense is 24.7 (±2.9) nm as measured by DLS and with an SPR peak at 525 nm as evaluated by UV-Vis extinction profiles (Supplementary Figure S3B), and slightly anionic (−25.6±1.2 mV) via zeta-potential (Supplementary Table S2). Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 3 www.nature.com/scientificreports/ The stability of gold nanobeacons towards increasing concentrations of Glutathione (GSH) and DNase was also evaluated (Supplementary Figure S4A,B). These data confirms the stability of the gold nano- beacons to intracellular concentrations of GSH (Supplementary Figure S4A) and DNase (Supplementary Figure S4B). g ) The fluorescent emission from the Cy3 (Emi = 570 nm) is efficiently quenched by the ~14 nm AuNP due to a nanosurface energy transfer (NSET) effect34. The fluorescence is “OFF” when the hairpin-DNA-Cy3 is closed (i.e. no hybridization with the complementary target). The fluorescence is “ON” when the hairpin-DNA-Cy3 hybridizes with a Kras complementary target and an increase in Cy3 emission occurs (Supplementary Figure S5). It is worth noting that the fluorescence emission intensity of Cy3-labeled DNA oligo is significantly decreased after the reaction with gold nanoparticles, due to the quenching effect between the Cy3 dye and the gold surface. This effect occurs once a good overlap between dye emission and quencher (AuNP) absorbance is observed, providing an indication of the quenching effi- ciency (Supplementary Figure S6A). www.nature.com/scientificreports/ An OB adhesive construction of ortho- topic gastric tumor transplantation was applied for tumor tissue adhesion. OB glue paste technique is commonly used to establish nude mouse human gastric cancer orthotopic transplantation models. The OB glue paste technique is easy to perform and the biological behaviors of the nude mice human gas- tric cancer orthotopic transplantation models established with this technique are similar to the natural processes of growth and metastasis of human gastric cancer35. Four weeks after the OB surgery and the establishment of the orthotopic model, mice were tail-vein injected with gold nanobeacons and in vivo imaging was used to track simultaneously organs biodistribution, tumor size and nanobeacon probes signal after hybridization to the target monitored by fluorescence emission over a period of 120 hours (5 days) following nanobeacons injection (Fig. 2). The trial endpoint was 5 days once after that the signal from the nanobeacons decreased dramatically. In vivo imaging of mice administered with 5 or 50 nM of anti-Kras nanobeacons or nonsense nanobeacons (50 nM) revealed that only nanobeacons anti-Kras were able to promote efficient and sustained detection of primary gastric tumor (Fig. 2A,B), with an approximately 60% tumor size reduction 5 days after intravenous injection, when compared to nonsense nanobeacon. Concerning nanobeacon probes imaging, fluorescence images of treated mice revealed, as expected, that fluorescence signal is OFF at day 0 (0.5 hours after tail-vein injection) and is turned ON 4 hours later, and maintains the signal until 120 hours (Fig. 2A), only for the anti-Kras nanobeacons and not for the nonsense nanobeacons. Recently, a similar system using gold nanobeacons for both gene and drug delivery in a multid- rug resistance mice model was described36. The authors reported that gold nanobeacons could enter cells probably via an endocythosis-mediated mechanism. Confocal images of breast cancer cells further revealed that the gold nanobeacons were colocalized in intracellular organelles such as endosomes and/ or lysosomes at 24 hours and could escape out of this organelles at 48 hours. If the majority of gold nano- beacons escape lysosome the mechanism behind the cellular uptake and endosomal escape is probably an endocytosis mechanism mediated by caveolae receptors, once the escape from the endosomal com- partments happens before lysosomal activation. This mechanism represents an alternative pathway with distinct cellular compartments to avoid lysosomal degradation. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 www.nature.com/scientificreports/ The anionic nature of the nanobeacons may be endocytosed through the interaction with the positive site of the proteins in membrane, and they Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 4 ure.com/scientificreports/ Figure 2. (A) Epi-fluorescence analysis of nude mice bearing orthotopic human gastric cancer cells, 0.5, 4, 24, 72 and 120 hours after tail-vein injection of nonsense (50 nM) and anti-Kras (5 and 50 nM) nanobeacons. Arrows represent gastric tumors. (B) Epi-fluorescence images of whole body organs to evaluate selective signal from the gold nanobeacons accumulated in tumors. (C) Quantification of anti-Kras nanobeacons signal in the region of interest (ROI) in major organs (liver, kidney, spleen, heart, lung and tumor) at 5 days after nanobeacons injection. www.nature.com/scientificreports/ Figure 2. (A) Epi-fluorescence analysis of nude mice bearing orthotopic human gastric cancer cells, 0.5, 4, 24, 72 and 120 hours after tail-vein injection of nonsense (50 nM) and anti-Kras (5 and 50 nM) nanobeacons. Arrows represent gastric tumors. (B) Epi-fluorescence images of whole body organs to evaluate selective signal from the gold nanobeacons accumulated in tumors. (C) Quantification of anti-Kras nanobeacons signal in the region of interest (ROI) in major organs (liver, kidney, spleen, heart, lung and tumor) at 5 days after nanobeacons injection. can be highly captured by cells because of their repulsive interactions with the negatively charged cell surface; and are more likely to use caveolae-dependent endocytosis. y p y Representative images of whole body organs and resected tumors in mice treated with 5 and 50 nM of anti-Kras nanobeacons is depicted in Fig. 2B. Other organs showed residual accumulation, consistent with previous work reported for AuNPs28. These results showed that anti-Kras nanobeacons accumulate also in lungs. As evaluated by the quantification of nanobeacons signal the anti-Kras nanobeacons at 5 and 50 nM accumulate preferentially at the gastric tumor site and lungs (Fig. 2C). Quantification analy- sis of tumor ROI also revealed the specificity of the anti-Kras nanobeacons to the gastric tumors, when compared to nonsense nanobeacons (Supplementary Figure S7). Moreover, monitoring tumor ROI as Representative images of whole body organs and resected tumors in mice treated with 5 and 50 nM of anti-Kras nanobeacons is depicted in Fig. 2B. Other organs showed residual accumulation, consistent with previous work reported for AuNPs28. These results showed that anti-Kras nanobeacons accumulate also in lungs. www.nature.com/scientificreports/ As evaluated by the quantification of nanobeacons signal the anti-Kras nanobeacons at 5 and 50 nM accumulate preferentially at the gastric tumor site and lungs (Fig. 2C). Quantification analy- sis of tumor ROI also revealed the specificity of the anti-Kras nanobeacons to the gastric tumors, when compared to nonsense nanobeacons (Supplementary Figure S7). Moreover, monitoring tumor ROI as Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 5 ntificreports/ Figure 3. ATR-FTIR imaging of the tumor tissue for mice treated with naked nanoparticles (A), NB@ nonsense (B) and NB@Anti-Kras (D) on tumor region. (C) The spectra extracted from the tumor region with nonsense nanobeacons weaker bands at 1712, 1662, 1493 cm−1 and new bands at 1750 and 1100 cm−1. (E) The spectra extracted from the tumor region of mice treated with anti-Kras nanobeacons show stronger bands at 1712, 1662, 1493 and 1082 cm−1. www.nature.com/scientificreports/ Figure 3. ATR-FTIR imaging of the tumor tissue for mice treated with naked nanoparticles (A), NB@ nonsense (B) and NB@Anti-Kras (D) on tumor region. (C) The spectra extracted from the tumor region with nonsense nanobeacons weaker bands at 1712, 1662, 1493 cm−1 and new bands at 1750 and 1100 cm−1. (E) The spectra extracted from the tumor region of mice treated with anti-Kras nanobeacons show stronger bands at 1712, 1662, 1493 and 1082 cm−1. a function of time after treatment revealed a significant tumor size reduction using 50 nM of anti-Kras nanobeacons, when compared with 5 nM. 37h It is well known that metastasis occur frequently from gastric tumors to the lungs . Therefore, in order to track gold nanobeacons in tumor and lung tissues plasmonic-tunable Raman/FITR imaging spectroscopy was used (Fig. 3). Figure 3A shows the FTIR image of gold nanoparticles only. The cancer tissue was under processed with standard cryosection steps using regular histopathology methodologies. The sections were spread on 2 mm CaF2 substrate and remained embedded in paraffin, without any chemical treatment such as dewaxing. The reason for this is to prevent any further chemical modification Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 6 www.nature.com/scientificreports/ of the sample due to reagents and also to create some degree of refractive index matching, thus reducing the spectral distortion due to Mie scattering38. p g At the spectral region with frequencies 750–4000 cm−1, absorption was more intense for the anti-Kras nanobeacons treated mice. www.nature.com/scientificreports/ Other organs showed residual accumulation, consistent with previous work reported for AuNPs44.h The higher gold content in the tumor tissue by the anti-Kras nanobeacons may be due to a delay in nanoparticles recycling by the cells, once the anti-Kras nanobeacons may stay bound to the target mRNA. Once the nonsense nanobeacons do not present any function within the cell they excreted by the cell much faster than the anti-Kras nanobeacons. This fact is corroborated by the data on the nano- beacons’ in vitro uptake kinetics measured by ICP-MS (see Supplementary Figure S8).t To validate safety assessment, 5 days after injection with nanobeacons organs were harvested from mice and H&E stained for routine pathological analysis (Supplementary Figure S9). H&E staining showed that, in vivo administration of both nonsense and anti-Kras nanobeacons did not cause any significant damage in several organs (i.e. heart, liver, spleen, and kidney). g g p y However, histological analysis in tumor tissue showed evidence of extensive reduced vascularization for anti-Kras nanobeacons loaded when compared to nonsense nanobeacons (Fig. 5A and Supplementary Figure S9), in accordance with tumor size reduction (see Fig. 2B). Moreover, H&E staining of lung tissue show that nonsense nanobeacon exposed groups exhibited inflammatory accumulation at the broncho- alveolar junction and thickened local alveolar walls (Fig. 5A and Supplementary Figure S9). Significant reduction in severe interstitial infiltration of inflammatory cells is noted in the anti-Kras nanobeacon treated group. Nonsense nanobeacon treated group showed a predominance of mononuclear cells, as well as perivascular and peribronchiolar edema, occurrence of hypercellularity and thickened local alveolar walls and septa and increased number of tumoral clones (arrows in Fig. 5A). These histological images confirmed an evident decrease (~90%) in the incidence and severity of tumor clones and metastasis in lung from mice treated with anti-Kras nanobeacons, after 5 days of exposure. gt y Previous studies have been shown that metastatic cells could be established in the lungs before their oncogenic transformation, forming intravascular colonies in the lungs with concomitant invasion of the organ45,46. In fact, the incidence of pulmonary metastatic lung cancer from gastric cancer has been reported as approximately 50%, based on autopsy results36,47.i In order to confirm the reduction in number of gastric cancer metastasis into lung tissue, immuno- colocalization staining’s were performed. Representative confocal microscopy images of lung tissue, con- firmed the persistence of gastric tumor cells presence in the lungs (Fig. www.nature.com/scientificreports/ The spectrum extracted from the tumor region shows absorption peaks at 1712, 1662, 1493 and 1082 cm−1, representing the DNA phosphate backbone stretching vibrations of anti-Kras nanobeacons (Fig. 3E), while the spectrum extracted from the tumor region with nonsense nanobeacons shows weaker bands at 1712, 1662, 1493 and 1082 cm−1. The infrared band at 1712 cm−1 exemplifies the H-bonded C2 = O2 stretching vibration of thymines (T) from the third strand in TA–T triplex, where the third strand is involved in a reverse Hoogsteen base pair scheme. The band at 1662 cm−1 is attributed primarily to thymine stretching vibrations of C4 = O4 bonds. The band at 1493 cm−1 appears due to the ring stretching vibrations of cytosine (C = C) respectively39–42. In FTIR spectra of nonsense nanobeacons treated mice a new peak at 1750 cm−1 appears and the peak at 1082 cm−1 (that corresponds to the vibration intensity of the phosphoric acid groups (PO2−) with a symmetric stretching at, which has been extensively reported for several other material types43) is shifted to 1100 cm−1. Therefore, for the PO2− symmetric stretching frequency, the band from the anti-Kras nanobeacons treated mice exhibit a peak at 1082 cm−1 in the original profile (Fig. 3E), and the peak from the nonsense nanobeacons treated mice appeared at 1100 cm−1 (Fig. 3C). Higher spectra peaks can be observed in anti-Kras nanobeacons when compared to nonsense nanobeacon, probably due to higher values of free energy in secondary structure formation (Supplementary Figure S2). y g Spatial variations in the absorbance of these bands indicate the ability of micro ATR-FTIR imaging to probe chemical differences in tumor area (Fig. 3B,D). In addition to biological tissue, macro ATR-FTIR imaging has also been applied to study biomaterial processes including the plasmonic-tunable Raman/ FITR imaging spectroscopy, which results show that anti-Kras nanobeacons accumulate extensively in the tumor tissue when compared to the nonsense nanobeacons. Nevertheless, both nanobeacons accumulate similarly in lung tissue (Fig. 4A,B). In order to validate these data, inductively coupled plasma mass spectrometry (ICP-MS) showed that after 5 days of treat- ment, only anti-Kras nanobeacons accumulate extensively in tumor tissue; whereas both anti-Kras and nonsense nanobeacons accumulate in lung tissue (with a 5-fold increase when compared to other organs, such as heart, liver, spleen, kidney – Fig. 4C). Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 www.nature.com/scientificreports/ 5B) only in the group treated with nonsense nanobeacons. Alexa-Fluor® 594-labeled anti-human vimentin and FITC-labeled anti-human HLA antibodies specific for human antigens (that do not cross-react with either mouse fibroblasts or tumor cells) were used to visualize human gastric cancer metastasis in lung from mice treated with nanobeacons, as described elsewhere48. Vimentin is a type III intermediate filament (IF) protein that is expressed in mesenchymal cells, and is often used as a marker of mesenchymal-derived cells or cells undergoing an epithelial-to-mesenchymal transition (EMT) during both normal development and met- astatic progression49. On the other hand, the human leucocyte antigen (HLA) system is the major his- tocompatibility complex in humans, playing an essential role in the immune recognition of tumor cells that escape the immune system, and lead to lung metastasis for example50. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 7 www.nature.com/scientificreports/ Figure 4. (A) ATR-FTIR image of nonsense and anti-Kras nanobeacons showing the integrated absorbance at 1712 cm−1 on tumor and lung area. (B) Relative ATR-FTIR intensity at 1712 cm−1 for nonsense and anti- Kras nanobeacons accumulated in tumor and lung tissues. (C) Quantification of gold accumulated in tissues by inductively coupled plasma mass spectrometry (ICP-MS). Biodistribution data of nonsense and anti-Kras nanobeacons in major organs (liver, spleen, lung, kidney, heart and tumor) at 5 days after nanobeacons injection. Note the difference in tumor accumulation between the nonsense and anti-Kras nanobeacons and in comparison with the other major organs. Both nonsense and anti-Kras nanobeacons accumulate significantly in lungs. The SD (error bars) were calculated based on six animals (n = 6) per each study group. Figure 4. (A) ATR-FTIR image of nonsense and anti-Kras nanobeacons showing the integrated absorbance at 1712 cm−1 on tumor and lung area. (B) Relative ATR-FTIR intensity at 1712 cm−1 for nonsense and anti- Kras nanobeacons accumulated in tumor and lung tissues. (C) Quantification of gold accumulated in tissues by inductively coupled plasma mass spectrometry (ICP-MS). Biodistribution data of nonsense and anti-Kras nanobeacons in major organs (liver, spleen, lung, kidney, heart and tumor) at 5 days after nanobeacons injection. Note the difference in tumor accumulation between the nonsense and anti-Kras nanobeacons and in comparison with the other major organs. Both nonsense and anti-Kras nanobeacons accumulate significantly in lungs. The SD (error bars) were calculated based on six animals (n = 6) per each study group. www.nature.com/scientificreports/ Vimentin and HLA expression is much higher in lung tissue treated with nonsense nanobeacons, when compared to anti-Kras nanobeacons (Fig. 5B). These results corroborate the decrease in the num- ber of gastric cancer metastasis in lung tissue (~80% decrease) via the administration of anti-Kras nano- beacons (Fig. 5C). Vimentin and HLA expression is much higher in lung tissue treated with nonsense nanobeacons, when compared to anti-Kras nanobeacons (Fig. 5B). These results corroborate the decrease in the num- ber of gastric cancer metastasis in lung tissue (~80% decrease) via the administration of anti-Kras nano- beacons (Fig. 5C). g Next, we hypothesized that anti-Kras nanobeacons would enhance the likelihood of observing an increase in mice survival, as their administration results in significant tumor size reduction and in the decrease of number of metastatic lung cells. Accordingly, nude mice bearing human gastric xenograft tumors were treated with both nonsense and anti-Kras nanobeacons (50 nM). Animals treated with anti-Kras nanobeacons survived significantly longer (*P < 0.005 – Fig. 5D) compared to nonsense nanobeacon group. The results indicated that the observed survival extension of more than 85% could be attributed to both the inhibition of gastric tumor cells in the primary tumor and by the extensive reduction in the occurrence of metastasis in lung. The improved outcome in tumor-bearing mice receiv- ing anti-Kras nanobeacon treatment strongly supports the extraordinary potential of these nanoparticles as adjuvant agents to anticancer therapies. Effectively targeting metastases has proven to be extremely challenging due to the difficulty in overcoming biological barriers surrounding the cancer cells and the lack of tissue specificity. These bioresponsive antisense DNA gold nanobeacons prove to be a successful platform to target both cancer cells and metastasis. Our developed platform can be easily adjusted to hybridize with any specific target and provide rapid and cheap diagnosis and treatment for neoplastic diseases.fi Taken together, treatment with antisense DNA anti-Kras nanobeacons efficiently and selectively reduced the primary gastric tumor size by 60% and impaired the establishment of lung metastatic cells in more than 80% of the cases, thus increasing mice survival in more than 85%. Our data indicate that gas- tric tumor-associated cells shed from the primary tumor together with accompanying cancer cells survive to a secondary site and proliferate within the metastatic nodules in the lung in the absence of anti-Kras nanobeacons treatment. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 www.nature.com/scientificreports/ nanobeacons have a positive effect on metastatic cell colonization in the lungs in a syngeneic and spon- taneous mice model. In summary, the nanobeacons platform is of high therapeutic potential and serves as a theranostics and neoadjuvant therapy of orthotopic gastric tumors but also as anti-metastasis treat- ment, where nanobeacons show a strong involvement in both local and distant lesions. This platform can be further designed to selectively and specifically silence any target gene to overcome a broad range of cancerous diseases. www.nature.com/scientificreports/ Following selective Kras inhibition via antisense DNA nanoprobes, these unique 8 Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 5. (A) Histological analysis of gastric tumor and lung tissue indicating a reduced vascularization in the primary tumor and a decrease in tumoral clones in lung tissue treated with anti-Kras nanobeacons only. Nonsense nanobeacons group present inflammatory cells accumulation at bronchoalveolar junction and thickened local alveolar walls and high prevalence of tumoral clones (arrows). (B) Representative confocal microscopy images of lung tissue immunostaining, confirming the persistence of gastric cancer metastasis in the lungs. Blue: DAPI nuclear stain; red: Alexa-Fluor® 594-labeled anti-human vimentin; green: FITC-labeled anti-human HLA. Scale bars, 20 μ m. (C) anti-Kras nanobeacons treatment significantly reduced the number of spontaneous lung metastasis in gastric cancer model (P < 0.005, n = 6 mice). (D) Kaplan-Meier survival curves for mice treated with nonsense (black) and anti-Kras (red) nanobeacons using Log-Rank test. Data points represent group mean ± SD (n = 6, P < 0.005). Figure 5. (A) Histological analysis of gastric tumor and lung tissue indicating a reduced vascularization in the primary tumor and a decrease in tumoral clones in lung tissue treated with anti-Kras nanobeacons only. Nonsense nanobeacons group present inflammatory cells accumulation at bronchoalveolar junction and thickened local alveolar walls and high prevalence of tumoral clones (arrows). (B) Representative confocal microscopy images of lung tissue immunostaining, confirming the persistence of gastric cancer metastasis in the lungs. Blue: DAPI nuclear stain; red: Alexa-Fluor® 594-labeled anti-human vimentin; green: FITC-labeled anti-human HLA. Scale bars, 20 μ m. (C) anti-Kras nanobeacons treatment significantly reduced the number of spontaneous lung metastasis in gastric cancer model (P < 0.005, n = 6 mice). (D) Kaplan-Meier survival curves for mice treated with nonsense (black) and anti-Kras (red) nanobeacons using Log-Rank test. Data points represent group mean ± SD (n = 6, P < 0.005). nanobeacons have a positive effect on metastatic cell colonization in the lungs in a syngeneic and spon- taneous mice model. In summary, the nanobeacons platform is of high therapeutic potential and serves as a theranostics and neoadjuvant therapy of orthotopic gastric tumors but also as anti-metastasis treat- ment, where nanobeacons show a strong involvement in both local and distant lesions. This platform can be further designed to selectively and specifically silence any target gene to overcome a broad range of cancerous diseases. Methods Synthesis Synthesis of citrate gold nanoparticles. Gold nanoparticles, with an average diameter of 13.2 ± 2.1 nm, were synthesized by the citrate reduction method described by Lee and Meisel51. Briefly, 225 mL of 1 mM hydrogen tetrachloroaureate (III) hydrate (Sigma) (88.61 mg) was solved in 500 ml of distilled water, heated, and stirred under reflux. When the solution boils, 25 mL of 38.8 mM sodium citrate dihydrate (Sigma) (285 mg) was added resulting in a red solution. The solution is kept under ebullition with vigorous stirring and protected from light for 30 minutes. After this, the solution is cooled Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 9 www.nature.com/scientificreports/ down and kept protected from light. Citrate gold nanoparticles were characterized by Transmission Electron Microscopy (TEM) and UV-Vis molecular absorption spectra. down and kept protected from light. Citrate gold nanoparticles were characterized by Transmission Electron Microscopy (TEM) and UV-Vis molecular absorption spectra. Synthesis of PEGylated-Gold nanoparticles. Functionalization of PEGylated gold nanoparticles was carried out using commercial hetero-functional poly(ethylene glycol) (PEG) functionalized with a 30% saturated surface of α -Mercapto-ω -carboxy PEG solution (HS-C2H4-CONH-PEG-O-C3H6-COOH, MW. 3500 Da, Sigma) as described elsewhere52,53. The 30% of saturated PEG layer allows the incorpo- ration of additional thiolated components, such as the thiolated DNA-hairpin-Cy3. Briefly, 10 nM of the citrate-gold nanoparticles were mixed with 0.006 mg/mL of PEG solution in an aqueous solution of SDS (0.028%). After this, the mixture was incubated for 16 hours at room temperature. Excess PEG was removed by centrifugation (15,000 × rpm, 30 min, 4 °C), and quantified by a modification of the Ellman’s Assay (see Supplementary Information S2 for complete protocol and quantification methods). Synthesis of Gold Nanobeacons. Two different sequences of Gold Nanobeacons were prepared: a nanobeacon anti-Kras (detects and inhibits Kras mRNA) and a nanobeacon nonsense (designed to not hybridizes with any target within the genome) (Supplementary Table S1 for thiol-DNA-hairpin Cy3 sequences). Briefly, the thiolated oligonucleotides (Sigma) – thiol-DNA-hairpin Cy3 were suspended in 1 mL of 0.1 M dithiothreitol (DTT, Sigma), extracted three times with ethyl acetate and further puri- fied through a desalting NAP-5 column (GE Healthcare) using 10 mM phosphate buffer (pH 8) as elu- ent. Following oligonucleotide quantification via UV/Vis spectroscopy, each oligomer was added to the AuNP-PEG solution in a 50:1 ratio. Methods Synthesis AGE I solution (2% (w/v) SDS, 10 mM phosphate buffer (pH 8)) was added to the mixture to achieve a final concentration of 10 mM phosphate buffer (pH 8), 0.01% (w/v) SDS. The solution was sonicated for 10 seconds using an ultrasound bath and incubated at room temper- ature for 20 minutes. Afterwards, the ionic strength of the solution was increased sequentially in 50 mM NaCl increments by adding the required volume of AGE II solution (1.5 M NaCl, 0.01% (w/v) SDS, 10 mM phosphate buffer (pH 8)) up to a final concentration of 10 mM phosphate buffer (pH 8), 0.3 M NaCl, 0.01% (w/v) SDS. After each increment, the solution was sonicated for 10 seconds and incubated at room temperature for 20 minutes before the next increment. Following the last addition, the solution was left to rest for additional 16 hours at room temperature. Then, the functionalized gold nanobeacons were centrifuged for 20 minutes at 15,000 × rpm, the oily precipitate washed three times with MilliQ water, and redispersed in MilliQ water. The resulting gold nanobeacons were stored in the dark at 4 °C until further use. Characterization of the gold nanobeacons was performed by Dynamic Light Scattering – DLS (Wyatt Dyna Pro Plate Reader), UV/Vis Spectroscopy and TEM (see Supplementary Table S2). Specificity assays for Gold Nanobeacons. For the detection of specific targets, 2.5 nM of nanobea- con anti-Kras and nanobeacon nonsense in 10 mM of phosphate buffer pH 7 was added to 5 nM of com- plementary Kras target (Sigma). All measurements were performed in a microplate reader (Varioskan Flash Multimode Reader, Thermo Scientific) programmed to incubate the reactions for 120 min at 37 °C while recording the fluorescence intensity every 2 minutes at an excitation wavelength of 550 nm for Cy3-labeled gold nanobeacon. In vitro Gold Nanobeacon’s delivery. Human gastric cancer MGC-803 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) supplemented with 4 mM glutamine, 10% heat inactivated fetal bovine serum (Invitrogen), 100 U/ml penicillin and 100 μ g/ml streptomycin (Invitrogen) and maintained at 37 °C in 5% CO2. Cells were seeded at a density of 1 × 105 cells/well in 24-well plates and grown for 24 hours prior to incubation of nanobeacons. On the day of incubation, the cells were approximately 50% confluent. For flow cytometry, data were acquired on FACS LSR Fortessa HTS-1 (BD Biosciences) flow cytometer. Quantitative PCR. Methods Synthesis Total RNA from Human gastric cancer cell line MGC-803 was extracted using RNeasy Plus Mini Kit (Qiagen) according to the manufacture’s protocol. cDNA was produced using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems) using 500 ng of total RNA. qRT-PCR was performed with Taqman probes FAM-MGB for Kras and GAPDH (Applied Biosystems). GAPDH was used as a reference gene. The reactions were processed using Light Cycler 480 II Real-time PCR machine (Roche) using TaqMan® Gene Expression Master Mix (Applied Biosystems) under the following cycling steps: 2 min at 50 °C for UNG activation; 10 min at 95 °C; 40 cycles at 95 °C for 15 s; 60 °C for 60 s. At least three independent repeats for each experiment were carried out. Gene expression was determined as a difference in fold after normalizing to the housekeeping gene GAPDH. Construction of orthotopic gastric cancer mice model. All animal experiments were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University (NO.SYXK2007- 0025). The animal experimentation methods were carried out in accordance with the approved and relevant guidelines and regulations. Nude mice (nu/nu, female, 18 g, 4 weeks old) were purchased from the Shanghai SLAC Laboratory animal Co. Ltd and housed in SPF grade animal center. Subcutaneous gastric cancer (GC) tissues at the exponential growth phase loaded by nude mice were resected asepti- cally. Necrotic tissues were cut away, and the remaining tumor tissues were scissor-minced into pieces Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 10 www.nature.com/scientificreports/ around 1–2 mm in diameter in 4 °C Hanks’ balanced salt solution, and each piece was adjusted to be 30 mg with scissors. Before the surgery, all mice were fasting for at least 8 hours. Mice were anesthetized with 5% trichloraldehyde hydrate, with 4 μ L/g weight and an incision was made through the left upper abdominal pararectal line and peritoneum. The stomach wall was carefully exposed, and a part of the serosal membrane, about 2 mm in diameter, in the middle of the greater curvature of the glandular stomach was mechanically scratched with ophthalmic knives. A tumor piece of 30 mg was then fixed on the injured site of the serosal surface with OB glue (Guang Zhou Baiyun Ltd, China). The stomach was then returned to the peritoneal cavity, and the abdominal wall and skin were closed with 9-0 ophthalmic sutures. Methods Synthesis The animals were kept on warm electric blanket at 37 °C until analepsia and kept separately in an SPF environment. FTIR Instrumentation. ATR spectra were recorded with the Perkin Elmer Spotlight 400 N Universal Attenuated Total Reflectance (UATR) accessory of the spectrometer, which employs a 9-bounce dia- mond top-plate for this analysis. Sample penetration is both wavenumber and sample dependent, but is typically of at the order of 1 mm. In ATR mode, spectral data were the result of 4 scans, with a spectral resolution of 8 cm−1. Tissue samples were seed on CaF2 crystal and air dry for 10 min prior to recording. A background spectrum was also recorded and automatically subtracted by the software. Analysis of tumor growth and mice survival. Non-invasive longitudinal monitoring of tumor progression was followed by scanning mice with the Bruker In-Vivo F PRO imaging system from mice bearing gastric tumors (n = 6 animals per treated group). Whole-animal imaging was performed at the indicated time points – 0.5, 4, 24, 72 and 120 hours after nanobeacons injection. Kaplan-Meier survival curves were designed by evaluating mice survival follow up. In order to compare the survival distribu- tions of mice treated with anti-Kras and nonsense nanobeacons log-rank tests were performed. Histology and immunolocalisation. After mice have been sacrificed and perfused with sterile PBS, organs were snap-frozen in optimum cutting temperature compound (Tissue-Tek® ) and cut on a cryostat microtome and air-dried overnight. Frozen sections were thawed and stained for hematoxylin and eosin using standard procedures. Stained tissue sections were then imaged by light microscopy. For immu- nolocalisation, the tumor and lung tissues were embedded and sliced into a 3 μ m section. After fixation with 4% paraformaldehyde in PBS, lung tissue sections were treated with blocking solution containing 4% rabbit serum in PBS. For metastasis detection, the lung tissue slide was incubated with Alexa Fluor® 594-labeled anti-human vimentin antibody (1:250 Abcam) and FITC-labeled anti-human HLA antibody (1:100, BioLegend) in PBS containing 1% BSA for 60 min, and was washed three times in PBS. Nuclei were DAPI stained before slide mounting with Fluoromount™ Aqueous Mounting Medium (Sigma). Stained tissue sections were then imaged by fluorescence microscopy. Biodistribution studies. quantification of gold nanobeacons accumulation in the tumor and other organs. Methods Synthesis Biodistribution of the functionalized gold nanobeacons in tissues associated with clearance (liver, spleen and kidney) as well as the gastric tumor, lung and heart was measured by inductively coupled plasma mass spectrometry (ICP-MS). Briefly, 5 days after gold nanobeacons administration, the mice were sacrificed to harvest the major organs, which were rinsed with ethanol three times and then air-dried into clean vials for acid digestion (aqua-regia 3HCl:1HNO3). After 1 day of strong acid diges- tion, the samples were analyzed by ICP-MS. Statistics. Differences between groups were examined using Student’s paired t test through SPSS sta- tistical package (version 17, SPSS Inc.). All error bars used in this report are mean ± SD of at 3 independ- ent experiments. Statistically significant P values were indicated in figures and/or legends as P < 0.005; P < 0.01; P < 0.05. All in vivo experiments used 5 mice per treatment group unless noted otherwise. 3. 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Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 12 www.nature.com/scientificreports/ Acknowledgementsh g This work was funded in part by Science Foundation Ireland under Grant Number 11/PI/08, the National Key Basic Research Program (973 Project) (No. 2011CB933101 and 2015CB931802), National Natural Scientific Fund (No. 81225010), 863 project of China (2012AA022703 and 2014AA020700), Shanghai Science and Technology Fund (No. 13NM1401500 ). JC acknowledges Marie Curie International Outgoing Fellowship (FP7-PEOPLE-2013-IOF, Project no. 626386) and FT for Marie Curie grant Agreement (PIEF-GA-2012-332-332462). The authors also thank to the Peterson Nanotechnology Materials Core Facility and the Swanson Biotechnology Center at the Koch Institute (MIT) and the FACS Core Center with Cancer Center Support Grant P30-CA14051 from the NCI. Author Contributions J. Conde, F.T. and C.B. conceived the project and designed the experiments. J. Conde, F.T. and C.B. performed the experiments, collected and analysed the data. J. Conde, F.T., D.C., N.A. and J. Curtin co- wrote the paper. All authors discussed the results and reviewed the manuscript. Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 Additional Information Supplementary information accompanies this paper at http://www.nature.com/srepi Competing financial interests: The authors declare no competing financial interests. How to cite this article: Bao, C. et al. Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis. Sci. Rep. 5, 12297; doi: 10.1038/srep12297 (2015). How to cite this article: Bao, C. et al. Bioresponsive antisense DNA gold nanobeacons as a hybrid in vivo theranostics platform for the inhibition of cancer cells and metastasis. Sci. Rep. 5, 12297; doi: 10.1038/srep12297 (2015). 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 Com- mons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ Scientific Reports \| 5:12297 \| DOI: 10.1038/srep12297 13	7,523
https://openalex.org/W2766139506	OpenAlex	Open Science	CC-By	2,017	PREDICTING THE RISK OF PROTHROMBOTIC CHANGES IN ADOLESCENTS WITH ESSENTIAL HYPERTENSION	Svetlana Bolshakova	Russian	Spoken	4,128	12,486	ACTA BIOMEDICA SCIENTIFICA, 2017, Vol. 2, N 4 УДК 616-053.5+616.151.511+616.1 ACTA BIOMEDICA SCIENTIFICA, 2017, Vol. 2, N 4 УДК 616-053.5+616.151.511+616.1 Большакова С.Е., Гомелля М.В., Баирова Т.А., Бердина О.Н., Бугун О.В., Михалевич И.М., Рычкова Л.В. Большакова С.Е., Гомелля М.В., Баирова Т.А., Бердина О.Н., Бугун О.В., Михале Рычкова Л.В. ПРОГНОЗИРОВАНИЕ РИСКА ВОЗНИКНОВЕНИЯ ПРОТРОМБОТИЧЕСКИХ ИЗМЕНЕНИЙ У ПОДРОСТКОВ С ЭССЕНЦИАЛЬНОЙ АРТЕРИАЛЬНОЙ ГИПЕРТЕНЗИЕЙ ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (664003, г. Иркутск, ул. Тимирязева, 16, Россия) Сердечно-сосудистые заболевания (ССЗ) остаются важнейшей причиной смертности во всем мире. Наиболее распространённым ССЗ является эссенциальная артериальная гипертензия (ЭАГ), рост заболеваемости которой отмечается среди детей, особенно среди подростков. Исследования по изучению нарушений в системе гемостаза у больных с ЭАГ свидетельствуют о возможности формирования протромботических изменений (ПТИ). Однако проблема ранней диагностики ПТИ у подростков с ЭАГ является на сегодняшний день актуальной и требующей внимания педиатров. Цель исследования – разработать прогностические критерии риска возникновения ПТИ у подростков с ЭАГ для своевременного проведения профилактических мероприятий и предупреждения развития тромботических осложнений. Обследовано 37 подростков с ЭАГ и ПТИ и 60 – с ЭАГ, в возрасте 14–17 лет, проживающих на территории Восточной Сибири, у которых изучали особенности генеалогического анамнеза, показателей суточного мониторирования уровня артериального давления (СМАД), коагуляционные нарушения системы гемостаза и структуру полиморфизмов генов предрасположенности к тромбозам по данным клинико-анамнестического, функционального и лабораторного исследований. Контрольную выборку составили здоровые подростки, сопоставимые по полу и возрасту с пациентами клинических групп. В результате проведённого исследования создана модель прогнозирования риска развития ПТИ, позволяющая с точностью 86,18 % выявлять тромбогенный риск в подростковом возрасте у лиц с ЭАГ по определённым признакам (полиморфизм С677Т гена MTHFR, полиморфизм А66G гена MTRR, ДАДд, САДн, ИВ гипертензии САДд, отягощённый тромботический генеалогический анамнез, раннее наступление тромбозов, ИО тромботического анамнеза) и своевременно начать проведение профилактических мероприятий с целью снижения частоты тромботических осложнений ЭАГ и смертности больных. Ключевые слова: подростки, гены, эссенциальная артериальная гипертензия, протромботические изменения, тромбоз Bolshakova S.E., Gomellya M.V., Bairova T.A., Berdina O.N., Bugun O.V., Mikhalevich I.M., Rychkova L.V. Scientific Centre for Family Health and Human Reproduction Problems (664003, Irkutsk, ul. Timiryazeva, 16, Russian Federation) Today, the problem of early diagnosis of hematological changes predisposing to the development of thrombotic complications in patients with essential hypertension (EH) is an urgent problem that requires close attention not only of physicians, but also of pediatricians. The aim of the study was the development of prognostic criteria for risk of prothrombotic changes (PС) in adolescents with EH, timely preventive measures and prevention of thrombotic complications. Sixty аdolescents with EH without PС and 37 adolescents with EH and PС were examined. We used the following methods: clinical anamnestic (including genealogy), functional and ultrasound, laboratory, mathematical and statistical. To create a mathematical model of forecasting, discriminant analysis was used, with the help of which from the 59 proposed predictors the algorithm selected 8 most informative features: the C777T polymorphism of the 5,10-methylenetetrahydrofolate reductase gene, the A66G gene of the methionine synthase reductase gene, the daily diastolic blood pressure level, the level of the nocturnal systolic arterial pressure-time index of hypertension, systolic blood pressure during the day, weighed down by thrombotic genealogically history, early onset of thrombosis, burdened thrombotic genealogical history. Our method for predicting the risk of developing PС allows to place adolescents with EН having an increased risk of developing these coagulation shifts in a separate group, to identify thrombogenic risk in adolescence and, if necessary, to initiate preventive measures in time to reduce the incidence of thrombotic compli cations of EН and mortality of patients. Key words: adolescents, genes, essential hypertension, prothrombotic changes, thrombosis ВВЕДЕНИЕ инсульта, которые, по прогнозам, останутся един ственными основными причинами смерти [1]. Сердечно-сосудистые заболевания (ССЗ) остаются важнейшей причиной смертности во всем мире [8]. В 2012 г. от ССЗ умерли 17,5 млн человек, что состави ло 31 % всех случаев смерти в мире. Из этого числа 7,4 млн человек умерли от ишемической болезни сердца и 6,7 млн человек – в результате инсульта. По оценкам ВОЗ, к 2030 г. от ССЗ умрёт около 23,3 млн человек – главным образом, от болезней сердца и Наиболее распространённым ССЗ является эс сенциальная артериальная гипертензия (ЭАГ), рост заболеваемости которой отмечается среди детей, особенно среди подростков [3]. Осложнения данной патологии, в том числе со стороны системы гемостаза, все чаще регистрируются не только у людей старшего возраста, но и у молодых и даже у подростков [4, 9, 12]. Pediatrics 58 ACTA BIOMEDICA SCIENTIFICA, 2017, Том 2, № 4 Существует большое количество работ, посвящённых изучению различных изменений и маркеров, ассоци ированных с развитием ЭАГ и её осложнений [2, 5, 7, 10, 11]. Тем не менее, на сегодняшний день чётко не определены факторы, играющие роль в возникно вении протромботических изменений (ПТИ) у под ростков с ЭАГ, предшествующих тромбозу. Кроме того, в литературе не встречаются работы, посвящённые прогнозированию риска развития ПТИ у подростков с ЭАГ. Поэтому данная проблема на сегодняшний день является достаточно актуальной и требующей дальнейшего изучения. кривой распределения АД для соответствующего возраста, пола и роста. В ходе проведённого исследования применялись следующие методы: клинико-анамнестический, в том числе генеалогический; функциональные и ультразвуковые; лабораторные; математико-ста тистические. Особое внимание при сборе анамнеза жизни уделялось наличию тромботического анамне за в семье с уточнением линии родства. Для количе ственной оценки отягощённости генеалогического тромботического анамнеза рассчитывали индекс отягощённости (ИО), определяемый отношением общего количества эпизодов тромбоза у кровных родственников пробанда, о которых есть сведения (А), к общему числу всех родственников (Б), исклю чая пробанда (ИО = А / Б). При значении ИО = 0–0,2 степень отягощённости анамнеза считали низкой, при ИО = 0,3–0,5 – умеренной, при ИО = 0,6–0,8 – вы раженной, при ИО = 0,9 и выше – высокой. Оценка функционального состояния сердечно-сосудистой системы включала электрокардиографию в 12 стан дартных отведениях (аппарат FCP-4101U Fukuda Denshi, Япония) доплерэхокардиографию (аппарат Aloka SSD 1400), суточное мониторирование уровня АД (СМАД) в течение 24 часов (аппарат Oscar 2 для системы OXFORD Medilog Prima). ВВЕДЕНИЕ Лабораторные ме тоды исследования: оценка показателей коагуляци онного гемостаза на автоматическом коагулометре STA-R Evolution (Roche Diagnostics, Швейцария) с на борами реагентов фирмы Roche (Швейцария) (акти вированное парциальное тромбопластиновое время, протромбиновое время в % по Квику, тромбиновое время, концентрация фибриногена, уровень рас творимых фибрин-мономерных комплексов); опре деление полиморфизмов генов предрасположен ности к тромбозам методом полимеразной цепной реакции с последующим изучением полиморфизма длин рестрикционных фрагментов с использова нием комплектов реагентов «SNP-экспресс» фирмы «Литех» (Россия) по протоколу производителя. Статистическая обработка данных проводилась на персональном компьютере с помощью электронных таблиц Excel и пакета прикладных программ Biostat и STATISTICA 6.0 (StatSoft Inc., США). Разработку ма тематической модели индивидуального прогнози рования риска проводили на кафедре информатики ИГМАПО (заведующий кафедрой к.г.-м.н., доцент Михалевич И.М.) с помощью дискриминантного анализа. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ На базе клиники ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (г. Иркутск) в период с 2009 по 2012 гг. проведено обследование 97 подростков (14–17 лет): 60 подростков с ЭАГ, не имеющие ПТИ (71,7 % мальчиков и 28,3 % девочек), и 37 подростков с ЭАГ, имеющие ПТИ (67,6 % мальчи ков и 32,4 % девочек). Группы исследования значимо не различались по возрасту и полу. Размер выборки предварительно не рассчитывался. Исследование было выполнено в соответствии со стандартами надлежащей клинической практики (Good Clinical Practice) и принципами Хельсинской декларации. Получение письменного информированного согласия на участие в проводимом исследовании являлось обязательной процедурой до включения подростка в одну из групп наблюдения. Протокол исследования был одобрен Комитетом по биомедицинской этике при ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека». Диагноз ЭАГ установлен на основании клини ко-инструментальных критериев в соответствии с современной классификацией, разработанной экс пертной группой Всероссийского научного общества кардиологов (ВНОК, 2003) и Ассоциацией детских кардиологов России (2003). Критериями исключения из исследования яв лялись: наличие симптоматической (вторичной) формы артериальной гипертензии; наличие «ги пертонии белого халата»; наличие вегетососудистой дистонии по гипертензивному типу; неславянская этническая группа и подростки от смешанных бра ков; приём лекарственных препаратов. Группу кон троля составляли подростки, у которых на момент осмотра не было отмечено острых и хронических заболеваний, которые в двух предыдущих поко лениях не имели родственников с артериальной гипертензией и неславянской этнической группы, не предъявляли жалоб на повышение уровня арте риального давления (АД), и показатели АД у которых при многократном измерении и по данным суточного мониторирования не превышали 89-го перцентиля ЦЕЛЬ РАБОТЫ Разработать прогностические критерии риска возникновения ПТИ у подростков с ЭАГ для своев ременного проведения профилактических меропри ятий и предупреждения развития тромботических осложнений. РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ Все подростки с ЭАГ, включённые в исследование, имели определённый набор признаков, полученных в результате проведённого обследования, которые, по нашим данным и данным литературы, могут высту пать в качестве факторов риска (ФР) развития ПТИ. Исходно нами был рассмотрен набор из 59 признаков, объединённых в несколько групп: клинико-анамне стические данные (тромботический генеалогический анамнез, акушерский анамнез матерей подростков, данные о течении антенатального, интранатального и неонатального периодов, сопутствующие заболева Педиатрия Педиатрия 59 ACTA BIOMEDICA SCIENTIFICA, 2017, Vol. 2, N 4 – 9,98 %; раннее наступление тромбозов – 13,68 %; ИО тромботического анамнеза – 11,68 %. – 9,98 %; раннее наступление тромбозов – 13,68 %; ИО тромботического анамнеза – 11,68 %. – 9,98 %; раннее наступление тромбозов – 13,68 %; ИО тромботического анамнеза – 11,68 %. ния), данные функционального обследования (пара метры СМАД), данные молекулярно-генетического обследования пациентов. Для определения принадлежности объекта (пациента) к определенной группе необходимо рас считать значения F по формулам: Для создания математической модели про гнозирования был применён многомерный метод статистического анализа – дискриминантный ана лиз. Это один из методов решения так называемой задачи классификации – задачи отнесения изучае мого объекта (пациента) к одной из групп на основе измерения у него некоторого числа признаков, а также определение «веса» каждой дискриминантной переменной для разделения объектов на классы. При использовании данного метода алгоритм выбирает такие признаки, значения которых наиболее близки у объектов исследования одной группы и наиболее различны у объектов исследования из разных групп. В итоге выявляются и включаются в модель наиболее информативные признаки [6]. F1 = –0,59 – 0,42 × х1 + 0,21 × х2 – 0,30 × х3 + 0,29 × х4 – – 0,42 × х5 + 0,28 × х6 – 0,34 × х7 – 0,42 × х8, F2 = –2,44 + 1,01 × х1 – 0,69 × х2 + 0,95 × х3 – 0,76 × х4 + + 0,75×х5 – 0,77 × х6 + 1,11 × х7 + 0,82 × х8, F1 = –0,59 – 0,42 × х1 + 0,21 × х2 – 0,30 × х3 + 0,29 × х4 – – 0,42 × х5 + 0,28 × х6 – 0,34 × х7 – 0,42 × х8, F2 = –2,44 + 1,01 × х1 – 0,69 × х2 + 0,95 × х3 – 0,76 × х4 + + 0,75×х5 – 0,77 × х6 + 1,11 × х7 + 0,82 × х8, 5 6 7 8 где х1 – полиморфизм С677Т гена MTHFR; х2 – полиморфизм А66G гена MTRR; х3 – ДАДд; х4 – САДн; х5 – ИВ гипертензии САДд; х6 – отягощённый тромботический генеалогический анамнез; х7 – раннее наступление тромбозов; х8 – ИО тромботического анамнеза. Значения х1–х8 стандартизированы. При этом при F1 > F2 делается благоприятный прогноз и вероятность развития ПТИ подростков с ЭАГ считается низкой, а при F2 > F1 делается неблагоприятный прогноз и вероятность развития ПТИ подростков с ЭАГ считается высокой. Педиатрия Исходные данные были следующие: n = 97, m = 59, где n – количество подростков, принявших участие в исследовании; m – количество признаков. При сравнении групп с помощью дискриминант ного анализа из 59 предложенных предикторов алго ритм отобрал 8 наиболее информативных признаков, для которых уровень значимости по F-критерию р < 0,100 (табл. 1). Математическая модель, полученная в ходе применения дискриминантного анализа, требует дальнейшей проверки на работоспособность (вали дизация модели). Процедура проверки заключается в применении уравнений к новым объектам исследо вания, не включённым ранее в исследование (экзаме нующая группа), с целью оценки ее эффективности и устойчивости результатов распознавания. Мера Махаланобиса, с помощью которой оценива ется сумма квадратов расстояния между значениями канонических величин у обследованных подростков, равна 5,19 (р < 0,0001), что характеризует статистиче ски значимое различие между группами подростков. Суммарный показатель правильной классификации в группе подростков с ЭАГ и ПТИ составил 76,00 %, в группе подростков с ЭАГ без ПТИ – 96,36 %, средний – 86,18 %, что является высоким значением диффе ренциации групп. Мера Махаланобиса, с помощью которой оценива ется сумма квадратов расстояния между значениями канонических величин у обследованных подростков, равна 5,19 (р < 0,0001), что характеризует статистиче ски значимое различие между группами подростков. В нашей работе экзаменующую группу соста вили 14 подростков. На данной группе применили описанные выше дискриминантные уравнения. Среди подростков из группы с ЭАГ и ПТИ в 6 (75,0 %) случаях развитие изменений в системе гемостаза было предсказано верно, у 2 (25,0 %) – неверно. У всех подростков из группы с ЭАГ без ПТИ прогноз оказался верным. В нашей работе экзаменующую группу соста вили 14 подростков. На данной группе применили описанные выше дискриминантные уравнения. Среди подростков из группы с ЭАГ и ПТИ в 6 (75,0 %) случаях развитие изменений в системе гемостаза было предсказано верно, у 2 (25,0 %) – неверно. У всех подростков из группы с ЭАГ без ПТИ прогноз оказался верным. Суммарный показатель правильной классификации в группе подростков с ЭАГ и ПТИ составил 76,00 %, в группе подростков с ЭАГ без ПТИ – 96,36 %, средний – 86,18 %, что является высоким значением диффе ренциации групп. Вклад каждого признака в различие был следую щим: полиморфизм С677Т гена MTHFR – 13,52 %; по лиморфизм А66G гена MTRR – 8,45 %; ДАДд – 11,80 %; САДн – 9,83 %; ИВ гипертензии САДд – 11,06 %; отяго щённый тромботический генеалогический анамнез ВЫВОДЫ 1. Предложенные математико-статистические построения с точностью 86,18 % дают возможность REFERENCES Gaeta M, Campanella F, Gentile L, Schifino GM, Ca passo L, Bandera F, Banfi G, Arpesella M, Ricci C. (2017). European cardiovascular mortality over the last three decades: evaluation of time trends, forecasts for 2016. 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Acetylsalicylic acid in the prevention and treatment of cardiovascular diseases: clinical recommendations for practicing doctors [Atsetil salitsilovaya kislota v profilaktike i lechenii serdechno-so sudistykh zabolevaniy: klinicheskie rekomendatsii dlya praktikuyushchikh vrachey]. Russkiy meditsinskiy zhurnal, (14), 708-714. Zhitnikova LM. (2012). Acetylsalicylic acid in the prevention and treatment of cardiovascular diseases: clinical recommendations for practicing doctors [Atsetil salitsilovaya kislota v profilaktike i lechenii serdechno-so sudistykh zabolevaniy: klinicheskie rekomendatsii dlya praktikuyushchikh vrachey]. Russkiy meditsinskiy zhurnal, (14), 708-714. спрогнозировать развитие ПТИ у подростков с ЭАГ по определённым признакам (полиморфизм С677Т гена MTHFR, полиморфизм А66G гена MTRR, ДАДд, САДн, ИВ гипертензии САДд, отягощенный тромботический генеалогический анамнез, раннее наступление тром бозов, ИО тромботического анамнеза). 2. Разработанный метод прогнозирования риска возникновения ПТИ позволяет выделить группу подростков с ЭАГ, имеющих повышенный тромбо генный риск, и своевременно начать проведение профилактических мероприятий с целью снижения частоты тромботических осложнений ЭАГ и смерт ности больных. 5. Колесникова Л.И., Долгих В.В., Поляков В.М., Рычкова Л.В. Проблемы психосоматической пато логии детского возраста. – Новосибирск: Наука, 2005. – 221 c. Kolesnikova LI, Dolgikh VV, Polyakov VM, Rychkova LV. (2005). Problems of psychosomatic pathology of child hood [Problemy psikhosomaticheskoy patologii detskogo vozrasta]. Novosibirsk, 221 p. Авторы данной статьи сообщают об отсутствии конфликта интересов. 6. Михалевич И.М., Алферова М.А., Рожкова Н.Ю. Основы прикладной статистики: учебное пособие. – Иркутск: РИО ИГИУВа, 2008. – 71 c. у р , ; ) Bolshakova Svetlana Evgenyevna – Candidate of Medical Sciences, Research Officer at the Laboratory of Pediatrics and Neuro physiology of Scientific Centre for Family Health and Human Reproduction Problems (664003, Irkutsk, ul. Timiryazeva, 16; e-mail: sebol@bk.ru) Большакова Светлана Евгеньевна – кандидат медицинских наук, научный сотрудник лаборатории педиатрии и нейрофизиологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (664003, г. Иркутск, ул. Тимирязева, 16; e-mail: sebol@bk.ru) Таблица 1 Таблица 1 Признаки, используемые для дискриминантного анализа n = 97 Wilks' Lambda: ,47271 approx. F (8,71) = 9,8996 p < 0,00001 Wilks' Частичные F-remove p-level Toler. 1-Toler. С677Т 0,557114 0,848505 12,67655 0,000666 0,843675 0,156325 A66G 0,510499 0,925984 5,67516 0,019888 0,883839 0,116161 ДАДд 0,522945 0,903945 7,54456 0,007622 0,670612 0,329388 САДн 0,508478 0,929666 5,37153 0,023355 0,633020 0,366980 ИВ САДд 0,513296 0,920939 6,09521 0,015963 0,690371 0,309629 Отягощенный тромботический анамнез 0,484779 0,975112 1,81216 0,182532 0,229941 0,770058 Раннее наступление тромбозов 0,529788 0,892271 8,57224 0,004584 0,533750 0,466250 ИО 0,489867 0,964985 2,57631 0,112914 0,256084 0,743916 Примечание. С677Т – полиморфизм С677Т гена MTHFR, полиморфизм A66G гена MTRR; ДАДд – диастолическое артериальное давление среднедневное; САДн – систолическое артериальное давление средненочное; ИВ САДд – индекс времени гипертензии систолического артериального давления днём. Признаки, используемые для дискриминантного анализа 60 ACTA BIOMEDICA SCIENTIFICA, 2017, Том 2, № 4 Сведения об авторах Information about authors Большакова Светлана Евгеньевна – кандидат медицинских наук, научный сотрудник лаборатории педиатрии и нейрофизиологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (664003, г. Иркутск, ул. Тимирязева, 16; e-mail: sebol@bk.ru) Большакова Светлана Евгеньевна – кандидат медицинских наук, научный сотрудник лаборатории педиатрии и нейрофизиологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (664003, г. Иркутск, ул. Тимирязева, 16; e-mail: sebol@bk.ru) Bolshakova Svetlana Evgenyevna – Candidate of Medical Sciences, Research Officer at the Laboratory of Pediatrics and Neuro physiology of Scientific Centre for Family Health and Human Reproduction Problems (664003, Irkutsk, ul. Timiryazeva, 16; e-mail: sebol@bk.ru) ул р , ; ) Bolshakova Svetlana Evgenyevna – Candidate of Medical Sciences, Research Officer at the Laboratory of Pediatrics and Neuro physiology of Scientific Centre for Family Health and Human Reproduction Problems (664003, Irkutsk, ul. Timiryazeva, 16; e-mail: sebol@bk.ru) Педиатрия Педиатрия 61 ACTA BIOMEDICA SCIENTIFICA, 2017, Vol. 2, N 4 Гомелля Марина Владимировна – доктор медицинских наук, доцент, научный сотрудник лаборатории педиатрии и кардиоваскулярной патологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (тел. (3952) 20- 73-67; e-mail: iphr@sbamsr.irk.ru) Гомелля Марина Владимировна – доктор медицинских наук, доцент, научный сотрудник лаборатории педиатрии и кардиоваскулярной патологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (тел. (3952) 20- 73-67; e-mail: iphr@sbamsr.irk.ru) ; p ) Gomellya Marina Vladimirovna – Doctor of Medical Sciences, Docent, Research Officer at the Laboratory of Pediatrics and Cardiovascular Pathology of Scientific Centre for Family Health and Human Reproduction Problems (tel. (3952) 20-73-67; e-mail: iphr@sbamsr.irk.ru) ; p ) Gomellya Marina Vladimirovna – Doctor of Medical Sciences, Docent, Research Officer at the Laboratory of Pediatrics and Cardiovascular Pathology of Scientific Centre for Family Health and Human Reproduction Problems (tel. ACTA BIOMEDICA SCIENTIFICA, 2017, Vol. 2, N 4 (3952) 20-73-67; e-mail: iphr@sbamsr.irk.ru) Баирова Татьяна Ананьевна – доктор медицинских наук, руководитель лаборатории персонализированной медицины ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Bairova Tatyana Ananyevna – Doctor of Medical Sciences, Head of the Laboratory of Personalized Medicine of Scientific Centre for Family Health and Human Reproduction Problems Баирова Татьяна Ананьевна – доктор медицинских наук, руководитель лаборатории персонализированной медицины ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Bairova Tatyana Ananyevna – Doctor of Medical Sciences, Head of the Laboratory of Personalized Medicine of Scientific Centre for Family Health and Human Reproduction Problems Бердина Ольга Николаевна – кандидат медицинских наук, ведущий научный сотрудник лаборатории педиатрии и ней рофизиологии ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» (e-mail: goodnight_84@mail.ru) Berdina Olga Nikolaevna – Candidate of Medical Sciences, Leading Research Officer at the Laboratory of Pediatrics and Neurophysiology of Scientific Centre for Family Health and Human Reproduction Problems (e-mail: goodnight_84@mail.ru) Бугун Ольга Витальевна – доктор медицинских наук, заместитель директора по клинической работе ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Bugun Olga Vitalyevna – Doctor of Medical Sciences, Deputy Director for Clinical Work of Scientific Centre for Family Health and льга Витальевна – доктор медицинских наук, заместитель директора по клинической работе ФГБНУ «Научн облем здоровья семьи и репродукции человека» Бугун Ольга Витальевна – доктор медицинских наук, заместитель директора по клинической работе ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Bugun Olga Vitalyevna – Doctor of Medical Sciences, Deputy Director for Clinical Work of Scientific Centre for Family Health and Human Reproduction Problems д р р р ду ц Vitalyevna – Doctor of Medical Sciences, Deputy Director for Clinical Work of Scientific Centre for Family Health and oduction Problems Михалевич Исай Моисеевич – кандидат геолого-минералогических наук, инженер ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Mikhalevich Isay Moiseevich – Candidate of Geological and Mineralogical Sciences, Engineer at Scientific Centre for Family Health and Human Reproduction Problems Рычкова Любовь Владимировна – доктор медицинских наук, профессор РАН, директор ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Rychkova Lyubov Vladimirovna Doctor of Medical Sciences Professor of the Russian Academy of Sciences Director of Scientific Рычкова Любовь Владимировна – доктор медицинских наук, профессор РАН, директор ФГБНУ «Научный центр проблем здоровья семьи и репродукции человека» Rychkova Lyubov Vladimirovna – Doctor of Medical Sciences, Professor of the Russian Academy of Sciences, Director of Scientific Centre for Family Health and Human Reproduction Problems 62 Pediatrics	26,913
https://openalex.org/W1728575383_2	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	7,848	14,721	Características de las empresas 14 Un estudio en profundidad de 21 empresas que usaban algún tipo de biotecnología agrícola en ocho países de la región, realizado por el IICA en 1990 (Jaffé, 1993, Jaffé, 1994), complementado con conclusiones de estudios de caso similares hechos en Brasil (Cerantola, 1991), permite caracterizar las empresas latinoamericanas de biotecnología en cuanto a sus recursos y estrategias gerenciales. grupo de empresas establecidas, productoras de insumos para la agricultura y la industria, están usando biotecnologías, dado que ellas afectan o afectarán directamente sus procesos productivos. Finalmente, existe un grupo de empresas agroindustriales que se están involucrando con la biotecnología debido a la amenaza o el potencial que ella representa para sus negocios en el mediano o largo plazo. Las empresas caen dentro de tres categorías, cada una de las cuales con su lógica particular. Las nuevas empresas de biotecnología fueron creadas con el propósito de explotar comercialmente alguna biotecnología. Un Las nuevas empresas de biotecnología son pequeñas y jóvenes empresas intensivas en tecnología, es decir, cuyo éxito o competitividad depende principalmente de su tecnología, que están en proceso de Cuadro 7. Algunas características de empresas latinoamericanas de biotecnología (% del total, 1991-92) País Tipo Propiedad NEB Establecida Local Extranjera Gobierno Argentina 32,4 67,5 89,1 10,8 0,0 Bolivia 85,7 14,2 28,5 0,0 71,4 Brasil 40,2 59,7 65,2 25,0 9,7 Chile 14,2 85,7 100,0 0,0 0,0 Colombia 30,7 69,2 76,9 7,6 15,3 Costa Rica 66,6 33,3 61,1 22,2 16,6 Ecuador 30,0 70,0 30,0 70,0 0,0 El Salvador 0,0 100,0 0,0 100,0 0,0 Guatemala 50,0 50,0 100,0 0,0 0,0 Honduras 0,0 100,0 0,0 0,0 100,0 México 38,2 61,7 67,6 26,4 5,8 Perú 28,5 71,4 100,0 0,0 0,0 Uruguay 38,1 61,9 80,9 19,0 0,0 Venezuela 68,5 34,4 75,8 3,4 20,6 Total 42,5 57,4 71,4 18,6 9,8 Fuente: IICA (1992 y 1993a) 18 consolidación. La falta de capital y de financiamiento de riesgo en la región las obliga a adoptar estrategias de crecimiento financieramente cautas y conservadoras, y a seguir planes de producción y de mercadeo oportunistas y altamente flexibles. En consecuencia, presentan estructuras flexibles, informales, pero sofisticadas, y culturas organizacionales que asignan un alto valor a su personal. Estas empresas, en contraste con las nuevas empresas de biotecnología del norte, que pasan años realizando solo IyD, producen y mercadean productos desde su creación, lo que significa que explotan biotecnologías más o menos maduras y desarrolladas. capacidades de investigación, que son aún muy limitadas. La trayectoria lógica de las empresas de insumos las llevará a una progresiva conversión en empresas de base tecnológica, a diferencia de las agroindustrias, para las cuales la biotecnología será menos central. La situación de la biotecnología agrícola El desarrollo de la biotecnología agrícola en ALC, aun cuando se han alcanzado ciertos niveles significativos en algunos países e industrias, es incipiente y débil, comparado con el de los países desarrollados. La inversión que se está haciendo en este campo en la región es insuficiente, especialmente la que hace el sector productivo. Los ejemplos más frecuentes de tecnologías explotadas son el cultivo de tejidos vegetales (propagación de frutales y ornamentales), las pruebas inmunológicas (estuches de diagnósticos veterinarios) y la producción de biopesticidas (bacterias, virus y hongos). Su enfoque de desarrollo tecnológico es de corto plazo y de carácter generalmente informal, dado que no pueden costear procesos de IyD largos. Muchas de las nuevas empresas de biotecnología han pasado ya por varias fases de desarrollo, comenzando por su constitución, su consolidación y finalmente expansión. Para que estas empresas sean viables en el mediano plazo deben superar ciertos umbrales financieros, organizacionales y tecnológicos. Los esfuerzos de IyD en biotecnología en la región, por lo general, son ineficientes y de una orientación inadecuada. Son dispersos, y demasiado ambiciosos para la infraestructura y personal disponibles, dependiendo principalmente de fondos gubernamentales. Uno de los aspectos mas débiles en la región es la bioingeniería o bioprocesamiento (Quintero, 1994). Solo un pequeño grupo de centros de investigación cuentan con la masa de recursos necesaria para una investigación de calidad y relevancia. La mayoría de los grupos son muy pocas personas acometiendo demasiados proyectos y actividades. Las empresas establecidas de insumos biológicos están apenas comenzando a incursionar en las nuevas tecnologías. Lo hacen de forma cautelosa, relacionándose para ello con centros de investigación, a través de contratos o de manera más informal (intercambios de investigadores, pasantías, colaboraciones informales). Algunas muy pocas tienen estrategias explícitas de adopción de biotecnologías, dedicándole a este proceso recursos significativos. Finalmente, las agroindustrias también estaban en una fase temprana de su involucramiento con las nuevas tecnologías. Las siguientes conclusiones de un extenso estudio realizado en México son válidas para toda la región, quizá con excepción de Cuba. Las limitaciones más importantes no son tanto la falta de recursos, como sostenían los investigadores, sino problemas más estructurales, como la falta de mecanismos para transferir resultados al sector productivo, estructuras académicas muy rígidas y falta de prioridades y políticas de investigación explícitas (Casas, 1993). La efectividad de la investigación en biotecnología es afectada negativamente por la falta de una clara idea del papel que la investigación juega en el desarrollo de la biotecnología a nivel nacional. Son muy comunes expectativas poco realistas en relación a las potencialidades de la biotecnología, así como desconocimiento o subestimación de las dificultades para desarrollar procesos y productos comerciales. Las empresas de insumos de base biológica y las agroindustrias establecidas inician, por lo general, sus actividades en biotecnología explorando que les puede ofrecer el nuevo campo, usualmente a través de sus relaciones con grupos de investigación nacionales. Eventualmente desarrollan sus propias 19 Es decir, muchos grupos de investigación buscan desarrollar biotecnología sin los necesarios recursos. significativo de investigación dentro de la industria, es decir, realizada por ella misma en sus propias facilidades. La metodología utilizada en la mayoría de los diagnósticos revisados no permite identificar la investigación realizada para la industria en los centros de investigación. Los pocos datos disponibles hacen pensar que este es un porcentaje pequeño del esfuerzo total, quizá alrededor del 10%. Un importante volumen de recursos está disperso en un número grande de instituciones, que no cuentan con la masa crítica necesaria para hacer buena ciencia y mucho menos, desarrollar productos o procesos comerciales. Pero, al menos, existe un grupo de personas con capacidad para comprender y monitorear los desarrollos internacionales en este campo. Esto es un elemento importante para la educación y formación profesional más general. Pero sin duda, por lo menos Brasil, Argentina, México y Cuba, y, en menor medida Chile, han desarrollado una base científica para un eventual desarrollo más amplio de la biotecnología agrícola. Es de destacar que no existe un esfuerzo 20 Estrategias y políticas de desarrollo de la biotecnología Aun cuando hay una cierta espontaneidad en el desarrollo de la biotecnología, particularmente en sus fases tempranas donde la iniciativa individual de científicos es importante, no puede pensarse en la creación y consolidación de capacidades productivas en este campo sin planes deliberados de empresas y productores. El éxito de los mismos dependerá de las estrategias que se sigan, así como de la existencia de un conjunto de políticas gubernamentales que ofrezcan el marco legal, regulatorio y, en general, económico, adecuado para el desarrollo de la biotecnología. potencial de la biotecnología para el mejoramiento de la calidad y productividad de estos sectores y se lograría mantener o mejorar su competitividad. Los requerimientos para una incorporación rápida y eficiente de las nuevas tecnologías en actividades productivas e industrias existentes definen las estrategias para el logro de este objetivo. Usar estas tecnologías adecuadamente exige de un mínimo de conocimiento sobre ellas, lo que implica disponer de una capacidad para entender y seguir los desarrollos internacionales más importantes para cada sector o empresa y eventualmente una capacidad para adaptarlas a los requerimientos y condiciones locales. Esto es válido aun en los casos en que el poseedor de la tecnología ofrezca asistencia técnica para su uso. Las estrategias empresariales y gubernamentales de la biotecnología son guías indispensables para la orientación de las acciones en función de los determinantes internacionales y nacionales del desarrollo de este campo. La condición de seguidores, es decir, la existencia de líderes internacionales en biotecnología y las limitaciones de recursos y capacidades, tanto a nivel de empresa como nacional, son los determinantes más importantes en este sentido. En el plano nacional ello significa disponer de un grupo más o menos grande de personas versadas en las nuevas tecnologías genéricas o básicas, que usualmente son investigadores en alguna institución gubernamental. Las empresas o grupos de productores, por su lado, también deben contar con un mínimo de personal conocedor de la biotecnología, capaz de relacionarse adecuadamente con proveedores o investigadores. De particular importancia será la existencia de un marco legal y regulatorio nacional que permita una rápida difusión de las tecnologías al país y el acceso de las empresas nacionales a las mismas. Las regulaciones de bioseguridad y normativas de propiedad intelectual adecuadas son los dos elementos claves de este marco. Estrategias nacionales y empresariales Los países en desarrollo, así como muchas empresas en los mismos países desarrollados, están rezagados respecto a las nuevas tecnologías. Las empresas y países líderes llevan muchos años de ventaja, en términos de inversiones hechas, tecnologías desarrolladas y aprendizajes realizados. Sin embargo, los rezagados igual se verán afectados por los impactos de las nuevas tecnologías. Entre los más directos e inmediatos está la disponibilidad de nuevos insumos biológicos para muchos sectores tradicionales, que van a modificar sus patrones de calidad y de productividad y, en consecuencia, su acceso a mercados y su competitividad. Por ello, la primera prioridad para todo aquel que realiza actividades productivas de base biológica debería ser incorporar las nuevas tecnologías vía insumos en el momento que estos estén disponibles. Así se aprovecharía el Esta estrategia de “usuario de biotecnología” es la apropiada para los países de la región que tengan importantes industrias agrícolas y agroindustriales, pero que carezcan de industrias productoras de insumos biológicos. Países que tengan estas últimas se ven forzados a acciones más proactivas y ambiciosas si quieren mantenerlas. Las nuevas tecnologías amenazan directamente estas empresas, pues tienen el potencial de sustituir 21 sus procesos y productos tradicionales. Esto las obliga a incorporar las biotecnologías como tales, es decir, no como insumos sino como nuevos procesos y procedimientos. Leopold, 1991). Una estrategia tecnológicamente agresiva de este tipo, exige un esfuerzo de IyD importante por parte de la empresa. Pero hay opciones alternativas para disponer de tecnologías requeridas como son la compra, el licenciamiento, la asociación estratégica, los emprendimientos conjuntos, etc., como lo muestran las estrategias de las compañías líderes en biotecnología. El objetivo en una primera instancia en el plano nacional en este caso es posicionar la industria local productora de insumos biológicos agrícolas e industriales para una eventual incorporación de las nuevas tecnologías. Tarde o temprano estas compañías tendrán necesariamente que reconvertirse hacia empresas basadas en biotecnologías. También existirán oportunidades para la creación de nuevas empresas o la entrada a nuevos mercados por parte de empresas existentes (Jaffé, 1993). La decisión de seguir una estrategia de líder tecnológico es fundamentalmente de la empresa. Pero puede ser apoyada o inclusive impulsada por una voluntad nacional, expresada en estrategias y políticas gubernamentales, de lograr este liderazgo. Algunos pocos países en desarrollo pueden tener los recursos y generar la suficiente voluntad política para alcanzar a algunos líderes a través de un proceso más o menos autónomo de desarrollo de la biotecnología. El conjunto de acciones con estos objetivos conformarían una estrategia de “seguidor inteligente” (Solleiro, 1995), que permitiría a las empresas locales competir en sus países aprovechando ventajas locales y eventualmente incluso a nivel internacional, con base en factores no tecnológicos (mercadeo, costos, etc.). Existen, de hecho, algunos pocos ejemplos de empresas farmacéuticas y de fermentaciones latinoamericanas que lo hacen en la actualidad exitosamente. Dada la relativa debilidad de la base científica local, esta competitividad probablemente no esté tanto basada en nuevos productos o procesos, sino en una eficiente manufactura o comercialización. El papel del gobierno El desarrollo de la biotecnología en cualquier país es el producto de un complejo conjunto de acciones de científicos, directivos de instituciones de IyD, empresarios, productores, financistas y funcionarios gubernamentales. Las acciones gubernamentales son de especial importancia, pues es usualmente esta instancia que puede poner en marcha un proceso de este tipo en aquellos países o sectores donde no existan suficientes científicos y empresarios para hacerlo por su cuenta. Es decir, uno de los roles del gobierno es la creación y mantenimiento de la masa crítica científica y profesional necesaria para un proceso de desarrollo de la biotecnología. No puede descartarse a priori la posibilidad de competir internacionalmente también en términos tecnológicos. Alcanzar a los líderes es posible pero difícil. Los países relativamente más avanzados, algunas veces clasificados como semi-industrializados, que cuentan con sólidas bases científicas en los campos relevantes, pudieran aprovechar las oportunidades ofrecidas por el cambio de la base tecnológica en una determinada industria, un evento siempre turbulento por su alta incertidumbre intrínseca, la inercia del éxito del pasado, etc., para alcanzar nuevas posiciones competitivas o entrar en nuevos mercados. Este salto tecnológico requiere de muy buena programación en el tiempo para aprovechar la ventana de oportunidad entre el momento cuando las incertidumbres y riesgos aún son muy altos y cuando las barreras de entrada ya son altas debido a la consolidación de la posición de los líderes (Sercovitch y Como se ha insistido repetidamente, es necesario un cierto nivel de capacidades científicas y tecnológicas locales para el desarrollo de la biotecnología. En ciencia se requieren capacidades en biología molecular y celular, así como en microbiología aplicada. Estas debería complementarse con capacidades en ingenierías relacionadas con el campo, especialmente bioprocesamiento. Un cierto dominio de las biotecnologías genéricas o básicas principales, tales como cultivo de tejidos, técnicas inmunológicas, ingeniería genética, es esencial. 22 Una vez iniciado el proceso, su éxito va a depender en gran medida de la eficiencia global del mismo, que depende de la efectiva solución de la multiplicidad de requerimientos técnicos, económicos y comerciales de la investigación y producción en biotecnología. Financiamiento, provisión de insumos, reactivos y equipos, servicios de apoyo, información técnica y comercial, mantenimiento, entrenamiento, normas y regulaciones, son ejemplos de algunos de estos requerimientos. Es necesario un cierto nivel de coordinación entre esta multiplicidad de actores, que permita una mejor orientación de los esfuerzos individuales, pero también la identificación y superación de cuellos de botella. Esta coordinación se ve facilitada por una visión estratégica de mediano y largo plazo más o menos compartida por los distintos actores. la iniciativa privada porque no son atractivos comercialmente o porque su importancia estratégica haga que el Estado los asuma exclusivamente. Enfermedades de poblaciones de bajo nivel económico o de pequeños grupos, problemas de salud pública, tecnologías para pequeños productores agrícolas pobres, aplicaciones ambientales o militares, son algunos ejemplos de problemas de este tipo. Estos solo pueden ser asumidos por el Estado, lo que dependerá de la política gubernamental, y de la influencia política de los sectores interesados. En resumen, el papel del gobierno en un proceso de desarrollo de la biotecnología puede, entonces, estar dirigido hacia los siguientes objetivos: La coordinación, en principio, debería generarla el propio mercado, aun en forma informal, como se evidencia en algunos de los países líderes. Pero en procesos de desarrollo de la biotecnología incipientes y débiles la misma debe ser facilitada y posibilitada por la acción gubernamental que, por lo general, es la única con la suficiente capacidad de convocatoria y neutralidad para hacerlo. Más aún, esta acción gubernamental puede catalizar e impulsar el proceso, imprimiéndole el dinamismo y la orientación que los actores individualmente no pueden lograr. El desarrollo de la biotecnología requiere de una infraestructura que supla sus requerimientos, así como de un marco legal y regulatorio adecuado que oriente su desarrollo. Muchos de los servicios requeridos solo pueden ser asumidos por la iniciativa privada en estadios más avanzados del proceso, por razones de escalas y de falta de capacidades básicas, por lo que se justifica la acción gubernamental para suplirlos temporalmente. La creación y administración del marco legal y regulatorio es responsabilidad del Estado. Sin embargo, algunos aspectos de este marco pueden ser asumidos por el sector privado, como lo demuestra el caso del control de calidad de semillas, que en algunos países es realizado por la propia industria. Ø Creación y mantenimiento de capacidades científicas básicas y de la formación de los recursos humanos requeridos. Ø Creación y administración del marco legal y regulatorio requeridos para salvaguardar los intereses públicos y facilitar y orientar la inversión y el comercio. Ø Proveer o catalizar la provisión de los servicios técnicos, financieros y comerciales requeridos. Ø Realizar la IyD y la producción en aquellos problemas o áreas que por razones de escala, capacidad de compra o por interés estratégico no van a ser asumidos por la iniciativa privada. Los objetivos dependen del nivel de desarrollo alcanzado en el país. Es decir, la intervención gubernamental debe tomar en cuenta la fase en la que se encuentra el país en términos de desarrollo biotecnológico. Muchos países han implementado programas explícitamente dirigidos a desarrollar la biotecnología, desde comienzos de la década de los años ochenta. En otros se identifican acciones más aisladas de organizaciones gubernamentales, generalmente centros de investigación, para desarrollar capacidades en biotecnología. Siempre van a existir problemas, potencialmente superables mediante la biotecnología, que no van a ser abordados por 23 Los programas de desarrollo de la biotecnologíaLos programas de desarrollo de la biotecnolog’a sido los más comunes en la región. A medida que los países avanzan en la consolidación y expansión de sus capacidades, estos programas se hacen más especializados y relativamente menos importantes frente a programas con otros objetivos. Surgen como iniciativa y por la presión de científicos o grupos de ellos influyentes, canalizada a través de los organismos nacionales de promoción o desarrollo científico y tecnológico. Una forma de ellos combina el financiamiento de IyD, con el desarrollo de infraestructura y el entrenamiento de recursos humanos. Argentina, Brasil, Colombia, Costa Rica y Venezuela son países que han tenido o tienen este tipo de programas (Cuadro 8). Algunos de estos programas han sido financiados por el BID (Costa Rica y Venezuela) y el Banco Mundial (Brasil). El campo de las ingenierías bioquímicas no ha sido incluido en este tipo de programas (Aguilera y col., 1994). Los programas de desarrollo de la biotecnología generalmente son acometidos por organismos gubernamentales, idealmente dentro de estrategias nacionales o sectoriales de desarrollo de la biotecnología. Pueden ser llevados a cabo por ministerios sectoriales, centros de investigación y organismos de promoción de ciencia y tecnología, con fondos propios o de organismos de financiamiento multilaterales. Una forma de darle coherencia y visibilidad a acciones para el desarrollo de la biotecnología es estructurar un programa nacional de biotecnología. En esta figura pueden hacerse confluir las acciones y estrategias de diversas organizaciones, coordinadas y dirigidas por alguna instancia o cuerpo representativo de los actores más importantes del proceso. La formación de recursos humanos en biotecnología generalmente se ha realizado en el exterior. El énfasis ha sido en la formación a nivel de doctorado, y no siempre se ha cuidado la capacidad de absorción del personal formado por parte de los sistemas científicos nacionales, con el resultado que algunas de las personas formadas se queden en el exterior. No se ha dado la suficiente importancia a la formación más operativa e instrumental, es decir, a nivel de maestría, de entrenamiento y de formación técnica. En algunos países de América Latina existen o existieron estructuras de este tipo, denominadas comisiones o comités nacionales de biotecnología. Para que una comisión de este tipo funcione adecuadamente se requiere de una visión estratégica compartida por los diferentes interesados acerca del potencial papel de la biotecnología en la economía y la ciencia en el país. Los programas dirigidos al desarrollo y fortalecimiento de capacidades científicas han 24 Cuadro 8. Ejemplos de programas gubernamentales de apoyo directo a la biotecnología en América Latina País Nombre y fecha Organismo responsable (Financiador) Componentes Inversión (millones US$) Argentina Programa Nacional de Biotecnología (1986) Secretaría de Ciencia y Técnica Promoción y financiamiento de IyD 3,8 Brasil Programa Nacional de Biotecnología (1981) CNPq y FINEP Financiamiento de IyD 3,3 PADCT/ Biotecnología (1984) Ministerio de CyT (Banco Mundial) IyD Formación de personal Infraestructura 12,9 Polos de biotecnología Ministerio de CyT Infraestructura y organización común para empresas Chile Comité Nacional de Biotecnología (1983) CONICYT Coordinación y promoción de IyD y formación de recursos humanos Colombia Programa Nacional de Biotecnología (1984) COLCIENCIAS Financiamiento de IyD planificación y coordinación Venezuela Programa Nacional de Ingeniería Genética y Biotecnología (1986) CONICIT Financiamiento de IyD 0,5 Programa Nuevas Tecnologías (1992) CONICIT (BID) IyD Recursos humanos Infraestructura 30,0 Fuente: Con base en datos de Correa (1992) e información directa El componente de infraestructura ha incluido la adquisición de equipos y la construcción de facilidades de investigación, frecuentemente laboratorios dentro de institutos de investigación o universidades. Otra alternativa, más enfocada, ha sido la creación de institutos de biotecnología dentro de universidades o centros de investigación aplicada. Ejemplos destacados de centros creados en los últimos quince años en la región son el Instituto de Biotecnología de la Universidad Autónoma de México, la Unidad de Irapuato del Centro de Investigación y Estudios Avanzados (CINVESTAV) en México, el Instituto de Ingeniería Genética y Biotecnología de Cuba, el Centro Nacional de Investigación en Recursos Genéticos y Biotecnología (CENARGEN) de la Empresa Brasilera de Investigación Agropecuaria (EMBRAPA) en Brasil, el Centro de Biología Molecular del Instituto Nacional de Tecnología Agropecuaria (INTA) y el Centro de Ingeniería Genética del Centro Nacional de Ingeniería Genética (CONICET), ambos en Argentina. Han 25 demostrado ser una forma adecuada de crear una masa crítica, que a su vez dinamiza el desarrollo de la biotecnología más allá de su entorno inmediato, a través de la formación de recursos humanos y la visibilización de la biotecnología y su importancia. provisión de capital de riesgo, facilitar la creación de empresas, el relacionamiento de los diferentes actores entre sí y el desarrollo de una visión estratégica compartida sobre el papel de la biotecnología en la agricultura y la economía. Programas de fortalecimiento de las ciencias básicas, aun cuando no tienen el objetivo explícito de desarrollar la biotecnología, pueden ser de gran importancia si incluyen las disciplinas bases de la biotecnología. Este ha sido el caso de los programas PEDECIBA en Uruguay, y el PDCT en el Brasil, financiado por el Banco Mundial. Solo Cuba tiene un programa nacional, ambicioso e integrado, de desarrollo de una industria de biotecnología. Con base en las capacidades de algunos pocos institutos de investigación se está desarrollando una industria de exportación de productos biotecnológicos, así como de servicios médicos y agrícolas. Programas para desarrollar la infraestructura general requerida por el desarrollo de la biotecnología, como tal, no existen en la región. Sin embargo, varios de los países más desarrollados cuentan con programas de fomento de la innovación y el desarrollo tecnológico en general, que obviamente también benefician a la biotecnología. Los apoyos a la investigación en la industria y a la creación y expansión de empresas en general, han jugado un papel importante en la adopción de biotecnologías por parte de la industria (Jaffé, 1993). Las empresas de biotecnología han aprovechado eficientemente estas facilidades disponibles en sus países. Sin duda, la más importante para ellas es la existencia de grupos de investigación de alto nivel a los que recurrir para asistencia técnica, información, capacitación de personal e IyD. Marco legal y regulatorio Las iniciativas de creación o fortalecimiento del marco legal y regulatorio requerido por la biotecnología que se han dado en la región generalmente no han surgido de programas de desarrollo de ella, sino de necesidades e intereses más particulares, como son los de la industria de semillas o de fitomejoramiento, las presiones de algunos países o las exigencias de acuerdos multilaterales. De esta forma se han impulsado la modificación de leyes de propiedad intelectual y la introducción de regulaciones de bioseguridad en algunos países. Propiedad intelectual La creciente importancia económica de la innovación ha impulsado fuertemente el fortalecimiento de la protección de la propiedad intelectual a nivel internacional, con el resultado que este tema ha sido incluido en la agenda de diversas negociaciones de libre comercio, destacándose la Ronda Uruguay del Acuerdo General sobre Tarifas y Comercio (GATT) y el Tratado de Libre Comercio (TLC). Por otro lado, la biotecnología permite la apropiación privada de tecnologías e información que tradicionalmente han estado en el ámbito público. La conjunción de estas dos tendencias ha impulsado la extensión de la protección de la propiedad intelectual a nuevas áreas, tales como la agricultura, los seres vivos y la información biológica en general. En este contexto, destacan programas diseñados para suplir alguna necesidad específica del desarrollo biotecnológico a nivel comercial. Una iniciativa interesante en este sentido es el proyecto de la Universidad Federal de Río de Janeiro (BIORIO), que ofrece servicios de incubación de empresas y de parque industrial especializados en biotecnología (Paes de Carvalho, 1995). Una política dirigida al desarrollo comercial de la biotecnología debería ser selectiva en términos de industrias o sectores a apoyar, para permitir un adecuado foco y masa crítica de esfuerzos, dada la limitación de recursos. A nivel macro es importante contar con un ambiente adecuado para la inversión extranjera y la transferencia de tecnología desde países más avanzados, así como para la innovación. Tal política enfatizaría la El ajuste de las legislaciones nacionales de propiedad intelectual a alguna norma internacionalmente aceptada es visto por algunos como una importante condición para 26 el acceso a tecnología avanzada, así como para el desarrollo de capacidades de innovación locales. En respuesta a fuerte presiones internacionales, muchos países latinoamericanos han revisado recientemente sus legislaciones de propiedad intelectual. De esta forma, leyes de patentes recientes en México, el Pacto Andino, Chile y Argentina introdujeron la protección de microorganismos y los procesos basados en ellos, así como de farmacéuticos y alimentos. El patentamiento de otros seres vivos aún no se permite. Estos cambios son demasiado recientes como para haber tenido algún efecto sobre el desarrollo de la biotecnología en la región. avanzadas, de especial importancia para las industrias de flores y frutas. Los intentos de proteger y apropiar recursos biológicos avanzaron en forma importante con la aprobación de la Convención de Protección de la Diversidad Biológica y su rápida ratificación por un gran número de países. En ella se establece la soberanía nacional sobre los recursos genéticos, lo cual está comenzando a tener un impacto importante sobre la investigación agrícola, y en particular el fitomejoramiento. Algunos institutos de investigación agropecuaria (INIAs) en América Latina están en el proceso de establecer normativas de acceso e intercambio de germoplasma, en forma similar a las prácticas que se están introduciendo en los centros internacionales. Los países del Pacto Andino están desarrollando una política común de acceso a los recursos genéticos. Otra forma de propiedad intelectual que ha tenido un desarrollo importante en la región recientemente ha sido los derechos de obtentores vegetales. Como instrumento de protección de los derechos de los fitomejoradores tienen importancia en todos los países con un sector agrícola de peso, y más particularmente con una industria de semillas y de fitomejoramiento importante. Argentina, Chile y Uruguay cuentan con este tipo de legislación desde hace 20 años aproximadamente, aun cuando no se implementaron efectivamente desde los comienzos de los años noventa. En este mismo período se introducen este tipo de normativas en México y los países del Pacto Andino. Otros países (Brasil, Costa Rica) están en el proceso de hacerlo (Jaffé y van Wijk, 1995). Bioseguridad La bioseguridad, es decir, las regulaciones para reducir los riesgos de la biotecnología para la salud, los sistemas productivos y el medio ambiente, son un elemento indispensable de cualquier política de desarrollo de la biotecnología (Trigo y Jaffé, 1991). Además de la obligación moral y práctica de resguardar los intereses públicos y privados, buscan facilitar el comercio y la transferencia de tecnología mediante el establecimiento de estándares y prácticas internacionalmente aceptadas. Un estudio reciente de la Universidad de Amsterdam y el IICA determinó que los impactos de estas legislaciones aún eran pocos en los países que han contado con ellas por cierto tiempo (Jaffé y van Wijk, 1995). Aún así, se detecta un impacto positivo sobre el fortalecimiento del fitomejoramiento local de variedades de polinización abierta. Adicionalmente son un elemento importante para el acceso de la industria local a variedades Algunos de los países relativamente más desarrollados de la región comenzaron a tratar el problema de la bioseguridad. Brasil, México y Cuba, por ejemplo, en la década pasada establecieron guías nacionales para el uso de técnicas de ADN recombinante. El énfasis ha estado en la regulación de la liberación en el medio ambiente de organismos transgénicos. 27 Latina Cuadro 10. Programas regionales de cooperación técnica en biotecnología País 198 199 199 199 199 Total 9 0 1 2 3 Fecha Nombre Argentina Agencia 0 4 Ambito 6 10 20Inversión (US$) 1988-93 Programa Belice Regional de Biotecnología Bolivia BIOLAC Chile PNUD/UNESCO 0 0 0 ONUDI 0 0 2 Universidad de las Naciones 0 Unidas 0 2 Regional 3 ? 35.000.000 0 0 Regional 1 1 2 150.000-200.000 4por año 1990- REDBIO Costa Rica FAO 0 Regional 4 ? 560.000 por año 1988-93 Programa Rep. Dominicana Andino de Corporación 0 0 Andina1de Biotecnología Fomento México 0 1 1 Políticas para IICA Total 0 1 11 Biotecnología Agrícola Fuente: Base de datos IICA Biotecnología OEA Región ? Andina ? 12.000.000 4 ? Regional 18 11 6 800.000 42 Regional 300.000 por año Programa Biotecnología Cono Sur 120.000 por año 1988- 1988-1994 19881992- 0 0 1 Programa Cooperativo de Investigación Agrícola del Cono Sur (PROCISUR)/ IICA Como respuesta al interés de empresas transnacionales de ensayar plantas transgénicas, es decir, obtenidas por ingeniería genética a nivel de campo, México, Costa Rica, Bolivia, Argentina y Chile crearon comités para evaluar y supervisar estos ensayos, basados en regulaciones fitosanitarias existentes. Un número significativo de ensayos se han realizado en la región, como se muestra en el cuadro 9. Esto es el resultado de la respuesta flexible y oportuna de algunos países a los requerimientos de bioseguridad. Brasil destaca como el único país que no ha podido desarrollar una respuesta de este tipo, ya que seleccionó la vía legislativa para introducir regulaciones de bioseguridad (Sant’Ana, 1992). La cooperación internacional apoyó en forma determinante la temprana difusión de la biotecnología a la región y particularmente del reconocimiento de su importancia. También aportó y aporta en la actualidad recursos críticos para el desarrollo de la biotecnología, de especial importancia para los países menos desarrollados. En esta tarea participaron las agencias de cooperación técnica internacionales y regionales, los centros de investigación agrícola y de biotecnología internacionales y algunas agencias de cooperación nacionales y privadas. En la última década varias agencias internacionales o regionales han desarrollado programas de cooperación técnica específicamente dirigidos al desarrollo de la biotecnología (Cuadro 10). El Programa Regional de Biotecnología de las Naciones Unidas (PNUD/UNESCO/ONUDI) fue el pionero de los mismos, aportando fondos para la realización de proyectos cooperativos entre países, cuyos resultados, más que productos o procesos específicos, fueron la difusión de un conjunto de técnicas básicas a un número significativo de países e instituciones de investigación. También fomentó la Un factor importante de esta respuesta efectiva ha sido la existencia de guías de bioseguridad regionales, producidas por el IICA y la Organización Panamericana de la Salud (OPS) (IICA y PAHO, 1988; 1991), así como la asistencia técnica ofrecida por la Oficina de Protección Agrícola del Departamento de Agricultura de los Estados Unidos (USDAAPHIS). La cooperación internacional 28 constitución de comisiones nacionales de biotecnología, que fueron un ensayo interesante de coordinación nacional de los esfuerzos de promoción de la nueva tecnología. Centro de Investigación de la Agricultura Tropical y el Centro Agronómico Tropical de Investigación y Enseñanza (CATIE), a nivel de Centroamérica, han sido y son factores importantes en el desarrollo de la biotecnología, por su labor en la difusión de técnicas claves a través del entrenamiento y formación de personal. La amplia difusión de la técnica de cultivo de tejidos en la región a comienzos de la década de los ochenta, por ejemplo, se debe principalmente a la labor del CIP y de sus redes regionales de investigación en papa. La Unidad de Biotecnología del CIAT, además de ser uno de los grupos de investigación más avanzados en la región, ha apoyado la creación y fortalecimiento de capacidades nacionales en biotecnología mediante un intenso intercambio y entrenamiento de investigadores. Otras iniciativas también dirigidas a crear capacidades básicas son el programa BIOLAC de la Universidad de las Naciones Unidas, dedicado al entrenamiento de personal y la creación de redes regionales en biotecnología vegetal por parte de la Organización de las Naciones Unidas para la Agricultura y la Alimentación (FAO). Dentro de los programas subregionales de investigación agrícola se han creado programas de biotecnología para realizar proyectos conjuntos y transferir capacidades e información. El Programa Cooperativo de Investigación Agrícola del Cono Sur (PROCISUR) es el ejemplo más destacado de los mismos. Diversas redes de investigación internacionales en biotecnología, apoyadas por agencias nacionales de países desarrollados o por fundaciones privadas, han servido igualmente para desarrollar capacidades nacionales, aun cuando no tienen como objetivo la cooperación técnica en sí sino la superación de problemas específicos de importancia para los países en vías de desarrollo. Esto frecuentemente lleva a la concentración de las actividades en los países líderes donde se encuentran las capacidades científicas. El único programa especializado en el apoyo a la definición e implementación de políticas en biotecnología fue el del IICA. Este programa se concentró en el tema de la bioseguridad, en el que se ha logrado un significativo avance en la región, en comparación con otras regiones del mundo. Los centros de investigación agrícola internacionales y regionales, y en particular el Centro Internacional de la Papa (CIP), el 29 Aspectos institucionales de la IyD en biotecnología agrícola La investigación y desarrollo en biotecnología tiene una serie de requerimientos técnicos y organizacionales específicos derivados de las características de este campo. En ella se combinan varias disciplinas y su objetivo final es eminentemente práctico, lo que la hace compleja en cuanto a su organización, dotación de recursos humanos y gerencia. La creación de grupos de IyD en biotecnología, bien en instituciones académicas, bien en organizaciones de investigación aplicada o de desarrollo experimental, es un objetivo que requiere ser cuidadosamente planificado, exigencia que frecuentemente se ha obviado, con consecuencias negativas sobre la consolidación y éxito posterior de las iniciativas. La subestimación de los recursos requeridos y de las dificultades gerenciales asociadas a la IyD en este campo ha sido común, una de las razones de la baja efectividad en el uso de los recursos invertidos en ella en la región. a su alcance, precisión y costos. Estas diferencias se ilustran en los ejemplos presentados en el cuadro 11, que contiene estimaciones de los recursos humanos y de la inversión requerida para desarrollar investigación con base en varias de las áreas más importantes de la biotecnología, realizadas para 1990. Estas estimaciones, aun cuando son aproximaciones para un país desarrollado (Canadá), dan una idea de la masa mínima de recursos requeridos para una IyD efectiva en varias biotecnologías genéricas o básicas. Aun cuando puede realizarse IyD en cada una de estas técnicas o servicios por separado, no puede obtenerse un producto final sin contar con varias de ellas simultáneamente, en forma ideal dentro de una misma institución. Por ejemplo, se hace poco con transformar genéticamente una bacteria o una planta si no se la puede cultivar en una escala significativa luego. Un laboratorio de cultivo de tejidos debería estar asociado con posibilidades de evaluar las plantas obtenidas en el campo. Por otro lado, todos los laboratorios o servicios en biotecnología requieren de un mínimo de apoyo en química analítica, cuyos costos y complejidades pueden variar enormemente, dependiendo de las técnicas requeridas. La información recogida en Colombia, El Salvador y Uruguay permite analizar la problemática organizacional y gerencial del desarrollo de capacidades científicas y tecnológicas en biotecnología y las distintas opciones que se ofrecen para organizar y planificar la IyD en este campo. Requerimientos técnicos y económicos Las estimaciones anteriores demuestran que, aun cuando montar un laboratorio de biotecnología es relativamente barato comparado con otras áreas de la ciencia y tecnología, no son pequeños los recursos requeridos. En consecuencia, la posibilidad de realizar esta inversión y de mantener el flujo de recursos requeridos en el tiempo para su funcionamiento es una consideración esencial en cualquier iniciativa para desarrollar IyD en biotecnología. La complejidad y costos de la IyD en biotecnología varían según el tipo de técnicas utilizadas. Por lo general, las técnicas celulares son menos complejas y exigen menos recursos que las técnicas moleculares. Sin embargo, se trata de un campo dinámico y en plena evolución, caracterizado por la rápida evolución de técnicas y equipos. Algunas son sustituidas por otras más precisas o baratas y otras son automatizadas o mejoradas en cuanto 30 Cuadro 11. Recursos requeridos para IyD en biotecnología (1990) Servicios o técnicas Personal Inversión en equipos (US$) Gastos de funcionamiento (US$) Ingeniería genética de microorganismos 3 PhD, 3 asistentes, 6 estudiantes 142.700 20.000 Escalamiento y salidas 2 PhD, 2 asistentes, 4 estudiantes 150.000 20.000 Cultivo de tejidos vegetal 1 Asistente, ayudantes Experimentación con plantas 1 Asistente, personal de apoyo Clonación de embriones animales 1 Asistente, personal de apoyo 85.000 Fuente: IICA (1993b) Organización Agrícolas (INIA) del Uruguay se creó una Unidad de Biotecnología con más capacidades y recursos, pero también con la misión básica de prestar servicios a los programas de investigación del instituto. Cuenta con cinco investigadores, uno de ellos a nivel de doctorado y uno a nivel de maestría, que realizan investigación para y en coordinación con varios de los programas, los cuales aprueban y financian sus proyectos. Aun los proyectos gestionados por la Unidad ante agencias de financiamiento externo son concertados con los programas. Realiza investigaciones de alta complejidad en el área de la biotecnología molecular y celular. La organización de las capacidades en biotecnología debe partir del carácter multidisciplinario de la IyD en este campo y de su carácter de servicio para otras líneas de actividad. El desarrollo de capacidades de biotecnología en organizaciones académicas o de investigación puede ser realizado de varias formas, como lo muestran los distintos casos examinados en los países analizados. Una primera opción es introducir el uso de las nuevas tecnologías en programas de investigación existentes y en sus correspondientes estructuras organizacionales, creando una unidad especial para ello. Por ejemplo, en el Centro de Tecnología Agrícola (CENTA) de El Salvador se estableció un pequeño laboratorio de cultivo de tejidos dentro del Programa de Recursos Genéticos, con dos investigadores sin posgrado, que presta servicios de termoterapia y propagación invitro. Sus capacidades se limitan a esta técnica, por lo que sus alcances son limitados. De manera similar, la Fundación de Servicios al Desarrollo (FUSADES), una fundación privada de asistencia técnica y servicios para la agricultura de El Salvador, tiene un laboratorio de diagnóstico y un laboratorio de cultivo de tejidos, que prestan servicios por demanda. El Instituto Nacional de Investigaciones Agrícolas de Colombia, CORPOICA, decidió organizar una capacidad en biotecnología más autónoma e independiente, pero que igualmente se pretende esté estrechamente relacionada con los programas de investigación operativa y regional. Dentro de la Subdirección de Investigación Estratégica se crearon un programa de biotecnología vegetal, y otro de biotecnología animal, que en la actualidad cuentan con 19 investigadores, 7 de ellos con nivel de doctorado. Cada programa tiene su propio presupuesto, que complementa con recursos externos de diversas fuentes. En el caso de instituciones que cuenten con capacidades en biotecnología en distintas dependencias, caso típico de muchas En el Instituto Nacional de Investigaciones 31 universidades donde hay grupos de biotecnología en medicina, agronomía, biología, farmacia, ingeniería, etc., se requiere instancias de coordinación y orientación de los esfuerzo. Este problema ha sido resuelto en la Universidad Nacional de Colombia mediante la creación de un Instituto de Biotecnología “virtual”, que mediante la estructuración de proyectos de investigación, de cursos conjuntos y el ofrecimiento de una infraestructura de uso común agrupa y coordina los investigadores en biotecnología de las distintas facultades. Cuenta solo con un pequeño grupo de personal propio. veterinarios en biotecnología, mediante entrenamientos o estudios de posgrado. Esta estrategia es factible en el caso del uso de técnicas relativamente menos complejas. Ha sido seguida por el CENTA, que tiene dos ingenieros agrónomos trabajando en su laboratorio de cultivo de tejidos; FUSADES, que tiene un ingeniero con una maestría encargado del laboratorio de cultivo de tejidos; el INIA, que tiene tres ingenieros a nivel de asistentes de investigación; y CORPOICA, que tiene igualmente agrónomos a nivel de asistentes en el Programa de Biotecnología Vegetal y médicos veterinarios en el de Biotecnología Animal. Un problema que se evidencia en varias instituciones es la falta de capacidades complementarias o adicionales para el desarrollo de productos finales dentro de la organización. Así, varias instituciones están desarrollando plantas transgénicas, pero no cuentan con programas de fitomejoramiento tradicionales ni con facilidades para ensayar las plantas en el campo. Ello obliga a establecer estrechas relaciones con grupos e instituciones que cuentan con estas facilidades y capacidades, que deberían establecerse idealmente desde el comienzo de los proyectos de biotecnología. Para desarrollar actividades más complejas se requiere un personal de nivel de doctorado y con una sólida formación en ciencias biológicas. Este tipo de personal normalmente no está disponible en los institutos de investigación agrícola por lo que debe ser reclutado especialmente. Este ha sido el caso del INIA, que reclutó al Jefe de su Unidad de Biotecnología en un instituto de investigación básica, y CORPOICA, que contrató a varios biólogos a nivel de doctorado y maestría para su programa de biotecnología vegetal. En el caso de los médicos veterinarios es más fácil su formación a nivel de doctorado en biotecnología, como lo muestra la experiencia del Programa de Biotecnología Animal de CORPOICA. Recursos humanos El personal especializado en biotecnología es, al menos, de dos tipos. Uno es el que trabaja en la modificación de los organismos y de sus productos, lo que exige una formación en biología celular o molecular, y otro que trabaja en la obtención o utilización de estos organismos y sus productos en diferentes escalas, por lo que tiene una formación de ingeniero (fitomejorador, ingeniero bioquímico, etc.). Idealmente se requiere de ambos tipos para desarrollar programas de biotecnología que resulten en productos o procesos. Este hecho es un primer elemento determinante de las estrategias de recursos humanos de una organización que quiera desarrollar capacidades integrales en biotecnología. Además de contar con personal especializado en biotecnología, es importante que otros investigadores tengan un conocimiento adecuado de las técnicas más importantes y de su potencial. Ello facilitará su relación con los biotecnólogos, y su integración en equipos de investigación multidisciplinarios. La política del INIA y de CORPOICA en este sentido ha sido la de establecer este tipo de equipos, como una forma de promover y facilitar la interacción entre las distintas personas requeridas para llevar adelante un programa de biotecnología con orientación productiva. Planificación y priorización Este problema es especialmente importante para los institutos de investigación agrícola, debido a que, por lo general, no cuentan con biólogos. Para desarrollar capacidades en biotecnología pueden especializar alguno de sus ingenieros agrónomos o médicos La IyD en biotecnología, es casi por definición orientada hacia la solución de problemas prácticos, muchos de los cuales son del ámbito productivo. La selección de las líneas y proyectos de investigación debería realizarse, en consecuencia, mediante una planificación o 32 priorización que tome en cuenta el mercado o la opinión de productores o industriales. El carácter de servicios que tiene mucha de la investigación que se puede realizar en biotecnología obvia el problema de la priorización, que se traslada al de las líneas o proyectos de investigación a los que se presta servicios. interrelacionadas entre distintas instituciones. Ello es producto de la actividad planificadora y coordinadora del organismo nacional de promoción de ciencia y tecnología, COLCIENCIAS, a través de su Programa Nacional de Biotecnología, que ha sido exitoso en orientar y coordinar muchos de los grupos de investigación existentes en el país. Este es el caso de los dos institutos de El Salvador analizados, que no tienen un problema de priorización pues sus unidades de biotecnología prestan servicios a otras unidades o a clientes externos. En el INIA, los proyectos en biotecnología son solicitados o concertados con los programas, que a su vez responden a lineamentos de las instancias directivas de la institución, en las que los productores están fuertemente representados. La Unidad también presta servicios técnicos y de investigación contratada a clientes directamente. Vinculación con la industriaVinculaci—n con la industria Con excepción del CENTA, todos los demás institutos estudiados prestan servicios directamente a la industria, sean éstos de asistencia técnica o de investigación contratada. Sin embargo, las dificultades administrativas de esta actividad dentro de las organizaciones está llevando a la búsqueda de formas más adecuadas para esta relación con el sector productivo. En el INIA existe un proyecto de creación de una empresa especializada en venta de servicios de biotecnología, denominada INIA-BIOTEC. La Universidad Nacional de Colombia, por otro lado, creó recientemente una empresa para realizar investigación por contrato y prestar servicios denominada Corporación Industrial de Biotecnología, en conjunto con CORPOICA, el Instituto Nacional de Salud y 15 empresas y organizaciones del sector privado. También en CORPOICA, la selección de proyectos responde a lineamentos generales concertados con los productores. Los programas de biotecnología pueden realizar investigación contratada o concertada con otros organismos. Esta última opción es importante en Colombia, donde un análisis de la programación de investigación de varios institutos evidencia la existencia de actividades 33 Necesidades de apoyo al desarrollo de la biotecnología agrícola: Conclusiones y recomendaciones La introducción y consolidación de la agrobiotecnología en los países de la región es un proceso complejo y costoso que exige un conjunto de acciones coherentes en el tiempo especialmente dirigidas a tal fin. La experiencia de la región en esta materia evidencia una serie de debilidades que sugieren, a su vez, cambios en las estrategias y nuevas formas de enfrentar los problemas. Los países que ya cuentan con capacidades en biotecnología enfrentan el reto de consolidar y orientar los esfuerzos hechos en función de objetivos industriales y comerciales, sociales y ambientales específicos. La mayoría de ellos no han podido aún superar la fase “científica” del desarrollo de la biotecnología. Las capacidades y esfuerzos aún se encuentran principalmente en el sector académico y de investigación, sin relaciones con el sector productivo o de servicios. Se exceptúan algunas técnicas, como el cultivo de tejidos, por ejemplo, bien difundidas y utilizadas en algunas industrias. Un grupo importante de países de la región aún no cuentan con las bases científicas y profesionales mínimas para enfrentar los retos que plantea la biotecnología. Todavía existe, por lo tanto, un amplio campo de posibilidades para programas de creación de capacidades científicas de primera generación, es decir, con un fuerte énfasis en la formación de recursos humanos y la creación de una infraestructura técnica y científica mínima.	30,888
W2942926940.txt_1	German-Science-Pile	Open Science	Various open science	null	None	None	German	Spoken	1,343	2,134	K l c i n e Mitteilung'en. 224 Q u e r s e h n i t t des W u l s t e s ist in seinem u n t e r e n Tell aleutlicit siehtbar u n d bildet eine Spirale yon m e h r e r e n Windungem Seine Aehse i s t eine offene Kurve, in deren Sinne die in Fig. l a n n d l b gezeiehneten hellen u n d dunklen Streifen verlaufen. Diese begleiten die Spiralflamme u n d bilden m i t dem Q u e r s c h n i t t des W u l s t e s n u t eine E r s e h e i m m g . Streifen a n d Spirale bedingen sich gegenseitig. Die Bewegung des Gasstromes in der Spirale erfolgt yon autaen naeh innen; dabei geht die ]/ l~ig. 1 a. lir~ie Su auf, deren Windunge~i a a t e r giinstige~ Umstrtnden einen Durct~messer yon m e h r e r e n Millimeterll erreichen. Die Schraubenlinie aber zerfii.llt in eine Reihe leuchtender ]?unkte, die in der B a l m der urspriingtichen Schranbenlinie sich welter bewegen bis zu dem Augenblick, wo sie durch vollstiindige V e r b r e n n u n g versehwinden (Fig, 2c u n d 2d). Die Bildung fester Kohlenstoffkiigelchen ist auch sonst schon bet Gasexplosionen beobachtet worden, z. ]3. bet der Explosion in der Radbodgrube zu Westfalen a m 12. November 1908. Der bet dieser E r s c h e i n u n g anftretende Spiralwirbel zeigt grotae Ahnlichkeit m i t den photographischen Bil dern, die wir einerseits yon Spiralnebeln, andererseits yon einigen Kometen besitzen, u n d mail k a n n hiernach vermuten, da,$ diese kosmischen Gebilde ihr Aussehen aus dem A u f e i n a n d e r t r e f f e n yon zwei sich m i t versehiedenen Geschwindigkeiten fortbewegenden Gasma,ssen erhalten haben. (Comptes Rendus 154, 698, 1156 und 155, t243, 1912.) MIc. Fig. 1 b. Spirale yon ether leuehtenden Fl~iehe aus, die sieh allm~thlich verengt. W e n n der G a s s t r a h l n u t den hellleuehtenden Tell der Flamme trifft, bildet sich der W u l s t ohne Spirale u n d ohne Streifen. Diese entstehen e r s t beim A u f t r e f f e n des Strahles auf den danklen Tell der Flamme. W i r d sein Druck so erhSht, data er bis znr Basis der F l a m m e reicht, so vergrSl3ert sich die Zahl der W i n d u n g e n in der Spir~le bis auf 5 oder 6. Ebenso verm e h r t sich die Zahl der Streifen. Diese E r s e h e i n u n g erkl:,trt sich so~ data (lurch den Slob des Gasstrahles die den dunklen Tell der Fl~mme umgebende bl~tulich leuchtende Schicht u m sich selbst m i t dem dunklen I n n e r n der Flamme z u s a m m e n aufgerollt wird, wie der K a m m einer brandenden Welle: Der leuchtende W u l s t ist gerade das Autaere dieser Gaswelle. E r bildet also eine aus zwei gesonderten Sehichten bestehende Rolle. Seinen Q u e r s c h n i t t zeigt die leuchtende Spirale, in der Vorderal~sicht dagegen zeigt er die Streifen, von denen jeder einer W i n d u n g der Spirale entsprieht. W i r d der Druek Fig. 2 c Fig. 2 d. Fig. 2a. Die Nat m'[ ~ iss(msehafttul Fig. 2b. der Flamme so gesteigert, data er 1/10 yon dem des Gasstrahles betr~gt, so ';indert sich die Erscheinung. U n t e r dem W i d e r s t a n d e der F l a m m e biegt sich die Spitze des G~sstrahles G (Fig. 2 a u n d 2b) zuriick. Das leuchtende Z e n t r u m der Spirale ~p vergriifiert sich zu ether runden Scheibe; es g e h t yon gelber zll rSttieher F~trbung tiber, u n d dieser innere rote Tell dehnt sieh I~ngs der Achse des W u l s t e s W aus. E r l~ist sich in eine Schr~ubea- Der Sommer 1912 h a t den Bewobnern yon Mitteleuropa zwei i n t e r e s s a n t e meteorologische Ph~tnomene gebracht: den betrliblichen R e i e h t u m an Niederschl'Xgen sowie die merkwiirdige Triibung der Atmosph~ire, die auch an ganz wolkealosen Tagen den Himmel nieht blau, sondern well]Itch erscheinen lieB. W ~ h r e n d die MeteoroIogie fiber die e r s t g e n a n n t e E r s e h e i n u n g sich vorerst n u t in V e r m u t u n g e n ergehen kann, ist das zweile Phitnomen m~ch der z u s a m m e n f a s s e n d e n D'~rsteltung des I I e r r a HeIlmann vollkommen gekl~irt. (Die Ursache der ungew6hnlichen Tri~bung der Atmosphiire im Sommer 1912. Met. Zeitschrift 1913 S. 34.) Die Anom~lie im Aussehen des IIimmels, die insbesondere aueh in den ungewShnlich roten Sonaenauf- a n d Unterg~ngen zum A u s d r u e k kam, riihrte her yon einem a m 6. bis 8. J u n i erfolgten A u s b r u e h des bisher als erloschen angesehenen Vulkans K a t m a i auf der Halbi~lsel Alaska. Dabei wurden gewaltige A s e h e n m e n g e a i n die L u f t geworfen. A u f dem Mount-Weather-Observatorium (bet W a s h i n g t o n ) wurde die Trfibung am 10. J u n i bemerkt. Die in den gemi~Bigten Breiten vorherrsehenden Westwinde t r u g e n die Staubteilchen fiber den Ozean nach Europa. A m 21. J u n i beobavhtete de Quervaln auf dem Inlandeis yon GrSnland den merkwiirdigen Anbliek des Himmels, auf dera K o n t i n e n t e wurde die Triibung in der Zeit vom 23. bis 27. J u a i zum ersten Male k o n s t a t i e r t. Das Ph~nomen e r i n n e r t an den gewaltigen Ausbruet~ des K r a k a t a u im J a h r e 1883, wobei die Staubmassen mehrere Male die Erde u m w a n d e r t e n . Es ist daher nicht ausgeschlossen, dal] aud~ im vorliegenden Falle die inzwischen abgewanderten Trfibungen nochmals wiederkehren. I n den Kreisen der P h y s i k e r wurde die Frage viel diskutiert, ob nicht zwisehen dem Regenreichtum des Soremers 1912 u n d den Trtibungen der Atmosph~ire ein Zus a m m e n h a n g besteht. Man e r i n n e r t sieh der Tatsaehe, dab zur Wolken- u n d Niedersehlagsbildung sogenannte K o n d e n s a t i o n s k e r n e notwendig sind, d. h. FremdkSrper, an welchen der W a s s e r d a m p f der Atmosph[tre sich anlagern kann. 0 h n e zu der Frage Stellung nehmen zu wollen, weist H e r r HeIlmann auf das J a h r 1783 hin, das trotz ether ~i.hnlichen, yon einem Y u t k a n a u s b r u c h auf Island herriihrenden Trfibung tier Atmosph~i~re einen uugewShnlieh heiBen Sommer halle. A. Schmauss, Miinchen. Fiir die Redaktion verantwortlich: Dr, Arnold Berliner. Berlin W.9,.	3,237
https://openalex.org/W3112302437_1	Spanish-Science-Pile	Open Science	Various open science	2,020	La toma de decisiones de gobiernos y sectores de la sociedad, intuiciones y datos en tiempos de pandemia SARS-COVID-19	None	Unknown	Unknown	1,116	7,945	Alida María Acosta Ortiz Revista de la Facultad de Ciencias de la Salud, Universidad Autónoma de Bucaramanga (Colombia) Vol. 23 Número 3 - Diciembre 2020 - Marzo 2021 ƠƓƤƗơƢƏƒƓƚƏƔƏƑƣƚƢƏƒ ƒƓƑƗƓƜƑƗƏơƒƓƚƏơƏƚƣƒ i-ISSN 0123-7047 - e-ISSN 2382-4603 https://doi.org/10.29375/issn.0123-7047 Infecciones urinarias Agotamiento profesional Obra titulada: “Le docteur Paul Gachet [El doctor Paul Gachet]”, 1890, por: Vincent van Gogh (1853-1890) Depresión Editorial Vol. 23(3): 369-371, diciembre 2020 - marzo 2021 '2,KWWSVGRLRUJ La toma de decisiones de gobiernos y sectores de la sociedad, intuiciones y datos en tiempos de pandemia SARS-COVID-19 Decision-making by Governments and Sectors of Society, Intuitions and Data in Times of the SARS-COVID-19 Pandemic A 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https://openalex.org/W2884480969	OpenAlex	Open Science	CC-By	2,018	Affect and ipsative approaches as a counter to pedagogic frailty: the guardian of traditional models of student success	Julie Rattray	English	Spoken	8,320	13,439	Dr. Julie Rattray School of Education. University of Durham School of Education. University of Durham University of Durham, School of Education. Leazes Road, Durham. DH1 1TA. UK Word count: 6585 julie.rattray@durham.ac.uk Affect and ipsative approaches as a counter to Pedagogic Frailty: The guardian of traditional models of student success Abstract In this paper I consider how the neoliberal discourses surrounding Higher Education have resulted in an increasingly risk averse culture of learning and teaching. Students are frequently reluctant to engage with troublesome or challenging knowledge and academics are less likely to push learners into contested spaces or deviate from excepted pedagogical practices for fear of upsetting them. The consequences of this situation are that we potentially have a generation of graduates who lack the resilience to cope in the graduate market. Drawing on the notion of pedagogic frailty consideration of how models of success that are associated with high stakes, single point assessment, might limit the development of positive affect in learners. I argue instead for a reconsideration of notions of success building on the principles of ipsative assessment as a means of supporting the development of affective attributes, such as resilience, optimism and hope, in an effort to ensure graduates are equipped for an uncertain future. Keywords: Assessment, pedagogy, emotions, student learning, teaching Introduction Introduction Changes in the Higher Education landscape over the past two decades have resulted in an increasing identification of students as consumers and universities as service providers (Barnett, 2011; Klemenčič, 2011; Molesworth, Scullion & Nixon, 2009; Scullion, Molesworth, & Nixon, 2011; Williams, 2013; Woodall, Hiller, & Resnick, 2014). Higher Education, now part of a market economy (Barnett, 2011), has become over-shadowed by threats of litigation and value for money, resulting in the emergence of new and hitherto unforeseen challenges to both students and academics as they strive to achieve and foster success. (Ball, 2003; Barnett, 2011; Kaye, Bickel & Birtwistle, 2009; Naidoo & Jamieson, 2005; Stensaker & Harvey, 2011). The idea that Higher Education is seen as a place of challenge for learners where they are confronted by troublesome knowledge (Perkins, 2006; 2008) designed to stretch their thinking and facilitate their understanding is no longer universally acceptable. (Delucchi & Korgen 2002; Land, 2017; Williams, 2013; Instead we are confronted by students who ‘expect’ the success they are paying for (Delucchi & Korgen, 2002; Regan, 2012; Shepperd, 1997), and an academic body that is increasingly reluctant to push learners into uncomfortable and troublesome places for fear of ‘upsetting’ them (Kinchin, 2015). The resultant risk averse culture that is emerging seems to be at odds with the demands of employers who want creative, flexible, critical thinking graduates who are responsive to the demands of an ever-changing professional landscape (Barrie, 2006; Butterwick & Benjamin, 2006). In short, employers need graduates who have developed a set of graduate competencies or attributes that equip them with the intellectual skills and dispositions needed for a rapidly changing and uncertain future (Bowden, Hart, King, Trigwell & Watts, 2000). In addition, they want graduates who have developed the requisite psychological or affective attributes that ensure they can deal with the Changes in the Higher Education landscape over the past two decades have resulted in an increasing identification of students as consumers and universities as service providers (Barnett, 2011; Klemenčič, 2011; Molesworth, Scullion & Nixon, 2009; Scullion, Molesworth, & Nixon, 2011; Williams, 2013; Woodall, Hiller, & Resnick, 2014). Higher Education, now part of a market economy (Barnett, 2011), has become over-shadowed by threats of litigation and value for money, resulting in the emergence of new and hitherto unforeseen challenges to both students and academics as they strive to achieve and foster success. Introduction (Ball, 2003; Barnett, 2011; Kaye, Bickel & Birtwistle, 2009; Naidoo & Jamieson, 2005; Stensaker & Harvey, 2011). The idea that Higher Education is seen as a place of challenge for learners where they are confronted by troublesome knowledge (Perkins, 2006; 2008) designed to stretch their thinking and facilitate their understanding is no longer universally acceptable. (Delucchi & Korgen 2002; Land, 2017; Williams, 2013; Instead we are confronted by students who ‘expect’ the success they are paying for (Delucchi & Korgen, 2002; Regan, 2012; Shepperd, 1997), and an academic body that is increasingly reluctant to push learners into uncomfortable and troublesome places for fear of ‘upsetting’ them (Kinchin, 2015). The resultant risk averse culture that is emerging seems to be at odds with the demands of employers who want creative, flexible, critical thinking graduates who are responsive to the demands of an ever-changing professional landscape (Barrie, 2006; Butterwick & Benjamin, 2006). In short, employers need graduates who have developed a set of graduate competencies or attributes that equip them with the intellectual skills and dispositions needed for a rapidly changing and uncertain future (Bowden, Hart, King, Trigwell & Watts, 2000). In addition, they want graduates who have developed the requisite psychological or affective attributes that ensure they can deal with the associated uncertainty, complexity and challenge. It should be acknowledged here that such claims pose a direct challenge to the dominant neoliberal discourse pervading Higher Education, particularly in the UK and USA, which is predicated on the idea that Higher Education provides a sound preparation for the world of work (Naidoo & Williams, 2015). In challenging these assertions we need to consider how students and academics are positioned within the neoliberal university and the extent to which the increasingly restrictive learning environment actually limits rather than promotes readiness for work. This paper explores some of these tensions in relation to student success, focusing on how students and academics might be encouraged to stem the tendency towards risk-averse behaviours by developing affective attributes, such as resilience and hope that will support academic risk taking and confrontation with troublesome knowledge and liminal spaces. In this paper I argue that in thinking about successful learning rather than success, students might be supported to develop complex learning skills and affective competencies that equip them for their uncertain futures beyond the academy. Introduction By moving away from the individualised, outcome focused notion of success as predicated by a neoliberal approach to education (Ingleby, 2013; Saunders, 2010), I argue for success to be considered in relation to academic practices and approaches to pedagogy which have the potential to limit or foster success. Drawing on notions of pedagogic frailty and ipsative assessment I explore the extent to which the constraining influences of the neoliberal discourse in Higher Education might be redressed. Of course, learning is influenced by more than academic practices and pedagogies i.e. issues of student finance and debt, mental health and career aspirations of learners will all influence how students engage with learning and impact upon notions of success. Of principal concern here is the move towards increasingly ‘safe’ pedagogies and how they might be avoided. I am not suggesting that other factors are unimportant or irrelevant but rather offering a consideration of one view of success as a counter to the seemingly dominant, and arguably oppressive, neoliberal discourse within Higher Education (Clegg, Hudson & Steel, 2010; Morley & Dunstan, 2013). avoided. I am not suggesting that other factors are unimportant or irrelevant but rather offering a consideration of one view of success as a counter to the seemingly dominant, and arguably oppressive, neoliberal discourse within Higher Education (Clegg, Hudson & Steel, 2010; Morley & Dunstan, 2013). Pedagogic Frailty: the guardian of traditional models of success Kinchin and Winstone (2017) argue that this risk averse culture in Higher Education is symptomatic of what they term ‘pedagogic frailty’ (Kinchin, 2015; 2016a; 2016b), with learners and academics preferring ‘safe’ and uncontested spaces. Academics, they argue, no longer push students into troublesome encounters, opting instead to stick to tried and tested content and pedagogies which may require work on the part of the student but which keep them within largely familiar and comfortable learning spaces (Land, 2017). In so doing, success in the traditional sense may be more readily attainable but students are less likely to develop the intellectual and affective competencies needed for success beyond the academy (Darling-Hammond & Snyder, 2000). In talking about traditional models of success in this paper I am referring to models which emphasise the grades students obtain and take these as a single measure of competence (Astin & Antonio, 2012; Biggs, 2003; Elton & Johnston, 2002; Gibbs, 2006; Hussey & Smith, 2003). Affective attributes like resilience and hope are not likely to be fostered in students who encounter only limited challenges in their learning or who never stray outside of what is safe (Rattray, 2016) Students who do not experience learning situations which expose them to the complexities of their disciplines are less likely to be able to cope with complexity later. We must ask therefore if such practices are really fostering success in a meaningful way. As we shall see later in the paper such practices are not reflective of academics’ inability to provide challenging learning experiences. They are a consequence of the pressures exerted on academics by the neoliberal doctrine of accountability, consumerism and marketisation in an academy governed by league tables (Barnett, 2011; Findlow, 2006; Howard, 2013; Naidoo & Williams, 2015; Williams, 2013). According to Kinchin (2017) pedagogic frailty offers an explanation for the predominance of traditional approaches to teaching, learning and assessment. Lectures, seminars, exams and essays continue to make up the dominant pedagogy for many students, despite these being at odds with the beliefs held by academics in relation to how people learn and what constitutes a positive learning experience (Howard, 2013). To this end it provides an explanation as to why success continues to be associated with graded performance on assessments, typically designed to measure the outcomes of learning through a set of predetermined criteria relating to knowledge that is quantified (Biggs, 2003 Jackson, 2000). Pedagogic Frailty: the guardian of traditional models of success The reluctance of academics to rethink success in terms of not just the end products but to focus on the learning process itself and the associated academic and psychological attributes as an indicator of success is a consequence of frailty (Winston, 2017). Kinchin (2017) argues that such ‘pedagogic frailty results from the degraded quality and reduced extent of interactions within and between aspects of the professional environment’ (p. 11). In particular four key aspects of the professional academic environment (see below) are associated with the emergence of pedagogic frailty (Kinchin, 2015). In all cases it is the extent to which there is a convergence of beliefs or disharmony which seems to result in frailty. It is important to note that frailty is not a binary or unitary state, as a systemic condition it varies as the connections between individuals and departments within the system fluctuate. It is dependent on the levels of harmony and variance along the four dimensions identified by Kinchin (2015) as contributing to frailty. None of the dimensions alone evoke frailty but will interact with each other, the discipline and the institutional environment to create a sense of frailty or otherwise. The first two dimensions associated with pedagogic frailty identified by Kinchin (2015) relate to the convergence of individual and/or disciplinary ideas about teaching and learning with that of the institution. The extent to which an individual’s beliefs and values in relation to teaching and learning correspond to the approaches to teaching, learning and assessment they may be required to adopt within their institutional context is the first of these. Such values are discussed infrequently and as a consequence the potentially shared, or not, nature of them is rarely actualised (Mårtensson & Roxå, 2016). Kinchin (2017) acknowledges that the use of the term ‘frailty’ is not uncontentious evoking in us, as it does, a sense of weakness or vulnerability. His choice of term is however, no accident, using the term, as he does, to provoke in us an emotional response which renders us uncomfortable. He argues that developing the requisite resilience that is needed to take risks in our teaching is not easy and frequently takes us into uncomfortable places. Confronting our discomfort and engaging in discussions designed to support risk taking and resilience is an important part of combating the frailty (Mårtensson & Roxå, 2016). Pedagogic Frailty: the guardian of traditional models of success A bringing to the fore of such shared values could, according to Barnes (2014), create a greater sense of resilience within the academy. It is not just individual values however that might be at odds with the dominant pedagogies of an institution. The extent to which disciplinary practices are mirrored by institutional pedagogy is also relevant here in determining frailty and is Kinchin’s second dimension. Of particular relevance here is the idea of authenticity and the extent to which the learning opportunities being offered to students are perceived to provide authentic learning experiences by academics (Kinchin, 2015; Kreber, 2010; Pawlina & Drake, 2016). If academics do not believe they are able to offer authentic learning experiences that will equip students for success in their chosen careers, in part, because of accepted institutional or even traditional disciplinary pedagogies which might be outdated and beyond their power to change, they will be less likely to try. The third dimension of importance in the pedagogic frailty model is that of the research-teaching nexus (Kinchin, 2015). Whilst the model is not predicated on a particular view of this nexus it posits that the extent to which the individual academic’s view of the research-teaching relationship is supported or at odds with institutional culture will serve to promote or discourage frailty (Kinchin, 2015). The final dimension of relevance to the model is that of locus of control (Kinchin, 2015). The increasing marketisation of Higher Education has resulted in a new form of managerialism in Higher Education which has seen decisions becoming increasingly centralised with less autonomy being given to the individual academic (Jones, 2014). This has resulted in decreasing levels of autonomy for academics promoting a reluctance to take risks as levels of accountability increase (Olsen, 2016). Creative pedagogies become risky and resilience is degraded as academics experience an increasingly top-down approach to the design, delivery and assessment of the courses they teach, in short increasing levels of managerialism (Findlow, 2008) constrain change and encourage a culture of ‘playing it safe’. Pedagogic frailty has largely been explored from the perspective of the academic providing explanations for risk-averse behaviour and declining resilience in relation to teaching and learning (Kinchin, 2017; Winstone, 2017). If teaching becomes increasingly ‘safe’, with academics less likely to try new approaches to delivery or to think about different models of assessment or ways to conceptualise success (Spooren. Pedagogic Frailty: the guardian of traditional models of success Brockx, & Mortelmans, 2013), learners will experience learning where they know the ‘rules’ and can follow them, where they themselves no longer take risks, where they perform a specified task focusing on the end result not thinking about the process of how the end result is attained or other ways to do this. In short, they become pedagogically frail as they stay within the limits, never learning to cope with set-backs in their learning or adapt to challenging situations. All they do is attainable and the rules are clear. To this end success is measured in test scores and final grades based on how much they know and how well they can solve recognisable problems. Such a view of success may ensure a good degree outcome but it is potentially limiting outside of the academy where the rules may be less clear and problems do not present themselves in predictable and familiar ways. To be successful in the graduate market future graduates may be confronted with problems that are as yet unknown. When faced with previously unseen problems success will be dependent not on the ability to follow a well-learned and rehearsed formula but rather on the ability to adapt and be resilient in the face of challenge. ‘rules’ and can follow them, where they themselves no longer take risks, where they perform a specified task focusing on the end result not thinking about the process of how the end result is attained or other ways to do this. In short, they become pedagogically frail as they stay within the limits, never learning to cope with set-backs in their learning or adapt to challenging situations. All they do is attainable and the rules are clear. To this end success is measured in test scores and final grades based on how much they know and how well they can solve recognisable problems. Such a view of success may ensure a good degree outcome but it is potentially limiting outside of the academy where the rules may be less clear and problems do not present themselves in predictable and familiar ways. To be successful in the graduate market future graduates may be confronted with problems that are as yet unknown. Pedagogic Frailty: the guardian of traditional models of success It emphasises what is learned not how learning happens and can often be associated with prescriptive approaches to learning and pedagogy (Black & Wiliam 1998; 2003). Such approaches, I argue, do not support the development of complex academic and affective capabilities. Pedagogic Frailty: the guardian of traditional models of success When faced with previously unseen problems success will be dependent not on the ability to follow a well-learned and rehearsed formula but rather on the ability to adapt and be resilient in the face of challenge. ‘rules’ and can follow them, where they themselves no longer take risks, where they perform a specified task focusing on the end result not thinking about the process of how the end result is attained or other ways to do this. In short, they become It would seem to be time to reconsider normative views of success linked as they are to a set of fixed learning outcomes and judgements, and questions about whether these are achieved or not (Biggs, 2003: Regan, 2012; Sadler, 2005; Stobart, 2008). It would seem to be time to reconsider normative views of success linked as they are to a set of fixed learning outcomes and judgements, and questions about whether these are achieved or not (Biggs, 2003: Regan, 2012; Sadler, 2005; Stobart, 2008). Typically normative views of success are associated with single, high stakes, assessment points that come at the end of a module or course and do not try to determine individual progression as students work through the curriculum content (Astin, 2012; Bloxham & Boyd, 2009). The consequences of normative models of success are that they frequently encourage students to search for model or ‘accepted’ answers to questions and to adopt ‘safe’ interpretations of ideas, rather than offering more individual or creative responses (Furedi, 2011). Such approaches may ensure good grades but they do not encourage students to take risks in their learning for fear of obtaining a poor grade (Price & O’Donovan, 2006; Price, Handley, Millar & O'Donovan, 2010; Price, Carroll, O'Donovan & Rust, 2011). Success becomes linked to grades and performance and is associated with the outcomes of learning rather than the process of learning. It emphasises what is learned not how learning happens and can often be associated with prescriptive approaches to learning and pedagogy (Black & Wiliam 1998; 2003). Such approaches, I argue, do not support the development of complex academic and affective capabilities. obtaining a poor grade (Price & O’Donovan, 2006; Price, Handley, Millar & O'Donovan, 2010; Price, Carroll, O'Donovan & Rust, 2011). Success becomes linked to grades and performance and is associated with the outcomes of learning rather than the process of learning. Ipsative Approaches and the development of affect In the introduction to this paper I argued that it is important for learners to engage with troublesome knowledge (Perkins, 1999; 2008). If learners are to develop resilience they need to experience that which is challenging, ideas which will stretch them, concepts which are not immediately available to them. In short they need to learn not simply to understand the material presented to them but how to understand it and what to do when this does not happen immediately or easily. Only through challenging encounters can learners develop resilience, the ability to minimise the long-term emotional consequences of negative learning experiences (Rutter, 2006). Resilient learners are able to cope with setbacks to their learning and move on. They do not dismiss negative learning experiences as unimportant or inconsequential but rather, they are able to evaluate the situation and reflect on why it occurred. They do not internalise the negative experience as a personal measure of worth (Martin & Marsh, 2009; Nota, Soresi & Zimmerman, 2004). Such learners are able to accept the part they themselves may have played in the negative learning experience and identify ways to adjust future learning behaviours to minimise the potential repetition of the experience (Nota, Soresi & Zimmerman, 2004). Resilience is not only developed through encounters with troublesome knowledge it will also support future encounters with such knowledge Soresi & Zimmerman, 2004). Such learners are able to accept the part they themselves may have played in the negative learning experience and identify ways to adjust future learning behaviours to minimise the potential repetition of the experience (Nota, Soresi & Zimmerman, 2004). Resilience is not only developed through encounters with troublesome knowledge it will also support future encounters with such knowledge (Rattray, 2016). Resilient learners who have experienced difficulties mastering challenging materials in the past but who have overcome such difficulties will be more willing to engage with future encounters that involve troublesome knowledge. An ability to deal with negative learning experiences is not the only affective competence that is important to the learning process. Work in the area of positive psychology suggests that other affective attributes also serve to facilitate learning (Lopez and Snyder, 2003; Seligman, 2006; Seligman, Ernst, Gillham, Reivich and Linkins, 2009). Research in this area highlights the importance of academic hope and optimism as important affective attributes that will facilitate and further learning. Ipsative Approaches and the development of affect Academic hope serves as a mediator of problem solving abilities (Chang, 1998) and a predictor of future academic success that is more powerful than intelligence (Davidson, Feldman & Margalit, 2012; Day, Hanson, Maltby, Proctor & Wood, 2010). Academic hope refers to an individual’s acceptance of their own agency in learning and their ability to identify and follow multiple pathways to the attainment of a distal goal (Snyder, Shorey, Cheavens, Pulvers, Adams III & Wiklund, 2002). Optimism relates to academic hope, emphasising as it does the importance of personal agency and self- determination as a means of achieving positive outcomes (Bryant & Cvengros, 2004; Shogren, Lopez, Wehmeyer, Little & Pressgrove, 2006). Taken together, resilience, optimism and academic hope, with their emphasis on personal agency, identification of multiple pathways to attain goals, a belief in positive future success and the ability to minimise the negative consequences of setbacks, offer an appealing skill set to future employers who want graduates prepared to deal with the unknown challenges they may encounter. Resilience, optimism and academic hope are not static attributes, rather they are developed over time in response to learning experiences (Snyder, Shorey, Cheavens, Pulvers, Adams III & Wiklund, 2002). As we accumulate learning experiences they build what Cousin (2006) has called ‘emotional capital’. The importance of emotional capital for learning is that it serves as a source of support, motivation and even sustenance to enrich and sustain learning when difficulties and struggles are encountered. Learners who are high in emotional capital will persist in the phase of challenge and adversity and alter their approach in the face of an unsuccessful first attempt. They understand that mastery of a learning goal may not be achieved at the first attempt and can identify alternatives for a second or even third attempt. Learners who lack emotional capital, perhaps the learners’ equivalent of pedagogic frailty, lack the ability to identify alternatives, are unable or unwilling to persist in the face of challenge and frequently give up before the learning task is achieved or concept mastered. Success in conventional terms for these learners is frequently out of reach and continued encounters with failure, or even perceived failure, in the shape of low grades simply serves to denigrate the emotional capital further. Re-framing success In this paper I argue for a reframing of success in ipsative terms as a means of supporting the development not only of students’ academic skills but also their affective attributes and in so doing encouraging students, and indeed academics, to become less risk-averse. Ipsative approaches to assessment and learning offer an alternative conceptualisation of the learning process and how it is experienced by both learners and academics. Whilst research in the area is still relatively limited I contend that it has much to offer to our discussions of success and could serve as a counter to pedagogically frail practices. It offers academics a greater opportunity to reflect on the ongoing learning of students and to have a sense of where the student is in the learning process. Such opportunities opening up, as they do, a more elaborate discourse between learner and academic (Hughes 2011; 2014) might encourage academics and learners to become less risk-averse as they experience the learning together and have a greater sense of what is to be learned and how it might be learned. As academics come to see that learners can cope with challenge they may introduce greater challenge into their teaching (Howard, 2013; Le Fevre, 2014), becoming less risk averse. Likewise learners who cope with minor setbacks become less risk-averse too. Thus ipsative approaches have the potential to counter the risk-averse climate as academics and learners engage in learning experiences that are bounded by trust (Cozilino, 2014). Ipsative models of success offer a move away from normative or criterion referenced assessment at the end of a programme of study and focus instead on the ongoing learning experiences and accomplishments of the individual learner as they progress through the programme of study (Hughes, 2011; 2014; Hughes, Wood & Kitagawa, 2014; Hughes, 2017). The ipsative approaches to success utilise an approach to assessment that encourages students and academics to reconceptualise success as incremental and cumulative, something that is built gradually over time rather than something that is a single fixed measure of performance on a given day. Ipsative approaches encourage learners to reflect on their own progress and accomplishments, thinking about the process of learning as much as the product. Re-framing success In so doing learners are required to engage in ongoing self-reflection facilitated by discursive and dialogic feedback experiences that allow them to think about where they are in relation to their own understanding of what is being learned and where they need to go (Hughes, 2014). Success in such models may still include a final grade which is linked to the learning outcomes of a module but it is achieved through an iterative and recursive learning experience which has the potential to support risk taking as students see successful learning as something that is ongoing and can be associated with some wrong turns and difficult encounters along the way. Students who have the opportunities to make mistakes and mis-steps are able to build resilience and emotional capital as they learn to cope with these ‘failures’ and move on. Ipsative models of assessment and success create safe spaces for learners to explore learning. They are not safe in the sense that they offer no challenge but rather they are safe in that they provide the space to explore an idea and make mistakes. They remove the necessity that the first attempts to demonstrate mastery of the concept is the one that will be graded and serve as an indicator of success (Hughes, 2011; 2014; 2017). Many traditional approaches to assessment take this latter approach offering at most one opportunity for formative feedback on a small piece of work before the final summative assessment is graded (Bloxham & Boyd, 2009). Such approaches are high stakes and learners frequently seek sample or model answers or accepted ways of approaching the assessment for fear of getting a low grade. Ipsative approaches offer the opportunity for multiple feedback experiences, and perhaps more importantly, support students in their abilities to self-assess and think about what is going on in their learning (Hughes, 2014). Ipsative models of assessment and success create safe spaces for learners to explore learning. They are not safe in the sense that they offer no challenge but rather they are safe in that they provide the space to explore an idea and make mistakes. They remove the necessity that the first attempts to demonstrate mastery of the concept is the one that will be graded and serve as an indicator of success (Hughes, 2011; 2014; 2017). Re-framing success Many traditional approaches to assessment take this latter approach offering at most one opportunity for formative feedback on a small piece of work before the final summative assessment is graded (Bloxham & Boyd, 2009). Such approaches are high stakes and learners frequently seek sample or model answers or accepted ways of approaching the assessment for fear of getting a low grade. Ipsative approaches offer the opportunity for multiple feedback experiences, and perhaps more importantly, support students in their abilities to self-assess and think about what is going on in their learning (Hughes, 2014). Rather than simply providing students with a grade and information about what might be wrong with the work they encourage students to engage with their own work and assess where they are in their own learning. In so doing students come to understand how to evaluate where they are in terms of their learning and where they need to go. In short, it helps students to establish learning goals and through their attainment potential paths to achieve future goals. In so doing the emotional capital (Cousin, 2006) of the learner is developed as they build confidence in their abilities to achieve goals through multiple pathways and their own agency. They come to see and understand who they Rather than simply providing students with a grade and information about what might be wrong with the work they encourage students to engage with their own work and assess where they are in their own learning. In so doing students come to understand how to evaluate where they are in terms of their learning and where they need to go. In short, it helps students to establish learning goals and through their attainment potential paths to achieve future goals. In so doing the emotional capital (Cousin, 2006) of the learner is developed as they build confidence in their abilities to achieve goals through multiple pathways and their own agency. They come to see and understand who they are as learners, which facilitates their ongoing planning and assessment of future learning. Ipsative approaches facilitate engagement with troublesome knowledge as learners build the affective and academic competencies that will support their engagement with complexity and encourage them to accept liminality as an important part of the learning experience (Rattray, 2016; 2017). Re-framing success In offering the case for ipsative approaches I acknowledge Hughes’s (2011; 2014) assertion that it will be initially effortful and as such may be at odds with neoliberal managerial proclivities which emphasise pedagogical approaches that are uniform and fit easily within the audit and accountability culture (Barnett, 2011; Williams, 2013). It is possible however, to start making small inroads in to the approach utilising formative assessment opportunities as a first step (Hughes, 2011). We can start by asking students to add a reflective comment to any formative, and indeed summative, work they submit. This reflective comment should offer a qualitative remark on the students’ experience of actually producing the work being reviewed; how did they prepare? what did they find difficult? what are they still unsure about? what do they feel they have done well?. Such a comment opens up a potential dialogue with the marker who, in the course of providing normal written feedback can offer a direct response to the reflective comments. To ensure the approach is truly ipsative before grades are released to students they may be asked to respond to the assessors’ feedback once more, this time offering a reflective response to the feedback itself and what it tells them about where they are on their learning journey. If we build smaller more cumulative assessment opportunities in to our teaching then this process can gradually be developed so that it becomes an increasingly meaningful dialogue. In addition once students become familiar with the process it can be extended to include peer review and feedback as an alternative source of comment and reflection. Conclusion In this paper I have argued current climates in Higher Education have resulted in what Kinchin (2015) calls pedagogic frailty. Academics are becoming increasingly risk- averse against a backdrop of increased marketisation and accountability. Fears about how innovation and creativity in the learning they experience will cause students to complain or provide poor evaluations or detract from the real business of research has resulted in a continued dependence on traditional models of teaching and learning and notions of success. Such models may lack authenticity, be at odds with the individual values associated with teaching and learning espoused by an individual and not provide learners with the necessary graduate attributes for the future and yet they continue. I have argued that whilst pedagogic frailty may be an understandable or inevitable consequence of a Higher Education that is dominated by neoliberal ideas of consumerism, managerialism and accountability it may contribute to a generation of graduates who are not prepared for an uncertain and unpredictable future. Now is the time for academics to rethink traditional notions of teaching and learning and how success is conceptualised so they can reverse this trend and facilitate the development of academically competent and affectively prepared graduates who can face this future with some degree of confidence. Drawing on the principles of ipsative assessment as a means to encourage In this paper I have argued current climates in Higher Education have resulted in what Kinchin (2015) calls pedagogic frailty. Academics are becoming increasingly risk- averse against a backdrop of increased marketisation and accountability. Fears about how innovation and creativity in the learning they experience will cause students to complain or provide poor evaluations or detract from the real business of research has resulted in a continued dependence on traditional models of teaching and learning and notions of success. Such models may lack authenticity, be at odds with the individual values associated with teaching and learning espoused by an individual and not provide learners with the necessary graduate attributes for the future and yet they continue. I have argued that whilst pedagogic frailty may be an understandable or inevitable consequence of a Higher Education that is dominated by neoliberal ideas of consumerism, managerialism and accountability it may contribute to a generation of graduates who are not prepared for an uncertain and unpredictable future. Conclusion Now is the time for academics to rethink traditional notions of teaching and learning and how success is conceptualised so they can reverse this trend and facilitate the development of academically competent and affectively prepared graduates who can face this future with some degree of confidence. Drawing on the principles of ipsative assessment as a means to encourage learners not simply to think about the products of learning but about the process of learning and how they experience it offers an alternative way to think about success. Success becomes an amalgam of academic and affective attributes and as a consequence learners are less intimidated by encounters with troublesome knowledge, they have learned to accept that such encounters offer challenges yes, but that they can be mastered with effort and resilience. 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Winstone (Eds.). Pedagogic Frailty and Resilience in the University . Rotterdam: Sense Publishers. Woodall, T., Hiller, A., & Resnick, S. (2014). Making sense of higher education: Students as consumers and the value of the university experience. Studies in Higher Education, 39(1), 48-67.	48,538
https://openalex.org/W4242621439	OpenAlex	Open Science	CC-By	2,021	River on Fire: The mache parapotamios and Ecological Crises in Greco-Roman Epic	Alice Fairley	English	Spoken	37,058	64,029	RIVER ON FIRE The mache parapotamios and Ecological Crises in Greco-Roman Epic By Alice Fairley Cover image: Johann Balthasar Probst, Achilles Fights by the River ii Abstract The mache parapotamios, or river battle, briefly brings nature to the forefront of epic narrative and provides an insight into perceptions of the environment. This type scene, appearing in a number of extant epics including Homer’s Iliad, Silius Italicus’ Punica, and Statius’ Thebaid, demonstrates that these poems are aware of the importance of nature as divine and cosmological, and are concerned with its relationship to humankind. The mache parapotamios does not, however, communicate a ‘green’ message in which nature is considered, or cared for, as an entity unto itself. Destruction of the environment is frequently slated as sacrilegious, and it is often equated with cosmological disorder. What is more, the narrative promotes the consensual domestication of nature so that it will benefit humankind, recognising that undamaged and benevolent nature is better than its wild and aggressive counterpart. In this way, concern for the preservation of nature in epic is anthropocentric. Using a schema with which to analyse nature in epic, we can categorise aspects of nature into domesticated or undomesticated, and natural or non-natural. In the case of the mache parapotamios, this schema takes into account the personification of the river, as well as its relationship with humans. Alliances with humans demonstrate domestication, such as the Scamander’s cooperation with the Trojans, while enmity towards humans demonstrates a lack of domestication, as when the Trebia of the Punica assaults Scipio. Furthermore, a river is natural when it is in control of itself and acting according to its phusis, but an external force, such as pollution or obstruction, can cause a river to become non-natural. This frequently reflects negatively on the human perpetrators of the non-natural phenomena. Each of the three chapters discusses one of the texts above and analyses the mache parapotamios, as well as other scenes involving nature, using the proposed schema. While each text presents an altered version of the river battle in order to best suit the needs of the epic, the significance of the relationships between the gods, humans, and nature remains a constant across all three. iii To the Water Protectors Mni Wiconi. Water is Life. E rere kau mai te Awa nui Mai i te Kāhui Maunga ki Tangaroa Ko au te Awa Ko te Awa ko au. The Great River flows From the Mountains to the Sea I am the River The River is me. iv iv Acknowledgements I am deeply grateful to my supervisor, Dr Simon Perris, for his expertise, support, and constructive feedback over the past 12 months. My sincere thanks also to my second supervisor, Professor Arthur Pomeroy, whose knowledge of Flavian epic was indispensible to this thesis. I am thankful for the generous scholarship provided by Victoria University of Wellington, which allowed me to complete this scholarship without any financial hardship. Thanks also to the Victoria University Classics Department for fostering such a supportive, friendly environment, and to my fellow postgraduates, whose antics never fail to amuse me. Finally, and most importantly, I am grateful to my family – my parents, my grandparents, and my brother – for being my role models, and for encouraging me to work harder without losing sight of what is important to me. Not to mention for all their hours spent proofreading! And to Nellie Lyon, for her love, her kindness, and for always inspiring me to be a better person. v v vi vi 1 An episode in Tacitus’ Annales (1.79) illustrates this well: Roman senators discussed the possibility that a number of the Tiber’s tributaries be diverted in order to prevent future flooding; those whose rivers would be affected argued strongly against the plan, citing dangers to their own communities and the disrespect it would show to rivers whom they had provided with rites, altars and groves. See Campbell (2012) 118-9. 6 E.g. Epictetus, Gnomologium 3.1. 7 Table of Contents Abstract……………………………………………………………………………….iii Acknowledgements……………………………………………………………………v Introduction……………………………………………………………………………1 1. Iliad 21: Achilles and the Scamander………………………………………………8 2. Punica 4: Scipio and the Trebia…………………………………………………...28 3. Thebaid 9: Hippomedon and the Ismenus…………………………………………50 Conclusion……………………………………………………………………………71 Works Cited…………………………………………………………………………..77 vii viii 4 E.g. Marcus Aurelius 4.43. 5 p , g p ( ) 2 A comical example of this is Ovid’s Amores 3.6 in which the narrator comes across a river in flood and tries to mollify it with tales of amorous rivers, only to have it swell even further. 3 E.g. Herodotus on the flooding and source of the Nile (2.19-31); Pliny on the wonders of various rivers (Natural History 2.106); and Lucretius on the replenishment of river and seawater (5.261-72). 7 Campbell (2012) 64-70, especially 65. g 5 E.g. Heraclitus, who said that we cannot step into the same river twice (DK 22A6). 6 E E i t t G l i 3 1 g 5 E.g. Heraclitus, who said that we cannot step into the same river twice (DK 22A6). 6 E.g. Epictetus, Gnomologium 3.1. Introduction It is no surprise that rivers captured the imaginations of Greek and Roman writers. River water brings beauty to the landscape, the promise of life, and prosperity through fishing, trade, and irrigation. Rivers were incorporated into domestic life as places for bathing and washing, and the deities that supposedly dwelled beneath the waves were compensated with offerings.1 Undoubtedly, it was not only the tranquil image of the river that intrigued writers, but also their ability to turn wild and lethal, able to flood and churn with the same waters that had once been slow and temperate.2 Moreover, for those with scientific interests, rivers provided a fascinating and diverse subject of enquiry.3 For philosophers, rivers were symbolic; their ever-changing state resembled the unstoppable flow of time,4 the ceaseless movement of the cosmos,5 or human life itself.6 Callimachus uses the image of two rivers to distinguish between poetic genres: the grand Assyrian River represents epic poetry, while Callimachus’ own poetry is like a small, pure stream (Hymn to Apollo 108-12). Rivers were also significant markers of identity for those who lived in the vicinity of one. Some groups were named for specific rivers, such as the Cappadocians after the Cappadox and the Arevaci after the Areva.7 Individuals could also be named for rivers, and so we have Simoisius, Scamandrius, Ismene, Almo, and Galaesus, to name a few. In the Roman Empire, rivers were valued as symbols of conquest and also for their practical use; emperors portrayed themselves as masters of 1 1 rivers, able to control and work in alliance with them for the benefit of Roman citizens.8 Unlike other features of nature, such as mountains, forests, and lakes, rivers visibly move and can even change course. Such liveliness makes them an easy target for anthropomorphism. Indeed, rivers are frequently personified in literature as minor male deities. More renowned rivers are generally personified more often and are assigned certain characteristics: the Acheloüs, for example, is well known for having only one horn, the other having been broken off by Heracles;9 the Asopus is famous for challenging Zeus after he abducted the river’s daughter, Aegina;10 and the most venerated Italian river, the Tiber, is involved in a number of Roman myths. In particular, rivers sometimes play major roles in literary narratives, especially in epic. One such role is the river battle, or mache parapotamios, in which a personified river battles a powerful warrior. p 11 Quotations in Greek and Latin are taken from the Teubner text of West (2000) for the Iliad, from the Teubner text of Delz (1987) for the Punica, and from the Loeb text of Shackleton Bailey (2003) for the Thebaid. All translations are my own. p p g y Acheloüs took the form of a horned bull. The myth is described by Apollodorus (2.7.5) and by Sophocles in the Trachiniae (509-526), and later by Ovid in the Metamorphoses (9.1-97). 0 Apollodorus 3.12.6. 1 12 This thesis is primarily concerned with the Iliad, the Punica, and the Thebaid. The Dionysiaca does not receive its own chapter owing to both space constraints and to the fact that it is a late antique epic which differs significantly from the previous three in its outlook, conception, and theology. On the mache parapotamios of the Dionysiaca, see Schmiel (2003). On the Dionysiaca in general, see Shorrock (2011) and Domenico, ed. (2016). p p g y 9 Acheloüs took the form of a horned bull. The myth is described by Apollodorus (2.7.5) and 8 Ibid. 369-88. Consider also the passage in Statius’ Silvae 4.367-94 in which the Volturnus praises the emperor Domitian for taming his unruly waters. y p 10 Apollodorus 3.12.6. 11 by Sophocles in the Trachiniae (509-526), and later by Ovid in the Metamorphoses (9.1-97). 10 Apollodorus 3.12.6. y y p by Sophocles in the Trachiniae (509-526), and later by Ovid in the Metamorphos 8 Ibid. 369-88. Consider also the passage in Statius’ Silvae 4.367-94 in which the Volturnus praises the emperor Domitian for taming his unruly waters. 9 Acheloüs took the form of a horned bull. The myth is described by Apollodorus (2.7.5) and by Sophocles in the Trachiniae (509-526), and later by Ovid in the Metamorphoses (9.1-97). 10 Apollodorus 3.12.6. 11 Quotations in Greek and Latin are taken from the Teubner text of West (2000) for the Iliad, from the Teubner text of Delz (1987) for the Punica, and from the Loeb text of Shackleton Bailey (2003) for the Thebaid. All translations are my own. 12 This thesis is primarily concerned with the Iliad, the Punica, and the Thebaid. The Dionysiaca does not receive its own chapter owing to both space constraints and to the fact that it is a late antique epic which differs significantly from the previous three in its outlook, conception, and theology. On the mache parapotamios of the Dionysiaca, see Schmiel (2003). On the Dionysiaca in general, see Shorrock (2011) and Domenico, ed. (2016). p 11 Quotations in Greek and Latin are taken from the Teubner text of West (2000) for the Iliad, from the Teubner text of Delz (1987) for the Punica, and from the Loeb text of Shackleton Bailey (2003) for the Thebaid. All translations are my own. 12 This thesis is primarily concerned with the Iliad, the Punica, and the Thebaid. The Dionysiaca does not receive its own chapter owing to both space constraints and to the fact that it is a late antique epic which differs significantly from the previous three in its outlook, conception, and theology. On the mache parapotamios of the Dionysiaca, see Schmiel (2003). On the Dionysiaca in general, see Shorrock (2011) and Domenico, ed. (2016). y y p ( by Sophocles in the Trachiniae (509-526), and later by Ovid in the Metamorphoses (9. 10 A ll d 3 12 6 Introduction This type scene occurs in four extant epics: Homer’s Iliad, Silius Italicus’ Punica, Statius’ Thebaid 11 and Nonnus’ Dionysiaca.12 The Homeric instance is our earliest surviving example of the river battle, and after the Iliad, roughly eight centuries pass before the mache parapotamios re-emerges in any substantial form in our extant literature. By the end of first century CE, both Silius and Statius had published an epic containing a mache parapotamios inspired by the Homeric source model. While it is likely that poets between Homer and the Flavians made use of the type scene in works that have now been lost to us, we cannot say for certain how they may have presented it. In any case, comparative studies by scholars such as Juhnke (1972) and Chaudhuri (2014) demonstrate that the Flavian poets certainly took their inspiration for the mache parapotamios from the Iliad. 2 As a consequence of the Homeric source model, the mache parapotamios has a common structure. This is enumerated below: 1. The river is filled with corpses, blood, and equipment. 2. The river, personified, berates the warrior for polluting the water. 3. The river fights the warrior and comes close to overwhelming him. 4. At the last second, the gods intervene, rescuing the warrior and calming the river. Each epic has its own particular take on the type scene, and the details differ depending on the author, but the basic structure remains the same throughout Homer, Silius, and Statius. For this reason the mache parapotamios makes for a useful platform on which to study rivers in epic; the core arrangement remains constant, but the changes in detail reveal the purposes for which rivers are used. Moreover, rivers are representative of nature more generally; the voices of rivers in these epics speak for other features of nature, whose lack of liveliness makes them less likely to be anthropomorphised. To help us interpret the treatment of rivers in epic, I propose a theoretical schema by which we can classify nature in ancient epic. This is expressed in the diagram below: Nature (domesticated/undomesticated) Natural Non-natural Driven by phusis. Imposed upon by external force contrary to phusis. Driven by phusis. Nature, as I refer to it, is used to describe the aggregate of all aspects of the physical world not created by humankind. This includes trees, mountains, lakes, rivers, oceans, rocks, non-human animals, and so on. Introduction A river is a part of nature, and ‘nature’ is the collective term for all these physical aspects combined. In this way I use ‘nature’ in much the same way as we use it today. Throughout this thesis I will also use ‘environment’ and ‘landscape’ as synonyms for nature. Although nature is itself not a product of human artifice, my model must also account for instances in which nature and humans interact in co-beneficial or exploitative ways. I refer to this in terms of domestication: just as there are wild 3 animals and domesticated animals, nature can likewise be divided into domesticated and undomesticated forms. We may say that domesticated nature is synonymous with activities such as agriculture, pastoralism, and hydroengineering; undomesticated nature is nature in its wild, at times uncooperative, form. As a result, domesticated nature is beneficial to humankind, while undomesticated nature poses a threat. This idea is present in our three epics, where a personified aspect of domesticated nature is loyal to its domesticators and desires to help them, while simultaneously wishing to harm their enemies, by whom it is not domesticated. The Scamander, for example, rescues dying Trojans while attacking Achilles. In epic, therefore, the status of nature as either domesticated or undomesticated depends both on an anthropocentric perspective on its relationship to humankind, and on the specific details of its role in the narrative. An aspect of nature may be domesticated by a specific group, but to other groups that same aspect may appear undomesticated. According to my schema, nature can be divided into two subsets: ‘natural’ and ‘non-natural’ nature. The two subsets of nature refer not to an object’s materiality, but rather to its innate motivating force, which I call phusis. I borrow this term from pre- Socratic philosophy, in which phusis is the source of an entity’s behaviour, the reason for why it acts in the way it does, without pressure or restraint.13 For the pre- Socratics, phusis was an identifiable element, and each philosopher had a different idea as to what that element was. Thales believed that it was water (DK 11A12), while Heraclitus thought it more likely to be fire (DK 22B30). Anaximander, on the other hand, took a more abstract approach, believing the principle to be τὸ ἄπειρον, or the ‘Boundless’, which is infinite both in space and time (DK 13A5, B1, A11, A17). 13 Collingwood (1945) 44. This was, at least, one interpretation of phusis, though Naddaf (2005) 11-35 explains that the pre-Socratic understanding of phusis was much more nuanced than one definition would allow. Rather, it could refer to primordial matter (such as fire, water, etc.), intrinsic force that he calls ‘process’, and/or the result of a combination of both. I have chosen to use the term phusis in terms of an intrinsic force, as per Collingwood’s interpretation. p 14 E.g. Anaximander (DK 13A5): οὗτος ἀρχὴν ἔφη τῶν ὄντων φύσιν τινὰ τοῦ ἀπείρου, ἐξ ἧ γίνεσθαι τοὺς οὐρανοὺς καὶ τὸν ἐν αὐτοῖς κόσµον. 15 14 E.g. Anaximander (DK 13A5): οὗτος ἀρχὴν ἔφη τῶν ὄντων φύσιν τινὰ τοῦ ἀπείρου, ἐξ ἧς γίνεσθαι τοὺς οὐρανοὺς καὶ τὸν ἐν αὐτοῖς κόσµον. 15 Naddaf (2005) 11-35. 16 There exists an adjective phusikos which may be translated as ‘natural’, or ‘inborn’. Aristotle employs this term (e.g. in Physica 191a3), but it is not found in the pre-Socratic fragments. Introduction Moreover, the pre-Socratics thought that the primary element was from which everything else was created,14 and that it therefore dictated the movements of the cosmos, as well as everything within the cosmos. Accordingly, phusis was commonly associated with growth and creation.15 4 Returning to our key terms, ‘natural’ and ‘non-natural’, we can now say that that which acts according to its phusis is natural.16 Conversely, that which is forced to act or exist against its phusis is non-natural.17 For example, a river that flows from spring to sea is acting according to its phusis in a natural manner. But a river that is forced to flow backwards, from sea to spring, is acting in a way that is contrary to its phusis and therefore non-natural.18 In this way, natural and non-natural states are not exclusive to aspects of nature. That which is ‘natural’ is not always a part of ‘nature’; man-made objects have phuseis of their own, and can therefore be natural or non- natural too. Such a framework is all well and good, but without a concrete means of identifying which subset is which, it is not all that useful. With the addition of personification, this becomes possible. In the three epics to be discussed, the rivers involved in the mache parapotamios all experience non-natural phenomena; they are also all personified. As anthropomorphised entities, the rivers are able to express distaste for certain experiences; for example, all three lament the inundation of corpses that pollutes their streams. Because an external force causes the inundation, and because the river makes clear that it is not something that it desired or chose, we can conclude that the corpse-filled river is a non-natural phenomenon. Volition, and therefore personification, is key in deciding whether an event is natural (desired) or non-natural (undesired). However, to make matters easier, there are three phenomena that are almost always non-natural in the mache parapotamios. These are: the corpse- filled river, the obstructed river, and the backwards-flowing river. Using this schema, I analyse the treatment of the riverine environment and other relevant aspects of nature in each of the three texts. By classifying domesticated and undomesticated nature, and interpreting the effects of non-natural phenomena on nature, I identify a number of implicit attitudes towards the landscape. g 17 Note that I have chosen to use the term ‘non-natural’ rather than ‘unnatural’. This was a conscious choice intended to prevent normative associations. Nevertheless, as we proceed we will find that non-natural phenomena are indeed often portrayed as morally repugnant. p p y y p g 18 Note that a non-natural action or state is always provoked by an external force. 23 E.g. the Five Ages of Man myth as described in Hesiod’s Works and Days: in the Golden Age humankind is uncivilised, but nature provides all that is needed for survival of its own accord. By the Iron Age, humankind has gained civilisation, but nature has grown wild and does not reproduce unless humans constantly interfere, a wearisome burden that marks the Iron Age as the worst out of the five. See also the Myth of the Ages in Virgil, Georgics 118- 59, in which Jupiter ends the Golden Age and introduces labor improbus (145-6). p g p ( ) 24 That is, concerned with preserving nature for the sake of nature, not for the benefit of the gods or humans. g ( ) 21 E.g. the Enuma Elish, the Epic of Gilgamesh, the Labours of Heracles, Cadmus and the Serpent of Ares, Theseus and the Minotaur, etc. 22 p 22 Hughes (1994) 32-7. Without agriculture and other practices, cities could never have developed. 20 E.g. Oenotria Tellus of the Punica (15.522). 21 g ( ) 20 E.g. Oenotria Tellus of the Punica (15.522). 21 g ( ) 21 E.g. the Enuma Elish, the Epic of Gilgamesh, the Labours of Heracles, Cadmus and the Serpent of Ares, Theseus and the Minotaur, etc. 22 Hughes (1994) 32-7. Without agriculture and other practices, cities could never have developed. 23 19 Hughes (1994) 46. 20 19 Hughes (1994) 46. 20 E.g. Oenotria Tellus of the Punica (15.522). 21 E.g. the Enuma Elish, the Epic of Gilgamesh, the Labours of Heracles, Cadmus and the Serpent of Ares, Theseus and the Minotaur, etc. 22 Hughes (1994) 32-7. Without agriculture and other practices, cities could never have developed. 23 E.g. the Five Ages of Man myth as described in Hesiod’s Works and Days: in the Golden Age humankind is uncivilised, but nature provides all that is needed for survival of its own accord. By the Iron Age, humankind has gained civilisation, but nature has grown wild and does not reproduce unless humans constantly interfere, a wearisome burden that marks the Iron Age as the worst out of the five. See also the Myth of the Ages in Virgil, Georgics 118- 59, in which Jupiter ends the Golden Age and introduces labor improbus (145-6). 24 That is, concerned with preserving nature for the sake of nature, not for the benefit of the gods or humans. 19 Hughes (1994) 46. 20 1 Various explanations have been proposed for this disjunction: Leaf (1902) 401 considers the possibility of a ruse on the river’s part, or that we should read Scamander’s plea and Achilles’ response as irony; Willcock (1984) 287 suggests that Achilles’ assent may be much less clear than it appears. Less psychologizing interpretations are proposed by Richardson (1993) 70, who tentatively suggests there may have been another version that did not include the river battle, and West (2011) 378, who describes the transition as “bafflingly illogical,” suggests that line 233 was added later and integrated unsuccessfully. Introduction These are primarily religious and cosmological; both the Greeks and Romans thought nature to be closely connected to the presence and actions of the gods, and occurrences in the 5 natural world were considered realisations of divine acts.19 Furthermore, features of nature could have divine lineage and be divine themselves; Hesiod, for example, claims that rivers are children of Tethys and Oceanus (Theogony 337-45). Aspects of nature were often personified as gods or were closely connected to specific divinities: for example, Gaea the earth goddess, and Oceanus the ocean boundary and father of rivers. Rivers had their own personifications, as did the four winds, and occasionally entire lands could be a personified as a single deity.20 Also personified were some planetary bodies and phenomena: Helios (sun) and Selene (moon), as well as the dawn goddess, Eos. Meteorological phenomena were also thought to be of divine origin, usually of Zeus/Jupiter’s doing. However, of equal concern is the interaction between humans and nature. While many myths describe the tension between nature and civilisation,21 with civilisation the unequivocal victor, there remains the inescapable truth that civilisation cannot develop without nature. 22 The need for domesticated nature is often acknowledged, although interaction with nature is frequently seen as undesirable.23 Nevertheless, implicit in the three epics is a preference for domesticated nature over undomesticated nature, as the former may be beneficial to humans, while the latter is a potential threat. However, owing to the potential for any aspect of nature to be numinous, the morality of domestication is often ambiguous; this issue is explored in several of the epics discussed in the chapters to follow. There is no evidence that any of the texts ascribe value to nature as a thing unto itself. Nature derives its value from its divinity and its relationship with humankind. Therefore any concerns regarding pollution and obstructions are not what we might call environmental.24 Rather, they are, as outlined above, overtly religious 6 and cosmological, and implicitly anthropocentric concerns. Nevertheless, as this thesis will show, these texts demonstrate a general appreciation for nature as a significant physical presence in the world, and an acknowledgement of the fact that the condition of the landscape directly influences human activities, sometimes in ways that cannot be controlled. 7 1. Iliad 21: Achilles and the Scamander The Iliad describes a brief period during the final year of the Trojan War, and focuses primarily on Achilles, as well as other Greek and Trojan warriors. Within that timeframe Achilles retires from the fighting, only to return to battle after the death of his friend, Patroclus. He kills the Trojan hero, Hector, whose burial takes up the final episode of the epic. This chapter is concerned with the first half of book 21, in which Achilles battles the River Scamander. An outline of this passage is given below. Achilles, roused to a frenzy following the death of Patroclus, drives a great many Trojan soldiers into the Scamander River where he slaughters them indiscriminately. As a result of his rampage, the river becomes clogged and bloody with corpses: πλήθει γὰρ δή µοι νεκύων ἐρατεινὰ ῥέεθρα, \| οὐδέ τί πῃ δύναµαι προχέειν ῥόον εἰς ἅλα δῖαν (“my lovely streams are full of corpses, and I cannot pour forth my water into the holy sea,” 21.218-9). In response the Scamander appears in a man’s form (ἀνέρι εἰσάµενος, 213) and rebukes Achilles for his vicious treatment of the Trojans and for disrespecting the river. The Scamander demands that Achilles take the fighting elsewhere and leave its stream alone: ἐξ ἐµέθεν γ᾽ ἐλάσας πεδίον κάτα µέρµερα ῥέζε (“drive [the Trojans] away from me and onto the plain, and there perform your baneful deeds,” 217). Achilles says he will relent, but soon returns to battle the river directly.1 Their fight continues for 94 lines (233-327), in which the Scamander comes very close to overwhelming Achilles with huge waves and strong currents. When the situation appears most dire, Hera steps in and orders Hephaestus to send his fires against the Scamander (328-41); these successfully quell the river’s fury and it agrees to retire from the fighting (372-6). The description of the fight between Achilles and the Scamander is one of the few scenes in the Iliad that sets nature directly into the narrative. Elsewhere nature is almost exclusively reserved for similes, epithets, or passing mentions of distant 8 homelands.2 Moreover, the mache parapotamios does not present nature as a benign object of contemplation, but a wilful participant in the action; the personification of the Scamander allows it to enter the narrative as a physical character whose presence and movements are deliberate and consequential. Cyclopes (9.116 41); Scylla s cave (12.73 85); Ithaca (13.236 49), etc. 3 E.g. anger: χωσάµενος (21.212), κεχόλωτο (146), χολώσατο (136); pity: (145-7). 4 2 Compare the many lengthy descriptions of nature in the Odyssey: Calypso’s island (5.55- 74); the Phaeacian shore where Odysseus spends a night (5.451-85); the island of the Cyclopes (9.116-41); Scylla’s cave (12.73-85); Ithaca (13.236-49), etc. g g χ µ ς ( ), χ ( ), χ ( ); p y ( ) 4 See Yamagata (1994) 180. g g χ µ ς ( 4 See Yamagata (1994) 180. 1. Iliad 21: Achilles and the Scamander It, like other men and gods, feels anger and pity and acts on these emotions.3 As outlined in the introduction, nature may be divided into two categories: domesticated and undomesticated. When the river actively aids the Trojans and does so deliberately, it shows itself to be domesticated. Thus, when the river revives Hector at 14.433-9, allows women to wash their clothes in its stream (22.153-6), and hides Trojan soldiers beneath its waves (21.238-9), it does so because it is loyal to the community that has domesticated it, pities their suffering, and wishes to help and protect them. Nevertheless, the poet reconciles this mild and pleasant river with one of the realities of nature: that it is not always so temperate, and can be as contrary as it is cooperative. And uncooperative nature is, at least within the realms of epic, undomesticated. As the river’s anger rises, it begins to oppose the human force that it views as a threat, by appearing as a man to address and rebuke Achilles directly, and by attacking the Greek warrior with massive waves and flooding (21.234-71). These actions, though irregular and excessive, remain natural, as they are expressions of the river’s volition. The anger and violence of the river directed at Achilles is both a reflection of nature when it is undomesticated, and simultaneously a continuation of Trojan domestication over it, since a source of the river’s anger is that Achilles does not pity the suffering Trojan warriors: µένος δέ οἱ ἐν φρεσὶ θῆκεν \| Ξάνθος, ἐπεὶ κεχόλωτο δαῒ κταµένων αἰζηῶν, \| τοὺς Ἀχιλεὺς ἐδάϊζε κατὰ ῥόον οὐδ᾽ ἐλέαιρεν (“Xanthus put strength in his [Asteropaeus’] heart, enraged because of the youthful men killed in battle, whom Achilles had slain in his stream without pity,” 145-7).4 It is essential to consider the river’s actions in terms of phusis, which propels it to act in certain ways. Whatever the river does willingly is done according to the river’s phusis. If the river is domesticated, then it is the phusis of the river to be protective and sympathetic to its domesticators; if undomesticated, it is its phusis to be threatening and defensive. Homer demonstrates that the phusis of the river always 9 9 remains potentially variable, that is, it always holds within it the possibility of being either domesticated or undomesticated. g ( ) 8 Indeed Achilles and his companions appear to experience no lack of food or wine (e.g. 9.205-28; 24.621-30). Note a few interesting mentions of consumption: the wine sale at the end of book 7 (467-82) in which the Achaean soldiers purchase wine for a great feast. Notably, some soldiers pay for their wine with cattle. Also Odysseus’ words to Achilles at 19.160-70 acknowledge that soldiers need food and water. Then at 24.778-84 Priam orders the Trojans to collect firewood for Hector’s pyre, and they carry back an unspeakably great (ἄσπετον) amount (784). For more on this, see Bakker (2010) 48-50. 5 In fact, two agrarian similes feature in the river battle episode: Achilles is compared to a farmer whose ditch is overtaken by running water (21.257-64) and Hephaestus’ fires are compared to the North Wind, which dries a field so it can be tilled by the farmer (346-7). Notable too are the descriptions of nature found on the shield of Achilles (18.483-608). p ( ) 6 Taplin (1980) 15, in his discussion of the shield of Achilles, asks us to consider the Iliadic similes and the images on the shield as windows into the world behind the war, one of peace, agriculture and craftsmanship, to where the soldiers will likely never return: “the similes make us see war as wasteful and destructive.” See also Redfield (1975) 186-192. 1. Iliad 21: Achilles and the Scamander Thus the Scamander can protect the Trojans beneath his waves and use those same waves to attack Achilles, all in a single motion (238-42). The presentation of domesticated nature in the Iliad is a relatively unambiguous one. The war setting entails that human interactions with nature are few, and usually only represented in similes or ekphrasis.5 As a result, agricultural images are generally depicted as positive and productive; they represent the antithesis of war.6 Thus the domestication of nature is viewed as something desirable, as Jonathan Burgess notes: “The Iliad and the Odyssey support anthropocentric exploitation of natural resources.”7 Nevertheless, it is clear that there is little place for domesticated nature on the battlefield: the need for renewable resources is not acknowledged as an issue, despite ten years of incessant warfare,8 and the mechanisms of war are much more frequently compared to the movement of undomesticated nature.9 Thus soldiers are often compared to rushing torrents of water or flash floods (5.85-92; 16.384-93; 17.263-6, etc.), which actively destroy the agricultural works of humans: dams, dikes, fields, fences and orchards are all flattened beneath the onslaught.10 Domesticated nature is shown to be weak in the face of its wild counterpart, and when the domesticated Scamander tries to protect those people to whom he is loyal, he fails them. This is because the war of the Iliad does not accommodate human/nature g ( ) 8 Indeed Achilles and his companions appear to experience no lack of food or wine (e.g. 9.205-28; 24.621-30). Note a few interesting mentions of consumption: the wine sale at the end of book 7 (467-82) in which the Achaean soldiers purchase wine for a great feast. Notably, some soldiers pay for their wine with cattle. Also Odysseus’ words to Achilles at 19.160-70 acknowledge that soldiers need food and water. Then at 24.778-84 Priam orders the Trojans to collect firewood for Hector’s pyre, and they carry back an unspeakably great (ἄσπετον) amount (784). For more on this, see Bakker (2010) 48-50. 9 E.g. the simile at 17.53-69, which pits a domesticated olive tree directly against the undomesticated and tempestuous winds. 10 9 E.g. the simile at 17.53-69, which pits a domesticated olive tree directly against the undomesticated and tempestuous winds. 10 10 Note that it is primarily the Trojans who are compared with rivers and other freshwater sources. 7 Burgess (2015) 113. 8 11 Yamagata (1994) 40-5; Chaudhuri (2014) 198: “Achilles’ dismissal of the Trojans’ worship of their local river deity as ineffective undermines the system of hoped-for reciprocity underlying many of the prayers within the poem.” Consider especially the failed supplication of Lycaon at 21.71-119. g ( ) 15 For more on winds in the Odyssey (and the Iliad) see Purves (2010) 323-50. y 12 Johnston (1988) 31-54 argues that by comparing soldiers to wild natural phenomena, Homer implies that warfare is itself a part of nature: “warfare is thus not a human aberration but an integral part of the irresistible, eternal, and mysterious natural order of things” (33). 13 Although it is not clear that the authors of the Iliad and the Odyssey are a single unit, they h l f i di i d h f h i g p y g ( ) 13 Although it is not clear that the authors of the Iliad and the Odyssey are a single unit, they are nevertheless part of a contemporary epic tradition and therefore worth comparing. 14 p p y p 14 Burgess (2015) 113. p 14 Burgess (2015) 113. 15 y 2 Johnston (1988) 31-54 argues that by comparing soldiers to wild natural phenomena, Homer implies that warfare is itself a part of nature: “warfare is thus not a human aberration but an integral part of the irresistible, eternal, and mysterious natural order of things” (33). Homer implies that warfare is itself a part of nature: warfare is thus not a human aberration but an integral part of the irresistible, eternal, and mysterious natural order of things” (33). 13 Although it is not clear that the authors of the Iliad and the Odyssey are a single unit, they are nevertheless part of a contemporary epic tradition and therefore worth comparing. 14 Burgess (2015) 113 1. Iliad 21: Achilles and the Scamander The Greeks are far more often compared with the sea: see Fenno (2005). Animal and meteorological similes are also common. 10 Note that it is primarily the Trojans who are compared with rivers and other freshwater sources. The Greeks are far more often compared with the sea: see Fenno (2005). Animal and meteorological similes are also common. 10 alliances, much as it does not accommodate the fulfilment of other peacetime customs such as supplication, a point I will return to later.11 Although pastoral and agricultural interaction with nature is presented as something positive, it is nevertheless part of an entirely separate world. For this reason, the moral ambiguities surrounding environmental exploitation are not at issue, and the text presents a relatively black- and-white view of human involvement in nature: none of the similes or descriptions shows the destructive potential of domesticating nature. Instead the dangerous aspects of nature are emphasised, suggesting that undomesticated nature is much more at home in the world of war; nothing equals the violence of the battlefield like nature in full, unrestrained force.12 In the Odyssey there is more room for tension with regards to the environment.13 Where the Iliad places nature outside the physical setting at Troy, the Odyssey brings it back into the central narrative, using descriptions of nature to elicit a sense of space and geography. However, unlike the Iliad, where the threat of undomesticated nature is suitably related to the soldiers clashing on the battlefield, the Odyssey depicts a world at peace, one that is at odds with undomesticated nature. Here, undomesticated nature is a threat to the new, peaceful order of things; it manifests physically as monstrous creatures and dangerous characters, such as Scylla, Charybdis, and the Cyclopes.14 Conversely, the domestication of nature is venerated: when Odysseus takes from Aeolus the domesticated winds, he and his men sail home with ease, but upon releasing the winds, i.e. ‘undomesticating’ them, they are blown off course and must begin all over again (10.14-55).15 Also note the interaction between Odysseus and the Phaeacian river, in which Odysseus appeals to the river’s sympathies by supplicating it, successfully winning over the river’s loyalties and convincing it to carry him to safety (5.441-53). 16 Hughes (1994) 53. 1 g ( ) 17 Yamagata (1994) 28; Boyd (2009) 306-7 Hughes (1994) 53. 17 Yamagata (1994) 28; Boyd (2009) 306-7 6 Hughes (1994) 53. 7 (1994) 28 d (2009) 306 1. Iliad 21: Achilles and the Scamander Other passages hint at the moral duty 11 towards pastoralism, agriculture and fishing, such as Odysseus’ response to Penelope’s questioning in book 19: τὴν δ᾽ ἀπαµειβόµενος προσέφη πολύµητις Ὀδυσσεύς: ‘ὦ γύναι, οὐκ ἄν τίς σε βροτῶν ἐπ᾽ ἀπείρονα γαῖαν νεικέοι: ἦ γάρ σευ κλέος οὐρανὸν εὐρὺν ἱκάνει, ὥς τέ τευ ἢ βασιλῆος ἀµύµονος, ὅς τε θεουδὴς ἀνδράσιν ἐν πολλοῖσι καὶ ἰφθίµοισιν ἀνάσσων εὐδικίας ἀνέχῃσι, φέρῃσι δὲ γαῖα µέλαινα πυροὺς καὶ κριθάς, βρίθῃσι δὲ δένδρεα καρπῷ, τίκτῃ δ᾽ ἔµπεδα µῆλα, θάλασσα δὲ παρέχῃ ἰχθῦς ἐξ εὐηγεσίης, ἀρετῶσι δὲ λαοὶ ὑπ᾽ αὐτοῦ (19.106-14). Then the wily Odysseus said in reply: “woman, no mortal on this boundless earth could quarrel with you. For your fame reaches the wide heaven, just as the fame of a noble king does, who, god-fearing, lords over many strong men and maintains justice. The black earth produces wheat and barley, the trees are laden with fruit, the sheep reproduce unceasingly, and the sea provides fish from his good leadership, as his men thrive under him.” In this passage, fertility of the earth and sea is directly connected to good leadership.16 On the other hand, excessive exploitation of the land and home is a sign of moral failure, as when the suitors devour the products of Odysseus’ oikos: ἀλλὰ ἕκηλοι \| κτήµατα δαρδάπτουσιν ὑπέρβιον, οὐδ᾽ ἔπι φειδώ. \| ὅσσαι γὰρ νύκτες τε καὶ ἡµέραι ἐκ Διός εἰσιν, \| οὔ ποθ᾽ ἓν ἱρεύουσ᾽ ἱερήϊον, οὐδὲ δύ᾽ οἴω: \| οἶνον δὲ φθινύθουσιν ὑπέρβιον ἐξαφύοντες (“But at ease they greedily consume our provisions, with no thought for sparing it. For as many days and nights that come from Zeus, they sacrifice not just one or two animals. And they waste the wine, drawing it forth greedily,” 14.91-5).17 However, the domestication of nature is at times problematized, as the episodes involving the Lotus-Eaters and Circe demonstrate. In the land of the Lotus- 12 Eaters, men consume the lotus plant and afterwards forget all knowledge of their previous lives (9.82-104).18 Here the relationship between humans and nature does not result in a positive outcome, rather it serves to disrupt the normal functioning of civilisation. Similarly, when Odysseus’ men first encounter Circe she transforms them into pigs (10.237-43). 18 The exact identification of the lotus plant is unknown. Herodotus (4.177) describes it as a similar in size to the mastic-berry with a sweet taste like a date. Polybius (12.2) describes it further, and claims it was harvested and ground up or pitted and preserved for food; it could also be used to make wine and vinegar. 19 g ( ) 20 Austin (1975) 144-5. 19 Burgess (2015) 113-4. g 19 Burgess (2015) 113-4. 20 Burgess (2015) 113 4. Austin (1975) 144-5. 1. Iliad 21: Achilles and the Scamander Circe ‘domesticates’ other wild creatures using potions (212-9) and her very servants are the children of nature: γίγνονται δ᾽ ἄρα ταί γ᾽ ἔκ τε κρηνέων ἀπό τ᾽ ἀλσέων \| ἔκ θ᾽ ἱερῶν ποταµῶν, οἵ τ᾽ εἰς ἅλαδε προρέουσι (“They were born from springs and groves, and from the holy rivers that flow towards the sea,” 350-1). But Circe’s domestication of nature is disruptive and threatening; it does not have a positive impact on human society and verges on the non-natural. Burgess proposes that such episodes as these indicate a “negative valuation of [nature],”19 but other more positive depictions suggest that the perception of nature in the Odyssey is much more ambiguous. While the domestication of nature can be difficult and yield horrifying results, when done correctly it is a boon for civilisation. Attention should also be given to the unique position of the Cyclopes and the Phaeacians. Both of these groups are associated with the Golden Age, and this is reflected in their relationship with nature. The Cyclopes, while avid pastoralists, do not practice any kind of agriculture. Yet the earth provides all they need without any interference (9.106-11). The Cyclopes’ lack of agricultural endeavour is emphasised by the description of the neighbouring island. This place, inhabited only by goats, is not very far by ship from the island of the Cyclopes, but its soil is perfect for growing food and there is a supply of fresh water, as well as a suitable harbour (116-39). Despite the Cyclopes’ proximity to this arable land, they do not have the ships needed to get there (125-30), nor, presumably, the skills to cultivate it for civilisation.20 The Cyclopes’ relationship with nature indicates that they linger in the Golden Age, when people had no need for agriculture because the earth reproduced of its own accord (Hesiod, Works and Days 116-20). However, the barbarity of the Cyclopes puts the regular fecundity of the earth in an ominous light: their community (if it can be called 13 that) is highly uncivilised, and they lack the necessary motivation to form an appropriate society. The Phaeacians are much closer to civilisation than the Cyclopes are; yet they hover on the threshold of the Golden Age. It is said that they are dear to the gods (6.203) and they live far from where men toil for a living (8). 21 Perhaps their previous homeland was in fact Goat Island. See Clay (1980) 263-4 for such an argument. 22 g 22 Dougherty (2001) 91-2. 1. Iliad 21: Achilles and the Scamander Furthermore, they once lived near the Cyclopes, that other Golden Age race (5).21 Like the Cyclopes, the Phaeacians inhabit a land that provides bountiful crops without need for agriculture; for example, Alcinous’ palace gardens yield fruit all year round (7.114-21). Nevertheless, there is some mention of tilled fields (ἀγροὺς... ἔργ᾽ ἀνθρώπων, 6.259) so Phaeacia is a liminal setting that lingers between the Golden Age and what Dougherty dubs the “New World.”22 Indeed, Phaeacia is Odysseus’ final stop, from where he makes the leap out of the fantastic world of the Lotus-Eaters, Circe, the Cyclopes and Calypso, into the civilised world of Ithaca. Though the Phaeacians are decent, peaceful people, their perpetual prosperity must eventually come to an end when they incur the wrath of Poseidon by helping Odysseus return home (13.128-38), serving as a reminder that the people of the Golden Age are incompatible with the world that Odysseus belongs in. In both of these examples, nature is neither undomesticated nor domesticated, at least in the terms I have outlined. Yet Homer makes clear that this kind of relationship with nature is an echo of the past, reserved only for those who are part of a previous, and now waning, age. It should be clear at this point that the depiction of nature in the Odyssey is far more multifaceted than that in the Iliad. The Iliad presents nature as largely absent from the narrative, and domestication of it is desirable but unrealistic in the wartime setting; undomesticated nature is generally presented in martial similes, and though threatening, is rarely monstrous. Conversely, the Odyssey proposes limitations on the domestication of nature: it presents domestication as a reality, but one that is not always unambiguously positive. On the other hand, undomesticated nature is out of place in the peacetime setting, and is represented by threatening creatures like Scylla and Charybdis. 14 14 So far I have discussed those actions that the river performs willingly, and these I have labelled as ‘natural’ insofar as they are products of the river’s volition and therefore its phusis. However, there are at least two notable phenomena that are caused by external motivation: the first is the filling of the water with blood, corpses and equipment (21.235-8; 316-8; 325) and the second is the choking of the river with corpses and the subsequent obstruction of its flow (218-9). 23 West writes: “The choking of the stream with corpses provided a naturalistic explanation of the flood”. I acknowledge that the river’s reaction to the obstruction does follow “physical laws,” in the sense that it does what one would expect a real river to in such a situation, i.e. it floods. However, I view this specific event as non-natural for the reasons outlined above. 24 Mackie (1999) 493. Ibid. especially 497 8. 26 Redfield (1975) 251. That is, polluted in the physical sense; miasma is not necessarily present here, as Parker (1983) 66 notes. 27 Jones (2005) 23-4. ( ) 25 Ibid. especially 497-8. b d. espec a y 97 8. 26 Redfield (1975) 251. That is, polluted in the physical sense; miasma is not necessarily present here, as Parker (1983) 66 notes. 1. Iliad 21: Achilles and the Scamander These I call ‘non-natural’ as they affect the river in ways that hinder the river’s phusis and frustrate the river’s will.23 Furthermore, these events are caused by human actions; they are by-products of human/human interaction, rather than deliberate attacks by humans against nature. The Scamander, to whom is given a wide range of attractive epithets, including ‘fair flowing’ (14.433; 21.1; 130; 24.692), ‘deep-eddying’ (15), and ‘silver- eddying’ (8; 130),24 is transformed into a cesspool of blood and gore by these non- natural events. On a very basic level, nature is affected by war aesthetically: the carnage of the battle turns a beautiful stream into something ugly. Furthermore, Achilles’ goading of the river compels it to swell and flood, an image that is at odds with the mild river found elsewhere in the epic. The aesthetic transformation reaches its apex when Hephaestus sets the river alight, perhaps in reference to the Underworld’s own burning river, the Pyriphlegethon.25 Of primary concern are the non-natural events described above; this is because they have potentially widespread consequences that extend beyond the world of nature. When the river fills with blood and other contaminants it becomes polluted and loses its purifying properties.26 Where once the women of Troy washed their clothes is now a river filled with dead bodies and blood – it could hardly be considered a suitable place for washing and bathing now. Furthermore, the Scamander cleanses the dead, but in doing so it becomes polluted itself.27 In its polluted state the Scamander lacks the very force of life – pure water – that sustains the people who rely on it. However, matters are even worse than this. A polluted river, if unobstructed, 15 will eventually carry the filth out to sea where it may disperse, and the waters will once again run clear. In the case of the Scamander, the piling up of bodies has blocked its flow, forcing corpses to stagnate where they fall; the river is stuck, its only choice to heave the bodies up onto the earth beside it (21.235-8). Thus we observe that through the non-natural event of human-induced pollution, the river’s phusis, to act as a purifier, is hindered. Furthermore, the river’s phusis dictates that it must flow out to sea, yet the blockage in its stream prevents it from doing so. ( ) 29 E.g. the surging Greek assembly (2.144-6); the roaring of the Greeks like the surf (2.394-7); Greeks advancing to battle like waves upon the shore (4.422-8); the rushing (2.809-10), pouring (12.470; 15.360) and rumbling (2.810) of the Trojans. y 1 Rivers were commonly used as markers of place and identity in Greece, owing to their unction as boundaries and givers (or takers) of life: see Huskinson (2005) 248-9. 2 p g ( ; ) g ( ) j 30 For a complete list, see Fenno (2005) 483, n. 21. Most notable, perhaps, is Hector’s own son, Astyanax, also known as Scamandrius. 31 g ( 32 Mackie (2008) 183, also (1999) 493. 32 Mackie (2008) 183, also (1999) 493. 28 Fenno (2005) 475-504. 28 Fenno (2005) 475-504. 1. Iliad 21: Achilles and the Scamander Nature, here represented by the river, is transformed into something unfamiliar, but the effects of this transformation penetrate the layers of human society also. To understand why this is so, let us turn to the relationship between the Trojans and the Scamander. Fenno has noted that the Trojans are associated closely with fresh water, while the Greeks are linked with salt water.28 This is most clearly evident in Homer’s similes, which liken Greek soldiers to the sea and Trojans to rivers.29 However, the Trojans are further associated with rivers through their names and heritage; Asteropaeus is the grandson of the Axius River, while many other Trojans are named after rivers.30 It is clear that Trojan identity is closely linked with rivers, and most prominently with the Scamander.31 Their identities are so tied together, in fact, that the mache parapotamios episode directly foreshadows the future destruction of Troy. Mackie expands upon this idea, noting that although Troy is never razed within the confines of the Iliad, the city is, nevertheless, destroyed in other ways, namely through other fire-related events, such as the burning of the Scamander and the cremation of Hector.32 Indeed, the river itself assures Hera that it will no longer serve as the protector of Troy after it is set aflame: ἐγὼ δ᾽ ἐπὶ καὶ τόδ᾽ ὀµοῦµαι, \| µή ποτ᾽ ἐπὶ Τρώεσσιν ἀλεξήσειν κακὸν ἦµαρ, \| µὴδ᾽ ὁπότ᾽ ἂν Τροίη µαλερῷ πυρὶ πᾶσα δάηται \| καιοµένη, καίωσι δ᾽ ἀρήϊοι υἷες Ἀχαιῶν (“and I will even swear this: I will not defend the Trojans from that evil day, not when all Troy burns with a consuming fire that the warlike sons of Achaea ignite,” 21.373-6). Fire is a potent symbol of 16 Troy’s fate, but the destruction of the Trojan city may be assured by other images too: just as the idyllic river is transformed into a hellish stream of blood, filth, death, and suffering, so too will the once-glorious Troy be reduced to a polluted ruin. In other words, the fate of the Scamander is entwined with the fate of Troy and its people, both as a symbol that represents Trojan life, and as a physical ally that provides life through its waters. We have seen how both the environment and humankind are affected by non- natural phenomena; I will now turn my attention to the effects of these events within the divine realm. g ( g y ) 34 Indeed, some versions of Diomedes’ life actually immortalize him, e.g. Pindar, Nemean 10 Διοµήδεα δ᾽ ἄµβροτον ξανθά ποτε Γλαυκῶπις ἔθηκε θεόν. 33 This contradicts Hesiod who claims that the Scamander was born of Tethys and Oceanus; nevertheless rivers in Hesiod are still of divine origin (Theogony 337-45). 34 µή µβρ ξ ς η 35 The Scamander itself notes this in its speech to Achilles: αἰεὶ γάρ τοι ἀµύνουσιν θεοὶ αὐτοί (21.215). 1. Iliad 21: Achilles and the Scamander The Scamander fulfils a variety of roles within the narrative, as a feature of the landscape and an ally to the Trojans, but it, alongside other rivers, is also an immortal. Like other gods, the Scamander has a priest (5.76-83) and is given offerings of horses and bulls (21.131-2). Rivers in general are counted among the gods in the Iliad, and they are summoned alongside all the other divinities to Zeus’ halls (20.7). Furthermore, the Scamander is a son of Zeus (21.2).33 We must therefore consider Achilles’ actions and the related non-natural phenomena as events that have the potential to elicit the ire of the gods. Indeed, we have already seen how the Scamander protests the slaughtering of the Trojans in its stream at 21.212-21 and its violent reaction to Achilles’ attack. However, the Iliad does not actively condemn mortals who fight immortals. 5.330-42 describes the wounding of Aphrodite at the hands of Diomedes, yet he receives no punishment for attacking the goddess. Instead Dione points out to Aphrodite that she is not the first divinity to be wounded by a mortal (381-4), and although she predicts death for those who battle the gods (406-9), Diomedes escapes any such fate in the Iliad.34 If we return to Achilles and the Scamander, we might note that the river does not complain that the Greek hero is sacrilegious in his actions; rather that he is tainting the water and killing too many Trojans. Furthermore, instead of being punished, Achilles actually receives help from Poseidon, Athena, Hera and Hephaestus so that he triumphs over the river.35 Much thought has been given in the scholarship to Homeric morality. Perhaps most quoted is Dodds, who noted that the gods of the Iliad do not concern themselves 17 with justice.36 Adkins, too, saw the gods of Homer as unjust.37 Since then, critics have countered this view with more nuanced arguments. In The Justice of Zeus, Lloyd- Jones proposes that the early Greek idea of justice hinged on an understanding of order, which was maintained through reciprocity of time.38 Zeus dispensed the themistes to kings such as Agamemnon, so that they might uphold the principles of reciprocal justice.39 When Homeric heroes fail to award each other the proper time, as Agamemnon refuses to respect the time of Achilles, they violate the order of justice overseen by Zeus. ( ) 38 Lloyd-Jones (1971) 4. ( ) 37 Adkins (1960) 62. 36 Dodds (1951) 32. Ibid. 7. 40 Ibid. 27: “what is just for mortals is not necessarily what mortals want.” 41 Ibid. 9-10. 42 1. Iliad 21: Achilles and the Scamander There are certain behaviours, which, if they upset the proper allotment of time, are disagreeable to the gods. However, it is not always obvious to mortals when order has been, or might be, disrupted, and for this reason the justice of Zeus does not always appear fair or equal.40 Lloyd-Jones also stresses the concept of double motivation; human characters of the Iliad are always responsible for their actions, even if the gods directly influence those actions.41 Kullman agrees that the gods operate according to the principles of reciprocity, but only as far as do ut des, and he takes a more pessimistic view of their concern for justice. The characters of the Iliad, he explains, do not see the gods as dispensers of justice, rather “divinity is seen as an explanation for the tragic nature of life.”42 Kullman notes the conviction found among Iliadic heroes that the gods will punish wrongdoers, but also calls attention to the fact that the gods themselves can be the cause of any such wrongdoing.43 Like Lloyd-Jones, Kullman agrees that mortals are responsible for their actions, but asserts that errors made under the influence of the gods cause disproportionate suffering, rather than order and reciprocity.44 More recently, Yamagata has argued that although both mortals and immortals are concerned with morality, their ideas of what constitutes justice are separate. In Yamagata’s view, the gods are compelled by moira, and although they have the ability to accelerate or delay, they cannot change the ultimate outcome. Their justice is the perpetuation of moira, but this does not always align with the human concept of 42 Kullman (1985) 8. 43 44 Ibid. 15. 18 justice, and for this reason the gods appear to punish men beyond what is reasonable.45 Humans, on the other hand, attribute many moral functions to the gods, especially Zeus, but Yamagata finds little evidence that the immortals fulfil these roles.46 Allan, finally, is critical of those who label the gods of the Iliad as amoral. Not only does he claim that Zeus is the enforcer of cosmic justice, but also that he is “deeply concerned with the social norms of justice, both human and divine.”47 Like Lloyd-Jones, Allan comments that the system of justice among humans, which places the themistes in the hands of kings, is derived from Zeus himself. 45 Yamagata (1994) 97-120. 46 47 Allan (2006) 32. 48 48 Ibid., 9-10. 46 Ibid., 3-21. 46 Ibid., 3-21. 1. Iliad 21: Achilles and the Scamander Most critically, Allan asserts that morality is formed by social beliefs, and thus a person’s actions may be evaluated by the community and found to be either just or unjust according to those beliefs. Nevertheless, he maintains that such a fact does not exclude the gods from having an interest in human affairs.48 Overall the scholarship since Lloyd-Jones’ book has favoured his interpretation of the gods more than the amoral and immoral gods of Dodds and Adkins. Some, like Kullman, maintain reservations about the interest the gods have in justice on the mortal plane, but most agree that the gods are, at the very least, concerned with a wider, cosmic form of justice, even if it does not align with human principles. I, too, am inclined to agree with this line of thought. That the gods are invested in maintaining cosmic order is illustrated at the end of the mache parapotamios, when Hephaestus, under the direction of Hera, burns the entirety of the Scamander’s riverine environment. καίοντο πτελέαι τε καὶ ἰτέαι ἠδὲ µυρῖκαι, καίετο δὲ λωτός τε ἰδὲ θρύον ἠδὲ κύπειρον, τὰ περὶ καλὰ ῥέεθρα ἅλις ποταµοῖο πεφύκει: τείροντ᾽ ἐγχέλυές τε καὶ ἰχθύες οἳ κατὰ δίνας, οἳ κατὰ καλὰ ῥέεθρα κυβίστων ἔνθα καὶ ἔνθα πνοιῇ τειρόµενοι πολυµήτιος Ἡφαίστοιο (21.350-5). 19 The elms and willows burned, as did the tamarisks. The lotus also burned, and the reeds and the galingale. Everything that lived alongside the beautiful running river was set aflame. The eels suffered and the fish came out of the whirlpools, tumbling here and there in the fair stream, distressed by the fire of the clever Hephaestus. Mackie describes this event as a “brutal act perpetrated by higher powers,”49 and the effects of the fire are indeed widespread and severe. Yet they come at a point when they are much needed. The episode builds in intensity from the beginning of the book, and multiple violent acts are inflicted on both sides. Achilles kills and defiles a number of Trojan soldiers, his rampage fills the river with blood and corpses and obstructs its flow, and he attacks the river, engaging it directly in battle. In retaliation, the river rouses itself, floods the plains, and threatens to violate Achilles’ body beneath the riverbed (316-23). ( ) 52 The Scamander knows this to be the case: in its initial speech to Achilles the river does not ask him to desist, but, because Zeus has allowed the slaughter, requests he take the battle elsewhere (21.16-7); later the Scamander tells Hera that it will desist from the attack, accepting that Troy has been allotted an inexorable fate (373-6). The Scamander, like other gods, has the ability to delay the realisation of moira, but it cannot prevent the realization of moira indefinitely. πᾶν δ᾽ ἐξηράνθη πεδίον, σχέτο δ᾽ ἀγλαὸν ὕδωρ. 49 Mackie (2008) 184. ( ) 50 Redfield (1975) 251 n. 15. 51 ( ) 51 See Jones (2005) 23-4. 1. Iliad 21: Achilles and the Scamander As Redfield asserts: “as always in the Iliad, defilement brings about further defilement in a reciprocal process.”50 As soon as Hera and Hephaestus step in, the progression of violation halts. Not only do Hephaestus’ flames burn the corpses that were obstructing the river (343-4), thereby reversing the non- natural state of the water, they also weaken the river so it can no longer continue flooding (366). The fire also completes the funerary rites of the slain Trojan soldiers by cremating them.51 Once Hephaestus has withdrawn his flames, the river reverts to its usual, natural course (381-2). The restoration of the river also signals the end of delay and the continuation of the narrative. Furthermore, the intervention of the Olympian gods prevents the premature death of Achilles; he must live so that he can fight and kill Hector later, and thus accept his moira.52 Moreover, the simile used to describe the effects of the fire is unambiguously positive: 20 ὡς δ᾽ ὅτ᾽ ὀπωρινὸς Βορέης νεοαρδέ᾽ ἀλωὴν αἶψ᾽ ἀγξηράνῃ: χαίρει δέ µιν ὅς τις ἐθείρῃ (345-7). The whole plain was parched, and the shining water was held fast. Just as the north wind in late summer quickly dries a newly watered garden, and the one who cultivates it is glad. Once again domesticated nature is inserted into the narrative through simile. Despite the apparent brutality of the divine fire, the happiness of the farmer, indicated by the use of χαίρει, suggests that the ultimate outcome is positive: the fire in the river restores balance by reversing the non-natural phenomena and subduing the force that was inhibiting the trajectory of the narrative. In other words, the mache parapotamios is associated with a disruption in the cosmic order, in that it is a challenge to moira, and therefore a concern of the gods. However, there are potential concerns on the human plane also. Whether or not the gods are interested in justice among humans, there is, nonetheless, a set of norms for acceptable human behaviour. These include institutions such as oaths, xenia, piety, supplication, and treatment of the dead. The latter three are of particular importance to us, as the mache parapotamios episode sees them all challenged in some way. Although some Iliadic heroes escape divine punishment for impiety, overall the Greeks and Trojans are anxious to appease the gods and award them proper time. 53 Hence, at 16.249-52 Zeus grants only part of Achilles’ request: Patroclus will succeed in driving away the Trojans, but he will not return alive, compelling Achilles to rejoin the fighting and fulfill his moira. 1. Iliad 21: Achilles and the Scamander In book 1 Apollo unleashes a plague upon the Greeks; all involved, including Agamemnon and Achilles, realise that the only way to stop the plague is to return Chryseis to her father and sacrifice to Apollo (92-100; 442-5), thus reinstating his and his priest’s time. More generally, people express their piety through sacrifice (e.g. 2.400-3; 11.725-9) and offerings (e.g. 16.220-30; 6.297-310) in order to make a request, which may or may not be granted. Homeric religion works on the basis of reciprocal time, but because the gods are compelled to uphold moira they do not always appear to reciprocate in kind.53 Nevertheless, humans value their perceived relationship with the gods, perhaps because they believe the reciprocity works both ways, so the gods will reward piety and punish impiety. If we consider piety to be the 53 Hence, at 16.249-52 Zeus grants only part of Achilles’ request: Patroclus will succeed in driving away the Trojans, but he will not return alive, compelling Achilles to rejoin the fighting and fulfill his moira. 21 proper allotment of time to the gods, then it certainly seems to be the case that Achilles acts impiously in his treatment of the Scamander. Before his direct assault upon the river, the Greek hero scorns the Trojans for their pious behaviour: proper allotment of time to the gods, then it certainly seems to be the case that Achilles acts impiously in his treatment of the Scamander. Before his direct assault upon the river, the Greek hero scorns the Trojans for their pious behaviour: οὐδ᾽ ὑµῖν ποταµός περ ἐΰρροος ἀργυροδίνης ἀρκέσει, ᾧ δὴ δηθὰ πολέις ἱερεύετε ταύρους, ζωοὺς δ᾽ ἐν δίνῃσι καθίετε µώνυχας ἵππους (21.130-2). Not even your fair flowing, silver-eddying river will help you, to which for so long you have sacrificed many bulls, and hoofed horses which you cast, still alive, into the eddies.54 In fact, Achilles is not entirely correct; the river does rise up to help the Trojans, because the Scamander is domesticated, and because they have offered him appropriate time in the past. Achilles, who does nearly everything he can to disrespect the river, including insulting it, triggering non-natural phenomena, and directly attacking it, refuses to allot the river appropriate time and incurs its wrath. g p 55 He loses his footing almost immediately (21.241-2) then turns to flee (246-8), but i prevented from escaping the onslaught again and again until he prays to Zeus for aid (248 83). See Chaudhuri (2014) 201-3. 54 Despite Achilles’ scorn, he too has worshipped rivers: at 23.140-9 he cuts a lock of hair he had been reserving for the Spercheus river. 55 54 Despite Achilles’ scorn, he too has worshipped rivers: at 23.140-9 he cuts a lock of hair he had been reserving for the Spercheus river. 55 He loses his footing almost immediately (21.241-2) then turns to flee (246-8), but is prevented from escaping the onslaught again and again until he prays to Zeus for aid (248- 1. Iliad 21: Achilles and the Scamander 59 Pedrick, too, does not see any special code relating to supplication in the Iliad, and argues that the decision to accept or refuse is personal and without shame.60 Naiden, who maintains that supplication may be accepted or refused depending upon the crimes of the suppliant, evaluates the reasons given for rejection, noting that on the battlefield a suppliant may be rejected simply for his status as the enemy.61 Indeed, the gods do not punish any of the warriors who refuse their suppliants, nor do their peers shame them.62 Supplication in the Iliad has been the subject of much debate among critics, and any discussion of it requires careful consideration. Although many have noted the frequent failed supplications in the Iliad,56 it is also true that the narrative is framed by two successful supplications: Thetis to Zeus in book 1, and Priam to Achilles in book 24. Gould has argued that the rules of supplication are bendable; thus he suggests that the unsuccessful supplicatory episodes in Homeric narrative are not failed or rejected supplications, but incomplete supplications, which the supplicandus has every right to refuse because physical contact has not been made, or has been broken.57 Critics of this view note that his argument does not take into account context, or the importance of the verbal plea; battlefield supplication has an entirely different flavour to a supplication made in someone’s home in a time of peace.58 Kelly, who argues that battlefield supplication in the Iliad serves to characterise the supplicandi, does not believe that the refusal of the suppliant should in any way be considered immoral. 59 Pedrick, too, does not see any special code relating to supplication in the Iliad, and argues that the decision to accept or refuse is personal and without shame.60 Naiden, who maintains that supplication may be accepted or refused depending upon the crimes of the suppliant, evaluates the reasons given for rejection, noting that on the battlefield a suppliant may be rejected simply for his status as the enemy.61 Indeed, the gods do not punish any of the warriors who refuse their suppliants, nor do their peers shame them.62 Nevertheless, the very first supplication in the Iliad is a successful one; Thetis approaches Zeus on behalf of her son, grasps his knees and chin, and beseeches him to honour Achilles (1.500-16). Zeus bows his head, assenting to her request (528-30). 1. Iliad 21: Achilles and the Scamander Despite his lengthy boasting, the river proves too much for him, and Achilles barely manages to hold his own until the other gods step in to help.55 The mache parapotamios reinforces the superiority of the gods over mortals, even mortals such as Achilles, and also marks Achilles out as impious towards a specific god. However, Achilles’ impiety does not extend further than this one river deity; he is the favourite among many of the other gods, he makes due sacrifices, and his impiety here does not affect his relationship with the Olympians. The Achaeans, who presumably would not have worshipped the Scamander due to its geographical distance from Greece, would have viewed Achilles’ impiety at this moment as a non-issue. However, the Trojans, who worship the Scamander as they do other gods, would certainly have regarded Achilles’ actions as morally repugnant. This impiety, then, is localised; its occurrence does not violate a universal moral code and it is only immoral to some. 22 Supplication in the Iliad has been the subject of much debate among critics, and any discussion of it requires careful consideration. Although many have noted the frequent failed supplications in the Iliad,56 it is also true that the narrative is framed by two successful supplications: Thetis to Zeus in book 1, and Priam to Achilles in book 24. Gould has argued that the rules of supplication are bendable; thus he suggests that the unsuccessful supplicatory episodes in Homeric narrative are not failed or rejected supplications, but incomplete supplications, which the supplicandus has every right to refuse because physical contact has not been made, or has been broken.57 Critics of this view note that his argument does not take into account context, or the importance of the verbal plea; battlefield supplication has an entirely different flavour to a supplication made in someone’s home in a time of peace.58 Kelly, who argues that battlefield supplication in the Iliad serves to characterise the supplicandi, does not believe that the refusal of the suppliant should in any way be considered immoral. ) pp ( pp ) 63 For the characteristic gestures of supplication, see Gould (1973) 75-7. ( ) 61 Naiden (2006) 136. E.g. Diomedes refuses to accept Dolon as his suppliant because he may return later and do more harm to the Greeks (10. 449-53). 59 Kelly (2014) 166; Kelly suggests that modern critics, who value mercy as a social norm, find it difficult to look past the violence of the refused supplication, and thus have taken an anachronistic approach to the issue. 60 ( ) 62 Diomedes (10.454-7), Agamemnon (6.63-6; 11.143-7) and Achilles again (20.463-72 22.337-43) all refuse their suppliants (or the suppliants of others) and receive no punishment. ( ) 58 Naiden (2006) 11; Kelly (2014) 150. ( ) 57 Gould (1973) 81-2. 58 58 Naiden (2006) 11; Kelly (2014) 150. 59 ( ) 57 Gould (1973) 81-2. 58 pp 60 Pedrick (1982) 129. 56 Gould (1973) 80; Pedrick (1982) 132; Yamagata (1994) 41; Naiden (2006) 135; Kelly (2014) 147. 57 ( ) 57 Gould (1973) 81-2. 58 pp 60 Pedrick (1982) 129. 61 56 Gould (1973) 80; Pedrick (1982) 132; Yamagata (1994) 41; Naiden (2006) 135; Kelly (2014) 147. 57 Gould (1973) 81-2. 58 Naiden (2006) 11; Kelly (2014) 150. 59 Kelly (2014) 166; Kelly suggests that modern critics, who value mercy as a social norm, find it difficult to look past the violence of the refused supplication, and thus have taken an anachronistic approach to the issue. 60 Pedrick (1982) 129. 61 Naiden (2006) 136. E.g. Diomedes refuses to accept Dolon as his suppliant because he may return later and do more harm to the Greeks (10. 449-53). 62 Diomedes (10.454-7), Agamemnon (6.63-6; 11.143-7) and Achilles again (20.463-72; 22.337-43) all refuse their suppliants (or the suppliants of others) and receive no punishment. 63 For the characteristic gestures of supplication, see Gould (1973) 75-7. 1. Iliad 21: Achilles and the Scamander This is a textbook supplication: the suppliant physically lowers herself, performs the customary gestures, and makes her case persuasively.63 The supplicandus, for his part, 6 Gould (1973) 80; Pedrick (1982) 132; Yamagata (1994) 41; Naiden (2006) 135; Kelly 2014) 147. 7 23 hears her request and grants it, despite the trouble it will cause him. This episode sets up a paradigmatic example of supplication, which will not be seen again until Priam supplicates Achilles in the final book. The divine example establishes supplication as a norm; although there is no universal code that dictates the proper response to a suppliant, the refusal to accept perverts an established paradigm. The supplicandi of the Iliadic battlefield may not be immoral, but that does not mean their actions are any less distasteful. Achilles, who rejects the plea of Lycaon, incurs only the anger of the Scamander. Yet his actions, justified they may be, are liminal; the slaughter of a suppliant invites more pathos than the slaughter of a better-matched warrior such as Asteropaeus.64 A similar idea is at play when it comes to the treatment of the dead. Overall, soldiers on both sides express horror at the thought of mutilation by the enemy, or by scavenging animals such as dogs and birds. So Agamemnon threatens his soldiers with the prospect of corpse mutilation by dogs and birds (2.391-3) and Athena expresses her anger by suggesting that the Trojans will feed their fat and flesh to animals (8.379-80). Furthermore, Priam predicts his own grisly fate, to be devoured raw by his own dogs, which he describes as the most pitiable fate possible (22.66-76) and Hector begs Achilles in his last moments to spare him the dishonour of having his corpse mutilated by dogs (22.338-9). These are only a selection of the many instances in which characters express deeply negative feelings towards corpse mutilation, especially by animals. However, for the most part, animal corpse mutilation is reserved for threats and fears, and rarely does it directly feature in the narrative. It is, without a doubt, an undesirable fate to be consumed by animals, yet those who acknowledge its horror tend to accept it as par for the course in war, rather than condemn those who would cause it to occur. In book 21, fears of mutilation become real when fish and eels consume the corpses of Lycaon (122-9) and Asteropaeus (203- 4). 64 If nothing else, supplication is at least a narrative device employed to heighten the pathos of a moment; after all, Homer made the choice to include six battlefield supplications in his poem, when he needed not include any. As Segal (1971) 13 explains, “The audience, in order to experience the movement toward climax and resolution, must be emotionally involved in the events. The stakes must be something that matter.” 1. Iliad 21: Achilles and the Scamander The descriptions are grisly: fish and eels tear and pull at fat from the body. Segal calls this episode “a massive enlargement of the mutilation theme,” in which all Trojan corpses in the river are implicitly fed on. The outrage of the Scamander, according to Segal, is a result not only of Achilles’ general slaughter of Trojans, but 24 of his treatment of their corpses, too.65 In reality, the Scamander makes no specific comment to this effect. Moreover, the other gods do not express anger at Achilles for his treatment of the Trojans. While it is certainly the case that mutilation by animals is an undesirable fate, it is nevertheless not immoral to let an enemy be devoured by dogs, or any other animal. Even more direct mutilation, such as the treatment of Hector’s body at the hands of Achilles and the other Greeks, is also not necessarily immoral; in fact, the Greek soldiers do not hesitate to stab his lifeless body where it falls (22.369-71). While Apollo and Zeus do indeed denounce the brutal treatment of Hector’s body, they both refer to his unwavering piety as the reason for their pity (23.33-8; 66-70); Hector has always allotted proper time to the gods, and they in turn reciprocate, not by sparing his life, but by allowing his body to receive a proper burial. Although Achilles has clearly transgressed a norm by mutilating Hector’s body beyond what is, perhaps, reasonable, it does not mark him out as anything more than liminal. Furthermore, burial itself is an ideal, as illustrated by the cremation and funeral games for Patroclus in book 23, and the funeral of Hector in book 24. Yet these are isolated examples; it seems that the majority of warriors do not receive a proper burial or funeral.66 This is hardly surprising: the volume of bodies on the battlefield is too immense to allow every dead soldier a send-off. It is not immoral, then, to deny one’s opponent burial, nor is the lack of burial unexpected. Achilles is not the only hero to leave a trail of unburied corpses behind him, and though the piles of bodies in the river evoke an exceedingly vivid picture of slaughter, it is more indicative of Achilles’ ruthlessness and prowess as a warrior than any kind of moral degeneracy. g ( ) 68 Bespaloff’s words resonate here: “Who is good in the Iliad? Who is bad? Such distinction do not exist; there are only men suffering, warriors fighting, some winning, some losing,” (1947) 48. 66 Though there is a mass burial at 7.416-32. 67 65 Segal (1971) 31. 66 67 Yamagata (1994) 42. 68 67 Yamagata (1994) 42. 68 67 Yamagata (1994) 42. Segal (1971) 31. 66 Though there is a mass burial at 7.416-32. 67 69 Compared Thersites’ failures to Sarpedon’s statement that men must earn the respect of their peers by proving their strength and courage in battle (12.310-28). 70 p y p g g g ( ) 70 Taplin (1980) 15-6. Scully (2003) challenges Taplin’s reading, suggesting that the images on the shield do not present a place to which heroes wish to return, but a frightening vision of an unrecognisable world. 1. Iliad 21: Achilles and the Scamander That there is no strong, universal code of ethics in the Iliad should not come as a surprise; “in battle, killing is the business,” notes Yamagata.67 Indeed, while the Odyssey is quick to vilify those who transgress social norms, the Iliad rarely makes such judgements; there are no villains at Troy, only fallible people.68 The exception is perhaps Thersites: αἴσχιστος δὲ ἀνὴρ ὑπὸ Ἴλιον ἦλθεν (“the most shameful man to 25 have come to Troy”, 2.216) who insults both Achilles and Agamemnon and is quickly punished by Odysseus with a blow to his back (265-6).69 Nevertheless, war itself is not presented as immoral; it is only the other side to a coin, on the reverse of which is peace, or the cessation of conflict. This is Taplin’s ultimate argument, according to which the Iliad presents war in relation to peace, particularly on the shield of Achilles, and although the images of peace are idealised, neither it nor war is judged better than the other.70 Similar to Taplin, I propose that the images of domesticated nature on the shield and in similes represent peace as an ideal to aspire to, and for this reason they are distant and unambiguous. Undomesticated nature receives due mention also, often in its capacity to overcome domesticated nature, as war itself overcomes peace. Nevertheless, the triumph of war over peace and undomesticated nature over domesticated nature is neither negative nor positive; it is simply the reality of the Iliad. However, when nature becomes non-natural, as the Scamander does in Iliad 21, it is an aberration, an affront to the cosmic order, and must be dealt with. Indeed, although Hephaestus acts according to the will of Hera, whose only goal appears to be the preservation of Achilles and the destruction of Troy, he nevertheless succeeds in correcting the non-natural anomalies and restoring nature to its proper order. So too will Poseidon and Apollo restore the Trojan plain to its untouched state when they raze the rampart of the Greeks after the war has ended: δὴ τότε µητιόωντο Ποσειδάων καὶ Ἀπόλλων τεῖχος ἀµαλδῦναι ποταµῶν µένος εἰσαγαγόντες. ὅσσοι ἀπ᾽ Ἰδαίων ὀρέων ἅλαδὲ προρέουσιν, Ῥῆσός θ᾽ Ἑπτάπορός τε Κάρησός τε Ῥοδίος τε Γρήνικός τε καὶ Αἴσηπος δῖός τε Σκάµανδρος καὶ Σιµόεις, ὅθι πολλὰ βοάγρια καὶ τρυφάλειαι κάππεσον ἐν κονίῃισι καὶ ἡµιθέων γένος ἀνδρῶν (12.17-23). 1. Iliad 21: Achilles and the Scamander 26 Then Poseidon and Apollo deliberated about destroying the wall, leading towards it the might of the rivers, those which flow from Mount Ida to the sea: the Rhesus, Heptaporus, Caresus, Rhodius, Granicus, Aesopus, as well as the sacred Scamander and Simois, where many shields and helmets had fallen into the sand, and a race of semi-divine men. λεῖα δ᾽ ἐποίησεν παρ᾽ ἀγάρροον Ἑλλήσποντον, αὖτις δ᾽ ἠϊόνα µεγάλην ψαµάθοισι κάλυψεν, τεῖχος ἀµαλδύνας: ποταµοὺς δ᾽ ἔτρεψε νέεσθαι κὰρ ῥόον, ᾗ περ πρόσθεν ἵεν καλλίρροον ὕδωρ (30-3). λεῖα δ᾽ ἐποίησεν παρ᾽ ἀγάρροον Ἑλλήσποντον, αὖτις δ᾽ ἠϊόνα µεγάλην ψαµάθοισι κάλυψεν, τεῖχος ἀµαλδύνας: ποταµοὺς δ᾽ ἔτρεψε νέεσθαι κὰρ ῥόον, ᾗ περ πρόσθεν ἵεν καλλίρροον ὕδωρ (30-3). λεῖα δ᾽ ἐποίησεν παρ᾽ ἀγάρροον Ἑλλήσποντον, αὖτις δ᾽ ἠϊόνα µεγάλην ψαµάθοισι κάλυψεν, τεῖχος ἀµαλδύνας: ποταµοὺς δ᾽ ἔτρεψε νέεσθαι κὰρ ῥόον, ᾗ περ πρόσθεν ἵεν καλλίρροον ὕδωρ (30-3). He [Poseidon] calmed the current of the Hellespont, and when the wall was destroyed he covered the great beach with sand. Then he returned the stream to its course, where the fair-flowing water had run before. This wall was θεῶν δ᾽ ἀέκητι τέτυκτο \| ἀθανάτων (“built contrary to the will of the immortal gods,” 12.8-9), and although the gods destroy it for this reason, the razing of the wall has the added effect of restoring the environment to its original state. Furthermore, this glimpse into the future reminds us that even though the war will come to an end, the landscape will remain constant long after humans have left. It is clear that the Iliad presents the workings of nature as part of the cosmos, and the violation of nature therefore disturbs the cosmic order. Nature is also valued according to its relationship with humankind; therefore, there is an underlying need and desire to domesticate nature, just as there is a desire to end war and return to peace. But the near-defeat of Achilles in the face of the Scamander’s onslaught demonstrates that humankind’s struggle with undomesticated nature is not so easy to overcome, and that nature is a divine, eternal force that may be tamed and altered, but ultimately exists, and will continue to exist, beyond the works and activities of humankind. 27 2. Punica 4: Scipio and the Trebia The Punica of Silius Italicus is the longest extant Latin poem, with seventeen books and more than twelve thousand lines of verse. The Second Punic War is the subject of the poem, and the narrative follows the campaign of Hannibal as he enters Italy and makes his way towards Rome. There is no single Roman hero who occupies the focus of the epic; many fight and die for their city, including the celebrated Scipio Africanus, who sees out the end of the war. The narrative ends with the battle of Zama, in which Hannibal’s army is finally and decisively defeated. The mache parapotamios occurs in book 4 during the battle of the Trebia, an account of which is given below. Other significant battles that occurred near water will be referred to throughout this chapter, including the battles at the river Ticinus, Lake Trasimene, and Cannae (Aufidus river), as well as the incident involving Regulus and the serpent at Bagrada. The mache parapotamios begins when Hannibal drives the Roman soldiers towards the banks of the Trebia and into the water (4.570-2). At this moment the Trebia swells to attack the Romans, apparently roused by Juno’s prayers (573-4). Elephants join the carnage in the river (598-9) and drive the frightened Trebia before them (601-2). Then Scipio (father of Scipio Africanus) makes an appearance and wreaks such slaughter that the Trebia fills with the bodies of the slain: corporibus clipeisque simul galeisque cadentum \| contegitur Trebia, et vix cernere linquitur undas (“the Trebia was covered with corpses, as well as the shields and helmets of the fallen soldiers, and scarcely was it possible to see the waves,” 625-6). When the Trebia begins a fresh attack, Scipio addresses the river and rebukes it for betraying the Romans and allying itself with the Carthaginians (642-8). The river engages Scipio directly in a fight before rising out of the water to address him and complain about the bodies and blood in his stream (660-66). Scipio then beseeches the gods for help (670-5); Vulcan, under the command of Venus, sends his fires to beat the river back (675-89). The river is quelled and immediately afterwards the Po appears, apparently 28 also affected by the fires (690-1).1 Eventually, the river is restored to its proper course (“ripas… priores”, 696-7). It should be evident that Silius closely parallels Homer in his rendering of the mache parapotamios. ( ) p ( ) 4 Polybius’ version of the battle at the Trebia also notes that the river was in flood but says little else about the river. Nicol (1936) 32 suggests Appian as another possible source, who included Scipio in his account of the battle (The Hannibalic War 6-7), unlike Livy who claimed he was injured at the time (21.35-9). See Pomeroy (2010) 27-45 for a more recent discussion of Silius’ sources. g y 2 For a comprehensive analysis of the similarities and differences between the river battles o the two poems, see Juhnke (1970) 13-24. 1 The Po is an interesting addition to this episode, as it is referred to frequently throughout the poem as a victim of non-natural phenomena (1.131-2; 5.127-9; 9.187-8; 12.696-7), though no battles are fought in or near it. The Trebia, however, is a tributary of the Po. 2 Gossage (1969) 77; Miniconi-Devallet (1979) lxviii; Vessey (1982) 591; Feeney (1991) 302; Wilson (1993) 218. p , ( ) 3 Nicol (1936) 31; Spaltentstein (1986) 319. p , ( ) 3 Nicol (1936) 31; Spaltentstein (1986) 319. 4 2. Punica 4: Scipio and the Trebia Like the Scamander, the Trebia is allied with a specific group and directly attacks another. Moreover, the Trebia is filled with corpses, blood, animals and equipment, and it appears in anthropomorphic form to complain about this. The battle between the river and hero is inconclusive, and, as in Homer, the fires of Vulcan ultimately bring about the end of the episode. Although the episode in the Punica is much more condensed than that of the Iliad, it draws many of its major elements from Homer’s version.2 But Silius’ account of the battle has not been entirely lifted from the archaic Greek poet. The battle of the Trebia was a recorded historical event, described by Livy in book 21 of his Ab Urbe Condita. Though Silius certainly embellishes, some, such as Nicol and Spaltenstein, have noted that the river’s involvement recalls Livy’s statement that the Trebia was higher than usual at the time of the battle (21.54.9).3 Silius almost certainly used other sources, too, perhaps Polybius and Appian, who both also describe the battle of the Trebia.4 In any case, the narrative draws from an historical tradition; it selects its general plot and characters from attested historical events. Silius is not the first to write an historical epic, but his is one of the only extant poems that combines history with mythic material, a decision for which he has been much criticised.5 Indeed, the battle of the Trebia is one of the most prominent examples of this; Silius takes an historical battle and adds the personification of nature, superhuman strength, and divine intervention. Although the outcome of the battle reflects the historical record, the details are more reminiscent of Homer, Virgil and Ovid than Livy. 29 With that in mind, I will now outline instances of rivers in earlier Roman epic. Virgil’s Aeneid makes an obvious starting point.6 This text, although lacking a mache parapotamios, includes an episode in which the Tiber is personified. Aeneas, dozing beside the river, is visited by the river-god Tiberinus, who predicts the foundation of Alba Longa and advises Aeneas to visit Evander at Pallanteum (8.31-65). The river and river-god depicted here are tranquil and beneficent, vastly different to the Trebia of the Punica. 6 As is the case with most of the post-Virgilian poets, Silius owes much to Virgil. See, e.g.: Hardie (1993) and Pomeroy (2000). It is said that Silius venerated Virgil so much that he purchased the site of his tomb (Martial 7.63; 11.48). For scholarship on nature in the Aeneid see Benario (1978) and Thomas (1988). 7 ( ) ( ) 7 Jones (2005) 29-30. ( ) 7 Jones (2005) 29-30. 2. Punica 4: Scipio and the Trebia Even when the river halts the flow of its current, as it does later in order to carry Aeneas and his men to Evander (86-90), it does so at its own behest, not as a non-natural response to corpses in its stream. However, despite this picturesque image of the Tiber, the Sibyl makes clear the suffering it will experience as a result of war, and compares the river directly to the rivers of the Troad: bella, horrida bella, et Thybrim multo spumantem sanguine cerno. I see wars, terrible wars, and the Tiber, frothing with thick blood. Neither the Simois, nor the Xanthus, nor the Doric camp will be absent. Indeed, Latinus corroborates this image: recalent nostro Thybrina fluenta \| sanguine (“the Tiber’s stream still runs warm with our blood,” 12.35-6). The bloodying of the Tiber in the Aeneid lacks the same moralising force that similar events have in the Punica, as we will soon discover; nevertheless, the comparison between the Tiber and the rivers of the Troad implies that the Roman river will act similarly to the Scamander and Simois, which had supported the indigenous population, rather than the invading force. There are clear challenges ahead for the Trojan settlers and their relationship with a new environment.7 Although it is clear that Homer is the source model for Silius’ mache parapotamios, Virgil’s Aeneid still maintains a firm grip on 30 the overall arrangement of the episode: as von Albrecht points out, the present divine apparatus, and the introduction of unconcealed moral concerns, are more Virgilian than Homeric.8 Rivers are mentioned frequently in Lucan’s Civil War also. They have certain symbolic resonances; the Nile, in particular, acts as a boundary for the limits of human ambition, which Caesar attempts to overcome, without success, in his quest for imperial knowledge and power. In the Nile digression at 10.172-333, Caesar urges the priest to divulge the secrets of the Nile’s source. This information, however, is sacred and deliberately kept unknown. See Barrenechea (2010). 10 Cf. the shield of Hannibal 2.449-52. See Beneker (2011). 11 ( ) 11 Rivers in Lucan appear in other capacities too, such as the catalogue of rivers at 2.399-438. However, it is not my intention to conduct a full investigation into rivers and nature in the Civil War. See Zientek (2014) for such a discussion. ( ) 9 See Barrenechea (2010). 10 8 Von Albrecht (1964) 149. 9 Von Albrecht (1964) 149. 9 See Barrenechea (2010). 9 See Barrenechea (2010). 10 ( ) 9 See Barrenechea (2010). 2. Punica 4: Scipio and the Trebia Caesar’s desire to know the Nile’s source demonstrates his imperialistic ambition, which seeks to reach beyond human and environmental boundaries.9 The Rubicon is likewise important, acting as a physical boundary between Gaul and Italy, and a metaphysical boundary separating peace from war, and appropriate behaviour from inappropriate behaviour, just as the Ebro does in the Punica.10 Non-natural phenomena have a place in Lucan’s epic too: in a passage recalling the violent regime of Sulla, the corpses of those killed clog the Tiber until it slows to a trickle. Blood and gore fill its banks until it is forced to throw the bodies up onto the land in order to restore its current (Civil War 2.209-20). This is only one of many images used to describe the horrors at Rome, but the bloodied Tiber is saved for last; its destruction is the climax of Sulla’s proscriptions. As in the Punica, the bloody river is a clear sign that a transgression has occurred.11 Moreover, the Rubicon swells as Caesar makes to cross it (1.204-5), but Lucan is more scientific in his reasoning than Silius: tum vires praebebat hiems, atque auxerat undas (“then Winter granted it strength and increased its waves,” 217). The Rubicon is not personified in this episode, which is what we would expect to see in Silius, though hiems is partly personified and hinders Caesar’s advance with an air of intention. Lucan’s treatment of nature is thus, in a way, similar to Silius’; he utilises nature and its processes as a way of expressing moral judgements. It is evident that these two epic poets influenced Silius’ use of rivers in his own epic. Further influence likely came from Ovid’s Metamorphoses, Ennius, and 31 other genres.12 Nevertheless, Silius places clearer emphasis on the moral implications of destructive human interaction with rivers and other areas of nature than his predecessors, with the possible exception of Lucan.13 Nature has a major part to play in the Punica, primarily as an indicator of boundaries, a topic I will discuss in detail in due course. However, I am not so interested in the effectiveness, or non- effectiveness, of Silius’ epic style, nor in how he compares to his epic predecessors. Rather, I will evaluate his specific use and treatment of nature in the Punica, particularly during the mache parapotamios. 12 E.g.: Ovid: Metamorphoses 8.547-9.97, Theseus’ meeting with the personified river Acheloüs and Acheloüs’ fight with Hercules. Ennius: Annales 1.65; 66-9, in what is likely the tale of Romulus and Remus, the Tiber floods and flows backwards; Jupiter restores the Tiber to its proper course. Non-epic works: perhaps Statius’ Silvae 4.67-100 the Vulturnus river welcomes the rule of Domitian. 13 Although Silius, unlike Lucan and Virgil, never allows the Tiber to be sullied with blood and gore, Hannibal expresses interest in conquering it, insofar as it represents the city of Rome: hic labor Ausoniam et dabit hic in vincula Thybrim (3.511). However, like the Capitoline, the Tiber is beyond Hannibal’s limits. 5 The Volturnus wind and the goddess of Italy (see above) are the only other two examples. Also of note is Juno’s visit to Hannibal in the form of Lake Trasimene at 4.725-6. p y 14 E.g. the Rhone is described as territus (3.483); Lake Trasimene avoids making contact with a corpse (5.330-1); at Bagrada the river and its banks mourn the death of the sacred serpent (6.284-5); the semi-anthropomorphic Volturnus wind interferes in a battle (9.495-523); the goddess of Italy speaks out angrily against the Carthaginians and visits Claudius in the Roman camp (15.522-43). Roman camp (15.522-43). 15 The Volturnus wind and the goddess of Italy (see above) are the only other two examples. Also of note is Juno’s visit to Hannibal in the form of Lake Trasimene at 4.725-6. 2. Punica 4: Scipio and the Trebia As I have done for the Iliad, I will consider nature in its domesticated, undomesticated and non-natural aspects, with emphasis on the Trebia. Unlike the Iliad, nature features prominently in the Punica. Rivers, lakes, forests, mountains, plains, and the sea are important indicators of topography. More than this, the humans of the narrative directly interact with nature and must factor it into their activities. Hannibal must cross the Alps and lead his army over dangerous rivers. The Romans struggle with the Carthaginians among trees (5.480-516), and many drown in the rivers of Italy. Every major battle in the epic is fought adjacent to water, except the final battle at Zama. Weather is also significant, and Jupiter occasionally stirs up wind, rain and thunder above battlegrounds (e.g. 5.70-2; 12.609- 11). The Trebia is not, by any means, the only instance of personified nature in the Punica,14 but it is the longest (59 lines at least, more if one includes its involvement in the battle prior to Scipio’s entrance). It is also certainly one of the most significant personifications, and one of the few times that nature is anthropomorphised.15 The river lifts its head out of the water and speaks: tum madidos crines et glauca fronde 32 revinctum \| attollit cum voce caput (4.659-60), although up until this point it has fought Scipio in its ‘elemental’ form and seems only to appear anthropomorphised to address Scipio.16 Nevertheless, the river is personified throughout the entire episode in other ways; it rouses its waters at the behest of Juno (573-4), and fears (timentem) the advancing of the elephants (601). Active verbs also emphasise its intention: intumuit (638), sustulit (638), propellit (639), torquet (640), impulit (650), and pressit (650), among others. Because the Trebia is personified, it has volition. Depending on its actions, we are able to distinguish between its domesticated and undomesticated state; as long as the river acts according to its will, then it remains within these two categories. When the river performs according to Juno’s request and targets the Roman soldiers, rather than the Carthaginians, it shows it is obedient to a particular god, and therefore domesticated in that capacity. On the other hand, to the Romans, and especially to Scipio, the Trebia is an undomesticated river that poses a threat to Roman safety. 16 As the Tiber does for Aeneas at Aeneid 8.31-5. 2. Punica 4: Scipio and the Trebia This is clear in Scipio’s words to the Trebia: magnas, o Trebia, et meritas mihi, perfide, poenas exsolves, inquit, lacerum per Gallica rivis dispergam rura atque amnis tibi nomina demam; quoque aperis te fonte, premam, nec tangere ripas illabique Pado dabitur. quaenam ista repente Sidonium, infelix, rabies te reddidit amnem? (643-8) Treacherous Trebia, you will suffer punishments as great as you deserve. I will scatter the separated channels of your river through the land of the Gauls and I will deprive you the name of river. More than this, I will halt you at the spring from where you appear. And you shall not touch the banks, nor will you be allowed to be channeled into the Po. What sudden insanity, wretched Trebia, has turned you into a Carthaginian river? 33 Scipio’s speech indicates his belief that the Trebia is sympathetic to the Carthaginians, and, more pointedly, that it is not sympathetic to the Romans, though it should be, being an Italian river. However, there is something more sinister underlying Scipio’s threats. François Spaltenstein asserts: “‘Nomina’ symbolise l’existence. L’idée n’est pas que ce “fleuve” sera reduit, par example, à un “ruisselet”, mais que la Trébie disparaître.”17 So what exactly is Scipio suggesting here? At first, he seems to insinuate that he will punish the river by domesticating it, by forcing it to flow in separate channels throughout Gallic lands. This would be non-natural domestication, such as we saw in the Odyssey with Circe’s forced domestication of men who stray into her path.18 However, Scipio goes on to say that he will literally remove the river from the landscape; he will stop it at its source. This is not domestication as we have thus far seen it; this is complete annihilation. Domestication requires the existence of the river’s phusis, even if it is thwarted, but a river that does not exist has no phusis at all. Indeed, following Scipio’s threats, the Trebia responds: poenasne superbas \| insuper et nomen Trebiae delere minaris, \| o regnis inimice meis? (“Insolent man and enemy of my kingdom, do you threaten to punish me further by obliterating the name of Trebia?” 660-2). The river’s words confirm Scipio’s intention: he threatens to wipe the Trebia from the landscape altogether I will return to the substance of Scipio’s speech later, but for now I will consider the non-natural phenomena that affect the river. 18 See chapter 1. 19 The image of the river running backwards occurs, perhaps most famously, in Euripides’ Medea (410-20) to suggest a crisis in the social order. We also see this in Ennius (Annales 1.65). 17 Spaltenstein (1986) 319-20. 18 18 See chapter 1. 19 17 Spaltenstein (1986) 319-20. 18 2. Punica 4: Scipio and the Trebia Many of these are similar to what we have seen in the Iliad: the Trebia is filled with blood (593), corpses and equipment (625-6; 662-4) and it is unable to flow in its proper channel due to the obstructions (664). However, Silius adds an extra horror: the river is not only stuck, its stream also flows backwards (retroque feruntur) in its bed (665).19 When the river addresses Scipio, it rebukes him for the carnage he has wrought in the water and complains about each of the non-natural phenomena. Similar to the Scamander at Iliad 21.217, it suggests that Scipio either exercise moderation, or take his slaughter elsewhere: adde modum dextrae aut campis incumbe propinquis (666). Its words clearly suggest a concern for the preservation of its phusis. 34 In fact, all of these non-natural phenomena are thematised throughout the epic, and references to such events are common. Very early in book one, Juno predicts the future Roman defeats: dum Romana tuae, Ticine, cadavera ripae non capiant Simoisque mihi per Celtica rura sanguine Pergameo Trebia et stipantibus armis corporisbusque virum retro fluat ac sua largo stagna reformidet Thrasymennus turbida tabo, dum Cannas, tumulum Hesperiae, campumque cruore Ausonio mersum sublimis Iapyga cernam teque vadi dubium coeuntibus, Aufide, ripis per clipeos galeasque virum caesosque per artus vix iter Hadriaci rumpentem ad litora ponti (1.45-54). When your banks, Ticinus, cannot hold the bodies of the Romans, and the Trebia, my own Simois, flows backwards through the Gallic land, packed with Roman blood and the arms and bodies of men, and Lake Trasimene fears its own muddy pools, overflowing with putrid fluids. When I see Cannae from above, the grave of Italy, and the fields of Iapygia immersed in streams of Roman gore, while you, Aufidus, uncertain of your course as your banks come together, scarcely forge a narrow passage to the shore of the Adriatic Sea through the round shields and helmets and hewn limbs of men. In this programmatic secondary proem,20 the goddess refers to each Roman defeat not in terms of the actual battles, nor number of lives lost, but in terms of the ecological crises that occur in the wake of Carthaginian victory. Hannibal and his allies make similar boasts at 6.706-8, 11.185-9 and 135-41. 20 Recalling Jupiter’s prediction at Aeneid 1.257-96. 20 Recalling Jupiter’s prediction at Aeneid 1.257-96. ( ) 22 Indeed, the major places where nature is violated are in Italy: Ticinus, Trebia, Trasimene, Aufidus (all of which are included in Juno’s speech), and the Metaurus. The battle of Zama features no such violation. Other places outside of Italy are violated, such as the Rhone and the Druentia, as well as the Pyrenees and the Alps, but these are not inflicted with non-natural phenomena as the rivers of Italy are. p y 23 In fact, Livy recorded the earthquake described here (22.5), claiming that none of the combatants felt it, despite its tremendous strength. He does not include the detail that the Underworld was revealed through a crack in the ground, but he does say that the quake was strong enough to derail streams from their channels, force the sea into rivers, and cause major landslides. 21 Santini (1991) 69-72. 22 21 Santini (1991) 69-72. 22 2. Punica 4: Scipio and the Trebia Silius encourages his audience to view the Carthaginian enemy as a threat to the physical landscape of Italy; he undermines any glory that might be found in their victories by presenting them as transgressions against nature. In a detailed analysis of this passage, Carlo Santini 35 offers a number of interpretations based on Juno’s references to the violation of the rivers and lake in question. In particular, he proposes that the destruction of nature suggests an imbalance in the order of nature, as well as demonstrating that violence towards nature causes pollution, which is associated with religious taboos. He also proposes that the violation of nature works towards a climactic obliteration of nature.21 Underlying this argument is the certainty that the violation of nature, as seen in non-natural phenomena, is depicted negatively. More importantly, this passage indicates that the destruction of nature in the Punica is a direct threat to the safety and security of Italy and Rome.22 The idea that violent interference causes imbalance in nature is evidenced elsewhere in the epic also. The most striking example of this occurs during the battle at Lake Trasimene: as Hannibal charges into combat against Flaminius, the earth shakes open to reveal the Underworld below, the lake rises higher than it has ever done before to immerse the surrounding forests in water, and rivers and ocean currents flow in reverse (5.611-26).23 Such a disturbing event highlights the excessive climax of the battle; the effects of the fight literally cause the boundaries between planes to shift, and environmental phenomena contravene physical laws. Furthermore, the image of the river in reverse appears elsewhere to signal distressed nature: the Rhone runs backwards in fear of Carthaginian elephants at 3.463-5; the Ticinus, frightened of Mars, also reverses its course (4.442-3); and the Trebia complains that the volume of bodies in its stream forces it to run backwards (4.665). Beyond watercourses, the disruption of the cosmic order is also evidenced in other parts of nature: at 8.641-55 the Romans dream apocalyptic visions of blood flowing from Jupiter’s temple, the Allia rising above its banks, the Alps and Apennines shifting positions, meteor showers, and the breaking open of the heavens to reveal Jupiter himself. 24 Spaltenstein (1986) 321 notes: “L’encombrement du fleuve (vers 664) est devenu ensuite un motif constant, comme aussi ‘retro…feruntur’ 665.” , 25 This passage recalls the moment when Aeneas first catches sight of the Tiber, spilling forth its water into the sea while birds above sing and flit through the trees at Aeneid 7.30-4. 24 Spaltenstein (1986) 321 notes: “L’encombrement du fleuve (vers 664) est devenu ensuite 24 Spaltenstein (1986) 321 notes: “L’encombrement du fleuve (vers 664) est devenu ensuite 2. Punica 4: Scipio and the Trebia All of these dramatic events occur only in dreams, but they suggest that the 36 oncoming battle at Cannae will not be a favourable one; indeed, in the following passage a soldier predicts the Roman defeat, concluding: pons ecce cadentum \| corporibus struitur, reicitque cadavera fumans \| Aufidus (“See how the bodies of the fallen make a bridge, and the stinking Aufidus flings the corpses out of its stream,” 668-70). The images of nature in turmoil are horrific; they clearly suggest that something is amiss. The previous quote also illustrates that there remains a close causal relationship between violence towards nature and pollution.24 The rivers where the major battles are fought are all inundated with blood, bodies, and equipment, causing the water to become polluted. At Ticinus, the horror of the violence is emphasised by the description of its usual behaviour just prior to the battle: caeruleas Ticinus aquas et stagna vadoso perspicuus servat turbari nescia fundo ac nitidum viridi lente trahit amne liquorem. vix credas labi: ripis tam mitis opacis argutos inter volucrum certamine cantus somniferam ducit lucenti gurgite lympham (4.82-7). The translucent Ticinus keeps its waters clear and prevents its pools from being muddied in the shallow bed. Slowly it pulls its water along the stream, which shines a bright green. Scarcely would you believe it to be moving: so placidly along the shaded banks it travels, the birds singing melodiously in contest with one another. Thus it leads its soporific waters in shining eddies. Silius’ Ticinus is a peaceful, slow-flowing river,25 its waters clear and blue. Yet the river after the battle could hardly be more changed: Serranus later laments that the Ticinus is ater \| stragibus (“black with carnage,” 6.107-8), just as Juno had predicted 37 it would be overwhelmed with corpses at 1.45-6, a far cry from the crystal-clear stream it used to be. As Littlewood notes, the motif of the corpse-filled river emphasises the pollution of Italian land.26 In fact, the theme of pollution extends throughout the epic and is frequently used to vilify the Carthaginians. ( ) 27 See, for example, Hannibal’s dream of a huge serpent that crushes forests and destroys everything in its path (3.185-197); so will Hannibal crush Italy with war. Hannibal receives this omen with joy (215-6). 26 Littlewood (2011) 185. 27 26 Littlewood (2011) 185. 27 2. Punica 4: Scipio and the Trebia Hannibal’s boasts are echoed by Roman lament (Serranus at 6.106-12) and divine disapproval (Jupiter at 12.695-7); the deeds for which Hannibal appears most proud are presented as extreme, sacrilegious and transgressive.27 Time and again, nature rebels against the abuse it receives during war: the Druentia drowns many of Hannibal’s men as they attempt crossing (3.468-76), Lake Trasimene recoils from the body of a dead soldier (5.330- 1), and the Aufidus spews out corpses which had fallen into its stream (10.319-20). Although the major form of pollution – blood and gore in the river – results from the slaughter of both Carthaginian and Roman soldiers, the blame (or credit) for this defilement is almost always placed on Hannibal. Indeed in book 15 the goddess of the Italian earth herself appears and bemoans the treatment she has received at the hands of Hannibal: quot corpora texi \| caesorum, stratis totiens deformis alumnis! (“how many bodies of the slaughtered have I buried? How often am I deformed by the scattering of my own children?” 15.530-1). She complains too that the crops are now harvested with swords, and that she is made ugly by the destruction of houses which fall in gremium (532-5). But her anger is directed solely at Hannibal and she seeks aid from a Roman consul, condemning the Carthaginians to a mass grave (547-57). It is clear from her perspective that the Carthaginians alone are the cause of her suffering. Yet while the Italian goddess blames Hannibal for her troubles, the anthropomorphic Trebia assigns all blame for the pollution and reversal of its waters to Scipio. Indeed, this is not unwarranted; the appearance of Scipio signals the beginning of a bloodbath in which the Trebia is packed so closely with bodies that the water is barely visible (4.625-6). This is at odds with later Roman recollection of the event: Serranus laments the bloodying of the Ticinus and the Po, but of the Trebia he states only tuque insignite tropaeis \| Sidoniis Trebia (“and you, Trebia, distinguished by Carthaginian victory,” 6.108-9), omitting any mention of Scipio’s responsibility 38 for the pollution of the river. Scipio’s actions in this instance are problematic, because Silius generally utilises non-natural events and their causation as a way to transfer blame for Roman suffering onto the Carthaginians. Yet here, the non-natural events are clearly linked with the arrival of Scipio. Tipping (2010a) 193 4. 29 A commentary and discussion of this passage can be found in Bassett (1955) 1-20. 30 28 Tipping (2010a) 193-4. 29 30 Spaltenstein (1986) 410. 29 A commentary and discussion of this passage can be found in Bassett (1955) 1-20. 30 Spaltenstein (1986) 410. 28 Tipping (2010a) 193-4. 29 y p g ( ) 30 Spaltenstein (1986) 410. Tipping (2010a) 193-4. 29 A commentary and discussion of this passage can be found in Bassett (1955) 1-20. Tipping (2010a) 193 4. 9 A commentary and discussion of this passage can be found in Bassett (1955) 1-20. 0 Spaltenstein (1986) 410. 2. Punica 4: Scipio and the Trebia In fact, this is not so problematic; like Livy, Silius has pairs of good and bad consuls – Fabius and Minucius, Paulus and Varro – but none are perfect models of Roman heroism and virtue. For some, such as Minucius and Varro, their flaw is an irresponsible brashness and thirst for battle; others, such as Fabius, are perhaps too cautious to make much headway for Rome.28 Scipio’s major flaw is his inability to control his furor in battle, and this comes to light during his fight with the Trebia. The Trebia episode parallels the tale of Regulus and the serpent of Bagrada in book 6.29 The elderly Marus tells the story of Regulus, a celebrated Roman hero of the First Punic War, who fought and killed a monstrous serpent at the river Bagrada (6.140-298). Following the death of the serpent, Marus claims that the river, trees, caves and banks joined in lamentation because the serpent had been the servant of the naiads (283-8; 288-90). Furthermore, Regulus’ defeat of the serpent is said to foreshadow later peril (seris…periclis, 290), likely hinting at the future capture of Regulus.30 The destruction of nature is here clearly connected with military failure; the death of the serpent is almost a causal factor in Regulus’ future defeat. It is curious then, that Serranus, who happens to be Regulus’ son, should interrupt Marus’ tale with this exclamation: huic si vita duci nostrum durasset in aevum, huic si vita duci nostrum durasset in aevum, non Trebia infaustas superasset sanguine ripas (296-7). non Trebia infaustas superasset sanguine ripas (296-7). If the life [of Regulus] had continued on into our age, the Trebia would not have overflowed its ill-omened banks with blood. Regulus’ defeat of the serpent is heroic, certainly, and proves his valour in battle. However, the episode concludes with a problematizing of Regulus’ treatment of 39 nature: he has killed a sacred creature, for which he will be punished.31 Serranus’ intention is surely to evoke the unquestionable heroism of his father and point out that, had he been alive still, he would have led the Romans to victory. His words imply that the Trebia would not have been filled with the bodies of soldiers had Regulus been present at the battle. Or, perhaps, that Regulus would have prevented the need for the battle altogether, having already won the war at an earlier stage. 32 Regulus’ heroism is especially questionable, as summarized by Cowan (2010) 341: “Regulus is a problematic figure, whose courage and constantia are tainted by his recklessness, arrogance, and cruelty to his family.” See also Augoustakis (2006), especially 157-60 for more on Regulus at Bagrada. 33 Excuses may be made for Scipio’s behavior: Chaudhuri (2014) 209 notes that the Trebia is somewhat insincere in its accusation of Scipio, as the river began to be obstructed by bodies before his arrival, and it directly assaulted the Romans. Nevertheless, Silius saves the explicit description of the obstructed river for the moment right after Scipio appears on the scene; Scipio is presented as the major cause of the build-up of corpses in the river. 34 31 Augoustakis (2006) 159: “Regulus’ deed is explicitly described as a violation agains nature”. p p j p p 34 Santini (1991) 83. A similar point may be made about Regulus: Augoustakis (2006) 160 asks: “His fight against the serpent… presents a picture of violation. In this respect, how different is Regulus from Hannibal and his transgression in crossing the Alps?” 2. Punica 4: Scipio and the Trebia Yet there remains an undercurrent of ambiguity; it was, after all, Scipio who was responsible for the majority of the carnage in the Trebia, and without his presence the Romans would surely have faced a much more severe defeat, at least within the confines of the Punica. Although this is little more than an inconsistency, it nevertheless draws attention to a parallel between Regulus and Scipio, as Roman heroes who fight and triumph over a foe from nature with ambiguous, even questionable, results.32 The Bagrada episode illustrates how Silius uses human interaction with nature as a way to offer moral judgements. Regulus’ actions are not enough to vilify him completely, but they do cast a shadow over his memory. Similarly, Scipio’s encounter with the Trebia paints him as a liminal figure in this instance.33 Indeed, Scipio’s disregard for nature resembles the Carthaginian attitude. As Santini observes: “the Roman hero’s role might just as easily have been taken by Hannibal, no less arrogant than he.” Scipio’s status as Roman consul and hero does not necessarily justify his behaviour.34 Furthermore, not only does Scipio defile the sacred dwelling place of a god, he also threatens it with the worst fate imaginable: complete obliteration. Scipio’s threat introduces a new way of interacting with nature that is not present in Homer. Achilles (who has little power against the river), can barely hold 40 his own against it, let alone find a way to destroy it entirely. Yet in the Punica, Scipio fares much better against the river than his Homeric counterpart. Chaudhuri observes that Scipio’s threats are technological; they resonate with ideas of Roman dominance over waterways and suggest advances in hydroengineering.35 However, Scipio’s aim is not to take control of the river and govern it with technology, but in fact, to destroy it entirely. This is a step further than domination, shifting the attitude beyond the realm of natural and non-natural, onto a plane where humans have total control over nature without concern for its phusis. The poem does not explicitly praise or condemn this specific idea, but in the katabasis of book 13, Scipio, now deceased, advises his son to control himself in battle (martis moderare furori), and to learn from his father’s mistakes (13.667-71). Marks notes that the advice given seems to specifically recall the Trebia episode, when Scipio showed himself capable of excessive furor. ( ) 37 Compare the catalogue of Hannibal’s virtues and vices in Livy 21.4. Here Hannibal is likewise liminal owing to his many vices, though he does not reject nature’s limitations as he does in the Punica. ( ) 36 Marks (2005) 140-1. 37 35 Chaudhuri (2014) 208. Chaudhuri (2014) 208. 36 Marks (2005) 140-1. 2. Punica 4: Scipio and the Trebia Moreover, the language Scipio uses to encourage restraint in his son recalls the Trebia’s words to Scipio during the battle: adde modum dextrae.36 Later, Scipio’s son meets Hannibal’s father, Hamilcar. Scipio Africanus rebukes Hamilcar for the suffering Hannibal has caused. In particular, he says: Alpes \| eluctatus adest, fervet gens Itala Marte \| barbarico, et refluunt obstructi stragibus amnes (“he [Hannibal] came to us after struggling over the Alps, and the Italian nation is ablaze with foreign war, while the rivers flow backwards, piled high with the slaughtered,” 13.741-3). Scipio Africanus frames the war in terms of ecological crises, as Juno had done in book 1, but to him they represent the very worst of Carthaginian achievements, a level to which he will not stoop, unlike his own father. While Scipio Africanus assumes the mantle of the quintessential Roman hero, Hannibal descends further into moral disgrace as the narrative progresses. From the beginning, Hannibal’s campaign is marked by his ability to transcend boundaries within nature; he refuses to sleep (1.245-6), passes unfazed beneath lightning storms (253-5), marches without tiring (255-6), refuses to drink even on the hottest of days (257-60), and is the first of his men to swim across unfamiliar rivers (263-4).37 The treaty between Rome and Carthage dictates that the Carthaginians may advance no further than the Ebro, a limitation that Hannibal scorns: foedera, faxo \| iam noscas, 41 quid vana queant et vester Hiberus (“now I will make you understand that your treaty is good for nought, and your Ebro too,” 1.479-80). Not only does Hannibal break the treaty by physically moving his army far beyond the Ebro, he symbolically asserts his contempt for the barrier by wielding a shield that bears this image: extrema clipei stagnabat Hiberus in ora, extrema clipei stagnabat Hiberus in ora, curvatis claudens ingentem flexibus orbem. Hannibal, abrupto transgressus foedere ripas, Poenorum populos Romana in bella vocabat (2.449-52) The Ebro flowed around the outermost edge of the shield, surrounding the huge circle with its winding curves. Hannibal was there, crossing over the banks and breaking the treaty, as he urged the Carthaginians to war against the Romans. The description of the shield is programmatic, drawing on Homer’s description of Achilles’ shield in Iliad 18 and Virgil’s description of Aeneas’ shield in Aeneid 8. y g g g 39 As the shield of Achilles is a “microcosm of society”. See Hardie (1986) 340-2. 40 38 Hesiod, Theogony 337-45. Notably, Oceanus is missing from Aeneas’ shield; Jones (2005) 74 argues that the omission of the oceanic border is deliberate and “shows how the Romans have moved beyond imagining the edges of the earth and now touch them with their empire.” 40 Vessey (1975) 404. 40 Vessey (1975) 404. es od, g y 7 . o b y, Oce us s ss g o e e s s e d; Jo es ( 74 argues that the omission of the oceanic border is deliberate and “shows how the Roman have moved beyond imagining the edges of the earth and now touch them with their empire.” have moved beyond imagining the edges of the earth and now touch them with their empire. 39 As the shield of Achilles is a “microcosm of societ ” See Hardie (1986) 340 2 8 Hesiod, Theogony 337-45. Notably, Oceanus is missing from Aeneas’ shield; Jones (2005) 74 th t th i i f th i b d i d lib t d “ h h th R ( ) ( ) 43 See Augoustakis (2003) 250. 42 Following his successful, but less remarkable, crossings of the Pyrenees (1.415-19), the Rhone (446-65) and the Druentia (466-76). 41 The Alps hold further significance as a prefiguration of the walls of Rome. See von Albrecht (1964) 24-29. 42 41 The Alps hold further significance as a prefiguration of the walls of Rome. See von Albrecht (1964) 24-29. 42 Following his successful, but less remarkable, crossings of the Pyrenees (1.415-19), the 2. Punica 4: Scipio and the Trebia This passage in particular is modelled on Achilles’ shield, which is encircled by Oceanus, the father of rivers and border of the world.38 In the Punica, the Ebro takes the place of Oceanus, transforming the Punic War into a microcosm.39 But this microcosm is narrow and limited, as Vessey has argued, and serves to demonstrate that the success of Hannibal is temporary, “bound within a world restricted by space and time.” By contrast, Rome’s legacy will continue, sine fine.40 Hannibal’s crossing of the Ebro is a transgression of a boundary that is both microcosmic and part of nature, and a suggestion that his future campaign is bound to fail in the end. Boundary-crossing is of particular importance to the Punica. The Ebro is both a physical boundary separating Carthage’s territories from Rome’s, and a metaphysical boundary between appropriate and inappropriate human behaviour. The crossing of such frontiers held political resonance: Caesar’s bridging of the Rhine, for example, symbolically expanded Rome’s empire and challenged the people who 42 dwelt on the other side (Gallic War 4.16-18), just as Hannibal’s crossing of the Ebro challenges the current political and martial status quo. Rivers make especially good physical boundaries, owing to their inherently divisive nature, but mountains, too, mark borders between lands. The Alps act as another major boundary in the Punica – more important, even, than the Ebro, whose crossing is not narrated in the epic – due to their largely inviolate state, having not been traversed since Hercules did so many years previously (Punica 3.496-9).41 By crossing the Alps, 42 Hannibal rejects their divine inscrutability, clearly transgressing the line between what is appropriate and inappropriate. Even the men are at first uneasy at such an idea: at miles dubio tardat vestigia gressu, 43 See Augoustakis (2003) 250. p Albrecht (1964) 24-29. 42 at miles dubio tardat vestigia gressu, impia ceu sacros in fines arma per orbem, impia ceu sacros in fines arma per orbem, natura prohibente, ferant divisque repugnent (3.500-3). But the soldiers advanced slowly, moving their feet with uncertainty, as if they carried ungodly arms across the world into an inviolate land, acting against both nature and the gods. Their fear is incomprehensible to Hannibal, who urges them on with the promise of further victories (504-13). The army makes it over the mountains, but Hannibal’s success does not come without a struggle. Nature rebels against his army: the snow swallows many of the soldiers and the wind harries them as they advance (3.520-7). Despite these obstacles, the crossing is a success. Once again, Hannibal proves himself capable of passing beyond nature’s boundaries, but his army does not leave the Alps without altering the landscape: mutatur iam forma locis: his sanguine multo \| infectae rubuere nives (“now the appearance of the place was changed: here the snow grew red with copious blood,” 547-8).43 The stain they leave on the Alps is a sinister 43 reminder of Hannibal’s transgression, which looks forward to later battles that see rivers and fields stained with blood as well.44 Hannibal’s disregard for nature is apparent elsewhere too. A lengthy digression describes a peaceful episode in which the legendary Falernus entertains Bacchus at his home on the slopes of Mount Massicus (7.162-211). The place in question is made fertile through the beneficence of the god, and becomes one of the most famous wine-producing regions thereafter. Immediately following this passage, Silius declares: haec tum vasta dabat terrisque infestus agebat \| Hannibal (“then Hannibal laid waste to this place, and violence broke into the land,” 212-3). 45 The immediacy of Hannibal’s assault, which is no more than an expression of frustration at Fabius’ continuing refusal to join battle (213-4), on a place that Silius unequivocally describes as being among the most fertile of areas, serves to emphasise Hannibal’s utter disregard for the landscape – a quality that confirms him as improbus. Throughout the epic, Hannibal and his allies frame their conquests in terms of their triumphs over nature. Hannibal’s brother says of him: mihi, cui cedunt montesque lacusque \| et campi atque amnes, frater (“he is my brother, to whom mountains, lakes, fields and rivers yield,” 15.748-9). 45 This passage closely parallels Ovid’s story of Baucis and Philemon in the Metamorphoses (8.612-727). However, Silius’ version ends with greater violence. On Silius’ use of Ovidian material, see Wilson (2004) and Bruère (1959). 44 The motif of the Alps has been discussed in detail by Subrt (1991) who critiques and builds on the argument of von Albrecht (1964). 4 , ( ) ( ) 46 Manolaraki (2010) 311. , ( ) 46 Manolaraki (2010) 311. at miles dubio tardat vestigia gressu, However, despite Hannibal’s frequent boasts that he has mastered nature, his ability to overcome nature’s boundaries and limitations declines as the narrative progresses. Manolaraki connects the tidal imagery of the Punica, in particular the passage describing the Atlantic tides at 3.45-61, with Hannibal’s fate, governed, as it is, by cosmic necessity. While Hannibal is witness to the flow of the tide over the land, Manolaraki argues that he is not aware of the ebb of the tide as it retreats towards the ocean; this vision is reserved for Silius and his audience alone. Hannibal, who sees only the rising of the water, remains unaware of its eventual withdrawal and, analogously, his impending failure: “the tidal motif enables Silius… to illustrate that Hannibal underestimates the natural, divine, and historical principles that govern his place in the world.” 46 Indeed, Hannibal’s ability to withstand nature’s compulsion dwindles in the second half of the 44 epic. Once he could ride unassailed beneath a thunderstorm (1.253-5), but later, when Jupiter unleashes a furious tempest above the Carthaginians in defence of Rome, Hannibal is twice forced to retreat (12.627-32; 661-3).47 Similarly, when Hannibal attempts to return to Italy after setting sail for Carthage, Neptune overwhelms his fleet with a fierce storm that nearly kills him (17.218-90). Hannibal’s mastery over nature diminishes as his failure looms; Jupiter had allowed him to go so far: Tyrrhenas sternere valles \| caedibus, et ripas fluviorum exire Latino \| sanguine fas fuerit (“it is permitted for you to scatter the Etruscan valleys with corpses, and to make the banks of the rivers flow with Roman blood,” 6.602-4), but the father of the gods imposes limits upon Hannibal too: Tarpeium accedere collem \| murisque aspirare veto (“but I forbid you to reach the Tarpeian hill and conquer the city walls,” 604-5).48 Hannibal, however, is unaware of these limits; his campaign is characterised by the enduring and arrogant belief that he is capable of anything. 49 The crossing of nature’s boundaries, in particular the Alps, and the destruction of the Italian landscape at places such as the Ticinus, Trebia, Po and Trasimene, serve, in Hannibal’s mind, as evidence of his ability to succeed, because he has already triumphed in ways no others have done before. To Silius and his audience, these trespasses indicate instead that Hannibal will never conquer Rome, as these actions are transgressions that will eventually lead to his downfall. 48 Jupiter’s limitations on Hannibal recall Zeus’ restrictions on Achilles and Patroclus in the Iliad: both will win glory, but neither will take the city walls because they are restrained by moira (Iliad 16.707-9). Turnus of the Aeneid is also a strong parallel for Hannibal; he too is bound by a particular fate (Aeneid 10.622-5). y p ( ) 49 On Hannibal’s improba virtus see Tipping (2010) 61-73. y p ( ) 49 On Hannibal’s improba virtus see Tipping (2010) 61-73. 47 On this leitmotif see Schrijvers (2006) 103. 48 moira (Iliad 16.707 9). Turnus of the Aeneid is also a strong parallel for Hannibal; he too i bound by a particular fate (Aeneid 10.622-5). 47 On this leitmotif see Schrijvers (2006) 103. 48 j ( ) 48 Jupiter’s limitations on Hannibal recall Zeus’ restrictions on Achilles and Patroclus in the at miles dubio tardat vestigia gressu, Indeed, during the final battle at Zama, Silius narrates the deaths of a multitude of renowned Carthaginians at the hands of Scipio Africanus; these men, says Silius, were famous for their exploits at Saguntum, for polluting Trasimene and the Po, for marching against the dwelling place of Jupiter on the Capitol, and for profaning the sacred spaces of the Alps (17.491-503). Here, the previous triumphs of the Carthaginians are directly connected to their deaths. We have already seen how specific forms of nature, such as mountains and rivers, can mark the limit between empires, and between inappropriate and appropriate behaviour. Moreover, the triggering of non-natural events is also a kind of boundary crossing, as these phenomena profane sacred spaces and gods, and interrupt the usual order of nature. Both Hannibal and Scipio (and Regulus, in the past) are 45 guilty of causing non-natural events, but the cycle halts with Scipio Africanus, that paradigm of Roman virtue, who implicitly denounces such treatment of nature at 13.741-3 (quoted earlier). The character of anyone who crosses these boundaries is immediately called into question, but Hannibal, who transgresses nature’s boundaries at every stage of his campaign, is more excessive, impious and immoral than anyone else. His failures are also correspondingly greater.50 Rivers and other waterscapes are of particular importance to the Punica. While other aspects of nature, such as mountains, forests and fields, do feature in the epic, they appear less frequently than rivers. The historical record dictates that nearly all the major Italian battles of the Second Punic War occurred near or beside bodies of fresh water, but all the same, Silius makes much more out of this than a straight versification of the historical facts would require, by personifying the rivers, exaggerating their pollution, and referring back to the abuse they experienced. Beyond the major martial rivers of the Punica, other waterscapes appear frequently in catalogues, often denoting the homeland of an individual or group. Rivers are especially prominent in the catalogue of Roman forces in book 8 (356-616). As a result, destruction of riverine environments has potential implications not only for the ecological landscape, but for the local communities as well. But rivers can also be politically charged; we have already seen how the crossing of certain rivers could act as a political statement, or an expression of imperialism. g ( ) 51 Coinage was an especially popular medium for displaying riverine collaboration and subjugation. See Campbell (2012) 378-383. 50 Indeed, Hannibal’s inability to recognize boundaries present in nature prefigures his failure to overcome the walls of Rome, which are the ultimate boundary, and which Jupiter is unwilling to cede. See von Albrecht (1964) 24-46. at miles dubio tardat vestigia gressu, More than this, rivers were often depicted as being complicit and involved in these processes.51 In the Punica, the major rivers are all Italian, and most are shown to be the unwilling victims of a Carthaginian assault, mirroring Roman sentiment. The Trebia is exceptional; rather than remaining neutral as most of the other rivers do, it acts in accordance with Juno’s wishes, and therefore supports the Carthaginians. The river subverts the political status quo, which dictates that the rivers of Italy ought to be loyal to Rome alone. Indeed, Scipio’s major gripe with the Trebia is that it has become Carthaginian despite existing on Italian land (4.643-8). It is for this treachery that he threatens to obliterate the Trebia altogether. There exists a tension between the idea that domesticated rivers should help, or at least refuse to harm, those who acted as their 46 domesticators, and the fact that, in reality, rivers are unpredictable and often their natural processes are destructive to the communities and empires who lay claim to them. Even the Tiber flooded frequently, despite being Rome’s tutelary river.52 The Trebia episode partly serves to make sense of this contradiction: the river floods and attacks the Romans because it is obedient to Juno, and by extension the Carthaginians, but also because rivers do not always perform in the way one might expect; variability is fundamental to their phusis. Conversely, mountains are solid masses of unchanging rock. They too are important to the Punica as that other major physical manifestation of boundaries, yet their role is much reduced compared to that of the rivers. Unlike rivers, which are fluid and permeable, mountains are immovable and inflexible. Personification of mountains requires far more imagination than personification of rivers, which are already associated with life, movement and changeability,53 and indeed, the Alps are not personified in the Punica. Landslides and avalanches make mountains potentially dangerous, but they cannot move from their position, unlike rivers, which can flood and find new avenues in which to flow. Because of the immovability of mountains, and their immensity that makes them difficult to alter and control, they cannot exemplify the domesticated/undomesticated/natural/non-natural spectrum as clearly as rivers can. The principal attitude towards nature in the Punica is pragmatic; Silius portrays nature primarily as a marker of boundaries, of both the physical and metaphysical kinds. 52 See Aldrete (2007) on the flooding of the Tiber. Of particular interest is the table on page 15 15. 53 See Clarke (1997). at miles dubio tardat vestigia gressu, This is by no means a new use of nature in epic, as Lucan’s Civil War demonstrates, but it does suggest an increased understanding of nature as something partly inviolate, which should, perhaps, be left to its own devices. However, the focus on Italian nature, and its frequent victimisation at the hands of the Carthaginians, suggests that nature is also seen to be part of Rome’s political economy; the destruction to the Italian landscape is, on the face of it, an environmental catastrophe, but more importantly, it is an assault on the Roman Empire itself. Furthermore, the Romans maintain careful control over those environmental borders that separate them from their enemies, as the images paraded in the triumphal procession of the final book illustrate: 52 See Aldrete (2007) on the flooding of the Tiber. Of particular interest is the table on page 15 47 terrarum finis Gades ac laudibus olim terminus Herculeis Calpe Baetisque lavare solis equos dulci consuetus fluminis unda, frondosumque apicem subigens ad sidera mater bellorum fera Pyrene nec mitis Hiberus cum simul illidit ponto, quos attulit, amnes (17.637-42). Gades at the ends of the earth, and Calpe, the limit of ancient Hercules’ accomplishments were both there. And the Baetis too, who is accustomed to bathing the sun’s horses in the waves of its sweet stream. Also wild Pyrene, the mother of war, rearing her leafy peaks to the stars, and the Ebro, no mild river when it guides all those streams that is has brought with it into the sea. Two of those boundaries mentioned are cities: Gades in south-western Spain, and Calpe, the limit of Hercules’ achievements. The rest – the Baetis, the Pyrenees and the Ebro – are all environmental borders. With the end of the Second Punic War, some of the boundaries over which Hannibal crossed – namely the Pyrenees and the Ebro – are symbolically restored to Roman rule. However, complete authority over nature, even if it is Roman authority, is not necessarily a positive thing; the discomfort evinced by the elder Scipio’s threats to annihilate the Trebia indicates that there is tension between human technological mastery and the power it can wield over nature, especially given that nature is often partly divine. 54 See Campbell (2014) 372: “It was reasonable that river-gods too, once won over by imperial might and justice, would lend their efforts to supporting the emperor.” at miles dubio tardat vestigia gressu, For this reason emperors were anxious to be seen as cooperating with rivers, rather than exerting brute force over them.54 This attitude exists within the Punica, and some of the major characters subscribe to it while others do not: Scipio, Regulus and Hannibal all treat nature with some measure of disrespect, and this contributes to their status as liminal figures. On the other hand, Scipio Africanus recognises the importance of protecting nature from exterior forces, as we saw in the katabasis of book 13, and this adds gravity to his ultimate success – the capture and defeat of Hannibal. 48 Overall, nature within the Punica, in addition to its role in the historical record, has a primarily moralising function. While there is concern over pollution and the effects of non-natural phenomena, that anxiety is not explicitly environmental. Of greater concern is the impiety and moral liminality associated with boundary- crossing, and the threat that boundary-crossing, exemplified by, but not limited to, non-natural phenomena, poses to Roman imperial stability. The ecological violations that transpire in the epic are, arguably, unavoidable symptoms of war, but the significant imbalance between (minor) Roman and (major) Carthaginian responsibility aids in the vilification of Rome’s enemies, and suggests a moral superiority on the part of the Romans, who very rarely transgress boundaries in the epic, and who are characterised by virtus and fides. Silius’ Punica is by no means categorically pro-Roman, but it is, nevertheless, a celebration of Roman achievements, and equally a condemnation of Carthaginian exploits, as the very first invocation illustrates: da, Musa, decus memorare laborum antiquae Hesperiae, quantosque ad bella crearit et quot Roma viros, sacri cum perfida pacti gens Cadmea super regno certamina movit (1.3-6). da, Musa, decus memorare laborum antiquae Hesperiae, quantosque ad bella crearit et quot Roma viros, sacri cum perfida pacti gens Cadmea super regno certamina movit (1.3-6). Allow me, Muse, to recount the glory of Italy long ago, and to speak of how many great men Rome produced for war, when the Carthaginians betrayed the sacred agreement and set in motion a struggle for power. From the first, Rome is already set high above Carthage in terms of virtus, and the rest of the epic exposes this disparity. 1 The relative chronology of these two texts is uncertain. Ripoll (2015) consolidates all the evidence but comes to no firm conclusions. However, there is some suggestion that Silius wrote his mache parapotamios before Statius wrote his; see Wistrand (1956) 58-9 and Dewar (1991) xxxi. 2 at miles dubio tardat vestigia gressu, Nature, too, plays a crucial role: a man’s virtue in the Punica may be measured, at least in part, by the damage he inflicts upon nature, which generally does not deserve the violent treatment it so often receives. 49 ( ) 3 Like Silius, the Homeric source material informs much of Statius’ own mache parapotamios. See Juhnke (1970) 24-44. 2 See Conrau-Lewis (2014). 3 3. Thebaid 9: Hippomedon and the Ismenus 3. My third and final chapter concerns a second Flavian epic, Statius’ Thebaid. This poem narrates the struggle between Eteocles and Polynices for the throne of Thebes. It begins with the exile of Polynices after his brother, Eteocles, takes the throne. Filled with resentment for his brother’s actions, Polynices assembles an army and marches against Thebes. At the climax of the narrative, the two brothers fight one-on-one and kill each other. The epic concludes with funeral fires for the major warriors, and the appearance of Theseus, who attempts to restore order, inciting a second civil war in the process. The Thebaid differs in many ways from its contemporary, the Punica,1 but it too features a mache parapotamios: in book 9 the Argive hero Hippomedon, our Achilles/Turnus figure,2 has his aristeia, the climax of which is a battle with the Ismenus River.3 The battle in the river begins at 9.225 when the fighters reach the riverbanks. The weight of the men at the river’s edge causes the banks to give way (230-1) but Hippomedon jumps in anyway (233). Scenes of chaos follow, in which men struggle in the waves, many drowning as they try to hide or escape from Hippomedon (236- 41). A brief interlude interrupts the carnage to introduce a new warrior – Crenaeus – who is the grandson of the Ismenus (319-37). Hippomedon kills the young warrior (345-6) and he is mourned by the waves and surrounding woods horruit unda nefas, silvae flevistis utraeque, \| et graviora cavae sonuerunt murmura ripae (“the waves shuddered at this monstrous crime, the woods on either side wept, and the caved-in banks resounded with a louder rumbling,” 347-8). After Crenaeus’ death, his mother – the nymph Ismenis – seeks out his body and rebukes the Ismenus for failing to look out for the boy (351-98). Ismenus is completely unaware of what has been occurring in his own stream: at pater arcano residens Ismenos in antro (“but Father Ismenus was resting in his private hollow,” 404) and he only learns of Crenaeus’ death after a nymph tells him of it (416-8). When he discovers what has been occurring in his 50 stream, Ismenus complains that he is inundated with bodies, weapons, and gore (429- 33). He then rebukes Hippomedon (442-5) and begins an assault against the warrior (446-506). Ganiban (2007), especially 1 2. 6 See Feeney (1991) 354; Ganiban (2007) 50-5; McNelis (2007) 6-7 and Micozzi (2015) 326- 7. For a gender-specific interpretation of the Aeneid’s influence on the Thebaid, see McAuley (2016) 368-73. 4 Of further interest on Ovidian influence in the Thebaid is Keith (2004/5). 5 5 Ganiban (2007), especially 1-2. 6 7 See Schildgen (2013) 88-90. 3. Thebaid 9: Hippomedon and the Ismenus Eventually Hippomedon appeals to the gods for help (506-10) and Juno asks Jupiter to take action (510-19). With a nod of Jupiter’s head, the river subsides (520-1). In my previous two chapters I discussed the presence of nature in the Iliad and the Punica, and in both cases concluded that while the destruction of nature is a major concern in the texts, it is not necessarily an environmental matter, but rather a religious, cosmological, and political one. Nevertheless, it is possible to extract some underlying attitudes towards nature; for example, our texts suggest that nature is sacred, mysterious, and an integral part of the cosmic order. At the same time, it is valued according to how helpful or harmful it may be to humankind. In this chapter I will use the proposed schema to argue that the Thebaid treats nature in a similar way. I will also show, however, that within this framework the Thebaid differs in some crucial ways from the Iliad and the Punica. First, let us take a look at the influence of Statius’ predecessors on his own work. Much of what has been said regarding Silius and the influence of Virgil, Lucan and Ovid on the presentation of rivers in the Punica applies also to Statius’ Thebaid and does not require repeating (see Chapter 2 above).4 However, the overall influence of the Aeneid on the Thebaid should not be overlooked, as Statius’ narrator famously says at the end of his epic: vive, precor; nec tu divinam Aeneida tempta, \| sed longe sequere et vestigia semper adora (“Live, I pray, and do not attempt to be the divine Aeneid, but follow it at a distance and always revere its footsteps,” 12.816-7). As Ganiban argues, Statius does not try to outdo Virgil, but rather criticises and reinterprets the Aeneid from a moral and political perspective.5 Strikingly, Statius reinterprets the roles of the gods using a Virgilian paradigm; he assigns Jupiter the role of vindictive and wrathful god that had previously characterised the Juno of the Aeneid, and in turn, emphasises the change by likening Juno to the Virgilian Jupiter, insofar as she is a (weak) voice of reason.6 In terms of the landscape, Statius rejects 51 Virgil’s ambiguous treatment of the natural world. Here, a passage from the Aeneid describes the felling of a grove for the construction of Misenus’ pyre: Virgil’s ambiguous treatment of the natural world. 3. Thebaid 9: Hippomedon and the Ismenus Here, a passage from the Aeneid describes the felling of a grove for the construction of Misenus’ pyre: itur in antiquam silvam, stabula alta ferarum; procumbunt piceae, sonat icta securibus ilex fraxineaeque trabes cuneis et fissile robur scinditur, advolvunt ingentis montibus ornos (6.179-82). Into the ancient forest they go, the dwelling place of wild animals. Pines fall, while the pierced ilex resounds with each blow of the axe. Ash and oak trees are split in two with wedges, and they roll the mountain ash down the steep slopes. This passage is reconstructed in the Thebaid.7 In particular, note the corresponding use and positioning of procumbunt piceae, fraxineae/fraxinus and scinditur at the beginning of their lines, as well as robur at the end: …cadit ardua fagus The lofty beech tree falls, as well as the Chaonian wood and the Cypress that can withstand the winter. The pines fall, food for high flames, and so do the mountain ash and oak trees, and the yew with its treacherous sap, and ash that will drink the blood of unspeakable warfare, and unassailable oak. Then the spirited fir tree falls, and the 7 See Schildgen (2013) 88-90. 52 pine is split with a fragrant wound, and the alder, a friend of the sea, and the elm who is not inhospitable to vines, lean their clipped tops on the earth. Virgil’s description of the felling of the grove generates ambivalent anxiety around the encumbrance of civilisation on the landscape. 8 Statius takes a more explicitly moralising approach to the felling of the Nemean grove, denouncing it as a sacrilegious act (see pp. 9-11 above). The Thebaid challenges Virgil’s comparatively indifferent approach to the violation of the landscape at the hands of Aeneas and his men by clarifying and underlining the moral issues at play and explicitly problematizing humankind’s relationship with the landscape.9 The Aeneid, therefore, partially informs an understanding of nature in the Thebaid. It would be remiss, however, to omit mention of the influence of Callimachus, in particular where rivers are concerned. ( ) 9 This passage also has a precursor in Lucan, whose description of the felling of the Massilian grove (3.399-445) involves similar desecration, although even here the crime is met with an ambiguous, even cynical, observation about divine retribution: servat multos fortuna nocentes: \| Et tantum miseris irasci numina possunt (448-9). See Augoustakis (2006a). \| p ( ) g ( ) 10 I.e. that Callimachus was anti-epic, and critical of narrative digression, which could lengthen a poem unnecessarily. This is a contested interpretation of Callimachean poetics; Cameron (1995) 403-7 argues that Callimachus’ Hymn to Apollo is not anti-epic, but critical of elegy that attempts to imitate epic style. 11 gy p 11 McNelis (2007) 87. 8 Thomas (1988) 267-8. 8 Thomas (1988) 267-8. 9 ( 12 Ibid. 112-5. 8 Thomas (1988) 267-8. 9 This passage also has a precursor in Lucan, whose description of the felling of the Massilian grove (3.399-445) involves similar desecration, although even here the crime is met with an ambiguous, even cynical, observation about divine retribution: servat multos fortuna nocentes: \| Et tantum miseris irasci numina possunt (448-9). See Augoustakis (2006a). 10 I.e. that Callimachus was anti-epic, and critical of narrative digression, which could lengthen a poem unnecessarily. This is a contested interpretation of Callimachean poetics; Cameron (1995) 403-7 argues that Callimachus’ Hymn to Apollo is not anti-epic, but critical of elegy that attempts to imitate epic style. 11 McNelis (2007) 87. 12 Ibid. 112-5. 13 Ibid. 137. 13 Ibid. 137. 3. Thebaid 9: Hippomedon and the Ismenus McNelis’ study of Callimachean intertextuality in the Thebaid addresses Statius’ use of rivers, especially at Nemea, the Asopus, and the Ismenus, and suggests that they both reflect and resist Roman- Callimachean poetics.10 For example, the drought in book 4 signals a “counter to the poetic agenda” as it prevents the progression of the Argive troops, and therefore halts the continuation of the narrative.11 Furthermore, the Asopus, which takes its origins from Callimachus’ Hymn to Delos 75-8, is simultaneously anti-Callimachean, as the excursion into its past at Thebaid 7.317-27 delays the narrative further.12 The swollen Ismenus, which joins together with the equally elevated Asopus, is Callimachean in physical form, but abandons “Callimachean values of small-scale streams and poetics” as a result of its grandiosity.13 Chaudhuri also comments on the Callimachean elements of the mache parapotamios, suggesting that Statius employs Callimachean 13 Ibid. 137. 53 imagery in order to compete with, and even outdo, Homer. 14 Hippomedon’s exceptional display of strength, and the river’s corresponding intensity, challenges the Homeric notion that humans are powerless against the gods.15 Newlands has already made much headway on landscape in Ovid’s Metamorphoses and Statius’ Thebaid. Newlands examines three episodes in the Thebaid that demonstrate Statius’ treatment of the landscape. These are, in order of discussion: the sacred grove of Diana (4.419-42), the sacred grove of Nemea (described at various points in books 4, 5 and 6), and the River Ismenus (see above). 14 Whose poetry, some believe, is represented by the expansive πόντος in Callimachus’ Hymn to Apollo. See Williams (1978) 85-9, and Kahane (1994) 121. Others, such as Cameron (1995) 403-7, reject the association of πόντος with Homer. 15 Ch dh i (2014) 213 4 21 I.e. Hannibal, Scipio, Regulus. ( ) 16 Newlands (2004) 137. 17 ( 17 Ibid. 137-8. 18 ( ) , j 15 Chaudhuri (2014) 213-4. 18 Ibid. 136. 19 Ibid. 147. 15 Chaudhuri (2014) 213-4. 16 ( ) 24 Or, perhaps, flees from its own waters, see Smolenaars (1994) 382 on lines 800-1. This episode recalls the autochthonous origin of the Thebans, who celebrate this katabasis by recounting the story of Cadmus (8.218-39); see Keith (2000) 60-1. On Amphiaraus, see Vessey (1973) 258-69, Masterson (2005) and McNelis (2007) 127-30. 3. Thebaid 9: Hippomedon and the Ismenus All of these loci amoeni have some parallel in the Metamorphoses, but Newlands argues that while Ovid’s pleasant landscapes are deceptive and complicit in strife, the landscapes of Statius are victims of human aggression.16 Ovid’s sacred places are maintained by the divinities, but Statius’ gods are conspicuously aloof in this regard; they neither protect nor restore the earth.17 Newlands’ most significant point is that “landscape thus provides a significant moral and political crux in its interrogation of divine and human action.”18 Rivers demonstrate this especially well, as the watery landscape acts as a “symbolic topography in which the moral and physical evil of civil war is dramatically displayed in the sullying and swelling of formerly pure and peaceful waters.”19 Overall, Statius’ depiction of nature unambiguously condemns human violence and chaos, and the metamorphosis of the landscape from the locus amoenus into a polluted and violated space reflects the confusion and discord inherent to the civil war.20 Statius, then, takes a moralistic approach to violated landscape in the Thebaid. This is similar to what we have seen in the Punica, where the violation of nature is a transgressive and sacrilegious act. However, the Punica far more frequently pins the responsibility for the violation of nature on specific people,21 whereas the Thebaid 20 Ibid. 153-4. 54 utilises the motif to reveal the negative impacts of the civil war as a whole.22 I will return to this point in due course, but for now I will outline instances of domesticated, undomesticated, and non-natural nature in the Thebaid. Overall, nature in the Thebaid occupies a middle ground between nature in the Iliad and in the Punica. Nature is certainly much more present than in the Iliad and takes an active role in the narrative on a number of occasions. However, mentions of nature are less emphatic than in the Punica, where the pollution of the Italian rivers and the crossing of natural boundaries serve as important recurring thematic devices. Overall, nature in the Thebaid occupies a middle ground between nature in the Iliad and in the Punica. Nature is certainly much more present than in the Iliad and takes an active role in the narrative on a number of occasions. However, mentions of nature are less emphatic than in the Punica, where the pollution of the Italian rivers and the crossing of natural boundaries serve as important recurring thematic devices. y ( ) ( ) ( ) 25 6.845; 7.217; 8.330; 10.88; 11.466, 607; 12.561, 645. There are two references to Natura in the Punica (11.187 and 15.75), both concerning humankind’s mortality. Notably, there are no references to Natura in the Aeneid. 26 22 And because the Thebaid is a part of the totalizing, archetypal genre of epic, the Theban civil war is paradigmatic of civil war in general. Therefore impressions of the civil war of the poem may also be impressions of all civil wars. 23 26 See Feeney (1991) 364-91. 23 Newlands (2004) 134. p y p 23 Newlands (2004) 134. 3. Thebaid 9: Hippomedon and the Ismenus Two instances of nature mentioned above – the Nemean grove and the Ismenus – are the longest and most significant treatments of nature in the epic. Nature appears on other occasions also: early on in the epic, after Polynices’ banishment, a ferocious storm wreaks havoc across the landscape (1.346-63), foreshadowing the turmoil to come.23 Other mentions of nature are brief but still significant: twice Dirce’s spring is described as running with blood (1.38; 4.374-5); Acheloüs makes an appearance in the catalogue of book 4, hiding his one-horned head under the waves while anhelantes aegrescunt pulvere ripae (“his panting riverbanks grow sick with dust,” 4.106-9). In book 7 the earth cracks open and swallows Amphiaraus, who descends to the Underworld while the Ismenus runs through new openings in its banks (794- 823).24 One ought also to note the frequent references to the personified goddess Natura, who is invoked or referenced some eight times in the epic.25 Her sphere of influence relates primarily to phusis (Latin natura) as we have so far defined it, although she is also at times associated with the physical world in general (e.g. at 10.88). The relatively frequent references to Natura are not especially surprising, given Statius’ penchant for personification,26 but they do perhaps suggest a greater connection between phusis and the divine than in other texts. 55 In the narrative of the Thebaid there are no conventional instances of domesticated nature. Unlike both the Scamander and the Trebia, who join forces with the Trojans and Carthaginians respectively, the Ismenus is not allied with either the Argives or the Thebans in the war, and does not actively support either group: amnis utrimque timet, crasso vada mutat uterque \| sanguine (“the river fears both, and both dye the shallows with foul blood”, 9.257-8). However, the possibility for domestication is manifest in the youthful Crenaeus, the grandson of the Ismenus: gaudebat Fauno Nymphaque Ismenide natus maternis bellare tener Crenaeus in undis, Crenaeus, cui prima dies in gurgite fido et natale vadum et virides cunabula ripae, ergo ratus nihil Elysias ibi posse Sorores, laetus adulantem nunc hoc, nunc margine ab illo transit avum: levat unda gradus, seu defluus ille, sive oblicus eat; nec cum subit obvius, ullas stagna dedere moras pariterque revertitur amnis (319-27). 3. Thebaid 9: Hippomedon and the Ismenus Tender Crenaeus, born of Faunus and the nymph Ismenis, was delighted to fight in the maternal waves, in whose eddies he had trusted since his first day, nurtured in the shallows and the cradle of green banks. Thus he believed that the Elysian Sisters were powerless there, and joyfully he crossed over his fawning grandfather, now here, now to the very edge. The waves rose to meet his steps, whether he went downstream or to the side. Even when he suddenly met the current head-on, none of the pools hesitated, and the stream followed suit and reversed. Here we have the youthful descendant of a river who faces a warrior much greater than himself, Hippomedon. The source model for Homer’s Asteropaeus is Crenaeus, the grandson of the Axius River whom Achilles fights and kills in the Scamander at Iliad 21.140-82. It is apparent, however, that Crenaeus is exceptional. Not only is he descended from the river, but the river actually flows in tandem with his movements, allowing him to walk on the waves, and even running in reverse if he should wish to 56 move upstream. The river’s cooperation with Crenaeus is a kind of proto- domestication that hints at possible past or future domestication. The river is only sympathetic to one individual, however, and though Crenaeus fights on behalf of the Theban host, the river cares only for him, not his allies. Domestication, as we have thus far seen it, is usually the product of close proximity between a group of humans and a specific river, as with the Trojans and the Scamander, or else a product of imperialism: the rivers of Italy in the Punica, for example, belong to the Roman political economy as they are located within the bounds of the empire. In the Thebaid, we would expect the Ismenus to be sympathetic to the Thebans since it is also located in Boeotia, east of Thebes, and historically provided a significant portion of the city’s water.27 Indeed, the Thebans do try to use the river as if it is domesticated: at 4.414-8 Eteocles undergoes a purification ritual in the Ismenus. 29 Compare, e.g. Neptune directly controlling the sea and winds at Odyssey 5.291-8. Statius’ indirect domestication here closely resembles Juno appealing to Aeolus to let loose the winds at Aeneid 1.64-86. For an alternative interpretation of this passage that focuses on gender, see Keith (2000) 57-60. Thebes, e.g. Euripides, Bacchae 5. 28 Dee (2013) 185-6. 29 27 Symeonoglou (1985) 9; 302. Elsewhere both the Ismenus and Dirce are synecdoche fo Thebes, e.g. Euripides, Bacchae 5. ebes, e.g. u p des 28 Dee (2013) 185-6. 29 3. Thebaid 9: Hippomedon and the Ismenus The ritual does not succeed and Eteocles remains impure; in any case, the Ismenus is an unsuitable agent of purification, as it will itself be polluted beyond recognition in the near future.28 The events at Nemea are also worth noting here: as the Argive warriors march towards Thebes, Bacchus orders all the streams in their path to dry up (4.684-96). Rather than appealing to the river deities directly, as we might expect, Bacchus asks the nymphs to activate the drought. The nymphs can control the water of the streams and hasten to obey Bacchus’ commands (697-710). This looks like domestication, but it is complicated by the indirectness of communication: Bacchus cannot control the rivers himself, nor does he command the rivers or river-gods directly; he liaises with the nymphs to achieve his end.29 This is much more discursive than Silius’ blunt introduction to his mache parapotamios: Trebia… precibus Iunonis suscitat (“The Trebia… roused by Juno’s request,” 4.573). It is clear here that the rivers themselves are not domesticated by Bacchus, although they may be domesticated by the nymphs if we consider divine control over nature to be domestication. However, Statius’ description of the nymphs suggests that they are assimilated with the rivers in a similar way to river-gods: ast illis tenvior percurrere visus \| ora situs, viridisque comis exaruit umor. \| protinus Inachios haurit sitis ignea campos (“A fine layer of 27 Symeonoglou (1985) 9; 302. Elsewhere both the Ismenus and Dirce are synecdoche for Thebes, e.g. Euripides, Bacchae 5. 28 57 dust seemed to pass over their faces, and green moisture dried out of their hair. Immediately, a burning thirst drains the Inachian fields,” 4.697-9). The nymphs do not just control the rivers; they are intimately connected with them, and they share the same experiences, more so, even, than river gods, who, if the deity of the Ismenus is anything to go by, are startlingly disconnected from what happens in their own waters. , ,g 32 See Dee (2013) 187: “whereas a living Eteocles is washed in a pure river, a dead Polynices (with Eteocles’ pyre still burning nearby) is being cleansed in a polluted river.” p 31 See, e,g. the list of omens at 7.402-23. 30 As the Tiber of the Aeneid is separate from Tiberinus. 31 30 As the Tiber of the Aeneid is separate from Tiberinus. 31 p 31 See, e,g. the list of omens at 7.402-23. 3. Thebaid 9: Hippomedon and the Ismenus Unlike both the Iliad and the Punica, whose river deities are either the same as the physical stream itself, or else so closely connected to the water that the two cannot be separated, the rivers and river-gods of the Thebaid are separate entities that can act independently of one another.30 This brings even further ambiguity to the idea of domestication in the Thebaid, and raises an important question: is the imposition of the river-god’s will (and the will of the nymphs) on the river an act of domestication, or is it simply a personified manifestation of a twofold, but ultimately homogenous, consciousness, like a form of double motivation? This is a question that I will return to later, but for now, suffice to say that the domestication of nature is much more ambiguous in the Thebaid than in either the Iliad or the Punica. The lack of successful or straightforward domestication in the Thebaid is unsurprising. The narrative orbits around the confusion and horrors of the civil war, where the usual order of things is destabilised. Nature, too, is thrown into confusion,31 and just as man’s relationship with his fellow man disintegrates in civil war, so too does man’s relationship with nature. Indeed, after the death of the brothers, two women, Antigone and Argia – one Theban, one Argive – come together to wash the corpse of Polynices in the Ismenus. Where before the river had failed to cleanse the body of Eteocles, here the body is restored: ut sanies purgata vado membrisque reversus \| Martis honos (“when the gore had been cleansed in the pool, the glory of war returned to his limbs,” 12.416-7). Only after these two women have introduced the possibility of reconciliation does the river return to a domesticated state and fulfil its purifying function. However, the success of the purification is called into question because the river is turpatus adhuc (“still polluted,” 410).32 The lack of successful or straightforward domestication in the Thebaid is unsurprising. The narrative orbits around the confusion and horrors of the civil war, where the usual order of things is destabilised. Nature, too, is thrown into confusion,31 58 Let us now turn to the non-natural phenomena present in the Thebaid. g 34 See Newlands (2004) 141-6 on Nemea. Also Parkes (2012) 322: “The Argive army displays its readiness to break out in civil conflict.” 35 3. Thebaid 9: Hippomedon and the Ismenus Like previous epic poets, Statius employs the trope of the river dyed with blood and clogged with bodies: ille ego clamatus sacris ululatibus amnis, qui molles thyrsos Baccheaque cornua puro fonte lavare feror, stipatus caedibus artas in freta quaero vias; non Strymonos inpia tanto stagna cruore natant, non spumifer altius Hebrus Gradivo bellante rubet (9.434-9). I, that sacred river echoing with frenzied cries, who am said to wash the pliant thyrsus and horn of Bacchus with my pure spring, I am packed with carnage and seek narrow paths to the sea. Not even the wicked pools of Strymon swim with such blood, and the foaming Hebrus is not dyed red to such a depth when Mars wages war. As in the other examples, the inundation displeases the river and it complains bitterly about the pollution, emphasising its previously pure (puro) state. Once again, the river’s thwarted volition indicates that a non-natural process is occurring. Pollution is the main form of non-natural phenomenon in the Ismenus episode, although it is not exclusive to the Ismenus. As I mentioned earlier, Dirce’s springs are twice said to run with blood (1.38; 4.374-5). However, river pollutants are not limited to blood and gore; dust and mud are also styled as pollutants on a number of occasions, most emphatically at Nemea, where the parched Argive soldiers descend upon the river: fremunt undae, longusque a fontibus amnis diripitur, modo lene virens et gurgite puro perspicuous, nunc sordet aquis egestus ab imis alveus; inde tori riparum et proruta turbant gramina; iam crassus caenoque et pulvere torrens, quamquam expleta sitis, bibitur tamen. agmina bello decertare putes iustumque in gurgite Martem 59 33 Vessey (1973) 170: “Water is an ancient image of life, and the thirst suffered by the warriors is a foreshadowing of their future death.” 34 p y 35 Newlands (2004) 145. p y 35 Newlands (2004) 145. perfurere aut captam tolli victoribus urbem (4.823-30). perfurere aut captam tolli victoribus urbem (4.823-30). The waves roared, and the river was torn far from its spring. Once a clear, translucent green with pure eddies, now the emptying channel was polluted all the way down to the bottom. From there the water was churned up by the ridged grassy banks. Now the river was thick with mud and rushing with dust, and although they’d satisfied their thirst, they continued to drink, so that you’d believe they were warring armies, joining a regular battle raging beside the whirling pools, or that a captured city was being destroyed by its conquerors. This scene foreshadows the battle of the Ismenus to come later, where two armies do fight in gurgite, but here mud and dust replace blood as the main pollutants.33 Confusion and disorder reigns, as a space that should renew life turns into a landscape of death (some soldiers are carried away by the water or are trampled as they enter, 4.813-6), chaos, pollution and war.34 The scenes of ecological destruction emphasise the brutality of the Argive soldiers even before they have joined in a real battle. Indeed, after the death of Opheltes, the men will fell the trees of the Nemean grove in the same way that they despoiled the river: like victorious soldiers recklessly plundering a captured city (6.113-7).35 In a reversal of the common Iliadic simile that compares warfare to nature, these similes instead imagine man’s interaction with nature to be like another war, with nature the overwhelming loser. In addition to pollution, there are two instances of the backwards-flowing river motif. This motif is much less prominent in the Thebaid than in the Punica, but it is equally alarming. At 7.405, rivers flowing backwards are included in a list of omens: sideraque adversique suis decursibus amnes (“planets and rivers turned away from their courses”). Rivers are here directly associated with celestial bodies, emphasising their connection to the cosmic order; their reversal is a sign that the universe is in a state of disarray as the armies march towards the battlefield. perfurere aut captam tolli victoribus urbem (4.823-30). Indeed, the other omens are all of a similar type: blood and stones replace rain (408), ghosts appear (409), 33 Vessey (1973) 170: “Water is an ancient image of life, and the thirst suffered by the warriors is a foreshadowing of their future death.” 34 60 oracles fall silent (410), Eleusis cries out in the wrong months (411-2), and Acheloüs is said to have lost his other horn (416-7). The second reference to reversed rivers also occurs in book 7; here Tydeus warns Polynices against approaching Eteocles’ camp, fearing that the Theban king will never let Polynices return once he has entered his domain: Inachus ante retro nosterque Achelous abibit (“sooner will our Inachus and Acheloüs flow in reverse,” 553).36 Tydeus uses the image of the river running backwards to describe something that he believes is unlikely to occur, apparently unaware that such an event has already happened. There are no other instances of river reversal in the Thebaid; the Ismenus, though clogged with corpses, never flows backwards except to support Crenaeus. Therefore the primary non-natural phenomenon of interest is the pollution of river water with bodies, blood and equipment, as well as dust and mud. As Newlands has already argued, the destruction of the landscape displays the inherent nefas of the civil war. Those who damage nature are cast in a negative light. This is especially true of Hippomedon, whose actions ultimately violate a sacred space and its divinity. Indeed, Hippomedon is portrayed as explicitly anti-river:37 one of his major appearances occurs in book 7, when he becomes the first of his men to cross the swollen Asopus river (430-5), which not only foreshadows his later battle with the Ismenus, but also confirms his unchecked impiety.38 Hippomedon expresses anti-river sentiment in other ways, most blatantly by killing the grandson of the Ismenus, and the retributive consequences of his arrogance are made clear at the time of his death, when another river descendant, Hypseus, strips his corpse (9.540-3). Asopian Hypseus proves his mettle in battle when he matches Hippomedon in the slaughter (253-6),39 although the two never meet in combat. Hippomedon was able to overcome the Asopus once, but at the end of his life, the son of that same river strips him of his armour, thus ending Hippomedon’s narrative with a tenuous sense of justice, though Hyspeus is killed in the next instant by Capaneus (552-6). ( ) 39 See also lines 9.568-9: hic ferus Hippomedon, illic non segnior Hypseus \| fletur. p y 37 Like Achilles, he is disdainful of the divinity of the river and doubts its strength: quove has traxisti gurgite vires, \| imbelli famulate deo solumque cruorem \| femineis experte choris, cum Bacchica mugit \| buxus et insanae maculant trieterida matres? (9.477-80). 36 Cf. Ovid, Metamorphoses 13.324: ante retro Simois fluet. See Smolenaars (1994) 249 for other examples of this adynaton. g \| ( ) 38 Conrau-Lewis (2014) 65-6. Like Achilles, he is disdainful of the divinity of the river and doubts its strength: quove has traxisti gurgite vires, \| imbelli famulate deo solumque cruorem \| femineis experte choris, cum Bacchica mugit \| buxus et insanae maculant trieterida matres? (9.477-80). 38 Conrau Lewis (2014) 65 6 perfurere aut captam tolli victoribus urbem (4.823-30). More troubling than Hippomedon’s crimes against the river, however, is the killing of Crenaeus, whose youthful naivety contrasts with Hippomedon’s reckless 36 Cf. Ovid, Metamorphoses 13.324: ante retro Simois fluet. See Smolenaars (1994) 249 for other examples of this adynaton. 61 brutality – a furor that he inherited from the recently deceased Tydeus, who died while consuming the head of Melanippus (8.751-66).40 Conversely, Crenaeus is tener and wrongly believes he cannot be killed in the familial waters of the Ismenus. His shield depicts the abduction of Europa (332-6), whose own misplaced trust in Jupiter (near water, no less) mirrors Crenaeus’ misguided confidence in the river’s ability to protect him.41 The killing of Crenaeus holds meaning on multiple levels. Before his death, Crenaeus gives a short speech, warning Hippomedon not to defile the sacred waters (9.340-3), but Hippomedon ignores the threat and, with little difficulty, runs Crenaeus through with his sword (345-6). This action is described as nefas (347) and causes the woods and waters to tremble and resound (347-8). Crenaeus’ words reinforce the sanctity of the river, and his death confirms that Hippomedon has lost all sense of piety. By killing Crenaeus, he insults the divinity of the river and thereby sets himself up for future punishment.42 Indeed, when the river-god finally hears of his grandson’s death, he sets out to avenge Crenaeus, and although the river does not strike the final blow, Hippomedon is killed the moment he escapes from the water (526-36). Hippomedon’s brutality is further emphasised by the pathos of the scene to follow, in which Crenaeus’ mother searches for his body amid the carnage wreaked upon the river by Hippomedon. Yet the apex of Hippomedon’s transgression comes after the river has begun its assault: Hippomedon asks, unde haec, Ismene, repente \| ira tibi? (“from where has this unexpected anger come, Ismenus?” 476-7). Despite Crenaeus’ earlier warning, the Argive warrior still does not realise that he has done anything wrong.43 Hippomedon’s capabilities extend beyond those of his Homeric counterpart, and even beyond those of Scipio;44 Hippomedon fares better than both previous warriors in terms of the mache parapotamios, but the severity of his crimes is likewise inflated; his nefas is undoubtedly the greatest of all three. His death also comes much sooner, and unlike Scipio, he fails to recognise the gravity of his transgressions. 40 Vessey (1973) 295. 41 Faber (2006) 110. 42 Dewar (1991) 118-9. 43 Ibid. 119 44 Chaudhuri (2014) 210. Newlands (2004) 152. 46 Dominik (1994) 103: “In no sense can their tragic deaths be said to have achieved anything remotely positive. This complete waste of human life bears testimony to the total futility of war.” 47 y ( ) 48 Newlands (2004) 145 argues that this startling inclusion of Roman-specific gods “brings Rome into the orbit of the Theban conflict.” Note the invocations to Pales in Georgics 1.3.1 and 294. 49 h (1988) 47 Vessey (1973) 170. 48 45 Newlands (2004) 152. 45 Newlands (2004) 152. 46 Dominik (1994) 103: “In no sense can their tragic deaths be said to have achieved anything remotely positive. This complete waste of human life bears testimony to the total futility of war.” 47 Vessey (1973) 170. 48 Newlands (2004) 145 argues that this startling inclusion of Roman-specific gods “brings Rome into the orbit of the Theban conflict.” Note the invocations to Pales in Georgics 1.3.1 and 294. 49 Thomas (1988). 47 Vessey (1973) 170. 52 The episode also draws inspiration from Ovid’s description of Cadmus’ fight with the serpent of Mars in Metamorphoses 3.31-100. Cadmus, like Hippomedon, hurls a large rock at the serpent but fails to hurt it without the use of a javelin. Unlike the Nemean serpent, however, the serpent of Ares is deliberately cruel. See also Brown (2016) 216. perfurere aut captam tolli victoribus urbem (4.823-30). The scenes in the Ismenus, therefore, reinforce the argument that crimes against nature are immoral because they reflect the eruption of discord in previously 62 peaceful spaces.45 As at the Nemean grove, where a tranquil, life-giving river is distorted by lust for war and pollution, so too is the Ismenus transformed from a nurturing stream into a place of death, pollution and vengeance. The emphasis on Crenaeus’ upbringing within the river makes the transformation even more pronounced. Furthermore, when Crenaeus dies he joins the ranks of other young boys who have died or will die in the war, including Atys (8.554-86) and Parthenopaeus (9.683-874), and whose deaths represent the indiscriminate slaughter and futility of war.46 Like these boys, whose inexperience and youthfulness make them poorly suited to the battlefield, nature in its ideal form – the locus amoenus – is also unsuited to war, and is ‘killed’ with the arrival of human conflict. We have already discussed the drought that leads the Argive soldiers to pollute the waters of Nemea, a non-natural event that is likewise transgressive, and foreshadows the deaths of those same soldiers.47 However, the destruction of nature at Nemea is not limited to this pollution; indeed, the soldiers later fell the trees of the grove to atone for the killing of a sacred serpent (6.84-7), trees that had previously been inviolate (incaedua, 90). The description of the grove that follows emphasises its primordial lifespan (93-6) and the sorrow of its destruction (aderat miserabile luco \| excidium, 96-7). As the trees fall, dat gemitum tellus (“the earth lets out a groan,” 107) and Roman gods – Pales and Silvanus – flee from the woods (110-3).48 The emphasis on the grove’s numinosity and heritage problematizes the Argives’ decision to fell it, and concludes an extended narrative that sees the entire Nemean grove, including the river and the trees, completely destroyed. Thomas argues that in the works of Virgil, the felling of trees is a potentially transgressive activity as trees can be numinous. Deforestation is often necessary for civilization, and aboriculture can make a tree more useful to humans; yet the memory of what has been lost – namely, divinity – as a result of humankind’s interference remains an uncomfortable reality. g , p y , 51 In some versions the serpent kills Opheltes deliberately: fragments from Euripides Hypsipyle suggest the serpent throttled the child (904-5). 50 Keith (2000) 58-59: “With Hypsipyle’s help, the Argive forces slake their thirst at the waters of Langia, while Opheltes by contrast, misses his nurse and her milk.” perfurere aut captam tolli victoribus urbem (4.823-30). 49 This idea is certainly present here, as Statius demonstrates how the need to fulfill 63 certain requirements of civilization through funerary rites can come into conflict with other spiritual and religious concerns. Indeed, the destruction of the Nemean grove is the final result in a causal chain that began with the drought induced by Bacchus. Following the drought, the Nemean soldiers encounter Hypsipyle, who leaves her infant charge alone in the grove while she leads the men to water.50 As a result, the men crowd the river and cause it to become polluted. Simultaneously, the drought provokes the thirst of a monstrous serpent, which roams the land looking for water (5.518-27); on its way it flicks its tail unwittingly (ignaro serpente, 539)51 at Opheltes, who dies instantly (538-40). When the soldiers find the dead child, they avenge his death by killing the serpent (558-78). However, the serpent was sacred to Jupiter, and it is mourned by nymphs, fauns, and the surrounding landscape. The serpent at Nemea closely resembles the serpent at Bagrada, killed by Regulus in the Punica.52 Indeed, the warrior who struck the fatal blow, Capaneus, nearly meets his end then and there: ipse etiam e summa iam tela poposcerat aethra Iuppiter et dudum nimbique hiemesque coibant, ni minor ira deo gravioraque tela mereri servatus Capaneus; moti tamen aura cucurrit fulminis et summas libavit vertice cristas (583-7). Jupiter himself had already requested weapons from high heaven, and not long ago rain clouds and storms were assembling. But the anger of the god was not so great and Capaneus was saved, deserving of a heavier blow. However, the blast of lightning that had been stirred up ran its course and struck the very top of his crest. ( ) 55 Ibid. 261: “Statius indicates the inherent criminality of the Argive cause.” 56 ( ) 58 Both child and serpent are given the epithet sacer: at 4.729 and 5.505 respectively suggesting, perhaps, that they are necessary sacrifices. 53 He is struck by lightning after attempting to challenge the gods (10.827-939). See McNelis (2007) 140-5. 54 56 The serpent can stretch itself all the way across a river and inhibit its flow (5.516-7). 57 Brown (2016) 221 56 The serpent can stretch itself all the way across a river and inhibit its flow (5.5 57 ( ) 54 See Ganiban (2013) 260-1. p 57 Brown (2016) 221. 58 perfurere aut captam tolli victoribus urbem (4.823-30). 64 Instead, like Regulus, Capaneus survives only to be singled out for a later punishment.53 After the death of the serpent, the Argives are forced to prepare a funeral pyre for the creature, yet as we have seen, even their attempt at atonement leads only to further destruction of the landscape.54 This chain of environmentally devastating events problematizes the war and, in particular, the Argives’ place within it; despite attempts to rectify mistakes they have made, the Argives are unable to atone successfully for their sacrilegious behaviour.55 Indeed, Brown compares the Argive army to the serpent: both are too immense to be accommodated by the grove and river56 and both cause devastation as a result of their raging thirsts.57 The Argive soldiers kill the serpent, forgetting the destruction that they have already caused, and oblivious to the fact that by killing the sacred serpent, they are no better than the serpent that has killed an innocent child.58 We have moved a little way from rivers, but these parallel episodes are equally as significant, and they reinforce a vital point: that violence towards nature in the Thebaid begets only more of the same violence, just as civil strife brings only further civil strife. In the Thebaid, humankind’s struggle to live alongside nature directly parallels humankind’s struggle to live alongside itself. It is not only humankind’s relationship with nature that is compromised in the Thebaid; nature’s usual coherence is affected by the cosmic imbalance that mirrors civil strife, and doubts are raised over its ability to control itself. The problem of domestication, explored in some capacity above, further emphasises the lack of stability; in particular, the undeveloped domestication of the Ismenus, and the disassociation between river and river-god, exhibit the disruption of order in the world of nature. Statius’ narrator tells us that the god of the Ismenus is residing in antro during the battle in the stream. Any knowledge of what has occurred does not reach him until after his grandson has already perished, when a nymph tells him what happened (9.416-7). Up until this point the river has been acting of its own accord, apparently without the knowledge of its deity; the river attempted to protect Crenaeus, 53 He is struck by lightning after attempting to challenge the gods (10.827-939). See McNelis (2007) 140-5. 59 See this passage especially: Ego sum pleno quem flumine cernis \| stringentem ripas et pinguia culta secantem, \| caeruleus Thybris, caelo gratissimus amnis. \| Hic mihi magna domus, celsis caput urbibus, exit (Aeneid 8.62-5). 60 ( ) p 60 Dewar (1991) 134-5. y ( ) 62 Feeney (1991) 345-6. ( ) 61 Feeney (1991) 345. 62 Dewar (1991) 134 5. 61 Feeney (1991) 345. 62 p ( ) 60 Dewar (1991) 134-5. 61 ( ) 61 Feeney (1991) 345. 62 perfurere aut captam tolli victoribus urbem (4.823-30). 54 See Ganiban (2013) 260 1 65 while the god remained entirely unaware that the youth was in any danger at all. This disassociation is problematic; in the Iliad and the Punica the river and river-gods are one and the same, or else so connected they cannot be distinguished. Statius, the exception, is perhaps here modelling his verses on Virgil, yet the prophetic Tiberinus could hardly be said to lack knowledge of what occurs in or near his own stream.59 If they must be distinct, then the relationship between river and river-god should be the ultimate expression of domestication, as in the Aeneid, yet the river-god’s ignorance provides an opportunity for the river to act without the deity’s direct influence. There is nothing sinister about the river’s actions, and they correspond to the river-god’s motivations (i.e. to protect Crenaeus), but the separation is nevertheless uncomfortable – it is hard to imagine that the river-god could remain oblivious to the battle raging in his stream, especially when it has such physical effects on the riverbanks and water, yet this is indeed the case. Dewar sees the separation of god and river as a primarily poetic choice, especially insofar as it distinguishes Statius from Homer and Silius.60 However, there is also intra-textual significance, as the troubling split is a continuation of the ambiguous domestication present at the end of book 4 involving Bacchus and the nymphs, and therefore it is a further demonstration of the cosmological aspects of the civil war. Moreover, Ismenus’ twofold personality is a furtherance of the structural failure that occurs within the “vertical scheme” of the universe as outlined by Feeney.61 The universe is divided into three levels: the Olympians, the human world, and the Underworld, and the focus of the narrative shifts between each level continually. Yet the anticipated Virgilian interaction between the three planes is disrupted, with humans interacting far more successfully with the Underworld than with the Olympians, whose king fails to act, or acts in ways unexpected.62 Similarly, the Olympian gods do not cultivate a relationship with the natural world, and thus they do not heal it when it is devastated. perfurere aut captam tolli victoribus urbem (4.823-30). Minor gods like rivers do not fit neatly into the three-tiered scheme, and accordingly they are split between human and Olympian realms: they are as detached from their physical streams as Jupiter is disinterested in 66 maintaining the human world, but they nevertheless care for certain humans, though they are powerless to help them in any meaningful way. However, there remains a clear separation between gods and humans, as revealed during the battle between Hippomedon and the Ismenus, in which the river emerges the clear victor despite Hippomedon’s valiant effort to resist the barrage of water. While Statius’ mache parapotamios generates an uneasiness surrounding the civil war for its catastrophic effect on the cosmos, the entire episode is also charged with pathos, with the death of Crenaeus emphasising the war’s brutality. The Ismenus’ obliviousness during the battle in which his grandson is killed heightens the pathos further, as his ignorance prevents him from stepping in to thwart Hippomedon. While Statius’ mache parapotamios generates an uneasiness surrounding the civil war for its catastrophic effect on the cosmos, the entire episode is also charged with pathos, with the death of Crenaeus emphasising the war’s brutality. The Ismenus’ obliviousness during the battle in which his grandson is killed heightens the pathos further, as his ignorance prevents him from stepping in to thwart Hippomedon. What is more, the answering grief of Crenaeus’ mother, Ismenis, punctuates the violent scene with lamentation, louder even than the rushing waters (9.407-8). Her wailing finally gains the attention of the river-god, who raises his head and drops the pine tree and urn he is holding (408-10). Maternal grief features prominently in the Thebaid and reminds the reader of the cost of war, particularly when the victim is young.63 Ismenis’ maternal grief intrudes upon Ismenus’ leisure, even when the din of the battle could not. 63 E.g.: 6.135-92; 9.608-39; 12.805-9. See Vessey (1973) 296. On motherhood in Statius, see McAuley (2016) 297-389. 64 Newlands (2004) 151. 63 E.g.: 6.135-92; 9.608-39; 12.805-9. See Vessey (1973) 296. On motherhood in Statius, see McAuley (2016) 297-389 g ; McAuley (2016) 297-389. y ( ) 64 Newlands (2004) 151. 63 E.g.: 6.135-92; 9.608-39; 12.805-9. See Vessey (1973) 296. On motherhood in Statius, se McAuley (2016) 297-389. 64 E.g.: 6.135 92; 9.608 39; 12.805 9. See Vessey (1973) 296. On motherhood i McAuley (2016) 297-389. y ( ) 64 Newlands (2004) 151. perfurere aut captam tolli victoribus urbem (4.823-30). Moreover, as Newlands points out, the grief of Ismenis in this moment overshadows and disrupts Hippomedon’s aristeia, as the narrative pans away from his rampage and follows Ismenis downstream as she seeks her dead child.64 When battle is resumed, now between river and warrior, the terms of combat have changed: Hippomedon must face a god who is motivated by revenge for the death of his grandson: at tu, qui tumidus spoliis et sanguine gaudes insontis pueri, non hoc ex amne potentem Inachon aut saevas victor revehere Mycenas ni mortalis ego et tibi ductus ad aethere sanguis (9.442-5). And you, proudly rejoicing in the spoils and blood of an innocent child, you will not return triumphant from this river to powerful 67 Inachus or savage Mycenae, not unless I was born of mortal blood and yours came from heaven. In both Homer and Silius, the partisan river rebukes the warrior primarily for polluting his waters, but while the Ismenus does complain about the state of the river, he has no allegiance to either side, and his main motive for attacking Hippomedon is his familial grief.65 Statius adjusts the Homeric source model so as to capitalise on the pathos of the scene: he makes Crenaeus the grandson of the present river, while Asteropaeus is the grandson of a foreign one. Thus the death of the youth generates an immediate emotional response from the Ismenus, whereas the Scamander is only indignant in a general sense. Indeed, Statius’ poignant approach to the mache parapotamios is unique; while an audience may feel indignation and shock at the treatment of the rivers, or descendants of rivers, in the Iliad and the Punica, the new and overtly emotive intrusion of familial grief in Statius’ narrative encourages us to reconsider Homeric ideas about warfare in which conflict is normalised; although the Iliad informs the presentation of Statius’ battle scenes, the philosophy behind the conflict is more Virgilian in its complexity.66 In Statius’ narrative, the realities of war induce pathos, but rather than render this pathos a mere unavoidable truth of war, as it is in Homer, the highly emotive take on warfare opens up a possible discussion as to whether the present conflict – that is, the civil war – should be tolerable. Non-natural phenomena, so prominent in Silius, are thus overshadowed by the grief of mother and grandfather. ( ) 68 And perhaps of Silius: see Dewar (1991) 102. ( ) 66 On pathos in Virgil, see Conte (2007). 67 67 See Dee (2013). 68 65 Dewar (1991) 134-5. 65 Dewar (1991) 134-5. 66 p g 67 See Dee (2013). 68 ( ) 66 On pathos in Virgil, see Conte (2007). 67 p g ( ) 67 See Dee (2013). 68 perfurere aut captam tolli victoribus urbem (4.823-30). However, pollution remains an important thematic element in the Thebaid as a whole,67 and here the narrative does not recoil from an opportunity to emphasise further the disastrous effects of the civil war. As we have already seen, like other rivers faced with non-natural pollution of blood, gore, and equipment, the Ismenus voices its displeasure at such impurities. Yet unlike the battles involving the Scamander and the Trebia, the mache parapotamios in the Thebaid does not end with the burning of the river by Hephaestus/Vulcan. Instead, Juno makes a plea to Jupiter, and the river abates with nothing more than a nod of Jupiter’s head (9.520-1). This is an obvious rejection of the Homeric source model,68 but the lack of 68 purifying fire is significant on another level also; in chapter 1, I discussed how the fires of Hephaestus purified the Scamander, ridding it of the major pollutants and restoring it to its original condition. In the Thebaid, no such purification occurs, meaning that the river remains in a non-natural state even after the waters have subsided. Indeed, even in book 12 the water is still polluted with the carnage of the battle (409-10). The lack of cleansing fire signals a perpetuation of the disorder that has characterised the narrative so far, and allows the non-natural phenomenon to leave an indelible mark upon the landscape that persists even after the war has ended. As such, the mache parapomatios and other nature-related episodes are one of the many ways in which the narrative fosters a negative impression of the civil war. Unlike the Punica, in which the violation of nature represents individual moral deficiencies, violence against nature in the Thebaid presents a much more generalised view of the conflict. In particular, the Ismenus’ ambivalence towards both sides in the fighting, and its primarily parental motives for joining in the battle, challenge the earlier depiction of rivers as seen in the works of Statius’ predecessors. Previously, rivers held some responsibility for the actions of the side they chose to support, and were therefore willing participants in warfare, whereas in the Thebaid they are no more than victims of human aggression and violence. At Nemea, the river is polluted and violated despite providing much-needed water for the thirsty soldiers. 69 Newlands’ argument that Statius removes Ovidian collusion and deception from the landscape and replaces it with innocent victimhood is especially relevant here. However, the collusion of the landscape is not only Ovidian, but Homeric, Virgilian, and Silian also. 70 Many deny that there is any such commentary present in the Thebaid. Among these is Vessey (1973) 63, who claims, “it would be wrong to see in the Thebaid any subtle political allegory.” For a comprehensive review of those who agree with this interpretation, see Dominik (1994) 131-2. For arguments that support the presence of political ideology in the Thebaid, see Dominik (1994) 132-80, McGuire (1997), and Braund (2006). ( ) ( ) ( ) 71 McGuire (1997) 94: “[The Flavian poets] frequently set their scenes of civil war in close conjuction to tyrannical figures and to suicidal actions, and they focus repeatedly on civil war’s capacities to obliterate more normal social, individual, and even linguistic traits.” 72 p , , g 72 Compare the references to gloria Aeneadum (1-2) and decus laborum Hesperiae (3-4) in Silius’ proem. By contrast, Statius says he will deliberately not address tales of Roman triumph (1.17-8). perfurere aut captam tolli victoribus urbem (4.823-30). Later, the Ismenus is attacked, polluted and desecrated despite having no specific ties to either the Thebans or the Argives, and, considering the inattentiveness of the river-god, no interest in fighting in a battle between them.69 Even when the god of the Ismenus does join the battle, it is not with the intention of swaying the war one way or the other, but simply to punish one man who killed an innocent member of the river’s family, while simultaneously committing a significant act of sacrilege. Thus I reiterate my original statement that Statius, like both Homer and Silius, does not approach the destruction and violation of nature as if it were an environmental problem. Rather, the ecological crises are indicative of a greater issue, namely the cosmological strife brought about by the civil war. Whether or not Statius 69 intended his epic to comment on the politics of his time,70 the poem as a whole presents civil war as a greater social, political and moral crisis than the foreign wars described in Homer and Silius.71 Indeed, there is no prediction of glory for Thebans or Argives in the proem of the Thebaid, only profanis odiis (“unholy hatred,” 1.1-2), and sontes Thebas (“guilty Thebes,” 2).72 Appropriately, scenes depicting the violation of nature in Statius are more shocking, more emotionally affecting, and more physically destructive than the equivalent scenes in the works of his predecessors. Furthermore, Statius challenges the ways we have been interpreting nature by introducing more ambiguity, particularly in terms of domestication, which complements the disturbed relationships between the Underworld, humans, and the Olympians. Just as the structure of the cosmos breaks down in this story of civil strife, so too does the natural/non-natural schema begin to unravel as the usual order of things goes awry. Statius simultaneously reinforces and reinvents the way nature is used in epic, distorting the usual associations between humans, gods, and the landscape found in Homer, Ovid, Virgil, and Silius; the newly fragmented terrain conveys certain ideas about civil war, namely that it adversely affects human relationships with other humans, gods, and the landscape, and causes terrible and unnecessary strife. The suffering of nature in the Thebaid, therefore, exemplifies the effects of the conflict, and invites the reader to acknowledge that there can be no victors in civil war. 70 Many deny that there is any such commentary present in the Thebaid. Among these is Vessey (1973) 63, who claims, “it would be wrong to see in the Thebaid any subtle political allegory.” For a comprehensive review of those who agree with this interpretation, see Dominik (1994) 131-2. For arguments that support the presence of political ideology in the Thebaid, see Dominik (1994) 132-80, McGuire (1997), and Braund (2006). 71 McGuire (1997) 94: “[The Flavian poets] frequently set their scenes of civil war in close conjuction to tyrannical figures and to suicidal actions, and they focus repeatedly on civil war’s capacities to obliterate more normal social, individual, and even linguistic traits.” 72 Compare the references to gloria Aeneadum (1-2) and decus laborum Hesperiae (3-4) in Silius’ proem. By contrast, Statius says he will deliberately not address tales of Roman triumph (1.17-8). perfurere aut captam tolli victoribus urbem (4.823-30). , ( ) , ( ), ( ) 71 McGuire (1997) 94: “[The Flavian poets] frequently set their scenes of civil war in close conjuction to tyrannical figures and to suicidal actions, and they focus repeatedly on civil war’s capacities to obliterate more normal social, individual, and even linguistic traits.” p , , g 72 Compare the references to gloria Aeneadum (1-2) and decus laborum Hesperiae (3-4) in Silius’ proem. By contrast, Statius says he will deliberately not address tales of Roman triumph (1.17-8). 70 Conclusion In book 23 of Nonnus’ Dionysiaca, a dying soldier addresses the swollen Hydaspes: προχοῇσι πόθεν σέο τέκνα καλύπτεις; πολλάκι Βάκτρον Ἄρηα µετήιον, ἀλλὰ ῥεέθροις οὔ ποτε Μῆδον ὅµιλον ἀπέκτανε Μῆδος Ἀράξης, Περσικὸς Εὐφρήτης οὐκ ἔκρυφε γείτονα Πέρσην: πολλάκι µοι παρὰ Ταῦρον ἔην µόθος, ἀλλ᾿ ἐνὶ χάρµῃ οὐ Κίλικάς ποτε Κύδνος ἑῷ τυµβεύσατο κόλπῳ οὐ Τάναϊς χιονῶδες ἄγων πετρούµενον ὕδωρ γείτονι Σαυροµάτῃ θωρήσσεται, ἀλλὰ κορύσσων Κόλχοις ἀντιβίοισι χαραδρήεσσαν Ἐνυὼ πολλάκι παχνήεντι κατεπρήνιξε βελέµνῳ. Ἠριδανὸς πέλε σεῖο µακάρτερος, ὅττι ῥεέθροις ἀλλοδαπὸν Φαέθοντα καὶ οὐκ ἔκρυψε πολίτην, οὐ Γαλάτην ἐκάλυψε καὶ οὐ τάφος ἔπλετο Κελτῷ, ἀλλὰ φίλοις ναέτῃσι ῥυηφενέων ἀπὸ δένδρων Ἡλιάδων ἤλεκτρα φεραυγέα δῶρα κυλίνδει: Ῥῆνος Ἴβηρ βρεφέεσσι κορύσσεται, ἀλλὰ δικάζων, καὶ κρυφίην ὠδῖνα διασχίζων τοκετοῖο κτείνει ξεῖνα γένεθλα (80-96). Why do you drown your own children in the water? I have fought in wars against the Bactrians many times, but the Median Araxes never killed a Median army in its stream, and the Persian Euphrates never drowned its Persian neighbours. I have often been to war beneath the Taurus, but even in the heat of war the Cydnus never made a tomb for the Cilicians in its lap, nor does the snowy Tanais ever arm itself with frozen water against the Sauromatans, but often attacks the hostile Colchians with torrential war and hurls frozen hail stones upon them. The Eridanus was more blessed than you when he drowned the foreigner Phaëthon in his stream, rather than one of his own people; he never drowned a Galatian, he never buried 71 a Celt, but he carries amber gifts of prosperity from the trees of the Heliades to his nearby friends. The Iberian Rhine attacks his children, but only to pass judgement; he marks out the child born in secret and kills the foreign offpring. Following a bloody battle in the Hydaspes, Dionysus’ forces begin their crossing of the river. However, at the behest of Hera, Hydaspes launches an assault on the army mid-way across, citing offence at being treated like a mere road (183). He sends a barrage of waves against Dionysus and his followers (192-214), and Dionysus responds with his own fury, censuring the river for displaying insolence towards a greater god (225-51). Then Dionysus lights the Hydaspes on fire (255-9), which burns even more violently than the Scamander and the Trebia did (259-79). Conclusion Hydaspes raises a plea to Dionysus, begging for mercy: ὕδασι καρποτόκοισι φέρων χάριν (“show kindness to my life-bearing waters!” 11). He confesses to being misled by a love for his children and submits himself to Dionysus, who removes the torch from the water (10-62). Before concluding this thesis, I will make a few brief remarks on the mache parapotamios of the Dionysiaca to illustrate some of the ways that the river battle has been standardised as a type scene in epic tradition before the Dionysiaca. Owing to space constraints, the Dionysiaca will not receive its own chapter. However, an overview of the passage will demonstrate the importance of the mache parapotamios for discerning perceptions about nature, humans, and the divine in epic, even a late- antique one such as the Dionysiaca. The soldier’s suggestion in lines 80-96 that no river should harm its friends or neighbours is a familiar refrain found in all three epics that I have discussed. At times, the rivers are resistant to this idea: the Trebia switches loyalties, fighting against the Romans with whom it ought to be allied, and the Ismenus chooses no side, fighting only for its family and not with its patron city, Thebes. Yet in these instances the rivers are not entirely guilty; their motivations are clear and quite understandable. The Trebia, though an Italian river, is defiled by the very people who are supposed to be its allies. Meanwhile, the Ismenus is embroiled in a civil war in which the leaders on both sides are Theban kings, and thus the distinction between Thebes and its enemies is not so clear-cut. The narrators of both epics justify the rivers’ behaviour and simultaneously suggest that humans have some responsibility in maintaining healthy 72 relationships with their rivers. Indeed, in the Iliad, the Trojans treat their river with respect, and it, in turn, offers them some protection. Meanwhile, Achilles, Scipio, and Hippomedon, who defile their respective riverine opponents, all emerge as morally ambiguous characters whose actions are problematic. However, in the Dionysiaca, the morality of Dionysus is never in question. Dionysus is a god whose authority is far greater than the river’s. Though he plays the part of Achilles, he also plays the part of Hephaestus, whose fires quell the rampaging river. 1 E.g. at 22.364-5, 370-1, 382-3, 23.4-6. Conclusion The river’s allegiance is to the Indians, its domesticators, but the Dionysiaca stresses that the river was wrong to favour its neighbours over the Olympian gods. Indeed, the river admits its transgression: 1 E.g. at 22.364-5, 370-1, 382-3, 23.4-6. ἀασάµην, Διόνυσε πυριτρεφές: οὐρανίην γὰρ σῶν δαΐδων ἀµάρυγµα τεὴν κήρυξε γενέθλην. ἀλλὰ πόθος τεκέων µε βιήσατο (24.13-5). ἀασάµην, Διόνυσε πυριτρεφές: οὐρανίην γὰρ σῶν δαΐδων ἀµάρυγµα τεὴν κήρυξε γενέθλην. ἀλλὰ πόθος τεκέων µε βιήσατο (24.13-5). I have acted foolishly, Dionysus, born of fire. For the glimmering of your torch heralds you as one born from heaven. But love for my children drove me to act as I did. The message is clear: though rivers are typically bound by their human domesticators, the gods ought always to be obeyed. There is little sympathy for the river, and indeed the occurrences of non-natural phenomena are downplayed compared to the three previous epics. Unlike the other rivers, the Hydaspes does not explicitly complain about the pollution of blood and bodies in the river, although pollution does indeed feature during the battle prior to the mache parapotamios.1 At the conclusion of book 22, a naiad emerges from the river and declares that she will leave for the untainted sea (392-401), but her chastising words against the pollution are not echoed by Hydaspes, who complains only that Dionysus treats him as if he were a road. Indeed, in a moment of dissonance the river exclaims: ὑγροβαφὴς δὲ \| Νηιὰς ἐν προχοῇσι πόθεν χρεµετισµὸν ἀκούει; (“why does the dripping naiad in my river mouth hear the neighing of horses?” 23.183-4) – a rather dissociative observation, given that the 73 73 naiad we heard from earlier fled because of the pollution, not the crossing of Dionysus’ army.2 In any case, despite the changes in structure and presentation of Dionysiaca’s river battle, the mache parapotamios remains the same at its core. All four examples, then, are more than simple displays of heroic prowess; they are also explorations of the relationships between humankind, nature, and the divine. In chapter 1, I discussed the mache parapotamios of the Iliad. I explained the differences between the presentation of nature in the Iliad and the Odyssey, noting that the Iliad presents a more positive view of domesticated nature as something that is unambiguously helpful to humans, while the Odyssey depicts domesticated nature as desirable, but not always virtuous. I also pointed out that undomesticated nature in the Iliad is dangerous, and also akin to war itself, while in the Odyssey it is a direct threat to the established peace. 2 See Schmiel (2003). ἀασάµην, Διόνυσε πυριτρεφές: οὐρανίην γὰρ σῶν δαΐδων ἀµάρυγµα τεὴν κήρυξε γενέθλην. ἀλλὰ πόθος τεκέων µε βιήσατο (24.13-5). Furthermore, I concluded that there is no moral code that brands Achilles as immoral because of his assault upon the river, despite its divinity, but that his actions are liminal because they disregard some social norms widely practised in the Iliad, such as piety, supplication, and burial. The non-natural phenomena resulting from Achilles’ slaughter do not incriminate him, but they reflect a disruption in the cosmic order, which is restored when Hephaestus burns the Scamander. In the Iliad, nature is a powerful force, but its presence is limited. Ultimately, the Iliad emphasises the need to domesticate nature in order to reduce its threat. In chapter 2, I looked at the mache parapotamios in Silius Italicus’ Punica. Many years after the Iliad’s lack of concern over the violation of the Scamander, the Punica presents an unambiguously moral argument against the mistreatment of nature. We saw that Silius thematises the destruction of nature in order to identify the virtuosity of individuals. This is most plain in the figure of Hannibal, but the mache parapotamios episode also questions the virtue of the elder Scipio, whose actions violate the riverine environment. Non-natural phenomena and other violations, such as Hannibal’s crossing of the Alps, are associated with boundary-crossing, which is presented as a moral failure. We also saw that the domestication of nature was of imperial significance, as Roman emperors sought to assert their claim to certain lands by co-opting aspects of the environment, especially rivers, to be their allies. 2 See Schmiel (2003). 74 In chapter 3, I discussed the mache parapotamios in Statius’ Thebaid and how the poet altered the Homeric source model in order for it to better suit a civil-war narrative. The unique focus on the pathos of the scene is an indicator of the brutality of the war, and the destruction of the environment is linked to the cosmic crisis brought about by civil strife. Moreover, we saw that there is ambiguity in Statius’ treatment of nature, which challenges previous ideas about domestication in particular. Like the Punica, the Thebaid uses the mistreatment of nature as one way to measure morality, and so Hippomedon’s assault on the river, and by extension his character, is presented as immoral. Although the theory with which I have proposed to study these scenes is by no means perfect, it is a useful tool for understanding the presentation and use of nature in epic. ἀασάµην, Διόνυσε πυριτρεφές: οὐρανίην γὰρ σῶν δαΐδων ἀµάρυγµα τεὴν κήρυξε γενέθλην. ἀλλὰ πόθος τεκέων µε βιήσατο (24.13-5). Through the domesticated/undomesticated binary, the epic poets are able to clarify some of the ambiguities surrounding nature’s place in civilisation, and explain the perceived unpredictable hostility and/or amiability of nature. By assigning human emotions and motives to the rivers, the authors manufacture reasons for which they might flood, change course, or remain tranquil. The volatility of the rivers also represents the volatility of nature more generally, and thus analysing the actions of rivers in epic also provides an insight into attitudes towards nature in epic overall. Throughout this thesis, I have shown that the presence of nature in these epics is not just a topographical necessity. It is clear in these examples, especially the Flavian texts, that nature is a formidable presence in the world, and that its various processes affect humans in significant ways. Conversely, the poems also acknowledge that human activities can influence the environment, sometimes to the detriment of human society. Overall, the epics emphasise that nature and civilisation are co- dependent, rather than in conflict, despite the threats that both pose to each other. They are also presented as separate: humankind is not envisaged as a part of nature as trees, mountains, rivers and non-human animals are. Indeed, the humans of the epics are styled as successful or unsuccessful domesticators of nature. Despite the usually positive connotations of domestication, at times it is presented with more moral ambiguity as questions arise over whether humans should have equal footing with, or power over, that which is divine or of divine origin. On the other hand, valuation of nature as a part of the world unto itself, irrespective of the divine, is not present, and there is no sense of responsibility for preserving nature as it is. Yet, despite a lack of 75 environmentalism, per se, Greco-Roman epic is nevertheless mindful of the centrality of nature, whether positive or negative, to human life. environmentalism, per se, Greco-Roman epic is nevertheless mindful of the centrality of nature, whether positive or negative, to human life. 76 Austin (1975) Archery at the Dark of the Moon: Poetic Problems in Homer’s Odyssey (Berkeley: University of California Press). 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W4220937226.txt_1	Open-Science-Pile	Open Science	Various open science	2,022	Active Peripheral Osteoma in the Mandibular Notch: A Longitudinal Clinical Report	None	English	Spoken	2,122	3,890	Hindawi Case Reports in Dentistry Volume 2022, Article ID 6364599, 5 pages https://doi.org/10.1155/2022/6364599 Case Report Active Peripheral Osteoma in the Mandibular Notch: A Longitudinal Clinical Report Jun Sasaki ,1,2 Shogo Hasegawa ,1 Hitoshi Miyachi,1 and Toru Nagao 1 1 Department of Maxillofacial Surgery, School of Dentistry, Aichi-Gakuin University, 2-11 Suemori-Dori, Chikusa, Nagoya, Aichi 464-8651, Japan 2 Department of Maxillofacial Surgery, Okazaki City Hospital, 3-1 Goshoai, Koryujicho, Okazaki, Aichi 444-8553, Japan Correspondence should be addressed to Shogo Hasegawa; shogo@dpc.aichi-gakuin.ac.jp Received 21 December 2021; Revised 12 February 2022; Accepted 10 March 2022; Published 22 March 2022 Academic Editor: Yasuhiro Morimoto Copyright © 2022 Jun Sasaki 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. Osteomas are benign osteogenic tumors composed of mature compact or cancellous bone and are characterized by slow, painless growth. A peripheral osteoma located in the mandibular notch is extremely rare. Here, we describe the case of a 38-year-old woman with an active peripheral osteoma in the mandibular notch for long-term follow up. Surgical resection of the lesion was performed, which resulted in the improvement of the mandibular function and temporomandibular dysfunction. This is the ﬁrst report of resection surgery after conﬁrming the activity of peripheral osteoma by bone scintigraphy. Creating a threedimensional model of the mandible promises an accurate surgical plan. 1. Introduction Osteomas are benign tumors composed of mature compact or cancellous bone and are characterized by slow, painless growth [1–3]. They can be peripheral, central, or extraskeletal [4, 5]. Most peripheral osteomas are known to occur in the mandibular body, angle, condyle, or coronoid process [2, 4, 5]. The occurrence of a peripheral osteoma in the mandibular notch is extremely rare, with an incidence of 1.6% among mandibular peripheral osteoma cases [4, 5]. Although several studies have described peripheral osteomas of the jawbone, only eight cases of peripheral osteomas in the Sigmoid notch of the mandible have been reported. We report the case of an active peripheral osteoma in the mandibular notch of a 38-year-old woman in whom we were able to clarify the temporal course of the disease. 2. Case Presentation A 38-year-old woman with complaints of pain during mandibular movement was referred to our department by a den- tal clinic. She had no signiﬁcant medical history, and she was not taking any medication. The patient was evaluated based on preoperative clinical manifestations, panoramic radiography, computed tomography (CT), magnetic resonance imaging (MRI), and radioisotope imaging (RI). Panoramic radiography revealed a circular radiopacity between the right mandibular condyle and the muscle process (Figure 1(d)). On reviewing the patient’s panoramic radiograph taken at the age of 18 and 21 years, no osteoma was found in the mandibular notch (Figures 1(a) and 1(b)). At the age of 26, a bone bulge was found in the right mandibular notch (Figure 1(c)). CT showed a 13.3×12.8×10.0mm lesion showing Hounsﬁeld units (HU) equivalent to that of the mandible, arising from the right mandibular notch (Figures 2(a) and 2(b)). MRI showed uniform hypointense trabecular bone, and there was no continuity of fat signals in the bone marrow, suggesting the exclusion of the lateral pterygoid muscle due to bone lesions (Figure 2(c)). RI showed signiﬁcant uptake of Tc-99m (Figure 2(d)). Bone scintigraphy showed circular area of strong radioisotope accumulation with indistinct boundaries, consistent with lesion in right mandibular notch. No neurological symptoms of the 2 Case Reports in Dentistry (a) (b) (c) (d) Figure 1: Time course of osteoma in the right mandibular notch by panoramic radiography. (a) At the age of 18, no osteoma was found in the right mandibular notch. (b) At the age of 21, no osteoma was found in the right mandibular notch. (c) At the age of 26, a bony bulge was found in the right mandibular notch. (d) At the age of 38, an oval radiodense mass was observed in the right mandibular notch. mandible were observed, and there was no history of trauma or infection in the maxillofacial region. No abnormalities were found on blood or urine analysis. Based on these ﬁndings, the diagnosis of an active peripheral osteoma was suspected. Using the CT data and Mimics software® (version 23.0, Materialise, Leuven, Belgium), we created a threedimensional (3D) model of the mandible (Figure 3) to facilitate an explanation to the patient. Resective surgery using an intraoral approach was planned. A fused deposition modeling 3D printer (da Vinci Mini w Plus, XYZ printing, Taiwan) and a thermoplastic resin (polylactic acid ﬁlament) were used to prepare the model. On June 23, 2021, the entire tumor, including the portion that demonstrated Tc-99m uptake, was resected under general anesthesia. The myoprojection was temporarily dissected (Figure 4(a)), and the bone was repositioned and ﬁxed with an absorbent plate (Super FIXSORB®) (Figure 4(b)). The excised mass measured 13×12.0×10.0mm with a weight of 2.7 g (Figure 4(c)). On histologic examination, osteomas are composed of dense, mature, predominantly lamellar bone. The osteoma was diagnosed an actively growing lesion (“hot”) with radionuclide bone imaging, interosseous spaces composed of ﬁbrous, ﬁbrovascular, and lined by osteoblasts within a well-vascularized moderately cellular ﬁbrous stroma (Figure 5). The case presented here was diagnosed as a compact osteoma because of its characteristic microscopical features, radiographic ﬁndings, and clinical features. Immediately after the operation, there was a slight trismus with no other complications. There were few functional changes and minimal temporomandibular joint dysfunction. The patient was followed-up for 5 months, and no recurrence was noted. Case Reports in Dentistry 3 (a) (b) (c) (d) Figure 2: Preoperative examination. (a) Computed tomography showed a 13.3×12.8×10.0mm sized lesion showing Hounsﬁeld units (HU) equivalent to that of the mandible, arising from the right mandibular notch. The CT values of osteoma ranged from 1480 to 1580 HU. (b) 3D-CT showed osteoma was found in the right mandibular notch. (c) Magnetic resonance imaging showed uniform hypointense trabecular bone, and there was no continuity of fat signals in the bone marrow, suggesting exclusion of the lateral pterygoid muscle because of bone lesions. (d) Radioisotope imaging showed signiﬁcant uptake of Tc-99m by the osteoma in the right mandibular notch. 3. Discussion Figure 3: Three-dimensional model of the mandible. We created a three-dimensional model of the mandible to facilitate an explanation to the patient resection surgery using an intraoral approach was planned. Peripheral osteomas, also known as periosteal osteomas, most frequently involve the craniofacial bones and are the most common benign tumors of the paranasal sinuses. Peripheral osteomas of the jawbones are however uncommon [2]. The origin of peripheral osteoma onset is unknown, and it has been suggested that it results from a reaction to trauma and infection [2]. However, there was no history of infection or trauma in the present case. The osteoma may have slowly developed during the age of 21– 26 years after the cessation of mandibular growth and development of secondary sexual characteristics. Surgery is considered for large, deforming osteomas [1, 6]. In the present case, surgical resection was considered owing to the progressive nature of the lesion conﬁrmed by RI. Only eight cases of osteomas in the mandibular notch have been reported so far [1, 6–10], and the present case is the ninth (Table 1). In the previous cases, the patients’ age ranged from 16 to 78 years, with a male-to-female ratio of 4 Case Reports in Dentistry (a) (b) (c) Figure 4: Intraoperative view. (a) The lesion was exposed by an intraoral approach. (b) The myoprojection was temporarily dissected, and the bone was repositioned and ﬁxed with an absorbent plate. (c) Excised osteoma. (a) (b) Figure 5: The histopathology of the peripheral osteoma. (a) Microscopic features of peripheral osteoma (hematoxylin-eosin, original magniﬁcation ×200). (b) Microscopic features of peripheral osteoma (hematoxylin-eosin, original magniﬁcation ×400). 1 : 3.5. The location of the osteoma was medial in four cases, lateral in four cases, and medial and lateral in one case. The lesion developed in the right mandibular notch in ﬁve cases and in the left mandibular notch in four cases. The patients displayed symptoms of swelling in four cases and pain in two, with no symptoms in three cases. Of Case Reports in Dentistry 5 Table 1: Reported cases of peripheral osteoma in the mandibular notch. No. 1 2 3 4 5 6 7 8 9 Reference Year of Age (years) publication Present case Nakayama et al. [3] Bocchialini et al. [1] Sasidharan et al. [6] Rao et al. [7] Iwai et al. [4] Sekerci et al. [8] Schulze [9] Bessho et al. [10] 2021 2021 2018 2015 2013 2013 2011 2008 1987 38 69 37 52 16 78 59 73 26 Sex Location Female Medial Female Lateral Female Medial and lateral Female Lateral Female Lateral Female Medial Male Medial Female Medial Male Lateral the ﬁve surgically treated cases of osteoma, four were intraoral approaches. This is the ﬁrst case to use RI to conﬁrm the dynamic state of the osteoma; RI is therefore useful for diagnosing osteomas. In addition, a 3D model was created based on the preoperative CT data using a fused deposition modeling 3D printer and a thermoplastic resin (polylactic acid ﬁlament). The 3D model is useful for preoperative patient education and surgical planning. In this case, because a 3D model was created only for the mandible, the surgery of the maxillary tuberosity and maxillary teeth was restricted, and it was diﬃcult to predict. Among the reports of osteomas in the mandibular notch, there were descriptions of intraoral surgery in four cases. In the present case, considering the patient’s aesthetics, we approached the lesion intraorally. Currently, the number of reported cases of peripheral osteoma in the mandibular notch is small, and many aspects of this entity remain unclear, including the treatment method and prognosis. Further accumulation and examination of cases are expected. 4. Conclusions We encountered a case of an active peripheral osteoma in the mandibular notch, which has rarely been reported in the literature. A dynamic diagnosis of peripheral osteoma was made possible using composite imaging. It was easy to obtain anatomical orientation in the surgical ﬁeld through preoperative surgical simulation using a 3D model. An osteoma in the mandibular notch can be resected by an intraoral approach, in which case it is desirable to reposition the myoprojection. Consent The ﬁgures used are completely anonymized. The patient’s permission was obtained. Conflicts of Interest The authors have no conﬂict of interest to declare. Side Size (in mm) Symptoms Right 15×14×10 Left 25×20×15 Left 15×13×11 Left 30×25×20 Right 30×30 Right 36×35×30 Right Not available Left 30×30×27 Right 34×29×17 Pain Swelling Pain Swelling Swelling None None Pain Swelling Treatment Surgery (intraoral) Surgery (extraoral) Surgery (intraoral) Surgery (intraoral) Surgery (intraoral) Follow-up Follow-up Unknown Surgery References [1] G. Bocchialini, A. B. Bolzoni Villaret, S. Negrini, A. Tironi, L. Salvagni, and A. Castellani, “The ﬁrst case of osteoma of the mandibular notch located both medially and laterally and treated with a transoral endoscopy assisted approach. A case report,” International Journal of Surgery Case Reports, vol. 49, pp. 70–73, 2018. [2] I. Kaplan, S. Calderon, and A. Buchner, “Peripheral osteoma of the mandible: a study of 10 new cases and analysis of the literature,” Journal of Oral and Maxillofacial Surgery, vol. 52, no. 5, pp. 467–470, 1994. [3] A. Nakayama, R. Abe, M. Takahashi, H. Yamada, and Y. Takeda, “Peripheral osteoma in an unusual region of the mandible: a case report,” Oral and Maxillofacial Surgery Cases, vol. 7, no. 1, article 100207, 2021. [4] T. Iwai, T. Izumi, J. Baba, J. Maegawa, K. Mitsudo, and I. Tohnai, “Peripheral osteoma of the mandibular notch: report of a case,” Iranian Journal of Radiology, vol. 10, no. 2, pp. 74–76, 2013. [5] C. T. Chen and K. Adriane, “Endoscopic resection of a mandibular body and condylar osteoma,” Minimally Invasive Therapy & Allied Technologies, vol. 17, no. 5, pp. 323–325, 2008. [6] A. Sasidharan, A. Gohil, S. Koshy, and A. K. Gupta, “Bony tumour in an unusual location on the mandible,” Indian Journal of Plastic Surgery, vol. 48, no. 2, pp. 200–203, 2015. [7] S. Rao, S. Rao, and D. S. R. Pramod, “Transoral removal of peripheral osteoma at sigmoid notch of the mandible,” Journal of Maxillofacial and Oral Surgery, vol. 14, Suppl. 1, pp. 255– 257, 2015. [8] A. E. Sekerci, H. Sahman, E. T. Ertas, and Y. Sisman, “An unusual peripheral osteoma of the mandibular notch: a case report with computed tomography evaluation,” Oral Radiology, vol. 27, no. 1, pp. 78–82, 2011. [9] D. Schulze, “Radiographic diagnostics: osteoma of the left mandibular notch,” Quintessence International, vol. 39, no. 8, pp. 702-703, 2008. [10] K. Bessho, K. Murakami, T. Iizuka, and T. Ono, “Osteoma in mandibular condyle,” International Journal of Oral and Maxillofacial Surgery, vol. 16, no. 3, pp. 372–375, 1987.	17,746
https://openalex.org/W2504215566	OpenAlex	Open Science	CC-By	2,016	Generation and Characterization of an IgG4 Monomeric Fc Platform	Shijie Lu	English	Spoken	10,777	19,249	RESEARCH ARTICLE Generation and Characterization of an IgG4 Monomeric Fc Platform Lu Shan1, Magali Colazet1, Kim L. Rosenthal1, Xiang-Qing Yu2, Jared S. Bee3, Andrew Ferguson4, Melissa M. Damschroder1, Herren Wu1, William F. Dall’Acqua1, Ping Tsui1, Vaheh Oganesyan1 1 Antibody Discovery and Protein Engineering, MedImmune, Gaithersburg, Maryland, United States of America, 2 Clinical Pharmacology and Drug Metabolism and Pharmacokinetics, MedImmune, Gaithersburg, Maryland, United States of America, 3 Analytical Biotechnology, MedImmune, Gaithersburg, Maryland, United States of America, 4 Discovery Sciences, Structure and Biophysics, AstraZeneca Pharmaceuticals, Waltham, Massachusetts, United States of America a1111 * TsuiP@medimmune.com (PT); OganesyanV@MedImmune.com (OV) Data Availability Statement: Structural data were uploaded to Protein Data Bank under Id. 5HVW. Data Availability Statement: Structural data were uploaded to Protein Data Bank under Id. 5HVW. Abstract The immunoglobulin Fc region is a homodimer consisted of two sets of CH2 and CH3 domains and has been exploited to generate two-arm protein fusions with high expression yields, simplified purification processes and extended serum half-life. However, attempts to generate one-arm fusion proteins with monomeric Fc, with one set of CH2 and CH3 domains, are often plagued with challenges such as weakened binding to FcRn or partial monomer formation. Here, we demonstrate the generation of a stable IgG4 Fc monomer with a unique combination of mutations at the CH3-CH3 interface using rational design com- bined with in vitro evolution methodologies. In addition to size-exclusion chromatography and analytical ultracentrifugation, we used multi-angle light scattering (MALS) to show that the engineered Fc monomer exhibits excellent monodispersity. Furthermore, crystal struc- ture analysis (PDB ID: 5HVW) reveals monomeric properties supported by disrupted inter- actions at the CH3-CH3 interface. Monomeric Fc fusions with Fab or scFv achieved FcRn binding and serum half-life comparable to wildtype IgG. These results demonstrate that this monomeric IgG4 Fc is a promising therapeutic platform to extend the serum half-life of pro- teins in a monovalent format. * TsuiP@medimmune.com (PT); OganesyanV@MedImmune.com (OV) Editor: Sergey Korolev, Saint Louis University, UNITED STATES Editor: Sergey Korolev, Saint Louis University, UNITED STATES Received: March 4, 2016 Accepted: July 18, 2016 Published: August 1, 2016 Copyright: © 2016 Shan 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. Copyright: © 2016 Shan 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. OPEN ACCESS OPEN ACCESS Citation: Shan L, Colazet M, Rosenthal KL, Yu X-Q, Bee JS, Ferguson A, et al. (2016) Generation and Characterization of an IgG4 Monomeric Fc Platform. PLoS ONE 11(8): e0160345. doi:10.1371/journal. pone.0160345 Editor: Sergey Korolev, Saint Louis University, UNITED STATES Received: March 4, 2016 Accepted: July 18, 2016 Published: August 1, 2016 Copyright: © 2016 Shan 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. Citation: Shan L, Colazet M, Rosenthal KL, Yu X-Q, Bee JS, Ferguson A, et al. (2016) Generation and Characterization of an IgG4 Monomeric Fc Platform. PLoS ONE 11(8): e0160345. doi:10.1371/journal. pone.0160345 Editor: Sergey Korolev, Saint Louis University, UNITED STATES IgG4 Monomeric Fc with Desirable Structure and Activity monovalent targeting to avoid receptor agonism caused by receptor dimerization from bivalent antibodies or Fc fusions [6]. Competing Interests: The authors have no competing interests. The commercial affiliation with MedImmune and AstraZeneca does not alter the authors' adherence to all PLOS ONE policies on sharing data and materials. Monovalent versions of Fc fusion proteins (Alprolix—coagulation factor IX fusion, Eloctate —factor VIII fusion) or monovalent antibodies (Onartuzumab—anti-cMet one-arm mAb) that have advanced to the clinic utilize an Fc domain that is engineered to form a heterodimer, either with tethering or “knobs-into-holes” technology [7, 9]. These, along with other heterodi- meric Fc technologies, rely on robust purification processes to remove undesired chain pairing and achieve a homogeneous fusion protein [10] (Fig 1A). To search for an alternative approach aimed at simplifying product development, there has been extensive effort in engineering fusion protein platforms with a monomeric Fc modality consisted of only one set of CH2 and CH3 domains (Fig 1B), either through weakening the interactions or by generating steric hin- drances with the addition of glycans at the CH3-CH3 dimer interface in the Fc [11–13]. So far these approaches have encountered challenges in several aspects, including solubility and sta- bility, loss of FcRn binding, or lack of homogeneity. Additionally, many of the previously engi- neered monomeric Fc molecules were observed by dynamic light scattering to have a tendency for aggregation, highlighting the challenge of stabilizing the monomeric conformation after weakening the homodimer interface [12, 14]. To date, the only available crystal structure of monomeric Fc has been the glycoengineered Fc monomer, where an additional glycan at the dimer interface resulted in a stable monomer [11]. There has also been some evidence that avidity of the bivalent Fc has a large contribution to FcRn binding [11]. This suggests that monomeric Fc, without additional half-life extension technology, would result in dramatic loss of binding to FcRn [11, 12]. To compensate for the lower FcRn binding affinity, linking mono- meric Fc in tandem format has been utilized [11], which further complicates the biophysical characteristics of the final fusion molecules. Abbreviations: Fc, fragment crystallizable; Fab, fragment antigen binding; scFv, single chain fragment variable; IgG, Immunoglobulin G; mAbs, monoclonal antibodies; CH3, heavy chain constant domain 3; SEC-MALS, size-exclusion chromatography coupled with multi-angle light scattering; PK, pharmacokinetics; hFcRn, human neonatal Fc receptor; FcγR, receptor for IgG Fc; YTE, M252Y/ S254T/T256E. Introduction The homodimeric immunoglobulin fragment crystallizable, Fc, has been widely utilized to form fusion proteins with enzymes, growth factors, immune modulators, and target-binding moieties such as scFv [1–4] (Fig 1A). Both as research tools and as therapeutic agents, Fc- fusion proteins are able to harness FcRn-mediated serum half-life extension provided by the Fc domain. In recent years, there have been several examples of proteins fused on one arm of the Fc, e.g., erythropoietin, coagulation factor IX, and interferon, that exhibited similar or improved stability and biological activities compared to conventional Fc fusions [5–8]. In addi- tion, there are particular signaling pathways, such as receptor tyrosine kinases, which require Funding: This study was funded solely by MedImmune which is a subsidiary of AstraZeneca Plc. AstraZeneca has provided support in the form of salaries for all of the authors [LS, MC, KLR, XQY, JSB, AF, MMD, HW, WFD, PT, VO], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the “Author Contributions” section. 1 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 Reagents Restriction enzymes and polymerases were purchased from New England Biolabs (Ipswich, MA) and Clontech (Mountain View, CA). Oligonucleotides were purchased from Integrated DNA Technologies (Coralville, Iowa). Recombinant human and murine FcRn were expressed and purified as described previously [15]. Antibody positions are listed according to the Kabat numbering convention [16]. Ethics statement Protocol (MI-13-0012) requiring the use of animals in these studies was reviewed and approved by MedImmune’s Institutional Animal Care and Use Committee and complies with the animal welfare standards of the USDA, Guide for the Care and Use of Laboratory Animals, and AAALAC international. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 We report here the development of a therapeutic platform for the expression of a mono- meric Fc fusion protein that exhibits FcRn binding affinity comparable to the wildtype Fc. We Fig 1. Cartoon representations of wildtype IgG Fc, monomeric Fc and fusion proteins. (A) Cartoons of Fc homodimer in IgG and in a bivalent Fc fusion protein, as well as a one-arm mAb and a monovalent Fc fusion, supported by heterodimeric Fc (as shown) or tethered Fc. (B) Cartoons of a monomeric Fc, along with Fab- and scFv- fusion proteins with a monomeric Fc. doi:10.1371/journal.pone.0160345.g001 Fig 1. Cartoon representations of wildtype IgG Fc, monomeric Fc and fusion proteins. (A) Cartoons of Fc homodimer in IgG and in a bivalent Fc fusion protein, as well as a one-arm mAb and a monovalent Fc fusion, supported by heterodimeric Fc (as shown) or tethered Fc. (B) Cartoons of a monomeric Fc, along with Fab- and scFv- fusion proteins with a monomeric Fc. Fig 1. Cartoon representations of wildtype IgG Fc, monomeric Fc and fusion proteins. (A) Cartoons of Fc homodimer in IgG and in a bivalent Fc fusion protein, as well as a one-arm mAb and a monovalent Fc fusion, supported by heterodimeric Fc (as shown) or tethered Fc. (B) Cartoons of a monomeric Fc, along with Fab- and scFv- fusion proteins with a monomeric Fc. doi:10.1371/journal.pone.0160345.g001 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 2 / 18 IgG4 Monomeric Fc with Desirable Structure and Activity devised a comprehensive protein engineering approach that involved using a unique IgG4 phage library design and thermal stability and folding selections, in addition to a pH-depen- dent FcRn binding selection, to identify a monomeric Fc with excellent monodispersity. Our results show that a library selection strategy combining thermal selection and rational template designs can lead to monomeric Fc fusion proteins which have the desired biophysical, struc- tural and pharmacokinetics (PK) properties. Phage library design, selection and screening An IgG4 Fc template containing M252Y/S254T/T256E (YTE) and F405R was constructed in a phage vector [15]. A phage library was built with random mutations at five additional CH3-CH3 interface residue positions, L351, S354, T366, P395 and Y407 [13, 17, 18]. Based on previous work showing Fc had favored monomer formation when mutated to R, Q or E from F at position 405, we fixed these three mutations in the library build [12, 13, 19]. Five primers were designed to cover the positions, L351/S354, T366, P395, Y407/F405RQ, Y407/F405E, respectively, to yield 3 overlapping PCR fragments, which were then annealed together. The library insert was ligated into the phage vector. The library diversity was achieved at 1×109 cov- ering all possible mutations. Human FcRn was expressed in human embryonic kidney cell line HEK293F(Life Technolo- gies, Grand Island, NY) and purified as a soluble protein [15]. Library selection by biopanning against FcRn generally followed previously described procedures [15], with some modifica- tions. Library phages were first passed through a Protein G column (Roche) to eliminate phages displaying misfolded Fc mutants which had lost Protein G binding [13]. For each round of the pH-dependent FcRn binding selection, amplified libraries of 1012 phages were biopanned against biotinylated FcRn. The libraries and streptavidin-coated magnetic Dyna- Beads (Thermo Fisher Scientific, Waltham, MA) were first blocked with 20% bovine serum albumin (BSA) at pH 6.0 for 1 hour. A deselection step was then carried out by mixing blocked beads and the libraries to remove non-specific bead-binding phages. Then the deselected libraries were incubated with 75 nM of biotinylated FcRn and the blocked beads at pH 6.0 for 2 hours. Extensive washes were carried out on KingFisher instrument (Thermo Fisher). The phages were eluted with phosphate buffered saline (PBS, 5 mM sodium phosphate and 155mM NaCl), 0.01% Tween-20, at pH 7.4. The eluted phages were grown and amplified using Escheri- chia coli TG1 cells (Lucigen, Middleton, MI) and prepared for the next round of selection. Starting at round 3 of library selection, library selection steps were performed with and without 3 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity thermal stress. Thermal stress was carried out by elevating the phage incubation temperatures from 37 to 50°C in rounds 3 and 4, and further increased to 55°C in round 5 [20]. Cloning, expression and purification of anti-cMet monomeric Fc fusion proteins Four FcRn binding clones identified from the library selection were used for mammalian expression. They were purified using Protein A column and further characterized. After con- firming its monomeric state, clone 4 (C4) was used to generate monomeric Fc fusion proteins with the Fab (Onart-Fab-C4) and the scFv (Onart-scFv-C4) of the antibody Onartuzumab [6]. Phage library design, selection and screening Forty eight clones were picked from rounds 4 and 5 of the library selection output and were expressed by inducing with 1 mM IPTG (isopropyl-1-thio-β-D-galactopyranoside, Sigma-Aldrich, St. Louis, MO) for 3 hours at 37°C. The bacterial lysates were analyzed by Western blotting under non- reducing conditions. Phages from each clone were prepared by infecting TG1 cells with helper phage MK13KO7 (Life Technologies) to be screened by FcRn binding ELISA at pH 6.0 and pH 7.4. Neutravidin at 5 μg/mL in PBS (pH 6.0) was coated on a Nunc Maxisorp Plate wells (Thermo Fisher) overnight at 4°C. After three washes, biotinylated FcRn was added at 2 μg/ mL, and the wells were blocked with 4% BSA in PBS buffer with 0.1% Tween 20. Phages diluted in 1% BSA in the same buffer were added and incubated for 1 hour. The plates were then washed three times and anti-M13-HRP antibody (GE Healthcare, Pittsburgh, PA) incubated for 1 hour. Binding was detected with the addition of SureBlue 3,30,5,50-tetramethylbenzidine (TMB) substrate (KPL, Gaithersburg, MD), and the reaction was stopped by adding the TMB Stop Solution (KPL). Absorbance signals were read at 450 nm. Phages showing positive bind- ing to FcRn in ELISA were sequenced and subsequently cloned into a proprietary mammalian vector for expression and purification. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 Size-exclusion chromatography with light scattering and analytical ultracentrifugation Purified Fc clones and fusion proteins at 1 mg/mL or higher were analyzed by size exclusion chromatography using a TSK-GEL G2000SWXL column with a 14 mL bed volume (Tosoh Biosciences, Tokyo, Japan) on an Agilent 1100 HPLC (Agilent, Santa Clara, CA) at room tem- perature. The samples were eluted isocratically in PBS at a flow rate of 1 mL/min for 20 min. Eluted proteins were detected using UV absorbance at a wavelength of 280 nm. Data analysis was done using the Agilent software ChemStation (version A.02.10). Column calibration was performed with a set of molecular weight standards ranging from 10 to 500 kDa (Bio-Rad, Her- cules, CA). In-line multi-angle light scattering (SEC-MALS) was performed. Sample measure- ments were done on a DAWN HELEOS II MALS with an Optilab Rex refractometer (Wyatt Technologies, Santa Barbara, CA). Molecular mass of each protein within a defined chro- matographic peak was calculated using ASTRA, version 6.1 (Wyatt Technologies). For analyti- cal ultracentrifugation analysis, samples and reference buffer were loaded into 12 mm double- sector cells with Epon centerpieces then placed in an An-50 rotor for ultracentrifugation at 42,000 rpm using a Beckman Optima XL-I centrifuge set to 20°C (Beckman-Coulter, Indianap- olis, IN). The sedimentation data collected at 280 nm for scans 2 to 140 were analyzed using the Sedfit software program (version 14.6e) to generate c(s) distributions [21, 22]. The partial molar volume was set to a value of 0.73 g/mL. Solution density and viscosity values for PBS were set to 1.00523 g/mL and 1.019 mPas, respectively, using the calculated value from the Sednterp program (version 20130813) [23]. Based on the Svedberg equation a monomeric Fc with molecular mass of 28 kDa is expected to have a sedimentation coefficient of between 1.8 to 2.6 S (Svedbergs) assuming a frictional ratio of 1.3 to 1.8 (globular to extended shape). PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 4 / 18 IgG4 Monomeric Fc with Desirable Structure and Activity Crystallization, data collection and structure determination Prior to crystallization, Protein A-purified C4n (clone C4 with no YTE) was further purified by ion exchange chromatography on a Q HP 5 mL prepacked column (GE) equilibrated with 25 mM HEPES buffer at pH 7.5, followed by size exclusion chromatography using a Sepharose 75 column (GE) pre-equilibrated with 25 mM HEPES buffer with 100 mM NaCl at pH 7.5. Isocra- tically eluted protein was concentrated to 4 mg/mL and subjected to crystallization using com- mercial crystallization screens in a sitting drop format with Phoenix (ARI, Mountain View, CA). Diffraction quality crystals were grown in the crystallization optimization step in a hang- ing drop format from a solution containing 9 mM ZnCl2, 90 mM HEPES, 18% PEG 6000 (w/v) at pH 7.0. Cryoprotection was achieved by transferring crystals through several solutions of increasing glycerol concentrations, up to 20% v/v, added to the crystal growth solution. Crys- tals were flash cooled by dipping in liquid nitrogen. Complete diffraction data to 1.95 Å were collected on LS-CAT beamline 21-ID at Argonne National Laboratory (Lamont, IL) using x- ray wavelength 1.00Å and oscillation range of 0.5°. Seven hundred twenty images were col- lected using Mar300 CCD detector and processed using HKL2000 [24]. All crystallographic calculations were carried out using the CCP4 software suite (version 6.5) [25]. The molecular replacement procedure was carried out using Molrep [26]. Structure refinement was performed using Refmac5 and model adjustments were carried out using the “O” program [27, 28]. Images for the structure figures were generated using PyMOL (Schrödinger, LLC). Octet binding analysis Binding measurements of the monomeric Fc and its fusion proteins to in-house purified recombinant human and murine FcRn, FcγRI, FcγRIIIa and an inactive form of IgG-degrading enzyme of S. pyogenes (IdeS, C94S) [29] were carried out by Bio-layer Interferometry on an Octet384 instrument (ForteBio, Menlo Park, CA). Biotinylated FcRn at 1 μg/mL in PBS buffer (pH 7.4) or 100 mM MES buffer (pH 6.0), with 3 mg/ml BSA, 0.05% (v/v) Tween 20 (1× Kinet- ics Buffer, ForteBio) were captured on streptavidin (SA) biosensors (ForteBio). The loaded bio- sensors were washed with assay buffer to remove any unbound protein, followed by association and dissociation measurements with serial dilutions of the different Fc variants or Fc-fusion constructs. Kinetic parameters (kon and koff) and apparent affinities (KD) were calculated from a non-linear fit based on the 1:1 binding kinetic model ðA þ B$ kon koff ABÞ of the data using the Octet384 software (v.7.2). Kinetic measurements of anti-cMet fusion proteins to recombinant human cMet protein were also performed. Fc-fusion proteins were captured on anti-human Fc (AHC) biosensors (ForteBio) in PBS buffer, pH 7.2, with 1× kinetics buffer. Binding steps were done with serial dilutions of cMet. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 cMet activity assays Cell viability was measured by the CellTiter-Glo1 Luminescent Cell Viability Assay (Promega, Madison, WI) kit. Lovo cells were trypsinized, washed and seeded at 100 μl in 96-well plates at a density of 1 × 104 cells/well in serum-free cell media with 0.1% BSA. After 2-hour incubation at 37°C in 5% CO2 atmosphere, 50 μl of antibodies at various concentrations in triplicates were added to the cells and incubated for another hour. Then 50 μl of human growth factor (HGF) was added to the ligand-containing wells at the final concentration of 20 ng/mL. The plates were incubated for 72 h at 37°C. After ligand treatment, the cells were exposed to the CellTiter- Glo1 reagent for 20 min and the luminescence was measured using an EnVision plate reader (PerkinElmer, Waltham, MA). The IC50 values were defined as the antibody concentrations inhibiting cell growth by 50%. 5 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity PK data analysis Non-compartmental PK data analysis was performed using Phoenix 64 WinNonlin 6.3 (Phar- sight, Mountain View, CA). The maximum observed peak plasma concentration (Cmax) was determined by inspection of the observed data using WinNonlin. The terminal elimination half-life (t½) was determined using the equation ln(2)/λz, where λz is the slope of the terminal portion of the natural-log concentration-time curve, determined by linear regression of at least the last 3 time points. The systemic exposure was determined by calculating the area under the curve for the plasma concentration versus time graph (AUClast) from the start of dosing to the time of last measurable concentration using the linear/log trapezoidal rule. Area under the curve for the plasma-concentration vs. time graph from time 0 to infinity (AUC1) was calcu- lated as: AUClast + Clast/λz, where Clast is the last quantifiable concentration. Clearance (CL) was calculated by dose/AUC1, and steady state volume of distribution (Vss) was calculated as: (AUMC1×CL)/AUC1, where AUMC1 is the area under the curve from the first moment extrapolated to infinity. PK parameters were summarized statistically and presented as mean and one standard deviation. In vivo PK studies in hFcRn transgenic mice hFcRn transgenic mice used in this study are the F1 cross of murine FcRn-deficient B6.129X1- Fcgrttm1Dcr/DcrJ and hFcRn cDNA transgenic line B6.Cg-Fcgrttm1Dcr Tg (CAG-FCGRT) 276 Dcr/DcrJ [30, 31]. Sex-matched (6–16-week-old) mice were given a bolus intravenous dose of 2.5 mg/kg monomeric Fc fusion proteins on day 0. Eight mice were used per protein, with two groups of mice (A group or B group) bled at alternate time points. Blood samples were obtained from the retro-orbital plexus using capillary pipettes at different time points through- out the 2–3-week long study. All animals remained healthy throughout the study. A quantita- tive ELISA was used to monitor the serum concentrations of the tested antibodies. Briefly, 96-well plates were coated with 2 μg/ml cMet-ECD (extracellular domain). Plates were blocked with 3% BSA in PBS at pH 7.2 for 1 hour and then incubated with appropriately diluted serum samples. Goat anti-human Fc-specific HRP-conjugated antibody (dilution 1:5000; Jackson ImmunoResearch Laboratories, West Grove, PA) was used for detection. Absorbance at 450 nm was measured after development with 3,30,5,50-tetramethylbenzidine substrate (KPL) according to the manufacturer's directions. Standard curves were generated for each antibody variant diluted into 1:100 prebleed mouse sera taken at day −3. The linear portions of standard curves generated in Prism (Version 6, GraphPad Software, La Jolla, CA) were then used to quantify human anti-cMet fusion proteins in the serum samples. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 Design and screening for monomeric IgG4 Fc The Fc homodimer interface is a well packed region between two CH3 domains mediated by over 16 residues in all of the IgG subclasses [13, 17, 18]. In particular, IgG4 is unique in its abil- ity to form monomer-dimer equilibrium, partially due to the contribution of the K409R point mutation compared to IgG1-3 [32, 33]. Previous work with scanning mutagenesis gave valu- able insight for key interface residues in IgG4 that contributed to the homodimer formation [12]. Based on these findings, we designed a phage library to maximize the chance to identify an IgG4 monomeric Fc with improved FcRn binding affinity, homogeneity and stability (Fig 1B), by fully exploring the sequence combinations at the CH3-CH3 interface positions, L351, S354, T366, P395 and Y407, with random mutagenesis. We fixed point mutations F405R/E/Q in the library build, which had been shown previously to be favored in a monomeric state [12, 6 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity 13, 19]. In addition, YTE mutations (M252Y/S254T/T256E), known for IgG half-life extension via tighter FcRn binding, were placed in the library template, so that any selected final sequence will be compatible with these residues (Fig 2A). 13, 19]. In addition, YTE mutations (M252Y/S254T/T256E), known for IgG half-life extension via tighter FcRn binding, were placed in the library template, so that any selected final sequence will be compatible with these residues (Fig 2A). To increase the selection pressure on more stable monomeric clones, we incorporated a pro- tein G binding selection and a thermal challenge to the library phages, in addition to the pH- dependent FcRn binding selection (Fig 2A) [20]. The phages were incubated at 50°C during Fig 2. Monomeric Fc library design, selection workflow and output analysis. (A) Phage display library with mutations in CH2 and CH3 was built as indicated in the sequence alignment. X represents any amino acid and Z represents R, E or Q. The library selection was carried out first with Protein G binding, then several iterative rounds of thermal stress and FcRn binding. At the end of the library selection, randomly picked clones were sequenced and the frequency of amino acids at each position, in single letter codes, was plotted in the bar graphs. Parallel biopanning rounds with no thermal stress (B) and with thermal stress (C) showed the improved enrichment under thermal selection pressure. doi:10.1371/journal.pone.0160345.g002 Design and screening for monomeric IgG4 Fc doi:10.1371/journal.pone.0160345.g002 To increase the selection pressure on more stable monomeric clones, we incorporated a pro- tein G binding selection and a thermal challenge to the library phages, in addition to the pH- dependent FcRn binding selection (Fig 2A) [20]. The phages were incubated at 50°C during Fig 2. Monomeric Fc library design, selection workflow and output analysis. (A) Phage display library with mutations in CH2 and CH3 was built as indicated in the sequence alignment. X represents any amino acid and Z represents R, E or Q. The library selection was carried out first with Protein G binding, then several iterative rounds of thermal stress and FcRn binding. At the end of the library selection, randomly picked clones were sequenced and the frequency of amino acids at each position, in single letter codes, was plotted in the bar graphs. Parallel biopanning rounds with no thermal stress (B) and with thermal stress (C) showed the improved enrichment under thermal selection pressure. 7 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity Fig 3. Analytical size-exclusion chromatograms of purified lead Fc clones 1, 2, 4 and 6. The retention times of clones 1, 2 and 6 appear shorter than that of clone 4, suggesting a difference in their molecular masses. Fig 3. Analytical size-exclusion chromatograms of purified lead Fc clones 1, 2, 4 and 6. The retention times of clones 1, 2 and 6 appear shorter than that of clone 4, suggesting a difference in their molecular masses Fig 3. Analytical size-exclusion chromatograms of purified lead Fc clones 1, 2, 4 and 6. The retention times of clones 1, 2 and 6 appear shorter than that of clone 4, suggesting a difference in their molecular masses. Fig 3. Analytical size-exclusion chromatograms of purified lead Fc clones 1, 2, 4 and 6. The retention times of clones 1, 2 and 6 appear shorter than that of clone 4, suggesting a difference in their molecular masses. doi:10.1371/journal.pone.0160345.g003 rounds 3 and 4 of biopanning, and at 55°C in round 5. Whereas very little amino acid enrich- ment was seen in the biopanning rounds against FcRn binding without thermal selection, the biopanning done with thermal selection was able to achieve notable enrichment (Fig 2B and 2C). For position 351, the mutations narrowed down to residues L, I, F, T and V. Design and screening for monomeric IgG4 Fc Position 354 was enriched to residues S, T and L. Position 366 has the most enrichment to W, followed by G, R and S. Position 395 is preferentially selected to more polar residues (C, E and K). The three F405 mutations to Q, E and R in the library template were seen to have narrowed down R and Q. Tyrosine residue at position 407 is changed to V and L, and to a smaller extent, polar residues (E and H). doi:10.1371/journal.pone.0160345.t001 Characterization of library-derived clones Clones derived from the thermal selection were screened for FcRn binding using phage ELISA. Four FcRn binding clones 1, 2, 4 and 6 were selected to express corresponding proteins using mammalian cells. Purified proteins were analyzed by SEC-MALS. All the mutants showed good homogeneity with clones 1, 2 and 6 showing slightly shorter retention times in the SEC profiles (Fig 3). The MALS data revealed significantly different molecular weights for these clones (Table 1). Clone 4 (C4) showed the expected molecular mass of ~27 kDa, confirming its monomeric state, which was further confirmed by analytical ultracentrifugation, with a Table 1. Molecular weights of Fc fragments and fusion proteins determined by MALS. Construct Mn (g/mol) Mp (g/mol) Mw (g/mol) Mw/Mn Clone 4 2.7×104 (±0.8%) 2.7×104 (±0.8%) 2.7×104 (±0.8%) 1.001 (±1.2%) Clone 1 5.7×104 (±0.3%) 5.7×104 (±0.2%) 5.7×104 (±0.3%) 1.000 (±0.4%) Clone 2 5.3×104 (±0.3%) 5.3×104 (±0.1%) 5.3×104 (±0.3%) 1.001 (±0.4%) Clone 6 5.3×104 (±0.2%) 5.4×104 (±0.1%) 5.3×104 (±0.2%) 1.000 (±0.3%) Onart-Fab-C4 7.7×104 (±0.9%) 7.6×104 (±0.9%) 7.7×104 (±0.9%) 1.000 (±1.2%) Onart-scFv-C4 5.9×104 (±0.9%) 5.8×104 (±0.9%) 5.9×104 (±0.9%) 1.001 (±1.3%) Mn represents number-average molar mass. Mp represents molar mass at concentration peak. Mw represents weight-average molar mass. Mw/Mn represents polydispersity. Table 1. Molecular weights of Fc fragments and fusion proteins determined by MALS. Mn represents number-average molar mass. Mp represents molar mass at concentration peak. Mw represents weight-average molar mass. Mw/Mn represents polydispersity. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 8 / 18 IgG4 Monomeric Fc with Desirable Structure and Activity measured sedimentation coefficient of 2.2 S (expected monomer sedimentation coefficient is between 1.8 and 2.6 S). Analysis of C4 and C4n (C4 without YTE) at high concentrations (>10 mg/mL) by SEC found that the proteins remained monomeric (data not shown). Aside from the YTE mutations, the mutations at the CH3-CH3 dimer interface in C4 were L351F, T366R, P395K, F405R, and Y407E. Interestingly, clones 1, 2 and 6, which did not show mono- meric characteristics in SEC and MALS, had common mutations L351I, T366W and Y407L, possibly resulting in increased hydrophobicity at the interface. Only C4 was carried forward as a monomeric Fc to generate fusion proteins to achieve better FcRn binding characteristics with the YTE mutations, while C4n was used to determine the crystal structure to present the mono- meric Fc in a more general way (without YTE). PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 Structure determination and refinement The monomeric Fc mutant C4n formed sizeable rhombohedral shape crystals that diffracted to a resolution of 1.95 Å. The morphology of crystals corresponded to their symmetry R32 (SG #155). In a hexagonal setting (H32) the cell parameters are a = b = 121.4 Å, c = 99.7 Å. The asymmetric part of the unit cell contained one polypeptide with Matthew’s parameter of 2.7 Da/Å3 with a solvent content of 55%. The structure was solved by molecular replacement. As a template for molecular replacement, a 1.9 Å resolution IgG4 Fc model from PDB ID 4C54 was used [34]. For most of the molecule, the initial electron density maps were of excellent quality, allowing for the placement of correct amino acids at the mutation sites. The X-ray data and refinement statistics are shown in Table 2. The root-mean-square deviation (RMSD) of Cα atom coordinates between the final struc- ture and the previously solved IgG4 Fc structure is only 0.53Å. This high degree of similarity renders these molecules as nearly identical. However, the CH3-CH3 interface of a conventional Fc homodimer was not observed. The conformation of the last 15 amino acids of in the Table 2. X-ray data and model refinement statistics. Data statistics Wavelength (Å) 1.0000 Resolution (Å) 46.50–1.95 (2.02–1.95)* Space group H32 Unit-cell parameters (Å) a = 121.4, b = 121.4, c = 99.7 Total reﬂections 229813 (20424) Unique reﬂections 19638 (2038) Completeness (%) 99.9 (100.0) Rmerge 0.064 (0.718) Mean I/σ (I) 33.1 (2.2) Multiplicity 11.1 (11.3) Reﬁnement statistics Resolution (Å) 24.00–1.95 Rwork/Rfree/Rwork+free 0.199/0.259/0.202 RMSD bonds (Å) 0.018 RMSD angles (°) 2.041 Ramachandran outliers (%) 0.0 Number of protein atoms 1679 Number of non-protein atoms 199 Mean B factor (Model/Wilson) (Å2) 47.5/50.0 Values in parentheses correspond to the highest-resolution shell doi:10.1371/journal.pone.0160345.t002 Table 2. X-ray data and model refinement statistics. Table 2. X-ray data and model refinement statistics. PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 9 / 18 IgG4 Monomeric Fc with Desirable Structure and Activity monomeric Fc structure deviates substantially from the wildtype Fc. This phenomenon is due to the presence of Zn2+ ions in the crystallization solution (S1 Fig). The final model contains 1679 protein atoms, 110 carbohydrate atoms, two Zn ions, one glycerol molecule and 81 sol- vent molecules (Fig 4A). Monomeric Fc binding to Fc receptors We compared apparent binding affinities of C4 and C4n, along with Fc controls, to human FcRn. As expected, C4 and C4n showed no measurable binding to FcRn at pH 7.4 (data not shown). Using a steady-state binding analysis, we determined that at pH 6.0, C4 demonstrated similar FcRn binding (370 nM) compared to IgG4 wildtype Fc and IgG1 mAb controls (Table 3). This showed that specific mutant combinations could generate the desired structural fine-tuning to retain FcRn binding comparable to a wildtype IgG. We further investigated whether the FcRn binding affinity was augmented by the YTE mutations built into the library template. We found that the FcRn affinities of C4 and C4n differ by about 10-fold, similar to the difference in FcRn binding of IgG1 and IgG1 YTE. We also assessed the binding of C4 to other Fc interacting proteins. We observed the expected lack of binding to the FcγRI and FcγRIII receptors (data not shown), consistent with structural evidence that the lower hinge regions of both Fc chains are needed for FcγR interac- tion [35, 36]. However, to confirm the proper folding of the lower hinge region, we used a mutant form of the enzyme IdeS, known to bind to the lower hinge, and found that IdeS retained low micro-molar range binding affinity (3 μM) to C4, comparable to wildtype Fc. This supports the structural data that the lower hinge regions of our monomeric Fc and wildtype Fc are similar in conformation. Structure determination and refinement The peptide around the glycosylation site N297 spanning amino acids 290 through 300 and the glycans themselves appear somewhat less structured compared to the rest of the molecule (Fig 4B). The modeled biantennary carbohydrate moiety contains the core N-acetyl-D-glucosamine (GlcNAc or NAG) residue followed by another GlcNAc, followed by β-D-mannose (BMA) that splits the single carbohydrate chain into two chains. Each of these chains contains α-D-mannose (MAN) followed by GlcNAc, galactose (GAL), visible in one chain, and possibly a terminal sialic acid which is not visible in either of the chains. The core GlcNAc also has an α-L-Fucose residue attached. The CH3-CH3 interface of a wildtype IgG4 Fc is illustrated in Fig 4C. To disrupt the interac- tion between the two CH3 domains, we had selected 5 interface residues, L351, S354, T366, F405 and Y407, for our mutagenesis library. Although P395 is not at the interface, it was included in the library design to provide for possible backbone relaxation necessary to arrive at a stable monomeric Fc [17]. Our X-ray crystal structure for C4n confirmed a monomeric Fc devoid of CH3-CH3 dimerization. To understand the impact of the mutations in C4 or C4n that resulted in the monomer formation, the structure of the monomeric Fc was superimposed with each of the two chains of the wildtype IgG4 Fc (Fig 4D). It was revealed that the mutations introduced in C4 or C4n are indeed incompatible with Fc homodimer formation. The mutation T366R would have caused steric clashes with T366R and L351F of the opposing polypeptide chain. Similarly, F405R would have caused serious clash with the K392 side chain from the opposite Fc chain. The Y407E mutation removes the aromatic stacking interaction from the two tyrosine side chains in that position, favorable for dimerization. In addition, the glutamic acid side chain in position Y407E would have caused clashes with the same amino acid in the opposing chain. Monomeric Fc-fusion proteins To demonstrate that C4 can be used as a platform for monomeric Fc-fusion proteins, we gener- ated C4 fusions with the Fab and scFv fragments of the anti-cMet antibody Onartuzumab [6]. 10 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity Fig 4. Cartoon representation of the X-ray crystal structure of monomeric Fc C4n. (A) The protein chain and carbohydrates attached to N297 are shown in light salmon color. Light blue spheres indicate Zn atoms. (B) The model of the carbohydrate moiety attached to N297 superimposed onto the electron density map. Both of the possible terminal sialic acids and one of the galactose residues have no corresponding electron density map. The fucose attached to the core GlcNAc showed only partial electron density. (C) CH3-CH3 interface of wildtype IgG4 Fc showing side chains of the amino acids targeted in the mutagenesis library. Two interacting CH3 domains are shown in light blue and blue colors. (D) Superposition of two C4n structures (shown in light salmon and brown colors) onto wildtype IgG4 Fc to illustrate the effects of the mutations at the CH3-CH3 interface. Fig 4. Cartoon representation of the X-ray crystal structure of monomeric Fc C4n. (A) The protein chain and carbohydrates attached to N297 are shown in light salmon color. Light blue spheres indicate Zn atoms. (B) The model of the carbohydrate moiety attached to N297 superimposed onto the electron density map. Both of the possible terminal sialic acids and one of the galactose residues have no corresponding electron density map. The fucose attached to the core GlcNAc showed only partial electron density. (C) CH3-CH3 interface of wildtype IgG4 Fc showing side chains of the amino acids targeted in the mutagenesis library. Two interacting CH3 domains are shown in light blue and blue colors. (D) Superposition of two C4n structures (shown in light salmon and brown colors) onto wildtype IgG4 Fc to illustrate the effects of the mutations at the CH3-CH3 interface. Fig 4. Cartoon representation of the X-ray crystal structure of monomeric Fc C4n. (A) The protein chain and carbohydrates attached to N297 are shown in light salmon color. Light blue spheres indicate Zn atoms. (B) The model of the carbohydrate moiety attached to N297 superimposed onto the electron density map. Both of the possible terminal sialic acids and one of the galactose residues have no corresponding electron density map. Monomeric Fc-fusion proteins The fucose attached to the core GlcNAc showed only partial electron density. (C) CH3-CH3 interface of wildtype IgG4 Fc showing side chains of the amino acids targeted in the mutagenesis library. Two interacting CH3 domains are shown in light blue and blue colors. (D) Superposition of two C4n structures (shown in light salmon and brown colors) onto wildtype IgG4 Fc to illustrate the effects of the mutations at the CH3-CH3 interface. Fig 4. Cartoon representation of the X-ray crystal structure of monomeric Fc C4n. (A) The protein chain and carbohydrates attached to N297 are shown in light salmon color. Light blue spheres indicate Zn atoms. (B) The model of the carbohydrate moiety attached to N297 superimposed onto the electron density map. Both of the possible terminal sialic acids and one of the galactose residues have no corresponding electron density map. The fucose attached to the core GlcNAc showed only partial electron density. (C) CH3-CH3 interface of wildtype IgG4 Fc showing side chains of the amino acids targeted in the mutagenesis library. Two interacting CH3 domains are shown in light blue and blue colors. (D) Superposition of two C4n structures (shown in light salmon and brown colors) onto wildtype IgG4 Fc to illustrate the effects of the mutations at the CH3-CH3 interface. Fig 4. Cartoon representation of the X-ray crystal structure of monomeric Fc C4n. (A) The protein chain and carbohydrates attached to N297 are shown in light salmon color. Light blue spheres indicate Zn atoms. (B) The model of the carbohydrate moiety attached to N297 superimposed onto the electron density map. Both of the possible terminal sialic acids and one of the galactose residues have no corresponding electron density map. The fucose attached to the core GlcNAc showed only partial electron density. (C) CH3-CH3 interface of wildtype IgG4 Fc showing side chains of the amino acids targeted in the mutagenesis library. Two interacting CH3 domains are shown in light blue and blue colors. (D) Superposition of two C4n structures (shown in light salmon and brown colors) onto wildtype IgG4 Fc to illustrate the effects of the mutations at the CH3-CH3 interface. doi:10.1371/journal.pone.0160345.g004 11 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity Table 3. Equilibrium binding of Fc variants to human FcRn in avidity binding format. Monomeric Fc-fusion proteins Construct Apparent KD at pH 6 (nM) C4n 3560 C4 370 Onart-Fab-C4 160 Onart-scFv-C4 230 IgG4 Fc 140 IgG1 270 IgG1 YTE 39 doi:10.1371/journal.pone.0160345.t003 Table 3. Equilibrium binding of Fc variants to human FcRn in avidity binding format. cMet, a known cancer cell surface protein, is a member of the tyrosine kinase receptor family known to require monovalent targeting to avoid receptor agonism caused by receptor dimer- ization from bivalent antibodies. The C4-fusion proteins carrying either the Fab arm or the scFv arm of Onartuzumab retained binding to FcRn comparable to wildtype Fc (Table 3). Onart-Fab-C4 and Onart-scFv-C4 had high binding affinity to cMet, 0.5nM and 0.7nM, respectively. SEC-MALS demonstrated that these fusion constructs retained monomeric con- formation (Table 1). In cell growth inhibition assays with cMet-expressing Lovo cells, there was a marked agonist effect when cells were exposed to bivalent anti-cMet IgG, which is mostly absent in monovalent anti-cMet constructs (Fig 5A). This agonist effect due to the bivalent nature of anti-cMet illustrates a well-documented phenomenon in membrane protein targeting in literature and has limited the therapeutic uses of bivalent IgGs in certain receptor targeting contexts [37]. On the other hand, when the cells were stimulated with HGF both Onart-Fab- C4 and Onart-scFv-C4 molecules showed potent cMet-specific cell growth inhibition activity, similar to the monovalent heterodimeric Onartuzumab (Fig 5B). In vivo PK studies To demonstrate that FcRn binding affinity of C4 can lead to improved serum half-life of a monovalent fusion protein, we carried out in vivo pharmacokinetic studies of the Onart-Fab- C4 and Onart-scFv-C4 fusion proteins in human FcRn transgenic mice. The mice were dosed with 2.5 mg/kg of fusion proteins, and serum protein concentrations were determined by ELISA. Table 4 summarizes the relevant PK parameters. Compared to a Fab molecule alone, the monovalent Fc fusion proteins showed significant improvement in their PK properties (Fig 6), as manifested by 10-20-fold reduction in clearance (CL) and about 40-fold increase in ter- minal half-life (T1/2). This indicates that the C4-mediated human FcRn binding contributes to enhanced protein recycling. Compared to the wildtype IgG control, both monomeric Fc fusion proteins had a similar half-life of ~20 hours despite having 3-10-fold higher clearance. This was because the fusion proteins had a much larger volume of distribution (Vss), probably as a result of having smaller molecular weights compared to IgG. Discussion We have generated a monomeric Fc via a combination of rational design and in vitro evolution approaches. We used MALS and analytical ultracentrifugation analyses, in addition to size- exclusion chromatography, to characterize the monomeric Fc and used the crystal structure to demonstrate that our IgG4 based monomeric Fc exhibits an IgG Fc-like structure. The unique combination of these mutations, L351F, T366R, P395K, F405R and Y407E, was crucial in form- ing an Fc monomer with good monodispersity, even though other mutations of the same 12 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity Fig 5 Growth inhibition activities of anti-cMet-C4 fusion proteins and controls Lovo cells were tre Fig 5. Growth inhibition activities of anti-cMet-C4 fusion proteins and controls. Lovo cells were trea with Onart-Fab-C4 (dark green), Onart-scFv-C4 (light purple), one-armed heterodimeric antibody Onartuzumab (light green) [6], bivalent anti-cMet IgG (red) and negative control antibody (blue) in triplica and cell viability was measured by CellTiter-Glo assay after a 72-hour incubation. (A) Without ligand HGF treatment, there was a marked agonist effect when cells were exposed to bivalent anti-cMet IgG, an effec mostly absent in monovalent anti-cMet constructs. (B) With ligand treatment, anti-cMet targeting was sho to inhibit cell growth. Monomeric Fc fusions, Onart-Fab-C4 and Onart-scFc-C4 showed similar inhibitory effects as Onartuzumab. doi:10.1371/journal.pone.0160345.g005 Fig 5. Growth inhibition activities of anti-cMet-C4 fusion proteins and controls. Lovo cells were treated with Onart-Fab-C4 (dark green), Onart-scFv-C4 (light purple), one-armed heterodimeric antibody Onartuzumab (light green) [6], bivalent anti-cMet IgG (red) and negative control antibody (blue) in triplicates and cell viability was measured by CellTiter-Glo assay after a 72-hour incubation. (A) Without ligand HGF treatment, there was a marked agonist effect when cells were exposed to bivalent anti-cMet IgG, an effect mostly absent in monovalent anti-cMet constructs. (B) With ligand treatment, anti-cMet targeting was shown to inhibit cell growth. Monomeric Fc fusions, Onart-Fab-C4 and Onart-scFc-C4 showed similar inhibitory effects as Onartuzumab. d i 10 13 1/j l 016034 00 Fig 5. Growth inhibition activities of anti-cMet-C4 fusion proteins and controls. Lovo cells were treated i h O F b C4 (d k ) O F C4 (li h l ) d h di i ib d Fig 5. Growth inhibition activities of anti-cMet-C4 fusion proteins and controls. Our IgG4-YTE based monomeric Fc, C4, demonstrated human FcRn bind- ing affinity comparable to bivalent wildtype IgG4 or IgG1, which has not been previously PK parameters were determined by non-compartmental analysis using model 201. Cmax: peak concentration; AUC: the area under concentration; time curve: T1/2: terminal half-life; CL: clearance; Vss: volume in steady state. doi:10.1371/journal.pone.0160345.t004 Fig 6. Pharmacokinetic profiles of monomeric fusion proteins in hFcRn transgenic mice. Serum clearance curves are plotted for a wild-type IgG control (Motavizumab, red), Onart-Fab-C4 (blue), Onart- scFv-C4 (orange) and Fab control (purple) in hFcRn transgenic mice. Error bars, SE (standard error). C4 significantly extended the serum half-life of the fusion proteins. Compared to a wildtype IgG control, C4 fusions had an initial faster distribution clearance, possibly attributed to enhanced tissue distribution (Vss, Table 4) and then achieved similar serum half-life (T1/2, Table 4). doi:10.1371/journal.pone.0160345.g006 partmental analysis using model 201. Cmax: peak concentration; AUC: the area under concentration; time ss: volume in steady state. PK parameters were determined by non-compartmental analysis using model 201. Cmax: peak concentration; A curve: T1/2: terminal half-life; CL: clearance; Vss: volume in steady state. doi:10.1371/journal.pone.0160345.t004 protein can achieve equivalent serum half-life as an IgG. Taken together, our results suggest that the novel monomeric IgG4 Fc can be a promising therapeutic platform to extend the serum half-life of proteins in a monovalent format. There had been reported challenges in developing monomeric Fc, such as monomer insta- bility and reduced FcRn binding with disrupted CH3-CH3 dimer interface. For instance, there have been library-derived “monomeric” Fc mutants determined by SEC characterization that were later found to show only partial monomer formation when analyzed using light scattering [13, 14, 38]. Additionally, reduced FcRn binding has been reported by multiple studies attribut- ing to the lack of binding avidity to FcRn [11, 12]. We incorporated the YTE mutations (M252Y/S254T/T256E) in the CH2 domain to enhance FcRn binding [15, 39] into our mono- meric Fc template, and employed multiple biophysical and biochemical characterization meth- ods to screen for stable Fc monomers with IgG-like binding affinity to FcRn. This approach also sidestepped possible stability issues with adding YTE retrospectively to an engineered Fc monomer. The designed thermal selection steps during phage selection were found crucial in the enrichment for stable FcRn-binding Fc monomers as without them, sequence enrichment was not achievable. Discussion Lovo cells were treated with Onart-Fab-C4 (dark green), Onart-scFv-C4 (light purple), one-armed heterodimeric antibody Onartuzumab (light green) [6], bivalent anti-cMet IgG (red) and negative control antibody (blue) in triplicates and cell viability was measured by CellTiter-Glo assay after a 72-hour incubation. (A) Without ligand HGF treatment, there was a marked agonist effect when cells were exposed to bivalent anti-cMet IgG, an effect mostly absent in monovalent anti-cMet constructs. (B) With ligand treatment, anti-cMet targeting was shown to inhibit cell growth. Monomeric Fc fusions, Onart-Fab-C4 and Onart-scFc-C4 showed similar inhibitory effects as Onartuzumab. doi:10.1371/journal.pone.0160345.g005 doi:10.1371/journal.pone.0160345.g005 CH3-CH3 interface positions have been reported previously [12, 13, 17]. More importantly, using the IgG4-YTE based monomeric Fc, we have demonstrated that a monomeric Fc-fusion 13 / 18 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 IgG4 Monomeric Fc with Desirable Structure and Activity Table 4. In vivo mouse pharmacokinetic analysis of monomeric Fc-fusion proteins. Construct Dose Cmax AUCINF CL T1/2 Vss (mg/kg) (μg/mL) (hrμg/mL) (mL/hr/kg) (hr) (mL/kg) Fab 2.5 25 ± 5 8 ± 1 327 ± 58 0.6 ± 0.07 122 ± 28 Motavizumab 2.5 38 ± 6 877 ± 84 3 ± 0.3 18 ± 4 58 ± 9 Onart-Fab-C4 2.5 20 ± 2 160 ± 4 16 ± 0.4 21 ± 8 228 ± 49 Onart-scFv-C4 2.5 10 ± 7 92 ± 56 33 ± 15 24 ± 6 740 ± 410 PK parameters were determined by non-compartmental analysis using model 201. Cmax: peak concentration; AUC: the area under concentration; time curve: T1/2: terminal half-life; CL: clearance; Vss: volume in steady state. Table 4. In vivo mouse pharmacokinetic analysis of monomeric Fc-fusion proteins. PK parameters were determined by non-compartmental analysis using model 201. Cmax: peak concentration; AUC: the area under concentration; time curve: T1/2: terminal half-life; CL: clearance; Vss: volume in steady state. protein can achieve equivalent serum half-life as an IgG. Taken together, our results suggest that the novel monomeric IgG4 Fc can be a promising therapeutic platform to extend the serum half-life of proteins in a monovalent format. There had been reported challenges in developing monomeric Fc, such as monomer insta- bility and reduced FcRn binding with disrupted CH3-CH3 dimer interface. For instance, there have been library-derived “monomeric” Fc mutants determined by SEC characterization that were later found to show only partial monomer formation when analyzed using light scattering [13, 14, 38]. Additionally, reduced FcRn binding has been reported by multiple studies attribut- ing to the lack of binding avidity to FcRn [11, 12]. We incorporated the YTE mutations (M252Y/S254T/T256E) in the CH2 domain to enhance FcRn binding [15, 39] into our mono- meric Fc template, and employed multiple biophysical and biochemical characterization meth- ods to screen for stable Fc monomers with IgG-like binding affinity to FcRn. This approach also sidestepped possible stability issues with adding YTE retrospectively to an engineered Fc monomer. The designed thermal selection steps during phage selection were found crucial in the enrichment for stable FcRn-binding Fc monomers as without them, sequence enrichment was not achievable. Our IgG4-YTE based monomeric Fc, C4, demonstrated human FcRn bind- ing affinity comparable to bivalent wildtype IgG4 or IgG1, which has not been previously Fig 6. Pharmacokinetic profiles of monomeric fusion proteins in hFcRn transgenic mice. Serum clearance curves are plotted for a wild-type IgG control (Motavizumab, red), Onart-Fab-C4 (blue), Onart- scFv-C4 (orange) and Fab control (purple) in hFcRn transgenic mice. Error bars, SE (standard error). C4 significantly extended the serum half-life of the fusion proteins. Compared to a wildtype IgG control, C4 fusions had an initial faster distribution clearance, possibly attributed to enhanced tissue distribution (Vss, Table 4) and then achieved similar serum half-life (T1/2, Table 4). doi:10.1371/journal.pone.0160345.g006 PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 14 / 18 Fig 6. Pharmacokinetic profiles of monomeric fusion proteins in hFcRn transgenic mice. Serum clearance curves are plotted for a wild-type IgG control (Motavizumab, red), Onart-Fab-C4 (blue), Onart- scFv-C4 (orange) and Fab control (purple) in hFcRn transgenic mice. Error bars, SE (standard error). C4 significantly extended the serum half-life of the fusion proteins. Compared to a wildtype IgG control, C4 fusions had an initial faster distribution clearance, possibly attributed to enhanced tissue distribution (Vss, Table 4) and then achieved similar serum half-life (T1/2, Table 4). doi:10.1371/journal.pone.0160345.g006 Fig 6. Pharmacokinetic profiles of monomeric fusion proteins in hFcRn transgenic mice. Serum clearance curves are plotted for a wild-type IgG control (Motavizumab, red), Onart-Fab-C4 (blue), Onart- scFv-C4 (orange) and Fab control (purple) in hFcRn transgenic mice. Error bars, SE (standard error). C4 significantly extended the serum half-life of the fusion proteins. Compared to a wildtype IgG control, C4 fusions had an initial faster distribution clearance, possibly attributed to enhanced tissue distribution (Vss, Table 4) and then achieved similar serum half-life (T1/2, Table 4). doi:10.1371/journal.pone.0160345.g006 Fig 6. Pharmacokinetic profiles of monomeric fusion proteins in hFcRn transgenic mice. Serum clearance curves are plotted for a wild-type IgG control (Motavizumab, red), Onart-Fab-C4 (blue), Onart- scFv-C4 (orange) and Fab control (purple) in hFcRn transgenic mice. Error bars, SE (standard error). C4 significantly extended the serum half-life of the fusion proteins. Compared to a wildtype IgG control, C4 fusions had an initial faster distribution clearance, possibly attributed to enhanced tissue distribution (Vss, Table 4) and then achieved similar serum half-life (T1/2, Table 4). PLOS ONE \| DOI:10.1371/journal.pone.0160345 August 1, 2016 14 / 18 IgG4 Monomeric Fc with Desirable Structure and Activity demonstrated in a single monomeric Fc. Acknowledgments The authors would like to thank Xizhe Gao for assistance with generating pharmacokinetics calculations. We thank Gundo Diedrich, Qing Li and Jack Borrok for helpful discussions and provision of control in vivo data. We would also like to acknowledge Frank Comer and Vanessa Muniz-Medina for providing reagents and discussions on cMet growth inhibition assays. Supporting Information S1 Fig. The C4n structure model revealed a crystallization artefact related to Zn2+-medi- ated chain swapping. The conformation of the last 15 amino acids in the monomeric Fc struc- ture was changed substantially due to the presence of Zn2+ ions in the crystallization solution. As illustrated in (A), the 15 amino acids constituting the last β strand of Fc (in light salmon) were swapped with those from a symmetry related molecule (in green). (B) Detailed analysis showed that H433 and H435 have adopted a different position to coordinate a Zn2+ ion (shown in stereo). IgG4 Fc, in cyan, is superimposed with C4n structure in light salmon color, to show the extent of the conformational changes. (C) The tetrahedral coordination sphere of the Zn2+ ion is completed by H310 from the symmetry related molecule (in green) and a water molecule (W). (D) The electron density omit map of the chain swapping region is shown. (DOCX) Earlier studies had postulated that loss of avidity in Fc was the cause for reduced binding to FcRn [11, 12, 40]. Our data agreed with these observa- tions, as we showed that C4 demonstrated weaker binding to FcRn by about 10-fold compared to IgG1 YTE (Table 3). However, C4 affinity to FcRn is comparable to wildtype IgG, showing that it is possible to overcome the loss of avidity by the gain in affinity using YTE for mono- meric Fc. Our approach avoids the need for a tandem format to add avidity. In this work, monomeric Fc-fusion proteins with anti-cMet Fab and scFv were generated and showed excellent in vitro activity and biophysical properties. Furthermore, these C4-based fusion proteins demonstrated IgG-like serum half-life, suggesting the FcRn mediated recycling was fully effective. For targets such as cMet, where antibody-mediated receptor dimerization has undesirable effects, the technology has a broad application in providing a therapeutic platform and research tools. It offers a simplified single-chain strategy to achieve monomeric and monova- lent targeting, requiring only Protein A purification instead of additional purification processes necessary for monovalent heterodimeric Fc fusion proteins. PK analysis using human FcRn- transgenic mice demonstrated marked improvement of serum half-life over a Fab domain alone, which was cleared very rapidly because it is below the threshold of the renal glomeruli filtration size (~50–60 kDa) [41, 42]. In contrast, C4-fusion proteins, with only modest increase in size, resulted in over 10-fold slower clearance rate. More importantly, the serum half-life (T1/2) for Fab-C4 and scFv-C4 is similar to that of the IgG control. Furthermore, compared to IgG, the larger steady-state volume of distribution (Vss) suggests the monomeric Fc fusions may enhance tissue distribution. The role of reduced sizes of a half-life extending monomeric Fc fusion in tis- sue distribution can be further explored. Taken together, our novel monomeric IgG4 Fc construct and its fusion proteins demonstrated desirable in vitro activity and in vivo pharmacokinetics and could become a potential platform for generating monomeric Fc fusion proteins. References 1. 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https://openalex.org/W3002571200	OpenAlex	Open Science	CC-By	2,020	Family members and health care workers' perspectives on motivational factors of participation in physical activity for people with intellectual disability: A qualitative study	Henriette Michalsen	English	Spoken	8,364	12,058	Family members and health care workers’ perspectives on motivational factors of participation in physical activity for people with intellectual disability: A qualitative study H. Michalsen,1,2 S. C. Wangberg,3 A. Anke,1,2,4 G. Hartvigsen,5 L. Jaccheri6 & C. Arntzen1,7 1 Department of Rehabilitation, University Hospital of North Norway, Tromsø, Norway doi: 10.1111/jir.12716 doi: 10.1111/jir.12716 Journal of Intellectual Disability Research 259 VOLUME 64 PART 4 pp 259–270 APRIL 2020 1 Department of Rehabilitation, University Hospital of North Norway, Tromsø, Norway 2 Department of Clinical Medicine, Faculty of Health Sciences, University of Tromsø–The Artic University of Norway, Tromsø, Norway 3 Department of Health and Care Sciences, Faculty of Health Sciences, UiT – The Arctic University of Norway, Narvik, Norway 4 Institute of Health and Society, Research Centre for Habilitation and Rehabilitation Models and Services (CHARM), Faculty of Medicine, University of Oslo, Oslo, Norway 5 Department of Computer Science, Faculty of Science and Technology, UiT – The Artic University of Norway, Tromsø, Norway 6 Department of Computer Science, Faculty of Information Technology and Electrical Engineering, Norwegian University of Science and Technology (NTNU), Trondheim, Norway 7 Department of Health and Care Sciences, Centre for Care Research, Faculty of Health Sciences, North, UiT- The Arctic University of Norway, Tromsø, Norway 1 Department of Rehabilitation, University Hospital of North Norway, Tromsø, Norway 2 Department of Clinical Medicine, Faculty of Health Sciences, University of Tromsø–The Artic University of Norway, Norway Correspondence: Ms Henriette Michalsen, University Hospital of North Norway, Tromsø 9038, Norway (e-mail: henriette. michalsen@uit.no) tel.: +47 41500304. Abstract Results Three main themes were identiﬁed. According to support persons, motivation could be promoted at the individual level by fun, mastery, social setting, technology and knowledge about health behaviours. At a contextual level, physical activity was mediated by engagement with support individuals and available resources. At an interactional level, individuals were more motivated if the interaction was featured by joint activities, predictability and the use of rewards. Background People with intellectual disabilities (ID) have lower levels of physical activity than the general population. The aim of this study was to understand the motivational factors of participation in physical activity for people with ID from the perspectives of the family members and staff. Method An abductive qualitative design was used. Social Cognitive Theory constituted the theoretical frame of reference of the study. Two focus group interviews with health care workers and family members and two individual interviews with health care workers were conducted at their workplace. A thematic analysis was performed. Conclusions Motivation for participation in physical activity might be promoted at the individual, contextual and interactional levels. The interactions between individuals with ID and their support persons should work in a supportive way and strengthen mastery experiences. Support and engagement in the context could serve as a prerequisite for motivation and participation in physical activity and should be considered when Correspondence: Ms Henriette Michalsen, University Hospital of North Norway, Tromsø 9038, Norway (e-mail: henriette. michalsen@uit.no) tel.: +47 41500304. 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. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 260 H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual d developing interventions for physical activity for individuals with ID. Keywords intellectual disability, motivation, participation, physical activity, technology central Europe and the USA, and no studies have been conducted in the Arctic region. developing interventions for physical activity for individuals with ID. To understand the motivation and participation in PA for individuals with ID, studies have utilised health behavioural theories. Abstract In Social Cognitive Theory (SCT) (Bandura 1978), the concept of self- efﬁcacy states that the beliefs people have about their capabilities constitute a basis for action and also regulate motivation. These self-efﬁcacy beliefs can be strengthened through mastery experiences and social modelling inﬂuence. According to Self- Determination Theory (SDT) (Ryan and Deci 2000), motivation can be classiﬁed as (1) not present (amotivation), (2) leading to engagement in activity because it is enjoyable in itself (intrinsic) or (3) driven by external requests or rewards (extrinsic). Previous research has conﬁrmed that individuals with mild-to- borderline ID can display different levels of extrinsic motivation (Frielink et al. 2017). The focus of SDT on autonomy as intrinsically motivating could constitute an interesting perspective in this ﬁeld of study because of the frequent threats to personal control of people with ID (Bergström et al. 2014). Motivation can also be framed as a socioculturally constructed phenomenon (Daniels et al. 2007). In this frame of reference, motivation is always already embedded and shaped by particular contexts (Daniels et al. 2007). To understand the motivation and participation in PA for individuals with ID, studies have utilised health behavioural theories. In Social Cognitive Theory (SCT) (Bandura 1978), the concept of self- efﬁcacy states that the beliefs people have about their capabilities constitute a basis for action and also regulate motivation. These self-efﬁcacy beliefs can be strengthened through mastery experiences and social modelling inﬂuence. According to Self- Introduction Compared with the general population, individuals with intellectual disabilities (IDs) have worse health, lower levels of physical activity (PA) and higher barriers for participation in activities and access to health care (Walsh 2008; Evans et al. 2012; Hilgenkamp et al. 2012; Balogh et al. 2016; Temple et al. 2017; Wouters et al. 2019). PA is the term used to describe bodily movement produced by skeletal muscles that require energy expenditure (World Health Organization 2004, 2010) and can be further deﬁned by types of activities and intensity categories. As PA has positive effects on cardiovascular and psychosocial health factors, as well as brain health (Piercy et al. 2018), the identiﬁcation of effective interventions for use in everyday settings is important. Speciﬁcally, motivation of individuals with ID is a crucial concern (Hutzler and Korsensky 2010). Motivation refers to a driving force that elicits a particular action and affects the nature of an individual’s behaviour in both strength and persistence (Deckers 2005). Several studies have examined the barriers to and facilitators of participation in PA (Heller et al. 2003; Temple 2007; Mahy et al. 2010). Perceived self-efﬁcacy, social support and peer modelling are factors that facilitate the participation in PA of individuals with ID (Temple 2007; Bossink et al. 2017). The identiﬁed barriers to participation in PAs include lack of ﬁnancial, political and/or psychosocial support, as well as a lack of personal interest in PA and a lack of self-efﬁcacy (Dixon-Ibarra et al. 2017; Temple et al.2017). The perceptions of PA and other health behaviours among individuals with ID correspond to the carer and staff descriptions of barriers and preferences (Heller et al. 2003; Temple 2007; Mahy et al. 2010; Kuijken et al. 2016). Most research on PA for individuals with ID have focused on mild-to- moderate ID, and future research should include the study of individuals with more severe ID (Bossink et al. 2017). The majority of these studies are from As previous research indicates that interpersonal and other environmental factors contribute to the promotion of PA for individuals with ID, the authors aimed to explore how the individuals who interact with individuals with ID understand their participation in and motivation for PA. Our additional aims involved an investigation as to how PA could be facilitated in the Arctic regions and the inclusion of family members or staff who represent individuals with more severe ID. Family members We invited family members of individuals with varying degrees of intellectual disability, age and family relationship, as well as living conditions, with the help of user organisations and the networks of the authors. Table 2 provides a full overview of the participants. For the purpose of anonymisation, some characteristics of the participants have been changed. Participants A purposeful sampling strategy was used to recruit participants (Patton 2015). Health care workers and family members were interviewed separately, as we believed that this would create a more open environment for the individuals to speak freely about the motivational factors and barriers to participation in PA (Malterud 2012). The study was approved by the Committee for Medical Research Ethics (No. 2016/1770). Design An abductive qualitative design was utilised (Kardorff et al. 2004). To understand new aspects of participation and motivation for PA, it was relevant to use an explorative, empirically driven approach, with portions of the existing literature on the topic as a basis. Health care workers Health care workers and services, we supplemented the data with two individual in-depth interviews with health care workers (Kvale 2015). The interview guide and analyses of results were inspired from the SCT (Bandura 1978), deﬁnition of motivation from the SDT (Ryan and Deci 2000), the sociocultural learning theory (Daniels et al. 2007) and previous research on barriers and facilitators for PA (Temple 2007; Mahy et al. 2010). The interview guide helped structure the interview and ensured that key areas were covered and discussed. Central themes were to describe a regular day and challenges, preferences, facilitators and motivating factors for engaging in physical activity. We invited participants with different roles (leaders/ nonleaders) who worked in four different settings (specialised and community-based intellectual disability services, as well as day care centres). The services are predominantly for people with moderate, severe or profound ID. A focus group interview with three participants was conducted (Morgan 1997) in which participants provided important information and were regarded as representing information-rich cases (Patton 2015). Additionally, we conducted two in-depth individual interviews (Kvale 2015) with the leader of a community residence and a representative of a day care centre. The interviews took place at their work place, and the main topics from the focus group interview guide were used. Table 1 provides an overview of the participants. An abductive qualitative design was utilised (Kardorff et al. 2004). To understand new aspects of participation and motivation for PA, it was relevant to use an explorative, empirically driven approach, with portions of the existing literature on the topic as a basis. Methods We conducted two semistructured focus group interviews with family members and health care workers. Focus group interviews are suitable because they facilitate dialogue and the elaboration of experiences and ideas among colleagues (Malterud 2012) and is useful for exploring speciﬁc sets of issues as well as permitting the researcher to use the group interactions as research data (Kitzinger 1994). To ensure greater variety in the sample of professionals © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 261 Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disab H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability Data collection Throughout the analysis, the SCT concept and sociocultural perspectives became central to understanding motivation as a relational phenomenon and the role of the context in initiating and adapting for individual participation in PA. The authors were from different professions with competence in rehabilitation medicine, behavioural change theories, use of technology and qualitative research methodology, and the setting was an exploration of motivational factors for possible later use in technology-supported activity. Regarding the analysis, the authors had no previous experience with the research problem at hand and few preconceptions and expectations prior to data collection and analysis. transformed into speciﬁc codes, which were compared by differences and similarities and condensed into meaningful categories and subcategories. Through discussions among the authors and other researchers and with feedback from the reference group, the main themes were identiﬁed by grouping similar subthemes and linking them to theories and perspectives. Throughout the analysis, the SCT concept and sociocultural perspectives became central to understanding motivation as a relational phenomenon and the role of the context in initiating and adapting for individual participation in PA. The authors were from different professions with competence in rehabilitation medicine, behavioural change theories, use of technology and qualitative research methodology, and the setting was an exploration of motivational factors for possible later use in technology-supported activity. Regarding the analysis, the authors had no previous experience with the research problem at hand and few preconceptions and expectations prior to data collection and analysis. All participants were enthusiastic about the topic, which inﬂuenced the dynamic of the discussions and the richness of the data. As the introductory questions stimulated dialogue and discussion, the interview guide was mainly used as a checklist to ensure that key topics were covered. All participants were found to be information-rich cases. The interviews were recorded using an MP3 recording device and transcribed verbatim by H. M. After the individual interviews, the transcripts were examined, and the investigators concluded that data saturation was reached. Data analysis After the interviews, preliminary analyses were presented to a reference group (researchers in the project group and some of the family members and health care workers from interviews) for their feedback before continuing with the analysis. Preliminary results were also presented at a seminar with experienced researchers, who commented on and further improved the analysis before it was ﬁnalised, to ensure the dependability and trustworthiness of the results (Tjora 2012). A thematic analysis (Greenhalgh and Taylor 1997) was then conducted. The interview transcripts were read several times by the ﬁrst author (H. M.) to identify emerging themes. The interviews were then read and commented by A. A. On the one hand, C. A. read and commented on the ﬁrst analysis of the material. The text of the transcriptions was After the interviews, preliminary analyses were presented to a reference group (researchers in the project group and some of the family members and health care workers from interviews) for their feedback before continuing with the analysis. Preliminary results were also presented at a seminar with experienced researchers, who commented on and further improved the analysis before it was ﬁnalised, to ensure the dependability and trustworthiness of the results (Tjora 2012). Data collection The interviews lasted between 1 and 2 h. C. A. led the focus group interviews (as the moderator) and H. M. observed the interviews, took notes and clariﬁed questions throughout the interviews (as the secretary) (Malterud 2012). The individual interviews were led by H. M. Table 1 Sociodemographic characteristics of health care workers Profession Gender Work experience (years) Workplace Data collection method Intellectual disability nurse Male 8, 5 Home-based care Focus group Intellectual disability nurse Male 9 Home-based care Focus group Physiotherapist Female 2, 5 Specialised health care Focus group Leader of community residence Female 7 Community residence† Individual interview Teacher Female 3, 5 Day care centre Individual interview †Community residence: municipal service that provides supportive care around-the-clock. Table 1 Sociodemographic characteristics of health care workers © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 262 Table 2 Family members’ relations to the individuals with intellectual disabilities and characteristics of the individuals with intellectual disabilities. Some characteristics are changed for the purpose of anonymisation Relation to person with ID Gender of person with ID Age of person with ID Living conditions of person with ID Level of ID Mother Male 20 At home with family Severe ID Mother Male 19 At home with family Moderate ID Father Female 25 Supported living† Moderate ID Father Male 16 At home with family Profound ID Father Male 22 Community residence‡ Moderate ID Mother Male 13 At home with family Moderate ID Sister Male 52 Community residence‡ Severe ID Father Female 23 Supported living Severe ID ID, intellectual disability. †Supported living: individuals living in their own apartment with part-time supervision from staff. ‡Community residence: municipal service that provides supportive care around-the-clock ID, intellectual disability. †Supported living: individuals living in their own apartment with part-time supervision from staff. ‡Community residence: municipal service that provides supportive care around-the-clock transformed into speciﬁc codes, which were compared by differences and similarities and condensed into meaningful categories and subcategories. Through discussions among the authors and other researchers and with feedback from the reference group, the main themes were identiﬁed by grouping similar subthemes and linking them to theories and perspectives. Results (leader of community residence) Main themes Subthemes Codes Individual level Motivation seldom the main challenge Difﬁculty initiating activities Motivation often present Varying levels of exhaustion Fun, social rewards and technology Being social in activities is rewarding for many Purpose of the activity is important Use of technology rewarding Knowledge Lack of knowledge in what constitutes good health Reduced knowledge about healthy behaviours in support persons Health education for individuals with ID not a priority for support persons Contextual level Support person engagement Many factors depend on support person’s interest in physical activity Engagement in support person increases levels of physical activity Available resources and weather considerations Not enough resources/personnel available Variation in weather conditions in the arctic climate is a barrier Presentation of activity Willingness to join activities depends on a positive presentation of activities Interaction between levels Being together in activities Giving support in activities by participating together with individuals with ID is important Creating predictability It is important to create predictability in the physical activity Use of rewards Rewards can be used to reinforce the physical activities Showing achievements to others is very rewarding © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Results We identiﬁed one overarching theme (motivation for participation), three main categories and nine subthemes. The main ﬁnding was that motivation could be socially constructed through mediation of engagement in activities and external support. An overview of the ﬁndings is given in Table 3 and summarised in Fig. 1. A thematic analysis (Greenhalgh and Taylor 1997) was then conducted. The interview transcripts were read several times by the ﬁrst author (H. M.) to identify emerging themes. The interviews were then read and commented by A. A. On the one hand, C. A. read and commented on the ﬁrst analysis of the material. The text of the transcriptions was © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 263 Table 3 Summary of results Individual level Motivation is not always the challenge. Both family members and health care workers identiﬁed the main issue as a lack of initiation of PA for individuals with ID, rather than a lack of motivation. Understanding how people with ID were motivated to participate in PA was challenging and could vary in different situations, but most of the participants believed that motivating the individuals to participate was not the most difﬁcult part. … there are many, if I speak on behalf of the institution I work in, who want to go hiking and be out in the nature, and who have the motivation to participate. Individual level Motivation is not always the challenge. Both family members and health care workers identiﬁed the main issue as a lack of initiation of PA for individuals with ID, rather than a lack of motivation. Understanding how people with ID were motivated to participate in PA was challenging and could vary in different situations, but most of … there are many, if I speak on behalf of the institution I work in, who want to go hiking and be out in the nature, and who have the motivation to participate. (leader of community residence) © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 264 Figure 1. Results from the qualitative study. Themes and structure. Figure 1. Results from the qualitative study. Themes and structure. context, described as being part of a team, meeting peers and identifying with a group. I am of the impression that Sophie could walk on water if she was motivated to do so. It is just very difﬁcult for me to ﬁgure out what actually motivates her nowadays … (mother) I am of the impression that Sophie could walk on water if she was motivated to do so. It is just very difﬁcult for me to ﬁgure out what actually motivates her nowadays … (mother) He loves his friends and being part of a team. He could go to the North Pole if that was the case, as long as he was part of the team … (mother) Difﬁculties in completing an activity could reduce motivation, such as problems with getting dressed, difﬁculties in travelling to the site of the activity, or problems in executing the physical movements. Difﬁculties in completing an activity could reduce motivation, such as problems with getting dressed, difﬁculties in travelling to the site of the activity, or problems in executing the physical movements. Several individuals were more motivated to perform PA when the activity had a different purpose than the activity itself, such as enjoying a meal at the end of a trip or doing practical chores as part of the PA. It depends on practical stuff, like how to put the skis on. That is the most difﬁcult thing when going skiing. Individual level For most of the ski trips that are unsuccessful, everything goes wrong when he struggles with putting his ski boots or skis on, and gets tired and gives up. (sister) … we always have to ﬁnd things to recycle and walk by a container and throw it in. And there is no lunch before we have been out walking for a while, and then it’s just a matter of how long you can stretch it. (mother) The participants also questioned the level of intensity of PA, with many stating that individuals with ID participated in activities but rarely achieved a level of moderate or vigorous PA. … it’s the same for our boy, as I hear others talk about, that it should be another purpose with the activity than the activity itself. The best is to bring a good lunch to eat on the top of a mountain or do practical chores like walking by the grocery store to a recycling bin. (father) … it’s the same for our boy, as I hear others talk about, that it should be another purpose with the activity than the activity itself. The best is to bring a good lunch to eat on the top of a mountain or do practical chores like walking by the grocery store to a recycling bin. (father) … it’s difﬁcult ﬁnding physical activities that would improve her ﬁtness shape. She rarely breaks a sweat or feel out of breath when she exercises … (mother) Several individuals enjoyed swimming and horseback riding. Others enjoyed listening to music while moving around or while watching a programme on a tablet or a TV. It was reported that most of the individuals with ID (especially the © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Knowledge, interests and misunderstandings Some of the health care workers mentioned that as individuals with ID may lack knowledge of risk factors for ill health, they could beneﬁt from learning more about health and how PA will have a positive impact on their lives. In some cases, individuals with ID would misinterpret bodily signals from PA as signs of sickness and associate the activity with something negative and harmful. … When it comes to those with ID, maybe moderate or severe ID, I think the staff plays a much larger role. Because the staff needs to be motivated as well. (physiotherapist). Most of the family members were worried that the staff would never show the same interest and enthusiasm as the family members did in supporting a physically active lifestyle. … they might be frightened if they feel their heart pounding harder than they are used to, and think that they are ill or something … I think it has to do with the knowledge basis they have. How much knowledge they have will affect how they interpret the body’s signals. If they have more knowledge about how the body reacts to excessive physical activity, they might not react this way. (leader of community residence) … it is hard to make the staff see his skills when it comes to physical things. It’s just too easy to see his epilepsy as a problem and be anxious about it. (father) … it is hard to make the staff see his skills when it comes to physical things. It’s just too easy to see his epilepsy as a problem and be anxious about it. (father) Several individuals also mentioned that they wished that they had an ‘instruction manual’ for their family member, so that staff would know how to motivate and make adjustments so that the individual with ID could be more physically active. In contrast, family members were more concerned about the level of knowledge, interest and enthusiasm that health care workers had concerning PA and how that could inﬂuence the individuals with ID. They believed that it would not necessarily help individuals with ID gain knowledge. … I have said to the staff ‘come join us on cross country skiing and see how easily it’s done’. And some actually do so. Fun, social rewards and technology Individuals with ID were more likely to participate in PA when the activity was fun and enjoyable. Activities were observed to be enjoyable if they were in a social © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 265 Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disab H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability younger individuals) were interested in technology. Some used technological devices in PA settings. motivation and execution of PA. The attitudes concerning PA and health in the people close to the individuals with ID were observed to be important predictors of PA by all participants. She has two, an iPhone she uses to ﬁnd things she wants to watch on her iPad (music videos), and a PC as well … (mother).… he is allowed to sit on a small trampoline whilst watching his iPad or the TV, and if he gets excited, he starts jumping. And he can watch the ﬁlm to the end whilst jumping if the ﬁlm is very funny… (father) … Some of my staff are really active and like to do new things, and if they do not manage to get the user interested, they try a different approach. They always bring the right clothes according to the weather … I think it has a lot to do with the attitude of the staff. Do not get me wrong, but I think if the staff want to be inside and relax, it is very easy to do so. (leader of community residence) Available resources and weather considerations Available resources and weather considerations In the group of health care workers, the availability of resources was often mentioned as an important condition to be able to take individuals with ID to perform PA. Insufﬁcient stafﬁng, a lack of time and Contextual level In the group of health care workers, the availability of resources was often mentioned as an important condition to be able to take individuals with ID to perform PA. Insufﬁcient stafﬁng, a lack of time and Knowledge, interests and misunderstandings But then you have people working on shift and some are quite old and … it’s just very complicated and exhausting for the family to always teach new staff how to do things. (sister). … he cannot just understand for himself why it is important to exercise, that it is important to be physically active. He can say that he is using energy when he is active, but that is because I have triggered him to think that it is important. (mother) Interaction between individual and context Examples of cites in relation to subthemes are given in Table 4. Being together in activities Being together in activities Many of the participants, especially the health care workers, pointed out that if they identiﬁed the right way of presenting the activity to the individual, then it was more likely to motivate the individual to participate in that activity. By simply asking the Many of the stories concerning success factors for PA focused on the interaction between individuals with ID and their environments. If the support personnel and/or family members were successful in inﬂuencing Table 4 Examples of interview cites from the theme Interaction between individual and context Theme: Interaction between individual and context Theme: Interaction between individual and context Subthemes Cites Being together in activities … the support person should not just get the individual into the pool – he should join him in the pool. In our home, the support person shows up, brings my son to the arena and sits on the scaffold and pays attention to what my son does. I wish that he could actually join him in his workout. Many of the individuals with ID will have much greater advantages and better workouts if they had their support person with them all the time to motivate them and help them. (father) Creating predictability … one thing I’ve been thinking a lot about, and that is making the activity predictable is very important. And there is of course a challenge to understand how this would work. After a while, in my experience, he has learned that if he is prepared for what’s coming next, and we help him to do so, it’s so much more fun for him to participate. (mother) Receiving rewards … in my experience it seems like it should be something there. If it’s not something very concrete. My son needs to understand why he should do different things. What the goal or intention is for doing these things. Either it is the good lunch afterwards or he can use his iPad when we are done doing the activity. (mother) © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Cites … the support person should not just get the individual into the pool – he should join him in the pool. In our home, the support person shows up, brings my son to the arena and sits on the scaffold and pays attention to what my son does. I wish that he could actually join him in his workout. Many of the individuals with ID will have much greater advantages and better workouts if they had their support person with them all the time to motivate them and help them. (father) … one thing I’ve been thinking a lot about, and that is making the activity predictable is very important. And there is of course a challenge to understand how this would work. Support person engagement Support person engagement All participants agreed that the contexts surrounding the individuals with ID played a large role in their © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 266 lsen et al. • Motivational factors of participation in physical activity for people with intellectual disability insufﬁcient preparation for changing weather conditions were mentioned as barriers. individuals with ID if they wanted to exercise, the workers observed that this would most often lead to a negative response. The challenge is for those who do not have many resources around them. You’re at work and have maybe ﬁve people to look after during your shift. And everyone should get dinner and have individually adjusted activities in the afternoon. So that is where the problem is, because one of the users might want to go swimming and you cannot leave the other four users behind. And is just hard sometimes … (leader of community residence) … asking ‘do you want to go for a walk’, you are more likely to get a negative reply, because it is a personal question. If you say ‘let us go for a walk’ or ‘what should we do today? Should we go for a walk?’, it helps motivate the individual to participate. It’s easier to bring them with us. (teacher) © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Presentation of activity Being together in activities Creating predictability Making the PA predictable and creating a sense of control for individuals with ID were also important aspects of motivation for PA. The use of different tools, such as visual support in communicating about the PA, going to the arena and examining it beforehand or making PA a part of their daily routine, also helped the individual to feel more in control. One father said that it was more likely that his son or his peers would feel excited about participating in athletics when they had been there a few times before and knew what they were getting into. Consistent with previous studies (Temple 2007; Mahy et al. 2010), motivation is dependent on how fun and interesting the PA is perceived to be. It is likely to believe that the motivation individuals with ID show towards PA is displayed as an interest for the activity or enjoyment during participation. Another potentially useful ﬁnding for intervention development is the interest in technology by the group of youths and young adults with ID. Technological developments, like serious games, have previously been proven to be effective interventions for improving social, practical and conceptual skills for individuals with ID (Tsikinas and Xinogalos 2018). Receiving rewards Rewards could make PA more interesting. However, many of the participants believed that it was difﬁcult to identify rewards that were not unhealthy foods. The other rewards that were mentioned as effective included a high degree of praise or positive feedback to the individuals. If they received a medal after participating in PA, the reward was not the medal itself, but the praise when showing the medal to the staff or family members afterwards. The results demonstrate that engagement in PA by support persons is particularly important. Support persons or others who interact with individuals with ID on a daily basis should have an initial interest for PA or be concerned about increasing PA levels for the individual with ID. Heller et al. (2003) underlines that support persons should be motivated to perform PA themselves, as this would inﬂuence the individual with ID to be more active. Activities should be planned in cooperation between support persons and the individual with ID while considering motivational factors. In contrast, a lack of available resources functions as a contextual barrier for participation (Mahy et al. 2010; Bergström 2014; Bergström et al. Theme: Interaction between individual and context the individual with ID in a motivating way, the individual would have a positive attitude towards the activity. Motivational support could include performing the PA with the individual, providing positive feedback, cheering the individuals on when they were active and reinforcing their positive experiences with the PA. Other types of support, such as practical support and economical support, were also mentioned as contributing to participation in PA. Theme: Interaction between individual and context After a while, in my experience, he has learned that if he is prepared for what’s coming next, and we help him to do so, it’s so much more fun for him to participate. (mother) … in my experience it seems like it should be something there. If it’s not something very concrete. My son needs to understand why he should do different things. What the goal or intention is for doing these things. Either it is the good lunch afterwards or he can use his iPad when we are done doing the activity. (mother) © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 267 H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability In this study, the main source of information was from the environment around the individual with ID, not the individual himself or herself. By emphasising the sociocultural frame of reference (Daniels et al. 2007), we found that motivation is an interpersonal emerging phenomenon continuously shaped by particular practice. This makes it is difﬁcult to capture the intrinsic motivational factors involved in participating in physical activity. Positive emotions such as engagement and excitement might be observable features that can be interpreted as an expression of motivation. However, based on our results, the deﬁnition of motivation according to the SDT (Ryan and Deci 2000) does not capture the complexity of this cognitive feature. Motivation can be seen as a socially driven interaction between individuals with ID and their support persons that can increase or decrease depending on the responses from the individual or the context. These ﬁndings can be important when developing interventions for PA and adjusting the everyday contexts for individuals with ID. the individual with ID in a motivating way, the individual would have a positive attitude towards the activity. Motivational support could include performing the PA with the individual, providing positive feedback, cheering the individuals on when they were active and reinforcing their positive experiences with the PA. Other types of support, such as practical support and economical support, were also mentioned as contributing to participation in PA. Discussion The ﬁndings of the current study suggest that individuals with ID could be more motivated to participate in PA if the activities occur as an interaction between the individual and their context, which could be mediated by engagement with support people and available resources. At an individual level, individuals with ID can be motivated for PA if they experience the activity as fun, occurring in a social setting or used in combination with technology. An interaction through joint participation in activities, predictability and the use of rewards appears to have the strongest inﬂuence on participation in PA. © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 268 Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disab H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual disability 2; Kuijken et al. 2016). Inadequate stafﬁng, little knowledge about the health beneﬁts of PA and unclear PA policies in the community residence or day care centre could inﬂuence the participation in PA of individuals with ID. Dixon-Ibarra et al. (2017) argue that a lack of clear PA policies in group home settings creates a barrier to health-promoting environments for people with ID. Individuals with ID rely heavily on support from others in their everyday life, and possibilities to maintain good health will therefore vary depending on the services and care that they receive (Krahn et al. 2006). participation through the perspectives of family members and health care workers, we did not interview individuals with ID themselves. Future studies could explore the motivation for PA from the perspective of less active participants and could also focus on interviews with individuals with ID themselves. Nevertheless, our ﬁndings are consistent with previous studies (Heller et al. 2003; Mahy et al. 2010). Furthermore, while there were relatively few participants who completely contributed with information, a strength of the current study is that the participants represented a varied sample of individuals with ID. These ﬁndings could be relevant outside this research context. The ﬁndings are consistent with SCT (Bandura et al. Study strength and limitations The present study had several limitations. Most of the participants who participated in the study had an initial interest in PA, as well as being physically active themselves. Additionally, as the initial aim of this study was to understand the motivation for PA Acknowledgements The authors would like to thank all participants who were involved in the study for contributing to such an important research topic. Conclusion Promotion of PA should include both individuals with ID, family members and staff, and it should work in a supportive way to strengthen mastery experiences. Fun, social rewards, predictability and the use of technology as motivational factors for participation in PA on an individual level are interesting possibilities that should be explored further. The promotion of PA in individuals with ID should also take a contextual approach by exploiting the cooperation and interaction between individuals with ID and their support persons. Future PA interventions for this group should focus on engaging the support persons in physically activities themselves and on increasing their knowledge about the importance of PA and ways to motivate individuals with ID. Arctic weather conditions may require extra human resources and equipment for outdoor activities. Many participants found the planning and execution of outdoor PA during winter to be challenging. For both the support person and the individual with ID, changing weather conditions and preparations for outdoors could disturb motivation and participation in PA. Individuals with ID who displayed sedentary behaviour often mentioned weather as a barrier. This research suggests that individuals with ID may not have the strategies to cope with this adversity alone and need assistance to overcome this barrier. Discussion 1999), in that self-efﬁcacy appeared to be related to the intention to perform PA. Self-efﬁcacy was positively inﬂuenced by previous mastery experience with the PA, as well as modelling by other individuals. This emphasises the need for preparation and explanation of the activity to ensure high self-efﬁcacy and opportunities to strengthen it further through more mastery experiences. Conversely, the results of this study demonstrated that where the individuals with ID had a negative experience with PA (e.g. skiing) early in the process, then many individuals lost their motivation and stopped participating in the activity. Therefore, the planning for progressive mastery by the individual is essential. Conﬂict of Interest The authors declare that they have no conﬂict of interest. References Hutzler Y. & Korsensky O. 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(1997) How to read a paper: papers that go beyond numbers (qualitative research). BMJ [British Medical Journal] 315, 740–3. Tjora A. (2012) Kvalitative forskningsmetoder i praksis. 2. utgave. Gyldendal norsk forlag AS, Oslo. Heller T., Hsieh K. & Rimmer J. (2003) Barriers and supports for exercise participation among adults with Down Syndrome. Journal of Gerontological Social Work 38, 161–78. Tsikinas, S., & Xinogalos, S. (2018, 17–20 April 2018). Designing effective serious games for people with intellectual disabilities. Paper presented at the 2018 IEEE Global Engineering Education Conference (EDUCON), Tenerife, Spain. Hilgenkamp T. I., van Wijck R. & Evenhuis H. M. (2012) Low physical ﬁtness levels in older adults with ID: results of the HA-ID study. Research in Developmental Disabilities 33, 1048–58. Walsh P. N. (2008) Health indicators and intellectual disability. Current Opinion in Psychiatry 21, 474–8. © 2020 The Authors. Wouters M., Evenhuis H. M. & Hilgenkamp T. I. M. (2019) Physical activity levels of children and adolescents with moderate-to-severe intellectual disability. Journal of Applied Research in Intellectual Disabilities 32, 131–42. World Health Organization (2010) Global Recommendations on Physical Activity for Health. World Health Organization, Geneva, Switzerland. World Health Organization (2004) Global Strategy on Diet, Physical Activity and Health. World Health Organization. © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd References Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Scientiﬁc Study of Intellectual and Developmental Disibilities and John Wiley & Sons Ltd © 2020 The Authors. Journal of Intellectual Disability Research published by MENCAP and International Association of the Journal of Intellectual Disability Research VOLUME 64 PART 4 APRIL 2020 270 H. Michalsen et al. • Motivational factors of participation in physical activity for people with intellectual d World Health Organization (2004) Global Strategy on Diet, Physical Activity and Health. World Health Organization. World Health Organization (2010) Global Recommendations on Physical Activity for Health. World Health Organization, Geneva, Switzerland. Wouters M., Evenhuis H. M. & Hilgenkamp T. I. M. (2019) Physical activity levels of children and adolescents with moderate-to-severe intellectual disability. Journal of Applied Research in Intellectual Disabilities 32, 131–42. Accepted 13 January 2020 Accepted 13 January 2020	2,478
https://openalex.org/W1928706541	OpenAlex	Open Science	CC-By	2,002	Integrated multimedia processing for topic segmentation and classification	Radu Jasinschi	English	Spoken	4,042	7,153	ABSTRACT In this paper we describe integrated multimedia processing for Video Scout, a system that segments and indexes TV programs according to their audio, visual, and transcript information. Video Scout represents a future direction for personal video recorders. In addition to using electronic program guide metadata and a user proﬁle, Scout allows the users to request speciﬁc topics within a program. For example, users can request the video clip of the Pres- ident speaking from a half-hour news program. In this paper we describe integrated multimedia processing for Video Scout, a system that segments and indexes TV programs according to their audio, visual, and transcript information. Video Scout represents a future direction for personal video recorders. Content based video analysis and processing has been an active topic of research [5]. However, there are very few initial approaches to multimodal processing of audio, visual, and transcript informa- tion. The majority of content-based systems include Query-By- Image-and-Video-Content (QBIC), VisualGrep, DVL of AT&T, InforMedia, VideoQ, MoCA, Vibe, and CONIVAS. In particular, the InforMedia, MoCA, and VideoQ systems are more related to Video Scout. In addition to using electronic program guide metadata and a user proﬁle, Scout allows the users to request speciﬁc topics within a program. For example, users can request the video clip of the Pres- ident speaking from a half-hour news program. Video Scout has three modules: (i) Video Pre-Processing, (ii) Segmentation and Indexing, and (iii) Storage and User Interface. Segmentation and Indexing, the core of the system, incorporates a Bayesian framework that integrates information from the audio, visual, and transcript (closed captions) domains. This framework uses three layers to process low, mid, and high-level multimedia information. The high-level layer generates semantic information about TV program topics. This paper describes the elements of the system and presents results from running Video Scout on real TV programs. However, there are very few initial approaches to multimodal processing of audio visual and transcript information in video anal- ysis. Vasconcelos and Lippman [6] propose a statistical model that uses video content structure information to perform video shot detection and semantic characterization. Rui et al. [9] present an approach that uses low level features in audio to detect ex- cited speech and a baseball hit within a probabilistic framework for automatic highlight extraction. Syeda-Mahmood et al. [8] present event detection in multimedia presentations from teaching and training videos. 1. INTRODUCTION Personal Video Recorders (PVRs), such as TiVo [3], use electronic program guide metadata to automatically select and store whole TV programs based on users’ proﬁles. Video Scout represents a future direction for PVRs. Scout analyzes the audio, visual, and transcript (closed captions) content in order to segment and index TV programs. In addition, Scout allows users to request topic level information. For example: users can request speeches by the Pres- ident, and when the nightly news is broadcast, Scout can extract the President’s speech from the half-hour news broadcast. This paper describes the elements of the system and presents results from running Video Scout on real TV programs. The main advantage of our system over the previously men- tioned approaches is the use of multilevel audio, visual, and tran- script processing for story segmentation and topic classiﬁcation as opposed to indexing video based on low-level features. In addition, our system uses multimodal processing for consumer applications, which have been ignored by previously described systems. ABSTRACT They use a probabilistic model that exploits the co-occurrence of visual and audio events. Another approach based on the observation that semantic concepts in videos interact and appear in context is proposed by Naphade [1]. To model this contextual interaction explicitly, a probabilistic graphical network of multijects or a multinet is proposed. R. S. Jasinschi, N. Dimitrova, T. McGee, L. Agnihotri, J. Zimmerman, and D. Li Philips Research, 345 Scarborough Road, Briarcliff Manor, NY 10510 3. SYSTEM OVERVIEW Video Scout employs three interconnected modules: (i) Video Pre-Processing; (ii) Segmentation and Indexing; and (iii) Stor- age and User Interface. Segmentation and Indexing, the core of the system, uses a three layered probabilistic system, called the Bayesian Engine (BE). The BE integrates information from the audio, visual, and transcript domains in order to generate seman- tically indexed information from TV program segments. We im- plemented Video Scout as an end-to-end system, and in its current state it segments, indexes, and stores TV programs for ﬁnancial news and for talk shows. Figure 1 illustrates Video Scout’s three modules. Video Pre-Processing, receives raw video that has been, de-muxed and decoded as nec- essary. This generates three streams: audio, visual, and transcript. Decoding is optional and is only implemented when further video processing is done in the uncompressed domain. Segmentation and Indexing, takes these three streams as input. It also receives electronic program guide metadata that describes the TV programs. This metadata contains general information such as channel, time, title, cast, etc. In addition, Segmentation and Indexing has input from program proﬁles (PP) and the content program proﬁles (CPP). Section 2 offers a brief overview of related work. Section 3 gives a system overview of Scout. Section 4 describes the Video Pre-Processing module. Section 5 presents the Segmentation and Indexing module. Section 6 describes topic segmentation and clas- siﬁcation. Section 7 shares our experimental results. And in Sec- tion 8 we draw conclusions from this work. Users specify their program preferences via the PP, which can be used to generate recommendations about TV programs. The CPP allows users to input topic level preferences. 5. SEGMENTATION AND INDEXING Indexing Segmentation Transcript C P E Audio Visual Figure 2 contains an illustration of the BE, the core of Segmenta- tion and Indexing. We organized this probabilistic framework in three consecutive layers: low, mid and high-level. Each layer has nodes with associated probabilities. Arrows between the nodes in- dicate a causal relationship. audio Audio Visual High-Level Mid-Level Low-Level Transcript Video In Color Edge Shape Parameters Video Text Silence Noise Speech Music Sp+No Sp+Sp Sp+Mu Keyword Celebrities Financial Commercial CC Keyframe Face CC Categories Fig. 2. The three layers of the Segmentation and Indexing module of Video Scout. Storage User Input Fig. 1. The three modules of Video Scout. what is described by the metadata and the PP in that they contain references to content information within a TV program. The cen- tral feature of the Segmentation and Indexing module is its ability to integrate information within and/or across the audio, visual, and, transcript domains. Storage and User Interface offers users access and playback to stored programs and stored video clips. In addition, the user interface allows users to modify their PP and their CPP. 3. SYSTEM OVERVIEW These prefer- ences, such as, athlete, movie star, company stock, etc., go beyond Indexing Video Pre-Processing Segmentation Decode Demux Transcript User Input CPPP PPP EPG Storage User Interface Video In Audio Visual Fig. 1. The three modules of Video Scout. Video In gram transcript. We use time-stamps in order to align the transcript data with the related video. Twenty low-level audio parameters are extracted using .wav ﬁles on a PC. The outputs of the Video Pre-Processing are used in the Segmentation and Indexing. 4. VIDEO PRE-PROCESSING in the DAG, such that "% ' "() ", ./ Initial segmentation and indexing is done using closed caption data to divide the video into program and commercial segments. Next the closed captions of the program segments are analyzed for single, double, and triple arrows. Double arrows indicate a speaker change. The system marks text between successive double arrows with a start and end time in order to use it as an atomic closed cap- tions unit. Scout uses these units as the indexing building blocks. In order to determine a segment’s high-level indexing (whether it is ﬁnancial news or a talk show) Scout computes two joint proba- bilities. These are deﬁned as: # ! $ & # ! $ ! + , where # - $ - is a cpd, strictly positive. Now, given the joint probability density function (pdf) # 0 , using the chain rule, we get that # . 1& # $2 + 435---63 # "7 $4 # . According to this equation, the parent set has the property that ! and 8 09 : are conditionally independent given . # ! $ & # ! $ ! + , where # - $ - is a cpd, strictly positive. Now, given the joint probability density function (pdf) # 0 , using the chain rule, we get that . 1 2 456 " 4 # & # $ + 3 - - - 3 # 7 $ # According to this equation, the parent set has the property that ! and 8 09 : are conditionally independent given . p-FIN-TOPIC = p-VTEXT p-KWORDS * p-FACE * p-AUDIO-FIN * p-CC-FIN * p-FACETEXT-FIN (2), In Figure 2 the ﬂow diagram of the BE has the structure of a DAG made up of three layers. In each layer, each element corre- sponds to a node in the DAG. 4. VIDEO PRE-PROCESSING Video pre-processing is done by the Analysis Engine (AE) to com- bine video pre-processing and feature analysis. The AE takes MPEG-2 input and extracts the closed captions while perform- ing a visual analysis for feature extraction. We selected a Philips TriMedia Tricodec card for this task. This card has a TM1000 processor and 8Mb of memory. The TriMedia runs using the hosted mode in a 600 MHz PIII with a WinNT operating system. ”C” code gets downloaded to the TriMedia where analysis is performed. Fig. 2. The three layers of the Segmentation and Indexing module of Video Scout. The AE performs shot detection ﬁrst by extracting a new keyfram when it detects a signiﬁcant difference between sequential I-frames. It employs two DCT based implementations for the frame differ- encing: histogram and macroblock. Unicolor keyframes or frames that appear similar to previously extracted keyframes get ﬁltered out using a one-byte frame signature. This keyframe extraction produces an uncompressed image and a list giving the keyframe number and the probability of a cut. The AE bases this proba- bility on the relative amount above the threshold using the differ- ences between the sequential I-frames. The system then passes keyframes on for videotext detection. The AE looks for video- text using an edge-based method on the uncompressed image and tags each keyframe for the presence or absence of text. These keyframes are also analyzed for the presence of faces. The low-level layer describes signal-processing parameters. In the current implementation parameters include: the visual features color, edge, and shape; twenty audio parameters [7] such as aver- age energy, bandwidth, pitch, mel-frequency cepstral coefﬁcients, linear prediction coding coefﬁcients, and zero-crossings; and the transcript (text pulled from the ASCII characters of the closed- captions). Arrows indicate the combinations of low-level features that create the mid-level features. Mid-level features are associated with whole frames or collections of frames while low-level fea- tures are associated with pixels or short time intervals. Keyframes (ﬁrst frame of a shot), faces, and video text are mid-level visual features; silence, noise, speech, music, speech plus noise, speech plus speech, and speech plus music are mid-level audio features; and keywords and the twenty transcript categories make up the mid-level transcript features. In addition to extracting the visual features, the AE also ex- tracts closed captions from the user data ﬁeld of the MPEG-2 video. 4. VIDEO PRE-PROCESSING The extraction process produces a complete time-stamped pro- High-level features describe semantic video content obtained through the integration of mid-level features across the different modalities. In the current implementation Scout classiﬁes seg- ments as either part of a talk show, ﬁnancial news, or a commercial. layer, twenty closed captions categories are generated: weather, in- ternational, crime, sports, movie, fashion, tech stock, music, auto- mobile, war, economy, energy, stock, violence, ﬁnancial, national (affairs), biotech, disaster, art, and politics. We use a knowledge tree for each category made up of an association table of keywords and categories. After a statistical processing, the system performs categorization using category vote histograms. If a word in the closed captions ﬁle matches a knowledge base keyword, then the corresponding category gets a vote. The probability, for each cat- egory, is given by the ratio between the total number of votes per keyword and the total number of votes for a closed captions para- graph. We based the formal structure of the BE on Bayesian networks [4]. We chose this approach because probabilistic frameworks are designed to deal with uncertain information, and they are appro- priate for representing the integration of information. The BE’s probabilistic integration employs either intra or inter-modalities. Intra-modality integration refers to intergration of features within a single domain. For example: integration of color, edge, and shape information for videotext represents intra-modality integration be- cause it all takes place in the visual domain. Integration of mid- level audio categories with the visual categories face and videotext offers an example of inter-modalities. Video Scout performs segmentation and indexing of TV pro- grams according to the users’ requests. It performs this task by (i) reading users’ requests from the PP and CPP ﬁles, the metadata and the input data; (ii) segmenting the TV program into commer- cial vs. non-commercial parts; (iii) classifying the non-commercial parts into segments based on two high-level categories: ﬁnancial news and talk shows (performed by the BE). Bayesian networks are directed acyclical graphs (DAG) in which the nodes correspond to (stochastic) variables. The arcs describe a direct causal relationship between the linked variables. The strength of these links is given by conditional probability distributions (cpds). More formally, let the set of variables deﬁne a DAG. For each variable there exists a sub-set of variables of , , the parents set of "! , i.e., the predecessors of "! p-TALK-TOPIC = p-VTEXT * p-KWORDS * p-FACE * p-AUDIO-TALK * p-CC- TALK * p-FACETEXT- TALK ( p-TALK-TOPIC = p-VTEXT * p-KWORDS * p-FACE * p-AUDIO-TALK * p-CC- TALK * p-FACETEXT- TALK (3 The audio probabilities p-AUDIO-FIN for ﬁnancial news and p-AUDIO-TALK for talk shows are created by the combination of different individual audio category probabilities. The closed cap- tions probabilities p-CC-FIN for ﬁnancial news and p-CC-TALK for talk shows are chosen as the largest probability out of the list of twenty probabilities. The face and videotext probabilities p- FACETEXT-FIN and p-FACETEXT-TALK are obtained by com- paring the face and videotext probabilities p-FACE and p-TEXT which determine, for each individual closed caption unit, the prob- ability of face and text occurrence. One heuristic used builds on the fact that talk shows are dominated by faces while ﬁnancial news has both faces and text. The high-level indexing is done on each closed captions unit by computing in a new pair of probabilities: p-FIN-TOPIC and p-TALK-TOPIC. The highest value dictates the classiﬁcation of the segment as either ﬁnancial news or talk show. # <>=>? <>=>? !BADCFE <>= + ? < 7 ? 1& # <>=>? !BADCFE $ <>=>? 3 8 # <>= + ? $ <F= + ? # <F= + ? GADC/HJI6E $ <>= + ? GADC/HJI6E 9K 3 8 # 7 $ 7 " # 7 KL $ < 7 ? KL 9M (1) (1) where for each element <>=>? ! there exists a parent set <>=>? ! ; the union of the parent sets for a given level @ , i.e., <>=>? <FNBO2? &QP ADCFE !/R <>=>? ! . There can exist an overlap between the different parent sets for each level. 4. VIDEO PRE-PROCESSING The directed arcs join one node in a given layer with one or more nodes of the preceding layer. Two sets of arcs join the elements of the three layers. For a given layer and for a given element we compute a joint pdf as previously de- scribed. More precisely, for an element (node) ;<>=>? associated with the @ -th layer, the joint pdf is: 9. REFERENCES [1] M. Naphade, T. Huang, ”A Probabilistic Framework for Se- mantic Indexing and Retrieval in Video”, IEEE International Conference on Multimedia and Expo, New York, 31 July-2 August 2000. [2] N. Dimitrova, T. McGee, L. Agnihotri, S. Dagtas, R. Jasin- schi, ”On Selective Video Content Analysis and Filtering,” SPIE Storage and Retrieval for Media Databases, Jan. 2000. Table 3: Financial News Using Audio and Transcript Show CC Units Fin Talk Unclassiﬁed Marketwatch 26 25 0 1 WSW-1 21 17 1 3 WSW-2 27 24 1 2 WSJR 10 9 0 1 Table 4: Talk Shows Using Audio and Transcript Show CC Units Fin Talk Unclassiﬁed Jay Leno 57 4 48 5 Letterman-1 51 18 31 2 Letterman-2 60 18 38 4 Table 3: Financial News Using Audio and Transcript Show CC Units Fin Talk Unclassiﬁed Marketwatch 26 25 0 1 WSW-1 21 17 1 3 WSW-2 27 24 1 2 WSJR 10 9 0 1 Table 4: Talk Shows Using Audio and Transcript Show CC Units Fin Talk Unclassiﬁed Jay Leno 57 4 48 5 Letterman-1 51 18 31 2 Letterman-2 60 18 38 4 [3] www.tivo.com. [4] J. Pearl, ”Probabilistic Reasoning in Intelligent Systems: Networks of Plausible Inference,” Morgan Kaufmann Pub- lishers, Inc., San Mateo, California, 1988. [5] N. Dimitrova, ”Multimedia Content Analysis for Indexing and Retrieval Applications,” ACM Journal of Info. Sci, 1999. [6] N. Vasconcelos and A. Lippman, ”Bayesian Representations and Learning Mechanisms for Content Based Image Re- trieval”, SPIE Storage & Retrieval for Media DB, San Jose, 2000. Tables 5 and 6 display the main results of our integration ef- fort. When we employ all features, Scout can accurately classify both ﬁnancial news and talk shows. Comparing Table 3 to Table 5, Scout loses some precision in classifying ﬁnancial news when all features are combined. However, this slight loss in accuracy is more than made up for in Scout’s increased ability to accurately classify talk shows (Table 4 compared to Table 6). [7] D. Li, I. K. Sethi, N. Dimitrova, and T. McGee, ”Classiﬁ- cation of General Audio Data for Content-Based Retrieval,” Pattern Recognition Letters 2000. [8] T. Syeda-Mahmood and S. Srinivasan, “Detecting Topical Events in Digital Video,” Proc. of ACM Multimedia 2000, Marina Del Rey, November 2000, pp. 85-94. [9] Y. Rui, A. Gupta, and A. Acero, “Automatically Extracting Highlights for TV Baseball Programs,” Proc. 8. CONCLUSION Table 1: Financial News Using Visual and Transcript Show CC Units Fin Talk Unclassiﬁed Marketwatch 26 0 7 19 WSW-1 21 0 16 5 WSW-2 27 0 20 7 WSJR 10 0 5 5 Table 2: Talk Shows Using Visual and Transcript Show CC Units Fin Talk Unclassiﬁed Jay Leno 57 0 46 11 Letterman-1 51 0 44 7 Letterman-2 60 0 53 7 Table 1: Financial News Using Visual and Transcript Show CC Units Fin Talk Unclassiﬁed Marketwatch 26 0 7 19 WSW-1 21 0 16 5 WSW-2 27 0 20 7 WSJR 10 0 5 5 Table 2: Talk Shows Using Visual and Transcript Show CC Units Fin Talk Unclassiﬁed Jay Leno 57 0 46 11 Letterman-1 51 0 44 7 Letterman-2 60 0 53 7 This paper describes our work on Video Scout, a segmentation and indexing system for TV programs. The most important contribu- tions from this work are: (i) integration of information from the audio, visual, and transcript domains; and (ii) high-level classiﬁ- cation of TV program segments based on a probabilistic frame- work. In contrast to the majority of related multimodal integration research, our work on Video Scout comprehensively employs mul- timedia information from all three domains. Experimental results reveal that using audio, visual, and transcript information signif- icantly improves classiﬁcation accuracy across different TV pro- gram genres. Future work includes the introduction of other mul- timedia clues, such as color, and the use of more high-level infor- mation and reasoning. Tables 3 and 4 display results using only audio and transcript information. For this we used the joint probabilities p-FIN-TOPIC = p-AUDIO-FIN * p-KWORDS * p-CC-FIN and p-TALK-TOPIC = p-AUDIO-TALK * p-KWORDS * p-CC- TALK. With the vi- sual features turned off Scout can easily classify ﬁnancial news program, but it has trouble with talk shows. However, ﬁnding talk shows without using visual data is easier than ﬁnding ﬁnan- cial news without using audio data. 6. TOPIC SEGMENTION AND CLASSIFICATION We used Scout to analyze seven TV programs. We looked at the half-hour ﬁnancial news programs Marketwatch, Wall Street Week (WSW), and Wall Street Journal Report (WSJR) as well as the one-hour talk shows hosted by Jay Leno and David Letterman. The total video analyzed was about 6 hours. The results of our experiments are shown in following tables. For each table ”Num. CC Units” indicates the total number of closed captions units for the given TV program. ”Fin. News” and ”Talk Shows” indicate the number of closed captions units (see Section 6 above) classiﬁed as Topic segmentation and classiﬁcation performed by BE is shown in the third layer (high-level) of Figure 2. The complex nature of multimedia content requires integration across multiple domains. We use the comprehensive set of data from the audio, visual, and transcript domains. In the BE structure, Figure 2, for each of the three layers, each node and arrow is associated to a cpd. In the low-level layer the cpd’s are assigned by the AE as described above. For the mid-level Table 6: Talk Shows Using All Features Show CC Units Fin Talk Unclassiﬁed Jay Leno 57 2 50 5 Letterman-1 51 10 41 0 Letterman-2 60 5 53 2 either ﬁnancial news or as a talk show. ”Unclassiﬁed” indicates the number of closed captions units that could not be classiﬁed. In order to better understand the role audio, visual, and tran- script in the integration, we performed experiments using two sub- sets of features. For example: Tables 1 and 2 display results us- ing only visual and transcript information. For this we used the joint probabilities p-FIN-TOPIC = p-VTEXT * p-KWORDS * p- FACE * p-CC-FIN * p-FACETEXT-FIN and p-TALK-TOPIC = p- VTEXT * p-KWORDS * p-FACE * p-CC- TALK * p-FACETEXT- TALK. Using these features, Scout can reasonably classify talk shows, but it cannot ﬁnd ﬁnancial news. Integration is clearly necessary to accurately discriminate be- tween the two program genres. Without using audio, visual, and transcript data it is easy to misclassify entire genres. 9. REFERENCES of ACM Multi- media 2000, Marina Del Rey, November 2000, pp. 105-115. Table 5: Financial News Using All Features Show CC Units Fin Talk Unclassiﬁed Marketwatch 26 24 1 1 WSW-1 21 17 3 1 WSW-2 27 23 4 0 WSJR 10 9 1 0	46,552
https://openalex.org/W2582836438	OpenAlex	Open Science	CC-By	2,014	KENDALA HAKIM DALAM MENCIPTAKAN KEPASTIAN HUKUM, KEADILAN, DAN KEMANFAATAN DI PERADILAN PERDATA	Fence M Wantu	Indonesian	Spoken	7,637	20,733	* Bagian dari disertasi penulis pada tahun 2011 di Fakultas Hukum Universitas Gadjah Mada. ** Alamat korespondensi: fence.wantu@yahoo.co.id KENDALA HAKIM DALAM MENCIPTAKAN KEPASTIAN HUKUM, KEADILAN, DAN KEMANFAATAN DI PERADILAN PERDATA* Fence M. Wantu** Bagian Hukum Acara, Fakultas Hukum Universitas Negeri Gorontalo, Gorontalo Jalan Jenderal Sudirman Nomor 6, Kota Gorontalo, Gorontalo 21752 Abstract The obstacles faced by a judge in realizing the principles of legal certainty, justice, and utility can be categorized into internal and external obstacles. Internal obstacles, which come from within the judge consist of: the appointment of the judge, his education, his understanding of science and knowledge, his morality, and his welfare. On the other hand, external obstacles, which comes from outside the judge, consist of: the independence of judicial power, the formulation of legislations, the prevailing judicial system, community participation, and judge supervision. Keywords: legal certainty, justice, utility. Intisari Kendala yang dihadapi hakim dalam mewujudkan kepastian hukum, keadilan dan kemanfaatan dapat dibedakan menjadi kendala internal dan kendala eksternal. Kendala internal yang datangnya dari dalam diri sendiri. Kendala internal terdiri dari: pengangkatan hakim, pendidikan hakim, penguasaan terhadap ilmu pengetahuan, moral hakim, kesejahteraan hakim. Sementara kendala eksternal datangnya dari luar diri hakim itu sendiri. Kendala eksternal hakim terdiri sebagai berikut, kemandirian kekuasaan kehakiman, pembentukan undang-undang, sistem peradilan yang berlaku, partisipasi masyarakat, pengawasan hakim. Kata Kunci: kepastian hukum, keadilan, kemanfaatan. Kunci: kepastian hukum, keadilan, kemanfaatan. Pokok Muatan A. Latar Belakang Masalah ........................................................................................................... 206 B. Metode Penelitian....................................................................................................................... 207 C. Hasil Penelitian dan Pembahasan............................................................................................... 208 1. Kendala Internal................................................................................................................... 209 2. Kendala Eksternal................................................................................................................. 213 D. Kesimpulan................................................................................................................................. 217 A. Latar Belakang Masalah Salah satu tantangan besar yang terus mem- bayangi perkembangan kekuasaan kehakiman di Indonesia adalah upaya menempatkan peranan hakim yang tepat dalam mewujudkan kepastian hukum, keadilan, dan kemanfaatan, serta me- nempatkan kedudukan interaksinya dengan masyarakat dan negara (adanya hubungan timbal balik). Hakim sebagai salah satu aparat penegak hukum mempunyai tugas sebagai salah satu penentu suatu keputusan perkara dari pihak-pihak yang bersengketa. Agar dapat menyelesaikan masalah atau konflik yang dihadapkan kepada- nya, maka dalam proses pengambilan keputusan, hakim harus mandiri dan bebas dari pengaruh pihak manapun. Hakim dalam mengambil ke- putusan hanya terikat pada peristiwa atau fakta- fakta yang relevan dan kaidah-kaidah hukum yang menjadi atau dijadikan landasan yuridis. Salah satu isu penting untuk menuju masa depan pembangunan hukum termasuk penegakan hukum di Indonesia adalah bagaimana melaksa- nakan kekuasaan kehakiman sesuai dengan tujuan UUD 1945 dan Undang-undang Nomor 48 Tahun 2009 tentang Kekuasaan Kehakiman. Mewujud- kan penegakan hukum di bidang kekuasaan kehakiman yang bebas, merdeka dan mandiri merupakan salah satu tujuan yang ingin dicapai dalam kerangka negara hukum dan demokrasi. Undang-Undang Dasar Negara Republik Indone- sia Tahun 1945 serta Undang-Undang Nomor 4 Tahun 2004 yang sekarang sudah diganti dengan Undang-Undang Nomor 48 Tahun 2009 tentang Kekuasaan Kehakiman telah memberikan dasar pijakan bagi kekuasaan kehakiman untuk me- negakkan keadilan. Namun fakta hukum umum- nya menunjukkan adanya ketidakpercayaan ma- syarakat1 pada kekuasaan kehakiman dikarena- kan salah satu faktor utamanya adalah putusan hakim yang belum mencerminkan nilai keadilan yang didambakan para pencari keadilan.2 Mewujudkan putusan hakim yang didasarkan pada kepastian hukum, keadilan, dan kemanfaatan memang tidak mudah, apalagi tuntutan keadilan, sebab konsep keadilan dalam putusan hakim tidak mudah mencari tolok ukurnya. Adil bagi satu pihak, belum tentu dirasakan sama oleh pihak lain. Hal ini didasarkan pada hakikat keadilan sendiri. Pernyataan tersebut sesuai dengan pendapat dari Sudikno Mertokusumo4 yang menyatakan, “Hakikat keadilan adalah suatu pernilaian dari seseorang kepada orang lain, yang umumnya dilihat dari pihak yang menerima perlakuan saja”. Menghadirkan kepastian hukum, keadilan, dan kemanfaatan dalam suatu putusan hakim sekaligus saat ini sangat sulit. Terdapat beberapa kendala yang harus dihadapi oleh hakim pada saat melaksanakan tugas memeriksa dan memutuskan perkara. Berdasarkan uraian tersebut akhirnya penulis memilih judul tulisan ini. Pokok Muatan MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 206 Sudikno Mertokusumo, 2007, Mengenal Hukum: Suatu Pengantar, Liberty, Yogyakarta, hlm. 77. , mbaga Kajian dan Advokasi Independensi Peradilan, 1999, Menuju Independensi Peradilan, ICEL, Jakarta, hlm. 12-75. 1 Fence M. Wantu, 2011, Peranan Hakim dalam Mewujudkan Kepastian Hukum, Keadilan, dan Kemanfaatan di Peradilan Perdata, Disertasi, Program Doktor Fakultas Hukum, Universitas Gadjah Mada, Yogyakarta. 2 Bandingkan dengan pendapat Kanter, 2001, Etika Profesi Hukum: Sebuah Pendekatan Sosio-Religius, Cet. Pertama, Storia Grafika, Jakarta, hlm. 161. 3 Lembaga Kajian dan Advokasi Independensi Peradilan, 1999, Menuju Independensi Peradilan, ICEL, Jakarta, hlm. 12-75. 4 Sudikno Mertokusumo 2007 Mengenal Hukum: Suatu Pengantar Liberty Yogyakarta hlm 77 Lembaga Kajian dan Advokasi Independensi Peradilan, 1999, Menuju Independensi Peradilan, ICEL, Ja 4 Sudikno Mertokusumo, 2007, Mengenal Hukum: Suatu Pengantar, Liberty, Yogyakarta, hlm. 77. Disertasi, Program Doktor Fakultas Hukum, Universitas Gadjah Mada, Yogyakarta. Bandingkan dengan pendapat Kanter, 2001, Etika Profesi Hukum: Sebuah Pendekatan Sosio-Religius, Cet. Pertama, Storia Jakarta, hlm. 161. L b K ji d Ad k i I d d i P dil 1999 M j I d d i P dil ICEL J k hl 12 75 A. Latar Belakang Masalah Bertitik tolak dari uraian latar belakang di atas, yang dijadikan rumusan masalah, yakni kendala-kendala apa Menyangkut reformasi kekuasaan kehakim- an di Indonesia, Lembaga Kajian dan Advokasi Independensi Peradilan di Indonesia pada tahun 1999 menegaskan3 bahwa yang harus menjadi inti dari reformasi di bidang kekuasaan kehakiman adalah sebagai berikut: Pertama, mewujudkan ke- kuasaan kehakiman sebagai sebuah institusi yang independen; kedua, mengembalikan fungsi hakiki dari kekuasaan kehakiman untuk mewujud- kan keadilan dan kepastian hukum; ketiga, men- jalankan fungsi check and balances bagi institusi kenegaraan lainnya; keempat, mendorong dan memfasilitasi serta menegakkan prinsip-prinsip negara hukum yang demokratis guna mewujudkan kedaulatan rakyat; kelima, melindungi martabat manusia dalam bentuk yang paling konkret. Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 207 yang dihadapi hakim dalam menciptakan ke- pastian hukum, keadilan dan kemanfaatan di peradilan perdata. sampel responden masyarakat pencari keadilan di masing-masing lokasi penelitian yakni berjumlah 3 (tiga) orang. Dengan demikian total responden masyarakat pencari keadilan berjumlah 15 (lima belas) orang. Selanjutnya juga yang dijadikan narasumber adalah para advokat dan para ahli hukum di masing-masing lokasi penelitian. Untuk advokat yakni 3 (tiga) orang advokat di masing-masing lokasi penelitian sehingga jumlah keseluruhan 12 (dua belas) orang advokat. Untuk para ahli hukum yakni 3 (tiga) orang ahli hukum di masing-masing lokasi penelitian sehingga jumlah keseluruhan 12 (dua belas) orang ahli hukum. 8 Pieter Mahmud Marzuki, 2006, Penelitian Hukum, Cetakan Kedua, Prenada Media Group, Jakarta, hlm. 141. 5 Sugeng Istanto, 2007, Penelitian Hukum, Ganda, Yogyakarta, hlm. 29. 6 Soerjono Soekanto, 1986, Pengantar Penelitian Hukum, UI Press, Jakarta, hlm. 52. 7 Sudikno Mertokusumo, Op.cit., hlm. 30. 8 Pieter Mahmud Marzuki, 2006, Penelitian Hukum, Cetakan Kedua, Prenada Media Group, Jakarta, hlm. 141. j , , g 7 Sudikno Mertokusumo, Op.cit., hlm. 30. 5 Sugeng Istanto, 2007, Penelitian Hukum, Ganda, Yogyakarta, hlm. 29. ertokusumo, Op.cit., hlm. 30. mud Marzuki, 2006, Penelitian Hukum, Cetakan Kedua, Prenada Media Group, Jakarta, hlm. 141. B. Metode Penelitian Penelitian hukum adalah penelitian yang membantu pengembangan ilmu hukum dalam mengungkapkan suatu kebenaran hukum. Kon- sekuensinya untuk melakukan penelitian hukum, seseorang harus memahami penelitian itu sendiri dan memahami ilmu hukum.5 Penelitian ini merupakan penelitian yang bersifat yuridis normatif yaitu penelitian hukum yang bertujuan mencari kaidah, norma atau das sollen. Penelitian ini disebut penelitian hukum normatif yang ber- sifat deskriptif karena dimaksudkan untuk memberikan gambaran yang rinci tentang fokus yang diteliti dengan memanfaatkan norma-norma hukum yang ada, sehingga dapat menjawab permasalahan yang diteliti, namun demikian penelitian ini juga membutuhkan data lapangan. Adapun data yang diperoleh dari penelitian lapangan adalah data primer yang diperoleh langsung dari subjek penelitian yakni narasumber dan responden. Jumlah semua populasi yang ada di Pengadilan Negeri dan Pengadilan Tinggi berdasarkan lokasi penelitian ini adalah 119 (seratus sembilan belas) orang hakim PN dan hakim PT, yang terdiri dari hakim tinggi berjumlah 66 (enam puluh enam) orang dan hakim negeri berjumlah 53 (lima puluh tiga) orang. Selanjutnya yang diambil sampel sebagai responden di masing-masing Pengadilan Negeri dan Pengadilan Tinggi terdiri dari 3 (tiga) orang hakim dan hakim tinggi. Jumlah responden hakim dan hakim tinggi dari Pengadilan Negeri dan Pengadilan Tinggi adalah 27 (dua puluh tujuh) orang. Untuk populasi masyarakat pencari keadilan didasarkan pada perkara perdata yang diteliti di masing- masing lokasi penelitian. Kemudian ditentukan Penelitian ini mendasarkan pada keter- sediaan bahan hukum sekunder. Bahan hukum sekunder merupakan bahan yang memberikan penjelasan mengenai bahan hukum primer, seperti rancangan undang-undang, hasil-hasil penelitian, hasil karya dari kalangan hukum.6 Berkenaan dengan hal tersebut Sudikno Mertokusumo7 menyatakan dalam upaya menyempurnakan data (bahan hukum) yang diperoleh dari penelitian kepustakaan, dapat dilakukan penelitian lapangan. Menurut Marzuki bahan hukum merupakan dokumen-dokumen resmi berupa semua publikasi tentang hukum. Publikasi tentang hukum meliputi peraturan perundang-undangan, peraturan pe- merintah, buku-buku teks, kamus-kamus hukum, jurnal-jurnal hukum, dan komentar-komentar atas putusan pengadilan.8 Bahan hukum tersebut kemudian dibedakan dalam tiga kelompok bagian yakni bahan hukum primer, bahan hukum sekunder, dan bahan hukum tersier. Sunaryati Hartono menyatakan bahan hukum primer dibagi lagi dalam mandatory sources, yaitu perundang-undangan nasional yang dikeluarkan oleh penguasa dari wilayah hukum sendiri, dan persuasive primary sources, yaitu perundang-undangan dari provinsi yang berbeda (tetapi yang menyangkut hal yang sama) atau MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 208 Mahkamah Agung dianggap merisaukan pen- dukung kebebasan pers dan demokrasi. B. Metode Penelitian Putusan Kasasi dengan perkara MA Nomor 3215 K/Pdt/ 2001 diputus pada tanggal 28 Agustus 2007 dan baru dibacakan secara terbuka pada 30 Agustus 2007 oleh Ketua Muda Mahkamah Agung Bi- dang Militer German Hoediarto (Ketua Majelis), M. Taufiq (Anggota), dan Baharuddin Qaudry (Anggota). Putusan Mahkamah Agung tersebut telah mengabaikan semangat kebebasan pers yang merupakan tonggak demokrasi sebagai salah satu wujud kedaulatan rakyat sebagaimana dijamin dan diatur dalam Pasal 28F UUD 1945 dan Un- dang-Undang Nomor 40 Tahun 1999 tentang Pers.12 putusan pengadilan dari wilayah hukum yang berbeda.9 Kemudian bahan hukum sekunder yaitu bahan pustaka yang berisikan informasi tentang bahan hukum primer. Selanjutnya bahan hukum tersier merupakan bahan pustaka yang berisikan bahan hukum yang memberi petunjuk terhadap bahan hukum primer dan bahan hukum sekunder. Sarana yang digunakan untuk mengumpulkan bahan hukum sekunder adalah studi dokumen. Metode analisis yang dipergunakan dalam penelitian ini adalah analisis kualitatif. Metode analisis kualitatif merupakan tata cara penelitian yang menghasilkan data deskriptif. Hal ini didasarkan pendapat Maria S.W. Sumardjono,10 yang menyatakan dalam penelitian hukum normatif yang mempergunakan data sekunder, penelitiannya pada umumnya bersifat deskriptif atau deskriptif-eksploratif serta analisisnya bersifat kualitatif. Menurut Gijssels dan van Hoecke, analisis data dilakukan dalam tiga cara yaitu: Pertama, sistematisasi data (cara deskriptif); kedua, penjelasan (cara eksplikatif); ketiga, perbaikan dan pembaharuan (cara preskriptif atau normatif).11 Perkara lain yang perlu dikemukakan yakni gugatan yang diajukan oleh PT Bumijaya Sentosa kepada PT Mitra Bangun Griya yang akhirnya melalui putusan Pengadilan Jakarta Selatan Perkara Nomor 63/Pdt.G/2004/PN.Jak.Sel, yang memenangkan PT Bumijaya Sentosa, tetapi tidak dapat dieksekusi. PT Bumijaya Sentosa belum dapat memperoleh keadilan yang sudah didapatkannya tersebut.13 Selain itu perkara putusan Pengadilan Negeri Jakarta Selatan14 antara David Nusa Widjaya dan Tarunodjojo Nusa (Penggugat) melawan Badan Penyehatan Perbankan Nasional (Tergugat). Majelis Eksaminasi yang terdiri dari kalangan akademisi hukum, praktisi hukum, dan aktivis Lembaga Swadaya Masyarakat antara lain menyimpulkan dalam suatu negara hukum yang demokratis, tuntutan kekuasaan kehakiman yang merdeka, berwibawa, bersih, dan jujur, harus dapat diwujudkan. Keprihatinan yang mendalam tentunya melihat reformasi hukum yang masih berjalan lambat dan belum memberikan rasa keadilan bagi masyarakat. Sejumlah masalah 9 Sunaryati Hartono, 2006, Penelitian Hukum di Indonesia pada Akhir Abad ke-20, Cetakan Kedua, Alumni, Bandung, hlm. 134. 9 Sunaryati Hartono, 2006, Penelitian Hukum di Indonesia pada Akhir Abad ke-20, Cetakan Kedua, Alumni, Bandung, hlm. 134. 10 Maria S.W. Sumardjono, 2001, Pedoman Pembuatan Usulan Penelitian Sebuah Panduan Dasar, Cetakan Ketiga, Gramedia Pustaka Utama, Jakarta, hlm. 10. 11 Jan Gijssels dan Mark van Hoecke, 2000, Apakah Teori Hukum itu? (Terj. Arief Sidharta), Laboratorium Hukum Fakultas Hukum Universitas Katholik Parahyangan, Bandung. 12 Ibid. 13 Lihat dalam Majalah Pledoi, No. 10, Vol. 1, 2007, hlm. 26. 14 Putusan Pengadilan Negeri Jakarta Selatan Nomor 303/Pdt.G/2003/Pn.Jak.Sel. dalam perkara Ng Tjuen Wie alias David Nusa Widjaya dan Tarunodjojo Nusa Melawan BPPN. j , , , , 14 Putusan Pengadilan Negeri Jakarta Selatan Nomor 303/Pdt.G/2003/Pn.Jak.Sel. dalam perkara Ng Tjuen Wie alias David Nu dan Tarunodjojo Nusa Melawan BPPN. ssels dan Mark van Hoecke, 2000, Apakah Teori Hukum itu? (Terj. Arief Sidharta), Laboratorium Hukum Fakultas Hukum sitas Katholik Parahyangan, Bandung. naryati Hartono, 2006, Penelitian Hukum di Indonesia pada Akhir Abad ke 20, Cetakan Kedua, Alumni, Bandung, hlm. 134. aria S.W. Sumardjono, 2001, Pedoman Pembuatan Usulan Penelitian Sebuah Panduan Dasar, Cetakan Ketiga, Gramedia Pusta ama, Jakarta, hlm. 10. 13 Lihat dalam Majalah Pledoi, No. 10, Vol. 1, 2007, hlm. 26. 1. Kendala Internal Pada dasarnya kendala internal merupakan kendala yang mempengaruhi hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan yang datangnya dari dalam diri, dan berkaitan dengan sumber daya manusia (SDM) hakim itu sendiri. Pada dasarnya kendala internal hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan, berdasarkan hasil temuan yang diadakan adalah terdiri dari sebagai berikut: Banyak yang beranggapan bahwa rekrut- men hakim belum didasarkan pada norma- norma profesionalisme atau kemampuan pribadi hakim yang bersangkutan, yang pada akhirnya mengakibatkan terjadinya penyimpangan-penyimpangan di dalam proses peradilan yang melahirkan putusan hakim yang kurang mencerminkan kepasti- an hukum dan rasa keadilan masyarakat.18 Padahal dalam rekrutmen hakim perlu dipersyaratkan adanya intelektualitas (yang terkait dengan kemampuan penguasaan hukum materiil, hukum formil dan penemuan hukum secara tepat dan benar); integritas (kejujuran); pendidikan, penataran, refresh- ing, rapat-rapat berkala, dan diklat; dan langkah-langkah efisiensi dan efektifitas kelas-kelas diklat.19 C. Hasil Penelitian dan Pembahasan Suatu gejala yang mengkhawatirkan di bidang penegakan hukum dan keadilan di pengadilan adalah keadilan hukum yang tidak sejalan lagi dengan keadilan masyarakat. Dampak langsung dari kejadian ini adalah hilangnya kepercayaan masyarakat kepada negara dan timbulnya inisiatif masyarakat untuk membuat pengadilan sendiri yang sering berdampak pada tindakan main hakim sendiri. Ada lagi perkara gugatan pencemaran nama baik mantan Presiden Soeharto kepada majalah Time yang diputuskan kasasinya oleh Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 209 yang layak mendapat perhatian berkenaan dengan bidang hukum antara lain: sistem peradilan yang dipandang kurang independen dan imparsial; belum memadainya perangkat hukum yang mencerminkan keadilan sosial; inkonsistensi dalam penegakan hukum; masih adanya intervensi terhadap hukum; lemahnya perlindungan hukum terhadap masyarakat; rendahnya kontrol secara komprehensif terhadap penegakan hukum; dan belum meratanya tingkat keprofesionalan para penegak hukum. Berkaitan dengan hal tersebut, dapat dilihat bahwa terdapat kendala-kendala hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan. Kendala-kendala tersebut, pada garis besarnya dapat dibedakan dalam kendala internal dan kendala eksternal. hakim, yaitu: adanya integritas, kecakapan, dan kualifikasi calon hakim; metode seleksi hakim harus memberikan perlindungan bagi pengangkatan hakim dari motivasi-motivasi yang tidak layak; dan tidak boleh ada diskriminasi terhadap calon hakim. Dalam konteks hukum di Indonesia pengangkatan hakim mengacu pada peraturan perundang- undangan yang berlaku antara lain Undang- undang tentang Mahkamah Agung;15 Undang-Undang tentang Peradilan Umum;16 dan Undang-Undang tentang Peradilan Tata Usaha Negara.17 Pada dasarnya persyaratan untuk dapat diangkat menjadi hakim, baik dalam berbagai lingkungan peradilan maupun Mahkamah Agung adalah sama. Letak perbedaannya hanya pada usia dan pengalaman. 15 Undang-Undang Nomor 3 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung jis. Undang-Undang Nomor 5 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung, Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. 16 Undang-Undang Nomor 49 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 2 Tahun 1986 jis. Undang-Undang Nomor 8 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum, Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. 17 17 Undang-Undang Nomor 51 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 5 Tahun 1986 jis. Undang-Undang Nomor 9 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara, Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara. 19 Tim Peneliti Fakultas Hukum Universitas Gadjah Mada, 2003, Rekrutmen dan Karir di Bidang Peradilan, Hasil Peneliti Hukum Nasional, Jakarta. 18 Pendapat sebagian dari narasumber yang terdiri dari para ahli hukum di masing-masing lokasi dan sebagian narasumber advokat. Data hasil wawancara Oktober – November 2008, Januari 2009, Juli 2009, November 2009, Januari 2010. 19 Tim Peneliti Fakultas Hukum Universitas Gadjah Mada, 2003, Rekrutmen dan Karir di Bidang Peradilan, Hasil Penelitian, Komisi Hukum Nasional, Jakarta. g g 18 Pendapat sebagian dari narasumber yang terdiri dari para ahli hukum di masing-masing lokasi dan sebagian narasumber adv hasil wawancara Oktober – November 2008, Januari 2009, Juli 2009, November 2009, Januari 2010. 8 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum, Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. 17 Undang-Undang Nomor 51 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 5 Tahun 1986 jis. Undang-Undang Nomor 9 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara, Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara. 18 Pendapat sebagian dari narasumber yang terdiri dari para ahli hukum di masing masing lokasi dan sebagian narasumber advokat Data g g 18 Pendapat sebagian dari narasumber yang terdiri dari para ahli hukum di masing-masing lokasi dan sebagian narasumber advokat. Data hasil wawancara Oktober – November 2008, Januari 2009, Juli 2009, November 2009, Januari 2010. 19 Tim Peneliti Fakultas Hukum Universitas Gadjah Mada, 2003, Rekrutmen dan Karir di Bidang Peradilan, Hasil Penelitian, Komisi Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. 16 Undang-Undang Nomor 49 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 2 Tahun 1986 jis. Undang-Undang Nomor 8 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum, Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. 17 Undang-Undang Nomor 51 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 5 Tahun 1986 jis. Undang-Undang Nomor a) Pengangkatan Hakim Salah satu ketentuan hukum interna- sional yang menentukan persyaratan peng- angkatan hakim adalah Pasal 10 Prinsip- Prinsip Dasar Independensi Kehakiman (Basic Principles on the Independence of the Judiciary). Ketentuan hukum internasional mengandung beberapa hal yang harus diperhatikan dalam proses pengangkatan Salah satu ketentuan hukum interna- sional yang menentukan persyaratan peng- angkatan hakim adalah Pasal 10 Prinsip- Prinsip Dasar Independensi Kehakiman (Basic Principles on the Independence of the Judiciary). Ketentuan hukum internasional mengandung beberapa hal yang harus diperhatikan dalam proses pengangkatan MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 210 bersangkutan masih di perguruan tinggi. Khusus untuk perekrutan dari sarjana baru, haruslah ditentukan lebih dahulu sistem penataran atau training, oleh sebab itu perlu dipersiapkan buku training yang baik dan inovatif; (5) untuk penempatan hakim magang perlu ditentukan di pengadilan yang relatif dekat dengan tempat asal calon peserta sehingga dapat meringankan beban ekonomisnya dan tidak ditentukan secara acak; (6) perekrutan hakim non-karir untuk tingkat pertama dan tinggi dirasa masih perlu dipikirkan dan direkomendasikan. Untuk mendapatkan hakim yang baik perlu dikaji mulai dari perekrutan hakim sampai dengan perjalanan karir hakim. Berdasarkan hasil wawancara20 dengan responden ditemukan di lapangan terdapat kelemahan dalam sistem rekrutmen/peng- angkatan hakim yang antara lain disebabkan oleh sebagai berikut: (1) sistem rekrutmen/ pengangkatan hakim yang masih cenderung tertutup dan kurang berorientasi untuk men- dapatkan sumber daya hakim yang baik; (2) adanya rekrutmen/pengangkatan hakim yang mempunyai indikasi adanya kolusi, korupsi, dan nepotisme; (3) kurang pembinaan dan pelatihan terhadap hakim yang ada terutama bagi yang masih berstatus calon hakim; (4) mekanisme dan promosi jabatan hakim tidak jelas; (5) jumlah pengadilan dan jumlah hakim yang tidak sesuai dengan kebutuhan, apalagi jika dihubungkan dengan jumlah perkara yang harus ditanganinya; (6) kurang- nya mekanisme pengawasan yang tegas terhadap para hakim; (7) belum optimalnya pemberian penghargaan (reward) kepada para hakim yang berprestasi; dan (8) tidak tegasnya penerapan sanksi hukuman ter- hadap kinerja hakim yang bermasalah. Kebijakan yang dapat diambil mengenai pengangkatan hakim, yakni sebagai berikut: Pertama, pembaharuan sistem rekrutmen dan karir hakim yang berlaku selama ini, khususnya untuk calon-calon hakim men- datang sehingga pada gilirannya dapat diwujudkan hakim yang layak, profesional, berpengetahuan luas dan tinggi, berdedikasi dan bermoral, jujur, adil, terbebas dari kolusi, korupsi, dan nepotisme. Diusulkan kepada Presiden dan DPR untuk meninjau ulang pranata hukum yang ada dan sekaligus mem- bentuk pranata hukum yang baru berkaitan dengan sistem rekrutmen dan karir hakim yang layak. 20 Pendapat sebagian dari narasumber yang terdiri dari para ahli hukum di masing-masing lokasi dan sebagian narasumber advokat. Data hasil wawancara Oktober – November 2008, Januari 2009, Juli 2009, November 2009, Januari 2010. Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 211 b) Pendidikan Hakim pengadilan negeri menunjukkan bahwa tingkat pendidikan formal hakim masih tertinggal.22 Memang diakui bahwa tingkat pendidikan hakim tidak selamanya menjadi ukuran untuk menilai kualitas putusan, tapi paling tidak dengan hakim mengikuti pen- didikan setingkat magister, maka bekal pengetahuan seperti penemuan hukum, teori, dan filsafat hukum yang diberikan pada pen- didikan setingkat magister dengan metode yang berbeda sewaktu diberikan pada tingkat pendidikan sarjana menjadi modal berarti bagi hakim. Mengingat beratnya tanggung jawab, maka hakim haruslah terseleksi dari pen- didikan yang berkualitas, berbudi pekerti luhur, mempunyai dedikasi tinggi. Sebagai penegak hukum dan keadilan, hakim harus dapat berdiri tegak dan mandiri dalam mem- berikan keadilan. Keadilan yang diberikan adalah demi keadilan berdasarkan ketuhanan yang maha esa, yang hanya dapat dilakukan oleh mereka yang terpilih dan terpanggil, yakni mereka yang benar-benar mempunyai panggilan jiwa dan hati nurani. Pendidikan dan pelatihan (diklat) merupakan bagian integral dari sistem pembinaan karir bagi hakim. Pendidikan dan pelatihan hakim yang diselenggarakan dalam bentuk klasikal mampu menghasilkan lulusan hakim yang memiliki kualitas yang tinggi, namun jang- kauan terhadap pesertanya terbatas pada jumlah peserta yang hadir dalam kelas yang bersangkutan. 21 Ketentuan tersebut dapat dilihat dalam Cetak Biru Mahkamah Agung yang diterbitkan pada Tahun 2003, yang sekarang sudah diperbaharui dengan Cetak Biru Mahkamah Agung Tahun 2008. 22 Data yang diperoleh di PN Tilamuta dan PN Limboto pada bulan November 2008 dan bulan Januari 2009. 23 Pasal 5 ayat (1) Undang-Undang 48 Tahun 2009. a) Pengangkatan Hakim Kedua, seleksi ulang untuk pengadaan hakim yang layak, profesional, berpengetahuan luas dan tinggi, berdedikasi dan bermoral, jujur, adil, terbebas kolusi, korupsi, dan nepotisme dari kalangan hakim yang ada selama ini. Diajukan usul kepada Presiden dan DPR untuk mengambil ke- bijakan seleksi ulang terhadap para hakim yang ada, yang lulus ditempatkan kembali sebagai hakim, sedang yang tidak lulus dipensiunkan dini atau dipekerjakan sebagai non-hakim. Mengantisipasi kebutuhan ke depan, maka untuk perbaikan rekrutmen hakim, perlu diperhatikan hal-hal sebagai berikut: (1) pimpinan yang merekrut haruslah mem- punyai kemampuan organisasi, tidak cukup hanya mempunyai kemampuan ilmu pengetahuan hukum, oleh sebab itu perlu ditetapkan sistem rekrutmen yang baik dan terencana; (2) sistem rekrutmen hakim harus diubah, dan perlu dipikirkan adanya ke- tentuan yang tegas, siapa yang akan direkrut dan siapa yang akan menjadi tim perekrut serta siapa yang akan menjadi pengawasnya; (3) orang yang melamar seharusnya sudah tahu apakah tugas dan kewajiban hakim. Dalam rekrutmen, faktor mental atau moral lebih diutamakan; (4) perlu dipikirkan pere- krutan hakim mulai dilakukan sejak yang g g g 22 Data yang diperoleh di PN Tilamuta dan PN Limboto pada bulan November 2008 dan bulan Januari 2009. 23 P l 5 (1) U d U d 48 T h 2009 23 Pasal 5 ayat (1) Undang-Undang 48 Tahun 2009. 22 Data yang diperoleh di PN Tilamuta dan PN Limboto pada bulan November 2008 dan bulan Januari 2009. 23 Pasal 5 ayat (1) Undang-Undang 48 Tahun 2009. 23 Pasal 5 ayat (1) Undang-Undang 48 Tahun 2009. 21 Ketentuan tersebut dapat dilihat dalam Cetak Biru Mahkamah Agung yang diterbitkan pada Tahun 2003, yang sekarang sudah d dengan Cetak Biru Mahkamah Agung Tahun 2008. 24 Contoh dapat dilihat pada Putusan PN Jakarta Selatan No. 1466/Pdt.G/2006/pn.Jak.Sel. yaitu gugatan wanprestasi Surat Pengakuan Utang. Bahwa putusan majelis hakim yang memeriksa perkara ini sebagian besar tidak didasarkan pada ketentuan yang ada, memang dalam beberapa bagian putusan diakui sudah memberikan perlindungan keadilan bagi para tergugat misalnya ditolaknya permohonan putusan verzet dan biaya jasa pengacara yang digunakan penggugat harus ditanggung oleh tergugat. Namun demikian secara umum putusan ini tidak memberikan kepastian hukum, keadilan dan kemanfaatan. Kepastian hukum yang dimaksudkan di sini adalah putusan hakim tersebut tidak memberikan jalan keluar terhadap permasalahan hukum yang dihadapi oleh para pihak, putusan tersebut tidak sesuai dengan tujuan undang-undang yang dijadikan dasar dari putusan hakim, tidak mengandung aspek stabilitas yaitu dapat memberikan rasa tertib dan aman bagi masyarakat. Sementara unsur keadilan tidak dipenuhi dalam kasus ini karena putusan hakim tidak mengakui persamaan antara hak dan kewajiban, tidak adanya kesesuaian penerapan antara peraturan perundang-undangan dengan rumusan putusan hakim, tidak mengandung keadilan dan ketidakseimbangan dalam masyarakat. Selanjutnya tidak terpenuhi kriteria kemanfaatan dalam putusan hakim karena putusan ini tidak memberikan kepuasan bagi para pihak, munculnya polemik baru bagi pihak-pihak yang berperkara, tidak adanya keseimbangan dalam masyarakat. 25 Laporan Akhir Komisi Hukum Nasional Republik Indonesia tentang Standar Disiplin Profesi Tahun 2003. c) Penguasaan terhadap Ilmu Hukum Idealnya hakim harus menguasai per- kembangan ilmu hukum. Penguasaan terha- dap ilmu hukum merupakan hal yang tidak dapat diabaikan oleh hakim dalam menjalan- kan tugasnya sehari-hari. Hal ini berkaitan juga dengan pelaksanaan seleksi calon hakim, yaitu untuk mencegah lolosnya calon hakim yang tidak mempunyai kualitas dalam penguasaan ilmu hukum. Untuk mencapai hal itulah hakim harus menguasai teori-teori hukum yang bersifat teknis, seperti interpre- tasi dan konstruksi hukum yang pada prin- sipnya memberikan ruang gerak kepada para hakim untuk menemukan hukum pada suatu kasus yang diperiksa dan memberikan motivasi kepada hakim untuk tidak terpaku kepada bunyi ketentuan pasal-pasal mati suatu aturan hukum. Bahkan dalam konteks memutuskan suatu perkara oleh hakim, dike- nal pula contra legem, yaitu mekanisme yang membolehkan hakim menyimpangi suatu ketentuan yang nyata-nyata bertentangan dengan rasa keadilan masyarakat. Tindakan seperti itu secara yuridis telah pula mendapat legitimasi Undang-Undang 48 Tahun 200923 yang pada prinsipnya mengamanatkan Pembaruan sistem pendidikan dan pe- latihan hakim merupakan bagian yang tidak terpisahkan dengan Cetak Biru Mahkamah Agung. Dalam Cetak Biru Mahkamah Agung diuraikan berbagai program pembaruan sistem pendidikan dan pelatihan hakim yang meliputi: 21 (1) model sistem pendidikan dan pelatihan yang sesuai bagi hakim di Indo- nesia; (2) lembaga pendidikan dan pelatih- an; (3) tenaga instruktur dan metode sistem pendidikan dan pelatihan; (4) penyusunan program dan kurikulum sistem pendidikan dan pelatihan; dan (5) distribusi kesempatan dan pemanfaatan hasil sistem pendidikan dan pelatihan. Data yang diperoleh di lapangan khusus- nya di Provinsi Gorontalo sebagian besar pengadilan baik pengadilan tinggi maupun MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 212 kepada hakim untuk menggali nilai-nilai hukum yang ada di masyarakat agar putusan yang dibuat dapat memenuhi rasa keadilan masyarakat yang ada. dalam bentuk-bentuk penafsiran, bahkan menciptakan hukum baru melalui putusan- putusannya. d) Moral Hakim Putusan hakim yang berkualitas menun- tut adanya perpaduan antara ilmu pengeta- huan (knowledge) dalam energi mental, energi emosional, dan energi spiritual. Optimalisasi penguasaan ilmu pengetahu- an dalam energi-energi tersebut akan menyentuh akal, perasaan, dan keyakinan, sehingga tidak heran banyak yang menyata- kan putusan hakim harus didasarkan pada keyakinan hakim. Berdasarkan data hasil penelitian24 ditemukan berbagai kontroversi dalam putusan dan pertimbangan hukum yang tidak mendasarkan pada fakta yang ada dan tidak sesuai dengan hukum yang berlaku, merupakan kekurangan utama dalam pengetahuan hakim. Demikian juga dalam memutuskan perkara hakim hanya berpedoman pada undang-undang semata tanpa melihat perkembangan yang ada. Padahal begitu banyak peraturan-peratur- an undang-undang di bidang kekuasaan kehakiman yang sudah dilengkapi oleh Mahkamah Agung melalui surat edaran MA, surat keputusan MA, yurisprudensi, dan surat keputusan bersama dengan lembaga lain. Sudah saatnya paradigma lama yang menyatakan bahwa hakim hanya sekedar mulut atau corong undang-undang, harus segera diubah dengan paradigma bahwa hakim mampu memberi makna melalui penemuan hukum atau konstruksi hukum Hakim harus mempunyai kemampuan profesional serta moral dan integritas yang tinggi agar mampu mencerminkan rasa keadilan, memberikan manfaat dan kepastian hukum. Selain itu hakim harus mempunyai kemampuan untuk berkomunikasi serta menjalankan peranan dan statusnya yang dapat diterima oleh masyarakat, hakim juga harus mempunyai iman dan taqwa yang baik. Selain itu, tanggung jawab hakim berat karena harus bertanggung jawab kepada Tuhan Yang Maha Esa, diri sendiri, para pihak, masyarakat, pengadilan yang lebih tinggi dan ilmu pengetahuan hukum. d) Moral Hakim Sebagaimana dalam ketentuan laporan akhir Komisi Hukum Nasional Republik Indonesia tahun 200325 dinyatakan bahwa sebagai pengemban profesi hukum selalu dituntut pengembangan dirinya senantiasa didasarkan pada nilai-nilai moralitas umum (common morality) yang terdiri sebagai berikut: (1) nilai-nilai kemanusiaan (humanity), dalam arti penghormatan pada keluhuran martabat kemanusiaan; (2) nilai-nilai keadilan (justice), dalam arti dorongan untuk selalu memberi- kan kepada orang apa yang menjadi haknya; (3) nilai-nilai kepatutan atau kewajaran, dalam arti bahwa upaya mewujudkan keadilan dalam masyarakat selalu diwarnai oleh kesadaran untuk selalu memperhatikan dan memperhitungkan rasionalitas situasi dan Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 213 rasa keadilan individual anggota masyarakat; (4) nilai-nilai kejujuran, dalam arti dorongan kuat untuk selalu memelihara kejujuran dan penghindaran diri dari perbuatan- perbuatan yang curang; (5) keharusan untuk memiliki kualitas keahlian dan keilmuan yang tinggi dalam disiplin ilmu hukum pada para pengembannya; (6) kesadaran untuk selalu menghormati dan menjaga integritas dan kehormatan profesinya; (7) nilai-nilai pelayanan dan kepentingan publik, dalam pengertian bahwa di dalam pengembangan profesi hukum telah melekat semangat keberpihakan pada hak-hak dan kepuasan masyarakat pencari keadilan yang merupakan konsekuensi langsung dari dipegang teguhnya nilai-nilai keadilan, kejujuran, dan kredibilitas profesional dan keilmuan. kehidupan hakim di daerah-daerah sesuai dengan kenyataan yang ada tidak sama dengan hakim yang ada di kota-kota besar.27 Menurut Soekotjo Soeparto Koordinator Bidang Hubungan Antar Lembaga Komisi Yudisial, perlu segera dilakukan peningkatan kesejahteraan hakim agar dapat memacu kinerja. Perbaikan terhadap kesejahteraan hakim sudah saatnya mendapat perhatian yang lebih serius.28 Idealnya penegakan hukum yang me- nyeluruh juga semestinya mencakup upaya pemerintah untuk memperbaiki nasib para hakim. Sebab tidak sedikit hakim yang memang hanya hidup dari kejujuran dan kesederhanaan. Hanya dengan berbekal nurani, para hakim itu bekerja dengan kondisi seadanya, di tengah godaan materi yang mudah saja didapatkan. Banyaknya godaan yang menghadang para hakim tersebut semoga tidak membuat idealisme mereka menjadi goyah. Komitmen para hakim untuk bekerja profesional sesuai dengan kode etika dan sumpah profesi adalah bagian dari darma bakti mereka terhadap negara. Pada dasarnya kepribadian moral yang kuat, yang harus dimiliki oleh hakim tidak lain adalah kejujuran; mampu menjadi diri sendiri; bertanggung jawab; memiliki kemandirian moral; memiliki keberanian moral; dan memiliki kerendahan hati. 2. Kendala Eksternal Kendala eksternal adalah kendala yang mempengaruhi hakim dalam menciptakan ke- pastian hukum, keadilan dan kemanfaatan yang datangnya dari luar diri hakim itu sendiri. Hal ini berkaitan dengan sistem peradilan dan penegakan hukumnya. Pada dasarnya kendala eksternal hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan, berdasarkan hasil temuan yang diadakan adalah terdiri dari sebagai berikut: p daksi, “Mendorong Terwujudnya Kekuasaan Kehakiman yang Merdeka”, Buletin Komisi Yudisial, Vol. III, No. 2, Oktober 2008. enelitian yang diperoleh di salah satu pengadilan negeri di daerah, di mana terdapat hakim-hakim yang bertugas di daerah masih di rumah-rumah penduduk. 26 Hasil wawancara dengan responden hakim di daerah seperti PN Tilamuta yang kurang mendapat perhatian. Hasil wawa diperoleh ini tidak sesuai dibandingkan dengan tunjangan hakim yang diberikan negara begitu tinggi. e) Kesejahteraan Hakim 2. Kerja para hakim menuntut perhatian ekstra keras, namun ternyata sampai saat ini tidak mendapat perhatian serius pemerin- tah.26 Hakim secara proporsional harus dihargai, sehingga bagi hakim yang mem- punyai prestasi baik dapat diberikan peng- hargaan sesuai dengan apa yang telah dilakukannya. Bentuk penghargaan tersebut dapat berupa mutasi dan promosi jabatan yang lebih tinggi. Hasil penelitian melalui wawancara dengan hakim yang tinggal di daerah seperti PN Tilamuta Provinsi Goron- talo menunjukkan selain jaminan sosial berupa perumahan yang dijadikan tempat kediaman para hakim masih kurang, 26 Hasil wawancara dengan responden hakim di daerah seperti PN Tilamuta yang kurang mendapat perhatian. Hasil wawancara yang diperoleh ini tidak sesuai dibandingkan dengan tunjangan hakim yang diberikan negara begitu tinggi. 27 Hasil penelitian yang diperoleh di salah satu pengadilan negeri di daerah, di mana terdapat hakim-hakim yang bertugas di daerah masih tinggal di rumah-rumah penduduk. gg p 28 Tim Redaksi, “Mendorong Terwujudnya Kekuasaan Kehakiman yang Merdeka”, Buletin Komisi Yudisial, Vol. III, N diperoleh ini tidak sesuai dibandingkan dengan tunjangan hakim yang diberikan negara begitu tinggi. 27 Hasil penelitian yang diperoleh di salah satu pengadilan negeri di daerah, di mana terdapat hakim-hakim yang bertugas d tinggal di rumah-rumah penduduk. b) Pembentukan Undang-Undang Melihat kenyataan saat ini, kondisi peraturan hukum acara perdata begitu memprihatinkan. Kendatipun kemerdekaan Indonesia sudah lebih dari 65 tahun, ternyata hukum acara perdata masih menggunakan produk peninggalan pemerintah Hindia Belanda yaitu H.I.R. dan R.Bg. Peraturan yang mengatur hukum acara perdata, dibandingkan dengan peraturan lainnya jelas terlambat dan tertinggal.32 Peraturan hukum acara perdata sampai saat ini belum ada unifikasi, masih bersifat pluralistik yaitu tersebar dalam berbagai peraturan yang ada. Hal ini disebabkan masih kurangnya kemauan politik dari pemerintah dan lembaga legislatif untuk membuat peraturan tersendiri tentang hukum acara perdata. Salah satu langkah strategi yang dapat dilakukan dalam menciptakan putusan hakim yang mencerminkan kepastian hukum, keadilan dan kemanfaatan adalah dengan melakukan reformasi terhadap berbagai peraturan yang berkaitan dengan kekuasaan kehakiman pada umumnya dan hukum acara perdata pada khususnya. Pelaksanaan kemandirian kekuasaan kehakiman dapat dibedakan menjadi tiga macam, yaitu kemandirian lembaganya, ke- mandirian proses peradilan, dan kemandirian hakim itu sendiri.30 Terkait dengan intervensi atau campur tangan atas kemandirian hakim dalam proses peradilan perlu kiranya di- kemukakan apa yang pernah menjadi penga- laman salah seorang mantan hakim yakni Sahlan Said yang menyatakan pada saat melaksanakan tugas proses peradilan pernah diintervensi oleh pimpinannya, karena tidak mau melaksanakan perintah tersebut akhir- nya beliau dikucilkan dengan berbagai sindiran-sindiran di lingkungan pengadilan tersebut.31 Untuk menyikapi demikian, akan ber- gantung pada hati nurani hakim itu sendiri. Hakim dalam posisi ini dituntut untuk men- junjung tinggi idealismenya dengan mem- a) Kemandirian Kekuasaan Kehakiman Sasaran penyelenggaraan kekuasaan kehakiman adalah untuk menumbuhkan ke- mandirian dalam rangka mewujudkan per- MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 214 adilan yang berkualitas. Penyelenggaraan kekuasaan kehakiman dilakukan dengan meningkatkan integritas dan ilmu penge- tahuan. Kemandirian kekuasaan kehakiman merupakan asas yang sifatnya universal. Hal ini dapat berarti dalam melaksanakan proses peradilan, hakim pada dasarnya bebas dalam memeriksa dan mengadili perkara dan bebas dari campur tangan kekuasaan ekstra yudisial. Pada asasnya hakim itu mandiri, tetapi kemandirian hakim tersebut tidaklah bersifat mutlak. Hal ini dikarena- kan dalam menjalankan tugasnya hakim secara mikro dibatasi oleh Pancasila, UUD, peraturan perundang-undangan, kehendak para pihak, ketertiban umum dan kesusilaan. Selain itu dalam menjalankan tugasnya, secara makro kemandirian hakim dibatasi oleh sistem pemerintahan, sistem politik, sistem ekonomi.29 pertahankan kebebasan dan kemandiriannya. Menurut penulis, modal dasar yang harus dimiliki oleh seorang hakim dalam menjaga kemandiriannya perlu ditunjang juga dengan keahlian yang memadai. Hakim yang mampu mempertahankan kemandiriannya akan dapat berfungsi sebagai penegak hukum yang baik dalam menjalankan tugas dan wewenangnya. Kemandirian ini membawa hakim selalu berpedoman pada prinsip-prinsip hukum yang adil dan dapat dipertanggungjawabkan. Sudikno Mertokusumo, 2009, Hukum Acara Perdata Indonesia, Liberty,Yogyakarta. 30 Bambang Sutiyoso dan Puspitasari, 2005, Aspek-Aspek Perkembangan Kekuasaan Kehakiman di Indonesia, UII Press, Yogyakarta, hlm. 52. ang diperoleh melalui salah satu acara yang ada di Metro TV yakni wawancara pada tanggal 17 Januari 2010 dengan Sahlan Said. o Mertokusumo, Loc.cit. 29 Sudikno Mertokusumo, 2009, Hukum Acara Perdata Indonesia, Liberty,Yogyakarta. 29 Sudikno Mertokusumo, 2009, Hukum Acara Perdata Indonesia, Liberty,Yogyakarta. 30 Bambang Sutiyoso dan Puspitasari, 2005, Aspek-Aspek Perkembangan Kekuasaan Kehakiman di Indonesia, UII Press, Yogyakarta, hlm. 52. , , y, gy ng Sutiyoso dan Puspitasari, 2005, Aspek-Aspek Perkembangan Kekuasaan Kehakiman di Indonesia, UII Press, Yogyakarta, 2 32 Sudikno Mertokusumo, Loc.cit. o Mertokusumo, 2009, Hukum Acara Perdata Indonesia, Liberty,Yogyakarta. y gy 30 Bambang Sutiyoso dan Puspitasari, 2005, Aspek-Aspek Perkembangan Kekuasaan Kehakiman di In hlm. 52. 29 Sudikno Mertokusumo, 2009, Hukum Acara Perdata Indonesia, Liberty,Yogyakarta. 30 Bambang Sutiyoso dan Puspitasari, 2005, Aspek-Aspek Perkembangan Kekuasaan Kehakiman di Indonesia, UII Press, Yogyakarta, hlm. 52. 31 Data yang diperoleh melalui salah satu acara yang ada di Metro TV yakni wawancara pada tanggal 17 Januari 2010 dengan Sahlan Said. 32 S dik M t k L it e) Sistem Pengawasan Hakim Doner menyatakan paling tidak ada tiga macam pengawasan hakim. Pertama, pengawasan hukum, yaitu suatu bentuk yang ditujukan untuk mengetahui apakah wewe- nang sudah dilaksanakan sesuai dengan ketentuan hukum yang berlaku. Kedua, pengawasan administrasi, yaitu suatu bentuk pengawasan yang bertujuan untuk mengukur efisiensi kerja. Ketiga, pengawasan politik, yaitu suatu bentuk pengawasan yang diguna- kan untuk mengukur segi-segi kemanfaat- an.34 Berkaitan dengan pengawasan perilaku hakim, pada dasarnya benar jika para hakim harus diawasi oleh pimpinan dan juga oleh lembaga independen yang mendapat kewenangan konstitusional untuk itu. Pentingnya pembaharuan sistem per- adilan, karena dengan usaha tersebut akan melahirkan produk-produk putusan pengadil- an yang berkualitas. Pada akhirnya putusan pengadilan yang berkualitas menjadi sumber hukum yang dapat dipakai dalam kehidup- an berbangsa dan bernegara. Artinya putusan pengadilan tersebut dapat dipertang- gungjawabkan dan diikuti secara sukarela bagi para pihak, dan tidak melahirkan masalah baru di tengah-tengah masyarakat. Terlepas dari belum sempurnanya sistem peradilan kita, upaya penegakan hukum terhadap hukum acara terutama hukum acara perdata dan nilai peradilan harus tetap dijaga dan dipertahankan oleh semua pihak terutama oleh para aparatur pengadilan itu sendiri. Pengawasan internal terhadap hakim dilakukan oleh Mahkamah Agung sebagai- mana diatur oleh Undang-Undang Ke- kuasaan Kehakiman dan Undang-undang Mahkamah Agung. Pengawasan terhadap hakim oleh Mahkamah Agung dapat dibagi menjadi dua, yakni pengawasan terhadap hakim agung yang berada di lingkungan MA dan pengawasan terhadap hakim di pengadilan tingkat pertama dan tingkat banding. Sementara pengawasan eksternal terhadap perilaku hakim yang dilakukan oleh Komisi Yudisial diharapkan dapat menutupi kelemahan pengawasan internal yang di- lakukan oleh MA. c) Sistem Peradilan yang Berlaku Pada dasarnya pembaharuan sistem peradilan menyangkut beragam aspek, mulai dari aspek pembenahan sumber daya manusia Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 215 yakni peningkatan kualitas hakim dan pembenahan aspek administrasi peradilan. Peningkatan kualitas hakim dapat berarti putusan hakim yang jujur, adil dan mengikuti perkembangan hukum hanya akan lahir dari sosok pribadi yang baik dan mempunyai ilmu pengetahuan hukum. Gambaran ideal- nya hakim tersebut hanya lahir jika di- tunjang dengan sistem administrasi keuang- an lembaga pengadilan dan politik hukum yang kondusif. kesadaran partisipasi masyarakat sangat diharapkan. Kondisi adanya kesadaran partisipasi masyarakat sangat mendukung. Sebaliknya jika kesadaran partisipasi masyarakat tidak ada dan rendah, maka apa yang diharapkan tidak terwujud. 34 Lihat dalam Komisi Yudisial, Vol. 1, No. 5, April 2007, hlm. 18. 33 Lihat dalam Varia Peradilan, No. 244, Maret 2006. d) Partisipasi Masyarakat Dalam pembenahan penegakan hukum penting untuk diintensifkan partisipasi publik/masyarakat. Hal ini didasarkan pada alasan bawa masyarakat mempunyai peran dalam mengawasi jalannya peradilan seba- gaimana disebutkan dalam undang-undang kekuasaan kehakiman. Partisipasi masyarakat tidak sekedar melibatkan lembaga swadaya masyarakat (LSM), tetapi para individu yang ada dalam masyarakat.33 Untuk menciptakan putusan hakim yang mencerminkan kepastian hukum, keadilan dan kemanfaatan, maka Tuntutan terhadap pengawasan eksternal dan internal supaya benar-benar dijalankan, pada akhirnya melahirkan Surat Keputusan Bersama antara Ketua Mahkamah Agung MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 216 RI dan Ketua Komisi Yudisial RI No. 047/ KMA/SK/IV/2009 dan No. 02/SKB/P.KY/ IV/2009 tentang Kode Etik dan Pedoman Perilaku Hakim.35 Pada dasarnya prinsip- prinsip dasar Kode Etik dan Pedoman Peri- laku Hakim diimplementasikan dalam sepuluh aturan perilaku sebagai berikut, berperilaku adil, berperilaku jujur, berperi- laku arif dan bijaksana, bersikap mandiri, berintegritas tinggi, bertanggung jawab, menjunjung tinggi harga diri, berdisiplin tinggi, berperilaku rendah hati, dan bersikap profesional. rekrutmen yang benar-benar mempunyai kualitas tidak didasarkan pada kolusi, korupsi, dan nepotisme. Kedua, peningkatan nilai- nilai moral dan etika. Ketiga, pendidikan dan pelatihan hakim tetap dilakukan secara rutinitas. Peningkatan kesejahteraan hakim dan jaminan keselamatan hakim dan ke- luarganya; (2) Perlu merumuskan putusan hakim yang lebih mendekatkan pada nilai- nilai keadilan, kepastian hukum dan keman- faatan. Alasan dan pertimbangan hukum di dalam putusan hakim dapat diterima terutama oleh para pihak dan masyarakat pada umumnya; (3) Peningkatan penegakan hukum dengan menyelesaikan perkara- perkara perdata di pengadilan dengan bertitik tolak pada asas hukum dan asas peradilan yang baik. Asas hukum harus dijadikan landasan utama hakim dalam menyelesaikan dan memutuskan perkara; (4) Partisipasi masyarakat dalam penegakan hukum ter- utama di pengadilan; (5) Meningkatkan pendidikan kesadaran masyarakat terhadap proses hukum beracara di pengadilan. 35 Lihat dalam Varia Peradilan, No. 283, Juni 2009. f) Upaya Mengatasi Kendala Hakim Kendala yang dihadapi harus diakui dan diterima oleh komunitas hukum sebagai problem yang tidak secara eksklusif dapat diselesaikan hanya dengan pendekatan ilmu hukum saja. Problem tersebut harus dicari- kan solusi dengan pendekatan multi disiplin ilmu pengetahuan. Beberapa konsep yang perlu diwujudkan dalam mengatasi kendala internal dan eksternal, antara lain: (1) peng- gunaan hukum yang berkeadilan; (2) adanya lembaga pengadilan yang independen, bebas dan tidak memihak; (3) aparatur penegak hukum (hakim) yang profesional; (4) pe- negakan hukum yang berdasarkan prinsip keadilan; (5) partisipasi publik; dan (6) ada- nya kontrol melalui eksaminasi yang efektif. Langkah yang menjadi perhatian yang dapat dilakukan hakim dalam menerima, memeriksa dan memutus perkara di pengadilan, sehingga putusan hakim dapat mencerminkan kepastian hukum, keadilan dan kemanfaatan di peradilan perdata, adalah sebagai berikut: (1) Meletakkan setiap kasus yang ditanganinya dengan cara memaparkan setiap kasus dalam sebuah ikhtisar. Dalam hal ini hakim memaparkan secara singkat duduk perkara dari sebuah kasus; (2) Menerjemahkan atau mengkualifikasi setiap kasus tersebut dari peristiwa abstrak ke dalam peristiwa hukum atau konkret; (3) Menyeleksi aturan-aturan hukum yang tepat untuk dijadikan sebagai dasar hukum terhadap peristiwa konkret tersebut; (4) Melakukan analisis dan menafsir- kan terhadap aturan-aturan hukum tersebut yang telah dijadikan sebagai dasar hukum dalam Selain itu, dalam rangka melakukan reformasi hukum khususnya untuk mengatasi kendala internal dan eksternal hakim ada beberapa hal yang harus dilakukan antara lain: (1) Penataan kembali struktur dan lembaga kekuasaan kehakiman yang ada, termasuk rekrutmen sumber daya manusia- nya yang berkualitas. Kemandirian kekuasa- an kehakiman harus ada jaminan dan tetap dapat diwujudkan dalam proses peradilan di pengadilan. Hal yang perlu diperhatikan juga adalah: Pertama, soal pengangkatan/ Wantu, Kendala Hakim dalam Menciptakan Kepastian Hukum, Keadilan, dan Kemanfaatan 217 memecahkan kasusnya; (5) Menerapkan aturan hukum tersebut dalam kasusnya. kekuasaan kehakiman harus ada jaminan dan tetap dapat diwujudkan dalam proses peradilan di pengadilan. Hal yang perlu diperhatikan juga adalah: (1) soal pengangkatan/rekrutmen yang benar-benar mempunyai kualitas tidak didasarkan pada KKN; (2) peningkatan nilai- nilai moral dan etika; (3) pendidikan dan pelatihan hakim tetap dilakukan secara rutin; (4) peningkatan kesejahteraan hakim dan jaminan keselamatan hakim dan keluarganya. Kedua, perlu merumuskan putusan-putusan hakim yang lebih mendekatkan pada nilai-nilai keadilan, kepastian hukum dan kemanfaatan. Alasan dan pertimbangan hukum di dalam putusan hakim dapat diterima terutama oleh para pihak dan masyarakat pada umumnya. Ketiga, peningkatan penegakan hukum dengan menyelesaikan perkara-perkara perdata di pengadilan dengan bertitik tolak pada asas hukum dan asas peradilan yang baik. D. Kesimpulan Pada dasarnya kendala yang dihadapi hakim dalam mewujudkan kepastian hukum, keadilan dan kemanfaatan dapat dibedakan dalam kendala internal dan kendala eksternal. Pada dasarnya kendala internal hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan, berdasarkan hasil temuan yang diadakan adalah terdiri dari pengangkatan hakim, pendidikan hakim, penguasaan terhadap ilmu pengetahuan, moral hakim, dan kesejahteraan hakim. Sementara kendala eksternal hakim dalam menciptakan kepastian hukum, keadilan dan kemanfaatan adalah kemandirian kekuasaan kehakiman, pembentukan undang-undang, sistem peradilan yang berlaku, partisipasi masyarakat, dan sistem pengawasan hakim. Untuk mengatasi kendala internal dan eksternal hakim ada beberapa hal yang harus dilakukan antara lain: Pertama, penataan kem- bali struktur dan lembaga kekuasaan kehakim- an yang ada, termasuk rekrutmen sumber daya manusianya yang berkualitas. Kemandirian f) Upaya Mengatasi Kendala Hakim Asas hukum harus dijadikan landasan utama hakim dalam menyelesaikan dan memutuskan perkara. Keempat, partisipasi masyarakat dalam penegakan hukum terutama di pengadilan. Kelima, meningkatkan pendidikan kesadaran masyarakat terhadap proses hukum beracara di pengadilan. Daftar Pustaka Lembaga Kajian dan Advokasi Independensi Peradilan, 1999, Menuju Independensi Peradilan, ICEL, Jakarta. E. Peraturan Perundang-undangan g g Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. B. Artikel Jurnal Undang-Undang Nomor 8 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 2 Tahun 1986 tentang Peradilan Umum. Komisi Yudisial, “Mendorong Terwujudnya Kekuasaan Kehakiman yang Merdeka”, Buletin Komisi Yudisial, Vol. III, No. 2, Oktober 2008. Undang-Undang Nomor 9 Tahun 2004 tentang Per- ubahan Atas Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara. Komisi Yudisial, Vol. 1, No. 5, April 2007. A. Buku Gijssels, Jan dan Mark van Hoecke, 2000, Apakah Teori Hukum itu?, (Terj. Arief Sidharta), Laboratorium Hukum Fakultas Hukum Universitas Katholik Parahyangan, Bandung. Marzuki, Pieter Mahmud, 2006, Penelitian Hukum, Cetakan Kedua, Prenada Media Group, Jakarta. Hartono, Sunaryati, 2006, Penelitian Hukum di Indonesia pada Akhir Abad ke-20, Cetakan Kedua, Alumni, Bandung. Mertokusumo, Sudikno, 2007, Mengenal Hukum: Suatu Pengantar, Liberty, Yogyakarta. _____________, 2009, Hukum Acara Perdata Indonesia, Liberty, Yogyakarta. Istanto, Sugeng, 2007, Penelitian Hukum, Ganda, Yogyakarta. Sumardjono, Maria, S.W., 2001, Pedoman Pem- buatan Usulan Penelitian Sebuah Panduan Dasar, Cetakan Ketiga, Gramedia Pustaka Utama, Jakarta. Kanter, 2001, Etika Profesi Hukum: Sebuah Pendekatan Sosio-Religius, Cetakan Pertama, Storia Grafika, Jakarta. Laporan Akhir Komisi Hukum Nasional Republik Indonesia tentang Standar Disiplin Profesi Tahun 2003. Laporan Akhir Komisi Hukum Nasional Republik Indonesia tentang Standar Disiplin Profesi Tahun 2003. Utama, Jakarta. Soekanto, Soerjono, 19 Hukum, UI Press, J Soekanto, Soerjono, 1986, Pengantar Penelitian Hukum, UI Press, Jakarta. Soekanto, Soerjono, 1986, Pengantar Penelitian Hukum, UI Press, Jakarta. MIMBAR HUKUM Volume 25, Nomor 2, Juni 2013, Halaman 205 - 218 218 Sutiyoso, Bambang dan Puspitasari, 2005, Aspek- Aspek Perkembangan Kekuasaan Kehakiman di Indonesia, UII Press, Yogyakarta. Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara. Undang-Undang Nomor 5 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. Undang-Undang Nomor 5 Tahun 1986 tentang Peradilan Tata Usaha Negara. Undang-Undang Nomor 5 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. Undang-Undang Nomor 5 Tahun 2004 tentang Perubahan Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. F. Putusan Pengadilan Putusan Pengadilan Negeri Jakarta Selatan Nomor 303/Pdt.G/2003/Pn.Jak.Sel. peri- hal dalam perkara Ng. Tjuen Wie alias David Nusa Widjaya dan Tarunodjojo Nusa Me- lawan BPPN, 2003. Varia Peradilan, No. 244, Maret 2006. Varia Peradilan, No. 283, Juni 2009. C. Hasil Penelitian/Tugas Akhir Undang-Undang Nomor 3 Tahun 2009 tentang Per- ubahan Kedua Atas Undang-Undang Nomor 14 Tahun 1985 tentang Mahkamah Agung. Fakultas Hukum Universitas Gadjah Mada, 2003, Rekrutmen dan Karir di Bidang Peradilan, Hasil Penelitian, Komisi Hukum Nasional, Jakarta. Undang-Undang 48 Tahun 2009 tentang Kekuasaan Kehakiman. Undang-Undang Nomor 49 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 2 Tahun 1986. Wantu, Fence, M., 2011, Peranan Hakim Dalam Mewujudkan Kepastian Hukum, Keadilan, dan Kemanfaatan di Peradilan Perdata, Disertasi, Program Doktor Fakultas Hukum, Universitas Gadjah Mada, Yogyakarta. Undang-Undang Nomor 51 Tahun 2009 tentang Perubahan Kedua Atas Undang-Undang Nomor 5 Tahun 1986. D. Artikel Majalah atau Koran Majalah Pledoi, No. 10, Vol. 1, 2007. Varia Peradilan, No. 244, Maret 2006. Varia Peradilan, No. 283, Juni 2009.	34,449
16/hal-ephe.archives-ouvertes.fr-hal-02306650-document.txt_3	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	2,527	5,699	Mitte, Kawahara, Yen, Cummings & Zwick, 2015 Clade Eulepidoptera Kiriakoff, 1948 Clade Heteroneura Tillyard, 1918 Clade Coelolepida Nielsen & Kristensen, 1996 Clade Glossata Fabricius, 1775 Clade Neoptera Martynov, 1923 Infra-classe Pterygota Brauer, 1885 Classe Hexapoda Blainville, 1816 Clade Ecdysozoa Aguinaldo, Turbeville, Linford, Rivera, Garey, Raff & Lake, 1997 Infra-Règne Protostomia Grobben, 1908 90 2. Phénologie Période de vol période de vol estimée O données non-Françaises X données Françaises X nouvelles données Françaises 25/4 30/4 5/5 10/5 15/5 20/5 25/5 30/5 5/6 10/6 15/6 20/6 25/6 30/6 5/7 10/7 15/7 20/7 25/7 30/7 5/8 10/8 15/8 20/8 25/8 30/8 5/9 10/9 Sesia apiformis Sesia bembeciformis X Sesia melanocephala X X Paranthrene tabaniformis X Paranthrene ssp synagriformis X Paranthrene insolitus Chamaesphecia mysiniformis Chamaesphecia aerifrons Chamaesphecia osmiaeformis Chamaesphecia ramburi Chamaesphecia dumonti Chamaesphecia nigrifrons Chamaesphecia bibioniformis Chamaesphecia anthraciformis Chamaesphecia palustris Chamaesphecia euceraeformis Chamaesphecia leucopsiformis Chamaesphecia empiformis Chamaesphecia tenthrediniformis Pyropteron chrysidiformis Pyropteron triannuliformis Pyropteron meriaeformis Pyropteron hispanica Pyropteron muscaeformis Pyropteron leucomelaena Pyropteron affinis Bembecia ichneumoniformis Bembecia albanensis X X X X X X X X X X Endémique de Corse X X X X O X X X X O X X X X X X X X X X X X X X Endémique de Corse O X X X X X X X X X X X X X X X X X X X X X X X X X X X X X O X X Période de vol 25/4 30/4 5/5 10/5 15/5 20/5 25/5 30/5 5/6 10/6 15/6 20/6 25/6 30/6 5/7 10/7 15/7 20/7 25/7 30/7 5/8 10/8 15/8 20/8 25/8 30/8 5/9 10/9 Bembecia psoraleae Bembecia fibigeri Bembecia scopigera X X Bembecia iberica X X Bembecia megillaeformis Bembecia astragali Bembecia himmighoffeni Bembecia uroceriformis Synanthedon theryi Synanthedon scoliaeformis Synanthedon mesiaeformis Synanthedon spheciformis Synanthedon stomoxiformis X Synanthedon culiciformis X Synanthedon formicaeformis Synanthedon flaviventris Synanthedon andrenaeformis Synanthedon soffneri Synanthedon melliniformis Synanthedon myopaeformis Synanthedon vespiformis Synanthedon codeti Synanthedon conopiformis Synanthedon tipuliformis Synanthedon spuleri Synanthedon loranthi Synanthedon cephiformis Tinthia tineiformis Pennisetia hylaeiformis X X X X X X X X Heure française d'attirance par les phéromones de Wageningen Sesia apiformis Sesia bembeciformis Sesia melanocephala Paranthrene tabaniformis Paranthrene insolitus Chamaesphecia mysiniformis Chamaesphecia aerifrons Chamaesphecia osmiaeformis Chamaesphecia ramburi Chamaesphecia dumonti Chamaesphecia nigrifrons 10:00 10:30 X X 11:00 11:30 12:00 12:30 13:00 13:30 X X X X X 14:00 X X X X X X X 14:30 15:00 15:30 16:30 17:00 17:30 18:00 18:30 19:00 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X 16:00 X X X X X X X X X X X X X X X X X meriaeformis X X X hispanica X X X X X X X X X anthraciformis Chamaesphecia palustris Chamaesphecia euceraeformis Chamaesphecia leucopsiformis Chamaesphecia empiformis Chamaesphecia tenthrediniformis Pyropteron chrysidiformis Pyropteron triannuliformis Pyropteron Pyropteron Pyropteron muscaeformis X X X Chamaesphecia X X X X X X X X X bibioniformis 20:00 X X Chamaesphecia X X X X X X X Pyropteron leucomelaena Pyropteron affinis X X Bembecia ichneumoniformis X X X X X X X X Bembecia albanensis X X X X X X X X Bembecia psoraleae X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X 20:30 21:00 21:30 Heure française d'attirance par les phéromones de Wageningen 10:00 10:30 11:00 11:30 12:00 12:30 X X X X X X X X X X X X X X X 13:00 13:30 14:00 14:30 Bembecia scopigera Bembecia iberica Bembecia megillaeformis Bembecia astragali Bembecia himmighoffeni Bembecia uroceriformis X X Synanthedon theryi X X X X Synanthedon scoliaeformis X X X X Synanthedon mesiaeformis Synanthedon spheciformis X X X Synanthedon stomoxiformis X X X Synanthedon culiciformis X X X X X X X 16:00 16:30 X 17:00 Synanthedon flaviventris Synanthedon andrenaeformis X X X Synanthedon soffneri X X X Synanthedon melliniformis Synanthedon myopaeformis X X X Synanthedon vespiformis Synanthedon codeti Synanthedon conopiformis Synanthedon tipuliformis Synanthedon spuleri Synanthedon loranthi Synanthedon cephiformis Tinthia tineiformis Pennisetia hylaeiformis 17:30 18:00 X X X X X X X X X X X X X X X X X X X X X 19:00 19:30 20:00 20:30 X X X X X X X X X X X X 94 21:00 X X X 18:30 X X formicaeformis X X X Synanthedon X 15:30 X fibigeri X 15:00 X Bembecia X X X X X X X X X 21:30 Phéromones PHEROMONES WAGENINGEN + Efficacité de la phéromone sur l'espèce Mélanges de phéromones Espèce B.albanensis B. astragali BEIC ++ faible optimum meria: api+myo+vesp PEHY SYTI + SYFO SYCO SEAP SYMY PATA ++++ + SYVE SYSCO SYSO SYCU ++ ++++ B.scopigera SYSP SYPO SYSE aeri euce leuco meria ++ + + + + + + + + + + Coronilla minima,Ononis natrix,L.corniculatus Fabacées sp ++ + + ++ ++ + +++ + + + ++ +++ ++ +++ + ++ + ++++ + + + ++ Euphorbia sp +++ Euphorbia seguieriana et sp ++ ++ ++++ + ++ + ++ Thymus sp, Mentha sp, Origanum vulgare +++ Euphorbia sp ++ C. leucopsiformis + Stachys sp + C. euceraeformis C. mysiniformis Bituminaria bituminosa Ulex sp et Spartium junceum + C. dumonti Fabacées sp Genista tinctoria ++ C.anthraciformis Astragalus et Colutea sp Ononis fructicosa et rotundifolia ++++ +++ plantes hôtes Ononis sp +++ +++ ++ B.uroceriformis C.osmiaeformis SYSPU ++ + C.nigrifrons SYFL ++ + ++ C.empiformis ++++ +++ B. psoraleae C. bibioniformis élevé leuco: api+hyl+myo B. megillaeformis C.aerifrons +++ eucera: myo+vesp ++ B. himmighoffeni B.ichneumoniformis moyen aeri: api+hyl+vesp B. fibigeri B. iberica ++ ++ ++ +++ + Marrubium sp, Stachys sp Hypericum perforatum ++++ Salvia sp C. palustris Euphorbia palustris hyl + api + C.tenthrediniformis +++ + P.hylaeiformis P.affinis + P.chrysidiformis + tab ++ tip ves + + ++++ + ++++ + + ++++ + ++ ++ + +++ + + ++ P.leucomelaena ++ + + P.meriaeformis ++ P. triannuliformis culi cono ++ + P.hispanica + soff flavi spul ++++ euce leuco meria plantes hôtes ++ +++ Helianthemum sp +++ ++ + + + ++++ ++ ++++ ++ ++ + Populus sp Rubus idaeus ++ Rumex scutatus Poterium minor ++ + Rumex hydrolapathum, R.obtusifolius et sp Rumex acetosa, R.acetosella, R.scutatus Armeria et Dianthus sp Rumex scutatus et R.acetosella Populus sp Salix caprea et sp Populus tremula + S.andrenaeformis + + +++ +++ S.cephiformis S.conopiformis aeri Quercus robur et sp S. melanocephala S. codeti seq ++ + S. bembeciformis pola ++++ ++++ S. : département où l'absence de l'espèce est certaine ou probable : département où la preuve de la détermination est la mise en collection des pièces génitales. : département où la preuve de la détermination est la mise en collection d'un individu. : d épartement où la preuve de la détermination est une photo. Mâle Bembecia albanensis Femelle Bembecia albanensis Mâle Bembecia astragali Femelle Bembecia astragali 98 5. Niveau de validation automatique des données CD_NOM NOM_COMPLET 253298 Bembecia albanensis albanensis (Rebel, 1918) 784241 Bembecia astragali (Joannis, 1909) 784241 Bembecia astragali (Joannis, 1909) 247108 247113 247113 247110 247110 247104 848334 247107 247109 247109 247114 253294 253295 247091 253296 247088 247095 247093 Bembecia fibigeri Z. Laštůvka & A. Laštůvka, 1994 Bembecia himmighoffeni (Staudinger, 1866) Bembecia himmighoffeni (Staudinger, 1866) Bembecia iberica Špatenka, 1992 Bembecia iberica Špatenka, 1992 Bembecia ichneumoniformis (Denis & Schiffermüller, 1775) Bembecia megillaeformis luqueti Špatenka, 1992 Bembecia psoraleae Bartsch & Bettag, 1997 Bembecia scopigera (Scopoli, 1763) Bembecia scopigera (Scopoli, 1763) Bembecia uroceriformis (Treitschke, 1834) Chamaesphecia aerifrons aerifrons (Zeller, 1847) Chamaesphecia aerifrons sardoa (Staudinger, 1856) Chamaesphecia anthraciformis (Rambur, 1832) Chamaesphecia bibioniformis tengyraeformis (Boisduval, 1840) Chamaesphecia dumonti Le Cerf, 1922 Chamaesphecia empiformis (Esper, 1783) Chamaesphecia euceraeformis (Ochsenheimer, 1816) MIXTE DETERMINABILITE PREUVE EXPERT COL_PG EX_CRITERE_DET COMMENTAIRE OBS_PG_LAB Confusion principalement avec B. ichneumoniformis. Confusion des mâles de petites tailles avec B. iberica. L'attraction par la phéromone "meriaeformis" oriente la détermination. MALE DELICATE FEMELLE SIMPLE COL_IND PHOTO FILET FILET MIXTE MALE FEMELLE MALE FEMELLE EXPERT DELICATE SIMPLE DELICATE SIMPLE COL_PG COL_IND PHOTO COL_IND PHOTO OBS_PG_LAB FILET FILET FILET FILET MIXTE MIXTE MIXTE MALE FEMELLE MIXTE MIXTE MIXTE MIXTE EXPERT EXPERT EXPERT EXPERT SIMPLE SIMPLE DELICATE DELICATE SIMPLE COL_PG COL_IND COL_PG COL_PG PHOTO PHOTO PHOTO PHOTO PHOTO OBS_PG_LAB COL_IND OBS_PG_LAB OBS_PG_LAB FILET FILET FILET FILET FILET Confusion principalement avec B. albanensis. Non revue en France depuis 1913. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. MIXTE MIXTE MIXTE MIXTE DELICATE DELICATE DELICATE DELICATE COL_IND COL_IND COL_IND COL_IND FILET FILET FILET COL_IND Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. L'attraction par la phéromone "vespiformis" oriente la détermination. Confusion avec B. albanensis. Confusion avec de petits mâles de B. albanensis. 247094 247084 247089 247086 247092 24 7087 247096 848356 784257 253292 253291 247134 247103 253297 247100 247102 247099 247101 247098 247079 247080 247122 247132 247127 247128 247119 Chamaesphecia leucopsiformis (Esper, 1800) Chamaesphecia mysiniformis (Boisduval, 1840) Chamaesphecia nigrifrons (Le Cerf, 1911) Chamaesphecia osmiaeformis (Herrich-Schäffer, 1848) Chamaesphecia palustris Kautz, 1927 Chamaesphecia ramburi (Staudinger, 1866) Chamaesphecia tenthrediniformis (Denis & Schiffermüller, 1775) Eusphecia melanocephala (Dalman, 1816) Paranthrene insolitus polonica Schnaider, 1939 Paranthrene tabaniformis synagriformis (Rambur, 1866) Paranthrene tabaniformis tabaniformis (Rottemburg, 1775) Pennisetia hylaeiformis (Laspeyres, 1801) Pyropteron affine (Staudinger, 1856) Pyropteron chrysidiforme chrysidiforme (Esper, 1782) Pyropteron hispanicum (Kallies, 1999) Pyropteron leucomelaenum (Zeller, 1847) Pyropteron meriaeforme (Boisduval, 1840) Pyropteron muscaeforme (Esper, 1783) Pyropteron triannuliforme (Freyer, 1843) Sesia apiformis (Clerck, 1759) Sesia bembeciformis (Hübner, 1806) Synanthedon andrenaeformis (Laspeyres, 1801) Synanthedon cephiformis (Ochsenheimer, 1808) Synanthedon codeti (Oberthür, 1881) Synanthedon conopiformis (Esper, 1782) Synanthedon culiciformis (Linnaeus, 1758) MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIX MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE TRES_SIMPLE SIMPLE SIMPLE SIMPLE DELICATE DELICATE DELICATE SIMPLE SIMPLE SIMPLE SIMPLE TRES_SIMPLE DELICATE TRES_SIMPLE DELICATE DELICATE DELICATE DELICATE DELICATE SIMPLE SIMPLE SIMPLE DELICATE SIMPLE DELICATE SIMPLE 100 PHOTO PHOTO PHOTO PHOTO COL_IND COL_IND COL_IND PHOTO PHOTO PHOTO PHOTO PHOTO COL_IND PHOTO PHOTO COL_IND COL_IND COL_IND COL_IND PHOTO PHOTO PHOTO COL_IND PHOTO PHOTO PHOTO FILET FILET FILET FILET COL_IND COL_IND FILET SANS FILET FILET FILET SANS COL_IND SANS FILET COL_IND COL_IND COL_IND COL_IND SANS SANS FILET COL_IND FILET FILET FILET Confusion avec plusieurs espèces. Non revue en France depuis 1930. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Confusion avec plusieurs espèces. Il n'existe qu'une seule citation ancienne pour cette espèce en France. Confusion avec S. loranthi. Confusion avec plusieurs espèces. Confusion possible avec S. myopaeformis. 247121 247120 247131 247124 247116 253300 253301 247115 247123 247117 247130 253299 343146 247129 247126 247133 Synanthedon flaviventris (Staudinger, 1883) Synanthedon formicaeformis (Esper, 1783) Synanthedon loranthi (Králícek, 1966) Synanthedon melliniformis (Laspeyres, 1801) Synanthedon mesiaeformis (Herrich-Schäffer, 1846) Synanthedon myopaeformis myopaeformis (Borkhausen, 1789) Synanthedon myopaeformis typhiaeformis (Borkhausen, 1789) Synanthedon scoliaeformis (Borkhausen, 1789) Synanthedon soffneri Špatenka, 1983 Synanthedon spheciformis (Denis & Schiffermüller, 1775) Synanthedon spuleri (Fuchs, 1908) Synanthedon stomoxiformis stomoxiformis (Hübner, 1790) Synanthedon theryi Le Cerf, 1916 Synanthedon tipuliformis (Clerck, 1759) Synanthedon vespiformis (Linnaeus, 1761) Tinthia tineiformis (Esper, 1789) MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE MIXTE SIMPLE TRES_SIMPLE DELICATE DELICATE TRES_SIMPLE SIMPLE TRES_SIMPLE SIMPLE SIMPLE SIMPLE DELICATE SIMPLE_SCBG SIMPLE DELICATE SIMPLE TRES_SIMPLE 101 PHOTO PHOTO COL_IND PHOTO PHOTO PHOTO PHOTO PHOTO PHOTO PHOTO COL_IND PHOTO PHOTO PHOTO PHOTO PHOTO FILET SANS COL_IND FILET SANS FILET SANS FILET FILET FILET COL_IND FILET FILET FILET FILET SANS Confusion avec S. cephiformis Confusion avec S. vespiformis Confusion possible avec S. culiciformis Confusion avec plusieurs espèces. Confusion possible avec S. myopaeformis dans le sud de la France. Confusion avec S. vespiformis Confusion avec plusieurs espèces. ATTRIBUT CD _ NOM NOM_COMPLET TYPE VALEUR Référentiel Code du taxon dans le référentiel TAXREF CharactèreString - SEMANTIQUE Code du taxon dans TAXREF Nom complet du taxon dans TAXREF Les attributs concernant la détermination et la validation des données sont les même pour les deux sexes Les attributs concernant la détermination et la validation des données sont spécifiques aux mâles Les attributs concernant la détermination et la validation des données sont spécifiques aux femelles Pas d'espèces proches, simple aspect visuel suffisant. Inutile de comparer avec d'autres espèces. Photo suffisante quelque soit l'angle ou la qualité. MIXTE Liste SEXE MALE FEMELLE TRES_SIMPLE Peu d'espèces ressemblantes, aspect visuel suffisant pour distinguer des espèces proches moyennant connaissance d'un critère, sans besoin de loupe. Détermination possible sur photo à partir d'une photo générale du spécimen. Simple sous conditions biogéographiques. Détermination délicate ou expert en raison de plusieurs espèces proches mais uniquement dans certains contextes biogéographiques. La détermination est simple dans les autres contextes car l'espèce est la seule présente. Simple sous conditions phénologiques. Détermination délicate ou expert en raison de plusieurs espèces proches mais la date d'observation est un critère reconnu pour la détermination. Plusieurs espèces ressemblantes (dans le même groupe taxonomique mais aussi éventuellement des apparences proches dans d'autres tribus ou sousfamilles). Besoin d'une comparaison avec d'autres spécimens ou d'une bonne habitude de la clé de détermination. Possible sur photo pour quelqu'un d'expérimenté, si le ou les critères sont mis en avant. Plusieurs espèces proches. Nécessité d'évaluer des critères subtiles en comparant les spécimens ou par dissection. Détermination sur photo impossible sauf dans le cas où un gros plan visualisant les critères est possible. Les espèces déterminables par séquençage du matériel génétique rentrent dans cette catégorie. Liste COL_IND COL_PG SEQ SANS FILET OBS_PG_TER OBS_IND_LAB EX_CRITERE_DET Liste OBS_PG_LAB PHOTO COL_IND COL_PG SEQ Une photo d'un individu, attachée à la donnée d'occurrence est considéré comme preuve associée à la détermination. Un exemplaire en collection, attaché à la donnée d'occurrence est considéré comme preuve associée à la détermination. La présence en collection des pièces génitales, attachées à la donnée d'occurrence est considéré comme preuve associée à la détermination. Le séquençage génétique (mitochondrial et/ou nucléaire), attachées à la donnée d'occurrence est considéré comme preuve associée à la détermination. Pas de critère de détermination exigé pour valider une donnée d'occurrence La détermination a été réalisée sur le terrain après examen de critères morphologiques spécifiques puis relâche de l'individu. Ce critère est exigé pour la validation des données d'occurrence. La détermination a été réalisée sur le terrain après examen des pièces génitales. Ce critère est exigé pour la validation des données d'occurrence. La détermination a été réalisée en laboratoire après comparaison d'un individu avec une collection de référence, sans garder cet individu en collection. Ce critère est exigé pour la validation des données d'occurrence. La détermination a été réalisée en laboratoire après comparaison des pièces génitales avec une collection de référence, sans garder les pièces génitales en collection. Ce critère est exigé pour la validation des données d'occurrence. La détermination est associée à une preuve photographique. Ce critère est exigé pour la validation des données d'occurrence. La détermination est associée à un individu en collection. Ce critère est exigé pour la validation des données d occurrence. La détermination est associée à des pièces génitales en collection. Ce critère est exigé pour la validation des données d'occurrence. La détermination est associée à un séquençage génétique. Ce critère est exigé pour la validation des données d'occurrence..	34,634
hal-01559568-978-3-642-18333-1_43_Chapter.txt_1	French-Science-Pile	Open Science	Various open science	2,017	Construction and Practice of Information Demonstration Area in Mentougou District of Beijing. 4th Conference on Computer and Computing Technologies in Agriculture (CCTA), Oct 2010, Nanchang, China. pp.367-373, ⟨10.1007/978-3-642-18333-1_43⟩. ⟨hal-01559568⟩	None	English	Spoken	2,089	3,145	Construction and Practice of Information Demonstration Area in Mentougou District of Beijing Juan Pan, Na Zhang, Shan Yao, Jian Xu To cite this version: Juan Pan, Na Zhang, Shan Yao, Jian Xu. Construction and Practice of Information Demonstration Area in Mentougou District of Beijing. 4th Conference on Computer and Computing Technologies in Agriculture (CCTA), Oct 2010, Nanchang, China. pp.367-373, �10.1007/978-3-642-18333-1_43�. �hal-01559568� HAL Id: hal-01559568 https://inria.hal.science/hal-01559568 Submitted on 10 Jul 2017 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 Construction and Practice of Information Demonstration Area in Mentougou District of Beijing Juan Pan, Na Zhang*, Shan Yao, Jian Xu Department of Computer and Information Engineering, Beijing University of Agriculture, Beijing, PR. China 102206 juan_pan@163.com Abstract. The rural informatization is one of the important foundation for the construction of the metropolis-modern agriculture, which Beijing government makes great effort to develop now. Based on current situation of rural informatization construction in Mentougou district of Beijing, this study established an information demonstration area in order to integrate the information from the local natural ecology, agricultural production, special products trading and government. We made use of the technology of 3S, database and network to achieve the digitalization and visualization of the rural information. The study helps to guide the agricultural production and agricultural products circulation and offers the effective decision support for the sustainable development of the demonstration area. Keywords: rural informatization, information demonstration area, 3S, information service platform, Eco-Agriculture. 1 Introduction With the balance development of modern rural and urban areas, Beijing strengthens the rural informatization construction. The rural information infrastructure in Mentougou district has made remarkable progress after years of effort. An integrated basic information service network has been established, which provides a strong guarantee for Mentougou informatization construction (Shi 2009). 3S and Internet technology provide a new mode for the management of rural information, which realizes the management of spatial information that can’t be carried out in the traditional one. The Commission of Science and Technology of Mentougou district has established three websites successively—High-quality Goods Website, Chinese Walnut Website and Ecology Commercial City Website. However, the first two websites do not have the background system, and the third one needs to improve its trading function. Furthermore, these three websites provide some redundant function, and their network systems are instable. According to the current situation of informatization construction in Mentougou district, the study established information demonstration area in order to strengthen coordination and integration of rural information resources, and effectively promote the development of information technology in rural areas. We made use of 3S, database and network technology to manage and develop all kinds of rural information. Through the digitalization and visualization of the rural information, the study helps to guide the agricultural production and agricultural products circulation and offers the effective decision support for the sustainable development of the demonstration area. 2 The Overview of Study Area Mentougou district is located in the southwest of Beijing, 62 kilometers east-west, 34 kilometers north-south, with a total area of 1,455 square kilometers. It is located at 115 °25′ 00″ ~ 116 °10′ 07″E, 39 °48′ 34″ ~ 40 ° 10′ 37″N. The mountain area accounts for about 98.5% of the whole area, and the plain accounts for 1.5%. The study area belongs to the middle latitude continental monsoon climate. It’s droughty and windy in spring, hot and rainy in summer, cool and moist in fall, cold and dry in winter. There is great difference between western mountainous area and eastern plain in climate, the annual mean temperature is 11.7℃ in the east and 10.2℃ in the west. There are three rivers flowing through the study area— the Yongding River, Daqing River and Beiyun River. Yongding River covers the largest drainage area, which is about 1,368.03 square kilometers. Mentougou integrated ecosystem is composed of mountains, green fields and water system, which is an important ecological barrier and water source conservation area for Beijing. Therefore Mentougou is the key district to ensure the sustainable development of the Capital. 3 Structure and Function of the System According to the requirement of the eco-conservation and terrain features, the study established the information demonstration area in order to improve the competitiveness of agricultural products, optimize and upgrade the rural economic structure, and transform the pattern of economic growth. The construction of information demonstration area made full use of the advantages of GIS in data management, information visualization and spatial information analysis, integrated the information of natural ecology, agricultural production, special products trading and government, and established an eco-agriculture information service platform. Two towns, Wangping and Miaofenshan, are firstly selected as the experimental units. Then the practice is extended to the whole district. 3.1 Natural Ecological Information Module This module provides information display and query of meteorological, natural vegetation, hydrology, geology, soil type and so on. The meteorological information mainly involves temperature, relative humidity, precipitation and sunshine hours. 3.2 Agricultural Production Information Module This module mainly records the information of agricultural resources to realize the network management of agricultural production process. The information includes farm fields, soil nutrient, soil fertility, varieties of agricultural products, planting area, tree age and its spatial distribution. The soil nutrient includes the organic matter, total nitrogen, available nitrogen, available phosphorus, available potassium and PH value. In addition, according to standardized production practice, this module records the information of water and fertilizer management , training and pruning , pest control, growth process of crop and so on. All of the above mentioned information can be displayed and queried with the field parcel as a unit. 3.3 Special Products Trading Module Mentougou district produces abundantly special products, such as pear, walnut, cherry, almond and apricot. Some products, named as the tributes to the imperial palace, are famous over China, especially the roses in Xiangjiangou Village, Miaofengshan town enjoy sound reputation both in domestic and overseas due to big flowers, thick petal, dense color, fragrant gout and high oil content. Special products trading module realizes online trading and product tracing. Online trading mainly includes product information publication and recommendation, agricultural product credit guarantees, payment function, distribution process and information flow records. For the function of tracing back the product information, the product identification codes are affixed to the smallest package. With the unique identification code, the platform can query the detailed records of production management, products processing, logistics distribution information, and builds the quality assurance system from the origin of product to the market. 3.4 Government Affair Management Module This module issues the related policies and regulations information, and manages the daily business of towns and villages. The module emphasizes on the statistics analysis of social and economic data to provide the basic economic evaluation. The data includes numbers of households, total population, per capital annual net income, per capital disposable income, annual wages income, household business income, annual property income, annual transfer income, total expenditure and others. 4 Implementing Scheme As shown in Fig.1, implementing scheme of the system is composed of data acquisition and processing, database construction and network platform construction. data acquisition remote sensing data GPS fundamental geographic data field survey data statistical data database construction platform frame functional division of the platform agricultural production information module natural ecological information module special products trading government affair management module Mentougou Eco-Agriculture information service platform Fig. 1. Implementing scheme of the system 4.1 Data Acquisition and Processing The basic data and maps need to be collected and integrated. The data includes satellite image map, basic map and thematic map, statistical data on rural economy over years, and other agricultural data of meteorology and envirnment and etc. 4.2 Database Construction Rural information resources mainly include natural resource information, ecological information, agricultural production management information, agricultural market information and government affair information. All the information are processed, formatted and saved in the information resource database, which can be easily saved, retrieved, transmitted, published and shared through modern information technology. According to the local conditions, the information demonstration area constructs the spatial and attribute database, among which spatial information service is the basic database. The accuracy, capacity, coverage and update of spatial database not only affect the current construction of information demonstration area, but also have farreaching influence on the economy of the district and the scientific value of the study. Spatial database integrates the information of topographic, traffic network, administrative division, soil type, soil nutrient, land use status, field parcel distribution , temperature，rainfall and so on. Attribute database includes agricultural production information, product trading information, product tracing and management of administrative villages. All the information is connected with field parcel data. 4.3 Network Platform Construction The study establishes Mentougou Eco-Agriculture information service platform, using .NET, SQL Server and SuperMap as development tools, C# as development language. The platform integrates four above-mentioned information management modules to realize the information query, analysis, decision-making, trading, trace back and others. The browser/server structure is applied to the system, and the client users can access the platform using a web browser. 5 Function Actualization of System Fig. 2. Visualization showing interface of Mentougou geographical space data This paper takes management of space data and visualization showing for example to show the function actualization of system. Visualization showing interface of Mentougou geographical space data is showed as Fig.2, which realizes management and visualization of data such as village boundary, road, water system, soil, vegetation, landform, farmland and soil using style, etc. The system can brings plane map as well as three-dimensional relief map, both of the ways of showing can realize the zooming in and zooming out of the map. In addition, relief map has flight function, which means the user can get access to related information of three-dimensional landform by user-defined route. The system provides two query methods: query map by attribute and query attribute by map. As shown in Fig.3, based on the year, names of town and administrative village selected by the user, the system shows relevant positional information. When the user clicks on the map, the system will show information from the attribute database, such as area, numbers of households, agricultural population, income, climate, fertility of soil, etc. Fig. 3. Query result of administrative village 6 Conclusion and Prospect The study establishes the information demonstration area based on the technology of .NET, 3S and database to realize the integration of rural information resources. Through fully utilizing the advantages of GIS in map expression, spatial data management and map processing, the information service platform improves the visualization and convenient of management of rural information, advances the management efficiency of rural information. Further study and discussion: (1) To provide technical support for the planting structure adjustment, product variety update and agricultural division based on further study on the database of climate and agricultural resources. (2) To establish the fertilizer recommendation system for green food according to the soil nutrient database of special products, and to provide scientific support for cultivation of high yield and quality with the study of the relation between soil nutrition status and product yield and quantity. (3) To establish the remote sensing monitoring system to monitor diseases and insect pests of special products and provide real reference for decision-making. (4) To build comprehensive evaluation model for ecological capacity maximizing the ecological, economic and social benefits in order to promote the virtuous circle of ecosystem, and provide decision support for sustainable development of the area. Acknowledgments This study was funded by the Commission of Science and Technology of Mentougou district, and the project number is D0804090041000. References He L.Y., Huang W., Guo Z.H., Miao J.: Status, Task and Problem of Information Demonstration Village Construction in China (in Chinese). In: ccta2007, pp. 409—415. (2007) Li M.: Demand Analysis and Development Strategy for Informatization of Rural Area in Beijing city (in Chinese). J. Agriculture Network Information. 47--49(2009) Shi Y.Q.: Reflections and Suggestions On Rural Informatization Construction in the Mountain Areas of Beijing (in Chinese). J. Agriculture Network Information. 41--44 (2009) Zhu H.J., Wu H.R., Feng C., Zhong X., Sun X.: The Application of GIS in the Information Service Platform for New Village Construction (in Chinese). J. Journal of Agricultural Mechanization Research. 164--166(2008).	38,821
https://openalex.org/W2514092235	OpenAlex	Open Science	CC-By	2,016	Emerging opportunities and challenges in phenology: a review	Jianwu Tang	English	Spoken	12,675	24,080	SPECIAL FEATURE: INTERNATIONAL LTER Emerging opportunities and challenges in phenology: a review Jianwu Tang,1,† Christian Körner,2 Hiroyuki Muraoka,3 Shilong Piao,4,5 Miaogen Shen,5 Stephen J. Thackeray,6 and Xi Yang7 Key words: cameras; greenness; ILTER; modeling; phenology; scale; Special Feature: International LTER. Key words: cameras; greenness; ILTER; modeling; phenology; scale; Special Feature: International LTER. Received 3 November 2015; revised 28 April 2016; accepted 26 May 2016. Corresponding Editor: K. Vanderbilt. Copyright: © 2016 Tang et al. 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. † E-mail: jtang@mbl.edu † E-mail: jtang@mbl.edu † E-mail: jtang@mbl.edu v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 Emerging opportunities and challenges in phenology: a review Jianwu Tang,1,† Christian Körner,2 Hiroyuki Muraoka,3 Shilong Piao,4,5 Miaogen Shen,5 Stephen J. Thackeray,6 and Xi Yang7 1Ecosystems Center, Marine Biological Laboratory, Woods Hole, Massachusetts 02543 USA 2Institute of Botany, University of Basel, Schönbeinstrasse 6, 4056 Basel, Switzerland 3River Basin Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1193 Japan 4Department of Ecology, College of Urban and Environmental Sciences, Peking University, Beijing 100871 China 5Key Laboratory of Alpine Ecology and Biodiversity, Institute of Tibetan Plateau Research, CAS Center for Excellence in Tibetan Plateau Earth Sciences, Chinese Academy of Sciences, 16 Lincui Road, Chaoyang District, Beijing, China 6Lake Ecosystems Group, Centre for Ecology & Hydrology, Lancaster Environment Centre, Bailrigg, Lancaster LA1 4AP UK 7Department of Earth, Environmental and Planetary Sciences, Brown University, Providence, Rhode Island 02912 USA Citation: Tang, J., C. Körner, H. Muraoka, S. Piao, M. Shen, S. J. Thackeray, and X. Yang. 2016. Emerging opportunities and challenges in phenology: a review. Ecosphere 7(8):e01436. 10.1002/ecs2.1436 Abstract. Plant phenology research has gained increasing attention because of the sensitivity of phenol- ogy to climate change and its consequences for ecosystem function. Recent technological development has made it possible to gather invaluable data at a variety of spatial and ecological scales. Despite our ability to observe phenological change at multiple scales, the mechanistic basis of phenology is still not well understood. Integration of multiple disciplines, including ecology, evolutionary biology, climate science, and remote sensing, with long-term monitoring data across multiple spatial scales is needed to advance understanding of phenology. We review the mechanisms and major drivers of plant phenology, including temperature, photoperiod, and winter chilling, as well as other factors such as competition, resource lim- itation, and genetics. Shifts in plant phenology have significant consequences on ecosystem productivity, carbon cycling, competition, food webs, and other ecosystem functions and services. We summarize recent advances in observation techniques across multiple spatial scales, including digital repeat photography, other complementary optical measurements, and solar-induced fluorescence, to assess our capability to address the importance of these scale-dependent drivers. Then, we review phenology models as an im- portant component of earth system modeling. We find that the lack of species-level knowledge and ob- servation data leads to difficulties in the development of vegetation phenology models at ecosystem or community scales. Finally, we recommend further research to advance understanding of the mechanisms governing phenology and the standardization of phenology observation methods across networks. With the opportunity for “big data” collection for plant phenology, we envision a breakthrough in process- based phenology modeling. Special Feature: International LTER Tang et al. Fig. 1. Trends of papers and citations of vegetation phenology each year between year 1970 and 2014. The statistics are from an ISI Web of Knowledge search conducted on 26 October 2015 using the following terms: (Title = Phenolog) AND (Topic = vegetation OR Topic = plant). Fig. 1. Trends of papers and citations of vegetation phenology each year between year 1970 and 2014. The statistics are from an ISI Web of Knowledge search conducted on 26 October 2015 using the following terms: Fig. 1. Trends of papers and citations of vegetation phenology each year between year 1970 and 2014. The statistics are from an ISI Web of Knowledge search conducted on 26 October 2015 using the following terms: (Title = Phenolog) AND (Topic = vegetation OR Topic = plant). phenological shifts are a ubiquitous phenomenon and can be interpreted as a sensitive indicator of the ecological impacts of climate change (Walther et al. 2002); (2) the consequences of phenological shifts, including the start, senescence, and the duration of growing seasons, significantly impact ecosystem structure and function, such as leaf area, photosynthesis, carbon cycling, and species composition and competition, but the mechanistic understanding of these links and feedbacks to cli- mate change is currently incomplete (Richardson et al. 2012); and (3) technological developments, particularly in remote sensing and near-surface observation, have radically expanded the scale of phenological observation beyond traditional man- ual observation (Zhang et al. 2003, Richardson et al. 2009, Yang et al. 2012, 2014). Despite these advances, a deeper mechanistic understanding of phenology, its variability and drivers across multiple scales, and its link to other physiological processes is still needed. The lack of this funda- mental understanding has limited our ability to develop predictive models (Basler 2016). to bring to light—referring to visible changes in biological development, modern phenological research should go beyond visible observation of biological events by looking deeply into the phys- iological mechanisms of these changes. Plants change the timing of leaf-out, leaf senescence, and flowering according to environmental cues, for example, temperature (Richardson et al. 2006, Vitasse et al. 2011), irradiance (Saleska et al. 2007), precipitation (Craine et al. 2012, Shen et al. 2015), and photoperiod (Körner and Basler 2010, Basler and Körner 2012). These cues or drivers may vary in importance at different scales, depending on species, biomes, and environmental constraints. Introduction decades. Since 1980, the number of peer-reviewed journal articles related to plant phenology has increased by about 10-fold (Fig. 1). The rejuvena- tion of this long-studied subdiscipline of ecology has been primarily induced by three factors: (1) Plant phenology, the study of recurring events in the life cycle of plants, has gained increasing public and scientific attention over the last few v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 1 Special Feature: International LTER v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 Tang et al. Tang et al. The study of phenology spans multiple spatial and ecological scales, each with its own suite of observational methods. While genetic tools have been used to understand the controls over flow- ering in model plants such as Arabidopsis thaliana (Chew et al. 2012, Satake et al. 2013), a search of the current literature (using the ISI Web of Knowledge) suggests that most studies focus on broader ecosystem scales, with remote sens- ing being one of the most frequently used tools. Satellite remote sensing provides data on total vegetative growth (usually using indices like the normalized difference vegetation index [NDVI]) on a daily or weekly basis over large areas (e.g., Fisher and Mustard 2007, Xu et al. 2013). Recently, the development of near-surface remote sensing techniques, such as digital repeat photography or spectroscopic sensors, has provided high- temporal-resolution (~hourly) data of vegetation activities. On the other hand, long-term ground observations, especially of some dozen-year-old or hundred-year-old historical data (Miller-Rushing et al. 2006), are still valuable and complementary to newer technologies as the former provides phenological data of individual plants and spe- cies often over a longer time period than those of available satellite or repeat photography data. The above three methods—in situ ground observa- tions, near-surface, and remote sensing—continue to provide rich data from multiple spatial scales and also opportunities to transfer mechanistic understanding from detailed local studies to the development of quantitative models. In this review, we briefly revisit the mech- anisms and drivers of plant phenology as a foundation for guiding future technological advancement and model development. Then, we summarize recent advances in observation tech- niques across multiple spatial scales using exam- ples drawn from ILTER sites. Finally, we review phenology models as an important component in earth system modeling. We close with rec- ommendations for future research directions in phenology, emphasizing the importance of long- term, multidisciplinary research. Special Feature: International LTER Therefore, lessons learned or models built from one species or ecosystem may not be widely applicable unless we possess the mechanistic insight to permit generalizations for other sys- tems. The variation in these responses to a chang- ing climate could be traced to genetic controls and evolutionary processes in addition to the environ- mental drivers (Wilczek et al. 2009). Phenology could also exert feedbacks to the climate system through changing leaf structure, energy balance, and carbon and water cycles (Penuelas et al. 2009). An integrated framework is urgently needed to advance our understanding of the mechanisms driving phenological shifts and their consequence on ecosystem structure, function, and biogeo- chemical cycles (Wolkovich et al. 2014). Integration of multiple disciplines, includ- ing ecology, evolutionary biology, and climate science, with remote sensing and long-term, high-frequency monitoring data across multiple spatial scales is essential to advance phenological research. Although the word “phenology” origi- nates from Greek φαίνω (phainō)—to show and v www.esajournals.org 2 August 2016 v Volume 7(8) v Article e01436 Mechanisms and Drivers of Plant Phenology Plant phenology responds sensitively to the changing environment for two general reasons: (1) to achieve synchrony in sexual reproduction among individuals of a given population (gene flow) and (2) to escape unfavorable seasons in a precautionary way. While the first can use any environmental signal, all individuals of a popula- tion have evolved as a result of the second reason, which requires signals to avoid exposure of unhardened or fragile tissues to damaging environmental conditions such as freezing tem- peratures. As concurrent weather is an intrinsi- cally unreliable predictor of extreme events, plant evolution in temperate climates has selected for the perception of signals that bear, in a probabilistic manner, a minimum likelihood of fatal exposure to extremes. Although not fully understood, it is clear that most species in temperate climates adopt a mix of signals that fall into three categories: (1) solar signals (photoperiod), (2) past seasonal experience signals (winter chilling), and (3) current or very recent past signals (concurrent tempera- ture and/or water conditions). When photoperiod is employed for spring phenology, winter chilling must be employed as well, because day length is the same in spring and autumn and thus the leg- acy of winter experience (chilling exposure) is needed to sensitize plants to spring or autumn (Schwartz and Hanes 2010, Polgar and Primack 2011). Alternatively, plants may solely rely on thermal forcing, as winter transitions to summer. However, the temperature signal is less reliable because exceptionally warm early spring weather may induce leaf-out before the statistical risk of freezing is over (i.e., a “false spring”; Allstadt The scope of phenology studies across time and space requires long-term research and interna- tional collaboration. For example, recording the interannual variation in leaf-out dates is essential to understanding the environmental cues of leaf- out. Without years of leaf-out data across species, ecosystems, and biomes, we will not be able to gen- eralize the responses of spring phenology to envi- ronmental change and, therefore, to predict the impact of future climate change. The International Long Term Ecological Research network (ILTER), a global network of approximately 600 research sites located in diverse ecosystems, plays a unique role in providing existing long-term phenology data across biomes and regions. The ILTER could also serve as a powerful mechanism for the standardiza- tion of future phenology data collection using new technologies to advance phenological research. August 2016 v Volume 7(8) v Article e01436 v www.esajournals.org Special Feature: International LTER Tang et al. climatic condition by minimizing competition. The genetic control of phenology limits the plas- ticity of phenological shifts and protects a species or population from climate extremes. For exam- ple, when the southern and northern populations of Eriophorum vaginatum, a dominant species in the arctic tundra, were transplanted to a central location in the Toolik Station in Alaska, an ILTER site, the dates of senescence of each population remained unchanged (i.e., northern population senesced earlier than the southern one despite the shift of temperature and photoperiod), indicating a genetic control of phenology for each popula- tion (T. Parker, J. Tang, M. Moody, and N. Fetcher, personal communication). Therefore, to fully under- stand phenology and its response to the changing climate, abiotic and biotic drivers should be dif- ferentiated and integrated into modeling efforts (Steltzer and Post 2009, Schwartz and Hanes 2010, Pau et al. 2011, Wolkovich et al. 2014). Insights from these studies can be used to guide the tech- nological development of phenological observa- tion (e.g., using high-resolution near-surface and remote sensing data to differentiate phenolog- ical differences across species that may respond differently to environmental drivers) and the experimental design (e.g., using warming and transplanting experiments to detect abiotic and biotic drivers of phenology). et al. 2015) or rainy season begins (Shen et al. 2015). Whatever combination of signals plants uti- lize, the influence of concurrent temperature will force development, once the internal requirement has been met (Körner and Basler 2010).f Different life history strategies among plants will select for less or more precautionary phe- nology. In spring, short-lived, highly reproduc- tive, so-called r-strategists will more likely adopt an opportunistic phenology (in the extreme case just following concurrent temperature), whereas long-lived, slowly reproducing plants (K-strategists) such as late-successional forest trees will adopt a strategy that ensures century- long life (Pianka 1970, Steltzer and Post 2009, Körner and Basler 2010, Wolkovich et al. 2014). Opportunistic plants may lengthen their growing season in warming springs, but late-successional, low-risk plants may take less advantage from a warmer spring. Hence, there is a trade-off between freezing tolerance and required season length that defines the distributional range of species that adopt different phenological strat- egies (Lenz et al. 2014, Vitasse et al. 2014a, b, Körner et al. 2016). Special Feature: International LTER In autumn, the late season transition to dor- mancy should occur before the advent of peri- ods that bear a risk of freezing damage. Most temperate and boreal trees use photoperiod as a reliable signal for the onset of autumnal hard- ening. Yet, unlike spring phenology that can be visually observed clearly (e.g., budbreak and leaf flushing), autumnal phenology cannot be read- ily assessed by visual signals. Leaf coloration in autumn is only a final step of a cascade of invis- ible developmental processes (such as winter bud set or leaf abscission layer formation), most commonly induced by unpredictable cold nights. Hence, satellite images, automated systems, or phenology modeling is intrinsically limited to assess “true” autumn phenology (Yang et al. 2012), because data on canopy color change do not tightly relate to the underlying processes of phenology in autumn (Körner et al. 2016). v www.esajournals.org Mechanisms and Drivers of Plant Phenology v www.esajournals.org 3 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER August 2016 v Volume 7(8) v Article e01436 Advancement of Measurement Methods and Long-term Data To advance understanding of the above pheno- logical mechanisms, drivers, strategies, and their variations among species will require extensive, intensive, and long-term measurements of phe- nological data. Multiscale, long-term phenologi- cal monitoring allows interpretation of the mechanisms behind observed phenology. The metrics for plant phenology measurements include budbreak, leaf expansion and matura- tion, flowering time, senescence (coloring), and leaf abscission. The methodology for such obser- vations includes the recording of phenological events by eyes, periodic photography, automatic repeat photography, and satellite-based remote sensing (Morisette et al. 2009). During the past decades, the leaf and canopy phenology of vege- tation has been focused on remote sensing obser- vations from a single research plot to regional, continental, and global scales to detect any phe- nological changes due to interannual variation in climatic conditions or ongoing global warming (Cleland et al. 2007, Richardson et al. 2007, Piao et al. 2008, 2015). During the last decade, camera- based observations have been popular with the advancement of digital repeat photography (Richardson et al. 2007, 2009, Ide and Oguma 2010, Nagai et al. 2011). In addition, spectroradi- ometers have become powerful tools to monitor the spectral reflectance of both individual leaves and whole canopies (Asner and Martin 2008). Such data can be compared with the reflectance measured by satellite-based remote sensing (Muraoka et al. 2013b) and with laboratory assays of leaf traits (Yang et al. 2014) that provide physi- ological information otherwise invisible when solely recording reflectance of visible bands. Given these different phenological strategies, observations obtained from one of the above groups may not necessarily apply to other groups. Phenology patterns found in ornamen- tal trees, domestic trees, exotic taxa, and more generally ruderal/pioneer taxa may not be fully applicable to late-successional forest trees. This species variation in the strategy to respond to climate should be considered when designing monitoring strategies and also developing mod- els to capture generic responses to environmen- tal change. Species within a plant community will likely not respond to climate change using the same phenological strategies, which may have signif- icant consequences for resource (i.e., light and nutrients) competition, food webs, ecosystem productivity, carbon cycling, and other ecosystem functions and services. Special Feature: International LTER Tang et al. Tang et al. evolutionary heritage from central or central- west Asia, with a very continental climate and abrupt and reliable onset of spring (Janick 2005). Hence, it is common in the orchard industry that plants suffer from frost after leaf-out in response to the conditions typical of an unpredictable spring. (4) Ornamental species: ornamental plants that exhibit extreme forms of opportunis- tic phenology in response to temperature, because they originate from regions with hardly any freezing risk and almost behave like a bio- logical thermometer, advancing their phenology by accumulating degree-days (Defila and Clot 2001). Examples are horse chestnut (Aesculus hip- pocastanum) and lilac (Syringa vulgaris; Larcher 2006).f measuring shifts in phenology, can be a powerful combination for better understanding the mecha- nisms that cause changes in phenology and their ecological consequences. evolutionary heritage from central or central- west Asia, with a very continental climate and abrupt and reliable onset of spring (Janick 2005). Hence, it is common in the orchard industry that plants suffer from frost after leaf-out in response to the conditions typical of an unpredictable spring. (4) Ornamental species: ornamental plants that exhibit extreme forms of opportunis- tic phenology in response to temperature, because they originate from regions with hardly any freezing risk and almost behave like a bio- logical thermometer, advancing their phenology by accumulating degree-days (Defila and Clot 2001). Examples are horse chestnut (Aesculus hip- pocastanum) and lilac (Syringa vulgaris; Larcher 2006).f Phenological Strategies and Consequences The complexity of biodiversity and multiple controls of plant phenology result in highly vari- able responses of plant phenology to climate among species, communities, and ecosystems (Rollinson and Kaye 2012, Shen et al. 2014a). There are four broad phenological strategies with which plants respond to the environment. (1) Opportunistic species: r-selected species, as described above, for which leaf-out in spring is less precautionary as it follows direct thermal forcing. (2) Strict species: K-selected species as described above that employ stronger photoperi- odic controls than opportunists. (3) Exotic spe- cies: exotic plants that are not adapted to their “new” climates. For example, orchard trees planted in humid-temperate climates, including apricot (Prunus armeniaca), cherry (Prunus avium), and apple (Malus domestica), have an In addition to the above abiotic cues, biological factors, including competition, resource limita- tion, and genetics, also control phenology, partic- ularly during the period of a stable environment (Wolkovich et al. 2014). Coexistence of species in a community often demonstrates a similar pheno- logical pattern of each species under the similar v www.esajournals.org 4 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER August 2016 v Volume 7(8) v Article e01436 v www.esajournals.org Advancement of Measurement Methods and Long-term Data Within the plant commu- nity, responses of canopy trees and understory shrubs and herbs to climate change may affect competition of these functional groups for spring light and soil nutrients (Vitasse 2013), and thus the overall ecosystem production and carbon cycling. Cross-species variation in phenolog- ical responses to climate change can also cause mismatch and disrupt the synchrony of species interactions. For example, under global warm- ing during the 1975–2000 period, winter moth (Operophtera brumata) eggs hatched earlier than oak (Quercus robur) bud burst in the Netherlands, profoundly affecting population dynamics of the winter moth (Visser and Holleman 2001). Advanced observational technologies, cou- pled with long-term human-collected data In the following sections, we summarize the recent advancement of methodologies for long- term monitoring of leaf and canopy phenology at multiple spatial scales. The methods and techniques used by satellite-based remote sens- ing have been widely reviewed elsewhere (e.g., Zhang et al. 2003, Homolova et al. 2013). v www.esajournals.org 5 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Special Feature: International LTER Tang et al. Tang et al. Tang et al. (http://asiaflux.net/), JaLTER (Japan Long Term Ecological Research network; http://www.jalter. org/), CERN (Chinese Ecological Research Net work; http://www.cern.ac.cn), LTER (Long Term Ecological Research network of the United States; http://www.lternet.edu), and other ILTER (Inter national Long Term Ecological Research network; http://www.ilternet.edu) sites as an integrated ele- ment of measured variables. Many of these data are now accessible online. Networks such as these have an important role to play in providing com- parable phenological data across ecosystems within each network. Calibration of cameras to make them interoperable across the networks is strongly needed (Wingate et al. 2015) to facilitate internetwork data comparisons. the green chromatic coordinate (Gcc; Sonnentag et al. 2012). GEI and Gcc are calculated as: (1) GEI=(G−R)+(G−B)=2G−(R+B); (2) Gcc=G∕(R+G+B); (2) where G is the green digital number from image files, R is the red digital number, and B the blue digital number. g These indices are widely used as a proxy to describe phenological patterns across the sea- son, either in a natural environment or in warm- ing experiments to detect phenological shifts in response to experimental warming (e.g., Chung et al. 2013). However, recent findings indicate that the camera-based greenness indices may not reveal leaf physiology that ties closely with pho- tosynthesis (Yang et al. 2014, Liu et al. 2015). For example, Yang et al. (2014) found that the camera greenness peaked 20 days earlier than the peak of leaf chlorophyll and carotenoids that are a bet- ter proxy for photosynthesis, and the decline of the greenness after the spring peak detected from cameras may not reflect the real change of the LAI. Further development of greenness indices or corrected indices that fully use visible-band camera images or expand the visible bands to broader bands is needed to reveal seasonality of physiological traits of plants. Within these networks, digital cameras for repeat photography are mounted on observa- tion towers to monitor leaf, branch, canopy, and landscape phenology. The cameras can be set to take pictures downward from the tower, upward from the ground, horizontally, or with fisheye views (Fig. 2). Fisheye images have been used to take hemispherical canopy photographs for esti- mating the light environment of the forest under- story (Anderson 1964, Chazdon and Field 1987, Pearcy 1989) and forest canopy leaf area index (LAI; Jonckheere et al. 2004). With the automa- tion of repeat fisheye images, continuous LAI data at the canopy scale are available. Other optical measurements p Visible-band images obtained from digital cameras could be expanded to other complemen- tary optical measurements to reveal more physi- ological and functional traits of plants. For example, to estimate gross primary production, one needs to know both the growing season length, which can be monitored by camera-based phenology, and photosynthetic rates. The latter could be directly measured by a photosynthesis system (e.g., LI-6400; LI-COR, Lincoln, Nebraska, USA), or estimated by a proxy, such as the leaf chlorophyll content. Chlorophyll contents are correlated with photosynthetic capacity of leaves throughout the seasons (Noda et al. 2015) and could be detected by measuring absorption of certain wavelengths by extracts of leaf pigments (Porra et al. 1989, Gitelson et al. 2003). The “SPAD meter” (Konica Minolta, Osaka, Japan) has been used to monitor the leaf chlorophyll content. The SPAD meter contains two light-emitting diodes Tang et al. In addi- tion, species-level observation with cameras has also been reported (Nijland et al. 2014). August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Special Feature: International LTER Tang et al. Digital repeat photography grasslands, and paddy fields (Richardson et al. 2009, Motohka et al. 2010, Nasahara and Nagai 2015). Camera-based phenology measurements allow direct observation of terrestrial vegetation from diverse habitats across the world and pro- vide data that can be linked with measures of ecosystem structure and functions. These cameras are currently located in many long-term ecologi- cal and carbon flux sites such as the AmeriFlux (http://ameriflux.lbl.gov/), NEON (National Eco logical Observatory Network of the United States; http://www.neoninc.org/), EUROFLUX, AsiaFlux g p p g p y With the advancement of commercial digital cameras, time-lapse cameras have become a widely used tool to monitor temporal changes in plants and the landscape. To observe terrestrial vegetation and its structure and function, and to link to remote sensing data from satellites, sensor networks, including “PhenoCam” (http:// phenocam.sr.unh.edu/webcam/), “European Phe nology Network (EPN),” and “Phenological Eyes Network (PEN)” (http://www.pheno-eye.org/), were established in multiple sites across forests, Fig. 2. Examples of digital repeat photography at the Takayama site (a JaLTER site, part of the ILTER) by the PEN camera systems. From the top to the bottom: branch of Betula ermanii, a canopy tree; downward view of the canopy; upward view of the canopy; landscape surrounding the forest. From left to right: early spring before leaf emergence; leaf expansion in late spring–early summer; canopy maturation in mid-summer; leaf coloring in autumn; leaf fall in late autumn. Fig. 2. Examples of digital repeat photography at the Takayama site (a JaLTER site, part of the ILTER) by the PEN camera systems. From the top to the bottom: branch of Betula ermanii, a canopy tree; downward view of the canopy; upward view of the canopy; landscape surrounding the forest. From left to right: early spring before leaf emergence; leaf expansion in late spring–early summer; canopy maturation in mid-summer; leaf coloring in autumn; leaf fall in late autumn. v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 6 Spatial scales: leaf, plot, regional, and global p f, p , g , g Phenology observations over a range of spatial and temporal scales are required to detect plant and ecosystem responses to changing environ- mental conditions (e.g., Cleland et al. 2007, Chung et al. 2013, Richardson et al. 2013). Leaf- scale phenology may not match canopy-scale phenology as the light conditions and nutrient supplies of leaves vary across the canopy height (Hirose 2005). Moreover, scaling canopy phenol- ogy to the region and the globe is challenging because species composition plays an important role at such larger scales and different species may respond differently to changing environ- mental conditions as summarized in earlier sections. Image processing Images taken by digital cameras contain values of red (R), green (G), and blue (B) as “spectral band” signals to detect the characteristics of the observed objects. For example, Richardson et al. (2007) and Nagai et al. (2011) examined the RGB signals as an indicator of phenological status of deciduous broadleaf forest canopies, and Saitoh et al. (2012) collected such data for an evergreen coniferous forest canopy, both confirming the capacity of this method to quantify canopy struc- ture (LAI) and function (photosynthetic capacity, leaf chlorophyll content, or eddy-covariance- based gross primary production). The RGB data are converted to greenness indi- ces to quantify the seasonal change, either as the green excess index (GEI; Richardson et al. 2007, Saitoh et al. 2012, Nagai et al. 2014) or as v www.esajournals.org 7 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Special Feature: International LTER Tang et al. be proportional to photosynthetic rates, and thus act as a proxy of photosynthesis at various scales. However, recording the fluorescence signal is extremely difficult because the weak radiation is obscured by the much higher intensity of solar radiation and its reflection from plants. Fortunately, because of the existence of a few absorption bands at which the atmosphere absorbs the solar radia- tion, for example, at the oxygen absorption wave- lengths at 687 and 760 nm, measuring the very weak signal of fluorescence is possible (Daumard et al. 2010, Guanter et al. 2013). With the newly launched satellites GOSAT (Greenhouse Gases Observing Satellite), GOME (Global Ozone Monitoring Experiment) 2, and OCO (Orbiting Carbon Observatory) 2, we are able to monitor flu- orescence across the globe, and potentially derive GPP and analyze the controlling factors and effects of stresses across space and time (Joiner et al. 2011, Guanter et al. 2012, Lee et al. 2013). More recently, ground-based SIF has been continuously moni- tored and found to have a strong correlation with GPP measured from an independent method (the eddy-covariance method) at Harvard Forest, an ILTER site (Yang et al. 2015), showing a great potential of using SIF to advance understanding of phenology and other ecosystem functions. for 650 and 940 nm, and a photodiode detector measures the transmittance of red and infrared light (Markwell et al. 1995). The SPAD values have to be validated against chlorophyll content data, extracted from the leaves to convert the val- ues to chlorophyll contents (Markwell et al. 1995). Simultaneous measurements of SPAD and leaf photosynthetic capacity are useful to investi- gate the phenology of the leaf physiological sta- tus over the season (Muraoka and Koizumi 2005, Liu et al. 2015). In addition, by combining with the RGB information obtained by a digital cam- era, SPAD values provide insights into the eco- physiological background of seasonal changes of canopy color (Nagai et al. 2011, Liu et al. 2015). for 650 and 940 nm, and a photodiode detector measures the transmittance of red and infrared light (Markwell et al. 1995). The SPAD values have to be validated against chlorophyll content data, extracted from the leaves to convert the val- ues to chlorophyll contents (Markwell et al. 1995). Special Feature: International LTER Simultaneous measurements of SPAD and leaf photosynthetic capacity are useful to investi- gate the phenology of the leaf physiological sta- tus over the season (Muraoka and Koizumi 2005, Liu et al. 2015). In addition, by combining with the RGB information obtained by a digital cam- era, SPAD values provide insights into the eco- physiological background of seasonal changes of canopy color (Nagai et al. 2011, Liu et al. 2015). To quantitatively measure the leaf expansion within the canopy over the season, optical proper- ties at the canopy scale, either reflectance or trans- mittance, could be recorded to capture the foliage and branch optical properties (Verhoef 1984). The seasonality of leaf expansion can be monitored by measuring the transmittance of photosyn- thetically active photon flux density above and below the canopy with light sensors to estimate LAI (Pearcy 1989, Hirose 2005, Monsi and Saeki 2005, Tang et al. 2008). In addition, the spectral reflectance of the canopy recorded by a spectro- radiometer that covers wavelengths beyond the visible bands of the solar spectrum can be used to estimate the structural and biochemical condi- tions of the canopy, for example, as the NDVI. The spectral reflectance data and vegetation indices obtained by satellite remote sensing (Piao et al. 2003, Motohka et al. 2010, Muraoka et al. 2013b) have been used to monitor physiological charac- teristics of vegetation such as pigment contents, leaf nitrogen, and water contents (Nakaji et al. 2007, 2008, Hilker et al. 2010, Asner et al. 2014, Stagakis et al. 2014, Yang et al. 2014). August 2016 v Volume 7(8) v Article e01436 Solar-induced fluorescence Solar-induced chlorophyll fluorescence (SIF) has emerged in recent years as a new tool to monitor beyond “visible” leaf color by focusing an “invisi- ble” ecosystem function, that is, photosynthesis (Joiner et al. 2011, 2013, Yang et al. 2015). SIF is a by-product of photosynthesis: When CO2 is assim- ilated under sunlight, a small fraction of solar energy (< 1%) captured by chlorophyll is emitted as fluorescence at a longer wavelength (650– 800 nm; Baker 2008, Meroni et al. 2009). SIF could Relationships between leaf ecophysiological characteristics and optical properties at smaller scales, from the single leaf, tree crown to forest canopy, have been used to estimate phenolog- ical changes at broader spatial scales from the landscape to regions, continents, and the globe (Muraoka and Koizumi 2009). This approach has v www.esajournals.org 8 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Tang et al. Tang et al. been introduced to ILTER-EAP (East Asia–Pacific regional network of ILTER) and J-BON (Japan Biodiversity Observation Network; Muraoka et al. 2013a). Vegetation indices obtained from satellite-based remote sensing data such as MODIS (Moderate Resolution Imaging Spectroradiometer) have been validated with ground-based data. The following vegetation indices have been introduced: been introduced to ILTER-EAP (East Asia–Pacific regional network of ILTER) and J-BON (Japan Biodiversity Observation Network; Muraoka et al. 2013a). Vegetation indices obtained from satellite-based remote sensing data such as MODIS (Moderate Resolution Imaging Spectroradiometer) have been validated with ground-based data. The following vegetation indices have been introduced: net ecosystem production (NEP) or net primary production (NPP) of ecosystems estimated by the eddy-covariance techniques (Xiao et al. 2004, Sims et al. 2006). But questions still remain as to whether the spectral information really expresses NEP or NPP as they are the final products of physiological processes including photosynthe- sis, respiration (both autotrophic and hetero- trophic), and biomass accumulation, which are reflected by the large difference in the phenology derived from EVI and canopy photosynthesis across the various vegetation types of AmeriFlux sites (Shen et al. 2014b). It remains a challenge to link the spectral information from the canopy and ecological processes involved within the ecosystems. (3) NDVI=(NIR−R)∕(NIR+R), (4) EVI=2.5(NIR−R)∕(NIR+6R−7.5B+1), (5) GRVI=(G−R)∕(G+R), (3) (5) where NDVI stands for the normalized differ- ence vegetation index, EVI for the enhanced veg- etation index (Huete et al. 2002), GRVI for the green-red vegetation index (Motohka et al. Solar-induced fluorescence 2010), NIR is the near-infrared band (841–876 nm), R is the red band (620–670 nm), B is the blue band (459–479 nm), and G is the green band (545– 565 nm) for MODIS.f where NDVI stands for the normalized differ- ence vegetation index, EVI for the enhanced veg- etation index (Huete et al. 2002), GRVI for the green-red vegetation index (Motohka et al. 2010), NIR is the near-infrared band (841–876 nm), R is the red band (620–670 nm), B is the blue band (459–479 nm), and G is the green band (545– 565 nm) for MODIS.f In summary, recent technological advances have made it possible to collect tremendous data sets on plant phenology at high temporal frequency with different spatial resolutions. Although pho- tographs and the spectral reflectance data have provided us with “visual” or “color” informa- tion that is useful for phenology observations, ecophysiological observation is also crucial to understanding the physiological mechanism underlying spectral information and the response of phenological events to the climatic conditions and their influence on photosynthetic productiv- ity of leaves, canopies, individual plants, and veg- etation (Vitasse et al. 2009, Muraoka et al. 2010, Chung et al. 2013, Yang et al. 2014). Normalized difference vegetation index has been found to match well with canopy LAI at low LAI (< 3 m2/m2; Potithep et al. 2013, Yang et al. 2014), while EVI matched well with the maximum GPP, calculated by a process-based model which incorporates the phenology of LAI and Vcmax (maximum velocity of carboxylation calculated from on-tower measurements of pho- tosynthesis/intercellular CO2 concentration rela- tionship by a portable photosynthesis system, LI-6400; LI-COR; Muraoka et al. 2013b). GRVI was found to be difficult to use to monitor the seasonal changes in these forest parameters, but Motohka et al. (2010) have found it to be useful to detect the date of onset (leaf expansion) and offset (leaf fall) in forests and grassland. v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Th l k f i l l k l d d b uptake (Piao et al. 2007, Keenan et al. 2014, Saitoh et al. 2015), while delayed autumn is found to lead to either increase (Keenan et al. 2014) or decrease in net carbon uptake (Piao et al. 2008). However, large uncertainties exist in current approaches to model spring and autumn phenology within ter- restrial ecosystem models (Keenan et al. 2012, Richardson et al. 2012), of which some are used as land surface models within earth system models in an attempt to forecast future climate. t p Both species-level and ecosystem-level mod- els are limited in predictive power without the understanding of the physiological mechanisms driving phenology. They may only reflect cor- relative findings within specific spatiotemporal scales. A model that relies on statistical correla- tions between phenology and past climates may not be able to project plant phenology into new combinations of chilling experience, photope- riod, and actual thermal forcing in the future. Although some plant phenology models are based (wholly or partly) on known or hypothet- ical cause–effect relationships between physi- ological processes and environmental factors (Chuine et al. 2003, Hanninen and Kramer 2007), most rest on statistical relationships obtained from past observations because of the lack of true dose–response functions. In general, these models consider one or two development phases before leaf unfolding or flowering, endodor- mancy and ecodormancy. Endodormancy is induced by a decreasing photoperiod and/or temperature in the autumn and is often broken after a period of “chilling” of low temperatures (Chuine et al. 2003, Horvath 2010), with the role of photoperiod commonly unknown or unac- counted for. The ecodormancy phase begins at the end of the endodormancy phase and finishes after a period of increasing temperature (Chuine 2000, Fu et al. 2012) in spring. Based on these cri- teria and responses, dozens of phenology mod- els (Chuine et al. 2003, Schaber and Badeck 2003, Fu et al. 2012, Olsson et al. 2013, Fila et al. 2014), most of them for tree species, have been devel- oped, commonly adopting the following gen- eral function (see Chuine et al. Special Feature: International LTER Tang et al. Tang et al. uptake (Piao et al. 2007, Keenan et al. 2014, Saitoh et al. 2015), while delayed autumn is found to lead to either increase (Keenan et al. 2014) or decrease in net carbon uptake (Piao et al. 2008). However, large uncertainties exist in current approaches to model spring and autumn phenology within ter- restrial ecosystem models (Keenan et al. 2012, Richardson et al. 2012), of which some are used as land surface models within earth system models in an attempt to forecast future climate. where tn and tn−1 are the dates at the end of devel- opment phases n and n − 1, respectively, Rn,t is the rate of development in phase n on day t, Z is the environmental factor such as temperature or photoperiod, Sn,t is the state of development in phase n on day t, and Sn is the critical state to finish the development phase (Chuine et al. 2003). One may take a chilling-forcing model for leaf unfolding, for example, with S1,t rep- resenting the sum of chilling (endodormancy phase) and S2,t being the accumulative degree- days (ecodormancy phase). To specify Eq. 6 for timing phenology, one needs to determine the number of phases, parameters and functional type of Rn,t(Z), and the critical value of Sn. For example, in a chilling-forcing model, one needs to determine the period, the functional type of development, base temperature, and the critical value for chilling sum (Chuine 2000). For a given species and in a given location, such a task is possible, with the predicted phenological timing differing from observations by a few days only (e.g., Jeong et al. 2013), if the observations and knowledge of physiological processes related to chilling and forcing are sufficient and if photope- riod does not matter. However, in the real world, such knowledge and observation are usually insufficient for most species and thus effective phenological models for these species are lack- ing. We are still not clear about the physiological basis of the endodormancy and the transition to ecodormancy related to chilling experience. In a recent study based on 490 species, only about 20% showed a relationship between phenology and winter temperature in regression analysis (Cook et al. 2012). We can only infer that for the other 80%, either chilling is not required, or the chilling requirement is not met by the measures of temperature employed for defining chilling. Phenology Modeling With knowledge gained from long-term moni- toring of phenological events and improved understanding of key drivers and mechanisms, we can build phenology models to simulate the timing of phenology and to project likely future change. Models can also be used to understand how climate-induced phenological changes may affect ecosystem functions such as carbon cycling and energy flows and their feedbacks to the cli- mate system (Richardson et al. 2013). In particular, changes in the timing of the onset and offset of the vegetation growing season have a considerable influence on the annual carbon budget in Northern Hemisphere vegetation (Piao et al. 2007, 2008, Richardson et al. 2010, Keenan et al. 2014). For example, a warming-induced advance in spring leaf emergence is suggested to increase net carbon f g Ground-based optical measurements have been used to upscale spectral vegetation indices and forest canopy photosynthesis to larger areas. Muraoka et al. (2013b) applied the relationship between in situ EVI and the maximum daily GPP to EVI data of MODIS/Terra for central Japan and found that the relationship between EVI and GPP showed seasonal hysteresis, suggest- ing that spectral reflectance and photosynthetic capacity of a forest canopy are not always one- to-one. A similar approach has been taken by combining such spectral vegetation indices and v www.esajournals.org 9 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER Tang et al. Tang et al. stages of species have their own strategies in response to environmental cues. It is difficult to aggregate all these responses into a single model to simulate vegetation phenology at ecosystem or regional scales. Even though we may use the same model structure for ecosystem-scale phe- nology as the species-scale (taking the chilling- forcing model for example), there are still several challenges, including selecting an aggregated response type, threshold temperatures above or below which forcing or chilling takes place, time periods for calculating the status of chilling and forcing, temperature data to be used (i.e., daily mean, maximum, minimum, or daytime mean temperature), and critical status of chilling and forcing. It is more complicated if we consider different responses to the same environmental factor among various species and multiple envi- ronmental factors and if the species composition of a community evolves with long-term climate change. Therefore, it is challenging to develop vegetation phenology models by integrating the phenology response of all the coexisting species within a community. However, recent advances 2012, Yang et al. 2012) at the ecosystem scale where vegetation types are derived from satellite data (Botta et al. 2000). Those vegetation phenology models are often driven by meteorological data to simulate seasonal leafing dynamics such as LAI (Richardson et al. 2012). Such simulations, how- ever, show large biases and only explain a limited fraction of interannual variation in observed phe- nological dates although some could perform bet- ter than others for a certain vegetation type, which further leads to poor prediction of GPP (Lafont et al. 2012, Richardson et al. 2012). Such a model would firstly suffer from the large uncertainties in the satellite-derived (statistical) phenology data (Botta et al. 2000, Picard et al. 2005, Cong et al. 2012), particularly for the nonforested vegetation at the middle and high latitude (Cao et al. 2015) and the tropical forest of which the phenology is difficult to capture by satellite data. 2012, Yang et al. 2012) at the ecosystem scale where vegetation types are derived from satellite data (Botta et al. 2000). Those vegetation phenology models are often driven by meteorological data to simulate seasonal leafing dynamics such as LAI (Richardson et al. 2012). Special Feature: International LTER 2003 for a list of species-level models): p p yi g g The lack of species-level knowledge and obser- vation data further leads to difficulty in develop- ing vegetation phenology models at ecosystem or community scales, particularly for nonwoody vegetation types, although those models are essen- tial in some terrestrial ecosystem models that are embedded in earth system models. Hence, most of the current phenology models have the same structure and similar functional type to the species- level ones to describe the effect of environmental drivers (Botta et al. 2000, Krinner et al. 2005, Knorr et al. 2010, Migliavacca et al. 2012, Richardson et al. tn suchthatSn,t = tn ∑ tn−1 Rn,t(Z)=S∗ n (6) v www.esajournals.org v www.esajournals.org v www.esajournals.org 10 August 2016 v Volume 7(8) v Article e01436 Special Feature: International LTER v www.esajournals.org Conclusions and Recommendations for Future Research With an ever-increasing array of techniques for observing phenology at a range of different scales, phenology research has entered a new stage that needs to integrate all relevant disciplines and take advantage of new technologies and long-term data collection. We emphasize that while photographs and spectral data are important to phenological research, acquiring better understanding of the physiological mechanisms governing phenology is essential. When mechanistic developments match advances of observation techniques, we will be able to not only understand drivers of pheno- logical change at multiple scales, but also greatly improve predictive modeling capabilities. Fig. 3 summarizes a framework of integrated phenologi- cal measurements, mechanisms, and modeling to advance our understanding of phenology and knowledge gaps (dashed lines) for further research. We recommend more studies to advance understanding of the mechanisms governing phenology and to disentangle the interacting effects of temperature, photoperiod, and win- ter chilling, as well as genetic controls and evo- lutionary dynamics for individual species. In addition to this essential ecophysiological under- standing, we should also address consequences of phenological shifts for ecological processes at multiple scales, their geographical variation such as along altitudinal and latitudinal gradients, the teleconnection of ecological functions (Heffernan et al. 2014), and the influence of, and response to, climate change. Further, standardization and interoperability of recent technologies in observ- ing phenology over long temporal scales, such as using digital cameras and spectroradiometers, from plot to regional scales are critically needed. International research networks such as ILTER can play an important role in standardizing phe- nology research protocols. Automatic repeat cameras that are able to conveniently and con- tinuously record phenology should be installed With an ever-increasing array of techniques for observing phenology at a range of different scales, phenology research has entered a new stage that needs to integrate all relevant disciplines and take advantage of new technologies and long-term data collection. We emphasize that while photographs and spectral data are important to phenological research, acquiring better understanding of the physiological mechanisms governing phenology is essential. When mechanistic developments match advances of observation techniques, we will be able to not only understand drivers of pheno- logical change at multiple scales, but also greatly improve predictive modeling capabilities. Fig. 3 summarizes a framework of integrated phenologi- cal measurements, mechanisms, and modeling to advance our understanding of phenology and knowledge gaps (dashed lines) for further research. Special Feature: International LTER Such simulations, how- ever, show large biases and only explain a limited fraction of interannual variation in observed phe- nological dates although some could perform bet- ter than others for a certain vegetation type, which further leads to poor prediction of GPP (Lafont et al. 2012, Richardson et al. 2012). Such a model would firstly suffer from the large uncertainties in the satellite-derived (statistical) phenology data (Botta et al. 2000, Picard et al. 2005, Cong et al. 2012), particularly for the nonforested vegetation at the middle and high latitude (Cao et al. 2015) and the tropical forest of which the phenology is difficult to capture by satellite data. Another challenge to developing ecosystem- scale phenology models emerges from the dif- ference in the phenology of different species that assemble a community. As we described earlier, different functional groups and successional Fig. 3. Integrating long-term, cross-scale phenology measurements and modeling to improve understanding of phenology, its response to climate change, and underlying physiological mechanisms. Arrowed lines indicate drivers and informational inputs; dashes indicate underdeveloped. Hierarchical drivers of temperature, chilling, and photoperiod indicate that (1) temperature alone, (2) temperature + chilling, or (3) temperature + photoperiod + chilling drive phenology. Fig. 3. Integrating long-term, cross-scale phenology measurements and modeling to improve understanding of phenology, its response to climate change, and underlying physiological mechanisms. Arrowed lines indicate drivers and informational inputs; dashes indicate underdeveloped. Hierarchical drivers of temperature, chilling, and photoperiod indicate that (1) temperature alone, (2) temperature + chilling, or (3) temperature + photoperiod + chilling drive phenology. v www.esajournals.org August 2016 v Volume 7(8) v Article e01436 11 Special Feature: International LTER Tang et al. Tang et al. of phenology. These cameras should be com- plemented by spectroradiometers to monitor leaf- and canopy-scale ecophysiology to capture additional physiological variables other than color information alone. The development of low-cost hyperspectral cameras along with new spectral indices and new devices to capture solar-induced fluorescence to track in situ vegetation activities is strongly recommended. As these observation sys- tems are connected as a network, they can provide phenological observations across multiple scales, from canopy, landscape, regional to the global. Such networks that also offer long-term human- collected data about phenological events are a rich basis for cross-scale phenological data synthesis. Conclusions and Recommendations for Future Research Given the opportunity of collecting unprec- edented phenology data with advanced tech- nologies, we envision a breakthrough in process-based phenology modeling, once biolog- ical drivers, dose–response functions, or thresh- old functions are available, rather than building solely upon correlative statistics based on past observations. The phenology models will need to account for the rate of microevolutionary adjust- ments of genotypic phenology controls. The cur- rent conspecific variability in phenology should not be simply considered “noise” but rather the starting point of evolutionary selection within subpopulations. Our understanding of individ- ual species’ phenological strategies in response to a changing environment remains incomplete and thus limits our ability to develop robust phe- nology models at the ecosystem scale. With these advancements, phenology models would be bet- ter parameterized and represented within earth system models. We recommend more studies to advance understanding of the mechanisms governing phenology and to disentangle the interacting effects of temperature, photoperiod, and win- ter chilling, as well as genetic controls and evo- lutionary dynamics for individual species. In addition to this essential ecophysiological under- standing, we should also address consequences of phenological shifts for ecological processes at multiple scales, their geographical variation such as along altitudinal and latitudinal gradients, the teleconnection of ecological functions (Heffernan et al. 2014), and the influence of, and response to, climate change. Further, standardization and interoperability of recent technologies in observ- ing phenology over long temporal scales, such as using digital cameras and spectroradiometers, from plot to regional scales are critically needed. Special Feature: International LTER in monitoring methods and cumulative phenol- ogy data from long-term research networks are paving the way for developing vegetation phe- nology models at large scales as a critical compo- nent of earth system models (Basler 2016). v www.esajournals.org Literature Cited Allstadt, A. J., S. J. Vavrus, P. J. Heglund, A. M. Pid- geon, W. E. Thogmartin, and V. C. Radeloff. 2015. Spring plant phenology and false springs in the conterminous US during the 21st century. Environ- mental Research Letters 10:104008. http://dx.doi. org/10.1088/1748-9326/10/10/104008. Cleland, E. E., I. Chuine, A. Menzel, H. A. Mooney, and M. D. Schwartz. 2007. Shifting plant phenology in response to global change. Trends in Ecology & Evolution 22:357–365. 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Photoperiod sensitiv- ity of bud burst in 14 temperate forest tree species. Agricultural and Forest Meteorology 165:73–81.t y Fila, G., M. Gardiman, P. Belvini, F. Meggio, and A. Pitacco. 2014. Acknowledgments JT was partially supported by the U.S. National Science Foundation grants PLR-1417763, DBI-959333, and AGS-1005663 and the University of Chicago and the MBL Lillie Research Innovation Award. HM thanks “PEN” developmental team and colleagues at the Takayama site and JaLTER, especially Nagai, Nasahara, Saitoh, Noda, Nakaji, and Hiura, and his research has been supported by NEXT Program and KAKENHI (MEXT, Japan). MS is partly supported by a fund from the National Science Foundation of China (grant 41571103). SJT was supported by an NERC grant NE/ J02080X/1. We thank comments and edits by two anon- ymous reviewers, Kristin Vanderbilt, Mary Heskel, and International research networks such as ILTER can play an important role in standardizing phe- nology research protocols. 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https://openalex.org/W1890714333_2	Spanish-Science-Pile	Open Science	Various open science	null	None	None	Spanish	Spoken	3,233	6,005	3.4. Petróleos Mexicanos (Pemex), Secretaría de la Defensa Nacional (Sedena), Secretaría de Marina (Secmar) De manera general, los servicios de salud prestados por estas instituciones registran una importante desconcentración en los dos últimos años contemplados en el análisis. A pesar de ello, estos servicios siguen beneficiando con mucha diferencia a la mitad más rica de la población. Cuando se analiza el tipo de atención médica por decil, se reproduce prácticamente el mismo patrón, con una pérdida importante para el 10% con mayores ingresos y mayor beneficio para el resto, especialmente los deciles medios-altos (sexto a noveno), puesto que 10% de las familias más pobres están literalmente ausentes en los servicios que prestan estas instituciones. Otra característica es que el perfil de los beneficiados es en su mayoría urbano, mientras que en el ámbito rural, prácticamente 50% de las familias con menores ingresos no recibe transferencia alguna por parte de estas instituciones. Resumiendo la estructura de los beneficios por parte de las instituciones proveedoras de salud, se puede decir que la ss, y en menor medida el imss, tienden a beneficiar a una población con menores recursos. Los cuidados preventivos son más importantes en la ss y los hospitalarios en el imss; sobre todo hacia la población urbana en el caso del imss y la población rural en el de la ss. Por su parte, instituciones como el issste, Pemex, la Sedena y la Secmar, atienden a un nicho específico de la población que les confiere una procuración de salud más orientada a familias de ingresos medios-altos y altos, en especial urbano y con mayor énfasis en servicios curativos y hospitalarios (cuadros 9-11). 12 Trabajadores al servicio del Estado. 748 Vargas-Téllez, C. O.: Progresividad del gasto público en salud... 4. Progresividad de las transferencias Existe una abundante literatura con igual gran cantidad de índices de desigualdad, algunos simplemente contienen datos estadísticos y otros también incluyen juicios de valor, de esta manera surge una importante disyuntiva cuando se trata de elegir un índice, la cual consiste en buscar un equilibrio entre atractivo estadístico y facilidad de interpretación con contenido normativo. Las propiedades de los índices son un buen indicador a la hora de elegir alguno, por ejemplo Shorroks (1983) apunta que el grado de sensibilidad a las transferencias es la característica más distintiva que debe tener un índice. Los índices elegidos para el presente análisis han de cumplir con el principio de transferencia débil, el cual se da cuando una transferencia a un individuo con ingreso (y), por parte de otro individuo más rico con ingreso y + δ, donde dicha transferencia es inferior a δ/2, hace que la desigualdad disminuya. El teorema sostiene: “Supóngase que la distribución del ingreso en el estado social A pueda ser alcanzado por una simple redistribución del ingreso en el estado social B (manteniéndole siempre constante el ingreso) y la curva de Lorenz para A se ubica totalmente dentro de la de B. Entonces, en la medida que se satisfaga el principio de transferencia débil, la medida de desigualdad siempre indicará un estrictamente menor nivel de desigualdad para el estado A que para el estado B” (Lambert, 2001: 55). Otro factor que se tomó en cuenta en la elección de los índices de concentración y de Kakwani para medir la progresividad de las transferencias en salud, es su fácil interpretación, ya que tienen como base el índice de Gini y la curva de Lorenz. Respecto a la interpretación, para los casos donde hay transferencias, los menores índices de concentración implican mejor distribución para los ingresos más bajos, lo que implica la misma lógica del coeficiente de Gini; de hecho, cuando se tornan negativos, indican que los recursos transferidos se concentran en las familias con menores ingresos y se dice que la transferencia en términos absolutos es totalmente redistributiva. Una vez explicadas brevemente la lógica y justificación de los índices a utilizar, se procede a su interpretación. Así, el coeficiente de concentración de las trasferencias totales en salud para los diferentes años de estudio, muestra una tendencia decreciente, lo que implica que este tipo de trasferencias sean en el último año de análisis casi neutrales, después de mostrar en un inicio cierta preferencia hacia los mayores ingresos. Sin embargo, por tipo de institución proveedora de salud, existen divergencias importantes (cuadro 12). Economía, Sociedad y Territorio, vol. xi, núm. 37, 2011, 729-757 749 Los servicios de salud adscritos a la Secretaría de Salud son los únicos que registran índices de concentración negativos para las cuatro encuestas analizadas, el comportamiento de las transferencias va desde una ligera progresividad cercana a la neutralidad en 1984, hasta una clara progresividad para el año 2002, con un coeficiente de concentración de -0.1136. El coeficiente de concentración de imss ha tenido un comportamiento errático con respecto al año de inicio del análisis, con una disminución en 1989 explicado ya por la posible inclusión del Programa imss-Coplamar, un aumento en 1996 y finalmente una ligera caída en el año 2002, de manera que ha permanecido casi invariable en 18 años, si acaso aumentó ligeramente la ventaja hacia los altos ingresos. Por su parte, el issste muestra los más altos coeficientes de concentración, con un incremento de 27% durante los años estudiados, lo que significa que los servicios de esta institución han tendido a concentrarse en las familias con mayores ingresos, empeorando el efecto redistributivo que pudiesen tener sus servicios. A manera de conclusión, se observan dos claros patrones en cuanto a progresividad para los servicios de salud: por un lado los servicios prestados por la Secretaría de Salud, que son los que benefician a la población abierta y muestran una tendencia hacia la progresividad; y por otro, los servicios prestados a la población asegurada por las dos principales instituciones públicas de seguridad social, los cuales muestran una claro patrón regresivo. Aunque cuando se considera el sistema completo, se advierte un ligero sesgo hacia la progresividad, conducido y soportado en su totalidad por los servicios universales prestados por la ss, los cuales benefician a la población abierta. Con base en el índice de Kakwani, el cual trata de cuantificar la desproporcionalidad de las transferencias en salud y, por tanto, la distancia entre la curva de Lorenz del ingreso disponible y de concentración, se pretende capturar su nivel de progresividad. Las transferencias en salud también se muestran bastante progresivas, incluso mayores que en educación, con altos valores en el índice de Kakwani y un aumento de 13.6% durante el periodo analizado. Sin embargo, cuando se observa la evolución por institución proveedora aparecen diferencias importantes. Mientras los servicios de la ss registran un índice de 0.621 y un aumento de 13% en su valor, los ofrecidos por el issste muestran un índice de apenas 0.036 y una caída en su valor de 72%; en tanto los ofrecidos por el imss se encuentran en valores intermedios. De esta manera existe un claro contraste entre la progresividad y evolución por tipo de institución, donde la alta progresividad de los servicios de la ss se ven contrarrestados por la nula progresividad del issste (cuadro 12). 750 Vargas-Téllez, C. O.: Progresividad del gasto público en salud... Cuadro 12 Transferencias en salud por tipo de institución Índices de concentración Tipo de transferencia Total ssa imss issste Tipo de Transferencia Total ssa imss issste 1984 1989 1996 2002 0.1096 0.0424 -0.0060 0.0035 -0.0697 -0.0574 0.0131 0.0072 0.1752 0.0868 0.0013 0.0087 0.3690 0.2277 0.0207 0.0141 Índices de Kakwani 1984 1989 0.3906 0.5441 0.0104 0.0140 0.5612 0.6438 0.0130 0.0153 0.3250 0.4996 0.0158 0.0162 0.1311 0.3566 0.0228 0.0195 0.0615 0.0028 -0.1237 0.0052 0.2264 0.0078 0.4786 0.0116 0.0648 0.0027 -0.1136 0.0050 0.1983 0.0067 0.4720 0.0108 1996 0.4616 0.0107 0.6467 0.0114 0.2967 0.0129 0.0444 0.0156 2002 0.4436 0.0054 0.6219 0.0068 0.3101 0.0082 0.0364 0.0118 Fuente: Elaboración propia con información de las enigh. Conclusiones En años recientes, la participación del gasto público en salud como proporción del pib destinado a la población abierta (cobertura universal) ha aumentado, de manera que se ha triplicado en relación con 18 años atrás (cuadro 2), y se ha intentado focalizar el gasto hacia sectores más necesitados mediante la descentralización en la administración y gasto de los recursos en el ámbito regional, en donde se han utilizado criterios de asignación de recursos con base en los indicadores de salud locales. Por tanto, el comportamiento de las transferencias en salud ha evolucionado de manera positiva en los últimos años, destacando sobre todo la cobertura proporcionada por los servicios del la ss. De hecho, estos servicios son los responsables en gran medida de los buenos resultados del sector salud, ya que instituciones como el issste muestran un comportamiento más bien nulo en su cobertura de los estratos económicos más bajos. Por su parte, el imss también ha mejorado la progresividad de sus servicios, aunque en menor medida que los de la Secretaría de Salud. De acuerdo con el análisis del nivel de progresividad para los diferentes subsistemas públicos de salud, es evidente que la creación de un siste- Economía, Sociedad y Territorio, vol. xi, núm. 37, 2011, 729-757 751 ma público único proveedor de salud sería la solución para dirimir las diferencias existentes, ello conllevaría a un sistema más equitativo donde los estratos de ingreso más desfavorecidos se beneficiarían de manera más proporcional, ya que mejoraría la cobertura y desaparecerían las todavía amplias discrepancias en los montos de transferencias existentes entre los diferentes subsistemas, con las repercusiones que pudiera traer en la calidad del servicio. En el caso de un escenario falto de reformas, al menos se exige una creciente financiación del sistema ss con transferencias públicas, para expandir su cobertura y elevar la calidad de sus servicios, ya que se ha visto que la población más beneficiada por los servicios de salud de la ss se compone de los sectores más empobrecidos de la sociedad. Recapitulando: el sistema de salud necesita una reforma integral, para universalizar los servicios y unificar las diferentes instituciones públicas proveedoras de salud. De esta manera se eliminará la duplicidad de servicios, se aprovecharán las economías de escala y de ámbito y se ahorraran recursos. Así, se eliminarán de manera más contundente las diferencias en la equidad de acceso a los recursos sanitarios, entre áreas rurales y urbanas, entre clases sociales y entre regiones, se ahorrarán recursos y la duplicidad de esfuerzos y, lo que es más importante, se elevaría el nivel de bienestar general de la población. Anexo metodológico A.1. Curva de Lorenz Es uno de los instrumentos de análisis de la desigualdad de la renta más básicos que existen, pero también es uno de los más ilustrativos y prácticos, especialmente cuando se trata de comparar diferentes distribuciones de la renta f(y), ya que estandariza el nivel de renta para su medición. La idea básica es ordenar las unidades de ingreso por su nivel de magnitud, empezando por el nivel más bajo y graficando contra la proporción acumulativa de la población que le corresponde, en ambos casos, tanto el ingreso como la población han de normalizarse, de forma que tomen valores que van de 0 a 1. Así, si existiese un distribución del ingreso muy desigual, los cuantiles (porciones) poblacionales con mayor ingreso recibirán una renta más que proporcional al tamaño de su población, mientras que los cuantiles menores recibirán una renta menos que proporcional. De esta manera, ante el hipotético caso de una total igualdad en la distribución del ingreso, cada cuantil poblacional recibirá exactamente el mismo cuantil de renta que le corresponde, donde cada individuo tendría el ingreso promedio (μ) y la curva de Lorenz sería una línea de 45º. 752 Vargas-Téllez, C. O.: Progresividad del gasto público en salud... Figura a.i Fuente: Lambert P. (2001: 26). Es fácil comparar en la figura a.i dos distribuciones de ingreso familiar, la curva A es menos desigual que la B, ya que está más cerca de la línea de igualdad. En el caso de que la curva de Lorenz para una función continua sea L(p), 0≤ p≤ 1, el ingreso del primer 100p por ciento de unidades de ingreso es Nz∫z0 yf(y)dy = Nμ, de manera que la curva de Lorenz L(p) en el p-valor se define como: p = F (z) ⇒ L(p) = z dy ∫ yf (y) μ 0 (a.1) A.2. € Curva de concentración Cuando se grafican porciones de una variable x contra quintiles en la distribución de otra y, se llama curva de concentración y se compara con la curva de Lorenz, por lo que puede situarse por arriba o debajo de ésta, de modo que la diferencia entre ambas captura las diferencias en la clasificación de las distribuciones. Éstas son muy usadas en el análisis de los efectos distributivos de los impuestos y transferencias públicas. Debe decirse que en las curvas de concentración no se ordenan los valores de la variable en cuestión de menor a mayor, sino simplemente se mantiene el orden preestablecido de la variable de referencia, la cual en la mayoría de las veces es el ingreso. Economía, Sociedad y Territorio, vol. xi, núm. 37, 2011, 729-757 753 A.3. Coeficiente de concentración Se le llama al área que se encuentra entre la curva de concentración y la línea de 45º, de manera análoga a como lo hace el coeficiente de Gini. Sus valores se encuentran entre -1 para concentraciones sobre los ingresos más bajos de la variable en cuestión, y +1 cuando la concentración se acentúa en los estratos con más ingresos, mientras que 0 indica una distribución neutral, lo que implica que cada persona o decil presenta una misma participación en la variable. Así, para el caso donde se analizan los efectos que tiene una transferencia determinada sobre el ingreso, el coeficiente de concentración para el ingreso postransferencia es: 1 ∫ CY + B = 1 - 2 LY+B (p) dp 0 (a.2) Y para las transferencias es: 1 ∫ CB = 1 - 2 LB (p) dp 0 Analizando el efecto que una transferencia tiene sobre el ingreso, se define a la curva de Lorenz pretransferencia LB(p) como: z p = F (z) ⇒ L(p) = ∫ yf (y) dy μ 0 € Mientras que la curva de concentración postransferencia LY+B se define: z p = F (z) ⇒ LY+B (p) = (y)] f (y) dy ∫ [ y-b μ (1 - g ) 0 € (a.3) (a.4) La curva de concentración de transferencia LB se expresa: p = F (z) ⇒ LB (p) = z ∫ [b (y) μgf (y) dy 0 (a.5) € Donde la relación de las tres curvas es: Ly = gLB + (1 - g) LY+B (a.6) Lo cual muestra que la curva de Lorenz LY del ingreso pretransferencia es un promedio ponderado de las curvas de concentración de transferencias LB y del ingreso postransferencia LY-B. Por tanto, 754 Vargas-Téllez, C. O.: Progresividad del gasto público en salud... LY+B ≥ Ly ⇔ LB ≤ Ly (a.7) Esta desigualdad indica que para que el ingreso postransferencia sea más equitativo que el ingreso pretransferencia, la transferencia sobre el € ingreso € € deberá ser más equitativo que el ingreso pretransferencia A.4. Índice de Kakwani y progresividad La pregunta que surge ahora es cómo capturar esa nueva desproporcionalidad creada por una transferencia progresiva y su efecto redistributivo en el ingreso. En este sentido, un índice de progresividad tiene la ventaja de capturar en un número la estructura de transferencias b(y) y la distribución de la renta pre f(y) y post f ’(y) transferencias. La desproporcionalidad de un impuesto o transferencia progresiva se aprecia en la distancia existente entre las curvas LY y LB, de manera que a mayor separación entre ellas mayor será el efecto de desproporcionalidad. Esta idea se complementa con lo señalado por Musgrave y Tun (1948), para quienes entre más desigual sea la distribución del ingreso antes de una transferencia progresiva, más fuerte será su efecto ecualizador. De esta manera surge el índice de Kakwani, el cual mide la progresividad en términos de la separación de las curvas LY y LB, a través de un índice que calcula dos veces el área existente entre ambas curvas. 1 ∏ =2 K ∫ [L (p) - L (p) ]dp 0 Y B (a.8) También expresado como ∏K = GB - CY (a.9) Así, ΠK aumentará si la progresividad de una transferencia se incrementa para cada nivel de ingreso, mientras la distribución del ingreso pretransferencia permanezca constante, aumentando así la progresividad efectiva. Debe decirse que los límites de este índice dependerán de la desigualdad de la distribución pretransferencia, y es (-1+Gy ) el límite inferior y (1+Gy ) el límite superior. Economía, Sociedad y Territorio, vol. xi, núm. 37, 2011, 729-757 755 Anexo gráfico Gráfica i Distribución de las transferencias públicas en salud a las familias, 2002 Fuente: Elaboración propia con datos de la enigh-2002 y ensa. Bibliografía imss (Instituto Mexicano del Seguro Social) (2005), Población derechohabiente, Cuaderno de memoria estadística, imss, México. inegi (Instituto Nacional de Estadística, Geografía e Informática) (1986), Aspectos metodológicos de la Encuesta de Ingreso y Gasto de los Hogares de 1984, inegi, México. inegi (Instituto Nacional de Estadística, Geografía e Informática) (1991), Aspectos metodológicos de la Encuesta de Ingreso y Gasto de los Hogares de 1989, inegi, Aguascalientes. inegi (Instituto Nacional de Estadística, Geografía e Informática) (1998), Aspectos metodológicos de la Encuesta de Ingreso y Gasto de los Hogares de 1996, inegi, Aguascalientes. inegi (Instituto Nacional de Estadística, Geografía e Informática) (2004), Aspectos metodológicos de la Encuesta Ingreso Gasto de los Hogares de 1984, inegi, Aguascalientes. 756 Vargas-Téllez, C. O.: Progresividad del gasto público en salud... inegi (Instituto Nacional de Estadística, Geografía e Informática) (2004), Sistema de Cuentas Nacionales, inegi, Aguascalientes. insp (Instituto Nacional de Salud Pública) (2000), “Encuesta Nacional de Salud 2000”, insp, Cuernavaca. Lambert, P. (2001), The Distribution and Redistribution of Income, Manchester University Press, Manchester. Musgrave, Richard y Thin Tun (1948), “Progressive taxation in an inflationary economy”, Journal of Political Economy, 56, University of Chicago, Chicago, pp. 498-514. ocde (Organización para la Cooperación y el Desarrollo Económico (1997), Economic Survey of Mexico 1997, ocde, París. Primer Informe Presidencial, Ernesto Zedillo (1995), “Anexo Estadístico”, Presidencia de la República, México. Sexto Informe Presidencial, Ernesto Zedillo (2000), “Anexo Estadístico”, Presidencia de la República, México. Scott-Andretta, John (2000), Progresa: una evaluación preliminar, Centro de Investigación y Docencia Económicas, México. Scott-Andretta, John (2004), “Transferencias públicas ( y otros ingresos) en especie en la medición de la pobreza”, Documento de Trabajo núm. 301, Centro de Investigación y Docencia Económicas, México. Scott-Andretta, John (2004), “Desigualdad en salud y en los recursos para la salud en México”, Documento de Trabajo núm. 302, Centro de Investigación y Docencia Económicas, México. Shorrocks, Anthony (1983), “Ranking income distributions”, Economica, 50, Blackwell Publishing, Londres, pp. 3-17. Tercer Informe Presidencial, Vicente Fox (2003), “Anexo Estadístico”, Presidencia de la República, México. Economía, Sociedad y Territorio, vol. xi, núm. 37, 2011, 729-757 757 Recibido: 8 de diciembre de 2008. Aceptado: 2 de diciembre de 2010. César Octavio Vargas-Téllez. Es doctor en economía aplicada por la Universitat Autónoma de Barcelona. Actualmente es jefe de la División de Estudios Profesionales en la Facultad de Economía de la unam. Sus líneas de investigación son: pobreza, desigualdad, redistribución de la renta y transferencias públicas, e incidencia fiscal. Entre sus publicaciones destacan: “Purchasing Power Parity across Mexican Cities: A Data Panel Analysis”, Applied Economics, 40, Warwick, Londres, pp. 1-9 (2008); “Desigualtat i estat del benestar a Amèrica Llatina”, Diàlegs Revista d´Estudis Polítics i Socials, 9 (34), Fundación inehca, Barcelona, pp. 93-117 (2006).	499
https://openalex.org/W3115346411	OpenAlex	Open Science	CC-By	2,020	Contribution of bone-reverberated waves to sound localization of dolphins: A numerical model	Aida Hejazi Nooghabi	English	Spoken	8,003	12,659	Received 5 May 2020, Accepted 2 December 2020 Received 5 May 2020, Accepted 2 December 2020 Abstract – We implement a new algorithm to model acoustic wave propagation through and around a dolphin skull, using the k-Wave software package [1]. The equation of motion is integrated numerically in a complex three-dimensional structure via a pseudospectral scheme which, importantly, accounts for lateral hetero- geneities in the mechanical properties of bone. Modeling wave propagation in the skull of dolphins contributes to our understanding of how their sound localization and echolocation mechanisms work. Dolphins are known to be highly effective at localizing sound sources; in particular, they have been shown to be equally sensitive to changes in the elevation and azimuth of the sound source, while other studied species, e.g. humans, are much more sensitive to the latter than to the former. A laboratory experiment conducted by our team on a dry skull [2] has shown that sound reverberated in bones could possibly play an important role in enhancing localization accuracy, and it has been speculated that the dolphin sound localization system could somehow rely on the analysis of this information. We employ our new numerical model to simulate the response of the same skull used by [2] to sound sources at a wide and dense set of locations on the vertical plane. This work is the ﬁrst step towards the implementation of a new tool for modeling source (echo)location in dolphins; in future work, this will allow us to effectively explore a wide variety of emitted signals and anatomical features. Keywords: Dolphin’s echolocation, Numerical modeling, Reverberation, Correlation Available online at: Available online at: SCIENTIFIC ARTICLE Acta Acustica 2021, 5, 3 A. Hejazi Nooghabi et al., Published by EDP Sciences, 2020 https://doi.org/10.1051/aacus/2020030 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 is properly cited. Corresponding author: aida.hejazi_nooghabi@upmc.fr Aida Hejazi Nooghabi1,2,, Quentin Grimal2, Anthony Herrel3, Michael Reinwald4,5, and La Aida Hejazi Nooghabi1,2,, Quentin Grimal2, Anthony Herrel3, Michael Reinwald4,5, and Lapo Boschi1,6,7 1 Sorbonne Université, CNRS-INSU, Institut des Sciences de la Terre Paris, ISTeP UMR 7193, 75005 Paris, France 2 Sorbonne Université, Laboratoire d’Imagerie Biomédicale (LIB),CNRS 7371 – INSERM 1146, 75006 Paris, France 3 UMR 7179 C.N.R.S/M.N.H.N., Bâtiment d’Anatomie Comparée, 75005 Paris, France 1 Sorbonne Université, CNRS-INSU, Institut des Sciences de la Terre Paris, ISTeP UMR 7193, 75005 Paris, France 2 Sorbonne Université, Laboratoire d’Imagerie Biomédicale (LIB),CNRS 7371 – INSERM 1146, 75006 Paris, France 3 UMR 7179 C.N.R.S/M.N.H.N., Bâtiment d’Anatomie Comparée, 75005 Paris, France 4 / Department of Biomedical Engineering, School of Biomedical Engineering and Imaging Sciences, King’s C ondon, UK ngineering, School of Biomedical Engineering and Imaging Sciences, King’s College London, WC2R 2LS 5 Department of Zoology, University of Oxford, OX1 2JD Oxford, UK 6 Dipartimento di Geoscienze, Università degli Studi di Padova, 35122 Padova, Italy 7 INGV Istituto Nazionale di Geoﬁsica e Vulcanologia, Via Donato Creti, 12, 40100 Bologna, Italy 6 Dipartimento di Geoscienze, Università degli Studi di Padova, 35122 Padova, Italy 7 INGV Istituto Nazionale di Geoﬁsica e Vulcanologia, Via Donato Creti, 12, 40100 Bologna, Italy 7 INGV Istituto Nazionale di Geoﬁsica e Vulcanologia, Via Donato Creti, 12, 40100 Bologna, Italy Received 5 May 2020, Accepted 2 December 2020 1 Introduction depending on the direction it comes from; these differences are summarized in the “head-related transfer function” (HRTF). Behavioural experiments on humans suggest that each individual unconsciously learns to use its HRTF to localize sounds [5]. It is generally accepted that, in practice, the auditory system is very sensitive to certain prominent peaks and notches of the frequency spectrum perceived by the ears, or “spectral cues”, whose amplitude and location along the frequency axis, controlled by the complex shape of the pinnae, depend on the elevation of the source; a num- ber of studies have shown that up-down and front-back ambiguities are dealt with based on such spectral cues [4, 6]. Most literature concerned with the evolution of sound localization postulates that this task is accomplished by means of several well established auditory cues, at least in mammals [3, 4]. Firstly, the binaural or interaural time- or phase-difference (ITD or IPD): the delay between the arrival time of a sound at the two ears. Secondly, the binau- ral or interaural intensity or level difference (IID or ILD): the difference in the intensity of a sound, as perceived at the two ears. The locus of sources that can be associated to a given ITD and/or ILD is a cone, sometimes dubbed the “cone of confusion” [4]. Since humans and other species are known to distinguish different sources within such a cone, a third psychoacoustic cue must exist. In fact, it is clear from the physics of acoustic-wave propagation that the same signal interacts differently with our anatomy The ITD/ILD/spectral-cue model has been successfully applied to explain the results of behavioural experiments, and has found neurophysiological and neuroanatomical support across different species [4, 7]. At least in the case of humans, it has also been shown to have limitations that can be predicted theoretically and are conﬁrmed by Corresponding author: aida.hejazi_nooghabi@upmc.fr *Corresponding author: aida.hejazi_nooghabi@upmc.fr This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creat which permits unrestricted use, distribution, and reproduction in any medium, provided the original wo A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 2 behavioural experiments. 1 Introduction Namely, (i) since binaural cues play a fundamental role, subjects that are deaf on one side can be expected and are indeed observed to perform poorly in sound localization tasks [4]; (ii) normal-hearing subjects fail at determining the correct elevation of narrow-band sources: elevation is determined through the identiﬁcation of spectral peaks and notches, which requires that the emit- ted sound be broadband [8, 9]; (iii) because spectral cues are not as accurate as ITD and ILD, at least humans have been observed to be less sensitive to changes in source elevation than azimuth: according to experiments, their minimum audible angle (MAA, or the smallest detectable angular dif- ference between the locations of two identical sources of sound) for sources within the vertical plane is never less than 4, vs. ~ 1 on the horizontal plane [10]. important heterogeneity within the bone. Based on grays- cale values, we deﬁne a model of the skull’s mechanical properties, and numerically simulate elastic wave propaga- tion through it via a pseudospectral scheme [1]. Impor- tantly, bone heterogeneity, neglected in other similarly- minded numerical studies (e.g. [17, 18]) is accounted for in our simulations. While it is impossible (for a number of technical reasons that we discuss) to reproduce Reinwald et al.’s [2] experiment exactly, our results share all the important features of their data. 1 http://www.k-wave.org/ 2 Method In this section, we introduce the numerical tool we employed; we explain how we built a model of the skull’s mechanical properties based on a computed tomography scan and we describe the conﬁguration of our numerical experiment. Behavioural experiments indicate that odontocete ceta- ceans, which rely on sound and on their biosonar to carry out many essential tasks, do not suffer from (some of) the mentioned limitations. They are known to emit high- frequency clicks and burst-pulsed calls as well as lower fre- quency whistles. For example, the frequency range of echolocation signals emitted by short-beaked common dol- phins is between 23 and 67 kHz [11]. Despite the absence of pinnae, bottlenose dolphins have been shown to be equally sensitive to changes in the elevation or azimuth of signals similar to their echolocation clicks [12, 13]; they have also been shown to respond to clicks with a MAA of 0.7, superior to the localization accuracy of other studied mam- mals [3]. It has been suggested [14] that these results cannot be explained without invoking, within the auditory system of dolphins, a localization mechanism different from the ITD/ILD/spectral-cue model attributed to other species. 2.1 Numerical simulation toolbox We treat the skull as a purely elastic medium and model wave propagation via k-Wave1, an open-source toolbox which solves the coupled ﬁrst-order equations [1]: orij ot ¼ kdij ovk oxk þ l ovi oxj þ ovj oxi ; ð1Þ ð1Þ ovi ot ¼ 1 q orij oxj ; ð2Þ ð2Þ An essential factor in assessing this speculation is the analysis of the dolphins’ HRTF, and the information it might carry pertaining to source location. In an earlier study [15], our team has tackled this problem with an exper- imental approach, recording and analyzing mechanical waves propagating through a dolphin skull. Their experi- ments were performed using two different signals: a nar- row-band chirp and a sinusoidal burst, both with a central frequency of 45 kHz. Reinwald [15] concluded that the skull shape and structure does not give rise to promi- nent, location-dependent features in the HRTF spectrum; they found, on the other hand, that the skull transfer func- tion contains sufﬁcient information for sources to be pre- cisely localized by an algorithm based on Pearson’s correlation, equivalent to the so-called “time-reversal” scheme postulated by Catheline et al. [16]. where r denotes stress, v particle velocity, and the Ein- stein summation notation is adopted. k and l are the Lamé parameters and q is the mass density. The compres- sional and shear wave speed (a and b, respectively) are, a ¼ ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ k þ 2l q s ; b ¼ ﬃﬃﬃl q r : ð3Þ ð3Þ Equations (1) and (2) are solved for an elastic isotropic medium using the Fourier pseudospectral method. The lat- ter is based on the Fourier collocation spectral method for computing spatial derivatives and the ﬁnite difference method for time integration [1, 19]. Equations (1) and (2) are solved for an elastic isotropic medium using the Fourier pseudospectral method. The lat- ter is based on the Fourier collocation spectral method for computing spatial derivatives and the ﬁnite difference method for time integration [1, 19]. 2.2 Model of dolphin’s skull The frequency band of Reinwald et al.’s [2] data was severely limited by their experimental setup; additionally, it would be very difﬁcult, if possible at all, to carry out sim- ilar experiments on full heads, including soft tissues: yet, soft tissues might have a non-negligible impact on the HRTF and thus on localization. We here accordingly develop a model of elastic-wave propagation through a dol- phin head, and validate it by application on the same skull that was employed in the Reinwald et al. [2] experiments. We obtain high-resolution (voxel size = 111 lm) X-ray tomography images of the skull resolving small but possibly The ﬂawless skull of a young adult short-beaked com- mon dolphin (Delphinus delphis) of ~50 cm length and ~20 cm width, was scanned at the AST-RX X-ray tomogra- phy platform at the National Museum of Natural History in Paris, using a v\|tome\|x L 240-180 industrial micro CT scan- ner with voltage = 150 kV, current = 310 lA and exposure time of 333 ms. Data were reconstructed using the datos\|x software and exported into 4106 slices of 16-bit TIFF images in coronal view. The resulting images are in 1 http://www.k-wave.org/ A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 3 Figure 1. A slice of the CT scan images illustrating the mandible and teeth at the original voxel size (left) and the dowsampled processed image (right). Figure 2. Three slices of dolphin’s skull, where we deﬁne the trabecular and cortical bones. Figure 2. Three slices of dolphin’s skull, where we deﬁne the trabecular and cortical bones. Figure 1. A slice of the CT scan images illustrating the mandible and teeth at the original voxel size (left) and the dowsampled processed image (right). We next associate a value for the mass density and wave speed to each voxel. We convert the images from 16-bit to 8-bit, so that the gray scale values range from 0 to 255. We apply a self-calibration on CT values, so that the brighter regions are assigned higher values of mass density and wave speed compared to the darker regions with lower CT values. We set mass density to vary from 1001 to 2000 kg/m3 with 255 equal increments. Next, based on the density associated to each voxel, the voxel-wise values of a and b are calcu- lated based on Equations (4) and (5), respectively. 2.2 Model of dolphin’s skull The value of 0.3 was used for the Poisson ratio (m) [20]: grayscale which is representative of the mean attenuation of X-rays at each voxel. Their voxel size is 111 lm along x, y and z directions, which is large compared to the grid size envisaged for our simulations. We hence downsampled the images. Voxel size is an important parameter as it deﬁnes the spatial grid size in the simulations. Moreover, decreas- ing the resolution implies loss in the information on the structure. The gray values are important because they are the basis for associating elastic parameters to each voxel. This image processing step demands a compromise between the computational cost of the simulations and the degree of information on the physical structure of the skull that should be kept. In Figure 1 we show a slice with the original voxel size (i.e., 111 lm) and the 20-times increased voxel size (i.e. 2220 lm). grayscale which is representative of the mean attenuation of X-rays at each voxel. Their voxel size is 111 lm along x, y and z directions, which is large compared to the grid size envisaged for our simulations. We hence downsampled the images. Voxel size is an important parameter as it deﬁnes the spatial grid size in the simulations. Moreover, decreas- ing the resolution implies loss in the information on the structure. The gray values are important because they are the basis for associating elastic parameters to each voxel. This image processing step demands a compromise between the computational cost of the simulations and the degree of information on the physical structure of the skull that should be kept. In Figure 1 we show a slice with the original voxel size (i.e., 111 lm) and the 20-times increased voxel size (i.e. 2220 lm). a ¼ 0:004q2 þ 14:9q 10664; ð4Þ ð4Þ b ¼ a ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ 1 2m 2ð1 mÞ s : ð5Þ ð5Þ We chose to implement this level of resolution, as a good compromise between accuracy and computational cost, after a suite of simulations on two-dimensional structures of comparable complexity; these tests showed that signals modeled at the receiver locations were not modiﬁed signiﬁ- cantly by changes in numerical grid spacing (See Appendix A for more details). Downsampling blurs the images and hence some detailed information on the structure is lost. 2.3 Numerical set-up In the experiment performed by [2], the skull is immersed in water far from the walls and ﬂoor of the pool, and is illuminated by sources along different elevations. The acceleration is measured using accelerometers attached to the posterior part of the mandible. In order to be close to the experimental set-up in [2] and since one of the goals of this study is to investigate the mechanism by virtue of which the elevation of a sound source is determined, in the following we shall position numerical sources on the me- dian plane (the hypothetical plane that divides the head into left and right), at a range of different elevations. In practice, we illuminate the skull with a plane wave whose wave vector varies from 0 to 90. Angle zero corresponds to the wave coming from the front of the skull while angle 90 corresponds to the wave coming from above it. Figure 5. Simulation conﬁguration. The solid curve marks the range of elevation of the sources. From each elevation, a plane wave is emitted. Figure 5. Simulation conﬁguration. The solid curve marks the range of elevation of the sources. From each elevation, a plane wave is emitted. in computational cost. We therefore adopt another approach here assuming that a plane wave already formed outside the simulation box, we consider the points located on the edges of the simulation domain as point sources, each emitting the same signal with a previously-calculated delay. By ﬁring an identical signal with the proper time delay from neighboring points, we ensure the formation of a plane wave front which in contrast to the point-source approach does not require a large simulation box. The source mask in 3D is two perpendicular planes at the edge of the simulation box where each point on these planes acts as a source. The input signal is a sinusoidal burst with the central frequency of 45 kHz and total duration of 100 ls as shown in Figure 4. Our simulation domain consists of a box of 0.3 m by 0.8 m by 0.7 m with the spatial grid size of 2.22 mm in all three directions which ensures a minimum number of 20 points per wavelength. The total number of grid points are 135 by 360 by 315. 2.2 Model of dolphin’s skull Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 4 0 0.05 0.1 0.15 0.2 0.25 0.3 time(ms) -1 0 1 Amplitude Figure 4. Signal (“sinusoidal burst”) emitted by sources in numerical simulations. Figure 5. Simulation conﬁguration. The solid curve marks the range of elevation of the sources. From each elevation, a plane wave is emitted. Figure 4. Signal (“sinusoidal burst”) emitted by sources in numerical simulations. 2.3 Numerical set-up In order to guarantee the stability of the simulations, based on the Courant–Friedrichs–Lewy stability criterion [19], we select c0 t x = 0.1, where Dt and Dx are the temporal and spatial steps, respectively. The value of 0.1 was chosen after careful trial and error; this value, sensibly smaller than 1, is compatible with example values suggested in the k-Wave documentation. Our numer- ical conﬁguration is shown in Figure 5. We record signals inside the pan bone (i.e. the thin pos- terior end of the lower jaw bone) on either side of the mand- ible. The exact locations of receivers were not recorded when conducting the analogue experiment [2], but the dis- tance between receivers was: in our simulations, receivers were positioned as similar to the experiment as possible, based on this datum and on several available photos of the experimental set-up. A 2D view of the position of the receivers is shown in Figure 6. Once the elastic parameters and the numerical proper- ties are determined, we illuminate the skull by a plane wave. This is equivalent to limiting our application to sources or echolocation targets that are not in the immedi- ate vicinity of the subject. It is important to note that we do not intend to model the actual acoustical environment of the dolphin, but to characterize its localization abilities. This is usually done by considering sound coming from dif- ferent directions, i.e., the angle of the incoming wave should be clearly deﬁned. Plane waves are better suited for this goal, since they have a unique and well-deﬁned direction of propagation. The receivers are positioned on the bone with identical elastic properties on each side and they measure velocity of oscillation along x, y and z directions. Each simulation takes approximately 24 h on CPUs of type Inter(R) Xenon(R) CPU E5-2695 v3. 2.2 Model of dolphin’s skull For example, the distinction between the trabecular and cortical bone becomes less obvious. These two main parts of the bone structure are shown for different slices with the original resolution in Figure 2. Equation (4) was obtained by ﬁtting to experimental data (compressional wave speed and mass density for human cortical bone samples). This data was obtained as described in [21]. Based on the Resonant Ultrasound Spectroscopy (RUS) method, anisotropic velocities are measured for cuboid specimens. As resonant frequencies are directly related to the dimension of the sample and the wave speed inside it, RUS provides values for anisotropic velocities independent of density measurements. Equation (4) was obtained by ﬁtting to experimental data (compressional wave speed and mass density for human cortical bone samples). This data was obtained as described in [21]. Based on the Resonant Ultrasound Spectroscopy (RUS) method, anisotropic velocities are measured for cuboid specimens. As resonant frequencies are directly related to the dimension of the sample and the wave speed inside it, RUS provides values for anisotropic velocities independent of density measurements. Figure 3 shows the map of the wave speeds for the same slice shown in Figure 1. Considering that the background medium is water with a = 1500 m/s and b = 0, the mini- mum and maximum values for the compressional wave speed in these maps are 1500 m/s and 3861 m/s and the corresponding values for b are 0 m/s and 2919 m/s. Also, the gray scale values vary from the original ones. It should be noted that the processed image here has been sharpened in order to magnify the contrast of the gray val- ues in the downsampled image and all the image processing steps have been performed using ImageJ software. (b) 0 500 1000 1500 2000 2500 shear wave speed (m/s) (a) 1500 2000 2500 3000 3500 compressional wave speed (m/s) Figure 3. (a) Compressional and (b) shear wave speed values for a slice of the computed tomography scan in a water background. (b) 0 500 1000 1500 2000 2500 shear wave speed (m/s) (b) (a) Figure 3. (a) Compressional and (b) shear wave speed values for a slice of the computed tomography scan in a water background. re 3. (a) Compressional and (b) shear wave speed values for a slice of the computed tomography scan A. 3.1 Waveform comparison Figure 7. Comparison of the acceleration signals recorded in the experiment (black) and the simulation (blue) for a source elevation of 25 (a) with the normalized amplitude and (b) with the scaled normalized amplitude. Dashed and solid arrows mark the direct arrival and coda windows, respectively. We adopt different criteria to validate the reliability of our numerical model of the skull. In all cases, we use data from the mentioned laboratory experiment conducted by our team on the same skull [2]. In this experiment, the skull was immersed in water, centered in depth and width inside a water tank and illuminated from different angles on the vertical plane using a broadband marine transducer. The input signal in the simulation and experimental case are the same (Fig. 4). The receivers in the experiment are accelerometers. The position of the receivers in simulations is very close to the ones implemented in the experiment with a maximum difference of ~5 mm. In contrast to the experiment where data are available for a range of angles varying from 90 to 90, we have data for the range of 0–90. Figure 8. Comparison of the experimentally recorded (black) and numerically simulated (blue) acceleration, for source eleva- tion of 82. The amplitude of the reverberated part of the experimental data is scaled as in Figure 7b. Figure 8. Comparison of the experimentally recorded (black) and numerically simulated (blue) acceleration, for source eleva- tion of 82. The amplitude of the reverberated part of the experimental data is scaled as in Figure 7b. As a ﬁrst proxy, we directly compare the experimentally recorded signals with their numerically computed counter- parts. In the latter case, acceleration is computed from velocity by numerical differentiation, and only the x compo- nent, which is normal to the mandible and pointing out- wards from it, is considered (Fig. 5). This component is the closest one to what is measured in the experiment. Compared to the input signal in Figure 4, the recorded sig- nals (in both experiment and simulation) are perturbed, due to the interference of the incoming wave with the bone-reverberated one. Because wave propagation is faster in bone than in water, reverberated signal arrives almost as early as the direct one, and cannot be easily separated from it by visual analysis. This phenomenon can be more or less prominent depending on the angle of incidence. 3 Results In this section, we show the results pertaining to the validity of our model through different sets of comparisons with the experimental case; namely as direct comparison of the signals, measurement of the inter-receiver time differ- ence and correlation maps. In order to numerically generate a plane wave, one might deploy a point source in the far ﬁeld of the object, but this approach requires a large enough simulation box to satisfy the far ﬁeld condition, i.e. a signiﬁcant increase A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 5 0 500 1000 1500 2000 2500 shear wave speed (m/s) Figure 6. Receiver positions shown by red full circles on a slice of the skull with values of shear wave speed. The background material is water. 0 500 1000 1500 2000 2500 shear wave speed (m/s) Figure 6. Receiver positions shown by red full circles on a slice of the skull with values of shear wave speed. The background material is water. Figure 7. Comparison of the acceleration signals recorded in the experiment (black) and the simulation (blue) for a source elevation of 25 (a) with the normalized amplitude and (b) with the scaled normalized amplitude. Dashed and solid arrows mark the direct arrival and coda windows, respectively. 0 500 1000 1500 2000 2500 shear wave speed (m/s) Figure 6. Receiver positions shown by red full circles on a slice of the skull with values of shear wave speed. The background material is water. Figure 6. Receiver positions shown by red full circles on a slice of the skull with values of shear wave speed. The background material is water. 3.2 ITD The second criterion for validating our numerical model is the time difference between the signals recorded at the left and right receivers. Assuming that the skull is symmet- ric, the interaural time difference (ITD) between the two received signals from the sources located on the median plane is expected to be zero. However, the dolphin skull is not symmetric. Here, we compare the values of ITD for sim- ulated and experimental signals. We ﬁnd the ITD by taking the difference between the arrival time of the ﬁrst peak in the signals recorded at the left and right receivers. This arri- val time was ﬁrst identiﬁed automatically, as the time when signal amplitude would exceed a selected threshold, and subsequently checked visually by the ﬁrst author of this article. This approach is applied to both simulation and experimental data, and the ITD versus the incidence angle for both cases is shown in Figure 9. C ¼ Pt¼tf t¼ti s1 tð Þs 0 2ðtÞ ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ Pt¼tf t¼ti s1 tð Þ2 q ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ Pt¼tf t¼ti s 0 2ðtÞ 2 q ; ð6Þ ð6Þ where s1(t) is a signal received at a given receiver from a given angle and s 0 2ðtÞ is the delayed version of s2(t) which could be any of the signals received at the same receiver from all other angles [23]. We prefer the combination of moveout and correlation rather than simple cross-correlation, because we have neglected the attenuation in our simulations. This results in relatively large-amplitude reverberations, which might cause cross-correlation to be amplitude-biased. Conse- quently, the maximum of the cross-correlation which is expected to be the maximum of the similarity could be biased. As can be seen, the variations of ITD values obtained from our simulations show the same overall “bias” as the experimental data, but with elevation-dependent ﬂuctua- tions that are less-reproduced compared to the experimen- tal case. Furthermore, an increasing trend from 0 to 20 is observed in the simulation data but it is not as sharp as the experimental case. It should be noted that the orien- tation of the incoming wavefront with respect to the skull in the experimental case is not as accurately-deﬁned as in the simulations. Therefore, there might be an error in the direc- tion of the incident wave as recorded by the experimenters in [2]. 3.1 Waveform comparison As an example, in Figure 7a, we show the comparison between the experimental and simulation data for the left receiver and an incoming angle of 25. experimental signal and show the resulting waveforms in Figure 7b. Amplitudes are scaled assuming an exponential decay of the signal with time (t), as is commonly done in medical ultrasound imaging. This technique, called time gain compensation (TGC), assumes an exponential decay of the signals as they penetrate into the attenuating tissue. In order to retrieve the lost energy by time, the amplitude is multiplied by exp(at). In our case, the value of a is chosen by trial and error. The consistency between the simulated and experimen- tal signals becomes less clear for some angles. The worst case of the observed vs. simulated waveform decorrelation is shown in Figure 8. In this case, the codas of the two sig- nals are not in phase, but, again, the envelope of the rever- berated waveﬁeld is captured fairly well. Simulated and experimental data for both left and right receivers are com- pared in Appendix B for a suite of different elevations of the source. Figure 7 shows that experimental and numerical signals are most often in phase with one another (this is true for both the direct and reverberated signal); the amplitude of experimental data is, however, strongly attenuated, while attenuation has been neglected in simulations. In order to facilitate the comparison, we scale the amplitude of the A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 6 Figure 9. Comparison of the time difference between the signals received at the left and right receivers obtained from the experimental (red) and simulation (blue) data. Uncertainty of the experimental values is around 5 ls. with unique target locations. The dolphin would then locate the source of the perceived click at the location correspond- ing to the stored echo that, of all those available in the library, is best correlated with it. In this case, we have two receivers but localization based on correlation is still possible thanks to the presence of a complex structure i.e. the skull. This is equivalent to what has previously been shown in time reversal experiments [22]. We quantify the effectiveness of this algorithm by using signals resulting from our simulations. This allows us to identify the contri- bution of different parts of the signal to localization. Figure 9. 3.2 ITD The slightly higher values of experimental ITD are likely to be related to the fact that the distance between the receivers in the simulations is ~5 mm shorter than in the experimental case. After aligning the signals, we explore two different approaches: We correlate only the direct part of the signals. Direct waves are those part of the signals that mimic the form of the incident wave that has not undergone any reverberations (see Fig. 7b for distinction between different contributions in signals). ) We apply the correlation to the full-time signals, (i.e. direct + reverberated waves) In the following, similar to Figure 9 in [2], we show the resulting correlation maps for the left and right receivers. The y axis in these maps is the true elevation of the source (/0) (see Fig. 5) and the x axis is the elevation of all sources available in the library (ai). The library includes the record- ings made at both receivers, and ai is the source eleva- tion for each pair of signals. The color values are representative of the maximum of the normalized correla- tion coefﬁcient. 3.1 Waveform comparison Comparison of the time difference between the signals received at the left and right receivers obtained from the experimental (red) and simulation (blue) data. Uncertainty of the experimental values is around 5 ls. Pearson’s correlation [23] is used as a quantitative mea- sure of similarity. In practice, our algorithm ﬁrst aligns a pair of signals based on the ﬁrst (non reverberated) arrival, similar to a “normal moveout” correction in seismics [24]. The correlation of the so aligned traces is then computed, summing over the sample by sample product of the two sig- nals. We then normalize the correlation value by the square root of the product of the energy of the two signals as follows, 3.3 Correlation-based localization A given biosonar signal, reﬂected by a target at given azimuth and elevation, will result in a similar received sig- nal at the ears of the echolocating individual. This signal importantly includes the coda arising from reverberations within the bone and is strongly dependent on target loca- tion. It can be hypothesized that, upon hearing a click-like sound, a dolphin could compare it with a library of echoes that, while training itself to echolocate, it has associated By construction, the correlation is maximum and equal to one only along the diagonal of the plots in Figure 10, and, A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 7 7 Figure 10. Correlation maps obtained from correlating only the direct arrivals for the (a) left and (b) right receivers vs the maps obtained from correlating the full-time waveforms for the (c) left and (d) right receivers for the input signal shown in Figure 4. Colorbar denotes the values of correlation coefﬁcient. Figure 10. Correlation maps obtained from correlating only the direct arrivals for the (a) left and (b) right receivers vs the maps obtained from correlating the full-time waveforms for the (c) left and (d) right receivers for the input signal shown in Figure 4. Colorbar denotes the values of correlation coefﬁcient. 0 0.05 0.1 0.15 0.2 0.25 0.3 time(ms) -1 0 1 Amplitude Figure 11. Signal (“linear chirp”) emitted by sources in numerical simulations. 0 0.05 0.1 0.15 0.2 0.25 0.3 time(ms) -1 0 1 Amplitude ideally, should quickly decay to zero away from the diago- nal. If correlation decays quickly to zero away from the diagonal, source localization resolution can be considered relatively high; conversely, large correlations for signiﬁcant differences between ai and /0 mean low resolution. When only the direct part of the traces is correlated, we observe that high values of correlation are spread over almost all angles, implying that the resolution of the localization is not high. On the other hand, when we correlate the full- time signals, there is a strong improvement in resolution, with the regions of high similarity becoming more concen- trated along the diagonal (i.e. true source elevation). This improvement is most striking in the 20–70 elevation range. We next changed the input signal to a linear chirp whose frequency varies between 20 and 45 kHz. The reason for choosing this form of input signal (shown in Fig. 3.3 Correlation-based localization 11) is to be as close as possible to the real world in the limits of the simulation conﬁguration. The correlation maps we obtained for this case (not shown here), look very similar to the case where the input signal is a sinusoidal burst. Figure 11. Signal (“linear chirp”) emitted by sources in numerical simulations. localization in time and space is achieved. The -3dB width of the focusing function is deﬁned as the width of the focus- ing pattern where the value of the correlation coefﬁcient falls to 70% of the maximum value (e.g. [2]). As we have data for a limited range of angles, and since the dominant improvement in localization happens for the angles between 20 and 70, we measure the 3dB width for this range of angles. The results of this exercise for the case of the chirp- like input are shown in Figure 12. This, again, reveals the increase in the precision of the localization as a result of taking into account the reverber- ated part of the signals. The same processing procedure applied to experimental data shows a similar enhancement in the localization. Moreover, the decrease in the 3dB width of the focusing function thanks to considering the This observation conﬁrms that information carried by signal reverberated in the bone enhances signiﬁcantly the precision of the localization. We can quantify this improve- ment through the width of the focusing function. The latter is common in time reversal experiments (e.g. [25, 26]), where through focusing of time-reversed received waves a A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 8 20 25 30 35 40 45 50 55 60 65 70 elevation of the source [°] 10 20 30 40 50 60 70 80 90 -3dB width [°] L-direct-sim L-full time-sim L-direct-exp L-full time-exp R-direct-sim R-full time-sim R-direct-exp R-full time-exp Figure 12. 3dB width of the focusing functions for the direct and full-time correlations and left (L) and right (R) receivers when the input signal is a linear chirp as in Figure 11 for simulations and a narrower-band linear chirp for experimental case. Figure 12. 3dB width of the focusing functions for the direct and full-time correlations and left (L) and right (R) receivers when the input signal is a linear chirp as in Figure 11 for simulations and a narrower-band linear chirp for experimental case. 3.3 Correlation-based localization reverberation part in the correlations, is consistent between the simulations and experiment for most elevations. model consists of only a part of the head and not the full head. Nevertheless, on the basis of this model we can con- clude that localization is indeed improved by information carried by bone. Having veriﬁed the validity of our model and the perfor- mance of our algorithm on this simpliﬁed model, we can now apply it to more realistic models and to a wide variety of species. The emphasis of this study was on a particular skull for which detailed experimental data are available, allowing the validation of our numerical model. The next, particularly important step will be to take into account soft tissues, which is necessary if any inferences relevant to biol- ogy are to be made from our results. Our future work will shed further light on the nature of sound localization in cetaceans, and its implications for evolutionary biology. 4 Discussion In this study, the propagation of elastic waves through a dolphin skull is modeled numerically, based on high-resolu- tion X-ray tomography of the skull, and a numerical algorithm that solves ﬁrst-order coupled equations for the velocity and stress in the time domain. We validate our numerical results with data from an experimental study [2] conducted on the same skull. We do not expect to repro- duce experimental data exactly, because: (i) our current numerical setup neglects attenuation (which is hard to quantify); (ii) there is an uncertainty in how we associate wavespeed and density estimates to the X-ray scan; (iii) position and orientation of sensors in the laboratory exper- iment also carry an uncertainty; (iv) the coupling between sensors and bone in the experiment is probably not perfect, but cannot be modeled easily in our framework. Taking all these differences into account, we consider the match between numerical and experimental results to be satisfactory. Conﬂict of interest Author declared no conﬂict of interests. Appendix A Effect of numerical grid spacing on recorded signals 9. J. Blauert: Sound localization in the median plane. Acta Acustica United with Acustica 22, 4 (1969) 205–213. Here, we explain the impact of downsampling of the CT images on the received signals. In order to gain time and for the sake of simplicity, we perform 2D tests in a conﬁgura- tion where the slice is put in water and is illuminated by a plane wave. The receivers are located on the opposite side of the slice. The conﬁguration is shown in Figure A.1. 10. E.A. Lopez-Poveda: Why do I hear but not understand? stochastic undersampling as a model of degraded neural encoding of speech. Frontiers in Neuroscience 8 (2014) 348. 11. M.S. Soldevilla, E.E. Henderson, G.S. Campbell, S.M. Wiggins, J.A. Hildebrand, M.A. Roch: Classiﬁcation of risso’s and paciﬁc white-sided dolphins using spectral prop- erties of echolocation clicks. The Journal of the Acoustical Society of America 124, 1 (2008) 609–624. Unlike our 3D simulations, in this conﬁguration we placed the receivers in water and not in the bone. The rea- son is that when the resolution is changed, the location of receivers would change, which introduces an artifact in the comparison. We repeat the simulation for three differ- ent levels of downsampling, i.e., 2 times, 10 times and 20 times downsampled with respect to the original resolution. The received signals are averaged over all the receivers and then normalized with respect to the maximum amplitude. The comparison of the received signals is shown in Figure A.2. y ( ) 12. E. Matrai, M. Hoffmann-Kuhnt, S.T. Kwok: Lateralization in accuracy, reaction time and behavioral processes in a visual discrimination task in an indo-paciﬁc bottlenose dolphin (tursiops aduncus). Behavioural Processes 162 (2019) 112–118. ( ) 13. D.L. Renaud, A.N. Popper: Sound localization by the bottlenose porpoise tursiops truncatus. Journal of Experi- mental Biology 63, 3 (1975) 569–585. gy ( ) 14. P.E. Nachtigall: Biosonar and sound localization in dolphins, in Oxford Research Encyclopedia of Neuroscience. 2016. 15. M. Reinwald: Wave Propagation in Mammalian Skulls and its Contribution to Acoustic Source Localization. Ph.D. Thesis, Sorbonne Université, 2018. Figure A.1. Conﬁguration of 2D tests for investigating the effect of downsampling. 16. S. Catheline, M. Fink, N. Quiefﬁn, R.K. Ing: Acoustic source localization model using in-skull reverberation and time reversal. Applied Physics Letters 90, 6 (2007) 063902. 17. Z. Song, Y. Zhang, T.A. Mooney, X. Wang, A.B. Smith, X. References 1. B.E. Treeby, J. Jaros, D. Rohrbach, B.T. Cox: 2014. Modelling elastic wave propagation using the k-wave matlab toolbox, in 2014 IEEE International Ultrasonics Symposium, pp. 146–149. We next employ our numerical data to evaluate the hypothesis, emitted by Reinwald et al. [2] that dolphin echolocation might involve the correlation of each newly received waveform with a library of recorded echoes. Our results conﬁrm that, in this approach, using the information carried by the waves that are reverberated through the bone before reaching the ear complex improves localization accuracy signiﬁcantly. 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Bioinspiration & Biomimetics 14, 1 (2018) 016004. 18. C. Wei, W.L. Au, D.R. Ketten, Y. Zhang: Finite element simulation of broadband biosonar signal propagation in the near- and far-ﬁeld of an echolocating atlantic bottlenose dolphin (tursiops truncatus). The Journal of the Acoustical Society of America 143, 5 (2018) 2611–2620. y , ( ) 19. B.E. Treeby, B.T. Cox, J. Jaros: k-wave a matlab toolbox for the time domain simulation of acoustic wave ﬁelds - user manual. 2012. 20. J.D. Currey: Bones, Structure and Mechanics. Princeton University Press, 2002. 21. X. Cai, H. Follet, L. Peralta, M. Gardegaront, D. Farlay, R. Gauthier, B. Yu, E. Gineyts, C. Olivier, M. Langer, A. Gourrier, D. Mitton, F. Peyrin, Q. Grimal, P. Laugier: Anisotropic elastic properties of human femoral cortical bone and relationships with composition and microstructure in elderly. Acta Biomaterialia 90 (2019) 254–266. Figure A.1. Conﬁguration of 2D tests for investigating the effect of downsampling. Figure A.2. Comparison of the signals for a slice with 3 different resolutions: 0.22 mm (red), 1.1 mm (black), 2.2 mm (red). 22. M. Fink: Time-reversal acoustics in complex environments. Geophysics 71, 4 (2006) SI151–SI164. ( ) 23. W.H. Press, S.A. Teukolsky, W.T. Vetterling, B.P. Flannery: Numerical Recipes in C. Cambridge University Press, 1992. 24. R.E. Sheriff, L.P. Geldart: Exploration Seismology. Cam- bridge University Press, 1995. 25. M. Fink: Time reversal of ultrasonic ﬁelds. I. Basic principles. IEEE Transactions on Ultrasonics, Ferroelectrics, and Fre- quency Control 39, 5 (1992) 555–566. 26. M. Fink: Time-reversal waves and super resolution. Journal of Physics: Conference Series 124 (2008) 012004. 27. R.L. Brill, P.J. Harder: The effects of attenuating returning echolocation signals at the lower jaw of a dolphin (tursiops truncatus). The Journal of the Acoustical Society of America 89, 6 (1991) 2851–2857. Figure A.2. Comparison of the signals for a slice with 3 different resolutions: 0.22 mm (red), 1.1 mm (black), 2.2 mm (red). A. Hejazi Nooghabi et al.: Acta Acustica 2021, 5, 3 10 Appendix B We observe that there is a good match in amplitude between the three signals in both the direct and reverber- ated parts. There is a slight difference in the phase of the signals which comes from the variation of elastic parameters from one model to the other. The important feature is that different wave packets are clearly captured even in the case of 20-times decreased resolution. Repeating this procedure for different slices, we observed a similar behaviour. This strongly suggests that our 3D simulations conducted on the 20 times downsampled images are sufﬁciently accurate to be compared to experimental data. This resolution pro- vides a geometry representative of the real geometry with sufﬁcient details on the medium and at the same time reduces the computational cost and time. Cite this article as: Hejazi Nooghabi A, Grimal Q, Herrel A, Reinwald M & Boschi L, et al. 2021. Contribution of bone- reverberated waves to sound localization of dolphins: A numerical model. Acta Acustica, 5, 3. Comparison of numerical and simulated signals In this appendix, we show visual and quantitative com- parisons of numerical and experimental results for a suite of different source elevations. The overall similarity between simulations and experiments is reﬂected in a value of the Pearson’s correlation coefﬁcients always >0.6 for the direct signal (Direct signals are found based on a threshold on the amplitude and the known duration of the input signal); cor- relation coefﬁcients for the reverberated signals are low, but visual analysis suggests that the envelope, if not the phase, of the reverberated signal is often reproduced fairly well (Fig. B.1). Figure B.1. Comparison between experimental data (black) and simulated data (blue) for different angles at (a) left and (b) right receivers. The correlation coefﬁcients between the two signals for both the direct (rd) and coda (rc) parts of the signals are indicated in the bottom left of each panel. Figure B.1. Comparison between experimental data (black) and simulated data (blue) for different angles at (a) left and (b) right receivers. The correlation coefﬁcients between the two signals for both the direct (rd) and coda (rc) parts of the signals are indicated in the bottom left of each panel. Cite this article as: Hejazi Nooghabi A, Grimal Q, Herrel A, Reinwald M & Boschi L, et al. 2021. Contribution of bone- reverberated waves to sound localization of dolphins: A numerical model. Acta Acustica, 5, 3.	5,842
52/tel.archives-ouvertes.fr-tel-02885002-document.txt_2	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	10,844	17,546	Soumis aux critères de véracité positiviste, ces postulats se voient tous invalidés. Seuls les postulats modernes, qui ont fondé l'épistémologie et ses critères discriminants, sont donc considérés comme justifiés 11 Nous préciserons particulièrement la notion de « paradigme » en introduction du premier chapitre. 29 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique 3.2 Le mythe comme refoulé scientifique dans la Modernité Longtemps dévalorisé comme un récit fabuleux et trompeur, le mythe énonce dans un langage imagé, mi-ésotérique, mi-populaire, les croyances d'une communauté. Il fonderait: les règles d'organisation et de conduite d'une société, ainsi que les représentations qu'elle a de ses capacités à transformer son milieu de vie. Autrement dit, « () le mythe structure le temps par rapport à une origine (la création du monde, la fondation d'une société) ou à un terme (le jugement dernier, l'utopie). » (Claval, 1998, p. 129) Chaque culture combine différents mythes dans une mythologie, rassemblés sans nécessaire cohérence, car les mythes sont élaborés à des époques différentes et leurs contradictions sont tolérées (Akoun et Ansart, 1999, p. 358). Les mythes connaissent une évolution par des réécritures variables selon les auteurs (le mythe d'Orphée selon Platon, Ovide), les supports de narration (oeuvre picturale, littéraire) et les époques (ex : de Prométhée à Frankenstein). Les variantes se différencient concrètement par une focale sur certains motifs mythiques, le recours à des figures esthétiques symptomatiques d'une époque, la référence à d'autres mythes Ainsi, -delà de la question de la croyance, le mythe est révélateur, par sa réactualisation permanente, de la société qui l'a créé et qui le véhicule. A cet égard, la réécriture des mythes s'inscrit dans la finalité d'un message à incarner qui serait particulièrement saillant pour la société. Nous différencions quatre interprétations du mythe selon le rôle social qui lui est accordé : une interprétation sociohistorique considérant les mythes comme la vérité de sociétés lointaines ou anciennes, une deuxième confinant le mythe au registre religieux ou sacral pour y chercher son universalité et son atemporalité, une troisième supposant la nature idéologique du mythe (de mystification ou de mobilisation), une dernière (que nous suivrons) qui - prenant en compte la nature moderne de notre situation d'analyse des mythes - pense le mythe comme un refoulé du discours scientifique positiviste. Le mythe comme récit dépassé par l'avènement des sciences modernes La première interprétation du mythe est sans doute la plus répandue, largement diffusée par l'histoire antique et l'anthropologie. Elle définit le mythe comme un récit de fondation aux traits fantastiques et héroïques s'opposant au Logos, entendu comme l'interprétation moderne des résultats de la perception. Cette définition moderne du mythe met à distance le mythe qui concernerait soit des sociétés anciennes (particulièrement la Grèce et la Rome Antique), soit des sociétés dites lointaines (inuit, indienne, nordique). Le mythe serait restreint à une société « suffisamment lointaine dans l'espace ou dans le temps pour que la science [moderne] n'ait rien à craindre d'une éventuelle comparaison » (Lagrange, 2012, p. 202). La pensée mythique serait une pensée « préscientifique », une forme d'explication du monde par l'intuition sensible (Bachelard, 1967). A cet égard, le mythe opérerait dans un registre différent de celui de la rationalité logico-formelle et, plus généralement, de la démonstration. Le mythe a notamment été mobilisé pour restituer les grands événements historiques, notamment les guerres entre des empires ; les récits mythiques constituaient une source pour les historiens de l'époque au même titre que des sources originales (Veyne, 1983). L'exemple le plus emblématique de ces mythes est sans doute l'allégorie de la caverne de Platon (1966) qui illustre la vertu d'apprentissage du mythe en tant que récit, tout en 30 Introduction soulignant ses limites comme source de connaissances exclusive et en appelant à ne pas réduire le monde de l'intelligible au monde sensible. L'allégorie de la caverne annonçait le scepticisme moderne vis-à-vis mythes tout en restant liée à sa forme narrative. La Modernité tend, en effet, à questionner les mythes sous l'angle de la vérité : est-ce que le récit mythologique était compris, pour les sociétés antiques, comme un récit réel qui assurerait une meilleure intelligibilité du monde? De ce questionnement sur la vérité, découle une relégation du mythe à une vérité partielle et altérée, inscrivant ainsi la connaissance dans une écriture progressive d'épurement mythologique. Comme le rappelle Paul Claval, « [o]n a cru longtemps que les sociétés rompaient avec la mythologie lorsqu'elles recevaient la révélation ou devenaient rationalistes. Le mythe n'y subsistait apparemment plus que dans le domaine de la fable, ou comme source de métaphores littéraires. » (Claval, 1998, p. 127). Pourtant, le mythe était loin de se restreindre au champ de la connaissance ou du divertissement, il construisait aussi à une réalité sociale. Par exemple, Jean-Pierre Vernant (1985) oppose l'âge du Mythos à celui du Logos, soulignant ainsi la modification lente et profonde des structures de la pensée dans la société et des relations sociales qui se nouent en fonction (ex : l'autorité sociale découlant du capital mythique de certaines familles « apparentées » à des héros mythologiques). Le mythe opérerait une fonction de « régulation sociale () en tant que production d'une communauté, réponses possibles à ses interrogations, ensembles de règles tacites et de valeurs intériorisées » (Lebreton, 2006). Le mythe comme récit structurant des affects et des conduites humaines Une autre acception essentiellement reprise en anthropologie des religions et du sacré présente le mythe comme un récit à portée explicative universelle et atemporelle, particulièrement véhiculée par Claude Lévi-Strauss, Mircea Eliade et Georges Dumézil (Dubuisson, 1993). Le mythe porterait sur les fondations psychologiques des affects et anthropologiques des conduites humaines. Ces auteurs cherchent des homologies structurales entre des mythes issuses de cultures différentes pour en déduire des invariants anthropologiques (Lévi-Strauss, 1974), psychologiques (Eliade, 1963) ou linguistiques (Dumézil, 1968). L'existence de ces invariants trouverait sa preuve dans la nature indiscutable des mythes : en effet, un mythe est identifiable comme tel par n'importe quel lecteur et ce, quelque soit sa culture (commune au mythe ou non) selon Claude Lévi-Strauss (1974). En posant le mythe comme une alternance de répétitions (anthropologiques, psychologiques et linguistiques) et de variantes (esthétique et narrative), ces auteurs supposent une continuité entre sociétés ayant créé un récit mythologique puis ses réécritures : celles-ci reprendraient toujours les mêmes motifs invariants qu'ils mettraient en récit et articuleraient différemment. Dès lors, les mythes étudiés sont essentiellement de nature cosmogonique : ils seraient le récit des origines ou de la création. Pour ce faire, le mythe est conçu comme un récit qui se réfère au passé, mais à une époque indéfinissable (ex : « Il était une fois »). Le mythe s'inscrit dans une temporalité dissociée du présent, mais « qui conserve dans le présent une valeur éminemment explicative dans la mesure où il éclaire et justifie certaines péripéties du destin de l'homme ou certaines formes d'organisation sociale. » (Girardet, 1986, pp.12 - 13). Opérer un détour par le mythe constituerait une solution temporaire pour un groupe socialement et historiquement donné de chercheurs pour pallier les oppositions ou 31 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique discontinuités de la pensée rationnelle, en soulignant des invariants irrationnels auxquels leurs sujets de recherche ne pourraient se soustraire : « l'objet du mythe est de fournir un modèle logique pour résoudre une contradiction » (Lévi-Strauss, 1974, p. 254). Dès lors, Claude Lévi-Strauss considère le mythe comme une architecture de l'esprit qui, considérant les limites de la pensée rationnelle à un moment donné, inclut une autre manière de penser qui serait universelle car reposant sur des dualités empiriques (cru et cuit, frais et pourri, nocturne et diurne) (Lévi-Strauss, 1962, p.36). De part son universalisme et sa préfiguration sensorielle, cette manière de penser constiturait une « pensée sauvage », inhérente à la nature humaine et préalable à la domestication par les sciences (ibid.) : car ils témoigneraient de l'universalité de certains affects et de leurs effets sur les comportements humains, les mythes s'imposeraient comme récits fondateurs et structurant des sociétés humaines. Afin de dévoiler ces structures universelles et atemporelles des sociétés humaines, les chercheurs s'intéressent aux traces laissées par ces invariants, qu'elles soient dans l'organisation sociale (Lévi-Strauss, 1974), les représentations des émotions (Eliade, 1963) ou la linguistique (Dumézil, 1968). Ils opèrent surtout par une approche comparatiste soit de mythes similaires peuplant des cultures différentes, soit de différentes écritures d'un même mythe. Claude Lévi-Strauss a développé une méthode d'analyse qui considère le mythe indépendamment de sa société d'écriture et peut donc être utilisée de manière systématique à l'ensemble des mythes cosmogoniques. En définissant le mythe comme un métalangage, il a dévoilé des outils narratifs d'évocation de symboles (ex : des figures de style), des unités de sens (« mythèmes ») associées chacune à un affect particulier, une structure narrative dont le déroulé des événements dépendrait des motivations affectives des conduites humaines. Les outils d'évocation symbolique et l'articulation narrative des mythèmes apparaissent comme les deux principaux leviers de variation mythologique. Or, ces outils sont choisis en premier dans le cadre d'une (ré)écriture mythologique, préfigurant alors vers certains affects. De même, l'articulation des mythèmes peut varier, avec comme conséquence une modification de la structure narrative (Claval, 1998). Mais, étant donné que les thèmes mythiques se caractérisent par la réversibilité de leurs « tonalités affectives et morales » (Girardet, 1986, p.17) 12, le caractère fictif et fantastique du mythe peut alors apparaître comme un atout : le mythe peut être pensé comme une animation créatrice qui assure une mobilisation des individus (Sorel, 1990). Le mythe serait pensé comme un instrument politique mobilisable aussi par les groupes subalternes, car il permettrait de construire une vision téléologique de l'action politique, d'identifier des groupes ou classes sociales à l'aune de cette vision, de conférer une mission à certains d'entre eux En tant que récit mettant en intelligibilité les phénomènes, leurs mécanismes et leurs causalités, le mythe pourrait être utilisé comme un outil de diffusion d'une idéologie. Le mythe se voit désigner une fonction dans la lutte politique, par sa puissance de mobilisation des énergies individuelles (Sorel, 1990). Plus précisément, il permettrait à un groupe spécifique de devenir acteur d'une histoire universelle en déployant une force à la fois créatrice et destructrice : ce serait un instrument d'action sur le présent. Dès lors, le mythe ne chercherait pas à refléter la réalité, mais s'en détacherait en tant que production autonome de la pensée qui viserait une transformation de la réalité. Ainsi, l'attribution d'une fonction politique au mythe conduit à l'inscrire dans une lecture dialectique : si le mythe est mobilisable comme un instrument d'une pensée révolutionnaire, la perspective marxiste lui confère aussi une capacité de mystification des classes dominantes. Georges Sorel identifie à cet égard un unique sujet des principaux mythes, antagonistes, du XIXème siècle : la « grève générale des syndicalistes et la révolution catastrophique de Marx » (Sorel, 1990, p. 36). Cette grève est considérée comme l'Événement par excellence, ce par quoi quelque chose de nouveau arrive, s'établit une rupture, une catastrophe, quelque chose comme un grand combat ou une bataille décisive. » (Zarka, 2009, p. 11). Du fait de cette dialectique, une méfiance perdure vis-à-vis du mythe qui est pensé, dans la lutte sociale, comme un intrument politique à déployer en dernier recours, dans les « situations désespérées » où la raison aurait échoué (Cassirer, 1993, p. 279). Le mythe comme refoulé de l'épistémologie positiviste Prenant le contrepied de l'interprétation sociohistorique et celle structuraliste, la dernière acception conçoit le mythe « moderne » comme un « refoulé » - c'est-à-dire un produit involontaire et dissimulé - des sciences modernes (Lacan, 1991 ; Marret, 2002 et 2006) et, plus précisément selon nous, de l'épistémologie positiviste. Les sociétés se voulant modernes prétendraient à une connaissance scientifique du monde qui évincerait le mythe : la science serait une affaire sérieuse qui ne pourrait revêtir la forme d'un récit aux traits fabuleux (Pudal, 1994). Le mythe est ici aussi entendu comme un instrument, mais pas de mystification ou de libération : il serait un instrument moderne pour rendre invisible certains sujets. Les questionnements contemporains sur la fonction fabulatrice ou la croyance dans les mythes témoigneraient, selon Paul Veyne (1983), d'une certaine interprétation du mythe comme l'ensemble des sujets dont le discours scientifique moderne refuserait ou ne pourrait pas 12 Raoul Girardet (1986) prend comme exemple les travaux de Gaston Bachelard sur les mythes relatifs à la maison qui évoquent un lieu de refuge, d'accueil et de sécurité, à l'inverse d'un autre espace clos qui est le cachot associé à un enfermement et une insécurité. 33 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique traiter : ces sujets concerneraient, selon lui, les croyances et l'Altérité. Ainsi, les contours des mythes dans les sociétés modernes définiraient les sujets dont l'épistémologie positiviste ne peut pas administrer la preuve. L'étude de ces myth nous renseignerait sur les sujets que la Modernité a invisibilisé, plus précisément sur les conditions et les conséquences de la domination du Logos (entendu comme l'émergence de la rationalité logico-formelle) selon Sophie Marret (2002). C'est par ce postulat que François Dosse (1991) explique l'intérêt de Michel Foucault pour la folie et son traitement psychiatrique (défini comme une science moderne) dans son analyse des formes pouvoir par la normativité. Etudier les pratiques associées à la folie et leurs traitements permettait à Michel Foucault de renseigner les conduites jugées déviante et les réactions de la société (sanction, réprobation, prohibition). Le mythe assurerait les conditions de reproduction de la Modernité face aux turbulences voire crises (sociales, écologiques). Tout d'abord, elle ne pourrait pas totalement s'en soustraire, car la reconnaissance de sa réalisation téléologique repose sur la diminution des croyances mythologiques. Ensuite, ne pouvant supporter des discontinuités dans son raisonnement du fait de son principe d'auto-fondation, elle puiserait dans les schèmes de penser mythologiques leurs forces de mobilisation et de persuasion, en cas de confrontation à des arbitrages échappant au régime de la rationalité logico-formelle. En effet, un schème de penser mythologique désigne des représentations qui forment une base de raisonnement, d'autant plus puissante qu'elle crée tacitement une adhésion : comme il est atemporel, universel, structurant et familier, le schème mythologique donne l'illusion d'un arbitrage rationnel. Ainsi, il semblerait que, si la Modernité n'a pas évincé les mythes, elle en a néanmoins circonscrit leurs champs d'intervention qui se dessineraient en filigrane de ceux de la Modernité : « le mythe représente une vision du monde cohérente et symétriquement opposée à l'époque moderne » (Dirschauer, 2005, p. 272). Dès lors, la nature du mythe évolue pour s'adapter à des sujets sur lesquels l'épistémologie positiviste ne bénéficie pas de « faits » à analyser : les conséquences en chaîne des actions et, plus généralement, le devenir des sociétés. Ainsi, alors que les mythes étudiés en anthropologie des religions et du sacré sont essentiellement de nature cosmogonique, les mythes modernes acquièrent une portée étiologique, c'est-à-dire de définition d'un terme à une société par sa mutation (ex : Progrès social) ou son effondrement (ex : collapsologie). Le mythe apparaît comme une forme d'intelligibilité à part entière, c'est-à-dire non diminuée par rapport à la pensée rationnelle, et singulière, c'est-à-dire de nature différente (Veyne, 1983). Acception reten e du mythe Dans cette thèse, nous suivrons cette dernière interprétation du mythe comme refoulé de l'épistémologie positiviste. Quatre raisons nous ont conduit à poser ce postulat : - l'urbanisme s'étant théorisé en période d'ancrage de la Modernité dans l'urbain (reposant sur des imaginaires urbains de l'émancipation collective) et d'extension aux sciences sociales de l'épistémologie positiviste (par la sociologie), la rupture épistémologique que signerait l'avènement du Logos ne s'est de fait pas appliquée aux mythes qui peuplent l'urbanisme (cf. acception 1) ; - l'urbanisme s'est construit comme un savoir opérationnel défini par une finalité de transformation matérielle, ce qui le différencie fondamentalement d'un récit cosmogonique aux traits fabuleux (cf. acception 2) ; - si le mythe est un instrument politique construit indépendamment des idéologies, alors le « mythe de l'urbanisme scientifique » (Dreyfus, 1969) n'aurait pas pu préfigurer le champ d'expertise de l'urbanisme par la qualification de ses savoirs (cf. L'étude des fondements mythologiques de l'urbanisme nous permettrait donc de renseigner les caractéristiques de l'urbanisme retenues comme légitimes par la professionnalisation. 3.3 Le rôle politique de la production de connaissances Nous posons comme dernier postulat le rôle une compréhension politique e de la production de connaissances. Loin du « grand partage » entre savoirs et actions revendiqué par la Modernité (Latour, 1997), nous posons la production de connaissances comme un mode d'action politique qui se situerait entre, d'une part, le renoncement et le désintérêt, et, de l'autre, l'action planifiée (Benasayag, 2006). Les professionnalités se situeraient dans cet entre-deux. Le savoir comme levier d'un pouvoir d'agir Poser ce postulat suppose de rompre avec l'idée d'une neutralité des savoirs (Berger et Luckmann, 1997). La connaissance serait un moyen de renouer avec un pouvoir d'agir qui nous aurait été confisqué par la société de l'hyper-information selon Miguel Benasayag (2006). Les savoirs concernés par cette capacitation ne seraient ni opératifs, ni instrumentaux, ni relatifs à la conduite de l'action. Il s'agit de savoirs théoriques, soit scientifiques soit produits par l'agir. En effet, en créant des représentations territoriales, les savoirs orienteraient les rapports des habitants à leurs lieux de vie ; en mettant en intelligibilité des phénomènes, les savoirs révéleraient de potentiels leviers d'action qui pourraient libérer une volonté d'agir ; en proposant des modèles théoriques, les savoirs préfigureraient des récits de transformation du monde (Barbier, 2001) Nous pensons que la production de connaissances dans les métiers relève tout autant de l'agir dans les territoires que des actions intervenant directement sur la matérialité urbaine, comme le projet urbain, la planification, la programmation Nous identifions trois modes d'action par la production de connaissances : - la mise en intelligibilité de phénomènes permettant d'anticiper conséquences donc éventuellement les redresser, et d'identifier leurs causes donc de potentiellement les reproduire ou les éviter ; - une suggestivité performative pour les théories scientifiques par l'autorité qui leur est conférée en dehors du milieu scientifique et universitaire, dès lors que des critères de scientificité sont annoncés ; Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique - une capacité de mobilisation par des théories sociales construites comme des récits voire des méta-récits, d'aurant plus forte que les sujets susciteraient un sentiment de concernement, comme les crises écologiques ou les transformations volontaires des lieux de vie par l'urbanisme (Karsenti, 2017). Précisons que, si la formalisation des savoirs et leur diffusion contribuent fortement à identifier la connaissance comme un mode d'action, elles ne constituent pas une condition catégorique. En effet, même si elle n'est pas diffusée, la production de connaissances sur le territoire, sur l'urbain, etc., oriente fondamentalement les pratiques futures du praticien : elle déterminerait les choix futurs de conduites et de registre instrumental d'action, d'identification des problématiques et des acteurs pour les résoudre Le politique dans la Modernité : raison versus sensible Nous nous différencions, à cet égard, de l'acception moderne du politique qui en fait une activité rartionalisante de séparation et de hiérarchisation des individus et des activités par l'érigement de frontières. La compréhension du politique que nous retenons s'oppose à celle défendue par le projet moderne : dans une société visant la Modernité, le politique opérerait une fonction de construction de frontières qui identifieraient des espaces hermétiques, et de ventilation d'un certain nombre d'activités dans ces espaces nouvellement créés. Trois frontières modernes enchâssées ont particulièrement retenu notre attention : l'opposition profession / recherche (ou théorie / action), l'opposition vie professionnelle / vie sociale et l'opposition raison / sensible. Si l'usage fréquent et abusif du qualificatif « professionnel » pour désigner l'urbaniste s'inscrit dans les stratégies de professionnalisation (en sousentendant l'existence a priori d'un groupe unifié et aux caractéristiques communes), il témoigne aussi de la prégnance de plusieurs partages modernes (Latour, 1997) traversant l'urbanisme et ses métiers. La pratique en urbanisme est entendue le plus souvent en opposition à la recherche scientifique supposant par là une épistémologie strictement théorisante (Doucet, 2014), et dans une lecture duale entre sociale et vie professionnelle (Dubar, Tripier et Boussard, 2015). Ces deux « petits partages », pour reprendre l'expression de Bruno Latour (1997), supposent l'existence d'un « Grand partage » fondateur de la Modernité : la différenciation des sphère sensible et rationnelle, qui pose cette dernière comme seul registre légitime de justification d'une délibération démocratique selon Jacques Rancière (2005). Au regard de cette définition moderne du politique, l'urbanisme se revendique d'une neutralité politique en tant qu'il ne participe pas aux délibérations politiques et suivrait des choix rationnellement fondés. Si « [l]e rapport semble très étroit entre les mots urbanisme et politique qui qualifient l'un comme l'autre l'organisation et la gestion de la cité » (Blais, 2000, p. 74), l'urbanisme se positionne néanmoins comme un savoir au service de stratégies politiques prises hors des métiers. Afin d'éviter de reproduire ces partages contre-productifs concernant les professionnalités, cette thèse demandera une vigilance dans le choix des termes. Si le qualificatif « professionnel » renvoie à l'exercice d'une activité rémunérée, nous essaierons désormais de l'employer essentiellement lorsqu'il fait référence au terme de « profession » entendue au sens anglo-saxon - qui est le plus restreint - comme une activité rémunérée dont l'organisation, les recrutements et l'accès au marché sont réglementés par un certain nombre de dispositifs ad hoc reposant sur des critères de maîtrise de compétences qui seraient propres à la profession. Cet usage restrictif des termes dans cette thèse nous permettra d'éviter certains allants de soi comme l'association de la professionalisation à l'opérationnel donc, en filigrane, l'éviction de la production de connaissances dans les métiers de l'urbain. Le politique comme mise en liens de mondes singuliers En opposition à cette définition moderne du politique, nous entendons ce dernier tel que définit par Jacques Rancière (1995 et 2000) comme une activité certes d'ordonnancement du monde mais opérant par la production d'une cohérence entre différentes unités de sens éparses. Plutôt que de créer des frontières hermétiques, le politique agencerait les activités et phénomènes en créant des ponts entre eux. Faire du politique reviendrait à reconfigurer les places et les fonctions des activités et des individus : c'est un « système d'évidences sensibles qui donne à voir en même temps l'existence d'un commun et les découpages qui y définissent les places et les parts respectives. Un partage du sensible fixe donc en même temps un commun partagé et des exclusives » (Rancière, 2000, p. 12). Concrètement, le politique opérerait par 1/ identification de groupes d'acteurs, d'intérêts, de similitude sociales, d'appartenance territoriale, etc., puis 2/ partition de ces groupes dans des liens de nature diverse (de dépendance, de réciprocité). Plusieurs chercheurs comparent les résultats du politique à une « grammaire » (Négri, 2006 ; Lemieux, 2009), c'est-à-dire un ensemble de règles « qui permet aux membres d'une communauté de juger correctement, c'est-à-dire de lier correctement à des discontinuités survenant dans le monde (corps, objets, matériaux, gestes, paroles) des descriptions et d'éprouver vis-à-vis de certaines de ces descriptions un sentiment d'évidence » (Lemieux, 2009, pp. 21 - 23). Selon Jacques Rancière (1995), le politique reposerait sur un mode d'action essentiellement langagier, aussi bien pour la mise en ordre d'une société que pour l'initiation d'un mouvement de révolte, en passant par les stratégies de reproduction sociale. Autrement dit, le politique s'exprimerait par des paroles, comme l'écriture d'une mémoire collective et l'énonciation de spéculations, la déclaration de revendications et la proclamation de droits, la protestation face à une injustice et l'identification d'un destin propre un groupe, la narration du récit d'une lutte et la grammaire de l'action d'une communauté politique Cette portée politique du Logos concerne certes la réification de théories politiques qui préfigurent les moyens d'action choisis, la reconnaissance dans un sort collectif, etc., mais elle relève surtout de la subjectivation, c'est-à-dire de la construction de sujets en politique. En effet, la mise en mots contribuerait à la prise de conscience de l'ancrage politique des expériences que nous vivons : elle permettrait de mettre en corrélations des phénomènes, d'identifer des causalités, de se remémorer des expériences La parole politique aurait la particularité de reposer, selon Jacques Rancière (2017), sur des méthodes d'argumentation issues de la littérature au sens large du terme. En effet, le récit nous assure une « puissance d'affecter et de transformer, liée au fait que les récits, comme l'anthropologie nous l'a appris, nous font littéralement tenir debout. » (Hache, 2014a, p.17). Cette proximité du politique et de la littérature explique, en partie, le rôle que peuvent jouer les schèmes mythologiques dans le urbain crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur c mythologique projet moderne pour dépasser les dictions ou combler les vacances de la rationalité totalisante moderne, comme nous le verrons dans notre première hypothèse. Or, cette activité d'ordonnancement du politique s'est vu complexifiée par un affaiblissement de l'adhésion aux méta-récits politiques ayant dominé au XXème siècle (Ion, 1997 ; Frère et Jacquemain, 2013). Par méta-récit nous entendons une « narration à fonction légitimante » (Lyotard, 1979, p. 34), considérant le terme de narration comme l'exercice de mise en forme par le Logos, le récit produit et son usage social. La capacité légitimante du méta-récit repose sur 1/ son cadre d'analyse qui est englobant voire totalisant des phénomènes et acteurs, 2/ son sujet d'analyse que sont les grandes structures sociales et rapports de domination, ainsi que 3/ son usage social du récit pour susciter un désir de mobilisation et préfigurer l'expérience (par anticipation de sa réalisation et de ses effets). Le méta-récit a une vocation symbolique à l'universel par l'identification claire de sa filiation politique (nom, symboles, valeurs) et d'un adversaire commun, la construction d'une vision téléologique de l'action et la production d'outils généralisables pour y parvenir L'affaiblissement de ces méta-récits se traduirait pas une évolution des formes d'engagement vers une « individuation » (Ion, 2012), c'est-à-dire des engagements situés, concrets, temporaires. Ces engagements seraient enchâssés dans des trajectoires personnelles et leurs dotations en connaissance, des situations quotidiennes et leurs leviers de bricolage ou de détournement fonctionnel Par là, ces engagements témoigneraient d'une désafiliation des individus aux structures d'appartenance héritées (ex : sociaux, familiaux, professionnels) qui, jusque-là, se posaient comme systèmes exclusifs et déterministes d'affiliation des individus à partir de rôles et de statuts devenus obsolètes ou partiels (ex : classes sociales, famille nucléaire). « Cela ne veut évidemment pas dire que l'individu deviendrait une sorte d'atome libre. Il reste toujours un individu défini par des processus sociaux, par son appartenance à une communauté langagière, par des institutions, etc. » (Ion, 2010, p. 78). C'est le caractère exclusif et primaire de ces systèmes d'affiliation qui est remis en cause ; ils restent pertinents et efficients pour une identification construite par les individus et non plus une affiliation primaire et exclusive. Une des conséquences de cette individuation, c'est en effet le passage d'identités d'appartenance à des identités de construction, par un « double affranchissement par rapport aux appartenances de type communautaire, et par rapport aux réseaux verticaux, fédératifs, inscrits dans les constellations idéo-politiques » (Ion, 2001, p. 23). Les savoirs dans les métiers : instruments ou modes d'action? Or, cette conception du savoir se révèle relativement absente de l'urbanisme, ce qui nous conduit à questionner par ce biais la place des professionnalités dans l'urbanisme. Une ligne de partage se dessine au sein des métiers en fonction de leurs rapports aux savoirs : dans certains métiers associés à la « fabrique de la ville » prédominerait une fonction instrumentale du savoir, tandis que d'autres investiraient la production de connaissance comme un espace d'expérimentation de leurs champs. En effet, en convoquant une fonction essentiellement instrumentale des savoirs (Faburel, 2017a), l'urbanisme se pose comme une activité d'application de savoirs supposés neutres et mis à disposition des politiques publiques et projets urbains (Choay et Merlin, 1989). Il s'inscrit dans « une forme de relation entre le savoir et l'action dans laquelle la théorie précéderait et déterminerait la pratique » (Guigue, 1998, p. 115), supposant par là : - une antériorité temporelle de la théorie et, en corollaire, une neutralité de la théorie conférée par son isolement de la vie active ; - un lien causal de la théorie à la pratique et, en corollaire, une réduction de la pratique à l'application linéaire et mécanique de préceptes théoriques. Cette acception des savoirs semble plus généralement s'inscrire dans une « épistémologie de la science appliquée » (Schön, 1994) qui sépare, d'une part, la production de savoirs et, de l'autre, l'action opérant au sein de la société urbaine par l'intervention matérielle ou la régulation sociale. Le savoir et l'action seraient deux activités relevant d'acteurs et de temporalités distincts, et répondraient de logiques et principes différents. Relativement présente dans les sciences sociales (Doucet, 2014), cette épistémologie a structuré la répartition des activités entre les métiers de l'urbanisme. La production de connaissances serait l'apanage de la recherche académique, de bureaux d'étude, des filières « & Développement » des groupes opérationnels et des services « Prospective » des collectivités ; elle serait réalisée sans lien avec l'action, ce qui lui assurerait une neutralité sur le sujet ; elle répondrait d'une logique d'amélioration de la connaissance délivrée aux divers acteurs de la ville (praticiens, associations, usagers) ; enfin, elle opérerait par une montée en généralité de cas d'étude ou par une analyse comparative, qui assurerait une posture distanciée à la société urbaine immédiatement proche. A l'inverse, l'action serait ancrée dans la société urbaine ce qui lui permettrait d'identifier des problèmes, dont les causes et les mécanismes seraient renseignés par les apports théoriques, et auxquelles des acteurs spécialisés répondraient. Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique Nos postulats nous renseignent sur les conditions théoriques de politisation des métiers par les professionnalités. Tout d'abord, relevons que les mutations du politique tendent à faciliter la reconnaissance des professionnalités comme geste politique. En effet, d'une conception relativement unifiée et stabilisée de ce qui ferait politique, nous cheminons vers une diversité de réalités, dépendant des expériences et des affiliations singulières des individus. Toutefois, poser la production de connaissance comme une activité politique suppose alors de rompre avec quelques partages épistémologiques modernes et d'engager un positionnement explicite sur la place (rôle social, fonction instrumentale) et la nature (discipline, épistémologie) des savoris dans les métiers (postulat 3). Ici, il semble difficile de concilier le projet politique de Modernité et une évolution des métiers par les professionnalités, car la Modernité tend à refouler les sujets contredisant sa téléologie progressiste ou ne pouvant être prouvés par son épistémologie positiviste. Elle leur confère un statut mythologique qui les invisibilise dans les arbitrages politiques (postulat 2). D'une manière générale, ce contrôle des savoirs par la Modernité s'explique par le rôle conféré aux savoirs scientifiques dans la réalisation du projet moderne. L'amélioration des savoirs assurerait un progrès humaniste de l'émancipation collective et un progrès social d'augmentation du confort. Paradoxalement, si l'épistémologie positiviste se réclame d'une dissociation forte du politique, elle a été produite en vue de permettre aux savoirs scientifiques d'assurer leurs fonctions humaniste et sociale - définies par le projet politique de Modernité (postulat 1). 4. Hypothèses : les professionnalités de l'urbain dépassement de la dialectique Modernité / Mythologie comme Les liens entre mythe et urbanisme ne constituaient pas notre sujet de thèse initial (les professionnalités), mais se sont imposés à nous comme cadre interprétatif de nos matériaux de recherche. Nous avons été confrontés à la même situation qu'Olivier Boulba-Olga et Michel Grossetti (2018) : l'observation d'un discours sur le régime de l'évidence naturelle, c'est-àdire soumis à aucune justification scientifique, affective ou politique. Lors de l'analyse de la littérature sur les métiers, nous avons relevé des principes d'action (mythe de Prométhée), des représentations territoriales (mythe de Babel) et des postures d'expertise (mythe d'Orphée) récurrents, posés comme des certitudes mais ni empiriquement fondés, ni énoncés comme un portage politique. Ces certitudes se sont pourtant révélées questionnées par les praticiens rencontrés, au regard des crises écologiques, car elles ne correspondaient pas à l'évolution contrainte, expérimentée ou espérée, de leurs métiers. Notre première hypothèse est née de cette observation : l'urbanisme s'inscrirait dans une ambition moderne et, à cet égard, serait traversé de schèmes mythologiques correspondant à des arbitrages politiques refoulés par l'épistémologie positiviste. Nous pensons que « [l]a modernité recèle en son sein une part grandissante d'irrationalité qu'elle refoule et dénie sous les oripeaux de la rationalité technicienne ou encore d'une rationalité tronquée 13 qui se limite à « l'apologie du calcul efficace ». » (Frémaux et Guillaume, 2014, p. 126). Cette première hypothèse expliquerait l'investissement de la littérature scientifique pour délimiter les contours des métiers, car la Modernité conf un rôle primordial aux savoirs scientifiques dans la justification de l'action (Lévy-Leblond, 2000). Elle expliquerait aussi l'enlisement des débats par défaut d'arbitrage sur la nature de l'urbanisme (discipline, champ d'action, savoir de gouvernement), car la Modernité ne reconnaît que la rationalité logico-formelle comme régime de délibération démocratique (Rancière, 2005). Or, les crises écologiques viendraient perturber, le dévoilant par là, cet équilibre entre refoulement et schème mythologique. Notre deuxième hypothèse porte sur la fragilisation de la prétention d'auto-fondation de l'urbanisme par la résurgence de ses externalités et refoulés. Les crises écologiques dévoileraient des sujets sur lesquels l'urbanisme opère des arbitrages politiques dissimulés par des schèmes mythologiques. En dévoilant des sujets politisés dans les métiers, les crises écologiques créeraient des espaces du possible pour l'expression des professionnalités. Notre troisième hypothèse pose la capacité de transformation des métiers par la diversification et l'hybridation de leurs savoirs. 4.1 Première hypothèse : la construction l'urbanisme par sa légitimation moderne mythologique de L'urbanisme s'est construit et différencié dans le champ de l'urbain par un rapport fort à la technique et une structuration scientifique des activités (Lefebvre, 1961 ; Dreyfus, 1969 ; Claude, 2006 ; Beaudet, 2007). Les sciences modernes y ont été convoquées comme un gage légitimaire (Faburel, 2017a). Or, nous avons vu que, malgré des prétentions à l'autonomie et la certitude, ces sciences reposent sur une épistémologie dont les principes revêtent un « caractère fictif » (Pudal, 1994), voire « mythique » (Claval, 1998 ; Mucchielli, 2004). L'urbanisme n'échapperait donc pas à ce fondement mythologique, largement dénié par un discours de justification scientifique. Plus précisément, l'urbanisme se composerait d'une « constellation mythologique » c'est-à-dire d'une trame continue et articulée de différents mythes. La prétention scientifique de l'urbanisme : un réductionnisme des facteurs de choix Au regard du rôle conféré aux sciences modernes dans sa légitimation, l'urbanisme semble reproduire la dialectique entre Modernité / Mythologie, largement déniée par un discours de justification scientifique mais assurant des conditions de reproduction de la Modernité par le refoulement de certains sujets. En effet, Jacques Dreyfus critique ce qu'il nomme le « mythe d'un urbanisme scientifique » (1969, p. 488) dans la mesure où les théories, les outils et les modèles urbanistiques bénéficieraient d'une justification scientifique : ils reposeraient sur des idées conférées au rang de vérité au nom de leur nature scientifique annoncée. Ce mythe 13 Le mot souligné est mis en exergue (italique) dans le Profession nalités de l'ur bain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique fonderait une « rationalisation de la décision », rédu isant la dé cision à des arbitrages quantifiables répon dant de beso ins fonctionnels ( . 491). Cette rationalisation peut aussi bien orienter les décisions portées par des acteurs de l'urbanisme (Offner,1993) que l'interprétation des pratiques et des usages habitants (Chombart de Lauwe, 1965). Ainsi, Jean-Marc Offner (1993) relève une « mystification scientifique » autour du principe de l'effet structurant des transports qui - bien que souffrant de critiques théoriques et, surtout, infirmé par des études empiriques - se révèle fortement mobilisé par des chercheurs, des praticiens et des décideurs de l'urbanisme. De même, Paul-Henry Chombart de Lauwe reproche à l'urbanisme une vision strictement rationalisante des actions humaines, donnant lieu à l'identification d'une logique d'homo oeconomicus, à l'opposée d'« une anthropologie dans laquelle seraient définies les aspirations des hommes d'aujourd'hui en fonction de l'avenir qui s'impose à eux, en fonction des valeurs auxquelles ils sont attachés, des croyances, des symboles, des mythes qui orientent leur pensée » (1965, p. 73). En somme, l'homologie structurale entre un schème mythologique et le raisonnement urbanistique permettrait aux métiers de s'affranchir de l'administration de la preuve pour légitimer, au nom d'une rhétorique scientifique de causalité linéaire, certains principes d'action de l'urbanisme et de l'aménagement. S'intéresser aux mythes dans le champ de l'urbanisme revient donc à s'intéresser à ce / ceux que l'urbanisme a exclu par sa prétention moderne à une délibération strictement rationnelle, au statut de ces exclus (ignorés, impensés, refoulés), ainsi qu'aux mécanismes de construction de cette exclusion (neutralisation de la parole dans la scène discursive, rhétorique exclusive de la légitimité, dévalorisation par le recours à certains imaginaires, recul des frontières de l'urbain). Dès lors, nous pouvons interroger les réactions qui naitraient du dévoilement de ce fondement mythologique de l'urbanisme : déni, négation, resserrement autour de certaines théories historiques, cristallisation autour d'imaginaires professionnels historiques Le mythe en urbanisme : un discours illusoire voire trompeur Au regard de la prétention scientifique de l'urbanisme, il n'est pas onnant que l'acception retenue du mythe retienne un décalage avec le réel. L'interprétation du mythe comme un récit de fondation socio-géographique d'une société est celle la plus souvent retenue dans les champs de l'urbanisme (ex : Cabantous, 2004 ; Salomon-Cavin, 2005) et, dans une moindre mesure, de la géographie (ex : Berdoulay, Castro et Gomès, 2001 ; Guillaud, 2008). En délimitant le lieu narratif du mythe, l'urbanisme et la géographie lui assignent une fonction de structuration sociale d'un espace, notamment de cohésion sociale ou de fédération des identités individuelles dans des images et / ou symboles spatiaux. Par exemple, le mythe du « bon sauvage » a joué un rôle essentiel dans l'appropriation des Alpes par la société urbaine du XVIIIème siècle, puis le développement du tourisme (Salomon-Cavin, 2005). Le mythe associé à un espace urbain renseignerait sur les caractéristiques sociales d'une forme urbaine : - soit en dévoilant ses capacités de cohésion, pour l'espace public (ex : Berdoulay, Castro et Gomès, 2001) ou les instances sociales de proximité (Genestier, 2001) ; - soit en soulignant la rupture qu'elle instaurerait entre des formes jugées classiques (l'urbain dense) et d'autres construites et mobilisées en tant qu'alternatives comme la banlieue (ex : Bevort et Rousseau, 2013) ou le périurbain (ex : Berque, 2015). 42 Introduction Du récit de fondation révélateur de fragments historiques d'un territoire, le mythe revêtirait donc une fonction de structuration sociale par l'identification de qualités dans la forme urbaine, créées ou valorisées par l'urbanisme (morphologiques, architecturales, mobilières). L'action urbanistique viendrait restaurer le lien entre le mythe et la réalité. Or, cette fonction suppose l'existence d'un décalage antérieur à l'action urbaniste entre le récit mythologique et la réalité structurée par ce dernier. Dès lors, le mythe se révèle souvent associé, dans les travaux d'urbanisme ou de géographie urbaine, à un récit « menteur » qui déforme ou simplifie les représentations de la réalité territoriale (Berque, 2015). En lieu et place d'une analyse d'un récit mythologique, la notion de mythe est souvent mobilisée en urbanisme de manière partielle et dans un but rhétorique par la formulation duale « mythe ou réalité » (ex : Glenn, 1986 ; Toinard, 1996 ; Chapain et Polèse, 2000 ; Jouve, 2006 ; Gallez, Guerrinha, Kaufmann, Maksim et Thébert, 2008). Si le mythe est critiqué par l'urbanisme pour son décalage avec le réel, il convient de relever que l urbanisme peut aussi être le producteur ou le véhicule de ces représentations erronées (Chalas, 1989), particulièrement dans le champ du transport (Offner, 1993 ; Gallez, Guerrinha, Kaufmann, Maksim et Thébert, 2008). Dès lors, « lutter » contre les mythes, les « détruire », échapper à leur « tentation », les « dénoncer », engageraient la responsabilité du chercheur travaillant sur l'urbanisme (Veyret-Verner, 1971, p. 7, p. 8, p. 10, p. 29). Ainsi, le mythe représente, dans les travaux scientifiques relatifs à l'urbanisme, un discours sur les phénomènes urbains qui serait soit illusoire et rendu obsolète par l'amélioration de la connaissance par ces travaux, soit mensonger et mobilisé à des fins d'instrumentalisation que dévoileraient ces travaux. Autrement dit, le mythe est réduit à des « idées périmées » ou des « promesses fallacieuses » (Veyret-Verner, 1971, p. 43 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique La mythanalyse est l'analyse des modalités de reproduction, ajustées à une société, d'une structure mythique traditionnelle. A l'inverse, la « mythocritique » porte sur le récit (formes narratives, figures de style) et son contexte d'écriture et / ou de narration, s'étendant ainsi « à un champ plus large, celui des pratiques sociales, des institutions, des monuments autant que des documents » (Durand, 1996, p. 205). Opérer une mythocritique, revient à tenter de reconstruire la structure mythique pour soulever le système d'oppositions propre aux constructions sociales, notamment entre la nature (ordre instinctif, individuel et nocturne) et la culture (ordre rationnel, communautaire et diurne). Cette structure mythique serait de nature symbolique selon Gilbert Durand (1964), prenant ainsi le contrepied de la méthode d'analyse de Claude Lévi-Strauss (1974) qui décompose le mythe en une somme d'objets anthropologiques (rituels religieux, peintures, sculptures, vêtements, cosmétiques) dissociables et codifiables dans un tableau 14. La mythocritique s'intéresse aux filiations et continuités spatio-temporelles des mythes, jusqu'à considérer certains récits mythologiques comme des réécritures modernes (avec leurs problématiques et leurs refoulés scientifiques) de caractéristiques structurales de mythe antique. Par exemple, cela revient à penser une filiation entre les récits de Prométhée et de Frankenstein (Marret, 2006), qui suivraient un même schème mythologique mais connaitraient des variantes d'écriture. Le mythe moderne « réaffirme la pertinence [d'un mythe antique] () en une période qui se penche, d' manière nouvelle, sur les conséquences de la domination du Logos » (ibid., p. 28). Le schème mythologique convoque un déroulement connu ; à cet égard, il peut se révéler rassurant et il suscite plus facilement une adhésion qui s'exprimera alors de manière tacite. En somme, c'est un mouvement d'analyse inverse entre la mythanalyse et la mythocritique : au contraire de la mythanalyse qui se réfère à la reconstitution d'un « mythe idéal » pour y inclure les domaines psychologique et sociologique propres à une société, la mythocritique vise plutôt à mettre en évidence les motifs mythiques fondateurs qui orientent nos affects et, par là, préfigurent nos actions. Au regard du rôle des affects dans les professionnalités, il nous a semblé plus pertinent d'inscrire notre recherche dans une perspective mythocritique. 4.2 Deuxième hypothèse : des crises écologiques révélatrices de la constellation mythologique de l'urbanisme Nous ne prétendons pas ici recenser les mythes antiques qui peuplent l'urbanisme (ex : Chalas, 1989 ; Genestier, 2001) ou des mythes modernes fondés sur des objets urbanistiques (ex : Offner, 1993 ; Boulba-Olga et Grossetti, 2018). Nous ne cherchons pas non plus à proposer une géofiction qui questionne le rôle de certains mythes dans la construction de théories géographiques et leurs mises à l'épreuve empirique (ex : Lecoquierre, 2008 et 2014 ; Desbois, Gervais-Lambony et Musset, 2016), ni à dévoiler des objets urbanistiques dont les effets annoncés relèveraient davantage de croyances que d'une administration de la preuve (ex : Berdoulay, Castro et Gomès, 2001 ; Bevort et Rousseau, 2013 ; Berque, 2015). Il s'agit 14 Dans la méthode de Claude Lévi-Strauss, les mythèmes sont numérotés et étudiés de manière systématique par un tableau croisant les relations entre les mythèmes (en colonne) avec l'ordre narratif du mythe (en ligne). 44 Introduction plutôt d'identifier des schèmes mythologiques qui sont mobilisés tacitement dans la justification de pratiques modernes. Nous posons comme deuxième hypothèse que les crises écologiques, en faisant resurgir des externalités modernes, mettraient à mal une constellation mythologique de l'urbanisme et la dévoileraient ce faisant. L'affaiblissement du sentiment de maîtrise La diversité des enjeux écologiques contribue à une perturbation visible, croissante et irréversible des conditions d'exercice des métiers : dérèglement climatique et événements de canicule, finitude des ressources particulièrement énergétiques, fragmentation et appauvrissement des éco ystèmes faunistiques et floristiques, pollutions de l'air et gaz à effet de serre, érosion des sols et montée des eaux, salinisation des sols et désertification Les savoirs des métiers rencontreraient des difficultés dans leurs reproductions, car leurs conditions de validité et de pertinence ne correspondraient plus aux situations urbaines soumises aux crises écologiques. Souvent posées comme un nouveau paradigme des métiers (durabilité urbaine) et une nouvelle gouvernance des territoires (ex : Agenda 21 locaux), les crises écologiques sont toutefois rarement prises en compte comme un facteur de questionnement des savoirs mobilisés dans les métiers (Faburel, 2015b). Pourtant, les effets territorialement différenciés des enjeux écologiques témoigneraient d'une vulnérabilité urbaine plus forte par rapport aux autres milieux (îlots de chaleur urbain, pollution atmosphérique) ; il est possible de se demander quel a été le rôle de l'urbanisme dans la fondation de cette vulnérabilité (concentration des activités, imperméabilisation des sols). Le mythe de Prométhée : la conquête de la nature par la rationalité instrumentale Le mythe de Prométhée est fondateur de l'entendement aménagiste. Il renvoie à l'arrachement de l'homme à la nature puis sa maîtrise par la rationalité instrumentale et technique. Cette dernière sécuriserait les effets de l'action aménagiste, ce qui permettrait une penser par modèles. La certitude en la rationalité instrumentale et technique est tellement forte qu'elle devient croyance, rendant impensable d'éventuels éffets non intentionnels de l'action. Le mythe de Babel : la quête d'unité perdue Le mythe de Babel est particulièrement présent dans les représentations territoriales des métiers. Il est associé à la quête d'une totalité perdue qui rassurerait par son unité. L'idée de totalité dans l'unité confère aux praticiens le rôle de détenteur de l'unique parole audible au milieu de modes d'expression variés comme l'architecture, le paysagisme, le génie civil Le mythe d'Orphée : la libération de l'ordinaire par la connaissance Le mythe d'Orphée suggère la libération aux contraintes ordinaires d'existence par la connaissance voire l'initiation aux mystères. C'est le mythe du savoir salvateur et différenciateur d'un groupe par la révélation des « fondations » ; il confère au praticien une autorité et une responsabilité. Bien que nous inscrivons cette recherche dans une perspective mythocritique, il nous est paru nécessaire de rappeler les principaux mythèmes des récits mythologiques étudiés. Nous avons donc dû opérer un choix parmi les différentes variantes. Concernant le mythe de Prométhée, nous avons choisi la version proposée par Platon dans Le Protagoras (1997), car Platon mobilise le mythe dans son argumentation lors des impasses de la construction dialect de la pensée, pour venir combler des lacunes dans la connaissance (Lacan, 2001, p. 145). Les crises écologiques créent une brèche dans la dialectique ambition moderne / schème mythologique de l'urbanisme. Contrairement aux deux autres mythes, le récit de Babel est d'origine religieuse ; nous lui avons préféré la version de la Bible de Jérusalem produite par l'Ecole biblique et archéologique française de Jérusalem à partir d'un travail de comparaison des différentes versions historiques et traductions linguistiques, se rapprochant d'une mythanalyse. Enfin, nous avons retenu le récit d'Orphée dans Les métamorphoses d'Ovide (2012) qui traite fondamentalement des « hybrides », mi-homme mi-animal, mi-homme mivégétal ; cette version nous a semblé intéressante pour étudier les partages modernes dont la déliquescence crée des hybrides. 4.3 Troisième hypothèse : les professionnalités comme subjectivation des savoirs pratiques Notre troisième hypothèse pose les professionnalités comme un levier de dépassement de la dialectique Modernité / Mythologie, structurante de la légitimité de l'urbanisme et fortement éprouvée par les crises écologiques. Les professionnalités constitueraient une épreuve d'autofondation subjective des savoirs et habiletés jugés pertinents pour les métiers. Elles opéreraient par la réflexivité (Schön, 1994), c'est-à-dire un retour continu des praticiens sur les formes et contenus de l'agir pour ajuster leurs capacités d'action. La professionnalité comme hybridation par l'action des vies sociales et professionnelles L'entrée par les professionnalités donne à voir « les formes d'hybridation, sociales, pratiques, intellectuelles, qui se nouent dans l'action même, notamment entre pratiques militantes et activité professionnelle des personnes qui s'engagent. » (Champy et Israël, 2009, p. 7). Il s'agit de trouver « un équilibre savant mais toujours précaire entre les dimensions institutionnelles, organisationnelles et les dimensions personnelles, subjectives, engagées sur les lieux où ils [les praticiens] travaillent. » (Laval, 2009). Or, en dehors du syndicalisme, l'engagement dans le cadre d'une activité rémunérée est très peu étudié par les recherches, notamment car il échappe aux cadres (souvent normatifs) d'analyse d'une activité de métier et il convie une « forme particulière » du politique (Champy et Israël, 2009, p. 7). Ainsi, ce sont à la fois les pratiques de métier et les modes d'engagement politiques qui s'hybrident, « pouvant se traduire à la fois par des engagements d'une nature particulière, dont l'expertise est un ressort, et des pratiques professionnelles modifiées, pour les rendre compatibles avec les idéaux revendiqués. » (ibid., p. 9). Le politique apparaît alors comme ce qui fait sens entre ces registres temporels distincts et fait évoluer les limites de la sectorisation des activités. Toutefois, si le terme de professionnalité suppose réflexivité et autonomie (de penser), il ne signifie pas des individus arrachés de toute appartenance. Il ne s'agit pas d'opposer un individu-praticien à une communauté professionnelle, mais plutôt de considérer que les savoirs mobilisés dans les professionnalités puisent dans des expériences qui débordent des seuls parcours biographiques jalonnés de grandes étapes (baccalauréat, formations, postes). A cet égard, il nous semble important de préciser que cette hybridation des pratiques avec des actions militantes se réalise en même temps qu'une évolution de l'engagement vers une « individuation », c'est-à-dire une construction et une reconnaissance de la singularité du praticien construite par sa trajectoire (Ion, 2012). Cette singularité prend le contrepied de deux figures de l'individu que l'on retrouve dans l'action publique : l'usager (individu anonyme) et le citoyen (individu universel) (Ion, 2010). Or, l'individuation a des répercussions sur des domaines très variés comme la famille et la religion, mais aussi dans des sujets proches de ce qui nous concerne, comme « dans les rapports à la chose publique où l'exigence de parler en nom propre se fait jour, et dans les organisations militantes où personnalisation et publicisation peuvent aller de pair quand la tradition établissait une stricte séparation du privé et du public. » (ibid. pp. 77 - 78). L'individuation nous conforte sur l'importance de la mise en cohérence de deux sphères que 'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique la Modernité tend à dissocier : la sphère professionnelle avec l'activité rémunérée mise au service d'un intérêt général pour l'urbanisme versus la vie sociale des praticiens dans lesquelles ils expriment leurs convictions et leurs désirs de transformation, les traduisant parfois dans des engagements (militants, associatifs, informels). La réalisation des professionnalités profite donc du défaut d'hermétisme de ce partage moderne des activités d'un individu pour conférer un sens convictionnel à la pratique de métier (Laval, 2009). Les limites hypothétiques de la professionnalité : dérégulation néolibérale, violence symbolique et adéquationnisme méthodologique formation / emploi La notion étant peu employée et à l'occasion réduite à un synonyme de compétence, il s'avère difficile de penser les critiques adressées à la professionnalité. Néanmoins, l'exercice nous semble intéressant car il nous e à expliciter certains points et de positionner la « professionnalité » par rapport à d'autres courants de pensées. Tout d'abord, nous relevons un piège en assimilant la valorisation de la professionnalité à une forme de dérégulation qui jouerait le jeu de l'idéologie néolibérale (Boltanski et Chiapello, 1999). Cette assimilation s'explique par une lecture substantive de la professionnalité (la confondant avec les compétences) qui ne prend pas en compte l'origine sociale du terme. En effet, la professionnalité ne cherche ni la conciliation des points de vue dans un « consensus » (Benasayag et Del Rey, 2012) mais la défense d'un commun, et la professionnalité n'est pas une « flexibilité » des pratiques ce qui témoignerait d'une « adaptation relative » à des contraintes et par là « naturalise[rait] » le contexte initial (Arnsperger, 2005). Notre première hypothèse inscrit l'urbanisme dans une dialectique entre, d'une part, une ambition de Modernité et, de l'autre, la mobilisation de schèmes de penser mythologiques. De par son ancrage urbain (Castells, 1972), l'urbanisme réduit le mythe à un récit illusoire voire trompeur sur une forme urbaine et sa qualité sociale, qu'il chercherait à redresser en se posant comme un savoir scientifique répondant aux critères positivistes de neutralité et de distanciation (hypothèse 1). Néanmoins, les crises écologiques perturberaient cet équilibre par la résurgence de certains refoulés (hypothèse 2). Ceux-ci relèveraient d'arbitrages politiques sous-jacents à la production et la mobilisation des pratiques de l'urbanisme, concernant surtout les instruments de l'urbanisme (mythe de Prométhée), les représentations territoriales de l'urbanité (mythe de Babel) et la définition créatrice de la posture d'expert (mythe d'Orphée). Ces discontinuités révélées dans la justification rationnelle de l'urbanisme pourraient être un espace d'engagement des praticiens pour la construction de sens de leurs métiers par une diversification et une hybridation des savoirs embarqués (hypothèse 3). En conviant des savoirs issus de l'engagement, en faisant preuve de réflexivité, en inscrivant leurs pratiques dans un désir de ville, en faisant valoir leurs tâtonnements, etc., les praticiens pourraient façonner une subjectivation des savoirs, c'est-à-dire une construction politisée et située des connaissances par la trajectoire du praticien, le contexte d'emploi (poste, structure), la situation (territoriale, partenariale, etc.) de professionnalité 5. Corpus de recherche : démarche, publics, matériaux Notre corpus de recherche a été construit pour saisir les professionnalités de l'urbain qui, en tant que savoirs cachés de l'agir professionnel » (Schön, 1994), demandent un cadre interprétatif différent de celui des savoirs formels. Plus précisément, à l'aune de la nature revendicative des professionnalités, nous avons privilégié une démarche compréhensive de recueil des informations, permettant de reconnaitre la portée créatrice des praticiens dans le renouvellement de leurs métiers. Egalement, à l'aune de la subjectivité des choix animant les professionnalités, nous avons puisé nos matériaux dans des situations d'énonciation variées. 49 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique 5.1 Une démarche compréhensive : dévoiler la capacité créatrice des praticiens par une diversité de discours et de matériaux Si un décalage entre les discours praticiens et la prescription urbanistique des pratiques a pu émerger, c'est en raison de la démarche de recherche dite compréhensive que nous avons mobilisée. En effet, celle-ci laisse les praticiens identifier les enjeux et les problématiques, en choisissant des termes porteurs de sens selon eux et auxquels ils apportent leurs propres significations. Théorisée notamment par Marie-Noëlle Schurmans (2003), la démarche compréhensive « inverse le mode de construction de l'objet » (Charmillot et Dayer, 2007, p.136), c'est-à-dire essaie d'éviter une construction a priori des objets de recherche par des catégories d'analyse, des méthodes d'objectivation et des instruments de recueil de données. Elle suppose ainsi la préexistence des enjeux et sujets de recherche, par exemple sous la forme d'une « lacune de connaissance » que la recherche essaierait de combler. Cette démarche compréhensive s'est traduite, dans notre corpus, par une prévalence accordée aux discours praticiens pour caractériser les évolutions de leurs métiers et de leurs pratiques. Nous avons favorisé des méthodes qui mettent en discours les savoirs pratiques afin d'en dégager le sens que les praticiens leurs confèrent. Nous considérons, en effet, que la mise en mots oblige les praticiens à interrompre leurs activités et formaliser leurs questionnements. Plus qu'une approche descriptive ou informative du langage, nous lui assignons une fonction de mise en intelligibilité du « monde » du narrateur (Berger et Luckmann, 1997). Concrètement, nous nous sommes intéressés à l'omniprésence ou l'absence d'un terme, sa polysémie ou sa signification plutôt partagée par les praticiens, l'incapacité à exemplifier une idée ou les liens qui sont dessinés avec d'autres phénomènes. Une diversité de matériaux pour une diversité de discours Cette thèse s'appuie sur un corpus composé de différents matériaux de recherche, reposant pour l'essentiel sur les méthodes d'entretiens semi-directif et de séminaire d'échanges, complétées d'une méthode de recensement et d'analyse de discours formalisés par l'écrit, ainsi que de temps d'échanges informels avec des praticiens et des étudiants. A cet égard, cette recherche croise des discours aussi bien individuels que collectifs, formalisés dans des écrits que de production progressive lors d'échanges collectifs, institués par le contexte d'énonciation que libérés par des entretiens anonymisés. Ces matériaux ont été composés dans des conditions différentes afin de faire émerger des professionnalités qui peuvent éclore de situations pratiques extrêmement multiples (Laval, 2009). D'une part, certains s'inscrivent dans des recherches plus vastes et ont donné lieu à des contrats de recherche (de un à trois mois) avec des structures publiques : un organisme de recherche public (Laboratoire d'Excellence Intelligences des Mondes Urbains, dit LabEx IMU), une administration centrale de l'Etat (Commissariat Général à l'Egalité des Territoires, dit CGET) et un service de la recherche (Ministère de l'Environnement, de l'Energie et de la Mer). A cet égard, ces matériaux ont fait l'objet de rapports de recherche indépendants de ce manuscrit et dont les anlayses sont ciblées en fonction des recherches plus vastes dans lesquelles ils s'inscrivent. Cette thèse ne reprend pas strictement les analyses de ces rapports, mais mobilise certains de leurs matériaux - dans le respect de leurs contextes de réalisation qui nous permettent de renseigner les professionnalités de l'urbain. Ces matériaux issus d'une recherche plus vaste sont : l'enquête auprès de praticiens (Girault, 2015), l'observation participante à la préfiguration d'un programme de recherche (Girault, 2016a), et le recensement et l'analyse d'études territoriales expérimentales (Faburel et Girault, 2017a ; Girault, 2017a). Enfin, les autres matériaux, soit l'enquête auprès d'étudiants et le recensement analytique des discours des associations dites professionnelles, ont été réalisés dans la seule perspective de nos recherches et ont donné lieu à des valorisations sous la forme d'articles scientifiques (Girault, 2016b, 2018a et 2018b) et d'échanges au sein du Réseau des Activités des Métiers de l'Architecture et de l' isme (RAMAU - Girault, 2018c). Méthodes non-directives : entretiens semi-directifs et séminaires d'échanges L'identification des doutes et questions praticiens sans les restreindre à des problématiques scientifiques prédéfinies suppose un principe de non-directivité qui « impose au chercheur d'introduire une question de départ puis de se cantonner à des relances-miroir, reprenant strictement les mots et expressions de l'enquêté. » (Demazière, 2007a, p. 90). Or, la nondirectivité « est devenue progressivement, dans les sciences sociales, une espèce de norme académique, enseignée comme une méthode canonique. » (ibid.). A cet égard, il a été aisé de 51 Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique renseigner des méthodes non-directives de recueil d'informations et nous avons eu recours à deux d'entre elles : l'entretien semi-directif et le séminaire d'échanges. Un entretien semi-directif est « un entretien au cours duquel un « chercheur » (lequel peut être un étudiant en tant que jeune chercheur) demande à une personne, que nous désignerons tout au long de ce texte comme « sujet », de lui raconter tout ou partie de son expérience vécue. » (Bertaux, 2016, p. 11). Cette méthode a donne lieu à différentes variantes en termes de conduites d'entretien selon la nature différenciée de leurs sujets, qu'il s'agisse d'un entretien compréhensif (Kaufmann, 1996) ou approfondi (Demazière et Dubar, 2004), ethnographique (Beaud, 1996) ou socioethnographique (Bertaux, 2016), biographique (Pruvost, 2011) Largement répandue, cette méthode a été rapidement intégrée dans cette recherche dès l'enquête menée auprès de praticiens (en 2015) et fréquemment sollicitée auprès d'un public varié (praticiens, chercheurs et étudiants), jusqu'à constituer une part importante de notre corpus (40 entretiens en total). A l'inverse de cette rapidité de tenue des entretiens semi-directs, la méthode des séminaires d'échanges a demandé davantage de temps de préparation et, surtout, un appui de la part de notre directeur de recherche pour nous familiariser avec la méthode et nous aider dans l'animation qui recquiert la présence d'au moins deux personnes (une en charge de poser les questions prédéfinies, une seconde davantage observatrice du tour-de-parole et de la circulation des arguments). Nous avons ainsi pu observer à deux reprises cette méthode lors d'autres recherches (Faburel, 2014a et 2015a ; Faburel et Girault, 2015a) 15 avant de la mettre en place à deux reprises (cf. enquête auprès de praticiens et analyse d'études territoriales expérimentales). Si la tenue de sémin d'échanges suppose certes des méthodes d'animation pour garantir le tour-de-parole, la dificulté principale réside, selon nous, dans la préparation concomitante du tour-de-table et des questionnements de recherche. En effet, ces séminaires étaient l'occasion 1/ d'approfondir certains sujets traités en entretien, 2/ de confronter des situations de mésentente observées c'est-à-dire des situations où les praticiens emploient le même terme mais selon des conditions et des finalités distinctes (ex : le terme d' « atelier »), 3/ de construire une compréhension commune des enjeux liés aux mutations urbaines et crises écologiques permettant 4/ d'envisager une vision commune des évolutions espérées des pratiques et des stratégies pour y parvenir (rapports aux institutions, détournements méthodologiques). Pour ce faire, cette méthode demande un temps long de préparation soit par l'analyse de travaux menés par les futurs participants au séminaire (cf. le séminaire sur les études territoriales expérimentales), soit par un temps d'entretien préalable (cf. l'enquête auprès de praticiens). Cela permet de situer les enjeux identifiés par les praticiens et de souligner des questionnements qui leur seraient communs. Méthodes de tenue et de référencement des matériaux : assurer une visibilité Au regard des diverses conditions de production de nos matériaux, il a été important pour nous d'assurer une systématique dans l'analyse et la tenue de chaque méthode. Ainsi, au regard de l'importance conférée au choix des mots, les entretiens et les séminaires ont été systématiquement - sous autorisation - enregistrés 17 et retranscrits de manière anonyme. Ainsi, malgré la diversité des publics rencontrés (praticiens, enseignants-chercheurs et étudiants) et des trois conditions de recherche (préfiguration d'une formation de master, observation participante de la préfiguration d'un programme de recherche, travaux de recherche personnels), nous avons toujours suivi la même méthode pour la tenue des 41 entretiens semi-directifs. La prise de contact s'est faite essentiellement par courriel avec des indications concernant le sujet de thèse et éventuellement le contexte de la recherche (ex : préfiguration d'une formation de master). Les mots ont été choisis pour rester le plus large possible, par exemple nous annoncions vouloir parler de « l'évolution de [leur] métier et des savoirs rendus nécessaires en conséquence », sans préciser la nature de cette évolution (contrainte, forcée, volontaire) ni ses causes (institutionnelle, écologique, économique), sans spécifier si les savoirs rendus nécessaires étaient manquants ou d'ores et déjà embarqués dans la pratique. Les entretiens ont réalisés en privilégiant l'échange de visu (à l'exception de six d'entre eux, dont cinq réalisés par téléphone et un par vidéoconférence), soit dans le bureau de la personne pour l'essentiel, dans un espace neutre quelque fois (ex : café) ou dans un bureau à l'université pour l'essentiel des entretiens menés avec des étudiants. Toutes les personnes rencontrées se sont prêtées volontiers à la discussion, même lorsque les questions pouvaient revêtir une dimension plus personnelle (ex : présentation de sa trajectoire avec ses bifurcations, explicitation de ses choix de formation ou de poste) ; cela a donné lieu à des entretiens approfondis de 1h15 en moyenne. 17 Seuls deux entretiens n'ont pas été enregistrés sur les 40 réalisés. Professionnalités de l'urbain et crises écologiques. Politiser l'urbanisme et ses métiers par la reconnaissance de leur constellation mythologique Nos deux séminaires ont été réalisés avec une vingtaine de participants à chaque fois pour faciliter la circulation de la parole, réalisés selon deux modalités temporelles différentes : soit un séminaire resserré sur une journée (cf. l'analyse des études territoriales expérimentales) 18, soit un séminaire composé de trois séquences courtes (de trois heures) renouvelées tous les un ou deux mois pendant cinq mois (cf. enquête praticiennne) 19 et faisant l'objet à chaque fois d'un compte-rendu transmis aux participants. Si nous avons mis en place une systématique dans la réalisation et l'analyse des différents matériaux, nous avons fait le choix de ne pas en rechercher pour le référencement dans ce manuscrit. En effet, chaque matériau étant mobilisé de manière resserrée dans une partie ou une sous-partie, une indication initiale (dans le corps du texte ou en note de bas) suffit pour identifier les conditions de réalisation du corpus mobilisé, ce qui est plus facile à la lecture qu'un système codifié à retenir. De plus, cela nous permet de jouer des résultats dans l'argumentation de notre propos. Par exemple, pour l'enquête auprès de praticiens, nous avons fait le choix de restituer ces matériaux sans préciser la fonction et la structure d'appartenance du praticien afin de souligner la compréhension commune des praticiens rencontrés quant à l'évolution de leurs métiers ; à l'inverse, les entretiens étudiants ont fait l'objet d'une restitution classique avec le choix d'un prénom de consonnance (sociale, territoriale) similaire la précision de l'âge et de l'année de formation, afin de pouvoir reproduire les trajectoires dans leurs continuités, ruptures, bifurcations Nous présentons ci-dessous chaque matériau en les différenciant selon leurs conditions de production. Nous mettons en annexes (cf.	9,053
20/hal.archives-ouvertes.fr-hal-00541384-document.txt_1	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	2,670	4,213	Le problème inverse acoustique vu comme la solution d'un problème de Cauchy associé à l'équation de Helmholtz Lang Le problème inverse acoustique vu comme la solution d'un problème de Cauchy associé à l'équation de Helmholtz. Christophe Langrenne, Alexandre Garcia Conservatoire National des Arts et Métiers, 292 rue Saint Martin, 75141 Paris cedex 3, {christophe.langrenne,alexandre.garcia}@cnam.fr Depuis maintenant trente ans, l'imagerie acoustique a développé de nombreuses méthodes visant à caractériser des sources sonores : holographie de champ proche (NAH) en géométries simples, méthode d'élements finis de frontières (BEM) pour des géométries plus complexes pour ne citer que les plus connues. Ces méthodes permettent de calculer l'opérateur de propagation entre les grandeurs de la structure à caractériser et les grandeurs mesurées. L'opération de rétro-propagation amplifie fortement le bruit de mesure car ce problème inverse est "mal-posé" au sens de Hadamard. Des méthodes de régularisation sont donc nécessaires pour stabiliser la résolution et trouver des solutions réalistes. Généralement, par souci d'économie, seule la pression est mesurée autour de l'objet, mais l'essor des frontaux multivoies permet l'acquisition d'un nombre toujours plus grand de données. Ainsi, il est commun de mesurer la pression et son gradient à l'aide de sondes microphoniques calibrées ou des sondes pression vitesse, sur un maillage conséquent. Le problème inverse acoustique peut alors être vu comme la solution d'un problème de Cauchy associé à l'équation de Helmholtz : soit un domaine borné, connaissant les données de pression et de vitesse sur une partie des limites du domaine, on cherche à les déterminer sur la partie restante. L'idée de base est de transformer ce problème mal-posé en deux problèmes bien posés, et de faire coı̈ncider les deux solutions. Différentes méthodes mathématiques ont été développées, mais ne semblent pas avoir été utilisées en acoustique. Nous présenterons deux de ces méthodes (Kozlov, opérateur de Steklov-Poincaré) et les illustrerons sur des simulations numériques. 1 Introduction La caractérisation de sources acoustiques est un sujet important pour les industriels car à partir de mesures, on est à même de localiser les fortes vibrations et les zones responsables du rayonnement de l'objet ou de simuler a posteriori la présence de la source dans d'autres milieux. Si des mesures directes existent, de type accélérométriques ou par vibrométrie laser, elles sont, pour la première lourde à mettre en oeuvre, et pour la seconde, couteuse. Bons nombres d'études visent alors à résoudre le problème inverse acoustique qui consiste à partir de données de pressions en champ proche de reconstruire la vibration de la structure. Récemment, S.F. Wu [1] propose une synthèse des méthodes utilisées en acoustique. De nos jours, la puissance de calcul toujours plus importante des ordinateurs permet des modélisations numériques de plus en plus fine. Ainsi, les méthodes d'éléments finis de frontières permettent des traitements d'objets plus étendus et pour des fréquences plus élévées. Apparaissent en parallèle des systèmes d'acquisition multivoies comportant des centaines de canaux de mesures et des capteurs permettant l'acquisition à la fois de la pression et de son gradient. On peut alors envisager la résolution du problème inverse comme la solution du problème dit de Cauchy. Considérant un domaine borné, les conditions aux limites dans ce problème, sont sur-abondantes (pres- Fig. 1: Configuration du problème sion et son gradient) sur une partie du domaine, tandis que sur une autre, elles sont inexistantes. Ce genre de problème est courant dans de nombreux domaines de la physique, lorsqu'une partie du domaine est inaccessible à la mesure (thermique, mécanique, opto-électronique, etc). Mathématiquement dans notre cas, le problème de Cauchy s'écrit sous la forme suivante :  dans Ω, (4 + k 2 )p = 0 p = P m, ∂n p = V m sur Γc, (1) p = ps, ∂n p = ∂n ps sur Γi, où P m et V m sont les grandeurs connues sur la limite du domaine Γc et les grandeurs ps et ∂n ps sont les inconnues à déterminer sur la frontière Γi (voir figure 1). Pour résoudre ce problème (1), une première solution consiste à écrire la formulation intégrale sur les limites du domaine et à ré-organiser les opérateurs, correspondant aux données connues et aux inconnues. Le système discrétisé obtenu est alors un système linéaire classique où l'on doit inverser un opérateur. 2 Formulation intégrale frontière sur le domaine de La formulation intégrale correspondant au système (1) est la suivante : ZZ P m ∂n G(s, s0 ) − V m G(s, s0 ) dS Γc ZZ ps ∂n G(s, s0 ) − ∂n ps G(s, s0 ) dS = 0 . (2) + Γ i Après discrétisation à l'aide d'une formulation isoparamétrique [2], on obtient le système matriciel : Dc P m − Mc V m + Di ps − Mi ∂n ps = 0, et après ré-organisation des grandeurs connues et inconnues : ps Pm Di −Mi = −Dc Mc, (3) ∂n ps Vm soit, une formulation classique de la forme Ax = b, où l'opérateur A est mal-conditionné. Sans précautions, des erreurs sur le second membre vont alors être démesurément amplifiées, et la solution obtenue sera irréaliste. Le problème rencontré est identique à celui de la résolution conventionnelle, où A représente l'opérateur de propagation entre la distribution de vitesse sur la structure de l'objet et celle de pression mesurée en champ proche. Le caractère évanescent du champ proche impose une limite, liée au rapport signal à bruit des mesures. Après une décomposition en valeurs singulières de l'opérateur A, la technique de régularisation consiste à filtrer les termes évanescents correspondants à des valeurs singulières trop petites. Lorsque le rapport signal à bruit est connu, nous pouvons utiliser le principe de Morozov basé sur une information a priori de la qualité des mesures. Sans connaissance, a priori, de ce rapport, d'autres techniques ont été développées dont la plus utilisée est la "L-curve", qui permet de faire un compromis entre la norme de la solution et la norme des erreurs de propagation. Effectivement, plus le filtrage est fort, plus la norme de la solution diminue, alors que l'erreur de propagation augmente [3]. 3 Séparation en deux problèmes bien posés sous- Le système (1) peut se décomposer en deux soussystèmes bien posés, de la manière suivante :  (4 + k 2 )u1 = 0 dans Ω, u1 = P m sur Γc, (4) ∂n u1 = η sur Γi, et  (4 + k 2 )u2 = 0 dans Ω, ∂n u2 = V m sur Γc, u2 = τ sur Γi. (5) Ici, le système (4) correspond au problème aux limites de Dirichlet et Neumann, tandis que l'autre système (5) correspond au problème aux limites de Neumann et Dirichlet. Il existe des variantes à ces systèmes en combinant d'autres conditions aux limites de type Robin. La solution du système (1) est alors la solution des systèmes (4) et (5), lorsque celles-ci coı̈ncident : u1 = u2 = u. Pour ce faire, Kozlov et al [4] utilisent un algorithme itératif qui résoud alternativement les systèmes (4) et (5). Une autre solution consiste à calculer l'opérateur de Steklov-Poincaré agissant sur la frontière Γi, technique utilisée lorsque l'on décompose le domaine en plusieurs sous-domaines et que les inconnues se trouvent sur une frontière commune aux différents sous-domaines [5]. Les paragraphes suivants décrivent la mise en oeuvre de ces deux techniques au problème d'imagerie. 3.1 Algorithme de Kolzov Kozlov et al [4] propose un algorithme itératif basé sur la résolution des systèmes (4) et (5), alternativement. L'auteur montre que cet algorithme converge vers la solution pour un opérateur elliptique. On choisit pour initier l'itération une solution η 0 et l'on résoud le système (4). Cette solution permet de déterminer la condition limite τ 0 qui sera utilisée dans le problème (5), et ainsi de suite. Malheureusement, dans nos essais, et comme le fait remarquer Marin [6], cette algorithme ne converge pas toujours pour l'équation de Helmholtz. Aussi, récemment, Johansson et Kozlov [7] proposent une variante de leur méthode adaptée à l'équation de Helmholtz. Cependant, la mise en oeuvre de cette variante pose le problème de l'arrêt du processus itératif. Une régularisation est nécessaire pour obtenir une solution réaliste, et le nombre d'itérations joue ce rôle. Les auteurs supposent connue la qualité des mesures et ils peuvent ainsi utiliser un critère d'optimalité de déviation donné par Morozov [8]. 3.2 Opérateur de Steklov-Poincaré Pour calculer l'opérateur de Steklov-Poincaré [5], les solutions de (4) et (5) se décomposent comme la somme de deux fonctions u1 = ω10 +ω1∗ et u2 = ω20 +ω2∗, solutions des systèmes suivants :  (4 + k 2 )ω10 = 0 ω0 = 0 10 ω1 = λ  (4 + k 2 )ω1∗ = 0 ω∗ = P m 1∗ ω1 = 0  (4 + k 2 )ω20 = 0 ∂n ω 0 = 0 0 2 ω2 = λ  (4 + k 2 )ω1∗ = 0 ∂n ω ∗ = V m ∗ 2 ω2 = 0 dans Ω, sur Γc, sur Γi, (6) dans Ω, sur Γc, sur Γi, (7) dans Ω, sur Γc, sur Γi, (8) dans Ω, sur Γc, sur Γi, (9) où ω10 et ω20 sont les extensions harmoniques dans Ω de λ sur Γi, notée respectivement H− (λ) et H+ (λ). Tandis que ω1∗ et ω2∗ sont les extensions harmoniques dans Ω de P m et V m de Γc, notée R− (P m ) et R+ (V m ). Pour que les deux solutions coı̈ncident, on utilise la condition sur Γi, du type : ∂u2 (λ) ∂u1 (λ) =, ∂n ∂n qui se traduit par le système à résoudre en λ : Sλ = χ sur Γi, (10) (11) où χ := −∂n R− (P m ) + ∂n R+ (V m ) (12) et S est l'opérateur de Steklov-Poincaré, défini par : S := ∂n H− (λ ) − ∂n H+ ( λ). (13) Le système (11) se révèle également "mal-posé" et nécessite une régularisation. 4.1 Simulations numériques Configuration des tests Pour les simulations numériques la configuration des tests est la suivante. Les mesures sont simulées sur une sphère de rayon r1 = 0.3 m. Les données reconstruites se situent sur une sphère de rayon r2 = 0.2 m. Le maillage des deux surfaces est identique et est constitué de 24 octangles à 8 noeuds, soit 72 points pour chaque surface (figure 2). Les sources testées sont des sources simples dont on connaı̂t une expression analytique pour la pression et son gradient. Nous avons pris deux sources tests à l'intérieur de la surface de rayon r2 : un dipôle centré et un quadripôle latéral centré. La fréquence d'étude choisie arbitrairement à 150 Hz, relativement basse car notre discrétisation est grossière et n'autorise pas une étude à des fréquences élevées. Fig. 2: Maillage d'une sphère en 24 octangles. 4.2 Comme les systèmes sont résolus sous forme discrète, les opérateurs issus des formulations intégrales ne sont qu'une approximation des opérateurs continus. Ils sont obtenus à l'aide d'une formulation iso-paramétrique décrite par Seybert et al [2] et basée sur des fonctions de forme d'ordre 2, adaptées à des octangles à 8 noeuds. Dans ces simulations, nous avons perturbé les données avec un taux de 1% d'erreur, ce qui correspond à des erreurs sur les pressions de l'ordre de ±0.2 dB, représentatives d'erreurs de mesures. 4.3 Méthode de régularisation Nous avons vu que pour obtenir des solutions réalistes, une méthode de régularisation est nécessaire. Nous utilisons celle de Tikhonov qui reste largement employée pour tout problème dit "mal-posé" [9]. Elle est basée sur la minimisation de la fonctionnelle suivante : Φα (xα ) = kAxα − bk2 + αkLxα k2, (14) où A est l'opérateur à inverser obtenu par les différentes méthodes, α le paramètre de régularisation et L un opérateur de régularisation. Généralement, l'opérateur de régularisation est pris sous la forme d'une matrice identité pour minimiser la norme de la solution. Effectivement, l'opérateur A est mal-conditionné, ce qui provoque de fortes oscillations et des amplitudes totalement irréalistes, le second membre de (14) joue le rôle de stabilisateur de la solution. Avec L = Id, des oscillations subsistent (voir figure 3), surtout sur la grandeur du gradient de pression. Nous utilisons alors un opérateur lissant qui correspond à minimiser la dérivée d'ordre 1 de la solution. Cet opérateur est approximé par différence finie. Enfin, la résolution dépend du paramètre de régularisation α. Si celui-ci est trop élevé, la solution sera atténuée. Cela permet d'avoir une idée de la solution, mais sans les détails. On filtre alors excessivement les termes évanescents pour ne garder que les termes propagatifs. Si par contre, ce paramètre est trop faible, des oscillations apparaissent et la solution est trop sensible aux erreurs de mesures. Il faut donc faire un compromis et différentes techniques ont été développées pour trouver celui-ci. Comme nous l'avons vu, le paramètre de régularisation est fortement dépendant du rapport signal à bruit de mesure. Si l'information sur l'erreur des mesures est connue, on peut utiliser le principe d'optimalité de déviation donné par Morozov. Une application est donnée dans [9]. Sans cette information, la "L-curve" permet de faire un compromis entre la norme de l'erreur de propagation, premier terme terme de (14) et la norme de la solution, second terme de (14). Dans certain cas, la courbe ne possède pas de cette caractéristique de "L" et le critère optimal ne peut être déterminé. Nous utilisons ici un critère quasi-optimal donné par Morozov [8] : dxα k2, (15) α dα qui est interprété comme la sensibilité de la solution régularisée par rapport aux variations du paramètre α. Fig. 4: Effet de l'opérateur lissant d'ordre 1 sur l'erreur relative correspondant au cas du dipôle centré. que la résolution du problème de Cauchy améliore sensiblement les résultats par rapport à ceux d'une rétropropagation classique. Les deux formulations (3) et (11) donnent des résultats quasiment identiques. Sur chacune des quatre courbes, un cercle permet de localiser la valeur du paramètre de régularisation donné par la formulation (15), qui ne correspond pas forcément au paramètre optimal. Sans information sur la qualité des mesures, ce critère reste cependant très performant, associé à la rétropropagation des gradients, la formulation intégrale de Cauchy et l'opérateur de Steklov-Poincaré. Par contre, il l'est moins pour la rétropropagation des pressions. Fig. 5: Erreurs relatives dans le cas du dipôle centré pour les différents opérateurs. 5 Fig. 3: Effet de l'opérateur lissant d'ordre 1 sur le gradient d'un dipôle centré. Les figures 5 et 6 montrent les erreurs relatives correspondant aux cas du dipôle et du quadripôle centrés. Nous pouvons remarquer que la rétropropagation à partir des gradients de pression donne de meilleurs résultats que ceux obtenus à l'aide des pressions. Nous voyons Conclusion Le développement technologique (acquisition multivoies, puissance des ordinateurs) permet d'envisager la résolution du problème inverse acoustique comme la solution d'un problème de Cauchy de l'équation de Helmholtz. A partir de données surabondantes sur une partie des frontières du domaine (pressions et gradients), les inconnues sur une autre partie peuvent en être déduites. En ce qui concerne la rétropropagation classique de données, deux formulations peuvent être obtenues : soit [7] B. T. Johansson and V. A. Kozlov, "An alternating method for Cauchy problems for Helmholtztype operators in non-homogeneous medium", IMA Journal of Applied Mathematics 74, 62-73 (2009). [8] V. Badeva, V. Morozov, "Problèmes incorrectement posés, théorie et applications", éd. Masson, 1991. [9] E. G. Williams, "Regularization methods for nearfield acoustical holography", J. Acoust. Soc. Am. 110 (4), 1976-1988 (2001). Fig. 6: Erreurs relatives dans le cas du quadripôle latéral centré pour les différents opérateurs. à partir des pressions, soit à partir des gradients mesurés. Nous avon s vu que celle qui est associée aux gradients donne de meilleurs résultats. Les formulations associées au problème de Cauchy améliorent encore sensiblement les solutions dans les simulations ici traitées. Cependant une régularisation de Tikhonov est nécessaire pour obtenir des solutions réaliste. Un opérateur lissant d'ordre 1, construit par différence finie, améliore grandement les résultats. Le paramètre de régularisation donné par un ère de quasi-optimalité se révèle très performant. Ces résultats préliminaires permettent d'envisager de manière optimiste ces méthodes sur des cas expérimentaux..	5,743
66/tel.archives-ouvertes.fr-tel-02000942-document.txt_3	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	7,689	12,895	49 Chapitre I – Les généralités 50 Chapitre II Méthodes et techniques expérimentales utilisées Chapitre II – Méthodes et techniques expérimentales utilisées 52 Chapitre II – Méthodes et techniques expérimentales utilisées Chapitre II – Méthodes et techniques expérimentales utilisées I. Introduction Les principales méthodes et techniques expérimentales utilisées au cours de ce travail de thèse sont décrites dans ce chapitre. Tout d'abord sont décrites les méthodes de caractérisation telles que l'analyse ICP-OES qui permet d'identifier et de quantifier les métaux de transition (manganèse, cobalt et nickel) et le lithium présents dans le matériau, l'analyse ATG qui permet de mesurer une variation de masse au cours d'un traitement thermique et l'analyse DSC qui permet de déterminer la stabilité thermique des matériaux à l'état chargé de la batterie et d'identifier la nature des flux de chaleur. Dans un deuxième temps est décrite la diffraction des rayons X sur poudre qui permet de caractériser la structure cristalline des matériaux. La troisième partie est dédiée à la comparaison des techniques d'analyses élémentaires par microscopie (MEB-EDX, EPMA (microsonde) et STEM-EDX) utilisées pour caractériser les gradients de concentrations (étendue et composition) ainsi qu'à celle des méthodes de préparation des échantillons (la méthode cross section polisher pour l'identification au MEB-EDX et EPMA et la méthode FIB pour l'identification au STEM). La dernière partie est consacrée à expliquer le protocole de préparation des accumulateurs afin de les tester électrochimiquement. 53 Chapitre II – Méthodes et techniques expérimentales utilisées II. Méthodes de caractérisation de la composition, de la structure et de la stabilité thermique des matériaux II. 1. Dosages chimiques du lithium et des métaux de transition par ICP-OES (spectrométrie de masse à plasma induit – spectrométrie à émission optique) La spectrométrie d'émission optique à plasma à couplage inductif est une méthode d'analyse chimique permettant de mesurer quantitativement la teneur de la quasi-totalité des éléments simultanément contenus dans un matériau. Les seuls éléments qui ne peuvent pas être dosés par la méthode sont le carbone, l'hydrogène, l'oxygène, l'azote, l'hélium et les halogènes. Dans le cas de nos matériaux, les éléments dosés sont le lithium et les métaux de transition tels que le manganèse, le nickel et le cobalt. Son nom usuel est « ICP - OES », il est issu de la dénomination anglaise « Inductively Coupled Plasma - Optical Emission Spectrometer ». L'appareil utilisé est un spectromètre Varian 720-ES. J'ai réalisé ces analyses en collaboration avec Laetitia Etienne du service collectif d'Analyses Chimiques et Thermiques de l'ICMCB. Un spectromètre ICP peut être divisé en quatre parties [1]-[2] comme illustré sur la figure 24 : - Le système d'introduction de l'échantillon - Le système d'atomisation et d'excitation - - Une étape de préparation des échantillons est nécessaire avant son analyse, j'ai donc réalisé toutes les mises en solution ainsi que les préparations d'échantillons : environ 10 mg de matériau est dissout à chaud dans 5 ml d'une solution d'acide chlorhydrique concentrée (37%) puis dans 5 autres ml d'une solution d'acide nitrique, puis é en complétant une fiole jaugée à 100 ml avec de l'eau distillée ultra pure. Un blanc d'analyse est réalisé pour retirer le bruit de fond correspondant à l'acide utilisé pour acidifier les échantillons ainsi que pour l'eau utilisée pour la dilution. Un passeur automatique d'échantillons couplé à une pompe péristaltique introduit la solution précédemment préparée dans une chambre de nébulisation où elle est transformée en un aérosol liquide composé de micro-gouttelettes à l'aide d'argon gazeux. Deux mises en solution ont été réalisées pour chaque échantillon et trois mesures ont été faites pour chacune d'entre elles, afin de vérifier la reproductibilité et la précision des mesures. 55 Chapitre II – Méthodes et techniques expérimentales utilisées Figure 24 : Schéma de l'appareillage d'un spectromètre ICP -OES II. 2. Analyse thermogravimétrique L'analyse thermogravimétrique (ATG) est une technique d'analyse qui permet de déterminer la variation de masse d'un échantillon au cours d'un traitement thermique T = f(t) (T température et t temps) sous atmosphère contrôlée. L'augmentation ou la diminution de la température engendre des transformations chimiques ou physiques (décomposition, évaporation, oxydation, réduction,) qui s'accompagnent d'une perte ou d'un gain de masse. Le dispositif, comme illustré sur la figure 25, est constitué d'une microbalance et d'un système de chauffage, l'échantillon est donc pesé et la ligne de base établie. En l'absence de réaction ou de transformation, la variation de masse sera faible. Lorsqu'une transformation de l'échantillon intervient, des réactions chimiques peuvent libérer ou former des espèces qui entrainent une variation plus ou moins importante de la masse. Ces analyses ont été réalisées par Philippe Dagault au sein du groupe de recherche de l'ICMCB en utilisant un appareil commercial TA instruments Q600. Elles ont permis de déterminer la température de décomposition des carbonates de métaux de transition MCO3. Dans ce cas, les expériences ont été réalisées sous air, avec une montée en température de 2°C/min jusqu'à 900°C, un palier de 7h à 900°C et une descente programmée à 20°C/min. 56 Chapitre II – Méthodes et techniques expérimentales utilisées Figure 25 : Représentation schématique d'une ATG II. 3. Mesure de la stabilité thermique par calorimétrie différentielle à balayage Afin d'étudier la stabilité thermique des matériaux du type Li1+xM1-xO2 à l'état chargé de la batterie Li-ion (c'est-à-dire lorsqu'ils sont désintercalés en lithium et donc oxydés), l'analyse thermique DSC (pour Differential Scanning Calorimetry) est utilisée. Les batteries Li//Li1+xM1xO2 sont chargées jusqu'à un potentiel de 4,8 V vs Li+/Li, après un premier cycle de formation à régime C/10 entre 2,5 et 4,8 V vs Li+/Li. A l'issue de ce cyclage, un fragment d'électrode non lavée de 5 mg est prélevé et immédiatement scellé, en boîte sèche, dans un creuset « haute pression ». J'ai réalisé la préparation des échantillons. Ceux-ci ont ensuite été introduits par Philippe Dagault dans un appareil commercial du type Perkin Elmer instrument Pyris Diamond DSC et ont subi un traitement thermique de 50°C à 400°C, avec des vitesses de chauffage et de refroidissement égales à 10°C/min. Afin de s'assurer de la reproductibilité des résultats, deux batteries différentes pour un même matériau ont été cyclées et analysées. Une cellule de mesure DSC se compose d'un four et d'un capteur intégré avec des positions définies pour les creusets de l'échantillon et de la référence (creuset vide) (voir figure 26). Le profil de DSC (flux de chaleur en fonction de la température) permet de détecter les pics exothermiques associés à la réaction de l'oxygène avec les composants de l'électrolyte, cet oxygène étant libéré par le matériau durant sa dégradation thermique Le profil de DSC indique, d'une part la température jusqu'à laquelle un matériau est stable, et d'autre part l'énergie produite par la réaction entre l'oxygène libéré par la dégradation du matériau et l'électrolyte. 57 Chapitre II – Méthodes et techniques expérimentales utilisées Figure 26 : Représentation schématique d'une DSC II. 4. La diffraction des rayons X sur poudre II. 4. 1. Principe de fonctionnement La diffraction des rayons X sur poudre a été utilisée afin d'identifier la structure cristalline des matériaux synthétisés. Son principe est basé sur l'interaction des rayons X avec les électrons des atomes de l'échantillon. Comme illustré dans la figure 27, lorsqu'un cristal constitué de plans atomiques (hkl) séparés d'une distance dhkl est irradié par un faisceau de rayons X de longueur d'onde λ, celui-ci est diffracté selon un angle hkl à chaque fois que la loi de Bragg est vérifiée : 2dhklsinθhkl = nλ (avec n l'ordre de réflexion et θ l'angle de diffraction). 58 Chapitre II – Méthodes et techniques expérimentales utilisées Figure 27 : Principe de fonctionnement de la diffraction des rayons X Pour permettre un affinement des données de diffraction par la méthode de Rietveld, les diffractogrammes expérimentaux doivent être de très bonne qualité. Pour cela, la durée de l'acquisition, le choix adéquat du pas de la mesure et la préparation de l'échantillon sont importants pour obtenir de bons diffractogrammes. Pour tous les matériaux, la poudre est broyée au mortier et introduite sur un porte échantillon puis arasée à l'aide d'une lame de rasoir. II. 4. 2. Appareillage utilisé Différents diffractomètres de géométrie Bragg-Brentano ont été utilisés, un au CEA et deux à l'ICMCB respectivement : - Un diffractomètre Brüker D8 Advance avec un montage θ-2θ (le porte échantillon est mobile), une anticathode de cuivre et un filtre de nickel pour atténuer le rayonnement Kβ (pas de monochromateur Le diffractogramme est enregistré entre 8 et 80° (2θ) avec un pas de 0,02°, et un temps global d'acquisition de 26 min. - Un diffractomètre PANalytical X'Pert MPD Pro avec un montage θ-θ, une anticathode de cuivre avec la présence d'un monochromateur arrière. L'acquisition des diffractogrammes est faite en 34 min dans le domaine angulaire 8° - 80° par pas de 0,016°. Ce diffractomètre a été utilisé pour réaliser des analyses de routine. 59 Chapitre II – Méthodes et techniques expérimentales utilisées Un to D avec un montage θ-θ (le porte échantillon est fixe), le monochromateur arrière est en graphite avec une radiation Kα (Kα1 et Kα2) du cuivre. Le domaine angulaire est de 10°-120° (2θ), avec un pas de 0,02° et un temps global d'acquisition de 63h. Ce diffractomètre a été utilisé pour réaliser des analyses précises requérant un temps d'acquisition long. II. 4. 3. L'affinement des diagrammes de diffraction La structure des divers matériaux étudiés dans ce manuscrit a été déterminée par l'affinement des diagrammes de diffraction des rayons X. Elle consiste à utiliser la méthode de Rietveld à l'aide du programme Fullprof. Cette méthode consiste à ajuster un profil calculé à celui observé expérimentalement, à partir d'une hypothèse structurale correcte, en tenant compte des paramètres suivants : le fond continu, la position, l'intensité et le profil des raies. Ainsi, le diffractogramme calculé est comparé au diffractogramme observé, et les paramètres du modèle sont ajustés par la méthode des moindres carrés dont l'équation (1) est la suivante : M = ∑ wi (yi − yci )2 (1) Où : - yi représente l'intensité mesurée au pas i du diffractogramme - yci représente l'intensité calculée au pas i du diffractogramme - wi représente la pondération assigné à chaque point du diffractogramme L'affinement est conduit en deux étapes : 1) Full pattern matching (ou méthode de Le Bail) Cette première étape permet d'indexer les raies du diffractogramme et d'affiner les paramètres de maille. En effet, le logiciel Fullprof permet d'effectuer des affinements sans connaitre le modèle structural de la maille considérée, en prenant uniquement en compte le fond continu, la position et le profil des raies. La position des raies dépend de la longueur d'onde du faisceau incident et de la structure du matériau étudié (symétrie, groupe d'espace et paramètre de maille). Les positions des raies vont permettre d'identifier le composé étudié en se basant sur des références de composés connus à l'aide d'une base de données. L'intensité des ies n'est pas reliée à ce stade à un arrangement structural mais est modélisée par une intensité calculée par la relation (2) : yci = ybi + ∑k2 k1 Ik. Ωik 60 (2) Chapitre II – Méthodes et techniques expérimentales utilisées Où : - ybi est l'intensité du fond continu au pas de mesure i - Ωik décrit la forme du pic k à l'angle de Bragg 2θk - Ik est l'intensité intégrée du kième pic Ainsi, les intensités doivent être ajustées en même temps que les autres paramètres décrivant le profil de raies. Le profil des raies correspond à la convolution de la distribution en longueurs d'onde du faisceau incident, de la géométrie du diffractomètre (de la largeur instrumentale) et du profil de diffraction de l'échantillon (taille des domaines cristallins, micro contraintes,). 2) La méthode Rietveld Ces paramètres de maille affinés par la méthode « Full Pattern Matching » vont être ensuite utilisés pour l'affinement global du diagramme de diffraction par la méthode Rietveld. Pour cela, le fond continu, la position et le profil des raies sont affinés de la même façon que précédemment mais l'intensité des raies est cette fois évaluée à partir d'un modèle structural approché. Ainsi, les positions des ions dans la maille sont introduites conformément à l'hypothèse structurale considérée. Les paramètres supplémentaires à affiner sont donc le facteur d'échelle, les positions atomiques, les taux d'occupation et les facteurs de déplacement atomique isotrope des ions dans les différents sites de la structure. Chapitre II – Méthodes et techniques expérimentales utilisées Le diffractogramme est considéré comme la somme des contributions des réflexions de Bragg et du fond continu. L'intensité yci en chaque point i se formule alors : 2 yCi = ybi + Σφ Sφ Σk Jφk. LPφk. Oφf. \|Fφk \|. Ωiφk (5) Où : - ybi est l'intensité du fond continu au pas de mesure i - Sφ est le facteur d'échelle de la phase φ - Jφk est la multiplicité de la ré flexion k - LPφk est le fa cteur de Lorentz - Oφf est le facteur prenant en compte l'orientation p référentielle - Fφk est le fa cteur de structure correspondant à la ré flexion k - Ωiφk est la fonction de profil chargée de répartir l'intensité intégrée de la réflexion k Afin d'évaluer la qualité de l'affinement, plusieurs facteurs dits de qualité (ou de confiance) sont calculés. Ces facteurs s'expriment de la façon suivante : Rp = ∑i\|yi − yci / ∑i yi (6) RBragg = ∑i\|Ii − Ici \| / ∑i Ii (7) R2wp = ∑i wi \|yi − yci \|2 / ∑i wi y2i (8) χ2 = (Rwp /Rexp )2 (9) R2exp = (N − P + C)/Σi wi y2i (10) avec Où : - N est le nombre de points mesurés dans le diagramme - P est le nombre de paramètre affinés - C est le nombre de contraintes Les facteurs de profil pondéré Rwp et de qualité χ2 sont les plus représentatifs des progrès de l'affinement car leur numérateur contient la quantité minimisée par les moindres carrés. Rexp représente la valeur minimale attendue (en tenant compte de la statistique du diagramme expérimental). Rp et Rwp caractérisent l'accord entre les diffractogrammes calculé et observé. 62 Chapitre II – Méthodes et techniques expérimentales utilisées III. Techniques basées sur la quantification des métaux de transition par microscopie et la préparation correspondante des échantillons Cette partie du chapitre traite de la caractérisation de nos matériaux par Microscopie Electronique à Balayage couplée à l'analyse par spectrométries des rayons X par dispersion d'énergie (MEB-EDX) et par microsonde (en anglais Electron Probe Micro-Analysis, EPMA). Cette caractérisation a été indispensable afin de mettre en évidence l'apparition ou non du gradient de concentration et a requis la mise au point d'une préparation d'échantillons adéquate pour évaluer les compositions du coeur à l'extérieur des agrégats sphériques. La technique par polissage ionique (figure 28b) a été retenue car moins agressive qu'un polissage manuel du matériau enrobé dans une résine (figure 28a). En effet, le polissage manuel arrache les particules primaires au contraire du polissage ionique. Comme cela apparait dans les encarts donnés à la figure 28, cet arrachage est observé systématiquement pour chaque agrégat. Les techniques MEB-EDX et EPMA ainsi que la méthode de préparation par cross section polisher vont être décrites dans le paragraphe suivant. Le polissage par cross section polisher a été réalisé en collaboration avec Pascale Garreta au sein de Placamat (Plateforme Aquitaine de Caractérisation des MATériaux). Figure 28 : Images obtenues en mode électrons secondaires d'un oxyde lamellaire du type Li1,2Ni0,2Mn0,6O2 et préparé a) par polissage manuel du matériau enrobé dans une résine, b) par cross section polisher d'une enduction du matériau. La Microscopie Electronique à Balayage en Transmission couplé EDX (STEM-EDX, de l'anglais Scanning Transmission Electron Microscopy) a également été utilisée pour 63 Chapitre II – Méthodes et techniques expérimentales utilisées caractériser les matériaux. Cette technique de caractérisation est exposée dans la suite de cette partie. Elle a demandé une préparation d'échantillon par faisceau d'ions focalisé (FIB, de l'anglais Focus Ion Beam) afin d'obtenir une lame mince en section du matériau. Cette technique récente permet d'obtenir des informations avec des résolutions spatiales de l'ordre du nm, ce qui n'est pas possible avec le MEB-EDX ou l'EPMA dont la résolution est de l'ordre de 1 μm. Cette technique de préparation et d'analyse ne sont pas aisées à mettre en oeuvre, seuls quelques matériaux ont donc été ainsi préparés et analysés. Ces analyses ont ainsi été réalisées au sein de la Plateforme de NanoCaractérisation (PFNC) du CEA Grenoble, au sein du campus d'innovation Minatec, par Alexandre Montani pour la préparation FIB, par Adrien Boulineau pour les analyses STEM-EDX. III. 1. La préparation des échantillons et les microanalyses associées III. 1. 1. Préparation des échantillons par polissage ionique Le cross section polisher Ilion+II model 697 Gatan a été utilisé pour polir les échantillons afin d'en caractériser la coupe par MEB-EDX et EPMA. Cette méthode de préparation des échantillons utilise un faisceau d'ions argon qui vient décaper la tranche de l'échantillon. L'échantillon ou plus exactement un collecteur de courant en aluminium sur lequel est enduit le matériau (sous la forme d'une électrode (matériau actif + carbone + liant) comme décrit dans les paragraphes V. 1, V. 2 et V. 3 du chapitre II) est fixé sur un support en titane au moyen d'une laque à l'argent (voir figure 29). Le décapage par un flux continu d'ions argon est réalisé en 2 étapes. La première étape consiste en un polissage réalisé en 2h à 5 kV et la seconde en un nettoyage à 1 kV. Après polissage, je réalise moi-même des analyses en ligne sur la tranche des agrégats sphériques afin d'étudier de manière plutôt qualitative la nature chimique (par MEB-EDX) et quantitative (par EPMA) des constituants du matériau. Chapitre II – Méthodes et techniques expérimentales utilisées Figure 29 : Images obtenues en mode électrons rétrodiffusés a) de la tranche d'une enduction présentant le matériau et préparée par cross section polisher, et b) l'agrandissement de a) représentant la découpe des sphères dans l'enduction III. 1. 2. La Microscopie Electroniques à Balayage (MEB) associée à l'analyse par Dispersion en Energie des rayons X (MEB-EDX) L'observation de la morphologie des matériaux est réalisée à l'aide d'un microscope électronique à balayage (MEB). L'interaction entre le faisceau d'électrons et la matière donne naissance à divers phénomènes (voir figure 30) dont l'émission d'électrons secondaires, d'électrons rétrodiffusés ainsi que l'émission de rayons X. Ces émissions peuvent donner lieu à la formation d'une image si on dispose du détecteur adéquat. Les images donnant le plus d'informations sur la topographie (taille et relief de la poudre) d'un échantillon massif, avec la meilleure résolution, sont celles par émission d'électrons secondaires. L'imagerie par électrons rétrodiffusés peut être utile pour obtenir des informations sur le contraste de composition (ou de numéro atomique). Le microscope est également doté d'un spectromètre EDX (EDX, de l'anglais, Energy Dispersive X-ray spectroscopy) qui permet d'analyser les rayons X émis par l'échantillon et ainsi de connaitre la composition du matériau étudié. Ainsi, cette technique couplée à l'imagerie MEB permet d'obtenir des cartographies chimiques, des lignes de compositions et des pointés chimiques. Chapitre II – Méthodes et techniques expérimentales utilisées Figure 30 : Emissions de particules lors de l'interaction entre le faisceau d'électrons incident et l'échantillon (poire d'interaction) L'appareillage utilisé est un MEB Jeol 6700F au sein de la plateforme de caractérisation Placamat de Bordeaux. Les conditions d'utilisation sont différentes pour les modes secondaires, rétrodiffusés et EDX : En mode électrons secondaires - Dans ce mode de fonctionnement, plus la tension d'accélération des électrons est faible, plus l'information de surface est privilégiée. Nous avons utilisé une faible tension d'accélération de 5 kV, pour obtenir un meilleur contraste pour l'observation des détails de la surface de l'échantillon. Le courant de sonde est de 3 10-11 A : il n'est pas trop élevé pour améliorer la résolution et éviter la dégradation de l'échantillon mais pas trop faible pour avoir une bonne statistique de mesure. La hauteur de l'échantillon (ou distance de travail) a été choisi relativement faible (8 mm) pour améliorer la résolution. En mode électrons rétrodiffusés - Une tension d'accélération de 10 kV a été choisie. Ces électrons récupérés ont une énergie beaucoup plus importante que celle des électrons secondaires. Ainsi les électrons rétrodiffusés proviennent d'une zone plus profonde de l'échantillon que les électrons secondaires (voir figure 30) donc la tension d'accélération doit être plus importante que celle utilisée en mode électrons secondaires. Le courant de sonde, la hauteur de l'échantillon et l'émission sont identiques dans les deux modes, électrons secondaires et électrons rétrodiffusés. 66 Chapitre II – Méthodes et techniques expérimentales utilisées En mode EDX - La haute tension de 20 kV est choisie pour visualiser toutes les raies caractéristiques utiles des éléments supposés exister dans l'échantillon, en respectant un rapport Ep/Ec compris entre 2 et 3 (Ep est l'énergie primaire et Ec l'énergie de la raie caractéristique la plus énergétique analysée). Dans notre cas, l'énergie la plus élevée correspond à la raie du Ni Kα de 7,5 eV soit un rapport Ep/Ec de 2,67. La statistique de comptage est meilleure avec un fort courant de sonde de 3 10-9 A et une augmentation de la hauteur de l'échantillon à 15 mm permet d'obtenir une plus grande profondeur de champ et donc augmente le signal. III. 1. 3. La microsonde électronique (EPMA) La microsonde électronique ou encore nommé microsonde de Castaing (en anglais Electron Probe Micro-Analysis, EPMA) est une méthode d'analyse élémentaire qui permet de mesurer quantitativement la teneur des éléments contenus dans un matériau. Son principe est le suivant : l'échantillon subit un bombardement électronique et réémet des rayons X caractéristiques de l'élément. Les photons X émis par l'échantillon sous l'impact des électrons sont analysés par des spectromètres à rayons X par dispersion de longueur d'onde (WDS : Wavelength Dispersive Spectroscopy). L'appareil est généralement équipé de plusieurs spectromètres WDS (3 à 5 spectromètres) munis de 2 à 4 monochromateurs, chaque spectromètre correspond à une certaine gamme de longueur d'onde. Cela permet la mesure simultanée des pics de plusieurs éléments. Le détecteur WDS permet donc de séparer les photons X par diffraction sur un cristal monochromateur, et d'en mesurer les intensités à l'aide d'un compteur à flux d'argon. Pour la microsonde, la prépondérance a été donnée au système d'analyse quantitative, alors que pour le MEB, la plus grande importance est donnée au système de formation des images. Le principal avantage de la microsonde est sa grande sensibilité (fort taux de comptage) et une bonne résolution spectrale. Ces qualités de sensibilité et de résolution permettent une bonne détectabilité des pics (bon rapport pic/fond continu). Ainsi la spectrométrie par WDS présente l'avantage d'une résolution spectrale 5 à 10 fois meilleure que celle de l'EDX (utilisé en microscopie à balayage) permettant une meilleure séparation de pics voisins. L'avantage du WDS est également la possibilité de faire des analyses quantitatives sur les éléments très légers à partir du Bore. Tout comme le MEB-EDX, l'appareil permet de faire des profils en ligne de composition quantitatifs, des cartographies X ou des mesures de composition locales (un pointé) sur les matériaux. Chapitre II – Méthodes et techniques expérimentales utilisées L'inconvénient majeur de la microsonde électronique est qu'elle est moins adaptée pour les analyses qualitatives. En effet, le spectromètre WDS nécessite une exploration angulaire relativement longue pour la totalité du spectre, alors que le spectromètre EDX donne instantanément une vue d'ensemble du spectre. C'est pour cela que la microsonde est bien souvent équipée également d'un spectromètre EDX. L'instrument utilisé, situé sur la plateforme PLACAMAT, est la CAMECA SX 100 et les analyses ont été effectuées en collaboration avec Michel Lahaye. Les conditions utilisées pour tous nos matériaux sont une tension de 12 kV et un courant de 20 nA. III. 2. La préparation des échantillons et les nanoanalyses associées III. 2. 1. Préparation des échantillons par faisceau d'ions focalisé (FIB : Focused Ion Beam) La technique FIB va nous permettre de préparer des lames minces afin de pouvoir les utiliser principalement pour le STEM. Préalablement l'échantillon doit être enduit sur une feuille d'aluminium pour une bonne tenue mécanique (comme décrit dans le paragraphe V.2). La première étape consiste à choisir une zone d'intérêt en MEB (dans notre cas, une zone présentant une sphère) et de la protéger en déposant une fine couche de tungstène W (d'épaisseur 1-2 μm) à la surface de l'échantillon successivement par déposition assistée par faisceau d'électrons (limite l'endommagement lié aux ions) et faisceau d'ions (plus rapide et présente de meilleurs rendements). Le dégagement est effectué sur les côtés, devant et derrière la zone d'intérêt pour obtenir une lame « épaisse ». Pour plus de souplesse de manipulation de la partie de l'échantillon à abraser, la lame « épaisse » est extraite (voir figure 31) à l'aide d'un micromanipulateur intégré et attachée à une grille TEM de type Omniprobe par micro collage au tungstène. La suite de la procédure consiste à abraser l'avant et l'arrière de la sphère jusqu'à l'obtention d'une lame mince (typiquement de l'ordre de 100 nm d'épaisseur) afin de pouvoir l'analyser par STEM. Cette étape est la phase d'amincissement dont les conditions varient selon l'épaisseur de la sphère avec un courant variant de 700 à 40 pA et une tension de 30 kV. Pour finir, une phase de nettoyage consiste à réduire la couche amorphe qui se crée de façon inéluctable lors de la phase d'amincissement et qui est liée aux interactions Ion-matière. Cette seconde étape est réalisée successivement à 30, 10, 5, 2 et 1 kV avec des courants de 80 à 10 pA. Ces lames minces ont été réalisées par Alexandre Montani au sein de la PFNC avec l'instrument Dual Beam Carl Zeiss NVision 40. Chapitre II – Méthodes et techniques expérimentales utilisées Figure 31 : Abrasion et extraction de 2 sphères du matériau par la technique FIB III. 2. 2. La Microscopie Electronique en Transmission en mode Balayage couplé en mode EDX (STEM-EDX) Dans la technique STEM (Scanning Transmission Electron Microscopy), le faisceau d'électrons est focalisé sur l'échantillon : on parle alors de sonde. Cette sonde balaye l'échantillon. L'image est formée par détection des rayonnements (électrons ou rayons X) réémis en chaque point de l'échantillon. La résolution en image et en analyse est directement liée au diamètre de la sonde électronique. Les derniers développements permettent aujourd'hui l'obtention de sondes dont la taille est bien inférieure à l'Angstrom. Les meilleures résolutions actuelles sont de l'ordre de 50 pm. Le second intérêt est lié à la possibilité de faire de l'analyse chimique simultanée en analysant soit les rayons X réémis par EDX (STEM-EDX), soit les pertes d'énergie subies par les électrons après avoir interagi avec l'échantillon. Dans un échantillon massif (MEB ou microsonde) le volume analysé est de l'ordre du μm. Ce volume est lié essentiellement à la diffusion électronique (comme visible sur la figure 32a) et ne dépend que peu du diamètre de la sonde. En STEM, l'échantillon étant une lame mince de très faible épaisseur (typiquement 100 nm) il y a peu de diffusion électronique. L'élargissement du faisceau dans l'échantillon est donc très limité (figure 32c). Chapitre II – Méthodes et techniques expérimentales utilisées Figure 32 : Représentation schématique d'un faisceau traversant un échantillon a) massif (la diffusion électronique et l'élargissement faisceau dans la cible rend la résolution spatiale de l'ordre du μm), b) semimince (rendant la résolution spatiale meilleure) et c) dans une lame mince (de l'ordre du nm). C'est pour cela qu'avec un STEM actuel, en routine, les résolutions spatiales associées aux cartographies chimiques sont de l'ordre du nm. IV. Un générateur électrochimique Un générateur électrochimique désigne un dispositif transformant spontanément l'énergie libérée lors d'une réaction d'oxydo-réduction en énergie électrique. On distingue couramment les générateurs électrochimiques rechargeables et non rechargeables : Un générateur électrochimique rechargeable (ou encore secondaire) - C'est un accumulateur qui permet d'être utilisé de manière réversible, c'est-à-dire avec plusieurs cycles de charge et de décharge. Une générateur électrochimique non rechargeable (ou encore primaire) - C'est une pile qui ne peut faire intervenir de système électrochimique réversible. Il ne peut donc pas se charger pour être utilisé une seconde fois. Chapitre II – Méthodes et techniques expérimentales utilisées Les accumulateurs et piles sont composés d'une seule cellule électrochimique. Ces cellules sont ensuite assemblées en série et/ou en parallèle pour constituer des batteries (ou packs de batteries) de façon à obtenir la tension et la capacité de stockage souhaitées. IV. 1. Principe de fonctionnement d'un accumulateur Le principe de fonctionnement d'un accumulateur repose sur l'échange réversible des ions accompagné d'un mouvement des électrons. Pour cela, ce dispositif se compose de deux électrodes (qui se composent elles-mêmes d'un collecteur de courant et d'une entité chimique redox) reliées par un circuit électrique externe via un générateur lors de la charge ou un récepteur en décharge. Les deux électrodes sont le siège de réactions d'oxydo-réduction. Pour que ces réactions puissent avoir lieu, il faut que les ions circulent librement entre les deux électrodes au moyen d'un électrolyte. Afin d'isoler électroniquement le système et d'éviter les problèmes de court-circuit, on utilise une membrane polymère appelé séparateur. Lorsque l'accumulateur fonctionne en charge et en décharge, les réactions électrochimiques mises en jeu sont respectivement les suivantes : En décharge - à l'électrode positive (réduction cathodique) : a Ox1 + n1 e− → b Red1 - (11) à l'électrode négative (oxydation anodique) : c Red2 → d Ox2 + n2 e− (12) En charge - à l'électrode positive (oxydation anodique) : b Red1 → a Ox1 + n1 e− - (13) à l'électrode négative (réduction cathodique) : d Ox2 + n2 e− → c Red2 (14) Dans le cas d'accumulateur au lithium, deux types d'accumulateur existent : les accumulateurs « Lithium-Ion » et « Lithium-Métal ». La figure 33 montre le principe de fonctionnement d'un accumulateur « Lithium-Métal », en décharge pour la figure 33a et en charge pour la figure 33b. Leurs principes diffèrent principalement par les matériaux utilisés au niveau de l'élect négative. En effet, un accumulateur au « Lithium-Métal » se compose d'une électrode négative en lithium-métal alors que dans le cas du « Lithium-Ion », l'électrode négative est constituée d'un matériau d'insertion, tout comme l'électrode positive. Dans la majorité des accumulateurs Li-ion commerciaux, l'électrode négative est du graphite. La technologie « Lithium-Métal » présente un grand inconvénient qui est l'apparition de dendrites [3]. En effet, lors de la recharge de l'accumulateur, le cation vient se redéposer sur l'électrode négative de façon inhomogène. Des filaments dendritiques grandissent au fur et à mesure des cycles de charge et de décharge, pouvant perforer le séparateur et entrer en contact avec l'électrode positive. Ce type de problème entraine des court-circuits au sein de l'accumulateur, et donc à long terme le détériore. Figure 33 : Schéma du principe de fonctionnement d'un accumulateur « Lithium-Métal » a) en décharge et b) en charge V. Tests électrochimiques Les caractérisations électrochimiques des matériaux synthétisés sont réalisées principalement en piles boutons de type CR2032. Les matériaux les plus optimisés ont également été testés en cellule complète de type « sachet souple ». Ce paragraphe présente en plus de la description des différents formats d'accumulateurs, le protocole de préparation de ces accumulateurs. Pour réaliser un accumulateur «Lithium-Métal» ou «Lithium-Ion», le protocole expérimental à suivre est décrit ci-après : - La première étape est la réalisation d'une encre - Cette encre est ensuite déposée sur le collecteur de courant afin de former l'électrode - Après son séchage, l'électrode est découpée et compressée - Pour finir, l'accumulateur est monté en pile bouton ou en « sachet souple ». Chapitre II – Méthodes et techniques expérimentales utilisées V. 1. La réalisation d'une encre L'encre est une solution plus ou moins visqueuse qui contient principalement le matériau actif. Il existe deux types d'encre, les encres préparées par voie aqueuse et celles qui le sont par voie organique. L'encre nécessaire à l'élaboration de l'électrode positive sera réalisée par voie organique afin d'éviter la dégradation du matériau. L'électrode négative en graphite nécessaire aux accumulateurs « Lithium-Ion » est, cette fois ci, pour des raisons de coût et environnementale réalisée par voie aqueuse. De plus, les graphites commerciaux sont maintenant optimisés pour des formulations aqueuses. Par voie organique, on mélange manuellement le matériau actif et le conducteur électronique. Afin d'obtenir une bonne accroche sur le collecteur de ourant, le liant (dilué dans le solvant) est ajouté au mélange. Pour terminer, on ajoute un solvant qui permet d'ajuster la viscosité de l'encre pour une meilleure enduction. Pour les électrodes positives (par voie organique) : - 80% de matériau actif. - 10% de conducteur électronique : Noir de carbone (Super P) - 10% de liant : PVdF (Polyfluorure de Vinylidène) - un solvant organique : NMP (N-méthyl-2-pyrrolidone) Dans le cas d'accumulateur Li-ion, une électrode de graphite est préparée par voie aqueuse. On mélange le matériau actif avec l'épaississant CMC (CarboxyMethylCellulose), le liant SBR (Styrène-Butadiène Rubber) et le solvant. Le SBR permet une bonne flexibilité de l'électrode. On mélange cette préparation pendant 20 min à 4000 tours par minute avec un « dispermat ». Pour terminer, on complète avec le solvant aqueux si besoin. Pour les électrodes négatives (par voie aqueuse) : - 96% de matière active (Graphite Hitachi SMG-HP2) - 2% d'épaississant : CMC (CarboxyMethylCellulose) - 2% de liant: LD417 SBR (Styrène-Butadiène Rubber) - un solvant aqueux : eau distillée Une fois que ces deux mélanges sont bien homogènes, les deux encres sont prêtes à être enduites sur un collecteur de courant en aluminium pour l'électrode positive et en cuivre pour l'électrode négative. V. 2. L'enduction de l'encre et son séchage Quelle que soit la nature de l'électrode, l'encre obtenue est étalée uniformément avec une épaisseur contrôlée sur la feuille en aluminium ou en cuivre servant de collecteur de courant (voir figure 34). C'est la racle qui va permettre de contrôler cette épaisseur. Une fois l'encre enduite sur le collecteur de c , on parle alors d'électrode. L'électrode est enfin séchée dans une étuve à 60°C pendant 24h. Figure 34 : Enduction d'une encre sur un collecteur de courant en aluminium. V. 3. Préparation des électrodes Une fois l'électrode sèche, on la découpe à l'emporte-pièce. Les emporte-pièces peuvent être variables d'un format d'accumulateur à l'autre. Dans le cas des piles boutons, des emporte-pièces standards ronds d'un diamètre de 14 mm sont utilisés. Alors que pour les « sachets souples », ce sont des électrodes carrées de 3535 mm pour la négative et 3232 mm pour la positive. Ensuite, l'électrode est compressée à 10 tonnes afin de la densifier et de favoriser la mobilité des électrons dans l'électrode (soit, une bonne percolation électronique). Les électrodes sont ensuite pesées, puis introduites dans un four à vide de type Buschi®. Une fois séchées, les électrodes sont introduites en boîte à gants pour l'assemblage des accumulateurs en milieu contrôlé. Chapitre II – Méthodes et techniques expérimentales utilisées V. 4. Le montage des accumulateurs V. 4. 1. Les accumulateurs piles boutons Les piles boutons sont assemblées en boîte à gants sous atmosphère inerte d'argon. Le protocole est simple et schématisé dans la figure 35. On introduit dans une coupelle en acier inoxydable avec un joint en polypropylène, l'électrode positive préalablement séchée. Ensuite, on ajoute le séparateur Viledon® servant de réservoir pour l'électrolyte. Et on dépose un deuxième séparateur en Celgard® 2400 d'un diamètre de 16,5 mm. Puis, on imprègne les séparateurs de 150 μL d'électrolyte. Dans le cas de pile bouton de type «Lithium-Métal», on imprègne les séparateurs avec un électrolyte composé de 1M de LiPF6 dissout dans un mélange de carbonate d'éthylène (EC), de carbonate de propylène (PC) et de carbonate de diméthyle (DMC) dans les proportions volumiques 1:1:3 puis on y dépose l'électrode négative en lithium-métal. Alors que pour les cellules de type «Lithium-Ion», on utilise un électrolyte composé de 1M de LiPF6 dissout dans un mélange de carbonate d'éthylène (EC), de carbonate de méthyle éthyle (EMC) et de carbonate de diméthyle (DMC) dans les proportions volumiques de 1/1/1 puis on y dépose l'électrode négative contenant du graphite. L'électrolyte 1M LiPF6 dans EC:PC:DMC (1:1:3) ne peut être utilisé dans les accumulateurs « Lithium-Ion » car le carbonate de propylène (PC) s'intercale dans le graphite par exfoliation. Pour finir, on ajoute une cale en acier inoxydable, un ressort et le capot pour fermer le tout. Figure 35 : Schéma d'assemblage d'une pile bouton Avant d'être testée sur un banc de cyclage, la pile bouton est fermée à l'aide d'une sertisseuse en boîte à gants. A sa sortie de la boîte à gants, elle est nettoyée à l'éthanol afin de retirer toutes traces éventuelles d'électrolyte. Chapitre II – Méthodes et techniques expérimentales utilisées V. 4. 2. Les accumulateurs de type « sachet souple » La réalisation de « sachets souples » (ou encore « Pouch cell » en anglais) a été faite en salle anhydre au CEA Grenoble. Lorsque les électrodes sont sèches, une patte (collecteur de courant) en aluminium (pour la positive) ou en nickel (pour la négative) est soudée aux électrodes. L'assemblage (figure 36b) se fait dans un sachet en aluminium recouvert de polymère (polypropylène). Il y a superposition de l'électrode positive, du séparateur, généralement en polyoléfine PE et/ou PP (de type Celgard® 2400) recouvrant toute l'électrode, et de la négative dans le sachet (figure 36a). Ce dernier est ensuite thermoscellé, laissant uniquement une ouverture pour l'introduction de l'électrolyte dans le « sachet souple ». Le dispositif est séché pendant une nuit. L'ajout de l'électrolyte 1M LiPF6 dans EC/EMC/DMC (1/1/1) en boîte à gants permet l'activation du « sachet souple ». Pour finir, le dernier côté du sachet est ensuite thermoscellé afin de fermer le tout. Figure 36 : a) La photo et b) le schéma d'assemblage d'un « sachet souple » V. 5. Protocole de tests Les accumulateurs (pile bouton ou « sachet souple ») sont ensuite placés sur leurs supports (figure 37) puis branchés sur le banc de cyclage de marque Biologic de modèle VMP (à l'ICMCB) et Arbin BT-2000 (pour le CEA). Toutes les mesures électrochimiques ont été menées en mode galvanostatique pour les piles boutons et en mode potentiostatique pour les « sachets souples ». Ce mode consiste à appliquer un courant constant entre les deux électrodes dans une gamme de potentiel à définir. Ces bornes de potentiel sont généralement de 2,5 à 4,8 V vs Li+/Li correspondant aux bornes de potentiel classique pour un oxyde lamellaire surlithié et le régime de charge/décharge est de C/10 correspondant au courant fourni/imposé à la batterie pour qu'elle soit chargée/déchargée sur une durée de 10 heures. Chapitre II – Méthodes et techniques expérimentales utilisées Ainsi, ce courant est positif lors de la charge et négatif lors de la décharge. La valeur absolue de ce courant (en A) est importante à définir (voir formule 15) puisqu'il correspond à la vitesse à laquelle la pile bouton sera chargée et déchargée. Ainsi, la cellule va être soumise à un programme dans lequel plusieurs variables sont à fixer (les bornes de potentiel, le régime, ) Figure 37 : Les supports pour a) piles boutons et b) pouchs cell (l'étau permettant de presser la pouch cell à 1,5 newton) Le courant (A) à définir lors du test en cyclage est calculé d'après la capacité spécifique Cs (en mAh/g) suivante : Cs = F × x M × 3, 6 Où - F est la constante de Faraday (1F = 500 C ; 1C = 1 A.s) - x est le nombre d'électrons impliqués dans la demi-réaction concernée - M est la masse molaire du matériau d'électrode (g/mol) 77 (15) Chapitre II – Méthodes et techniques expérimentales utilisées Références bibliographiques [1] T. Agilent, « Agilent 720/725 ICP-OES - RELIABLE. PRODUCTIVE. ROBUST. » [En ligne]. Disponible sur: http://www.agilent.com/cs/library/brochures/5990-6497EN%20720-725_ICP-OES_LR.pdf. [2] T. Agilent, « Agilent 700 Series ICP Optical Emission Spectrometers - User's Guide ». [En ligne]. Disponible sur: https://www.agilent.com/cs/library/usermanuals/Public/2301.pdf. [3] K. Nishikawa, T. Mori, T. Nishida, Y. Fukunaka, M. Rosso, et T. Homma, « In Situ Observation of Dendrite Growth of Electrodeposited Li Metal », J. Electrochem. Soc., vol. Chapitre III Synthèses par coprécipitation des différents matériaux du type Li1+xM1-xO2 avec et sans gradient de concentration Chapitre III – Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO avec ou sans gradient de concentration Chapitre III – Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO2 avec ou sans gradient de concentration Chapitre III - Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO2 avec ou sans gradient de concentration. I. Introduction Dans ce chapitre nous nous intéresserons à décrire la méthode par co-précipitation permettant de synthétiser des matériaux présentant un gradient de concentration (GDC) de composition chimique moyenne Li1,1Ni0,27Mn0,47Co0,16O2. Ces GDC sont constitués d'un coeur Li1,2Ni0,2Mn0,6O2 et d'une surface de composition enrichie en Co et en Ni. Au début de ce chapitre nous présenterons le matériau de coeur Li1,2Ni0,2Mn0,6O2, nous verrons qu'il présente de très bonnes performances en cyclage (environ 250 mAh/g sur environ 100 cycles). Néanmoins, comme expliqué dans le chapitre I § II.2, il présente une faible tenue en puissance, il fait également l'objet de réorganisations structurales irréversibles en surface du fait d'une perte d'oxygènes (sa participation aux processus redox le rend instable en surface dans son état oxydé) et de chute continue du potentiel au cours du cyclage (et par conséquent de la densité d'énergie). Le but est donc d'enrichir la surface de Co et de Ni afin de se rapprocher d'une composition LiNi1/3Mn1/3Co1/3O2 en surface, notamment pour pallier aux problèmes de puissance et de perte d'oxygène. Ensuite, nous décrirons le protocole de synthèse que nous avons mis en ainsi que les paramètres clefs (pH, débit, taille du réacteur) qu'il a fallu contrôler pour obtenir le gradient de concentration au sein des agrégats sphériques d'oxydes lamellaires. Puis, nous comparons les résultats obtenus, en termes d'analyses chimiques, de diffraction des rayons X, de microscopie et d'électrochimie, pour les différents matériaux à gradient de concentration ainsi que pour celui de composition homogène (cad sans gradient). Ce matériau permettra d'estimer l'impact du gradient. 81 Chapitre III – Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO2 avec ou sans gradient de concentration II. Synthèse et caractérisation du matériau de coeur du gradient de concentration : Li1,2Ni0,2Mn0,6O2 II. 1. Première étape de la synthèse : la formation du carbonate Ni0,25Mn0,75CO3 La première étape de la synthèse par co-précipitation est la formation d'un carbonate de composition homogène. La synthèse de ce carbonate consiste à utiliser une seule solution initiale à base de sulfates de métaux et de l'introduire dans un réacteur de 500 ml. Afin de réguler le pH, on y ajoute une solution basique à base d'hydroxyde d'ammonium NH4OH et de carbonate de sodium Na2CO3 (qui permet également la formation du carbonate). Cette solution est agitée et chauffée pendant plusieurs heures, avant d'être filtrée pour récupérer le précipité ainsi formé. Celui-ci est lavé dans 3 l d'eau distillée et séché à 80°C pendant une nuit. La solution de sulfates de nickel et de manganèse permet d'obtenir des espèces M2+ en solution et l'ajout du carbonate de sodium permet de précipiter un carbonate de nickel et de manganèse divalents. Les études de ce matériau ont fait déjà l'objet de recherches au CEA et ont montré que le pH optimum est de 7,5 [1]-[2]. Ces résultats sont en bon accord avec la littérature, pH 7-8 pour les matériaux riches en Li et en Mn [1], [3]–[8] alors que le pH est bien supérieur pour les matériaux riches en Ni [9]–[15], comme mentionné dans les tableaux 2, 3, 4, 5 et 6 du chapitre I. La figure 38 montre le diffractogramme obtenu pour le carbonate Ni0,25Mn0,75CO3, indexé dans le groupe d'espace R3̅c. Sur cette même figure sont identifiées les raies (hkl) des pics les plus intenses. Ces données de diffraction montrent des pics larges (à titre d'exemple, la largeur à mi-hauteur FWHM de la raie (104) est de 0,74° (2Cu)) correspondants à un matériau peu cristallisé et/ou dont la taille des cristallites est très petite (130 Å). En effet, en appliquant la formule 16 de Scherrer, ci-dessous, sur la raie (104) du diagramme de diffraction des rayons X, on peut estimer la taille des domaines cristallins soit 130 Å dans le cas des particules du carbonate. D = λ/(βmat. cosθ) Où : - D est l'estimation de la taille du domaine cristallin - λ est la longueur d'onde de la radiation RX - βmat est la largeur intégrale du pic de diffraction 82 (16) Chapitre III – Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO2 avec ou sans gradient de concentration - θ est l'angle de Bragg En toute rigueur, la largeur observée (βobs) d'un pic de diffraction est la somme de la contribution liée au matériau (βmat) et de la contribution d'origine instrumentale (βinstr) : βobs = βinst + βmat Cependant, dans le cas où nous voulons uniquement une estimation de la taille des domaines cristallins, la largeur instrumentale est négligeable donc nous considérons βobs = βmat Figure 38 : Diffractogramme du carbonate Ni0,25Mn0,75CO3 indexé dans le groupe d'espace R3̅c Le tableau 7 compare les compositions chimiques expérimentales, déterminées par des analyses ICP, et théorique du carbonate. La composition chimique expérimentale ainsi proposée, Ni2+0,23Mn2+0,77CO3, est très proche de la composition théorique. L'analyse chimique par ICP a permis de déterminer le rapport molaire Ni/Mn, et de proposer la composition du carbonate en considérant la formule NiII1-yMnIIyCO3. Cette formule a pu être confirmée en réalisant une analyse thermogravimétrique, décrite à la figure 39 et complémentaire de l'analyse ICP. En effet, elle permet de déterminer la quantité de dioxyde de carbone CO2 libérée lors de la décomposition du carbonate, comme décrit ci-après par l'équation 17 : 1 Ni0,23Mn0,77CO3 + 0,1925 O2  0,23 NiO + 0,77 /2 Mn2 O 3 + 1 CO 2 (17) Dans notre cas, le carbonate a subi une montée en température de 2°C/min jusqu'à 900°C, un palier de 7h à 900°C puis une descente rapide à environ 20°C/min. La perte de masse expérimentale est de 36%, elle est cohérente avec la valeur théorique attendue de 37,5%. Nous pouvons donc confirmer la formule du carbonate de Ni0,23Mn0,77CO3. 83 Chapitre III – Synthèses par co-précipitation des différents matériaux du type Li1+xM1-xO2 avec ou sans gradient de concentration Analyse é lément aire par ICP Ni/Mn 0,29 Ni2+0,23Mn2+0,77CO3 0,33 Ni2+0,25Mn2+0,75CO3 Table au 7 : Rapport molaire Ni/Mn obtenu par une analyse élémentaire réalisée par ICP pour le carbonate Ni0,25Mn0,75CO3. Sa composition est estimée en considérant la formule chimique NiII1-yMnIIyCO3, les métaux de transition Ni et Mn étant divalents.	21,200
W1933579528.txt_1	Open-Science-Pile	Open Science	Various open science	2,015	An. gambiae gSG6-P1 evaluation as a proxy for human-vector contact in the Americas: a pilot study	None	English	Spoken	5,965	10,439	Londono-Renteria et al. Parasites & Vectors (2015) 8:533 DOI 10.1186/s13071-015-1160-3 RESEARCH Open Access An. gambiae gSG6-P1 evaluation as a proxy for human-vector contact in the Americas: a pilot study Berlin Londono-Renteria1,9, Papa M. Drame2, Thomas Weitzel3, Reinaldo Rosas4, Crystal Gripping5, Jenny C. Cardenas6, Marcela Alvares7, Dawn M Wesson5, Anne Poinsignon8, Franck Remoue8 and Tonya M. Colpitts1 Abstract Background: During blood meal, the female mosquito injects saliva able to elicit an immune response in the vertebrate. This immune response has been proven to reflect the intensity of exposure to mosquito bites and risk of infection for vector transmitted pathogens such as malaria. The peptide gSG6-P1 of An. gambiae saliva has been demonstrated to be antigenic and highly specific to Anopheles as a genus. However, the applicability of gSG6-P1 to measure exposure to different Anopheles species endemic in the Americas has yet to be evaluated. The purpose of this pilot study was to test whether human participants living in American countries present antibodies able to recognize the gSG6-P1, and whether these antibodies are useful as a proxy for mosquito bite exposure and malaria risk. Methods: We tested human serum samples from Colombia, Chile, and the United States for the presence of IgG antibodies against gSG6-P1 by ELISA. Antibody concentrations were expressed as delta optical density (ΔOD) of each sera tested in duplicates. The difference in the antibody concentrations between groups was tested using the nonparametric Mann Whitney test (independent groups) and the nonparametric Wilcoxon matched-pairs signed rank test (dependent groups). All differences were considered significant with a P < 0.05. Results: We found that the concentration of gSG6-P1 antibodies was significantly correlated with malaria infection status and mosquito bite exposure history. People with clinical malaria presented significantly higher concentrations of IgG anti-gSG6-P1 antibodies than healthy controls. Additionally, a significant raise in antibody concentrations was observed in subjects returning from malaria endemic areas. Conclusion: Our data shows that gSG6-P1 is a suitable candidate for the evaluation of exposure to Anopheles mosquito bites, risk of malaria transmission, and effectiveness of protection measures against mosquito bites in the Americas. Background Globally, an estimated 3.3 billion people are at risk of being infected with malaria [1, 2]. Although the majority of cases and deaths occur in Africa, approximately 170 million people in the Americas live at risk of malaria Correspondence: blondono@uscmed.sc.edu 1 Department of Pathology, Microbiology and Immunology, University of South Carolina, Columbia, SC, USA 9 Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, 6439 Garners Ferry Rd, Bldg 2 Rm C3, Columbia, SC 29209, USA Full list of author information is available at the end of the article infection, with 74 % of cases caused by Plasmodium vivax and 25 % by P. falciparum [3, 4]. Malaria is transmitted to humans when the infective sporozoites, located in salivary glands, are injected into the human skin through the bite of a female Anopheles mosquito. These sporozoites are submerged in mosquito saliva [5] that is used by the mosquito to facilitate blood uptake. Mosquito saliva contains physiologically active components able to counteract blood coagulation, active complement, and the response to hinder the bite injury. Interestingly, some of these salivary components can also elicit and modify immune © 2015 Londono-Renteria et al. 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. Londono-Renteria et al. Parasites & Vectors (2015) 8:533 responses in the vertebrate host and can therefore modify the outcome of vector-transmitted diseases [6, 7]. The gold standard to measure the intensity of malaria transmission is the entomological inoculation rate (EIR), which is defined as the number of infectious Anopheles bites per person in a given time [8]. EIR measurement is highly dependent on the density of human-biting Anopheles [9]. The latter is estimated by trapping methods such as human-landing catches (HLCs) of adult mosquitoes. However, the technique of HLC poses ethical concerns as the human “bait” could be exposed to transmission of malaria and other mosquitoborne diseases. In addition, this trapping technique is only applicable to human adults. It is difficult to extrapolate HLC results to children or to pregnant woman that are the most vulnerable to malaria [10]. As an alternative, catching traps such as the CDC light trap, CDC light trap associated with CO2 or the Mbitrap have been developed [11]. However, studies have shown that these alternative methods have several limitations, like preferentially capturing mosquitoes with higher sporozoite rate and consequently overestimating EIR [12] and can not replace HLC [13]. Therefore, new tools able to evaluate vector–host contact as well as the changes of this contact over time are needed to monitor both population and individual exposure and disease risk. Previous works have shown that the immune response against mosquito salivary proteins, specially IgG antibodies, can reflect the intensity of exposure to mosquito bites as well as the risk of infection for vector-borne pathogens [14–16]. The serological evaluation of this immune response and its association with the exposure to malaria vectors is receiving increasing attention, especially regarding the major malaria vectors in Africa and India, An. gambiae sensu lato and An. stephensi, respectively. Transcriptome analysis of the salivary glands of An. gambiae, identified a protein named An. gambiae salivary gland protein 6 (gSG6), a member of the SG protein family known to be exclusively expressed in the adult Anopheles female [17]. Most of the SG proteins seem to be very well conserved among Old World species, An. gambiae, An. arabiensis, and An. funestus. However, clustal alignment of gSG6 showed similarities with the New World Anopheles species, An. quadrimaculatus and An. freeborni [18, 19]. The peptide gSG6 is restricted to the Anopheles genus based on substantial previous research, which has also demonstrated that specific IgG response to gSG6-P1 could serve as a biomarker for exposure to malaria vectors [2, 17, 19–25]. Moreover, the use of the gSG6-P1 salivary peptide has enabled the collection of relevant data on the efficacy of long lasting insecticide treated nets (LLIN) over time [26] and to compare the effectiveness of individual protection tools such as spray bombs and mosquito coils to LLINs [27]. Page 2 of 8 The development of accurate and sensitive tools to identify variations in vector exposure and malaria risk is important to assess the effectiveness of control efforts. So far, the relevance of antibody response against gSG6P1 has only been evaluated in subjects exposed to bites of Anopheles species in the Old World. The purpose of this study was to evaluate whether gSG6-P1 is a suitable biomarker for exposure to bites of American species of Anopheles, serving as an indicator of the efficacy of individual/collective malaria vector control tools and a proxy for Plasmodium transmission risk in the New World. Methods Ethics statement Universidad de Pamplona, Los Patios Hospital in Norte de Santander-Colombia, approved this study. Additionally, an IRB approval from University of North Carolina at Chapel Hill was granted for the previously described double-blind RCT examining effects of permethrin-treated uniforms on the incidence of tick bites in North Carolina outdoor workers. The Military Hospital and the Chilean Army Health Command in Santiago, Chile, granted the approval for the inclusion of Chilean military personnel. Written consent was obtained from each volunteer. Study participants Chile A sample of Chilean soldiers participating for a period of 6 months in the United Nations Stabilization Mission in Haiti (MINUSTAH) in 2011 were included in the present study (n = 45) (Table 1). They were part of a surveillance project of the Department of Preventive Medicine of the Chilean Military to evaluate the risk of vector-borne disease for peace troops working in Haiti. Samples were obtained from troops that were stationed Table 1 Sample description Country Sample description Sample groups n Chile Chilean soldiers serving in Haiti Before Deployment 45 An. albimanus Return 45 Colombia Healthy subjects Species potential exposure Before traveling 26 After traveling 26 An. albimanus An. darlingi An. punctimacula Febrile patients USA Park Rangers Malaria (+) 7 Malaria (−) 35 Before protective clothing 42 An. punctipennis After protective clothing 42 An. quadrimaculatus Londono-Renteria et al. Parasites & Vectors (2015) 8:533 in Port-au-Prince and responsible for road and bridge building. Daily activities included exposure to swamps, rivers, and flooded roads, as well as diurnal and nocturnal patrolling and convoy security duties in different rural sites. All soldiers received training to avoid mosquito exposure before departure. In Haiti, measures against vector-borne infections included personal use of skin repellent containing 34 % DEET (Ultrathon™, 3M, St. Paul, MN, USA), impregnation of occupational clothes with a permethrin-based repellent, weekly oral dose of 300 mg of chloroquine (base), sleeping in container-type facilities with air-conditioning, and indoor residual spraying and space spraying (adulticiding) with permethrin derivate every 15 days. During off-duty hours troops were authorized to wear sport and casual clothing. Almost all soldiers spent their 15-days UN leave at beach resorts in the Dominican Republic. The main malaria vector in both Haiti and the Dominican Republic is An. albimanus [28, 29]. Serum samples were collected before and after returning from Haiti and kept at −20 °C until processing. Colombia In Colombia, the highest reported altitude for Anopheles species is 2000 m above the sea level (m.a.s.l.) [30]. For this study, we included 26 participants (Colombia population 1), aged 18 to 21 years, living in a malaria free area located at 2300 m.a.s.l. in Pamplona City, but traveling for at least 30 days to endemic areas in the Caribbean Coast of Colombia (States of Guajira, Cordoba, Sucre, Magdalena and Santander), where the main malaria vectors are An. albimanus, An. darlingi, and An. punctimacula [31]. These participants were included in a follow-up study to test reactivity to vector salivary proteins before and after traveling to endemic areas. Participants were bled before and within a week after returning to Pamplona. A second group from Colombia (Colombia population 2) was formed by febrile participants (n = 42) seeking malaria diagnosis and medical services at the Local Hospital of Los Patios and Ocana in the Norte de Santander department, which is a known low malaria endemic area, and at the San Francisco de Asis Hospital in Quibdo in the Choco department, considered the second highest endemic area for malaria in the country. Serum samples were collected and kept at −20 °C until processing. United States We selected 42 North Carolina Parks and Forestry rangers participating in a randomized controlled trial to test the efficacy of long-lasting permethrin-impregnated (LLPI) clothing. Serum samples were collected on time 1 (before implementation of protective clothing) and time 2 (after implementation of protective clothing) from participants Page 3 of 8 of the control group (n = 24) and the test group (n = 18). Aliquots of the samples were kept at −20 °C until testing. Malaria testing by PCR Total DNA was isolated from human samples using the gDNA Blood Kit (Life technologies, Carlsbad, CA, USA) according to manufacturer’s instructions. Plasmodiumspecific PCR was performed using a multiplex PCR for the small sub-unit (SSU) rRNA gene described elsewhere [32]. DNA from in vitro culture of the Haiti I/CDC strain of P. falciparum and a blood sample from a thicksmear P. vivax positive patient were used as positive controls. Antibody detection by ELISA The ELISA conditions followed in this study were published elsewhere [26]. Briefly, 96-well ELISA plates were coated with 100 μL/well of 0.5 μg/ml of gSG6-P1 in coating solution (Kirkegaard & Perry Laboratories, Inc. Gaithersburg, MD) serum samples were incubated at 37 °C for 2 h in a 1/100 dilution. After washing, plates were incubated with 100 μL/well of horseradish peroxidase (HRP)-conjugated goat anti-human IgG (1:1000) antibodies at 37 °C for 1.5 h. Later, each well received 100 μL/well of tetra-methyl-benzidine (TMB) (GeneScript, Piscataway, NJ) and 100 μL/well solution of 1 M phosphoric acid to stop the reaction after 5 min incubation with the TMB. Absorbance was measured at 450 nm. Statistical analysis Antibody concentrations were expressed as delta optical density (ΔOD) calculated for each sample subtracting the mean OD value of the negative control and blank wells from the mean OD value of the duplicates for each sample. Positive control ΔOD values from all plates were recorded and averaged. This average was then divided by individual plate positive control ΔOD values to obtain a “calculation factor” for each plate. To correct for plateto-plate variations, we then multiplied each sample ΔOD value by their respective plate calculation factor to obtain normalized ΔOD’s. The difference in the antibody concentrations between two independent groups was tested using the Mann Whitney U test while the difference between two dependent groups was calculated using Wilcoxon matched-pairs rank test. To determine risk of disease by Odd Ratios (OR), we divided the study sample into two groups with high and low IgG anti-gSG6-P1 antibody levels. As a threshold the median ΔOD of 1.3075 was used. Significant differences were established at p values higher lower 0.05. All statistical testing was performed with Prism version 6.0 (Graph Pad Software Inc.) and STATA™ version 10.1 (Stata Corporation). Londono-Renteria et al. Parasites & Vectors (2015) 8:533 Results and discussion Anti-gSG6-P1 IgG as biomarker of exposure to “New World” Anopheles mosquito bites Estimating mosquito-human contact rate in endemic areas for vector transmitted diseases like malaria, is an important tool to measure the risk of transmission of such diseases. Since saliva form arthropods is injected at the time of transmission, efforts has been directed into discover the salivary proteins able to elicit an immune response that correspond to the level of bites a person has received in a determined period of time. Recently, the protein gSG6 form An. gambiae, has been reported as a useful marker for the degree of exposure to mosquito bites in malaria endemic areas of Africa. Research shows that the gSG6 protein presents little sequence variation within species of the An. gambiae complex (i.e. An. melas, An. quadriannulatu, An. arabiensis). However, sequence comparison of the gSG6 with more distant related species like An. stephensi (Asia) and An. freeborni (America) shows a 67 to 71 % sequence identity [17]. There is not available information on the sequence similarity between gSG6 and the South or Central American Anopheles species An. albimanus, An. darlingi or An. nuneztovari). Nevertheless, salivary transcriptome analysis of the Nyssorhynchus subgenus did not reveal SG6 presence in the salivary content of these species [33, 34]. These reports suggested that a possible reason for this divergence is an independent evolutionary event of the gSG6 protein among anopheline subgenera although they also state that more data was necessary to confirm that hypothesis. In spite of the few available information on the American species homology with gSG6 protein [17, 35], several reports showed the gSG6 protein to be specific to the genus Anopheles. Consequently, we wanted to evaluate whether participants exposed to bites of American endemic Anopheles species develop antibodies specific to gSG6-P1, a salivary antigen validated as a biomarker of exposure to An. gambiae s. l. and An. funestus (main malaria vectors in Africa) bites. To do this, we used the gSG6-P1 peptide to assess exposure to Anopheles bites in healthy participants from Colombia living in a malaria-free area before and after travel to endemic malaria regions (Fig. 1a). In this group, we noticed a significant increase of individual levels of anti-gSG6-P1 IgG after traveling (AT) to malaria areas in comparison to those before traveling (BT) (P = 0.006). A similar increase in specific gSG6-P1 IgG levels was observed in Chilean soldiers after a 6-months stay in malaria endemic areas in Haiti and the Dominican Republic (P < 0.0001; Fig. 1b). These results suggest that IgG response to gSG6-P1 is closely associated with the degree of exposure to the bites of An. albimanus, An. darlingi or An. punctimacula, the malaria vectors described in these endemic areas in Latin America. Page 4 of 8 These results implicate that gSG6-P1 peptide, although based on the Old World An. gambiae s. s. mosquito, can be used to evaluate the level of exposure to bites of New World (Americas) species of Anopheles malaria vectors. Previous studies conducted in Senegal, Angola, and Benin have described a correlation between the level of gSG6-P1 IgG antibodies and the intensity of human exposure to Anopheles bites [21, 22, 26]. Cross-response to gSG6-P1 peptide bases on the fact that gSG6 is a well conserved polypeptide presenting homologues in both the Old and New World Anopheles species [36]. In conclusion, gSG6 polypeptide as well as its gSG6-P1 peptide is suitable as a “bite” biomarker of Old World as well as New World species of Anopheles malaria vectors. IgG specific to gSG6-P1 as a proxy of malaria transmission risk Anti-gSG6-P1 IgG levels were assessed in malaria negative (−) and malaria positive (+) groups of participants residing in malaria endemic areas of Colombia where An. albimanus, An. darlingi, and An. punctimacula are the main vectors. Our results demonstrated that the median level of specific IgG antibodies against gSG6-P1 was significantly higher in the malaria infected group compared to the uninfected group (P = 0.0132) (Fig. 2a). Serum samples in Colombia were collected in two different states: 1) Choco with an annual parasitic incidence (API) of 49 cases/1000 habitants, which in 2010 was classified as a high malaria transmission region [37] and 2) Norte de Santander, which is classified as low transmission area (API: 1.3 cases/1000). Interestingly, the difference in median anti-gSG6-P1 IgG levels between malaria infected and uninfected participants was only significant in participants living in areas of low malaria endemicity (P = 0.0002). In the high transmission area, no significant difference in anti-gSG6-P1 levels was observed (P = 0.1113) (Fig. 2b). We also tested the hypothesis that IgG-gSG6-P1 antibody concentration serves as a proxy for the risk of acquiring malaria. For that, we divided our study subjects into two categories: “low antibody levels” (with a ΔOD value <0.774) and “high antibody levels” (with a ΔOD value ≥0.775). Participants with “high” levels of IgG anti-gSG6-P1 antibodies had a 20.24 times higher risk for acquiring malaria (P = 0.0001) than people with lower levels of those antibodies (Fig. 3) suggesting that more mosquito bites elicit higher anti-gSG6-P1 antibodies and that higher antibody levels are also associated with a higher possibility of being exposed to a bite from an Plasmodium-infected mosquito. This suggests that the level of gSG6-P1 specific IgG could be used as biomarker for the risk of malaria infection. Our data confirm previous studies conducted in different parts of the Old World, in particular a recent study in a low to Londono-Renteria et al. Parasites & Vectors (2015) 8:533 Page 5 of 8 Fig. 1 IgG antibody levels against gSG6-P1 before (BT) and after traveling (AT) to a malaria endemic area. a Healthy civilian form Colombia living in a malaria-free area, traveling to malaria endemic places during vacation. b Military personnel from Chile deployed in Haiti in humanitarian mission. p value denote significance by the Wilcoxon matched-pairs signed rank test moderate malaria transmission area of Senegal, which reported an association between IgG response to gSG6P1 and the status P. falciparum infection of children, even during the dry season [22]. Furthermore, this study showed different levels of anti-gSG6- IgG P1 antibodies in asymptomatic Plasmodium carriers compared to clinical malaria cases. In Kenya, an association between the prevalence of P. falciparum and the levels of anti-gSG6P1 IgG was also described [38]. Another study in Burkina-Faso revealed a positive correlation between malaria incidence and IgG response against the gSG6 recombinant protein [39]. All together, the anti-salivary gSG6-P1 peptide IgG response, previously demonstrated to be a biomarker of exposure to Anopheles bites in numerous contexts and in this study, could also be a useful indicator of malaria exposure and infection. This could also be useful to estimate the malaria risk for nonindigenous populations taking chemoprophylaxis such as the Chilean soldiers of our study, which helps to adapt protection strategies for these populations. In a high transmission situation in Colombia, we found no significant differences in anti-gSG6-P1 levels of malaria (+) Fig. They thus present high levels of anti-gSG6-P1 antibodies, which do not necessarily reflect the risk of malaria transmission. Therefore, for such situations there is a need for more specific biomarkers. It is important to clarify that these findings are applicable for malaria transmission settings in the absence of antimalarial drugs as prophylactic treatments. In such cases, individuals will be still exposed to vector bites with a reduced risk of presenting symptoms and evaluation of the gSG6 antibody levels will be suitable marker for human-vector contact. In the African settings where previous studies with gSG6-P1 took place, entomological records were made to evaluate the abundance of Anopheles species present in these areas. So, it was known that the majority of the Anopheles species present in these areas were potential vectors of malaria. In Colombia, between 70 and 90 % of the mosquitoes captured during entomological surveillance belong to genus An. albimanus, An. darlingi or An. nuneztovari considered the main vectors of malaria in the country [40, 41] while An. albimanus is the most abundant Anopheline specie in the Hispaniola Island [42, 43]. However, If same studies will be undertaken in area where only half of the Anopheles are malaria vectors, we might observe the same non significance between infected and non-infected individuals, as in hyperendemic transmission area, probably because people would receive as many uninfected that infected bites and no association between the level of IgG to gSG6-P1 peptide and the risk of transmission would be observed. Human IgG response to gSG6-P1 for evaluating effectiveness of individual protection against mosquito bites A major application for an immunologically based vector-human contact biomarker is the assessment of the efficacy of vector control methods. For instance, Drame et al. proved that the use of insecticide-treated bednets and spray bombs drastically reduced the IgG levels specific to gSG6-P1 peptide, independently of age or transmission season [22, 27]. Other studies also suggest that the IgG response against gSG6-P1 is a reliable alternative to accurately assess the effectiveness of malaria control methods and surveillance programs in areas of low malaria risk [44, 45]. In this study, we evaluated and compared the IgG-gSG6-P1 levels of US park rangers using LLPI clothing during working hours and control participants wearing untreated clothes before and after the implementation of the strategy. At baseline, both groups presented similar levels of IgG specific to gSG6-P1 peptide. One year after the implementation of LLPI clothing, we observed a significant decrease in the concentration of IgG-gSG6-P1 antibodies in the treatment group (P = 0.0049), whereas no changes were observed in the controls (Fig. 4). Anopheles punctipennis is the most abundant Anopheles species in North Carolina (US), mostly biting outdoors after dusk. However, it will attack humans also during daytime in dense woodlands or close to their daylight resting places, where the park rangers might be performing their work activities. Monitoring the efficacy of individual use of anti-malaria vector tools is very challenging. Previous data suggested that the gSG6-P1 salivary biomarker can Londono-Renteria et al. Parasites & Vectors (2015) 8:533 Page 7 of 8 presence of higher mosquito bite exposure (There has been only one case of malaria in 10 years of mission in Haiti), and that gSG6-P1 is a useful marker to show that in the event of increased exposure to mosquito bites subjects were protected against infection. Conclusion The synthetic peptide gSG6-P1 is useful as a proxy indicator of human-vector contact with American Anopheles species. Further investigation is needed to compare the immune response against the specific mosquito species vectors of malaria in America with the response elicited against gSG6-P1 in areas with different malaria endemicity. It is also a useful tool for the evaluation vector control interventions and devices designed to decrease mosquito bite and disease transmission in human populations. Fig. 4 Level of anti-gSG6-P1 IgG antibodies in park rangers using anti-mosquito bite protective clothing (T) and controls (C) on year 1 (basal antibody levels–before intervention (BT) and follow up levels on year 2 (AT). p value denote significance by the Wilcoxon matched-pairs signed rank test be used to assess and monitor the effectiveness of individual or collective protection tools/strategies against Anopheles mosquito bites [22, 26, 27]. Our study confirmed this by showing that the level of Anopheles bites was significantly reduced in those rangers wearing LLPI clothes. We therefore believe that gSG6-P1 salivary biomarker is a useful marker to assess and monitor the effectiveness of protection measures also against New World Anopheles bites. This raises the importance of personal protection to vector bites in the control of malaria. This is in accordance with previous data showing significant reduction of specific IgG levels, human-Anopheles contact and bites in association with the use of spray bombs [26]. However all personal protection tools do not seem to reduce the level of mosquito bites as pointed by the same authors who described a none-significant effect of the use of mosquito coils or electric fans/air conditioning. In addition, by comparing the efficacy of individual protection tools such as spray bombs and mosquito coils to bed-nets, the same authors indicate that the decreases in Anopheles aggressiveness has essentially been due to the use of bed-nets, meaning that these individual protection tools can be complementary but cannot replace bed-nets in the fight against malaria. In the case of the Chilean troops in Haiti, air conditioning facilities and bednets were used for all the troops at base. Additionally, they use of military uniforms, left neck and wrists exposed so although most of the body is protected these areas can still reached by mosquitoes during the night work. These observations suggest that the use of these uniforms, reinforced by chloroquine treatment, reduced the risk of presenting malaria in the Competing interests None of the authors have a relationship or support that can be perceived as constituting competing interests. Authors’ contributions BLR participated in the study design, acquisition of data and data analysis. PMD, FR and AP helped in the production of recombinant gSG6-P1 as well as the in the study design and data analysis. TW and RR participated in the sample collection of Chilean samples, data analysis and manuscript drafting. CG helped in the acquisition of data. JCC and MA carried out the sample collection in Colombia and helped in the data analysis. DMW and TMC participate in the design of the study and the manuscript preparation. All authors read and approved the final version of the manuscript. Acknowledgements The authors thank the University of Pamplona and Colciencias (Colombia) as well as Dr. Christopher Mores at Louisiana State University (USA) for their support in this study. We also thank Charles Apperson, Meagan Vaughn, Sheana Funkhouser and Steve Meshnick for providing samples. Author details 1 Department of Pathology, Microbiology and Immunology, University of South Carolina, Columbia, SC, USA. 2Laboratory of Parasitic Diseases, National Institutes of Health, NIAID, Bethesda, MD, USA. 3Laboratorio Clínico/Programa Medicina del Viajero, Clínica Alemana, Universidad del Desarrollo, Santiago, Chile. 4 Hospital Militar, Santiago, Chile. 5Department of Tropical Medicine, Tulane University, New Orleans, LA, USA. 6Hospital Los Patios, Los Patios, Norte de Santander, Colombia. 7Hospital Emiro Quintero Canizales, Ocana, Norte de Santander, Colombia. 8Institut de Recherche pour le Développement-IRD, Bouaké, Côte d’Ivoire. 9Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, 6439 Garners Ferry Rd, Bldg 2 Rm C3, Columbia, SC 29209, USA. Received: 19 August 2015 Accepted: 8 October 2015 References 1. Nilsson SK, Childs LM, Buckee C, Marti M. Targeting human transmission biology for malaria elimination. 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https://openalex.org/W4362453482	OpenAlex	Open Science	CC-By	2,023	RDE Calibration—Evaluating Fundamentals of Clustering Approaches to Support the Calibration Process	Sascha Krysmon	English	Spoken	11,599	18,700	Keywords: real driving emissions; cycle-generation; vehicle-specific test scenarios; emission calibration; clustering; data analysis; emission testing; intelligent testing Citation: Krysmon, S.; Claßen, J.; Pischinger, S.; Trendaﬁlov, G.; Düzgün, M.; Dorscheidt, F. RDE Calibration—Evaluating Fundamentals of Clustering Approaches to Support the Calibration Process. Vehicles 2023, 5, 404–423. https://doi.org/10.3390/ vehicles5020023 Sascha Krysmon * , Johannes Claßen , Stefan Pischinger, Georgi Trendaﬁlov, Marc Düzgün and Frank Dorscheidt Chair of Thermodynamics of Mobile Energy Conversion Systems, RWTH Aachen University, 52074 Aachen, Germany * Correspondence: krysmon@tme.rwth-aachen.de; Tel.: +49-241-80-48078 Chair of Thermodynamics of Mobile Energy Conversion Systems, RWTH Aachen University, 52074 Aachen, Germany * Correspondence: krysmon@tme.rwth-aachen.de; Tel.: +49-241-80-48078 Abstract: The topics of climate change and pollutant emission reduction are dominating societal discussions in many areas. In automotive development, with the introduction of real driving emis- sions (RDE) testing and the upcoming EU7 legislation, there are endless boundary conditions and potential scenarios that need to be evaluated. In terms of vehicle calibration, this is leading to a strong focus on alternative approaches such as virtual calibration. Due to the ﬂexibility of virtual test environments and the variety of RDE scenarios, the amount of data collected is rapidly increasing. Supporting the calibration engineers in using the available data and identifying relevant information and test scenarios requires efﬁcient approaches to data analysis. This paper therefore discusses the potential of data clustering to support this process. Using a previously developed approach for event detection in emission calibration, a methodology for the automatic categorization of events is presented. Approaches to clustering algorithms (hierarchical, partitioning, and density-based) are discussed and applied to data of interest. Their suitability for different signals is investigated exemplarily, and the relevant inputs are analyzed for their usability in calibration procedures. It is shown which clustering approaches have the potential to be implemented in the vehicle calibration process to provide added value to data evaluation by calibration engineers. 1. Introduction The constant need to optimize modern passenger cars in terms of sustainability, price, emission reduction [1,2], and quality places high demands on the development processes [3–5]. Constantly updated and intensiﬁed emission standards, such as the Euro 6d introduced in 2017 [6,7] or the upcoming Euro 7 regulations [8], set new framework conditions speciﬁcally for the series calibration process of modern vehicles [9–11]. To ensure legal compliance under all relevant conditions and to increase the efﬁciency in daily use, topics such as base calibration, drivability calibration, emission calibration, or operating strategy calibration are using new approaches in their daily processes in order to achieve zero-impact emission vehicles [2,12–14]. Academic Editor: Nicolò Cavina Received: 20 February 2023 Revised: 20 March 2023 Accepted: 29 March 2023 Published: 30 March 2023 p Innovative approaches aim to reduce costs, shorten the time to market, and reduce the number of prototype vehicles while maintaining or even increasing quality levels. A central topic in this context is the virtualization and simulation of development tasks. Conventional processes are partly replaced and supplemented by simulation-based en- hancements [15]. The use of simulation-based approaches allows the speed of test execution and data acquisition to increase. Initial boundary conditions and changes to setups can be adapted more quickly and soaking phases, for example, can be shortened. However, the resulting potential in data acquisition can only be effectively exploited if the data analysis procedures are adapted at the same time. Copyright: © 2023 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/). https://www.mdpi.com/journal/vehicles Vehicles 2023, 5, 404–423. https://doi.org/10.3390/vehicles5020023 Vehicles 2023, 5 405 In the ﬁeld of vehicle series calibration, this represents a major challenge. In the conventional process, each discipline of series calibration (such as baseline, emissions, drivability, hybrid operating strategy, or OBD calibration) currently performs its own tests, including data acquisition and analysis. The data are usually analyzed manually. For example, each emission test performed is examined by a calibration engineer. Due to lack of time, usually only tests that show exceptional cumulative emissions results are examined in more detail. In the case of an overall good result, no deeper analysis is performed, and speciﬁc potentials that are found in all emission tests but may have only a minor impact on a single test are rarely identiﬁed manually. To support the data analysis, the goal is to develop a methodology for the series calibration process of passenger cars that will assist the application engineer in analyz- ing measurement data, summarizing results, and testing vehicles in speciﬁc use cases. Implementing data analysis processes into the daily procedure of application can: • Increase the overall amount of data that is evaluated, thus making use of all available information; • Increase the overall amount of data that is evaluated, thus making use of all available information; • Increase the speed of measurement evaluations by handling large amounts of data, thus taking full advantage of virtually based test execution; • Standardize the calibration process; p • Increase the quality of vehicle calibration by considering the effects of events that have little impact in a single test but may have a major impact in the daily use of the vehicle over the course of its useful life. • Increase the quality of vehicle calibration by considering the effects of events that have little impact in a single test but may have a major impact in the daily use of the vehicle over the course of its useful life. To develop the individual parts of the proposed methodology, the example of emission calibration is used here. While Section 2 ﬁrst summarizes the overall approach of the new methodology, in Section 3, the focus is on data analysis using clustering approaches. Here, various methods for data clustering are presented and discussed in the context of emission calibration. Hierarchical, partitioning, and density-based clustering approaches have shown promising results in previous evaluations and are therefore discussed in this context. The advantages, disadvantages, and the application of the approaches are presented. Finally, the derived procedure to be implemented in the overall methodology is summarized, and an outlook on its application and the advantages is provided. The paper thus presents the fundamentals of clustering approaches that can be considered for the use in the calibration process. 2.1. State of the Art Within recent years, the virtualization of vehicle tests in the process of vehicle cal- ibration has gained increasing interest [12,16–18]. Throughout the complete calibration, different stages of virtualization from Model-in-the-Loop (MiL) [13] and Software-in-the- Loop (SiL) via Hardware-in-the-Loop (HiL) [19–22] to Engine-in-the-Loop (EiL) [23–25] and Powertrain-in-the-Loop (PiL) [26] are methods to support and/or substitute the con- ventional prototype vehicle and test bench activities; they are summarized as X-in-the-Loop (XiL). The advantage of these virtual test benches is the reduction in prototype costs and their high ﬂexibility for the exchange of components and ambient conditions. Test vehicle soaking phases can be reduced, and the number of tests per day can be increased. In addition, the automation of these processes can further increase the test-per-day frequency and reduce costs in operation. For RDE calibration testing activities, several methodologies have been developed, focusing on ways to increase the system’s robustness and efﬁciency. Majorly, combinations of virtual approaches and dedicated cycle generation are suggested, as on-road tests lack reproducibility and a single cycle, such as the “Worldwide harmonized Light vehicles Test Cycle” (WLTC), or a set of cycles do not offer a high enough variation of test scenarios for real-world usage optimization [27]. A variation of test cycles may be generated with different approaches. Research has been carried out on creating worst-case cycles by maximizing dynamics or combining emission simulations or test bench data with design- Vehicles 2023, 5 406 of-experiments evaluations [28–30]. Thus, synthetical cycles are created that may have high impact on emissions but do not necessarily represent real-world driving behavior. For the representation of such, retracing real routes on a chassis dynamometer or in XiL environments was investigated in [31–33]. Furthermore, mostly Markov-chain-based methods are used to created cycles that represent the statistical driving behavior of a mostly regional database. Generation procedures were described by Kondaru et al. in [34], Balau et al. in [35], and Ashtari et al. in [36]. Förster et al. presented an approach that intended to map a large number of driving scenarios on a statistical data basis [37]. The database was ﬁrst divided into micro-trips. Different driver types were deﬁned by statistical deﬁnition, and test environment conditions were clustered using a K-Means clustering algorithm. With the help of an optimization algorithm, driving cycles were generated that covered a wide test space of real-world conditions. 2.1. State of the Art p While the generation of cycles provides added value to testing activities, Wasser- burger et al. presented a cycle generation methodology in [38] that they later used for an automated calibration process in [27]. The proposed cycles were transformed into operating points of an engine map and used as input values for an optimization algorithm that then adjusted the calibration of speciﬁc functions for all identiﬁed load points. Millo et al. pre- sented a methodology for frontloading base engine calibration using an ofﬂine powertrain model in [39]. In this methodology, fully physical models were used and considered the internal combustion engine as well as the exhaust–aftertreatment system. Using a K-Means clustering algorithm, key points (KPs) for the calibration optimization were identiﬁed by clustering engine operating points of different cycles. A set of features of calibration control variables was deﬁned, and the values were iteratively adjusted to optimize emission behavior and fuel consumption. Furthermore, Meli et al. described a methodology for the automated pre-calibration of ECU functions based on neuronal networks in [40,41]. In this research, a combination of HiL measurements and neuronal network models were used to optimize the base calibration of a lambda control, downstream exhaust temperature model, and knock-control functionality. For the extension of the test space, many different approaches exist in the literature for generating test cycles and testing functions on virtual test benches. Cluster methods are used for cycle generation as well as for targeted automated calibration. However, the new methods for calibration are primarily concerned with automated calibration on virtual test benches. However, this process does not yet meet the standard in series application, yet approaches that describe data analysis for the manual calibration process or even as support for virtual approaches are rarely represented in the literature. The approach presented here is intended to close this gap and create a new basis for data analysis in the conventional calibration process. 2.2. Context of the Proposed Methodology The data-based methodology for vehicle calibration, especially in the context of real driving emission (RDE) calibration, must support the entire calibration process and make optimal use of the collected data. For this purpose, the data of different measurement types, tests, and test environments must be collected and evaluated. The overall methodology into which the clustering approach shall be implemented is initially described here. The general process of the methodology is shown in Figure 1 and is described in detail in [42]. All test environments are used to acquire measurement data. Data analysis by means of event detection and statistical evaluation then refers to the entire database content. The use of the common database enables the continuous use of data from the early phases, focusing on test benches, to the ﬁnal phases in the vehicle. In addition to, e.g., emission tests or simulations, measurements of daily driving operation (robustness measurements), which represent the real usage behavior of the system, are also stored. Vehicles 2023, 5 407 Figure 1. Schematic overview of the RDE application and validation methodology. Figure 1. Schematic overview of the RDE application and validation methodology. Following the data acquisition and pre-processing for the database layout, an event detection is applied to the data containing an emission measurement trace. The event detection automatically identiﬁes the sequences with increased emission intensities, as described in [43,44]. The rule for the deﬁnition of critical intensities and the evaluation of the relevant signals can be adjusted. Thus, the transferability to other topics of interest, such as a hybrid operation strategy, is ensured. Measurements without an emission trace are used for statistical analysis; they can help to prioritize the events with regard to statistical relevance and to evaluate the variety of existing measurement data [42]. Finally, the results of the data analysis are fed back into the testing iterations by means of dataset optimization. They can be tested in dedicated, vehicle-speciﬁc test cycles which are built of the events previously detected and statistically evaluated. p y y To ensure maximum use of the collected data during the development and especially the calibration process, automated analysis is required. In this way, the main advan- tage of virtualized calibration tasks—increasing the speed of measurements and data collection—can be exploited. The goal for automated data analysis is to deliver added value to the calibration engineer. 2.2. Context of the Proposed Methodology The data must be analyzed and visualized in a way that is useful for further evaluation and interpretation by the engineer. Thus, all the beneﬁts of automated analysis should be usable without deep knowledge of data science. The work packages presented in Figure 1 subdivide the process into Identiﬁcation of typical system behavior, Identiﬁcation of signiﬁcant signals & signal proﬁles, Evaluation of statistical safety and Generation of dedicated test cycles. The methodology focused on in this work describes a part of the methods used in the ﬁrst work package, Identiﬁcation of typical system behavior. The subtopics of this work package are shown in Figure 2. Following this requirement, the process of data analysis should be largely automated within the methodology presented in [42]. Vehicles 2023, 5 408 Figure 2. Schematic overview of data analysis steps. Figure 2. Schematic overview of data analysis steps. Using emission calibration as an example, automatic event detection is applied to the measurement data to identify sequences during tests that exhibit increased emission intensity. To further reduce the individual evaluation effort, the events are then processed into clusters. Here, similar events are categorized into groups, which then can be analyzed by the engineer. In this way, not every single event requires evaluation: only reference events of the cluster itself require evaluation. In addition, this allows for the calculation of the overall impact of a weak spot by summarizing the intensity, amount, and statistical relevance. The approach for clustering data will be discussed in its fundamentals in the context of vehicle application within this paper. pp p p In addition to the reduced evaluation effort, with the results of the Identiﬁcation of typical system behavior (Figure 1) an analysis of relevant signals and root causes becomes possible by using clusters. Cluster comparisons can be used in the Identiﬁcation of sig- niﬁcant signals & signal proﬁles (Figure 1) to speciﬁcally identify cluster occurrences of patterns within critical versus not-critical data clusters, as described in [42]. Since the validation process of cycle generation and emission veriﬁcation is highly dependent on the available data diversity as a source for critical behavior, an assessment of the data variation quality is performed in the Evaluation of statistical safety (Figure 1). Finally, the data are used for the generation of vehicle-speciﬁc test cycles, as further described in [45] and in [42] with an updated approach. 3. Results Depending on the type and size of the data as well as the speciﬁc goal to be achieved, different methods for data clustering can be applied. The choice of method depends on the existing knowledge of the available data and the parameters required for the method. In the example of emission calibration, the signals of interest may have many different and complex structures and characteristics. It is difﬁcult to make assumptions in advance about the number, size, or shape of clusters because the weak spots are highly dependent on the vehicle and the dataset currently being used. To add value to the vehicle calibration evaluation process, the clustering approach must be adaptable with little effort from project to project or use case to use case given the wide variation in data. In the context of investigating different clustering approaches, hierarchical, partition- ing, and density-based clustering methods are evaluated. Fourier-transformation-based methods [49–51] previously showed inadequate results due to the often small number of data points within a single event. For usability in the calibration process, clustering results are evaluated in terms of compactness and differentiability, as well as the required prior knowledge of the data. During the investigation, different signals are used as a reference for the calculation of event-to-event distances and thus the clustering itself. This methodology shall be applicable on all kinds of signals to identify signiﬁcant event groups based on any signal. Later, investigations shall be carried out to identify the characteristic features of a dataset or cluster and will be published in a separate paper. Here, each set of clusters is based on the comparison of a single signal. 2.2. Context of the Proposed Methodology p pp As the events show a high diversity of duration and characteristics, a direct comparison of signal traces is not possible. To overcome slight differences in the shapes and durations of the signals, a dynamic-time-warping (DTW) [46,47] approach is used to compare signals of different events. The distance between the reference signal of the event on which the clustering shall be based is then compared with all other events. Thus, a distance matrix is created, showing the distances for any event combination. The clustering approach shall then group all events with similar traces. To evaluate the clustering results, different measures are used. The ﬁrst measure is the Silhouette Score (1). As an intensive measure, it does not require any additional information and is capable of judging the cluster results based only on the categorization. It describes Vehicles 2023, 5 409 the ratio of similarity within a single cluster and the difference towards the others. This is achieved by using the cohesion a for similarity and the separation b for differences: the ratio of similarity within a single cluster and the difference towards the others. This is achieved by using the cohesion a for similarity and the separation b for differences: s(o) = a −b max(a, b) (1) (1) The silhouette s is calculated for each element o. Finally, the Silhouette Score is the average silhouette over all elements o within the cluster. It results in 0 in the case in which there is only one element in the cluster. The cohesion a expresses the average distances of all elements within a cluster; the smaller a is, the more compact the cluster is. The separation b is understood as the average distance of all elements of the neighbor cluster to the element o; high values of b express a high separation. [48] 3.1. Hierarchical Clustering Hierarchical clustering is often used for the categorization of time-based data. Empiri- cal analyses show that the results of this methodology are usually of high quality [52–55]. One of the main advantages is the lack of a need for parameters to create the clusters. Fur- thermore, a complete cluster structure is created by the algorithm. The resulting hierarchy provides initial information about the dataset. For hierarchical cluster methodologies, there are different approaches to create the clusters. Figure 3 shows an overview of different approaches for hierarchical clustering. Figure 3. Overview of approaches for hierarchical clustering. Figure 3. Overview of approaches for hierarchical clustering. Vehicles 2023, 5 410 For hierarchical clustering, a criterion that deﬁnes the distance between clusters is required. In addition to the selection of the algorithm for merging events and sub- clusters, the distance measure also has a major impact on the clustering results. As a main principle, the methodologies categorize events with the closest distance from each other into clusters. Neighboring events are combined into clusters at the ﬁrst step. The clusters are then successively merged as long as the distance between two clusters is below a deﬁned threshold. For the clustering of emission data, four algorithms—Single Linkage, Complete Linkage, Average Linkage, and Ward Linkage—are used. As a source, an event database consisting of 92 critical events for HC emissions (identiﬁed based on the approach described in [44]) is used. The clustering is applied to the engine speed signal. Single Linkage and Complete Linkage use only one element of each cluster to calculate the distance between two clusters. In contrast, Average Linkage and Ward Linkage use all the elements within the two clusters. To create hierarchical clusters, the distance of the events must be known as a stopping rule. In this case, the distance and the number of useful clusters is unknown. For this reason, the Silhouette Score for 2 to 15 clusters is calculated to experimentally investigate an appropriate distance for the emission events. Similar results were observed on further analyses with different signals and events. The results—expressed by the Silhouette Score—are shown in Figure 4. Figure 4. Silhouette Score results for hierarchical clustering approaches. Figure 4. Silhouette Score results for hierarchical clustering approaches. The results for Complete Linkage and Ward Linkage on the right (Figure 4) show similar results, whereas Single Linkage shows very different results—even demonstrating negative values as the number of clusters increases. 3.1. Hierarchical Clustering The negative results on higher amounts of clusters for Single Linkage indicate inconsistencies at the low levels of the created hierarchical structure (the ﬁrst steps of combining events into clusters). The hierarchical creation of clusters does not allow for the later correction of the lower levels. This leads to the phenomenon that mismatches on a low level have a continuous impact on propagating cluster creation, which leads to inhomogeneous results. This effect leads to a reduction in the distances between the clusters. As it only considers single elements from a cluster for the calculation of distances between two clusters, the Single Linkage approach tends to create large clusters with quite Vehicles 2023, 5 411 different signal traces inside. In contrast to the Single Linkage approach, the other approaches show mainly good correlations toward each other and positive results in the Silhouette Score. These results indicate homogeneous clusters with separation to neighboring clusters at each level of the hierarchy. If the number of clusters is too small, the Silhouette Score increases due to the lack of comparison partners. As the Complete Linkage approach tends to form too many small clusters and Ward Linkage tends to form clusters with the same size, Average Linkage is used for further analyses. y For the generation of hierarchical clusters, the use of stopping rules is required. The stopping rules are used to deﬁne the point in the structure at which further clustering should be stopped. As measures to deﬁne stopping rules and to identify the optimal number of clusters for a dataset, the Elbow Rule, Silhouette Score, and Clustering Gain approaches are evaluated. The variance of the cluster content is considered with increasing numbers of clusters, as shown in Figure 5. Using only one cluster results in the highest variance, as all datapoints are included. In opposition to this, the variance is 0 if the number of events equals the number of clusters as each cluster only contains one speciﬁc signal trace. The Elbow Rule deﬁnes the optimum number of clusters by rating the resulting trace with respect to a change in the gradient. In front of the bending point, the variance strongly decreases, meaning that the clusters are becoming more compact. Beyond this point, the variance in the clusters decreases only slightly, meaning that the compactness of the clusters is only slightly optimized [56,57]. 3.1. Hierarchical Clustering Figure 5 shows the results for cluster analyses with two different signals based on 959 emission events. The comparison of the plots of the accelerator pedal position (left) and the voltage of the downstream lambda sensor (right) shows the difﬁculties in clearly identifying the results according to the Elbow Rule. The right proﬁle shows an obvious bending point at ﬁve clusters. The left proﬁle shows a smoother trace and does not allow for the clear identiﬁcation of the point of the optimal number of clusters. Since the bending point is not pronounced enough for some proﬁles, determining the ideal point using the Elbow Rule is not always possible. Figure 5. Deﬁnition of optimum cluster amounts using the Elbow Rule. Figure 5. Deﬁnition of optimum cluster amounts using the Elbow Rule. The Silhouette Score criterion is well-suited for the iterative assessment of the number of clusters. No further information about the data is required, and it is independent of the type of data used for clustering. The application of the Silhouette Score to analyze the cluster quality is shown in Figure 6, using the example of clusters based on the vehicle speed of 92 critical events. While a low number of clusters is desired to best support data analysis, a minimum number of clusters is required to build clusters that adequately represent the characteristics of the signals. With a decreasing number of clusters, the degrees of freedom for separating the data set decrease, resulting in inhomogeneous and large clusters. The local maximum at three clusters is thus interpreted as an outlier. The global maximum is reached at 18 clusters. However, the manual evaluation of the resulting 18 clusters shows a number of clusters that is too high. The local maxima at ﬁve and seven clusters (Figure 7) show the best results when evaluating the clusters manually. Just as with the Elbow Rule, the Silhouette Score does not show a clear optimum in some cases, which further complicates the use of the Silhouette Score as a decision measure in automated approaches. Vehicles 2023, 5 412 Figure 6. Deﬁnition of optimum cluster amounts using the Silhouette Score for vehicle speed. Figure 6. Deﬁnition of optimum cluster amounts using the Silhouette Score for vehicle speed. Figure 7. Hierarchical clustering with 7 (top) and 5 (bottom) target clusters. Figure 7. Hierarchical clustering with 7 (top) and 5 (bottom) target clusters. 3.1. Hierarchical Clustering Clustering Gain∆[58] is a measure that evaluates the increase in similarity within the clusters with a decreasing number of clusters compared to a decreasing inner homogeneity. Using Equation (2), the Clustering Gain uses the average distance between all elements within the dataset d0 and the average distance of all elements d(j) 0 within a cluster j. nj describes the number of elements within the cluster, and j and k represent the total number of clusters. ∆= ∑ k j=1 nj −1 ∥d0 −d(j) 0 ∥2 2 (2) (2) Vehicles 2023, 5 413 Evaluating 10 different signals of the 92 events shows that the Clustering Gain approach underestimates the number of clusters. A reasonable deﬁnition of the optimum number of clusters has shown to work only for a bit signal, which is usually separated in only a small number of clusters. Signal traces with a nearly constant value often represent the average value of several other signal traces that show a higher dynamic within the signal. Such clusters are then combined, leading to the mixing of dynamic and static signal traces. For this reason, signal traces with a low variance and a nearly constant value are separated from the dataset in advance and can then be applied to an individual clustering loop. Figure 8 shows the separation of the static traces of the engine speed. First, the standard deviation is calculated for each signal trace from the 92 events. The percentile that shows a clear jump is determined to deﬁne the maximum value of signal variance for static traces. Here, the 19th percentile is used to separate 18 events with static engine speed traces (Figure 8, right). Figure 8. Separating constant signals from the dataset (right) based on the standard deviation (left). Figure 8. Separating constant signals from the dataset (right) based on the standard deviation (left). In general, the Clustering Gain approach offers a good base for the automatic judging of the cluster hierarchy, although the result is highly dependent on the type and characteristics of the signals used. Filtering is required to use the Clustering Gain approach as a measure of the cluster number. On one hand, hierarchical clustering can be easily applied to arbitrary signals since it does not require signal-speciﬁc parameters. Furthermore, the method is applicable to large datasets, and the computed hierarchy helps to explore the dataset structure. 3.1. Hierarchical Clustering On the other hand, the inﬂexible assignment and errors in low levels of the hierarchy cause the clusters to become inhomogeneous and the distances between individual clusters to become smaller. Another disadvantage is the complex extraction of the ﬁnal clusters from the hierarchy and the determination of the cluster number. Although approaches such as the Silhouette Score and Clustering Gain can determine the optimal number of clusters, this must be done manually and iteratively for most signals. 3.2. Partitioning Cluster Methodologies The K-Means algorithm [59–61] is a widely used partitioning method. The algorithm optimizes an initial partitioning of a dataset based on a cost function. The advantage of K-Means is that elements can change their cluster, and an optimal partitioning of the dataset is therefore possible. The algorithm is scalable on large datasets and always converges, whereas the solution can be a local minimum. The selection of the initial partitioning can be generated manually or randomly. Since manual partitioning requires a large amount of effort, multiple partitions are used in this analysis. Several random initializations of the algorithm are examined, and the best result is considered. Only the determination of one parameter (cluster number) is necessary for the K-Means algorithm. For the cluster centers, synthetic traces are used which average all events in the cluster. The calculation of the cluster center is essential for the optimal assignment of the individual signal traces. The cluster center serves as a representation of the complete Vehicles 2023, 5 414 cluster and must therefore reﬂect as many characteristics of the cluster elements as possible. Therefore, the average trace for the clustering is chosen to be the Barycenter, according to [62]. The Barycenter is a synthetic progression that is calculated using DTW. Figure 9 shows the comparison of two mean trajectories for the same cluster. The cluster consists of 33 similar signal traces for the engine speed from critical emission events. Figure 9. Calculation of a cluster’s mean trajectory. Figure 9. Calculation of a cluster’s mean trajectory. In contrast to the simple averaging of the signals (Figure 9, left), the Barycenter of the signals (Figure 9, right) can compensate the local temporal signal waveforms. Thus, the Barycenter maps the entire cluster better than simple averaging and allows for a more accurate mapping. The ﬂexibility of DTW leads to a difference in length between the simple averaging and the Barycenter. In addition to synchronization, the averaging of signals of different lengths is another advantage of the Barycenter. Thus, the Barycenter reproduces the core information of the cluster and is later able to represent the main characteristic of the feature. As a partitioning method, K-Means requires the speciﬁcation of the number of clusters, which is not known in advance for the signals from emission events. Therefore, two methods for determining the cluster number are investigated. 3.2. Partitioning Cluster Methodologies On one hand, an iterative evaluation of the results is analyzed, and on the other hand, hierarchical clustering is used to determine the cluster number. As a standard criterion for the evaluation of cluster results, the Silhouette Score is used for an iterative determination of the number of clusters. The optimal partitioning is determined by evaluating the cluster results for different cluster numbers and selecting the highest Silhouette Score result. The optimal partitioning is not always achieved at the absolute maximum value of the Silhouette Score. Cluster numbers that do not show the maximum in the trace of the Silhouette Score can provide better results than numbers that show a maximum Silhouette Score. The Silhouette Score for different target cluster numbers is shown in Figure 10, using the engine torque signal as reference. Figure 10. Silhouette Score for partitioning clustering on engine torque with target number variation. Figure 10. Silhouette Score for partitioning clustering on engine torque with target number variation. Figure 11 shows a comparison of the clustering results of 92 events, sorted into ﬁve (top) and two clusters (bottom). The cluster number is usually underestimated, making the clusters inhomogeneous. Vehicles 2023, 5 415 Figure 11. Comparison of cluster results for event torque using 5 clusters (top) versus 2 clusters (bottom). Figure 11. Comparison of cluster results for event torque using 5 clusters (top) versus 2 clusters (bottom). Clustering Gain can automatically determine the number of clusters in the hierarchical clustering. However, the rigid assignment of the data points leads to inhomogeneous clusters. K-Means allows for a ﬂexible assignment and can optimize the clusters. For this purpose, the clusters determined using the Clustering Gain approach are used as initial partitioning for the K-Means algorithm. However, the investigations using a combined approach of K-Means and Clustering Gain generally do not achieve a signiﬁcant improve- ment in the generated clusters. K-Means is not able to perform the partitioning due to the underestimation of the number of clusters by the Clustering Gain algorithm. Due to the possibility of exchanging data points between the clusters and the minimiza- tion of the average distance within the clusters, the K-Means algorithm forms homogeneous clusters when the optimal cluster number is known. However, the determination of the clus- ter number for emission events cannot be automated reliably for all signals. Furthermore, K-Means tries to partition the complete dataset and has no possibility of detecting outliers. 3.3. Density-Based Clustering The hierarchical density-based clustering methodology HDBSCAN combines the advantages of hierarchical and partitioning approaches. First, a hierarchy is created. Then, an optimization algorithm adjusts the clusters by judging the density of the different branches of the hierarchy. This approach enables a hybrid cluster generation process that does not require information about the desired number of clusters. One of the advantages of this technology is its ability to automatically identify outliers in the data. This prevents the mismatch of clusters when trying to sort outliers into the actual patterns. For control of the HDBSCAN methodology, the two key parameters, minimal cluster size Nmin and minimal density Cρ, need to be deﬁned [63]. The minimal cluster size deﬁnes the number of samples (events) that at least need to be assigned to a cluster. Thus, only dense areas with at least Nmin data points are considered Vehicles 2023, 5 416 for the creation of clusters. Small values for Nmin are favored to consider as many points as possible, whereas values that are too small will lead to the overﬁtting of single clusters and subsequently to the creation of many rather small clusters. q y y The minimal density Cρ describes the characteristic of the area that shall be considered for clustering. A minimal number of samples in a speciﬁc area is required to consider this area for the generation of clusters. An area that is not dense enough will not be considered for the generation of clusters. The smaller the number, the more events will be considered for the formation of clusters. A value that is too high will exclude a high number of events and will cause only extremely dense areas to be considered, while a number that is too low will enable the consideration of even outliers and thus might falsify or negatively inﬂuence the compactness of the clusters. p To identify the inﬂuence of both parameters on the clustering of emission events, further analyses are carried out. Test datasets built of critical emission events are created for six different exhaust emission components, whereas different numbers of events are used for each (Table 1). Table 1. Number of events per dataset per emission component. Emission Component HC CH4 CO NOX NH3 PN Number of events 92 172 986 959 1267 2427 Table 1. Number of events per dataset per emission component. Silhouette Score values of > 0.5 represent good results of the HDBSCAN. Here, the number of clusters is not underestimated, and a good ﬁt is shown. For all investigated signals, such a score could be achieved when evaluating the averaged results over all six emission components. The values for Nmin and Cρ used to achieve the results are listed 3.3. Density-Based Clustering Nine emission-relevant signals are selected as base for the clustering approach listed in Table 2. These signals were selected as samples for signals with both a known inﬂuence on the emission intensity and different characteristics. Here, signals with a bit characteristic (bit fuel cut-off) and high (engine torque and downstream lambda sensor voltage) and low (e.g., vehicle speed and temperature of catalytic converter) dynamics and ranges are used. The HDBSCAN parameters Nmin and Cρ are varied to identify the best cluster setup with the least number of outliers. Judgement is carried out by means of the Silhouette Score and visual evaluation. An overview of the selected parameter combinations for the signals is provided for each of the evaluated emission components in Table 3. The generation of clusters is always based on one signal, creating a set of clusters for each of the listed signal and emission component combinations. Table 2. Overview of HDBSCAN parameter evaluation with averaged results. Signal Number Signal Number of Clusters Silhouette Score Outliers 1 Engine speed 14 0.65 36% 2 Vehicle speed 16 0.65 28% 3 Downstream lambda sensor voltage (bank 1) 6 0.61 32% 4 Downstream lambda sensor voltage (bank 2) 5 0.66 36% 5 Bit fuel cut-off 14 1 0.25% 6 Temperature of catalytic converter 4 0.64 8% 7 Engine torque 21 0.6 46% 8 Exhaust gas mass ﬂow 16 0.62 43% 9 Relative air charge 28 0.55 41% Table 2. Overview of HDBSCAN parameter evaluation with averaged results. Silhouette Score values of > 0.5 represent good results of the HDBSCAN. Here, the number of clusters is not underestimated, and a good ﬁt is shown. For all investigated signals, such a score could be achieved when evaluating the averaged results over all six emission components. The values for Nmin and Cρ used to achieve the results are listed Vehicles 2023, 5 417 in Table 3. The quality of the results appears to be independent of the signal and the size of the dataset (Table 1). For signals with low dynamics (e.g., temperature of the catalytic converter), the share of outliers is rather low. Signals with a high share of detected outliers are indicated mainly by those with different levels with respect to their absolute value. For such signals, the algorithm could be re-applied to the events classiﬁed as outliers with an adjusted parameter set to regroup events of different magnitudes. Table 3. 3.3. Density-Based Clustering Overview of results of the HDBSCAN parameter variation. Emission Component Signal 1 2 3 4 5 6 7 8 9 HC Nmin 7 6 6 3 2 7 8 5 3 Cρ 7 6 2 3 2 7 8 5 3 CH4 Nmin 10 7 4 5 2 6 4 12 8 Cρ 10 4 4 5 2 6 4 12 8 CO Nmin 11 7 12 12 2 10 7 11 4 Cρ 11 7 4 2 2 10 7 11 4 NOX Nmin 13 8 13 13 2 15 8 13 5 Cρ 2 2 13 13 2 15 5 13 5 NH3 Nmin 12 9 14 14 2 5 6 15 11 Cρ 2 6 14 14 2 5 6 2 2 PN Nmin 8 30 11 14 3 10 9 11 4 Cρ 8 7 11 14 3 10 2 2 4 Table 3. Overview of results of the HDBSCAN parameter variation. The identiﬁed best-ﬁtting values for Nmin range from 2 to 30, regardless of the size of the dataset. Eight out of nine signals reach the optimum for Nmin ≤15. Exceptions are only observed for signals with low noise in their proﬁle and the presence of ideal clusters. The parameter for the minimal density Cρ shows a good correlation with the minimal cluster size. The absolute differences of Nmin and Cρ are shown in Figure 12 as a histogram and boxplot. For 40 out of 54 sets, Cρ and Nmin result in the same value. For the majority of differences, the value can be rather neglected. Here, the 75th percentile has a value of 2.25, while the 25th percentile and median are both expressed by a difference of 0. High differences can be considered as outliers (judging both the 97.5th percentile or the interquartile range). In addition, further variations of Cρ with a constant Nmin around the optimum value of Cρ are shown to have a neglectable effect. Since the minimum density and minimum cluster size parameters are mostly the same, the parameters are set as equal for further application. This allows for the reduction of the input parameters for HDBSCAN to a single parameter. The methodology used to create the resulting clusters is a further inﬂuencing factor for the HDBSCAN procedure. Clustering can either be performed following the Excess of Mass or the leaf rule. 3.3. Density-Based Clustering As the standard methodology, Excess of Mass shows a tendency to create a smaller number of clusters with a large size. On the other hand, leaf generates small, homogeneous clusters in larger numbers while also allowing for the formation of large clusters if they are present in a homogeneous layout. Since signals for emission events and for vehicle measurements generally have a high variation in their characteristics, the generation of smaller clusters is preferred in this context. Following the leaf method, a larger number of clusters is chosen for use in emission calibration, which can then be concatenated. With the combination of small values for Nmin and the leaf methodology, the process of concatenating micro-clusters directly is supported by a limit of minimal distance between Vehicles 2023, 5 418 clusters. The εClusterSelection can be used to deﬁne the distance between two clusters. Single clusters with a distance smaller than this will be automatically joined. clusters. The εClusterSelection can be used to deﬁne the distance between two clusters. Single clusters with a distance smaller than this will be automatically joined. Figure 12. Evaluation of Nmin and Cρ differences. Figure 12. Evaluation of Nmin and Cρ differences. In summary, HDBSCAN is a reliable method for categorizing emission critical events into clusters of different sizes. The method identiﬁes the typical traces of different signals. As it is independent of the signal traces, the method can be applied to different signal characteristics and can even categorize static traces into separate clusters. By being able to reduce the control parameters to a single factor, its application in the context of a vehicle application offers a promising base. A tendency to form micro-clusters can be observed, but this can be countered by combining multiple micro-clusters using εClusterSelection. 4. Discussion Section 3 demonstrates the applications of three different approaches to clustering data. For hierarchical clustering, the Average Linkage procedure is the best compromise between many small clusters and a few larger ones. An advantage of hierarchical clus- tering is the small number of required settings. Only a threshold value for the distance to combine neighboring clusters with each other as stopping rule is required, which is challenging to deﬁne in the context of vehicle calibration with different signals of interest as there can be a high variety of signals. During the investigation, different approaches are evaluated to provide an automatic procedure to help the engineer to deﬁne this stopping rule. Comparing the Elbow Rule, the Silhouette Score, and the Clustering Gain, all measures show disadvantages in being applied to different signals. As result, hierarchical clustering is an easy application for different signals and is a reliable procedure even for large datasets. However, for daily use in the context of vehicle calibration, the required pre-evaluation of the signals is a major challenge. The evaluation of the data characteristics exceeds the typical analysis procedures of calibration engineers and is therefore not easy to adopt. Using partitioning clustering with the K-Means approach shows a similar disadvan- tage to what is observed for hierarchical clustering. As a control value, the number of clusters to be created must be deﬁned. Due to the wide variety of signal types and traces within the calibration process, a standard target cannot be assigned easily. Similar to hierarchical clustering, the high variance of signals of interest during the calibration process leads to a necessary pre-evaluation of the data. Finding the optimal number of clusters as a control value for the algorithm proves to be signal-dependent and challenging. The HDBSCAN approach is evaluated as a density-based clustering methodology. Here, the variance of the two control parameters, minimal cluster size and minimal density, shows that equal values can be set for both parameters. Vehicles 2023, 5 419 Using the leaf method as a clustering algorithm shows that smaller clusters are created in greater numbers. In contrast, the Excess of Mass methodology creates only fewer but larger clusters. During the analysis and evaluation process, it is observed that a larger number of smaller clusters is preferable. Here, the combination of individual clusters can be easily performed during their visual examination. 4. Discussion In addition, εClusterSelection is used to deﬁne the distance between two clusters as a threshold value below which two clusters are to be combined automatically. An iteration of this threshold can be used after a brief visual examination to reduce the number of clusters automatically if required and if reasonable. The HDBSCAN shows reliable results for different types of signals with different characteristics. The combination of hierarchy-based clustering and the optimization of the results similar to the partitioning approach proves to be the most ﬂexible of the investigated approaches. In contrast to pure hierarchical clustering, the distance used as a stopping rule is not applied globally but only locally. Each combination of clusters within the hierarchy is evaluated regarding the change in density of the clusters and their surrounding area. As long as a combination of clusters leads to a higher separation of clusters from their environment, the combination is performed. The stopping rule is adjusted for each branch. The ability to consider outliers provides the advantage of not falsifying clusters by trying to sort in all the events. Especially in emission testing, outliers can be caused by the behavior of measurement equipment, environmental impacts, or even signal synchro- nization. Comparing the effort of parameter identiﬁcation, HDBSCAN has an acceptable scope in the calibration process. For the most part, the evaluation of reasonable param- eters can be performed in an iterative, automatic manner. Only a small amount of data science knowledge and experience with the approaches is required to evaluate the resulting cluster characteristics. 5. Conclusions New technologies and approaches are being integrated into the vehicle development process to reduce exhaust emissions and address climate change. The use of virtual calibration approaches in RDE optimization and validation can offer major beneﬁts in terms of the safety of different scenarios and speed of testing. However, to make the most of these approaches, the data processing and analysis itself must also be optimized. In order to better utilize the large amount of data required for RDE testing, a method- ology for the automatic pre-analysis of data is introduced. Using the results of previously developed event-detection procedures [43,44], clustering algorithms are utilized to cate- gorize and presort events. This paper discusses the fundamentals of various clustering approaches in the vehicle calibration process. pp p Hierarchical, partitioning, and density-based clustering methods are evaluated. For a sustainable implementation in the calibration process, the appropriate method must be reliable for different signal types, have a high degree of automation, and require little knowledge about the data and control values. Both the hierarchical and partitioning approaches show good results in terms of clustering when the database is well-known. However, with highly varying signal types and unknown populations of events, neither the desired number of clusters nor a maximum spacing of signals to form the clusters is known. Intensive manual data analyses are required to identify the necessary control parameters. As a combination, the density-based clustering approach shows the greatest potential. As a density-based approach, the HDBSCAN algorithm is used. The analysis shows that the two control parameters, minimum cluster size and minimum density, can be set to the same value, reducing the required inputs. Combining this with the leaf method allows for the creation of a higher number of compact clusters of a rather small size. Clustering data with this approach can signiﬁcantly reduce the manual effort required for data analysis. As there is no need to evaluate all events, the automatic categorization of events provides added value for the calibration engineer. The focus can be on the central events of a cluster. In addition, having numerous events with similar signal characteristics Vehicles 2023, 5 420 simpliﬁes a root cause analysis. Further, the impact of a single weak spot can be quanti- ﬁed. The quantiﬁcation of the weak spots themselves provides a tool for benchmarking calibrations of different datasets or vehicles. 5. Conclusions While this paper presents the basic principles for identifying suitable clustering meth- ods, further research will discuss the application of the clustering methods within the calibration process itself. Implementation of density-based clustering in the calibration process will allow for further validation of the methodology with additional data. For fu- ture application, the approach can be transferred to other technologies. Events and clusters regarding consumption optimization or thermal management and derating can be imple- mented for electriﬁed vehicles. For fuel cell vehicles, automated analyses regarding cold start behavior and humidity management as well as aging behavior are feasible. Data will be automatically accounted for so that even minor anomalies are assessed for their overall impact, whereas the current manual approach tends to focus on high-intensity outliers. Author Contributions: Conceptualization, S.K. and J.C.; methodology, S.K. and G.T.; software, G.T.; validation, S.K. and G.T.; formal analysis, S.K. and G.T.; investigation, S.K. and G.T.; resources, S.K. and S.P.; data curation, S.K. and J.C.; writing—original draft preparation, S.K.; writing—review and editing, J.C., G.T., M.D., F.D. and S.P.; visualization, S.K. and G.T.; supervision, S.P.; project administration, S.K.; funding acquisition, S.P. All authors have read and agreed to the published version of the manuscript. Funding: The presented research was carried out at the Center for Mobile Propulsion (CMP) of RWTH Aachen University, funded by the German Science Council “Wissenschaftsrat” (WR) and the German Research Foundation “Deutsche Forschungsgemeinschaft” (DFG). Data Availability Statement: The data presented in this study are available on request from the corresponding author. The data are not publicly available due to the complexity of the analysis which needs guidance for reproduction and conﬁdentiality of root measurements. Conﬂicts of Interest: The authors declare no conﬂict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. 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https://openalex.org/W4211156064	OpenAlex	Open Science	CC-By	2,013	Forecasts covering one month using a cut cell model	J. Steppeler	English	Spoken	6,590	10,633	Forecasts covering one month using a cut-cell model J. Steppeler1, S.-H. Park2, and A. Dobler3 1Climate Service Center, Hamburg, Germany 2National Center for Atmospheric Research, Boulder, Colorado, USA 3Freie Universit¨at Berlin, Institute of Meteorology, Berlin, Germany J. Steppeler1, S.-H. Park2, and A. Dobler3 1Climate Service Center, Hamburg, Germany 2National Center for Atmospheric Research, Boulder, Colorado, USA 3Freie Universit¨at Berlin, Institute of Meteorology, Berlin, Germany Correspondence to: A. Dobler (andreas.dobler@met.fu-berlin.de) Correspondence to: A. Dobler (andreas.dobler@met.fu-berlin.de) Received: 21 December 2012 – Published in Geosci. Model Dev. Discuss.: 25 January 2013 Revised: 22 May 2013 – Accepted: 24 May 2013 – Published: 3 July 2013 Received: 21 December 2012 – Published in Geosci. Model Dev. Discuss.: 25 January 2013 Revised: 22 May 2013 – Accepted: 24 May 2013 – Published: 3 July 2013 Abstract. This paper investigates the impact and potential use of the cut-cell vertical discretisation for forecasts cover- ing ﬁve days and climate simulations. A ﬁrst indication of the usefulness of this new method is obtained by a set of ﬁve-day forecasts, covering January 1989 with six forecasts. The model area was chosen to include much of Asia, the Himalayas and Australia. The cut-cell model LMZ (Lokal Modell with z-coordinates) provides a much more accu- rate representation of mountains on model forecasts than the terrain-following coordinate used for comparison. Therefore we are in particular interested in potential forecast improve- ments in the target area downwind of the Himalayas, over southeastern China, Korea and Japan. The LMZ has previ- ously been tested extensively for one-day forecasts on a Eu- ropean area. Following indications of a reduced temperature error for the short forecasts, this paper investigates the model error for ﬁve days in an area inﬂuenced by strong orogra- phy. The forecasts indicated a strong impact of the cut-cell discretisation on forecast quality. The cut-cell model is avail- able only for an older (2003) version of the model LM (Lokal Modell). It was compared using a control model differing by the use of the terrain-following coordinate only. The cut- cell model improved the precipitation forecasts of this old control model everywhere by a large margin. An improved, more transferable version of the terrain-following model LM has been developed since then under the name CLM (Cli- mate version of the Lokal Modell). The CLM has been used and tested in all climates, while the LM was used for small areas in higher latitudes. Forecasts covering one month using a cut-cell model The precipitation forecasts of the cut-cell model were compared also to the CLM. As the cut- cell model LMZ did not incorporate the developments for CLM since 2003, the precipitation forecast of the CLM was not improved in all aspects. However, for the target area The Cryosphere Open Access downstream of the Himalayas, the cut-cell model consider- ably improved the prediction of the monthly precipitation forecast even in comparison with the modern CLM version. The cut-cell discretisation seems to improve in particular the localisation of precipitation, while the improvements leading from LM to CLM had a positive effect mainly on amplitude. 1 Introduction The cut-cell approach has recently been investigated in a number of two-dimensional test models (see Steppeler et al., 2002; Dobler, 2005; Lock, 2008; Yamazaki and Sato- mura, 2008, 2010; and Walko and Avissar, 2008). Compared to the more common terrain-following coordinate, the cut- cell approach offers a much more accurate discretisation in the presence of orography and avoids a mathematical er- ror occurring with the terrain-following coordinate, when the change of mountain height between neighbouring grid points surpasses the smallest layer thickness. A more de- tailed discussion of this point is given by Yamazaki and Sato- mura (2010) and Steppeler et al. (2006), hereafter referred to as Stal06. Furthermore, the presence of additional metric terms in the terrain-following coordinate transformation adds a truncation error which reduces the model accuracy. Using a three-dimensional cut-cell model and real atmo- spheric data Stal06 were able to show that the cut-cell dis- cretisation had a positive impact on one-day atmospheric forecasts. Using a total of 50 cases with a resolution of 7 km it was shown that the vertical velocity was forecast differ- ently and more realistically by the cut cells as compared to the model using terrain-following coordinates. The precipita- tion forecast was substantially improved and the RMS (root Open Access Open Access Geosci. Model Dev., 6, 875–882, 2013 www.geosci-model-dev.net/6/875/2013/ doi:10.5194/gmd-6-875-2013 © Author(s) 2013. CC Attribution 3.0 License. 2 The cut-cell model The comparison of the cut-cell model with its control ver- sion gives correct information on the potential impact of the cut cells on forecasts. The comparison of LMZ and CLM puts the LMZ at a disadvantage, as LMZ does not beneﬁt from the improvements of the physics scheme since 2003, which are incorporated in CLM. Nonetheless we compare with CLM to make sure that the differences of the forecasts cannot be traced back to the problems of the control model with tropical rain only. The model used is described in detail in Stal06. A few im- provements were introduced with a view towards easy nu- merical experimentation. The time-step is increased to 90 % of the value used in the corresponding terrain-following model version. For comparison this was 25 % in the model runs reported in Stal06. This was achieved by ﬁne-tuning the implicit treatment of the vertical coordinate, the combination of small cells with neighbouring larger ones and the artiﬁcial increase of the volume of small cells. The last model fea- ture was called the thin wall approximation in Stal06 and is described therein, as well as the implicit treatment of the ver- tical coordinate. The technique for cell merging is the same as used by Yamakasi and Satomura (2008), but applied hor- izontally only, as suggested by E. F. Toro (personal commu- nication, 2001). The cell values of the combined cells are obtained by averaging and the time derivatives are then ob- tained using the large cell values. It was checked that these approximations had no signiﬁcant impact on the model fore- cast, when compared to model runs with a smaller time step. In particular the thin wall approximation was only ap- plied when necessary. For example, small grid lengths in the The model improvements leading from LM to CLM con- cern mainly changes in the precipitation scheme to make the model more suitable to different climatic areas. Due to deﬁ- ciencies in the precipitation forecast with the LM, a retuning of the physics scheme and error corrections were carried out, particularly to reduce the too high convective precipitation in the tropics. The changes include the adaption of the autocon- version parameter in the Kessler precipitation scheme and an adjustment of the prediction of the snow phase using a deter- mination of parameters coming from the whole atmosphere rather than from the lower atmosphere only. J. Steppeler et al.: Forecasts covering one month using a cut-cell model J. Steppeler et al.: Forecasts covering one month using a cut-cell model 876 mean squared) of temperature of the one-day forecast was reduced by several tenths of a degree as averaged over 50 one-day forecasts. vertical do not require the cell to be combined with a neigh- bouring one, when treating the vertical coordinate implicitly. The boundary conditions are implemented as described in Steppeler et al. (2002). Because of the short forecast time of one day, Stal06 could only produce small improvements in the temperature and wind ﬁelds. The question arises, if for longer forecasts the cut-cell discretisation has a stronger impact on forecasts. Eventually, this question will have to be answered using a large number of consecutive forecasts and an up to date model physics scheme. From the standpoint of the user the LMZ is identical to the LM model (Steppeler et al., 2003) and its climate simulation version CLM (Rockel and Geyer, 2008). In particular the in- put and output ﬁles of LMZ are obtained by interpolating the z levels to the same terrain-following levels as used in the terrain-following models LM and CLM. The LM will be referred to as the control model and differs only by the vertical coordinate from the cut-cell model LMZ. In particular the physical parameterisations are the same for both models and are described in Steppeler et al. (2003). These include a multilayer soil model (see Schrodin and Heise, 2002), a radiation scheme following Ritter and Geleyn (1992), and a Kessler-type microphysic scheme (Kessler, 1969). The Tiedtke (1989) parameterisation scheme is used for convection and a diagnostic TKE (tur- bulent kinetic energy) scheme for the boundary layer. The physics and interpolation options are as in Stal06. No tuning of the physics scheme was done, but rather the physics are taken over unchanged from the control model. The intention of the present paper is to show the impact of the cut-cell discretisation for longer integrations. In Steppeler et al. (2011) a similar study was carried out, but for a smaller area and ﬁve days only. The larger area used herein makes the solution less dependent on lateral boundaries. Additionally, a comparison to an up-to-date version of the terrain-following model is included. A set of ﬁve-day forecasts is produced covering January 1989 with six forecasts. This is a further step towards a test of cut cells for long-time forecasts and cli- mate runs. J. Steppeler et al.: Forecasts covering one month using a cut-cell model For the ﬁnal test of the method, the authors aim to run 100 ﬁve-day forecasts using mesoscale resolution (about 7 km) and an area of a similar size as in the present paper. Lateral boundary values from ERA-interim data are used. Such model set-up is often used for model performance eval- uation in climate impact studies. This will also give a ﬁrst indication of the usefulness of cut cells for such studies. The model area has been chosen to see a strong orographic im- pact. It includes the Himalayas and a large area downwind, including southeastern China, Korea and Japan. In this target area we expect a strong impact of the cut-cell discretisation. The control model and its cut-cell version were developed from an older version of the LM, as described in more de- tail in Stal06. The results, in particular concerning precipi- tation, have improved since then (Rockel and Geyer, 2008). The model including these improvements is called CLM. The LMZ and its control model LM in comparison do not include these improvements and involve no tuning of the physics scheme at all. The older LM used for control purposes was in its time not used in the tropics and did not involve changes of the physics scheme coming from its hydrostatic predecessor. Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. www.geosci-model-dev.net/6/875/2013/ J. Steppeler et al.: Forecasts covering one month using a cut-cell model 6 877 ppeler made to enable the model to run in climate mode (i.e., with longer integration time), the use of the NetCDF ﬁle format for output in order to facilitate the transfer of the model to different computer platforms, the changing of output name conventions to support a large number of output ﬁles and a restart option. made to enable the model to run in climate mode (i.e., with longer integration time), the use of the NetCDF ﬁle format for output in order to facilitate the transfer of the model to different computer platforms, the changing of output name conventions to support a large number of output ﬁles and a restart option. Fig. 1. (a) The 10 m wind in m s−1 as forecasted for 5 days from 21 January 1989 by the terrain-following model LM; (b) as (a), for the 0 day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. Fig. 1. (a) The 10 m wind in m s−1 as forecasted for ﬁve days from 21 January 1989 by the terrain-following model LM; (b) as (a), for the zero-day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. The model runs reported in this paper are ﬁve-day runs with starting dates 01, 06, 11, 16, 21, 26 January 1989. The model area can be seen from Fig. 1. The horizontal resolution is 0.25◦, roughly 25 km. There are 521 points in east–west direction and 321 in north–south. The model set-up uses 31 layers. The layers 20 and 25 (counted from the top) are used for veriﬁcation. These levels correspond to about 800 and 850 mb for points over the ocean. For the one-day forecasts reported in Stal06 the orogra- phy was ﬁltered for the control model and unﬁltered for the cut-cell model LMZ, as the control model would not pro- duce reasonable results otherwise. The LMZ can run without orographic ﬁltering. The results reported in this paper, how- ever, were obtained with orographic ﬁltering for both model versions, in order to have an exact control model. Therefore a potential beneﬁt of using the more realistic unﬁltered orog- raphy with LMZ is not investigated here. For the model do- main used in this paper, the maximum change of mountain height between neighbouring grid points is 2885 and 1654 m for the unﬁltered and the ﬁltered orography, respectively. J. Steppeler et al.: Forecasts covering one month using a cut-cell model 6 Us- ing a horizontal resolution of 0.25◦this results in relatively small maximum terrain slopes of less than 7◦and less than 4◦for the unﬁltered and the ﬁltered orography, respectively. Fig. 1. (a) The 10 m wind in m s−1 as forecasted for 5 days from 21 January 1989 by the terrain-following model LM; (b) as (a) for the Fig. 1. (a) The 10 m wind in m s−1 as forecasted for ﬁve days from 21 January 1989 by the terrain-following model LM; (b) as (a), for 3 Results When discussing the results we will refer to the cut-cell model as LMZ and to the terrain-following control model as LM. Where a comparison on model levels is carried out, the terrain-following levels are used, as the output of LMZ is interpolated to these levels. Figure 1 shows ﬁve-day forecasts from 21 January of the wind-component u at 10 m height for LMZ and LM. As an indication of the veriﬁcation the zero-day forecast from 26 January is given (for LM). Both forecasts and the veriﬁca- tion show the northern and southern trade wind systems with easterly surface winds. Weak winds prevail north and south of the two trade wind systems and in the convergence zone between them. Strong westerly winds are seen in the north- eastern corner of the forecast area, being associated with cy- clonic activity. For the case shown in Fig. 1 the LM forecast has stronger westerly winds in the tropical convergence zone than the LMZ forecast and the veriﬁcation. Fig. 1. (a) The 10 m wind in m s−1 as forecasted for 5 days from 21 January 1989 by the terrain-following model LM; (b) as (a), for the 0 day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. Fig. 1. (a) The 10 m wind in m s−1 as forecasted for ﬁve days from 21 January 1989 by the terrain-following model LM; (b) as (a), for the zero-day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. Yamazaki, H. and Satomura, H.: Nonhydrostatic atmospheric mod- eling using a combined cartesian grid, Mon. Weather Rev., 138, continuous cyclonic activity, with a corresponding variability of the westerly wind. 3932–3945, 2010. The impact of the cut-cell discretisation is strong. Some differences between the LMZ and LM forecasts are typical. These are a stronger westerly wind patches embedded in the convergence zone for LM and for the LMZ stronger wester- lies in the northeast of the model area and less noisy ﬁelds. These wind systems are highly variable in time (plots not shown). The trade wind zones can be a narrow band or rather wide, as shown in the example in Fig. 1. For the month of January 1989 the northeastern corner in the model area shows 2 The cut-cell model Details on the changes to the precipitation scheme are given in Seifert and Crewell (2008). In addition to this, technical changes were Geosci. Model Dev., 6, 875–882, 2013 www.geosci-model-dev.net/6/875/2013/ J. Steppeler et al.: Forecasts covering one month using a cut-cell model 6 www.geosci-model-dev.net/6/875/2013/ J. Steppeler et al.: Forecasts covering one month using a cut-cell model The use of model initial ﬁelds for veriﬁcation is problem- atic as the data assimilation derives ﬁelds also in areas with no observations. Therefore it is difﬁcult to assess the accu- racy of the data used for veriﬁcation. In particular the vertical velocity ﬁeld w is obtained as a model ﬁeld at t = 0 (Step- peler et al., 2003), with no direct measurements of w being used. Therefore it is desirable to compare directly with ob- servations. Most readily available are precipitation data. As precipitation depends strongly on the vertical velocity, pre- cipitation veriﬁcation can be seen as an indirect veriﬁcation of w. Here, we use the monthly, global gridded precipitation dataset from GPCC (Global Precipitation Climatology Cen- tre; V5, Schneider et al., 2013) with a grid resolution of 0.5◦ over land and from GPCP (Global Precipitation Climatology Project, V2.1, Huffman et al., 2009) with a grid resolution of 2.5◦over the oceans as reference. To merge the data to a common grid, the GPCP precipitation over the oceans has been bi-linearly interpolated to the GPCC grid. The differences of the forecasts are rather large. The biggest improvement in the temperature forecasts can be seen in the cold area north of 40◦N. Here, some artiﬁcial east– west stripes of cold air appear with LM which are not visi- ble with LMZ or in the analysis. Generally, the shape of the cold area with LMZ is in better agreement with the analy- sis than that from LM. However, the amplitude is often too strong with LMZ, but too weak with LM, for instance for the cold semi-circular area in the southwest. As can be seen by the temperature bias (Table 1) for a northern region close to Mongolia from 105 to 115◦E (Fig. 2), the results with LMZ there are colder but closer to the analysis than with LM. The performance over the rest of the area differs and the forecasts there are of similar quality. Figure 3 shows the ﬁve-day forecasts of the vertical ve- locities with starting date 21 January 1989. Again the zero- day forecast from 26 January (for LM) is used for veriﬁ- cation. For the preparation of this initial ﬁeld, see the LM model documentation (Steppeler et al., 2003 and references therein). J. Steppeler et al.: Forecasts covering one month using a cut-cell model 878 Table 1. Temperature and precipitation biases of LMZ and (C)LM for selected areas. Some investigations were done concerning the noisy w ﬁeld with LM and the associated heavy rain. These are sum- marised here without showing all corresponding diagrams. At the initial time LMZ and LM have very similar w ﬁelds, which for the LMZ forecast are evolving continuously and verify reasonably with the w from the analysed data. For LM large differences appear after the digital ﬁlter initialisation and are also seen in adiabatic runs. The digital ﬁlter initialisa- tion creates large-scale differences between LM and LMZ in the vertical velocity ﬁeld, which are not localised near moun- tains. They occur for example in a large area downwind of Madagascar. Table 1. Temperature and precipitation biases of LMZ and (C)LM for selected areas. Area Bias (LMZ) Bias (LM) T −0.3 K 4.2 K Bias (LMZ) Bias (CLM) 1 −20 mm 30 mm 2 −15 mm 75 mm 3 −15 mm −45 mm 4 −5 mm 15 mm For diabatic runs the strong vertical velocities with LM create heavy precipitation, particularly in the tropical belt. This again creates even higher vertical velocities. Apparently the creation of the noisy structures over the whole model area after ﬁve days is caused by ampliﬁcation of rising motion using the energy source of the warm tropical ocean. In the course of the ﬁve-day forecasts the high amplitude features of the w-ﬁeld spread to the whole model area and cause in- creased rainfall rates everywhere. The increased noise level of the forecasts is of a scale of 100–500 km. With a resolution of 25 km this consists of well resolved structures. The difference of LMZ and LM forecasts concerns all model levels. We are in particular interested in the area of cyclonic activity downstream of the Himalayas. Our target area is 30 to 40◦N and 100 to 140◦E. Figure 2 shows the ﬁve-day forecasts for all six cases. The temperature for out- put level 20 is given, corresponding roughly to the 800 mb surface over the ocean. The forecasts of LMZ, LM and the zero-day forecast for the target date for LMZ (veriﬁcation) are shown. Note that the model reaction is very different for precipi- tation and temperature. While the differences in precipitation are already large after the ﬁrst day, the differences for tem- perature take about ﬁve days to build up (not shown). www.geosci-model-dev.net/6/875/2013/ www.geosci-model-dev.net/6/875/2013/ Geosci. Model Dev., 6, 875–882, 2013 www.geosci-model-dev.net/6/875/2013/ J. Steppeler et al.: Forecasts covering one month using a cut-cell model (a) Temperature for the 5 day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the 0 day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T) used to calculate the temperature biases. Fig. 2. (a) Temperature for the ﬁve-day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the zero-day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T ) used to calculate the temperature biases. Fig. 2. (a) Temperature for the 5 day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the 0 day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T) used to calculate the temperature biases. Fig. 2. (a) Temperature for the ﬁve-day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the zero-day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T ) used to calculate the temperature biases. precipitation south of the Himalayas and precipitation over Sri Lanka and Thailand, which are dry with CLM. at some places better for CLM than for LMZ. However, de- spite the lack of these improvements in the LMZ, the cut- cell model performs better than the CLM at some places, as can be seen from the precipitation biases given in Table 1. Forecast differences in favour of CLM are a tendency of the LMZ to predict light rain in areas which are dry. While CLM correctly produces extended areas of no precipitation, LMZ often produces light rain for such locations. This concerns parts of the Arabian Peninsula, India and Australia. This is a known error of older models and the expression “Socialist precipitation” has been coined for this. J. Steppeler et al.: Forecasts covering one month using a cut-cell model Both forecasts and the veriﬁcation show a band of rising motion in the tropical convergence zone, which in the east of the area is split into three branches. One of them is reaching far north into the vicinity of Japan. For the LM run the large-scale features are obscured by small-scale noise of rather high amplitude, which is present everywhere and is strongest over the high mountains and in the tropics, particu- larly downwind of Madagascar. These strong vertical veloc- ities are responsible for heavy rain with LM. The forecasted vertical velocities for LM verify much worse than those of LMZ as compared to the initial ﬁelds. Figure 4 shows the accumulated precipitation for the LM, the LMZ and the CLM forecasts and the observations for the whole month of January 1989. The results from LMZ are almost everywhere more accurate than those from LM, with the latter being generally double the observed values. This again highlights the error in the old LM version resulting in much too high convective precipitation in the tropics. Note that the retuning of the physics and error corrections mentioned in Sect. 2 are not yet available in the LMZ. The LMZ differs from the LM by its vertical coordinate only. The CLM clearly beneﬁts from the improvements compared to LM. This explains that the amplitudes of precipitation are Geosci. Model Dev., 6, 875–882, 2013 www.geosci-model-dev.net/6/875/2013/ J. Steppeler et al.: Forecasts covering one month using a cut-cell model 879 Fig. 2. (a) Temperature for the 5 day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the 0 day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T) used to calculate the temperature biases. Fig. 2. (a) Temperature for the ﬁve-day forecast from 1 January 1989 of level 20 for the terrain-following model LM (left), the cut-cell model LMZ (right) and the zero-day forecast from 6 January 1989 (“veriﬁcation”, middle); (b–f) as (a), for dates 6, 11, 16, 21, 26 January. The black box in the top left corner denotes the area (T ) used to calculate the temperature biases. Fig. 2. J. Steppeler et al.: Forecasts covering one month using a cut-cell model The anomaly correlation is calculated as Fig 3 (a) Vertical velocity w for level 20 of the ﬁve day forecast Corr(model) = 6P t=1 simt −sim reft −ref s 6P t=1 simt −sim 2 6P t=1 reft −ref 2 . Corr(model) = 6P t=1 simt −sim reft −ref s 6P t=1 simt −sim 2 6P t=1 reft −ref 2 . To summarise, the positive impact for bias and anomaly correlation is in favour of the LMZ for all selected locations as can be seen in Tables 1 and 2. This improvement is present even in comparison with the state-of-the-art model CLM. Note that no quantitative comparison with the LM model was done for precipitation, as the LM precipitation is too high al- most everywhere. Fig. 3. (a) Vertical velocity w for level 20 of the ﬁve-day forecast from 21 January 1989 for the terrain-following model LM; (b) as (a), for the 0 day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. Fig. 3. (a) Vertical velocity w for level 20 of the ﬁve-day forecast from 21 January 1989 for the terrain-following model LM; (b) as (a), for the zero-day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. J. Steppeler et al.: Forecasts covering one month using a cut-cell model This clearly indicates that for operational applications the physics of LMZ needs to be modernised. The largest differences are in the target area downwind of the Himalayas: southeastern China, Korea and Japan. The LMZ gives a better distribution of precipitation as compared to CLM. The precipitation is correctly concentrated in the south of China. Korea and Taiwan get precipitation and the rain over Japan is concentrated in the west of the country. These are differences involving a large area and they indicate that the mathematically more correct treatment of mountains with the LMZ has a considerable impact for prediction and climate simulation purposes. It is interesting that over dry ar- eas in Arabia and India the vertical velocity is negative for LMZ and LM. This again may be seen as an indication that the LMZ model needs an improvement of the physics scheme such as the changes leading from LM to CLM. The rain produced south of the Caspian Sea is better with CLM as well. However, when a feature over land is pre- dicted both by LMZ and CLM, its position and shape is of- ten better with LMZ as for instance the banded structure of We leave it to the reader to ponder the forecast differ- ences for the medium sized tropical islands. Over the oceans, www.geosci-model-dev.net/6/875/2013/ www.geosci-model-dev.net/6/875/2013/ Geosci. Model Dev., 6, 875–882, 2013 J. Steppeler et al.: Forecasts covering one month using a cut-cell model 880 The dry areas of the ocean are too large with CLM and the extension of precipitation into the Arabian Sea is better with LMZ. Further, the tropical precipitation belt is smoother with LMZ, which appears to be correct. However, observations over the oceans are known to include biases (Huffman et al., 2009) and are available at low resolution only (2.5◦in this case), resulting in smooth ﬁelds when interpolated to a ﬁner resolution. Fig. 3. (a) Vertical velocity w for level 20 of the ﬁve-day forecast from 21 January 1989 for the terrain-following model LM; (b) as (a), for the 0 day forecast from 26 January (“veriﬁcation”); (c) as (a) for the cut-cell model LMZ Fig. 3. (a) Vertical velocity w for level 20 of the ﬁve-day forecast from 21 January 1989 for the terrain-following model LM; (b) as (a), for the zero-day forecast from 26 January (“veriﬁcation”); (c) as (a), for the cut-cell model LMZ. The authors want to emphasise that the present study is still an impact study. For a proper evaluation of the cut-cells method, a larger sample of simulations and mesoscale res- olution is necessary. Currently, the authors aim to run 100 ﬁve-day forecasts using about 7 km resolution. The analysis of these simulations will include more quantitative veriﬁca- tion. However, some simple scores have been computed to give quantitative meaning to the model differences. The areas for the computation were selected as places where the posi- tion of features was different in the forecasts and are shown in Figs. 2 and 4. For precipitation we extracted the bias for four areas lying in the shoulder of the Himalayas and east- ern China, and for temperature an area close to Mongolia has been chosen. The locations for the computation of the tem- perature anomaly correlations were selected in the lee area of the mountains, where the impact of cut cells is expected to be strongest. The bias is calculated as Bias(model) = sim −ref, where sim are the model values and ref the reference values. For precipitation the mean is taken over the areas shown in Fig. 4. For temperature the mean is taken over the area shown in Fig. 2 for each of the six ﬁve-day forecasts and over the single time values. 4 Conclusions The cut-cell discretisation removes major numerical errors near mountains. It was shown that the impact of this scheme for ﬁve-day forecasts is considerable. The analysed verti- cal velocities verify reasonably well with the cut-cell LMZ model and not very well with the terrain-following LM model. After the digital ﬁlter initialisation the vertical veloc- ities of the two model versions differ on a global scale, with the amplitudes for the tropical precipitation are still some- what too high for both LMZ and CLM, with the maximum reaching often 600 mm, where 400 to 500 mm are observed. Mostly, the overestimation is higher in LMZ than in CLM. However, east of the Philippines LMZ correctly predicts strong precipitation on the sea and immediately at the coast, while CLM has the precipitation a bit away from the coast. www.geosci-model-dev.net/6/875/2013/ www.geosci-model-dev.net/6/875/2013/ Geosci. Model Dev., 6, 875–882, 2013 J. Steppeler et al.: Forecasts covering one month using a cut-cell model 881 The forecasted accumulated precipitation for the whole January 1989 using the terrain-following model LM; (b) as (a to the GPCC (land) and GPCP (ocean) dataset; (c) as (a), for the cut-cell model LMZ; (d) as (a), for the terrain-follow s current version. For (c) and (d), the same colour scheme as for (b) was used. The lines and the numbers 1 to 4 in (b) d to calculate the precipitation biases Fig. 4. (a) The forecasted accumulated precipitation for the whole January 1989 using the terrain-following model LM; (b) as (a), observed according to the GPCC (land) and GPCP (ocean) datasets; (c) as (a), for the cut-cell model LMZ; (d) as (a), for the terrain-following model CLM in its current version. For (c) and (d), the same colour scheme as for (b) was used. The lines and the numbers 1 to 4 in (b) denote the areas used to calculate the precipitation biases. The forecasted accumulated precipitation for the whole January 1989 using the terrain-following model LM; (b) as (a o the GPCC (land) and GPCP (ocean) dataset; (c) as (a), for the cut-cell model LMZ; (d) as (a), for the terrain-follow current version. For (c) and (d), the same colour scheme as for (b) was used. The lines and the numbers 1 to 4 in (b) Fig. 4. 4 Conclusions (a) The forecasted accumulated precipitation for the whole January 1989 using the terrain-following model LM; (b) as (a), observed according to the GPCC (land) and GPCP (ocean) datasets; (c) as (a), for the cut-cell model LMZ; (d) as (a), for the terrain-following model CLM in its current version. For (c) and (d), the same colour scheme as for (b) was used. The lines and the numbers 1 to 4 in (b) denote the areas used to calculate the precipitation biases. Table 2. Temperature anomaly correlations for LMZ and LM for selected locations. scheme (see, e.g. Hollweg et al., 2008). These desirable im- provements are not yet implemented in the LMZ. In spite of this, the LMZ shows a better localisation of precipitation, even though in other aspects the CLM model gives better pre- cipitation forecasts. Table 2. Temperature anomaly correlations for LMZ and LM for selected locations. City name location Corr (LMZ) Corr (LM) Beijing 40◦N, 117◦E 0.80 0.16 Baoutu 41◦N, 110◦E 0.83 0.60 Yinchouan 38◦N, 105◦E 0.90 0.24 Gan Ze 33◦N, 100◦E 0.78 0.14 Xian 34◦N, 108◦E 0.55 0.34 Acknowledgements. The ﬁrst author thanks NCAR for support of this work. The forecasts were done on the NCAR blueﬁre computer and visits of the ﬁrst author to Boulder were ﬁnanced by NCAR. ERA-interim data were provided by the European Centre for Medium-Range Weather Forecasts. the LM having large differences to the analysis. As shown for shorter forecasts in Stal06, the vertical velocities and precipi- tation are more realistic for the LMZ model, when compared to the control model LM. The small improvements of tem- perature and wind forecasts reported in Stal06 become more substantial after ﬁve days. Over large areas the temperatures and winds are improved, when using the analysed ﬁelds as veriﬁcation. 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https://openalex.org/W4379519882	OpenAlex	Open Science	CC-By-SA	2,023	Perubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0	Rosa Indah	Indonesian	Spoken	3,077	7,186	Email: Korespondensi penulis: rosaindahs21@gmail.com Abstrak, Perkembangan zaman mempengaruhi mempengaruhi perkembangan perbankan di Indonesia. Penelitian ini bertujuan untuk menganalisis tranformasi bank dari bank konvensional menjadi bank digital. Jenis penelitian yang digunakan adalah deskriptif kualitatif menggunakan data sekunder dari sumber terpercaya. Hasil dari penelitian ini menunjukkan bahwa transformasi bank konvensional menjadi bank digital memberikan banyak keuntungan seperti akses layanan tanpa batas, tarif yang semakin efisien, ragam fitur beyond banking, keamanan, kontrol finansial dan kemudahan dalam pengajuan kredit. Bank digital juga memberikan kemudahan bagi pengguna dalam melakukan berbagai aktivitas perbankan, mulai dari pembukaan akun, transfer dana, penempatan deposito, hingga penutupan akun, semuanya dapat dilakukan melalui perangkat elektronik seperti smartphone tanpa perlu mengunjungi kantor cabang secara fisik. ata Kunci : Bank Kovensional, Bank Digital, Tranformasi, Revolusi industri 4.0 Kata Kunci : Bank Kovensional, Bank Digital, Tranformasi, Revolusi industri 4.0 Received April 30, 2023; Revised Mei 02, 2023; Juni 01, 2023 Corresponding author, e-mail address Perubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0 Rosa Indah Universitas 17 Agustus 1945 Surabaya Email: rosaindahs21@gmail.com Ummukifdiyah Rohmatul Huda Universitas 17 Agustus 1945 Surabaya Email: awdiyahawdiyah@gmail.com Anjar Puji Asmeru Universitas 17 Agustus 1945 Surabaya Email: anjarpuji7@gmail.com Muhammad Yasin Universitas 17 Agustus 1945 Surabaya Email: yasin@untag-sby.ac.id p , ; Corresponding author, e-mail address 1. PENDAHULUAN Bank memainkan peran penting dalam membangun ekonomi nasional dan memiliki posisi strategis dalam menggerakkan roda perekonomian. Lembaga perbankan merupakan inti dari sistem keuangan di setiap negara dan bertujuan utama untuk menghimpun dan menyalurkan dana masyarakat serta menunjang pelaksanaan pembangunan nasional. Fungsi utama bank adalah sebagai penghimpun dan penyalur dana masyarakat serta memberikan pembiayaan dan jasa dalam lalu lintas pembayaran dan peredaran uang. Lembaga keuangan mencakup semua entitas yang melalui kegiatan di bidang keuangan, menarik dan menyalurkan uang ke masyarakat. Saat ini, di Indonesia, umumnya digunakan bank konvensional. Bank konvensional adalah bank yang menjalankan aktivitasnya secara tradisional, mengacu pada kesepakatan baik dalam konteks nasional maupun internasional, serta berdasarkan hukum formal negara. Student Research Journal Vol.1, No.3 Juni 2023 DOI: https://doi.org/10.55606/srjyappi.v1i3 Meskipun demikian, bank konvensional memiliki tiga kelemahan, yakni 1) masalah manajemen yang ditandai oleh ketidak sesuaian dalam penyaluran kredit, campur tangan pemilik yang berlebihan, dan kurangnya profesionalisme manajer; 2) kredit bermasalah akibat ketidak patuhan dalam prosedur pemberian kredit dan akumulasi pemberian kredit kepada kelompok tertentu; 3) praktik-praktik curang seperti bank dalam bank dan transaksi fiktif (Kasmir, 2010). Menghadapi tantangan tersebut, Bank Indonesia harus siap mengikuti perubahan zaman dan beralih ke digitalisasi perbankan dalam era revolusi industri 4.0. Perkembangan zaman telah menghadirkan Revolusi Industri 4.0, yang juga dikenal sebagai sistem cyber-physical, yang menekankan pada otomatisasi dan kolaborasi antar teknologi. Dampak revolusi ini terasa luas, termasuk pada perkembangan internet yang menjadi kebutuhan penting dalam kehidupan sehari-hari manusia. Salah satu sektor industri yang mengalami transformasi adalah industri keuangan di Indonesia, terutama sektor perbankan. Industri keuangan Indonesia mengalami kemajuan yang sangat pesat dalam bidang teknologi serta perbankan digitalnya. Perkembangan ini ditandai dengan kemunculan berbagai sistem pembayaran instan dan mudah, seperti layanan perbankan melalui internet (internet banking), perbankan melalui ponsel (mobile banking), mesin ATM, e-money, serta berbagai fitur lainnya.. Perkembangan yang pesat dalam industri fintech di Indonesia telah mengubah cara masyarakat melakukan transaksi keuangan. Hal ini penting untuk meningkatkan keandalan dan responsivitas, keamanan, serta efisiensi operasional dan layanan perbankan secara keseluruhan. Seluruh proses ini mengarah pada transformasi dari bank konvensional menjadi bank digital yang lebih inovatif dan memberikan hasil yang signifikan. 2.1 Perbankan Menurut Undang-Undang Perbankan No. 7 Tahun 1992 tentang Perbankan yang telah diubah menjadi Undang-Undang No. 10 Tahun 1998, mengartikan bank sebagai perusahaan yang bergerak di bidang jasa dan memiliki kegiatan pokok dengan tiga fungsi pokok, sebagai berikut: menerima penyimpanan dana masyarakat dalam berbagai bentuk, menyalurkan dana tersebut dalam bentuk kredit kepada masyarakat untuk mengembangkan usaha, dan melaksanakan berbagai jasa dalam kegiatan perdagangan dan pembayaran dalam negeri maupun luar negeri serta berbagai jasa lainnya di bidang keuangan (IBI, 2013). Bank dalam melaksanakan kegiatannya mempunyai kebebasan untuk menentukan produk dan jasanya. Kegiatan menghimpun dana masyarakat dilakukan melalui penjualan produk dana dalam bentuk Giro, Tabungan, Deposito dan Sertifikat Deposito. Kegiatan menyalurkan dana dilakukan melalui penyaluran kredit dalam berbagai bentuk seperti Kredit Investasi, Kredit Modal Kerja, Kredit Konsumtif, Kredit tanpa jaminan, dll. 169 SRJ - VOLUME 1, NO. 3, JUNI 2023 169 erubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0 Perubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0 Kegiatan jasa perbankan merupakan kegiatan yang dilakukan perbankan untuk memperlancar terjadinya transaksi perdagangan, memperlancar peredaran uang, dan memberikan jaminan kepada nasabahnya.Beberapa jasa yang ditawarkan oleh perbankan antara lain transfer, inkaso, kliring, BI-RTGS, bank draft, traveller’s cheque, letter of credit, bank garansi, safe deposit box, jual beli valuta asing dan electronic banking. Elektronic banking adalah salah satu produk jasa perbankan yang saat ini paling diminati oleh masyarakat seiring dengan perkembangan teknologi dan kebutuhan masyarakat yang semakin beragam. Elektronic banking atau dikenal dengan e-banking merupakan sistem yang memungkinkan nasabah bank baik individu ataupun bisnis untuk mengakses rekening, melakukan transaksi bisnis, atau mendapatkan informasi produk dan jasa bank melalui jaringan pribadi atau publik termasuk internet Penyelenggaraan e-banking merupakan penerapan atau aplikasi teknologi informasi yang terus berkembang dan dimanfaatkan untuk menjawab keinginan nasabah perbankan yang menginginkan servis cepat, aman, nyaman, murah, dan tersedia setiap saat 24 jam/hari dan dapat diakses dari mana saja (Rosa, 2016). 2.2 Digitalisasi Perbankan Penelitian yang dilakukan oleh Kholis (2018) menunjukkan bahwa industri perbankan dalam memberikan layanan harus beradaptasi dengan perkembangan teknologi untuk memberikan kemudahan yang diinginkan oleh nasabah terhadap layanan perbankan. Perkembangan Financial Technology atau yang dikenal dengan istilah FinTech sejak tahun 2015 membuat perbankan harus aware dengan perkembangannya jika tidak ingin ditinggalkan nasabahnya yang dapat beralih ke lembaga keuangan lain. Tidak hanya di Indonesia, digitalisasi perbankan merupakan sesuatu yang tidak terhindarkan lagi di berbagai belahan dunia. Anthony Rahul Golden S (2017) dalam tulisannya mengenai digitalisasi dalam sektor perbankan di India menyimpulkan “Due to the adoption of this digitalization, the banking sectors in India face some remarkable changes as well as hurdles. As we are in the digital era, it is not possible to avoid the growth and services or digital banking. Everyone uses the modern mobile device, called Smartphone, which is used to access the digital banking services to anyone at anywhere and at any time. Thus, digital banking is in the place of inevitable today”. Jadi dapat disimpulkan bahwa digitalisasi perbankan saat ini bukan merupakan sebuah pilihan, melainkan telah menjadi suatu keharusan bila bank ingin terus bersaing memperebutkan pangsa pasar yang tersedia. Digitalisasi perbankan tentunya berdampak positif bagi bank maupun nasabahnya. Transaksi yang tidak lagi dibatasi tempat dan waktu menjadi keuntungan yang signifikan bagi nasabah, sementara bagi perbankan kenaikan pendapatan yang berasal dari fee based income dan penurunan biaya tenaga kerja antara lain merupakan dampak positif yang dirasakan. Saviour dan Bornwell (2018) dalam rekomendasinya menuliskan “The banks make information on the benefits, costs, how the use etc available 170 SRJ - VOLUME 1, NO. 3, JUNI 2023 170 Student Research Journal Vol.1, No.3 Juni 2023 DOI: https://doi.org/10.55606/srjyappi.v1i3 to the all customers and merchants. The use of e-banking has been found to be very beneficial to both the customer and the bank because the customer believes the service reduces transaction time; it is convenient and reduces the amount of cash to be carried in order to do a transaction which reduces the risk of losing cash. To the Bank, the use of e- banking has helped to reduce cost of handling the transactions including reduced cost of back room staff.” Hal ini menekankan bahwa dampak e-banking terhadap tenaga kerja tidak bisa dihindari. 2.2 Digitalisasi Perbankan Mukhtar (2015) dalam penelitiannya mengenai persepsi nasabah di United Kingdom terhadap internet banking memberikan pertanyaan apakah responden menggunakan internet banking dan jawaban yang didapatkannya adalah bahwa “most of the participants (53%) did not use internet banking. However, there were 47% participants who had experience of using internet banking services. Many earlier studies have revealed that internet banking is still not completely adopted by the customers. In fact, many earlier studies noted that users of the traditional banking system are more than internet banking users”. Dalam penelitian lain di India, S.V. Mohana Sujana (2018) juga menyampaikan bahwa “Only 64% of the population are aware of the digital banking services and make use of it”. Jadi masih terdapat segmen nasabah yang tetap lebih menyukai dan merasa nyaman menggunakan layanan perbankan tradisional, terutama golongan nasabah yang berusia lebih tua. Oleh karena itu bank tetap memperhatikan keseimbangan layanan perbankan tradisionalnya mengingat potensi pasarnya yang masih cukup besar. Digitalisasi perbankan juga memiliki beberapa tantangan dalam perkembangannya.S.V. Mohana Sujana (2018) menuliskan beberapa kekurangan dari digitalisasi perbankan yaitu Personal relationship between the bank officials and the customers has been minimized due to access from one place. As everything has been mobilised the security in protecting transaction has been reduced in such a case issues regarding transactions have been increased. There is an high risk in making transactions because identification of theft of encrypted software.Tentunya hal-hal tersebut merupakan faktor-faktor yang harus diperhatikan dan menjadi bahan pertimbangan dunia perbankan dalam mengembangkan dan memodernisasi layanannya. Dalam penelitian di GCB Bank Ghana, Martin Otu Offei, dkk., (2016) menyampaikan bahwa tingginya biaya internet di Ghana menjadi salah satu hambatan bagi nasabah dalam menggunakan layanan internet banking. Hasil survey menyatakan “With regard to problems associated with internet banking, result indicate that the major challenge given by the respondents was that of high cost of internet service charges, representing 83%. This was followed by high cost of maintaining internet facility and high cost of getting reliable internet connectivity, representing 10% and 7% respectively”. 3. METODE PENELITIAN 171 SRJ - VOLUME 1, NO. 3, JUNI 2023 Perubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0 Penelitian ini dilakukan dengan cara mencari mengumpulkan sumber informasi jurnal dan internet yang berkaitan dengan bank konvensional dan bank digital. Sumber-sumber tersebut kemudian dianalisis secara mendalam (Hasan , Dzakiyyah , Kumalasari , Safira, & Aini, 2021). Pencarian literatur bertujuan untuk mendapatkan informasi yang sesuai dan melakukan evalusasi pada sumber yang diacu. Data yang digunakan dalam penelitian ini adalah data sekunder, yang kemudian dianalisis menggunakan pendekatan deskriptif kualitatif. Penelitian ini dilakukan dengan mencari literatur dan sumber informasi di internet dan jurnal tranformasi bank konvensional menjadi bank digital yang kemudian dianalisis lebih lanjut (Hasan , Dzakiyyah , Kumalasari , Safira, & Aini, 2021). Tujuan utama dari pencarian literatur ini adalah untuk memperoleh informasi yang relevan dan untuk mengetahui sejauh mana informasi tersebut terkait dengan orang lain, apakah didukung atau tidak. Data dalam penelitian ini adalah data sekunder yang dianalisis menggunakan dekriptif kualitatif. 4. HASIL DAN PEMBAHASAN Transformasi perbankan menjadi perbankan digital adalah proses di mana lembaga keuangan seperti bank mengadopsi teknologi digital untuk meningkatkan kualitas layanan dan efisiensi operasional bisnisnya (Winasis, Riyanto , & Ariyanto , 2020). Bertransformasi menjadi bank digital melibatkan pemanfaatan teknologi informasi untuk mengubah cara bank berinteraksi dengan nasabah, melakukan transaksi keuangan, serta mengelola risiko. Tujuan transformasi digital perbankan adalah untuk membuat layanan perbankan menjadi lebih mudah diakses, lebih cepat, lebih aman, dan lebih efisien. Beberapa contoh dari transformasi digital perbankan seperti penyediaan layanan perbankan online dan mobile banking dalam satu platform aplikasi yang dapat diakses melalui smartphone, serta integrasi data dan konektivitas jaringan antara kantor pusat dan kantor cabang guna meningkatkan kepuasan pelanggan. Dalam era serba digital, layanan phone banking juga menjadi salah satu perkembangan teknologi call center yang dapat diakses selama 24 jam. Bank digital memberikan berbagai kelebihan, antara lain kemudahan dalam melakukan transaksi di mana pun dan kapan pun, transparansi dalam transaksi, serta biaya yang lebih rendah, serta meningkatkan kemampuan bank untuk menciptakan ekosistem yang bekerjasama dengan penyedia e- commerce guna mencapai pengguna superApp. Bank digital adalah bank yang menyediakan layanan perbankan melalui sarana elektronik atau secara online. Bank digital memberikan kelebihan berupa akses mudah dan cepat ke layanan keuangan, termasuk melakukan transaksi, transfer dana, pembayaran tagihan, dan pembelian produk secara online. Selain itu, pengguna juga dapat dengan mudah melacak dan mengelola riwayat transaksi mereka, serta memantau perkembangan keuangan mereka melalui fitur pelaporan dan analisis yang disediakan SRJ - VOLUME 1, NO. 3, JUNI 2023 172 Student Research Journal Vol.1, No.3 Juni 2023 J e-ISSN: 2964-3252; p-ISSN: 2964-3260, Hal 168-175 DOI: https://doi.org/10.55606/srjyappi.v1i3 p g jy pp oleh platform digital tersebut. Transaksi keuangan digital terus tumbuh tinggi seiring dengan hadirnya penggunaan platform e-commerce (Marsudi & Widjaja , 2019). Perkembangan teknologi keuangan yang pesat menunjukkan bahwa teknologi baru dapat memainkan peran strategis, terutama di Indonesia, dengan memberikan akses mudah dan cepat ke layanan keuangan tanpa harus bertemu secara fisik.. Ketersediaan fasilitas ini juga didukung dengan sangat baik oleh setiap individu maupun pelaku bisnis di zaman milenial seperti saat ini. Dengan adanya teknologi dan digital keuangan seperti saat ini maka motivasi pelanggan untuk berbelanja akan lebih loyal dan daya beli masyarakat pun dapat meningkat secara pesat (Gouveia, Perun , & Daradkeh, 2020). Berdasarkan teori yang diacu, bank yang telah melakukan digitalisasi adalah Bank DBS milik Singapura. Bank miliki Singapura membantu nasabah dengan baik karena bank yang sederhana, nyaman, dan tampa hambatan (Gupta, 2021). 4. HASIL DAN PEMBAHASAN Keuntungan menggunakan bank digital antara lain fleksibel dan praktis karena semua layanan keuangan dapat dilakukan secara online tanpa harus mendatangi kantor cabang bank. Selain itu, tarif yang semakin efisien, ragam fitur beyond banking, keamanan, kontrol finansial dan personalisasi juga menjadi manfaat dari bank digital. Bank digital juga dapat membantu meningkatkan inklusi keuangan dan mempermudah akses perbankan bagi masyarakat. Namun, bank digital juga menghadapi tantangan seperti manajemen risiko yang memadai, pengembangan ekosistem keuangan digital, dan sistem keamanan yang efektif. Sumber: (Barquin & Vinayak , 2015) Gambar 1. Struktur Organisasi Baru Untuk Meningkatkan Rencana Transformasi Digital Sumber: (Barquin & Vinayak , 2015) Gambar 1. Struktur Organisasi Baru Untuk Meningkatkan Rencana Transformasi Digital Transformasi digital adalah proses penggunaan teknologi informasi dan komunikasi untuk melakukan perubahan pada proses kegiatan masyarakat. Tranformasi digital dalam bisnis merujuk pada bagaimana teknologi yang mendukung revolusi bisnis menggunakan internet. Transformasi digital dapat mengubah Indonesia, memunculkan potensi ekonomi baru dan tantangan talenta digital. Teknologi menjadi alat yang digunakan manusia untuk menunjang pemenuhan kebutuhan serta permintaan. 173 SRJ - VOLUME 1, NO. 3, JUNI 2023 173 erubahan Bank Di Indonesia Dari Bank Konvensial Menjadi Bank Digitalisasi Di Era Evaluasi Industri 4.0 Transformasi digital juga dapat meningkatkan aksesibilitas dan konektivitas, memperluas akses terhadap infrastruktur TIK seperti internet, telepon seluler, dan jaringan. Transformasi digital penting untuk meningkatkan ketangkasan dan efisiensi operasional dalam bisnis serta membuka nilai dalam proses bisnis dan melepaskannya kembali ke pelanggan. Transformasi digital juga dapat membantu organisasi untuk menjadi lebih efektif dan efisien dalam upaya meningkatkan pengalaman pelanggan. Transformasi digital adalah proses integrasi teknologi digital ke dalam semua area bisnis, yang mengakibatkan perubahan mendasar dalam cara operasional dan memberikan nilai kepada pelanggan. Ini juga merupakan perubahan budaya yang membutuhkan organisasi untuk terus-menerus menggoyang status quo, bereksperimen, dan nyaman dengan teknologi baru. Transformasi digital sangat penting bagi semua bisnis, mulai dari skala kecil hingga perusahaan besar, karena hal ini diperlukan untuk tetap kompetitif dan relevan dalam dunia yang semakin digital. Proses ini melibatkan penggunaan teknologi digital untuk mengubah proses bisnis dan layanan tradisional yang sudah ada atau menciptakan yang baru untuk memenuhi harapan pasar dan pelanggan yang terus berkembang. Tujuan yang mengarahkan implementasi dapat bervariasi dari perusahaan ke perusahaan tetapi semua inisiatif transformasi digital bertujuan untuk meningkatkan hasil bisnis dengan mengadopsi teknologi digital modern. Student Research Journal Vol.1, No.3 Juni 2023 e-ISSN: 2964-3252; p-ISSN: 2964-3260, Hal 168-175 Barquin , S., & Vinayak . (2015). Digital Banking in Asia : What do consumers really want?McKinsey & Company , 1-12. Gouveia, L. B., Perun , M., & Daradkeh, Y. I. (2020). DIGITAL TRANSFORMATION AND CUSTIMERS SERVICES: the BANKING REVOLUTION. INTERNASIOLAN JOURLAN OF OPEN INFORMATION TECHNOLOGIES , 8(7):124-128 . 5. KESIMPULAN DAN SARAN Berdasarkan hasil penelitian yang dilakukan, diketahu bahwa Bank digital adalah inovasi teknologi layanan perbankan yang memungkinkan pengguna untuk mengakses segala kebutuhan finansial melalui saluran elektronik tanpa harus datang ke kantor fisik. Bank digital memiliki beberapa keunggulan, seperti meningkatkan keamanan dan kenyamanan transaksi, memenuhi standar yang ditetapkan OJK, dan bersedia ikut andil untuk kemajuan ekosistem keuangan digital dan pemenuhan hak masyarakat Indonesia dalam mengakses perbankan. Manfaat lain dari bank digital adalah akses layanan tanpa batas, tarif yang semakin efisien, ragam fitur beyond banking, keamanan, kontrol finansial dan kemudahan dalam pengajuan kredit. Bank digital juga memberikan kemudahan bagi pengguna dalam melakukan berbagai aktivitas perbankan, mulai dari pembukaan akun, transfer dana, penempatan deposito, hingga penutupan akun, semuanya dapat dilakukan melalui perangkat elektronik seperti smartphone tanpa perlu mengunjungi kantor cabang secara fisik. Perbedaan antara bank digital dengan layanan digital banking dari bank konvensional adalah bank digital umumnya tidak memiliki kantor fisik (selain kantor pusat) atau dapat menggunakan kantor fisik yang terbatas. SRJ - VOLUME 1, NO. 3, JUNI 2023 174 Student Research Journal Vol.1, No.3 Juni 2023 e-ISSN: 2964-3252; p-ISSN: 2964-3260, Hal 168-175 DOI: https://doi.org/10.55606/srjyappi.v1i3 DAFTAR PUSTAKA Barquin , S., & Vinayak . (2015). Digital Banking in Asia : What do consumers really want?McKinsey & Company , 1-12. Gouveia, L. B., Perun , M., & Daradkeh, Y. I. (2020). DIGITAL TRANSFORMATION AND CUSTIMERS SERVICES: the BANKING REVOLUTION. INTERNASIOLAN JOURLAN OF OPEN INFORMATION TECHNOLOGIES , 8(7):124-128 . Gupta, P. (2021). DBS BANK OF SINGAPORE . SINGAPURA. Hasan , M., Dzakiyyah , A., Kumalasari , D. A., Safira, N., & Aini, S. N. (2021). TRANSFORMASI DIGITAL UMKM SEKTOR KULINER DI KELUARAHAN JATINEGARA JAKARTA TIMUR. JURNAL BISNIS DAN KEWIRASUHAAN, 17(2):135-150. Kasmir. (2010). Manajemen Perbankan. Edisi Revisi ke Sembilan . Jakarta : PT Raja Grafindo Persada. Marsudi , A. S., & Widjaja , Y. (2019). INDUSTRI 4.0 DAN DAMPAKNYA TERHADAP FINANCIAL TECHNOLOGU SERTA KESIAPAN TENAGA KERJA DI INDONEISA . IKRA-ITG EKONOMIKA, 2(2):1-10. Winasis, S., Riyanto , S., & Ariyanto , E. (2020). Digital Transformation in Indonesian Banking Industry: Impact on Employee Engagement. International Journal of Innovation, Creativity and Change. International Journal of Innovation, Gupta, P. (2021). DBS BANK OF SINGAPORE . SINGAPURA. Hasan , M., Dzakiyyah , A., Kumalasari , D. A., Safira, N., & Aini, S. N. (2021). TRANSFORMASI DIGITAL UMKM SEKTOR KULINER DI KELUARAHAN JATINEGARA JAKARTA TIMUR. JURNAL BISNIS DAN KEWIRASUHAAN, 17(2):135-150. Kasmir. (2010). Manajemen Perbankan. Edisi Revisi ke Sembilan . Jakarta : PT Raja Grafindo Persada. Marsudi , A. S., & Widjaja , Y. (2019). INDUSTRI 4.0 DAN DAMPAKNYA TERHADAP FINANCIAL TECHNOLOGU SERTA KESIAPAN TENAGA KERJA DI INDONEISA . IKRA-ITG EKONOMIKA, 2(2):1-10. Winasis, S., Riyanto , S., & Ariyanto , E. (2020). Digital Transformation in Indonesian Banking Industry: Impact on Employee Engagement. International Journal of Innovation, Creativity and Change. International Journal of Innovation, Creativity and Change , 1(2):12-20. 5 SRJ - VOLUME 1, NO. 3, JUNI 2023 SRJ - VOLUME 1, NO. 3, JUNI 2023	28,816
66/dumas.ccsd.cnrs.fr-dumas-00592362-document.txt_3	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	6,951	10,054	Cette mesure reste partielle, car les clients n'expriment pas toujours leur mécontentement, certains d'ailleurs quittent leurs fournisseurs sans leur adresser de signes visibles de mécontentement. Les réclamations peuvent donner lieu à des actions correctives (modification d'un prix erroné, échange d'un produit défectueux, modification du stock) ou a des modifications du système qualité : création d'une procédure (gestion des périmés), création d'un projet qualité (extension du parc d'aérosol, de fauteuils roulant) L'analyse faite par la CCI se fait en deux temps, par le passage de deux « clients mystères » qui mesurent l'approche et l'accueil du client. Leur analyse se base sur des aspects « perceptuels », sur leur ressenti en tant que client dans notre officine. Les résultats obtenus lors de ces visites ne nous ont pas encore été communiqués. Principe 2 - Le leadership ou l'engagement du titulaire Les dirigeants établissent la finalité et les orientations de l'organisme. Il convient qu'ils créent et maintiennent un environnement interne dans lequel les personnes peuvent pleinement s'impliquer dans la définition et la réalisation des objectifs de l'organisme. [43] L'entreprise doit prendre en compte des besoins de toutes les parties intéressées notamment les clients, les employés, les fournisseurs, les financiers, les collectivités locales et la société dans son ensemble. L'entreprise présentée dans ce document s'implique à travers son titulaire dans cette démarche, qui ne portait pas le nom de qualité auparavant, mais qui existe depuis de nombreuses années. Plusieurs « modes d'emploi », « protocoles », ou « modes opératoires » existaient déjà avant la mise en place d'une réelle démarche qualité. Cette politique n'a fait qu'évoluer en fonction de la société, de ses besoins et de ses attentes. Le chef d'entreprise s'est largement engagé dans cette démarche que ce soit au sein de notre officine ou au sein du groupement dont nous faisons partie pour promouvoir la politique qualité dans de nombreuses officines. Si l'engagement du titulaire est réel, il doit également mettre en oeuvre un environnement et les moyens adéquats pour satisfaire les objectifs fixés par l'entreprise. Comme nous l'avons vu, une entreprise se caractérise par ses employés. La qualité du travail qu'ils vont fournir va déterminer la satisfaction des clients de l'entreprise. Cette qualité même de travail doit pouvoir être maintenue constante, voire améliorée. Pour se faire il serait 95 nécessaire de fixer des objectifs ainsi que les moyens pour parvenir aux objectifs. Si ces moyens ne sont pas adaptés, cela peut engendrer une situation de stress pour les employés. Il peut être mis en place des objectifs personnels ou communs objectiv à échéance par les résultats obtenus et éventuellement récompensés pour stimuler la motivation des membres du personnel. Principe 3 - L'implication du personnel Les personnes à tous niveaux sont l'essence même d'un organisme et une totale implication de leur part permet d'utiliser leurs aptitudes au profit de l'organisme. [43] Le personnel doit être motivé, impliqué et engagé pour l'organisme. Il doit comprendre l'importance de sa contribution et son rôle dans l'organisme, en acceptant d'être responsabilisé et d'assumer sa part de responsabilité à résoudre les problèmes. Le personnel participe et contribue à l'amélioration continue. Lors de ce travail sur la dispensation sur prescription utilisant la méthode Delphi, l'ensemble de l'équipe a été mise à contribution. Chacun a pu comprendre pourquoi et comment sont réalisées les procédures ainsi que l'ensemble des documents du système qualité. Chacun a eu la possibilité de se poser des questions sur l'assurance qualité et sur le service au patient. Et ainsi s'interroger sur leur manière de procéder dans leurs activités quotidiennes. La participation des membres de l'équipe à la rédaction de ce processus et des procédures améliore l'engagement dans la démarche. Les documents du système qualité crées par l'ensemble du personnel sont mieux connus, mieux perçus et donc mieux suivis. L'ensemble de l'équipe a très bien perçu le travail demandé et a été très satisfait de pouvoir y participer. Lors de la création d'un grand nombre de procédures du système qualité de la pharmacie (réception de commande, chaine du froid, gestion de la location, 96 procédure de tiers-payant) l'équipe n'a eu qu'un rôle consultatif a postériori, ce qui à la suite de ce travail n'apparait pas comme la solution la plus adaptée. Il est indispensable que pour le bon fonctionnement et l'évolution du système qualité, l'ensemble du personnel continue d'intervenir de manière plus importante dans la rédaction des procédures. Pour se sentir concerné chacun des membres du personnel doit intervenir sur le choix des procédures nécessaires ainsi que sur la rédaction et la mise en place de celles-ci. Grâce à cette méthode chacun des membres de l'entreprise sait pourquoi il existe une procédure (ils l'ont choisie), sait ce que comporte la procédure (ils ont participé à la rédaction et la en place), et pourront donc chercher à l'améliorer avec le temps et l'expérience. L'implication du personnel dépend de la motivation du personnel, cette motivation dépend de la satisfaction du personnel. S. Araye dans son récent travail [9] s'interroge sur les conséquences de la mise en place de l'assurance qualité et plus précisément sur la certification ISO 9001 :2000 sur le personnel au sein d'une agence d'un grossiste répartiteur. Il en conclut que « si le personnel n'est pas satisfait il peut y avoir apparition de résistance vis-à-vis du changement, néfaste au projet mis en oeuvre mais également à la qualité de travail de l'entreprise et donc source de diminution de la satisfaction client. Principe 4 - L'approche processus Schématiser les activités sous forme d'un « processus » consiste à définir pour chaque activité les objectifs à atteindre en planifiant si besoin les différentes étapes de la réalisation, ce qui permet de mieux gérer ces activités pour obtenir d'une manière efficace le résultat attendu. Ceci implique une définition systématique des activités nécessaires pour obtenir un résultat désiré ; d'établir des responsabilités claires pour la gestion des activités clés ainsi que les ressources, les méthodes et les matériels qui amélioreront ces activités. Comme présenté dans ce document les grandes activités de l'entreprise sont désormais décrites par des processus qui indiquent quels sont les points importants à réaliser pour mener à bien ces activités. Processus de dispensation, processus de fabrication, prochainement processus d'accueil client et de maitrise des approvisionnements. L'établissement de ces processus a permis de définir les grandes activités de l'officine sous la forme de schémas simples qui ont permis de remplacer les anciennes procédures qui étaient très rédactionnelles et donc très lourdes d'utilisation. Une approche simplifiée de la démarche qui met en avant des objectifs clairs : « satisfaction client », « maitrise de la chaine du froid », « fabrication et traçabilité des préparations magistrales » permettant une meilleure compréhension et donc implication du personnel dans cette politique. Principe 5 - Le management par approche système Comprendre que l'ensemble des différents processus forme un système stable orienté vers les objectifs finaux permettra plus facilement à l'organisme d'atteindre ces objectifs. [43]. Il est nécessaire de structurer le système pour atteindre les objectifs de l'organisme de 98 la façon la plus efficace et efficiente, de comprendre les interdépendances entre les processus du système et d'améliorer continuellement le système par le biais de mesures et d'évaluations. La lecture de la cartographie aide à mieux comprendre que chaque activité est liée aux autres, certaines sont même fortement dépendantes les unes des autres. Une dispensation conforme à notre référentiel nécessite une bonne gestion des approvisionnements en médicaments, une gestion de stock approprié (pas de périmés, les produits sont situés à la bonne place), un personnel formé, des outils adaptés (logiciel informatique) si un des systèmes est défaillant c'est le résultat final qui n'est pas à la hauteur de nos objectifs. Les objectifs finaux de l'entreprise sont donc dépendants des objectifs de chacun des processus et activités réalisées. Pour mesurer ces objectifs, chacune des activités doit être soumise à des indicateurs de fonctionnement. A l'heure actuelle la mise en place d'indicateur de mesure est faible au sein de notre entreprise car lourde à mettre en place. Pour l'instant il existe un recueil des erreurs de stock pour permettre une correction rapide mais pas d'analyse de ces erreurs, il a été également mis en place un recueil des erreurs de délivrance au sein de l'officine. Il s'agit de noter les erreurs effectuées par les membres de l'équipe lors de la dispensation, en notant le nom de la personne concernée, le nom du client ainsi que la description de l'erreur. Cette mesure permet d'identifier les sources d'erreurs au moment de la délivrance, d'y apporter une correction immédiate ou d'envisager une action corrective plus importante si l'erreur se répète. Malheureusement erreurs ici notées semblent pour le plus souvent être des erreurs humaines (étourderie, précipitation, mauvaise lecture de la prescription,) et le recueil de ces erreurs apparait au sein de l'équipe comme une certaine « dénonciation » qui de plus n'entraine que très rarement des actions correctives importantes. 99 Il existe d'autres mesures qui pourraient être réalisées, par exemple le recueil et l'analyse des produits manquants au moment de la délivrance, afin de comprendre pourquoi certains produits sont régulièrement manquants et le corriger. La satisfaction du client serait ici une conséquence directe de la meilleure gestion de notre stock. Principe 6 - L'amélioration continue L'amélioration continue de la performance globale doit être un objectif permanent de l'organisme. Il est important d'assurer une « veille » permanente portant sur les différentes évolutions : besoins, produits, marchés, technologie, méthodes, compétences, voire une anticipation de ces évolutions (projets). [43] Améliorations dans nos méthodes mais également améliorations dans nos offres de produits et services. Développement du maintien à domicile (l'officine a agrandi dernièrement son parc de matériel de location avec des fauteuils roulants en location), développement des produits composés de plantes et de produits issus de l'agriculture biologique. L'entreprise assure cette veille sur le marché de la santé grâce aux informations relayées par nos laboratoires fournisseurs, notre groupement, la presse spécialisé et non spécialisée. Pour mettre en place l'amélioration continue de notre politique qualité, il est nécessaire d'assurer la formation du personnel aux méthodes et outils d'amélioration continue pour que l'amélioration des produits, processus et systèmes devienne un objectif de chaque individu dans l'organisme. Mais il apparait difficile au sein d'une équipe d'émettre des critiques sur le travail réalisé. Dans notre activité quotidienne on ne s'aperçoit pas des erreurs que nous commettons. Il 100 est difficile de critiquer une méthode que nous pensons être la bonne et que nous utilisons depuis de nombreuses années. Il est pourtant indispensable de se remettre en cause régulièrement pour envisager les différentes améliorations à apporter, ces améliorations ne sont pas obligatoirement des modifications du système qualité. Elles peuvent être inhérentes aux personnes (non-conformité aux règles et procédures mise en place, erreurs humaines) C'est dans ce cadre que chacun doit évoluer dans démarche d'amélioration personnelle puis collective. Pour nous aider dans cette tâche il est important d'avoir des avis extérieurs de notre travail. C'est pour cette raison que nous accordons une grande importance aux remarques de nos clients mais aussi à la venue de clients-mystère par exemple. On se doit d'inciter les membres de l'équipe à la critique qui permettrait d'améliorer nos méthodes de travail. Ces remarques et critiques sur l'organisation de la pharmacie, sur nos méthodes de travail sont recueillies lors des « réunions qualité » qui se déroulent une fois par mois. Afin d'engager une discussion sur une possible voie d'amélioration. Il est indispensable, comme le prévoit la norme ISO 9001 : 2008, d'effectuer des audits internes à intervalles réguliers. Ces deux évaluations nous ont donné des résultats probants dans notre démarche de développement de l'assurance qualité, mais aujourd'hui avec l'évolution de notre système de management, il nous faudrait réaliser un audit plus adapté aux exigences de la Norme internationale et aux exigences du système de management de la qualité établies par l'entreprise. Or cette méthode d'évaluation de nos pratiques quotidiennes semble difficile à mettre en place pour 101 des personnes non spécialisées dans le domaine de l'Assurance Qualité, d'autant plus que la norme précise que le choix des auditeurs et la réalisation des audits doivent assurer l'objectivité et l'impartialité du processus d'audit et que les auditeurs ne doivent pas auditer leur propre travail. Ce qui semble difficile à réaliser dans une entreprise comme une pharmacie d'officine. Principe 7 - L'approche factuelle pour la prise de décision Les décisions efficaces se fondent sur l'analyse de données et d'informations qui doivent être exactes, fiables et accessibles à ceux qui en ont besoin. Les prises de décisions et les actions doivent être fondées sur une analyse factuelle, équilibrée par l'expérience et l'intuition. Cette approche factuelle pour la prise de décision indiquée dans la norme ISO 9001, est une partie importante du système de management. Elle est importante pour évaluer l'entreprise et son évolution (taux de substitution et objectif, répartition du chiffre d'affaire par catégorie de produits, référencement de nouvelles gammes de produits) ainsi que l'environnement extérieur (marchés, concurrence, politique). Pour permettre une bonne communication et une implication du personnel, il est important que certaines de ces données leur soit communiquées et qu'ils puissent être informés, voire prendre part aux prises de décisions. Prenons par exemple les données de la CNAM sur les taux de substitution et les objectifs nationaux fixés. Cette information permet à l'entreprise de connaitre ses pratiques de substitution ainsi que les nouvelles molécules inscrites au répertoire des génériques qui ne sont pas encore substituées pour prendre la décision de les référencer en stock ou pas. Au sein de l'officine, peu d'informations sont transmises aux membres de l'équipe, à part le taux de substitution et les objectifs fixés par la CNAM (Caisse Nationale d'Assurance maladie). Les membres de l'équipe ne participent pas à la prise de décisions au sein de l'entreprise, que ce soit des décisions de politique d'entreprise ou de gestion quotidienne (référencement de nouveau produits, aménagements de l'espace de vente et des linéaires) Principe 8 - Les relations mutuellement bénéfiques avec les fournisseurs Un organisme et ses fournisseurs sont interdépendants et des relations mutuellement bénéfiques augmentent leurs capacités à créer une valeur ajoutée. [43] L'entreprise doit établir avec ses partenaires des relations qui équilibrent les gains à court terme et des considérations à long terme. Il est indispensable d'identifier et choisir les fournisseurs clés et de communiquer sur ces partenaires auprès des membres de l'équipe mais éventuellement auprès de la clientèle. Avec ces fournisseurs l'entreprise peut mettre en place des activités communes de développement et d'amélioration. Que ce soit des avantages promotionnels sur des produits ou des services offerts comme des formations, de l'aide au merchandising ou à la mise en place de programme de dépistage (diabète par exemple). Les rapports entre l'entreprise et ses fournisseurs se doivent de suivre les mêmes objectifs, c'est le cas avec notre groupement ainsi que notre fournisseur principal, le grossisterépartiteur CERP Rhin-Rhône-Méditerranée. C'est avec ces partenaires que l'entreprise met en place sa politique de vente (choix des produits, négociation commerciale, mais aussi formation du personnel). La CERP Rhin-Rhône-Méditerranée nous a précédés dans 103 la démarche de certification ISO 9001 : 2000 en l'obtenant en 2004. Cette certification a fait suite à l application des « Bonnes Pratiques de Distribution » (BPD- 30 juin 2000), qui leur ont été opposables dès 2001. Les grossistes ont suivi les laboratoires pharmaceutiques dans leur volonté d'amélioration de la qualité des produits et services mais également dans un objectif de satisfaction client. Au fur et à mesure les officines suivent le pas et l'ensemble de la chaine sera bientôt en assurance qualité. Dans cette chaine chaque maillon est dépendant du maillon précédent, le fournisseur, mais aussi du maillon suivant, le client. Chacun se préoccupe donc des relations avec l'autre, pour finalement obtenir pour le client final le degré de qualité optimal qui lui apportera satisfaction. La mise en place d'une telle démarche au sein de l'officine En conclusion, une pharmacie d'officine doit entrer dans une démarche qualité, et quelque soit sa taille, elle peut le faire en se basant sur la norme ISO 9001 : 2008 comme nous venons de le voir. La norme ISO 9001 : 2008, semble répondre aux exigences actuelles des officines et peut intéresser les grosses structures mais également les petites entreprises. Elle fournit un ensemble d'exigences normalisées pour un système de management de la qualité, indépendamment du domaine d'activité et de la taille de l'organisme utilisateur. Cette norme ISO 9001 : 2008 qui vise à la base la satisfaction client peut devenir tout simplement une norme qui vise la satisfaction générale de l'entreprise : 104 - Satisfaction du client qui obtient le produit et les services attendus selon ses critères de satisfaction - Satisfaction des employés qui travaillent dans des conditions optimales et qui produisent un service qui leur est valorisé - Satisfaction du chef d'entreprise qui travaille avec des collaborateurs impliqués, soucieux de la qualité du travail qu'ils fournissent, ce qui lui permet d'augmenter la satisfaction de ses clients, préambule à l'augmentation de la clientèle et du chiffre d'affaire. Pour aboutir à une éventuelle certification ISO 9001, il est nécessaire de se conformer aux exigences de cette norme ce qui implique qu'il subsiste des difficultés de mise en place. Malgré la rédaction du système documentaire (manuel qualité, cartographie, processus, procédures et documents) il reste des lacunes importantes à notre système de management de la qualité : la définition et la gestion d'indicateurs de performances, d'indicateurs de satisfaction ; le recueil et la gestion des non-conformités, la réalisation et la mise en place des audits internes et des méthodes d'autoévaluations Ces exigences de la norme demandent de nouveau du temps quotidiennement et des connaissances fiques dans le domaine de l'assurance qualité que les pharmaciens n'ont pas forcément. Dans les mois futurs la Pharmacie de Saint Laurent du Pont doit recevoir un organisme certificateur pour une certification à blanc, une ultime rencontre qui nous permettra de remettre en cause ce qui a été fait et ce qu'il reste à faire pour une réelle certification. Quels sont les points importants qui vont permettre la mise en place d'une telle démarche, quelle conclusion nous amène ce travail réalisé dans une pharmacie Laurentinoise? 105 - La qualité c'est un travail d'équipe Une politique d'assurance qualité est le fruit d'un important travail de l'ensemble des membres de l'équipe officinale. Un titulaire ne peut pas imaginer lancer son entreprise dans une démarche qualité sans l'appui et l'engagement de toute son équipe. Pour cela il faut expliquer le concept, les objectifs, l'avenir d'une telle démarche. Chacun doit s'investir dans cette démarche encadrée par le pharmacien titulaire qui est le chef d'orchestre et qui doit entretenir la motivation et l'engagement de l'équipe par des objectifs globaux et personnels d'amélioration. Une politique qualité c'est avant tout une finalité : la satisfaction client. Pour chercher à augmenter cette satisfaction client il faut l'évaluer pour connaitre les points à améliorer, diagnostiquer les problèmes pour chercher des pistes d'amélioration. Il est donc important de connaitre l'avis et le ressenti des clients de l'entreprise pour pouvoir travailler sur des évolutions significative du point de vue du client . - La qualité c'est une vision globale Une entreprise est une entité globale qui fonctionne bien lorsque tous les systèmes fonctionnent bien. La qualité doit s'appliquer à l'ensemble de ses activités : Les processus de réalisation mais aussi les moyens et le management. Ces activités peuvent être schématisées comme on a pu le voir précédemment dans des processus interdépendant les uns des autres. Ces schémas permettent de ne pas alourdir le système documentaire d'innombrables procédures ou modes opératoires qui ne sont pas connus de l'ensemble des salariés. - La qualité demande du temps Mettre en place une démarche qualité, réunir les membres de l'équipe pour un travail en commun, rédiger des procédures, appliquer ces procédures au quotidien demande du 106 temps. Il s'agit d'un temps indispensable mais pas toujours disponible. Il est nécessaire d'établir des priorités sur les futurs évènements sans se focaliser sur une finalité facultative de certification. Il est nécessaire de progresser à son rythme sans se précipiter dans de nombreuses rédactions EXEMPLE DE LA DISPENSATION SUR PRESCRIPTION. L'évolution des pratiques officinales, l'industrialisation du médicament ont entrainé la transformation du « pharmacien qui fabrique et dispense le médicament » en « gestionnaire des thérapeutiques médicamenteuses ». Les exigences croissantes des consommateurs et des organismes sociaux en termes de qualité et de sécurité, les modifications du réseau de distribution, l'accélération de la dégradation des résultats des entreprises officinales face à l'apparition de modèles économiques nouveaux obligent les acteurs de la pharmacie d'aujourd'hui à d'importantes évolutions. Le développement d'une démarche d'assurance qualité apparaît depuis quelques années comme une nécessité pour prouver aux pouvoirs publics, aux organismes sociaux et aux consommateurs que le réseau officinal reste le mieux adapté à ces exigences. Cette démarche est largement recommandée par les différentes instances officielles (Ordre des Pharmaciens, syndicats de pharmaciens, Haut Conseil pour l'avenir de l'Assurance Maladie). La reconnaissance de cette implication dans cette démarche est la certification de l'entreprise par un organisme officiel. Le présent travail réalisé à la Pharmacie de Saint Laurent du Pont en Isère a pour objectif le développement de la démarche d'assurance qualité afin de l'emmener à une certification ISO 9001. La partie du travail relaté dans ce manuscrit s'oriente sur la réalisation d'un processus de dispensation sur prescription au sein de l'entreprise. En termes de méthodologie, nous avons, à travers un processus de type Delphi, fait participer l'ensemble de l'équipe officinale. Les avis et observations de chacun sont 109 recueillis de manière anonyme et mis en commun de manière itérative jusqu'à obtenir un consensus. Grace aux résultats finalement obtenus, nous rédigeons le processus de dispensation sur prescription ainsi que les procédures qui encadrent ce processus. Par l' ntermédiaire de cette démarche l'ensemble des activités de l'entreprise officinale peuvent être formalisées selon la norme ISO 9001, afin de révéler au patientconsommateur notre volonté de qualité dans notre exercice quotidien. Mais la « qualité » n'est pas forcément la norme. Il peut exister une importante politique d'Assurance Qualité dans une entreprise sans vouloir intégrer la norme ISO 9001. La politique d'assurance qualité d'une officine ne poursuit pas l'unique but d'une certification (qui demande une importante mobilisation de ressources humaines et financières). Elle permet surtout l'adaptation de nos méthodes de travail aux besoins et exigences de nos clients. Au terme de ce travail, il nous apparaît que la mise en oeuvre d'une démarche Qualité, s'inscrit dans notre réflexion sur la valeur ajoutée du pharmacien d'officine dans la chaîne du médicament. Grenoble LE DO LE PRESIDENT DU JURY Docteur Benoit ALLENET 110 ANNEXES 111 ANNEXE N°1 : PREMI ERE ETAPE DELPHI 112 dispensation Delphi.étape1 identification du patient concerné par l'ordonnance : - demande de renseignement s sup plémentaires ( age , poids pour les enfants ) - Recherche des caractéristiques physiopathologiques du patient (femme enceinte, femme allai tant, personne en fin de vie) BESOINS items clairement dans les étapes suivantes. Elles ont été classées par ordre chronologique et par étapes importantes pour pouvoir apparaître Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc vérification des posologies (en utilisant le Vidal informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise Recherche des éventuelles interactions médicamenteuses identification du prescripteur : - Généraliste, spécialiste, hôpital - Médecin connus (médecins du canton ) lecture complète et attentive de la prescription identification d'une première délivrance ou d'un renouvellement demander au patient s'il y a un changement dans son traitement chronique (historique thérapeutique) identification des objectifs thérapeutiques visés par la prescription Interrogation sur la cohérence du traitement prescrit Interrogation sur le statut réglementaire des différents produits (liste, anxiolytiques, hypnotiques, stupéfiants, PIH, MDS, médicaments à prescriptions restreintes) ouvrir la page historique pour visualiser et comparer avec les anciennes prescriptions Interrogation sur l'existence de produits à surveillance particulière (marge thérapeutique étroite, bilan bio initial, suivi biologique important) Se renseigner sur le suivi biologique du traitement (INR, Kaliémie, HbA1c, suivi glycémique, ) Demander au patient ses analyses biologiques En cas de découverte d'une anomalie: - interrogation d'un pharmacien - appel au prescripteur pour confirmation si nécessaire dispensation Delphi.étape1 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique - question au sur son accord de substitution classement mental des différents produits de la prescription par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement vérification des dosages des produits par rapport aux dosages prescrit (risques de confusions) dispensation Delphi.étape1 accueil du client à son entrée dans l'officine accueil personnalisé du patient si possible annoncer au patient ce que l'on fait (ex : je vais chercher vos médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet dispensation Delphi.étape1 dispensation delphi2.phase2 BESOINS items 1 prescription. commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc vérification des posologies (en utilisant le Vidal informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise Recherche des éventuelles interactions médicamenteuses - dispensation delphi2.phase2 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique - question au patient sur son accord de substitution classement mental des différents produits de la prescription par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement vérification des dosages des produits par rapport aux dosages prescrit (risques de confusions) dispensation delphi2.phase2 accueil du client à son entrée dans l'officine accueil personnalisé du patient si possible annoncer au patient ce que l'on fait (ex : je vais chercher vos médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquette s posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet Vandenheede marc Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie dispensation delphi2.phase2 dispensation delphi2.phase2 - resultats1 - PH2 BESOINS items X X X X X X X X X 1 4 prescription. De 1 : la plus grande importance à 4 la moins grande importance. commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc X vérification des posologies (en utilisant le Vidal informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise X Recherche des éventuelles interactions médicamenteuses X - X X X X X X dispensation delphi2.phase2 - resultats1 - PH2 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE X X X X X X X interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique X - question au patient sur son accord de substitution X classement mental des différents produits de la prescription par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement X vérification des dosages des produits par rapport aux dosages X prescrit (risques de confusions) X X dispensation delphi2.phase2 - resultats1 - PH2 accueil du client à son entrée dans l'officine accueil personnalisé du patient si possible annoncer au patient ce que l'on fait (ex : je vais chercher vos médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d' utilisation des produits : - concern ant tous les produits de prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet X X X X X SI BESOIN SI BESOIN SI BESOIN SI BESOIN Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie? dispensation delphi2.phase2 - resultats1 - PH2 dispensation delphi2.phase2 - resultats1 - PH3 BESOINS items X X X X X X X X prescription. commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc X X X X X X vérification des posologies (en utilisant le Vidal X informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise X Recherche des éventuelles interactions médicamenteuses X - X X dispensation delphi2.phase2 - resultats1 - PH3 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE X facturation des produits : - dans l'ordre de la prescription X - en respectant les zones ALD et non-ALD X impression de la facture (ou non si renouvellement sécurisé) X impression de l'ordonnancier sur original de la prescription X En cas de produit manquant, réalisation d'un bon de promis X Mise en place de la traçabilité réglementaire de certains X produits (médicaments dérivés du sang, médicaments à prescriptions restreintes, stupéfiants) vérifier nombre de boites délivrées par rapport au nombre de X boites facturées ENREGISTREMENTS - FACTURATION interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique X - question au patient sur son accord de substitution X classement mental des différents produits de la prescription X par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement X vérification des dosages des produits par rapport aux dosages X prescrit (risques de confusions) dispensation delphi2.phase2 - resultats1 - PH3 X X X X X X X X X accueil du client à son entrée dans l'officine X accueil personnalisé du patient si possible X annoncer au patient ce que l'on fait (ex : je vais chercher vos X médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose X Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet Vandenheede marc Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie? dispensation delphi2.phase2 - resultats1 - PH3 dispensation delphi2.phase2 - resultats1 - Prep1 BESOINS items commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc X X X vérification des posologies (en utilisant le Vidal X informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise X Recherche des éventuelles interactions médicamenteuses X - X X Non concerné -> Pharmacien dispensation delphi2.phase2 - resultats1 - Prep1 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE X X X X X X interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique X - question au patient sur son accord de substitution X classement mental des différents produits de la prescription X par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement X vérification des dosages des produits par rapport aux dosages X prescrit (risques de confusions) X X dispensation delphi2.phase2 - resultats1 - Prep1 dispensation delphi2.phase2 - resultats1 - Prep1 accueil du client à son entrée dans l'officine X accueil personnalisé du patient si possible X annoncer au patient ce que l'on fait (ex : je vais chercher vos X médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose X Communication: - Si le porteur de l'ordonnance n'est pas le consommateur X Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie? final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement X pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription X - concernant les produits à usage particulier s (ATB, X Asthme) description au patient de la posologie X Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement X - si posologie prescrite est erronée X évaluer le niveau de compréhension du patient X re-vérification des produits délivrés par rapport à la X prescription au moment de mettre dans sachet Vandenheede marc dispensation delphi2.phase2 - resultats1 - Prep2 BESOINS items X X X X X prescription. commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc vérification des posologies (en utilisant le Vidal informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise Recherche des éventuelles interactions médicamenteuses - X X X X X X X X X X X X X X X X dispensation delphi2.phase2 - resultats1 - Prep2 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE X facturation des produits : - dans l'ordre de la prescription X - en respectant les zones ALD et non-ALD X impression de la facture (ou non si renouvellement sécurisé) X impression de l'ordonnancier sur original de la prescription X En cas de produit manquant, réalisation d' un bon de promis X Mise en place de la traçabilité réglementaire de certains X produits ( médicaments dérivés du sang , médicaments à prescription s restreintes, stupé fiant s) vérifier nombre de boites délivrées par rapport au nombre de X boites facturées interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique X - question au patient sur son accord de substitution X classement mental des différents produits de la prescription X par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement X vérification des dosages des produits par rapport aux dosages X prescrit (risques de confusions) dispens ation delphi2.phase2 - resultats1 - Prep2 X X X X X X X accueil du client à son entrée dans 'officine X accueil personnalisé du patient si possible X annoncer au patient ce que l'on fait (ex : je vais chercher vos X médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose X Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement (Observance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet X X X Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie? dispensation delphi2.phase2 - resultats1 - Prep2 dispensation delphi2.phase2 - resultats1 - Prep3 BESOINS items X X X X X X X Seulement si droits perimés prescription. commentaires résultat de leur avis. Ils ont répondu en fonction de l'importance estimée des différents items lors d'une dispensation de produits sur Après avoir répertorié les différents items obtenus lors de l'interrogation des salariés autorisés à la dispensation, ce tableau est le Vandenheede marc X vérification des posologies (en utilisant le Vidal X informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise X Recherche des éventuelles interactions médicamenteuses X - X X X X X Selon les cas dispensation delphi2.phase2 - resultats1 - Prep3 Énumération des différents médicaments en indiquant de quoi il s'agit - Si le porteur de l'ordonnance est le consommateur final de la prescription DELIVRANCE X X X X X X X X interrogation sur une éventuelle substitution (appartenance d'un ou plusieurs produits au répertoire des génériques) : - utilisation de l'historique X - question au patient sur son accord de substitution X classement mental des différents produits de la prescription par ordre alphabétique pour faciliter leur recherche en tiroir recherche des spécialités dans l'ordre de la prescription calcul des quantités nécessaires et suffisantes au traitement X vérification des dosages des produits par rapport aux dosages X prescrit (risques de confusions) X Surtout si c'est un nouveau traitement dispensation delphi2.phase2 - resultats1 - Prep3 X X X X X accueil du client à son entrée dans l'officine X accueil personnalisé du patient si possible X annoncer au patient ce que l'on fait (ex : je vais chercher vos médicaments, je recherche des informations sur ce produit ) demander au patient s'il souhaite autre chose X Communication: - Si le porteur de l'ordonnance n'est pas le consommateur final (problème du secret professionnel) Interroger le patient sur sa façon de prendre son traitement pour vérifier son niveau d'adhésion au traitement ( servance) Description du mode d'utilisation des produits : - concernant tous les produits de la prescription - concernant les produits à usage particuliers (ATB, Asthme) description au patient de la posologie Réaliser des étiquettes de posologies : - tout le temps - si patient âgés - si nourrisson ou enfants - si nouveau traitement - si posologie prescrite est erronée évaluer le niveau de compréhension du patient re-vérification des produits délivrés par rapport à la prescription au moment de mettre dans sachet X X X X X X X Si utilisation particulière Dans ce cas édition d'un plan de prise ou d'étiquettes de posologie? dispensation delphi2.phase2 - resultats1 - Prep3 dispensation delphi2.phase2 - resultats2 - PH2 BESOINS items X par leurs homologues. Ici apparaissent les résultats de l'étape suivante, les experts ont pu modifier leurs réponses en fonction des réponses les plus citées Vandenheede marc X X X X X X X X X X X X X X X vérification des posologies (en utilisant le Vidal X informatique si nécessaire) Vérification des rythmes d'administration et du plan de prise X Recherche des éventuelles interactions médicamenteuses X - X X Contrôle supplémentaire TOUJOURS. Interroger un supérieur ou un confrère En faire la demande afin de provoquer un dialogue.	48,504
https://openalex.org/W2621625341	OpenAlex	Open Science	CC-By	2,017	Atividade física e tempo sentado combinados e sua contribuição no índice de massa corporal em adultos	Priscila Bezerra Gonçalves	English	Spoken	5,335	10,373	Atividade física e tempo sentado combinados e sua contribuição no índice de massa corporal em adultos Priscila Bezerra Gonçalves1,2 Adalberto Aparecido dos Santos Lopes1,2 Alexandre Augusto de Paula Silva1,2 Jeruza Sech Buck Silva1,2 Alice Tatiane da Silva1,2 Leonardo Augusto Becker1,2 Marilson Kienteka1,2 Rodrigo Siqueira Reis1,2,3 Priscila Bezerra Gonçalves1,2 Adalberto Aparecido dos Santos Lopes1,2 Alexandre Augusto de Paula Silva1,2 Jeruza Sech Buck Silva1,2 Alice Tatiane da Silva1,2 Leonardo Augusto Becker1,2 Marilson Kienteka1,2 Rodrigo Siqueira Reis1,2,3 Abstract – The aim of this study was to analyze the contribution of physical activity (PA) and sedentary time (ST) to Body Mass Index (BMI) in adults. The study was conducted in 2009, in Curitiba, Parana, Brazil with adults aged 18-65 years (1,411). A multidimen- sional questionnaire was used in which BMI, weekly minutes of PA and ST were obtained from self-reports. PA and ST were combined to determine the following independent variables: a) “active and not sedentary”; b) “active and sedentary”; c) “not active and not sedentary” and d) “not active and sedentary”. Multinomial logistic regressions were used to test the associations between PA, ST and BMI. The adjusted regression model showed that PA, regardless of ST, lowered the likelihood of participants being classified as obese by 34% and 55%, respectively. In addition, PA at recommended levels (≥150 minutes/ week), while not being sedentary, lowered by 85% the likelihood of being classified in the underweight category. Finally, ≥10 minutes/week of PA, while reporting higher ST levels, reduced the likelihood of being classified as overweight by 37%. PA is associated with BMI regardless of TS. In addition, PA at recommended levels is inversely related to obesity and overweight. Key words: Brazil; Motor activity; Nutritional status; Sedentary lifestyle. Resumo – O objetivo do estudo foi analisar a contribuição da atividade física (AF) e do tempo sedentário (TS) no Índice de Massa Corporal (IMC). O estudo foi conduzido em 2009, em Curitiba, Paraná, Brasil com adultos de 18 à 65 anos (n=1.411). Foi aplicado um questionário multidimensional em que o IMC, AF e TS foram obtidos de forma autorreportada. AF e TS foram combinados para determinar as variáveis independentes: a) “ativo e não sedentário”; b) “ativo e sedentário”; c) “não ativo e não sedentário” e d) “não ativo e sedentário”. A regressão logística multinominal foi empregada para testar as associações entre AF, TS e IMC. As análises ajustadas demonstraram que AF, independente do TS, reduz entre 34% e 55% as chances dos participantes serem classificados como obesos. 1 Pontifical Catholic University of Parana. Research Group on Physical Activity and Quality of Life. Curitiba, PR. Brazil. 2 Federal University of Parana. Gra- duate Program in Physical Education. Curitiba, PR. Brazil. original article original article Palavras-chave: Atividade motora; Brasil; Estado nutricional; Estilo de vida sedentário. Rev Bras Cineantropom Hum Rev Bras Cineantropom Hum INTRODUCTION High Body Mass Index (BMI) values are an important risk factor for chronic noncommunicable diseases (NCDs), such as cardiovascular diseases, renal diseases, diabetes, some cancers and musculoskeletal disorders 1. Data from more than 19 million people in 186 countries have shown that the mean BMI has increased considerably in recent decades1. In Brazil, in 2014, the prevalence of adults in Brazilian capitals with overweight and obesity was 52.5% and 17.9%, respectively2. Thus, the identification of factors that may attenuate or reverse this situation has been widely discussed in literature3. In this sense, the regular practice of physical activity (PA) and the re- duction of sedentary time (ST) have been highlighted throughout the world as they are factors of prevention for the emergence of NCDs4. Nevertheless, 23% of the world population5 and 15.4% of the Brazilian population2 is considered physically inactive. Regarding sedentary behavior, 25.3% of the adult population of Brazilian capitals has a habit of watching television for more than three hours daily2. In addition, women, older adults and people of low socioeconomic status (SES) are more likely to spend longer times in sedentary behavior6.h There is evidence, especially in high-income countries, that the increase in PA levels and the reduction of ST are associated, independently and together, with lower adiposity7 and also with favorable lipid profile (HDL cholesterol)8. Data from a multicenter study in 10 countries have shown that higher PA levels were associated with reduction in the probability of overweight and obesity in adults, although no significant results were found for ST9. In addition, a recent meta-analysis has found that practicing at least 60 minutes of moderate daily PA can eliminate the increased risk of death for a variety of NCDs, such as dyslipidemia, diabetes, hypertension, and cardiovascular disease caused by high ST10. In a cohort study conducted in Switzerland with 3,042 adults between 1991 and 2011, vigorous PA was inversely associated with obesity in both cross-sectional and longitu- dinal data. Participants who remained or became inactive were associated with weight gain and increased body fat, but ST was not associated with increased BMI11. In another study conducted in Brazil, active commuting and BMI were inversely associated12. Despite the consistent relationship between influence of PA on BMI and the benefits of regular practice of PA, to date, there is no evidence of the combined association of PA and ST with population BMI in samples of the Brazilian population. Rev Bras Cineantropom Desempenho Hum 2017, 19(2):174-184 Atividade física e tempo sentado combinados e sua contribuição no índice de massa corporal em adultos Além disto, praticar ≥150 minutos/semana de AF e não ser sedentário reduziu em 85% as chances dos participantes serem classificados com baixo peso. Por fim, praticar ≥10 minutos/semana de AF, mesmo apresentando TS elevado, reduziu em 37% a chance de classificação em sobrepeso. A prática de AF está associada com IMC independente do TS. Além disto, AF em níveis recomendados tem relação inversa com a obesidade e sobrepeso. 2 Federal University of Parana. Gra- duate Program in Physical Education. Curitiba, PR. Brazil. 3 Washington University in St. Louis. Prevention Research Center. Brown School. St. Louis, MO. USA. 3 Washington University in St. Louis. Prevention Research Center. Brown School. St. Louis, MO. USA. Received: 11 January 2017 Accepted: 10 March 2017 Received: 11 January 2017 Accepted: 10 March 2017 Licença Creative Commom BY BY BY vras-chave: Atividade motora; Brasil; Estado nutricional; Estilo de vida sedentário. INTRODUCTION INTRODUCTION Available studies have analyzed these behav- iors and factors in isolation13. Understanding this relationship will serve as a subsidy for public health decision makers, especially because in the last 10 years, in the Brazilian population, there was a 10% increase in the prevalence of overweight and obesity, 16% of physical inactivity and 23% in daily time watching television (≥3 hours / day). Thus, the aim of this study was to analyze the contribution of PA and ST, analyzed together, on BMI levels in an adult sample. 175 Gonçalves et al. Gonçalves et al. Physical activity, sitting time and BMI Place of studyh This study was carried out in the city of Curitiba, capital of the State of Paraná, which is located in the southern region of Brazil. At the time of data collection, the city had 1,897,997 inhabitants (≈1,130,139 adults) in an area of 435,036 Km2. The city has high Municipal Human Develop- ment Index (0.823), when compared to other Brazilian municipalities, and with an extensive green area (65m2 per inhabitant), which includes 21 parks and 454 squares14. Selection of parks and squares For the present study, eight public leisure spaces (four parks and four squares) were selected. Initially, neighborhoods in which there were pub- lic spaces with characteristics of the physical environment were selected (e.g., parks, squares, animation axes) suitable for the practice of PA (75 neighborhoods). Then, with the aim of covering different characteristics of neighborhoods and the population of Curitiba, neighborhoods were stratified according to income and characteristics of the “social and built environment” (AMB) aimed at the practice of PA16. Coordinators of the Municipal Secretariat of Sports Leisure and Youth (SMELJ) and Municipal Health Secretariat (SMS), indicated in which public spaces there were PA interventions or programs. After three consultations, consensus was ob- tained among coordinators and four parks and four squares were selected, being distributed in different income strata and AMB (high income and high AMB, high income and low AMB, low income and high AMB; low income and low AMB)14. Design, study characteristics and ethical aspectsh This is an observational epidemiological study of cross-sectional design, part of a broader project called “Caminhos para o Parque”, which aimed to examine the role of urban parks in PA levels of adults in Curitiba, Paraná, Brazil. The details of the methodology of this study are described in literature14. Selection of households and participants For the selection of households, an area of 500 meters radius was deter- mined from the central point of each public space included in the study. All segments of streets within this area were listed in order to identify places with domiciles. In each street segment, a household was randomly selected, and all residents of the household considered eligible (≥18 years, apparently healthy and residing at the place selected for at least one year) were identified. Among these, a resident was randomly selected. More detailed information on the sampling process is available in literature14. Covariates Covariates were obtained through a questionnaire. Sex was obtained by the observation of interviewers (male / female). The age group was calculated from the date of birth and classified into five categories (18-29 years, 30-39 years, 40-49 years, 50-59 years and ≥60 years). Finally, SES was evaluated with questions that consider the number of household appliances, presence of maid and schooling of the family head, proposed by the Brazilian As- sociation of Research Companies (ABEP). For purposes of analysis, SES was classified into three categories: low (E, D, C2 and C1), intermediate (B2 and B1) and high (A2 and A1). Rev Bras Cineantropom Desempenho Hum 2017, 19(2):174-184 Dependent variable BMI was calculated from self-reported measures of height and body mass17 and participants were classified as “low weight” (≤18.5 kg / m²), “normal weight” (18.6-24.9 kg / m²), “overweight” (25-29.9 kg / m²) and “obese” (≥30 kg / m²), as recommended by the World Health Organization18. Independent variables Total leisure PA was subjectively measured using the long version of the International Physical Activity Questionnaire (IPAQ ), validated for the Brazilian population, using the following equation: [walk + moderate ac- tivity + (vigorous activity * 2)]19. ST was measured by the sum of the time spent seated during weekdays and weekends, except time spent sitting in a passive commuting (bus and / or car)4. Subsequently, the daily mean of the total time spent in ST was calculated. For purposes of analysis, two models were created for the combination of variables: a) active: PA ≥150 minutes / week and sedentary: ST ≥240 minutes / day, according to PA20 and ST recommendations4,10; b) active: PA ≥ 10 minutes / week and sedentary: TS ≥240 minutes / day, consider- ing a less rigid criterion for PA and ST recommendations4,10. Finally, the independent variable was operationalized for each model, with PA and ST combination: a) “active and not sedentary”; b) “active and sedentary”; c) “not active and not sedentary” and d) “not active and sedentary”. The combination of these variables allowed us identifying which behaviors help to obtain a more adequate nutritional profile. Data collection Face-to-face interviews were conducted between April and July 2009, 176 totaling 86 days. Twenty-five female interviewers with experience in data collection were selected and trained by specialists from the Research Group on Physical Activity and Quality of Life. The theoretical course lasted 24 hours and the practical one lasted four hours. The research was approved by the Ethics Research Committee (No. 005/2008) and participants signed the Free and Informed Consent Form. RESULTS Of the total sample, 50 participants were excluded because they did not present complete data of variables used in the present study. Thus, the final sample consisted of 1,411 adults, mostly women (63.1%), aged 50-59 years (22.8%), intermediate SES (49.5%) and BMI classified as normal (49.1%). Overall, 35.2% of participants reported performing ≥150 minutes / week of leisure PA and more than half (54.7%) reported practicing ≥10 minutes / week of leisure PA. In relation to ST, it was observed that in out of 10 par- ticipants, four remained ≥240 minutes / day in this behavior (43.2%). When considering the PA and ST categories, a greater proportion was observed in the non-sedentary and non-active groups for both PA classifications, ≥150 minutes / week (37.8%) and ≥10 minutes / week (26.4 %) (Table 1).h The most frequent measures among men were ST (48.4%, p = 0.003), overweight (37.8%, p = 0.016) and PA level considered active for both clas- sifications, ≥150 minutes / week (41.8%, p <0.001) and ≥10 minutes / week (63.3%, p <0.001). On the other hand, low weight (3.3%, p = 0.016) and obesity (15.5%, p = 0.016) were more frequent among women. Regarding age group, higher frequency of adults classified as sedentary (57.6%, p <0.001), with low weight (3.8%, p <0.001) and normal weight (68.0%, p < 0.001) was found among participants aged 18-29 years. Finally, for SES, higher fre- quency of participants classified as sedentary (54.4%, p <0.001) and active in both classifications, ≥150 minutes / week (55.0%, p <0.001) and ≥10 minutes / week (75.6%, p <0.001) was found in participants with high SES (Table 2). In the multinomial logistic regression analysis, for the combination of BMI, PA performed at ≥150 minutes / week and ST (Figure 1), there was an inverse association between “low weight” and “active / not sedentary” (OR: 0. 15, 95% CI 0.03-0.67), “obese” and “active / sedentary” (OR: 0.45, 95% CI: 0.26-0.77) and “obese” and “active / not sedentary “(OR: 0.52; 95% CI: 0.32-0.48) when adjusted for sex, age and SES. Statistical analysis Absolute and relative frequency distribution was performed to character- ize the sample. Subsequently, the chi-square test (x2) for heterogeneity 177 Gonçalves et al. Gonçalves et al. Physical activity, sitting time and BMI was used to test the association between covariates and the PA and ST models. After performing bivariate analyses, covariates that presented p≤0.20 were included in the adjusted model (sex, age group and SES), as possible confounding factors21. Finally, gross and adjusted multinomial logistic regression was used to test the association between PA and ST combinations with BMI classifications. The forced input method was used in which all previously tested significant covariates (p≤0.20) were inserted into the regression model. Analyses were performed in SPSS 23.0 and the significance level was maintained at 5%. RESULTS Variables Categories n % Sex Male 521 36.9 Female 890 63.1 Age 18-29 316 22.4 30-39 276 19.6 40-49 317 22.5 50-59 322 22.8 ≥60 180 12.8 Socioeconomic status Low 532 37.7 Intermediate 699 49.5 High 180 12.8 Body mass index Low weight (≤18.5) 37 2.6 Normal weight (18.6–24.9) 693 49.1 Overweight (25.0–29.9) 470 33.3 Obese (≥30.0) 211 15.0 Physical activity * Inactive (0-149 min/week) 914 64.8 Active (≥150 min/week) 497 35.2 Physical activity ** Inactive (0-9 min/week) 639 45.3 Active (≥10 min/week) 772 54.7 Sedentary time Not sedentary (0-239 min/week) 801 56.8 Sedentary (≥240 min/day) 610 43.2 Categories sedentary time with physical activity * Not active and sedentary 381 27.0 Not active and not sedentary 533 37.8 Active and sedentary 229 16.2 Active and not sedentary 268 19.0 Categories sedentary time with physical activity ** Not active and sedentary 267 18.9 Not active and not sedentary 372 26.4 Active and sedentary 343 24.3 Active and not sedentary 429 30.4 Min=minutes; Week=week; ≥150 minutes/week; ≥10 minutes/week . Min=minutes; Week=week; ≥150 minutes/week; ≥10 minutes/week *. Table 2. Association between individual characteristics and physical activity, sedentary time and body mass index of adults enrolled in the “Caminhos para o parque” project. Curitiba, Paraná, Brazil. (n = 1411). Table 2. Association between individual characteristics and physical activity, sedentary time and body mass index of adults enrolled in the “Caminhos para o parque” project. Curitiba, Paraná, Brazil. (n = 1411). RESULTS For the combination of BMI, PA performed at ≥10 minutes / week and ST (Figure 2), there was an inverse association between “low weight” and “active / not sedentary” (OR: 0.35, 95% CI: 0.13-0.93), “overweight” and “active / sedentary” (OR: 0.63, 95% CI: 0.43-0.92), “obese” and “active / sedentary” (OR: 0.56; 95% CI: 0.35-0.92), “obese” and “active / sedentary” (OR: 0.52; 95% CI: 0.33- 0.82) when adjusted for sex, age and SES. 178 Table 1. Sociodemographic characteristics, physical activity and sedentary time in adults enrolled in the “Caminhos para o Parque” project. Curitiba, Paraná, Brazil (n = 1.411). in the Caminhos para o Parque project. Curitiba, Paraná, Brazil (n = 1.411). Variables Categories n % Sex Male 521 36.9 Female 890 63.1 Age 18-29 316 22.4 30-39 276 19.6 40-49 317 22.5 50-59 322 22.8 ≥60 180 12.8 Socioeconomic status Low 532 37.7 Intermediate 699 49.5 High 180 12.8 Body mass index Low weight (≤18.5) 37 2.6 Normal weight (18.6–24.9) 693 49.1 Overweight (25.0–29.9) 470 33.3 Obese (≥30.0) 211 15.0 Physical activity Inactive (0-149 min/week) 914 64.8 Active (≥150 min/week) 497 35.2 Physical activity ** Inactive (0-9 min/week) 639 45.3 Active (≥10 min/week) 772 54.7 Sedentary time Not sedentary (0-239 min/week) 801 56.8 Sedentary (≥240 min/day) 610 43.2 Categories sedentary time with physical activity * Not active and sedentary 381 27.0 Not active and not sedentary 533 37.8 Active and sedentary 229 16.2 Active and not sedentary 268 19.0 Categories sedentary time with physical activity ** Not active and sedentary 267 18.9 Not active and not sedentary 372 26.4 Active and sedentary 343 24.3 Active and not sedentary 429 30.4 Min=minutes; Week=week; ≥150 minutes/week; ≥10 minutes/week . Table 2. Association between individual characteristics and physical activity, sedentary time and body mass index of adults enrolled in the “Caminhos para o parque” project. Curitiba, Paraná, Brazil. (n = 1411). Rev Bras Cineantropom Desempenho Hum 2017, 19(2):174-184 RESULTS Variables/ Categories Sedentary time Physical activity Body mass index Not sedentary Sedentary Not active Active Not active Active Low weight Normal weight Over- weight Obese n(%) n(%) p n(%) n(%) p n(%) n(%) p n(%) n(%) n(%) n(%) p Sex Male 262(51.6) 252(48.4) 0.003a 303(58.2) 218(41.8) <0.001a 191(36.7) 330(63.3) <0.001a 8(1.5) 243(46.6) 197(37.8) 73(14.0) 0.016a Female 532(59.8) 358 (40.2) 611(68.7) 279(31.3) 448(50.3) 442(49.7) 29(3.3) 450(50.6) 273(30.7) 138(15.5) Age 18-29 134(42.4) 182(57.6) <0.001a 197(62.3) 119(37.7) 0.409a 145(45.9) 171(54.1) 0.220a 12(3.8) 215(68.0) 69(21.8) 20(6.3) <0.001a 30-39 161(58.3) 115(41.7) 184(66.7) 92(33.3) 112(46.6) 164(59.4) 3(1.1) 144(52.2) 86(31.2) 43(15.6) 40-49 182(57.4) 135(42.6) 207(68.5) 100(31.5) 159(50.2) 158(49.8) 10(3.2) 130(41.0) 129(40.7) 48(15.1) 50-59 203(63.0) 119(37.0) 201(62.4) 121(37.6) 144(44.7) 178(55.3) 10(3.1) 130(40.4) 114(35.4) 68(21.1) ≥60 121(67.2) 59(32.8) 115(63.9) 65(36.1) 79(43.9) 101(56.1) 2(1.1) 74(41.1) 72(40.0) 32(17.8) Socioeconomic status Low 353(66.4) 179(33.6) <0.001a 396(74.4) 136(25.6) <0.001a 298(56.0) 234(44.0) <0.001a 16 (3.0) 238(44.7) 188(35.3) 90(16.9) 0.167a Intermediate 366(52.4) 333(47.6) 437(62.5) 262(37.5) 297(42.5) 402(57.5) 15 (2.1) 358(51.2) 225(32.2) 101(14.4) High 82(45.6) 98(54.4) 81(45.0) 99(55.0) 44(24.4) 136(75.6) 6 (3.3) 97(53.9) 57(31.7) 20(11.1) * ≥240 minutes of sedentary time per day; ≥150 minutes of physical activity per week (leisure); * ≥10 minutes of physical activity per week (leisure); a Chi-square test; p≤0.05 (significance) Rev Bras Cineantropom Desempenho Hum 2017, 19(2):174-184 179 Physical activity, sitting time and BMI Gonçalves et al. Figure 1. Box-plot for multinomial logistic regression analysis, crude and adjusted, between BMI and combination of physical activity (guidelines) and sedentary time (n=1,411). BMI: Body Mass Index; Active: ≥150 minutes/week; Sedentary: ≥240 minutes/day; Adjusted by sex, age group and socioeconomic status; p≤0.05. Physical activity, sitting time and BMI (guidelines) and sedentary time (n=1,411). BMI: Body Mass Index; Active: ≥150 minutes/week; Sedentary: ≥240 minutes/day; Adjusted by sex, age group and socioeconomic status; p≤0.05. Figure 2. Box-plot for multinomial logistic regression, crude and adjusted, analysis between BMI and combination of physical activity (>10minutes/week) and sedentary time (n=1,411). BMI: Body Mass Index; Active: ≥10 minutes/week; Sedentary: ≥240 minutes/day; Adjusted by sex, age group and socioeconomic status; p≤0.05. Figure 2. Box-plot for multinomial logistic regression, crude and adjusted, analysis between BMI and combination of physical activity (>10minutes/week) and sedentary time (n=1,411). BMI: Body Mass Index; Active: ≥10 minutes/week; Sedentary: ≥240 minutes/day; Adjusted by sex, age group and socioeconomic status; p≤0.05. DISCUSSION The aim of this study was to test the contribution of PA and ST, analyzed together, in the BMI of Brazilian adults. The main results showed that the likelihood of people being classified as obese and underweight when compared to normal weight was lower in groups that practiced PA, re- gardless of the weekly volume. In addition, practicing minimum PA (≥10 minutes / week) decreased the participants’ likelihood of being classified as overweight. Regarding individual characteristics, only one-third of the interviewees complied with recommendations of practice of leisure PA and approximately half of the sample was classified as overweight or obese. In addition, achieving recommendations of practicing ≥150 minutes of PA per week20 was associated with males (42%) and with high SES (55%). Similar results were identified in adults living in the Brazilian capitals, where 37.6% of this population performed leisure PA, being more prevalent among men (45.6%) aged 18-24 years (51.4%)2. In relation to ST, there was an inverse association with age group, that is, the younger (18-29 years), the higher the ST, corroborating results of a study carried out in 20 countries, 180 in which adults aged 40-65 years were less likely of showing this behavior when compared to the age group of 18-39 years22.i In this study, participants classified as low weight, those who prac- ticed some weekly PA and did not remain four hours a day or more in ST, were less likely of being low weight when compared to not active or active groups with ST greater than four hours daily. These findings reinforce the importance of PA, even in small volumes, since low weight is a health con- dition responsible for an 80% increase in the risk of death from any cause compared to normal weight23. In addition, the causes of low weight may be related to excessive alcohol consumption, smoking and inadequate diet24, but these relationships still remain inconclusive. However, it is known that the practice of PA can influence changes in lifestyle and healthy habits, reducing the risk factors of low weight and increasing the likelihood of participants achieving an ideal weight3,5. Participants who had ≥ 10 minutes of leisure PA, even with high ST, were less likely of being overweight compared to those classified as not active and sedentary. Evidence suggests that excess ST is an important risk factor for chronic diseases and premature mortality25. Rev Bras Cineantropom Desempenho Hum 2017, 19(2):174-184 CONCLUSION In this study, PA was shown to be a protective factor against overweight and obesity, regardless of ST volume. This evidence strengthens the role of PA in controlling obesity, particularly for people who spend long time performing seated activities, such as in some jobs and during motor trans- portation. Obesity control actions should prioritize increased opportuni- ties for practicing PA in work environments, as well as increasing access to healthier commuting, such as walking and cycling. In addition, future studies should be conducted on samples from other regions and cities, as well as objective measures should be used to evaluate the PA and ST in order to confirm the findings of this study. Physical activity, sitting time and BMI even if people have high ST. For a better interpretation of results, some limitations should be con- sidered. The cross-sectional design does not allow establishing a causal relationship among variables. Self-reported measures tend to overestimate PA and ST and may contribute to an increase in non-differential error. In the analyses, only leisure PA was considered, not allowing extrapolation of the findings to other domains of PA. Sample selection was conducted in regions intentionally selected based on the social and physical characteristics of the dwelling region of participants. However, the sample distribution was balanced among neighborhoods, reducing the chances of clustering effect. In addition, the main confounding variables were included in the adjusted analyses and the effects were analyzed along different exposure and outcome gradients, assuring greater confidence in the internal validity of the findings. DISCUSSION Thus, PA can also act as a protective factor, mitigating the detrimental effects of ST. These findings corroborate more recent literature, indicating that performing at least one hour of PA per day may reduce the risk of death associated with high sitting time10.i More significant results were found for the group of obese participants. Performing some weekly PA regardless of classification used (≥ 10 minutes / week or ≥150 minutes / week), represented approximately 50% less chance of participants being classified as obese, regardless of ST, reinforcing the findings of this study on the importance of the practice of PA and reduction of ST. Literature shows that physical inactivity was responsible for 6% of the causes of death worldwide3 and 3.8% were attributed to sedentary behavior 26. In addition, the decrease in physical inactivity and ST can increase life expectancy, on average, 0.68 and 0.23 years, respectively 3,26. These results indicate that changes in these behaviors, especially in the practice of PA, seem to be the starting point for maintaining healthy body weight.h The results of this study highlight the importance of multifactorial actions to promote healthy habits, such as the inclusion of PA in daily life3,27, healthy eating27 and body weight control1. In addition, public poli- cies should consider actions in community settings that may benefit, in particular, less active groups with higher prevalence of ST28. For example, propose changes in the planning of cities that favor the practice of PA through active commuting 29. In general, it seems that the inclusion of PA in people’s daily lives, even in reduced volume, can improve the chances of adjusting BMI to optimal health levels, thereby improving quality of life and self-esteem, and reducing costs of the public health system30. Thus, the findings point to a scenario in which PA is an important factor and acts as a protective effect on body weight maintenance. In addition, the practice of PA, regardless of volume, can bring greater health benefits, 181 Gonçalves et al. Gonçalves et al. Physical activity, sitting time and BMI REFERENCES 1. Collaboration, NCD Risk Factor. Trends in adult body-mass index in 200 countries from 1975 to 2014: a pooled analysis of 1698 population-based measurement studies with 19.2 million participants. Lancet 2014;387(10026):1377–96. 2. Ministério da Saúde. Secretaria de Vigilância em Saúde. Vigilância de Fatores de Risco e Proteção para Doenças Crônicas por Inquérito Telefônico: VIGITEL 2014. Disponível em: <http://bvsms.saude.gov.br/bvs/publicacoes/vigitel_bra- sil_2014.pdf>. 3. Lee I-M, Shiroma EJ, Lobelo F, Puska P, Blair SN, Katzmarzyk PT, et al. 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https://openalex.org/W4293770958	OpenAlex	Open Science	CC-By	2,022	Do green bonds offer a diversification opportunity during COVID-19?—an empirical evidence from energy, crypto, and carbon markets	Miklesh Prasad Yadav	English	Spoken	12,368	23,157	Do Green Bonds Offer a Diversification Opportunity During COVID19? - An Empirical Evidence from Energy, Crypto, and Carbon Markets Miklesh Prasad Yadav Amity Business School Satish Kumar (  scholar.satish@gmail.com ) Malaviya National Institute of Technology https://orcid.org/0000-0001-5200-1476 Deepraj Mukherjee Kent State University Purnima Rao Fortune Institute of International Business sh Kumar (  scholar.satish@gmail.com ) alaviya National Institute of Technology https://orcid.org/0000-0001-5200-1476 Introduction Risk is inevitable in investment (Rao et al., 2021). According to Lopes (1987 ), "risk refers to situations in which a decision is made whose consequences depend on the outcomes of future events having known probabilities.". Therefore, investments require risk management, and for this, investors need to understand risk categories and sources of risk (Talluri et al., 2013). Idiosyncratic risk can be mitigated through investing in a large number of firms. However, another equally important category, i.e., systematic risk, will remain throughout the investment and give rise to volatility in returns. Therefore, Portfolio Diversification brace risk mitigation to a larger extent (Lassance et al., 2022; Zaimovic et al., 2021; Sharma et al., 2020). With the advent of globalization and digitization has dismantled the distinct separation between different financial markets. It has resulted in close interconnectedness and dependencies between the markets and their activities (Bernardi and Petrella 2015). This century has witnessed different shocks in economic crises, pandemics, wars, etc. According to International Monetary Fund (2020), climate change can lead to capital loss to economic actors and disruption in economic activities. Further supported by Guo et al. (2021), it will take away 10% of global wealth by 2050. Thus, these common global changes pose a high risk against the investors' return expectations. These events/changes have made risk interdependence more observant and visible. It further accentuates the spillover effect, a key factor in spreading the systemic risk. Hence, examining the interconnectedness among various investment alternatives is much required for the portfolio risk management. Recent literature on investment has documented the attention of investors towards green investments (Pham and Huynh, 2020), and further in this turbulent and uncertain tenure, the cryptocurrency especially bitcoin gained much attention due to its safe-haven role and potential hedging which poses tremendous opportunities for policymakers (Abakah et al., 2022). The cryptocurrency market recorded a huge market capitalization, nearly about $80 billion U.S. dollars, in October 2017 (Corbet et al., 2019). Meanwhile, a study indicates that cryptocurrency is still ambiguous to diversify in financial markets as it is considered one of the technology securities or financial bubbles (Grownwald, 2019). Assessing spillover or dynamic linkages can be a method to explore these connections. Currently, the world is interconnected because of the global integration of the financial markets and the synchronization of the business cycles (Li et al., 2021). Research Article Research Article Keywords: Green bond, energy market, cryptocurrency, carbon market, connectedness, diversification Posted Date: May 25th, 2022 DOI: https://doi.org/10.21203/rs.3.rs-1630507/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/24 Page 1/24 Page 1/24 Abstract Using the S&P green bond index (RSPGB), this study attempts to unravel the connectedness of the green bond with energy, crypto, and carbon markets. We use MAC global solar energy index (RMGS) and ISE global wind energy index (RIGW) as proxies of the energy market. In addition, we consider Bitcoin and the European energy exchange carbon index (REEX) for the cryptocurrency, and carbon market, respectively. Using the daily data from October 1, 2015, to December 13, 2021, of these constituent markets, we employ Diebold Yilmaz (2012), Barunik and Krehlik (2017), and wavelet coherence. The result reveals that the energy market (RMGS) has the highest connectedness derived from other asset classes, and bitcoin (RBTC) has the least connectedness. In addition, we note that risk transmission is heterogeneous in different scales as the short period has less connectedness than the medium and long run. Hence, the overall diversification opportunity among green bonds, energy stock, Bitcoin, and the carbon market is more in the short-run than in the medium and long-run. Surprisingly, there is no lead-lag relationship among these markets. This study provides insights to investors, policymakers, and portfolio managers. Introduction Investors need to check the dynamic linkages among various investment alternatives to benefit from a risk-return Page 2/24 trade-off between the assets having low correlation. This paper attempts to unravel the spillover from the green bond to energy, cryptocurrency, and carbon markets to check the diversification opportunity. Sustainable development is one of the most important goals that humans must achieve in the present time. Sustainable growth requires reducing carbon emissions, which is crucial for maintaining global temperature rise within acceptable boundaries that prevent climatic disasters like the disappearance of tiny islands and shore areas due to rising sea levels (Bachelet et al., 2019). These promises have concentrated on mitigation and reshaping the techniques, including green bond issuance, carbon pricing, and clean energy for tackling climate issues. Further, change in climate creates considerable portfolio investment risk, especially for the institutional investors having massive funds size for the management under them, while also presenting investment possibilities (Mercer, 2015). As a result, worldwide agreements have been reiterated to reduce CO2 emissions and limit increasing temperature levels to combat climate change (Intergovernmental Panel on Climate Change, 2014). Bitcoin is one of the leading cryptocurrencies, which raises the environmental concern as the algorithmic structure of bitcoin requires huge power backup to authenticate the transaction. In addition, the footprint (carbon) on one Bitcoin transaction is compared to 37% million tons of carbon emission (CO2) of New Zeeland (Naeem & Karim, 2021). Therefore, one asset class may affect another. Further, the issuance of green bonds is required to fund future green projects and restructure the old projects to aid in the decarbonating and moving toward a carbon-neutral economy. When the World Bank issued its first green bonds in 2008, they sparked investors' attention as a fixed-income vehicle. After that, the market of global green bonds rose from 11 billion USD in 2013 to 259 billion USD in 2019, representing an increase of 51% over 2018 (Climate Bonds Initiative, 2020). The green bond market accounts for just more than 1% of the global bond market. The majority (38%) of the revenues were employed in the power industry. It can be said that the green bond market is tiny but rising quickly. Additionally, due to the requirement of huge power backup for the algorithm of Bitcoin transactions, it is an alarming issue for the environment as it is one of the reasons for carbon emission (Naeem et al., 2021). Literature Review It has been central to examine the dynamic spillover from one market to another for portfolio diversification. Literature has shed light on dynamic spillover from the green bond to financial, commodity, crypto, energy, and other markets. We chronologically summarize these studies. There is evidence of dynamic linkage of various assets class in the studies of Tiwari et al. (2022), Liu et al. (2021); Le et al. (2021); Hammoudeh et al. (2020); Reboredo and Ugolini (2020); Garrett-Peltier (2017). Further, the results derived from these studies are mixed. Reichelt (2010) emphasized the role played by green bonds in mobilizing the private funds to mitigate climate change and argued that green bonds might be considered an initiative that confirms that financial markets can be used to support eco-friendly projects. However, public credits with AAA-rating that might be employed to channel funding to mitigate and adapt the initiative are uncommon. They further emphasized the development of fixed-income instruments that optimize the trade-off between volume and credit for mobilizing the resources on the huge scale required to combat climate change. Pham (2016) studied the connection of green bonds with other financial markets employing the multivariate GARCH model to check the possibility of dynamic linkages and found that green bond volatility is transmitted to considered financial markets. In a similar line, Reboredo (2018) found that the green bond market has a poor co-movement with the equity and energy markets which provides significant diversification benefits to investors. Unfortunately, in the corporate and treasury market, the advantages of diversification are negligible to the investors. To support this, Reboredo and Ugoline (2020) further extended their work to highlight the direct and indirect exposures of events between the green bond and financial markets by using the structural VAR. The findings imply that green bonds are tightly linked to forex and fixed- income markets. Therefore price spillovers from both markets are insignificant. Ferrer et al. (2018) employed Barunik and Krekhik's (2018) model to investigate the dynamic connectedness between the U.S. clean energy stock and crude oil prices. It was found that both have a short-term linkage only, which infers that diversification is not possible in the short run. Further, Nguyen et al. Introduction Investors need to engage in environmentally sound projects and diversify their portfolios to minimize their risk against the crypto and carbon market. This paper aims to examine the connectedness of the green bond with the energy, crypto, and carbon market. This paper investigates the dynamic connectedness of green bonds with the energy, crypto, and carbon market, collecting various indicators' daily data spanning from October 1, 2015, to December 13, 2021. The S&P green bond index (RSPGB) measures the green bond, while MAC global solar energy index (RMGS) and ISE global wind energy index (RIGW) are the proxies of the energy market. Similarly, Bitcoin and the European energy exchange carbon index (REEX) represent the cryptocurrency and carbon market. As regards Diebold and Yilmaz (2012), we document that green bond (RSPGB), energy market (RIGB), and Bitcoin (RBTC) are net receivers of shocks, and MAC global solar energy index (RMGS) dominates other markets as its transmission is high. The energy market (RMGS) has the highest risk connectedness in the short and medium run. Referring to the Barunik and Krehlic (2017) model, it is found that diversification opportunity exists in the short-run than medium and long-run as the total spillover is less in the short-run comparatively. The wavelet coherence produces a similar result as there is no coherence from the green bond to the energy market (both RMGS and RIGW) on a short and long scale. However, in the medium run (64–128 days), there is a correlation between the green bond and the energy market from the end of 2019 to mid-2020, as the red islands are present in significant areas. However, in the medium run (64–128 days), there is a correlation between the green bond and the energy market from the end of 2019 to mid-2020, as the red islands are present in significant areas. Page 3/24 Page 3/24 Additionally, the study also reports no lead-lag relationship among these markets. This research contributes to the existing studies in three folds: first, the energy market has been measured by two different proxies, namely MAC global solar energy index (RMGS) and ISE global wind energy index (RIGW), which was not included earlier in any study together. Second, more study on the spillover of a green bond is witnessed with other markets like equity, bullion, metal, cryptocurrency, carbon, and other markets. Introduction Still, very few studies are available on the spillover of green bonds with energy, crypto, and the carbon market. Third, application of novel models like Diebold Yilmaz (2012), Barunik and Krehlik (2017), and Wavelet coherence to examine the dynamic spillover among constituent markets. The remainder of this paper is structured as follows: Section 2 provides extensive literature based on the spillover of green bonds with other markets. Section 3 contains data description and econometric models. Further, section 4 furnishes empirical results and discussion, followed by the conclusion and policy implication presented in session 5. Literature Review (2021) found the spillover among S&P green bonds, MSCI global environment, Dow Jones sustainability indices, and S&P Global clean energy, which infers that risk cannot be mitigated. Similarly, Naeem et al. (2021) find asymmetric spillover among various asset classes during a different time and frequency cycles. The positive return spillover is mostly visible in the short run, whereas the negative return spillover is in both the long and short run. Further, it explains green bonds as an aid against risk in another commodity market. Further, Nguyen et al. (2021) studied the relationship between green bonds and found that green bonds act as diversification opportunities due to the low correlation effect with other assets class. A section of emerging literature furnishes green bonds as an investment alternative carrying fixed-income assets that hedge for various asset classes like renewable stocks, carbon emissions, and other markets. Sangiorgi and Schopohl (2021) surveyed asset managers on green bonds, especially in European countries. They observed that investors actively invest in the green bond market, but their preference is in those green bonds issued by sovereign countries. Additionally, it is also found that competitive pricing is one of the factors of asset managers' choice towards green bonds investment. The interest of investors is growing in a green bond, due to which the scale of growth in the green market is intriguing concerning green premium. On this note, MacAskill et al. (2020) undertook a study and encompassed whether there is "green premium" or "greenium" in green bond pricing. The consistent existence of green premium by 56% in the primary market while governments issue 70% of studies in the secondary market. We also consider the environmental impact of green bonds, which is one of the crucial components for financing green projects. In this line, Flammer (2021) strongly believes and argues the three- implication motives that companies are strengthening their image of philanthropy and being environmentally responsible by issuing and investing in green bonds. To check the hedging, Jin et al. (2020) emphasized the importance of green bonds and considered one of the effective hedging instruments for the carbon market. They computed three hedge ratios containing a battery of tests related to the transmission of dynamic linkages, for instance, symmetrical and asymmetrical dynamic conditional correlation. Literature Review (2020), by using the wavelet framework, demonstrated the time-frequency co-movement between green bonds and different assets class; they noted the presence of low and negative association of green bonds with different assets class due to which investors can park their money to mitigate the risk. Further, to investigate the role of green bonds as an effective hedging mechanism, Jin et al. (2020) considered green bonds as a diversifier for the carbon market risk. They employ DCC-GARCH, TGARCH, and GJRGARCH models and observed that the green bond index helps hedge the Page 4/24 Page 4/24 carbon futures. Similarly, academic interest is burgeoning in cryptocurrency as it plays a safe and potential hedging role in diversifying with other markets. In this line, Shahbaz and Sinha (2019), Giudici and Abu-Hashish (2019), and Baur et al. (2018) examined the connectedness between cryptocurrency (very often bitcoin) and other financial markets by employing a battery of tests and found that bitcoin is diversifier. carbon futures. Similarly, academic interest is burgeoning in cryptocurrency as it plays a safe and potential hedging role in diversifying with other markets. In this line, Shahbaz and Sinha (2019), Giudici and Abu-Hashish (2019), and Baur et al. (2018) examined the connectedness between cryptocurrency (very often bitcoin) and other financial markets by employing a battery of tests and found that bitcoin is diversifier. Various researchers analyze the dynamic spillover across green bonds and other asset classes. During the severe swings, Pham (2021) found co-movement between the green bond and equity market. Kanamura (2020) examined the price correlations between the green bond and energy commodities using daily data from November 3, 2014, to December 31, 2018. It is found that MSCI and S&P green bonds are positively connected, whereas solactive bond is negatively connected with WTI and Brent crude oil. Liu et al. (2021) undertook a study on the dynamic linkage between green bonds and the clean energy market, considering daily observation from July 5, 2011, to February 24, 2020. They employed CoVAR and found that clean energy has an asymmetrical contagion influence on green bonds. While investing in various assets class, it is noticed that a green bond is assumed to be a shield and safe-haven asset for traditional equities, stable income, commodities, and forex securities (Arif et al., 2021). In contrast, Chatziantoniou et al. To assess the factors that support the global green bond markets, such as energy prices, gold prices, and green energy stocks To enumerate the interrelationship between the carbon futures and green bond markets. Literature Review It is observed that the green bond index is a fair hedge for the carbon market, which is supported by Reboredo (2018) as he confirms the low correlation of green bonds with the energy market and finds diversification opportunities among these markets. Tiwari et al. (2022) conjecture a dynamic spillover among green bonds, renewable energy, and the carbon market. They employed TVP-VAR and LASSO dynamic connectedness based on daily observation and found that total Page 5/24 Page 5/24 connectedness amongst these assets class is heterogeneous. Additionally, it signifies that the clean energy market dominates other markets. The studies mentioned in Table 1 emphasized green finance in the form of green bonds and its drivers. Additionally, it highlights the dynamic linkages of green bonds with various asset classes like equity, energy, carbon, cryptocurrency, and other markets employing a battery of tests. To sum up, the existing studies, on the one hand, focus on the linkage or spillover from green bonds to various markets. On the other hand, a strand of literature is also found on the connectedness of green bonds with renewable energy and the carbon market. Still, there is a handful number of studies on green bond linkage with energy, carbon, and cryptocurrency market, which builds the motivation of this research. Further, the literature is fragmented and lacks the evidence regarding studying three different markets, which offer different diversification opportunities to investors. connectedness amongst these assets class is heterogeneous. Additionally, it signifies that the clean energy market dominates other markets. The studies mentioned in Table 1 emphasized green finance in the form of green bonds and its drivers. Additionally, it highlights the dynamic linkages of green bonds with various asset classes like equity, energy, carbon, cryptocurrency, and other markets employing a battery of tests. To sum up, the existing studies, on the one hand, focus on the linkage or spillover from green bonds to various markets. On the other hand, a strand of literature is also found on the connectedness of green bonds with renewable energy and the carbon market. Still, there is a handful number of studies on green bond linkage with energy, carbon, and cryptocurrency market, which builds the motivation of this research. Further, the literature is fragmented and lacks the evidence regarding studying three different markets, which offer different diversification opportunities to investors. Literature Review Page 6/24 Table 1 summarizes the recent literature published on green bonds and their interconnectedness between different markets: S.No. Paper Title Author Year Journal Purpose Findings 1 Is There an Asymmetric Relationship between Economic Policy Uncertainty, Cryptocurrencies, and Global Green Bonds? Evidence from the United States of America Syed et al. 2022 Mathematics The study investigates the asymmetric relationship between green bonds, U.S. economic policy uncertainty (EPU), and bitcoins by using the Nonlinear Autoregressive Distribution Lag It documents that green bonds are not a different asset class, and they mirror the performance of other asset classes, such as clean energy, oil prices, and bitcoins 2 The interrelationship between the carbon market and the green bonds market: Evidence from wavelet quantile- on-quantile method Ren et al. 2022 Technological Forecasting and Social change To enumerate the interrelationship between the carbon futures and green bond markets. The study discovers the positive effects of the carbon futures in the medium to long term and intermittent performance in the short term. The effects are more distinct when both markets are in a risky state. 3 Driving green bond market through energy prices, gold prices and green energy stocks: evidence from a non-linear approach Yan et al. 2022 Economic Research- Ekonomska Istraživanja To assess the factors that support the global green bond markets, such as energy prices, gold prices, and green energy stocks The findings revealed that gold and energy prices have an inferior effect on the green bonds, the green energy stocks have a swelling effect on the green bonds market 4 Dynamic nonlinear connectedness between the green bonds, clean energy, and stock price: Chai et al. 2022 Annals of operational Research To examine the dynamic nonlinear connectedness between the green bonds, clean energy, and stock price around the pandemic This study verifies the presence of nonlinear and dynamic correlation among green Page 7/24 markets: S.No. Paper Title Author Year Journal Purpose F 1 Is There an Asymmetric Relationship between Economic Policy Uncertainty, Cryptocurrencies, and Global Green Bonds? Evidence from the United States of America Syed et al. 2022 Mathematics The study investigates the asymmetric relationship between green bonds, U.S. To analyze the impact of the cryptocurrency environment attention index (ICEA) on clean energy stocks and green bonds using a range of econometric methods. Literature Review economic policy uncertainty (EPU), and bitcoins by using the Nonlinear Autoregressive Distribution Lag It t b a a a t p o c a e p b 2 The interrelationship between the carbon market and the green bonds market: Evidence from wavelet quantile- on-quantile method Ren et al. 2022 Technological Forecasting and Social change To enumerate the interrelationship between the carbon futures and green bond markets. T d p o f m lo in p t T m w m a 3 Driving green bond market through energy prices, gold prices and green energy stocks: evidence from a non-linear approach Yan et al. 2022 Economic Research- Ekonomska Istraživanja To assess the factors that support the global green bond markets, such as energy prices, gold prices, and green energy stocks T r g e h in o b g s s o b 4 Dynamic nonlinear connectedness between the green bonds, clean energy, and stock price: the impact of the COVID-19 pandemic Chai et al. 2022 Annals of operational Research To examine the dynamic nonlinear connectedness between the green bonds, clean energy, and stock price around the pandemic in the international markets T v p n d c a b e s 4 Dynamic nonlinear connectedness between the green bonds, clean energy, and stock price: the impact of the COVID-19 pandemic Chai et al. 2022 Annals of operational Research To examine the dynamic nonlinear connectedness between the green bonds, clean energy, and stock price around the pandemic in the international markets This study verifies the presence of nonlinear and dynamic correlation among green bonds, clean energy and stock prices Page 7/24 S.No. Paper Title Author Year Journal Purpose Findin 5 Impacts of COVID-19 outbreak, macroeconomic and financial stress factors on price spillovers among green bond Mensi and Rehman 2022 International Review of Financial Analysis To examine the influence of the pandemic and global risk factors on the upside and downside price spill overs of MSCI global, building, financial, industrial, and utility green bonds The s index show notew dynam volati overs streng during pande 6 Do green bonds de-risk investment in low-carbon stocks? Reboredo et al. 2022 Economic Modelling To reconnoitre how green bonds could de- risk investments in low-carbon assets by considering diverse market conditions. Literature Review The re docum that g bonds subst divers doles they a incorp low-ca invest portfo 7 Extreme directional spillovers between investor attention and green bond markets Pham and Cepni 2022 International Review of Economics and Finance To study how the spillovers between investor attention and green bond performance fluctuate across ordinary and risky market environments Spillo time-v asym and expre influe stock marke volati and e policy impro 8 Asymmetric connectedness between cryptocurrency environment attention index and green assets Kamal and Hassan 2022 The Journal of Economic Asymmetries To analyze the impact of the cryptocurrency environment attention index (ICEA) on clean energy stocks and green bonds using a range of econometric methods. Covid revea conne and c the di conta amon and a signif relatio betwe and a return S.No. Paper Title Author Year Journal Purpose Findings 5 Impacts of COVID-19 outbreak, macroeconomic and financial stress factors on price spillovers among green bond Mensi and Rehman 2022 International Review of Financial Analysis To examine the influence of the pandemic and global risk factors on the upside and downside price spill overs of MSCI global, building, financial, industrial, and utility green bonds The spill over index method shows noteworthy dynamic volatility spill overs that strengthen during the pandemic 6 Do green bonds de-risk investment in low-carbon stocks? Reboredo et al. 2022 Economic Modelling To reconnoitre how green bonds could de- risk investments in low-carbon assets by considering diverse market conditions. The research documents that green bonds have substantial diversification doles when they are incorporated in low-carbon investment portfolios 7 Extreme directional spillovers between investor attention and green bond markets Pham and Cepni 2022 International Review of Economics and Finance To study how the spillovers between investor attention and green bond performance fluctuate across ordinary and risky market environments Spillovers are time-varying, asymmetric, and expressively influenced by stock, oil, bond market volatility, and economic policy improbability. 8 Asymmetric d Kamal d 2022 The Journal f To analyze the impact f h Covid period l h h S.No. To study how the spillovers between investor attention and green bond performance fluctuate across ordinary and risky market environments Literature Review 10 Should investors include green bonds in their portfolios? Evidence for the USA and Europe Han and Li 2022 International Review of Financial Analysis To explore the role of green bonds in asset allocation Portfolios with green bonds outpace portfolios with traditional bonds wrt risk- adjusted returns. Literature Review Paper Title Author Year Journal Purpose Findings 5 Impacts of COVID-19 outbreak, macroeconomic and financial stress factors on price spillovers among green bond Mensi and Rehman 2022 International Review of Financial Analysis To examine the influence of the pandemic and global risk factors on the upside and downside price spill overs of MSCI global, building, financial, industrial, and utility green bonds The spill over index method shows noteworthy dynamic volatility spill overs that strengthen during the pandemic 6 Do green bonds de-risk investment in low-carbon stocks? Reboredo et al. 2022 Economic Modelling To reconnoitre how green bonds could de- risk investments in low-carbon assets by considering diverse market conditions. The research documents that green bonds have substantial diversification doles when To reconnoitre how green bonds could de- risk investments in low-carbon assets by considering diverse market conditions. 7 Extreme directional spillovers between investor attention and green bond markets Pham and Cepni 2022 International Review of Economics and Finance To study how the spillovers between investor attention and green bond performance fluctuate across ordinary and risky market environments Covid period reveals higher connectedness and changes in the direction of contagion among assets and a lack of significant relationship between ICEA and asset returns. Page 8/24 S.No. Paper Title Author Year Journal Purpose Findings 9 Dependence structure and dynamic connectedness between green bonds and financial markets: Fresh insights from time-frequency analysis before and during COVID-19 pandemic Elsayed et al. 2022 Energy Economics To examines the interdependence between green bonds and financial markets The interconnection between green bonds and financial markets is capricious over time. This evidence provides suggestions for international investors regarding risk management and portfolio decisions. 10 Should investors include green bonds in their portfolios? Evidence for the USA and Europe Han and Li 2022 International Review of Financial Analysis To explore the role of green bonds in asset allocation Portfolios with green bonds outpace portfolios with traditional bonds wrt risk- adjusted returns. S.No. Paper Title Author Year Journal Purpose Findings 9 Dependence structure and dynamic connectedness between green bonds and financial markets: Fresh insights from time-frequency analysis before and during COVID-19 pandemic Elsayed et al. 2022 Energy Economics To examines the interdependence between green bonds and financial markets The interconnection between green bonds and financial markets is capricious over time. This evidence provides suggestions for international investors regarding risk management and portfolio decisions. Data and preliminary analysis This study attempts to unravel the dynamic spillover of the green bond with energy, cryptocurrency, and the carbon market. To represent the indicator of the green bond, S&P green bond index (RSPGB) is considered. In contrast, MAC global solar energy index (RMGS) and ISE global wind energy index (RIGW) measure the energy market. Further, Bitcoin and the European energy exchange carbon index (REEX) are the proxies of cryptocurrency and the carbon market, respectively. These variables are considered by the following researchers, namely Hanif et al. (2021); Liu and Liu (2021); A.K Tiwari et al. (2022). The S&P Green Bond Index is designed to finance environmentally friendly projects (Tiwari et al., 2022). The MAC global solar energy index tracks the Invesco Solar exchange-traded funds (ETF) traded on the New York Stock Exchange. Further, the ISE global wind energy index tracks the company’s activity in the wind energy industry. Bitcoin is considered a decentralized digital currency that intends to follow the ideas set out in the white paper. At last, European Energy Exchange Carbon index is computed and presented as an exchange-based price for the market value of European Union emission allowances. The daily closing price of the constituent series spanning from October 1, 2015, to December 13, 2021, from Bloomberg has been collected. The main reason for collecting the data from October 2015 is that the drive toward green finance was unleashed in 2015 by the Paris Climate Agreement in the form of green investment. The focus on renewable energy and other initiatives toward green finance started (ECLAC 2017a). Further, the data is converted into log-returns using ln(Pt)-ln(Pt-1). The description of variables is presented in Table 2: Page 9/24 Page 9/24 Table 2 Data description of constituent variables Asset/index Proxy Acronyms Source Green bond S&P Green bond index RSPGB Bloomberg Energy market MAC global solar energy index RMGS ISE global wind energy index RIGW Cryptocurrency Bitcoin RBT Carbon Market European Energy Exchange Carbon index REEX Table 2 d f bl Table 2 Source: Authors' presentation Table 2 exhibits the descriptive statistics of return on the green bond, energy market, and carbon market. Considering the mean return, it is observed that the mean return of each series is positive; the mean return of Bitcoin (RBTC) and the carbon market (REEX) is high (0.0015), followed by the energy market, namely RMGS (0.0007), RIGW (0.0004) and green bond (RSPGB) that is (0.0001, therefore, the green bond is witnessed with least average return. REEX is recorded with the highest standard deviation, which depicts that the carbon market is riskier than other markets. In addition, each series is left-skewed and leptokurtic distributed, which confirms the asymmetrical pattern; the same has been confirmed by the Jarque-Bera test, which is statistically significant at a 1% significance level. This result is similar to Rbeordo (2018) and A.K. Tiwari et al. (2022). The series must be stationary to check the dynamic linkages; hence, we apply the Augmented Dickey-Fuller (ADF) and Philips and Perron (P.P.) test. The result derived from both tests infers that all series are statistically significant at a 1% level and confirm the presence of stationarity. Furthermore, these markets exhibit the ARCH effect at a 1% significant level. On this note, our choice of models to examine the dynamic linkages is supported by summary statistics. Page 10/24 Page 10/24 Table 3 Summary statistics of the green bond, energy, and carbon market RSPGB RMGS RIGW RBTC REEX Minimum -0.0241 -0.1496 -0.1259 -0.4973 -0.9787 Maximum 0.0201 0.1132 0.0989 0.2034 1.1130 Mean 0.0001 0.0007 0.0004 0.0015 0.0015 Stdev 0.0030 0.0204 0.0113 0.0423 0.0957 Skewness -0.5936 -0.5595 -1.0627 -1.0564 -0.3040 Kurtosis 7.8513 6.5817 17.7326 14.2510 73.9348 Jarque-Bera test 0.0100 0.0000* 0.0000* 0.0100* 0.0010* ADF-test 0.0010* 0.0100 0.0001* 0.0000* 0.0000* PP Test 0.0000* 0.0000* 0.0000* 0.0000* 0.0000* ARCH Test 0.0000* 0.0000* 0.0100 0.0000* 0.0000* Note: and * indicates the significance level at 1% and 0.01% respectively Figures 1 and 2 display the graphical representation of constituent series' raw and log returns, respectively. Regarding Fig. 1, the intense fluctuations has been noticed in each asset class and have the presence of a stochastic trend. Similarly, the log return of each asset under examination is more pronounced in the COVID-19 pandemic. Further, volatility clustering is seen multiple times in each series, which depicts that high and low changes follow high changes followed by low changes (Sharma et al., 2020). Econometric Models In this paper, a battery of tests like Diebold Yilmaz (2012), Barunik and Krehlik (2017), and the Wavelet coherence model have been employed to examine the dynamic spillover of green bond with energy, crypto, and carbon market. These models are time-varying and capture the spillover or connectedness in various time horizons (short, medium, and long). Investors and other stakeholders can identify the connectedness among the various markets (Antonakakis et al., 2020; Ferrer et al., 2018). A detailed understanding of these models is as below: 3.2.1 Diebold Yilmaz (2012) and Barunik and Krehlik (2017) The Diebold and Yilmaz (2012) model is based on variance decomposition analysis to check variables' magnitude and dynamic linkages (Pesaran, M.H. and Shin, Y.,1998). It helps to examine the within, to and fro transmission of information from one variable to another. In this model, the shocks of the variables can be presented through H-step forecast error variance decomposition that can be shown below: dgH ij = σ −1 jj ∑H −1 h =0 e' iΘh ∑ej 2 ∑H −1 h =0 e' iΘh ∑Θ' hei (1) ( ) ( ) Page 11/24 Page 11/24 Where dgH ij is the H-step generalized variance decomposition (GVD) matrix, ej is a selection vector, where jth element is unity and zero otherwise. Θh is the coefficient matrix in the infinite moving average representation of the non-orthogonalized VAR, Σ is the covariance matrix of the shock vector and σ −1 jj is its jth diagonal element. The generalized variance decomposition (GVD) matrix is normalized by the tow sums to obtain pairwise directional connectedness from j to i. Where dgH ij is the H-step generalized variance decomposition (GVD) matrix, ej is a selection vector, where jth element is unity and zero otherwise. Θh is the coefficient matrix in the infinite moving average representation of the non-orthogonalized VAR, Σ is the covariance matrix of the shock vector and σ −1 jj is its jth diagonal element. The generalized variance decomposition (GVD) matrix is normalized by the tow sums to obtain pairwise directional connectedness from j to i. ~ dg ij = dg ij ∑N j=1dg ij (2) In this equation, ~ Dg ijis used to calculate the generalized connectedness and ∑N j =1 ~ dg ij=1 and ∑N i,j =1 ~ dg ij = N. Econometric Models The pairwise connectedness can be shown mathematically as below ~ dg ij = dg ij ∑N j=1dg ij (2) ~ dg ij = dg ij ∑N j=1dg ij (2) In this equation, Dg ijis used to calculate the generalized connectedness and ∑N j =1dg ij=1 and ∑N i,j =1dg ij = N. The pairwise connectedness can be shown mathematically as below In this equation, Dg ijis used to calculate the generalized connectedness and ∑N j =1dg ij=1 and ∑N i,j =1dg ij = N. The pairwise connectedness can be shown mathematically as below {C}_{i⃪j}^{H}={d}_{ij}^{H} (3) where {C}_{i⃪j}^{H} denotes the cross-spillover towards i due to shock in variable j. {d}_{ij}^{H} denotes the ijth H- step forecast error variance i.e., fraction of variable i's H-step forecast error variance dur to shock in variable j. Since in general, {C}_{i⃪j}^{H}\ne {C}_{j⃪i}^{H}, there are {N}^{2}-Nseparate measures of pair-wise directional spillover. Thus, the net pairwise directional spillover is defined as: {C}_{ij}^{H}={C}_{j⃪i}^{H}-{C}_{i⃪j}^{H} (4) {C}_{ij}^{H}={C}_{j⃪i}^{H}-{C}_{i⃪j}^{H} (4) In Eq. 4, {C}_{i⃪j}^{H} denotes the cross-spillover towards i due to shock in variable j and {C}_{j⃪i}^{H} denotes the cross-spillover towards j due to shock in variable i. Next, it followed by Barunik and Krehlik (2017) model to furnish the spillover or dynamic linkages of green bond with energy, crypto and carbon market in various time horizon, for instance, short, medium and long run. This model is different from Diebold and Yilmaz (2012) model in form of time horizon (Elsayed and Yarovaya, 2019). To be precise, it decomposes the spillover obtained from D.Y. (2012) model in various frequencies based on spectral formulation of variance. This model can be expressed mathematically as follows: {\left(f\left(\omega \right)\right)}_{j,k}\equiv \frac{{{\sigma }_{kk}^{-1}{\sum }_{h=0}^{\infty }(\left\|\varPsi {\left({e}^{-iw}\right)\varSigma )}_{j,k}\right\|}^{2})}{\sum _{h=0}^{\infty }(\varPsi {\left({\omega }^{- iw}\right)\varSigma {\varPsi }^{\text{'}}\left({e}^{+iw}\right))}_{jj}} (5) In Eq. (5), \varPsi \left({e}^{-iw}\right) is considered as impulse response fourier transformation. The terminology {\left(f\left(\omega \right)\right)}_{j,k} represents jth variable band at frequency 𝜔 because of the disturbance in kth variable. Therefore, the coefficient of frequency is considered as denominator holding the jth variable spectrum, that is, the multi-spectral of the diagonal elements of the density of xi at any given frequency 𝜔. Additionally, the weighting function can be computed as follows: {\varGamma }_{j}\left(\omega \right)=\frac{(\varPsi {\left({\omega }^{-iw}\right)\varSigma {\varPsi }^{\text{'}}\left({e}^{+iw}\right))}_{jj}}{\frac{1}{2\pi }{\int }_{-\pi }^{\pi }(\varPsi {\left({\omega }^{- iw}\right)\varSigma {\varPsi }^{\text{'}}\left({e}^{+iw}\right))}_{jj}d{\lambda }^{\text{'}}} (6) Page 12/24 Page 12/24 Where, the power of the jth variable is the aggregate of frequencies to a constant value of 2𝜋. This research employed Barunik and Krehlik (2017) model to refine the result of dynamic linkages obtained from D.Y. (2012) model. It helps investors to take investment decision in form of portfolio diversification in various time frequencies. Where, the power of the jth variable is the aggregate of frequencies to a constant value of 2𝜋. This research employed Barunik and Krehlik (2017) model to refine the result of dynamic linkages obtained from D.Y. (2012) model. It helps investors to take investment decision in form of portfolio diversification in various time frequencies. Wavelet Analysis Unlike Diebold Yilmaz (2012), Barunik and Krehlik (2017) models, wavelet analysis also captures the localized power variations among constituent variables. Generally, wavelet analysis consists of the continuous wavelet transform, cross wavelet transforms, and wavelet coherence. Wavelet coherence of wavelet analysis has been applied to refine the dynamic linkages or spillvoer in various time frequencies. The wavelet coherence examines the co-movement between time series in the time-frequency domain. It helps to detect the lead-lag relationship among variables, emphasizing frequency bands and time intervals (Torrence and Webster,1999). Mathematically, the coherence is presented as follows: {R}^{2}\left(u,s\right)=\frac{{\left\|S\left({s}^{-1}{W}_{xy}\left(u,s\right)\right)\right\|}^{2}}{S\left({s}^{-1}{\left\| {W}_{x}\left(u,s\right)\right\|}^{2}\right)S\left({s}^{-1}{\left\|{W}_{y}\left(u,s\right)\right\|}^{2}\right)} (7) Where S is the smoothing operator with {0\le R}^{2}\left(u,s\right)\le 1 with value 1 showing strong co-movement between time-series and vice-versa. The value of squared wavelet coherence measures the local linear correlation between stationary time series at each scale and is analogous to the squared correlation coefficient in linear regression. Although, the wavelet squared coherence analysis only considers positive values. Thus, it does not differentiate between positive and negative directions of the relationship between the time series. Additionally, a phase difference is introduced to provide information on the direction of the relationship and the causal relationship between time series. The same can be shown as below mathematically: p y ( g ) ( ( {W}^{xy}\left(u,s\right)\right)\right\}}{\mathfrak{R}\left\{S\left({s}^{-1}{W}^{xy}\left(u,s\right)\right)\right\}} (8) where \mathfrak{I} and \mathfrak{R} are the imaginary and real part operators respectively. Graphically, the phase is represented by a black arrow on the wavelet coherence plots where a zero-phased difference implies co- movement among the series. One of the interesting facts of the model is that it represents the positive and negative connectedness with the help of arrows in graphical depiction; the left to right arrows implies the negative association while the right to the left indicates the positive association. where \mathfrak{I} and \mathfrak{R} are the imaginary and real part operators respectively. Graphically, the phase is represented by a black arrow on the wavelet coherence plots where a zero-phased difference implies co- movement among the series. One of the interesting facts of the model is that it represents the positive and negative connectedness with the help of arrows in graphical depiction; the left to right arrows implies the negative association while the right to the left indicates the positive association. Empirical Results And Discussion Table 4 reports the results obtained from Diebold Yilmaz (2012), in which the matrix's diagonal element and off- diagonal element are present within and cross-market spillovers or dynamic connectedness, respectively. Regarding the table, 'From' in the last column shows the average value of linkages or connectedness obtained from other asset classes, whereas values in the sixth row "To" represent the average value of connectedness contributed to the other markets. Page 13/24 Table 4 Results derived from Diebold Yilmaz (2012) Series RSPGB RMGS RIGW RBTC REEX FROM RSPGB 98.73 0.08 0.20 0.41 0.57 0.25 RMGS 0.75 89.35 9.52 0.24 0.14 2.13 RIGW 0.40 3.77 95.52 0.18 0.13 0.90 RBTC 0.34 0.06 0.06 99.53 0.01 0.09 REEX 0.58 0.01 0.15 0.02 99.24 0.15 TO 0.41 0.79 1.99 0.17 0.17 3.52 Net (FROM-TO) -0.16 1.34 -1.09 -0.08 0.02 Table 4 It is revealed that RMGS has the highest connectedness (2.13) derived from other assets class, followed by RIGW (0.90), while RBTC has derived least connectedness (0.09). Furthermore, RIGW is the most contributor to the other assets class (1.99), followed by RMGS (0.79); the RBTC and REEX transmit the least value of shocks to other markets. Further, net directional connectedness helps check whether an asset class transmits greater than it receives and vice versa. In a nutshell, this study reveals the net transmitter or net receiver of shocks among various assets class considered (Tiwari et al., 2022). Referring to the estimates obtained in Table 4, it is observed that green bond (RSPGB), RIGB, and RBTC are net receivers of shocks with the values − 0.16, -1.09, and − 0.08, respectively. It signifies that the MAC global solar energy index (RMGS) dominates the carbon and green market as it transmits more shocks (1.34) than other markets. Regarding the own-variable shocks depicted in Table 3, the research observed 98.73% of the green bond, 89.35% of RMGS, 95.52% of RIGW, 99.53% of RBTC, 99.24% of REEX are driven by their shocks/within the behavior. It signifies that the index movement of RMGS by network connections is greater (10.65%) comparatively. Overall, it can be seen green bond is a net receiver of spillover and integrated marginally with other markets because of its uniquely mixed features of financial resources and environmental protection. Our results differ from the study of A.K Tiwari et al. (2022) and Broadstock et al. (2020) with respect to within shock and shock by network connection. Empirical Results And Discussion Further, overall spillover, from spillover to spillover using Diebold Yilmaz (2012), has been shown in Fig. 3, respectively. In this figure, observations like 0, 500, 1000, and 1500 are equal to October 1, 2015, September 14, 2017, August 21, 2019, and October 1, 2021. December 13, 2021. The study reports that the least and highest overall connectedness is seen during the beginning of 2017 and August 2019. Further, spillover from the green bond, energy market (RMGS, RIGW), and bitcoin have a high network connection during 2019, nearly 12%. RIGW, followed by the carbon market, contributes large, considering the spillover contribution. To refine the results, next, B.K. (2017) test had been applied to examine the spillover from the green bond to constituent assets class. In addition, Barunik and Krehlic (2017) model was also applied to examine the connectedness of green bond with energy, crypto, and carbon market. The B.K. (2017) test is presented in three different frequencies like short term frequency connectedness (a day to 4 days), medium frequency connectedness (4 days to 10 days), and long- term frequency connectedness (10 days to infinity) in Table 5(I) to 5(III) respectively. In these consequent tables, "WTH" denotes within, "ABS" indicates the absolute, "From" represents the connectedness derived from other assets class, and "To" infers the spillover that has contributed to other indices. With respect to Table 5(I), it is Page 14/24 Page 14/24 observed that RMGS has the highest risk connectedness (0.90), followed by RIGW (0.69) obtained from other markets, whereas RIGW contributes high (0.80) followed by RMGS (0.60) in the short run. Moreover, RIGW, RBTS, and REEX are net contributors, while RSPGB and RMGS are net receivers of shocks in the short run. For the medium-run connectedness, RMGS is the highest receiver of shock (4.96), and RIGW is the highest contributor. The RIGW and RSPGB are net receivers, while the rest of the assets class are net receivers. Surprisingly, green bond (RSPGB) behaves differently in the medium run than short run as it is a net receiver. Regarding the long- term connectedness, RMGS remains the strong receiver, and RIGW is the highest contributor of shocks. Further, green bond (RSPGB), RIGW, and Bitcoin (RBTC) are net contributors, whereas RMGS and REEX are net receivers. Our findings are similar to studies of Hanif et al. (2021) and Liu and Liu (2021) while different from Tiwari et al. Empirical Results And Discussion (2022) as they found that green bond is not the net contributor. It is documented that green bonds are marginally integrated with the energy and carbon market in risk transmission. This risk spillover differs differently in short to long run; hence, the present research employs B.K. (2017) test in three different time horizons. Risk decomposition in different periods plays a significant role for investors, arbitragers, traders, and speculators. Of course, the linkage of the two markets becomes different because of heterogeneous responses to the shocks. In this context, the overall diversification opportunity among green bonds, energy stock, Bitcoin, and the carbon market is more in the short run than in the medium and long run as the total spillover is less in the short run; the same finding is found with the conclusion Tolliver et al. (2020). It is documented that green bonds are marginally integrated with the energy and carbon market in risk transmission. This risk spillover differs differently in short to long run; hence, the present research employs B.K. (2017) test in three different time horizons. Risk decomposition in different periods plays a significant role for investors, arbitragers, traders, and speculators. Of course, the linkage of the two markets becomes different because of heterogeneous responses to the shocks. In this context, the overall diversification opportunity among green bonds, energy stock, Bitcoin, and the carbon market is more in the short run than in the medium and long run as the total spillover is less in the short run; the same finding is found with the conclusion Tolliver et al. (2020). Table 5 (I): B.K. Test (2017) - Roughly corresponds to 1 day to 4 days (band 3.14 to 0.79) RSPGB RMGS RIGW RBTC REEX FROM_ABS FROM_WITH RSPGB 65.87 0.07 0.11 0.38 0.38 0.19 0.26 RMGS 0.22 61.03 2.71 0.21 0.14 0.66 0.90 RIGW 0.13 2.09 64.01 0.18 0.12 0.51 0.69 RBTC 0.23 0.03 0.03 75.69 0.01 0.06 0.08 REEX 0.32 0.00 0.08 0.02 90.88 0.09 0.12 TO_ABS 0.18 0.44 0.59 0.16 0.13 1.50 TO_WTH 0.25 0.60 0.80 0.22 0.18 2.05 Net 0.01 0.30 -0.11 -0.14 -0.06 Page 15/24 Page 15/24 Page 15/24 Table 5 (II): B.K. Empirical Results And Discussion Test (2017) - Roughly corresponds to 4 days to 10 days (band 0.79 to 0.31) RSPGB RMGS RIGW RBTC REEX FROM_ABS FROM_WITH RSPGB 20.18 0.00 0.06 0.02 0.12 0.04 0.24 RMGS 0.27 16.99 3.65 0.02 0.00 0.79 4.79 RIGW 0.14 1.00 18.93 0.00 0.00 0.23 1.39 RBTC 0.06 0.02 0.02 15.11 0.00 0.02 0.11 REEX 0.16 0.00 0.04 0.00 5.58 0.04 0.25 TO_ABS 0.13 0.20 0.75 0.01 0.02 1.12 TO_WTH 0.77 1.24 4.58 0.05 0.15 6.78 Net -0.53 3.55 -3.19 0.06 0.10 Table 5 (III): B.K. Test (2017) - Roughly corresponds to 10 days to inf days (band 0.31 to 0) RSPGB RMGS RIGW RBTC REEX FROM_ABS FROM_WITH RSPGB 12.68 0.00 0.03 0.01 0.07 0.02 0.22 RMGS 0.26 11.33 3.16 0.01 0.00 0.68 6.50 RIGW 0.12 0.68 12.58 0.00 0.00 0.16 1.53 RBTC 0.04 0.02 0.02 8.73 0.00 0.02 0.14 REEX 0.10 0.00 0.03 0.00 2.77 0.03 0.25 TO_ABS 0.11 0.14 0.65 0.00 0.01 0.91 TO_WTH 1.00 1.33 6.15 0.03 0.14 8.65 Net -0.78 5.17 -4.62 -0.11 0.11 Table 5 (II): B.K. Test (2017) - Roughly corresponds to 4 days to 10 days (band 0.79 to 0.31) RSPGB RMGS RIGW RBTC REEX FROM_ABS FROM_WITH RSPGB 20.18 0.00 0.06 0.02 0.12 0.04 0.24 RMGS 0.27 16.99 3.65 0.02 0.00 0.79 4.79 RIGW 0.14 1.00 18.93 0.00 0.00 0.23 1.39 RBTC 0.06 0.02 0.02 15.11 0.00 0.02 0.11 REEX 0.16 0.00 0.04 0.00 5.58 0.04 0.25 TO_ABS 0.13 0.20 0.75 0.01 0.02 1.12 TO_WTH 0.77 1.24 4.58 0.05 0.15 6.78 Net -0.53 3.55 -3.19 0.06 0.10 Table 5 (III): B.K. Test (2017) - Roughly corresponds to 10 days to inf days (band 0.31 to 0) RSPGB RMGS RIGW RBTC REEX FROM_ABS FROM_WITH RSPGB 12.68 0.00 0.03 0.01 0.07 0.02 0.22 RMGS 0.26 11.33 3.16 0.01 0.00 0.68 6.50 RIGW 0.12 0.68 12.58 0.00 0.00 0.16 1.53 RBTC 0.04 0.02 0.02 8.73 0.00 0.02 0.14 REEX 0.10 0.00 0.03 0.00 2.77 0.03 0.25 TO_ABS 0.11 0.14 0.65 0.00 0.01 0.91 TO_WTH 1.00 1.33 6.15 0.03 0.14 8.65 Net -0.78 5.17 -4.62 -0.11 0.11 To refine the connectedness of green bond with constituent assets class, wavelet coherence analysis is displayed in Fig. 4. Unconditional correlation furnishes evidence of whether the various assets class are correlated or not, which spans a long period. Still, it does not depict and explore connectedness with frequencies over time. For the same, wavelet coherence is employed. Empirical Results And Discussion It also helps to detect the lead-lag relationship among variables, emphasizing frequency bands and time intervals. The Y-axis and X-axis show the frequencies of scale and time, respectively. The time-frequency has been categorized into four cycles, namely short-scale (16–32 and 32–64 days), medium-scale (64–128 days), and long-scale (128–256 days). Similarly, the X-axis depicts the number of observations 200, 400, 600, 800, 1000, 1200, and 1400, which are on July 15, 2016, April 27, 2017, February 5, 2018, November 12, 2018, August 21, 2019, May 27, 2020, and March 3, 2021, respectively. For ease of interpretation (see Fig. 4), islands and arrows are presented in which red islands display the strong coherence (near coefficient 1) whereas blue islands show weaker coherence (Liu and Liu, 2020). To refine the connectedness of green bond with constituent assets class, wavelet coherence analysis is displayed in Fig. 4. Unconditional correlation furnishes evidence of whether the various assets class are correlated or not, which spans a long period. Still, it does not depict and explore connectedness with frequencies over time. For the same, wavelet coherence is employed. It also helps to detect the lead-lag relationship among variables, emphasizing frequency bands and time intervals. The Y-axis and X-axis show the frequencies of scale and time, respectively. The time-frequency has been categorized into four cycles, namely short-scale (16–32 and 32–64 days), medium-scale (64–128 days), and long-scale (128–256 days). Similarly, the X-axis depicts the number of observations 200, 400, 600, 800, 1000, 1200, and 1400, which are on July 15, 2016, April 27, 2017, February 5, 2018, November 12, 2018, August 21, 2019, May 27, 2020, and March 3, 2021, respectively. For ease of interpretation (see Fig. 4), islands and arrows are presented in which red islands display the strong coherence (near coefficient 1) whereas blue islands show weaker coherence (Liu and Liu, 2020). Page 16/24 Page 16/24 In the short run, there is no coherence from the green bond to the energy market (both RMGS and RIGW), crypto- market, and carbon market as the red and blue islands are scattered and not uniform. However, in the medium run (64–128 days), there is a correlation between the green bond and the energy market from the end of 2019 to mid-2020, as the red islands are present in significant areas. This might occur because of the COVID-19 outbreak; hence, investors must consider these investment alternatives in their portfolios. Conclusion And Policy Implications The new and expanding green bond market allows investors to engage in environmentally sound projects and diversify their portfolios to minimize risks (Abakah et al., 2021). In this context, one strives to seek the connectedness among various investment alternatives they have. As there are burgeoning academic interests in the environment index like carbon emission, green bond, energy market, and technological (crypto) securities, we attempt to unravel the connectedness among these markets. We consider the S&P green bond index (RSPGB) to measure the green bond to achieve our research objective. In contrast, MAC global solar energy index (RMGS) and ISE global wind energy index (RIGW) denote the energy market. In addition, to represent the cryptocurrency and carbon market, Bitcoin and the European energy exchange carbon index (REEX) are considered, respectively. We use daily data of the aforementioned markets, extending from October 1, 2015, to December 13, 2021, and apply Diebold Yilmaz (2012), Barunik and Krehlik (2017), and Wavelet coherence for the empirical purpose. Regarding dynamic spillover of green bond per se with constituent markets, the study documents various findings employing a battery of tests for the spillover like Diebold Yilmaz (2012), Barunik and Krehlik (2017), and Wavelet coherence model. It is noticed that the energy market (RMGS) has the highest connectedness (2.13) derived from other assets class, followed by another index of the energy market (RIGW) which is 0.90. In contrast, bitcoin (RBTC) has derived the least connectedness (0.09). Overall, it can be seen that the green bond is a net receiver of spillover and integrated marginally with other markets because of its uniquely mixed features of financial resources and environmental protection. Focusing on dynamic connectedness in various time scales, it is found that one index of the energy market (RMGS) is the highest receiver of shock. In contrast, another index of the energy market (RIGW) is the highest contributor in the short, medium, and long run, respectively. Additionally, the risk transmission is heterogeneous in different scales as the short period has less connectedness than the medium and long run. Hence, the overall diversification opportunity among the green bond, energy stock, Bitcoin, and carbon market is more in the short-run than in the medium and long run. Surprisingly, there is no lead-lag relationship among these markets. Empirical Results And Discussion Moving further for the long run (128–256), there is no association of green bond with the rest of the constituent assets class. Further, the movement of arrows from right to left or left to right is unclear as it is scattered and not uniform. Hence, there is no lead-lag relationship among these variables. Based on Diebold Yilmaz (2012) and wavelet coherence, the marginal integration of the green bond with the rest of the constituent variables has been reported. The movement in a variable is caused by its own shock rather than by other variables' shock. The findings are in consensus with studies of Hanif et al. (2021); Liu and Liu (2021). Conclusion And Policy Implications Badea and Pupazen (2021) have argued that bitcoin investments do not come under green investments and termed it "dirty currency." It is also governed by black swan events and can provide hedge and significant diversification potential. These findings are similar to studies by Hanif et al. (2021), Liu & Liu (2021), and Tolliver et al. (2020). Page 17/24 Page 17/24 The findings of this study proffer the various implications. First, green bonds are least volatile than rest of the constituent markets that furnishes an opportunity to investors to use green bonds as investment assets while holding other asset classes. Second, as green bonds receive most shocks from the carbon market, policymakers can consider green bonds and carbon stocks as interrelated assets class in their policy framework. Similarly, it may furnish a portfolio composition containing these stocks for portfolio diversification. Third, investors can focus on portfolio hedges containing the energy market as one of the energy markets indexes (MAC global solar energy index- RMGS) is the highest receiver of shocks. In contrast, another one (ISE global wind energy index-RIGW) is the highest contributor to shocks. Fourth, due to the less connectedness of bitcoin with other markets, investors must park their money in bitcoin as it acts as a safe haven. Additionally, our findings on the dynamic connectedness of the green bond can assist in enacting the policies that assure a smooth recovery process in various market conditions. Our findings propose that a study can be carried out to examine the dynamic connectedness employing the BEKK model, dynamic conditional correlation, Quantile VAR, TPV-VAR and asymmetrical connectedness among various markets. Based on the transmission of information, one can hold optimal portfolio weights and hedge ratios in various time duration like pre-Covid and post-Covid. Declarations Ethical Approval: We confirm that the manuscript follow compliance with ethical standards, author confirms that study does not need any approval of ethics committee. Consent to participate: “Not applicable”, manuscript does not report or involve the use of animal or human data or tissue. Consent to participate: “Not applicable”, manuscript does not report or involve the use of animal or human data or tissue. Consent to publish: “Not applicable”, manuscript does not contain data from any individual person. onsent to publish: “Not applicable”, manuscript does not contain data from any in Consent to publish: “Not applicable”, manuscript does not contain data from any individual person. Author contributions: Miklesh Prasad Yadav: conceptualisation, acquisition of data, analysis and interpretation, original draft writing; Satish Kumar: original draft editing, visualization, supervision, analyze and interpret; Deepraj Mukherjee: supervision, conceptualisation, Table formulation and editing, data validation; Purnima Rao: data compilation, writing, Data conversion and paper review. All authors contributed to the article and approved the submitted version. Funding: No funding received for this manuscript. Competing Interest: The authors declare no competing interests. Data availability: All data generated or analyzed during this study can be available from corresponding author on request. References Research Letters 25: 103–110. https://doiorg/101016/jfrl201710012 8. Bernardi M, Petrella L (2015) Interconnected risk contributions: A heavy-tail approach to analyze US financial sectors. 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J Clim 12:2679–2690. https://doi.org/10.1175/1520-0442(1999)012%3C2679. :ICITEM%3E2.0.CO;2 55. Yan L, Wang H, Athari SA, Atif F (2022) Driving green bond market through energy prices, gold prices and green energy stocks: evidence from a non-linear approach. Economic Research-Ekonomska Istraživanja: 1– 21. https://doiorg/101080/1331677X20222049977 56. Zaimovic A, Omanovic A, Arnaut-Berilo A (2021) How Many Stocks Are Sufficient for Equity Portfolio Diversification? A Review of the Literature. Journal of Risk and Financial Management 14(11): 551. https://doiorg/103390/jrfm14110551 Page 22/24 Figure 1 Time series plot of raw series Time series plot of raw series Page 22/24 Figure 2 Time series plot of constituent return series Figure 3 Figure 2 Time series plot of constituent return series Figure 2 Time series plot of constituent return series Time series plot of constituent return series Page 23/24 Figure 3 Figure 3 Figure 4 Wavelet Coherence Analysis Page 24/24	45,672
hal-03609880-Geophysical%20Research%20Letters%20-%2015%20November%201995%20-%20Echalar%20-%20Aerosol%20emissions%20by%20tropical%20forest%20and%20savanna%20biomass.txt_1	French-Science-Pile	Open Science	Various open science	2,022	Aerosol emissions by tropical forest and savanna biomass burning: characteristic trace elements and fluxes. Geophysical Research Letters, 1995, 22 (22), pp.3039-3042. ⟨10.1029/95GL03170⟩. ⟨hal-03609880⟩	None	English	Spoken	2,634	6,441	Aerosol emissions by tropical forest and savanna biomass burning: characteristic trace elements and fluxes F. Echalar, A. Gaudichet, H. Cachier, P. Artaxo 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 privé Aerosol emissions by tropical forest and savanna biomass burning: characteristic trace elementsand fluxes F. E chalar and A. Gaudichet LISA- URA CNRS 1404, Universit6sParis 7 et Paris 12, Cr6teil,France H. Cachier CentredesFaiblesRadioaetivit•s,LaboratoireMixte CNRS - CEA, Gif-sur-Yvette,France Institutode Fisiea,Universidadede S•o Paulo,S•o Paulo,Brazil Abstract. Aerosolparticles werecollected duringthe dry significant amountof traceelementsinto the atmosphere may alsobe importantas a pathwayto exportingnutrients seasonin the amazonian forest and in African and Brazilian savannas. Samplingwas performedinsidethe plumesof vegetation fires and in the local background atmosphere. Samples wereanalysed for theirelemental composition by Particle InducedX-Ray Emission (PIXE). Enrichment from the sourceregion. Previousworkson traceelementemissionby fires in the tropicalbelt are scarce.Ward et al. [1991 and 1992] present the elementalcontentof particlesemittedby rain forestand cerradofires in Brazil. Artaxoet al. [1993] factorsrelative to the local backgroundwere calculated. Savannafire aerosolsare characterised by enrichments in present thetraceelement content of particles obtained by elements like K, P, CI, Zn, and Br, whereas forest fire airbornesamplingof fire plumesin the Amazonbasin. Quantitative estimates of the intensityof emissions in the emissions areenrichedin Si andCa. The unexpected low potassiumenrichmentfactor for the forest fires could be tropical belt are still scarcer: besidesthe estimate of due to the prevailingsmolderingconditions,whereasthe potassium emission by.firesin theAmazon basin[Andreae, highvaluesof potassium enrichment factorduringsavanna fires couldbe associated with the predominant flaming conditions. This resultindicates that potassium may be considered a goodtracerof theflamingphaseof firesonly. 1988],to ourknowledge only onedatasetis available:that obtainedfrom our ground-based measurements conducted in IvoryCoast[Gaudichet et al., 1992]. The main objective of this work is to characteriseand compare trace element emission from fires of three different types of savannasand from the south-western Emissionfactorsof P, S, C1, K, Ca, Mn, Zn, Br and total particulatematterwere calculatedfor African savannafires. Our estimatesof the annualpotassium and zinc emissions amazonian rainforest.Theaimof thisstudyis to verifyif thereis a fingerprint thatmaycharacterise savanna firesor, in a moregeneral scale,tropicalbiomass burning. We will by tropical savannafires indicatethat the contributionof this source should be taken into account to understandthe biogeochemical cycleof theseelements. also estimatethe intensitiesof the trace element emissions andtry to assess the strength of the biomass burning Introduction SOurCe. Biomassburningemissionof gasesand particlesto the atmosphere attractsnowadaysmuchattentiondue to their possible effectson the chemistryof the atmosphere andon climate[Cmtzenand Andreae,1990]. Biomassburning emissionsare concentratedin the tropical belt, where is consum morethan 80% of biomassburntin the world, a half of it duringsavannafires [Andreae,1991]. Biomass burningmight representalso a significantsourceof trace elements thatwouldhaveto be considered for the studyof their global biogeochemical cycles. Indeed, the global extension of biomassburningactivitiesresultsin a large amountof gasesandparticlesinjectedintotheatmosphere, andtraceelementspresentin the bulk plant materialor in particlespreviouslydeposited on the vegetation may be foundin theseproducts. On a localscale,the injectionof a ExperimentalSitesand Methods Samples werecollected at fourecologically differentsites: (a) the guineansavanna:in Lamto, Ivory Coast.The vegetation at this siteis representative of West Africadense humidsavannas (abovegroundbiomass densityof about9 ton/ha[Lacauxet al., 1993]);thesesamples werecollected inJanuary 1991during theFOS/DECAFE experiment; (b) the southAfricanbush:il• the KrugerNationalPark, located in theNE partof SouthAfricaandconstituted by verydrylowdensity savannas (1.4to 5 ton/ha[Trollope et al., 1993]); (c) the Brazilian savanna, or "cerrado": in the IBGE (Brazilian Institute of Geography andStatistics) Ecological Reserve, located 35 kmsouthof Brasilia;onegrassland and twowoodedsavanna plotswereburnt(typicaldensities of 7 to 10ton/ha[Ka•an et al., 1994]); (d) in thesouth-western partof theAmazonian rainforest, Copyright 1995bytheAmerican Geophysical Union. about 100 km south of Porto Velho, in the State of Rond6nia (about360 ton/ha[Kauffman et al., personal Papernumber95GL03170 communication]). 0094-8534/95/95GL-03170503.00 While in the savanna plotsfire was alwaysset to the 3039 3040 ECHALAR ET AL.: AEROSOLS FROM TROPICAL FOREST AND SAVANNA FIRES standing vegetation, in rain forestplotsmostof treeshad beencUtdown40to 60daysbefore thefire,otherwise they wouldnothavebeendryenough to burnefficiently. An importantdifference that mustbe pointedoUtis that mostof the savanna vegetation burnsquicklyduringthe flamingphase,whilein the foresttrunksmayremainon fieldandkeepsmoldering or glowing several daysafterthe fire. According to Wardet al. [1992],morethan97 % of carbon in cerrado firesis released during theflaming phase, whilein forestfiresmostof biomass is consumed during thesmoldering phase.In thefollowingdiscussion all results concerning savanna sreferto samplescollectedunder flamingconditions, whileresults concerning tropicalforest fires refer only to samplescollectedin the individual plumesof smoldering or glowingtrunks. z z[.• __.• ma dt•.__!n I A! Si P S C! K Ca Ti V Cr Mn Ni Cu Zn Br / • S.Africa • I.Coast•q• Brazil s0 • 70 (b) Forest fir•• • Z 50 • • 20 Z c The samplingandanalyses protocolsare described in detail in Gaudichet et al. [1992].In short,aerosol sampling was carried oUt in biomassburningplumes using an aUtonomous andmobilepumping unit.Thissampling set-up allowedthe simultaneous collection of different typesof samples in parallel,in the fire plumesor in background conditions. Parallelto the aerosolsampling,CO2,in Ivory Coast,and CO2 and CO, in South Africa, were measuredin real time usinga portableinfraredspectrometer (BINOS analyser),in orderto obtaina quantitativeestimateof the intensityof aerosol emissions. Traceelementconcentrations wereobtainedusingthe PIXE method [Johanssonand Campbell, 1990]. Filters were weighedtwice, with a Mettier M3 electronicbalance, beforeandaRerthe samplingto obtainthe particulatemass of the samples. Parallelto some of our trace elementssamples,particles were collected on Whatman GF/F glassfiber filters for black carbonanalysisby coulometrictitration [Cachieret al., 1993]. Similar measurements were carriedoUt in the background atmosphere of the tree savannasites. Results and Discussions Characteristic Trace Elements Particlessampledin plumesof savannaandforestfiresmay havetwo differentsources:the combustionof vegetationor the remobilisation of particles that were previously depositedon the vegetation.Trace elementsthat may characterisebiomassburningshouldbe thosefor which the combustion of the vegetation iõthemainsource. To determinewhich are theseelementsourfirst stepwasto assesthe characteristicsof the particlesdepositedon the vegetation.The trace element content measured in the background aerosol is supposed to reflect these characteristics. In the Ivory Coast,the situationwassimple: theatmospheric aerosolis composed of particlesemittedby savanna fires and by soil dust[Gaudichetet al., 1992]. BUt in the othersitessourceapportionment wasmorecomplex. Our approachthen was to use enrichmentfactorsof each elementin the plume with referenceto its concentration in the local background.Iron, which is assumednot to be predominantly produced by the combustion of the vegetation,was usedhere as the referenceelement.Median valuesof theseenrichmentfactorsare shownin Figure1. ß • A! Si'P S'C!'K 'Ca' •'V 'Cr'Mn'Ni •. Cu Z Brazil Figure 1.M•ian trace elem•t•c••t favors for(a) savanna and•) fore• f•e aerosols: •e)in the plume divid• by •e)in the local bac•round• Back•o•d aerosol data for •e amazonian fore• ob•• •om •xo et al. [1988]. Fore• samplesco•e•ond to smolder•g conditions, •e •va•a samples co•e•ond to fiam•g conditions. Elem•ts •derl• are •ch• and have ratiosto Fe • •e plme •ti•ically d•er•t •om •e •me elem•tal ratios • •e back•o•d atmo•here accord•gto Stud•t's t-te• at •e 95% conduce level. For the three types of savannaa similar patternmay be observedin this figure: biomassburningparticlesare very enrichedin K, CI, Br, andZn (enrichmentfactor> 10) and moderatelyenrichedin P, Ca, and Mn (4 < enrichment factor < 10). All these elementsare presentin plants [Hewitt, 1983] and may be associated to the combustionof vegetation.By contrast,biomassburningparticlespresent low enrichmentfactorsfor Al and Si, indicatingthat the contribUtionof the combustion of vegetation to the measuredconcentrationis small. It must be pointedoUt that low enrichment factors do not mean low concentrations. In guineansavannasamples,for example, Si, Al and Fe concentrationlevels are very high, the low enrichmentfactors indicating that the fire induced an intenseremobilisation of terrigeneous particlesdeposedon the vegetation[Gaudichetet al.; 1992]. There are some differencesregardingsomeminor elements:V is enriched in cerradoand south-AfiScansavannasplumes,while Cr and Ni are enrichedmostly in African savannaplumes. Theseenrichmentsare somewhatsurprising,as they are elementsusuallyassociatedwith industrialpollUtion.The re-entrainement of industrialparticlescannot be excluded, bUt,astheseelementsmay be presentin the plants[I-Iewitt, 1983],a "biomassburning"originmustalsobe considered. In theforestthe patternis different:aerosolsare enrichedin Ca, Si. The unexpectedlow enrichmentin K, C1, Zn and P of forestfire aerosolsmay be explainedby the intensityof natural emissionsfrom the vegetation.These emissions dominatethe backgroundaerosoland are themselvesrich in suchelements[Artaxo et al., 1988]. This resultsuggest 1.Excess potassium to sootcarbon ratiosinthefireplumes whereA arethe abovebackground concentrations andk is a andinthebackground atmosphere. indicatingthe proportionof fuel carbon to the measuredconcentration correctedfor the crustalcontribution. released as CO2. Typeof fire K/Cs guinean savanna fires 0.64ñ0.28 S. Africansavanna fires cerradofixes forestfires 1.14ñ0.96 0.52A:0.61 0.055ñ0.039 wood smoke aged plumes offorest firesin 0.007ñ 0.002 0.11 Amazonia In the case of African savannafires it was possibleto calculateemissionfactorsfor everysample(Table 2), using ourCO andCO2data,carboncontentin ashesandpartially burnt material [T. Kuhlbusch et al., unpublished Cachier etal.[1993] thiswork manuscript],and literaturedata on the emissionof other thiswork carbonaceous products[Lobertet al., 1991; Bonsanget al. thiswork 1993].Takingaccountof the emissionfactorsvariabilityat Curtie[1991] eachsamplingsite, it may be considered that there is little Reference Andreae etal.[1988] difference ml I thatSi or Ca mightbe considered bettermarkersfor wood fire thanpotassium, whichis commonly presented as an ubiquitous tracerof biomass burning aerosol. However,due to the wide variety of Si and Ca sources,their role as tracersis questionable. potassium to blackcarbonratio(Table1), whichis often usedasa tracerof biomass burning aerosols [Cachier et al., factors obtained at two fire thatis actuallyburnt,i.e., the burningefficiency. Global fluxes from tropical savannafires were computed assumingthat the amountof biomassconsumedeachyear is 3700 tons of dry matter [Andreae,1991]. These results (Table2) indicatethat pyrogenicemissionsmighthave an impacton the globalcycleof sometraceelements.For Zn, 1993].Thisradiovariesin therange0.007- 1.14,showing lowestvaluesfor woodburning.Ourforestresultsarethen in agreement with previousmeasurements. The K contentin particulateemissionsfrom savannafires smallervariation(minimum%f 0.16+0.30% in the forest emission its emissionfactor, the biomassdensityexposedto fire, the surfaceburnt, and the fractionof biomassexposedto the In orderto checkour puzzlingK results,we introducethe (3.2to 4.4%) isabout10timeshigher thanin oursamples fromforest fires(0.3+ 0.1%). It isnecessary to verifyif this differenceis relatedto a differentvegetation composition or to differences in theprevailing fire stage (flaming for savannas versus smoldering for the forest). Potassium contents of the vegetation matrixpresenta between different ecosystems. Consequently they may be characteristic of tropicalsavannafires and may be usedin globalestimates. To getanideaof the importance of the atmospheric transfer of trace elementsby biomassburning,somecomparisons with otherlocal or regionalsourcesmay be proposed. flux of a givenelementmay be estimatedas the productof the main sourceis anthropogenic:savannafire emissions could correspondto about 6 % of all anthropogenic emissions and about 20% of the total natural emissions (131 x 103ton/year[NriaguandPacyna,1988];and 45 x Table 2. Median andrangesof emissionfactorsfrom african savannas fires,andtheannualfluxemittedto theatmosphere bythe [Kauffman et al.;personal communication] to about0.60% fires in all tropical savannas. Fluxes (in Tg/year) were estimated in cerrado[Kauffinanet al.; 1994]), so the observed usingthe meanof the two medianemissionfactors,andthe amount differences in K aerosol contents maythenbe attributed to burnteachyearin thetropicalsavannas (3690Tg/year thedifferent combustion processes: flamingx smoldering. ofbiomass Ward et al. [1992] in another amazonianforest site observed alsothatpotassium content in the smoldering [Andreae,1991]).In parentheses thenumberof pointsfor whichthe emission factors were calculated. phaseis lowerthanin theflamingphase.We suggest that the thermodynamic conditions of the smoldering phase hinders thevolatilisation ofpotassium; it maybesupposed alsothatmostofthepotassium hasbeenusedupduring the flamingphaseof the forestfires.However, smoldering P resultsfrom Ward et al. [1992]are nearlyone orderof magnitude higherthanthoseobserved in oursamples. This difference mightbe explained by thefactthatWardet al.'s smoldering sampleshavebeencollected just after the flamingphase,andcouldhaveintegrated the plumesof phase,whereas our samples havebeencollectedlater,in SourceStrength and Flux Estimates Quant itative assessment s rely on estimatesof emission S FLUX (Tg/year) 0.011 t (12) 0.029 0.0047 (14) C! Ca TPM 0.020 t (12) o.22 t (12) 0.30(17) 0.34 t (12) 0.16 - 0.61 0.067- 0.013 (17) 0.062 0.0013(16) 0.0039 0.0012 (16) 0.00055- 0.0047 0.0026 (12) 0.0015- 0.0057 0.030 t (12) 0.0030- 0.005 0.008 0.0089 0.0045 (4) 0.0019- 0.080 0.0033 0.0032 t (11) 0.00036- 1.2 0.15 0.0015(4) 0.00018- 0.92 1.0 0.013 0.017 6.5(17) 7.9t (4) 2.9- 16 1.2- 16 tfrom Gaudichet etal.[1992]. 0.13 0.0030 - 0.050 0.057 - 1.2 followingequation: EFx = k x AX / ACO2 0.050 0.28(17) 0.00043- Zn 0.12 • 0020- 0.11 - 0..50 0.0056- Mn 0.017 0.052 (17) 0.0 28 - factor.Theemission factorof a givenelement represents Br theamount of thiselement emitted to theatmosphere per unit massof vegetationconsumed by the fire. The computation of emission factors wascarriedoutusingthe Ivory Coast (g/kgdrymatter) 0.00052- smoldering vegetation andagedproducts fromtheflaming K theindividual plumesof smoldering. SouthAfrica (g/kgdrymatter) 26 3042 ECHALAR ET AL.: AEROSOLS FROM TROPICAL FOREST AND SAVANNA FIRES 103ton/year[Nriagu,1989]respectively). On a regionalscalethe emissionof trace elementsto the atmosphere mightbe compar•leto lossof traceelements by run-off. This is the case of West Africa, where we estimatedthe releaseof K to the atmosphere by savanna fires to be about2.25 kg/ha/year,which is comparableto the lossof potassiumby run off (1.4 kg/ha/year[Lesacket a!., 1984]). Thus, in this region the emissionto the atmosphere by the faresmay play an importantrole and mustbe considered for the K cycle. Acknowledgements. This work was part of the DECAFE program,funded by the French CNRS and the Nfmist•rede l•Environment. The samplesfromthe SouthAfricansavannafires were eelleered in the frame of the IGAC/BIBF, X/SAFARI-92 experiment. P. Artaxe acknowledges the financialsupportfrom FAPESP, grant 90/2950-2. Technicaland administrativehelp fromthe KrugerNationalPark staff,especiallyA.Potgieter,from the [BGE and [BAMA staffs,and from J.G. Goldammer, L. and W. Trollope, H. Luearelli and B. Dias is gratefully acknowledged. We are indebtedto studentsfrom Potsehestrom l.h•iversity(South Africa), Freiburg University and the Max PlanckInstitutefor Chemistry(Germany),andthe Universityof S•o Paulo(Brazil)for theirhelpduringfire plumesampling. We thankthe SouthAfricangovemmentandthe S. AfricanNational Park Servicefor permissionto conductthis research. References Hariss, G.F. Hill, D.J. Jacob, M.C. Poreira, G.W. Sachse,A. W. Setzer, P.L. Silva Dias, R.W. Talbot, A.L. Torres, and S.C. Wofsy, Biomass Buming Emissionsand Associated Haze Layersover Amazonia,J. Geephys.Res., 93D2, 15091527, 1988. Andreae, M. O., Biomass Buming: Its History, Use and Distributionand Its Impacton Environmental Quality and GlobalClimate in Global BiomassBurning: Atmospheric, Climaticand BiosphericImplications,editedby J. S. Levine, pp.3-21, The Mrr Press,Cambrigde, Mass.,1991. Artaxe, P., H. Storms,F. Bruynseels,R. Van Grieken,and W. Maenhaut,Compositionand Sourcesof Aerosolsfrom the AmazonBasin,d. Geephys.Res., 93D2, 1605-1615, 1988. Artaxe, P., M. Yamasoe, J.V. Martins, S. Koeinas, and S. Carvalho, Case Study of AtmosphericMeasurementsin Aerosol Uptakeand UtilisationbyPlants,editedby D. A. Robband W. S. Pierpoint,AcademicPress,London,1983. Johansson,S. A. E., and J. L. Campbell,PIXE: A Novel Technique for ElementalAnat3,sis, JohnWiley& Sons,New York, 1988. Kauffman, J. B., D. L. Cummings, and D. E. Ward, Relationships of Fire, BiomassandNutrientDynamicsalong a Vegetation Gradientin the BrazilianCorrado,J. Ecology, 82, 519-531, 1994. Lacaux, J.-P., J.-M. Brustot, R. Delmas, J.-C. Menaut, L. Abbadie,B. Bensang,H. Cachier,J. Baudot,M. O. Andteac and G. Helas, Biomassbumingin the tropicalsavannasof Ivory Coast:An overviewof the field experimentFire of Savannas(FOS/DECAFE91), Y. Atmos.Chem.,in press, 1993. Lesaek, L., R. Hooky, and J. Melaek, Transportof earben, nitrogen,phosphorus andmajorsolutesin the GambiaRiver, West Africa,Limnol. Oceanogr.,29, 816-830, 1984. Loberr, J.M., D.H. Seharffe, W.M. Hao, T.A. Kuhlbuseh,R. Souwen, P. Wameek, and P.J. Crutzen, Experimental Evaluationof Biomass Buming Emissions:Nitrogen and CarbonContaining Compounds, in GlobalBiomass Burning: Atmospheric, Climaticand BiosphericImplications,edited by J.S.Levine,The M[T Press,Cambridge, Mass.,289-304, 1991. Andreae,M.O., E.V. Browell,M. Garstang,G.L. Gregory,R.C. Brazil: Maenhaut, Trace Elements in Tropical African Savanna BiomassBumingAerosols,J. Atmos.Chem.,in press,1992 Hewitt,E. J., A Perspective of Mineral Nutrition:Essentialand Functional Metals in Plants, in Metals and Micronutrients Emissions in Amazon Basin Biomass Buming, in Fire in the Environment: The Ecological, Atmosphericand Climatic Importanceof VegetationFires, editedby P.J. Crutzen and J.G. Goldammer,John Wiley & Sons, New York, 1993. Bensang,B., U. Basslet C. Boissard,M.F. LeCloaree,and J.C. Menaut,Methane,CarbonMonoxideandLightnon-Methane HydrocarbonEmissionsfrom African SavannaBumings,J. Atmos.Chem.,in press,1993. Caehier, H., C. Liousse, P. Buat-M•nard, and A. Gaudiehet, Particulate Content of Savanna Fire Emissions, d. Atmos. Chore.,in press,1993. Cnatzen, P.J., and M.O. Andteac, Biomass Buming in the Tropics: Impact on Atmospheric Chemistry and Biogeochemieal Cycles,Science,250, 1669-1678,1990. Currio, L.A., 1991, Source apportionmentof atmospheric particles,IUPAC: Commissionon EnvironmentalAnalytical Chemistry. Gaudiehet, A., F. Eehalar, B. Chatenet, J.P. Quisefit, G. Malingre, H. Caehier, P. Buat-M•nard, P. Artaxe, and W. Nriagu,J. O., and J. M. Paeyna,QuantitativeAssessment of WorldwideContaminationof Air, Water and Softsby Trace Metals, Nature, 333, 134-139, 1988. Nriagu,J. O., A Global Assessment of Natural Sourcesof Atmospheric TraceMetals,Nature,338, 47•49, 19119. Trollope,W.S.W., A.L.F. Potgieter,N. Zambatis,and L.A. Trollope,SAFARi92:Charaeterisadon of Biomassand Fire Behaviourin ControlledBurnsin the KnsgerNationalPark, SAFARI (SouthAfrica Fire-Atmosphere ResearchInitiative) Workshop, Stellenboseh, SouthAfrica,26-29May, 1993. Ward, D. E., A. W. Setzer, Y. J. Kaufman, and R. A. Rasmussen, Characteristics of Smoke Emissions from BiomassFiresof the AmazonRegion- BASE-A Experiment, in Global BiomassBurning: Atmospheric,Climatic and BiosphericImplications,editedby J. S. Levine,The Mrr Press,Cambrigde,Mass., 199I. Ward, D. E., R. A. Susott,J. B. Kauffman,R. E. Babbitt,D. L. Cronmings, B. Dias, B. N. Holben, Y. J. Kaufman,R. A. Rasmussen, and A. W. Setzer, Smoke and Fire Characteristics for Cerrado and Deforestation Bums in Brazil:BASE-BExperiment,J. Geephys.Res., 97D, 1460114619, 1992. F. Eehalar and A. Gaudiehet, LISA URA CNRS 1404, Facult• de Sciences,61, Av. du Gal. de Gaulle, 94010, CrYtoil Codex, France (e-mail: echalar(•univ-paris 12.fr and gaudichet(•univ-pads 12.fr). H. Caehier, Centre des Faibles Radioaetivit•s,CNRS-CEA, Av. de la Terrasse,91198, Gif -sur-Y vette,France ( e-mail: eaehier(•eole.cfr.enrs-gif. fr). P. Artaxe,Institutede Fisiea- USP,CaixaPostal20516,eep 01452-990,S•o Paulo-SP , Brazil (e- mail : artaxo (• ifusp .br). (Received 10/21/94 ; ; .).	40,384
W2034054059.txt_1	Open-Science-Pile	Open Science	Various open science	2,014	Sensing light and sound velocities with phoxonic crystals	None	English	Spoken	4,422	7,482	Sensing light and sound velocities of liquids in two-dimensional phoxonic crystals Samira Amoudache, Rayisa Moiseyenko, Yan Pennec, Bahram Djafari-Rouhani, Antoine Khater, Ralf Lucklum, Rachid Tigrine To cite this version: Samira Amoudache, Rayisa Moiseyenko, Yan Pennec, Bahram Djafari-Rouhani, Antoine Khater, et al.. Sensing light and sound velocities of liquids in two-dimensional phoxonic crystals. SPIE Smart Structures/NDE 2014, Conference 9062 - Smart Sensor Phenomena, Technology, Networks, and Systems Integration VII, 2014, San Diego, United States. 90620S, 8 p., �10.1117/12.2044855�. �hal-00978927� HAL Id: hal-00978927 https://hal.science/hal-00978927 Submitted on 14 Sep 2021 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 Sensing light and sound velocities with phoxonic crystal Samira Amoudachea, b, Rayisa Moiseyenkoa, Yan Penneca, Bahram Djafari Rouhania, Antoine Khaterc, Ralf Lucklumd, and Rachid Tigrineb a Institut d’Electronique, de Microélectronique et de Nanotechnologie, UMR CNRS 8520, Université Lille 1, 59655 Villeneuve d’Ascq, France b Laboratoire de Physique et Chimie Quantique, Université Mouloud Mammeri, B.P. 17 RP, 15000 Tizi-Ouzou, Algérie c Laboratoire de Physique de l’Etat Condensé, Université du Maine, 72085 Le Mans, France d Institute of Micro and Sensor Systems (IMOS), Otto-von-Guericke-University, Magdeburg, Germany ABSTRACT We study theoretically the potentiality of dual phononic-photonic (the so-called phoxonic) crystals for liquid sensing applications. We investigate the existence of well-defined features (peaks or dips) in the transmission spectra of acoustic and optical waves and estimate their sensitivity to the sound and light velocities of the liquid environment. Two different sensors are investigated. In the first one, we study the in-plane transmission through a two-dimensional (2D) crystal made of cylindrical holes in a Si substrate where one row of holes is filled with a liquid. In the second one, the out of plane propagation is investigated when considering the transmission of the incident wave perpendicular to a periodic array of holes in a slab. Such ultra compact structure is shown to be a label-free, affinity-based acoustic and optical nanosensor, useful for biosensing applications in which the amount of analyte can be often limited. *yan.pennec@univ-lille1.fr Keywords: Phononic, Photonic Crystal Sensor, Simulations, Confinement, Resonances 1. INTRODUCTION Photonic crystals [1, 2] and their acoustic counterpart, the so-called phononic crystals [3] are now well-known for their ability to guide, control, and manipulate the propagation of the optic and acoustic waves. These properties are mainly related to the possibility of band gaps in their band structure that allow the existence of localized modes and confined optic/acoustic waves. Moreover, during the past few years, there has been an increasing interest towards structures exhibiting simultaneous phononic and photonic band gaps, the so-called phoxonic crystals, thus allowing dual confinement of phonons and photons [4, 5]. From the point of view of sensing applications, several papers have already shown the capability of photonic crystals for detecting small variations in the refractive index of gases and liquids and have opened the way to a platform for a new class of sensors. In general, the sensing phenomena is based on the high sensitivity of localized modes (associated with defects) appearing inside the gaps of the photonic crystals to the variation of the index of refraction in the surrounding fluid [6-8]. In a recent work [9], some authors have also studied the normal transmission through a slab perforated periodically with holes and measured the shift of a well-defined feature in the spectrum with the refractive index of the embedding liquid. In contrast, phononic crystals have only been recently proposed as a possible platform for the investigation of the acoustic velocity of a liquid filling the hollow parts of the structure [10, 11]. Thus, the potentiality of different geometries of phononic crystals for sensing applications still needs several further investigations. Moreover, some of the structures may be suitable for a dual measurement of both acoustic and optical velocities of the analyte. The general background is the great deal of works we have devoted during the last few years to the engineering of the dispersion curves and band gaps in phononic [3] and phoxonic [4, 12] crystal slabs. To make a phononic/photonic sensor, one needs to design a structure in which the transmission coefficient displays welldefined features that are very sensitive to the acoustic/optic velocity of the infiltrating liquid. These features should be relatively isolated in frequency in order to allow the sensing of the probed parameter on a sufficiently broad range. In 1 this paper, we investigate theoretically the transmission spectra in two geometries of periodic crystals and discuss the physical origin of the peaks and dips in the spectra and their usefulness for the sensing applications. Two configurations are considered for the propagation of the incident waves with respect to the phoxonic crystal. In the case of in-plane propagation, we study the transmission coefficient through a two-dimensional (2D) crystal made of cylindrical holes in a silicon substrate where one row of holes is filled with a liquid. In the case of out-of plane configuration, we study the transmission of the incident wave normally to a slab perforated periodically with holes. The FDTD approach has been used to resolve the photonic and phononic wave propagation. This method solves either the electromagnetic or the elastic wave equations by discretizing time and space and by replacing derivatives by finite differences in the equations of motion. As compared to other numerical method such as plane wave expansion (PWE), it presents the advantage of allowing simulation of mixed phononic structures, composed of both fluids and solids. The results for the two incident configurations are respectively presented in sections 2 and 3. Some conclusions are drawn in section 4. 2. IN-PLANE PROPAGATION Figure 1a represents a three dimensional (3D) schematic view of the investigated periodic structure composed of a silicon substrate, drilled with infinite air cylinders arranged in a square array of lattice parameter a. Figure 1b represents the elementary unit cell used for the calculation. As seen in figure 1(a), all cylinders of radius r1 are filled with air except one row of different radius r2 in the middle of the structure which contains the fluid to be probed. In all the following calculations, the normalized radius of the inclusions constituting the perfect structure is fixed to r1/a = 0.25 while r2/a is chosen variable for the optimization of the sensor. (a) (b) PCC PM.L PCC Y Figure 1: (a) 3D-schematic representation of the phoxonic crystal containing one row of liquid filled cylinders. (b) Elementary unit cell used for the calculation of the transmission of optical and acoustic waves. On each y-boundaries of the unit cell perfect matching layers (PML) are applied while on the x-boundaries periodic boundary conditions (PBC) insure the periodicity of the crystal. The defect line of holes have a radius r2 which can be different from the radius r1 of the phoxonic crystal of lattice parameter a. To study the acoustic waves silicon is considered as a cubic material with the elastic constants C11 = 16.57 1010 N/m2, C44 = 7.962 1010 N/m2, C12 = 6.39 1010 N/m2, and with a mass density ρ = 2331 kg/m3. The acoustic wave velocity in water has been taken equal to cliq = 1490 m/s. From the optical point of view silicon behaves as an isotropic medium with a refractive index n = 3.5. The surrounding air is considered as a vacuum with a refractive index n = 1 and the refractive index for the water is 1.33. The transmission curves are presented as a function of a normalized frequency Ω = ωa/2πc, where ω is the angular frequency (in s-1), and c is either the velocity of light in vacuum for the electromagnetic waves or the transverse velocity of sound in the direction [001] in silicon for the elastic waves. Phononic crystals In phononic, figure 2(a) represents the transmission coefficient through the water filled structure (black solid lines) together with the transmission through the perfect crystal (red dashed lines) for comparison. The introduction of the water holes leads to new well-defined features at the following reduced frequencies ΩA = 0.301, ΩB = 0.482, and ΩC = 0.627 which appear either as peaks or dips in the transmission coefficient depending on whether they fall in the band gap or in the pass band of the perfect crystal. The calculations of the displacement field at these specific frequencies are reported figure 2(b). One can see clearly that the elastic field is strongly localized inside the water hole meaning that the peaks and dips A, B, and C correspond to eigenmodes of the water hole in interaction with the incident wave. Due to the large contrast between the acoustic velocities and impedances of water and Si, these modes can be considered as cavity resonances inside the holes 2 surrounded by an almost rigid material and, therefore, their frequencies are very close to the solution of the equation J m' (ω r2 / cliq ) = 0 where J’m is the derivative of the Bessel function of order m. (c) PiiilidiMANINNIMMUNNEZ31616111616101iMa; - "71 Vgatilr : Imaiwwwwwwwwww Nadu Ge tano y toal2no-a Reduced frequency (ωa/2πct) The frequencies of the defect modes can be tuned by varying the radius of the water hole. As seen in figure 2(c), when r2/a increases, the frequencies of the defect modes decrease and become closer to each other. Therefore, for the purpose of sensing acoustic velocities, it would be more suitable to do not increase too much the radius of the defect cylinders inside the phononic crystal. 0.6 D C B 0.4 A 0.8 0.2 0.0 0.15 0.20 0.25 0.30 0.35 0.40 0.45 Reduced radius(r2/a) Figure 2: (a) Transmission curve when the radius of the centered cylinder is r2/a=0.25 and filled with water (solid black line). The red dashed curve corresponds to the transmission through a perfect crystal. (b) Maps of the displacement field at the dips A, B and C. (c) Evolution of the frequencies as a function of the radius of the water hole, r2/a. Photonic crystals In photonic we have considered the transverse electric (TE) polarization of the electromagnetic field, i.e. when the electric field is parallel to the plane of periodicity of the crystal (O, x, y). We first calculate the photonic transmission through the structure of figure 1, where the central hole of radius r1/a = r2/a = 0.25 is filled with water. Unfortunately, the introduction of such a defect does not give rise to any peak in the forbidden photonic band gaps. It then appears necessary to change the radius of the defect hole. Figures 3(a) show the (TE) transmission magnified in the reduced frequency range [0.385, 0.435] and through the 2D photonic crystal when the radius of the water hole is taken to be r2/a = 0.4. The transmission spectrum now presents new peaks inside the band gap. The peak (α) at the reduced frequency 0.402 appears with a good quality factor and almost isolated in the middle of the band gap. Figure 3: (a) Photonic transmission for TE polarizations through the photonic crystal for the radius r2/a = 0.4 of the water hole. (b) Map of the electromagnetic fields for the frequencies of the mode α. (c) Evolution of the resonant frequencies inside the photonic band gap as a function of the radius r2/a for TE polarization. The map of its electromagnetic field is presented figure 3(b). In contrast to the phononic modes, the electromagnetic field is not only confined in the water hole but extend in the vicinity of the defect. The evolution of the frequency of (α) peak has been investigated as a function of the radius of the water hole and is presented figure 3(c). One can see the existence of four modes which frequencies increase with the radius of the water hole in good agreement with the decrease of the effective refractive index. Among all these modes only the mode α is well isolated from the others and cover the whole band gap width when varying the radius r2. An equivalent analysis has been performed with the transverse magnetic 3 (TM) polarization of the electromagnetic field, i.e. when the magnetic field is parallel to the plane of periodicity of the crystal (O, x, y), and show the existence of a high quality peak β due to the water hole defect through the (TM) band gap. PhoXonic crystal sensor To make a phoXonic sensor, one needs to design a phononic/photonic structure in which the transmission coefficient displays well-defined features that are very sensitive to the acoustic/optic velocity of the infiltrating liquid. A preliminary condition is that these features should be relatively isolated in order to allow the sensing of the probed parameter on a sufficiently broad range of frequencies. In phononic, figure 2(c) shows that the first two modes A and B satisfy these requirements whatever the radius of the water hole, even though a small radius of the water hole is more favorable. In photonic, as seen in figure 3(c), the radius should be either lower than r2/a = 0.2 or higher than r2/a = 0.35. We will make the demonstration of the dual phononic and photonic sensor dealing with the (α) mode and the radius r2/a = 0.4. The efficiency of the phoXonic sensor has been tested by changing the physical parameters of the liquid filling the central hole. We have considered a set of mixtures of water and 1-propanol at different molar ratio x, for which the density and speed of sound are different, as well as the refractive index, as reported in table 1. Table 1: Density ρliq, speed of sound cliq [10] and refractive index nliq [13] of 1-propanol in water for different molar ratio x. Molar ratio, x Density ρliq (kg/m3) 0 (water) 0.021 0.054 0.056 0.2167 0.230 0.347 0.596 0.6873 998 990 / 974 / 908 881 841 / Longitudinal sound velocity (cliq) (m/s) 1490 1545 / 1588 / 1421 1367 1298 / Refractive index (nliq) 1.333 / 1.3476 / 1.3680 / / / 1.3835 Figure 4(a) and (b) give respectively the evolution of the phononic and photonic (TE) transmission as a function of the liquid filling the central hole (r2/a = 0.4). The transmitted curves are presented in the relevant ranges of frequency which correspond to the presence of the dips A and B in phononic and the peak α in photonic. For both elastic and optical waves, all features associated to the defect modes present a shift in frequency by changing the nature of the liquid. In phononic, the reduced frequencies of the dips A and B increase as soon as the longitudinal velocity of the liquid increases. In photonic, the frequency of the peak α decreases as a function of the refractive index. It means that if we refer to the velocity of sound and light, an increase of the acoustic (optical) velocity leads to a blue shift of the eigenfrequency of the defect. Phononic (a) (b) 0.20 Photonic,TE α -10 Transmission Transmission (dB) 0 -20 -30 1588 m/s 1545 m/s 1490 m/s 1421 m/s 1367 m/s 1298 m/s -40 Α -50 Β -60 0.15 0.18 0.21 0.24 0.27 0.30 0.15 nliq=1.333 nliq=1.3476 0.10 nliq=1.368 nliq=1.3835 0.05 0.00 0.33 0.4020 0.4025 0.4030 (ωa/2πct) (a/λ) Figure 4: (a) Evolution of the phononic transmission curve (in dB) as a function of the acoustic velocity of the liquid filling the central hole of figure 1(a) for the two modes A and B. (b) Evolution of the photonic (TE) transmission curve as a function of the solvent filling the central hole for the (α) mode. 4 The most common way to estimate the efficiency of a sensor is to calculate the sensitivity (S) and the figure of merit (FOM) given, in photonic, by the following expressions: j = S photonic Δλ j Δnliq j FOM photonic = (nm/RIU) S j ×Qj λj (RIU-1) where RIU is the Refractive Index Unit, and Q j is the photonic quality factor of mode j. In phononic the following comparative expressions can be defined: S iphononic = Δfi (MHz/ms-1) Δcliq FOM iphononic = Si × Qi (m/s)-1 fi where Qi is the phononic quality factor of mode i. If we choose to work in the telecommunication range (1550nm), the period of the crystal should be taken equal to 640 nm. With this lattice parameter, and when the central hole is filled with water, the wavelength of modes α occurs at 1590 nm while the frequencies of modes A and B are at 1.78 GHz and 2.74 GHz. Table 2 reports the numerical values of the quality factors, sensitivities and figures of merit for the acoustic and optical modes ((TE) and (TM)) when considering a large (r2 = 0.4a = 256 nm) and a small (r2 = 0.11a = 70 nm) radius of the water filled central hole. The higher sensitivity and figure of merit in phononic are obtained for the smaller radius while in photonic the larger radius leads to the higher sensitivity and figure of merit. Table 2: Phononic and photonic sensitivities (S) and figures of merit (FOM) r2/a (a=640nm) SA 0.40 1.32 0.14 1.73 0.27 58 150 18 175 0.11 3.60 2.25 / / 14 40 6 110 A FOM phononic (m/s) SB -1 B FOM phononic S TE (m/s) -1 FOM TE photonic S TM (RIU-1) FOM TM photonic (RIU-1) 3. OUT-OF-PLANE PROPAGATION Figure 5 represents the 3D schematic view and the elementary unit cell of the investigated structure constituted by a periodically perforated silicon plate arranged in a square array of lattice parameter a in which we consider the excitation of an incident wave normally to the plate. This problem meets the recent interest in the literature about the extraordinary transmission and shielding through sub-wavelength apertures [14-17]. All cylinders present a radius r/a = 0.15 and a thickness of the plate of h/a = 0.25. The plate is embedded in the fluid we want to probe both with acoustic and optical waves. Figure 5: Schematic representation of a normal transmission through a periodically plate embedded in a liquid and the corresponding elementary unit cell used for the calculation. The geometrical parameters are fixed to h/a = 0.25 and r/a = 0.15. PM1s, PBC h 5 Phononic transmission Figure 6(a) represents the acoustic transmission coefficient through the plate when the surrounding medium is filled with water. The transmission curve leads to a well defined peak and dip labeled (A) in figure 6(a). The calculation of the displacement field at the frequency of the dip is represented figure 6(b). An analysis of the component uY and uZ of the displacement field shows that the mode is strongly localized at the circular edge of the water cylinder (see uY(y, z) and uZ(y, z)) with an evanescent decreases outside (see uY(x, y)) and inside the water cylinder (see uZ(x, y)). A schematic representation of the motion of the water cylinder is represented figure 6(c) with regard to the two planes (x, y) and (y, z). The excitation of the eigenmode A leads either to a peak or a dip in the transmission coefficient depending whether the incident wave is in-phase or out-of-phase with the eigenmode oscillation. Therefore, since the peak A followed by a dip is sufficiently well-defined and isolated, it can be seen as a good candidate for the purpose of sensing acoustic velocities. A 0Ai 0.00 0.05 0.10 0.15 015 013 0.30 Red Uü4 05OUenry(ue21q) X Figure 6: (a) Phononic transmission curve for an incidence normal to the plate when the embedded medium is water with the geometrical parameters h/a = 0.25 and r/a = 0.15. (b) Displacement field calculation at the frequency of the dip A. (c) Schematic representation of mode A with the motion of the circular edges of the water cylinder. Photonic transmission In photonic, the calculation of the transmission curve for an incident wave launched perpendicularly to the plate leads also to a well-defined and isolated dip and peak B (figure 7(a)). The map of the electromagnetic field at the frequency of the dip is presented figure 7(b). In contrast with the phononic mode, the electromagnetic field is not only confined inside the water hole but is spread over the whole plate, both inside the water hole and the silicon matrix. One can also notice that the mode is symmetric with respect to the middle plane of the plate. Again, due to its high quality factor and wellisolated frequency, the peak and dip B is suitable for the photonic sensing purpose. Ill Figure 7: (a) Photonic transmission curve for an incidence normal to the plate when the embedded medium is water with the geometrical parameters h/a = 0.25 and r/a = 0.15a. (b) Map of the electromagnetic field at the frequency of the dip B. ri m 6... 6 PhoXonic sensor In this section, we consider the normal transmission through the plate when changing the surrounding fluid medium. We present figure 8(a) and (b) respectively the phononic and photonic transmission spectra for different values of the acoustic velocity and refractive index of refraction of the liquid. For both elastic and optical waves, the two peaks/dips A (phononic) and B (photonic) present a shift in frequency by changing the nature of the liquid. In phononic, the reduced frequency of dip A increases with the longitudinal velocity of the liquid. In photonic, the frequency of dip B decreases as a function of the refractive index. As previously, if we refer to the velocity of sound and light, an increase of the acoustic (optical) velocity leads to a blue shift of the eigenfrequency. If we refer to the same lattice parameter as before (a = 640 nm), the acoustic sensitivity is in the same order of magnitude while in photonic the sensitivity is S = 150nm/RIU, i.e. almost three times higher. (a) (b) 1.0 0.8 Transmission Transmission 0.8 1.0 0.6 0.4 cl =1588 m/s cl =1545 m/s cl =1490 m/s cl =1421 m/s cl =1367 m/s cl =1298 m/s 0.2 0.18 0.6 0.4 0.2 0.0 0.16 water n=1.333 1-butanol n=1.397 13-propanol n=1.438 benzene n=1.5 CS2 n=1.63 0.20 0.22 0.24 0.26 0.28 0.0 0.38 0.30 Reduced frequency(ωa/2πct) 0.39 0.40 0.41 0.42 0.43 0.44 Reduced frequency (a/λ) Figure 8. Representation of the phononic (a) and photonic (b) transmission spectra for respectively different acoustic velocity and refractive index of the liquid. 4. CONCLUSIONS In this work, we investigated the potentiality of an infinite 2D cavity-type phoxonic crystal and a phoxonic plate for sensing the acoustic velocity of a liquid. For sensing applications, the objective has been to design structures in which the phononic/photonic transmission spectra contain well-defined, sufficiently isolated, features, with a high sensitivity to the acoustic/optic parameters of the infiltrating liquid. We studied the first structure for in-plane incident waves and the second when the incoming wave is perpendicular to the phoxonic plate. We identified the physical origin of the specific features appearing in the transmission spectra and discussed the structural parameters useful for making a phoxonic sensor. While the first device is useful for small amount of liquid, in the second one, the plate is totally immersed in the probing fluid, leading to different potential applications. Acknowledgments: This work was supported by the European Commission Seventh Framework Programs (FP7) under the FET-Open project TAILPHOX N° 233883. 7 5. REFERENCES [1] Yablonovitch, E., "Photonic band-gap structures," J. Opt. Soc. Am. 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https://openalex.org/W2306469682	OpenAlex	Open Science	CC-By	2,016	Impact of Timing and Dosage of a Fluoroquinolone Treatment on the Microbiological, Pathological, and Clinical Outcomes of Calves Challenged with Mannheimia haemolytica	Guillaume Lhermie	English	Spoken	11,680	21,160	Impact of timing and dosage of a fluoroquinolone treatment on the microbiological, pathological, and clinical outcomes of calves challenged with Mannheimia haemolytica Guillaume Lhermie, Aude Ferran, Sébastien Assié, Hervé Cassard, Farid El Garch, Marc Schneider, Frédérique Woerhlé, Diane Pacalin, Maxence Delverdier, Alain Bousquet-mélou, et al. To cite this version: Guillaume Lhermie, Aude Ferran, Sébastien Assié, Hervé Cassard, Farid El Garch, et al.. Impact of timing and dosage of a fluoroquinolone treatment on the microbiological, pathological, and clinical outcomes of calves challenged with Mannheimia haemolytica. Frontiers in Microbiology, 2016, 7, pp.1-13. 10.3389/fmicb.2016.00237. hal-02633372 Impact of timing and dosage of a fluoroquinolone treatment on the microbiological, pathological, and clinical outcomes of calves challenged with Mannheimia haemolytica Edited by: Patrick Rik Butaye, Ghent University, Belgium Guillaume Lhermie 1, 2, 3, Aude A. Ferran 2, 4, Sébastien Assié 3, 5, Hervé Cassard 2, Farid El Garch 1, Marc Schneider 1, Frédérique Woerhlé 1, Diane Pacalin 2 †, Maxence Delverdier 2, 6, Alain Bousquet-Mélou 2, 4 and Gilles Meyer 2, 6* Guillaume Lhermie 1, 2, 3, Aude A. Ferran 2, 4, Sébastien Assié 3, 5, Hervé Cassard 2, Farid El Garch 1, Marc Schneider 1, Frédérique Woerhlé 1, Diane Pacalin 2 †, Maxence Delverdier 2, 6, Alain Bousquet-Mélou 2, 4 and Gilles Meyer 2, 6* Reviewed by: Lilia Macovei, The Forsyth Institute, USA Piet Deprez, Ghent University, Belgium 1 Vetoquinol Global Drug Development, Lure, France, 2 Ecole Nationale Vétérinaire de Toulouse, Institut National Polytechnique de Toulouse, Université de Toulouse, Toulouse, France, 3 LUNAM Université, Oniris, UMR BioEpAR, Nantes, France, 4 Institut National de la Recherche Agronomique, UMR1331 ToxAlim, Toulouse, France, 5 Institut National de la Recherche Agronomique, UMR1300 BioEpAR, Nantes, France, 6 Institut National de la Recherche Agronomique, UMR1225 IHAP, Toulouse, France Correspondence: Gilles Meyer g.meyer@envt.fr The efﬁcacy of an early and low inoculum-adjusted marboﬂoxacin treatment was evaluated on microbiological and clinical outcomes in calves infected with 4.107 CFU of Mannheimia haemolytica A1. Twenty-two calves were included based on their rectal temperature rise in the 10 h after challenge and allocated in four groups, receiving a single intramuscular injection of saline (CON), 2 mg/kg marboﬂoxacin 2–4 h after inclusion (early treatment, E2), 2 or 10 mg/kg marboﬂoxacin 35–39 h after inclusion (late treatments, L2, L10). In CON calves, M. haemolytica DNA loads in bronchoalveolar lavages continuously increased from inclusion to day 4, and were associated with persistent respiratory clinical signs and lung lesions. At times of early and late treatments, M. haemolytica loads ranged within 3.5–4 and 5.5–6 log10 DNA copies/mL, respectively. Early 2 mg/kg marboﬂoxacin treatment led to rapid and total elimination of bacteria in all calves. The late treatments induced a reduction of bacterial loads, but 3 of 6 L2 and 1 of 6 L10 calves were still positive for M. haemolytica at day 4. Except for CON calves, all animals exhibited clinical improvement within 24 h after treatment. However, early 2 mg/kg treatment was more efﬁcacious to prevent pulmonary lesions, as indicated by the reduction of the extension and severity of gross lesions and by the histopathological scores. HAL Id: hal-02633372 https://hal.inrae.fr/hal-02633372v1 Submitted on 27 May 2020 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. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. ORIGINAL RESEARCH published: 02 March 2016 doi: 10.3389/fmicb.2016.00237 These results demonstrated for the ﬁrst time that a reduced antibiotic regimen given at an early stage of the disease and targeting a low bacterial load could be efﬁcacious in a natural bovine model of pneumonia. †Present Address: Diane Pacalin, Clinique vétérinaire Le Pont Vert, Mauriac, France †Present Address: Diane Pacalin, Clinique vétérinaire Le Pont Vert, Mauriac, France Specialty section: This article was submitted to Antimicrobials, Resistance and Chemotherapy, a section of the journal Frontiers in Microbiology Specialty section: This article was submitted to Antimicrobials, Resistance and Chemotherapy, a section of the journal Frontiers in Microbiology Received: 25 August 2015 Accepted: 15 February 2016 Published: 02 March 2016 Received: 25 August 2015 Accepted: 15 February 2016 Published: 02 March 2016 Keywords: early treatment, ﬂuoroquinolone, decreased regimen, bovine model, Pasteurellaceae INTRODUCTION et al., 2004; Bywater, 2004). Therefore, a wide consensus in both human and veterinary medicine encourages the reduction of antimicrobial usage to reduce its potential impact on public health. In parallel recent studies have stressed the interests of early therapeutic interventions, when the infectious bacterial load is low, suggesting that the dose of antibiotics may be modulated concurrently with the infectious bacterial load in both human and veterinary medicine (Ferran et al., 2007, 2011; Martinez et al., 2012; Vasseur et al., 2014). For example, in an experimental mouse-lung infection model with Pasteurella multocida, a lower dosage of ﬂuoroquinolone was required to achieve similar bacteriological outcomes when administered “early” compared with a late administration pattern (Ferran et al., 2011). Further, an in vitro study observed high bactericidal activity of marboﬂoxacin even at low concentrations on low inoculum of both M. haemolytica and P. multocida (Illambas et al., 2013). Up to now, such observations have not been reported in a natural host model. These ﬁndings open new perspectives regarding the rational use of antibiotics in farm animals, combining the objectives of reducing their consumption and preserving animal health. In this context, this study aimed to determine the impact of an early and decreased regimen of marboﬂoxacin on the bacteriological burden, the extent of pulmonary lesions and the clinical recovery in calves experimentally infected with M. haemolytica A1. Bovine respiratory disease (BRD) is a major economic concern in young beef cattle production, with a huge economic impact on the farming industry due to production losses and disease treatment and prevention (United State Department of Agriculture, 2013). BRD is mostly multifactorial due to complex interactions between pathogens (virus and/or bacteria and/or fungi), animal (host) factors and non-infectious environmental variables, such as farm management determinants (Fulton, 2009). Among the Gram-negative bacteria, the virulence factors expressed by Mannheimia haemolytica, such as leukotoxin, make it the major bacterial agent of BRD, resulting in high mortality, and loss of productivity in young calves (Van Donkersgoed et al., 1993; Zecchinon et al., 2005). At the herd level, the disease prevalence in suckler calves or young bull production is estimated to be ∼15%, with high variability within herds (Cusack et al., 2003; Assie et al., 2009; Pardon et al., 2011). Recently, M. haemolytica was detected by real-time PCR in France in ∼20% of respiratory samples from BRD (Pelletier, 2013). Bacteria and Culture M. haemolytica A1 strain 11-01-840 was isolated from the lungs of a calf with clinical signs of BRD in 2010 and stored at −80◦C. Prior to each use, one aliquot of the strain was defrosted, and cultured on a Mueller Hinton Agar plate. The MIC was determined in Mueller Hinton Broth in triplicate according to the CLSI reference methods (CLSI, 2009). The MIC of marboﬂoxacin was 0.03 µg/mL. M. haemolytica A1 strain 11-01-840 inoculum was prepared as follows: at day-1 (D-1) prior to inoculation, the strain was incubated overnight in 500 mL of brain heart infusion broth (BHIB) at 37◦C; at day 0 (D0) just prior to inoculation, 50 mL of BHIB was centrifuged at 3000 g for 20 min; pellets were collected in 10 mL of 0.9% NaCl and washed twice in the same buﬀer; and the ﬁnal pellet was then diluted into pre-warmed 0.9% NaCl to obtain a ﬁnal suspension containing 2.105 CFU/mL. Each calf was inoculated with 200 mL of the ﬁnal suspension. Citation: Lhermie G, Ferran AA, Assié S, Cassard H, El Garch F, Schneider M, Woerhlé F, Pacalin D, Delverdier M, Bousquet-Mélou A and Meyer G (2016) Impact of Timing and Dosage of a Fluoroquinolone Treatment on the Microbiological, Pathological, and Clinical Outcomes of Calves Challenged with Mannheimia haemolytica. Front. Microbiol. 7:237. doi: 10.3389/fmicb.2016.00237 March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. INTRODUCTION y Because of the high prevalence of Gram-negative bacteria as involved pathogens, the frequent association between bacteria and viruses, and the absence of pathogen identiﬁcation in ﬁeld conditions, the control of BRD generally includes the use of Gram-negative spectrum antibiotics (Apley, 1997; Fulton, 2009). Two therapeutic strategies are commonly implemented by veterinarians and farmers. Metaphylaxis represents a current practice of early BRD treatment, that consists in treating the entire cohort of animals, in which only a few express clinical signs of BRD (Nickell and White, 2010; Forbes et al., 2011). The advantages of this approach are good survival rate in the cohort and ease of use (Nickell and White, 2010); it also has the disadvantage of exposing to antibiotics a potentially large proportion of animals intended for human consumption even when not required. A second therapeutic approach consists in treating only the aﬀected animals, regardless of the severity of their clinical signs (O’Connor et al., 2013; Heins et al., 2014). This approach limits exposure to antibiotics and decreases the risk of developing resistant strains. However, it is usually seen as the “late approach” because the clinical signs and possibly extensive pulmonary damage have had time to develop prior to treatment, as demonstrated for Pasteurellaceae (Ackermann and Brogden, 2000; Dagleish et al., 2010). Fluoroquinolones, such as marboﬂoxacin, represent a frequent “late approach” treatment for BRD in Europe. Considering the uncertainty regarding the causal agent and time of disease onset, labeled doses are determined by pharmacokinetics (PK) and pharmacodynamics (PD) and dose determination experiments. These doses generally target a cluster of pathogens and consider the highest minimal inhibiting concentration (MIC) reported (Toutain et al., 2002; Valle et al., 2012). These conditions also lead to routine use of a “high” antibiotic dose. Frontiers in Microbiology \| www.frontiersin.org Clinical Examination and Respiratory Sampling Clinical observations were recorded twice a day from 3 days prior to the end of experimentation, between 110 and 134 h (T3) post- challenge. Rectal temperatures were measured every 3 h for a period of 10 h after challenge, then twice daily, and at times of inclusion (T0) and early (T1), and late (T2) treatments. Calves were monitored using a scoring system adapted from Dowling et al. (2002). Scores were assigned as follow: demeanor from 0 to 3 (normal, dull, depressed, or recumbent); rectal temperature from 0 to 2 (<39.5◦C, 39.5–40.5◦C, >40.5◦C); respiratory rate from 0 to 3 (<45, 45–60, 60–90, >90); heart rate from 0 to 2 (<80, 80–100, >100); nasal discharge from 0 to 3 (absent, mild, moderate, or profuse); intensity and added lung sounds from 0 to 2 (normal, increased eﬀort, or labored); and appetite from 0 to 2 (normal, decreased, or anorexic). The clinical score was calculated daily for each calf by adding the values recorded for each parameter. In addition, individual accumulated clinical scores (ACS) were calculated for each calf as the area under daily clinical scores using the trapezoid method. Challenge was performed in calves between 3.5 and 5.5 weeks of age by intratracheal route as previously described (Riﬀault et al., 2010). Brieﬂy, calves were restrained (standing up); an area of 20 cm2 was shaved at the median part of the neckline. Then, a 18 G catheter (Intraﬂon G18, Vycon) was inserted between two tracheal rings ∼10 cm below the larynx. After location veriﬁcation, 200 ml of pre-warmed solution containing 2.105 CFU/mL M. haemolytica was injected into the catheter, corresponding to a total of 4.107 CFU per calf. Calves were allocated into four groups based on elevation of rectal temperatures after challenge. The basal temperature of each calf was calculated as the mean rectal temperature collected individually twice daily for the 3 days prior to challenge. After challenge, rectal temperatures were taken every 3 h for a period of 10 h. Bronchoalveolar lavages (BAL) were performed between 12 and 6 h prior to challenge for all 32 calves and at the end of the experiment for the 22 included calves. In addition, for three calves of each group (CON, E2, L2, and L10), BALs were performed just prior to T1 and T2, the times of early and late treatments, respectively. Experimental Infection of Calves with M. haemolytica Animal experimentation was performed as prescribed by the guidelines of the European Community Council on Animal Care (2010/63/EU), in accordance with accepted human standards of animal care, under ethical agreement number TOXCOM/0021/AF GL (French Ministry of Agriculture, Ethics Committee -C2EA-86). Antimicrobial usage leads to unavoidable selection of resistance in pathogens and commensal bacteria (Courvalin, 2005, 2008). Previous evidence has demonstrated the transfer of resistant bacteria or genetic determinants of resistance between animals and humans (Aarestrup and Wegener, 1999; Angulo Thirty-two dairy Holstein and Normand calves were selected at birth and reared until to 2–3 weeks of age (weight ranged from 47 to 75 kg) in French experimental units with no March 2016 \| Volume 7 \| Article 237 2 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. history of BRD (INRA, Domaine de Borculo, Exmes, France). Calves were colostrum-deprived after birth and fed with an oral rehydration solution the ﬁrst 2 days to avoid transfer of immunoglobulins against bovine respiratory syncytial virus (BRSV), bovine parainﬂuenza virus type (BPI3) as well as M. haemolytica. Then they were fed until day 7 of age with milk replacer and a colostrum substitute (negative for BRSV/BPI3 antibodies), as previously described (Riﬀault et al., 2010). Calves were transported at 2–3 weeks of age and allocated to speciﬁc experimental units in collective pens of three to four animals with free access to hay and fresh water (experimental units of Toulouse, France). They were fed ad libitum with starter food (Passio Floc Junior, SudOuestAliments, Anan, France) and once a day with a milk replacer (Laitine Tech R⃝, Bonilait-protéines, Chasseuneuil du Poitou, France). Upon arrival at the experimental unit, calves received a single intramuscularly injection of E-vitamin and selenium (0.025 mg/kg, Selephérol R⃝, Vetoquinol, France) and underwent a complete physical examination. They remained fully healthy during the 8-day period prior to challenge. Absence of bovine viral diarrhea virus (BVDV) in calves was assessed at birth by negative real-time RT-PCR (Taqvet-BVDV kit, Lifetechnologie). Absence of BRSV and BPI3 was subsequently conﬁrmed by negative serology of calves (LsiVet bovine RSV serum, Life technologies) and negative quantitative RT-PCR (Taqvet BRSV-bPI3 kit, Life technologies) on bronchoalveolar lavages (BAL) at D0 just prior to challenge with M. haemolytica. Absence of M. haemolytica asymptomatic carriers was conﬁrmed by negative qPCR (TaqVet™Triplex P. multocida and M. haemolytica, Life technologies, USA) in nasal swabs 2 days prior to inoculation. Experimental Infection of Calves with M. haemolytica To summarize, E2 calves (early treatment) were treated between 0 and 4 h and L2 and L10 calves (late treatment) between 35 and 39 h post-inclusion, corresponding to 6–10 and 43–48 h after M. haemolytica inoculation. Ten calves were excluded because they failed to develop hyperthermia (mean +1◦C) in the 10-h period after inoculation of M. haemolytica. A description of the experimental design, chronology of sampling and clinical follow-up is presented in Figure 1. The experiment ended 6 days after challenge, between 110 and 134 h post-challenge (time T3). Calves were euthanized during experimentation for ethical reasons or at T3 under general anesthesia overdose (5 mg/kg ketamine, followed by 15 mg/kg pentobarbital sodium and exsanguination). Frontiers in Microbiology \| www.frontiersin.org Clinical Examination and Respiratory Sampling BALs on living animals were performed under general anesthesia (10 mg/kg ketamine and 10 mg/kg diazepam) by endoscopy (Olympus R⃝, CF-EL) as previously described (Deplanche et al., 2007). For each BAL, a same volume of 120 mL of a sterile isotonic sodium chloride solution was injected into the same location of the right lobe. The collected volume varied between 65 and 80 ml per calf and analyses were carried out on 1 mL for each BAL. Titers were expressed by mL of BAL. BALs after euthanasia were performed by direct extraction and lavages of the lungs as previously described (Riﬀault et al., 2010). The recovered suspension was collected and stored at −80◦C directly or after mixing with TRIzol R⃝ (Life Technologies, Saint Aubin, France) for quantitative M. haemolytica PCR analysis. During this 10-h period, 22 of 32 calves were included at time 0 (T0); their rectal temperature exceeded +1◦C their previously basal temperature. Calves were randomly allocated to one of four groups. The control group (4 CON calves) received no antibiotic treatment; the E2 group (6 E2 calves) received an early single intramuscular injection in the neck of 2 mg/kg marboﬂoxacin (Forcyl R⃝, Vetoquinol, Lure, France) at T1, 0–4 h after inclusion (T0). The L2 and L10 groups (6 L2 and L10 calves per group) were treated later at T2, at 35–39 h post-inclusion T0 (33–35 h later than the early treatment), with a single injection of 2 mg/kg or 10 mg/kg marboﬂoxacin, respectively. Necropsy and Tissue Sampling After euthanasia of the calves, gross lesions were photographed and assessed by visual examination and palpation of each of the Necropsy and Tissue Sampling Necropsy and Tissue Sampling After euthanasia of the calves, gross lesions were photographed and assessed by visual examination and palpation of each of the March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 3 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. FIGURE 1 \| Experimental timeline design of the study. FIGURE 1 \| Experimental timeline design of the study. samples were collected at the same location for each calf, including the right and left cranial, the accessory and the cardiac lobes of the lungs and the tracheobronchial lymphatic node. PCR was conducted with 5 µL of elution buﬀer (50 µL) with the TaqVet™Triplex P. multocida and M. haemolytica kit (Life technologies, USA) according the manufacturer’s instructions in a Light Cycler 480 (Roche). A standard plasmid curve for quantiﬁcation was obtained by successive 10-fold dilutions of a plasmid containing 106 DNA copies of the same M. haemolytica ampliﬁed nucleotide sequence (Life technologies, USA). In addition, two housekeeping genes previously mentioned in the literature (gapdh and rpl19) were ampliﬁed in each sample to normalize the qPCR products using GeNorm software (Biogazelle, qBase+). PCR results were analyzed using Roche Light Cycler 480 software (release 1.5.1) using the standard 2nd derivative and ﬁt points methods for all samples (Roche Diagnostics). Results were similar when the PCR threshold was deﬁned by the software (2nd derivative method) or ﬁxed manually (ﬁt points method) around 100 DNA copies. Results are expressed as the mean of PCR titers for each group and each day, after a logistic transformation, or as the accumulated bacterial shed (ABS), which corresponds to the mean area under individual curves of M. haemolytica detected by qPCR in BALs from T0 to T3 (GraphPad Prism software analysis, La Jolla, California, USA). pulmonary lobes. Moreover, the percentage of cranioventral lung consolidation was calculated as described by Amrine et al. (2014), as cranial and ventral lobes are the most common lesion sites. Post-mortem samples were collected for histopathology and bacteriology, including cranial right and left, accessory, cardiac, and caudal left and right lobes of the lungs, as well as the mediastinal and tracheobronchial lymph nodes. For histopathology, tissue samples were paraﬃn wax-embedded after ﬁxation in 10% neutral formalin, sectioned at 3–5 µm and stained with hemalun and eosin. A pathologist described and scored the severity of histopathology and inﬂammation in each slide as either normal (0), mild to moderate (1), or marked (2). Frontiers in Microbiology \| www.frontiersin.org Necropsy and Tissue Sampling A mild to moderate score of 1 was deﬁned by ﬁbrinous alveolitis with macrophagic inﬁltration, moderate quantities of mucus in the lumen of bronchioles and occasional few, small abbesses. A marked score of 2 was deﬁned by hemorrhagic alveolitis with major leucocyte inﬁltration, alteration of neutrophils, foci of necrosis surrounded by oat cells, interlobular edema and thrombosis, moderate quantities of mucus in the lumen of bronchioles, and leucoﬁbrinous pleuresia. A score of histopathological severity was calculated for each calf as the sum of all tissue slide-scores for that animal. M. haemolytica Quantiﬁcation M. haemolytica quantiﬁcation was performed by bacterial titration on post-mortem samples and quantitative PCR on BAL and post-mortem samples of included calves. Statistical Analysis Statistical analyses were performed using GraphPad Prism software (RRID: rid_000081, La Jolla, USA). For DNA quantiﬁcation statistical analyses were performed on logarithmic-transformed data. A two-way ANOVA with repeated measures was used for DNA quantiﬁcation in BAL and for clinical scores. The two-way ANOVA gave a p-value <0.0001 for interaction between time/group (23.7 and 17.1% of total variation respectively). Therefore, we considered that the study had suﬃcient power for further analyses. As eﬀects of the “day” and “treatment” factors were signiﬁcant among interactions, Bonferroni adjustments for multiplicity were used between contrasts to compare the treatments on each day. A For titration, 1 g of the tissue sample collected from the right cranial lobe of the lung was homogenized in 9 mL of 0.9% NaCl. The homogenates were centrifuged at 3000 g for 10 min. The supernatants were discarded, and the pellets were resuspended in 10 mL of 0.9% NaCl. Ten microliters of successive 10-fold dilutions were then plated in triplicate on MH drug-free agar plates for incubation at 37◦C. For all samples, bacterial counts were recorded after 24 h of incubation, and the lowest level of detection of colony counts was set at 100 CFU/g of tissue. For quantitative PCR, DNA was extracted in elution buﬀer from 200 µL of BAL or 100 mg of tissue samples using a Macherey–Nagel nucleospin tissue genomic DNA kit. Five tissue March 2016 \| Volume 7 \| Article 237 4 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. respectively, with 4 of 4, 0 of 6, 4 of 6, and 1 of 6 positive calves in each respective group. two-way ANOVA without repeated measures was used for DNA quantiﬁcation in respiratory tissues. As the group eﬀect was signiﬁcant (p < 0.001, 32.5% of total variation), Bonferroni adjustments for multiplicity test were used to compare the group eﬀect for each tissue. A one-way ANOVA was used to compare the histopathological score, ABS and ACS. As the eﬀect of the “treatment” factor was signiﬁcant for ABS (p = 0.0059) and ACS (p = 0.0168), a Newman–Keuls test was used to compare the treatment eﬀects between groups. No diﬀerences were observed for histopathological score. In parallel, 4 of 4, 2 of 6, 5 of 6, and 4 of 6 calves in the CON, E2, L2, and L10 groups, respectively, were positive for M. Early Treatment with the Low-Dose Led to Rapid and Total Elimination of M. haemolytica in BAL The mean DNA loads of M. haemolytica in BAL over time as well as the mean accumulated bacterial shedding (ABS) scores are presented in Figure 2 for three calves of each group (Supplementary Material). In the CON group, M. haemolytica DNA loads increased regularly over time from inclusion (T0) to the end of the experiment (T3) in all the three tested calves, from 3.3 to 8.6 log10 DNA copies/mL of BAL, suggesting strong replication of the bacteria in the lower respiratory tract. At T1, just prior to early treatment, bacterial DNA loads were similar between the four groups, at approximately 3 log10 DNA copies/mL of BAL. Diﬀerences were then observed according to treatment. In the E2 group, marboﬂoxacin administration at T1 induced a drastic decrease in the bacterial load until euthanasia. At T2, one of the three E2 calves was negative, and all E2 calves were negative at the end of the experiment (T3). In groups L2 and L10, which were treated 33–35 h later, the bacterial load increased in a similar manner as the control group until T2 of late treatment. Then, the bacterial load decreased in both groups, and this decrease appeared to be higher in the L10 group treated with 10 mg/kg marboﬂoxacin. At T3 of euthanasia, only one L10 calf exhibited positive BAL (5.6 log10 DNA copies/mL), whereas all the three L2 calves were positive, with loads ranging from 3.6 to 3.7 log10 DNA copies/mL. Treatment with Marboﬂoxacin Induce Clinical Improvement within 24 h To determine whether bacterial growth was associated with change of clinical status, we assessed calves for clinical signs and lesions over time. All calves were healthy prior to and immediately after inoculation. At this time, the mean rectal temperature was 38.8 ± 0.4◦C. Twenty-two of thirty-two calves were selected based on rectal temperature elevation. These calves were randomly allocated into one of four groups just after receiving the respective treatment. The mean M. haemolytica DNA loads were signiﬁcantly diﬀerent between the CON group and the E2, L2, L10 groups at T3, between E2 and L2 at T3, and between E2 and L10 at T2 (Figure 2A). In addition, ABS scores in BAL indicated signiﬁcant diﬀerences between the CON and the E2 and L2 groups and between the E2 and L2 groups (Figure 2B). Rectal temperature of the included calves peaked between 3 and 9 h after challenge. Mean rectal temperature peaked at 40.2◦C at the time of inclusion in all groups. Then, rectal temperature decreased over time in all groups. Rectal temperature decreased to baseline within 6 h after early treatment in E2 group and within 12 h after late treatment in the L2 and L10 groups. In the non-treated animals, rectal temperature remained above the temperature of the treated groups at each time point, indicating a signiﬁcant diﬀerence between E2 calves and non-treated calves at T2 (data not shown). Statistical Analysis haemolytica in at least one of the 5 respiratory tissues (right and left cranial lobes, accessory and cardiac lobes, tracheobronchial lymphatic node) tested in each calf. Large diﬀerences were observed when considering the number of positive samples per calf and per group. In the E2 group, only the intermediate lobe of one calf and the left cranial lobe of another calf were positive, with moderate titers of 3.8 and 3.9 log10 DNA copies/100 mg (Figure 3). In contrast, 80% of the tissue samples of the CON group were positive, whereas the L2 and L10 groups similarly had 37% of tissue samples as positive. In the CON group, high DNA individual titers, ranging between 4.6 and 8.2 log10 DNA copies/100 mg, were detected in positive samples (Figure 3). The situation was intermediate in the L2 and L10 groups, with mean loads ranging from 0.7 to 3.2 log10 DNA copies/100 mg, depending on the type of tissue. The mean M. haemolytica DNA loads in each tissue samples (Figure 3) also indicated signiﬁcant diﬀerences between the CON and E2 group, despite large intra- group variability. To conﬁrm the results obtained by qPCR in tissue samples, a direct bacterial culture and titration was performed on the right cranial lobe of the lungs (Table 1). In the E2 group, all samples collected for bacterial culture remained negative, even the two samples that were slightly positive with PCR. For the CON, L2, and L10 groups, the M. haemolytica counts globally mimicked those obtained by PCR, except for two calves in the L2 group for which culture was slightly positive but not with PCR analysis (Table 1). Distribution and Loads of M. haemolytica in Respiratory Tissues Showed Differences between Marbofoxacin Treatments To further investigate the PCR results, we tested all the 22 included calves for the presence of M. haemolytica in BAL and in post-mortem tissue samples at T3 of euthanasia (Supplementary Material). The mean DNA loads in BAL were 7, 0, 2.3, and 1 log10 DNA copies/mL for calves in the CON, E2, L2, and L10 groups, Clinical signs started at 6 ± 3 h and peaked between 24 and 40 h after inoculation for most calves. After peak, clinical signs of CON calves slightly decreased throughout the course of infection. In the other groups, clinical signs were very similar to those observed in the CON group until March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 5 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. Lhermie et al. Optimization of An TABLE 1 \| Main individual results of clinical and bacteriological examination after M. haemolytica challenge for untreated calves (CON) or calves treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion. Treatment group Calf Cranio ventral percentage lung consolidation % of lung lobes with microscopic lesions Highest individual clinical score Accumulated clinical score M. haemolytica in right cranial lobe (log10 CFU/g) M. haemolytica in right cranial lobe (log10 DNA copies/g) M. March 2016 \| Volume 7 \| Article 237 Distribution and Loads of M. haemolytica in Respiratory Tissues Showed Differences between Marbofoxacin Treatments haemolytica in BAL at necropsy (log10 DNA copies/mL) C 4254 88 75 10 34.8 10.5 9.2 8.6 C 4269 0 0 3 11.8 3.0 6.0 7.0 C 4310 euthanized at 56 h 51 87.5 10 24.8 10.0 6.7 7.9 C 4312 12 25 6 20 4.0 7.7 5.5 E2 4253 0 0 3 5.5 0.0 0.0 0.0 E2 4261 0 0 6 9.5 0.0 0.0 0.0 E2 4276 0 0 9 11.5 0.0 0.0 0.0 E2 4284 0 0 3 6.7 .0 0.0 0.0 E2 4298 17 25 5 5.7 0.0 0.0 0.0 E2 4321 0 0 5 8.4 Contamination 0.0 0.0 L2 4263 0 0 8 17.5 3.2 0.0 0.0 L2 4280 55 87.5 12 28.7 8.8 9.4 3.7 L2 4295 0 12.5 6 11.2 0.0 5.3 3.7 L2 4311 11 25 5 9.7 3.7 2.0 4.0 L2 4316 0 0 6 8.4 2.7 2.0 1.2 L2 4319 0 25 8 10.1 0.0 0.0 0.0 L10 4275 0 0 6 10.3 3.2 5.1 5.7 L10 4292 4 37.5 9 14.5 Contamination 4.0 0.0 L10 4306 0 0 10 13.9 0.0 0.0 0.0 L10 4307 0 0 5 8.1 0.0 3.5 0.0 L10 4309 3 25 11 19.9 3.0 5.3 4.1 L10 4313 0 12.5 8 11.9 0.0 0.0 0.0 Individual clinical scores were calculated as the sum of followings: demeanor from 0 to 3 (normal, dull, depressed, or recumbent); rectal temperature from 0 to 2 (<39.5◦C, 39.5–40.5◦C, >40.5◦C); respiratory rate from 0-3 (<45, 45–60, 60–90, >90); heart rate from 0 to 2 (<80, 80–100, >100); nasal discharge from 0 to 3 (absent, mild, moderate, or profuse); intensity and added lung sounds from 0 to 2 (normal, increased effort, or labored); and appetite from 0 to 2 (normal, decreased, or anorexic). Accumulated clinical scores (ACS) were calculated as the area under daily clinical scores using the trapezoid method. The percentage of cranioventral lung consolidation was calculated as described by Amrine et al. (2014). M. haemolytica quantiﬁcation was done using qPCR in a Light Cycler 480 (Roche). A standard curve for quantiﬁcation was obtained by successive 10-fold dilutions of a plasmid containing the same M. haemolytica ampliﬁed nucleotide sequence. ge for untreated calves (CON) or calves treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after March 2016 \| Volume 7 \| Article 237 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. Distribution and Loads of M. haemolytica in Respiratory Tissues Showed Differences between Marbofoxacin Treatments FIGURE 2 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in bronchoalveolar lavages (A) and mean accumulated bacterial shed (B) in calves treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion. The black and red arrows indicate the time of early and late treatments respectively. Signiﬁcant differences: p < 0.05; p < 0.01; p < 0.001. FIGURE 2 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in bronchoalveolar lavages (A) and mean accumulated bacterial shed (B) in calves treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion. The black and red arrows indicate the time of early and late treatments respectively. Signiﬁcant differences: p < 0.05; p < 0.01; p < 0.001. FIGURE 3 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in respiratory post-mortem tissues of calves not treated (■), treated with 2 mg/kg marboﬂoxacin at 4 h after inclusion (E2, ), or 2 mg/kg (L2, ), or 10 mg/kg (L10, ) marboﬂoxacin at 36 h after inclusion. Signiﬁcant differences are shown in the table: p < 0.05; *p < 0.01. FIGURE 2 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in bronchoalveolar lavages (A) and mean accumulated bacterial shed (B) in calves treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion. The black and red arrows indicate the time of early and late treatments respectively. Signiﬁcant differences: p < 0.05; p < 0.01; p < 0.001. FIGURE 3 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. Frontiers in Microbiology \| www.frontiersin.org DISCUSSION The objective of this study was to assess the eﬃcacy of a decreased marboﬂoxacin regimen when administered at early-time of infection, on bacterial burden and clinical outcomes in calves infected with M. haemolytica. To fulﬁll this objective, we ﬁrst developed an experimental model for the detection and treatment of M. haemolytica mimicking ﬁeld conditions. BRD treatments are frequently administered after visual appraisal of clinical respiratory signs. However, because these signs often occur late in the disease course (Schaefer et al., 2007), hyperthermia is increasingly frequently used to promptly detect and treat BRD in young cattle. In a French ﬁeld study conducted in young bulls, Timsit et al. (2011) demonstrated that episodes of hyperthermia (>39.7◦C) preceded the onset of nasal discharge, abnormal lung sounds, and depression by a mean of 19, 38.8, and 51.2 h, respectively. To be consistent with these ﬁeld observations, we delayed early and late treatments by 30 h. Moreover, we included calves for early treatments based on an increase in rectal temperature of 1◦C compared to their basal rectal temperature. This 1◦C implementation matches with the temperature threshold commonly used for treatment and previously reported as an early sign of BRD (Galyean et al., 1995; Schaefer et al., 2007; Timsit et al., 2011). At the same time, a challenge dose of 4 × 107 CFU/calf was used to mimic the clinical signs of a natural infection. First, in a previous pilot study, we tested two doses: 1010 and 107 CFU/calf using the same model of infection (data not shown). The high inoculum lead to a severe superacute disease inducing death in the ﬁrst 24 h, as observed in other experimental models using bacterial load infused in lungs over 109 CFU (Fajt et al., 2004; Burciaga- Robles et al., 2010; Hanzlicek et al., 2010; Rose-Dye et al., 2011). In contrast, the 107 CFU dose was associated with a moderate to severe disease evolving in 4–5 days and allowed longer clinical and bacteriological monitoring at the infectious site. Secondly, the qPCR threshold (Ct) values of non-treated calves of these two studies ranged between 19–32 and 19.9–35 in the BAL T2 or T3 and the lung tissues, respectively. These values ﬁt with those obtained (Ct range of 24–36) by the same method for the ﬁeld BRD diagnostic during a winter season (personal data from 105 samples with 24 samples positive for M. haemolytica). Distribution and Loads of M. haemolytica in Respiratory Tissues Showed Differences between Marbofoxacin Treatments haemolytica DNA copies (with standard deviations) in respiratory post-mortem tissues of calves not treated (■), treated with 2 mg/kg marboﬂoxacin at 4 h after inclusion (E2, ), or 2 mg/kg (L2, ), or 10 mg/kg (L10 ) marboﬂoxacin at 36 h after inclusion Signiﬁcant differences are shown in the table: p < 0 05; *p < 0 01 FIGURE 3 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in respiratory post-mortem tissues of calves not treated (■), treated with 2 mg/kg marboﬂoxacin at 4 h after inclusion (E2, ), or 2 mg/kg (L2, ), or 10 mg/kg (L10, ) marboﬂoxacin at 36 h after inclusion. Signiﬁcant differences are shown in the table: p < 0.05; *p < 0.01. FIGURE 3 \| Mean titers (qPCR, Roche Light Cycler 480, standard 2nd derivative method) of M. haemolytica DNA copies (with standard deviations) in respiratory post-mortem tissues of calves not treated (■), treated with 2 mg/kg marboﬂoxacin at 4 h after inclusion (E2, ), or 2 mg/kg (L2, ), or 10 mg/kg (L10, ) marboﬂoxacin at 36 h after inclusion. Signiﬁcant differences are shown in the table: p < 0.05; *p < 0.01. two CON calves over time and one L2 calf and one L10 calf prior to treatment. In the CON group, one of the two calves with severe disease was euthanized at 56 h after infection for ethical reasons. Moderate signs included hyperthermia, each treatment. Consequently, for clinical score evaluation, all calves were considered as a unique group until marboﬂoxacin administration, as shown in Figure 4A. To summarize, the clinical signs were moderate in most of calves and severe in March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 7 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. tachypnea and dyspnea with labored breathing. In severe forms, reduced appetite, depression, recumbence, and severe dyspnea with labored breathing and abnormal lung sounds (wheezes and crackles) were observed. the L2 and L10 groups. In addition, histopathological lesions of sub-acute bronchopneumonia (score of 1) were also observed but with less frequency in only one calf in each of the four included groups. In this case, lesions were characterized by a ﬁbrinous and macrophagic alveolitis associated with small abscesses or pyogranulomas of the lung parenchyma (Figure 5D). DISCUSSION Finally the choice of the two doses of marboﬂoxacin was based on a PK/PD evaluation of antimicrobial eﬃcacy previously published for M. haemolytica (Valle et al., 2012; Potter et al., 2013). The The accumulated clinical score (ACS) was evaluated for each calf of each group from the time of infection until the time of euthanasia (T3). The mean ACS for each group indicated strong diﬀerences between the treated and non-treated groups, with signiﬁcant diﬀerences only between the E2 and CON groups (Figure 4B). Frontiers in Microbiology \| www.frontiersin.org Distribution and Loads of M. haemolytica in Respiratory Tissues Showed Differences between Marbofoxacin Treatments The mean clinical scores indicated that marboﬂoxacin injections were followed by a rapid (within 24 h) clinical improvement of calves, regardless of early or late treatment (Figure 4A). Twenty-four hours after each respective treatment, 4 of 6 E2 and L2 calves and all of the L10 calves presented a clinical score less than 4 (Table 1). Forty-eight hours after treatment up to the end of the experiment, clinical scores remained <4 for all calves in the treated groups. Signiﬁcant diﬀerences were observed between the non-treated animals and the E2 group at 24 h, between the CON group and E2, L2, and L10 groups at 48 and 72 h after infection, and ﬁnally between the CON group and the E2 and L10 groups at 96 h post infection (Figure 4A). The mean histopathological score for each group is shown in Figure 5E, with strong but no signiﬁcant diﬀerences between the treated and not treated calves, except between the E2 and CON groups. This mean score was correlated with the total cranioventral percentage of lobe consolidation (Figure 5F), except for one E2 calf that exhibited a low histopathological score, despite moderate consolidation. Early Treatment with the Low-Dose Prevent Pulmonary Lesions Lung lesions were primarily observed in all calves in the CON group. Lung damage was observed in the cranial lobes and was typical of acute infection with M. haemolytica, including a high tissue consolidation and a dark-red to gray-red lobular coloration with foci of necrosis. Lung lesions were also frequently associated with pleuresia and ﬁbrin deposits. The mean percentage of cranioventral lung consolidation was 38, 5, 11, and 1% in the CON, E2, L2, and L10 groups, respectively. However, the extension of the gross lesions varied greatly between animals. In the CON group, two calves exhibited severe lesions on both cranial and caudal lobes, one calf exhibited lesions on cranial lobes only, and one calf presented only very restricted lesions. In all groups, calves with the most severe lung lesions also presented the highest individual clinical score and high M. haemolytica content in the lungs (Table 1). At histopathology, 75, 16.7, 66.7, and 50% of CON, E2, L2, and L10 calves, respectively, had microscopic lesions in at least one lobe of the lung. Considering the extension of the microscopic lesions, 46.9, 4.2, 22.9, and 12.5% of the lobes were positive in the CON, E2, L2, and L10 groups, respectively. In the CON group, microscopic lesions were located in all lung lobes for two of four calves and only in cranial lobes of the two other animals. By comparison with normal lung microscopic examination (Figure 5A), lesions were speciﬁc to an acute severe bronchopneumonia and characterized by an hemorrhagic alveolitis with large ﬁbrinous deposits and major inﬁltration of leucocytes (Figure 5B), foci of necrosis surrounded by oat cells and altered neutrophils (Figure 5C), interlobular edema and thrombosis (Figure 5C), moderate quantities of mucus in the lumen of bronchioles and leucoﬁbrinous pleuresia (score of 2). This type of lesion (score of 2) was not observed for E2 calves but was observed to a small extent in three of the six calves in March 2016 \| Volume 7 \| Article 237 8 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. FIGURE 4 \| Mean clinical scores (A) and mean accumulated clinical scores (B) of calves inoculated with M. haemolytica and treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion (black arrow) or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion (red arrow). Early Treatment with the Low-Dose Prevent Pulmonary Lesions haemolytica A1 strain used in this study, which was previously conﬁrmed to be highly replicative and pathogenic in calves. p p p g Compared with the early treatment, calves treated 30 h later with the same 2 mg/kg dose of antibiotic displayed a diﬀerent pattern. Signiﬁcant diﬀerences in DNA titers were found between the E2 and L2 groups in BALs at T3 and in accumulated bacterial loads. Large diﬀerences in M. haemolytica DNA titers, although not signiﬁcant, were also observed between the two groups considering both the number of calves with positive lung samples and the total number of positive tissues per calf. The situation appears to be slightly diﬀerent when calves were treated later but with a high 10 mg/kg dose of marboﬂoxacin. No signiﬁcant diﬀerences were observed between the E2 and L10 groups at T3 of euthanasia. In the L10 group, M. haemolytica was detected in 10 of 30 respiratory samples but in only 2 of 6 tested calves. Further, even if bacterial loads were similar in the L2 and L10 calves just prior to treatment (T2), only one L10 calf presented moderate bacteria DNA titers in BAL 3 days after treatment (T3), whereas all the three L2 calves remained positive, suggesting that an increased marboﬂoxacin dosage from 2 to 10 mg/kg was associated with higher bacterial elimination when treatments area initiated later. This result is also supported by the decrease in mean DNA titers shown in Figure 2, suggesting that clearance of bacteria was faster for the L10 than L2 group and was likely similar between the L10 and E2 groups. These results are in agreement with previous works, conﬁrming that ﬂuoroquinolones display a concentration-dependent killing mechanism of M. haemolytica (Sarasola et al., 2002; Illambas et al., 2013). Finally, our results simultaneously conﬁrm the concentration-dependent behavior of ﬂuoroquinolones against Gram-negative bacteria for a given bacterial load, and suggests that the size of the bacterial load at the time of treatment inﬂuences the range of exposure magnitude required to obtain bacterial eradication. Our model of M. haemolytica infection showed that a single early treatment with a single dose of 2 mg/kg marboﬂoxacin led to the rapid elimination of bacteria in all infected calves between 30 and 116 h after injection. All E2 calves were negative for M. haemolytica detection in BAL at the end of the experiment. Early Treatment with the Low-Dose Prevent Pulmonary Lesions high dose of 10 mg/kg matched with the current recommended dose of marboﬂoxacin, as mentioned in the commercial product (FORCYL R⃝) with the indication for treatment of BRD. The low dose was the lowest dose ensuring bactericidal eﬃcacy in vitro on a targeted low inoculum of M. haemolytica (Lhermie et al., 2015). (Lhermie et al., 2015). These results indicated that marboﬂoxacin was much more eﬃcacious against M. haemolytica than P. multocida, in a mouse model which is not a natural host of these pulmonary pathogens. In the natural host model tested in the present study, which is particularly sensitive to the virulence factors expressed by M. haemolytica such as leukotoxin (Van Donkersgoed et al., 1993; Apley, 1997; Zecchinon et al., 2005), we conﬁrmed the ability of an early marboﬂoxacin treatment to produce rapid and complete microbiological cure. To our knowledge, this is the ﬁrst study assessing the kinetics of M. haemolytica loads in the lower respiratory tract of calves for more than 5 consecutive days. Our results in the control group (Figure 2) indicated a continuous increase in bacterial loads in the BAL from the start to the end of the experiment, conﬁrming our model of infection. Unfortunately it was not possible to strengthen our PCR results by bacterial load counts for practical reasons. Nevertheless, these assessments were performed using a commercialized qPCR kit that is highly repeatable and reproducible and that is increasingly used to diagnose M. haemolytica in the ﬁeld (Pelletier, 2013). PCR or qPCR methods were also shown to detect signiﬁcantly higher number of positive cases than culture for M. haemolytica or H. somni (Tegtmeier et al., 2000; Angen et al., 2009; Bell et al., 2014) and much faster (Guenther et al., 2008). To our knowledge, only another study assessed M. haemolytica growth during the 48 h after infection with a similar inoculum load (108 CFU/calf). The authors observed an increased bacterial count in bronchial secretions until 18 h post-infection, with a plateau of 107 CFU/mL until the end of the experiment (Sarasola et al., 2002). Diﬀerences of bacteria kinetics between the two studies may be due to the method of quantiﬁcation because part of the DNA for PCR titration may have been obtained from bacteria killed by the immune response of calves. Another possibility is virulence diﬀerence of the M. Frontiers in Microbiology \| www.frontiersin.org Early Treatment with the Low-Dose Prevent Pulmonary Lesions For (A), calves remained in the control group until the time of treatment. For (B), individual ACS were determined as the area under the curve of clinical scores collected at −16, 6, 24, 40, 72, and 96 h (Graph Pad software, La Jolla, USA). Signiﬁcant differences: p < 0.05; p < 0.01; p < 0.001. FIGURE 4 \| Mean clinical scores (A) and mean accumulated clinical scores (B) of calves inoculated with M. haemolytica and treated with 2 mg/kg (E2) marboﬂoxacin at 4 h after inclusion (black arrow) or 2 mg/kg (L2) or 10 mg/kg (L10) marboﬂoxacin at 36 h after inclusion (red arrow). For (A), calves remained in the control group until the time of treatment. For (B), individual ACS were determined as the area under the curve of clinical scores collected at −16, 6, 24, 40, 72, and 96 h (Graph Pad software, La Jolla, USA). Signiﬁcant differences: p < 0.05; p < 0.01; *p < 0.001. FIGURE 5 \| Microscopic examination. (A) Normal lung microscopic examination (hemalun and eosin, x200). (B) Lesions of hemorrhagic alveolitis, large ﬁbrinous deposits, and major inﬁltration of leucocytes, score of 2 (hemalun and eosin, ×200). (C) oat cells and altered neutrophils, score of 2 (hemalun and eosin, ×200). (D) Macrophagic alveolitis, score of 1 (hemalun and eosin, ×200). (E) Mean histopathological score (+/−sd) of calves challenged with M. haemolytica and treated with 2 or 10 mg/kg marboﬂoxacin at the early or late stages of infection. (F) Correlation between the total cranioventral percentage of lobe consolidation (gross lesions, × axis) and number of lobes with microscopic lesions (y-axis). FIGURE 5 \| Microscopic examination. (A) Normal lung microscopic examination (hemalun and eosin, x200). (B) Lesions of hemorrhagic alveolitis, large ﬁbrinous deposits, and major inﬁltration of leucocytes, score of 2 (hemalun and eosin, ×200). (C) oat cells and altered neutrophils, score of 2 (hemalun and eosin, ×200). (D) Macrophagic alveolitis, score of 1 (hemalun and eosin, ×200). (E) Mean histopathological score (+/−sd) of calves challenged with M. haemolytica and treated with 2 or 10 mg/kg marboﬂoxacin at the early or late stages of infection. (F) Correlation between the total cranioventral percentage of lobe consolidation (gross lesions, × axis) and number of lobes with microscopic lesions (y-axis). March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 9 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. Early Treatment with the Low-Dose Prevent Pulmonary Lesions Only 2 out of 30 tissues samples contained low DNA quantities (<4 log10 copies/100 mg). The rapid clearance of M. haemolytica after early treatment with a lower marboﬂoxacin dose than that currently recommended (10 mg/kg) might be explained by the relatively low loads of bacteria observed in BALs at T1, suggesting that the required regimen to obtain bacterial elimination is dependent (partly or mostly) on the size of the inoculum at the time of treatment. These results are supported by those obtained in various in vitro and in vivo studies conducted with ﬂuoroquinolones, stressing the inﬂuence of bacterial cell density on the eﬃcacy of these antibiotics (Mizunaga et al., 2005; Ferran et al., 2007; Kesteman et al., 2009; Udekwu et al., 2009). Moreover, in a recent experimental study testing bactericidal eﬃcacy of marboﬂoxacin in a mouse-lung infection model using either M. haemolytica or P. multocida strains with close marboﬂoxacin MICs, we observed that the administration of a lower dose of marboﬂoxacin (given 10 h after inoculation) led to total eradication of a bacterial load of 108 CFU/lung of M. haemolytica. In contrast, a similar dose completely failed to achieve bacterial elimination of a bacterial load of 108 CFU/lung of P. multocida Taken together, these results from an experimental model highlight the beneﬁts of using early treatments with a low dose of antibiotic that should lead to a fast cure of M. haemolytica infection in the lower respiratory tract. One major question for livestock professionals is the impact of such early treatment on animal health and also human health by limiting antibiotic consumption. In this study, we only focused on animal health by assessing clinical signs and lesions of infected and treated calves. In the control group, rectal temperature and mean clinical score peaked within 3–12 and 24–40 h following challenge, respectively, reached a plateau, and then slightly decreased until the end of the experiment. This mimics the precedence of hyperthermia as observed in ﬁeld conditions and supported in numerous respiratory challenge models (Fajt et al., 2004; Burciaga-Robles et al., 2010; Hanzlicek et al., 2010; Forbes et al., 2011; Rose-Dye et al., 2011; Crouch et al., 2012). In our challenge March 2016 \| Volume 7 \| Article 237 10 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. model reproducing moderate to severe disease, early or late treatments with marboﬂoxacin clearly induced recovery of all calves in <24 h. Early Treatment with the Low-Dose Prevent Pulmonary Lesions These results are supported by those obtained in ﬁeld studies that have monitored the eﬀects of marboﬂoxacin on rectal temperature and clinical status of animals naturally infected with various respiratory pathogens (Thomas et al., 2001; Grandemange et al., 2012). Total ACS showed major diﬀerences between the control and E2, L2, and L10 groups; however, these diﬀerences were only signiﬁcant between the E2 and control groups (Figure 4B). The main explanations may be the variability of individual response in the challenge model or more likely, the delay of treatment, as clinical scores decreased similarly in all treated groups within 24 h after antibiotic injection, with signiﬁcant diﬀerences compared with the control group (Figure 4A). decreased antimicrobial regimen could be considered in ﬁeld conditions. We recently performed a ﬁeld study to evaluate antimicrobial consumption and therapeutic eﬃcacy of an early vs. late treatment protocol with a similar decreased antimicrobial regimen (marboﬂoxacin 2 or 10 mg/kg) in 195 young bulls with BRD. Our results suggest that the combination of an early detection of disease (increase of body core temperature) with a decreased marbofoxacin treatment reduces drug consumption at the herd level without aﬀecting treatment eﬃcacy compared to a standard therapy. (Lhermie et al., submitted). Such ﬁndings evidence the possible rationalization of antimicrobial use under ﬁeld conditions, to limit impact on public health. To conclude, to our knowledge, this is the ﬁrst time that a decreased antimicrobial regimen administered at early time of infection and targeting a low bacterial load demonstrated eﬃcacy in a bovine natural host experimental model. The responses to the treatments, in terms of bacterial elimination, pulmonary lesions, and clinical recovery, were similar between the lower dose (low inoculum-adjusted) administered at an early time of infection and the higher dose (high inoculum-adjusted) given at a later time of infection. Altogether, our results are proof of principle that there is a place for the optimization of antibiotic doses in food-producing animals, provided strict conditions of early diagnosis of illness and/or bacterial load that allow combining the cure of bacterial infections with the reduction of antibiotic consumption. Regarding antimicrobial consumption under public health considerations, further studies evaluating the beneﬁts of modulating dose regimens have to be encouraged. Our results also suggest that early treatment may prevent pulmonary lesions. ACKNOWLEDGMENTS We would like to thank C. Foret, A. Teillaud and M. Moulignié for technical assistance and Y. Gallard and S. Ronsin (INRA UE0326 DEP Domaine Expérimental du Pin) for providing and nursing colostrum-deprived calves and D. Concordet for statistical analysis comments. Early Treatment with the Low-Dose Prevent Pulmonary Lesions Whatever the antibiotic dosage, treatment led to a low frequency of lung lesions, but signiﬁcant diﬀerences for histopathological scores were only observed between the E2 and control groups. Further, calves treated earlier presented a lower percentage of cranioventral lung consolidation, a lower number of lobes with macroscopic and microscopic lesions, and lower histopathological scores compared with calves treated 30 h later. Again, these results could be associated with rapid elimination of bacterial burden, avoiding bacterial growth and thus the onset of pulmonary damage. We also observed that lesion severity was correlated with bacterial load in the lungs. Our results agree with those reported in another experimental BRD model, in which a positive relationship between the severity of clinical signs, cumulative clinical scores, and extension of pulmonary macroscopic and microscopic lesions was observed (Forbes et al., 2011). FUNDING This work was supported by the Vetoquinol Company (Lure, France). The funders (Vetoquinol) had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Promoting a dose modulated concurrently with the infectious bacterial load could be diﬃcult to implement in cattle production under actual ﬁeld conditions. Indeed, quantiﬁcation of bacterial content in respiratory samples is not currently performed, although the present qPCR is now used in several veterinary laboratories (Pelletier, 2013). The main caveat is the minimum 1 day-time to obtain a quantitative result, which implies that the treatment would delivered with a delay, when bacterial loads would be already higher than when initially tested. However, constantly more rapid and easier techniques are being developed for detection and quantiﬁcation of bacteria such as LAMP- PCR, biosensors, reﬂectance spectroscopy, raman spectroscopy etc....(for review Noble and Weisberg, 2005; Velusamy et al., 2010). Since our model and another study (Sarasola et al., 2002) showed a correlation between the bacterial content and the apparition and evolution of the disease, on-ﬁeld strategy based on early diagnosis of illness (with eﬃcient markers) and using AUTHOR CONTRIBUTIONS All authors listed, approved the work for publication. GL, AB, and GM designed the study; GL, DP, HC, AF, MD, and GM performed the experiments; GL, AF, SA, FG, MS, FW, and GM analyzed the data; GL, AB, and GM wrote the manuscript. All authors listed, approved the work for publication. GL, AB, and GM designed the study; GL, DP, HC, AF, MD, and GM performed the experiments; GL, AF, SA, FG, MS, FW, and GM analyzed the data; GL, AB, and GM wrote the manuscript. However, it is unclear why, despite increasing high qPCR titers in the lungs, we did not observe severe disease with extensive pulmonary lesions in some of the non-treated animals. This observation suggests that implementation of on-ﬁeld therapy based on early diagnosis of illness and using decreased antimicrobial regimen may require speciﬁc diagnosis tools to measure bacterial load and/or, if possible, a precise monitoring of markers correlated with early infection stage. Frontiers in Microbiology \| www.frontiersin.org REFERENCES A., and Malcolm-Callis, K. J. (1995). Eﬀects of arrival medication with tilmicosin phosphate on health and performance of newly received beef cattle. J. Anim. Sci. 73, 1219–1226. 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SUPPLEMENTARY MATERIAL The Supplementary Material for this article can be found online at: http://journal.frontiersin.org/article/10.3389/fmicb. 2016.00237 March 2016 \| Volume 7 \| Article 237 11 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. REFERENCES A review of technologies for rapid detection of bacteria in recreational waters. J. Water Health 3, 381–392. doi: 10.2166/wh.2005.051 Dowling, A., Hodgson, J. C., Schock, A., Donachie, W., Eckersall, P. D., and McKendrick, I. J. (2002). Experimental induction of pneumonic pasteurellosis March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 12 Optimization of Antibiotic Dosage Regimen in Calves Lhermie et al. O’Connor, A. M., Coetzee, J. F., Da Silva, N., and Wang, C. (2013). A mixed treatment comparison meta-analysis of antibiotic treatments for bovine respiratory disease. Prev. Vet. 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Res. 36, 133–156. doi: 10.1051/vetres:2004065 Schaefer, A. L., Cook, N. J., Church, J. S., Basarab, J., Perry, B., Miller, C., et al. (2007). The use of infrared thermography as an early indicator of bovine respiratory disease complex in calves. Res. Vet. Sci. 83, 376–384. doi: 10.1016/j.rvsc.2007.01.008 Conﬂict of Interest 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. Tegtmeier, C., Angen, O., and Ahrens, P. (2000). Comparison of bacterial cultivation, PCR, in situ hybridization and immunohistochemistry as tools for diagnosis of Haemophilus somnus pneumonia in cattle. Vet. Microbiol. 76, 385–394. doi: 10.1016/S0378-1135(00)00259-5 Copyright © 2016 Lhermie, Ferran, Assié, Cassard, El Garch, Schneider, Woerhlé, Pacalin, Delverdier, Bousquet-Mélou and Meyer. 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. Frontiers in Microbiology \| www.frontiersin.org REFERENCES No use, distribution or reproduction is permitted which does not comply with these terms. Thomas, E., Caldow, G. L., Borell, D., and Davot, J. L. (2001). A ﬁeld comparison of the eﬃcacy and tolerance of marboﬂoxacin in the treatment of bovine respiratory disease. J. Vet. Pharmacol. Ther. 24, 353–358. doi: 10.1046/j.1365- 2885.2001.00333.x Timsit, E., Bareille, N., Seegers, H., Lehebel, A., and Assie, S. (2011). Visually undetected fever episodes in newly received beef bulls at a fattening operation: March 2016 \| Volume 7 \| Article 237 Frontiers in Microbiology \| www.frontiersin.org 13	7,000
W4210806986.txt_5	German-Science-Pile	Open Science	Various open science	null	None	None	Unknown	Unknown	8,415	17,000	Im Gegensatz dazu betrug der Anteil der Frauen bei den Absolventen in den Ingenieurwissenschaften, Fertigung und Bauwesen in allen Ländern mit Ausnahme von Argentinien, Estland, Island, Polen und Slowenien höchstens ein Drittel. Diese Situation hat sich seit 2000 nur geringfügig verändert, trotz zahlreicher Initiativen der OECD-Länder und der EU zur Verringerung geschlechtsspeziﬁscher Unterschiede. So hat beispielsweise die Europäische Union 2000 das Ziel festgelegt, die Anzahl der Absolventen in Mathematik, Naturwissenschaften und Technik bis 2010 um mindestens 15 Prozent zu steigern sowie eine Verringerung des in diesen Studiengängen bestehenden Ungleichgewichts zwischen den Geschlechtern zu erreichen. Bislang wurden jedoch nur sehr geringe Fortschritte in dieser Hinsicht erzielt. Deutschland, die Schweiz, die Slowakei und Tschechien waren die einzigen vier Länder, in denen der Anteil der Frauen in den Naturwissenschaften zwischen 2000 und 2011 um mindestens 10 Prozentpunkte gestiegen ist. Damit haben sich diese Länder in dieser Hinsicht dem OECD-Durchschnitt angenähert. In den OECD-Ländern ist der Anteil der Frauen in diesen Fachrichtungen geringfügig von 40 Prozent im Jahr 2000 auf 41 Prozent im Jahr 2011 gestiegen – obwohl der Anteil der weiblichen Absolventen über alle Fachrichtungen hinweg in diesem Zeitraum von 54 Prozent auf 58 Prozent gestiegen ist. Der Anteil der Frauen in der Fachrichtung Ingenieurwissenschaften, Fertigung und Bauwesen ist ebenfalls niedrig, er ist jedoch in den letzten 10 Jahren leicht von 23 auf 27 Prozent angestiegen (Tab. A3.3 im Internet). Definitionen Ein Studiengang im Tertiärbereich A, der zu einem ersten Abschluss führt, dauert kumuliert regulär mindestens drei Jahre (vollzeitäquivalent), z. B. beim Bachelorabschluss in vielen englischsprachigen Ländern, dem Diplom in vielen deutschsprachigen Ländern und der licence in vielen französischsprachigen Ländern. Ein Erstabsolvent ist ein Studierender, der innerhalb des Referenzzeitraums zum ersten Mal in einem bestimmten Bildungsbereich – oder im Fall von ISCED 5 in einem Studiengang des Tertiärbereichs A oder B – einen Abschluss erwirbt. Wenn also ein Studierender im Laufe der Jahre mehrere Abschlüsse erwirbt, so wird er oder sie zwar jedes Jahr als Absolvent gezählt, als Erstabsolvent jedoch nur einmal. 70 bildung auf einen blick 2013 Wie viele Studierende werden ein Studium im Tertiärbereich abschließen? Studierende werden als internationale Studierende eingestuft, wenn sie aus ihrem Herkunftsland zwecks Studium in ein anderes Land gekommen sind. Per Deﬁnition gelten sie als Absolventen mit einem Erstabschluss, unabhängig von ihren früher in anderen Ländern erworbenen Abschlüssen. A 3 Netto-Abschlussquoten zeigen den geschätzten prozentualen Anteil einer speziﬁschen Altersgruppe, der im Laufe des Lebens einen Abschluss im Tertiärbereich erwerben wird (ausgehend von den aktuellen Abschlussquoten). Zu einem zweiten Abschluss führende und stärker theoretisch orientierte Studiengänge (z. B. beim Masterabschluss in vielen Ländern) werden getrennt von weiterführenden forschungsorientierten Studiengängen (ISCED 6) in den Tertiärbereich A eingeordnet. Als Absolventen im Tertiärbereich gelten alle Studierenden, die einen Universitätsabschluss, Fachhochschulabschluss oder Abschluss in einem weiterführenden forschungsorientierten Studiengang auf Ebene der Promotion erwerben. Angewandte Methodik Die Daten beziehen sich auf das Studienjahr 2010/2011 und beruhen auf der von der OECD im Jahre 2012 durchgeführten UOE-Datenerhebung zur Bildungsstatistik (weitere Informationen s. Anhang 3 unter www.oecd.org/edu/eag.htm). Die Daten zu den Auswirkungen internationaler Studierender auf die Abschlussquoten im Tertiärbereich basieren auf einer im Dezember 2012 von der OECD durchgeführten speziellen Erhebung. Die Daten zur Entwicklung der Abschlussquoten im Tertiärbereich für die Jahre 1995 und 2000 bis 2004 basieren auf einer speziellen Erhebung, die im Januar 2007 durchgeführt wurde. Um einen von den unterschiedlichen nationalen Abschlussstrukturen unabhängigen Vergleich zu ermöglichen, werden die Abschlüsse in universitären Studiengängen entsprechend ihrer regulären Gesamtdauer untergliedert, mit anderen Worten entsprechend der (gesetzlich oder anders) festgelegten Zahl von Jahren, in denen ein Studiengang abgeschlossen werden kann. Abschlüsse, die nach kurzen Studiengängen mit einer Dauer von weniger als drei Jahren erworben werden, gelten hier nicht als gleichwertig mit einem Abschluss dieses Bildungsbereichs und werden somit bei diesem Indikator nicht berücksichtigt. Zu einem zweiten Abschluss führende Studiengänge werden nach der kumulierten Dauer der zum ersten und zum zweiten Abschluss führenden Studiengänge klassiﬁziert, wobei Absolventen, die bereits über einen ersten Abschluss verfügen, bei der Zählung der Absolventen mit einem Erstabschluss nicht berücksichtigt werden. Außer es ist etwas anderes angegeben, wurden die Abschlussquoten als Netto-Abschlussquoten (d. h. als Summe der Abschlussquoten der einzelnen Altersjahrgänge) berechnet. Für diejenigen Länder, die keine detaillierten Daten bereitstellen konnten, bildung auf einen blick 2013 71 Kapitel A Bildungsergebnisse und Bildungserträge werden die Brutto-Abschlussquoten dargestellt. Zur Berechnung der Brutto-Abschlussquoten gab jedes Land das typische Abschlussalter an (s. Anhang 1). Die Zahl der Absolventen wurde (ohne Berücksichtigung ihres jeweiligen Alters) durch die Bevölkerung im üblichen Abschlussalter geteilt. In vielen Ländern ist es jedoch schwierig, ein typisches Abschlussalter anzugeben, weil die Altersverteilung der Absolventen sehr breit gestreut ist. A 3 Die statistischen Daten für Israel wurden von den zuständigen israelischen Stellen bereitgestellt, die für sie verantwortlich zeichnen. Die Verwendung dieser Daten durch die OECD erfolgt unbeschadet des völkerrechtlichen Status der Golanhöhen, von OstJerusalem und der israelischen Siedlungen im Westjordanland. Weiterführende Informationen OECD (2008), Tertiary Education for the Knowledge Society: Volume 1 and Volume 2, OECD Publishing, http://dx.doi.org/10.1787/9789264046535-en. Tabellen Indikator A3 Tabelle A3.1a: Abschlussquoten und durchschnittliches Abschlussalter im Tertiärbereich (2011) StatLink: http://dx.doi.org/10.1787/888932848362 Tabelle A3.1b: Abschlussquoten im Tertiärbereich von Studierenden, die jünger als das typische Abschlussalter sind (2011) StatLink: http://dx.doi.org/10.1787/888932848381 Tabelle A3.2a: Entwicklung der Abschlussquoten im Tertiärbereich (1995 – 2011) StatLink: http://dx.doi.org/10.1787/888932848400 WEB Table A3.2b: Trends in tertiary graduation rates, by gender (Entwicklung der Abschlussquoten im Tertiärbereich, nach Geschlecht) (2005 – 2011) StatLink: http://dx.doi.org/10.1787/888932848419 WEB Table A3.2c: Trends in net graduation rates at advanced research level (Entwicklung der Netto-Abschlussquoten bei weiterführenden forschungsorientierten Studiengängen) (1995 – 2011) StatLink: http://dx.doi.org/10.1787/888932848438 WEB Table A3.3: Percentage of tertiary qualiﬁcations awarded to women in tertiary-type A and advanced research programmes, by ﬁeld of education (Prozentsatz der von Frauen erworbenen Abschlüsse im Tertiärbereich A und in weiterführenden forschungsorientierten Studiengängen, nach Fachrichtung) (2000, 2011) StatLink: http://dx.doi.org/10.1787/888932848457 72 bildung auf einen blick 2013 Wie viele Studierende werden ein Studium im Tertiärbereich abschließen? Tabelle A3.1a A Abschlussquoten und durchschnittliches Abschlussalter im Tertiärbereich (2011) Summe der Abschlussquoten der einzelnen Altersjahrgänge, nach Geschlecht und Art des Studiengangs 3 Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Durchschnittliches Abschlussalter1 Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Durchschnittliches Abschlussalter1 Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Durchschnittliches Abschlussalter1 Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Durchschnittliches Abschlussalter1 Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Durchschnittliches Abschlussalter1 Tertiärbereich A Weiterführende for(zweiter und weitere schungsorientierte Abschlüsse) Studiengänge Gesamt Tertiärbereich A (erster Abschluss) Durchschnittliches Abschlussalter1 Tertiärbereich A (Erstabschluss) Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Tertiärbereich B (erster Abschluss) Gesamt Tertiärbereich B (Erstabschluss) (1) (4) (5) (6) (9) (10) (11) (14) (15) (16) (19) (20) (21) (24) (25) (26) (29) (30) OECD-Länder Australien2 Österreich Belgien Kanada2 Chile Tschechien Dänemark Estland Finnland Frankreich2 Deutschland Griechenland Ungarn Island2 Irland Israel Italien Japan Korea Luxemburg Mexiko Niederlande Neuseeland Norwegen Polen Portugal Slowakei Slowenien Spanien Schweden Schweiz Türkei Ver. Königreich Vereinigte Staaten OECD-Durchschnitt EU21-Durchschnitt 17 12 m 18 22 5 11 m n m 14 m 7 2 22 m m 25 m m 2 n 29 n 1 n 1 27 18 7 15 17 13 12 11 9 14 11 m 16 22 m 10 m n m m m m 2 21 m m 24 m m m n 21 m m n m 27 m 7 m m m m m m 28 30 m 26 28 27 27 m m m m m 23 38 30 m m m m m 22 m 30 m m m 24 31 24 29 m 24 31 m 28 28 26 14 30 20 23 5 12 19 n 26 14 14 8 2 22 m 1 25 29 m 2 n 35 n 1 n 1 28 18 7 23 17 16 12 14 12 20 13 29 19 23 5 11 19 n 25 14 m 8 2 21 m 1 24 29 m m n 27 n m n m 28 m 7 m 17 15 12 m m 30 32 24 26 28 27 28 28 m m m 26 23 37 30 m m m 25 m 22 m 30 m m m 24 31 24 29 m 24 32 m 28 28 50 35 m 35 24 41 50 m 47 m 31 m 27 60 43 40 32 44 m m 21 42 52 43 58 39 46 37 32 41 32 23 55 39 40 41 33 30 m 33 24 m 45 m m m 29 m m 57 41 m 32 43 m m m 40 42 m 57 39 44 36 m 36 m m m m m m 25 27 m 25 30 28 27 m 28 m 26 m 26 31 25 29 26 m m m 25 24 28 27 26 27 26 26 25 29 29 m 25 m 27 26 60 33 18 37 20 41 50 25 49 36 31 25 31 63 43 40 32 44 48 m 21 45 55 47 58 39 46 37 38 38 28 23 43 39 39 38 43 29 17 35 20 37 47 24 47 32 29 m 29 62 41 39 32 43 47 m m 43 48 46 57 39 44 37 38 37 25 23 37 38 m m 27 27 m 25 28 27 28 25 29 m 26 25 26 31 25 29 26 m 25 m 25 25 27 27 26 27 26 26 27 30 26 25 24 m 27 26 21 10 26 10 6 25 22 12 22 15 5 7 10 24 23 17 24 6 10 m 3 20 18 13 43 23 39 5 8 10 18 3 27 19 17 19 8 9 21 9 6 23 19 12 20 12 4 m 10 22 21 16 m 6 10 m m 19 14 12 43 22 38 5 8 5 14 3 15 17 m m 31 32 m 33 37 29 30 28 32 m 29 m 34 36 32 35 m m 34 m m 27 34 32 m 31 28 35 31 32 31 30 30 m 32 31 1,9 2,1 1,5 1,2 0,2 1,4 2,2 1,3 2,5 1,6 2,7 1,0 0,8 m 1,9 1,4 1,4 1,1 1,4 m 0,2 1,8 1,9 1,9 0,5 1,4 1,9 1,7 1,1 2,8 3,2 0,4 2,4 1,7 1,6 1,7 1,3 1,6 1,1 1,1 0,2 1,3 1,7 1,2 2,2 0,9 2,3 m 0,7 m 1,6 1,4 1,4 0,9 1,3 m m 1,2 1,2 1,7 m 1,3 1,7 1,5 0,9 2,1 1,7 0,4 1,3 1,3 m m 37 33 m 36 37 34 34 36 39 m 33 m 36 m 34 37 32 m 40 m m 32 38 38 m 39 33 35 37 37 33 34 34 m 35 35 Sonst. Saudi-Arabien Südafrika G20-Durchschnitt m m m m m m 8 m m m m m m m m 8 m m m m m m m m m m m 15 6 19 m 1 25 8 5 14 m 6 m m m 25 8 m m m 32 m m m m m m m m m m m m m 18 m m m m m m m m 18 m m m m m m m m m m m 12 28 16 m 12 58 18 6 29 m 28 m m m 57 18 m m m 29 m m 24 m m m m 1 1 n m 1 1 1 4 10 m 1 m m m m 1 m m m m m m m m m m m 0,2 0,4 2,2 m 0,1 0,4 0,1 0,1 1,1 m 0,4 m m m m 0,1 m m m 39 m m 26 m m m m Anmerkung: Die Spalten mit den separaten Abschlussquoten für Männer und Frauen, d. h. die Spalten (2), (3), (7), (8), (12), (13), (17), (18), (22), (23), (27) und (28), stehen im Internet zur Verfügung (s. StatLink unten). Weiterführende Informationen zu den zur Berechnung der (Brutto-/Netto-)Abschlussquoten verwendeten Methoden und des entsprechenden typischen Abschlussalters s. Anhang 1. Unterschiede im Erhebungsbereich der Bevölkerungsdaten und der Absolventendaten bedeuten, dass die Abschlussquoten von Ländern mit einem Nettoabgang von Studierenden wohl zu niedrig angesetzt und von Ländern mit einem Nettozugang von Studierenden wohl zu hoch angesetzt sind. Bei den bereinigten Studienanfängerquoten in den Tabellen A3.1a und A3.1b ist dies weitgehend berücksichtigt. 1. Das „durchschnittliche Abschlussalter“ bezieht sich auf ein durchschnittliches gewichtetes Alter, in der Regel das Alter von Studierenden zu Beginn des Kalenderjahres, sie könnten jedoch schon ein Jahr älter als angegeben sein, wenn der Abschluss gegen Ende des Studienjahres erfolgt. 2. Referenzjahr 2010. Quelle: OECD. Argentinien, China, Indonesien: Statistikinstitut der UNESCO (World Education Indicators Programme). Saudi-Arabien: Observatory on Higher Education. Südafrika: Statistikinstitut der UNESCO. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932848362 Erläuterung der Kennzeichnung fehlender Daten s. Hinweise für den Leser. bildung auf einen blick 2013 73 Kapitel A Abschlussquoten im Tertiärbereich von Studierenden, die jünger als das typische Abschlussalter sind (2011) Summe der Abschlussquoten der einzelnen Altersjahrgänge von bis zu 30-Jährigen im Tertiärbereich A oder B und von bis zu 35-Jährigen bei weiterführenden forschungsorientierten Studiengängen, nach Geschlecht und Art des Studiengangs Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Gesamt Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Weiterführende forschungsorientierte Studiengänge Gesamt Tertiärbereich A (zweiter Abschluss) Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Teritärbereich A (erster Abschluss) Gesamt Tertiärbereich A (Erstabschluss) Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) Tertiärbereich B (erster Abschluss) Gesamt Tertiärbereich B (Erstabschluss) Bereinigte Abschlussquote (ohne internationale/ausländische Studierende) 3 Tabelle A3.1b Gesamt A Bildungsergebnisse und Bildungserträge (1) (4) (5) (8) (9) (12) (13) (16) (17) (20) (21) (24) OECD-Länder Australien1 Österreich Belgien Kanada1 Chile Tschechien Dänemark Estland Finnland Frankreich1 Deutschland Griechenland Ungarn Island1 Irland Israel Italien Japan Korea Luxemburg Mexiko Niederlande Neuseeland Norwegen Polen Portugal Slowakei Slowenien Spanien Schweden Schweiz Türkei Ver. Königreich Vereinigte Staaten OECD-Durchschnitt EU21-Durchschnitt 11 7 m 14 15 4 9 m n m m m 6 m 14 m m m m m 2 n 18 n 1 n 1 16 16 4 m 15 8 m 8 6 8 7 m 13 15 m 8 m n m m m m m 13 m m m m m m n 11 m m n m 16 m 4 m m m m m m 14 8 27 16 16 4 10 13 n m m 13 7 m 14 m m m 25 m 2 n 21 n 1 n 1 16 16 5 m 15 9 m 10 8 9 8 m 15 16 m 8 m n m m m m m 13 m m m 25 m m m 14 m m n m 16 m 5 m m m m m m 42 28 m 32 16 33 41 m 36 m 27 m 23 36 38 29 28 m m m 20 39 37 36 48 33 36 33 29 29 23 m 47 m 33 34 27 23 m 30 16 m 37 m m m 25 m m 34 37 m m m m m m 37 29 m 48 33 36 32 m 25 m m m m m m 48 27 m 33 16 33 39 21 36 m 27 22 26 38 38 29 28 m 46 m 20 41 42 38 48 33 36 33 33 26 24 20 38 m 32 33 33 24 m 31 16 m 37 m 35 m 25 m m 38 36 29 m m 45 m m 39 35 m 48 33 36 32 m 25 22 m m m m m 13 6 m 5 2 20 16 9 12 m 4 m 5 9 13 5 m m 5 m m 17 9 8 m 15 29 2 6 6 11 2 17 m 10 12 3 5 m 5 1 m 14 m 11 m 3 m m 8 12 5 m m 5 m m 16 6 m m 14 29 2 m 2 8 m m m m m 1,0 1,6 m 0,8 0,2 0,7 1,5 0,8 1,1 m 2,2 m 0,5 0,4 1,3 0,7 1,0 m 0,5 m m 1,6 0,9 1,0 m 0,7 1,3 1,2 0,7 1,6 2,6 0,3 1,7 m 1,1 1,2 0,7 1,2 m 0,6 n m 1,1 m 0,9 m 1,9 m m n 1,1 0,7 m m n m m 1,1 0,5 m m 0,6 1,3 1,1 m 1,1 1,3 m m m m m Sonst. Saudi-Arabien Südafrika G20-Durchschnitt m m m m m m m m m m m m m m m m m m m 3 m m 1 m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m 17 m m 12 m m m m m m m m m m m m m m 1 m m 1 m m m m m m m m m m m m m m 0,2 m m 0,1 m m m m m m m m m m m m m Anmerkung: Die Spalten mit den separaten Abschlussquoten für Männer und Frauen, d. h. die Spalten (2), (3), (6), (7), (10), (11), (14), (15), (18), (19), (22) und (23), stehen im Internet zur Verfügung (s. StatLink unten). Weiterführende Informationen zu den zur Berechnung der (Brutto-/Netto-)Abschlussquoten verwendeten Methoden und des entsprechenden typischen Abschlussalters s. Anhang 1. Unterschiede im Erhebungsbereich der Bevölkerungsdaten und der Absolventendaten bedeuten, dass die Abschlussquoten von Ländern mit einem Nettoabgang von Studierenden wohl zu niedrig angesetzt und von Ländern mit einem Nettozugang von Studierenden wohl zu hoch angesetzt sind. Bei den bereinigten Studienanfängerquoten in den Tabellen A3.1a und A3.1b ist dies weitgehend berücksichtigt. 1. Referenzjahr 2010. Quelle: OECD. Indonesien: Statistikinstitut der UNESCO (World Education Indicators Programme). Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932848381 Erläuterung der Kennzeichnung fehlender Daten s. Hinweise für den Leser. 74 bildung auf einen blick 2013 Wie viele Studierende werden ein Studium im Tertiärbereich abschließen? Tabelle A3.2a A Entwicklung der Abschlussquoten im Tertiärbereich (1995 – 2011) Summe der Abschlussquoten der einzelnen Altersjahrgänge, nach Art des Studiengangs 3 Tertiärbereich A (Erstabschluss) Tertiärbereich B (Erstabschluss) 1995 2000 2005 2008 2009 2010 2011 1995 2000 2005 2008 2009 2010 2011 (1) (2) (7) (10) (11) (12) (13) (14) (15) (20) (23) (24) (25) (26) OECD-Länder Australien m 36 50 49 50 50 m m m m 16 16 17 m Österreich 10 15 20 25 29 30 35 m m 8 8 10 12 12 Belgien m m m m m m m m m m m m m m Kanada1 27 27 29 37 36 35 m m m m 29 29 18 m 22 Chile m m m m m m 24 m m m m m m Tschechien 13 14 23 36 38 38 41 6 5 6 5 4 5 5 Dänemark 25 37 46 47 50 50 50 8 10 10 11 11 9 11 m Estland m m m m m m m m m m m m m Finnland 21 40 47 63 44 49 47 34 7 n n n n n Frankreich m m m m m m m m m m m m m m Deutschland1 14 18 20 25 28 30 31 13 11 11 10 14 14 14 Griechenland 14 15 25 m m m m 5 6 11 m m m m Ungarn m m 33 30 31 31 27 m m 4 4 5 6 7 Island 20 33 56 57 51 60 m 10 5 4 4 2 2 m 22 Irland m 30 38 46 47 47 43 m 15 24 26 26 26 Israel m m 35 36 37 37 40 m m m m m m m Italien m 19 41 33 33 32 32 m n 1 1 1 1 m 25 Japan 25 29 37 39 40 40 44 30 30 28 27 26 25 Korea m m m m m m m m m m m m m m Luxemburg m m m m m m m m m m m m m m 2 Mexiko m m 17 18 19 20 21 m m 1 1 1 1 Niederlande 29 35 42 41 42 42 42 m m n n n n n Neuseeland 33 50 51 48 50 47 52 12 17 21 21 24 26 29 Norwegen 26 37 41 41 41 42 43 6 6 2 1 n n n Polen m 34 47 50 50 55 58 m m n n n 1 1 Portugal 15 23 32 45 40 40 39 6 8 9 2 1 n n Slowakei 15 m 30 58 62 49 46 1 2 2 1 1 1 1 Slowenien m m 18 20 27 29 37 m m 24 26 26 26 27 Spanien2 24 29 30 27 27 30 32 2 8 15 14 15 16 18 Schweden 24 28 38 40 36 37 41 m 4 5 6 6 6 7 Schweiz 9 12 27 32 31 31 32 13 14 8 19 19 16 15 Türkei 6 9 11 20 21 23 23 2 m m 13 15 19 17 Vereinigtes Königreich m 42 47 48 48 51 55 m 7 11 12 12 12 13 Vereinigte Staaten 33 34 34 37 38 38 39 9 8 10 10 11 11 12 OECD-Durchschnitt 20 28 34 39 39 39 39 11 9 9 11 11 10 11 OECD-Ländermittel für Länder mit Daten für 1995, 2000 und 2011 20 27 40 11 10 EU21-Durchschnitt 18 27 41 9 7 34 40 39 40 11 8 8 8 8 9 Sonstige G20-Länder Argentinien m m m m m m m m m m m m m m Brasilien m 10 m m m m m m m m m m m m China m m m m m m m m m m m m m m Indien m m m m m m m m m m m m m m Indonesien m m m m m m m m m m m m m m Russische Föderation m m m m m m m m m m m m m m Saudi-Arabien 11 13 18 21 19 20 18 n 3 5 6 6 8 8 Südafrika m m m m m m m m m m m m m m G20-Durchschnitt m m m m m m m m m m m m m m Anmerkung: Die Angaben für die Jahre 2001, 2002, 2003, 2004, 2006 und 2007 sind im Internet verfügbar (s. 1. Unterbrechung der Zeitreihe zwischen 2008 und 2009 aufgrund einer teilweisen Neueinstufung berufsbildender Bildungsgänge in ISCED 2 und ISCED 5B. 2. Unterbrechung der Zeitreihe im Jahr 2008 aufgrund Veränderungen in der Methodik für ISCED 5A. Quelle: OECD. Saudi-Arabien: Observatory on Higher Education. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932848400 Erläuterung der Kennzeichnung fehlender Daten s. Hinweise für den Leser. bildung auf einen blick 2013 75 Wie hoch sind die Erfolgsquoten im Tertiärbereich? A Indikator A4 Wie hoch sind die Erfolgsquoten im Tertiärbereich? 4 Im Durchschnitt der OECD-Länder mit verfügbaren Daten erwerben rund 70 Prozent der Studierenden im Tertiärbereich A mindestens einen ersten Abschluss auf diesem Niveau. Frauen, die sich in einem Studiengang des Tertiärbereichs A eingeschrieben haben, werden eher als Männer am Ende des Studiengangs auch einen Abschluss erlangen: Ihre Erfolgsquote liegt im Durchschnitt 10 Prozentpunkte über derjenigen der Männer. Abbildung A4.1 Anteil der Studierenden, die ein Studium im Tertiärbereich aufnehmen und mindestens einen ersten Abschluss in diesem Bildungsbereich erwerben, nach Geschlecht (2011) Ungarn Frauen Ver. Staaten Schweden2 Norwegen Männer Neuseeland Polen Österreich1 Mexiko Israel1 Portugal Slowakei Ver. Königreich Tschechien1 Niederlande Belgien (fläm.) Türkei Deutschland1 Finnland Spanien1 Frankreich Dänemark Australien1 Gesamt Japan % 100 90 80 70 60 50 40 30 20 10 0 Anmerkung: Einige der Studierenden ohne Abschluss sind womöglich noch eingeschrieben oder haben ihr Studium nicht an der Einrichtung abgeschlossen, an der sie es begonnen haben, wie dies häufig in den Vereinigten Staaten der Fall ist. Einzelheiten zu den für die Berechnung der Erfolgsquoten verwendeten Methoden s. Tabelle A4.1. 1. Nur Tertiärbereich A. 2. Einschließlich Studierender, die einzelne Kurse belegen und womöglich gar nicht beabsichtigen, alle für einen Abschluss erforderlichen Kurse zu belegen. Anordnung der Länder in absteigender Reihenfolge des Anteils der Studierenden, die im Tertiärbereich mindestens einen ersten Abschluss erwerben. Quelle: OECD. Tabelle A4.1. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932846424 Kontext Die Erfolgsquoten im Tertiärbereich können ein Hinweis auf die Efﬁzienz des betreffenden Tertiärbereichs sein, denn sie zeigen auf, wie viele der Studierenden, die ein Studium im Tertiärbereich aufgenommen haben, dieses letztendlich auch erfolgreich abschließen. Niedrige Erfolgsquoten weisen jedoch nicht notwendigerweise auf Inefﬁzienz hin, denn Studierende können ihr Studium aus ganz unterschiedlichen Gründen abbrechen: Sie erkennen, dass sie das falsche Fachgebiet oder den falschen Studiengang gewählt haben; sie erbringen die von den Bildungseinrichtungen vorgegebenen Mindestleistungen nicht, was insbesondere bei einem relativ offenen Zugang bildung auf einen blick 2013 77 Kapitel A A Bildungsergebnisse und Bildungserträge zum Tertiärbereich vorkommt; oder es bieten sich bereits vor Abschluss des Studiums attraktive Beschäftigungsmöglichkeiten an. Studierende sind vielleicht der Ansicht, dass die angebotenen Studiengänge ihren Erwartungen bzw. den Arbeitsmarktanforderungen nicht entsprechen oder dass die Studiengänge länger dauern, als die Studierenden bereit sind, nicht auf dem Arbeitsmarkt aktiv zu sein. Niedrige Erfolgsquoten, d. h. hohe Studienabbruchquoten, können andererseits darauf hinweisen, dass das Bildungssystem den Bedürfnissen der Studierenden nicht gerecht wird. 4 Weitere wichtige Ergebnisse In Neuseeland, Norwegen, Schweden, Ungarn und den Vereinigten Staaten schließen weniger als 60 Prozent der Studierenden, die ein Studium im Tertiärbereich aufnehmen, dieses mit einem ersten Abschluss auf diesem Niveau ab. In Australien, Dänemark, Finnland, Frankreich, Japan und Spanien hingegen sind es mehr als 75 Prozent. Die Erfolgsquoten im Tertiärbereich B sind mit durchschnittlich 61 Prozent gegenüber denen des Tertiärbereichs A etwas niedriger, sie reichen von mindestens 75 Prozent in Deutschland, Japan und der Slowakei bis zu 18 Prozent in den Vereinigten Staaten. Bei Vollzeitstudierenden ist die Wahrscheinlichkeit, dass sie ihr Studium erfolgreich abschließen, höher als bei Teilzeitstudierenden. Die größten Unterschiede zwischen Voll- und Teilzeitstudierenden bestehen in Neuseeland, wo im Tertiärbereich A die Erfolgsquoten für Vollzeitstudierende 34 Prozentpunkte über denjenigen der Teilzeitstudierenden liegen. Studierende verlassen möglicherweise das Bildungssystem ohne Abschluss, weil sich ihnen in einigen Ländern bereits nach einem Studienjahr attraktive Beschäftigungsmöglichkeiten ergeben. In einigen Ländern haben manche ältere Studierende, die ein Studium im Tertiärbereich aufnehmen, gar nicht die Absicht, einen bestimmten Studiengang abzuschließen, sondern wollen vielmehr als Teil des lebenslangen Lernens nur einige Kurse belegen. Es gibt keinen eindeutigen Zusammenhang zwischen der Höhe der von Bildungseinrichtungen des Tertiärbereichs A erhobenen Studiengebühren und den Erfolgsquoten. 78 bildung auf einen blick 2013 Wie hoch sind die Erfolgsquoten im Tertiärbereich? Analyse und Interpretationen A Erfolgsquoten im Tertiärbereich Studierende „schließen ein Studium erfolgreich ab“, wenn sie einen Studiengang im Tertiärbereich A oder B beginnen und mit einem Abschluss eines der beiden Tertiärbereiche abschließen (auch wenn sie hierbei von Tertiärbereich A zu B oder umgekehrt wechseln). Im Durchschnitt der 18 OECD-Länder, für die Daten vorliegen, schließen etwa 32 Prozent der Studierenden keinen Studiengang des Tertiärbereichs erfolgreich ab. 4 In Neuseeland, Norwegen, Schweden, den Vereinigten Staaten und Ungarn erwerben mehr als 40 Prozent der Studienanfänger im Tertiärbereich keinen Abschluss in diesem Bildungsbereich (Tertiärbereich A oder B), im Gegensatz zu den Studierenden in Dänemark, Finnland, Frankreich und Japan, wo dieser Anteil bei unter 25 Prozent liegt. In denjenigen Ländern, für die nur Daten zum Tertiärbereich A verfügbar sind, liegen die Abbruchquoten zwischen 18 Prozent in Australien und 35 Prozent in Österreich (Abb. A4.1). Die Differenz zwischen dem Anteil an Arbeitsplätzen für Hochqualiﬁzierte auf dem Arbeitsmarkt und dem Anteil der Absolventen des Tertiärbereichs (s. Indikator A1) deutet darauf hin, dass die meisten Länder davon proﬁtieren würden, wenn ein größerer Teil der Studierenden das Studium erfolgreich abschließen würde. Eine Steigerung der Absolventenzahlen erfordert je nach Land unterschiedliche Strategien. In den meisten Ländern schließen Vollzeitstudierende ihr Studium eher erfolgreich ab als Teilzeitstudierende. In einigen Ländern haben ältere Studierende jedoch gar nicht die Absicht, einen bestimmten Studiengang abzuschließen, sondern wollen vielmehr als Teil des lebenslangen Lernens einige Kurse belegen. Trotzdem werden diese Studierenden zusammen mit den eher traditionell Vollzeitstudierenden in der Kategorie Studienanfänger im Tertiärbereich erfasst. In Neuseeland zum Beispiel, wo ein Teilzeitstudium eher üblich ist – die Erfolgsquoten für Vollzeitstudierende im Tertiärbereich A liegen um 34 Prozentpunkte über denen der Teilzeitstudierenden –, wird geschätzt, dass etwa jeder fünfte Studierende alle belegten Module erfolgreich abschließt, aber nie die für eine formale Qualiﬁkation erforderliche Gesamtzahl an Modulen belegt. Dadurch gerät eventuell die Erfolgsquote der eher dem traditionellen Modell des Vollzeitstudiums folgenden Studierenden im Tertiärbereich A in den Hintergrund, die 2011 bei 81 Prozent lag (Tab. A4.1 und A4.2). Frauen, die sich in einem Studiengang des Tertiärbereichs A eingeschrieben haben, werden eher als Männer am Ende des Studiengangs auch einen Abschluss erlangen: Ihre Erfolgsquote liegt 10 Prozentpunkte über derjenigen der Männer. Nur in Deutschland, Österreich, Schweden und den Vereinigten Staaten beträgt die Differenz zwischen den Erfolgsquoten von Frauen und Männern weniger als 5 Prozentpunkte. In Finnland, Polen und Tschechien beträgt der geschlechtsspeziﬁsche Unterschied mehr als 15 Prozentpunkte zugunsten der Frauen (Abb. A4.1). bildung auf einen blick 2013 79 Kapitel A Anteil der Studierenden, die ein Studium im Tertiärbereich A aufnehmen und mindesten einen ersten Abschluss in diesem Bildungsbereich erwerben, nach Vollzeit-/Teilzeitstudium (2011) Ungarn Schweden1 Norwegen Polen Ver. Staaten Österreich Neuseeland Israel Portugal Mexiko Frankreich Belgien (fläm.) Slowakei Niederlande Deutschland Tschechien Finnland Spanien Ver. Königreich Dänemark Australien % 100 90 80 70 60 50 40 30 20 10 0 Kein Abschluss im Tertiärbereich A, Studium jedoch erfolgreich im Tertiärbereich B fortgesetzt Erfolgsquoten im Tertiärbereich A Erfolgsquoten im Tertiärbereich A (Studierende im Vollzeitstudium) Erfolgsquoten im Tertiärbereich A (Studierende im Teilzeitstudium) Türkei 4 Abbildung A4.2 Japan A Bildungsergebnisse und Bildungserträge Anmerkung: Einige der Studierenden ohne Abschluss sind womöglich noch eingeschrieben oder haben ihr Studium nicht an der Einrichtung abgeschlossen, an der sie es begonnen haben, wie dies häufig in den Vereinigten Staaten der Fall ist. Einzelheiten zu den für die Berechnung der Erfolgsquoten verwendeten Methoden s. Tabelle A4.1. 1. Einschließlich Studierender, die einzelne Kurse belegen und womöglich gar nicht beabsichtigen, alle für einen Abschluss erforderlichen Kurse zu belegen. Anordnung der Länder in absteigender Reihenfolge des Anteils der Studierenden, die im Tertiärbereich A mindestens einen ersten Abschluss erwerben. Quelle: OECD. Tabellen A4.1 und A4.2. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932846443 Erfolgsquoten im Tertiärbereich A und B Im Durchschnitt der 23 OECD-Länder, für die Daten vorliegen, schließen etwa 30 Prozent der Studierenden des Tertiärbereichs A ihren Studiengang nicht erfolgreich ab. Die Erfolgsquoten sind jedoch in den einzelnen OECD-Ländern sehr unterschiedlich. In Norwegen, Schweden und Ungarn schließen weniger als 60 Prozent der Studierenden im Tertiärbereich A ihr Studium erfolgreich ab, im Gegensatz zu den Studierenden in Australien, Dänemark, Japan und der Türkei, wo dieser Anteil bei mindestens 80 Prozent liegt. Die Erfolgsquoten im Tertiärbereich B liegen mit durchschnittlich 61 Prozent etwas unterhalb denen des Tertiärbereichs A, und auch hier gibt es große Unterschiede zwischen den einzelnen Ländern. Die Erfolgsquoten im Tertiärbereich B reichen von mindestens 75 Prozent in Deutschland, Japan und der Slowakei bis zu nur 18 Prozent in den Vereinigten Staaten (Tab. A4.1). Es gibt viele Diskussionen unter den Entscheidungsträgern in den OECD-Ländern, deren Bildungseinrichtungen niedrige Studiengebühren für Studiengänge im Tertiärbereich A verlangen, ob eine Anhebung der Studiengebühren zu einer Verbesserung der Erfolgsquoten führen würde. Aufgrund der Ergebnisse dieser Erörterungen haben die einzelnen Länder zum Teil ganz unterschiedliche Entscheidungen getroffen. So ist es den Universitäten in Italien aufgrund neuerer gesetzlicher Bestimmungen möglich, bei Überschreiten der regulären Studiendauer für ein Fach von den Studierenden höhere Studiengebühren zu verlangen. Ziel ist, die durchschnittliche Studiendauer im Tertiärbereich zu verkürzen. Einige OECD-Länder haben die Studiengebühren bereits angehoben (und erlassen sie einigen Studierenden bei guten Studienleistungen), um 80 bildung auf einen blick 2013 Wie hoch sind die Erfolgsquoten im Tertiärbereich? einen Anreiz für die Studierenden zu schaffen, das Studium schneller abzuschließen. Im Gegensatz hierzu sind einige Länder bei ihren niedrigen Studiengebühren geblieben, da die politischen Entscheidungsträger in diesen Ländern davon ausgehen, dass höhere Studiengebühren die Studiendauer verlängern könnten, wenn Studierende nebenbei arbeiten müssen, um die Studiengebühren zu bezahlen. A 4 Diese unterschiedlichen Ansätze bei den Studiengebühren könnten der Grund dafür sein, dass sich nur eine schwache Korrelation zwischen den Erfolgsquoten im Tertiärbereich A und der Höhe der von den entsprechenden Bildungseinrichtungen des Tertiärbereichs A erhobenen Studiengebühren ergibt. So verlangen Australien, Japan, die Niederlande und das Vereinigte Königreich Studiengebühren von mehr als 1.500 USDollar (s. Indikator B5) und haben Erfolgsquoten signiﬁkant über dem OECD-Durchschnitt von 70 Prozent. Im Gegensatz dazu verzichten Dänemark und Finnland auf Studiengebühren und bieten in hohem Maße öffentliche Zuschüsse für Studierende, und auch sie haben Erfolgsquoten von mehr als 75 Prozent. Dies ist keineswegs überraschend, denn alle Indikatoren zum Tertiärbereich und vor allem zu den Ertragsraten belegen, dass im Vergleich zu einem Abschluss im Sekundarbereich II ein Abschluss im Tertiärbereich A dem Einzelnen signiﬁkante Vorteile im Hinblick auf das Einkommen (s. Indikator A6) und Beschäftigungsmöglichkeiten (s. Indikator A5) eröffnet. Dies kann ein ausreichend großer Anreiz für Studierende sein, ihr Studium unabhängig von der Höhe der Studiengebühren abzuschließen (s. Indikator A7). Folgen des Nichterlangens eines Abschlusses im Tertiärbereich A Die Aufnahme eines Studiums im Tertiärbereich A, ohne es erfolgreich abzuschließen, bedeutet dann nicht zwangsläuﬁg ein Versagen, wenn es den Studierenden gelingt, ihr Studium im Tertiärbereich B erfolgreich fortzusetzen. Das Gleiche gilt natürlich auch umgekehrt. In Frankreich schließt ein signiﬁkanter Teil der Studierenden das Studium im Tertiärbereich A nicht erfolgreich ab (14 Prozent), sie setzen ihr Studium jedoch erfolgreich im Tertiärbereich B fort. Mit anderen Worten: In Frankreich werden von 100 Studierenden, die ein Studium im Tertiärbereich A beginnen, 68 mindestens einen ersten Abschluss im Tertiärbereich A erwerben, 14 werden zu einem Studiengang im Tertiärbereich B wechseln, 4 werden im Bildungssystem verbleiben, und nur 14 werden das Studium ohne jeglichen Abschluss abbrechen. In Belgien (ﬂäm.), Dänemark, Neuseeland und Schweden wechseln zwischen 3 und 5 Prozent der Studierenden, die keinen Abschluss im Tertiärbereich A erwerben, erfolgreich in den Tertiärbereich B. Diese Umorientierung gibt es auch bei Studierenden, die ursprünglich einen Studiengang im Tertiärbereich B aufgenommen haben: Studierende, die keinen Abschluss im Tertiärbereich B erwerben, wechseln in den Tertiärbereich A – in Dänemark 6 Prozent, in Neuseeland 7 Prozent und in Schweden 6 Prozent. Von diesen Ländern weist nur Neuseeland einen großen Anteil an Studierenden im Tertiärbereich B auf (s. Indikatoren A3 und C3). Das Nichterreichen eines Abschlusses bedeutet nicht, dass die erworbenen Fähigkeiten und Kenntnisse verloren gehen oder auf dem Arbeitsmarkt nicht gewürdigt werden, eine Zeit lang im Arbeitsmarkt zu sein kann sich zudem positiv auf ein späteres Studium auswirken. In Schweden und den Vereinigten Staaten können Studierende den bildung auf einen blick 2013 81 Kapitel A Bildungsergebnisse und Bildungserträge A Kasten A4.1 4 Zusammenhang zwischen Studienanfängerquoten, Abschlussquoten und Erfolgsquoten Diese drei Indikatoren hängen eng zusammen und werden dazu verwendet, die wichtigsten Unterschiede zwischen den Tertiärbereichen in den einzelnen Ländern zu erklären und zu interpretieren. Die Veränderung einer dieser Faktoren kann sich auf die anderen beiden auswirken. Studienanfängerquoten und Abschlussquoten basieren auf der Gesamtbevölkerung, im Gegensatz zu Erfolgsquoten, die anhand der Studienanfänger der typischen Altersgruppe eines bestimmten Bildungsbereichs ermittelt werden. Abbildung A Studienanfänger-, Abschluss- und Erfolgsquoten im Tertiärbereich A (2011) % 120 Studienanfängerquoten Abschlussquoten Erfolgsquoten 100 80 60 40 Ungarn Schweden Norwegen Polen Österreich Ver. Staaten Neuseeland Israel Mexiko Portugal Frankreich1,2 Slowakei Niederlande Deutschland Finnland Tschechien Spanien Ver. Königreich Dänemark Australien1 Türkei 0 Japan 20 1. Referenzzeitraum 2010. 2. Abschlussquoten für einen ersten akademischen Abschluss anstelle von Abschlussquoten für einen Erstabschluss. Anordnung der Länder in absteigender Reihenfolge der Erfolgsquoten in 2011. Quelle: OECD. Tabellen A3.1a, A4.1 und C3.1a. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932846462 Die Deﬁnitionen der Studienanfängerquoten, der Abschlussquoten und der Erfolgsquoten (s. Abschnitt Deﬁnitionen) zeigen den Zusammenhang zwischen diesen Faktoren auf. Tatsächlich entsprechen die Erfolgsquoten nicht einfach der Division der Abschlussquoten durch die Studienanfängerquoten, eine signiﬁkante Veränderung der Studienanfängerquoten oder der Erfolgsquoten wirkt sich jedoch deﬁnitiv auf die Abschlussquoten aus. Ein signiﬁkanter Anstieg/Rückgang der Erfolgsquoten im Tertiärbereich sollte sich unmittelbar auf die Abschlussquoten des Tertiärbereichs auswirken, sofern die Studienanfängerquoten im gleichen Zeitraum stabil bleiben. Dementsprechend kann sich ein signiﬁkanter Anstieg/Rückgang der Studienanfängerquoten direkt auf die Abschlussquoten im Tertiärbereich auswirken, sofern die Erfolgsquoten im Tertiärbereich stabil bleiben. 82 bildung auf einen blick 2013 Wie hoch sind die Erfolgsquoten im Tertiärbereich? Wie bereits zuvor erwähnt, unterscheiden sich jedoch die Erfolgsquoten in den einzelnen OECD-Ländern sehr. Japan rangiert ganz oben mit einer Erfolgsquote von über 90 Prozent der Studierenden, die einen Studiengang im Tertiärbereich A erfolgreich abschließen, während in Schweden ungefähr jeder zweite Studierende den Tertiärbereich verlässt, ohne wenigstens einen ersten Abschluss zu erwerben. A 4 In den Ländern mit niedrigen Erfolgsquoten (wobei zu berücksichtigen ist, dass es viele Gründe dafür geben mag, dass Studierende ihr Studium nicht erfolgreich abschließen) kompensieren hohe Studienanfängerquoten (wie in Neuseeland, Norwegen, Polen und Schweden) diesen Effekt und dienen dazu, die Absolventenzahlen im Vergleich zum OECD-Durchschnitt zu steigern und den Anforderungen des Arbeitsmarktes gerecht zu werden. Diese Länder haben sich dafür entschieden, allen potenziellen Studierenden, z. B. auch internationalen Studierenden oder älteren Studierenden, den Zugang zum Tertiärbereich zu erleichtern, was ihre Platzierung bei beiden Kennzahlen erklärt. Gleichermaßen gleichen in Ländern mit einem begrenzten Zugang zu Studiengängen im Tertiärbereich wie Japan und der Türkei überdurchschnittlich hohe Erfolgsquoten die niedrigeren Studienanfängerquoten aus und steigern so die Abschlussquoten im Vergleich zum OECD-Durchschnitt (Abb. A). Viele Länder haben noch erheblichen Spielraum, um ihre Abschlussquoten zu verbessern. In 11 der 23 Länder mit verfügbaren Daten zum Tertiärbereich A erwerben mehr als 30 Prozent der Studierenden keinen Abschluss in dem ursprünglich begonnenen Bildungsbereich. Blieben die Anfängerquoten in diesen Ländern stabil und/ oder stiegen die Erfolgsquoten auf die gleiche Höhe wie beispielsweise in Japan (rund 90 Prozent), würden die Abschlussquoten stark ansteigen (Abb. A). Tertiärbereich A ohne einen Abschluss verlassen, einige Zeit arbeiten und ihr Studium zu einem späteren Zeitpunkt fortsetzen. Die vor Arbeitsaufnahme bereits abgeschlossenen Module werden später angerechnet. In Ländern mit einem modularen System wie Schweden erhalten Studierende für jeden abgeschlossenen Kurs Credits. Selbst wenn sie ausreichend Kurse für einen Abschluss belegt haben, beantragen sie eventuell keinen formalen Abschluss, da die Credits der einzelnen Kurse vieler Bildungsgänge auf dem Arbeitsmarkt als gleichwertig anerkannt werden. Das Nichterlangen eines Abschlusses ist in den einzelnen Ländern in unterschiedlich starkem Maße ein bildungspolitisches Problem, und Erfolgsquoten sollten daher mit Vorsicht interpretiert werden. Es wird interessant sein zu sehen, ob sich zukünftige Veränderungen der Arbeitsmärkte auf die Anreize für den Einzelnen, ein Studium im Tertiärbereich abzuschließen, auswirken werden. bildung auf einen blick 2013 83 Kapitel A Bildungsergebnisse und Bildungserträge A Definitionen 4 Erfolgsquoten im Tertiärbereich werden deﬁniert als Prozentsatz derjenigen Studienanfänger im jeweiligen Bildungsbereich, die diesen mindestens mit einem ersten Abschluss beenden. Die Erfolgsquote basierend auf echten Kohortendaten wird berechnet als der Anteil der Absolventen (innerhalb von n Jahren) an einer gegebenen Studienanfängerkohorte (prospektiv). Die Erfolgsquote basierend auf Daten einer Querschnitterhebung wird berechnet als das Verhältnis der Zahl der Studierenden, die im Referenzjahr einen ersten Abschluss erwerben, zur Zahl der Studienanfänger im entsprechenden Bildungsbereich vor n Jahren, wobei n der Zahl an Jahren entspricht, die zum Erwerb des Abschlusses in einem Vollzeitstudium erforderlich sind. Netto-Studienanfängerquoten zeigen den geschätzten prozentualen Anteil einer Altersgruppe, der im Laufe des Lebens zum ersten Mal in den Tertiärbereich eintreten wird. Netto-Studienanfängerquoten sind deﬁniert als die Summe der Netto-Studienanfängerquoten für einzelne Altersjahrgänge. Die Gesamtnetto-Studienanfängerquote ist daher die Summe der prozentualen Anteile von Studienanfängern im Tertiärbereich A und B im Alter i im Verhältnis zur Gesamtbevölkerung im Alter i für alle Altersgruppen. Da Daten für einzelne Altersjahrgänge nur für Altersjahrgänge von 15 bis 29 Jahren vorliegen, werden die Netto-Studienanfängerquoten für ältere Studierende anhand von Daten für Altersgruppen von jeweils fünf Jahren geschätzt. Die Studienanfängerquoten sind daher ein Hinweis auf die Zugangsmöglichkeiten zum Tertiärbereich und die Wertschätzung einer Ausbildung im Tertiärbereich (s. Indikator C3). Netto-Abschlussquoten zeigen den geschätzten prozentualen Anteil einer Altersgruppe, der einen Abschluss im Tertiärbereich erwerben wird, ausgehend von den aktuellen Abschlussquoten (s. Anhang 1). Netto-Abschlussquoten werden in der gleichen Weise berechnet wie Studienanfängerquoten. Die Abschlussquoten im Tertiärbereich zeigen auf, in welchem Umfang die Bildungssysteme der einzelnen Länder Erwachsene mit höherwertigen Kenntnissen hervorbringen (s. Indikator A3). Der nicht erfolgreiche Abschluss bezieht sich auf Studierende, die den jeweiligen Bildungsbereich ohne den Erwerb eines ersten Abschlusses verlassen. Als erster Abschluss im Tertiärbereich gilt unabhängig von der Studiendauer jeder Abschluss, der am Ende eines Studiengangs erworben wird, der nicht einen vorherigen erfolgreichen Abschluss im gleichen Bildungsbereich voraussetzt. In manchen Ländern ist die Unterscheidung zwischen Studienunterbrechung und nicht erfolgreichem Abschluss nur schwer möglich. Angewandte Methodik Daten zu den Erfolgsquoten wurden anhand einer speziellen Erhebung im Jahr 2012 ermittelt. In der Hälfte der in Tabelle A4.1 aufgeführten Länder wird die Berechnung der Erfolgsquoten anhand einer Kohortenanalyse durchgeführt, die auf Paneldaten basiert, bei denen einzelne Studierende vom Studienanfang bis zum Abschluss des Studiengangs erfasst werden (echte Kohortenmethode). Die Schätzung für die anderen Länder geht von einem kontinuierlichen Fortschreiten der Studierenden im Tertiärbe- 84 bildung auf einen blick 2013 Wie hoch sind die Erfolgsquoten im Tertiärbereich? reich aus, da zwischen der Kohorte der Absolventen im Referenzjahr und der Kohorte der Studienanfänger n Jahre zuvor Konsistenz erforderlich ist (Querschnittmethode). Diese Annahme könnte die Realität in den einzelnen Ländern jedoch zu stark vereinfachen (s. Anhang 3 unter www.oecd.org/edu/eag.htm). A 4 Die statistischen Daten für Israel wurden von den zuständigen israelischen Stellen bereitgestellt, die für sie verantwortlich zeichnen. Die Verwendung dieser Daten durch die OECD erfolgt unbeschadet des völkerrechtlichen Status der Golanhöhen, von OstJerusalem und der israelischen Siedlungen im Westjordanland. Tabellen Indikator A4 Tabelle A4.1: Erfolgsquoten im Tertiärbereich (2011) StatLink: http://dx.doi.org/10.1787/888932848495 Tabelle A4.2: Erfolgsquoten im Tertiärbereich A, nach Vollzeit-/Teilzeitstudium (2011) StatLink: http://dx.doi.org/10.1787/888932848514 bildung auf einen blick 2013 85 Kapitel A Tabelle A4.1 Erfolgsquoten im Tertiärbereich (2011) Tertiärbereich OECD-Länder Australien Österreich Belgien (fläm.) Kanada Chile Tschechien Dänemark Estland Finnland Frankreich Deutschland Griechenland Ungarn Island Irland Israel Italien Japan Korea Luxemburg Mexiko Niederlande Neuseeland Norwegen Polen Portugal Slowakei Slowenien Spanien Schweden4 Schweiz Türkei Ver. Königreich Ver. Staaten5 OECD-Durchschnitt EU21-Durchschnitt Sonst. G20-Länder Argentinien Brasilien China Indien Indonesien Russische Föd. Saudi-Arabien Südafrika G20-Durchschnitt Querschnitterhebung Querschnitterhebung Echte Kohortendaten m m Echte Kohortendaten (Tertiärbereich A), Querschnitterhebung (Tertiärbereich B) Echte Kohortendaten m Echte Kohortendaten Längsschnitterhebung Echte Kohortendaten (Tertiärbereich A), Querschnitterhebung (Tertiärbereich B) m Querschnitterhebung Frauen Kein Abschluss im Tertiärbereich (4) = 100 – (1) M+F Männer Frauen Kein Abschluss im Tertiärbereich A, Studium jedoch erfolgreich im Tertiärbereich B fortgesetzt M+F Männer Frauen Kein Abschluss im Tertiärbereich B, Studium jedoch erfolgreich im Tertiärbereich A fortgesetzt Tertiärbereich B Tertiärbereich B Männer Tertiärbereich A Tertiärbereich A Erfolgsquoten im Tertiärbereich B (mindestens ein erster Abschluss in einem Studiengang im Tertiärbereich B)3 Referenzjahr(e) für Studienanfänger Erfolgsquoten (mindestens ein erster Abschluss in einem Studiengang im Tertiärbereich A oder B)1 Methode Erfolgsquoten im Tertiärbereich A (mindestens ein erster Abschluss in einem Studiengang im Tertiärbereich A)2 4 M+F A Bildungsergebnisse und Bildungserträge (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) 2005 – 2007 2006 – 2008 2007 – 2008 m m m m 2007 – 2008 m m m m 73 m m m m 66 m m m m 79 m m m m 27 m m 82 65 69 m m 74 63 62 m m 88 67 76 m m m m 4 m m m m 73 m m m m 65 m m m m 79 m m m m 1 m m 2001 2001 72 64 78 28 75 67 83 m 59 49 64 m 2000 – 2001 m 2000 2002 – 2009 2000 – 2001 m a 2002 – 2009 81 m 76 80 78 m 66 m 84 m 83 m 19 m 24 20 80 m 76 68 77 m 66 m 83 m 83 m 3 m a 14 68 m a 73 68 m a m 69 m a m 6 m a 2 1999 – 2002 2008 –20 09 m m m m 75 73 77 a 75 71 77 a m m m m m m m m m m m m m 2009 – 2010 53 48 56 47 48 45 50 m 42 33 47 m m m m m Querschnitterhebung m m Querschnitterhebung Echte Kohortendaten Echte Kohortendaten Echte Kohortendaten Querschnitterhebung Querschnitterhebung Querschnitterhebung m Querschnitterhebung Echte Kohortendaten m Querschnitterhebung Querschnitterhebung Längsschnitterhebung m 2006 – 2007 2009 – 2010 m m m m 2004 – 2006 m m 2007 – 2008 2003 – 2004 2004 1999 – 2000 2006 – 2009 2006 – 2010 2006 –2009 m 2008 – 2009 2002 – 2003 m 2007 – 2008 2007 – 2008 2003 –20 04 m m m m 2008 m m 2009 – 2010 a 2004 1999 – 2000 2008 –20 09 2009 2008 – 2010 m 2007 – 2010 2002 – 2003 m 2009 – 2010 2007 – 2008 2003 –20 04 m m m m 90 m m 66 72 59 59 62 67 71 m m 53 m 75 72 53 68 69 m m m m 87 m m 61 65 56 52 48 59 m m m 53 m 72 m 51 62 61 m m m m 92 m m 72 78 61 64 74 73 m m m 53 m 78 m 54 72 73 m m m m 10 m m 34 28 41 41 38 33 29 m m 47 m 25 28 47 32 31 m m 66 m 91 m m 67 72 66 59 62 67 71 m 78 48 m 88 79 64 70 69 m m 62 m 88 m m 61 65 65 52 48 59 m m 70 48 m 86 m 61 65 62 m m 70 m 95 m m 72 78 67 64 74 73 m m 84 48 m 90 m 67 74 73 m m m m m m m m m 5 m m m m m m 5 m m m m m m m m m m 87 m m 62 m 45 59 64 m 76 m 73 50 m 62 53 18 61 59 m m m m 86 m m 58 m 41 55 46 m 68 m 71 49 m 60 57 18 53 52 m m m m 89 m m 67 m 48 64 68 m 80 m 74 50 m 66 51 18 60 60 m m m m m m m a a 7 m m n m m m 6 m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m Anmerkung: Die Querschnitterhebung bezieht sich auf die Zahl der Absolventen dieser Studiengänge dividiert durch die Zahl der Studienanfänger dieser Studiengänge im Studienanfangsjahr. 1. Die Erfolgsquoten im Tertiärbereich stellen den Anteil derjenigen dar, die ein Studium im Tertiärbereich A oder B aufnehmen und mindestens einen ersten Abschluss in einem Studiengang des Tertiärbereichs A oder B erwerben. 2. Die Erfolgsquoten im Tertiärbereich A stellen den Anteil derjenigen dar, die ein Studium im Tertiärbereich A aufnehmen und mindestens einen ersten Abschluss in einem Studiengang des Tertiärbereichs A erwerben. 3. Die Erfolgsquoten im Tertiärbereich B stellen den Anteil derjenigen dar, die ein Studium im Tertiärbereich B aufnehmen und mindestens einen ersten Abschluss in einem Studiengang des Tertiärbereichs B erwerben. 4. Einschließlich Studierender, die einzelne Kurse belegen und womöglich gar nicht beabsichtigen, alle für einen Abschluss erforderlichen Kurse zu belegen. 5. Die Erfolgsquoten bei Studiengängen im Tertiärbereich A umfassen auch Studierende in einem 4-jährigen Studiengang, die an der Bildungseinrichtung, an dem sie ihr Studium aufnahmen, innerhalb von 6 Jahren einen Abschluss erwarben; die Erfolgsquoten im Tertiärbereich B umfassen auch Studierende aller 2-jährigen Studiengänge, die an der Bildungseinrichtung, an der sie ihr Studium aufnahmen, innerhalb von 3 Jahren einen Abschluss erwarben. Quelle: OECD. Hinweise s. Anhang 3 unter www.oecd.org/edu/eag.htm. StatLink: http://dx.doi.org/10.1787/888932848495 Erläuterung der Kennzeichnung fehlender Daten s. Hinweise für den Leser. 86 bildung auf einen blick 2013 Wie hoch sind die Erfolgsquoten im Tertiärbereich? Tabelle A4.2 A Erfolgsquoten im Tertiärbereich A, nach Vollzeit-/Teilzeitstudium (2011) Methode Referenzjahr(e) für Studienanfänger Tertiärbereich A Anteil der Studienanfänger1 im: Erfolgsquoten im Tertiärbereich A (mindestens ein erster Abschluss in einem Studiengang im Tertiärbereich A) Vollzeitstudium Teilzeitstudium Vollzeitstudium Teilzeitstudium (1) (2) (3) (4) OECD-Länder Australien Querschnitterhebung 2005 – 2007 m m m m Österreich Querschnitterhebung 2006 – 2008 100 m 65 m Belgien (fläm.) Echte Kohortendaten 2007 – 2008 89 11 56 m Kanada m m m m m m Chile m m m m m m Tschechien Echte Kohortendaten 2001 100 m 75 m Dänemark Echte Kohortendaten 2000 – 2001 100 m 80 m Estland m m m m m m Finnland Echte Kohortendaten 2000 m m m m Frankreich Längsschnitterhebung 2002 – 20 09 m m m m Deutschland Echte Kohortendaten 1999 – 2002 m m m m Griechenland m m m m m m Ungarn Querschnitterhebung 2006 – 2007/ 2009 – 2010 63 37 66 32 Island m m m m m m Irland m m m m m m Israel m m m m m m Italien m m m m m m Japan Querschnitterhebung 2004 – 2006 98 2 93 m Korea m m m m m m Luxemburg m m m m m m Mexiko Querschnitterhebung 2007– 2008 100 m 67 m Niederlande Echte Kohortendaten 2003 – 2004 m m m m Neuseeland Echte Kohortendaten 2004 56 44 81 47 Norwegen Echte Kohortendaten 1999 – 2000 m m m m Polen Querschnitterhebung 2006 – 2009 53 47 64 59 Portugal Querschnitterhebung 2006 – 2010 m m m m Slowakei Querschnitterhebung 2006 – 2009 62 38 72 69 Slowenien m m m m m m Spanien Querschnitterhebung 2008 – 2009 m m m m Schweden Echte Kohortendaten 2002 – 2003 m m m m Schweiz m m m m m m Türkei Querschnitterhebung 2007 – 2008 100 a 88 a Vereinigtes Königreich Querschnitterhebung 2007 – 20 08 m m m m Vereinigte Staaten2 Längsschnitterhebung 2003 – 2004 35 81 19 68 OECD-Durchschnitt 83 25 73 m EU21-Durchschnitt 81 33 68 m Sonst.	14,726
https://openalex.org/W3214919200	OpenAlex	Open Science	CC-By	2,021	Cognitive behavioral therapy for irritable bowel syndrome induces bidirectional alterations in the brain-gut-microbiome axis associated with gastrointestinal symptom improvement	Jonathan P. Jacobs	English	Spoken	10,654	19,713	UCLA UCLA Previously Published Works UCLA UCLA Previously Published Works UCLA Previously Published Works Title Cognitive behavioral therapy for irritable bowel syndrome induces bidirectional alterations in the brain-gut-microbiome axis associated with gastrointestinal symptom improvement Permalink https://escholarship.org/uc/item/4zr243z4 Journal Microbiome, 9(1) ISSN 2049-2618 Authors Jacobs, Jonathan P Gupta, Arpana Bhatt, Ravi R et al. Publication Date 2021-12-01 DOI 10.1186/s40168-021-01188-6 Peer reviewed Powered by the California Digital Library University of California eScholarship.org Jacobs et al. Microbiome (2021) 9:236 https://doi.org/10.1186/s40168-021-01188-6 RESEARCH Open Access Cognitive behavioral therapy for irritable bowel syndrome induces bidirectional alterations in the brain-gut-microbiome axis associated with gastrointestinal symptom improvement Jonathan P. Jacobs1,2,3,4†, Arpana Gupta1,2,3†, Ravi R. Bhatt5, Jacob Brawer1,2,3, Kan Gao1,2,3, Kirsten Tillisch1,2,3, Venu Lagishetty2,3,4, Rebecca Firth6, Gregory D. Gudleski6, Benjamin M. Ellingson2,7,8, Jennifer S. Labus1,2,3, Bruce D. Naliboff1,2,3, Jeffrey M. Lackner6† and Emeran A. Mayer1,2,3,9* Bruce D. Naliboff1,2,3, Jeffrey M. Lackner6† and Emeran A. Mayer1,2,3,9* Abstract Background: There is growing recognition that bidirectional signaling between the digestive tract and the brain contributes to irritable bowel syndrome (IBS). We recently showed in a large randomized controlled trial that cognitive behavioral therapy (CBT) reduces IBS symptom severity. This study investigated whether baseline brain and gut microbiome parameters predict CBT response and whether response is associated with changes in the brain-gut-microbiome (BGM) axis. Methods: Eighty-four Rome III-diagnosed IBS patients receiving CBT were drawn from the Irritable Bowel Syndrome Outcome Study (IBSOS; ClinicalTrials.gov NCT00738920) for multimodal brain imaging and psychological assessments at baseline and after study completion. Fecal samples were collected at baseline and post-treatment from 34 CBT recipients for 16S rRNA gene sequencing, untargeted metabolomics, and measurement of short-chain fatty acids. Clinical measures, brain functional connectivity and microstructure, and microbiome features associated with CBT response were identified by multivariate linear and negative binomial models. Results: At baseline, CBT responders had increased fecal serotonin levels, and increased Clostridiales and decreased Bacteroides compared to non-responders. A random forests classifier containing 11 microbial genera predicted CBT response with high accuracy (AUROC 0.96). Following treatment, CBT responders demonstrated reduced functional connectivity in regions of the sensorimotor, brainstem, salience, and default mode networks and changes in white matter in the basal ganglia and other structures. Brain changes correlated with microbiome shifts including Bacteroides expansion in responders. * Correspondence: emayer@ucla.edu * Correspondence: emayer@ucla.edu * Correspondence: emayer@ucla.edu †$CJonathan P. Jacobs and Arpana Gupta are Co-first authors †$CJeffrey M. Lackner and Emeran A. Mayer are Co-senior authors 1G. Oppenheimer Center for Neurobiology of Stress and Resilience, UCLA, Los Angeles, CA, USA 2David Geffen School of Medicine, UCLA, Los Angeles, CA, USA Full list of author information is available at the end of the article 2David Geffen School of Medicine, UCLA, Los Angeles, CA, USA Full list of author information is available at the end of the article * Correspondence: emayer@ucla.edu †$CJonathan P. Jacobs and Arpana Gupta are Co-first authors †$CJeffrey M. Lackner and Emeran A. Mayer are Co-senior authors 1G. Oppenheimer Center for Neurobiology of Stress and Resilience, UCLA, Los Angeles, CA, USA 2David Geffen School of Medicine, UCLA, Los Angeles, CA, USA Full list of author information is available at the end of the article © The Author(s). 2021 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 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. Page 2 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Conclusions: Pre-treatment intestinal microbiota and serotonin levels were associated with CBT response, suggesting that peripheral signals from the microbiota can modulate central processes affected by CBT that generate abdominal symptoms in IBS. CBT response is characterized by co-correlated shifts in brain networks and gut microbiome that may reflect top-down effects of the brain on the microbiome during CBT. Keywords: Cognitive behavioral therapy, Irritable bowel syndrome, Brain-gut-microbiome axis, Neuroimaging, Biomarkers, Outcome prediction We prospectively recruited 84 IBS participants from the parent randomized, controlled, parallel CBT trial (the Irrit- able Bowel Syndrome Outcome Study) [14] and assessed brain resting-state functional connectivity and microstructure at baseline and 2 weeks following CBT treatment. Compos- ition and function of the gut-microbiome were characterized in 34 CBT recipients. By assessing relative abundances and functions of the gut microbiota, multimodal brain imaging, and detailed clinical measures, we aimed to test two major hypotheses: (1) Do baseline brain and/or microbiome param- eters predict treatment outcomes? (2) Is successful treatment response to CBT associated with alteration of brain and microbiome parameters? Introduction Irritable bowel syndrome (IBS) is a common disorder of brain-gut interactions [1, 2] defined by recurrent abdominal pain associated with altered bowel habits in the absence of any gastrointestinal structural, inflammatory, immunologic, or biochemical pathology [3]. Due to the lack of reliable bio- markers capable of IBS classification, symptom improvement and efficacy of treatment are usually assessed by patient- reported measures [4, 5]. The growing recognition of the important role of brain-gut-microbiome (BGM) interactions in regulating GI function, symptom perception, mood, and affect has marked it as a target for therapeutic intervention in IBS [6, 7]. A body of evidence supports the presence of ana- tomical and functional connectivity alterations in brain networks in IBS patients related to emotional arousal, salience assessment, sensorimotor, and brainstem func- tion (reviewed in [8]). In addition, several reports have identified alterations in the relative abundances of mi- crobial taxa in subsets of patients [9, 10], including a study demonstrating an association of specific gut microbial alterations with differences in grey matter volumes of sensory- and salience-related regions [11]. Study oversight Institutional Review Boards at each of two sites ap- proved the protocols governing participants: University at Buffalo (12/18/2012-12/13/2017) and Northwestern University (11/21/2012-10/21/2015). An independent Data Safety Monitoring Board appointed by the National Institute of Diabetes and Digestive and Kidney Diseases monitored the study bi-annually for participant safety, study conduct, and progress. Patients and the public were involved in the development and qualitative testing of clinical materials used in this study. Cognitive behavioral therapy (CBT) is an effective brain-targeted intervention that teaches information processing skills to address psychological factors known to exacerbate abdominal symptoms including maladap- tive coping, intense worry (e.g., catastrophizing, predic- tion error), stress reactivity, and hypervigilance to threat cues [12, 13]. We have recently shown in a large randomized clinical trial that two CBT programs tai- lored for IBS were effective in producing sustained gastrointestinal symptom improvement compared to an IBS education program that controlled for nonspecific effects from undergoing treatment [14]. To the extent that CBT induces symptomatic change through bio- logical pathways, we hypothesized that this occurs by modulating primarily the brain component of the BGM axis, but the effect of these central changes on the rest of the BGM axis and on symptom improvement are not known. We further hypothesized that microbial signals to the brain in the form of neuroactive metabolites in- cluding short-chain fatty acids and serotonin could modulate responsiveness to the biological effects of CBT [15]. Participants IBS participants recruited for this study of biological mechanisms related to IBS symptom improvement after CBT came from the larger parent study, the IBSOS (n= 436), the results of which have been published previously [14]. Detailed inclusion and exclusion criteria have been previously published [14, 16]. In brief, adults aged be- tween 18 and 70 years diagnosed with IBS via Rome III criteria [17] who rated their symptoms as “moderately severe” (i.e., occurred at least twice weekly and caused life interference) were included. Patients were excluded if they had another primary GI disease, malignancy in the past 5 years, major psychiatric comorbidity, were undergoing IBS-targeted psychotherapy, could not com- mit to completing all scheduled visits, reported a gastro- intestinal infection within 2 weeks before evaluation, or used a gut-sensitive antibiotic during the 12 weeks prior to evaluation. Eighty-four participants underwent Page 3 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome model (GLM) framework in CONN, was conducted on all connections of the average 165 × 165 matrix before and after CBT. This was done separately for CBT responders and then for CBT non-responders. Significance was set at α=0.05, and all tests were corrected using the Benjamini- Hochberg procedure at the seed-level. Visualizations were done using circos 0.69 in Linux [20]. neuroimaging and detailed clinical assessment, and thirty-four participants underwent assessment of micro- biome parameters and dietary intake (Diet History Ques- tionnaire II; further description provided in the Supplementary Methods). These participant numbers were based on funding availability, and no selection cri- teria were applied beyond the inclusion/exclusion cri- teria of the parent study. The voxelwise associations between baseline and pre- post treatments changes in diffusion tensor imaging (DTI) parameters were evaluated separately in responders and non-responders to CBT within white matter and subcor- tical structures including the basal ganglia, thalamus, and brainstem. Statistical parametric maps (SPM) were gener- ated using a GLM that accounted for both age and sex. The model involved comparing SPMs that quantified the difference in change in fractional anisotropy (FA) or ap- parent diffusion coefficient (ADC) as a result of treatment in CBT responders and then for non-responders. The GLM was implemented using AFNI (https://afni.nimh.nih. gov). The resulting SPMs were thresholded at a level of significance p<0.05 for each contrast of interest, and a cluster size threshold greater than 250 μL. Trial procedures l bl Eligible patients were randomized to receive 10 sessions of clinic-based CBT or 4 sessions of largely home-based CBT with minimal therapist contact over a 10-week acute phase as previously described [14]. The 10 session version was delivered once a week for 10 weeks, while the 4 ses- sion version was delivered at week 1, week 3, week 5, and week 10. For these analyses, the two CBT protocols were combined as they include the same components and pro- cedures (patient education, self-monitoring, muscle relax- ation, worry control, flexible problem solving, relapse prevention) and have been shown to have efficacy equiva- lence [14, 18]. Fecal 16S rRNA gene sequencing g q g Fresh frozen fecal samples were obtained from 34 pa- tients in this cohort after the baseline screening visit and within 2 weeks after the end of treatment. Participants were provided with home stool collection kits and were asked to store their stool samples in a freezer immedi- ately after collection until pick up by a courier service within 24–48 h of collection for storage at −80°C. Frozen fecal samples were later ground into a coarse powder by mortar and pestle under liquid nitrogen then aliquoted. DNA extraction by bead beating and amplification of the V4 hypervariable region of the 16S rRNA gene were performed according to our published protocol [24, 25]. The library underwent 2×250 sequencing on an Illumina HiSeq 2500 to a mean depth of 107,433 merged se- quences per sample. QIIME 1.9.1 was used to perform quality filtering, merge paired end reads, and cluster se- quences into 97% operational taxonomic units [26]. Tax- onomy was assigned using the Greengenes May 2013 database. Microbial alpha diversity was assessed on data- sets rarefied to equal sequencing depth (37,662) using the Chao1 index of richness, Faith’s phylogenetic diver- sity, and the Shannon index of evenness. Microbial com- position was compared across samples by weighted UniFrac distances and visualized with principal coordi- nates analysis [27]. The significance of differences in mi- crobial composition was assessed using multivariate Adonis with 100,000 permutations [28]. Differential Participants This minimum cluster size is similar to previous studies using cluster- based permutation correction [21–23], while also trying to maximize the sensitivity to DTI changes. Magnetic resonance imaging All participants underwent baseline and post-treatment imaging sessions in a 3T Siemens Prisma MRI Scanner (Siemens, Erlangen, Germany) for a high-resolution T1 structural scan, resting-state functional scan, and diffu- sion tensor image (details of acquisition and processing provided in the Supplementary Methods). Clinical measures Clinical measures were obtained at pre-treatment base- line and 2 weeks after completion of CBT. The 2 week delay was intended to reduce the effects of treatment re- lated processes such as the relationship with the therap- ist and expectation for improvement. Details of the measures used in the larger study have been previously published [18, 19]. The IBS Symptom Severity Scale (IBS-SSS) [4] was used to measure symptom severity in- cluding pain, distention, bowel dysfunction, and quality of life/global well-being. The threshold for clinical im- provement was set a priori at a decrease of 50 points or greater [4]. Participants meeting this criterion were clas- sified as a responder. Additional clinical measures in- cluded IBS self-efficacy, pain unpleasantness, perceived stress, and mood (further description provided in the Supplementary Methods). Results Of the 84 IBS participants who underwent neuroimag- ing, 58 (69%) were classified as CBT responders while 26 (31%) were classified as non-responders based on 50- point or greater decrease on the IBS Symptom Severity Scale post-treatment. Baseline clinical and brain-gut- microbiome parameters were compared between CBT responders and non-responders to determine whether treatment outcome could be predicted. White matter integrity No statistically significant differences in baseline frac- tional anisotropy (FA) or apparent diffusion coefficient (ADC) were observed between CBT responders com- pared to non-responders after adjusting for multiple hy- pothesis testing. Correlations between microbiome changes with brain changes and clinical changes abundance of microbial genera at baseline was deter- mined using multivariate negative binomial mixed models implemented in DESeq2 with bowel habit sub- type and sex as covariates [29]. Longitudinal Adonis and DESeq2 models comparing pre- vs. post-treatment values included participant identifier as a covariate. Partial correlations controlling for age and sex were run between significant brain changes and microbiome or clinical changes in CBT responders compared to non- responders. P values were adjusted for multiple hypoth- esis testing with the Benjamini-Hochberg false discovery rate procedure and significant q values were reported. Brain functional connectivity Responders showed greater baseline connectivity than non-responders between the central autonomic network (right suborbital sulcus) and the emotional regulation network (right inferior frontal sulcus, q=.02; right tri- angular part of the inferior frontal sulcus, q=.02). Clinical measures There were no differences in any of the clinical measures at pre-treatment baseline between responders and non- responders among the 84 subjects who underwent neu- roimaging (Table S1). Fecal metabolomics Fecal aliquots were shipped to Metabolon, Inc. and run as a single batch on their global HD4 metabolomics plat- form. Compounds were identified by comparison of spec- tral features to Metabolon’s proprietary library that includes MS/MS spectral data on more than 3300 purified standards. Results were provided as scaled, imputed abun- dances of 872 known compounds. Stool aliquots addition- ally underwent a targeted LC-MS/MS analysis to measure concentrations of nine short-chain fatty acids. Global metabolomics profiles were compared across participants and visualized in two dimensions using the square root of the Jensen-Shannon divergence and non-metric multidi- mensional scalin g[25]. Stress is a measure used to assess goodness of fit of the reduced dimensions; 0.2 is typically used as a cutoff for suitable fit. Significance of differences between CBT responders and non-responders was deter- mined by Adonis, adjusting for bowel habit and sex. Stat- istical significance of short-chain fatty acids, serotonin, dopamine, and histamine between CBT responders and non-responders at baseline or between pre- vs. post-CBT values was assessed by the non-parametric Mann–Whit- ney U test. Differential abundance testing across the entire dataset was performed on log-transformed data using GLMs that included bowel habit and sex as covariates. Significance was calculated using empirical Bayes moder- ated t statistics implemented in the limma R package [30]. Participant identifier was utilized as a covariate in Adonis and limma analyses of longitudinal data. Random forests classifiers To determine how well baseline microbiome, clinical, or brain data predicted CBT response (responder/non-re- sponder status), random forests classification with 5-fold cross-validation was implemented using the caret R packag e[31]. The accuracy of the resulting classifiers was determined by calculating the area under the receiver op- erating characteristic curve (AUROC). Contribution of each variable to classifier accuracy was assessed by vari- able importance scores, which represent the decrease in classifier accuracy when that variable is permuted. Signifi- cance of differences in accuracy of the derived classifiers was assessed using the bootstrap method of roc.test in the pROC R package. Statistical analysis of neuroimaging data Baseline differences and pre-post CBT treatment changes in resting-state functional connectivity were measured separately in responders and in non-responders. A paired t test controlling for age and sex, using the general linear Jacobs et al. Microbiome (2021) 9:236 Page 4 of 14 Jacobs et al. Microbiome (2021) 9:236 Page 4 of 14 Correlations between microbiome changes with brain changes and clinical changes Metabolomics Fecal targeted and untargeted metabolomics analysis was performed on the 34 subjects to investigate the functional characteristics of the baseline microbiota associated with CBT response. Given the enrichment in responders of Rose- buria, known to be a source of short-chain fatty acids such as butyrate, fecal short-chain fatty acid concentrations were measured. No significant differences were observed in 9 short-chain fatty acids between CBT responders and non- responders (Figure S2). The abundances of microbiome- derived neurotransmitters such as serotonin, norepinephrine, Microbiome composition A subgroup of 34 subjects underwent fecal sampling for microbiome analysis by 16S rRNA gene sequencing. This included 22 CBT responders (65%) and 12 non- responders (35%), who did not differ by clinical mea- sures or baseline stool consistency as assessed by the Bristol Stool Scale (Table S2). Microbial alpha diversity measures of richness, evenness, and phylogenetic diver- sity did not significantly differ between responders and non-responders, while beta diversity analysis demon- strated a significant difference by responder status (p= .016) (Figs. 1, 5A). Responders had increased Roseburia, Lachnobacterium, and unclassified Lachnospiraceae (all Page 5 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Fig. 1 Baseline fecal microbiota and serotonin are associated with CBT response. A, B Principal coordinates (PC) analysis of 16S rRNA sequence data. Each dot represents the baseline microbiome composition of one IBS participant. Color denotes CBT responder status and dots are sized by the fraction of the microbiome comprised of the Bacteroides genus (A) or Clostridiales order (B). C Microbial genera with statistically significant association with CBT responder status (q<.05) are shown. The y axis shows the log2 of the fold change between responders vs. non-responders (NR). Dot size is proportional to mean relative abundance across all samples. D Baseline relative abundances (median scaled) in feces of the neurotransmitters serotonin, dopamine, and histamine. Lines indicate medians. * p<.05 by Mann–Whitney U test. e Non-metric multidimensional scaling analysis (NMDS) (stress=0.20) of global metabolomics profiles. Color denotes CBT responder status. P value calculated by Adonis, adjusting for sex and bowel habit subtype Fig. 1 Baseline fecal microbiota and serotonin are associated with CBT response. A, B Principal coordinates (PC) analysis of 16S rRNA sequence data. Each dot represents the baseline microbiome composition of one IBS participant. Color denotes CBT responder status and dots are sized by the fraction of the microbiome comprised of the Bacteroides genus (A) or Clostridiales order (B). C Microbial genera with statistically significant association with CBT responder status (q<.05) are shown. The y axis shows the log2 of the fold change between responders vs. non-responders (NR). Dot size is proportional to mean relative abundance across all samples. D Baseline relative abundances (median scaled) in feces of the neurotransmitters serotonin, dopamine, and histamine. Lines indicate medians. * p<.05 by Mann–Whitney U test. e Non-metric multidimensional scaling analysis (NMDS) (stress=0.20) of global metabolomics profiles. Color denotes CBT responder status. Microbiome composition P value calculated by Adonis, adjusting for sex and bowel habit subtype members of the Clostridiales order) and decreased Bacteroides, Parabacteroides, and Prevotella (all mem- bers of the Bacteroidales order) compared to non- responders (Fig. 1C). The inverse relationship between the Clostridiales order and Bacteroides (the predomin- ant genus enriched in non-responders) was apparent in the principal coordinates analysis plot, in which a gradient of Clostridiales (with a Bacteroides gradient in the reverse direction) separates baseline samples (Fig. 1A, B). Baseline severity as defined by IBS-SSS was not significantly associated with microbial diver- sity or composition (Figure S1). GABA, dopamine, and histamine were then assessed. Of these, fecal serotonin levels were increased in responders compared to non-responders (p=.03) (Fig. 1C). Norepineph- rine and GABA were not detected, and there was no differ- ence in dopamine or histamine between responders and non-responders. Expanding the analysis to the remaining fecal metabolites, there was no significant overall difference in global metabolomics profile between CBT responders and non-responders (Fig. 1D). Differential abundance testing did not reveal any metabolites with a statistically significant asso- ciation with CBT response after correcting for multiple hy- pothesis testing, though 55 metabolites were nominally significant (Table S3). Responders show Bacteroides expansion and distinct brain changes after CBT compared to non-responders p p Random forest classifiers were created from baseline clinical data, brain features, or microbial taxa and assessed by 5-fold cross-validation. Of these, only the microbiome classifier had high accuracy to predict CBT response (AUROC 0.96). In contrast, a classifier con- structed from demographic traits and baseline clinical measures had AUROC 0.57. Classifiers constructed using differentially abundant resting-state connections, FA, or ADC clusters had AUROC of 0.66, 0.51, and 0.49, respectively; when all brain datasets were com- bined, the resulting classifier had AUROC of 0.67. The increased accuracy of the microbiome classifier was highly significant by bootstrap analysis (p=6×10−5 and p=.0001 compared to the clinical/demographic and combined brain classifiers, respectively; there was no significant difference between the clinical/demographic and brain classifiers) (Fig. 2A). The taxa contributing most to microbiome classifier accuracy were unclassi- fied Erysipelotrichaceae, unclassified Lachnospiraceae, Roseburia, and Bacteroides (Fig. 2B). Diet We assessed baseline dietary intake for the 34 subjects who underwent fecal sampling using a food frequency questionnaire. CBT responders had lower carbohydrate intake, higher total fat and monounsaturated fat intake, and a trend towards increased fiber intake compared to non-responders (Table S4). However, there were no sig- nificant differences in intake of 32 food groups or 122 individual nutrients, including tryptophan. Page 6 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Classifiers to predict CBT response Responders show Bacteroides expansion and distinct brain changes after CBT compared to non-responders Post-treatment changes in clinical, neuroimaging, and microbiome parameters were compared between re- sponders and non-responders to characterize the differ- ential effects of CBT on the brain-gut-microbiome axis in responders. Responders show Bacteroides expansion and distinct brain changes after CBT compared to non-responders Post-treatment changes in clinical, neuroimaging, and microbiome parameters were compared between re- sponders and non-responders to characterize the differ- ential effects of CBT on the brain-gut-microbiome axis in responders. Clinical changes post-CBT Following the intervention, responders among the 84 subjects with neuroimaging data had a significant de- crease in abdominal pain unpleasantness (Gracely, p= 1×10−6), IBS symptom intensity (Gracely, p=4×10−10), negative mood ratings (POMS, p=.002), and perceived stress (PSS, p=1×10−5) compared to non-responders (Table S5). In addition, responders had a significant in- crease in IBS self-efficacy (p=2×10−14) and positive mood ratings (p=.0001). Although not as high and widespread as in the responders, the non-responders also had a sig- nificant increase in IBS self-efficacy (p=.001), and a de- crease in IBS symptom intensity (p=.03). Fig. 2 Classifiers derived from baseline fecal microbiota profiles outperformed those based on clinical/demographic and neuroimaging data to predict CBT response. A Receiver operating characteristic curves of random forest classifiers for CBT response constructed from differentially abundant microbial genera, baseline clinical/demographic data (left panel), or brain data (right panel). The 95% confidence intervals are represented as colored regions surrounding these curves (blue=microbiome, red=clinical/demographics or brain). B Importance scores for the 11 microbial genera in the random forests classifier Fig. 2 Classifiers derived from baseline fecal microbiota profiles outperformed those based on clinical/demographic and neuroimaging data to predict CBT response. A Receiver operating characteristic curves of random forest classifiers for CBT response constructed from differentially abundant microbial genera, baseline clinical/demographic data (left panel), or brain data (right panel). The 95% confidence intervals are represented as colored regions surrounding these curves (blue=microbiome, red=clinical/demographics or brain). B Importance scores for the 11 microbial genera in the random forests classifier Fig. 2 Classifiers derived from baseline fecal microbiota profiles outperformed those based on clinical/demographic and neuroimaging data to predict CBT response. A Receiver operating characteristic curves of random forest classifiers for CBT response constructed from differentially abundant microbial genera, baseline clinical/demographic data (left panel), or brain data (right panel). The 95% confidence intervals are represented as colored regions surrounding these curves (blue=microbiome, red=clinical/demographics or brain). B Importance scores for the 11 microbial genera in the random forests classifier Page 7 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Clinical changes post-CBT Microbiome Table 1 CBT-related changes in resting-state functional connectivity in responders and non-responders Analysis Unit Table 1 CBT-related changes in resting-state functional connectivity in responders and non-responders Analysis Unit t statistic df p value q value Cohen’s d Interpretation Responders to CBT Right anterior transverse temporal gyrus (Heschl’s Gyrus) —right anterior insula (anterior segment of the circular sulcus of the insula) −4.07 117 .0001 .014 −0.75 Decrease after CBT Left anterior insula (inferior segment of the circular sulcus of the insula)—right planum temporale −4.00 117 .0001 .018 −0.74 Decrease after CBT Left anterior insula (left anterior segment of the circular sulcus of the insula)—right aMCC −3.80 117 .0002 .038 −0.70 Decrease after CBT Right amygdala—right lateral aspect of the superior temporal gyrus −3.79 117 .0002 .040 −0.70 Decrease after CBT Right anterior insula (anterior segment of the circular sulcus of the insula)—right planum polare of the superior temporal gyrus −3.59 117 .0005 .040 −0.66 Decrease after CBT Brainstem—right planum temporale −3.44 117 .0008 .044 −0.64 Decrease after CBT Brainstem—right anterior transverse temporal gyrus (Heschl’s Gyrus) −3.40 117 .0009 .044 −0.63 Decrease after CBT Brainstem—left lateral aspect of the temporal gyrus −3.36 117 .001 .044 −0.62 Decrease after CBT Right anterior insula (anterior segment of the circular sulcus of the insula)—left anterior transverse temporal gyrus (Heschl’s gyrus) −3.42 117 .0009 .048 −0.63 Decrease after CBT Non-responders to CBT Right dPCC—right vPCC −3.74 117 .0003 .047 −0.69 Decrease after CBT Right inferior occipital gyrus and sulcus—right middle frontal gyrus −3.74 117 .0003 .047 −0.69 Decrease after CBT Abbreviations: df degrees of freedom, CBT cognitive behavioral therapy, aMCC anterior mid-cingulate cortex, dPCC dorsal posterior cingulate cortex, vPCC ventral posterior cingulate cortex Table 1 CBT-related changes in resting-state functional connectivity in responders and non-responders Non-responders to CBT Right dPCC—right vPCC Brain functional connectivity changes post-CBT Microbiome and metabolomics changes post CBT Responders among the 34 subjects that underwent fecal sampling were found after CBT to have statisti- cally significant post-treatment decreases in richness and phylogenetic diversity, whereas non-responders analyzed separately had no change in microbial diversity after treatment (Fig. 5A). Responders also demonstrated a highly significant shift in microbial composition (p=2×10−5) whereas non-responders showed no overall change in microbial composition (Fig. 5B). Differential abundance testing revealed five genera that were enriched in responders after CBT, including Bacteroides, Odoribacter, Parabacteroides, Anaerotruncus, and unclassified S24-7 (Fig. 5C). Of these, Bacteroides was the most abundant and had the greatest magnitude of change (2.5 fold increase, q=4×10−5), resulting in conversion of many patients to Bacteroides-predominant microbiota by the end of treatment (Fig. 5B). Metabolomics profiles did not dif- fer between baseline and end of treatment for both groups, and no differentially abundant metabolites or short-chain fatty acids were identified in CBT re- sponders after treatment when adjusting for multiple hypothesis testing (Table S7, Figure S2). Following CBT, responders showed a decrease in connectivity between multiple regions associated with specific brain net- works, including the sensorimotor, default mode, salience, and emotion regulation networks (Table 1, Fig. 3). They also exhibited decreases in connectivity between regions associated with the brainstem and default mode and sensorimotor net- works (all q<.05). Non-responders showed a decrease in con- nectivity between two links, one within the posterior cingulate (default mode network, q=.047) and one between the middle frontal gyrus (central executive network) and inferior occipital gyrus and sulcus (occipital lobe, q=.047). No increases in connectivity were observed for anyone undergo- ing CBT. Changes in brain parameters in CBT responders correlate with clinical outcomes Changes in brain parameters in CBT responders correlate with clinical outcomes CBT responders among the 34 subjects that under- went microbiome analysis did not have any significant changes in dietary macronutrient content or intake of specific food groups and nutrients as assessed by a food frequency questionnaire administered 2 weeks after completion of CBT (Table S8). There were no significant changes in stool consistency as assessed by the Bristol Stool Scale in either responders or non- responders (Figure S3). White matter integrity changes post-CBT g y g p A significantly higher change in FA was observed in CBT non-responders within the left inferior longitudinal fasciculus (Fig. 4A; cluster size = 327 μL). Additionally, we observed a significantly higher ADC in CBT re- sponders bilaterally within areas encompassing the basal ganglia and anterior thalamus (Fig. 4B; left cluster size = 277 μL, right cluster size = 258 μL) as well as the isth- mus of the corpus callosum (Fig. 4C; cluster size = 392 μL) in CBT responders compared with non-responders. Page 8 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Fig. 3 Changes in functional connectivity in responders and non-responders to CBT. A Connectograms demonstrating pair-wise connectivity differences in responders and non-responders to CBT. Significant decreases in connectivity between brain regions are denoted by blue lines connecting the regions (color intensity indicates magnitude of effect). There were no significant increases in connectivity. SMN: sensorimotor network, BG: basal ganglia, DMN: default mode network, SAL: salience network, ERN: emotion regulation network, CAN: central autonomic network, CEN: central executive Network, OCC: occipital/visual network. B Regions that significantly differed between responders and non-responders to CBT. Responders to CBT: ACirIns (anterior insula/anterior segment of the circular sulcus of the insula), MPosCgG/S (anterior mid-cingulate cortex), InfCirIns (anterior insula/inferior segment of the circular sulcus of the insula), SupTGLp (lateral aspect of the superior temporal gyrus), HG (Heschl’s gyrus), TPI (planum temporale), BSt (brainstem). Non-responders to CBT: MFG (middle frontal gyrus), PosDCgG (dorsal posterior cingulate cortex), PosVCgG (ventral posterior cingulate cortex), InfOcG/S (inferior occipital gyrus and sulcus) Fig. 3 Changes in functional connectivity in responders and non-responders to CBT. A Connectograms demonstrating pair-wise connectivity differences in responders and non-responders to CBT. Significant decreases in connectivity between brain regions are denoted by blue lines connecting the regions (color intensity indicates magnitude of effect). There were no significant increases in connectivity. SMN: sensorimotor network, BG: basal ganglia, DMN: default mode network, SAL: salience network, ERN: emotion regulation network, CAN: central autonomic network, CEN: central executive Network, OCC: occipital/visual network. B Regions that significantly differed between responders and non-responders to CBT. Responders to CBT: ACirIns (anterior insula/anterior segment of the circular sulcus of the insula), MPosCgG/S (anterior mid-cingulate cortex), InfCirIns (anterior insula/inferior segment of the circular sulcus of the insula), SupTGLp (lateral aspect of the superior temporal gyrus), HG (Heschl’s gyrus), TPI (planum temporale), BSt (brainstem). White matter integrity changes post-CBT Non-responders to CBT: MFG (middle frontal gyrus), PosDCgG (dorsal posterior cingulate cortex), PosVCgG (ventral posterior cingulate cortex), InfOcG/S (inferior occipital gyrus and sulcus) Fig. 3 Changes in functional connectivity in responders and non-responders to CBT. A Connectograms demonstrating pair-wise connectivity differences in responders and non-responders to CBT. Significant decreases in connectivity between brain regions are denoted by blue lines connecting the regions (color intensity indicates magnitude of effect). There were no significant increases in connectivity. SMN: sensorimotor network, BG: basal ganglia, DMN: default mode network, SAL: salience network, ERN: emotion regulation network, CAN: central autonomic network, CEN: central executive Network, OCC: occipital/visual network. B Regions that significantly differed between responders and non-responders to CBT. Responders to CBT: ACirIns (anterior insula/anterior segment of the circular sulcus of the insula), MPosCgG/S (anterior mid-cingulate cortex), InfCirIns (anterior insula/inferior segment of the circular sulcus of the insula), SupTGLp (lateral aspect of the superior temporal gyrus), HG (Heschl’s gyrus), TPI (planum temporale), BSt (brainstem). Non-responders to CBT: MFG (middle frontal gyrus), PosDCgG (dorsal posterior cingulate cortex), PosVCgG (ventral posterior cingulate cortex), InfOcG/S (inferior occipital gyrus and sulcus) Brain functional connectivity Alterations in neuroimaging parameters following CBT were correlated with microbiome shifts in subjects who underwent fecal sampling. In responders, the increases in the abundance of two taxa (Bacteroides, r(27)=−.63, q =.022; unclassified S24-7, r(27)=−.55, q=.041) were nega- tively associated with reduced resting-state connectivity between the brainstem and a region in the temporal net- work (Table 3). Although responders and non-responders to CBT did not differ on any clinical or behavioral parameters pre- treatment, minor baseline differences were observed in functional connectivity and significant baseline differences were observed in the relative abundances of gut microbes. This suggests that CBT responders have pre-existing dif- ferences in central and peripheral components of the brain-gut-microbiome axis relative to non-responders that confer increased sensitivity to the effects of CBT on GI symptomology. Compared to CBT non-responders, Discussion We demonstrate that a positive clinical response in CBT-treated IBS patients is associated with changes in functional and structural connectivity of brain networks, as well as changes in gut microbiota, compared to CBT- treated patients who did not achieve clinical response. Additionally, we found that a classifier including 11 mi- crobial genera was able to predict treatment outcome with high accuracy. To our knowledge, this is the first demonstration of an association between treatment out- comes and changes in brain and gut microbiota for any IBS therapy (e.g., pharmacological, dietary). These find- ings support the concept that even though CBT is con- sidered a psychological treatment that teaches cognitive skills for remediating perceptual biases, the symptom changes it induces may occur via modulation of brain- gut-microbiome interactions which influence IBS patho- physiology and visceral symptom generation. Brain functional connectivity (Table 2) In responders, decreased connectivity between key nodes of the salience network (right anterior mid-cingulate cortex and left anterior insula) was associated with higher positive mood ratings (POMS; r(49)=.29, q=.036). Reduced connectivity between the brainstem and left su- perior temporal gyrus was associated with higher positive mood (POMS; r(49)=.36, q=.036) and lower negative Page 9 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Fig. 4 CBT responders had distinct changes in white matter integrity compared to non-responders. A Colored areas indicate regions within the left inferior longitudinal fasciculus that had a significant change in FA after CBT. Color corresponds to relative difference in FA change between CBT responders and non-responders. B, C Colored areas indicate regions of the bilateral basal ganglia and anterior thalamus (B) and isthmus of the corpus callosum (C) that had a significant change in ADC after CBT. Color corresponds to relative difference in ADC change between CBT responders and non-responders Fig. 4 CBT responders had distinct changes in white matter integrity compared to non-responders. A Colored areas indicate regions within the left inferior longitudinal fasciculus that had a significant change in FA after CBT. Color corresponds to relative difference in FA change between CBT responders and non-responders. B, C Colored areas indicate regions of the bilateral basal ganglia and anterior thalamus (B) and isthmus of the corpus callosum (C) that had a significant change in ADC after CBT. Color corresponds to relative difference in ADC change between CBT responders and non-responders mood (POMS; r(49)=−.32, q=.042). Decreased connectiv- ity between the right anterior INS and right superior temporal gyrus was associated with lower unpleasantness rating of abdominal pain (Gracely; r(49)=−.33, q=.036). White matter integrity (Table 2) In responders, CBT-associated decreases in FA values in the inferior longitudinal fasciculus were significantly associated with increases in positive affect (PANAS, positive, q=.040), and higher ADC in the brainstem was associated with higher positive mood scores (POMS positive, q=.040). White matter integrity No statistically significant correlations were observed be- tween the microbiome and white matter integrity in responders. Jacobs et al. Microbiome (2021) 9:236 Page 10 of 14 Jacobs et al. Microbiome Fig. 5 CBT responders have altered intestinal microbiome composition after CBT characterized by Bacteroides expansion. A Fecal microbial alpha diversity is shown for CBT responders and non-responders (NR) at baseline (PRE) and after CBT (POST). Three metrics are used: Chao1 index (richness), Faith’s phylogenetic diversity (PD), and Shannon index (richness and evenness). * p<.05. B Principal coordinates analysis of 16S rRNA sequence data before and after CBT, stratified by CBT response status. Each dot represents a sample, colored by time point (red=baseline, blue=post-CBT) and sized by Bacteroides abundance. Arrows connect samples from the same participants, with post-treatment indicated by the arrowhead. P values calculated by Adonis, adjusting for participant. C Microbial genera with statistically significant association with CBT responder status (q<.05) are shown. The y axis shows the log2 of the fold change between responders vs. non-responders. Dot size is proportional to mean relative abundance across all samples Fig. 5 CBT responders have altered intestinal microbiome composition after CBT characterized by Bacteroides expansion. A Fecal microbial alpha diversity is shown for CBT responders and non-responders (NR) at baseline (PRE) and after CBT (POST). Three metrics are used: Chao1 index (richness), Faith’s phylogenetic diversity (PD), and Shannon index (richness and evenness). * p<.05. B Principal coordinates analysis of 16S rRNA sequence data before and after CBT, stratified by CBT response status. Each dot represents a sample, colored by time point (red=baseline, blue=post-CBT) and sized by Bacteroides abundance. Arrows connect samples from the same participants, with post-treatment indicated by the arrowhead. P values calculated by Adonis, adjusting for participant. C Microbial genera with statistically significant association with CBT responder status (q<.05) are shown. The y axis shows the log2 of the fold change between responders vs. non-responders. Dot size is proportional to mean relative abundance across all samples findings suggest that IBS-characteristic brain changes were not more pronounced in CBT responders versus CBT non-responders at baseline and that the observed brain differences between the two groups were only more pro- nounced after the CBT intervention [21, 32, 33]. responders at baseline showed greater functional connect- ivity between regions of the central autonomic network and the emotional regulation network. White matter integrity Microbiome Table 3 Associations between resting-state functional connectivity after CBT with abundance of gut microbes in responders Changes in functional connectivity after CBT are associated with abundance of gut microbes in responders Microbial genera Connectivity—dyad r df p value q value Bacteroides Brainstem—left lateral aspect of the temporal gyrus −.63 27 .011 .022 Unclassified S24-7 Brainstem—right planum temporale −.55 27 .041 .041 Abbreviations: r partial correlation (controlling for age and sex), df degrees of freedom, p p value, CBT cognitive behavioral therapy Table 3 Associations between resting-state functional connectivity after CBT with abundance of gut microbes in responders Changes in functional connectivity after CBT are associated with abundance of gut microbes in responders Microbial genera Connectivity—dyad r df p value q value Bacteroides Brainstem—left lateral aspect of the temporal gyrus −.63 27 .011 .022 Unclassified S24-7 Brainstem—right planum temporale −.55 27 .041 .041 Abbreviations: r partial correlation (controlling for age and sex), df degrees of freedom, p p value, CBT cognitive behavioral therapy While there is currently no agreement on the existence of IBS characteristic alterations in the gut microbiota, evidence for possible microbial-based subgroups has been reported [9, 10]. The current findings add to this concept, demonstrating that microbiota could identify subsets of IBS-patients with differential responsiveness to CBT. Responders had increased levels of several members of the Clostridiales order (Roseburia, Lachno- bacterium, unclassified Lachnospiraceae) and decreased levels of members of the Bacteriodales order (Bacter- oides, Parabacteroides, Prevotella) compared to non- responders, resulting in gradients of Clostridiales and Bacteroides separating the baseline samples. Strikingly, a classifier using 11 differentially abundant genera showed high accuracy to predict CBT response from baseline microbiota. This contrasted with the weak predictions that could be made from clinical or neuroimaging data and suggests that microbial composition could serve as an accurate biomarker for biological pathways in IBS symptom generation that are modulated by CBT if these findings are validated in an independent cohort. Diet is one potential factor contributing to baseline microbial differences between CBT responders and non- responders. In our study, we found that responders re- ported decreased fraction of energy intake from carbohy- drates and increased fraction from unsaturated fats. Dietary pattern, however, was not affected by treatment and is therefore unlikely to explain changes in clinical and biological parameters. statistically significant association with CBT response after correcting for multiple hypothesis testing. White matter integrity However, 55 metabolites were nominally significant, including four—delta-tocopherol, gamma-tocopherol/beta-tocoph- erol, chenodeoxycholate, and m-tyramine—related to metabolites previously shown to induce serotonin release by cultured Enterochromaffin cells in vitro [34]. A recent study by E. Hsiao’s group in mouse models demon- strated that different interventions aimed at increasing intestinal serotonin result in an increase of spore- forming Clostridia, including Clostridiales and Lachnos- piraceae [37]. These findings, together with the earlier study from their lab, support a circular interaction between Clostridiales in the gut, host epithelial serotonin production and release, and serotonin-induced increase in Clostridia abundance, in which Clostridiales promote their own community membership in the gut microbiota. Following treatment, and similar to the results ob- tained in the parent study, responders showed highly significant improvements in several clinical parameters, including IBS symptom severity and its constituent pro- cesses (abdominal pain, life satisfaction, bowel dissatis- faction). These clinical improvements were accompanied by functional and structural brain changes as responders showed a reduction in the connectivity between multiple cortical networks, including the sensorimotor, default mode, salience, and emotion regulation networks. A pre- vious report on the effectiveness of CBT in patients with multiple chronic pain conditions had shown a similar re- duction in the functional connectivity between regions of the default mode and emotion regulation networks which was associated with an increase in functional con- nectivity between the basal ganglia network and the right somatosensory cortex [38]. Similar to our study, they showed that these brain changes were associated with improvements in clinical and behavioral measures. In the current study, a reduction in the connectivity within the salience network and a reduction in brainstem con- nectivity with the left superior temporal gyrus were both associated with more positive mood ratings and reduced negative affective ratings of IBS symptoms. CBT re- sponders also showed significant changes in white mat- ter integrity, with greater change in ADC within the basal ganglia and anterior thalamic regions, and in- creased ADC values in white matter tissue near the right The strong association of the baseline microbiome with CBT response suggests that signals from the gut microbiome could potentiate or suppress the effects of CBT on target brain regions. Clostridiales encompasses many spore-forming microbes which have previously been shown to induce intestinal serotonin synthesis and release, representing a potential link between the gut microbiome and CBT responsiveness [34]. White matter integrity Based on previ- ously reported functional and anatomical connectivity brain differences between IBS and healthy controls, these between changes in resting-state functional connectivity and DTI after CBT with clinical measures in Table 2 Associations between changes in resting-state functional connectivity and DTI after CBT with clinical measures in responders Changes in functional connectivity after CBT are associated with clinical measures in responders Dyad with reduced connectivity Clinical measure r df p value q value Right aINS—right PoPl Unpleasantness rating of IBS symptoms −.33 49 .02 .036 Left aINS—right aMCC POMS positive score .29 49 .039 .036 Brainstem—left SupTGLp POMS positive score .36 49 .009 .036 Brainstem—left SupTGLp POMS negative score −.32 49 .022 .042 Changes in DTI after CBT are associated with clinical measures in responders DTI measure—cluster Clinical measure r df p value q value FA—inferior longitudinal fasciculus PANAS positive affect .38 41 .012 .04 ADC—brainstem POMS—positive score .41 42 .006 .03 Abbreviations: r partial Pearson’s correlation (controlling for age and sex), df degrees of freedom, p p value, CBT cognitive behavioral therapy, IBS irritable bowel syndrome, aMCC anterior mid-cingulate cortex, aINS anterior insula, SupTGLp lateral aspect of the superior temporal gyrus, PoPl planum polare of the superior temporal gyrus, FA fractional anisotropy, ADC apparent diffusion coefficient, POMS Profile of Mood States, PANAS Positive and Negative Affect Schedule Abbreviations: r partial Pearson’s correlation (controlling for age and sex), df degrees of freedom, p p value, CBT cognitive behavioral therapy, IBS irritable bowel syndrome, aMCC anterior mid-cingulate cortex, aINS anterior insula, SupTGLp lateral aspect of the superior temporal gyrus, PoPl planum polare of the superior temporal gyrus, FA fractional anisotropy, ADC apparent diffusion coefficient, POMS Profile of Mood States, PANAS Positive and Negative Affect Schedule Abbreviations: r partial Pearson’s correlation (controlling for age and sex), df degrees of freedom, p p value, CBT cognitive behavioral therapy, IBS irritable bowel syndrome, aMCC anterior mid-cingulate cortex, aINS anterior insula, SupTGLp lateral aspect of the superior temporal gyrus, PoPl planum polare of the superior temporal gyrus, FA fractional anisotropy, ADC apparent diffusion coefficient, POMS Profile of Mood States, PANAS Positive and Negative Affect Schedule Page 11 of 14 Jacobs et al. Microbiome (2021) 9:23 Jacobs et al. Acknowledgements We thank Cathy Liu and the members of the IBSOS Research Team at NIDDK, the University at Buffalo, New York University, and Northwestern for their support of various aspects of this study. Supplementary Information pp y The online version contains supplementary material available at https://doi. org/10.1186/s40168-021-01188-6. pp y The online version contains supplementary material available at https://doi. org/10.1186/s40168-021-01188-6. Additional file 2. Short-chain fatty acids. Additional file 3. Additional file 2. Short-chain fatty acids. Additional file 3. White matter integrity Fecal metabo- lomics analysis showed increased baseline serotonin levels in responders, consistent with a greater tonic lu- minal release of serotonin into the gut lumen or poten- tially serotonin production by luminal bacteria [35]. Acute tryptophan depletion has previously been shown to induce IBS-like symptoms and brain changes in healthy controls, supporting a role of serotonin as a modulator of the BGM axis [36]. Differential abundance testing did not reveal any other metabolites with a Page 12 of 14 Page 12 of 14 Page 12 of 14 Jacobs et al. Microbiome (2021) 9:236 somatosensory regions, compared to non-responders. This CBT-associated normalization of microstructural brain changes in the basal ganglia and thalamic areas were associated with increase in positive mood and posi- tive affect. The IBS patients with improved symptoms following CBT had decreased connectivity between the bilateral Heschl’s gyrus and the anterior insula, a key hub of the salience network. The Heschl’s gyrus is known as the primary auditory cortex which would be highly active during a resting-state fMRI scan due to the noise present in the scanner. Lower connectivity with the anterior insula would suggest that the improvers are perceiving the stimulus as less salient. This could be at- tributed to a number of reasons such as better mood, better coping skills learned from the CBT intervention, and better health outcomes overall. Future research would need to investigate the role of the interaction be- tween the salience and sensorimotor network to get a better understanding of these results. neuroimaging and multi-omics microbiome assessment to evaluate the BGM axis. Although the sample size of this study was sufficient to identify robust changes in symp- toms, brain parameters, and gut microbial abundances, it limited our ability to identify statistically significant differ- ences between responders and non-responders in micro- bial metabolites. Other limitations include the lack of a reference healthy control population, validation cohort, or long-term follow-up to evaluate the durability of the microbiome and neuroanatomical changes in CBT responders. Authors’ contributions JPJ, AG, JML, and EAM did the study concept and design. KT, VL, RF, GG, BME, JSL, and BDN took part in the acquisition of data. JPJ, AG, RRB, JB, KG, and VL are responsible for the analysis and interpretation of data. JPJ, AG, JML, and EAM drafted the manuscript. JPJ, AG, RRB, JB, KG, KT, VL, RF, GG, BME, JSL, BDN, JML, and EAM critically revised the manuscript for important intellectual content. JPJ and AG contributed in the statistical analysis. JML and EAM obtained funding. JML and EAM supervised the study. The authors read and approved the final manuscript. Conclusions Supplementary Information Conclusions This study demonstrates for the first time that a brief non-drug, non-dietary intervention that teaches informa- tion processing skills can modulate key components of the brain-gut-microbiome axis in IBS patients. More- over, their likelihood of treatment response could be predicted from baseline microbiota composition, raising the possibility that CBT-responsive IBS patients can be identified in clinical practice using microbial biomarkers. The observed changes in brain, gut microbes and symp- toms in patients who responded to this brain directed therapy supports the role of alterations in the brain-gut- microbiome axis in IBS, and is most consistent with an important influence of the brain on the gut microbiome. Larger studies are needed to characterize the functional correlates of gut microbial changes and to identify dis- tinct subtypes of IBS patients for whom brain- and gut- directed therapies are most effective. Treatment-induced changes were also seen in the di- versity and relative abundances of the gut microbiota. While there was a decrease in richness and phylogenetic diversity, five genera were enriched in responders, with Bacteroides being the most abundant. Thus, while low levels of Bacteroides abundance prior to treatment were predictive of a response, CBT increased the abundance of this genus, resulting in a conversion of many patients to Bacteroides-predominant microbiota. The reduction in microbial diversity in CBT responders may reflect dis- placement of enteric microbes by the expanded Bacter- oides population. As CBT only targets brain and behavioral mechanisms and there was no evidence of CBT-associated alterations in dietary intake in re- sponders, the change in microbiome is likely secondary to effects of CBT on the brain. Further supporting the role of the brain in these microbial abundance changes, the observed functional and structural brain changes were associated with an increase in Bacteroides and un- classified S24-7, a family of microbes belonging to the order of Bacteroidales. These findings suggest that a gut microbial community state (Clostridiales high, Bacter- oides low) is responsive to top-down signals from the brain affected by the cognitive skills learned through CBT, resulting in conversion to an alternate state enriched in Bacteroides. While the association of this microbial change with clinical improvement suggests its importance in symptom generation, baseline microbial composition was not significantly associated with sever- ity by IBS-SSS. This argues that microbial change alone is insufficient for protection against symptom generation in IBS and emphasizes the importance of brain alter- ations for CBT response. Funding P g JPJ was supported by VA CDA2 IK2CX001717. AG was supported by NIH grants K23 DK106528 and R03 DK121025. EAM was supported by NIH grants R01 DK048351 and R01 DK064539. JML was supported by R01 DK096606, U01 DK077738, Office of UB Vice Provost of Research and Genomic The strengths of this study are its prospective, longitu- dinal design and incorporation of both multimodal Page 13 of 14 Page 13 of 14 Jacobs et al. Microbiome (2021) 9:236 Jacobs et al. Microbiome Environment and Microbiome Research. This work was also supported by P30 DK041301 and P50 DK082370. 4. Francis CY, Morris J, Whorwell PJ. The irritable bowel severity scoring system: a simple method of monitoring irritable bowel syndrome and its progress. Aliment Pharmacol Ther. 1997;11(2):395–402. https://doi.org/10.104 6/j.1365-2036.1997.142318000.x. 4. Francis CY, Morris J, Whorwell PJ. The irritable bowel severity scoring system: a simple method of monitoring irritable bowel syndrome and its progress. Aliment Pharmacol Ther. 1997;11(2):395–402. https://doi.org/10.104 6/j.1365-2036.1997.142318000.x. Availability of data and materials 5. Spiegel B, Camilleri M, Bolus R, Andresen V, Chey WD, Fehnel S, Mangel A, Talley NJ, Whitehead WE: Psychometric evaluation of patient-reported outcomes in irritable bowel syndrome randomized controlled trials: a Rome Foundation report. Gastroenterology 2009, 137(6):1944-1953 e1941-1943. 5. Spiegel B, Camilleri M, Bolus R, Andresen V, Chey WD, Fehnel S, Mangel A, Talley NJ, Whitehead WE: Psychometric evaluation of patient-reported outcomes in irritable bowel syndrome randomized controlled trials: a Rome Foundation report. Gastroenterology 2009, 137(6):1944-1953 e1941-1943. The sequencing data supporting the conclusions of this article are available in the NCBI Bioproject repository, PRJNA736955 (https://www.ncbi.nlm.nih.gov/ bioproject/736955). The untargeted metabolomics data have been deposited in the Metabolomics Workbench repository under ID 2820 (https://www. metabolomicsworkbench.org). Short-chain fatty acid data are included with this article as a supplementary data file. The deidentified and raw neuroimaging data are available through the NIH funded PAIN (pain and interoception im- aging network) data repository (https://www.painrepository.org/repositories), which includes multimodal MRI scans in healthy controls and various chronic pain conditions. Access to the data is based on membership which is based on a collaborative principle. 6. Mayer EA, Labus JS, Tillisch K, Cole SW, Baldi P. Towards a systems view of IBS. Nat Rev Gastroenterol Hepatol. 2015;12(10):592–605. https://doi.org/10.1 038/nrgastro.2015.121. 6. Mayer EA, Labus JS, Tillisch K, Cole SW, Baldi P. Towards a systems view of IBS. Nat Rev Gastroenterol Hepatol. 2015;12(10):592–605. https://doi.org/10.1 038/nrgastro.2015.121. 7. Spohn SN, Mawe GM. Non-conventional features of peripheral serotonin signalling - the gut and beyond. Nat Rev Gastroenterol Hepatol. 2017;14(7): 412–20. https://doi.org/10.1038/nrgastro.2017.51. 8. Mayer EA, Labus J, Aziz Q, Tracey I, Kilpatrick L, Elsenbruch S, et al. Role of brain imaging in disorders of brain-gut interaction: a Rome Working Team Report. Gut. 2019;68(9):1701–15. https://doi.org/10.1136/gutjnl-2019-318308. 8. Mayer EA, Labus J, Aziz Q, Tracey I, Kilpatrick L, Elsenbruch S, et al. Role of brain imaging in disorders of brain-gut interaction: a Rome Working Team Report. Gut. 2019;68(9):1701–15. https://doi.org/10.1136/gutjnl-2019-318308. Ethics approval and consent to participate Institutional Review Boards at each of two sites approved the protocols governing participants: University at Buffalo (IRB 030-396754 and 030-745083) and Northwestern University (IRB STU00070073). All subjects provided in- formed consent to participate. 10. Labus JS, Osadchiy V, Hsiao EY, Tap J, Derrien M, Gupta A, et al. Evidence for an association of gut microbial Clostridia with brain functional connectivity and gastrointestinal sensorimotor function in patients with irritable bowel syndrome, based on tripartite network analysis. Microbiome. 2019;7(1):45. https://doi.org/10.1186/s40168-019-0656-z. 10. Labus JS, Osadchiy V, Hsiao EY, Tap J, Derrien M, Gupta A, et al. Evidence for an association of gut microbial Clostridia with brain functional connectivity and gastrointestinal sensorimotor function in patients with irritable bowel syndrome, based on tripartite network analysis. Microbiome. 2019;7(1):45. https://doi.org/10.1186/s40168-019-0656-z. Declarations 9. Simren M. Manipulating the gut microbiome as a treatment strategy for functional gastrointestinal disorders. Gastroenterology. 2018;155(4):960–2. https://doi.org/10.1053/j.gastro.2018.09.008. 9. Simren M. Manipulating the gut microbiome as a treatment strategy for functional gastrointestinal disorders. Gastroenterology. 2018;155(4):960–2. https://doi.org/10.1053/j.gastro.2018.09.008. Consent for publication Not applicable 11. Labus JS, Hollister EB, Jacobs J, Kirbach K, Oezguen N, Gupta A, et al. Differences in gut microbial composition correlate with regional brain volumes in irritable bowel syndrome. Microbiome. 2017;5(1):49. https://doi. org/10.1186/s40168-017-0260-z. 11. Labus JS, Hollister EB, Jacobs J, Kirbach K, Oezguen N, Gupta A, et al. Differences in gut microbial composition correlate with regional brain volumes in irritable bowel syndrome. Microbiome. 2017;5(1):49. https://doi. org/10.1186/s40168-017-0260-z. Author details 1G O h 1G. Oppenheimer Center for Neurobiology of Stress and Resilience, UCLA, Los Angeles, CA, USA. 2David Geffen School of Medicine, UCLA, Los Angeles, CA, USA. 3Vatche and Tamar Manoukian Division of Digestive Diseases, UCLA, Los Angeles, CA, USA. 4Division of Gastroenterology, Hepatology and Parenteral Nutrition, Veterans Administration Greater Los Angeles Healthcare System, Los Angeles, CA, USA. 5Imaging Genetics Center, Mark and Mary Stevens Institute for Neuroimaging and Informatics, Keck School of Medicine at USC, University of Southern California, Los Angeles, USA. 6Division of Behavioral Medicine, Jacobs School of Medicine, University at Buffalo, SUNY, Buffalo, NY, USA. 7Department of Radiological Sciences, UCLA, Los Angeles, CA, USA. 8Department of Psychiatry and Biobehavioral Sciences, UCLA, Los Angeles, CA, USA. 9G. Oppenheimer Center for Neurobiology of Stress and Resilience, Vatche and Tamar Manoukian Division of Digestive Diseases, 15. Martin CR, Osadchiy V, Kalani A, Mayer EA. 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Gastroenterology. 2006;130(5):1377–90. https://doi.org/10.1053/j.ga stro.2006.03.008. David School of Medicine at UCLA, CHS 42-210 MC737818, 10833 Le Conte Avenue, Los Angeles, USA. 18. Lackner JM, Jaccard J, Group IBSOSR. Factors associated with efficacy of cognitive behavior therapy vs education for patients with irritable bowel syndrome. Clin Gastroenterol Hepatol. 2019;17(8):1500–8 e1503. https://doi. org/10.1016/j.cgh.2018.10.033. 18. Lackner JM, Jaccard J, Group IBSOSR. Factors associated with efficacy of cognitive behavior therapy vs education for patients with irritable bowel syndrome. Clin Gastroenterol Hepatol. 2019;17(8):1500–8 e1503. https://doi. org/10.1016/j.cgh.2018.10.033. Received: 15 June 2021 Accepted: 4 November 2021 Received: 15 June 2021 Accepted: 4 November 2021 19. Lackner JM, Jaccard J, Radziwon CD, Firth RS, Gudleski GD, Hamilton F, et al. Durability and decay of treatment benefit of cognitive behavioral therapy for irritable bowel syndrome: 12-month follow-up. Am J Gastroenterol. 2019; 114(2):330–8. https://doi.org/10.1038/s41395-018-0396-x. 19. Lackner JM, Jaccard J, Radziwon CD, Firth RS, Gudleski GD, Hamilton F, et al. Competing interests 12. Van Oudenhove L, Crowell MD, Drossman DA, Halpert AD, Keefer L, Lackner JM, et al. Naliboff BD. Biopsychosocial aspects of functional gastrointestinal disorders. Gastroenterology: Levy RL; 2016. 12. Van Oudenhove L, Crowell MD, Drossman DA, Halpert AD, Keefer L, Lackner JM, et al. Naliboff BD. Biopsychosocial aspects of functional gastrointestinal disorders. Gastroenterology: Levy RL; 2016. EAM is a scientific advisory board member of Danone, Axial Biotherapeutics, Viome, Amare, Mahana Therapeutics, Pendulum, Bloom Biosciences, APC Microbiome Ireland. BME is an advisor for Hoffman La-Roche; Siemens; Nati- vis; Medicenna; MedQIA; Bristol Meyers Squibb; Imaging Endpoints; VBL; and Agios Pharmaceuticals. BME is a Paid Consultant for Nativis; MedQIA; Sie- mens; Hoffman La-Roche; Imaging Endpoints; Medicenna; and Agios. BME re- ceived grant funding from Siemens, Agios, and Janssen. JPJ, AG, RRB, JB, KG, KT, VL, RF, GG, JSL, BDN, and JML do not have any disclosures. 13. Ford AC, Lacy BE, Harris LA, Quigley EMM, Moayyedi P. Effect of antidepressants and psychological therapies in irritable bowel syndrome: an updated systematic review and meta-analysis. Am J Gastroenterol. 2019; 114(1):21–39. https://doi.org/10.1038/s41395-018-0222-5. 13. Ford AC, Lacy BE, Harris LA, Quigley EMM, Moayyedi P. Effect of antidepressants and psychological therapies in irritable bowel syndrome: an updated systematic review and meta-analysis. Am J Gastroenterol. 2019; 114(1):21–39. https://doi.org/10.1038/s41395-018-0222-5. 14. 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https://openalex.org/W4313206571	OpenAlex	Open Science	CC-By	2,022	A MATHEMATICAL MODEL FOR ACCESS CONTROL THROUGH TRUST MANAGEMENT IN UBIQUITOUS COMPUTING	D. S. SHELAR	English	Spoken	4,972	8,401	1. INTRODUCTION In day to day life, the role of computer technology is to integrate the public and private sectors fields such as trade, logistics, industry, transport, and healthcare through electronic devices. To address this issue, Mark Weiser [1] first time introduced the paradigm shift from one to one computer to one to many computers through ubiquitous computing (Ubicomp) in which electronic nodes are fully connected and make them available for sharing and processing the data because Ubicomp has capacity to remove the complexities of computing so that the various activities can be done efficiently with optimal use of memory. Ubiquitous computing is the paradigm which postulates a world where people are surrounded by computing devices and a computing infrastructure that supports each other in everything that people perform in the physical world. The focus is on developing ubiquitous computer systems to support people in their daily activities of various tasks of the physical world even though the electronic nodes are mobile. Ubicomp provides access to all electronic user nodes on the basis of smart interactions. The smart interaction refers to the interaction made with the credible nodes which provide trustworthy information. If a user is trustworthy, the experience will be pleasant even if the product is not perfect in a business transaction. If node A trusts node B and then they will remain loyal to each other because loyal nodes are incredibly useful in the strategy of access control to grant access to other electronics nodes. This Means, when we are able to interact in such a way that builds trust, we naturally build personal credibility. The emergence of these devices has created many security issues for which solutions and existing mechanisms are sufficient, especially concerning the problems of authentication and user’s private data protection. So, one of the important threats of ubiquitous computing is to maintain privacy which may cause its popularity to decrease. Many researchers contributed a lot to data security in access control for Ubicomp. Hua Wang [2] and his team presented an access control model to protect services and devices in the Ubicomp environment, which allows the access restrictions directly on services and object documents. This model provides a mechanism to build relationships between models and objects. Finally, comparisons with related works are analyzed. Several studies are also available on the security requirements for ubiquitous computing ([3]–[6]). A MATHEMATICAL MODEL FOR ACCESS CONTROL THROUGH TRUST MANAGEMENT IN UBIQUITOUS COMPUTING D. S. SHELAR, P. G. ANDHARE, S. B. GAIKWAD AND P. A. THAKRE D. S. SHELAR, P. G. ANDHARE, S. B. GAIKWAD AND P. A. THAKRE ABSTRACT In the ubiquitous computing environment of wireless access control, access is granted on having successful interactions with other devices/nodes/agents. Consequently, the wireless technologies further extend the association between devices to interact with each other with respect to provide access control. For mobile nodes, many times associations are volatile in nature. There is always a risk associated with such interactions when there is no trust amongst them. From a security perspective, there is a requirement for a proper mechanism to build trust so that nodes can interact with each other for the safe sharing of information. Many researchers have proposed trust models in which the credibility, mobility, and reliability of requester agents are not considered. In this study, we proposed the trust calculation model in ubiquitous computing by considering parameters like credibility, transitivity, reliability, and mobility. This trust calculation model can be incorporated in Ubiquitous Computing for better decision making for access control. Keywords and phrase: Access Control; Ubiquitous Computing; Trust; Credibility; Mobility; Reliability; Transitivity. Transitivity. Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Revised: 19th January 2022 Received: 5th January 2022 Accepted: 10th February 2022 2. MOTIVATION In electronic social media (ESM) anyone can freely share information on the basis of relationships through communication and even develop a new relationship with others who are strangers. But a relationship with strangers may cause to disclose privacy. So, it is necessary to check the trustworthiness of the people in the process of information sharing. Therefore there should be an access control policy regulated by the owner. Many traditional access control mechanisms are available for ESM ([19]-[21]). But it has its own limitations because the owner-user relationship is dynamic. After a certain period of time users may disclose the owner’s private information. So it becomes necessary to monitor the behavior of user continuously and check their trustworthiness of users. In order to address this issue, Baek and Kim [22] proposed a dynamic trust-based access control for the online social network in which they evaluated the dynamic trust values so that owners can monitor negative behavior and decide whether the access should be granted to a user or not. Since the level of access control from device A to device B is directly proportional to the trust device A is holding for device B [7]. Access control and trust are closely related as the level of access granted by a particular device to another device or service depend on the level of trust between these devices. Over the time trust has been an established concept in network security and therefore it is treated as an emerging field of ubiquitous computing with respect to access control. Many existing studies identified that the trust as an alternative to collaborate in a world without passwords and necessary certificates. It seems to be the logical choice that we are simply to integrate its workings and mechanisms into our new security concepts for ubiquitous computing. Even though trust- based access control is gaining momentum in the field of ubiquitous computing by allowing electronic agents to compute the ‘trustworthiness’ of other electronic agents based on their interactions, much remains unclear when it comes to defining the problem we are trying to solve. So, it is obvious to raise the question, what is the role of trust in access control for Ubicomp? This type of question can be addressed by giving access to the agent by the positive interactions happening with even third party users. 2. MOTIVATION In most of the work related to ubicomp, trust has focused on expanding the trust concepts from network security, i.e. granting or denying access not simply based on pre-computed certificates, but rather depending on a particular context. By taking inspiration from the earlier contribution regarding the role of trust for denial or granting the access in Ubicomp. We have been motivated to present the model for trust calculation by considering parameters like credibility, reliability, transitivity, and mobility. Special Issue on Recent Research on Management, Applied Sciences and Technology 1. INTRODUCTION In this study, we have proposed a mathematical model for the calculation of trust of an agent in access control for Ubicomp by considering the factors like credibility, reliability, transitivity, and mobility of an agent and tried to present the best solution for access control. The remaining section of the paper is organized as follows: Section 2 explains the motivation for our model. Section 3 explains the literature review of the various approaches regarding trust models in Ubicomp. The proposed mathematical model is presented in Section 4 followed by Section 5 and 6 which explain results and conclusion respectively. 1002 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Mahalle et al. [7] also presented a Fuzzy approach to the Trust Based Access Control model where the trust is calculated using uncertain parameters such as experience knowledge, recommendation. Nalini Mhetre and her co-workers [8] presented an experience model for Ubicomp. In which they proposed a mathematical model for experience of nodes with other nodes by considering the parameters such as history, reliability, and transitivity and ubiquity so as to grant access control. In this study, we have proposed a mathematical approach for access control in Ubicomp using the trustworthiness of agents. But while calculating trustworthiness in [[7]-[8]] the factors like credibility and mobility of an electronic agents are not taken into consideration, so we proposed a trust calculation model by considering those factors. 5. PROPOSED MODEL Existing fuzzy trust based access control (FTBACM) model calculate trustworthiness of each device or group of devices based on three components such as knowledge, experience and recommendation but the proposed trustworthiness model presented in this article with attributes like like Credibility (𝐶𝑟), Reliability (𝑅𝑙), Transitivity (𝑇𝑟) and Mobility (𝑀𝑏) of requester agent where 𝐶𝑟, 𝑅𝑙 and 𝑇𝑟 are the real numbers belongs to [0,1]. 4. CHALLENGES Many researchers presented an access control models using trust in ubiquitous computing but those models are not fully acceptable due to privacy issues of owners and prevention of malicious users. One of the reasons for creating this issue is to have inappropriate trust value of user-node by considering limited attributes such as recommendations (by the third party), history, transitivity, and reliability and also the calculation of each attribute. In proposed the model we have considered credibility and mobility of user-node so that on the basis of previous interactions we can calculate up to how much degree is the user-node node credible? 5.1 Credibility Recently electronic social media has become an integrated part of everyday life. However, the owners are facing challenges of being susceptible to wrong information and rumors. Therefore the credibility of user nodes in social media needs to be checked. Castillo et al. [24] presented in his seminal paper on information credibility of news propagated through Twitter using user’s profile information, network information, and user’s behavior (tweets and retweets) to assess the credibility of tweets. Barbier and Liu [23] proposed a method to find provenance paths leading to sources of the information to evaluate its credibility. A research article by Jiaxi Sun [25], evaluated the credibility of social media information based on user perception by considering three parameters such as subject credibility, source credibility and content credibility of electronic social media. Looking toward the significance of the credibility of user nodes, we considered this attribute in order to increase the effectiveness of trust value. Credibility component of any device can be calculated on the basis of past interaction that happened with other devices. Rating of the past interaction is done to be 0 to 1. If the interaction factor is close to 1 then it is said to be successful and if it is close to zero then it is said to be unsuccessful. Let 𝐼𝑗 be the past interaction happened where 𝐼𝑗∈[0,1], 𝑗= 1,2, ⋯, 𝑛. Credibility (𝐶𝑟) = [(∑ 𝑗=𝑛 𝑗=1 𝐼𝑗)/𝑛] and the values of 𝐶𝑟∈[0,1]. Credibility (𝐶𝑟) = [(∑ 𝑗=𝑛 𝑗=1 𝐼𝑗)/𝑛] and the values of 𝐶𝑟∈[0,1]. Credibility (𝐶𝑟) = [(∑ 𝑗=𝑛 𝑗=1 𝐼𝑗)/𝑛] and the values of 𝐶𝑟∈[0,1]. If 𝐶𝑟≥0.8 then the agent is treated as credible(to be decided by owner-agent) . If 𝐶𝑟≥0.8 then the agent is treated as credible(to be decided by owner-agent) . 3. RELATED LITERATURE In ubiquitous computing, the trust amongst the agents is one of the important factor for rendering access to other agents. It also helps to deal with security issues. Many researchers have presented their views through their work. Matt Blaze et al. [9] first proposed decentralized trust-management Policymaker where they have presented the comprehensive approach to the trust management, based on a simple language for specifying trusted actions and trust relationships that will facilitate the development of security features in a wide range of network services. Jameel et al.[10] described the trust model which is based on the vectors of trust values of different entities in ubiquitous computing by capturing uncertainties occurred in his model. The trust value is calculated by considering various factors such as peer reputation, confidence, history of past interaction, and time-based trust evaluation factor. Also efficiently handled the false recommendations of entities. Colin English [11] and his team has intended to assume that the trust and cryptographic security are considered to be orthogonal to each other by assuming the existence of reliable encryption techniques and focus on the characteristics of a model that supports the management of the trust relationships between two devices during ad-hoc interactions. R He et al. [12] developed a trust management framework for a multi-cloud environment to effectively evaluate the trustworthiness of Cloud Service Providers and Trust Service Providers based trust management architecture using subjective and objective trust. Prajapati et al. [14] introduced a trust management model to manage the trust and its properties for software as a service in a cloud computing environment. The Trust model is constructed based on several trust properties such as asymmetry, reflexivity, context dependency, scalability, partial transitivity and Uncertainty. Malika Yaici et al.[15] proposed context aware authentication system using trust management between client and servers even among servers. Also presented is an algorithm to calculate context based trust. 1003 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) Since the magnitude of trust between agents A, B and B, C is not the same. So in order to minimize the risk, we considered the trust between A and C is the magnitude of the minimum of trust between A, B and B, C. Let 𝛼, 𝛽, 𝛾 be the trust between (A, B), (B, C) and (A, C). Let 𝛼, 𝛽, 𝛾 be the trust between (A, B), (B, C) and (A, C). Let 𝛼, 𝛽, 𝛾 be the trust between (A, B), (B, C) and (A, C). 𝛾= 𝑚𝑖𝑛 (𝛼, 𝛽) 𝛾= 𝑚𝑖𝑛 (𝛼, 𝛽) 𝛾= 𝑚𝑖𝑛 (𝛼, 𝛽) 5.4 Mobility Mobility is one of the important aspects when a large number of agents play in Ubicomp. In online social networks also users provide access to unknown requester-agents even if it is mobile (not stationary) to build relationships. If requester-agents move far away from user-agent, in this case the proper interaction may not happen which may result in the risk of disseminating unclear information. In order to address this issue traditional role based access control models are applied in online social networking. But those models are not suitable for the non-stationary environment because user behaviour is unpredictable in terms of distance. So, we used the mobility factor range of requester-agents which is one of the important contributors for trust calculations. The remarkable permission to the agent is granted when the distance of the requester-node from the user-node is within the specified range. Let 𝑟 and 𝑜 be the positions of requester-agent and user-agent respectively. tions of requester-agent and user-agent respectively. Let 𝑟 and 𝑜 be the positions of requester-agent and user-agent respectively. Let s be the distance between 𝑟 and 𝑜 that is 𝑑(𝑟, 𝑜) = 𝑠 Let s be the distance between 𝑟 and 𝑜 that is 𝑑(𝑟, 𝑜) = 𝑠 and 𝑅(𝑟,𝑜) be the region covered by user-agent. and 𝑅(𝑟,𝑜) be the region covered by user-agent. 𝑅(𝑟,𝑜) = {𝑥/𝑑(𝑟𝑖, 𝑜𝑖) ≤𝑠} 𝑅(𝑟,𝑜) = {𝑥/𝑑(𝑟𝑖, 𝑜𝑖) ≤𝑠} If a requester-agent lies in the region 𝑅(𝑟,𝑜) then the interaction may happen whether it is successful or nor it is immaterial. In this case we assign the value of mobility factor is 1 else 0. 5.3 Reliability y Trust is incomplete without accurate reliability. In the Ubicomp environment the access control, the reliability of an agent to which the access is granted is highly needed to be known. The reliability of an agent can be calculated on the basis of the degree to which the agent shows consistently good performance in interactions. Node A is reliable to node B only if A consistently keeps good interactions with B. Consistently good interactions in a short period of time or with less number of good consistent interactions may invite less reliability. This degeneracy can be overcome by finding the coefficient of correlation between two sets of interaction values. Reliability of an agent is one of the important parameter in wireless sensor network (WSN). Access is granted when an agent is highly reliable. There is a set of interactions that happened between two agents in two time periods in months or years. If two sets of interactions are correlated up to the desired level then the agent is reliable. Let (𝐼𝑗)𝑡1 and (𝐼𝑗)𝑡2 be the interaction values between two agents in time slot 𝑡1 and 𝑡2 respectively. The correlation coefficient between (𝐼𝑗)𝑡1 and (𝐼𝑗)𝑡2 is 𝑟𝑙= ∑ 𝑗=𝑛 𝑗=1 {[(𝐼𝑗)𝑡1−(̅𝐼𝑗)𝑡1][(𝐼𝑗)𝑡2−(̅𝐼𝑗)𝑡2]} √∑ 𝑗=𝑛 𝑗=1 [(𝐼𝑗)𝑡1−(̅𝐼𝑗)𝑡1] 2 ∑ 𝑗=𝑛 𝑗=1 [(𝐼𝑗)𝑡2−(̅𝐼𝑗)𝑡2] 2 (1) 𝑟𝑙= ∑ 𝑗=𝑛 𝑗=1 {[(𝐼𝑗)𝑡1−(̅𝐼𝑗)𝑡1][(𝐼𝑗)𝑡2−(̅𝐼𝑗)𝑡2]} √∑ 𝑗=𝑛 𝑗=1 [(𝐼𝑗)𝑡1−(̅𝐼𝑗)𝑡1] 2 ∑ 𝑗=𝑛 𝑗=1 [(𝐼𝑗)𝑡2−(̅𝐼𝑗)𝑡2] 2 (1) where (1) where 1. (̅𝐼𝑗)𝑡1 and (̅𝐼𝑗)𝑡2 be the mean values of (𝐼𝑗)𝑡1 and (𝐼𝑗)𝑡2. 1. (̅𝐼𝑗)𝑡1 and (̅𝐼𝑗)𝑡2 be the mean values of (𝐼𝑗)𝑡1 and (𝐼𝑗)𝑡2. 1. (̅𝐼𝑗)𝑡1 and (̅𝐼𝑗)𝑡2 be the mean values of (𝐼𝑗)𝑡1 and (𝐼𝑗)𝑡2. 2. −1 ≤𝑟≤1. For convenience the reliability factor we considered 𝑅𝑖= \|𝑟𝑙\|. 5.2 Transitivity y In trust management, an agent A is not directly interacted with an agent C but authentication is required between them where the trust transfer plays an important role through intermediate agent B. Trust transfer happens if an agent B recommend agent A that an agent C is trustworthy if an agent A is trustor and agent B is trustee. But the magnitude of trust between agents B and C and A and C may differ. Nowadays, the trustworthiness among the agents that do not know each other. In access control this is a big challenge. This issue may be addressed through trust transitivity. Many researchers addressed this issue, Josang and Pope [16] used belief theory for deriving transitive trust using the belief operators of subjective logic described trust transitivity in online interaction among strangers. Xiang Qiuet et al.[17] described a trust transitivity model based on evidence theory by capturing uncertainty occurring in the model. Xu and Fung [18] proposed a risk-defined trust transitivity model for group decision making in social networks with the help of a risk-defined trust propagation operator that can propagate trust and distrust information based on risk bearing level defining factors like trust, distrust, uncertainty and inconsistency. 1004 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL 5.5 Trust (T) Calculation 5.5 Trust (T) Calculation Step1:If the mobility factor of the requester agent is 1 then calculate trust of a requester-agent using the following formula. the mobility factor of the requester agent is 1 then calculate trust of a requester-agent usi formula. (2) (2) 𝑇= 𝑤1𝐶𝑟+ 𝑤2𝑇𝑟+ 𝑤3𝑅𝑙 1005 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL where 𝑇, 𝑤1, 𝑤2, 𝑤3 ∈[0,1] suchthat ∑𝑤𝑘 𝑘=3 𝑘=1 = 1 where 𝑇, 𝑤1, 𝑤2, 𝑤3 ∈[0,1] suchthat ∑𝑤𝑘 𝑘=3 𝑘=1 = 1 Also 𝑤1, 𝑤2 and 𝑤3 are the weights assigned to credibility, transitivity and reliability respectively. If the 𝑇 is close to 1, the agent/requester node is said to be trustworthy. Step:2If the mobility factor of the requester agent is 0 then ignore the requester-agent that is access should be denied. Figure 1: Flow Chart 5.6 Experimental Results An experiment on 15 user/requester nodes is performed as per the significance of the attributes in 8 scenari We came to the conclusion that the trust value of an agent is as appropriate for the 60% weightage of reliabili 30% weightage of credibility and 10% weightage of transitivity. It means that the reliability factor is mo mportant in trust calculation. Figure 2 to Figure 9 shows the result of the trust calculation in this scenar However, the above weightage may vary from problem to problem and situation to situation. 5.6 Experimental Results 5.6 Experimental Results 5.6 Experimental Results An experiment on 15 user/requester nodes is performed as per the significance of the attributes in 8 scenarios. We came to the conclusion that the trust value of an agent is as appropriate for the 60% weightage of reliability, 30% weightage of credibility and 10% weightage of transitivity. It means that the reliability factor is more important in trust calculation. Figure 2 to Figure 9 shows the result of the trust calculation in this scenario. However, the above weightage may vary from problem to problem and situation to situation. p An experiment on 15 user/requester nodes is performed as per the significance of the attributes in 8 scenarios. We came to the conclusion that the trust value of an agent is as appropriate for the 60% weightage of reliability, 30% weightage of credibility and 10% weightage of transitivity. It means that the reliability factor is more important in trust calculation. Figure 2 to Figure 9 shows the result of the trust calculation in this scenario. However, the above weightage may vary from problem to problem and situation to situation. 1006 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL ol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) 1 i l I R R h M A li d S i d T h l UGC CARE APPROVED JOURN Figures : 2-9 CONCLUSION ubiquitous computing, simultaneous interactions within many to many nodes happen. In such vironment, malicious nodes may provide insufficient or incorrect information that may cause issue vacy and security. In order to address this issue, we presented a mathematical model for access co ough trust management in ubiquitous computing using parameters such as credibility, transitivity, reliab d mobility, which is the best suited to provide access to the requester-agent/node. Also shown that esented trust calculation leads to better decision making with respect to weightage of three attributes edibility, transitivity and reliability. es : 2-9 g REFERENCES REFERENCES 1. Weiser M., "The computer for the 21 century", ACM SIGMOBILE Mobile Computing and Communications Review, 3(3),1999, pp. 3–11. 1. Weiser M., "The computer for the 21 century", ACM SIGMOBILE Mobile Computing and Communications Review, 3(3),1999, pp. 3–11. 2. Hua Wang, Yanchun Zhang, Jinli Cao, "Access control management for ubiquitous computing", Future Generation Computer Systems,Vol. 24,2008,pp.870–878. 3. J. Seigneur, C. Jensen, "Trust enhanced ubiquitous payment without too much privacy loss", SAC ’04: Proceedings of the ACM Symposium on Applied Computing, ACM Press, New York, NY, USA, 2004, pp.1593-1599. 4. G. Sampemane, P. Naldurg, R. Campbell, "Access control for active spaces", ACSAC’ 02 Proceedings of the 18th Annual Computer Security Applications Conference, IEEE Computer Society, Washington, DC, USA, 2002, pp. 343-352. 5. P. Viswanathan, B. Gill, R. Campbell, "Security architecture in gaia", Tech. Rep., Champaign, IL, USA, (2001),pp.10-20. 6. J. Jai-muhtadi, R. Campbell, A. Kapadia, M. Mickunas, S. Yi, "Routing through the mist: Privacy preserving communication in ubiquitous computing environments", Proceedings of the 22nd International Conference on Distributed Computing Systems, ICDCS’02, IEEE Computer Society, Washington, DC, USA , 2002, pp.1-10 7. P. N Mahalle, P. A. Thakre, N.R. Prasad, R.Prasad, "A fuzzy approach to trust based access control in internet of things", 3rd International Conference on Wireless Communications, Vehicular Technology, Information Theory and Aerospace and Electronic Systems, VITAE, 2013, pp. 2–6. 8. N. Mhetre, A. Deshpande, P. Mahalle, and P. Thakre, "Experience Modelling For Ubiquitous Computing A Mathematical Approach", Turkish Journal of Computer and Mathematics Education, Vol.12 No.6,2021, pp.5476-5488.. 9. M. Blaze, J Feigenbaum, J Lacy, " Decentralized trust management", 17th IEEE Symposium on Security and Privacy, Oakland , 1996, pp. 164–173. 10. H. Jameel, LX Hung, U Kalim, A Sajjad, S Lee, Yk Lee, "A trust model for ubiquitous systems based on vectors of trust values", 7th IEEE International Symposium on Multimedia, Irvine, USA, Dec 2005,pp. 674-679. 11. C English, P Nixon, S Terzis, A McGettrick, H Lowe, "Security models for trusting network appliances", 5th IEEE International Workshop on Networked Appliances, Liverpool, Oct 2002, pp. 39–44 12. R. He, J Niu, M Yuan, J Hu , "A novel cloud-based trust model for pervasive computing", 4th International Conference on Computer and Information Technology, China, Sept 2004, pp. 693–700. 13. V Cahill, E Gray, J Seigneur, C Jensen, Y Chen, B Shand, N Dimmock, A. 6. CONCLUSION In ubiquitous computing, simultaneous interactions within many to many nodes happen. In such an environment, malicious nodes may provide insufficient or incorrect information that may cause issues of privacy and security. In order to address this issue, we presented a mathematical model for access control through trust management in ubiquitous computing using parameters such as credibility, transitivity, reliability and mobility, which is the best suited to provide access to the requester-agent/node. Also shown that the presented trust calculation leads to better decision making with respect to weightage of three attributes like credibility, transitivity and reliability. In ubiquitous computing, simultaneous interactions within many to many nodes happen. In such an environment, malicious nodes may provide insufficient or incorrect information that may cause issues of privacy and security. In order to address this issue, we presented a mathematical model for access control through trust management in ubiquitous computing using parameters such as credibility, transitivity, reliability and mobility, which is the best suited to provide access to the requester-agent/node. Also shown that the presented trust calculation leads to better decision making with respect to weightage of three attributes like credibility, transitivity and reliability. In ubiquitous computing, simultaneous interactions within many to many nodes happen. In such an environment, malicious nodes may provide insufficient or incorrect information that may cause issues of privacy and security. In order to address this issue, we presented a mathematical model for access control through trust management in ubiquitous computing using parameters such as credibility, transitivity, reliability and mobility, which is the best suited to provide access to the requester-agent/node. Also shown that the presented trust calculation leads to better decision making with respect to weightage of three attributes like credibility, transitivity and reliability. 1007 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL 18. Jun Xu, Carol J. Fung, "dblp Computer Science Biblioraphy", A Risk-defined Trust Transitivity Model for Group Decisions in Social Networks,(2019), pp.415-420. 19. X. Qian, T. F. Lunt, "A MAC policy framework for multilevel relational databases", IEEE Transactions on Knowledge and Data Engineering, vol. 8, no. 1, 1996, pp.3–15 20. L. Snyder, Formal models of capability-based protection systems", IEEE Transactions on Computers, vol. 30, no. 3, 1981, pp. 172– 181. 21. R. S. Sandhu, E. J. Coyne, H. L. Feinstein, and C. E. Youman, "Role-based access control models", Computer, vol. 29, no. 2,1996, pp. 38–47. 22. Seungsoo Baek, Seungjoo Kim , "Trust-Based Access Control Model from Sociological Approach in Dynamic Online Social Network Environment", The Scientific World Journal, vol.1, 2014, pp.1-8 23. G. Barbier and H. Liu, "Information provenance in social media. Social Computing", Behavioral-Cultural Modeling and Prediction, 2011, pp. 276–283. 24. C. Castillo, M. Mendoza, and B. Poblete, "Information credibility on twitter", In Proceedings of the 20th international conference on World wide web,ACM, 2011, pp. 675–684. 25. Jiaxi Sun, “Research on the Credibility of Social Media Information Based on User Perception”, Security and Communication Networks Volume 2021,pp.1-10. AUTHOR DETAILS: 1D. S. SHELAR, 2P. G. ANDHARE, 3S. B. GAIKWAD AND 4P. A. THAKRE 1Department Engineering Sciences, AISSMS Institute of Information, Technology, Pune, India 2Department of Mathematics, RBNB College of Art, Commerce and Science, Srirampur, India 3Department of Mathematics, New Art, Commerce and Science College, Ahmednagar, India 4Department of Engineering Sciences, Zeal College of Engineering and Research, Pune, India REFERENCES Twigg, J Bacon, W Wagealla, S Terzis, P Nixon, G Serugendo, C Bryce, M Carbone, K Krukow, M Nielsen, "Using trust for secure collaboration in uncertain environments", IEEE Pervasive Computing Mobile and Ubiquitous Computing, 2,2003, pp. 52-61 14. S. Prajapati, S. Changder, A. Sarkar, "Trust Management Model for Cloud Computing Environment", Proceding of the International Conference on Computing. Communication and Advanced Network - ICCCAN, 2013, pp.509-513. 15. Malika Yaici, A. Oussayah, M. Ahmed. Takerrabet, "Trust Management for an Authentication System in Ubiquitous Computing", International Journal of Electronics and Communication Engineering , Vol:12, No:6, 2018 pp.469-478. 16. Jøsang Simon, Siman Pope, "Semantic, Constraints for Trust Transitivity, Conceptual Modelling", Second Asia-Pacific Conference on Conceptual Modelling (APCCM2005), Newcastle, NSW, Australia, 2005, pp. 115-124 17. Xiang Qiuet, Li Zhang, Shouxin Wang and Guanqun Qian, "A Trust Transitivity Model Based-on Dempster-Shafer Theory", Journal of Networks, Vol. 5, No. 9, 2010, pp.1025-1032. 1008 Special Issue on Recent Research on Management, Applied Sciences and Technology Stochastic Modeling & Applications Vol. 26 No. 3 (January - June, Special Issue 2022 Part - 2) ISSN: 0972-3641 UGC CARE APPROVED JOURNAL AUTHOR DETAILS: 1D. S. SHELAR, 2P. G. ANDHARE, 3S. B. GAIKWAD AND 4P. A. THAKRE 1Department Engineering Sciences, AISSMS Institute of Information, Technology, Pune, India 2Department of Mathematics, RBNB College of Art, Commerce and Science, Srirampur, India 3Department of Mathematics, New Art, Commerce and Science College, Ahmednagar, India 4Department of Engineering Sciences, Zeal College of Engineering and Research, Pune, India 1009 Special Issue on Recent Research on Management, Applied Sciences and Technology	41,783
W3117929855.txt_1	German-Science-Pile	Open Science	Various open science	null	None	None	German	Spoken	373	808	Hallers Krankheit in den Vordergrund; Pringle ist es, der dem schmerzgeplagten Freund zum Gebrauch von Opiumklistieren rät und auch nicht zögert, die Steigerung der Dosis zu empfehlen. Wohl nur mit dieser palliativen Behandlung wird Hallers ausserordentliche Arbeitsleistung in den letzten vier Lebensjahren überhaupt möglich. Der Titel von Otto Sonntags mit grosser Sorgfalt und Präzision erarbeiteten, in Einleitung, Erläuterungen und Textwiedergabe mustergültigen Edition berücksichtigt die Besonderheiten der Quellenlage. Während sich über 80 Briefe von Pringle erhalten haben, sind Hallers Antworten nur indirekt zugänglich, und zwar lediglich durch unterschiedlich lange Zitate, die Pringle in seine thematisch geordneten Materialsammlungen aufgenommen hat; auf diese Weise sind 53 Briefe Hallers fassbar. Pringles Kollektaneen gelangten durch seine letztwillige Verfügung in den Besitz des «Royal College of Physicians» von Edinburgh mit der ausdrücklichen Auflage, dass sie niemals publiziert werden dürfen. Gemäss diesem Vermächtnis hat das «College» dem Herausgeber zwar die Einsichtnahme in die Texte, jedoch nur die Publikation von kurzen Zusammenfassungen erlaubt. Damit fehlen authentische Briefe Hallers in englischer Sprache, die dieser nach dem Urteil Pringles erstaunlich gut beherrschte. Urs Böschung, Bern Soukup, Rudolph Werner; Mayer, Helmut: Alchemistisches Gold, Paracelsistische Pharmaka. Laboratoriumstechnik im 16. Jahrhundert. Chemiegeschichtliche und archäometrische Untersuchungen am Inventar des Laboratoriums von Oberstockstall/Kirchberg am Wagram. Wien etc., Böhlau, 1997. X, 336 S. III. (Perspektiven der Wissenschaftsgeschichte, 10). öS 498.-; DM 69.80; SFr. 63.50. ISBN 3-205-98767-5. Grundlage für die vorliegende Untersuchung bilden die 1980 im Schloss Oberstockstall, Niederösterreich, in einer Grube entdeckten Überreste eines alchemischen Laboratoriums aus dem 16. Jahrhundert (vgl. auch oben S. 280). Anhand archäometrischer Untersuchungen gelang es. aus den an den Scherben haftenden chemisehen Rückständen auf die im Labor durchgeführten Operationen zurückzuschliessen. Der Vergleich mit den schriftlichen Quellen der praktischen Alchemie der Frühen Neuzeit erlaubte die Rekonstruktion einiger im damaligen Laboratorium vermutlich durchgeführter alchemischen Prozesse, so aus dem Probierwesen, der versuchten Transmutationsalchemie und der Chemiatrie. Der Leser erhält anhand historischer Erläuterungen, der Archäometrie der Objekte und den entsprechenden Passagen aus der zeitgenössischen Literatur eine gründliche Einführung in die damais üblichen alchemischen Verfahren, die Chemikalien und in einige chemiatrische Pharmaka. Im Anschluss findet sich ein ausführliches Literaturverzeichnis mit der einschlägigen Primär- und Sekundärliteratur. Die Publikation liefert einen wertvollen Beitrag zum Verständnis der frühneuzeitlichen Alchemie. Urs Leo Gantenbein, Winterthur 289 Downloaded from Brill.com 07/13/2024 12:04:10PM via free access.	16,748
https://openalex.org/W2774915114	OpenAlex	Open Science	CC-By	2,017	The Use of Poly-d,l-lactic Acid (PDLLA) Devices for Bone Augmentation Techniques: A Systematic Review	Marco Annunziata	English	Spoken	9,425	16,337	The Use of Poly-D,L-lactic Acid (PDLLA) Devices for Bone Augmentation Techniques: Marco Annunziata * ID , Livia Nastri, Gennaro Cecoro and Luigi Guida Marco Annunziata * ID , Livia Nastri, Gennaro Cecoro and Luigi Guida Multidisciplinary Department of Medical-Surgical and Dental Specialties, University of Campania “Luigi Vanvitelli”, Via Luigi De Crecchio, 6, 80138 Naples, Italy; livia.nastri@unicampania.it (L.N.); gennarocecoro@gmail.com (G.C.); luigi.guida@unicampania.it (L.G.) * Correspondence: marco.annunziata@unicampania.it; Tel.: +39-081-566-5515 Received: 8 November 2017; Accepted: 8 December 2017; Published: 13 December 2017 Received: 8 November 2017; Accepted: 8 December 2017; Published: 13 December 2017 Abstract: Poly-D,L-lactic acid (PDLLA) has been proposed in dentistry for regenerative procedures in the form of membranes, screws, and pins. The aim of this review was to evaluate the efﬁcacy of bone augmentation techniques using PDLLA devices. A literature search was carried out by two independent and calibrated reviewers. All interventional and observational studies assessing the efﬁcacy of bone augmentation techniques using PDLLA devices were included. Six studies were included. The relevant variability of design and methods impeded any qualitative or quantitative comparison. Ease of handling, absence of a re-entry phase, moldability of foils, and good soft-tissue response were appreciated characteristics of PDLLA devices. Some drawbacks such as the risk of membrane exposition, a prolonged adsorbability, and a tendency to a ﬁbrous encapsulation of the PDLLA devices have been described, although the clinical signiﬁcance of these ﬁndings is unclear. Clinical data about PDLLA devices for bone regeneration are very scarce and heterogenous. Well-designed randomized controlled trials comparing the use of PDLLA foils and pins with conventional membranes for bone regeneration are strongly encouraged in order to understand the real clinical beneﬁts/drawbacks of this technique. Keywords: poly-D,L-lactic acid; bone augmentation; systematic review molecules molecules Molecules 2017, 22, 2214; doi:10.3390/molecules22122214 www.mdpi.com/journal/molecules molecules molecules 1. Introduction Dental rehabilitation of partially or totally edentulous patients with oral implants has become a routine treatment modality in recent decades, with reliable long-term results [1–5], radically changing the ﬁeld of dentistry and improving patients’ quality of life. However, unfavorable local atrophic conditions of the alveolar ridge, due to agenesis, periodontal disease, neoplasia, or traumatic events, may impair implant placement, and potentially compromise the long-term survival of implant rehabilitations from a functional and esthetic viewpoint. In order to correct the unfavorable anatomy of atrophic sites, a number of alveolar ridge augmentation techniques have been developed, including autologous bone blocks from intra- and extra-oral donor sites, ridge splitting and expansion, distraction osteogenesis, sandwich osteoplasty, and guided bone regeneration (GBR) [6–8]. The use of membranes to promote bone regeneration following the principles of the guided bone regeneration (GBR) has become a standard of care in dentistry to correct bone defects at sites scheduled for implant placement, and the efﬁcacy of such a procedure has been largely conﬁrmed by long-term clinical results [9,10]. First membranes used for GBR application were made of expanded polytetraﬂuoroethylene (ePTFE), reinforced or not by titanium inserts (to increase their plasticity and dimensional stability), and provided excellent treatment results [11,12]. The ePTFE membranes, www.mdpi.com/journal/molecules 2 of 14 Molecules 2017, 22, 2214 however, need to be removed in a second surgical stage. Furthermore, their efﬁcacy is highly correlated to the wound healing course, being heavily jeopardized in case of wound dehiscence with consequent membrane exposure and infection [13,14]. To overcome such drawbacks, resorbable membranes made of animal-derived collagen or synthetic aliphatic polyesters were introduced, [15–17] with evident advantages in terms of simpliﬁed technique, no need of a second surgical stage and decreased patient morbidity. Nevertheless, in some cases, such membranes showed to be less effective than the non-resorbable ones, due to their reduced mechanical properties and a decreased barrier function, as a result of the rapid enzymatic degradation by macrophages and polymorphonuclear leukocytes [18]. p y g y p g p y p y Polylactic acid (PLA) is a semi-crystalline polymer with molecular weights of 180,000 to 530,000, a melting point of about 174 ◦C, and glass transition temperature of 57 ◦C. Depending on the L and D conﬁguration, it can exist in several distinct forms, such as poly-L-lactide (PLLA) and poly-D-lactide (PDLA), and it is also degraded via hydrolysis [19]. 1. Introduction By adding D-isomers into an L-isomer based polymerization system of PLA, polymer chains widen and cannot be packed as tightly as PLLA polymer chains. The resulting material, the poly-D,L-lactic acid (PDLLA), is characterized by biomechanical, thermal, rheological, and biological properties which may be modulated on the basis of the different proportions of L- and D-isomers composing the different PDLLA formulations. D-isomer, indeed, is characterized by a more rapid resorption and a less crystalline structure, whereas the L-isomer is characterized by a higher crystallinity and a less rapid resorption. Regarding the rheological properties of the material, the shear viscosity of the polymer increases with increasing L-isomer in the L/D-isomer mixture because of the increasing crystallinity of PLA [20]. Also, the glass transition temperature increase with increasing the amount of the L-isomer [21]. This material, like other resorbable materials, degrades and resorbs in two phases [22]. During the ﬁrst phase, water molecules hydrolyze the long polymer chains into shorter fragments. The second phase consists of a physiologic response of the body in which macrophages phagocyte and metabolize the short fragments which subsequently enter the citric acid cycle [23–25], and are transformed into carbon dioxide and water, subsequently excreted from the body, mainly through respiration. PDLLA has been proposed in dentistry for regenerative procedures in form of ﬂuid membranes and solid foils, screws, and pins. PDLLA ﬂuid membranes (Atrisorb, Atrix Labs, Fort Collins, CO, USA) mainly used for guided tissue regeneration (GTR), in some case for GBR, transforms to a solid barrier when contacted with water or aqueous solutions and solidiﬁes in situ. This ﬂuid membrane is composed of 37% of a liquid polymer of lactic acid that is dissolved in 63% N-methyl-2-pyrrolidone; polymer drops are mixed with saline drops in a ratio of one drop of saline every two drops of polymer; then a rapid mixing of polymer and saline is performed in order to drive off the N-methyl-2-pyrrolidone carrier. The solid membrane, pins, and screws (Resorb-X, KLS Martin, Tuttlingen, Germany), mostly used for GBR, are composed of a 50/50 mixture of D and L-isomers of PLA and is characterized by an extended resorption time and increased mechanical properties [26,27]. A number of preclinical studies have investigated the regenerative features [28], the biological properties [29–31] as well as the physical–mechanical properties [32–34] of these PDLLA devices. 1. Introduction On the contrary, very few studies have evaluated the clinical performance of PDLLA devices for bone regeneration, so that a comprehensive understanding of their potential role in this ﬁeld is still lacking. The aim of this review was to evaluate the efﬁcacy of bone augmentation techniques using PDLLA devices, through a systematic search of the current available scientiﬁc literature. Molecules 2017, 22, 2214 Systematic Search Strategy Search string: (pdlla OR “poly-D,L-lactic acid” OR “poly(L,D-lactide)” OR “poly-D-L-lactide” OR “poly-D,L-lactide” OR “poly(L/D-lactide)” OR “poly(L,D-lactic acid)” OR “poly-D,L-lactide acid” OR “Atrisorb” OR “Resorb-X” OR “SonicWeld” OR “SonicPins”) AND (oral OR dental OR bone) AND (membrane OR shell OR pin OR barrier OR screw OR mesh) Filter: Humans Language: No restrictions Electronic databases: MEDLINE/PubMed, The Cochrane Central Register of Controlled Trials (CENTRAL), ClinicalTrials.gov Inclusion criteria: - Study design: interventional and observational studies - Population: partially edentulous patients - No restriction on age or number of patients - Healthy individuals (no systemic diseases, no medications affecting platelet and bone functions) - Intervention: bone augmentation using PDLLA devices - Comparison (if appliable): any other augmentation technique or material - Outcome: qualitative and/or quantitative changes of hard tissue at the augmented sites. - Follow up duration: any Exclusion criteria: - Studies regarding osteosynthesis techniques or periodontal regeneration - Conference abstracts, pre-clinical studies, technical reports, and reviews. Systematic Search Strategy Search string: (pdlla OR “poly-D,L-lactic acid” OR “poly(L,D-lactide)” OR “poly-D-L-lactide” OR “poly-D,L-lactide” OR “poly(L/D-lactide)” OR “poly(L,D-lactic acid)” OR “poly-D,L-lactide acid” OR “Atrisorb” OR “Resorb-X” OR “SonicWeld” OR “SonicPins”) AND (oral OR dental OR bone) AND (membrane OR shell OR pin OR barrier OR screw OR mesh) Filter: Humans Language: No restrictions Electronic databases: MEDLINE/PubMed, The Cochrane Central Register of Controlled Trials (CENTRAL), ClinicalTrials.gov Inclusion criteria: - Study design: interventional and observational studies - Population: partially edentulous patients - No restriction on age or number of patients - Healthy individuals (no systemic diseases, no medications affecting platelet and bone functions) - Intervention: bone augmentation using PDLLA devices - Comparison (if appliable): any other augmentation technique or material - Outcome: qualitative and/or quantitative changes of hard tissue at the augmented sites. - Follow up duration: any Exclusion criteria: - Studies regarding osteosynthesis techniques or periodontal regeneration - Conference abstracts, pre-clinical studies, technical reports, and reviews. - Studies regarding osteosynthesis techniques or periodontal regeneration - Conference abstracts, pre-clinical studies, technical reports, and reviews. 2. Materials and Methods The search strategy used in this systematic review was based on the PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) guidelines (http://www.prisma-statement.org) [35]. Clinical questions were formulated according to the PICO framework for evidence based practice [36]. The focused question was: “What is the efﬁcacy of bone augmentation techniques using PDLLA devices compared to traditional materials in terms of qualitative and/or quantitative changes of hard tissue at the augmented sites?” 3 of 14 Table 1. Systematic search strategy. Molecules 2017, 22, 2214 Molecules 2017, 22, 2214 A literature search was carried out on November 2017 by two independent and calibrated reviewers (M.A. and G.C.) using an ad hoc created search string in the database of the National Library of Medicine MEDLINE/PubMed, in the Cochrane Central Register of Controlled Trials, and in the ClinicalTrials.gov website. No language restriction was applied. All interventional studies (either randomized or non-randomized controlled clinical trials) and observational studies (either analytical or descriptive) assessing the efﬁcacy PDLLA devices to bone augmentation techniques were considered. Sinus lift and alveolar preservation procedures were also included. Preclinical studies, technical reports, case reports, review, and conference abstracts were not considered. No restriction on age or number of patients, as well as on the follow-up duration was considered. Studies regarding osteosynthesis techniques and periodontal regeneration were not included. All full text studies were carefully read and analyzed for the eligibility criteria (inclusion/exclusion). Reported results and data were analyzed for qualitative and/or quantitative changes of hard tissue at the augmented sites. The main features of the bibliographic search are reported in Table 1. Systematic Search Strategy Search string: (pdlla OR “poly-D,L-lactic acid” OR “poly(L,D-lactide)” OR “poly-D-L-lactide” OR “poly-D,L-lactide” OR “poly(L/D-lactide)” OR “poly(L,D-lactic acid)” OR “poly-D,L-lactide acid” OR “Atrisorb” OR “Resorb-X” OR “SonicWeld” OR “SonicPins”) AND (oral OR dental OR bone) AND (membrane OR shell OR pin OR barrier OR screw OR mesh) Filter: Humans Language: No restrictions Electronic databases: MEDLINE/PubMed, The Cochrane Central Register of Controlled Trials (CENTRAL), ClinicalTrials.gov Inclusion criteria: - Study design: interventional and observational studies - Population: partially edentulous patients - No restriction on age or number of patients - Healthy individuals (no systemic diseases, no medications affecting platelet and bone functions) - Intervention: bone augmentation using PDLLA devices - Comparison (if appliable): any other augmentation technique or material - Outcome: qualitative and/or quantitative changes of hard tissue at the augmented sites. - Follow up duration: any Exclusion criteria: - Studies regarding osteosynthesis techniques or periodontal regeneration - Conference abstracts, pre-clinical studies, technical reports, and reviews. 3. Results 456 items in MEDLINE/PubMed and 16 in other sources were found after the initial search (14 items in the Cochrane Library, 2 in the ClinicalTrials.gov database). After duplicates were removed, 460 records remained. They were screened on the basis of titles and abstracts for inclusion/exclusion criteria, so that 452 studies were excluded. After full text reading of the remaining eight papers, two of them were excluded because they used a polylactic material different from PDLLA. At the end of the process, six studies published between 2001 and 2016 were included in this systematic review (Figure 1). 4 of 14 4 of Molecules 2017, 22, 2214 Molecules 2017, 22, 22 Figure 1. Flow diagram (PRISMA format) of the screening and selection process. The main characteristics of the included articles are reported in Table 2. Identification Records screened after duplicates and no data items were removed (n = 460) Screening Records excluded (n = 452) Full-text articles assessed for eligibility (n = 8) Eligibility Full-text articles excluded (n = 2) Included Studies included in the present systematic review (n = 6) Records identified through Medline/PubMed database searching (n = 456) Records identified through other database searching (n = 16) Figure 1. Flow diagram (PRISMA format) of the screening and selection process. The main characteristics of the included articles are reported in Table 2 Figure 1. Flow diagram (PRISMA format) of the screening and selection proce Figure 1. Flow diagram (PRISMA format) of the screening and selection process. The main characteristics of the included articles are reported in Table The main characteristics of the included articles are reported in Table 2. 5 of 14 5 of 14 Molecules 2017, 22, 2214 Table 2. Main characteristics of the included articles. RCT: randomized controlled trial; CT: controlled trial; ND: not deﬁnable; CBCT: cone beam computed tomography. Autor, Year Design Patients (Sites) Groups Follow-Up Results Conclusions Gocmen et al., 2016 RCT, Split mouth 10 (20) • Test side: elevation of the maxillary sinus membrane and placement of a PDLLA pin. Control side: elevation of the maxillary sinus membrane and placement of a hyaluronic acid matrix. 6 months Signiﬁcantly higher postoperative mean values of alveolar bone height on both sides compared to the preoperative ones. Signiﬁcantly greater increase of alveolar bone height and reduction in sinus volume on the test side compared to the control side. 100% implant survival rate at both sides. 3. Results There was sufﬁcient bone height to eventually place the implant in both sides of all patients. The two techniques yielded predictable outcomes in implant survival and bone quality. However, the height of alveolar bone and reduction in sinus volume were considerably greater on the PDLLA pin sides. Lie et al., 2015 RCT, Split mouth. 5 (10) • Test side: sinus lift using exclusively a PDLLA membrane ﬁxed with PDLLA pins. Control side: sinus lift using a mixture of inorganic bovine bone substitute material and autologous bone. 6 months Bone formation revealed by CBCT at both sides, but less radiopacity shown at the test sides. Vital new bone at the histological analysis, although less organized and immature, at the test side and a mixture of autogenous and residual bone substitute at the control side. 100% implant survival rate at both sides. The PDLLA membrane proved to be a good and reliable technique to create a stable elevation of the sinus membrane and seemed not interfere with the bone forming process. Burger BW, 2010 CT, Parallel groups ND • Test group: horizontal bone augmentation with PDLLA membrane and pins + particulate bone allograft. Control group: horizontal bone augmentation with collagen membrane + particulate bone allograft. ND About 3 mm increase in bone width measured by computed tomography. No clinical difference in the nature of the grafts and in the bone density between the test and control groups. Unremarkable tissue response. No evidence of prolonged inﬂammation, swelling, or discomfort. There are many advantages to using the resorbable PDLLA pins and foil panels for augmenting alveolar ridge defects: the handling and placing the pins and foil is easy and provides excellent support for the underlining mucoperiosteum for particulate bone grafting. omized controlled trial; CT: controlled trial; ND: not deﬁnable; CBCT: cone beam computed tomography. There are many advantages to using the resorbable PDLLA pins and foil panels for augmenting alveolar ridge defects: the handling and placing the pins and foil is easy and provides excellent support for the underlining mucoperiosteum for particulate bone grafting. The PDLLA membrane proved to be a good and reliable technique to create a stable elevation of the sinus membrane and seemed not interfere with the bone forming process. Conclusions There was sufﬁcient bone height to eventually place the implant in both sides of all patients. The two techniques yielded predictable outcomes in implant survival and bone quality. However, the height of alveolar bone and reduction in sinus volume were considerably greater on the PDLLA pin sides. The PDLLA membrane proved to be a good and reliable technique to create a stable elevation of the sinus membrane and seemed not interfere with the bone forming process. There are many advantages to using the resorbable PDLLA pins and foil panels for augmenting alveolar ridge defects: the handling and placing the pins and foil is easy and provides excellent support for the underlining mucoperiosteum for particulate bone grafting. 6 of 14 Molecules 2017, 22, 2214 Table 2. Cont. Table 2. Cont. Autor, Year Design Patients (Sites) Groups Follow-Up Results Conclusions Santana & de Mattos, 2009 CT, split mouth 22 (56) • Test side: post extractive sites treated with decalciﬁed freeze-dried bone allograft and inorganic bovine bone mineral graft + vascularized periosteal membrane + ﬂuid PDLLA membrane. Control side: post extractive sites treated with decalciﬁed freeze-dried bone allograft and inorganic bovine bone mineral graft + vascularized periosteal membrane. 6 months Ridge dimension change showed a small not statistically signiﬁcant difference in favor of the PDLLA membrane. 7% membrane exposition rate at the test sites. No signs of erythema, edema, swelling, or suppuration in any test or control site. Comparable soft tissue healing at both test or control sites. Fluid PDLLA m suitable for alv in conjunction w grafting. Within present study, i vascularized pe an adequate ch maintaining co coverage and h intention of gra humans. The p optimal levels o absorbable mem however, the us membrane prov clinical beneﬁts Raghoebar et al., 2006 RCT, split mouth 8 (16) • Control side: autologous bone graft ﬁxed with titanium screws. Test side: autologous bone graft ﬁxed with resorbable PDLLA screws. 3–9 months Comparable increase (about 4 mm) of the alveolar ridge width in both test and control sides. PDLLA material still visible at the histological analysis, not in direct contact with the bone and surrounded by ﬁbrous tissue, thin and immature, with a lot of giant cells; no necrosis, hemorrhage, or signiﬁcant inﬂammatory response. 100% implant survival rate at both sides. Resorbable scre ﬁxation devices grafts. Fluid PDLLA membranes can be used in conjunction with a bone graft to treat peri-implant dehiscences and fenestrations. Resorbable screws can be used as ﬁxation devices for autologous bone grafts. However, care must be taken when using these screws because of the presence of the polymer (remnants) after nine months, which could interfere in the sequence of the treatment with endosseous implants. Fluid PDLLA membranes can be used in conjunction with a bone graft to treat peri-implant dehiscences and fenestrations. Conclusions Howeve when using the presence of the nine months, w the sequence of endosseous imp Rosen & Reynolds, 2001 Observational descriptive study (case series) 9 (11) • 11 implants placed in combination with a composite graft of demineralized freeze-dried bone allograft (DFDBA) mixed with freeze-dried bone allograft (FDBA) in a 1:1 ratio, covered with a ﬂuid PDLLA membrane. 4 to 8.5 months Complete coverage in 10/11 sites (success rate 90.9%). No site had inﬂammatory reactions or infection. Vital bone, at the histologic evaluation, in close amalgamation with graft particles, without inﬂammatory inﬁltrate. Fluid PDLLA m conjunction wit peri-implant de fenestrations. Conclusions Conclusions Fluid PDLLA membrane may be suitable for alveolar ridge preservation in conjunction with composite bone grafting. Within the limitations of the present study, it was concluded that the vascularized periosteal membrane was an adequate choice for achieving and maintaining complete soft tissue coverage and healing by primary intention of grafted extraction sockets in humans. The procedures allowed for optimal levels of complete coverage of absorbable membranes during healing; however, the use of a ﬂuid PDLLA membrane provided no additional clinical beneﬁts. Resorbable screws can be used as ﬁxation devices for autologous bone grafts. However, care must be taken when using these screws because of the presence of the polymer (remnants) after nine months, which could interfere in the sequence of the treatment with endosseous implants. p 100% implant survival rate at both sides. Complete coverage in 10/11 sites (success rate 90.9%). No site had inﬂammatory reactions or infection. Vital bone, at the histologic evaluation, in close amalgamation with graft particles, without inﬂammatory inﬁltrate. No site had inﬂammatory reactions or infection. Vital bone, at the histologic evaluation, in close amalgamation with graft particles, without inﬂammatory inﬁltrate. Molecules 2017, 22, 2214 7 of 14 In the study of Rosen & Reynolds (2001) titled “Guided bone regeneration for dehiscence and fenestration defects on implants using an absorbable polymer barrier” [37], PDLLA ﬂuid membranes (Atrisorb) were used to treat peri-implant defects at the time of implant insertion. The purpose of the report was to present consecutive clinical experiences with a poly(D,L-lactide) polymer barrier in combination with a composite bone replacement graft for GBR of dehiscence/fenestration defects during implant placement. Two implant sites requiring ostectomy for exposure of the ﬁxture permitted histologic evaluation of the new bone formation. Nine patients, for a total of 11 peri-implant defects, were treated with a composite graft of demineralized freeze-dried bone allograft (DFDBA) mixed with freeze-dried bone allograft (FDBA) in a 1:1 ratio. The graft was then covered with the PDLLA membrane that overlapped the adjacent osseous structures for 2 to 3 mm. Photographs were taken of the implant and GBR procedure, and clinical measurements were made of the dehiscence/fenestration defects treated. The osseous defects treated included eight dehiscence and three fenestration defects. The success or failure of the GBR procedure was assessed at the second-stage surgery when ﬂaps were reﬂected to place healing abutments. Complete success was deﬁned as coverage of all threads or exposed implant surface. Conclusions Partial success was deemed as incomplete coverage of most threads or exposed implant surfaces with a maximum of two threads or 2 mm of implant surface left uncovered. Failure was deﬁned as no coverage beyond two threads or 2 mm of implant surface. The length of the defects ranged from 2 to 13 exposed threads with an average of 8.5 exposed threads. With the exception of one dehiscence lesion that had partial coverage, all other defects had complete coverage of the implants, representing a 90.9% success rate. The barriers were well tolerated by the tissue with no sites demonstrating any type of tissue reactions or infection. The histologic evaluation of biopsy specimens revealed the presence of viable bone and residual graft particles. Notably absent in all specimens was evidence of an inﬂammatory cell inﬁltrate. The authors emphasize some advantages of this ﬂuid barrier. The polymer system permits repair or modiﬁcation of the barrier if it is inaccurately cut or trimmed, since additional material can be added to the existing barrier. The barrier’s ﬁnal consistency is hard and rigid, adding to its space maintenance capabilities. It takes approximately 5 to 6 months for signiﬁcant degradation of the barrier to begin, and it is probably completely absorbed by 12 months. The authors also highlight some limitations of this material. The ﬁnal consistency of the barrier is hard and ﬁrm, making it susceptible to fracture and premature loss in the presence of untoward occlusal forces. This event occurred in a patient at one implant site, although the clinical outcome was still favorable. In the presence of large defects, premature loss of the barrier might substantially compromise the regenerative outcome. Furthermore, the exposure of the poly(D,L-lactide) barrier may also lead to a premature loss, with regenerative consequences related to the maturational stage of the wound healing process. In the study of Raghoebar et al. (2006) titled “Resorbable Screws for Fixation of Autologous Bone Grafts” [38] PDLLA pins for ﬁxating autologous bone grafts were used. Eight patients were included in this split mouth RCT. In all cases, a two-stage procedure (ﬁrst stage—bone grafting; second stage—placement of implants) was performed bilaterally. Randomly, the grafts were ﬁxed to the alveolar bone with 1.5 mm large titanium screws at one side (Martin Medizin Technik) (control) and with 2.1 mm large PDLLA resorbable screws (Resorb X, Martin Medizin Technik) on the other side (test). Conclusions The bone blocks were ﬁxed with two titanium screws on one side or two resorbable screws on the other side, in a region where no implants were planned to be inserted. The bone width was measured with a caliper. The bone grafts were covered also with a resorbable membrane. After a healing period of at least three months, the implant placement procedure was performed. After reﬂecting the mucoperiosteal ﬂap, the width of the reconstructed alveolar crest was measured and the titanium screws were removed. Using the template for insertion of the implants, in order to avoid areas in which the implants will be inserted, a bone biopsy was taken with a trephine bur including one resorbable screw. In all cases, the bone volume was sufﬁcient. Six months after insertion, the implants were uncovered and another bone biopsy was taken from the area with the other resorbable screw included. The width of the alveolar ridge was between 2 and 4 mm before augmentation and 6 and 8 mm after 8 of 14 Molecules 2017, 22, 2214 reconstruction. There was no difference in the bone width between the two sides. Neither at three nor at nine months were observed differences between both sides. A total of 56 implants were placed in the augmented maxillae. No implants were lost during the follow-up (22.2 ± 4.3 months). Microscopically, three and nine months after reconstruction of the maxilla with autologous bone (most of) the PDLLA material was still visible. The material was shown to be biocompatible as no inﬂammatory response was observed both at the three- and nine-month evaluation periods. It must be emphasized that PDLLA screws did not interfere in the incorporation of the grafts and on bone viability and/or quality. The PDLLA screws, whose contours were clearly visible in all biopsies, were encapsulated by a thin and immature ﬁbrous tissue capsule containing many giant cells in direct contact with the PDLLA material as well as inﬁltrating in areas with fragmented PDLLA. PDLLA particles were observed within the giant cells. The adaptation of the biodegradable screws for ﬁxation of the bone grafts resulted as uncomplicated as with titanium screws. Conclusions The authors emphasize the advantages of using a biodegradable material for osteosynthesis over metals: no need of material removal; no risk that the head of the screw can protrude as the bone grafts heal and bone resorption occurs, causing discomfort, soft-tissue dehiscence, infection, and even graft failure [39]; no risk of compromission of the implant area, possible when the metallic screws, covered by bone, were positioned in the region where the implants had to be inserted. Furthermore, although the small sample size, the authors highlight that no signiﬁcant difference, and even no tendency, in remodeling of the grafts ﬁxed with either titanium or resorbable screws could be observed. However, they also underline a signiﬁcant ﬁnding concerning PDLLA resorbability. After nine months, in fact, screw remnants were still present, surrounded by foreign body cells. Although no signiﬁcant inﬂammatory signs were visible and no patient presented any kind of clinical or radiographic manifestation suggestive of a more severe inﬂammatory reaction in response to PDLLA than titanium screws, this ﬁnding suggests caution using this resorbable material as ﬁxation devices when endosseous implant insertion is planned. The presence of the connective tissue in bone–screw interface does not represent an obstacle to the placement of implants. However, whether the maintenance of the polymer in this speciﬁc experimental period can interfere during the preparation of the implant site and the effects of this manipulation cannot be predicted. Considering this observation, the authors afﬁrm that the resorbable screws would be of great use, especially in large reconstructions, if they can be positioned in areas where they certainly will not interfere with the planning of the endosseous implants. p g p In the study of Santana & de Mattos (2009) titled “Efﬁcacy of Vascularized Periosteal Membranes in Providing Soft Tissue Closure at Grafted Human Maxillary Extraction Sites” [40], a ﬂuid PDLLA membrane (Atrisorb) a total of 56 post-extractive sites in 22 patients with a split mouth design was used. The objective of this study was to evaluate the efﬁcacy of a ﬂap design, based on the extension of palatal tissues, to obtain and maintain soft tissue coverage over grafted extraction sockets with or without the use of an absorbable ﬂuid PDLLA membrane. In the control side, post-extractive sites were treated with a 1:1 mixture of decalciﬁed freeze-dried bone allograft material (DFDBA) and an inorganic bovine bone mineral (Bio-Oss™), covered with the vascularized periosteal membrane. Conclusions The test side received the same graft material and the vascularized periosteal membrane, but the sites were also covered with a ﬂuid PDLLA membrane. The measurements of the crestal bone width were made from the crest of the buccal bony wall to the crest of the palatal bony wall, with a calibrated periodontal probe at a point corresponding to half the mesiodistal socket diameter. The measurements were made at two times: at the time of the surgery, just prior to the graft placement, and six months after tooth extraction, after the elevation of full-thickness buccal and palatal ﬂaps to expose the alveolar crest. Baseline measurements of the ridge dimensions (BCW) revealed a mean of 7.9 ± 0.9 mm for the control sites and 8.2 ± 0.8 mm for the experimental sites. Repeated measurements obtained six months after tooth extraction showed that the test and control groups performed equally well. Mean BCW in the controls was 6.9 ± 0.7 mm, and in the experimental sites it was 7.5 ± 0.8 mm. This small difference did not reach statistical signiﬁcance. The results of the present study showed that both techniques were effective in obtaining and maintaining soft tissue closure over grafted extraction sockets. The presence 9 of 14 Molecules 2017, 22, 2214 of a barrier membrane did not increase the incidence of soft tissue dehiscence, and elevated levels of soft tissue closure were maintained throughout the study period. Membrane exposure was observed in 7% of the test sites. In view of the reported variations in frequencies of barrier exposure following GBR procedures reported in the literature, as well as ﬁndings of reduced bone regeneration in cases of early membrane exposure [13,41] the results of the present study suggest that the vascularized periosteal membrane technique holds promise in optimizing the clinical results of GBR protocols. No signs of erythema, edema, swelling, or suppuration were noticed in the soft tissues of the experimental group. Soft tissue healing at sites treated with the absorbable membrane did not appear to be different from that observed at the control sites. Moreover, in the few areas where the membranes were exposed during healing, the exposed parts of the membrane margins disappeared concurrently with healing of the soft tissues. No signs of infection or suppuration were observed in exposed membrane sites, suggesting that the material was well tolerated by the tissues. Conclusions The authors conclude that this material, in addition to GTR, may also be suitable for alveolar ridge preservation in conjunction with composite bone grafting and that, within the limitations of the study, the vascularized periosteal membrane was an adequate choice for achieving and maintaining complete soft tissue coverage and healing by primary intention of grafted extraction sockets in humans. The procedures allowed for optimal levels of complete coverage of absorbable membranes during healing; however, the use of an absorbable membrane provided no additional clinical beneﬁts. In the study of Burger B. W. (2010) titled “Use of Ultrasound-Activated Resorbable Poly-D-L-lactide Pins (SonicPins) and Foil Panels (Resorb-X) for Horizontal Bone Augmentation of the Maxillary and Mandibular Alveolar Ridges” [42] have been used. The aim of this study was to determine the efﬁcacy of 50% poly-D-lactide and poly-L-lactide pins (SonicPins, KLS Martin) and foil panels (Resorb-X, KLS Martin) in conjunction with a particulate bone allograft (Puros, RTI; Biologics; Alachura, FL, USA) in patients in need of at least 3 mm of horizonal bone augmentation in multiple sites of maxilla and mandible. Although the authors also report in the text the use of both a PDLLA membrane and a collagen membrane (OsseoGuard GTM, Biomed, 3i West Palm, FL, USA), in conjunction with the bone graft for the resolution of horizontal ridge defects, the paper is not clearly structured with a controlled study design (experimental groups are not punctually described and comparative outcome measures are not reported at all). The authors extensively describe the surgical procedure, together with the advantages of the PDLLA pins and membranes. The sites for pin placement were prepared with a drill that penetrated at least through the outer bony cortex; the pin is pushed into a funnel-shaped pilot hole with sonic frequency vibration: the edges of the pin rub on the bone, causing friction to bring the polymer to a liquid state. The pilot hole is smaller than the pin, causing the polymer to ﬂow into the trabecular spaces of the bone. When the sonic frequency is stopped, these liqueﬁed portions become hard again in seconds. When welding the resorbable SonicPins to the bone, the pins were extremely easy to handle and it took very little time to place them, even in the posterior maxilla, where access is limited. Conclusions The sonic pins were very secure, did not fracture or break, and did not have to be replaced with a larger diameter pin. It was also easy to shorten the pins by resonication. The Resorb-X foil panels did not need any additional support: although it was extremely thin, the foil provided its own rigidity and support after cooling. Once the foil is heated, it can be adapted to the bone surface and shaped to the desired position. Once cooled, the material turns rigid again and reliably retains its shape. The foils used in this study were 0.1 mm thick, which provided excellent support for the particulate bone graft and to tent the mucoperiosteum. Tissue response to the PDLLA devices was unremarkable: there was no evidence of prolonged inﬂammation, swelling, or discomfort at any of the operative sites. When comparing preoperative with postoperative computer tomographic scans, there was an approximately 3 mm increase in bone width. Upon entering the grafted sites, there was sufﬁcient bone width to place the implants. In conclusion, the authors underline some advantages with the use of these resorbable PDLLA foil panels and pins: the easy handling and placing of pins and foil, the excellent support provided by pins and foils to the mucoperiosteum, above the underlying particulate bone grafting; the possibility of shaping foils to the desired dimension for adequate augmentation. Molecules 2017, 22, 2214 10 of 14 We choose to consider, in a more extensive way, also the maxillary sinus elevation among the bone augmentation procedures. For this reason, we included in the present review two recent studies on the use of such devices for the maxillary sinus elevation. We choose to consider, in a more extensive way, also the maxillary sinus elevation among the bone augmentation procedures. For this reason, we included in the present review two recent studies on the use of such devices for the maxillary sinus elevation. In their study titled “Elevation of the Maxillary Sinus Membrane for De Novo Bone Formation: First Results of a Prospective Study in Humans” [43], Lie et al. (2015) used PDLLA membranes to perform an elevation of the maxillary sinus membrane. The aim of this study was the clinical and histological investigation of bone regeneration and integration of a well characterized inorganic bovine bone substitute material (Bio-Oss™; Geistlich Biomaterials GmbH, Wolhusen, Switzerland) mixed with autogenous bone in comparison to elevation of the sinus membrane alone. Conclusions Five patients were included in this split mouth RCT. One maxillary side was augmented using the standard augmentation procedure with a mixture of autogenous and xenogenous bone. On the contralateral side the Schneiderian membrane of the maxillary sinus was lifted and stabilized with a degradable perforated PDLLA membrane (Resorb-X, KLS Martin) and PDLLA pins (SonicPins, KLS Martin). Six months after the sinus ﬂoor augmentation, six implants in each patient were positioned, with a total of 30 implants. Pre- and post-operative cone-beam computed tomography (CBCT) scans where compared and the gain in bone volume was measured using a digital program. During the implant insertion, a trephine bur was used for the preparation, so it was possible also to take bone probes for histological evaluation. After six months, sufﬁcient bone was present in both sides at the radiological evaluation. However, the non-augmented side showed less opacity, which suggested less bone density or less presence of mature bone. However, primary stability of the implants was achieved in all cases independently of the augmentation technique applied. All bone specimens showed new bone formation. However, the bone of the experimental side was less organized and immature. The bone biopsies of the conventional side showed as expected the combination of xenogenous bone, embedded in bone marrow and newly formed bone. The biopsies from the experimental side showed new bone formation and osteoblast and osteoclast activity as signs of an active bone forming and remodelling process. Implant success rates were 100% on both sides one year after the prosthodontic rehabilitation. All implants could be loaded by prostheses in a conventional way (bar-retained overdentures) and are successful after the follow up time. All patients reported comfort with the dentures and were fully satisﬁed. Based on the randomization, patients did not mention differences between both sides of the maxilla. Prosthetic follow up differed from 12 months post-loading in the ﬁrst study patient to 6 months post-loading in the last patient. The authors concluded that the use of a resorbable PDLLA membrane proved to be a good and reliable technique to create a stable elevation of the sinus membrane. The major advantage of the technique presented is the absence of any kind of donor site morbidity (autogenous bone) and there is no risk of infectious proteins (xenogenous bone substitutes). Conclusions PDLLA membrane did not interfere with the bone forming process, which resembles a callus-based bone formation in a space surrounded by bony walls. The perforated mesh design of the membranes used seems to be logical to allow vascular ingrowth and the material degrades without generation of any crystalline remnants [44]. In the study of Gocmen et al. (2016) titled “Hyaluronic Acid versus Ultrasonic Resorbable Pin Fixation for Space Maintenance in Non-Grafted Sinus Lifting” [45], PDLLA pins were used to perform an elevation of the sinus membrane. The aim of this study was to compare hyaluronic acid with ultrasonic resorbable pins for their ability to maintain intrasinusal space. 10 patients were included in this split mouth RCT. One maxillary side was augmented with the elevation of the sinus membrane and the positioning of a hyaluronic acid matrix (Hyaloss Matrix, Anika Therapeutics, Bedford, MA, USA). On the contralateral side, the Schneiderian membrane of the maxillary sinus was lifted and stabilized with a degradable PDLLA pin (Sonic-Pin Rx, KLS Martin, Muhlheim, Germany) of 11 to 17 mm in length, ﬁxed 2 mm above the bone window used to perform the sinus lift. CBCTs were taken before the surgery and six months after. The mean postoperative height of alveolar bone values were 9.6 ± 3.4 mm on the Sonic-Pin Rx side and 6.4 ± 2.6 mm on the hyaluronic acid side. The postoperative mean height of alveolar bone values were signiﬁcantly greater than the preoperative mean values 11 of 14 Molecules 2017, 22, 2214 on the two sides. The mean increases in height of alveolar bone and reduction of sinus volume on the Sonic-Pin side were signiﬁcantly greater than those on the hyaluronic acid side. Patients were treated with two implants in each side, for a total of 40 implants placed. The bone quality at the moment of the implant insertion was of type II or higher in all the sites. A late-loading protocol was followed, and the mean healing time for abutment loading was 16.4 ± 2.5 weeks. At six months, all implants were clinically stable, and the deﬁnitive prostheses were functional, with a survival rate of 100%. In conclusion, there was sufﬁcient bone height to eventually place the implant in the right and left sides of all patients. The two techniques yielded predictable outcomes in implant survival and bone quality. Conclusions However, the height of alveolar bone and reduction in sinus volume were considerably greater on the PDLLA pin sides. This might be due to the features of the resorbable pins as a rigid barrier or the weak mechanical properties of bone formation on the hyaluronic acid side. 4. Discussion and Conclusions PDLLA devices have been proposed for bone augmentation procedures in different forms, based on the advantageous possibility of enhancing the biomechanical, rheological, thermal, and biological properties of this material by modulating the ratio of D and L isomers in its formulations. Despite a larger presence of pre-clinical studies as well as of clinical application of PDLLA devices for other applications, such as osteosynthesis techniques, clinical data on bone augmentation procedures are very scarce. Only six clinical studies about PDLLA devices for bone augmentation were found. One of them was a descriptive study (case series), ﬁve were controlled studies (three randomized). PDLLA was used in the form of PDLLA foils, screws, and pins. An evident heterogeneity in clinical indications and augmentation techniques used with the aid of the PDLLA devices was found: alveolar preservation of post-extraction sites by PDLLA foils, bone block ﬁxation by PDLLA screws, sinus lift using PDLLA foils or pins, and correction of peri-implant dehiscences and fenestrations by PDLLA membranes and pins. Such a variability impeded any qualitative or quantitative comparison among the studies. About the clinical use of PDLLA devices the authors emphasize the ease of handling, absence of a re-entry phase, moldability of foils, good soft-tissue response. The authors point out also some drawbacks including the risk of membrane exposition, a prolonged adsorbability, and, from a histological point of view, a tendency to the ﬁbrous encapsulation of the PDLLA devices (although the clinical signiﬁcance of this ﬁnding is unclear). In conclusion, the results from the available studies are inconclusive about the clinical efﬁcacy of PDLLA devices for bone augmentation procedures, mainly because of the high heterogeneity and the lack of sufﬁciently powered and appropriately designed trials. Well-designed randomized controlled trials of sufﬁcient size comparing the use of PDLLA foils and pins with conventional membranes for bone augmentation procedures are strongly encouraged in order to understand the real clinical beneﬁts/drawbacks of this technique. Acknowledgments: Institutional funds from the University of Campania “Luigi Vanvitelli” were used for covering the article processing charge in open access. Author Contributions: M.A. conceived and designed the study; M.A. and G.C. performed the systematic research; L.N. analyzed and elaborated the data; M.A., L.N., G.C. and L.G. wrote the paper. L.G. coordinated the study e revised the paper. Conﬂicts of Interest: The authors declare no conﬂict of interest. 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Santana, R.B.; de Mattos, C.M. Efﬁcacy of vascularized periosteal membranes in providing soft tissue closure at grafted human maxillary extraction sites. Int. J. Oral Maxillofac. Implants 2009, 24, 81–87. [PubMed] 41. Lekholm, U.; Becker, W.; Dahlin, C.; Becker, B.; Donath, K.; Morrison, E. The role of early versus late removal of GTAM membranes on bone formation at oral implants placed into immediate extraction sockets. An experimental study in dogs. Clin. Oral Implants Res. 1993, 4, 121–129. [CrossRef] [PubMed] 42. Burger, B.W. Use of ultrasound-activated resorbable poly-D-L-lactide pins (SonicPins) and foil panels (Resorb-X) for horizontal bone augmentation of the maxillary and mandibular alveolar ridges. J. Oral Maxillofac. Surg. 2010, 68, 1656–1661. [CrossRef] [PubMed] 43. Lie, N.; Merten, H.A.; Meyns, J.; Lethaus, B.; Wiltfang, J.; Kessler, P. Elevation of the maxillary sinus membrane for de-novo bone formation: First results of a prospective study in humans. J. Cranio-Maxillofac. Surg. 2015, 43, 1670–1677. [CrossRef] [PubMed] 43. Lie, N.; Merten, H.A.; Meyns, J.; Lethaus, B.; Wiltfang, J.; Kessler, P. Elevation of the maxillary sinus membrane for de-novo bone formation: First results of a prospective study in humans. J. Cranio-Maxillofac. Surg. 2015, 43, 1670–1677. [CrossRef] [PubMed] 43. Lie, N.; Merten, H.A.; Meyns, J.; Lethaus, B.; Wiltfang, J.; Kessler, P. Elevation of the maxillary sinus membrane for de-novo bone formation: First results of a prospective study in humans. J. Cranio-Maxillofac. Surg. 2015, 43, 1670–1677. [CrossRef] [PubMed] 14 of 14 14 of 14 Molecules 2017, 22, 2214 44. Heidemann, W.; Jeschkeit, S.; Rufﬁeux, K.; Fischer, J.H.; Wagner, M.; Kruger, G.; Wintermantel, E.; Gerlach, K.L. References Degradation of poly(D,L)lactide implants with or without addition of calciumphosphates in vivo. Biomaterials 2001, 22, 2371–2381. [CrossRef] 44. Heidemann, W.; Jeschkeit, S.; Rufﬁeux, K.; Fischer, J.H.; Wagner, M.; Kruger, G.; Wintermantel, E.; Gerlach, K.L. Degradation of poly(D,L)lactide implants with or without addition of calciumphosphates in vivo. Biomaterials 2001, 22, 2371–2381. [CrossRef] 45. Gocmen, G.; Atah, O.; Aktop, S.; Sipahi, A.; Gonul, O. Hyaluronic acid versus ultrasonic resorbable pin ﬁxation for space maintenance in non-grafted sinus lifting. J. Oral Maxillofac. Surg. 2016, 74, 497–504. [CrossRef] [PubMed] © 2017 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) li (htt // ti /li /b /4 0/) 45. Gocmen, G.; Atah, O.; Aktop, S.; Sipahi, A.; Gonul, O. Hyaluronic acid versus ultrasonic resorbable pin ﬁxation for space maintenance in non-grafted sinus lifting. J. Oral Maxillofac. Surg. 2016, 74, 497–504. [CrossRef] [PubMed] © 2017 by the authors. Licensee MDPI, Basel, Switzerland. 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https://openalex.org/W3211296373	OpenAlex	Open Science	CC-By	2,021	Comunidades Criativas e o designer como mediador social	Larissa Vargas Teixeira	Portuguese	Spoken	2,782	5,176	design, comunidades criativas, inovação social, mediação design, comunidades criativas, inovação social, mediação Em vista das imensuráveis mudanças que vêm ocorrendo no cenário mundial atual, o propósito deste artigo é compreender os aspectos em que o design pode promover transformações significativas dentro do contexto das comunidades criativas. Estas transformações estão associadas ao contínuo progresso do design em relação às inovações sociais que, por sua vez, estão pautadas diretamente por uma mudança de comportamento, caracterizada pelo desenvolvimento de novas estratégias, conceitos e organizações que atendem a necessidades sociais de todos tipos, no intuito de fortalecer a sociedade na qual está inserida. Para isto, as comunidades criativas se qualificam como os meios onde essas inovações sociais são aplicadas, pois são formadas por indivíduos que se organizam com a finalidade de construir novas soluções para o seus contextos. Nesse cenário, o designer torna-se um importante facilitador, podendo auxiliar em campos de conhecimentos distintos e, consequentemente, promovendo novas possibilidades e novos caminhos. design, creative communities, social innovation, mediation In view of the immeasurable changes that have been taking place in the current world scenario, the purpose of this article is to understand the aspects in which design can promote significant transformations within the context of creative communities. These transformations are associated with the continuous progress of design in relation to social innovations, which, in turn, are guided directly by a change in behavior, characterized by the development of new strategies, concepts and organizations that meet social needs of all types, in order to strengthen the society in which it is inserted. For this, creative communities qualify as the means where these social innovations are applied, this is because they are formed by individuals who organize themselves with the purpose of building new solutions for their context. In this scenario, the designer becomes an important facilitator, being able to assist in different fields of knowledge and, consequently, promoting new possibilities and new paths. Comunidades Criativas e o designer como mediador social Creative Communities and the designer as a social mediator Larissa Vargas Teixeira, Carolina Iuva de Mello 1 Introdução As comunidades criativas são caracterizadas como grupos de indivíduos que ao se depararem com obstáculos buscam contribuir conjuntamente na construção de novas soluções a partir do seu contexto e os recursos disponíveis. Para estimular a formação dessas comunidades, o designer pode atuar nesse âmbito para desenvolver e efetivar soluções habilitantes que contribuam para a cooperação entre os membros da comunidade. No entanto, ainda há uma imensa barreira a ser ultrapassada, isso porque, o design ainda é popularmente retratado como uma profissão isolada das discussões sociais. Anais do 10º CIDI e 10º CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brasil \| 2021 Proceedings of the 10th CIDI and 10th CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brazil \| 2021 Anais do 10º CIDI e 10º CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brasil \| 2021 Anais do 10º CIDI e 10º CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brasil \| 2021 Proceedings of the 10th CIDI and 10th CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brazil \| 2021 Proceedings of the 10th CIDI and 10th CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brazil \| 2021 Proceedings of the 10th CIDI and 10th CONGIC Kelli C.A.S. Smythe, Rafael de Castro Andrade (orgs.) Sociedade Brasileira de Design da Informação – SBDI Curitiba \| Brazil \| 2021 1826 L. V. Teixeira & C. I. Mello \| Comunidades Criativas e o designer como mediador social Muitos dos projetos que visam o desenvolvimento de comunidades ficam aquém de seus objetivos iniciais devido à falta de participação real das pessoas envolvidas. Segundo Geilfus (2002), isso ocorre porque muitos técnicos ainda veem o desenvolvimento como um processo linear simples, onde se passa da situação “A” para a situação “B” em linha reta, no qual a participação se torna uma “perda de tempo” (Geilfus, 2002). 2 Comunidades criativas e soluções habilitantes A essência das comunidades criativas está na interação colaborativa entre diferentes atores no intuito de solucionar problemas em seu próprio território. Estas comunidades funcionam como impulsionadoras de inovações sociais, uma vez que empregam sua criatividade de forma colaborativa, utilizando elementos do seu cotidiano para resolver suas adversidades, implicando, muitas vezes, na criação de soluções sustentáveis e criativas. Nesse sentido, Federizzi (2014) identifica três elementos que compõem o conceito de comunidades criativas, sendo eles: visão social, pensamento em rede e serviços colaborativos. “Esses elementos podem ser encarados como estágios de maturação de um serviço, mas que se movimentam de forma sistêmica, isto é, quanto mais um serviço colaborativo for utilizado, mais fortalecerá a solução proposta, permitindo ajustes e correções no caminho“ (Federizzi, 2014, p. 3). A essência das comunidades criativas está na interação colaborativa entre diferentes atores no intuito de solucionar problemas em seu próprio território. Estas comunidades funcionam como impulsionadoras de inovações sociais, uma vez que empregam sua criatividade de forma colaborativa, utilizando elementos do seu cotidiano para resolver suas adversidades, implicando, muitas vezes, na criação de soluções sustentáveis e criativas. Nesse sentido, Federizzi (2014) identifica três elementos que compõem o conceito de comunidades criativas, sendo eles: visão social, pensamento em rede e serviços colaborativos. “Esses elementos podem ser encarados como estágios de maturação de um serviço, mas que se movimentam de forma sistêmica, isto é, quanto mais um serviço colaborativo for utilizado, mais fortalecerá a solução proposta, permitindo ajustes e correções no caminho“ (Federizzi, 2014, p. 3). [...] as comunidades criativas resultam de uma original combinação de demandas e oportunidades. As demandas, como vimos, são sempre criadas por problemas da vida cotidiana contemporânea. As oportunidades se manifestam a partir de diferentes combinações de três elementos básicos: a existência (ou ao menos a memória) das tradições; a possibilidade de utilizar (de uma forma apropriada) uma série de produtos, serviços e infra-estruturas; a existência de condições sociais e políticas favoráveis (ou pelo menos capazes de aceitar) o desenvolvimento de uma criatividade difusa (Manzini, 2008, p. 65). Diante disso, Manzini (2008) destaca o que chama de soluções habilitantes, que se referem à atividade de criar serviços para encorajar a participação de membros de comunidades de modo colaborativo. As soluções habilitantes são definidas como sistemas de produtos, serviços, meios de comunicação ou um conjunto de todos eles para tornar comunidades criativas mais acessíveis, eficazes, atraentes e replicáveis. 1 Introdução Portanto, faz-se necessário estabelecer premissas que difundam o papel fundamental do designer nas ações em prol do desenvolvimento, introduzindo as maneiras adequadas para que ele possa se inserir neste meio como um mediador social. Assim, o presente artigo busca apresentar uma revisão bibliográfica acerca dos temas comunidades criativas e inovação social, de modo a compreender melhor as maneiras pelas quais o designer pode atuar como mediador em projetos sociais de cunho participativos, utilizando como ferramenta projetual o design de informação. 2 Comunidades criativas e soluções habilitantes O processo corrente estabelecido pelas soluções habilitantes conta com quatro níveis. São estes: (1) Reconhecimento como comunidade; (2) Desenvolvimento de habilidades sociais, criativas, colaborativas, conhecimento distribuído, etc; (3) Autonomia da comunidade, criando novas oportunidades e soluções de problemas; ) Tornar a comunidade criativa mais eficaz, atraente, eficiente e replicável. (4) Tornar a comunidade criativa mais eficaz, atraente, eficiente e replicável. Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021 Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021 1827 L. V. Teixeira & C. I. Mello \| Comunidades Criativas e o designer como mediador social Desse modo, conforme indicam Freire e Oliveira (2017), para que as soluções habilitantes funcionem em comunidades criativas, deve-se partir, no nível um (1), de uma comunidade que se reconheça como tal e de pessoas que já se associam por determinados interesses. Assim, o segundo nível (2) consiste em desenvolver qualidades necessárias para uma comunidade criativa: sociabilidade, colaboração, criatividade, comunicação e conhecimento distribuído. Posteriormente, para o nível três (3), essa comunidade deve alcançar autonomia, de forma que consiga funcionar sem a presença do designer e, principalmente, criando novas oportunidades e solucionando problemas. E então, o passo final (4) seria utilizar estas soluções habilitantes para tornar a comunidade criativa mais eficiente, eficaz, atraente e replicável, conforme previsto na teoria. 3 O designer como mediador em contextos de inovação social Mello \| Comunidades Criativas e o designer como mediador social As soluções incorporadas por Cipolla (2017) refletem o conceito de uma criatividade difusa, oferecendo para as comunidades requeridas uma abordagem focada no processo de obtenção de conhecimento em design, propondo um papel de aprendizagem social e distribuída. Nesse sentido, o profissional pode ver a sua relevância contestada, no entanto, não se trata de questionar a função do designer, mas sim de estabelecer um novo tipo de organização, atribuída em rede, para torná-lo apto a interpretar e fomentar estes processos. Geralmente, os casos onde o designer passa a desenvolver esta mediação nascem a partir de demandas, problemas da vida cotidiana contemporânea ou, ainda, oportunidades. Esses casos são fruto de um grupo de pessoas que foram capazes de dar vida a essas soluções inovadoras, e fizeram isso recombinando o que já existe, sem esperar mudança geral no sistema (Manzini, 2008). Por isso, essas pessoas, capazes de reorganizar elementos já existentes em novas e significativas combinações, ao inventarem, aprimorarem e gerenciarem soluções inovadoras para novos estilos de vida são reconhecidas como comunidades criativas (Meroni, 2007). Nesse sentido, o método colaborativo desenvolvido pelo designer em comunidades criativas também pode ser entendido como um recurso de design informativo, visto que, a maneira como os dados e informações são apresentados é o que define a boa interpretação de um projeto, tornando-o atrativo e reduzindo os erros no processamento (Frascara, 2011). Frascara (2011) também ressalta que o objetivo do design de informação é assegurar a efetividade da comunicação a partir da facilitação de processos de percepção, compreensão e uso da informação apresentada. Nesse cenário, o profissional de design incorpora a importante função de facilitar a recepção e interpretação das inovações que chegam até a comunidade, permitindo uma ação mais eficiente. Assim, o designer deve ser capaz de contextualizar e globalizar, desenvolvendo soluções que relacionem favoravelmente todos os atores e/ou grupos sociais envolvidos, direta ou indiretamente no projeto. Conforme ressaltado por Meroni (2007), toma-se o caminho onde o designer se torna um colaborador para a inovação social, utilizando suas competências profissionais para ajudar as comunidades na tomada de decisões estratégicas, a fim de projetar em conjunto seu futuro, assumindo, assim, um importante papel na sociedade contemporânea. 3 O designer como mediador em contextos de inovação social A inovação social é o resultado do conhecimento aplicado a necessidades sociais por meio da participação e da cooperação de todos os atores envolvidos, gerando novas e duradouras soluções para grupos sociais, comunidades ou para a sociedade em geral. Segundo Manzini (2008, p. 61), a inovação social [...] refere-se às mudanças no modo como indivíduos ou comunidades agem para resolver seus problemas ou criar novas oportunidades. Tais inovações são guiadas mais por mudanças de comportamento do que por mudanças tecnológicas ou de mercado, geralmente emergindo através de processos organizacionais ‘de baixo para cima’ em vez daqueles ‘de cima para baixo’. Design implica mais que o mero exercício de habilidades técnicas. Em contextos de inovação social, os designers podem atuar como experts, que, graças a habilidade com ferramentas e métodos projetuais, podem ajudar outros profissionais a orientar suas iniciativas; ou podem ser parte da solução, justamente por serem os atores sociais que, mais do que qualquer outro, lidam com as interações cotidianas dos seres humanos com seus artefatos (Manzini, 2008). Nesse aspecto, Nussbaumer e Ross (2011, p. 50) afirmam que: O ato de mediar, se assumido como prática social, possibilita recuperar a capacidade de ‘agência’ do ator, enquanto produtor do mundo, sem desconsiderar os condicionantes que lhe impõem o contexto sócio - histórico e institucional no qual opera. Nesta perspectiva, o mediador não é somente um intermediário, mas também intervém ativamente na produção de significados dos mundos que interconecta, dos bens que mobiliza e dos papéis que assume; o mesmo cabe para os mediados. Assim, o designer que atua como agente de mediação social se torna capaz de potencializar soluções às comunidades que acompanha. Isso ocorre porque ele não só atua no nível do projeto, mas no processo de constituição e evolução de um determinado contexto por meio de ações coletivas (Del Gaudio, 2017). A importância de uma atuação em rede para o desenvolvimento e promoção da inovação social pelo design é a necessidade de refletir sobre o papel da prática profissional do designer, bem como seu quadro teórico, diante do reconhecimento do valor de soluções auto-organizadas pelos cidadãos que questionam os modos hegemônicos de produção e consumo (Cipolla, 2017, p. 83). Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021 1828 L. V. Teixeira & C. I. Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021 4 Considerações finais O designer pode atuar como um potencializador de mudanças sociais, especialmente em contextos de inovações sociais propiciadas por comunidades criativas. Contudo, para propor soluções para uma comunidade, é necessário estar verdadeiramente envolvido com ela, conhecer seus problemas, sua rotina, os processos que envolvem o seu trabalho e os valores do seu território. Desse modo, o design de informação é imprescindível para assegurar um sistema de comunicação eficiente, acessível e completo. Para isto, a participação dos designers em projetos de cunho social prevê uma postura de colaboração e não de comando, em uma 1829 L. V. Teixeira & C. I. Mello \| Comunidades Criativas e o designer como mediador social abordagem de ‘design ao lado’, trabalhando como um mediador entre as necessidades das comunidades criativas e os demais atores da sociedade. Sustenta-se, então, a figura do designer como operador para gerar relações que levem os diferentes atores a participar, pois possui habilidades inestimáveis para o desenvolvimento de relações entre os diferentes atores envolvidos. Por fim, reafirma-se a relevância desta revisão bibliográfica, uma vez que visa estabelecer as alegações que auxiliarão na criação de cenários promotores de qualidade das relações interpessoais, sendo que o sucesso dessas relações é apoiado na maneira como elas são nutridas ao longo do processo. Referências Cipolla, C. (2017). Ecovisões sobre Design para a Inovação Social. In: Alfredo Jefferson de Oliveira; Carlo Franzato; Chiara Del Gaudio. (Org.). Ecovisões Projetuais. Pesquisas em Design e Sustentabilidade no Brasil. 1ª ed. São Paulo: Blucher, 1, 83-86. Del Gaudio, C.(2017) Os desafios para o design no âmbito social e as perspectivas futuras: o conceito de infraestruturação e a redefinição do papel do designer. In: Alfredo Jefferson de Oliveira, Carlo Franzato, Chiara Del Gaudio. (Org.). Ecovisões projetuais: pesquisas em design e sustentabilidade no Brasil. 1ª ed. São Paulo: Blucher, 1, 65-80 Federizzi, C. L. (2014). Comunidades criativas para impulsionar a participação colaborativa na cidade. Blucher Design Proceedings, 11° P&D Design, 1(4), 1-12. Frascara. J. (2011). ¿Qué es el diseño de información? Buenos Aires: Ediciones Infinito. Freire, K.; Oliveira, C. M. M. (2017). Soluções habilitantes para formação de comunidades criativas: um caminho possível do design para inovação social. In: Amilton J. V. Arruda. (Org.). Design e Inovação Social. 1ed.São Paulo: Blucher, 1, 109-131. Geilfus, F. (2002). 80 Herramientas para el desarrollo participativo: diagnóstico, planificación, monitoreo, evaluación. San José: IICA Sede Central. Manzini, E. (2008). Design para a Inovação Social e Sustentabilidade: comunidades criativas, organizações colaborativas e novas redes projetuais. Rio de Janeiro: E-papers. Meroni, A. (2007). Creative Communities. People lnventing Sustainable Ways of Living. Milan: Edizioni Polidesign. Nussbaumer, B.; Ross, C. C. (2011). Mediadores sociales: en la producción de prácticas y sentidos de la política pública. Buenos Aires: Fundación CICCUS. Agradecimento Fundo de Incentivo de Extensão da Universidade Federal de Santa Maria (FIEX/UFSM). Sobre as autoras Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021 Larissa Vargas Teixeira, Graduanda, UFSM, Brasil <larissavargast@hotmail.com> Carolina Iuva de Mello, Dra., UFSM, Brasil <carolinaiuva@gmail.com> Larissa Vargas Teixeira, Graduanda, UFSM, Brasil <larissavargast@hotmail.com> Carolina Iuva de Mello, Dra., UFSM, Brasil <carolinaiuva@gmail.com> Anais do 10º Congresso Internacional de Design da Informação \| CIDI 2021 Proceedings of the 10th Information Design International Conference \| CIDI 2021	4,928
https://openalex.org/W3203940082	OpenAlex	Open Science	CC-By-SA	2,021	PEMBERDAYAAN MASYARAKAT DALAM PEMBUATAN BEDENG PERSEMAIAN PADA KOMPEPAR PASIR BATANG TAMAN NASIONAL GUNUNG CIREMAI, KABUPATEN KUNINGAN	Toto Supartono	Indonesian	Spoken	2,574	6,795	ABSTRAK Persemaian merupakan salah satu sarana penting dalam kegiatan pemulihan ekosistem karena dapat berperan sebagai sumber perbanyakan bibit. Tujuan pengabdian masyarakat ini adalah membangun persemaian di Zona Pemanfaatan Taman Nasional Gunung Ciremai yang di dalamnya terdapat kelompok masyarakat yang mengelola obyek wisata. Kompepar Pasir Batang telah dijadikan mitra dalam kegiatan pengabdian masyarakat. Metode yang digunakan adalah penyampaian materi dan implementasi lapangan. Persemaian dapat terbangun sesuai dengan yang diharapkan. Hasil kegiatan menunjukkan bahwa masyarakat memberikan respon positif terhadap kegiatan pengabdian kepada masyarakat yang dilakukan oleh lembaga pendidikan. Kata Kunci: gunung ciremai, pengabdian, persemaian, taman nasional. PEMBERDAYAAN MASYARAKAT DALAM PEMBUATAN BEDENG PERSEMAIAN PADA KOMPEPAR PASIR BATANG TAMAN NASIONAL GUNUNG CIREMAI, KABUPATEN KUNINGAN (COMMUNITY EMPOWERMENT IN THE BUILDING OF NURSERY BED AT KOMPEPAR OF PASIR BATANG IN GUNUNG CIREMAI NATIONAL PARK, KUNINGAN REGENCY) Toto Supartono1, Iing Nasihin2, Dede Kosasih3 1,2,3Fakultas Kehutanan, Universitas Kuningan Jl. Tjut Nyak Dhien, No.36-A, Cijoho, Kabupaten Kuningan 1Email: toto.supartono@uniku.ac.id keyword: empowerment, Gunung Ciremai, national parks, nurseries. ABSTRACT Nurseries are one of the important means in ecosystem restoration activities because they can act as a source of seed propagation. The purpose of this community service was to build a nursery in the Utilization Zone of Gunung Ciremai National Park in which there are community groups that manage tourism objects. Kompepar Pasir Batang has become a partner in community service activities. The method used was the delivery of materials and field implementation. Nurseries can be built as expected. The results of the activity show that the community gives a positive response to community service activities carried out by educational institutions. keyword: empowerment, Gunung Ciremai, national parks, nurseries. 602 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 PENDAHULUAN Kelompok masyarakat yang akan dijadikan mitra pada kegiatan ini adalah Kelompok Penggerak Pariwisata (Kompepar) Pasir Batang. Kelompok ini sejak tahun 2017 mengelola objek wisata sebagai bumi perkemahan yang berada di Zona Pemanfaatan Taman Nasional Gunung Ciremai. Meskipun dilakukan di kawasan hutan negara, kegiatan pengelolaan wisata yang dilakukan kelompok ini adalah legal karena mendapatkan ijin dari Kementrian Lingkungan Hidup dan Kehutanan (KLHK) melalui Balai Taman Nasional Gunung Ciremai (BTNGC). Secara administrasi pemerintahan, kelompok ini berada dalam Desa Karangsari, Kecamatan Darma, Kabupaten Kuningan. Gunung Ciremai pada mulanya berupa hutan produksi dan menjadi taman nasional semenjak tahun 2004 (Kementrian Kehutanan 2004). Areal yang dikelola oleh mitra berupa lahan yang tertutup oleh tegakan pinus (Pinus merkusii). Tegakan pinus sudah ada sejak areal ini masih dikelola oleh Perum Perhutani dan ketika itu berfungsi sebagai hutan produksi. Kawasan konservasi idealnya tertutup oleh ekosistem atau tegakan yang bersifat alami, bukan berupa tegakan pinus. Akan tetapi, penebangan pohon dalam rangka penggantian tegakan yang alami di dalam kawasan konservasi belum ada aturannya. Oleh karena itu, salah satu upaya yang lebih memungkinkan adalah dengan melakukan kegiatan pengkayaan dengan menanam jenis- jenis lokal di sela-sela pohon pinus. Selain itu, berbatasan dengan areal ijin kegiatan wisata, terdapat tutupan lahan yang masih didominasi oleh semak belukar. Terkait dengan hal tersebut, mitra juga dituntut untuk berpartisipasi dalam memulihkan ekosistem yang didominasi semak belukar. Salah satu upaya yang dapat dilakukan untuk mempercepat pemulihan kondisi ekosistem adalah melakukan kegiatan penanaman. Guna melakukan kegiatan pengkayaan dan penanaman pada ekosistem yang perlu dipulihkan, mitra saat ini masih memiliki beberapa keterbatasan di antaranya persemaian. Dengan kata lain, mitra belum memiliki persemaian yang dapat menunjang kegiatan pengkayaan dan pemuilihan ekosistem. Padahal, keberadaan persemaian sangat diperlukan untuk pembiakan dan perbanyakan anakan dari jenis-jenis yang sesuai untuk kegiatan pengkayaan dan pemulihan ekosistem. Selain itu, mitra juga masih memiliki keterbatasan 603 pengetahuan dalam pembuatan persemaian. Oleh karena itu, pendampingan pembuatan persemaian sangat diperlukan. Tujuan dari kegiatan pengabdian ini adalah melakukan pemberdayaan mitra dalam pembuatan persemaian di dalam kawasan konservasi Taman Nasional Gunung Ciremai, yang masuk ke dalam wilayah adminsitrasi Desa Karangsari, Kecamatan Darma, Kabupaten Kuningan. METODE PELAKSANAAN Bagian Kegiatan dilakukan di salah satu Zona Pemanfaatan; tepatnya di blok Pasir Batang Taman Nasional Gunung Ciremai. Tutupan lahan di sekitar lokasi kegiatan berupa tegakan hutan pinus yang sudah cukup tua (Supartono dan Yudayana 2019) serta semak belukar (Supartono et al. 2019) yang sudah bercampur dengan tumbuhan kaliandra (Supartono et al. 2018). Kegiatan dilakukan pada bulan Oktober 2020. Kegiatan diawali dengan konsolidasi dan sosialisasi dengan mitra dan diakhiri dengan penyusunan laporan. Sebagaimana telah disebutkan di atas, salah satu permasalahan dalam pengkayaan dan pemulihan ekosistem di sekitar wilayah pengelolaan mitra adalah belum tersedianya persemaian. Oleh karena itu, solusi yang telah ditawarkan adalah pembuatan persemaian. Selain dalam bentuk diagram alir (Gambar 1), langkah-langkah yang dilakukan dalam kegiatan ini secara lebih rinci adalah sebagai berikut: 1. Sosialisasi Sosialisasi telah dilakukan pada tahap pertama kegiatan. Sosialisasi bertujuan agar semua anggota mitra mengetahui mengenai kegiatan yang akan dilakukan. Dengan harapan, semua anggota dapat terlibat dalam kegiatan ini. Berdasarkan informasi dari ketua kelompok, tingkat keterlibatan anggota mitra selama ini masih cukup rendah. Bahkan terdapat anggota kelompok yang dapat dikatakan tidak aktif dalam kegiatan pengelolaan wisata alam. 2. Penyampaian Teknis Penyampaian teknis dilakukan setelah selesai pelaksanaan sosialisasi. Tujuan penyampaian teknis adalah agar semua anggota mitra benar-benar faham apa yang harus dilakukan dalam pembuatan persemaian ini. Dalam kegiatan ini, materi yang disampaikan tidak hanya terkait teknis pembuatan persemaian, melainkan juga manfaat dari keberadaan persemaian bagi pengkayaan dan pemulihan eksositem serta keuntungan-keuntungan ketika ekosistem di sekitar sudah pulih. 604 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 4. Pembangunan Bedeng Persemaian Pembangunan bedeng persemaian dilakukan oleh mitra dengan arahan dari fakultas kehutanan. Ukuran bedeng persemaian menyesuaikan dengan kondisi di lapangan. Gambar 1. Diagram Alir Pelaksanaan Pembuatan Bedeng Persemaian Sosialisasi Kegiatan Penyampaian Teknis Pengadaan Alat dan Bahan Pembangunan Bedeng Sosialisasi Kegiatan Penyampaian Teknis Pengadaan Alat dan Bahan Pembangunan Bedeng Gambar 1. Diagram Alir Pelaksanaan Pembuatan Bedeng Persemaian 3. Pengadaan Alat dan Bahan Pembiayaan untuk pengadaan bahan dan alat dibantu oleh lembaga, dalam hal ini Fakultas Kehutanan Universitas Kuningan. Akan tetapi, mengingat kegiatan ini merupakan pemberdayaan masyarakat, maka teknis pengadaan bahan dan alat dilakukan oleh mitra. Hal ini bertujuan agar mitra memperoleh pengalaman dalam pengadaan bahan dan alat dan manajemen biaya. Partisipasi Mitra dalam Pelaksanaan Program p g Kegiatan ini melibatkan Kelompok Penggiat Pariwisata Bumi Perkemahan Pasir Batang karena salah satu harapan dari kegiatan ini adalah peningkatan partisipasi masyarakat dalam pemanfaatan limbah-limbah pertanian terutama sayuran. Bentuk keterlibatan kelompok masyarakat dalam kegiatan ini adalah: pembuatan alat komposer/fermenter, pengumpulan bahan baku, pencacahan, dan pencampuran cairan Partisipasi Mitra dalam Pelaksanaan Program Kegiatan ini telah melibatkan anggota Kompepar Pasir Batang, Karangsari. Hal tersebut karena salah satu target dan harapan dari kegiatan ini adalah meningkatnya partisipasi masyarakat dalam pemulihan kualitas ekosistem TNGC yang saat ini pada beberapa titik masih berupa semak belukar, melalui permbuatan persemaian sebagai tempat pembiakan bibit rehabilitasi, khsususnya blok hutan Pasir Batang. Bentuk keterlibatan Kompepar dalam kegiatan ini adalah: identifikasi lokasi, penyiapan tempat, penyiapan bahan material, dan pembuatan bedeng persemaian. 605 1). Sosialisasi ) Langkah pertama yang telah dilakukan dalam pelaksanaan kegiatan pengabdian kepada masyarakat ini adalah melakukan sosialisasi (Gambar 2). Kegiatan sosialisasi dilakukan bersamaan dengan tema pengabdian yang lain yang dilakukan oleh tim yang berbeda. Hal ini dilakukan dengan tujuan supaya semua anggota memperoleh informasi dan penjelasan yang komprenhensif dari beberapa aspek kehutanan dan lingkungan. Gambar 2. Sosialisasi Kegiatan Pengabdian Kepada Masyarakat Kepada Anggota Kompepar Pasir Batang Serta Karangtaruna Desa Karangsari, Kecamatan Darma, Kabupaten Kuningan Gambar 2. Sosialisasi Kegiatan Pengabdian Kepada Masyarakat Kepada Anggota Kompepar Pasir Batang Serta Karangtaruna Desa Karangsari, Kecamatan Darma, Kabupaten Kuningan Materi yang disampaikan pada kegiatan ini mencakup: maksud dilaksanakannya kegiatan, pentingnya peranan persemaian dan teknis pembuatan persemaian. Materi lainnya yang disampaikan adalah pentingnya memulihkan ekosistem dan penjelasang tentang beberapa dampak negatif ketika ekosistem hutan mengalami kerusakan. Pada kesempatan tersebut juga disampaikan korelasi antara kerusakan hutan dengan penyebaran 606 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 virus yang didasarkan pada publikasi hasil penelitian-penelitian yang sudah ada (seperti hasil publikasi Afelt et al. 2018). Penyampaian informasi ini bertujuan untuk menggugah dan meningkatkan kesadaran pentingnya menjaga kelestarian hutan termasuk mendukung kegiatan pembuatan persemaian di lokasi kegiatan. virus yang didasarkan pada publikasi hasil penelitian-penelitian yang sudah ada (seperti hasil publikasi Afelt et al. 2018). Penyampaian informasi ini bertujuan untuk menggugah dan meningkatkan kesadaran pentingnya menjaga kelestarian hutan termasuk mendukung kegiatan pembuatan persemaian di lokasi kegiatan. 2). Penyampaian Teknis Pembuatan Persemaian Penyampaian teknik pembuatan persemaian secara rinci juga dilakukan sesaat sebelum dilakukan pembuatan persemaian (Gambar 3). Pada kesempatan ini juga disampaikan rencana ke depan jenis-jenis yang akan disemaikan; yaitu jenis-jenis pohon setempat atau pohon lokal. Hal tersebut mengingat kegiatan pemulihan ekosistem akan dilakukan di dalam kawasan konservasi. Sesuai dengan aturan yang berlaku, jenis yang boleh ditanam di dalam kawasan konservasi adalah jenis setempat dan tidak boleh menanam jenis-jenis asing, terlebih lagi jenis-jenis invasif; invasif alien species (IAS). Gambar 3. Penyampaian Teknik Pembuatan Persemaian kepada Anggota Kompepar Bumi Perkembahan Pasir Batang, Desa Karangsari Gambar 3. Penyampaian Teknik Pembuatan Persemaian kepada Anggota Kompepar Bumi Perkembahan Pasir Batang, Desa Karangsari Materi teknik pembuatan persemaian mencakup ukuran persemaian, ukuran setiap bedeng, kapasitas setiap bedeng, material penutup persemaian. Selain itu, materi yang disampaikan juga adalah terkait dengan bahan yang akan digunakan pada pembangunan persemaian serta pertimbangan/alasan digunakannya tersebut. Prinsip yang dipegang bahwa bahan yang digunakan untuk pembuatan persemaian harus ramah lingkungan. 607 3). Pemilihan dan Penyiapan Lahan ) y p Pada tahap ini, mitra juga diminta masukan atau sarannya terkait dengan penentuan lokasi. Prinsip dari lokasi yang digunakan adalah: tidak ditumbuhi anakan secara alami, tidak ada bekas kegiatan penanaman, memiliki kondisi yang cukup datar agar memudahkan dalam pengerjaannya dan banyak ditumbuhi oleh semak belukar. Selain itu, pertimbangan yang dipakai juga adalah lokasinya cukup dekat dengan sumber air. Hal tersebut dilakukan agar memudahkan dalam penyiraman ketika musim kemarau. Penyiapan lahan dilakukan dengan cara membersihkan calon lokasi persemaian dari berbagai tumbuhan bawah atau semak belukar. Jenis-jenis kaliandra meskipun memiliki ukuran yang cukup besar (setara dengan ukuran pancang dan tiang) juga dibersihkan pada lokasi tersebut. Selanjutnya, lahan diratakan (Gambar 4) supaya memudahkan dalam penyimpanan atau penataan polibag yang akan diisi dengan anakan. Gambar 4. Perataan Calon Lokasi Pembuatan Persemaian Gambar 4. Perataan Calon Lokasi Pembuatan Persemaian 4) Penyiapan Material Persemaian 4) Penyiapan Material Persemaian Pembangunan persemaian berusaha semaksimal mungkin menggunakan bahan-bahan yang ada dalam kawasan dengan tetap memperhatikan aspek kelestariannya. Tiang-tiang untuk penutup persemaian menggunakan batang kaliandra (Gambar 5). Mengingat kaliandra merupakan jenis invasif (Mustika 2012; Sunaryo et al. 2012), maka pemanfaatan kaliandra dalam pembuatan persemaian telah membantu dalam mengurangi keberadaan kaliandra di lokasi kegiatan. Jenis lainnya yang digunakan dan berasal dari dalam kawasan adalah bambu, yang kebetulan letak rumpunnya relatif berdekatan dengan lokasi kegiatan. Selain bersama dengan anggota Kompepar, kegiatan ini juga melibatkan para mahasiswa. 608 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 Hal ini bertujuan untuk membiasakan para mahasiswa dalam bersosialisasi dengan masyarakat dan menumbuhkan kepedulian kepada masyarakat. Gambar 5. Pengambilan Batang Kaliandra Sebagai Tiang Penutup/Naungan Persemaian Gambar 5. Pengambilan Batang Kaliandra Sebagai Tiang Penutup/Naungan Persemaian 6). Persemaian yang Sudah Jadi 6). Persemaian yang Sudah Jadi Guna memperoleh pertumbuhan anakan yang optimal di persemaian, arah bedengan juga sebenarnya harus menjadi pertimbangan. Arah bedengan idealnya menghadap Utara – Selatan, agar sinar matahari bisa masuk secara merata (Barkah 2009). Akan tetapi, arah bedengan pada kegiatan ini adalah Timur-Barat. Hal tersebut karena menyesuaikan dengan kondisi lapangan yang ada, di mana arah yang memungkinkan adalah Timur–Barat. Ukuran persemaian yang telah dibuat adalah 11 m x 3 m. Jumlah bedeng yang ada dalam persemaan sebanyak 5 buah dengan ukuran setiap bedeng adalah 3 m x 1 m. Pembatas bedeng menggunakan bambu. Tiang penutup bedeng menggunakan batang kaliandra dengan penutup berupa paranet. Kapasitas setiap bedengan adalah 300 polybag. Dengan demikian, kapasitas persemaian adalah 1500 polybag (Gambar 7). Gambar 7. Persemaian yang Sudah Selesai dengan Kapasitas 1500 polybag/bibit Gambar 7. Persemaian yang Sudah Selesai dengan Kapasitas 1500 polybag/bibit Ketika pembangunan persemaian sudah selesai, selanjutnya terdapat beberapa hal yang harus mendapatkan perhatian agar memperoleh bibit yang bermutu. Hal-hal tersebut adalah a) Pengadaan benih: sumber benih, penanganan benih, dan perlakuan benih; b) Pengadaan dan pengolahan media: perlakuan media, jenis media, dan kesuburan media; c) Pemeliharaan bibit: penyiraman, pemberian naungan, penyiangan bibit, pemupukan, dan perlindungan hama penyakit; dan d) Pengangkutan/transportasi bibit (Barkah 2009). 5). Pembangunan Persemaian g Pada saat semua material yang dibutuhkan sudah tersedia, langkah selanjutnya adalah pembangunan persemaian (Gambar 6). Kegiatan tersebut mencakup pembuatan bedengan dan pemasangan tiang dan penyangga untuk penutup/naungan persemaian. Sebagaimana telah disebutkan di atas, tiang penyangga menggunakan batang kaliandra. Alat untuk penahan naungan menggunakan bambu. Sementara itu, naungan menggunakan paranet dengan intensitas cahaya 50%. Tidak hanya ketika menyiapkan material, pembangunan persemaian juga telah melibatkan mahasiswa. Tujuan utamanya adalah memberikan pengalaman lapangan kepada mahasiswa tentang bagaimana cara membuat persemaian. 609 Gambar 6. Pembangunan Persemaian dengan Menggunakan Bahan-Bahan yang Ada di Sekitar Lokasi Kegiatan UCAPAN TERIMA KASIH Ucapan terimakasih disampaikan kepada 1) Universitas Kuningan yang sudah membiayai kegiatan pengabdian kepada masyarakat melalui hibah internal, 2) Kelompok Penggerak Parawisata Pasir Batang Taman Nasional Gunung Ciremai yang telah menjadi mitra pada kegiatan ini, 3) Balai Taman Nasional Gunung Ciremai yang sudah memberikan ijin melaksanakan kegiatan di dalam kawasan. 7) Pelajaran yang Diperoleh ) j y g p Selama kegiatan berlangsung anggota Kompepar memberikan respon positif bahkan mereka sangat senang dengan adanya kegiatan ini. Hal tersebut karena mereka telah 610 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 Jurnal Berdaya Mandiri Vol. 3 No. 2 Tahun 2021 E-ISSN: 2685-8398 memperoleh pengetahuan dari lembaga pendidikan yang berada di Kabupaten Kuningan. Anggota Kompepar lebih menguasai kondisi lingkungan sekitar sehingga sangat tepat bila beberapa kegiatan yang dilakukan oleh lembaga pendidikan di wilayah tersebut melibatkan anggota Kompepar. Oleh karena itu, pelibatan anggota Kompepar juga dapat dijadikan untuk penggalian pengalaman lapangan oleh lembaga pendidikan tinggi. KESIMPULAN Kegiatan pemberdayaan masyarakat melalui pembangunan persemaian telah mendapatkan respon positif dari mitra. Kedua belah pihak telah sama-sama mendapatkan manfaat: pihak dari lembaga pendidikan dapat berbagi pengetahuan dan memperoleh pengalaman lapangan dari mitra; demikian juga mitra telah mendapatkan pengetahuan dari pihak lembaga pendidikan. Pembangunan persemaian telah menjadi sarana untuk berbagi ilmu pengetahuan dan meningkatkan komunikasi antara masyarakat dan lembaga pendidikan. REKOMENDASI Rekomendasi yang dapat disampaikan dari kegiatan ini adalah lembaga pendidikan tetap melibatkan masyarakat melalui Anggota Kompepar dalam melakukan pengelolaan persemaian selanjutnya. Afelt, A., Frutos, R., & Devaux, C. (2018). Bats, Coronaviruses, and Deforestation: Toward the Emergence of Novel Infectious Diseases? Front Microbiol, 9, 702. Doi: 10.3389/fmicb.2018.00702. DAFTAR PUSTAKA 611 Barkah, B.S. (2009). Panduan Pembangunan dan Pengelolaan Persemaian Desa Program Rehabilitasi Hutan Rawa Gambut Berbasis Masyarakat di Areal MRPP Kabupaten Musi Banyuasin. Palembang: German Technical Cooperation. [Kemenhut] Kementrian Kehutanan. (2004). Keputusan Menteri Kehutanan Nomor : 42/Menhut-II/2004 tentang Penunjukan Kawasan Lindung Kelompok Hutan Gunung Ciremai menjadi Taman Nasional. Jakarta: Kementrian Kehutanan. Mustika, E. (2012). Analisis Distribusi Spesies Invasif Kaliandra (Calliandra calothyrsus Meissn.) di Taman Hutan Raya Bung Hatta Sumatera Barat. Masters thesis, Universitas Andalas. Sunaryo, Uji, T., & Tihurua, E.F. (2012). Komposisi Jenis dan Potensi Ancaman Tumbuhan Asing Invasif di Taman Nasional Gunung Halimun-Salak, Jawa Barat. Berita Biologi, 11(2), 231 – 239. Supartono, T., Adhya, I., & Yudayana, B. (2018). Soil Seed Bank Germination in Pine Forests and Shrubs, In Gunung Ciremai National Park. Journal of Forestry and Environment, 02, 18-21. Supartono, T., & Yudayana, B. (2019). Partisipasi Masyarakat dalam Peningkatan Pertumbuhan Permudaan Alami Di Bumi Perkemahan Pasir Batang Taman Nasional Gunung Ciremai Desa Karangsari, Kecamatan Darma, Kuningan, Jawa Barat. Empowerment, 02(01), 38-45. Supartono, T., Nasihin, I., Yudhayana, B., Ahsari, O.T., Dahrun, I., & Triliantho, S. (2019). Pemberdayaan Kelompok Penggiat Pariwisata Bumi Perkemahan Pasir Batang Dalam Pemanfaatan Limbah Sayuran Sebagai Pupuk Organik Cair, Desa Karangsari, Kecamatan Darma, Kabupaten Kuningan. Empowerment, 02(02), 74-82. 612	45,696
https://openalex.org/W3162721118	OpenAlex	Open Science	CC-By	2,021	Book review: &lt;i&gt;From Crust to Core – A Chronicle of Deep Carbon Science&lt;/i&gt; by Simon Mitton	Mohamed Ali Bouhifd	English	Spoken	1,030	1,555	Mohamed Ali Bouhifd Laboratoire Magmas et Volcans, CNRS & Université Clermont Auvergne Campus universitaire des Cézeaux, 6 Avenue Blaise Pascal, 63170 Aubière, France Mohamed Ali Bouhifd Laboratoire Magmas et Volcans, CNRS & Université Clermont Auvergne Campus universitaire des Cézeaux, 6 Avenue Blaise Pascal, 63170 Aubière, France Correspondence: Mohamed Ali Bouhifd (ali.bouhifd@uca.fr) Published: 11 May 2021 Correspondence: Mohamed Ali Bouhifd (ali.bouhifd@uca.fr) Mitton, S.: From Crust to Core – A Chronicle of Deep Carbon Science, Cambridge University Press, Univer- sity Printing House, Cambridge CB2 8BS, UK, 382 pp., ISBN: 9781316997475, EUR 44.99, 2020. teriors, but also the genesis of a new interdisciplinary ﬁeld: oceanography. In chapters 12–14 the focus is on the cycles, reservoirs and the ﬂuxes of carbon between the main Earth reservoirs and also on the history of high-pressure physics and chemistry of carbon-bearing minerals under the Earth’s mantle conditions. In the ﬁnal chapter, research on deep life is told from the studies by the pioneer Alexander von Hum- boldt to our days. For every chapter, the author provides a very interesting list of references reporting some remarkable discoveries that changed our understanding of Earth-system science as a whole. Carbon is the fourth most abundant element in the solar sys- tem. Its presence signiﬁcantly affects chemical and physical processes in the Earth’s atmosphere, mantle and core. The importance of carbon also lies in the fact that it has been es- sential for the origin and evolution of life, that it is involved in an important fraction of the energy we use, and that it is as- sumed to play a fundamental role in the Earth’s climate sys- tem. In From Crust to Core – A Chronicle of Deep Carbon Science, Simon Mitton explores several centuries of philo- sophical and scientiﬁc inquiry on the nature of the Earth’s interior, as based on the stories of over 100 scientists, and sheds light on the particular roles of deep carbon. The author is a life fellow at St Edmund’s College at University of Cam- bridge. His current research discipline is history of science, and his ﬁeld of expertise is the history of astronomy and plan- etary science in the nineteenth and twentieth centuries. He is a member of the Deep Carbon Observatory (Carnegie In- stitution for Science, Washington, DC) network of scientists whose mission is to understand better the element carbon in Earth. Hist. Geo Space Sci., 12, 95–96, 2021 https://doi.org/10.5194/hgss-12-95-2021 © Author(s) 2021. This work is distributed under the Creative Commons Attribution 4.0 License. Book review Published by Copernicus Publications. Mohamed Ali Bouhifd Laboratoire Magmas et Volcans, CNRS & Université Clermont Auvergne Campus universitaire des Cézeaux, 6 Avenue Blaise Pascal, 63170 Aubière, France In summary, the book introduces the history of Earth’s car- bon all the way from its synthesis in the ﬁrst generation of stars in our universe, to its incorporation in the solar nebula, where the Sun and planets formed nearly 4.5 billion years ago, to its ﬁnal delivery to Earth with chondrites. The book takes the reader on a captivating trip with an excellent nar- rative style. Many discoveries, which led to enormous ad- vances in broadening our understanding of the history of science in general, are told, for example, the discovery by Hans Bethe (Nobel Prize in Physics in 1967), who made a breakthrough regarding the synthesis of elements in stars, by showing the carbon role in the astrophysical conversion of mass into energy, validating Einstein’s theory, E = mc2. The book also includes the history of the discovery of ra- dioactivity, a process for which Becquerel, Pierre and Marie Curie were awarded the Noble Prize in Physics in 1903. This discovery would have a huge impact on the Earth’s internal dynamics over its 4.5 billion years of history owing to the important amount of thermal energy produced by radioactive elements present within the Earth. The discovery of radioac- tivity provided a novel and robust way to measure the age of rock samples that made most other geological dating meth- ods obsolete. The book includes the fascinating history of the Composed of 15 chapters, the book starts with why car- bon in Earth matters and follows with the origin of carbon and its delivery to Earth. In chapters 4–7, the author presents scientiﬁc adventures behind the determination of the age and of the physical and chemical interior of the Earth. In chap- ters 8–10, the reader will meet a number of remarkable sci- entists whose discoveries were to have major implications for understanding not only the dynamics of the Earth’s in- M. A. Bouhifd: Book review: From Crust to Core – A Chronicle of Deep Carbon Science 96 theory of plate tectonics and a history of research on deep di- amonds, which can keep the chemical message of their for- mation billions or millions of years ago. It also includes the history of the pioneering research on deep life during the last half a century of extraordinary progress on questions about the limits of life on Earth and the nature of deep life. Hist. Geo Space Sci., 12, 95–96, 2021 Mohamed Ali Bouhifd Laboratoire Magmas et Volcans, CNRS & Université Clermont Auvergne Campus universitaire des Cézeaux, 6 Avenue Blaise Pascal, 63170 Aubière, France In a ﬁ- nal note, the author reaches the philosophical conclusion that there is a great deal still to be discovered about the microbial environment of Earth’s interiors that is highly relevant to the search for life elsewhere. Therefore, it seems entirely plausi- ble that, right beneath our feet, deep carbon science could yet uncover important clues on the origin of life in the universe. This is an excellent book for everyone interested in know- ing the stories behind the key discoveries in deep carbon sci- ence that includes (but not restricted to) the Big Bang, origin of carbon in the universe, formation of rocky planets, dia- monds in the Earth’s mantle and the signs of deep life. Addi- tionally, From Crust to Core – A Chronicle of Deep Carbon Science is well affordable for all “amateurs” and also profes- sionals who are looking for the fascinating story of the evo- lution of Earth system science in general and deep carbon science in particular. https://doi.org/10.5194/hgss-12-95-2021	41,946
https://openalex.org/W2994829992	OpenAlex	Open Science	CC-By	2,019	Endogenous Bufadienolides, Fibrosis and Preeclampsia	Vitaly A. Reznik	English	Spoken	5,942	11,531	1. Introduction Hypertension is the most common medical condition encountered during pregnancy, with a frequency of ap- proximately 6–8% [1]. Te hypertensive disorders of pregnancy cover a spectrum of conditions, including preeclampsia-eclampsia, gestational hypertension, and chronic hypertension. Working Group on High Blood Pressure in Pregnancy deﬁnes hypertension in pregnancy as two blood pressure measurements of ≥140/90 mmHg measured≥six hours apart [1]. Despite ongoing prevention and repeated attempts to select a therapy, it was not possible to reduce the frequency of preeclampsia (PE). Te diagnosis of PE is made by the combination of arterial hypertension (systolic blood pressure ≥140 mmHg and(or) diastolic blood pressure ≥90 mmHg [2]), proteinuria (presence of protein in the urine ≥0.3 g/l in the daily sample (24 h) or in two samples taken at intervals of 6 h; using the test strip (protein in the urine) [2], the indicator (0.3 g/l [3])), and generalized edema. Peripheral edema is observed in 70–80% of cases with physiological pregnancy, therefore, cannot be considered as a diagnostic criterion for PE [4]. Vitaliy A. Reznik,1 Vladimir A. Kashkin,2,3 Natalia I. Agalakova ,3 C. David Adair,4 and Alexei Y. Bagrov 3 1Department of Obstetrics and Gynecology, School of Pediatric Medicine, St. Petersburg 194353, Russia 2Valdman Institute of Pharmacology, Pavlov Medical University, St. Petersburg 197022, Russia 3Sechenov Institute of Evolutionary Physiology and Biochemistry, 44 Torez Prospect, St. Petersburg 194223, Russia 4Department of Obstetrics and Gynecology, University of Tennessee, Chattanooga, TN, USA Correspondence should be addressed to Alexei Y. Bagrov; aybagrov@gmail.com Received 11 June 2019; Accepted 21 November 2019; Published 12 December 2019 Academic Editor: Syed Wamique Yusuf Copyright © 2019 Vitaliy A. Reznik et al. Tis 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. Frequency of preeclampsia has no tendency to decrease, and it still takes the leading position in the structure of maternal mortality and morbidity worldwide. In this review, we present the “ﬁbrotic concept” of the etiology and pathogenesis of preeclampsia which involves system consisting of Na/K-ATPase and its endogenous ligands including marinobufagenin. New therapy of preeclampsia includes modulation of the Na/K-ATPase system by immunoneutralization of the marinobufagenin and use of mineralocorticoid antagonists which are capable to impair marinobufagenin-Na/K-ATPase interactions. Te mechanisms implicated in the pathophysiology of PE include oxidative stress, placental steroid formation, and insulin resistance [5, 6]. Te endpoint of this complex in- terplay between maternal and placental mechanisms is a maternal multisystem disorder, characterized by hyperten- sion, proteinuria, and multiorgan dysfunction. During PE, the placental spiral arteries fail to lose their elastic lamellae ultimately leading to decreased placental perfusion [7]. Placental hypoxia is frequently viewed as an early trigger of placental production of soluble factors resulting in endo- thelial dysfunction [8], which may play the main role in the pathogenesis of PE. Hindawi Cardiology Research and Practice Volume 2019, Article ID 5019287, 7 pages https://doi.org/10.1155/2019/5019287 Hindawi Cardiology Research and Practice Volume 2019, Article ID 5019287, 7 pages https://doi.org/10.1155/2019/5019287 Hindawi 2. Fibrosis and Preeclampsia Te ﬁrst observation suggesting that ﬁbrosis can underlie the development of PE is related to the works of Tenney and Montgomery in 1935–1936. It has been shown that collagen ﬁbers in the villous stroma are ﬁrst visible on light mi- croscopy as early as the 2nd month of gestation. Te authors found that the collagen content of the villi showed no ap- preciable increase between midpregnancy and midterm [9]. In 1968, Fox conﬁrmed empirically the results of these works 2 Cardiology Research and Practice PE. In the “pathophysiologically relevant” range of con- centrations (1–3 nmol/L), in vitro, MBG exhibits a 25% inhibition of NKA in the human vascular sarcolemma and induces a contractile response in the isolated rings of human mesenteric arteries [19]. Tus, the in vivo levels of MBG observed in patients with PE in our previous and present studies are capable of enhancing vascular tone and sub- stantially inhibiting NKA. and suggested that placental ﬁbrosis is not associated with any excess of fetal hypoxia complications. It is suggested that villous ﬁbrosis is a result of a reduced fetal villous blood ﬂow; such a reduction may be due, as in toxemia or diabetes, to obliterating endarteritis changes in the fetal stem arteries or, as in prolonged pregnancies, to a constriction of villous capillaries [9]. p [ ] In later works, it was shown that women who have experienced PE are at increased risk of hypertension, cor- onary artery disease, and fatal stroke. Its eﬀects do not end with childbirth and is associated with a higher cardiovas- cular risk lifelong. Tis increased risk could be partly explained by left ventricular systolic and diastolic impair- ment, arterial stiﬀening, and endothelial dysfunction which have been described during and after PE. Tese phenomena, which inﬂuence each other, are related to myocardial and vascular remodeling, a process leading to loss of function and ﬁbrotic tissue deposition [10–12]. In early studies, it was suggested that the modiﬁed placental production of vaso- active substances appears to underlie the pathological mechanism of hypertension and vascular dysfunction in PE [13]. Over the past 20 years, it has been shown that en- dogenous digitalis-like factors or cardiotonic steroids (CS) of the Na/K-ATPase (NKA) are also among these substances, and elevated plasma levels of CS were detected in patients with PE [14–16]. 2. Fibrosis and Preeclampsia Te mechanism of PE includes remodeling of uterine spiral arteries and making them ﬁbrotic by a variety of vascular factors including cardiotonic steroids [20, 21]. Physiological function of CS is regulation of sodium ex- cretion by means of inhibition of NKA in the epithelial cells of renal tubuli, i.e., an adaptive response [20]. Under pathological conditions, for example, in patients with PE, side eﬀect of this primarily adaptive response becomes es- sential, and CS cause inhibition of sodium pumps in tissues including vessels [22]. Recent studies demonstrated that in addition to vasoconstrictor eﬀect of CS, these compounds directly aﬀect intracellular signaling resulting in the loss of vascular elasticity and vascular ﬁbrosis [23]. Low concen- trations of CS acting on PKC delta induce deactivation of Fli-1 and cause expression of collagen-1, which is a key factor of ﬁbrosis in blood vessels, myocardium, and kidney [23, 24]. Recent studies demonstrate the clinical importance of ﬁbrosis in the cardiovascular system and placenta [25, 26], and moreover, suggest that blood vessels exposed to PE environment exhibit greater responsiveness to injury despite resolution of PE [27]. Te general scheme of the develop- ment of MBG-induced ﬁbrosis is presented in Figure 2. Recently, it has been demonstrated that nanomolar concentrations of bufadienolide CS, marinobufagenin (MBG) (Figure 1) stimulate the synthesis of collagen and induce ﬁbrosis in cardiovascular tissues and in the kidney [17]. One of the mechanisms underlying the proﬁbrotic eﬀect of MBG is EGFR-Src-dependent inhibition of Fli-1, a nuclear transcription factor and a negative collagen-1 reg- ulator, causing its reduction which resulted in activation of collagen-1 [15]. During the experiments, it was shown that CS (MBG, ouabain, and digoxin) cause an increase in the number of embedded proline residues and also increase collagen synthesis in the ﬁbroblast culture, which was evi- dence of the presence of signaling function of the Na/K- ATPase-CS complex with activation of Src and MAPK [17, 18]. In vivo, the administration of MBG in a concen- tration that is observed in renal failure caused the devel- opment of cardiac ﬁbrosis with activation of the signaling pathway mediated by NKA, which was conﬁrmed by an increase in Src and phosphorylation of a mitogen-activated protein kinase (MAPK) in the myocardium [17, 18]. 3. Treatment of Preeclampsia For millennia, the venomous secretions of amphibians, which contain a large number of bufadienolide derivatives, are widely used in the traditional medicine of diﬀerent civilizations. One of the most well-known examples from the traditional Chinese medicine is Sen-So (or Ch`an Su), which was used for the treatment of heart failure [28]. Being highly active substances, bufadienolides often caused toxic eﬀects. Human fatal poisonings from toad venom have been re- ported in the many countries by the venoms from the Colorado River toad (Bufo alvarius), the cane toad (Bufo marinus), and the Chinese toad (Bufo gargarizans) [29]. Over the past 20 years, the digoxin-speciﬁc Fab fragments (Digibind) have been successfully used in the poisoning of toad venom, and it has shown their eﬀectiveness in animals [30] and humans [31]. It is well known also that in patients at the ﬁnal stage of renal failure [32] or PE [33], as well as in Dahl-S (salt sensitive) rats with salt load [34, 35], the erythrocyte NKA is inhibited, which can be reversed in- cubation, ex vivo with antibodies to MBG or with Digibind, and bovine digoxin antibodies that interact with CS [33, 36]. Our observations conﬁrmed and extended previously obtained ﬁndings demonstrating substantial elevations of MBG levels in PE. In our previous study, in patients with severe PE (161/104 mm Hg), plasma MBG concentration comprised averaged 2.63 nmol/L. Later, it was shown that in subjects with mild PE and lower arterial pressure, the mean level of MBG in plasma was 1.68 nmol/L [19]. Since plasma concentrations of MBG in subjects with noncomplicated pregnancy in these previous studies were similar and comprised 0.80 nmol/L and 0.63 nmol/L, respectively, the levels of MBG may be one of the markers of severity of PE. Our ﬁndings also indicate that MBG may not only be a marker but also a factor contributing to the pathogenesis of 4. Monoclonal Antibody Our labo- ratory demonstrated that in PE plasma levels of MBG in- crease and that ex vivo anti-MBG antibody reverses PE- induced inhibition of erythrocyte NKA [19, 33]. In pregnant NaCl-supplemented rats, increased MBG levels accompa- nied the development of symptoms of PE including eleva- tion of arterial pressure, proteinuria, and reduction of fetal weight and size [33, 42]. In this model, in vivo immuno- neutralization of MBG with poly- and monoclonal anti- bodies produced antihypertensive eﬀect associated with the increase in vascular NKA activity [33, 42]. Considering that, (i) MBG stimulates synthesis of collagen, (ii) ﬁbrosis in placenta and umbilical arteries in PE patients is accompa- nied by elevated placental MBG and a dramatic decrease in Fli-1 level [43], and (iii) PE is associated with vascular stiﬀening [44, 45]; it has been showed that MBG is one of the factors, implicated in pathogenesis of PE via induction of vasoconstriction and vascular ﬁbrosis [41]. In the recent study, it has been shown that the placental levels of Fli-1 were dramatically lower, while collagen-1 levels were higher in both placenta and umbilical arteries in PreE samples compared to the tissues from control noncomplicated pregnancy. Te subsequent treatment of the umbilical artery explants from PE patients with anti-MBG mAb was ac- companied by a signiﬁcant decrease in collagen-1 abundance [43]. Mineralocorticoid antagonists are known to reverse myocardial ﬁbrosis [53]. It has been established that min- eralocorticoid antagonists decrease blood pressure in rat hypertension models and suppress cardiac ﬁbrosis, in which levels of CS including MBG are elevated [48, 49]. Tus, an interaction between Na/K-ATPase and MBG could be a target for aldosterone antagonists [53]. Considering the fact that MBG induces ﬁbrosis in the cardiovascular system through the Fli-1-dependent mechanism, it was investigated the eﬀects of spironolactone and its main metabolite, can- renone, for ﬁbrosis in a series of experiments in vitro and in vivo. It has been shown that both spironolactone and canrenone weaken MBG-induced increase in collagen synthesis by myocardial ﬁbroblasts, which was conﬁrmed in vivo by a distinct decrease in cardiac ﬁbrosis caused by experimental renal failure as a result of treatment with spironolactone [53]. As shown earlier, canrenone and spi- ronolactone act as competing inhibitors of CS and NKA [50, 53–55]. 4. Monoclonal Antibody Considering these, it can be assumed that short- term treatment with low doses of spironolactone or can- renone can be a very eﬀective method of treating ﬁbrotic manifestations of PE. Furthermore, in comparative study of the eﬃcacy of speciﬁc antibodies with antidigoxin (DigiFab) and mono- clonal antibody against MBG (3E9), it was shown that monoclonal antibodies restore the activity of PE-inhibited NKA in erythrocytes better than DigiFab, which suggested the involvement of MBG in the pathogenesis of PE [46]. In a recent study, it was shown that the administration of hu- manized monoclonal antibodies (206–208, H1L2) against MBG attenuate MBG-induced downregulated PCNA and upregulated p38 phosphorylation in cytotrophoblast cells, indicating a possible therapeutic action in PE [47]. Tere- fore, immunoneutralization of MBG is a logical step in the treatment of PE. Spironolactone was introduced in 1960, and before 1980, this molecule was used for hypertension and PE [56, 57]. It was noted that when pregnant rats were treated with 40 mg spironolactone from 13 to 21 days of pregnancy, male foetuses showed signs of feminization [58]. Terefore, spi- ronolactone is not advised for humans during pregnancy [59]. Considering that eplerenone has not been associated with adverse eﬀects during pregnancy in animal studies, this drug is likely to be a better choice for use in pregnant women than spironolactone. However, no data on the use of these drugs in human pregnancy exist, and therefore, eplerenone should be used in pregnant women with heart failure only when treatment with other diuretics (such as furosemide) is noneﬀective [60]. 4. Monoclonal Antibody In this study, the umbilical arteries, obtained from pregnant women suﬀering from PE, contained greater amounts of collagen, less Fli-1, and they also showed low sensitivity to the vasorelaxant eﬀect of sodium nitroprusside. Our labo- ratory demonstrated that in PE plasma levels of MBG in- crease and that ex vivo anti-MBG antibody reverses PE- induced inhibition of erythrocyte NKA [19, 33]. In pregnant NaCl-supplemented rats, increased MBG levels accompa- nied the development of symptoms of PE including eleva- tion of arterial pressure, proteinuria, and reduction of fetal weight and size [33, 42]. In this model, in vivo immuno- neutralization of MBG with poly- and monoclonal anti- bodies produced antihypertensive eﬀect associated with the increase in vascular NKA activity [33, 42]. Considering that, (i) MBG stimulates synthesis of collagen, (ii) ﬁbrosis in placenta and umbilical arteries in PE patients is accompa- nied by elevated placental MBG and a dramatic decrease in Fli-1 level [43], and (iii) PE is associated with vascular stiﬀening [44, 45]; it has been showed that MBG is one of the factors, implicated in pathogenesis of PE via induction of vasoconstriction and vascular ﬁbrosis [41]. In the recent study, it has been shown that the placental levels of Fli-1 were dramatically lower, while collagen-1 levels were higher in both placenta and umbilical arteries in PreE samples compared to the tissues from control noncomplicated pregnancy. Te subsequent treatment of the umbilical artery explants from PE patients with anti-MBG mAb was ac- companied by a signiﬁcant decrease in collagen-1 abundance [43]. in the presence of low MBG concentration produced similar phenotype [41]. We hypothesized that aldosterone antag- onists can also reverse MBG-induced vascular ﬁbrosis, in the explants of the thoracic aorta and in the cultured rat vascular smooth muscle cells, and we observed that canrenone suppressed the eﬀect of MBG synthesis of collagen-1 [23]. Tis observation was conﬁrmed by clinical data which showed that in patients with resistant hypertension receiving spironolactone as an addition to the conventional antihy- pertensive therapy, there was a decrease of aortic vascular stiﬀness in parallel with an increase in the erythrocyte NKA [23]. Tis mechanism was also shown on the PE model [41]. In this study, the umbilical arteries, obtained from pregnant women suﬀering from PE, contained greater amounts of collagen, less Fli-1, and they also showed low sensitivity to the vasorelaxant eﬀect of sodium nitroprusside. 4. Monoclonal Antibody Immunoneutralization of the CS in patients with PE is not new. Te similarity of the functional proﬁle (inhibition of NKA) and the structure of cardenolides (digoxin and 3 3 Cardiology Research and Practice Bufo marinus Marinobufagenin Marinobufagenin COOH COOH Membrane Extracellular [K+]o = 4mM [Na+]o = 140mM [CL–, H2CO3]o, etc. = 150mM [K+]o = 140mM [Na+]o = 12mM [CL–, H2CO3]o, etc. = 150mM ATP P 1 2 3 4 5 6 7 8 9 10 TGES COOH Cytoplasmic β-Isoform γ-Isoform (FXYD) α1-Isoform NH2 NH2 NH2 OH H H HO O O O OH H H HO O O O Figure 1: Chemical structures of bufadienolide, marinobufagenin and its pharmacological target, Na/K-ATPase. Figure 1: Chemical structures of bufadienolide, marinobufagenin and its pharmacological target, Na/K-ATP Cytosol Src Na/K-ATPase Nucleus PKC δ PKC δ α1-Isoform Marinobufagenin EGFR P P P P P Fli-1 Fli-1 Col-1 promoter P Membrane Collagen-1 Procollagen-1 Procollagen-1 mRNA Phospholipase C Proteolysis OH H H HO O O O Figure 2: Scheme of the Fli-1-dependent proﬁbrotic pathway activated via binding of marinobufagenin (MBG) to Na/K-ATPase (NKA). Te interaction of MBG with NKA leads to activation/phosphorylation of Src followed by activation/phosphorylation PLC and PKC via scaﬀolding function of NKA. Marinobufagenin α1-Isoform Cytosol Figure 2: Scheme of the Fli-1-dependent proﬁbrotic pathway activated via binding of marinobufagenin (MBG) to Na/K-ATPase (NKA). Te interaction of MBG with NKA leads to activation/phosphorylation of Src followed by activation/phosphorylation PLC and PKC via scaﬀolding function of NKA. reactivity against the bufadienolides and cardenolides [19]. In a rat model of PE, the Digibind treatment produced lowering of the blood pressure and proteinuria [40]. In a clinical study, it was shown that antibody therapy by Digibind prevents a decline in renal function with well tolerated in severe PE by neutralizing CS [37]. ouabain) and bufadienolides (MBG and resibufogenin) allowed clinical use of polyclonal antibodies against digoxin (Digibind) in the treatment of PE [37, 38]. Digibind and DigiFab have been used for many years to treat digoxin in- toxication [39]. Digibind has been shown to induce a decrease in blood pressure in animals with volume-expanded hyper- tension by an interaction between Digibind and an endog- enous digoxin-like factor [39]. Digibind demonstrates cross- Based on the results of previous studies, the involvement of MBG was demonstrated in the development of ﬁbrosis. Cardiology Research and Practice 4 Tis mechanism was also shown on the PE model [41]. 6. Conclusion Currently, it is known that MBG is actively involved in the pathogenesis of the development of PE. Being an endoge- nous ligand of the NKA, MBG via the ionic and signaling pathways participates in the development of placental ﬁ- brosis, which may be the main pathological mechanisms of PE. We suggest that the system of MBG and the NKA is a keystone of PE pathogenesis. Immunoneutralization of MBG, as well as antagonism against binding with the NKA, can be a new and eﬀective direction in the treatment of PreE and further prevention of its complications. In conclusion, our results indicate that in preeclampsia, elevated levels of MBG, via a Fli-1-dependent mechanism, stimulate synthesis of collagen in umbilical arteries, which leads to an im- pairment of vasorelaxation, and MBG-induced Fli-1-de- pendent signaling pathway contributes to vascular stiﬀness in preeclampsia. Recent studies have provided evidence that an excess of signaling molecules released by the placenta to circulation, soluble FMS-like tyrosine kinase-1 (sFlt1) and endoglin (sEng), or decreased levels of placental growth factor (PlGF) and vascular endothelial growth factor A (VEGF-A) play an important role in the pathogenesis of PE. Numerous candidate biomarkers have been proposed for prediction of PE and measurement of maternal circulating factors, such as ratio of sFlt-1/PlGF and reﬂected anti- angiogenic balance that characterizes PE [66]. In our earlier work, in patients with PE levels of sFlt1 and of sEng were elevated together with CS [42], but whether or not both are interconnected remains to be understood. Interestingly, most of the studies of CS and of antiangiogenic balance did not make a distinction between early- and late-onset PE, which also represents an important area for future investigations. [6] F. Herse and B. LaMarca, “Angiotensin II type 1 receptor autoantibody (AT1-AA)-mediated pregnancy hypertension,” American Journal of Reproductive Immunology, vol. 69, no. 4, pp. 413–418, 2013. [7] J. W. Meekins, R. Pijnenborg, M. Hanssens, I. R. McFadyen, and A. van Asshe, “A study of placental bed spiral arteries and trophoblast invasion in normal and severe pre-eclamptic pregnancies,” British Journal of Obstetrics and Gynaecology, vol. 101, no. 8, pp. 669–674, 1994. [8] O. Genbacev, Y. Zhou, J. W. Ludlow, and S. J. Fisher, “Regulation of human placental development by oxygen tension,” Science, vol. 277, no. 5332, pp. 1669–1672, 1997. [9] H. Fox, “Fibrosis of placental villi,” Te Journal of Pathology and Bacteriology, vol. 95, no. 2, pp. 573–579, 1968. [10] A. 6. Conclusion Hausvater, T. Giannone, Y. H. Sandoval et al., “Te as- sociation between preeclampsia and arterial stiﬀness,” Journal of Hypertension, vol. 30, no. 1, pp. 17–33, 2012. [11] L. Bellamy, J.-P. Casas, A. D. Hingorani, and D. J. Williams, “Pre-eclampsia and risk of cardiovascular disease and cancer in later life: systematic review and meta-analysis,” BMJ (Clinical Research ed.), vol. 335, no. 7627, p. 974, 2007. [12] R. Orabona, E. Sciatti, E. Vizzardi et al., “Endothelial dys- function and vascular stiﬀness in women with previous pregnancy complicated by early or late pre-eclampsia,” Ul- trasound in Obstetrics & Gynecology, vol. 49, no. 1, pp. 116– 123, 2017. [13] B. Sibai, G. Dekker, and M. Kupferminc, “Pre-eclampsia,” Te Lancet, vol. 365, no. 9461, pp. 785–799, 2005. [14] A. Y. Bagrov and J. I. Shapiro, “Endogenous digitalis: path- ophysiologic roles and therapeutic applications,” Nature Clinical Practice Nephrology, vol. 4, no. 7, pp. 378–392, 2008. [15] S. W. Graves, R. Valdes Jr., B. A. Brown, A. B. Knight, and H. R. Craig, “Endogenous digoxin-immunoreactive substance in human pregnancies,” Te Journal of Clinical Endocrinology & Metabolism, vol. 58, no. 4, pp. 748–751, 1984. 5. Mineralocorticoid Antagonists Te fact that mineralocorticoid antagonists can oﬀset the eﬀects of digitoxin in rats has been ﬁrst reported by Selye [48, 49]. Subsequently, spironolactone and its active me- tabolite, canrenone, were reported to reverse digitalis tox- icity and to lower blood pressure in rat hypertension models, in which levels of CTS are elevated [50–52]. Most recently, spironolactone was reported to suppress cardiac ﬁbrosis in rats chronically treated by MBG [53]. Notably, in this study, MBG exhibited proﬁbrotic eﬀect in the absence of changes in aldosterone levels [53]. Importantly, high levels of MBG were associated with hypertension [33], stiﬀening of um- bilical vessels, and elevated vascular level collagen-1 in PE patients, and in vitro incubation of the healthy blood vessels Canrenone is the pharmacologically active metabolite of the spironolactone, used in hypertensive therapy. Canre- none exhibits its antialdosterone action because it blocks the binding of aldosterone to a cytosolic receptor in distal and collecting tubules of the nephron with subsequent inhibition of the synthesis of a speciﬁc protein that facilitates the entrance of Na+ ions into the cell and the consequent in- crement of NKA [61]. Several studies suggest that canrenone interacts with the ouabain-sensitive NKA competitively, by antagonizing the binding of 3H-ouabain [55] and MBG [53]. In addition to this well-known action, several studies have Cardiology Research and Practice 5 shown that canrenone was capable of inhibiting the NKA in vitro [50, 55, 62, 63]. Furthermore, if the pump was blocked by ouabain, canrenone could restimulate the pump [63, 64]. Interestingly, in experiment in rats, mineralocorticoid re- ceptor antagonists also could reverse MBG-induced eleva- tion of blood pressure associated with voluntary ethanol intake [65]. Acknowledgments Tis work was supported by the Russian Scientiﬁc Foun- dation grant no. 18-15-00222 (AYB, NIA, VAR). References It has been shown that canrenone is a partial competitor of ouabain in the human placental membranes. Moreover, the analysis of the Scatchard plot shows a unique intercept suggesting that canrenone and ouabain compete for the same site [52]. Te main ﬁndings of this study concern the eﬀect of canrenone on the binding of ouabain to placental NKA and the observation that canrenone, at therapeutic concentrations (about 8 to 10 μM), is able to reduce the ability of ouabain to inhibit the sodium pump [52]. Con- sidering the above data, it can be suggested that therapeutic eﬀects of canrenone can be a successful pathway in the treatment of PE due to its well-established antialdosterone action and to its modulatory eﬀects on the NKA activity. [1] National High Blood Pressure Education Program Working Group on High Blood Pressure in Pregnancy, “Report of the national high blood pressure education program working group on high blood pressure in pregnancy,” American Journal of Obstetrics & Gynecology, vol. 183, no. 1, pp. s1–s22, 2000. [1] National High Blood Pressure Education Program Working Group on High Blood Pressure in Pregnancy, “Report of the national high blood pressure education program working group on high blood pressure in pregnancy,” American Journal of Obstetrics & Gynecology, vol. 183, no. 1, pp. s1–s22, 2000. [2] ACOG, “Practice bulletin no. 125: chronic hypertension in pregnancy,” Obstetrics and Gynecology, vol. 119, no. 2, pp. 396–407, 2012. [3] S. Tangaratinam, A. Coomarasamy, F. O’Mahony et al., “Estimation of proteinuria as a predictor of complications of pre-eclampsia: a systematic review,” BMC Medicine, vol. 7, no. 1, p. 10, 2009. [4] F. G. 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https://openalex.org/W1993121297	OpenAlex	Open Science	CC-By	2,014	On the momentum dependence of the flavor structure of the nucleon sea	J. C. Peng	English	Spoken	6,511	12,576	See next page for additional authors Part of the Astrophysics and Astronomy Commons, and the Physics Commons Right click to open a feedback form in a new tab to let us know how this docum Right click to open a feedback form in a new tab to let us know how this docum On the Momentum Dependence of the Flavor Structure of the On the Momentum Dependence of the Flavor Structure of the Nucleon Sea Nucleon Sea Jen-Chieh Peng University of Illinois at Urbana-Champaign Wen-Chen Chang Academia Sinica, Taiwan University of Kentucky University of Kentucky UKnowledge UKnowledge Physics and Astronomy Faculty Publications Physics and Astronomy 9-7-2014 On the Momentum Dependence of the Flavor Structure of the On the Momentum Dependence of the Flavor Structure of the Nucleon Sea Nucleon Sea Jen-Chieh Peng University of Illinois at Urbana-Champaign Wen-Chen Chang Academia Sinica, Taiwan Hai-Yang Cheng Academia Sinica, Taiwan Tie-Jiun Hou Academia Sinica, Taiwan Keh-Fei Liu University of Kentucky, kehfei.liu@uky.edu See next page for additional authors Follow this and additional works at: https://uknowledge.uky.edu/physastron_facpub Part of the Astrophysics and Astronomy Commons, and the Physics Commons Right click to open a feedback form in a new tab to let us know how this document benefits you. Right click to open a feedback form in a new tab to let us know how this document benefits you. Repository Citation Repository Citation Peng, Jen-Chieh; Chang, Wen-Chen; Cheng, Hai-Yang; Hou, Tie-Jiun; Liu, Keh-Fei; and Qiu, Jian-Wei, "On the Momentum Dependence of the Flavor Structure of the Nucleon Sea" (2014). Physics and Astronomy Faculty Publications. 351. https://uknowledge.uky.edu/physastron_facpub/351 This Article is brought to you for free and open access by the Physics and Astronomy at UKnowledge. It has been accepted for inclusion in Physics and Astronomy Faculty Publications by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. University of Kentucky University of Kentucky UKnowledge UKnowledge University of Kentucky University of Kentucky UKnowledge UKnowledge Physics and Astronomy Faculty Publications Physics and Astronomy Faculty Publications Physics and Astronomy Faculty Publications Physics and Astronomy 9-7-2014 On the Momentum Depende On the Momentum Depende Nucleon Sea Nucleon Sea Jen-Chieh Peng University of Illinois at Urbana-Champaign Wen-Chen Chang Academia Sinica, Taiwan Hai-Yang Cheng Academia Sinica, Taiwan Tie-Jiun Hou Academia Sinica, Taiwan Keh-Fei Liu University of Kentucky, kehfei.liu@uky.edu See next page for additional authors Follow this and additional works at: https://u Part of the Astrophysics and Astronomy Right click to open a feedback form in a Right click to open a feedback form in a Repository Citation Repository Citation Peng, Jen-Chieh; Chang, Wen-Chen; Cheng, H Momentum Dependence of the Flavor Struct Faculty Publications. 351. https://uknowledge.uky.edu/physastron_fac This Article is brought to you for free and open ac accepted for inclusion in Physics and Astronomy UKnowledge. For more information, please conta Notes/Citation Information Notes/Citation Information Published in Physics Letters B, v. 736, p. 411-414. © 2014 The Authors. Published by Elsevier B.V. © 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). Funded by SCOAP3. Repository Citation Repository Citation Peng, Jen-Chieh; Chang, Wen-Chen; Cheng, Hai-Yang; Hou, Tie-Jiun; Liu, Keh-Fei; and Qiu, Jian-Wei, "On the Momentum Dependence of the Flavor Structure of the Nucleon Sea" (2014). Physics and Astronomy Faculty Publications. 351. h k l d k d h f b 3 https://uknowledge.uky.edu/physastron_facpub/351 This Article is brought to you for free and open access by the Physics and Astronomy at UKnowledge. It has been accepted for inclusion in Physics and Astronomy Faculty Publications by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. On the Momentum Dependence of the Flavor Structure of the Nucleon Sea On the Momentum Dependence of the Flavor Structure of the Nucleon Sea Digital Object Identifier (DOI) http://dx.doi.org/10.1016/j.physletb.2014.07.050 Digital Object Identifier (DOI) http://dx.doi.org/10.1016/j.physletb.2014.07.050 a r t i c l e i n f o Article history: Received 16 May 2014 Received in revised form 1 July 2014 Accepted 28 July 2014 Available online 1 August 2014 Editor: D.F. Geesaman Keywords: Parton distributions Sea quark ¯d(x) −¯u(x) Lattice QCD Difference between the ¯u and ¯d sea quark distributions in the proton was ﬁrst observed in the violation of the Gottfried sum rule in deep-inelastic scattering (DIS) experiments. The parton momentum fraction x dependence of this difference has been measured over the region 0.02 < x < 0.35 from Drell–Yan and semi-inclusive DIS experiments. The Drell–Yan data suggested a possible sign-change for ¯d(x) −¯u(x) near x ∼0.3, which has not yet been explained by existing theoretical models. We present an independent evidence for the ¯d(x) −¯u(x) sign-change at x ∼0.3 from an analysis of the DIS data. We further discuss the x-dependence of ¯d −¯u in the context of meson cloud model and the lattice QCD formulation. © 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). Funded by SCOAP3. Article history: Received 16 May 2014 Received in revised form 1 July 2014 Accepted 28 July 2014 Available online 1 August 2014 Editor: D.F. Geesaman Keywords: Parton distributions Sea quark ¯d(x) −¯u(x) Lattice QCD © 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). Funded by SCOAP3. It is now a well established fact that the ¯u and ¯d distribu- tions in the proton are strikingly different. The ﬁrst evidence for this difference came from the observation of the violation of the Gottfried sum rule [1] in a deep-inelastic scattering (DIS) ex- periment by the NMC Collaboration [2]. The Gottfried sum rule, IG ≡ 1 0 [F p 2 (xB) −F n 2(xB)]/xB dxB = 1/3, is obtained under the as- sumption of a symmetric ¯u and ¯d sea [1], where xB is the Bjorken variable and is effectively equal to parton momentum fraction x probed in DIS using the leading order QCD factorization formal- ism of the structure function F2(xB). The NMC measurement of IG = 0.235 ± 0.026 implies that this assumption is invalid with an x-integrated difference of 1 0 [¯d(x) −¯u(x)]dx = 0.148 ± 0.039. distribution functions (PDFs). Fig. 1 compares the data measured at Q 2 = 54 GeV2 from Fermilab E866 with parameterizations of several PDFs. Jen-Chieh Peng a, Wen-Chen Chang b, Hai-Yang Cheng b, Tie-Jiun Hou b, Keh-Fei Liu c, Jian-Wei Qiu d,e a Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA b Institute of Physics, Academia Sinica, Taipei 11529, Taiwan c Department of Physics and Astronomy, University of Kentucky, Lexington, KY 40506, USA d Physics Department, Brookhaven National Laboratory, Upton, NY 11973, USA e C.N. Yang Institute for Theoretical Physics and Department of Physics and Astronomy, Stony Brook University, Stony Brook, NY 11794, USA a Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA b Institute of Physics, Academia Sinica, Taipei 11529, Taiwan c Department of Physics and Astronomy, University of Kentucky, Lexington, KY 40506, USA d Physics Department, Brookhaven National Laboratory, Upton, NY 11973, USA e C.N. Yang Institute for Theoretical Physics and Department of Physics and Astronomy, Stony Broo a Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA b Institute of Physics, Academia Sinica, Taipei 11529, Taiwan f y , , p , c Department of Physics and Astronomy, University of Kentucky, Lexington, KY 40506, USA d Physics Department, Brookhaven National Laboratory, Upton, NY 11973, USA y p , y, p , , e C.N. Yang Institute for Theoretical Physics and Department of Physics and Astronomy, Stony Brook University, Stony Brook, NY 11794, USA http://dx.doi.org/10.1016/j.physletb.2014.07.050 0370-2693/© 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). Funded by SCOAP3. Authors Authors Jen-Chieh Peng, Wen-Chen Chang, Hai-Yang Cheng, Tie-Jiun Hou, Keh-Fei Liu, and Jian-Wei Qiu This article is available at UKnowledge: https://uknowledge.uky.edu/physastron_facpub/351 This article is available at UKnowledge: https://uknowledge.uky.edu/physastron_facpub/35 Physics Letters B 736 (2014) 411–414 Contents lists available at ScienceDirect of the nucleon sea Jen-Chieh Peng a, Wen-Chen Chang b, Hai-Yang Cheng b, Tie-Jiun Hou b, Keh-Fei Liu c, Jian Wei Qiu d,e Jen-Chieh Peng a, Wen-Chen Chang b, Hai-Yang Cheng b, Tie-Jiun Hou b, Keh-Fei Jian-Wei Qiu d,e Jen-Chieh Peng a, Wen-Chen Chang b, Hai-Yang Cheng b, Tie-Jiun Hou b, Keh-Fei Liu c Jian-Wei Qiu d,e shed by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). Funded by http://dx.doi.org/10.1016/j.physletb.2014.07.050 0370-2693/© 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativeco SCOAP3. a r t i c l e i n f o Future measurements of ¯d(x)/¯u(x) at x > 0.25 in Drell–Yan experiments could provide strong constraints and new insights on the origins of the ﬂavor structure of the proton’s sea. g g p The NMC measurement of the Gottfried sum involves the F2 structure functions on proton and neutron. In terms of QCD factor- ization, we have at the leading order in αs, Although Fig. 2 shows similar trends for the x-dependence of ¯d −¯u extracted from the E866 Drell–Yan and the NMC DIS data, these two data sets correspond to two different Q 2 scales. A more direct comparison can be obtained by analyzing the ﬁnal results published by the NMC Collaboration on the ratio R(x) = F d 2(x)/F p 2 (x) [20]. The values of F p 2 (x) −F n 2(x) could be calculated from 2F d 2(x) ∗(1/R(x) −1/rd N(x)), by using the parametrization of F d 2(x) of Ref. [21]. The rd N(x) is the ratio of deuteron to isoscalar nucleon structure functions F d 2(x) = rd N(x) ∗(F p 2 (x) + F n 2(x))/2 and we use rd N(x) of the CJ12mid set at Q 2 = 100 GeV2 [22] for the evaluation. Refs. [20] and [21] included not only additional data that were not available for NMC’s earlier evaluation of the Got- tfried sum [2], but also the values of R(x) at different bins of Q 2 ranging from 0.16 to 99.03 GeV2. The high Q 2 data makes it possible to compare the E866 Drell–Yan data on ¯d(x) −¯u(x) at Q 2 = 54 GeV2 with that evaluated using Eq. (2) and NMC data at a similar Q 2. As the E866 Drell–Yan data on ¯d(x) −¯u(x) correspond to Q 2 = 54 GeV2, a comparison could be made by using the NMC data at similar Q 2. The mean values of Q 2 for the four highest Q 2 bins of NMC data are around 34, 45, 63, and 95 GeV2. Fig. 3 shows ¯d(x) −¯u(x) for these four values of Q 2 using Eq. 2 with the JR14 parametrization of the valence quark distributions and the NMC data [20] for F p 2 (x) −F n 2(x). The uncertainties of both R(x) and the parametrization of F d 2 have been included in the evaluation of F p 2 (x) −F n 2(x). Fig. a r t i c l e i n f o We have also performed calculations with other sets of recent PDFs, obtained very similar results and reached the same conclusion. In Fig. 2, we show, for example, the values of ¯d(x) −¯u(x) (ﬁlled stars) obtained by using uv(x) −dv(x) of the CT10 PDF parametrization [7] along with the same NMC data. The values of ¯d(x) −¯u(x) obtained by using CT10 and JR14 are practi- cally identical for x > 0.2. This ﬁnding is effectively a consequence of the fact that the uv(x) −dv(x) distribution in Eq. (2) is well constrained by QCD global ﬁt of the extensive DIS and hadronic scattering data. of ¯d(x) −¯u(x) at Q 2 = 54 GeV2 (ﬁlled squares) derived by the E866 Collaboration [4]. The sign-change of ¯d −¯u at x ∼0.3 as indicated by the E866 data is clearly consistent with the behav- ior of open circles obtained by using Eq. (2) based on the NMC data and the JR14 PDFs. Although the JR14 uses a parametrization of ¯d −¯u that is positive at all x, as shown in Fig. 1, we demon- strated in Fig. 2 that NMC data together with the valence quark distributions of JR14 could lead to a sign-change of ¯d(x) −¯u(x) distribution at x ∼0.3. We have also performed calculations with other sets of recent PDFs, obtained very similar results and reached the same conclusion. In Fig. 2, we show, for example, the values of ¯d(x) −¯u(x) (ﬁlled stars) obtained by using uv(x) −dv(x) of the CT10 PDF parametrization [7] along with the same NMC data. The values of ¯d(x) −¯u(x) obtained by using CT10 and JR14 are practi- cally identical for x > 0.2. This ﬁnding is effectively a consequence of the fact that the uv(x) −dv(x) distribution in Eq. (2) is well constrained by QCD global ﬁt of the extensive DIS and hadronic scattering data. In this paper we address the intriguing possibility that d −¯u changes sign at the x ∼0.3 region. We ﬁrst show that an indepen- dent experimental evidence for this sign-change, other than the one shown in Fig. 1 from the Drell–Yan data, comes from an anal- ysis of the NMC DIS data. We then discuss the signiﬁcance of this sign-change and the stringent constraint it imposes on theoreti- cal models. We also discuss the implications on the x-dependence of ¯d −¯u using the lattice QCD formulation for the sea-quark par- ton distributions. a r t i c l e i n f o The open circles and ﬁlled stars correspond to results obtained with the JR14 and CT10 PDFs, respectively. Also shown are the values of ¯d(x) −¯u(x) at Q 2 = 54 GeV2 from the Fermilab E866 experiment. Fig. 1. Ratio of ¯d(x) over ¯u(x) versus Bjorken-x from experiments NA51 [3] and E866 [4]. Parameterizations from several parton distribution functions are also shown. Fig. 1. Ratio of ¯d(x) over ¯u(x) versus Bjorken-x from experiments NA51 [3] and E866 [4]. Parameterizations from several parton distribution functions are also shown. Fig. 2. Values of ¯d(x) −¯u(x) at Q 2 = 4 GeV2 evaluated using Eq. (2), as discussed in the text. The open circles and ﬁlled stars correspond to results obtained with the JR14 and CT10 PDFs, respectively. Also shown are the values of ¯d(x) −¯u(x) at Q 2 = 54 GeV2 from the Fermilab E866 experiment. Fig. 2. Values of ¯d(x) −¯u(x) at Q 2 = 4 GeV2 evaluated using Eq. (2), as discussed in the text. The open circles and ﬁlled stars correspond to results obtained with the JR14 and CT10 PDFs, respectively. Also shown are the values of ¯d(x) −¯u(x) at Q 2 = 54 GeV2 from the Fermilab E866 experiment. Fig. 1. Ratio of ¯d(x) over ¯u(x) versus Bjorken-x from experiments NA51 [3] and E866 [4]. Parameterizations from several parton distribution functions are also shown. accuracy and to extend the kinematic coverage of the ¯d/¯u mea- surement to the x > 0.3 region. This is the goal of an ongoing Fermilab Drell–Yan experiment, E906 [16], and a proposed experi- ment [17] at the J-PARC facility. The x-dependence of ¯d/¯u (or the related quantity ¯d −¯u) at large x remains a topics of much interest both theoretically and experimentally. of ¯d(x) −¯u(x) at Q 2 = 54 GeV2 (ﬁlled squares) derived by the E866 Collaboration [4]. The sign-change of ¯d −¯u at x ∼0.3 as indicated by the E866 data is clearly consistent with the behav- ior of open circles obtained by using Eq. (2) based on the NMC data and the JR14 PDFs. Although the JR14 uses a parametrization of ¯d −¯u that is positive at all x, as shown in Fig. 1, we demon- strated in Fig. 2 that NMC data together with the valence quark distributions of JR14 could lead to a sign-change of ¯d(x) −¯u(x) distribution at x ∼0.3. a r t i c l e i n f o The E866 data show the salient feature that ¯d/¯u rises linearly with x for x < 0.15 and then drops as x further in- creases. At the largest value of x (x = 0.315), the ¯d/¯u ratio falls below unity, albeit with large experimental uncertainty. This in- triguing x-dependence of ¯d/¯u is reﬂected in recent PDFs including CTEQ6 [6], CT10 [7], MSTW08 [8], and JR14 [9]. However, for the CTEQ4M [10] PDF, which predated the E866 data, the ¯d/¯u ra- tios at large x are not well described by the parameterizations. In particular, ¯d(x)/¯u(x) remains greater than unity, or equivalently, ¯d(x) −¯u(x) > 0, at all x. The parameterizations of the more re- cent PDFs are suﬃciently ﬂexible to accommodate a sign-change for ¯d(x) −¯u(x) at x ∼0.3, as suggested by the E866 data. 0 The NMC result was subsequently checked using two indepen- dent experimental techniques. From measurements of the Drell– Yan cross section ratios of [σ(p +d)]/[σ(p + p)], the NA51 [3] and the Fermilab E866 [4] experiments measured ¯d/¯u as a function of x over the kinematic range of 0.015 < x < 0.35. As shown in Fig. 1, the ¯d/¯u ratios clearly differ from unity. From a semi-inclusive DIS measurement, the HERMES Collaboration also reported the obser- vation [5] of ¯d(x) −¯u(x) ̸= 0, consistent with the Drell–Yan results. The ¯d(x)/¯u(x) data obtained from the Drell Yan experiments Many theoretical models have been put forward to explain the surprisingly large difference between ¯d(x) and ¯u(x). For reviews on various theoretical models, see Refs. [11–15]. While these models can explain the enhancement of ¯d over ¯u involving various mecha- nisms such as meson cloud, chiral-quark, intrinsic sea, soliton, and Pauli-blocking, none of them predicts that the ¯d/¯u ratio falls be- low unity at any value of x [14]. In order to understand the origin of the sea-quark ﬂavor structure, it is important to improve the The d(x)/¯u(x) data obtained from the Drell–Yan experiments have provided stringent constraints for parameterizing the parton J.-C. Peng et al. / Physics Letters B 736 (2014) 411–414 412 Fig. 1. Ratio of ¯d(x) over ¯u(x) versus Bjorken-x from experiments NA51 [3] and E866 [4]. Parameterizations from several parton distribution functions are also shown. Fig. 2. Values of ¯d(x) −¯u(x) at Q 2 = 4 GeV2 evaluated using Eq. (2), as discussed in the text. a r t i c l e i n f o 3 shows that the values of ¯d(x) −¯u(x) at x > 0.3 are mostly negative with the mean values of −0.009 ± 0.006, −0.012 ± 0.006, −0.016 ± 0.008, and −0.001 ± 0.008, respectively, F p 2 (x) −F n 2(x) = 1 3 x u(x) + ¯u(x) −d(x) −¯d(x) , (1) (1) where x = xB was used at this order. Eq. (1) is obtained under the usual assumption of charge symmetry of parton distributions and the equality of heavy-quark (s, c, b) distributions in proton and neutron. Note that the Q 2 dependence in F p,n 2 (x, Q 2) and parton distributions q(x, Q 2) is implicit. The magnitude of order α1 s and α2 s perturbative QCD effect is estimated to be small, on the order of 0.2% at Q = 10 GeV [18]. From Eq. (1) and the deﬁnition of valence quarks, uv(x) = u(x) −¯u(x) and dv(x) = d(x) −¯d(x), one readily obtains the following expression: ¯d(x) −¯u(x) = 1 2 uv(x) −dv(x) −3 2x F p 2 (x) −F n 2(x) . (2) (2) Eq. (2) shows that the x dependence of ¯d −¯u can be extracted from the NMC measurement of F p 2 (x) −F n 2(x) and the parametriza- tion of uv(x) −dv(x) from various PDFs. To illustrate this, we show in Fig. 2 the values of ¯d(x) −¯u(x) at Q 2 = 4 GeV2 using Eq. (2), where the ﬁrst term of the right-hand side, uv(x) −dv(x), is taken from the NNLO JR14 parametrization [9] and the second term, F p 2 (x) −F n 2(x), is taken from the NMC data [2] at Q 2 = 4 GeV2. The JR14 is a recent PDF where the nuclear corrections from the CJ group [19] is implemented and ¯d(x) −¯u(x) > 0 is assumed at all x in the global analysis. We also show in Fig. 2 the values Eq. (2) shows that the x dependence of ¯d −¯u can be extracted from the NMC measurement of F p 2 (x) −F n 2(x) and the parametriza- tion of uv(x) −dv(x) from various PDFs. To illustrate this, we show in Fig. 2 the values of ¯d(x) −¯u(x) at Q 2 = 4 GeV2 using Eq. a r t i c l e i n f o (2), where the ﬁrst term of the right-hand side, uv(x) −dv(x), is taken from the NNLO JR14 parametrization [9] and the second term, F p 2 (x) −F n 2(x), is taken from the NMC data [2] at Q 2 = 4 GeV2. The JR14 is a recent PDF where the nuclear corrections from the CJ group [19] is implemented and ¯d(x) −¯u(x) > 0 is assumed at all x in the global analysis. We also show in Fig. 2 the values J.-C. Peng et al. / Physics Letters B 736 (2014) 411–414 413 Fig. 3. Values of ¯d(x) −¯u(x) evaluated using Eq. (2) and the NMC data [20,21] of R(x) and F d 2(x) at the four largest values of Q 2. The JR14 parametrization for uv(x) −dv(x) at the corresponding Q 2 and the ratio of deuteron to isoscalar nu- cleon structure functions rd N(x) of the CJ12mid set at Q 2 = 100 GeV2 [22] are used. The values of ¯d(x) −¯u(x) from E866 measurement at Q 2 = 54 GeV2 are also shown. The solid curves are ¯d(x) −¯u(x) from JR14. Fig. 4. Two gauge invariant and topologically distinct diagrams for (a) connected sea (left graph) and (b) disconnected sea (right graph). Fig. 4. Two gauge invariant and topologically distinct diagrams for (a) connected sea (left graph) and (b) disconnected sea (right graph). sign at large x. The same conclusion can be obtained for the chi- ral quark model [27–29], in which the pions couple directly to the constituent quarks. Since there are two u quarks coupling to π+ (u →π+ + d) and only one d quark coupling to π−(d →π−+ u), the larger probability for the π+ meson cloud relative to the π− cloud would lead to ¯d > ¯u for all x. Similar conclusions can be obtained in the intrinsic sea model [30], the chiral-quark soliton model [31], and the statistical model [32]. ¯ To shed some light on the x-dependence of d −¯u, we consider the origins of sea quarks in the lattice QCD approach. There are two sources for the ¯d and ¯u seas in the path-integral formalism of the hadronic tensor deﬁning the structure function F2(x) [33], as shown in the two gauge-invariant and topologically distinct di- agrams in Fig. 4. One is the connected sea (CS) from the connected insertion diagram (Fig. a r t i c l e i n f o 4(a)) and the other is the disconnected sea (DS) from the disconnected insertion diagram (Fig. 4(b)). For the case with isospin symmetry, i.e. mu = md, it is shown [34] that the DS does not distinguish ¯u from ¯d. Hence, the ¯u(x), ¯d(x) dif- ference must originate solely from the CS (Fig. 4(a)). It is well known that sea quark distribution generated by the disconnected diagram is a steeply falling function of momentum fraction x, be- cause the gluon radiated from the initial quark line is dominantly soft due to the ∝1/x behavior of the splitting kernel. In contrast, sea quarks generated by the connected diagram have an x−1/2 be- havior at small x and are most relevant in the medium and large x region. While lattice QCD so far could only generate the moments rather than the x-dependence of quark distributions, a ﬁrst attempt to separate the CS and DS components of the ¯u(x) + ¯d(x) was re- ported recently [35]. The extracted CS and DS for ¯u(x) + ¯d(x) [35] have a distinct x dependence in qualitative agreement with expec- tation. The DS dominates the small x region (i.e. x < 0.05) while the CS dominates the x > 0.05 region. Fig. 3. Values of ¯d(x) −¯u(x) evaluated using Eq. (2) and the NMC data [20,21] of R(x) and F d 2(x) at the four largest values of Q 2. The JR14 parametrization for uv(x) −dv(x) at the corresponding Q 2 and the ratio of deuteron to isoscalar nu- cleon structure functions rd N(x) of the CJ12mid set at Q 2 = 100 GeV2 [22] are used. The values of ¯d(x) −¯u(x) from E866 measurement at Q 2 = 54 GeV2 are also shown. The solid curves are ¯d(x) −¯u(x) from JR14. for the four Q 2 bins. The agreement between the E866 and NMC results is now improved when compared with Fig. 2. In particu- lar, both the NMC and the E866 experiments show evidence that ¯d(x) −¯u(x) changes sign at x ∼0.3. Since both the NMC data and the E866/NA51 Drell–Yan data are included in recent global ﬁts for determining the parton dis- tributions, it is conceivable that the NMC data have already played a role in constraining the behavior of ¯d(x) −¯u(x) at large x. a r t i c l e i n f o Nev- ertheless, the possible sign-change of ¯d(x) −¯u(x) for x ∼0.3 has only been attributed in the literature to the E866 data, which have large uncertainty at the highest x region. We show that an inde- pendent indication for this sign-change is already provided by the NMC DIS data, which were obtained prior to the E866 Drell–Yan data. The signiﬁcance of the sign-change of ¯d(x) −¯u(x) for x > 0.3, if conﬁrmed by future experiments, is that it would severely chal- lenge existing theoretical models which can successfully explain ¯d(x) −¯u(x) at x < 0.25, but predict no sign-change at higher x. Take for example the meson-cloud model [23–26], which treats proton as a linear combination of a bare proton plus pion–nucleon and pion–delta Fock states: It is instructive to consider ¯d(x) −¯u(x) in three different x regions. At small x, the DS with small x behavior of x−1 dom- inates. For Q 2 = 2.5 GeV2 where the CS and DS are explicitly separated [35], this is the region where x < 0.05. Since the only difference between ¯uDS and ¯dDS is the u/d mass difference which is much smaller than the scale for the validity of the parton picture, we expect the difference between them due to isospin symmetry breaking to be very small. In the mid-x region (from x = 0.05 to x ∼1/3), dominated by the CS with an x-dependence of x−1/2, the Fock space wavefunction of the quarks in the nucleon is important. It is in this region that the DIS and Drell–Yan experiments reveal that ¯u(x) < ¯d(x). The dominance of the CS at this region of x sug- gests a greater chance for the CS partons to share the momentum with the valence quarks resulting in the meson–baryon conﬁgura- tions. Hence the pronounced feature of ¯u(x) < ¯d(x) in this x region can be understood in terms of the pion cloud model. \|p⟩→ 1 −a2 −b2 \|p0⟩ + a − 1 3 p0π 0 + 2 3 n0π+ + b 1 2 ++ 0 π− − 1 3 + 0 π 0 + 1 6 0 0π+ . (3) (3) The subscript zeros denote bare baryons with ﬂavor symmetric seas. The ¯u and ¯d seas have contributions from the valence an- tiquarks of the pion cloud, i.e., ¯d in π+ and ¯u in π−. a r t i c l e i n f o The pion– nucleon amplitude is larger than the pion–delta amplitude (a > b) due to the heavier mass for the . The excess of ¯d over ¯u arises because of the dominance of the n0π+ conﬁguration over the less probable ++ 0 π−conﬁguration. This leads to an overall excess of ¯d over ¯u. Moreover, the x distribution for ¯u is softer than that of ¯d, since π−in the ++ 0 π−conﬁguration carries a smaller fraction of the proton’s momentum than π+ in the n0π+ conﬁguration. As a consequence, ¯d(x) −¯u(x) remains positive and does not change At even larger x (x > 1/3) the nature of the sea quarks is ex- pected to be strongly inﬂuenced by the valence quarks. Intuitively, the connected sea diagram provides a natural mechanism for gen- erating more ¯u(x) than ¯d(x) at this region, since there are two u valence quarks capable of generating ¯u quarks as shown in Fig. 4. Fig. 5 shows speciﬁc examples of diagrams responsible for gener- ating antiquarks from one (top) or two (bottom) valence quarks. J.-C. Peng et al. / Physics Letters B 736 (2014) 411–414 414 Fig. 5. QCD quantum ﬂuctuation capable of generating connected ¯u(x) or ¯d(x), in- volving one (top) or two (bottom) valence quarks, which could lead to more ¯u(x) than ¯d(x). ture function of the hadronic tensor or the recently proposed direct calculation via a Lorentz boost [38,39]. When reliable results are obtained, they would provide a direct check on the possible sign- change for ¯d(x) −¯u(x) at x ∼0.3. New experimental information on the x-dependence of the ¯d −¯u at large x, anticipated for future Drell–Yan experiments, together with comprehensive global anal- yses would be critical for understanding the origins of the ﬂavor structure of the nucleon sea. Acknowledgements The quantum ﬂuctuation could be thought as a time sequence of four steps: 1) ﬂuctuation of a valence quark into a quark and a highly vir- tual gluon, 2) a quick splitting of the gluon into a quark and antiquark pair, 3) annihilation or recombination of the quark and the newly produced antiquark into a highly virtual gluon, which is then, 4) absorbed by the quark. Since both valence u and d quarks can go through the same QCD quantum ﬂuctuation to generate ¯u and ¯d quarks, the mechanism depicted in Fig. 5 could generate about twice of ¯u(x) over ¯d(x) due to the 2-to-1 ratio of valence quarks. But, this ﬂuctuation is the most probable only if partons involved have an excellent coalescence, and therefore, it should be very short-lived. That is, it is unlikely to generate enough imbal- ance between ¯u and ¯d to compete with what could be generated by the pion cloud or other mechanisms/models at small-x. How- ever, this mechanism is not very sensitive to the parent quark’s momentum fraction x, and would become relevant when the im- balance generated by other mechanisms/models dies away at large x. It is noted that ¯u > ¯d was also suggested by a model calculation examining the antisymmetrization effect of the nucleon sea aris- ing from gluon exchange between conﬁned valance quarks [36]. The data indicates that such transition takes place at x ∼1/3. A detailed calculation of ¯u(x) and ¯d(x) in terms of connected (or recombination) diagrams, like that in Ref. [37], is beyond the scope of this letter, and will be presented later. Acknowledgements We acknowledge helpful discussion with Jiunn-Wei Chen and Chien-Peng Yuan. This work was supported in part by the Na- tional Science Council of the Republic of China and the U.S. De- partment of Energy under contact Nos. DE-AC02-98CH10886 and DE-FG05-84ER40154, and the National Science Foundation under grant Nos. PHY-0969739 and PHY-1316617. Fig. 5. QCD quantum ﬂuctuation capable of generating connected ¯u(x) or ¯d(x), in- volving one (top) or two (bottom) valence quarks, which could lead to more ¯u(x) than ¯d(x). In Fig. 5, the antiquark mode of the QCD quantum ﬂuctuation of a quark is probed by the currents Jμ and Jν. The quantum ﬂuctuation could be thought as a time sequence of four steps: 1) ﬂuctuation of a valence quark into a quark and a highly vir- tual gluon, 2) a quick splitting of the gluon into a quark and antiquark pair, 3) annihilation or recombination of the quark and the newly produced antiquark into a highly virtual gluon, which is then, 4) absorbed by the quark. Since both valence u and d quarks can go through the same QCD quantum ﬂuctuation to generate ¯u and ¯d quarks, the mechanism depicted in Fig. 5 could generate about twice of ¯u(x) over ¯d(x) due to the 2-to-1 ratio of valence quarks. But, this ﬂuctuation is the most probable only if partons involved have an excellent coalescence, and therefore, it should be very short-lived. That is, it is unlikely to generate enough imbal- ance between ¯u and ¯d to compete with what could be generated by the pion cloud or other mechanisms/models at small-x. How- ever, this mechanism is not very sensitive to the parent quark’s momentum fraction x, and would become relevant when the im- balance generated by other mechanisms/models dies away at large x. It is noted that ¯u > ¯d was also suggested by a model calculation examining the antisymmetrization effect of the nucleon sea aris- ing from gluon exchange between conﬁned valance quarks [36]. The data indicates that such transition takes place at x ∼1/3. A detailed calculation of ¯u(x) and ¯d(x) in terms of connected (or recombination) diagrams, like that in Ref. [37], is beyond the scope of this letter, and will be presented later. 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In summary, we have discussed the importance of the possi- ble sign-change for ¯d(x) −¯u(x) at x ∼0.3 for understanding the ﬂavor structure of the nucleon sea. We present an independent evidence for the ¯d(x) −¯u(x) sign-change at large x from an anal- ysis of existing DIS data. This sign-change cannot be explained by any existing theoretical model on the nucleon sea. Nevertheless, a qualitative explanation for the sign-change at large x is provided in the context of lattice QCD formalism. Up to now, only the con- nection between the NMC data and the integral of ¯d(x) −¯u(x) has been discussed. The current work hopefully would lead to some dedicated studies by the various PDF groups to assess the impact of the NMC data on the x-dependence of ¯d −¯u. We note that ¯d(x) −¯u(x) can be calculated on the lattice [33] from the struc- [26] W.P. Hwang, J. Speth, G.E. Brown, Z. Phys. A 339 (1991) 38 [26] W.P. Hwang, J. Speth, G.E. Brown, Z. Phys. A 339 (1991) 383. [27] E. Eichten, I. 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https://openalex.org/W2410272654	OpenAlex	Open Science	CC-By	2,016	Efficient cardinality estimation for k-mers in large DNA sequencing data sets	Luiz Irber	English	Spoken	3,013	5,445	F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint Luiz C. Irber Jr.1 and C. Titus Brown2 Luiz C. Irber Jr.1 and C. Titus Brown2 1lcirberjr@ucdavis.edu, Department of Population Health and Reproduction, University of California, Davis, Davis, CA 95616, USA 2 2ctbrown@ucdavis.edu, corresponding author, Department of Population Health and Reproduction, University of California, Davis, Davis, CA 95616, USA Introduction DNA sequencing technologies have increased in data gen- eration capacity faster than Moore’s Law for more than a decade now, driving the development of new computa- tional analysis approaches. Add(x, M) : M[n] ←ρ(f (x) ≫p) where ρ(h) is the number of leading zeros in the binary representation of h. Alon et al. [1996] analyses randomized algorithms for the approximation of “frequency moments” of a sequence us- ing a streaming approach, where items of the sequence are not (or can not be) stored, and are processed se- quentially (arrive one by one). KmerStream Melsted and Halldórsson [2014] implemented The cardinality estimator E is the normalized harmonic mean of the estimation on the substream: E = αmm2 Pm j=1 2−M[j] , with αm = m Z ∞ 0 log2 2 + u 1 + u m du − Multiple HLL sketches can be merged by taking the max- imum value, element-wise, from every sketch byte-array. For a more detailed description and error bounds analysis, see [Flajolet et al., 2008]. A number of probabilistic data structures and algorithms have been developed over the last few years to scale anal- ysis approaches to the rapidly increasing volume of data [Pell et al., 2012], Here we present an open implementation of the Hyper- LogLog cardinality estimation algorithm, specialized for k-length substrings of DNA strings, or k-mers. The Hyper- LogLog sketch (HLL) [Flajolet et al., 2008] is a cardinality estimation data structure with constant (and low) mem- ory footprint. Methods We implemented HyperLogLog for k-mers in C++ on top of the khmer library. khmer[Crusoe et al.] is a library and suite of command line tools for working with DNA se- quences. It implements k-mer counting, read ﬁltering and graph traversal using probabilistic data structures such as Bloom Filters and Count-Min Sketches. Building on top of khmer leveraged the existing infrastructure (read parsers, package installation, API signatures and some k-mer hash- ing methods). Our implementation of HLL for multiple processors uses a shared memory model, creating multiple HLL sketches in order to avoid synchronization and locking when adding elements. A master thread processes the input and dis- tributes reads between task threads. After all the reads are consumed the sketches are merged and the ﬁnal sketch can be used for cardinality estimation of the entire data set. Since sketch sizes are small (16 KiB for a 1% error rate), instantiating additional temporary HLL sketches is a viable tradeoff. One alternative is one HLL shared be- tween threads, with a locking mechanism to isolate the byte array on updates. This would avoid the merge pro- cess at the end, but then threads could block on updating the shared structure. The HyperLogLog sketch (HLL) [Flajolet et al., 2008] es- timates the approximate cardinality (F0 frequency mo- ment) of a set. The HLL is composed of a byte array M[1..m] initialized with 0s and a precision value p, where m = 2p The expected error rate e is e = 1.04 pm m = 2p The expected error rate e is e = 1.04 pm The shared memory model is also a good ﬁt since this is the architecture most potential users have readily avail- able for use. OpenMP is an appropriate choice for the conditions we outlined, and the code compiles to a se- quential implementation when OpenMP is not available. We used the OpenMP tasks functionality, since they map well to our problem. and by modifying p we can control the precision of the estimate. Each position of M represents the longest run of zeros found in the nth substream, where n is an index calculated from the least signiﬁcant p bits of the hashed value: Implementation details We chose MurmurHash3 for the hash function because it is one of the fastest non-cryptographic hash functions available and it has a reasonably uniform hash space dis- tribution. Since a k-mer is a representation of a substring of a single strand of DNA, the reverse complement on the other strand must also be considered to avoid overcount- ing. We hash the k-mer and its reverse complement indi- vidually using MurmurHash3 and create a unique value by doing a binary exclusive-OR on the two hashed values, generating a 128-bit hash value. For compatibility with the current khmer codebase, where hashed values are 64 bits long, we do another binary exclusive-OR over the ﬁrst and last 64 bits to have a single 64-bit hash value. This procedure is executed len(sequence) −(k −1) times for each sequence in the dataset, where k is the desired k- mer size. Efﬁcient k-mer cardinality estimation is useful for a vari- ety of purposes, including estimating the memory require- ments for de Bruijn graph assemblers [Zerbino and Bir- ney, 2008] and choosing the initial memory allocation for data structures like Bloom ﬁlters and Count-Min Sketches ([Zhang et al., 2014]). Abstract We present an open implementation of the HyperLogLog cardinality estimation sketch for counting ﬁxed-length substrings of DNA strings (“k-mers”). g g g g ( ) The HyperLogLog sketch implementation is in C++ with a Python interface, and is distributed as part of the khmer software package. khmer is freely available from https://github.com/dib-lab/khmer under a BSD License. The features presented here are included in version 1.4 and later. Page 1 of 5 Page 1 of 5 . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) hash value using a hash function f , hash value using a hash function f , Grant information This work was supported by grant 2013-67015-21357 from the United States Department of Agriculture. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The error in the HLL implementation is close to the ideal upper bound, 1%. The difference can be attributed to MurmurHash3 and our hashing procedure not being per- fectly uniform on the hash value space. Author contributions LCI and CTB conceived the study and designed the ex- periments. LCI carried out the research, prepared ﬁgures and tables and performed the computation work. LCI and CTB analyzed the data and wrote the paper. All authors were involved in the revision of the draft manuscript and have agreed to the ﬁnal content. • SRR1216679, containing 675,236 Illumina reads with average length of 250 bp, 168,809,000 base- pairs and 17,510,301 unique k-mers. Referred to be- low as the medium dataset. Tables 1 and 2 show the runtime, memory consumption, cardinality and the error of the estimate compared to the true cardinality of each dataset. Both exact implemen- tations report the same cardinality and consume similar amounts of memory for each dataset, with the Python implementation taking longer to run. HLL is an order of magnitude faster and consumes a constant amount of memory for both cases, about 6 times less on the small dataset and 200 times less on the medium dataset. Streaming RAM and a SATA block storage with 200 MB/s transfer rate. During the streaming tests the network transfer rate with the external server was measured to be 10 MB/s. The HyperLogLog sketch is designed for streams of data, and we can take advantage of this property to com- pose the cardinality estimation capabilities with other pipelines. Here, the overhead of the cardinality counting is minimal with respect to I/O: downloading SRR1304364 and piping it into our HLL implementation adds only 1% to the overall runtime ( 199.7 ± 9.5 seconds for counting and saving, versus 199.5±13.6 seconds for simply saving the ﬁle). Thus our HLL implementation can be used "mid- stream" to evaluate the effects of streaming lossy compres- sion and error trimming. Competing interests The authors have no conﬂicts of interest or competing in- terests to disclose. Benchmarking n = f (x) ∧(m −1) All tests were executed on a server hosted by Rackspace. This machine has an Intel Xeon E5-2680 CPU with 20 physical cores (2 sockets) running at 2.80GHz, 60 GB of The two basic operations of a sketch are Add (or update) and Merge. Adding an element x involves calculating its Page 2 of 5 Page 2 of 5 F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint Comparison with exact counters To test the correctness of the implementation we used two exact cardinality data structures for comparison to our HLL sketch implementation. For exact cardinality we created a Python implementation using the standard li- brary (“collections.Counter”) and another in C++ using the Google sparsehash library. Neither are parallelized. Discussion/Conclusions Both implementations are impractical for large cardinal- ities, so we chose two relatively small datasets from the Sequence Read Archive for benchmarking: We present an open and remixable implementation of the HyperLogLog sketch for cardinality counting of DNA k- mers, written in C++. The implementation scales well to multiple threads, and uses OpenMP for task coordination. • SRR797943, containing 417 PacBio and 454 se- quences with average length 1690 basepairs and to- taling 704,951 basepairs, with 670,487 unique k- mers. Referred to below as the small dataset, Scaling behavior W h l d Noga Alon, Yossi Matias, and Mario Szegedy. The space complex- ity of approximating the frequency moments. In Proceedings of the twenty-eighth annual ACM symposium on Theory of computing, pages 20–29. ACM, 1996. URL http://dl. acm.org/citation.cfm?id=237823. We chose a larger dataset for examining the scaling per- formance of our HLL implementation. This larger dataset, SRR1304364, contains 163,379 PacBio sequences with average length 12,934 bp, and 2,113,086,496 basepairs in total. We examined how our implementation scaled with num- ber of threads. Since hashing is CPU-bound, the prob- lem can be easily parallelized. We ran a simple bench- mark to discover the I/O lower bound, using the same input and read parsing infrastructure as the HLL sketch tests, but without performing any kind of processing. Fig- ure 1 shows the results of these tests, where we found 16 threads are needed to saturate I/O on this particular setup, which has 16 physical cores. Páll Melsted and Bjarni V. Halldórsson. Kmerstream: streaming algorithms for k-mer abundance estima- tion. Bioinformatics, 30(24):3541–3547, 2014. doi: 10.1093/bioinformatics/btu713. URL http: //bioinformatics.oxfordjournals.org/content/ 30/24/3541.abstract. Jason Pell, Arend Hintze, Rosangela Canino-Koning, Adina Howe, James M Tiedje, and C Titus Brown. Scaling metagenome sequence assembly with probabilistic de bruijn graphs. Proceedings of the National Academy of Sciences, 109(33):13272–13277, 2012. Page 3 of 5 Page 3 of 5 F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint Philippe Flajolet, Éric Fusy, Olivier Gandouet, and Frédéric Me- unier. Hyperloglog: the analysis of a near-optimal cardinality estimation algorithm. DMTCS Proceedings, (1), 2008. Daniel R. Zerbino and Ewan Birney. Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome research, 18(5):821–829, 2008. Qingpeng Zhang, Jason Pell, Rosangela Canino-Koning, Ad- ina Chuang Howe, and C. Titus Brown. F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint Scaling behavior W h l d These are not the k- mers you are looking for: Efﬁcient online k-mer counting us- ing a probabilistic data structure. PLoS ONE, 9(7):e101271, 07 2014. doi: 10.1371/journal.pone.0101271. URL http: //dx.doi.org/10.1371%2Fjournal.pone.0101271. Michael R. Crusoe, Hussien F. Alameldin, Sherine Awad, Elmar Bucher, Adam Caldwell, Reed Cartwright, Amanda Charbon- neau, Bede Constantinides, Greg Edvenson, Scott Fay, Jacob Fenton, Thomas Fenzl, Jordan Fish, Leonor Garcia-Gutierrez, Phillip Garland, Jonathan Gluck, Iván González, Sarah Guer- mond, Jiarong Guo, Aditi Gupta, Joshua R. Herr, Adina Howe, Alex Hyer, Andreas Härpfer, Luiz Irber, Rhys Kidd, David Lin, Justin Lippi, Tamer Mansour, Pamela McA’Nulty, Eric McDonald, Jessica Mizzi, Kevin D. Murray, Joshua R. Nahum, Kaben Nanlohy, Alexander Johan Nederbragt, Hum- berto Ortiz-Zuazaga, Jeramia Ory, Jason Pell, Charles Pepe- Ranney, Zachary N Russ, Erich Schwarz, Camille Scott, Josiah Seaman, Scott Sievert, Jared Simpson, Connor T. Skennerton, James Spencer, Ramakrishnan Srinivasan, Daniel Standage, James A. Stapleton, Joe Stein, Susan R Steinman, Benjamin Taylor, Will Trimble, Heather L. Wiencko, Michael Wright, Brian Wyss, Qingpeng Zhang, en zyme, and C. Titus Brown. Evangelos Georganas, Aydin Buluç, Jarrod Chapman, Leonid Oliker, Daniel Rokhsar, and Katherine Yelick. Parallel de bruijn graph construction and traversal for de novo genome assem- bly. In High Performance Computing, Networking, Storage and Analysis, SC14: International Conference for, pages 437– 448. IEEE, 2014. Yousra Chabchoub and Georges Hébrail. Sliding hyperloglog: Estimating cardinality in a data stream over a sliding win- dow. In Data Mining Workshops (ICDMW), 2010 IEEE International Conference on, pages 1297–1303. IEEE, 2010. Stefan Heule, Marc Nunkesser, and Alexander Hall. Hyperloglog in practice: algorithmic engineering of a state of the art car- dinality estimation algorithm. In Proceedings of the 16th International Conference on Extending Database Technology, pages 683–692. ACM, 2013. Page 4 of 5 Page 4 of 5 F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. ; https://doi.org/10.1101/056846 doi: bioRxiv preprint F1000Research 2014 - DRAFT ARTICLE (PRE-SUBMISSION) . CC-BY 4.0 International license available under a not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint (which was this version posted June 7, 2016. Scaling behavior W h l d ; https://doi.org/10.1101/056846 doi: bioRxiv preprint 1 2 4 8 16 threads 32 64 128 256 512 1024 Elapsed (wall clock) time (seconds) Walltime (s), CI 95% Parallel HLL Just I/O Figure 1. Walltime and lower bound(I/O) Implementation Time (seconds) Memory (MB) Cardinality Error HLL 0.193 13.44 670,328 0.0002 C++/sparsehash 3.888 77.64 670,487 0 Python 4.306 83.48 670,487 0 Table 1. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the small size dataset Implementation Time (seconds) Memory (MB) Cardinality Error HLL 12.08 13.38 17,686,322 1.01 C++/sparsehash 344.04 2,018.50 17,510,301 0 Python 891.17 2,056.05 17,510,301 0 Table 2. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the medium size dataset 1 2 4 8 16 threads 32 64 128 256 512 1024 Elapsed (wall clock) time (seconds) Walltime (s), CI 95% Parallel HLL Just I/O Figure 1. Walltime and lower bound(I/O) Table 1. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the small size dataset Implementation Time (seconds) Memory (MB) Cardinality Error HLL 12.08 13.38 17,686,322 1.01 C++/sparsehash 344.04 2,018.50 17,510,301 0 Python 891.17 2,056.05 17,510,301 0 Table 2. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the medium size dataset time and memory consumption for HLL and two exact cardinality implementations (Python sh) using the small size dataset Table 2. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the medium size dataset Table 2. Wall clock time and memory consumption for HLL and two exact cardinality implementations (Python and C++/sparsehash) using the medium size dataset Page 5 of 5 Page 5 of 5	47,513
2004ENAM0034_2	French-Science-Pile	Open Science	Various open science	2,004	Management des risques, stratégies d'entreprise et reglementation : le cas de l'industrie maritime	None	French	Spoken	6,971	10,637	Il est possible de tirer plusieurs conclusions de ces critiques. Tout d’abord, il convient de ne pas considérer que les analyses techniques sont le seul instrument dont nous disposons pour évaluer les risques. Fort heureusement, l’apport d’autres disciplines peut permettre de les enrichir avec profit : les analyses techniques ne possèdent qu’une validité locale, qu’il convient d’intégrer dans une approche plus générale. A un deuxième niveau, une des conditions de succès et de crédibilité de l’analyse de risques est sans doute le respect par l’analyste d’une certaine éthique, face notamment à l’augmentation des puissances de calcul informatique, qui fait que les obstacles techniques sont devenus bien souvent beaucoup moins insurmontables qu’ils ne l’étaient il y a quelques années de cela. Ahearne (1999) présente ainsi une série de recommandations qui s’apparentent pour partie à du bon sens14, mais permettent de délimiter ce que pourraient être de bonnes pratiques pour une analyse de risques consciente de ses limites et de ses faiblesses. 13 Nous reviendrons sur ce point plus en détail dans la section 1.3. Ahearne (1999) cite ainsi l’importance de la vérification mathématique, de la rigueur scientifique pour les aspects touchant à la modélisation des effets d’un événement redouté, de la démonstration des limites de l’analyse, de la clarté des résultats, de l’intégrité vis-à-vis des donneurs d’ordre, même lorsque les résultats apparaissent contraire aux attentes, et de l’implication du public. - Après avoir présenté les outils de l’analyse de risque et les critiques qui peuvent leur être faites15, il importe maintenant de nous interroger sur leur finalité managériale, ou plus généralement sur ce qui en fait autant d’outils de gestion, au sens de Moisdon (1997)16. Nous allons ici défendre l’idée que les outils d’analyse de risques s’inscrivent dans une triple logique : celle de l’aide à la décision, celle de la légitimation (qui recouvre une double dimension de représentation et de communication), et celle de l’apprentissage17. L’aide à la décision est bien entendu le rôle premier usuellement assigné à l’analyse de risques. Le processus que nous avons décrit se complète généralement par une évaluation technico-économique des mesures de réduction du risque à implémenter, souvent à l’aide d’indicateurs issus d’une analyse coûtbénéfice ou coût-efficacité (Swaney, 1997)18. Au-delà de cette application un peu mécaniste des techniques d’analyse de risques, celles-ci peuvent être considérées comme de véritables outils de management, dans la mesure où elles rendent nécessaire l’établissement de priorités et s’inscrivent ainsi dans une démarche générale d’allocation raisonnée des ressources. Elles peuvent ainsi servir de fondement à des décisions de modifications d’arrangements organisationnels, 15 Il va de soi que cette présentation n’a pu être que succincte. Pour plus de détails sur les méthodes elles-mêmes, nous renvoyons le lecteur aux ouvrages de référence de Henley et Kumamoto (1996) et de Villemeur (1988). 16 Pour Moisdon (1997, p. 7), un outil de gestion se définit comme un « ensemble de raisonnements et de connaissances reliant de façon formelle un certain nombre de variables issues de l’organisation, qu’il s’agisse de quantités, de prix, de niveaux de qualité ou de tout autre paramètre, et destiné à instruire les divers actes classiques de la gestion, que l’on peut regrouper dans les termes de la trilogie classique : prévoir, décider, contrôler ». 17 Bien souvent, seules au mieux les deux premières dimensions sont mises en avant, en particulier chez de nombreux praticiens ou chercheurs issus des sciences de l’ingénieur, même si in fine, la production de connaissances et l’apprentissage apparaissent comme un résultat logique de l’analyse. Cf. par exemple Paté-Cornell (2002), p. 320 : « Engineering risk analysis is performed for two reasons: to show that a system is safe enough and to set priorities in risk management (...) », et p. 333 « What matters is that the risk analysis process allows discovering system weaknesses that were not anticipated and were not necessarily part of the motivation of the risk analysis. » Il est intéressant toutefois de constater d’une part que ce n’est qu’au terme de son article que Paté-Cornell arrive à cette conclusion, et de l’autre que la nature de la connaissance produite reste instrumentale. Pour Paté-Cornell (2002), les nouvelles connaissances auxquelles conduit l’analyse de risques sont celles de l’analyste (ou du manager) sur l’organisation, non de l’organisation sur elle-même : il n’est donc pas question ici d’apprentissage organisationnel, contrairement à ce que nous verrons plus loin. 33 - comme l’explique par exemple Paté-Cornell (1996). Loin d’être confinée aux techniques de l’ingénieur, l’analyse de risques peut donc se faire le support de décisions d’essence managériale, et non seulement techniques19. Cet outil de décision ne peut cependant être considéré in abstracto. Comme cela a été écrit ailleurs (Hatchuel et Moisdon, 1997), un modèle d’aide à la décision n’est en effet rarement que cela. En pratique, tout modèle d’aide à la décision20 s’insère dans des dispositifs organisationnels préexistants, et les conclusions auxquels il permet d’aboutir ont des conséquences qui font qu’il ne peut être décontextualisé. Dans cette perspective, l’analyse de risques sert tout autant d’outil d’aide à la décision que d’outil de légitimation d’une décision, et ce à au moins deux titres. Elle permet tout d’abord de systématiser à l’aide d’outils relativement simples un état de connaissance du système modélisé. Par suite, elle peut constituer un appui à la décision, et permettre de la justifier. Ce type d’emploi se retrouve notamment dans certaines applications réglementaires de l’analyse de risques, comme nous le verrons dans le dernier chapitre de la thèse. Le versant négatif de cet usage en termes de légitimation de la décision correspond à son utilisation de manière ad hoc. En effet, la complexification des systèmes industriels rend bien souvent, et pour partie paradoxalement, le recours à des calculs probabilistes plus difficile : paradoxalement, car l’on pourrait s’attendre à ce qu’au contraire l’importance des incertitudes en jeu pousserait à vouloir les prendre en compte. En réalité, elle conduit au contraire fréquemment les ingénieurs de conception à se raccrocher aux codes déterministes, par défaut de compréhension du fonctionnement du système. Par suite, le fait que les représentations (graphiques ou autres) que l’analyse de risques emploie en fassent un puissant outil de communication et la relative simplicité d’utilisation des 18 Nous reviendrons plus avant sur les techniques d’évaluation et leurs limites lors de notre chapitre six. 19 Nous développerons l’apport de l’équipe du Professeur Paté-Cornell et le lien entre analyse de risques et analyse de la décision dans la section 1.3.1. 20 Hatchuel et Moisdon (1997) partent de l’exemple de l’usage des techniques d’analyse de la décision chez Total et chez Elf. 34 - techniques qui la composent peuvent conduire à un détournement de son usage dans une perspective de rationalisation a posteriori21. L’analyse de risques ne se réduit cependant pas à cette vision négative. Bien au contraire, les mécanismes que nous venons de mettre en évidence permettre d’en comprendre l’usage comme vecteur d’apprentissage. En ce qu’elle se fonde justement sur une représentation du système industriel qu’elle étudie, l’analyse de risques conduit en effet à en questionner les rouages, et par voie de conséquence, offre la possibilité, non seulement d’en améliorer le fonctionnement, mais aussi et surtout d’acquérir des connaissances collectives nouvelles, dans un mécanisme proche de celui décrit par Hatchuel (1994) dans son étude des activités de conception. C’est notamment en cela qu’une décentralisation du processus telle que celle que prônent Mays et Poumadère (1989) et qui passe par la participation des opérateurs à l’analyse peut engendrer un apprentissage collectif et de meilleures pratiques. Dans cette perspective, on notera qu’il n’est presque pas indispensable que l’analyse soit menée à son terme : le processus même de participation peut suffire à ce qu’émergent des connaissances collectives. Cette dernière thèse n’a malheureusement été jusqu’à présent que fort peu creusée, alors qu’elle est sans doute une des clés de l’acceptation de l’analyse de risques comme réel outil de gestion, en même temps qu’elle peut contribuer à asseoir la légitimité de ses résultats. 1.2. La prise en compte du « facteur humain » Cette dernière dimension de l’analyse de risques comme vecteur d’apprentissage nous permet une transition vers l’étude de la prise en compte des individus dans l’analyse, que nous avons jusqu’à présent laissée de côté au profit 21 Ce point n’a jusqu’à présent été que peu étudié à notre connaissance, si l’on excepte quelques pages dans Perrow (1984, pp. 307 sqq.) et l’article de Webler et al. (1992), qui analysent dans une optique théorique-critique ce phénomène à une échelle plus générale, en se fondant sur les travaux de l’Ecole de Francfort, et notamment ceux d’Habermas. Les considérations qui précèdent 35 - des systèmes techniques. Or, dès les tous débuts de l’analyse de risques, les ingénieurs se sont efforcés d’intégrer les actions des opérateurs dans leurs évaluations par ce qu’ils ont qualifié de « facteur humain ». Ainsi, les origines de la méthode THERP (sur laquelle nous reviendrons plus loin) au début des années 1960 sont contemporaines des développements des arbres de défaillances. La démarche générale d’analyse du facteur humain consiste à étudier les tâches que les opérateurs doivent accomplir, pour en déduire les erreurs qu’ils sont susceptibles de commettre et quantifier leur probabilité d’occurrence, et ce afin de pouvoir éventuellement parer à ces erreurs. Une telle analyse de la fiabilité humaine s’interface naturellement avec l’analyse des risques associés à des systèmes techniques, faisant de l’élément humain un des facteurs pour lesquels des mesures de réduction du risque peuvent être envisagées, qu’il s’agisse de l’introduction de dispositifs tenant à l’interface homme-machine (conception et ergonomie du poste de travail, nature des alarmes en cas de danger...), à la formation, à l’organisation des équipes et au temps de travail, ou encore à l’automation de certaines tâches. Nous allons maintenant passer rapidement en revue les principales méthodes existantes dans ce domaine, en nous intéressant successivement à l’analyse des tâches et à l’analyse de la fiabilité humaine. Dans un troisième temps, nous présenterons quelques critiques des méthodes existantes et de possibles voies d’amélioration. 1.2.1. L’analyse de tâches L’analyse des tâches tout d’abord a pour but de dresser un tableau aussi exhaustif que possible des interactions homme-machine et homme-homme. Elle se définit comme l’étude de ce qu’un opérateur (ou une équipe d’opérateurs) doit effectuer en termes d’actions et/ou de processus cognitifs en vue d’atteindre les reflètent quant à elles largement notre propre expérience de recherche-action dans des sociétés d’ingénierie. 36 - objectifs du système (Sheridan, 1999). Elle peut être traitée de multiples manières et recouvre de nombreuses techniques, suivant le niveau de détail souhaité et l’objectif poursuivi : Kirwan et Ainsworth (1992), dans leur ouvrage de référence en la matière, ne recensent pas moins de 41 méthodes22, qui touchent tant à la collecte des données qu’à l’organisation et à la représentation de celles-ci. La collecte des données tout d’abord peut prendre deux formes essentielles. L’observation directe du comportement de l’opérateur est sans doute la manière la plus immédiate et intuitive d’accéder à la connaissance des activités mises en œuvre par ce dernier. Elle peut prendre place sur le lieu de travail de l’opérateur (ce qui permettra de disposer d’informations précieuses sur les conditions environnementales) ou dans un simulateur reproduisant les principaux éléments du poste de travail. Plusieurs techniques peuvent être mises en œuvre (Drury, 1995), de la simple prise de note par l’observateur (qui peut, suivant les cas, être dissimulé en utilisant un dispositif vidéo ou non) à l’utilisation de protocoles verbaux (Bainbridge et Sanderson, 1995), durant lesquels l’opérateur verbalise la tâche qu’il est en train de réaliser, en passant par l’échantillonnage de l’activité et la mesure des temps, ou encore la participation active de l’observateur au travail, par exemple dans le cas d’un travail en équipe. L’intérêt de telles approches est bien évidemment la richesse des informations collectées, qui en est aussi une limite en termes de lourdeur de traitement. Cependant, leur principale difficulté reste le phénomène de « l’effet Hawthorne » (Mayo, 1933) de perturbation de l’objet observé par l’observateur qui est ici inévitable, sauf à adopter des méthodes de dissimulation totale, discutables sur un plan déontologique. La deuxième voie d’accès aux données nécessaires à l’analyse des tâches consiste à interroger l’opérateur en différé (Sinclair, 1995), grâce à des entretiens 22 Kirwan et Ainsworth (1992) étudient plus particulièrement 25 techniques, dont certaines toutefois sont directement empruntées à d’autres domaines (tels que par exemple l’analyse par arbre de défaillances ou l’HAZOP appliquée à l’analyse de l’erreur humaine) ou sont si 37 - plus ou moins structurés (Meister, 1985) ou à des questionnaires (Oppenheim, 1992). Ces techniques de collecte présentent l’avantage d’être potentiellement moins intrusives que l’observation en ce qu’elles sont déconnectées du lieu de travail. Elles restent cependant soumises à de possibles biais dont l’analyste doit s’efforcer de se prémunir. Les données collectées vont ensuite être devoir être traitées et mises en forme, essentiellement sous la forme d’analyses tabulaires, hiérarchiques et temporelles (timeline analysis). L’analyse tabulaire de tâches repose sur une décomposition des tâches suivant les aspects auxquels l’analyste s’intéresse. En ce sens, elle est assez proche de l’AMDEC. Les résultats de la décomposition sont ensuite enregistrés dans un tableau dont les colonnes correspondent aux catégories jugées dignes d’intérêt, telles que, par exemple, la phase durant laquelle la tâche a lieu, le code de la tâche, son but, les conditions dans lesquelles elle se déroule et qui sont susceptibles d’avoir un impact sur la performance de l’opérateur, la fonction de l’opérateur en charge de la tâche, l’information dont il dispose, l’action requise, l’existence éventuelle de procédures, le feedback qu’il est susceptible de recevoir, les communications qui peuvent avoir lieu, et les erreurs éventuelles liées à la tâche (Vivalda, 1998)23. Les catégories finales sont à choisir en fonction des tâches à analyser et de l’objectif poursuivi. A ce titre, les résultats de questionnaires préliminaires peuvent s’avérer particulièrement utiles. L’analyse hiérarchique de tâches est une présentation sous forme d’arbre des tâches et sous-tâches qu’un opérateur doit effectuer pour accomplir une activité. Dans un second temps, l’arbre est généralement transformé en tableau pour en faciliter la lisibilité. Le déroulement de l’analyse commence par l’examen d’un objectif à atteindre et du plan des tâches à mettre en œuvre pour atteindre cet étroitement liées à des techniques d’analyse de l’erreur humaine qu’il est difficile de les en distinguer. 23 D’autres catégories sont bien entendues utilisables, telles que celles décrites par Kirwan et Ainsworth (1992). 38 - objectif (c’est-à-dire l’ordre et les conditions dans lesquels les tâches doivent être effectuées). Les tâches identifiées sont ensuite décomposées en sous-tâches puis en opérations élémentaires jusqu’à ce que la description obtenue réponde aux besoins de l’analyse. Une règle d’arrêt intuitive pour la décomposition consiste à considérer de manière approximative la probabilité pour que l’opérateur ne réussisse pas à mettre en œuvre l’opération considérée avec succès, et les conséquences de cet échec. Lorsque le niveau atteint est suffisamment détaillé pour que les conséquences d’une erreur ou la possibilité d’échec soient faibles, l’analyse peut être considérée comme complète pour la hiérarchie considérée. L’analyse temporelle enfin est une technique qui vise à saisir les contraintes temporelles qui pèsent sur l’opérateur, et elle est en cela l’héritière des techniques tayloriennes. Elle se caractérise plus par ses principes que par une procédure rigoureuse, et conduit à présenter le déroulement de l’activité de l’opérateur sous la forme d’un graphe qui distingue le temps de l’action de l’opérateur de ses pauses et des temps de réponse du système. 1.2.2. L’analyse de la fiabilité humaine : de THERP à CREAM Une fois que le comportement de l’opérateur a été précisément décrit, il devient possible de procéder à une analyse de la fiabilité humaine, qui vise à estimer une probabilité de défaillance. La jungle des méthodes qui permettent d’aboutir à ce but est particulièrement dense, en raison des difficultés à saisir la spécificité et la richesse du comportement humain : certaines d'entre elles vont se focaliser sur les interactions hommes-machines, d'autres sur les aspects organisationnels, d'autres encore sur les déterminants cognitifs du comportement, etc. Avant de présenter les plus importantes des nombreuses méthodes d’analyse de la fiabilité humaine24, il nous paraît nécessaire d’expliquer les raisons 24 Kirwan (1994) et Hollnagel (1998) proposent une analyse quasi-exhaustive des méthodes existantes, hors évidemment les méthodes ad hoc. Reason (1997) reprend certaines d’entre elles afin d’examiner en quoi elles tiennent compte de l’organisation dans laquelle l’opérateur agit. Moieni et al. (1994a et 1994b) présentent quant à eux les aspects plus techniques liés à la mise en œuvre de quelques méthodes développées dans les années 1980. 39 - principales qui semblent être à l'origine de leur diversité ; ces raisons vont en effet permettre de mieux comprendre les critiques que nous formulons à leur encontre à l’issue de cette section. La première raison repose sur le fait qu’à l’inverse de la modélisation des aspects techniques du fonctionnement d'un système dans l’analyse de risques qui, sans ambiguïté, s’appuie sur des outils issus d’une culture de l'ingénieur, ce qui a permis une sédimentation progressive et une comparaison univoque des mérites respectifs des méthodes proposées successivement pour aboutir à un relatif consensus sur les meilleures d’entre elles25, de nombreuses disciplines se préoccupent du comportement humain, de l'ergonomie à la théorie des organisations, en passant par la psychologie (cognitive et sociale), l'ingénierie des systèmes complexes, la sociologie, l'intelligence artificielle... Les méthodes que ces disciplines proposent ne sont pas commensurables, en ce qu'elles conduisent à étudier une seule facette du comportement humain, qui en comporte de nombreuses. La pertinence d'une approche ne peut donc être appréhendée que localement, vis-à-vis d'une étude particulière. Il n'existe pas d'approche générique, comme c'est le cas pour l'étude des systèmes techniques où des méthodes telles que par exemple l'AMDEC, l'HAZOP, les arbres de défaillances..., ont fait leurs preuves, ou sont tout au moins acceptées par la communauté des ingénieurs comme ayant un degré de validité suffisant pour que les résultats auxquels elles conduisent soit eux-mêmes jugés acceptables, pour autant qu’elles aient été correctement appliquées. Le second facteur qui permet de comprendre la diversité des méthodes utilisées en analyse de la fiabilité humaine est le fait qu’à l’inverse de la plupart des techniques d'ingénierie qui permettent d'aboutir, de manière plus ou moins aisée, à une mesure de performance du système, la diversité des facteurs qui 25 Certaines études tendent à tempérer cette vision, telles que par exemple celles de Rouvroye et Brombacher (1999) et Rouvroye et van den Bliek (2002), qui montrent que l’application de plusieurs techniques d’ingénierie à l’évaluation de risques semblables conduit à des résultats différents. Les auteurs proposent l’utilisation d’une « analyse de Markov étendue » pour compenser cette difficulté. On notera cependant que les différences ne sont pas si importantes que cela (souvent moins d’un ordre de magnitude) et peuvent être prises en compte par un usage approprié de l’analyse de sensibilité. D’autre part, il n’en reste pas moins que les quelques six méthodes présentées dans ces articles font l’objet d’un relatif consensus, ce qui n’est certainement 40 - président au comportement humain est beaucoup plus importante, ce qui rend la mesure beaucoup plus difficile (Mosneron-Dupin, 1994). Ceci a deux types de conséquences quant à la prolifération des méthodes : d’une part, il est difficile de trouver un critère clair de validité, et donc d’éliminer les méthodes les moins performantes ; d’autre part, nombre de méthodes ne sont en réalité que des applications de techniques de recueil d’avis d’expert à l’évaluation de la fiabilité humaine26. Malgré cette diversité et cette difficulté à comparer les méthodes d'analyse de la fiabilité humaine entre elles, il semble cependant que certaines se dégagent du lot. En suivant une taxonomie aujourd’hui largement établie, il est courant de distinguer deux générations de méthodes, la frontière entre ces générations se situant au tournant des années 1990. Nous allons nous intéresser dans ce qui suit à deux méthodes, qui sont respectivement représentatives de la première et de la deuxième génération : nous commencerons ainsi par décrire THERP (Swain et Guttman, 1983) puis, dans un second temps, la méthode CREAM (Cognitive Reliability and Error analysis Method, Hollnagel, 1998). Ces méthodes présentent ceci d’intéressant qu’à un titre ou un autre, elles sont exemplaires des techniques utilisées en analyse du facteur humain. Elles vont également nous permettre d’en montrer les limites dans l’appréhension des risques. La méthode THERP est sans doute celle qui exemplifie le mieux les méthodes classiques de première génération27. Elle trouve son origine au début des années 1960 et reste largement une méthode de référence. Les hypothèses pas le cas des méthodes d’analyse de la fiabilité humaine, beaucoup plus nombreuses : Hollnagel (1998) avance ainsi le chiffre de 35 à 40 méthodes. 26 Les méthodes HEART (Human Error Assessment and Reduction Technique), JHEDI (Justification of Human Error Data Information), SLIM-MAUD (Success Likelihood Methodology using Multi Attribute Utility Decomposition) que nous ne détaillerons pas plus avant mais qui sont décrites en détail par exemple dans Kirwan (1994) rentrent par exemple dans cette catégorie. Parmi les méthodes de première génération les plus connues, on peut citer, outre THERP, la méta-méthode SHARP (Systematic Human Action Reliability Procedure), qui intègre des critères de sélection de méthode d’analyse de la fiabilité humaine, la méthode SHERPA (Systematic Human Error Reduction and Prediction Approach), la méthode des matrices de confusion, et la 41 - sous-jacentes à THERP (et à la plupart des méthodes de première génération) sont que les individus doivent être considérés pratiquement au même niveau que les autres éléments d'un système technique, à la seule réserve peut-être qu'ils sont influencés par les conditions dans lesquelles ils opèrent et qu’ils disposent d'une forme de redondance intrinsèque : s'ils commettent une erreur ou une action susceptible d'avoir des conséquences néfastes, ils peuvent effectuer une récupération et gérer le danger spontanément. Les humains peuvent être caractérisés par un taux d'erreur a priori, de même que les machines peuvent être caractérisées par un taux de défaillance. Dans le cadre de THERP, les causes de l'erreur sont largement laissées en retrait ; toutefois, on considère que le taux d'erreur peut être ajusté par des facteurs de performance (Performance Shaping Factors) internes (l’intelligence de l’opérateur, son expérience, sa condition physique...), externes (caractéristiques du poste de travail, bruit, mais aussi structure organisationnelle), stresseurs (facteurs qui influencent le stress psychologique (rapidité de la tâche, longueur des pauses...) ou physiques (fatigue, faim, soif...)), en utilisant une équation linéaire28. La méthode THERP est sans doute la plus complète de toutes les méthodes d’analyse de la fiabilité humaine, tout d’abord en ce qu’elle retrace l’ensemble de la démarche, y compris l’analyse de tâches, ensuite en ce qu’elle spécifie également la manière d’utiliser des arbres proches des arbres d’événements pour modéliser les actions de l’opérateur ainsi que la possibilité de récupération en cas d’erreur, enfin parce qu’elle comporte29 des tables de probabilités a priori d’erreurs, largement utilisées (même si c’est parfois de manière abusive) par nombre de praticiens de l’analyse de la fiabilité humaine. méthode OAT (Operator Action Tree). Hollnagel (1998) et Kirwan (1994) décrivent ces méthodes en détail. 28 n PE = HEPE ∑ PSFk .W k + C , avec PE la probabil ité d ’action erronée , HEPE la probabilité a priori k=1 € d’action erronée, PSFk la va leur du Performance Shaping Factor k, Wk le po ids associé à ce Performance Sha ping Factor , n le nombre total de Performance Shaping Factors, et C une constante numérique. 29 Swain et Guttmann (1983), chapitre 20. 42 - La seconde génération des méthodes d’analyse de la fiabilité30 repose à l'inverse sur la prise en compte de la dimension cognitive contextualisée de l’action humaine (Hollnagel, 1993), et sépare clairement le phénomène de l'action erronée de la défaillance sous-jacente, alors que la plupart des approches avaient précédemment tendance à opérer une confusion entre ces deux phénomènes, les regroupant sous la catégorie générique de « l’erreur humaine » (Dougherty, 1995). En cela, la prise en compte des phénomènes cognitifs s’inscrit en réaction au modèle mécaniste31 de l’opérateur, sous-jacent (ou explicite) dans les méthodes de première génération (Hollnagel, 1996). Les défaillances cognitives, dans cette approche, sont à la source des actions erronées, elle-même comprises comme des actions ayant des conséquences non désirables32. La méthode CREAM (Hollnagel, 1998) est sans doute aujourd’hui l’un des efforts les plus aboutis en matière d’analyse de fiabilité humaine de deuxième génération, même si elle est loin de s’être autant répandue que THERP. Elle s’appuie sur le modèle COCOM (COntextual COntrol Model) de l’opérateur (Hollnagel, 1993), qui distingue d’une part quatre « modes de contrôle » de l’action33 et d’autre part quatre types de « compétences » qui correspondent à des fonctions cognitives de base (observation, interprétation, planification, exécution). La réalisation de ces fonctions, et donc la fiabilité humaine, va dépendre des conditions dans lesquelles elles se déroulent, qui peuvent être regroupées dans ce 30 Parmi les méthodes de seconde génération qui prennent en compte les facteurs cognitifs, on peut notamment citer, outre CREAM, les méthodes Intent, COGENT (Cognitive Event Tree System, Gertman, 1993), ATHEANA (A Technique for Human Error ANAlysis, Dougherty, 1998). 31 Ce modèle de l’opérateur repose généralement sur deux piliers : une appréhension du comportement comme réponse à un stimulus et une décroissance de la fiabilité humaine au cours du temps. 32 A l’inverse, dans les méthodes de première génération, le fonctionnement de l’opérateur est largement vu comme une « boîte noire » dont on ne connaît que les probabilités d’erreur a priori. 33 Par ordre croissant de fiabilité : le mode « brouillé » (scrambled), dans lequel l’action de l’opérateur résulte du hasard, ce qui est le cas lorsque l’action est complexe, se déroule dans des conditions difficiles... ; le mode « opportuniste » dans lequel l’action de l’opérateur résulte largement des caractéristiques saillantes du contexte, et des solutions les plus immédiates qui se présentent ; le contrôle « tactique » repose quand à lui sur une certaine planification de l’action, qui repose autant que faire se peut sur les procédures prescrites ; le mode de contrôle « stratégique » enfin correspond à une planification avancée de la tâche à accomplir et à une prise en compte de ses conséquences. 43 - qu’Hollnagel appelle des conditions communes de performance, qui ont trait à l’adéquation des dispositifs organisationnels, aux conditions de travail, à l’adéquation des interfaces homme-machine, à la disponibilité de procédures ou de plans, au nombre de buts simultanés à atteindre, au temps disponible, à l’heure d’exécution de la tâche et au respect des rythmes circadiens, à l’adéquation de la formation et de l’expérience et à la qualité de la collaboration au sein de l’équipe. Les modalités que prendront ces facteurs vont influencer la performance de l’opérateur par le biais de l’impact qu’elles vont avoir sur l’exécution des fonctions cognitives de base. Hollnagel propose plusieurs applications de son cadre méthodologique à l’analyse rétrospective d’accidents, à l’analyse prédictive qualitative de la performance, et à l’analyse prédictive quantitative de la performance. C’est sur ces deux dernières applications que nous allons nous arrêter. L’analyse qualitative que décrit Hollnagel est particulièrement intéressante en ce qu’elle ne repose pas sur un modèle linéaire. Au contraire, elle s’appuie sur l’existence de bouclages entre les différentes fonctions (une mauvaise observation peut mener à une interprétation biaisée, engendrant elle-même une erreur d’exécution, qui à son tour peut perturber le système de telle sorte qu’une nouvelle interprétation erronée peut avoir lieu et ainsi de suite), et ainsi sur une dynamique complexe. Par suite, il s’agit de comprendre la séquentialité de l’enchaînement des fonctions pour saisir la propagation des facteurs qui vont influencer la performance. On est donc loin ici d’une vision mécaniste linéaire réductrice du fonctionnement de l’opérateur. En revanche, la quantification passe par une simplification du modèle. Hollnagel propose deux méthodes pour y parvenir. La méthode simplifiée vise à établir le mode de contrôle (brouillé, opportuniste, tactique ou stratégique) de la tâche à partir d’une somme de scores associés aux facteurs communs de performance. A partir de là, Hollnagel (1998) fournit une table d’intervalles de probabilités de défaillance associés à chaque mode de contrôle, qu’il est possible d’utiliser comme approximation de la probabilité d’échec de la tâche. La méthode étendue quant à elle se distingue de la précédente en ce qu’elle prescrit d’opérer un profil de « demande cognitive » de chaque tâche. Ce profil s’obtient en 44 - décomposant les tâches en activités cognitives élémentaires34, que l’on regroupe ensuite dans les fonctions cognitives de base (interpréter, observer, planifier, exécuter) à l’aide d’une matrice. Ceci permet d’aboutir à un graphe qui associe à chaque tâche les fonctions cognitives en jeu, qui peuvent avoir une importance inégale (par exemple, une tâche peut impliquer majoritairement de l’observation, mais aussi un peu d’exécution et de planification). Dans un second temps, l’analyste devra identifier les défaillances cognitives les plus vraisemblables, pour lesquelles Hollnagel (1998) fournit des probabilités tirées de la littérature, et qui seront pondérées, soit par un facteur dépendant du mode de contrôle, soit par un facteur dépendant des facteurs communs de performance en jeu, pour aboutir à une probabilité d’échec de la tâche considérée. 1.2.3. La nécessité d’un renouvellement des méthodes Après ces rapides présentations, nous allons maintenant examiner quelques-unes des critiques portées sur les méthodes d’analyse de la fiabilité humaine. Les critiques de la méthode THERP tout d’abord sont nombreuses, et nous allons ici reprendre les principales. Paradoxalement, c’est de son principal architecte que viennent les attaques les plus virulentes (Swain, 1990)35, parmi lesquelles on relève le fait que les données concernant la fiabilité humaine sont en général de faible qualité (voir aussi Sträter et Bubb, 1999 sur ce point), qu’il n’existe pas vraiment d’accord sur les méthodes d’encodage de jugement d’expert (cf. aussi supra, notre évocation de cette question), que les simulations sont difficiles à calibrer, qu’il n’existe quasiment pas de validation empirique des méthodes36, qu’elles n’ont pas de réalisme psychologique, et que le traitement des 34 Coordonner, communiquer, comparer, diagnostiquer, évaluer, exécuter, identifier, maintenir, surveiller, observer, planifier, enregistrer, réguler, scanner du regard, vérifier. 35 Swain (1990) en arrive même à écrire : « At the present time, fully adequate HRAs have not been performed ». Ce constat de 1990 semble malheureusement être encore d’actualité aujourd’hui. 36 On relèvera cependant l’effort d’évaluation des méthodes HEART, THERP et JHEDI à l’université de Birmingham (Basra et Kirwan, 1998), puis, 5 ans plus tard, par le Nuclear Industry Management Committee du Royaume-Uni (Kennedy et al., 2000a et 2000b), qui tend à témoigner d’une certaine validité prédictive des méthodes. On ne peut cependant que rester relativement 45 - facteurs de performance (à l’aide d’une équation linéaire) est inadéquat. On peut associer à ces critiques l’idée que le modèle de fiabilité que met en avant THERP est tautologique : l'opérateur fait une erreur parce qu'il fait une erreur, et tous les opérateurs commettent les mêmes erreurs. Ceci renvoie à une ambiguïté fondamentale sur le concept même d’erreur humaine : elle recouvre à la fois la motivation (ou le diagnostic) qui conduit à une action, l'action elle-même, et ses conséquences, ce qui pose la question de ce que mesure réellement la probabilité d'erreur humaine (Hollnagel, 2000). Enfin, à supposer qu'il existe une "probabilité d'erreur humaine" absolue, son opérationnalisation pose problème, dans la mesure où les bases de données d'erreurs ne devraient en toute rigueur être utilisées que dans un contexte extrêmement restreint. THERP a connu de multiples révisions pour prendre en compte les critiques qui lui étaient adressées, notamment en essayant d’intégrer quelques éléments cognitifs ou un modèle de l’opérateur un peu plus sophistiqué, tel que par exemple le modèle « Skills-Rules-Knowledge » de Rasmussen (Rasmussen, 1986), qui précise les différents fondements des comportements suivant la tâche que l’opérateur a à accomplir. Il n’en reste pas moins que les arguments que nous avons évoqués restent largement valables. La méthode CREAM n’est elle-même pas exempte de critiques, même si elle repose justement sur un effort de dépassement des limites des méthodes mécanistes. Outre la rigueur parfois discutable de la méthode, en particulier en ce qui concerne la quantification37, elle repose sur la même objectification du suspicieux, malgré ces bons résultats, au regard des constats de Kirwan (1982) et William (1985) que rapporte Reason (1990), ou de l’étude de la Nuclear Regulatory Commission américaine que reprend Llory (1992). Ces trois études laissent en effet penser d’une part que les données finales obtenues par l’usage de méthodes d’évaluation de la fiabilité humaine ne correspondent pas nécessairement à la réalité, et d’autre part que l’application de ces méthodes par plusieurs analystes aboutit à des résultats sensiblement différents, souvent de plusieurs ordres de grandeurs. 37 Certains calculs peuvent aboutir, par le jeu des facteurs de pondération, à des probabilités supérieures à un... L’argument d’Hollnagel sur ce point est qu’il suffit alors, soit de considérer que la probabilité est égale à un, soit de renormer les résultats, ce qui est loin d’être entièrement convaincant. D’autre part, les probabilités de base sont issues pour certaines d’entre elles des mêmes méthodes que l’auteur critique dans les chapitres précédant la présentation de sa propre approche. Le fait de les présenter dans une fourchette n’est qu’un pis-aller peu convaincant. 46 - comportement de l’opérateur que les méthodes de première génération. Ainsi, il est par exemple significatif que des méthodes (tant celles de la première que celles de la deuxième génération) qui s’intéressent au comportement d’un opérateur en situation potentielle de risque ne fassent nulle part mention de l’attitude vis-à-vis du risque de ce dernier. Il existe ici un gouffre entre d’une part des techniques, qui, même avec une dose « d’ingénierie cognitive », restent d’une grande pauvreté quant à la prise en compte du comportement humain38, et d’autre part les approches des théoriciens de la décision, qui manquent d’une certaine opérationnalisation dans ce contexte, et posent à leur tour d’autres questions quant à l’intentionnalité systématique d’un comportement risqué, laquelle peut souvent apparaître comme un argument facile pour, en cas d’accident, dédouaner le management de toute responsabilité, comme l’écrit par exemple Tombs (1991). A ce titre les approches de Svenson (1998), Baumont et al. (2000), ou Pyy (2000), qui reposent explicitement sur l’analyse de la prise de décision de l’opérateur39 constituent sans doute des premiers pas intéressants, même s’ils sont vraisemblablement trop timorés. In fine, il semble bien que si les outils d’analyse des tâches, et pour partie l’analyse qualitative de la fiabilité humaine sont aujourd’hui matures et riches d’enseignements pour la compréhension des risques, l’analyse quantitative de la fiabilité humaine est en revanche cruellement en manque d’un changement radical de paradigme. 1.3. La dimension organisationnelle du management des risques industriels Pour tâcher d’aller au-delà des limites des méthodes que nous venons d’exposer, de nombreux chercheurs se sont intéressés à la prise en compte des 38 Nous examinerons dans la section suivante les approches qui enrichissent l’analyse de la fiabilité humaine par la prise en compte de l’organisation et des décisions managériales qui la conditionnent largement. Nous nous intéressons ici essentiellement aux aspects d’ordre individuel de la performance humaine. 39 On relèvera également dans cette direction les recherches du groupe de travail germano-franconéerlandais EARTH (Mosneron-Dupin et al., 1997). 47 - facteurs organisationnels et managériaux dans l’évaluation des risques (PatéCornell, 1990). Sans vouloir être aussi exhaustif qu’Abramovici (1999) ou Øien (2001), nous distinguerons ici trois principaux courants d’analyse40. Une première direction, suivie par un groupe de recherche autour du Professeur Elisabeth PatéCornell à l’Université Stanford, a consisté à étendre le modèle canonique de management des risques en modélisant les diverses relations entre facteurs managériaux et organisationnels, décisions et fonctionnement du système à l’aide de diagrammes d’influence. Une deuxième direction, développée quant à elle notamment autour des Professeurs Todd La Porte, Gene Rochlin, et Karlene Roberts à l’Université de Californie à Berkeley, s’est intéressée aux facteurs qui font que certaines organisations se caractérisent par leur haute fiabilité (High Reliability Organizations, Roberts, 1990a). Enfin, un dernier courant a analysé plus généralement l’impact de la « culture de sécurité » sur le fonctionnement des organisations en mettant l’accent sur les facteurs susceptibles de conduire aux actions erronées. Nous allons examiner successivement ces trois courants par ordre de généralité croissant. 1.3.1. L’école de Stanford et l’approche SAM « L’école de Stanford » tout d’abord, que nous avons déjà partiellement évoquée, s’est largement inspirée des travaux de Ron Howard et du Strategic Decision Group en analyse de la décision (Howard et Matheson, 1989), et plus particulièrement des diagrammes d’influence. Avant de préciser les apports de l’équipe du Professeur Paté-Cornell à la compréhension des mécanismes organisationnels sous-jacents aux défaillances techniques, il est utile ici de rappeler rapidement quelques notions d’analyse de la décision. 40 Nous laisserons de côté les approches par trop techniques de l’analyse des facteurs humains et organisationnels à l’image de celle de Bea (2002), qui constituent plus de simples extensions de THERP qu’une réelle analyse de l’organisation, et sont par voie de conséquence soumises aux mêmes critiques que celles que nous avons exposées précédemment. 48 - Un diagramme d’influence est un réseau bayésien41 enrichi par des nœuds de décision (représentés par des rectangles) et des nœuds déterministes (représentés par des rectangles aux angles arrondis)42, dont la figure 3 représente un exemple : COÛT STRATÉGIE DE DÉPLOIEMENT PROFIT SUCCÈS REVENUS Figure 4 : Exemple de diagramme d’influence Le diagramme d’influence ci-dessus traduit un problème de décision où l’on a à choisir entre plusieurs stratégies de déploiement (par exemple déploiement national, européen, international). Chacun de ces modes de déploiement a un coût différent (supposé connu avec certitude), et une probabilité de succès différente. Le succès va à son tour conditionner des revenus. Au total, revenus et coûts permettent de calculer un profit espéré. Les nœuds de décision seront spécifiés par les possibilités entre lesquelles le décideur peut choisir et les éventuelles conséquences associées à ces possibilités, les nœuds de chance par les états du monde auxquels ils peuvent conduire, par les probabilités qui les caractérisent et par les conséquences associées ; enfin, les nœuds déterministes 41 C’est-à-dire un graphe orienté acyclique dont les nœuds correspondent à des distributions de probabilités et les arcs à des conditionnements (ou dépendances) entre ces distributions. On fait parfois une distinction entre nœuds de valeur et nœuds déterministes, les premiers représentant l’objectif à atteindre (comme par exemple une valeur actuelle nette), les seconds étant de simples paramètres. En pratique, cette distinction n’est pas fondamentale, à tel point qu’elle n’est pas présente dans certains outils logiciels qui permettent d’effectuer des modélisations à base de diagrammes d’influence. Les nœuds de valeur (entendus ici au sens large) ne sont en principe pas absolument nécessaires. En pratique, ils servent cependant largement à clarifier la représentation, en isolant les conséquences associées aux nœuds de chance et de décision. Ce faisant, ils permettent aussi de mener des analyses multiattributs et simplifient également grandement les analyses de sensibilité. 49 - prennent simplement les valeurs qu’on leur donne. Les arcs quant à eux peuvent prendre plusieurs significations suivant leur origine et le nœud vers lequel ils pointent. Un conditionnement qui part d’un nœud de chance A vers un nœud de chance B traduira l’idée que les probabilités et/ou les conséquences associées aux états du monde auxquels mène la réalisation du nœud B vont dépendre de la réalisation de la variable aléatoire que représente le nœud A. Le conditionnement d’un nœud de chance B par un nœud de décision A signifiera que les probabilités et/ou les conséquences associées à B vont varier suivant le choix pris au niveau du nœud A. Inversement, le conditionnement d’un nœud de décision B par un nœud de chance A signifie que les conséquences du choix B vont dépendre de la réalisation de A : le décideur connaîtra le résultat de la réalisation de A lorsqu’il fera le choix B. Enfin, un conditionnement entre deux nœuds de décision va impliquer que le choix effectué au nœud qui est à l’origine du conditionnement va avoir un impact sur les conséquences du choix de l’autre. Les diagrammes d’influence ont un triple intérêt au moins : tout d’abord, ils permettent une représentation graphique simplifiée d’un problème complexe. En cela, ils constituent un outil particulièrement puissant de communication. Ensuite, ils résument efficacement les données d’un problème de manière synthétique pour l’analyse décisionnelle. Enfin, utilisés conjointement avec des arbres de décision dans le cadre de programmes informatiques, ils permettent de trouver facilement la solution de problèmes de décision (Shachter, 1986). Paté-Cornell et son équipe utilisent les diagrammes d’influence dans ces trois dimensions de communication, de réduction de la complexité d’un problème, et d’outil d’aide à la décision (Paté-Cornell et Regan, 1998)43. L’approche SAM (System-Action-Management) (Murphy, 1994 ; Paté-Cornell et Murphy, 1996) opère ainsi une distinction entre le système physique (associé aux outils classiques d’analyse de risques), le système des décisions et actions associées aux opérateurs, et enfin le système managérial et organisationnel, qui correspond aux 43 Embrey (1992) et Øien (2001) présentent également des analyses des facteurs organisationnels à base de diagrammes d’influence, moins riches et abouties cependant. 50 - facteurs externes qui vont influencer le système. L’analyse part du système physique, pour remonter aux facteurs organisationnels et managériaux, et utilise les règles de calculs des probabilités conditionnelles telles qu’elles s’exprime dans le diagramme d’influence pour aboutir à la quantification finale des risques. Ainsi, on peut exprimer de manière simple la probabilité de défaillance du système p(F) comme la somme des probabilités des événements IEi susceptibles de mener à la défaillance compte tenu de la probabilité p(F \| IEi) qu’ils mènent effectivement à la défaillance, qui s’exprime comme une probabilité conditionnelle. On a : p( F ) = ∑ p(F IE i ) p(IE i ) i A un second niveau, les décisions et actions des individus (qu’il s’agisse d’opérateurs, d’ingénieurs chargés du design de l’installation...) DAj vont avoir € un impact sur la probabilité de défaillance, directement et par le biais de leur influence sur la probabilité d’occurrence des événements susceptibles de mener à la défaillance. On a donc : ( p( F ) = ∑ ∑ p(F IE i ,DA j ) p(IEi DA j ) p DA j i j ) On notera ici que les décisions et actions des individus sont considérées comme des phénomènes stochastiques.	17,203
https://openalex.org/W4286907243	OpenAlex	Open Science	CC-By	2,021	Motion Control of Redundant Robots with Generalised Inequality Constraints	Amirhossein Kazemipour	English	Spoken	3,125	5,740	2021 I-RIM Conference October 8-10, Rome, Italy ISBN: 9788894580525 DOI:10.5281/zenodo.5900613 2021 I-RIM Conference October 8-10, Rome, Italy ISBN: 9788894580525 DOI:10.5281/zenodo.5900613 Motion Control of Redundant Robots with Generalised Inequality Constraints Khaled Al Khudir∗∗ Alessandro De Luca∗ Maram Khatib∗ Amirhossein Kazemipour∗ limits at the saturation level, as opposed to what happens in [10]. This is neither necessary nor optimal, and will often lead to high-frequency oscillations at the level of commands. In this paper, we propose several improvements to the SNS algorithm at the velocity level introduced in [10]. First, both joint and Cartesian inequality (box) constraints are treated as hard bounds. Second, all inequalities are assigned the same priority, i.e., enforced anyway independently of the end- effector (EE) task (or simply considered out of the SoT, in case of multiple tasks). Finally, the modiﬁcations are made so as to preserve the automatic optimal task scaling of the original SNS approach, which relaxes the primary task (keeping its ge- ometric direction) only when no feasible solution would exist, while guaranteeing satisfaction of all inequality constraints. The resulting algorithm does not need parameter tuning and is faster than QP solvers. limits at the saturation level, as opposed to what happens in [10]. This is neither necessary nor optimal, and will often lead to high-frequency oscillations at the level of commands. In this paper, we propose several improvements to the SNS algorithm at the velocity level introduced in [10]. First, both joint and Cartesian inequality (box) constraints are treated as hard bounds. Second, all inequalities are assigned the same priority, i.e., enforced anyway independently of the end- effector (EE) task (or simply considered out of the SoT, in case of multiple tasks). Finally, the modiﬁcations are made so as to preserve the automatic optimal task scaling of the original SNS approach, which relaxes the primary task (keeping its ge- ometric direction) only when no feasible solution would exist, while guaranteeing satisfaction of all inequality constraints. The resulting algorithm does not need parameter tuning and is faster than QP solvers. Abstract—We present an improved version of the Saturation in the Null Space (SNS) algorithm for redundancy resolution at the velocity level. In addition to hard bounds on joint space motion, we consider also Cartesian box constraints that cannot be violated at any time. The modiﬁed algorithm combines all bounds into a single augmented generalised vector and gives equal, highest priority to all inequality constraints. When needed, feasibility of the original task is enforced by the SNS task scaling procedure. Simulation results are reported for a 6R planar robot. I. INTRODUCTION Given a m-dimensional primary task to be performed by a robot with n joints, with n > m (redundancy), a standard method to prevent violation of joint/Cartesian inequalities during motion is to resort to some form of artiﬁcial poten- tials [1], pushing away from their limits the joints and the control points on the robot body [2]. However, this method is highly parameter-dependent and may introduce oscillations when activating/deactivating the avoidance task in proximity of the bounds [3]. To milden such undesired behavior, the null-space projection term or the activation function can be designed in an incremental way [4], [5]. Nonetheless, selection of appropriate gains is still needed. Moreover, in case of multiple tasks, incorporating the avoidance behavior in the original Stack of Tasks (SoT) will assign different priorities to each single constraint [4]–[7]. Motion Control of Redundant Robots with Generalised Inequality Constraints Index Terms—Motion control, Redundant robots, Inequality constraints, Hard limits. ∗Dipartimento di Ingegneria Informatica, Automatica e Gestionale, Sapienza Universit`a di Roma, Via Ariosto 25, 00185 Roma, Italy. Emails: amrkzp@gmail.com, khatib@diag.uniroma1.it, deluca@diag.uniroma1.it. ∗∗School of Mechanical, Aerospace and Automotive Engineering, Coventry University, CV1 5FB Coventry, UK. Email: khaled.alkhudir@coventry.ac.uk. A. Generalised constraints Consider a robot with n joints and r generic Cartesian control points distributed on the robot body, each of dimension di ∈{1, 2, 3}, i = 1, . . . , r. We deﬁne an augmented vector a = qT pT cp,1 pT cp,2 . . . pT cp,r T , (1) (1) where q ∈Rn denotes the joint variables and pcp,i ∈Rdi is the position of the i-th control point, i = 1, . . . , r. The joint variables as well as the Cartesian control points will have some desired motion restrictions. Accordingly, we deﬁne the augmented matrix Other numerical approaches incorporate joint space and Cartesian motion limits as inequality constraints using parameter-free optimization, such as Quadratic Programming (QP) [8], [9]. However, these methods are computationally slower than analytical solutions and do not lead to realizable solutions when the original task(s) is not compatible with the set of inequality constraints. The SNS algorithm introduced in [10] links QP to the SoT approach and overcomes these challenges. In the original paper, joint motion limits were considered as hard bounds (i.e., that cannot be relaxed in a least-square sense) and treated out of the SoT. On the other hand, Cartesian (avoidance) constraints were not handled as hard bounds. In [11], an approach has been proposed to include joint and Cartesian inequality constraints in the SoT for torque-controlled manipulators. However, joint limits are always pre-assigned the highest priority over all other constraints. Moreover, all violated constraints are set to their A = I JT cp,1 JT cp,2 . . . JT cp,r T , (2) (2) where I ∈Rn×n is the identity matrix and Jcp,i ∈Rdi×n is the Jacobian matrix of the i-th control point. Deﬁne the position and velocity limits for each joint, j = 1, . . . , n, as Qmin j ≤qj ≤Qmax j , V min j ≤˙qj ≤V max j , (3) (3) and the limits for each control point, i = 1, . . . , r, P min cp,i ≤pcp,i ≤P max cp,i , V min cp,i ≤˙pcp,i ≤V max cp,i . ∗∗School of Mechanical, Aerospace and Automotive Engineering, Coventry University, CV1 5FB Coventry, UK. Email: khaled.alkhudir@coventry.ac.uk. ∗Dipartimento di Ingegneria Informatica, Automatica e Gestionale, Sapienza Universit`a di Roma, Via Ariosto 25, 00185 Roma, Italy. Emails: amrkzp@gmail.com, khatib@diag.uniroma1.it, deluca@diag.uniroma1.it. B. Modiﬁed SNS algorithm (11) Consider a single EE velocity task ˙xd ∈Rm, with n > m, and its Jacobian matrix J ∈Rm×n, to be achieved under the generalised hard constraints in (7). The pseudo-code of the modiﬁed SNS method is presented in Algorithm 1. If the Cartesian inequality limits in (4) are discarded, the augmented matrix (2) becomes A = I and it is easy to show that Algorithm 1 simpliﬁes to the original SNS algorithm in [10]. Note that at line 15, the most critical constraint corresponds to the smallest scaling factor sk over all constraints. Also, at line 19, when there is no way to perform the desired task under the hard inequality constrains, we apply an optimal task scaling factor as computed by Algorithm 2, similar to [10]. The EE starts the motion very close to the desired path. As shown in Fig. 2, the positional error converges immediately and remains zero along the whole task. Accordingly, the task scaling is active (s < 1) only for few milliseconds at beginning, to comply with the saturated joint and Cartesian velocity limits due to the initial error recovery —see Figs. 3 and 4. Later, the robot is able to perform the complete task perfectly while satisfying all inequality constraints (many of them in saturation). The few discontinuities in the commanded joint velocity in Fig. 3 can be addressed by extending the algorithm to the acceleration level and including suitable joint acceleration limits in the set of constraints. The EE starts the motion very close to the desired path. As shown in Fig. 2, the positional error converges immediately and remains zero along the whole task. Accordingly, the task scaling is active (s < 1) only for few milliseconds at beginning, to comply with the saturated joint and Cartesian velocity limits due to the initial error recovery —see Figs. 3 and 4. Later, the robot is able to perform the complete task perfectly while satisfying all inequality constraints (many of them in saturation). The few discontinuities in the commanded joint velocity in Fig. 3 can be addressed by extending the algorithm to the acceleration level and including suitable joint acceleration limits in the set of constraints. A. Generalised constraints The sampling time is T = 1 [ms] and the initial robot conﬁguration (in [rad]) is chosen as with the control gain matrix Kp = diag{50, 50} and the EE position xee computed by the direct kinematics. The sampling time is T = 1 [ms] and the initial robot conﬁguration (in [rad]) is chosen as and ˙P min cp,i = max ( P min cp,i −pcp,i T , V min cp,i ) , ˙P max cp,i = min P max cp,i −pcp,i T , V max cp,i , (6) q0 = π 6 −π 6 −π 6 π 3 −π 6 −π 6 T . (9) (9) The limits (3) are equal and symmetric for all joints: where T is the sampling time. Accordingly, the generalised inequality constraints can be written as the augmentation of joint and Cartesian bounds in (5) and (6) as where T is the sampling time. Accordingly, the generalised inequality constraints can be written as the augmentation of joint and Cartesian bounds in (5) and (6) as Qmax j = −Qmin j = π 2 [rad], V max j = −V min j = 1 [rad/s]. (10) (10) Bmin = ˙Qmin,1, . . . ˙Qmin,n, ˙P min cp,1 , . . . ˙P min cp,r T , Bmax = ˙Qmax,1, . . . ˙Qmax,n, ˙P max cp,1 , . . . ˙P max cp,r T . (7) Bmin = ˙Qmin,1, . . . ˙Qmin,n, ˙P min cp,1 , . . . ˙P min cp,r T , Bmax = ˙Qmax,1, . . . ˙Qmax,n, ˙P max cp,1 , . . . ˙P max cp,r T . (7) ( ) We consider r = 5 control points (each with di = 1) along the robot body, located at the joints j = 2, . . . , 6. The corresponding limits (4) are equal for all points, and imposed only over the y-direction: P max,y cp,i = 1 [m], P min,y cp,i = −1.1 [m], V max,y cp,i = −V min,y cp,i = 0.8 [m/s]. (11) A. Generalised constraints (4) At a generic time instant, the box constraints for the velocity of each component of (1) are given by ˙Qmin,j = max ( Qmin j −qj T , V min j ) , ˙Qmax,j = min Qmax j −qj T , V max j , (5) (5) 138 Algorithm 1 SNS with generalised inequality constraints ˙qN ←0, s∗←0, P ←I, Alim ←null, ˙aN ←null 2: repeat limits violated ←FALSE 4: ˙q ←˙qN + (J P )# ( ˙x −J ˙qN) ˙a ←A ˙q 6: if ∃h ∈ 1 : n + Σr 1di : (˙ah < bmin,h) ∨(˙ah > bmax,h) then limits violated ←TRUE 8: α ←A (J P )# ˙x β ←˙a −α 10: getTaskScalingFactor(α, β) if {task scaling factor} > s∗then 12: s∗←{task scaling factor} ˙q∗ N ←˙qN, P ∗←P 14: end if k ←{the most critical constraint} 16: Alim ←concatenate(Alim, Ak) ˙aN ← concatenate(˙aN, bmax,k) if (˙ah > bmax,k) concatenate(˙aN, bmin,k) if (˙ah < bmin,k) 18: P ←I −(Alim)# (Alim) if rank(JP ) < m then 20: ˙q ←˙q∗ N + (J P ∗)# s∗˙x −J ˙q∗ N limits violated ←FALSE 22: end if end if 24: ˙qN ←(Alim)# ˙aN until limits violated = TRUE 26: ˙qSNS ←˙q Algorithm 2 Optimal task scaling factor function GETTASKSCALINGFACTOR(α, β) 2: for h ←1 : n + Σr 1di do Lh ←bmin,h −βh 4: Uh ←bmax,h −βh if αh < 0 ∧Li < 0 then 6: if αh < Lh then sh ←Lh/αh 8: else sh ←1 10: end if else if αh > 0 ∧Uh > 0 then 12: if αh > Uh then sh ←Uh/αh 14: else sh ←1 16: end if else 18: sh ←0 end if 20: end for return s 22: end function EE is required to track a 2D linear path (m = 2), see Fig. 1. Therefore, the primary EE velocity task is deﬁned as Algorithm 1 SNS with generalised inequality constraints Algorithm 2 Optimal task scaling factor EE is required to track a 2D linear path (m = 2), see Fig. 1. Therefore, the primary EE velocity task is deﬁned as ˙x = ˙xd + Kp(xd −xee), (8) (8) with the control gain matrix Kp = diag{50, 50} and the EE position xee computed by the direct kinematics. pp [11] J. D. M. Osorio, F. Allmendinger, M. D. Fiore, U. E. Zimmermann, and T. Ortmaier, “Physical human-robot interaction under joint and cartesian constraints,” in Proc. 19th Int. Conf. on Advanced Robotics, pp. 185– 191, 2019. IV. CONCLUSION Veriﬁcation for the proposed algorithm is done through MATLAB simulations, using a 6R planar robot arm (n = 6) and considering joint and Cartesian inequality constraints. The We have proposed major enhancements to the basic SNS algorithm at the velocity level for redundant robots. Cartesian 139 0 2 4 6 8 10 -1 0 1 0 2 4 6 8 10 -1 0 1 Fig. 3: Evolution of the joints during task execution. The dotted red lines are the bounds on the joint motion. 0 2 4 6 8 10 -1 0 1 0 2 4 6 8 10 -1 0 1 0 1 2 3 4 5 6 -1.5 -1 -0.5 0 0.5 1 1.5 Fig. 1: Initial (black) and ﬁnal (gray) conﬁgurations of the 6R planar arm. The red circles represent the robot joints (and the EE tip). The desired EE path is the blue line, to be traced from right to left. The dashed red lines are the Cartesian position limits. The dashed green lines show the path of control points during task execution. 0 1 2 3 4 5 6 -1.5 -1 -0.5 0 0.5 1 1.5 Fig. 1: Initial (black) and ﬁnal (gray) conﬁgurations of the 6R planar arm. The red circles represent the robot joints (and the EE tip). The desired EE path is the blue line, to be traced from right to left. The dashed red lines are the Cartesian position limits. The dashed green lines show the path of control points during task execution. Fig. 3: Evolution of the joints during task execution. The dotted red lines are the bounds on the joint motion. 0 2 4 6 8 10 -0.1 0 0.1 0 2 4 6 8 10 0 0.5 1 0 2 4 6 8 10 -0.1 0 0.1 0 2 4 6 8 10 0 0.5 1 Fig. 2: EE positional errors and related task scaling factor. 0 1 2 3 4 5 6 7 8 9 10 -1 -0.5 0 0.5 1 0 1 2 3 4 5 6 7 8 9 10 -1 -0.5 0 0.5 1 Fig. 4: Evolution of the control points along the y-direction. The dotted red lines are the Cartesian bounds on the motion of the control points. pp [10] F. Flacco, A. De Luca, and O. Khatib, “Control of redundant robots under hard joint constraints: Saturation in the null space,” IEEE Trans. on Robotics, vol. 31, no. 3, pp. 637–654, 2015. [8] A. Escande, N. Mansard, and P.-B. Wieber, “Hierarchical quadratic programming: Fast online humanoid-robot motion generation,” Int. J. of Robotics Research, vol. 33, no. 7, pp. 1006–1028, 2014. IV. CONCLUSION 0 1 2 3 4 5 6 7 8 9 10 -1 -0.5 0 0.5 1 0 1 2 3 4 5 6 7 8 9 10 -1 -0.5 0 0.5 1 Fig. 2: EE positional errors and related task scaling factor. inequality constraints are included and treated as hard limits, while preserving all the nice features of the original method. The modiﬁed algorithm can be extended to include multiple tasks having different priorities. Moreover, it can be imple- mented also at the acceleration level, which is beneﬁcial for involving dynamic properties in the resolution of redundancy and is suitable for torque-controlled systems. pp [9] E. M. Hoffman, A. Laurenzi, L. Muratore, N. G. Tsagarakis, and D. G. Caldwell, “Multi-priority Cartesian impedance control based on quadratic programming optimization,” in Proc. IEEE Int. Conf. on Robotics and Automation, pp. 309–315, 2018. [6] L. Sentis and O. Khatib, “Synthesis of whole-body behaviors through hierarchical control of behavioral primitives,” Int. J. of Humanoid Robotics, vol. 2, no. 4, pp. 505–518, 2005. [8] A. Escande, N. Mansard, and P.-B. Wieber, “Hierarchical quadratic programming: Fast online humanoid-robot motion generation,” Int. J. of Robotics Research, vol. 33, no. 7, pp. 1006–1028, 2014. [9] E. M. Hoffman, A. Laurenzi, L. Muratore, N. G. Tsagarakis, and D. G. Caldwell, “Multi-priority Cartesian impedance control based on quadratic programming optimization,” in Proc. IEEE Int. Conf. on Robotics and Automation, pp. 309–315, 2018. [10] F. Flacco, A. De Luca, and O. Khatib, “Control of redundant robots under hard joint constraints: Saturation in the null space,” IEEE Trans. on Robotics, vol. 31, no. 3, pp. 637–654, 2015. [11] J. D. M. Osorio, F. Allmendinger, M. D. Fiore, U. E. Zimmermann, and T. Ortmaier, “Physical human-robot interaction under joint and cartesian constraints,” in Proc. 19th Int. Conf. on Advanced Robotics, pp. 185– 191, 2019. REFERENCES Fig. 4: Evolution of the control points along the y-direction. The dotted red lines are the Cartesian bounds on the motion of the control points. [1] O. Khatib, “Real-time obstacle avoidance for manipulators and mobile robots,” in Autonomous Robot Vehicles (I. Cox and G. Wilfong, eds.), pp. 396–404, Springer, 1986. pp p g [2] M. Khatib, K. Al Khudir, and A. De Luca, “Task priority matrix at the acceleration level: Collision avoidance under relaxed constraints,” IEEE Robotics and Automation Lett., vol. 5, no. 3, pp. 4970–4977, 2020. [3] M. Khatib, K. Al Khudir, and A. De Luca, “Task priority matrix under hard joint constraints,” in Proc. 2nd Italian Conf. on Robotics and Intelligent Machines, pp. 173–174, 2020. [4] N. Mansard, O. Khatib, and A. Kheddar, “A uniﬁed approach to integrate unilateral constraints in the stack of tasks,” IEEE Trans. on Robotics, vol. 25, no. 3, pp. 670–685, 2009. [5] E. Simetti and G. Casalino, “A novel practical technique to integrate inequality control objectives and task transitions in priority based control,” J. of Intelligent & Robotic Systems, vol. 84, no. 1, pp. 877–902, 2016. [6] L. Sentis and O. Khatib, “Synthesis of whole-body behaviors through hierarchical control of behavioral primitives,” Int. J. of Humanoid Robotics, vol. 2, no. 4, pp. 505–518, 2005. pp [7] L. Sentis and O. Khatib, “A whole-body control framework for hu- manoids operating in human environments,” in Proc. IEEE Int. Conf. on Robotics and Automation, pp. 2641–2648, 2006. 140	6,272
https://openalex.org/W4300278976	OpenAlex	Open Science	Public Domain	1,989	An improved operating mode for a Si(Li) x-ray spectrometer	N W Madden	English	Spoken	3,915	8,009	LBL--27067 DE91 000358 LBL--27067 DE91 000358 LBL--27067 DE91 000358 Abstract _- ...._ ....... s,,.,,'_ '1"4" ---N t.v) % "_ \"\._.. ..... _,_ "N_" ' ' _" _'"_ =td' L .....b "'" b.... _t-o r4,m eS) Fig. 1: Plot of FWHM resoluUon squared vs. peaking time of Si(Li) X-ray spectrometer displaying the series, parallel and 1/f noise components. A new technique has been developed for restoring the charge to the feedback capacitor in charge sensitive pream- plifiers. The method, termed "transient-reset," was devel- oped for use in very low noise Si(Li) X-ray spectrometers. lt has proved valuable as a diagnostic tool for evaluating the noise components of the input circuit in these systems. The technique is described and compared with the pulsed- light and transistor-reset methods. To complement the charge restorationtechnique, a new configuration of Si(Li) X-raydetecter witha verythinaluminumentrance window has been developed. Measurements of the effective window thickness are compared with similar results obtained on gold nickel and ion-implanted boron contacts AN IMPROVED OPERATING MODE FOR A Si(Li) X-RAY SPECTROMETER N. W. Madden, F. S. Goulding, J. M. Jaklevic, D. A. Landis and C. S. Rossington Engineering Division Lawrence Berkeley Laboratory 1 Cyclotron Rd. Berkeley, CA 94720 N. W. Madden, F. S. Goulding, J. M. Jaklevic, D. A. Landis and C. S. Rossington Engineering Division Lawrence Berkeley Laboratory 1 Cyclotron Rd. Berkeley, CA 94720 I. SCIENTWIC MOTIVATION In Fig 2B a smaller device by Interfet exhibits higher spectrometer measured with a 6th order pseudo-Gaussian pulse shaper. The flat minimum in the noise plot shows the dominance of 1/f noise which limits the resolution of the system for X-ray energies less than 1keV. Series 1/f noise is associated, at low operating temperatures, with trapping/detrapping at impurities and defects in the con- ducting channel of the FET. Parallel 1/f noise, on the other hand, is believed to originate in lossy dielectrics which are capacitively coupled into the FET gate, and possibly in sur- face layers of the detectors [1]. Careful control of impuri- ties in the FET manufacturing process has reduced the mag- nitude of the series 1/f noise. In some Interfet devices, the llf noise is less than the channel series noise at ali frequen- ties down to 3 Hz [2]. One way to determine the presence of trapping/detmpping noise at sites in the FET channel is to performnoisemeasurements at different temperaturesand fr_uencies in a grounded:gate configuration. Parallel input circuit noise is eliminated in this case. Figures 2A and 2B show such measurements of two small geometry N-channel junction FrETs[3]. In Fig. 2A, a Motorola device clearly exhibits bumps in th, plot that accompany series 1/f noise. In Fig 2B a smaller device by Interfet exhibits higher AN IMPROVED OPERATING MODE FOR A Si(Li) X-RAY SPECTROMETER N. W. Madden, F. SI Goulding, J. M. Jaklevic, D. A. Landis and C,, S. Rossington Engineering Division Lawrence Berkeley Laboratory University of California 1 Cyclotron Rd. Berkeley, CA 94720 This work was supported by the Director, Office of Energy Research, Office of He_/dthand Environmental Research, Physical and Technological Division of the U.S. Department of Energy under Contract No. DE-AC03-76SFIX_8. This work was supported by the Director, Office of Energy Research, Office of He_/dthand Environmental Research, Physical and Technological Division of the U.S. Department of Energy under Contract No. DE-AC03-76SFIX_8. This work was supported by the Director, Office of Energy Research, Office of He_/dthand Environmental Research, Physical and Technological Division of the U.S. Department of Energy under Contract No. DE-AC03-76SFIX_8. LBL-27067 I. SCIENTWIC MOTIVATION Fig. 1: Plot of FWHM resoluUon squared vs. peaking time of Si(Li) X-ray spectrometer displaying the series parallel and 1/f noise components The problem of "dead" layers at ',.heentry windo6, of Si(Li) X-ray detectors is common to zli low-energy X-ray applications of these detectors. One such application is to grazing angle X-ray telescopes that have made possible ob- servations of weak stellar X-ray sources. The focal plane detectors used in many of these tele_opes are gas propor- tional flow counters and Si(Li) X-ray detectors. Gas pro- '7,ortionalcounters can be made with large areas but have only moderate energy resolution at low energies. They are used frequently with short focal length telescopes. Newer telescopes, such as that designed for the broad band X-ray telescope "BBXRT" project at Goddard Space Flight Cen- ter, have small focal spots (4-5mm) which match Si(Li) X- ray detectorsthat offer excellent energy resolution. Since these instruments operme in the vacuum of _ace it is pos- sible to observe low-energy X-rays such as those,from car. bon, boron and beryllium if the spectrometer has suitably low noise and very thin entrance windows. The work de- scribed hea_was aimed at these applications, spectrometer measured with a 6th order pseudo-Gaussian pulse shaper. The flat minimum in the noise plot shows the dominance of 1/f noise which limits the resolution of the system for X-ray energies less than 1keV. Series 1/f noise is associated, at low operating temperatures, with trapping/detrapping at impurities and defects in the con- ducting channel of the FET. Parallel 1/f noise, on the other hand, is believed to originate in lossy dielectrics which are capacitively coupled into the FET gate, and possibly in sur- face layers of the detectors [1]. Careful control of impuri- ties in the FET manufacturing process has reduced the mag- nitude of the series 1/f noise. In some Interfet devices, the llf noise is less than the channel series noise at ali frequen- ties down to 3 Hz [2]. One way to determine the presence of trapping/detmpping noise at sites in the FET channel is to performnoisemeasurements at different temperaturesand fr_uencies in a grounded:gate configuration. Parallel input circuit noise is eliminated in this case. Figures 2A and 2B show such measurements of two small geometry N-channel junction FrETs[3]. In Fig. 2A, a Motorola device clearly exhibits bumps in th, plot that accompany series 1/f noise. DI. CHARGE RESTORATION Virtually ali modem low noise preamplifiers used in conjunction with Si(Li) X-ray spectrometers employ a charge sensitive stage. Charge restoration is accomplished by either pulsed-light reset [4] or one of the several variants of transistor-reset [5], Figure 3A is a schematic representa- tion of a pulsed-light reset preamplifier. The detector cur- rent flows from the virtual ground (gate of the bET) to the negative detector bias. If the preamplifier is operatedwith- out detector current flowing, it will immediately saturate, due to the lET drain-gate leakage, precluding any mea- smements of the lET. In normal operation, when the bET is operated with a reverse-bi_ed detector, the companion LED will be turned on when the preamplifier output reaches a predetermined level. A large positive current (nA-l.tA) then flows into the virtual ground, the result of illuminat- ing the FETs drain-to-gate diode. As the charge is restored and the preamplifier output reaches a predetermined level, the LED is switched off and the cycle begins again. Pulsed,light reset was originally designed to minimize in- put circuit components. However, drawbacks include the inability to handle reverse-polarity detectors, the require- ment of a light-tight package for the FET and LED, and the impairment of resolution at high rates due to residual after- effects of the light [4]. p ( ) 5 2 '_\! _ ' I < "r'N10t \ lo (Hz) > ,.- t-. w 5 30 (Hz) .a.- ;_5 (Hz) ,_-sm (Hz) tK, 3K, 1_( L i_d,I 1_ (Hz) "'_13_ I i , , I \| I, \| i I i i t \| i i \| I i \| i i I \| _ i i ! i t i i t l_ t50 :_r_ 250 sm 35e Temper atu re (K* ) Fig. 2: Plot of channel noise as a function of temperature d f f A) M l 2N4416 bET d A transistor-reset preamplifier is shown in Fig. 3B. lt usesa selected low capacitance grounded-base bipolar tran- sister to feed the reset charge to the bET gate. Either polar- ity of charge can be restored by selection of the appropriate bipolar transistor. A light-tight package is not required and very high count rates with little or no resolution degrada- tion can be achieved [6]. However, the small additional ca- pacitance (0.3-0.5 pF) introduced in the gate circuit causes a _fignificant degradation of resolution in low noise (low de- teeter capacitance) spectrometers. II. NOISE LIMITATIONS Careful analysis of the noise characteristics of very good Si(Li) X-ray spectrometers shows that the limiting noise source is 1/f noise. For example, Fig. 1 is a noise plot of a 5 2 _ ' _' <' lO _ 1_ "a- > _ t-- I w 5 'tHZ) 75 (Hz) sm (HZ) IK (Hz) 3K (Hz) (Hz) 2 1o_((Hz) _" ( Ft ) 11_ 18_ 150 208 258 388 358 Tr:mperature (K) 5 2 '_\! _ ' I < "r'N10t \ lo (Hz) > ,.- t-. w 5 30 (Hz) .a.- ;_5 (Hz) ,_-sm (Hz) tK, 3K, 1_( L i_d,I 1_ (Hz) "'_13_ I i , , I \| I, \| i I i i t \| i i \| I i \| i i I \| _ i i ! i t i i t l_ t50 :_r_ 250 sm 35e Temper atu re (K ) Fig. 2: Plot of channel noise as a function of temperature and frequency for A) Motorola 2N4416 bET and B) I t f NJ14L01 noise at the higher frequencies as expected due to the lower transconductance, but shows bumps only at very low fre- quencies indicating very little series 1/f noise. With regard to parallel 1/f noise, experience suggests that the high- purity boron nitride used to package the FET chip and its companion LED in a light-tight package is a major source of parallel 1/f noise. The new charge restoration technique avoids the use of a ptabed-light reset, and since the FET package need not be light-fight, dielectrics with much better loss characteristics than boron nitride can be used. The utilization of both lower loss dielectrics associated with the new charge restoration technique and modem selected FETs make it possible, to large},y eliminate both parallel and series 1/f noise. 5 2 _ ' _' <' lO _ 1_ "a- > _ t-- I w 5 'tHZ) 75 (Hz) sm (HZ) IK (Hz) 3K (Hz) (Hz) 2 1o_((Hz) _" ( Ft ) 11_ 18_ 150 208 258 388 358 Tr:mperature (K) DI. CHARGE RESTORATION This additional noise could be reduced by using a very low capacitance transistor i llf b i t d f thi li ti Temper atu re (K ) p ( ) Fig. 2: Plot of channel noise as a function of temperature and frequency for A) Motorola 2N4416 bET and B) Into-fea NJ14L01. Figure 3C is a schematic representation of a transient- reset preamplifier. Charge restoration is initiated when the pre,amplifier output reaches a predetermined lower level. Convent i on_l Pulsed Li ght Reset [ _5._ _--- " \| [_c.o_ ,A t o_C. t..,_l ' t_' .., '\, _ I _c@_ff_[;1 _,T _ T-"_-TL-_ _--_ l , I ,t, ,4,It_,.,,:,,. I 1! I _,.._,,_,¢,_- in _"g* - b ..... - _+,_,v,_t,,gor,_.,°,- I r.,,_°,.,,.,F,,.,,,,_, A "r,-,,.r,s'++t_,',.--me_,+'r, G:3nvent Ional SI (LI) X-ray ,_ectrometer __ [ _" _ ]_ r'..+..,++,..,.,,. iii: I, , "+ "__ m,.+ , """ ' _ TP_als_or "=- lm,,", t, _--_.' i __ J t H,_ v.,_q,r, i_.,. [ r.,,t.,,,,t P.-m I1 Tr ass test Reset rh tn _11ndow X-Ray Spectrometer "'1 Cml II, oi" i _ / DItolr'c, loin Ill The reset logic module develops a command signal which is adjustable both in amplitude and duration, to pull the base of the folded cascode transistor low. The drain of the FrETis driven just sufficiently negative with respect to the gate.to cause conduction in the gate-drain diode which re.. stores the charge on the feedback capacitor. The magnitude of the forward bias and the duration of the waveform deter- mine the quantity of charge restored. Charge is irtjected as the gate-drain diode is driven into conduction by the nega: tive going leading edge of the charge restoration waveform. Most of this charge is extracted on the positive going back edge of this waveform as the gate-drain diode returns to its normal reverse biased condition. A large voltage transient, lasting several tens of microseconds, is observed at the preamplifier output even though the width of the drain waveform is only a few microse.conds. This is a serious limitation in applying _e technique to high rate applica- tions. A detailed schematic of a transient-reset preamplifier with control logic is shown in Fig. 4. A somewhat similar charge restoration method was described in 1970 by Radeka [7]. Here charge restoration was accomplished by applying a high amplitude fast pulse train ttu'oughthe detector capac- itance, to cause the gate-to-source diode to conduct. DI. CHARGE RESTORATION c_°...,_-.m ¢ Tr ass test Reset rh tn _11ndow X R S t t DI. CHARGE RESTORATION Radeka's method minimizes dead time due to transients after charge restoration. Convent i on_l Pulsed Li ght Reset [ _5._ _--- " \| [_c.o_ ,A t o_C. t..,_l ' t_' .., '\, _ I _c@_ff_[;1 _,T _ T-"_-TL-_ _--_ l , I ,t, ,4,It_,.,,:,,. I 1! I _,.._,,_,¢,_- in _"g* - b ..... - _+,_,v,_t,,gor,_.,°,- I r.,,_°,.,,.,F,,.,,,,_, A Convent i on_l Pulsed Li ght Reset \| [_c.o_ ,A t o_C. t..,_l ' t_' .., '\, _ I _c@_ff_[;1 _,T _ T-"_-TL-_ _--_ l , I ,t, ,4,It_,.,,:,,. I 1! I _,.._,,_,¢,_- in _"g* - b ..... - _+,_,v,_t,,gor,_.,°,- I r.,,_°,.,,.,F,,.,,,,_, A "r,-,,.r,s'++t_,',.--me_,+'r, G:3nvent Ional SI (LI) X-ray ,_ectrometer __ [ _" _ ]_ r'..+..,++,..,.,,., iii: I, , "+ "__ m,.+ , """ ' _ TP_als_or "=- lm,,", t, _--_.' i __ J t H,_ v.,_q,r, i_.,. [ r.,,t.,,,,t P.-m I1 Tr ass test Reset rh tn _11ndow X-Ray Spectrometer "'1 Cml II, oi" i _ / DItolr'c, loin Ill t_,_=,:_., I _ icl c-";_.+.,+. _ i.,,,- x,, I [ [ H,_ v._,._,, r, ,_.,. c_°...,_-.m ¢ Fig. 3: Simplified schematic of three types of pulsed reset preamplifiers: A) pulse light, B) transistor reset, C) transient reset. A "r,-,,.r,s'++t_,',.--me_,+'r, G:3nvent Ional SI (LI) X-ray ,_ectrometer __ [ _" _ ]_ r'..+..,++,..,.,, iii: I, , "+ "__ m,.+ , """ ' _ TP_als_or "=- lm,,", t, _--_.' i __ J t H,_ v.,_q,r, i_.,. [ r.,,t.,,,,t P.-m I1 "r,-,,.r,s'++t_,',.--me_,+'r, G:3nvent Ional SI (LI) X-ray ,_ectrometer The new method is limited to removing positive charge from the gate circuit, and since conventional Si(Li) X-ray detectors also remove positive charge, it could not be used with such detectors. Justification for the new method lies in the fact that the new technique provides us with the means to measure the noise characteristics of an FrETwith- out thenecessity of, and therefore the ambiguity introduced by, an operating detector connected to the gate of the FRET To utilize the technique in a practical spectrometer a new detector configuration is needed that permits the gate of the lET to connect to the F'+ contact of a detector while the N+ contact connects to a positive detector bias supply I1 Tr ass test Reset rh tn _11ndow X-Ray Spectrometer "'1 Cml II, oi" i _ / DItolr'c, loin Ill t_,_=,:_., I _ icl c-";_.+.,+. _ i.,,,- x,, I [ [ H,_ v._,._,, r, ,_.,. TABLE I Fig. 5: Cross sectional view of conventional and reconfig- d l d Si(Li) X d barrier, on the other side of the detector, an evaporated alu- minum contact is made, replacing the contact that was pre- viously gold. The aluminum contact is an N+ barrier to which positive bias is applied. This contact also serves as the entrance window for X.rays. The gold contact, a P+ barrier, is connected to the gate of the input FET. Experi- ence shows that the finished detector is best tested at liquid nitrogen temperatures. barrier, on the other side of the detector, an evaporated alu- minum contact is made, replacing the contact that was pre- viously gold. The aluminum contact is an N+ barrier to which positive bias is applied. This contact also serves as the entrance window for X.rays. The gold contact, a P+ barrier, is connected to the gate of the input FET. Experi- ence shows that the finished detector is best tested at liquid nitrogen temperatures. ( ) silicon interface of conventional detectors. This canexplain the smaller dead layer ( ) silicon interface of conventional detectors. This canexplain the smaller dead layer 2) Background We have also observed a much reduced background at energies below the full energy peak for X-rays such as the Mn X-rays produced by 55Fe as seen in Fig. 6. In part this may be explained by the thinner silicon dead layer de- scribed in the previous section. An additional reason that might be,more important is illustrated by the field patterns of Figs. 7 and 8. The n-type surface channels nearly always present on silicon distort the field lines a_ shown in these figures. Comparing the conventional detector with a new reconfigured-electrode detector we see that the charge pro- duced near the window of a conventional detector may be collected in surface channels (resulting in partial signals) while that is not the case for the new configuration. This finding is further bolstered by the observation of a 0.10 Fano factor, which we have previously observed only in large volume detectors Two important observations have been made using these detectors: 1) EntranceWindow Dead Layer Using the method discussed in Refs. 8-10, a determina- t.ionhas been made of the thickness of the "dead layer", for the N. window on the _econfigured electrode detector, ex- isting within the silicon itself. Similar determinations have been made forconventional detectors using gold, nick- el and implanted boron contacts [10]. The aluminum con- tact, as seen in Table l, exhibits a much thintierdead layer than any of the other contacts. One explanation of this relies on the mechanism as discussed in Ref. 9. Charge produced by X-ray interactions in the region near the metal- silicon interface is subject to diffusion as well as drift in the electric field. Consequently, some fraction of the carrierswill reach the metal-silicon interface and may be lost or reflected depending on the recombination velocity at the interface, lt appears likely that the "reflection coeffi- cient" for holes,reaching the aluminum-silicon interface is much higher than that for electrons reaching the gold- IV. RECONFIGURED-ELECTRODE DETECTOR design The of the detector is constrained by the forego- ing considerations and by the need to achieve a thick deple- tion layer to reduce capacitance. At the present time, high- purity silicon withadequate trapping performance for broad, energy-range spectroscopy is not available, so we are.forced to use lithium-drifted material. The detector configuration employed here is shown in Fig. 5B. A conventional 3 mm thick detector, as shown in Fig. 5A, is made by normal procedures. The device is tested with particular emphasis on checking for good lithium compensation by measuring the capacitance vs. bias-voltage relationship. The lithium contact and the gold surface-barrier are then removed by lap- ping. After the detector is cleaned and etched, the original lithium contact is replaced by an evaporated gold surface- ¢ Fig. 3: Simplified schematic of three types of pulsed reset preamplifiers: A) pulse light, B) transistor reset, C) transient reset. 3 ....... r1 A ....... _+ . , Fig. 6: 55Fe spectrum measured with re,configured elec- trode detectorandtransient reset preamplLfier ....... r1 A ....... _+ . , Fig 6: 55Fe spectrum measured with reconfigured elec C_nvent ion a I Ru Bar r I• r Ih\\\,k\\\ Sl(Li) Detector I Barrier E_l] ect rode ' Ru Barrier J (N+) (p+)l , Co_en= area Reg Ion ' (,B) Fig. 5: Cross sectional view of conventional and reconfig- ttred electrode Si(Li) X-ray detectors ....... _+ . , Fig. 6: 55Fe spectrum measured with re,configured elec- trode detectorandtransient reset preamplLfier ....... _+ . , Fig. 6: 55Fe spectrum measured with re,configured elec- trode detectorandtransient reset preamplLfier TABLE I Thickness of the dead layer in the Si(Li), detector for various contact t_,pes.,,, Contact Structure Measured Si Dead La),er _}.tm) Boron imp_(p+) 0.46 Gold (1>)') 0.09 Nickel (p+) 0.08 Aluminum (n+) < 0 02 V ACKNOWLEDGMENTS We would like to acknowledge the many contributions mad_ by the other members of the Semiconductor Spec- trometer Team at Lawrence Berkeley Laboratory including R.H. Pehl, C.P. Cork, D.F. Malone, Mj. Pollard, A. Jue, 5 </X-RRY > i._BiiqS N-TYPE I SURFSCE _p N+ / ITO FET GSTE Fig. 7: Field pattern of conventional detector. </X-RRY > i._BiiqS N-TYPE I SURFSCE _p N+ / ITO FET GSTE Fig. 7: Field pattern of conventional detector. e"X-RRY l+BIFqS SLK_FIcENTYPE__s+ _p p+ 1 tTO FET GFITE Fig. 8: Field patmm of reconfigured electrode detector. E. Converse, Y. Wong, H. Sommer and J.T. Walton, in .addition to the assistance of R.M. Ferment in the </X-RRY > i._BiiqS N-TYPE I SURFSCE _p N+ / ITO FET GSTE The authors wish to thank the "BBXRT" team at God- dard Space Flight Center for bringing this challenging proj- ect to our attention and for providing partial funding, This work was supported by the Director's Office of En- ergy Research, Office of Health and Environmental Re- search, U.S. Department of Energy under Contract No. DE- AC03-76SF00098. Reference to a company or product name does not imply approval or recommendation of the product by the University of California or the U.S, Depart- ment of Energy to the exclusion of others that may be suitable. VI. REFERENCES VI. REFERENCES [1] V. Rade,ka, State of the Art of Low Noise Amplifiers for Seraiconductor Radiation Detectors, Intl. Symp. on Nv,clear Electronics, Versailles, France 46:!-28 (1968) [2] D. McCammon, private communication (1989). [2] D. McCammon, private communication (1989). Fig. 7: Field pattern of conventional detector. Fig. 7: Field pattern of conventional detector. [3] J. Howes, private communication (1983), [3] J. Howes, private communication (1983), [3] J. Howes, private communication (1983), [4] D.A. Landis, F.S. Goulding, R.H. Pehl and J.T. Walton, Pulsed Feedback Techniques for Semicon- ductor Detector Radiation SpeCtrometers, IEEE Trans, Nucl Sci., NS-18. No.l, 115-124 (1971). e"X-RRY l+BIFqS SLK_FIcENTYPE__s+ _p p+ 1 tTO FET GFITE Fig. 8: Field patmm of reconfigured electrode detector. E. Converse, Y. Wong, H. Sommer and J.T. Walton, in .addition to the assistance of R.M. Ferment in the manuscript preparation and E. Polonsky for his technical illustration. A special note of thanks is extended to R.H. Pehl who has asked many insightful questions. e"X-RRY l+BIFqS SLK_FIcENTYPE__s+ _p p+ 1 tTO FET GFITE [5] D.A. Landis, C.P. Cork, N.W. Madden and F.S. Goulding, Transistor Reset Preamplifier for High Rate, High Resolution Spectroscopy, IEEE Trans. Nucl, Sci., N_._S-29,No.I, 619-624 (1982). [6] F.S. Goulding, D.A. Landis and N.W. Madden, Design Philosophy for High Resolution Rate and No.l, 301-310 (1983), Throughput Systems, [EEE Trans. Nucl, Sci., [7] V. Radeka, Charge Amplification Without Charge Leak Resistor, IEEE Trans. Nucl. Sci., ._ No.3, 433-439 (1970). [8] J, Llacer, E.E. Hailer and R,C. Cordi, Entrance Win- dows in Germanium Low-Energy X-Ray Detectors, IEEE Trans. Nucl Sci., NS-24. No.l, 53-60 (1977). [9] F.S. Goulding, Some Aspects of Detectors and Elec- tronics for X-Ray Fluorescence Analysis, Nucl. Instr. Meth., 142, 213-223 (1977). Fig. 8: Field patmm of reconfigured electrode detector. [10] J.M. Jaklevic, J.T. Walton, R.E. McMurmy, Jr., N.W. Madden and F.S. Goulding, Semiconductor Detecter Performance for Low-Energy X-Rays, Nucl. Instr. Meth., _ 598..601 (1988). E. Converse, Y. Wong, H. Sommer and J.T. Walton, in .addition to the assistance of R.M. Ferment in the manuscript preparation and E. Polonsky for his technical illustration. A special note of thanks is extended to R.H. Pehl who has asked many insightful questions. i	30,550
https://openalex.org/W2158850967	OpenAlex	Open Science	CC-By	2,014	Serum Samples That Have Been Stored Long-Term (>10 Years) Can Be Used as a Suitable Data Source for Developing Cardiovascular Risk Prediction Models in Large Observational Rheumatoid Arthritis Cohorts	Elke Arts	English	Spoken	6,817	12,663	Version of the following full text: Publisher’s version Downloaded from: http://hdl.handle.net/2066/137571 Download date: 2024-10-24 Serum samples that have been stored long-term (>10 years) can be used as a suitable data source for developing cardiovascular risk prediction models in large observational rheumatoid arthritis cohorts Arts, E.E.A.; Popa, C.D.; Smith, J.P.; Arntz, O.J.; Loo, F.A.J. van de; Donders, A.R.T.; Semb, A.G.; Kitas, G.D.; Riel, P.L.C.M. van; Fransen, J. 2014, Article / Letter to editor (Biomed Research International, 2014, (2014), pp. 930925) Doi link to publisher: https://doi.org/10.1155/2014/930925 Note: To cite this publication please use the final published version (if applicable). Hindawi Publishing Corporation BioMed Research International Volume 2014, Article ID 930925, 8 pages http://dx.doi.org/10.1155/2014/930925 Hindawi Publishing Corporation BioMed Research International Volume 2014, Article ID 930925, 8 pages http://dx.doi.org/10.1155/2014/930925 Elke E. A. Arts,1 Calin D. Popa,1 Jacqueline P. Smith,2 Onno J. Arntz,1 Fons A. van de Loo,1 Rogier Donders,3 Anne Grete P. Semb,4 George D. Kitas,2 Piet L. C. M. van Riel,1 and Jaap Fransen1 Department of Rheumatology, Radboud University Medical Centre, Nijmegen, The Netherlands Department of Rheumatology, Dudley Group NHS Foundation Trust, Dudley, UK 3 Department of Epidemiology, Biostatistics and Health Technology Assessment, Radboud University Medical Centre, Nijmegen, The Netherlands h 4Department of Rheumatology, Diakonhjemmet Hospital, Oslo, Norway Correspondence should be addressed to Elke E. A. Arts; elke.arts@radboudumc.nl Correspondence should be addressed to Elke E. A. Arts; elke.arts@radboudumc.nl Received 31 July 2014; Accepted 22 August 2014; Published 11 September 2014 Received 31 July 2014; Accepted 22 August 2014; Published 11 September 2014 Academic Editor: Miguel Angel Gonzalez-Gay Academic Editor: Miguel Angel Gonzalez-Gay Copyright © 2014 Elke E. A. Arts 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. Objective. There is an unmet need for a specific cardiovascular risk (CV) algorithm for rheumatoid arthritis (RA) patients. Lipoprotein data are often not available in RA cohorts but could be obtained from frozen blood samples. The objective of this study was to estimate the storage effect on lipoproteins in long-term (>10 years) frozen serum samples. Methods. Data were used from an inception RA cohort. Multiple serum samples from 152 patients were analyzed for lipoproteins, being frozen for 1–26 years at −20∘C. Storage effect on lipoproteins was estimated using longitudinal regression analyses and a lipid decay correction factor was developed. Clinical impact of the storage effect on lipoproteins was assessed by calculating the number of patients reclassified to another CV risk group according to the SCORE risk calculator after applying the decay correction factor. Results. There was a significant effect of storage time on total cholesterol (TC) (P < 0.001) and high density lipoprotein cholesterol (HDL-c) levels (P < 0.001), not LDL-c (P = 0.83). The lipid decay correction factor was 0.03 mmol/L and 0.024 mmol/L per additional year of storage for TC and HDL-c, respectively. The TC : HDL ratio decreased after correction for storage effect. After correction, only 5% of patients were reclassified to another CV risk group. Conclusion. A modest storage decay effect on lipoproteins was found that is unlikely to significantly affect CV risk stratification. Serum samples that have been stored long-term (>10 years) can be used to obtain valid lipid levels for developing CV risk prediction models in RA cohorts, even without applying a decay correction factor. 1. Introduction prevalent in RA patients compared to the general population [4–7]. In comparison to healthy controls and RA patients in remission, RA patients with active disease seem to have less stable plaques that are more vulnerable to rupture, which increases the probability of a CV event. Considering the increased risk of CV disease, prevention of CV disease is important. According to international guidelines this includes adequate risk assessment using a CV risk algorithm. In the general population, several algorithms for the Cardiovascular (CV) risk is increased in patients with rheumatoid arthritis (RA) [1]. Cardiovascular diseases account for 50% of all excess mortality in RA patients [1]. RA itself as a chronic inflammatory condition may increase CV risk. Also, studies have shown that inflammation may modulate traditional CV risk factors [2, 3]. Atherosclerotic plaques in the carotid artery appear more severe and BioMed Research International 2 1 2 3 4 5 Storage time 1985 1990 1995 2000 2005 2010 Subcohort 1 inclusion: 1985–1989 Subcohort 2 inclusion: 1990–1994 Subcohort 3 inclusion: 1995–1999 Subcohort 4 inclusion: 2000–2004 Subcohort 5 inclusion: 2005–2009 Figure 1: Storage and blood sampling times for the 5 subcohorts. Symbols represent the time points of the included selection of stored serum, during follow-up (baseline and year 1, 2, 3, 5, 7 and 10). 1 2 3 4 5 Storage time 1985 1990 1995 2000 2005 2010 prediction of CV risk are available, such as the systematic coronary risk evaluation (SCORE) and the Framingham risk score [9, 10]. It has recently been reported that both the SCORE and Framingham risk score algorithm provide suboptimal CV risk estimates in patients with RA [11, 12]. To improve CV risk assessment in RA patients, disease specific risk factors may be required such as RA disease activity. Also, other cardiovascular-related parameters not incorporated in the present algorithms, such as carotid artery intima-media index (cIMT), the presence of plaques in these patients, or certain genetic markers associated with CV risk factors, might also be taken into consideration [13–17]. There is an unmet need for development of a RA specific CV risk calculator. In order to develop and validate appropriate CV risk models for patients with RA, it would be advantageous to use data from existing cohorts with long follow-up. In existing cohorts of RA patients, lipid levels usually are not determined at baseline. Long-term storage may lead to degradation of cholesterol, that is, a lipid decay. Arnhem Nijmegen and informed consent was acquired from all participants. 2.2. Patients. Inclusion criteria for the early RA cohort were fulfillment of the 1987 ACR classification criteria for RA, dis- ease duration less than one year, and being DMARD (disease- modifying antirheumatic drug) na¨ıve. From this cohort, we selected at random 150 RA patients from the inception cohort using computer generated random numbers, to obtain 30 samples per subcohort. Within a timeframe of 1-2 years of storage, no change to moderate decreases in lipid levels have been reported [18– 20], as well an increase of high density lipoprotein cholesterol (HDL-c) levels [21, 22]. Lipid decay seems to be smaller at lower temperatures [18–20]. This has led to the hypothesis that the HDL concentration influences the effect of storage on lipoproteins [23]. Overall, total cholesterol (TC) and triglyceride (TG) levels seems to decay less when stored than HDL-c levels [24, 25]. One study has investigated the effect of long-term storage on cholesterol levels for up to seven years of storage. A significant mean decrease of 2.0% per year storage in TC levels and a nonsignificant average 1.3% decrease per year storage in HDL-c levels were reported [26]. To our knowledge, the effect of longer storage (10 years) on serum cholesterol levels has not been investigated. Although deterioration of cholesterol content in stored serum samples can be expected, the magnitude of this effect after long periods of time is unknown. The objective of this study is to estimate the long time storage decay effect on TC and HDL-c levels in frozen serum samples of RA patients and to evaluate the clinical effect of the decay in CV risk models. 2.3. Serum Samples. During follow-up, nonfasting blood samples were drawn annually by a trained nurse. Approxi- mately 400 mL of serum was stored from each sample and divided into four separate vials. The samples were initially stored at −20∘C. In 2008, all samples were transferred to storage facilities at −80∘C. Blood samples collected from 2008 and thereafter were stored directly at −80∘C. Blood samples obtained before 2007 were stored in 1.5 mL Eppendorf vials and samples obtained after 2007 were stored using Greiner “pp Cryovials.” Serum samples that were taken at baseline and during follow-up (at 1, 2, 3, 5, 7, and 10 years) were extracted from storage in January 2012. 1. Introduction Consequently, if lipids were to be measured in samples that have been stored for longer periods of time, cholesterol levels could be underestimated. Furthermore, if these lipid levels were used in CV risk algorithms, the result may be an underestimation of CV risk. Figure 1: Storage and blood sampling times for the 5 subcohorts. Symbols represent the time points of the included selection of stored serum, during follow-up (baseline and year 1, 2, 3, 5, 7 and 10). Arnhem Nijmegen and informed consent was acquired from all participants. Immediately following this procedure, samples were prepared for cholesterol measure- ments and transported on dry ice to the laboratory facilities of Russells Hall Hospital, Dudley, UK. 2. Methods 2.4. Lipid Measurements. TC concentrations were measured enzymatically by means of the VITROS CHOL slide tech- nique using the Triton X-100 surfactant, which is based on methods described previously [27]. HDL-c was measured using immunoturbidimetry. Low-density-lipoprotein choles- terol (LDL-c) was calculated using Friedewald’s formula [28]. 2.1. Study Design. Serum samples at baseline and at 1, 2, 3, 5, 7, and 10 years of follow-up from patients included in the RA inception cohort of the Radboud University Nijmegen Medical Centre, from 1985 up to 2009 (𝑛= 640), were used for measurements of lipoproteins. To test for a period effect, patients were stratified in five subcohorts according to year of inclusion in the cohort during 1985–1989, 1990–1994, 1995–1999, 2000–2004, and 2005–2009 (Figure 1). The study was approved by the Medical Ethical Committee and CMO 2.5. Statistical Analysis. The primary outcomes were the change in TC and HDL-c levels from baseline to after storage and the secondary outcome was the change in LDL-c level BioMed Research International 3 Table 1: Patient characteristics. 𝑛= 152 Subcohorts 1 (𝑛= 30) 1985–1989 2 (𝑛= 31) 1990–1994 3 (𝑛= 30) 1995–1999 4 (𝑛= 30) 2000–2004 5 (𝑛= 31) 2005–2009 Agea 51 ± 14.4 50 ± 13.5 52 ± 14.5 58 ± 12.1 59 ± 12.6 Femaleb 16 (53.3) 19 (61.3) 17 (56.7) 25 (83.3) 21 (67.7) RF positiveb 26 (86.7) 23 (74.2) 25 (83.3) 23 (76.7) 25 (80.6) DAS28a 5.6 ± 1.2 5.3 ± 1.4 4.8 ± 1.6 4.9 ± 1.0 5.0 ± 1.2 BMIa 26 ± 3.8 27.1 ± 4.6 25.6 ± 3.3 26.2 ± 3.0 26.6 ± 6.8 Smokersb 14 (46.7) 6 (19.4) 10 (33.3) 9 (30.0) 9 (29.0) Statin useb 0 (0.0) 0 (0.0) 2 (6.7) 2 (6.7) 9 (29.0) Glucocorticosteroid useb 3 (10.0) 3 (9.7) 5 (16.7) 17 (56.7) 17 (54.8) Baseline lipoprotein levels TC (mmol/L)a 4.2 ± 1.2 4.5 ± 1.5 5.4 ± 1.0 5.3 ± 1.2 5.2 ± 1.2 HDL-c (mmol/L)a 0.6 ± 0.2 0.9 ± 0.3 1.0 ± 0.3 1.1 ± 0.2 1.2 ± 0.4 TC : HDL-ca 7.2 ± 1.9 5.5 ± 1.6 5.9 ± 1.4 5.1 ± 1.2 4.5 ± 1.2 Treatment during follow-up B-DMARDSb 9 (30.0) 13 (41.9) 10 (33.3) 17 (56.7) 3 (9.7) (i) Abbreviations: RF, rheumatoid factor; DAS28; disease activity index 28 joints, BMI: body mass index, TC, total cholesterol; HDL-c, high-density-lipoprotein cholesterol, B-DMARDS; biological disease modifying anti-rheumatic drugs. 2. Methods (ii) aData presented as mean ± SD, bData presented as 𝑛(%). In order to evaluate the clinical effect of the decay in CV risk models, reclassification across CV risk groups before and after correction was calculated. The SCORE risk algorithm was used to quantify the 10-year risk of CVD with and without correction for the storage decay effect. The CV risk was calculated with and without correction for the storage decay effect on lipids for all 1050 patients from the RA inception cohort. Reclassification of patients across CV risk groups (low <10%, intermediate 10–20%, and high <20%) was calculated. If CV risk of a patient exceeds 10%, primary prevention in the form of lifestyle changes or medical treatment is indicated according to European guidelines for CV prevention [29]. from baseline to after storage. To test for a period effect, a longitudinal regression analysis was used that corrects for repeated measurements within patients. Lipid level (TC and HDL-c) was the dependent variable and follow-up time; subcohort (1985–1989, 1990–1994, etc.) and an interaction term between follow-up time and subcohort were the main independent variables. As the course of cholesterol levels over follow-up time was nonlinear, a quadratic time term (time2) was included. To test for a period effect in the course of cholesterol levels, the interaction between subcohort and follow-up time was evaluated. Several variables were considered as potential confounders: age, gender, statin use at baseline, BMI, smoking, blood pressure, DAS28 score, rheumatoid factor positivity, and glucocorticosteroid use. Variables were considered confounders if their addition to the model led to a >10% change in one of the subcohort follow-up time effects. 3. Results Subcohort 1.50 1.00 0.50 0.00 1 (1985–1989) 2 (1990–1994) 4 (2000–2004) 5 (2005–2009) Follow-up (years) Mean HDL-cholesterol (mmoI/L) 0 1 2 3 5 7 10 6.50 6.00 5.50 5.00 4.50 4.00 3.50 3.00 0 1 2 3 5 7 10 Time points during follow-up (years) Mean total cholesterol (mmol/L) (a) (b) Figure 2: Lipoprotein levels measured in stored serum samples in the various subcohorts. Total cholesterol (a) and HDL-c (b) are depicted on the 𝑦-axis. Samples taken at the most recent follow-up moment in time, time point “10” on the 𝑥-axis, have the shortest follow-up time and samples taken at baseline (time point “0”) have been stored the longest. disproportionate decay effect on TC and HDL-c. This lipid decay was estimated to be linear. A lipid decay correction factor was calculated to be [𝑦= 𝛽observed + (𝛽Chol ∗𝑡)] 0.03 mmol/L for TC and 0.024 mmol/L for HDL-c. Figure 3 illustrates the differences in the course of unadjusted and adjusted cholesterol levels. (not shown) revealed a significant interaction effect between subcohort and follow-up time for TC and LDL-c (𝑃= 0.02 and 𝑃= 0.01, resp.). Overall, the course of the various lipoprotein levels over time was not significantly different between subcohorts after adjustment for confounders (age, gender, and BMI) with 𝑃= 0.09, 𝑃= 0.05, and 𝑃= 0.18 for TC, LDL-c, and HDL-c, respectively. Rheumatoid factor and DAS28 were not confounders after these adjustments.i 3.4. Clinical Impact of the Storage Decay Effect on Lipids. The storage decay effect of lipids during storage affects the TC : HDL-c ratio. This ratio is used when calculating the 10-year CV risk of individual patients in a clinical setting. Patients are then categorized as either “low” risk (<10% 10- year risk of a CV event), “intermediate” risk (10–20% 10-year risk of a CV event), or “high” risk (>20% 10-year risk of a CV event). As a result of the storage decay effect, the TC : HDL- c ratio calculated using the measured lipoprotein levels decreases as storage time increases. To better approximate the lipoprotein levels at the time the serum sample was taken, and the lipid decay correction factor was estimated. Correction for the storage decay effect will yield a higher (improved) TC : HDL ratio which reduces the calculated CV risk. After applying the storage correction factor, the TC : HDL-c ratio decreased. 3. Results 3.1. Patients. One hundred and fifty two patients were included, evenly distributed across the 5 subcohorts (Table 1), with storage times ranging from 1 to 26 years. Samples from the oldest subcohorts comprised the longest storage times. Serum samples from seven time points (0, 1, 2, 3, 5, 7, and 10 years) were analyzed if available, yielding a total of 971 samples. Age, gender, rheumatoid factor (RF) positivity, disease activity index 28 joints (DAS28), use of statins, and glucocorticosteroids appeared to show trends over time (Table 1). f For the development of a correction factor for the storage decay effect, linear mixed models were used, with cholesterol level as the dependent variable, storage time as primary independent variable, and the same confounders as in the analyses described previously. Storage time was calculated by subtracting the baseline date (date the blood samples were first frozen) from the date of serum analysis. The storage decay correction factor developed to adjust the TC, HDL- c, and LDL-c levels was defined as the estimated change in mmol/L cholesterol (𝛽Chol) per additional unit of storage time (years) multiplied by the number of storage years (𝑡) of a particular sample. When added to the measured cholesterol level (𝛽observed), it gives an estimate of the “original” cholesterol value (𝑦). The lipid storage decay factor is therefore 𝑦= 𝛽observed + (𝛽Chol ∗𝑡). 3.2. Differences between Subcohorts. Lipid levels measured at baseline are presented in Table 1. Lipid levels tended to be lowest in the subcohorts (Figure 2) that had the longest storage time and there appeared to be a nonlinear course of lipid levels during storage time. The unadjusted results BioMed Research International 4 Subcohort 1 (1985–1989) 2 (1990–1994) 3 (1995–1999) 4 (2000–2004) 5 (2005–2009) 6.50 6.00 5.50 5.00 4.50 4.00 3.50 3.00 0 1 2 3 5 7 10 Time points during follow-up (years) Mean total cholesterol (mmol/L) (a) Subcohort 1.50 1.00 0.50 0.00 1 (1985–1989) 2 (1990–1994) 3 (1995–1999) 4 (2000–2004) 5 (2005–2009) Follow-up (years) Mean HDL-cholesterol (mmoI/L) 0 1 2 3 5 7 10 (b) Figure 2: Lipoprotein levels measured in stored serum samples in the various subcohorts. Total cholesterol (a) and HDL-c (b) are depicted on the 𝑦-axis. Samples taken at the most recent follow-up moment in time, time point “10” on the 𝑥-axis, have the shortest follow-up time and samples taken at baseline (time point “0”) have been stored the longest. 3. Results Estimate SE 𝑃-value 95% CI Lower Upper Constant 3.408 0.389 <0.0001 2.65 4.17 Storage time −0.030 0.008 <0.0001 −0.045 −0.015 Time within patients (follow-up) 0.012 0.003 <0.0001 0.001 0.018 Time2 (follow-up) −0.0001 0.00003 <0.0001 −0.0002 −0.0001 Gender 0.023 0.097 0.810 −0.167 −0.213 Age 0.016 0.004 <0.0001 0.087 0.023 Statin use at baseline 0.888 0.188 <0.0001 0.519 1.256 BMI 0.011 0.011 0.343 −0.011 0.033 Glucocorticosteroid use at baseline 0.239 0.115 0.038 0.013 0.465 (i) SE, standard error; BMI, body mass index; and 95% CI: 95% confidence interval. (ii) Data are adjusted for age, gender, statin use at baseline, BMI, and glucocorticosteroid use at baseline. (iii) Time2 is a quadratic term that was included due to the nonlinear course of cholesterol levels over follow-up time, and this variable also represents time within patients (follow-up time). i (ii) Data are adjusted for age, gender, statin use at baseline, BMI, and glucocorticosteroid use at baseline. (iii) Ti 2 i d i h i l d d d h li f h l l l l f ll (iii) Time2 is a quadratic term that was included due to the nonlinear course of cholesterol levels over follow-up time, and this variable also represents time within patients (follow-up time). Table 3: Effect of storage time (years) on HDL-c levels, adjusted results. Estimate SE 𝑃value 95% CI Lower Upper Constant 1.247 0.056 <0.0001 1.136 1.358 Storage time −0.024 0.002 <0.0001 −0.027 −0.021 Time within patients (follow-up) 0.003 0.001 <0.0001 0.001 0.004 Time within patients2 (follow-up) −0.00003 0.00001 <0.0001 −0.00004 −0.00002 Age 0.003 0.001 0.001 0.001 0.004 Gender −0.074 0.021 0.001 −0.117 −0.031 (i) SE, standard error; 95% CI, 95% confidence interval. (ii) Data are adjusted for age, gender, and time within patients2. Table 3: Effect of storage time (years) on HDL-c levels, adjusted results. to distorted risk predictions if these values were used in a CV risk model. However, this does not seem to be the case. Clinical impact of the storage decay effect appears to be minimal, with only a small number of patients (5%) moving from groups indicated for primary CV prevention (either the intermediate or high risk) to the low risk group after application of a correction factor. Although, on an individual level, changes in absolute lipid levels may be relevant, these changes do not appear to significantly affect cardiovascular risk predictions. 3. Results Before correction, TC : HDL ratio was classified as high in 75% of patients, compared to 54% after correction (not shown). After correction, patients moved to lower CV risk groups, as the TC : HDL-c ratio decreases after correction for the storage decay effect (Table 4). Before correction, most patients were categorized in the “low” CV risk group (<10%) and 53 patients were reclassified from the intermediate and high risk groups to this group, totaling 552 patients (an 11% increase) after correction. The intermediate (10–20%) and high (>20%) CV risk groups decreased in size by 8% and 11%, t When looking specifically at the oldest and most recent cohort (the two extremes in terms of storage time), lipid levels in the oldest cohort were systematically lower than lipid levers in the most recent subcohort; a statistically significant difference for TC and LDL-c (𝑃= 0.04 and 𝑃= 0.03) and a nonsignificant difference for HDL-c (𝑃= 0.25) were found after correction for confounders. 3.3. Storage Time. There was a significant decay effect of storage time on TC and HDL-c levels (𝑃< 0.001) (Tables 2 and 3). No effect of storage time was found for LDL-c levels (𝑃= 0.83, data not shown). For the analysis for TC and LDL- c, age, gender, BMI, statin use, and glucocorticosteroid use at baseline were confounders and adjusted for. For HDL-c, a model with adjustment for age and gender sufficed.if fi As a significant decay effect was found for TC and HDL-c, a correction factor was estimated. As storage time increased, a decrease (95% CI) was observed, −0.03 mmol/L (−0.045– −0.015) for TC and −0.024 mmol/L (−0.027–−0.021) for HDL- c per year of increasing storage time. The decrease in HDL-c levels was relatively larger (considering the range) than the decrease in TC levels at same length of storage. The TC/HDL ratio calculated for the same sample will therefore become higher as a direct result of increasing storage time and the BioMed Research International 5 Table 2: Effect of storage time (years) on TC levels. 3. Results Therefore, it may not be necessary to use a correction factor when samples have been stored under stable conditions. respectively. Overall, in this cohort of 1050 patients, 53 (5%) patients changed CV risk groups according to SCORE 10-year risk predictions for CVD. 4. Discussion Hence a different approach BioMed Research International 6 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 1 0 1 2 3 5 7 10 (a) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 2 0 1 2 3 5 7 10 (b) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 3 0 1 2 3 5 7 10 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 4 0 1 2 3 5 7 10 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 1 0 1 2 3 5 7 10 (a) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 2 0 1 2 3 5 7 10 (b) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 3 0 1 2 3 5 7 10 (c) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 4 0 1 2 3 5 7 10 (d) 0 1 2 3 5 7 10 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 CholmmolL.1: chol (mmol/L) HDLmmolL.1: HDL (mmoI/L) LDLmmolL.1 TC adjusted HDL adjusted LDL adjusted Error bars: 95% CI Subcohort 5 (e) Figure 3: Lipoprotein levels measured over time in the various subcohorts before and after correction with the lipid decay factor. Mean TC and HLD-c levels are depicted on the 𝑦-axis and follow-up time on the 𝑥-axis. Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 2 0 1 2 3 5 7 10 (b) Follow-up Mean cholesterol (mmol/L) 6 4 2 0 1 0 1 2 3 5 7 10 Subcohort Follow-up (c) (d) 0 1 2 3 5 7 10 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 CholmmolL.1: chol (mmol/L) HDLmmolL.1: HDL (mmoI/L) LDLmmolL.1 TC adjusted HDL adjusted LDL adjusted Error bars: 95% CI Subcohort 5 (e) gure 3: Lipoprotein levels measured over time in the various subcohorts before and after correction with the lipid decay factor. Mean TC d HLD-c levels are depicted on the 𝑦-axis and follow-up time on the 𝑥-axis. (c) (d) 0 1 2 3 5 7 10 Follow-up Mean cholesterol (mmol/L) 6 4 2 0 Subcohort 5 (c) (d) (e) Figure 3: Lipoprotein levels measured over time in the various subcohorts before and after correction with the lipid decay factor. Mean TC and HLD-c levels are depicted on the 𝑦-axis and follow-up time on the 𝑥-axis. Table 4: CV risk reclassification. 4. Discussion Our study is the first to investigate the suitability of choles- terol levels obtained in serum samples that have been stored for more than 10 years and to evaluate if this storage decay effect is clinically important for CV risk evaluation in RA patients using available CV risk calculators. We show that a significant but modest storage decay effect on cholesterol levels occurs over time. The magnitude of this effect per addi- tional year of storage is relatively small. Correcting factors to adjust for this effect were calculated, 0.03 mmol/L for TC and 0.024 mmol/L for HDL-c, per additional year of storage. Interestingly, the change in TC and HDL-c per additional year of storage in this cohort is similar, which means that the impact of this storage decay effect will be greater for HDL- c, as the range of HDL-c levels is much smaller compared to TC levels. The decay effect of TC levels reported in this study is less steep than the decrease in HDL-c. Therefore, the observed TC : HDL ratio may become increased and lead to false-higher CV risk predictions. In particular, lipid measurements in samples that have undergone long-term storage could be inaccurate, which would eventually lead Determining whether long-term storage affects the integrity of lipid samples requires analysis of long-term data. Ideally one would compare values determined in the past, directly after acquiring the blood sample (“original” values), with values determined from stored samples (“observed” val- ues) within patients. The differences between “original” val- ues and “observed” values provide an immediate indication of the storage effect. Several studies have used this approach but in serum samples that were stored for relatively short periods of 1-2 years [19, 20, 24, 25]. However, measurements of lipids directly after blood sampling are not always available in RA cohorts. This may be due to the fact that the awareness of the increased risk of CVD gained particular interest long after the start of RA cohorts. 4. Discussion CV risk Before correction for storage decay 𝑛(%) After correction for storage decay 𝑛(%) Reclassified 𝑁= 53 Low (<10%) 499 (48%) 552 (53%) +53 (11%) Intermediate (10–20%) 178 (17%) 164 (16%) −14 (8%) High (>20%) 373 (35%) 334 (31%) −39 (11%) Total 𝑛(%) 1050 (100%) 1050 (100%) +53 (5%) (i) CV risk, cardiovascular risk. (ii) Number of patients reclassified according to the SCORE CV risk categories for the 10-year risk of a CV event; (low; <10%, intermediate; 10–20%, and high; >20%) before and after correction for storage decay effect on TC and HDL-c. Table 4: CV risk reclassification. (i) CV risk, cardiovascular risk. (ii) Number of patients reclassified according to the SCORE CV risk categories for the 10-year risk of a CV event; (low; <10%, intermediate; 10–20%, and high; >20%) before and after correction for storage decay effect on TC and HDL-c. according to the SCORE CV risk categories for the 10-year risk of a CV event; (low; <10%, intermediate; 10–20%, and high; for storage decay effect on TC and HDL-c. would reflect the storage effect [26]. However, in a long- running cohort that includes a lengthy follow-up of more than 10 years, patients could also be systematically different between different time-periods. For example, patients that were included more recently are probably more likely to use statins at the time the serum samples were taken than patients that were included 25 years ago. In this cohort, cholesterol levels from samples that were stored the shortest could therefore be systematically lower than samples that were is required to assess the storage decay effect on lipid levels in these cohorts. In a study that investigated the effect of long-term storage (7 years) [26], the group means of lipids for pairs of serial specimens that were taken at 6- and 12- month intervals were compared. It was assumed that in the absence of a storage effect the variation in group means would reflect only normal biological variation, which would not lead to a systematic downward or upward effect in the group mean cholesterol levels. Subsequently, any observed changes 7 BioMed Research International recommended to employ the method presented in this study in other long-term cohorts. stored the longest, without involvement of a storage effect. Such a period effect could lead to biased results. References [1] J. A. Avina-Zubieta, J. Thomas, M. Sadatsafavi, A. J. Lehman, and D. Lacaille, “Risk of incident cardiovascular events in patients with rheumatoid arthritis: a meta-analysis of observa- tional studies,” Annals of the Rheumatic Diseases, vol. 71, no. 9, pp. 1690–1697, 2012. [2] P. H. Dessein, B. I. Joffe, and S. Singh, “Biomarkers of endothe- lial dysfunction, cardiovascular risk factors and atherosclerosis in rheumatoid arthritis,” Arthritis Research & Therapy, vol. 7, no. 3, p. R634, 2005. [3] P. H. Dessein, A. E. Stanwix, and B. I. Joffe, “Cardiovascular risk in rheumatoid arthritis versus osteoarthritis: acute phase response related decreased insulin sensitivity and high-density lipoprotein cholesterol as well as clustering of metabolic syn- drome features in rheumatoid arthritis,” Arthritis Research, vol. 4, no. 5, p. R5, 2002. [4] H. Kobayashi, J. T. Giles, J. F. Polak et al., “Increased prevalence of carotid artery atherosclerosis in rheumatoid arthritis is artery-specific,” The Journal of Rheumatology, vol. 37, no. 4, pp. 730–739, 2010. p In conclusion, the results of this study show that modest decay of lipid levels in serum samples should be expected during long-term (>10 years) storage. Using the method proposed here, a correction factor can be formulated to adjust for this storage decay effect. However as the clinical impact on CV predictions appears minimal, extensive adjustment may not be necessary to obtain valid lipid levels. Hence, stored serum samples, even when stored for long periods of time, are a valuable source of data. These data can be of particular importance for studies in RA cohorts involving long-term outcomes such as cardiovascular disease. In addition, it may be financially attractive to minimize the analysis period by performing cholesterol measurements in stored serum samples all at once at the end of a long-term study with multiple samples taken over long periods of time. It is [5] M.-C. Aubry, H. Maradit-Kremers, M. S. Reinalda, C. S. Crowson, W. D. Edwards, and S. E. Gabriel, “Differences in atherosclerotic coronary heart disease between subjects with and without rheumatoid arthritis,” The Journal of Rheumatology, vol. 34, no. 5, pp. 937–942, 2007. [6] C. Gonzalez-Juanatey, J. Llorca, A. Testa, J. Revuelta, C. Garcia-Porrua, and M. A. Gonzalez-Gay, “Increased prevalence of severe subclinical atherosclerotic findings in long-term treated rheumatoid arthritis patients without clinically evident atherosclerotic disease,” Medicine, vol. 82, no. 6, pp. 407–413, 2003. [7] G. A. Karpouzas, J. Malpeso, T. Y. Conflict of Interests The authors declare that there is no conflict of interests regarding the publication of this paper. Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. Abbreviations CV: Cardiovascular SCORE: Systematic coronary risk evaluation TC: Total cholesterol HDL-c: High density lipoprotein cholesterol TG: Triglycerids DMARD: Disease-modifying antirheumatic drug LDL-c: Low-density lipoprotein cholesterol RF: Rheumatoid factor TC : HDL: Total cholesterol : high density lipoprotein cholesterol ratio. y pp This study has limitations. The storage effect was esti- mated without knowing “original” values for comparison. The correction factor was directly derived from the regres- sion coefficient of storage time. It was assumed that after correction for repeated measures and for confounders any observed change is attributable to the storage effect. Biologi- cal variation of lipid levels within a subject could potentially contribute to the found decay effect over time. However, several (𝑛= 7) measurements per patient were included, in the total group of 152 patients. Serum samples had been stored under similar condition, albeit in different storage facilities. It was considered unlikely that any variation, either due to biological variation within patients or due to differ- ing storage circumstances, would contribute to an overall, systemic trend. It has been previously suggested that such variability in measured cholesterol levels is unpredictable and often a wide variation in both directions (±20%) is reported [30, 31]. In the model that was formulated, the most important confounders were dealt with, but the existence of other confounding factors cannot be excluded. Storage conditions, varying temperatures during storage, and number of times the samples were thawed are all factors that could have contributed in affecting serum cholesterol levels. As multiple sets of samples from 152 patients were used, it is considered unlikely that any of these highly variable factors contributed significantly to the systemic decay effect that was found. In addition, modifying the storage temperature from −20∘C to −80∘C for all serum samples in 2008 is a limitation, although the vast majority of the samples (1985–2007) were stored under the same conditions. Storage effects are likely to be even smaller with lower temperatures. 4. Discussion Therefore, we used a method in which the data were grouped according to five different time-periods for analysis, to adjust results for a period effect. Illustratively, statin use at baseline increased from 0% in the first three subcohorts (1985–1999) to 29% in the last cohort (2005–2009). The percentage of smokers decreased from 47% to 29% during that same timeframe. Overall, these subcohorts did not seem to differ significantly, excluding period effect as a confounder. Consequently, a simplified model was developed. The methodology used in this study can also be applied in other cohorts.hhf References Choi et al., “Prevalence, extent and composition of coronary plaque in patients with BioMed Research International 8 [22] J. D. Curb, M. L. Overturf, and R. B. Harrist, “Lability of HDL- cholesterol during serum storage,” Atherosclerosis, vol. 37, no. 4, pp. 641–645, 1980. rheumatoid arthritis without symptoms or prior diagnosis of coronary artery disease,” Annals of the Rheumatic Diseases, 2013. [8] A. G. Semb, S. Rollefstad, S. A. Provan et al., “Carotid plaque characteristics and disease activity in rheumatoid arthritis,” Journal of Rheumatology, vol. 40, no. 4, pp. 359–368, 2013. [23] P. S. Bachorik, R. E. Walker, and P. O. Kwiterovich Jr., “Determi- nation of high density lipoprotein-cholesterol in human plasma stored at -70∘C,” Journal of Lipid Research, vol. 23, no. 8, pp. 1236–1242, 1982. [9] R. M. Conroy, K. Py¨or¨al¨a, A. P. Fitzgerald et al., “Estimation of ten-year risk of fatal cardiovascular disease in Europe: the SCORE project,” European Heart Journal, vol. 24, no. 11, pp. 987– 1003, 2003. [24] Y. M. Stokes, C. E. Salmond, L. M. Carpenter, and T. J. Welby, “Stability of total cholesterol, high-density-lipoprotein cholesterol, and triglycerides in frozen sera,” Clinical Chemistry, vol. 32, no. 6, pp. 995–999, 1986. [10] K. M. Anderson, P. W. F. Wilson, P. M. Odell, and W. B. Kannel, “An updated coronary risk profile. A statement for health professionals,” Circulation, vol. 83, no. 1, pp. 356–362, 1991. [25] H. G. M. Tiedink and M. B. Katan, “Variability in lipoprotein concentrations in serum after prolonged storage at −20∘C,” Clinica Chimica Acta, vol. 180, no. 2, pp. 147–156, 1989. [11] C. S. Crowson, E. L. Matteson, V. L. Roger, T. M. Therneau, and S. E. Gabriel, “Usefulness of risk scores to estimate the risk of cardiovascular disease in patients with rheumatoid arthritis,” The American Journal of Cardiology, vol. 110, no. 3, pp. 420–424, 2012. [26] W. J. Shih, P. S. Bachorik, J. A. Haga, G. L. Myers, and E. A. Stein, “Estimating the long-term effects of storage at -70∘C on cholesterol, triglyceride, and HDL-cholesterol measurements in stored sera,” Clinical Chemistry, vol. 46, no. 3, pp. 351–364, 2000. [27] C. C. Allain, L. S. Poon, C. S. G. Chan, W. Richmond, and P. C. Fu, “Enzymatic determination of total serum cholesterol,” Clinical Chemistry, vol. 20, no. 4, pp. 470–475, 1974. [12] E. E. A. Arts, C. D. Popa, A. A. References den Broeder et al., “Performance of five current risk algorithms in predicting cardiovascular events in patients with early rheumatoid arthritis,” Annals of the Rheumatic Diseases, 2014. [28] W. T. Friedewald, R. I. Levy, and D. S. Fredrickson, “Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge.,” Clinical Chemistry, vol. 18, no. 6, pp. 499–502, 1972. [13] A. Corrales, J. A. Parra, C. Gonz´alez-Juanatey et al., “Car- diovascular risk stratification in rheumatic diseases: carotid ultrasound is more sensitive than Coronary Artery Calcification Score to detect subclinical atherosclerosis in patients with rheumatoid arthritis,” Annals of the Rheumatic Diseases, vol. 72, no. 11, pp. 1764–1770, 2013. [29] J. Perk, G. De Backer, H. Gohlke et al., “European guidelines on cardiovascular disease prevention in clinical practice (version 2012). The fifth joint task force of the European Society of Cardiology and Other Societies on cardiovascular disease prevention in clinical practice (constituted by representatives of nine societies and by invited experts),” European Heart Journal, vol. 33, no. 13, pp. 1635–701, 2012. [14] A. Corrales, C. Gonz´alez-Juanatey, M. E. Peir´o, R. Blanco, J. Llorca, and M. A. Gonz´alez-Gay, “Carotid ultrasound is useful for the cardiovascular risk stratification of patients with rheumatoid arthritis: results of a population-based study,” Annals of the Rheumatic Diseases, vol. 73, no. 4, pp. 722–727, 2014. [30] M. Mogadam, S. W. Ahmed, A. H. Mensch, and I. D. Godwin, “Within-person fluctuations of serum cholesterol and lipopro- teins,” Archives of Internal Medicine, vol. 150, no. 8, pp. 1645– 1648, 1990. [15] L. Rodr´ıguez-Rodr´ıguez, C. Gonz´alez-Juanatey, R. Palomino- Morales et al., “TNFA—308 (rs1800629) polymorphism is asso- ciated with a higher risk of cardiovascular disease in patients with rheumatoid arthritis,” Atherosclerosis, vol. 216, no. 1, pp. 125–130, 2011. [31] S. M. Marcovina, V. P. Gaur, and J. J. Albers, “Biological variability of cholesterol, triglyceride, low- and high- density lipoprotein cholesterol, lipoprotein(a), and apolipoproteins A- I and B,” Clinical Chemistry, vol. 40, no. 4, pp. 574–578, 1994. [16] R. L´opez-Mej´ıas, M. Garc´ıa-Berm´udez, C. Gonz´alez-Juanatey et al., “NFKB1-94ATTG ins/del polymorphism (rs28362491) is associated with cardiovascular disease in patients with rheuma- toid arthritis,” Atherosclerosis, vol. 224, no. 2, pp. 426–429, 2012. [17] R. L´opez-Mej´ıas, C. Gonz´alez-Juanatey, M. Garc´ıa-Berm´udez et al., “The lp13.3 genomic region -rs599839- is associated with endothelial dysfunction in patients with rheumatoid arthritis,” Arthritis Research & Therapy, vol. 14, no. 2, p. R42, 2012. [18] L. L. Bausserman, A. L. References Saritelli, and D. Milosavljevic, “High- density lipoprotein subfractions measured in stored serum,” Clinical Chemistry, vol. 40, no. 9, pp. 1713–1716, 1994. [19] P. N. M. Demacker, A. G. M. Hijmans, D. F. van Sommeren- Zondag, and A. P. Jansen, “Stability of frozen liquid control sera for assay of cholesterol in high-density lipoprotein,” Clinical Chemistry, vol. 28, no. 1, pp. 155–157, 1982. [20] M. N. Nanjee and N. E. Miller, “Evaluation of long-term frozen storage of plasma for measurement of high-density lipoprotein and its subfractions by precipitation,” Clinical Chemistry, vol. 36, no. 5, pp. 783–788, 1990. [21] J. P. Reckless, D. J. Betterridge, P. Wu, B. Payne, and D. J. Galton, “Effect of storage on plasma-lipoproteins,” The Lancet, vol. 2, article 350, 1977. 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https://openalex.org/W1970436382_1	Spanish-Science-Pile	Open Science	Various open science	2,013	Tallado y demarcación: composiciones transpuestas entre el rock de Brasil y de Argentina	None	Spanish	Spoken	7,069	12,459	TALLADO Y DEMARCACIÓN: COMPOSICIONES TRANSPUESTAS ENTRE EL ROCK DE BRASIL Y DE ARGENTINA Adrián Pablo Fanjul Universidade de São Paulo São Paulo, São Paulo, Brasil Resumen: Abordamos, de modo comparativo entre discursividades brasileñas y argentinas, configuraciones para la representación de diversos tipos de entidades personales en la enunciación. Los casos analizados son composiciones poético-musicales, encuadradas en el campo del rock, transpuestas de un país al otro. Trabajamos a partir de un referencial teórico y metodológico ubicado en el análisis discursivo-enunciativo. Observamos las transformaciones que se producen, en el paso de las composiciones de una discursividad y una lengua a la otra, en lo referente al contorno de las figuras y de las voces representadas en la enunciación. Encontramos, en las producciones argentinas, una tendencia al refuerzo de esos contornos y en las producciones brasileñas una mayor permeabilidad con el entorno representado. Palabras-clave: Persona en la enunciación. Discursividad en el rock. Estudios comparados entre Brasil y Argentina. 1 INTRODUCCIÓN Este trabajo se encuadra en un proyecto de investigación más general sobre discursividades en la música urbana de Brasil y de Argentina, específicamente sobre los subcampos que en cada uno de esos dos países han recibido el nombre de “rock nacional”1. La investigación tiene una perspectiva comparativa y se realiza en el marco teórico del análisis  Doutor em Linguística pela UNESP. Email: adrianpf@yahoo.com. El proyecto cuenta con apoyo del Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). En cuanto a la denominación “subcampo”, será desarrollada en la sección siguiente. 1 360 \| Linguagem em (Dis)curso discursivo-enunciativo, articulando conceptos y categorías analíticas que se explicitarán en la segunda sección. Focalizaremos en este artículo la cuestión de las instancias de persona en los diversos niveles de representación que la enunciación y el discurso movilizan, problematizando la delimitación de entidades personales de diferente carácter y alcance involucradas en esos niveles. Lo haremos a partir de una contraposición entre composiciones de rock que fueron adaptadas de un país al otro, transponiendo la letra a la otra lengua. Consideraremos dos temas que sufrieron ese tipo de adaptación, uno de Argentina a Brasil y otro de Brasil a Argentina. “Viernes 3 AM” fue grabada por primera vez en 1979 por el grupo Seru Girán, tercero en la carrera de Charly García, autor del tema. Casi veinte años después la grabó Paralamas do Sucesso en versión de Herbert Vianna para el disco Hey Na Na (1998), manteniendo el título en castellano. La segunda composición es “O tempo não pára”, de Cazuza en su etapa solista, divulgada en el LP en vivo del mismo nombre que apareció en 1988. La entonces naciente banda argentina Bersuit Vergarabat la adaptó y registró en su primer disco, Y punto, de 1991, en versión de Gustavo Cordera y Carlos Martín. Las dos letras y sus versiones están en el anexo ubicado al final del artículo. La sección siguiente presentará nuestro marco teórico y metodológico abordando primero lo relativo a las instancias personales y su relación con la constitución de la subjetividad en el discurso, y posteriormente algunas especificidades sobre la enunciación en el campo estudiado, así como algunas aclaraciones sobre el tipo de hecho de transposición considerado en relación a los estudios sobre traducción. En los ítemes tercero y cuarto realizamos el análisis de los dos casos que nos ocupan, y en el último presentamos conclusiones en relación con las discursividades que han atravesado el campo del rock. 2 PRESUPUESTOS TEÓRICO-METODOLÓGICOS 2.1 SUBJETIVIDAD, SERES DEL ENUNCIADO, PERSONA Para un amplio sector de los estudios discursivos y enunciativos, entre los cuales nos ubicamos, el sujeto del discurso, al menos en uno de Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 361 sus niveles de abordaje, no es una entidad estable a lo largo del enunciado sino un lugar a ser llenado, lugar constituido en el propio proceso discursivo y sin continuidad en el enunciado, ya que este último puede ser atravesado, en diferentes puntos, por diversas posiciones de sujeto. A partir de la postulación que, al terminar la década del 60, Michel Foucault realizara en La arqueología del saber (FOUCAULT, 2007, p. 121-126), se puede ver al sujeto como la posición que se ocupa para formular lo que se formula, una función vacía, determinada por las relaciones del enunciado con otros enunciados en series o dominios delimitables. Desde la corriente francesa del análisis del discurso cuyo principal referente fue Michel Pêcheux, va siendo establecido un diálogo, que podríamos denominar como de adopción polémica, con la teorización de Foucault, explicitado, entre otras obras, en un texto crucial de 1979 (PÊCHEUX, 1997) y, de un modo más convergente, en escritos de los años 80 (PÊCHEUX, 2011)2. En las importantes proyecciones que tuvo la obra de Pêcheux en Brasil, que dieron lugar a una verdadera (y multifacética) escuela de análisis del discurso, la noción de “posición de sujeto”, se halla muy extendida, y con una clara inspiración foucaultiana, como puede verse, por ejemplo, en Orlandi (1988 y 1990) o Guimarães (1989 y 2005). Así, hablar desde una posición de sujeto es hacerlo desde una región del interdiscurso (GUIMARÃES, 2005, p. 30) relacionable con la dimensión ideológica. Otro orden de fenómenos, que entra en relación con el anterior pero es diferente, es el de las voces representadas en el enunciado y por la enunciación, lo que con Guimarães (2005, p. 23) podríamos llamar “agenciamiento enunciativo”. Desde cuando Benveniste (1971, p. 85) situó los “indicios de persona” como centro movilizador de todas las formas de la lengua en el funcionamiento que se pone en marcha por el proceso enunciativo, muchas han sido las propuestas para describir y analizar las diversas figuras personales cuya representación se complejiza por las formas de discurso referido, por la ficcionalización, por los soportes 2 Pêcheux adopta y reformula la noción de “formación discursiva” de Foucault, lo que no quiere decir que comparta integralmente su visión sobre el sujeto. Sobre los modos como la concepción althusseraina de la ideología y del sujeto afectaron la relación de Pêcheux y colaboradores con la teorización de Foucault, recomendamos la lectura de Gregolin (2004) y de la producción última del propio Pêcheux en 1982 (PÊCHEUX, 2002). FANJUL. Tallado y demarcación... 362 \| Linguagem em (Dis)curso mediáticos y por las muchas formas de presencia del discurso “otro” en el enunciado. Destacamos, entre otras, la distinción entre “locutor” y “enunciador” en el abordaje polifónico de la enunciación por Ducrot (2001), así como su recategorización por Guimarães (2005, p. 11-31) en función de “lugares sociales de decir”, relacionados con papeles distinguibles en el funcionamiento social, político y cultural, y “lugares de decir”, en los que se borra la implantación-histórico-social del locutor. Esos lugares se cruzan, de modo a veces poco diferenciado, con las entidades textualmente estabilizadas por la interlocución representada en lo que, con Maingueneau (2001 y 2008) llamaremos “escenografía enunciativa”: locutores, interlocutores representados con diferentes grados de nitidez, personajes. Nuestro trabajo parte de la observación de todas esas figuras representadas en el enunciado y de su modo de articulación y delimitación en torno del decir y sus lugares. Creemos que investigar recurrencias y tendencias para esas configuraciones, en un género o en un campo discursivo en el cual también se puedan reconocer posicionamientos subjetivos recurrentes es revelador, ya que el agenciamiento enunciativo no es una mera forma que vehicule sentidos o ideologías sin afectar su orientación en relación a una memoria del decir. Y si, como en nuestro caso, esa investigación se realiza de modo comparado entre alteridades separadas por la lengua y la nacionalidad, la contraposición de tendencias y predominios para determinadas configuraciones permite alcanzar conclusiones que provean una base sólida, desde la especificidad de las categorías del discurso, para los estudios socioculturales comparados. 2.2 ENUNCIACIÓN EN EL ROCK, CONTORNOS Y NUESTRA INVESTIGACIÓN Como para otros géneros de la cultura de masas o de la llamada “música popular”, creemos que para el rock es adecuada la caracterización que Díaz (2005), basándose en la noción de “campo” en Bourdieu (1989), le atribuye como “subcampo” de la producción y circulación de bienes culturales. Eso significa, entre otras cosas, que también presenta una propiedad fundamental de los campos, como es la disputa de legitimidad Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 363 por el decir. Considerar que haya un decir roquero no debe significar caer en la ilusión monoglósica de pensarlo como el decir de un posicionamiento, de un lugar estable de constitución de sujetos. El subcampo que el rock ha ido conformando está atravesado por las muchas divergencias en su casi medio siglo de existencia, en que varias tradiciones han sido generadas, contestadas, defenestradas y/o recreadas alrededor de los muchos objetos de su discursividad. Una de las recurrencias que encontramos en medio de esa heterogeneidad podemos identificarla con lo que Díaz (2005, p. 154-167) caracteriza como “diferencia disidente”. Desde el punto de vista enunciativo, podemos verla como la escenificación de posicionamientos presentados como opuestos a una racionalidad – que no deja de serle complementaria – expuesta como dominante. La contraposición se realiza de diversas formas: el tópico de llegar a ver lo que otros no ven, la diferenciación respecto de un colectivo o de un contramodelo conforme, la instalación un lugar de decir que puede aparecer como de “saber” o de exaltación de un “no saber”, pero que se proclama como revelador, y varios otros. Las voces de la diferencia disidente están marcadas por la contradicción entre el padecimiento y el goce que tal posicionamiento conlleva. Es muy adecuada al respecto de esa contradicción (aunque no compartamos su generalización a todo el subcampo) esta descripción del crítico literario argentino Monteleone (1992, p. 33) Todo enunciado rocker es reconocido como enunciado de delincuencia. Palabra lacerada de un cuerpo que va en cana, palabra pronunciada en la hora detenida de un toque de queda; pero también palabra gozosa, en la fiesta de la fugaz resistencia y en el ritmo disruptor del cuerpo rocker. Inclusive en composiciones que llevan sus objetos y su mundo representado al plano de lo íntimo, como ocurre por momentos en las que aquí analizaremos, también se manifiestan variantes de ese posicionamiento. En cuanto a la representación de instancias personales en su materialidad verbal, o sea, en su letrística, en el rock ocurren las variadas FANJUL. Tallado y demarcación... 364 \| Linguagem em (Dis)curso posibilidades escenográficas que se observan en otros géneros ligados a la forma canción3: desde soliloquios a grados moderados de ficcionalización o narrativización (pequeñas crónicas), con posibilidad de recibir inflexiones de otros géneros inclusive no artísticos, como proclamas, plegarias y otros. Esas posibilidades dan lugar a la instalación de diversas voces representadas e inclusive, en los casos en que hay un cierto grado de narrativización, de cuasi-personajes. Para todas esas voces, géneros tan performáticos como el rock actualizan notablemente la intercambiabilidad, por incorporación, con el par intérprete/oyente. Aclaramos que consideramos aquí “incorporación” a partir de una de las funciones que atribuye Maingueneau (2001, p. 140) a ese proceso en la relación entre fiador y coenunciador: la constitución de un cuerpo, el de la comunidad imaginaria que participa de la fruición de la obra. Creemos que, por incluir entre sus principales condiciones de circulación el recital y el canto colectivo, el rock es un género en que ese proceso resulta particularmente central. Nos viene interesando, en nuestras investigaciones, la consideración de cómo se representa la relación y la delimitación ser/entorno para las figuras puestas en la escenografía enunciativa y/o para las voces en las que estas toman cuerpo. Creemos que es un factor relacionado con modos de representar la conflictividad, y que por eso es productivo observarlo, en un dominio de géneros y posicionamientos análogos, contraponiendo dos formaciones sociohistóricas diferentes (en nuestro caso, Brasil y Argentina en sus espacios urbanos contemporáneos). Así, en un trabajo anterior (FANJUL, 2005), se indagó, en composiciones del rock de ambos países de las décadas del 80’ y del 90’, casos en que se representa al héroe en relación con un entorno hostil en un conflicto de alcance colectivo, encontrando tendencias diferenciadas para cada país. En las composiciones brasileñas se destacaba una tendencia a representar la inferioridad relativa del héroe y su no control o no percepción del entorno, ambas como refuerzo precisamente de su heroicidad, mientras que en los casos argentinos predominaba una representación del héroe en intento de alcanzar 3 En Tatit (2003, p. 9), la canción es definida como una forma poético-musical caracterizada por una progresión melódica con reiteración. Esa forma puede ser manifestada por géneros muy diversos. Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 365 un dominio perceptivo del entorno y un cierre sobre su propio espacio en un movimiento que llamamos “endocéntrico”. En este trabajo nos ocuparemos de un aspecto de la enunciación relacionado con el ser representado, que analizaremos como contorno tallado por un decir. Usamos la metáfora del tallado precisamente porque de los varios aspectos en que la presencia de un ser puede observarse nos interesa cómo se delimita, en su decir o en el decir que lo construye, de los representados como complementarios de escena, solidarios y/o adversarios y cómo se delimita del entorno en la escenografía. Ese delimitarse tiene que ver con su propia emergencia como figura y su progresiva construcción en el enunciado, ya que como explica Maingueneau (2008, p. 70), la escenografía enunciativa no es un cuadro ya dado e independiente del discurso, sino que la propia enunciación lo va instalando. Por nuestra parte, y análogamente, creemos necesario también considerar que las dimensiones que ese autor propone como cronografía y topografía, así como la corporeidad que los seres adquieren en ellas tienen un carácter representacional, pero de una naturaleza diferente de la representación pictórica o teatral, precisamente porque están hechas de lenguaje verbal. No hay límites previos para su extensión ni planos o líneas previstos para su ocurrencia. La superposición, la indiferenciación, así como la cisión absoluta o la repetición indefinida son posibilidades dadas por el funcionamiento del lenguaje verbal para la representación de todo tiempo, espacio o cuerpo. Por eso lo que denominamos “contorno” no se procesa sólo mediante la descripción o atribución de cualidades y ubicaciones relativas, sino también por las marcas de argumentación y de todo tipo de semantismo. Así, creemos que las muchas formas de determinación, relaciones como adversatividad, causatividad, condición y otras intervienen en el tallado del ser puesto en escenografía cuando lo involucran en diversos roles. En el caso de este trabajo, la contraposición no será entre temas musicales de cada país, sino entre composiciones que fueron adaptados de un país al otro, transponiendo la letra a la otra lengua. Cintrão (2007, p. 131-132) ubica la traducción de canciones en el conjunto de las prácticas que se denominan “traducción subordinada” porque sufren restricciones FANJUL. Tallado y demarcación... 366 \| Linguagem em (Dis)curso como consecuencia de la relación con un lenguaje no verbal, en su caso, el lenguaje musical. A su vez, por la proximidad que esas restricciones conllevan con las de la poesía, evalúa como adecuada la noción de “transposición creativa” propuesta por Jakobson (2001, p. 72) para definir el proceso que de hecho tiene lugar en la versión de la poesía a otra lengua. Creemos que es el tipo de proceso que ocurre en el material que analizamos. A la vez, consideramos que las restricciones que ese proceso conlleva pueden explicar el distanciamiento respecto de la forma presente en la lengua de partida, pero de ninguna manera explican la selección de una forma entre las muchas que cabrían en determinado lugar de un verso cantado en la lengua de llegada. Por lo tanto, creemos que, para cada uno de los lugares de la cadena intradiscursiva en los que vemos un indicio de las delimitaciones de entidades personales que aquí estudiamos, podrían haber ocurrido formas que se integrasen a otro efecto en la construcción de la escena, y no vemos la emergencia de ninguno de los indicadores específicos que observaremos como consecuencia inexorable de las restricciones de la práctica traductora. Intentaremos mostrar en la próxima sección que, en el paso de estas composiciones a su versión otra, los contornos tallados para los seres representados sufren transformaciones. Y que hay cierta regularidad en esas transformaciones, que nos permite percibir la adecuación a lo que aparece como una expectativa en el campo del rock de cada país en relación con los seres y sus entornos. 3 EL TALLADO Y SUS LASCAS “Viernes 3 AM” apareció en el segundo LP de Seru Girán, La grasa de las capitales (1979). En varios aspectos ese disco puede verse como un punto de referencia histórico. En su tema inicial y homónimo introduce, por momentos de modo paródico, ritmos que invitan al movimiento corporal, cosa extremamente infrecuente en el rock argentino de su época, y que prefigura lo que será una fuerte tendencia en los años posteriores. La obra inicia también un viraje fuertemente crítico en relación con el “estado de cosas” en el país, en un momento en que sectores importantes de las clases medias comienzan a manifestar una difusa disconformidad. No alude Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 367 directamente a la dictadura, pero sí al clima de mediocridad reinante en el campo cultural y en los medios. Sin embargo, todavía incluye expresiones de la etapa más lóbrega en la poética de Charly García. Persiste la familiaridad con diferentes formas de la muerte, el tono y la atmósfera melancólica de los mundos creados que una generación naturalizó ya antes de la dictadura, a través de temas como “Rasguña las piedras” (1973), casi todos los del LP Instituciones (1974) o “Nena” (1975, grabado después como “Eiti Leda” en el primer LP de Seru, en 1978). “Viernes 3 AM” es claramente una de esas expresiones, y su letra llega a aludir a un suicidio. La figura representada en el centro de la escena está enunciada en segunda persona. En la grabación original difundida en La grasa de las capitales, a partir de la mención de la palabra “reloj” en el sexto verso de la primera estrofa, comienza a sonar al tic-tac de un reloj pulsera, que sólo cesa con la onomatopeya “Bang, bang, bang” en la mitad de la última estrofa. El locutor, claramente ubicado en un lugar de saber, describe las actitudes y movimientos del tú y recuerda aspectos de un derrotero de vida atribuido a este último, marcado por diversas búsquedas infructuosas. Uno de los efectos de la peculiar articulación enunciativa es la construcción de un tú “en tela de juicio”, que, sin voz, escucha la interpretación que de su propia persona y de su vida hace otro ser. El yo que se construye en esa voz “evaluadora” comparte con el tú la premisa de la necesidad de búsqueda de un espacio diferente e ilimitado, representado en la inmensidad “de un sol y de un mar” y en el rechazo a la seguridad sugerido por la “vida peligrosa” soñada. Ambos aparecen, de ese modo, relacionados con el universo de valores del campo roquero. Pero el saber escéptico del yo locutor condena la insuficiencia de ese deseo frente a lo que “en sí” no cambiará. Otro efecto de la enunciación construida es que los dos lugares, tanto el del yo como el del tú, están más disponibles para ser ocupados por el virtual alocutario que lo que estarían si se hubiese puesto en escena un personaje en 3ª persona. Se puede ocupar el lugar del desahuciado como escuchas de la canción, pero su entonación e “incorporación”, con el valor que atribuimos a ese término en 2.2, nos desplazan al lugar de ese evaluador casi-juez. FANJUL. Tallado y demarcación... 368 \| Linguagem em (Dis)curso Los “bang” de la última estrofa sacan de escena al tú. Sólo quedan “hojas muertas que caen”. El locutor pasa a referirse a una 3ª persona general y adquiere él mismo un tono proverbial (“los que no pueden más se van”), reforzado por el introductor “siempre igual”. El drama y los contornos atribuidos a esas instancias personales van siendo construidos a lo largo de la letra mediante una sucesión de imágenes que iremos analizando a la par que observamos la versión creada en Brasil muchos años después. Ya en el comienzo, la versión en portugués muestra dos diferencias que producen una representación del cuasi-personaje tú mucho menos autodestructiva. Por un lado, en la letra de Charly García, se atribuye al mismo tú esquivar su corazón y destrozar su propia cabeza. En la versión de Vianna, cierta ambigüedad sintáctica determina que el causador de esos procesos puede ser a febre de um sábado azul e um domingo sem tristezas, y el tú esté en un papel de experimentante4. Por otra parte, la imagen de “destrozar” la cabeza pasa a ser de certezas destruidas. En función de lo primero, en la letra argentina el tú / personaje aparece más centrado sobre sí, su relación con el entorno es de separación. Una huella de esa escasa vinculación es la discontinuidad sintáctica entre las dos frases nominales del inicio (La fiebre de un sábado azul y un domingo sin tristezas) y la cláusula que les sigue. En la versión brasileña se registra una interacción en que elementos del entorno son activos para una alienación del ser en relación con sí mismo. En la letra en español también está afectada en cierta medida la integración entre dimensiones del ser, pero es el propio personaje quien aparece representado manipulando las “partes”. Si “esquivas a tu corazón” aparece como una cierta desatención a sus propios sentimientos, “destrozas tu cabeza”, en cambio, puede verse como una versión negativamente valorizada de la experiencia alucinatoria, el “reviente” como contracara del “viaje”, si no como anticipación “literal” 4 En efecto, podemos interpretar “a febre te esquiva” o, siguiendo una concordancia posible en algunas variedades del portugués brasileño, “(você / tu) te esquiva”. No ignoramos que interviene en esa transformación el régimen pronominal del verbo esquivar(-se) en portugués. Sin embargo, como ya anticipamos en 2.2, esas condiciones no explican el resultado, ya que habría muchas paráfrasis posibles que mantendrían un papel de agente para el tú. Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 369 del suicidio. Y aquí entra lo segundo, destrói tuas certezas, que introduce una dimensión de conocimiento que no estaba en juego, al menos de esa manera, en la letra de García, y refuerza la asimetría entre el tú y el yo en cuanto al saber. El tú interpelado y puesto en escena en la letra argentina es agente de su propia devastación; en ese sentido, aún en su miseria humana, es compacto. Su émulo en la versión brasileña evidencia en su contorno la acción del afuera. El primero parece “decidir” arrasar con su propia mente, pero no le ha pasado que ésta se le escape, como ocurre al segundo. En las estrofas segunda y tercera se expone el conflicto mediante un gesto narrativo en que el tú se acerca más a la condición de personaje. Se lo relaciona con el ámbito de la creación musical y en general, como anticipamos, con valores compartidos en el campo del rock. Pero de entre esos valores, el de la autenticidad parece afectado por algunos de los cambios que se le atribuyen: cambiar de música, de ideas, de color. Son las estrofas que en la versión de Vianna siguen el camino de una traducción casi literal. Algunas excepciones son “gris ciudad” por cinzas 5 y “fronteras”, que en la Argentina de 1979 remitía a los muchos exilios y autoexilios6, por bandeiras. Además, no suena en la versión brasileña, en ningún momento, el reloj de fondo. Pero es en la cuarta estrofa que se produce la transformación más significativa en cuanto a la enunciación, acompañada, como veremos, por algunas otras marcas congruentes. En la letra en español, hasta los bang, se mantiene la estructura enunciativa que ya describimos: un yo que habla a / sobre un tú / personaje: “Y llevas el caño a tu sien…”. En cambio, en la letra en portugués se produce ahí una notable ambigüedad al respecto, posibilitada por el funcionamiento del portugués brasileño, pero que también podría, en esa lengua, haber sido evitada con diferentes procedimientos que diesen a los versos univocidad en la referencia personal. De hecho, hay tres maneras posibles de leer esa referencia: 5 En portugués, la palabra cinzas puede equivaler a, en español, “cenizas”, pero como adjetivo y en singular, cinza, en portugués, es también el nombre del color gris. 6 Varios de los miembros de Seru Giran venían de un autoexilio precisamente en Brasil, donde de hecho empezó a formarse la banda. FANJUL. Tallado y demarcación... 370 \| Linguagem em (Dis)curso a) El yo locutor le habla al tú, informando lo que el tú hace (la misma de la letra en español): [eu vejo que você] levanta o cano outra vez e aperta… b) El yo cambió de perspectiva y muestra al personaje como 3ª persona, informando lo que esa persona hace: [eu vejo que ele] levanta o cano outra vez e aperta… c) El yo le habla al tú indicándole lo que debe hacer: [eu peço / ordeno a você:] levanta o cano outra vez e aperta… Nótese que la posibilidad (a), que corresponde a la de la letra de García, es la menos disponible, en el portugués brasileño, para esa formulación, ya que para producir esa referencia, tendería a retomarse pronominalmente el sujeto: Você levanta o cano outra vez…. Esa ocurrencia de você sería, además, armónica con la estructuración de la tercera estrofa, con la cual guardaría paralelismo. Y en el lugar donde podría estar aparece un então, que permite tanto una lectura temporal (nesse momento), que se acercaría al valor de conector narrativo del “Y…” que encabeza la estrofa en castellano, como una interpretación consecutiva. Creemos que esa estructuración tiene efectos en tres órdenes. Por una parte, su ambigüedad en cuanto al tipo de enunciación (¿narración o apelación?) consolida la disposición de evaluador-juez que ya atribuimos al yo locutor, así como su lugar de saber, reforzado por el posible valor explicativo de então. Por otra parte, debilita más aún lo autodestructivo en la representación del tú, y da más primacía al entorno por descompactar el cierre del ser sobre sí. A ese segundo efecto contribuye también la diferencia entre “ves todo el mar en primavera” y vê um céu de primavera, imagen que, dada la ambigüedad referencial, puede incluso no surgir, como visión, del mismo ser. Y por último, pero no menos importante, desestabiliza el carácter “omnisciente” del locutor-evaluador y crea un nuevo espacio personal. Ese espacio es una modalidad intermedia entre el centro absoluto de escena que el tú / personaje ocupa en las tres primeras estrofas y su total disolución, después de los “bang”, en una persona general. En la letra argentina hay entre ambas situaciones un corte abrupto, y en la versión brasileña aparece ese lugar de transición que es extremamente inestable, ya que oscila entre el de un personaje pleno y el de un interlocutor. Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 371 El conjunto de las transformaciones enunciativas en la versión brasileña apunta a la configuración de un contorno para el tú-personaje menos compactamente delimitado que en la versión argentina. Considerando que el contorno, como todo proceso de determinación, resulta del embate entre lo que se recorta y lo que queda afuera, observamos la versión otra como oportunidad de ver algo de lo residual, constitutivo, del primer tallado. La transposición brasileña realiza sus propias exclusiones pero también recoge lascas que cayeron de la primera. Destacamos la posibilidad latente para el locutor y, por incorporación, también para el interlocutor-oyente, de prever o hasta regir los movimientos del ser en escena. Combinación, en la voz, de la empatía con la víctima y la mirada del victimario. Un goce específico del lugar disidente que en el rock argentino sólo emergerá años después, revestido de un cierto cinismo, en los dúctiles enunciadores escenificados por Los Redondos7. 4 BUSCANDO UN SENTIDO A TODO ESTO Nos ocuparemos ahora de un recorrido en sentido inverso: una composición brasileña reformulada por una banda argentina. “El tiempo no para” es parte de la etapa solista de Cazuza, considerado uno de los más grandes letristas del rock de Brasil, con fuertes lazos poéticos con tradiciones anteriores de la canción urbana. En la letra de ese clásico, el centro de escena es ocupado por un yo que realiza un casi manifiesto sobre su lugar en el mundo. En varios momentos se interpela a un tú, que, sobre todo en la última estrofa, puede verse como generalizador (Te chamam de ladrão, de bicha, maconheiro…). En la misma estrofa aparece también una tercera persona general8 que 7 Patricio Rey y los Redonditos de Ricota. Banda de rock argentina de las décadas del 80 y el 90, de marcadísima singularidad, principal exponente del underground. en el país. El artículo citado de Monteleone se centra en el análisis de su poética. 8 Utilizaremos las denominaciones “persona general” y “generalizador”, empleadas entre otros por Fernández Ramírez (1987), porque rechazamos las de “indeterminación del sujeto” o “impersonalidad” que se encuentran en algunos espacios de la reflexión gramatical sobre esas formas. La primera, porque creemos que es imposible la no determinación en el discurso: inclusive la indefinición y la genericidad son modos de determinación. La segunda, porque desde el punto de vista sintáctico, hay sujeto en todos los casos. FANJUL. Tallado y demarcación... 372 \| Linguagem em (Dis)curso resume la figuración opuesta a la del yo, construida a lo largo de toda la canción a partir de la imagen de correr na direção contrária y otras que le siguen y que presuponen una adversidad activa. Esa persona general opresora es relacionada en determinado momento con los que detentan poder sobre el país al punto de poder transformarlo en un puteiro (prostíbulo). En las condiciones impuestas por ese poder, restricciones que no ofrecen escapatoria (é matar ou morrer), un “nosotros” va conformando su condición: e assim nos tornamos brasileiros. Si bien a partir de la aparición de ese “nosotros” el foco de la letra se desplaza hacia esas formas generalizadoras, la perspectiva dominante es la del yo locutor. Ese ser, como en otras composiciones del género, aparece dotado de visión que otros no tienen y adquiere un tono proverbial en la aserción del estribillo, repetido en el cierre y que también da título al tema: o tempo não pára. La banda argentina Bersuit Vergarabat grabó la composición en su primer LP. Cazuza era, y todavía es, casi un desconocido en Argentina, y Bersuit recién comenzaba a darse a conocer. La versión de O tempo não pára, si bien siempre fue presentada evidenciando su proveniencia (en muchas ejecuciones en vivo el estribillo se ha cantado en portugués) muestra en un punto un intento de reubicación de la escena en un nuevo contexto nacional. En efecto, assim nos tornamos brasileiros fue traducido como “así nos hacemos argentinos”, en lo que más parece una adopción que un cambio: identificar los mecanismos de opresión y corrupción como semejantes a través de las fronteras nacionales. Dejando de lado ese detalle, podemos decir que la adaptación argentina muestra un notable trabajo de adhesión a la propuesta de una composición de partida, que contiene momentos de una elaboración poética nada simple. Sin embargo, vemos que en esa adaptación se produce un conjunto de desplazamientos por medio de los cuales el locutor realiza un trabajo que denominaremos sobredemarcación. La letra de Cazuza expone de manera contundente un conflicto, incluso representable espacialmente (correr na direção contrária), entre la perspectiva en que el yo se incluye y la del poder que aparece como fuerza opuesta. Intentaremos mostrar en nuestro análisis que si la separación entre el espacio de una fuerza y de otra puede verse, en Cazuza, como un trazado discontinuo pero recuperable, en Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 373 la versión de Bersuit es como si el mismo se remarcase con una línea ostensiva que distribuye todas las entidades personales inequívocamente de un lado o del otro. Ese trabajo va acompañado por un cierre, realizado sobre todo mediante el agregado de conectores, de relaciones semánticas que en la letra de Cazuza se encuentran menos definidas. Desarrollaremos nuestra explicación siguiendo la progresión de la letra. En el comienzo, la imagen del yo disparando contra el sol es seguida, en la letra de Cazuza, por la formulación sou forte, sou por acaso. En “disparo contra el sol con la fuerza del ocaso”, si la fuerza atribuida al yo se mantiene o hasta se realza, ya que el ocaso es la desaparición del sol, se elimina, en cambio, la tensión entre forte y por acaso, expresión esta última que remite a lo casual, a lo no motivado. En la misma línea, vemos la transposición de mágoa (rencor) por “magia” como parte de la construcción de una imagen de fortaleza. Creemos que aunque la magia no es mostrada como una herramienta eficaz, no deja ser un atributo que confiere algo de especial al ser que se construye en ese inicio de la versión en castellano, dibujado sin la herida de ser por acaso. Así, el cambio en el modo de conexión con lo que sigue parece hacerse necesario. Donde simplemente teníamos la afirmación. Eu sou um cara, sin conector, tenemos una formulación fuertemente contrastiva, “pero sólo soy un hombre más”. La restricción “sólo uno más” va en el sentido de la reformulación, en la letra de Cazuza, unos versos después: Eu sou mais um cara. Pero la construcción adversativa es realmente un agregado de la versión argentina: la fuerza y la magia no son suficientes para el combate. Con la misma linealidad con que se construyó el personaje, se deja “constancia” del alcance de su fuerza y se clausuran otras relaciones posibles, de no oposición, entre la imagen inicial y ser um cara 9. Luego de la imagen ya mencionada de correr en dirección contraria, tenemos: sem pódio de chegada ou beijo de namorada. Eu sou mais um cara sin podio de llegada y mi amor me corta la cara, porque soy sólo un hombre más. 9 Relaciones causales, por ejemplo: minha metralhadora cheia de mágoas [porque] eu sou um cara cansado de correr na direção contrária. FANJUL. Tallado y demarcación... 374 \| Linguagem em (Dis)curso La simple inexistencia (genérica) de una novia se trasforma en el rechazo por parte de una pareja específica. Y el hecho de ser “sólo un hombre más” pasa a ser causa de ese rechazo. El amor abandonador, que esperaba algo más que sólo un hombre, es puesto fuera del espacio del yo. Enseguida, se introduce una nueva relación semántica, causal, que cierra la interpretación. Por ser sólo un hombre, la falta de gloria (podio de llegada) lleva al abandono, a pesar de tratarse de un ser dotado de fuerza y potencial magia. En la estrofa siguiente, conectada, en ambas letras, como adversativa en relación con la anterior, se relativiza la disparidad de fuerzas: el yopersonaje niega estar derrotado, pensamiento atribuido a un tú que no parece identificable como otro personaje, sino un modo de, entre las diferentes formas de generalización (se alguém pensar, se pensarem, si se piensa, si piensan), apuntar al virtual interlocutor de la proclama. La réplica es saiba que ainda estão rolando os dados, vertida como “quiero que sepas que me la sigo jugando”. Comparando las dos construcciones empleadas, resulta bastante diferente el involucramiento representado para el ser que toma voz en el yo. El juego continua, y en la aserción en portugués están dados, por contexto, la participación y el riesgo. Pero éste está mucho más sujeto al devenir externo que en “me la sigo jugando”, expresión de arrojo que involucra una dimensión mucho más íntima y tendiente a la representación de un individuo: el ser está en el centro del juego, es a la vez su disparador y su objeto. Algunos versos después, en la imagen de la piscina, se interpela a otro tú, este mucho más singularizable: A tua piscina está cheia de ratos, tuas idéias não correspondem aos fatos. Y tu cabeza está llena de ratas, te compraste las acciones de esta farsa. En la versión argentina es más nítida la expulsión del tú hacia el campo de lo que el yo–personaje aparece combatiendo desde el inicio de la letra. La modulación de Cazuza en el canto es verdaderamente asqueada, pero en su letra las ratas se representan en el ámbito de las posesiones del ser interpelado, no en el de su personalidad (“cabeza”). Además, lo que puede verse como una equivocación (tuas idéias não correspondem aos Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 375 fatos) aparece como adhesión a un engaño deliberado (“farsa”), el error se transforma en estafa. El siguiente desplazamiento que destacamos, en la estrofa posterior, se refiere a cómo “emplea sus días” la figura central: procurando uma agulha no palheiro / “buscando un sentido a todo esto”. La diferencia va en el mismo camino de construir un yo-personaje estabilizado en relación con el entorno. Análogamente a lo que observábamos en el caso de “Viernes 3 AM” para la disposición del ser en escena, en la letra brasileña, buscar una aguja en un pajar parece mostrar una tarea infructuosa por parte de un ser desorientado. En la versión argentina, se observa una disposición de propósito, un impulso de control sobre el entorno, que puede resultar igualmente inútil, pero muestra el espacio del yo–personaje como racionalizado. Incluso, como huella de la transposición de la letra a la discursividad de otro espacio lingüístico-nacional, “buscando un sentido a todo esto” puede verse como meta-enunciación que describe, impensadamente, la tarea desplegada por el enunciador-reformulador: el intento de control de las posibilidades semánticas, insertando en la letra numerosas señales que orienten su interpretación. En la última estrofa antes de la repetición del estribillo, un nuevo tú generalizador es reemplazado por un “nosotros”: Te chamam de ladrão, de bicha, maconheiro / “Nos tildan de ladrones, maricas, faloperos”. La perspectiva es empática, se le habla a quien comparte el mismo lugar de oprimido. En ese sentido, la segunda persona, en la letra de Cazuza, muestra movilidad respecto de la orientación argumentativa de la voz instalada como yo, ya que en las dos apariciones anteriores la relación es de oposición polémica. No ocurre lo mismo en la versión de Bersuit, donde se produjo una separación, como vimos, mucho más tajante en relación con el tú. Así, el “nosotros”, que ya apareció en “así nos hacemos argentinos” y está indisolublemente ligado al yo-personaje, es convocado hacia el lugar generalizador. Por último, uniendo esa afirmación con la que sigue (Transformam um país inteiro num puteiro…), encontramos en la letra argentina un agregado que explicita el contraste deducible en el original: “y ellos sumergieron un país entero…”. De ese modo, los espacios opuestos quedan sobredemarcados, y cada ser representado ubicado “en su lugar” en relación con el personaje central, en este caso, aparentemente, no sujeto a nada que, en el plano del enunciado, desdibuje su contorno. FANJUL. Tallado y demarcación... 376 \| Linguagem em (Dis)curso 5 CONCLUSIONES Las composiciones de las que nos hemos ocupado recogen en su dialogismo, de diferentes maneras, polémicas en el campo del rock, en toda su extensión mundial, sobre la necesidad de transformar la sociedad y la vida cotidiana, y la posibilidad / imposibilidad de hacerlo, así como sobre el alcance, individual o colectivo, de esas transformaciones. Ni “Viernes 3 AM” ni O tempo não pára se encuadran en el rock de visión revolucionaria ni en sus desdoblamientos utopistas: a pesar de la diferencia de 9 años entre sí, ambas son claramente pos-hippismo y pos-Vietnam, y en su materialidad resuena la desazón generacional que se expresó mediante el punk y tantas otras manifestaciones de fines de los 70’ y comienzos de los 80’. Analizamos a lo largo de este artículo, en las dos letras y sus versiones, la construcción y el tallado de dos seres, uno de ellos en segunda persona, construido por una voz, y otro en primera persona, construido por y en la voz. Ambos representados en un entorno de adversidad en el que alcanzan diferentes grados de heroicidad. Y aunque en ambos se advierten aspectos del bagaje ideológico ligado al decir roquero, en el primer caso la relación del locutor con el ser es por momentos polémica, y en el segundo, de plena confluencia en una misma voz y posicionamiento. Nos propusimos observar, en sus transposiciones a la otra lengua y a la otra discursividad, desplazamientos en los contornos trazados para esas entidades personales. Si podemos identificar un movimiento relativamente repetido, fue el de integración del ser sobre sí en la producción del rock argentino y de permeabilidad con el entorno en las letras brasileñas. Así, en “Viernes 3 AM” se representa la derrota de un sueño de trasformación, y la autodestrucción es la expresión de esa derrota, levemente desplazada, en la versión brasileña, a una desestructuración del ser en el entorno. En O tempo não pára, la derrota se refuta en la voz de un ser que resiste la adversidad desde un lugar de preeminencia, pero que no excluye la desorientación y el reconocimiento de un devenir (o acaso, o rolar dos dados), que se le escapa. En la lectura desde Argentina, la refutación de la derrota se desliza hacia una proclamación de la perspectiva personal (“me la sigo jugando”) y de su intento de control sobre el entorno hostil. Linguagem em (Dis)curso, Tubarão, SC, v. 13, n. 2, p. 359-381, maio/ago 2013. Linguagem em (Dis)curso \| 377 No es el optimismo o el pesimismo lo que las diferencia, como podría pensarse a partir de conocidos estereotipos. Los rock (y, en general, la música popular) de ambos países han cantado victorias, derrotas, esperanza y descrédito en el diálogo polémico del campo del que forman parte en el mundo. En ambos se han diseñado héroes exultantes y melancólicos, desgarrados y apacibles. Lo que parece marcar una tendencia diferenciadora es la expectativa para su tallado, el juego que otorga máscaras a la voz disidente en posicionamientos análogos de resistencia. REFERENCIAS BENVENISTE, É. Problemas de lingüística general. Vol. 2. México: Siglo XXI, 1971.	38,641
https://openalex.org/W4293076471	OpenAlex	Open Science	CC-By	2,022	1 Einleitung	null	German	Spoken	1,106	2,314	Open Access. © 2022 Joachim Gerdes, publiziert von De Gruyter. Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz. https://doi.org/10.1515/9783110770193-001 1 Einleitung Die Fachsprachenforschung ist nicht nur in der germanistischen Linguistik eine mittlerweile weit gefächerte und relevante Disziplin. Es gibt zahllose wis- senschaftliche Monographien und Artikel zu verschiedensten Themenstellun- gen dieses Forschungsbereiches. Insbesondere im Hinblick auf die Natur- und Technikwissenschaften spielen Erkenntnisse zu fachsprachlichen Strukturen und Terminologie, deren letztere mittlerweile einen eigenen Wissenschaftszweig darstellt, eine wesentliche Rolle für die Optimierung der jeweiligen Fachkommu- nikationen, nicht zuletzt auch im Bereich der Übersetzungs- und Dolmetschwis- senschaften. Auch im Bereich der Geisteswissenschaften ist die Analyse der Fachsprachen ein relevantes Forschungsgebiet, das sich zunehmender Aufmerk- samkeit erfreut. Die protestantische Theologie als institutionalisierte Universitätswissenschaft gehört keinem der Bereiche eindeutig an, lässt sich aber aufgrund ihrer Inhalte und Forschungsmethoden am ehesten den Humanwissenschaften im weitesten Sinne zuordnen, wo Grenzwissenschaften zu ihren Subdisziplinen angesiedelt sind, wie etwa die Geschichtswissenschaft, die mit der Kirchengeschichte eng ver- knüpft ist. Ebenso sind die exegetischen Wissenschaften (Altes und Neues Testa- ment) mit den altsprachlichen Philologien verschwistert, und schließlich ist die systematische Theologie unmittelbar mit der Philosophie verbunden. Zur Fach- sprache und wissenschaftlichen Terminologie der theologischen Disziplinen, ins- besondere der Systematik und Dogmatik, liegen jedoch, abgesehen von einzelnen Artikeln, nur äußerst wenige wissenschaftliche Studien vor. Diesem Mangel soll mit dem vorliegenden Buch begegnet werden, indem ein gründlicher Überblick über Haupttendenzen der protestantisch-theologischen Fachsprache vom Ende des 19. Jahrhunderts bis zum Beginn des 21. Jahrhunderts präsentiert wird. Das Phänomen einer über eine sachlich-neutrale Darstellung des Inhaltli- chen hinausgehenden sprachgestalterischen Komponente spielt für die theolo- gische Fachsprache eine entscheidende Rolle. Daher drängt sich die Frage auf, ob diese durch ihre spezifische Gestalt als eine Art Hybridsprache aus appellati- ven, deklarativen und auch spekulativen Elementen besonders anfällig für ideologische Entstellung und inhaltliche Vagheit ist und damit die in politisch- historischen Kontexten variierenden Konnotationen der Fachbegriffe leichter der Vereinnahmung durch außerfachliche Diskurse anheimfallen lässt. Dies soll anhand eines historischen Überblicks über sprachliche Besonderheiten bei repräsentativen theologischen Autoren und in vorherrschenden wissenschaftli- chen Strömungen seit dem Ende des 19. Jahrhunderts untersucht werden. Es gilt, die Hypothese zu überprüfen, ob es nicht zuletzt auch sprachliche, d. h. 2 1 Einleitung 2 2 lexikalische, semantische, stilistische und pragmatische Operationen sind, die bewusst oder unbewusst zu neuen Sichtweisen auf vermeintlich unumstößliche und über Jahrtausende überlieferte religiöse Botschaften führen. 1 Einleitung Zur Methodik sei angemerkt, dass es sich als ergiebig und zielführend erwiesen hat, die jeweili- gen epochenspezifischen Charakteristika der theologischen Fachsprache anhand besonders bedeutender, Schule bildender oder zeittypischer Wissenschaftsper- sönlichkeiten zu exemplifizieren und zu veranschaulichen und auf dieser Grund- lage Schlussfolgerungen im Hinblick auf allgemein vorherrschende Tendenzen zu ziehen. In Kapitel 2 bildet zunächst eine Übersicht über Merkmale der protestan- tisch-theologischen Fachsprache im Allgemeinen und ihre Darstellung in der Fachsprachenforschung den Auftakt. Die acht Hauptkapitel (3‒10) befassen sich mit der Fachsprache einschlägiger, für die jeweilige Epoche prägender Theologen und theologischer Schulen. Als prägende Gestalt der protestantischen Theologie der wilhelminischen Epo- che und des ausgehenden Kaiserreiches wird zunächst in Kapitel 3 die Sprache des von Maximilian Harden als „Hofdogmenlehrer“ bezeichneten Adolf von Har- nack untersucht (vgl. Bruch 2006: 89), der neben weiteren Theologen wie Adolf Schlatter oder Gustav Adolf Deissmann insofern nicht zufällig im Mittelpunkt des Interesses steht, als sein Schüler Karl Barth als Hauptrepräsentant der ‚Theologie der Krisis‘ sich später auch sprachlich radikal von Ersterem loszusagen bestrebt ist. Ausgehend von der liberalkonservativen Theologie des späten 19. Jahrhunderts und ihrer Sprache der bürgerlichen Selbstgenügsamkeit wird in Kapitel 4 ein Bogen zu eben dieser auch in linguistischer Hinsicht einschneidenden, durch den Schweizer Theologen Karl Barth ausgelösten Wende der ‚Theologie der Krisis‘ ge- schlagen. Neben Barths Sprachrevolution steht im Mittelpunkt des Kapitels ferner die Fachsprache weiterer Protagonisten der ‚Dialektischen Theologie‘ wie Emil Brunner und Eduard Thurneysen. Anhand signifikanter Textauszüge soll im Anschluss daran in Kapitel 5 die Sprachentwicklung führender Theologen, die während der NS-Herrschaft forschten und lehrten, beleuchtet werden, darunter Gerhard Kittel, Paul Althaus, Emanuel Hirsch, Friedrich Gogarten und andere. Im Kontext der Totalisierung, Ideologisierung und Brutalisierung der Sprache durch die nationalsozialistische Propaganda und Sprachlenkung stellt zweifellos die protestantisch-theologische Fachsprache einen besonderen Fall dar. Im Bereich der akademisch-theologischen Wissenschaftssprache erscheint eine linguistische Analyse im Hinblick auf deren Aufnahmebereitschaft gegenüber der nationalsozialistischen Ideologie- sprache aufschlussreich, zumal umgekehrt die Sprache der Religion und der Theologie in ihren unterschiedlichen theoretischen oder kirchenpraktischen Erscheinungsformen per se einen dogmatisch-missionarischen oder auch au- 1 Einleitung 3 1 Einleitung 3 3 toritativ-propagandistischen Charakter hat und somit in der Wahl der sprach- lichen Mittel immer wieder als Vorbild für den Jargon totalitärer politischer Ideologien dient. 1 Einleitung Wenn die christlich-protestantische Verkündigung jedoch auf Werten wie Toleranz, Nächstenliebe, Brüderlichkeit, Menschlichkeit und dem Primat eines individuellen Gewissens beruht, drängt sich die Frage auf, wie sich der sprachliche Ausdruck solcher essenzieller Wertvorstellungen mit dem Vokabular der Unterordnung, der bedingungslosen Gefolgschaft und des Gehorsams vereinbaren lässt und wie sich die protestantische Predigtsprache und die theologische Fachsprache allgemein im Kontext autoritärer politisch- gesellschaftlicher Systeme und deren invasiven Sprachpolitik entwickeln. Kapitel 6 widmet sich dagegen der Fachsprache des theologischen Wider- stands zwischen 1933 und 1945; hier stehen insbesondere die Stimmen von Dietrich Bonhoeffer und Martin Niemöller im Vordergrund. Im darauffolgenden Kapitel 7 werden sprachliche Kontinuitäten und Neuorientierungen in der Nachkriegstheologie in der Bundesrepublik Deutschland betrachtet, sowohl im Hinblick auf Publikationen bereits vor 1945 aktiver Theologen als auch auf Schriften jüngerer Theologen wie Gerhard Ebeling und Helmut Thielicke. Kapitel 8 widmet sich der theologischen Fachsprache in der Bundesrepublik des Nachkriegswiederaufbaus und in der Zeit der sich in der protestantischen Theologie besonders vehement manifestierenden progressiv-reformistischen Ten- denzen der 68er-Bewegung, in deren Umfeld auch die wissenschaftliche Theologie in den Sog einer gesellschaftspolitisch engagierten Aufbruchstimmung und eines auch in der Fachsprache sich manifestierenden Traditionsbruches gerät. Hier ste- hen u. a. die Namen Dorothee Sölle, Helmut Gollwitzer und Wolfhart Pannenberg im Mittelpunkt. Kapitel 9 wirft ein Licht auf die protestantisch-theologische Fach- sprache in der offiziell atheistischen, kirchen- und religionsfeindlichen DDR, die die universitäre Theologie zu einem weitgehenden Nischendasein unter ideologi- schem Anpassungsdruck zwang. Das Schlusskapitel 10 widmet sich aktuellen Ent- wicklungen in der theologischen Fachsprache im wiedervereinigten Deutschland der 90er Jahre und des beginnenden 21. Jahrhunderts und beleuchtet Neuausrichtungen der protestantischen theologischen Wissenschaftslandschaft in einem gesamtdeut- schen, politisch-ideologisch weniger akzentuierten, administrativ und personell pro- fessionell durchorganisierten akademischen Lehr- und Publikationsbetrieb. Es handelt sich bei der vorliegenden Studie ausdrücklich um keine theolo- gische Abhandlung, sondern um eine sprachwissenschaftliche Betrachtung der Fachsprache(n) der protestantischen, vorwiegend lutherischen, aber auch re- formierten Theologie. Da Sprache und Inhalt nicht immer klar voneinander zu trennen sind, bleibt es nicht aus, dass hier und da Reflexionen zu theologi- schen Problemstellungen zur Sprache kommen, die jedoch grundsätzlich im Dienst einer linguistischen Fachsprachenanalyse stehen.	2,760
tel-04624661-2023_ERROUANE_arch.txt_24	French-Science-Pile	Open Science	Various open science	null	Paratraduction, Transcréation et communication : pour la promotion du Tourisme et la sauvegarde du Patrimoine en Algérie. Sciences de l'Homme et Société. Université Paul Valéry - Montpellier III, 2023. Français. ⟨NNT : 2023MON30050⟩. ⟨tel-04624661⟩	None	French	Spoken	6,820	11,537	Émilie Flon dans « mise en scène du patrimoine, savoir, fiction et médiation » explique que des outils comme l’iconographie, panneaux, fresques, ou autres objets exposés dans un site historique peuvent être considérés comme des éléments de médiation pour la mise en scène de ce site laquelle permet de : 1. Le faire exprimer, en ressuscitant son passé à travers l’utilisation de ses fragments (comme le cas des ruines par exemple) d’un contexte antérieur à un autre présent et sous de nouvelles circonstances ; 2. Le faire exprimer dans l’objectif de raviver une mémoire collective et dans ce cas, toute son histoire devrait être reconstruite par les spectateurs/visiteurs ; 3. Émouvoir et susciter des réactions chez les spectateurs car les traces du passé ne sont pas autant interrogées pour se rapprocher la réalité historique que pour partager et transmettre des valeurs (cf. Flon, 2012, pp. 16-17). À propos de la mémoire collective, le géographe Stéphane Héritier estime que « les lieux touristiques servent à amarrer la mémoire tout en ouvrant, dans le même temps, la possibilité de la (re)configurer. La mémoire associée à un lieu est évolutive et ce d’autant plus qu’elle se recharge de signification à mesure de la transformation des sociétés et du regard que ces dernières portent sur leur passé » (Héritier, 2018, p. 37) et « la fréquentation des lieux patrimoniaux, désignés, promus et commémorés permet de favoriser en retour dans une sorte de mécanisme itératif, leur reconnaissance en tant que composante de la mémoire collective et nationale » (ibid. p. 39). En reprenant l’expression de « mise en scène du patrimoine » Daniel Jacobi et Maryline le Roy estiment que les monuments, lorsqu’ils sont en phase promotionnelle, deviennent des équipements et des offres culturelles (cf. Jacobi et le Roy, 2013, p. 19). Ils sont mis en valeur par la signalétique à laquelle, les auteurs joignent les panneaux et les étiquettes en tant que médias d’exposition « endotextuels » qui diffèrent des autres écrits habituels (plans, dépliants, livrets, guides de visite) dits « exotextuels » ; les auteurs rajoutent ensuite dans ce qu’ils ). appellent « registres d’ à l’interprétation », les textes multimédias. La signalétique est une forme textuelle singulière parce qu’elle porte un code scriptovisuel original pouvant être accompagné de signes de valeurs symboliques et du contenu textuel culturel. Les fonctions du balisage et du jalonnement permettent de pointer et de nommer les aspects saillants d’un parcours, elles satisfont les besoins d’orientation spatiale et instaurent une régulation du parcours de visite. La fonction textuelle permet d’identifier, de désigner, de fournir des renseignements sur les noms des espaces et d’indexer ce qui est pertinent. (Ibid., pp. 18-19). Dans le même ordre d’idées, le texte et le discours (et l’on peut même, pour notre part, inscrire la signalétique) sont aux yeux de Stéphane Héritier des composantes du dispositif de narration qui caractérisent les récits en tant qu’instruments de production des lieux puissants et nécessaires ; la narration permet de réveiller des histoires endormies et « narrativiser » est une action qui se traduit par une mise en cohérence d’éléments spatialement épars sous la forme d’un récit, c’est ainsi que le discours permet d’expliciter au mieux l’histoire d’un lieu touristique et sa configuration sociale, il est le processus de médiation qui se déroule en trois étapes : a) l’intention : c’est-à-dire l’intention de communiquer ; b) la formalisation : laquelle vise à inclure le dispositif spatial dans des supports) et ; c) la présentation : laquelle est mise à disposition du public (cf. Héritier, 2018, p. 43). Si on applique cette physionomie aux sites que nous avons visités durant notre enquête, on réalise qu’ils ne sont pas tellement concernés par les modes de reconfiguration spatiale (restauration), ni par les dispositifs de mise en scène ou de médiation que nous avons mentionnés plus haut, on obtiendra donc des bâtiments condamnés, métaphoriquement parlant, au silence absolu, isolés de tout processus d’animation compte tenu de l’absence de dispositifs technologiques permettant de les transformer en offres culturelles, de renforcer leur authenticité et de les rendre plus attractifs. Par conséquent, cela entraîne non seulement une rupture de transmission de valeurs historiques, mais aussi une rupture de la communication patrimoniale intergénérationnelle que malheureusement les quelques panneaux rudimentaires disposés à quelques endroits sont insuffisants pour pérenniser leur prolongement dans la mémoire. 2.2. Transformation numérique et multimodalité La transformation numérique constitue le passage d’un monde à l’autre, du réel au virtuel. Dans cette phase, l’espace touristique peut faire l’objet de traitements infinis permettant de renouveler son attractivité, lui attribuer d’autres valeurs par le développement web sans contraintes physiques, matérielles ou spatiales. Plus explicitement, les monuments et sites historiques, après être passés par les phases d’embellissement, d’aménagements divers, de mise en scène et d’interprétation lesquelles se déroulent dans le réel sont « transposés » dans le 378 QUATRIEME PARTIE Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). virtuel pour revêtir encore d’autres procédés d’esthétisation dans les plateformes touristiques. Le but de la transformation numérique est de construire des imaginaires autour d’un site touristique et c’est bien l’imaginaire en tant « qu’espace mental intermédiaire », « phénomène transitionnel » se situant entre deux pôles : « l’espace physique » et « l’espace technologique » qui stimule son attractivité (cf. Amirou, Pauget, et Lenglet, 2011, pp. 89-90-92) : L’imaginaire organise l’espace en parties propices et en parties neutres. Une véritable mise en scène de la vie et de l’espace touristiques se trouve ainsi opérée par les industriels du voyage, mais également par chaque voyageur, à travers les images qu’il emporte avec lui, celles qu’il collecte pendant le voyage, et celles qu’il rapporte chez lui (Amirou, Pauget, et Lenglet, 2011, p. 92). La multimodalité est la caractéristique la plus marquante de la transformation numérique, tout ce qui a trait à l’espace touristique physique est reconstruit dans une page web (ou éventuellement dans une plateforme) par le procédé d’écriture hypertextuelle multimodale : « L’écriture hypertextuelle multimodale est un processus visant à produire un hypermédia, soit une interface textuelle et/ou visuelle constituée d’hyperliens menant vers des documents de différentes natures : vidéo, image, texte. Elle implique l’élaboration d’une architecture textuelle (design/framing) non linéaire, par strate, qui pense en amont l’articulation des modes d’expression et des contenus (complément, redondance, relais, jonction, détournement), leur ancrage (degrés de prédominance), l’effet des choix esthétiques du support de diffusion et de la réception. Ainsi le concepteur/producteur d’hypermédias doit prévoir et consigner les parcours possibles de lecture, l’articulation entre les modes sémiotiques et les usages techniques à l’intérieur du « mode d’emploi » de sa production. De plus, l’hypermédia, façonné par les possibles du numérique devient un mélange interactif d’écrits, d’images, de sons, de graphismes, et participe au renouvellement de l’esthétisme : « le numérique se définit par une diffraction des contenus » provoquant une esthétique particulière, multiple et éclatée » (Audet et Brousseau, 2011, p.10). » (cf. Lebrun & Lacelle, 2015, p. 22). Toutes les informations relatives à un site historique, un musée, ou un parc naturel sont dispersées dans ladite architecture textuelle selon la stratégie de communication préconisée par les concepteurs. Les fragments de textes sont intégrés dans des rubriques auxquelles on associe des symboles, des images fixes et en mouvement (vidéos) et du contenu interactif (des neurs) permettant à un visiteur de faire partie de l’expérience numérique puisqu’il est invité à effectuer un achat, émettre des avis, réserver un parcours de visite, envoyer des doléances, se rediriger vers la communauté web pour prolonger davantage son expérience. Si l’on revient à notre schéma, on s’aperçoit que la transformation d’un site touristique se déroule de manière ordonnée et logique puisque l’on a au début, un site isolé qui sera aménagé, médiatisé ou transformé en équipement culturel...mais avec internet, la découverte des pays Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). ou des métropoles touristiques et leurs équipements culturels se déroule d’abord en ligne, il y a donc presque une « injonction » d’axer les critères performance d’abord sur le volet virtuel qui conditionnerait même pour les plus « méticuleux », le choix d’une destination sur une autre. Avec internet, toutes les régions du monde affichent leurs performances touristiques en offres numériques, parfois même avec exagération, le contenu doit être irréprochable. L’expérience de visite d’une plateforme ou d’un site web touristique se vit à l’intérieur d’un contenu hyperréel, créé à la base d’un univers thématisé où la réalité est recréée et qui peut convenir davantage aux attentes des visiteurs que « la vraie réalité » (cf.Batat & Frochot, 2014, p. 101). L’avantage de l’hyperréalité, en plus de susciter l’engouement des consommateurs à vivre une expérience touristique unique dans un thème particulier, est d’éliminer quelques inconvénients de la destination, d’alimenter le fantasme et la construction – et reconstruction – de l’imaginaire pour présenter enfin un produit culturel dénué de la moindre trace négative. Techniquement, cela se passe sur une plateforme ou sur un site doté d’une interface sur laquelle sont distribués, des images (fixe ou en mouvement) des textes et des sons et des symboles soulignant des labels culturels ou commerciaux. Ces ressources sont reliées entre elles d’une manière compacte, les plateformes présentent généralement comme plan d’accroche des slogans brefs mais puissants avec une rhétorique adaptée au thème du produit touristique vanté, derrière le slogan une séquence vidéo (généralement courte) démarre simultanément lorsqu’on accède au site, elle présente les atouts essentiels du produit (pour un pays touristique, on montrera ses villes attractives, ses monuments, ses parcs naturels, ses touristes semblant témoigner de leur émerveillement et qui prennent ainsi le rôle d’ambassadeurs du pays visité, sa population accueillante, ses officiels, ses prestataires touristiques...). On peut qualifier cette conception homogénéisante des produits touristiques comme un véritable décor, ou scène d’énonciation où plusieurs voies mettent le visiteur au centre de l’action. Pour cette phase le site de tlemcen.mta.gov.dz n’est que peu représentatif des attributions touristiques de la ville au regard des physionomies mentionnées plus haut. Un survol rapide nous permet de comprendre que le site ne présente aucune construction d’hyperréalité ni aucune orientation claire par rapport à un type de tourisme devant être promu (pour Tlemcen, il peut s’agir surtout du tourisme culturel vu le poids historique de Tlemcen et ses monuments). Sur le plan technique, on ne remarque aucune interaction verbo-iconique, aucune vidéo, aucun symbole ou un label et aucun plan interactif pouvant faire adhérer un éventuel visiteur à l’expérience de visite virtuelle, son contenu linguistique, en plus du fait de présenter beaucoup d’erreurs de transposition est orienté dans le discours administratif, ses rubriques relatives à la 380 QUATRIEME PARTIE Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). découverte touristique ne sont pas enrichies avec des textes à vocation publicitaire et certaines d’entre elles n’ont pas de texte. 2.2.3.5 Traduction et paratraduction Tout au long du schéma que nous avons élaboré autour de la critique de notre enquête de terrain, nous n’avons pas fait la moindre allusion à la traduction ou la paratraduction. Nous justifions cela par le fait que l’acte de traduction ou de paratraduction ne peuvent être effectués qu’à la base d’un plan de communication établi adapté à un produit touristique fini. Par conséquent, au vu de la faible performance sur le plan virtuel du site tlemcen.mta.gov.dz et du manque d’interaction verbo-iconique, la paratraduction, pour les objectifs qu’elle suscite – c’est -à-dire associer le paratexte et l’image comme « péritexte iconique », ainsi que d’autres facteurs comme les symboles et les vidéos (sous-titrage) dans le processus de traduction – est irréalisable. En effet, si on applique le schéma de performance touristique sur notre parcours de visite, on réalise qu’à chaque étape, des erreurs sont compromettantes en matière de communication touristique et ce en partant du début, de la prise en charge même de l’espace dont le potentiel naturel peut être exploité aux fins touristiques jusqu’à l’offre sur plateforme numérique. L’accumulation des lacunes à chaque étape que ce soit au niveau de l’aménagement, de la mise en offre touristique, de la mise en scène du patrimoine et sa médiatisation la transformation numérique, ne permet nullement d’exceller en performance touristique par faute de matière et de contenu. Un traducteur ou un paratraducteur, en tant qu’agents communicateurs ne peuvent pas partir de rien. Ainsi pour qu’une bonne traduction touristique soit réalisée, la mise en place d’un contenu riche s’impose dans l’espace physique avec des écrits endotextuels et exotextuels et pour qu’une bonne paratraduction soit effectuée, il nous faut une plateforme touristique bien fournie en ressources sémiotiques verbales et non verbales. 2.2.4 Analyse contrastive des plateformes « algeriatours.dz » et « visitmorocco.com » Avant de nous atteler à l’analyse contrastive des plateformes touristiques algérienne et marocaine, il convient d’apporter quelques précisions à propos de la localisation, la transcréation et la paratraduction. L’acquisition de l’information à l’ère numérique débute sur internet en amont de l’expérience réelle laquelle s’effectue par le déplacement physique. Cela se passe donc à travers les plateformes numériques qui permettent l’accès aux informations instantanées peu importe le centre d’intérêt recherché. Pour le tourisme, le mot « plateforme » a fait l’objet d’un glissement sémantique dans la mesure où il a été amputé de son acception originelle « étendue de terrain relativement plane, située en hauteur par rapport à son environnement » pour revêtir une nouvelle acception décrivant une nouvelle réalité : espace virtuel, informatique permettant l’accès aux voyageurs. La plateforme de promotion du tourisme ou de voyage représente le nœud, le lieu unique par lequel transite l’ensemble des informations ou des services touristiques. Par extension, elle est de surcroît le lieu de transactions où se rencontrent l’offre et la demande de façon dématérialisées au sein d’un écosystème complet de services d’accès intégrés (CNNum, 2014, p. 55). Selon Thomas Jammet et Audrey Linder, la promotion du tourisme par le numérique consiste à composer le voyage de façon virtuelle. Le virage numérique des offres touristiques a fait émerger de nouvelles pratiques sociales liées au tourisme. Ainsi, une plateforme numérique est une fenêtre qui permet de vivre, planifier, réserver et de se mettre d’abord dans une situation de pré-expérience face aux offres touristiques, puis, à travers la personnalisation des choix, le consommateur prend la décision d’aller découvrir le pays pour de vrai. Cette physionomie émane du marketing expérientiel qui stimule le sens du consommateur pour l’attirer dans l’univers de marque afin de lui offrir une expérience gratifiante. Le marketing expérientiel repose sur l’idée que l’individu contemporain cherche à « vivre des immersions dans des expériences de pré-consommation avant de passer à la transaction menant à la consommation effective » (cités par Jammet & Linder, 2019, pp. 1,2 et 3). En conséquence, une plateforme touristique peut-elle faire l’objet d’une paratraduction? dispose-t-elle de toutes les conditions requises à cet effet, c’est-à-dire, un/des paratexte(s), une/des image(s) servant comme péritexte(s) iconique(s), un/des symbole(s) ; ces ressources proposent-elles une lecture combinatoire multisémiotique pouvant renvoyer le visiteur à un contenu hyperréel à partir duquel, il construit son imaginaire? Ces interrogations sont primordiales car l’interaction des diverses ressources internes de n’importe quel site internet en tant qu’espace virtuel de circulation d’information sont supposés influer sur sa réceptivité et sur sa consommation (navigation ordinaire, achat, activité ludique etc...Par conséquent, elles conditionnent son succès ou son échec sur le web. Ce qui lie la traduction aux « plateformes numériques » est constaté, au moins, dans deux activités qui sont : la localisation et la transcréation. La localisation fait référence au processus de traduction et d’adaptation de produits, de contenus et de services aux exigences d’un marché donné. Concrètement, si une entreprise multinationale décide de s’implanter dans un marché situé dans une aire géographique spécifique et aux caractéristiques culturelles différentes (ex. Un groupe industriel européen œuvrant en Asie), tous ses plans marketing, ses supports de communication devront être ( s). élaborés selon une réalité différente, le processus conduisant à la création d’un produit adapté à ladite aire géographique ne peut s’appuyer que sur le volet linguistique (notamment terminologique où il est question ’ , dans le cas d’une adaptation d’un logiciel de comptabilité, de la bonne terminologie d’arrivée), ainsi la localisation implique une combinaison d’aspects linguistiques et technologiques (cf. Scarpa, 2010, p. 339). Pour Jean Delisle, la localisation porte certes sur la traduction d’un contenu verbal selon les conventions textuelle et linguistique de la langue cible, mais la diversité des supports exige que l’on adopte aussi le contenu non-verbal représenté par les couleurs, icônes, graphiques, devises, tailles du papier, unités de mesure, emballage et autres contraintes formelles. De surcroît, le traducteur-localisateur est amené à maîtriser aussi bien les exigences culturelles, techniques et légales du marché du maître d’ouvrage que les spécificités de l’aire culturelle ciblée : traditions, coutumes, particularités linguistiques, tabous, mots à la mode « cordes sensibles » des consommateurs. Ainsi, localiser signifie « traduire sur l’ordinateur, pour l’ordinateur » (cf. Delisle, 2021, p. 213). Avec la localisation, tout produit destiné à la commercialisation internationale (dont les plateformes numériques, logiciels, appareils électroniques...) conserve les mêmes valeurs d’usage dans les zones où il a été produit (zone de production), cependant, il apparaîtra sous une autre forme et la réussite ou l’échec de sa performance dans un autre foyer socio-culturel dépendra de la réussite ou de l’échec du modèle socio-culturel dans lequel on aura décidé de l’introduire. La transcréation n’est pas bien différente de la localisation. En réalité, les deux concepts sont très similaires. Pour n’importe quel exercice de traduction, transcréer signifie s’éloigner de l’original et partir d’une interprétation indicielle du texte source et ne se permettre aucune réversibilité ni même approximative (cf. Hammer, 2019, p. 236). Bien que le concept soit apparu au début en littérature et en poésie en tant que procédé de reformulation esthétisante qui consiste – particulièrement en poésie – à partir d’une image et/ou d’un ressenti pour recréer l’effet/émotion dans un autre vers afin de conserver l’essence volatile de l’original (cf. Lapeyre, Poulin, & Roger, 2011, p. 149) et qui préconise une traduction créative en privilégiant l’inventivité et la transposition délibérée du signifié textuel (Romengo, 2010, p. 175). Ce même concept a été associé à d’autres termes comme « versionisation » rattachés au domaine audiovisuel (cf. Rannaud, 2021, p. 112) et à toute une série de pratiques de transfert multilingue comme la rédaction technique, la médiation langagière qui prouvent que la traduction est actuellement une opération fluide, négociable suscitant représentations et attentes diversifiées (cf. Scarpa, 2010, p. 168 et Gambier, 2006, p. 30). Paratraduction, Transcréation et problématique de mise en tourisme en Alg érie (Tra vaux pratiques). La paratraduction unit les deux concepts tant au niveau de l’adaptation que celui de l’approche créative, cependant, une subtile différence dans le mode opératoire, dont l’importance n’est pas moindre, doit être mise en exergue. En effet, la localisation demeure un concept d’ensemble portant sur le traitement linguistique des « solutions informatiques » tels que les logiciels et les sites internet dont la traduction n’en est qu’un élément parmi une série d’adaptations culturelles. De plus, si en traduction ordinaire, si on travaille sur un texte déjà fourni, en localisation, la conception d’un produit de vente se fait simultanément avec son traitement linguistique. La transcréation, elle, est une activité purement créatrice qui peut intéresser les « sloganistes » ou les rédacteurs-publicitaires dans l’industrie du divertissement où il est toujours question de transposer efficacement un message dans une dimension créative tout en gardant l’esprit du concept de départ, par conséquent la transcréation demeure pleinement une créativité verbale. La paratraduction, quant à elle, inclut dans son exercice des parties d’un document à traduire lesquelles n’ont pas été prises en considération, comme le paratexte, l’image, les symboles et les sons, pourtant ceux-ci sont des indications de l’extra-texte qui ont toujours leur influence sur les statuts des textes traduits (cf. Yuste Frías, 2010, p. 294). 2.2.4.1 Critères d’évaluation Avant de procéder à l’analyse des deux plateformes, nous avons pris en considération trois critères d’analyse à savoir : « le plan ergonomique », « le plan de contenu », et « le plan multimodal », les portées techniques de chaque plan d’analyse sont les suivantes : 2.2.4.1.1 Le plan ergonomique Le plan ergonomique identifie les performances de la plateforme par rapport aux « lois de proximité et de similarité » ; la loi de proximité signifie que notre cerveau cherche à regrouper les éléments qui sont proches dans l’espace de lecture à l’écran. La proximité des éléments visuels avec les éléments verbaux et sonores exprime donc une cohésion du plan conceptuel général de la plateforme. En cas d’éloignement de ces éléments les uns des autres ou s’ils se présentent à nous de manière anarchique, cela sera interprété comme une « désorganisation cognitive » ce qui signifie que le lecteur a le sentiment d’être perdu dans les hypertextes qu’il parcourt soit à cause de la mauvaise régulation de l’information ou du manque de cohésion de la présentation soit à cause de l’hypercondensation du contenu sous certaines rubriques (Baccino, Salmerón et Cañas, 2008, p. 15). Toutefois, l’éloignement peut parfois s’avérer utile s’il prend la forme d’un petit espacement permettant de délimiter les blocs d’information. (Cf. Boucher, 2020, p. 47). Une qualité ergonomique d’une plateforme permet de distinguer les critères « d’efficacité, d’efficience et de satisfaction ». À travers ces critères on peut vérifier par exemple si les maquettes graphiques et fonctionnelles sont bien agencées. Pour l’interface qui illustre les deux, on peut examiner si celle-ci présente une bonne hiérarchie visuelle avec une bonne organisation des formes et des couleurs, des zones de respiration (les espacements) (cf. Boucher, 2020, pp. 17,18 et 28). Le plan ergonomique s’intéresse également au « niveau interactif » au moyen duquel on vérifie la manière dont les différents éléments du site invitent le visiteur à interagir avec le contenu (ibid., p.63). Les éléments d’interactivité concernent le plus souvent le web social : facebook, twitter, Instagram, YouTube qui traduisent une mise en visibilité des offres touristiques ; les dispositifs du web social connectés les uns aux autres favorisent la circulation des internautes au sein d’une architecture cohérente, et transforment même ces derniers en « ambassadeurs » de la promotion touristique, en associant leurs productions numériques à la valorisation du pays (cf. Jammet & Linder, 2019, p. 8). Le critère « d’utilisabilité et d’utilité » qui rentre également dans les options ergonomiques concerne le confort d’utilisation du site qui reflète sa valeur ajoutée. L’utilisabilité concerne la capacité d’un site web à répondre aux besoins d’informations de l’utilisateur et l’utilité est relative à la capacité de la plateforme d’être utilisée à bon escient pour l’information recherchée (Tricot et Bastien, 2008, p.215) ; l’utilisabilité, elle seule, se reflète également à travers des traits de l’architecture du site faisant référence à la bonne organisation de son contenu lesquels sont : organisation claire des rubriques ; optimisation de la recherche de l’information à travers un moteur de recherche interne ; page d’accueil intuitive ou contre-intuitive ; rubricage trop dense pour très peu de contenu ; chemin de navigation trop long ; design vieillissant comportant des choix techniques dépassés (cf. Rouquette, 2017, pp. 87-88). 2.2.4.1.2 Le plan de contenu Le plan du contenu s’intéresse à la méthode de présentation du site permettant son assimilation de manière fluide. Pour cela, un « écosystème de communication territoriale et culturelle » permet de vérifier si le site remplit sa fonction de faciliter l’accès et l’appropriation intellectuelle d’un patrimoine et s’il diffuse une image attractive d’une région touristique (Stockinger, 2017, p. 135) Ensuite, l’écosystème de communication territoriale et culturelle donne lieu à des analyses thématiques, narratives, rhétoriques et discursives qui sont des méthodes de recherche ciblées dans un contenu numérique qui permettent respectivement de dévoiler sa présentation générale, la posture de l’énonciation et du cadre énonciatif, les actes et les figures de rhétorique (ibid., p.143). Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). Le plan de contenu qui doit contenir un « objet de référence » concerne les stratégies mises en place par les concepteurs de la plateforme visant à anticiper les comportements des visiteurs potentiels en traçant des objectifs sur le type d’internautes visés, leurs besoins (supposés), leurs attentes programmées, la stratégie d’argumentation préconisée, la rubrique du site choisie pour être mise en avant plus que d’autres. Si les stratégies de l’objet de référence s’avèrent concluantes, cela donnera un contenu qui sera présenté à l’internaute dans le style de rédaction qui lui sied, avec plusieurs choix interactifs (transaction, visionnage, accès à un contenu de fidélité etc.) ainsi qu’une possibilité de choix de navigation (cf. Rouquette, 2017, pp.82-88). Une « rhétorique promotionnelle » s’appuie, elle, sur la logique de la distinction du produit vanté. Elle concerne spécifiquement « la portée du paysage textuel » (ensemble d’interactions linguistiques avec des notions culturelles et la « performance scénique » c’est-à-dire que la performance du texte est évaluée par l’effet qu’il produit sur le lecteur, par l’accomplissement de son action autant qu’il est le « sujet même » de sa présentation. 2.2.4.1.3 Le plan multimodal Le plan multimodal est un plan imagé, sonorisé et textualisé, il reflète une multiplicité des interactions verbo-iconiques qui mettent en valeur l’esthétique de la plateforme numérique et renforcent sa lisibilité étant donné que, fonctionnellement, toute page numérique utilitaire s’assimile facilement à travers l’ensemble de mises en forme les unes à côté des autres qui dessinent un tout (Boucher, 2020, p. 46). Pour Stockinger, l’expertise sémiotique d’un site web repose sur la cohérence de son contenu qui se reflète par des strates (modules d’affichage) ordonnées lesquelles renferment les thématiques développées du site. Les différentes strates qui constituent une présentation portant sur un objet de référence (région touristique) relié à un thème, un parcours syntagmatique, une expression verbale, une mise en scène, et à une réalisation médiatique (vidéo) (Stockinger, 2017, p.150), tout cet enchaînement est reconnaissable par celui qui consulte la plateforme sous forme d’une hyperréalité. Il existe des situations de relations intersémiotiques où l’on peut avoir un topos spécifique qui souligne un univers de sens suffisamment attirant aux yeux du public concerné pour qu’il soit poussé à agir. La relation intersémiotique peut être exprimée différemment, cela dépend des variations de sens selon le type de l’image et de la rhétorique textuelle. Ces variations ont un impact sur le destinataire, sur ses attentes et sa volonté d’accepter ce contrat offert par la plateforme. (cf. Stockinger, 2017, p.149-150), c’est-à-dire la façon dont ses modalités d’énonciation donnent forme à un dispositif qui assigne une « place » et un « parcours » à la figure du destinataire qu’il élabore (cf. Jammet & Linder, 2019, p. 6). duction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). Le plan multimodal peut par exemple privilégier la mise en valeur de l’identité, car elle constitue un élément incontournable dans l’univers du tourisme. Lorsqu’on consulte une plateforme touristique d’un pays, d’une métropole ou d’une région, il est important de ressentir à travers les ressources textuelles et iconiques déployées dans cette plateforme qu’ils représentent l’identité du pays visité (Amirou, Pauget, & Lenglet et al. 2011, p.89). 2.2.4.1.4 www.algeriatours.dz La plateforme touristique « Algeriatours.dz » est la plateforme officielle de la promotion touristique en Algérie. Lancée en 2022 par le ministère du tourisme et de l’artisanat en remplacement d’une autre plateforme « visitalgeria.dz » créée en 2018 devenue obsolète à cause de l’échec d’une refonte du site et de l’absence d’un cycle d’amélioration continue à travers l’apport des microchangements. L’objectif du ministère du tourisme algérien à travers « Algeriatours.dz » est de mettre en place une vitrine pour le tourisme algérien qui réponde aux exigences du marketing virtuel à l’échelle mondiale et en ce sens, la voie numérique a été donc consacrée pour donner un avant-goût sur les vestiges du paysage algérien et de sa diversité. Nous avons choisi de nous concentrer dans l’analyse du site, sur les principales ressources sémiotiques distribuées dans son interface et de voir si celles-ci sont suffisamment bien combinées pour construire un imaginaire touristique au moyen d’un contenu hyperréel et si, dans l’hypothèse d’une paratraduction de cette plateforme en plusieurs langues, le traducteur peut s’appuyer sur les ressources non verbales pour appliquer un tel concept. Figure 71Plan d'accroche de la page d’accueil de la plateforme algeriatours.dz La plateforme est construite sur huit modules d’affichage (strates), sept rubriques de services touristiques composent le premier module avec un plan d’accroche (cf. figure 71) (page d’accueil, circuits et sites touristiques, restauration, hôtellerie. Le plan d’accroche invite le visiteur à découvrir les principaux circuits touristiques du pays en proposant un menu dans lequel des circuits à thème (culturels, historiques, sahariens et balnéaires) sont à sélectionner. de en Algérie (Travaux pratiques). Derrière le plan d’accroche, un système de défilement d’images en boucle semble renforcer le slogan traduit en français et en anglais : Ara.‫اكتشفوا أجمل المواقع الأثر ية وأروع المعالم السياحية وأجمل المناظر الخلابة في رحلات على مدار السنة‬ Fra Découvrez les plus beaux endroits, les meilleures attractions et les merveilleux voyages toute l’année. Discover the most beautiful places, the best attractions and wonderful trips all year round. Le slogan avec ses traductions correctes forme avec les images en mouvement et en arrièreplan, une homogénéité sémantique. On peut dire que l’on obtient à ce niveau une seule interaction verbo-iconique. Cependant, cette interaction pouvait être esthétiquement plus accentuée si les concepteurs avaient employé des images plus évocatrices, ou fantaisistes par rapport au but recherché, un tel exemple nous laisse perplexe quant à son adéquation au processus de paratraduction. En dépit de cette relation intersémiotique, le texte et les images en mouvement sont loin d’être complémentaires pour former un langage unidimensionnel au sens d’Albert Plécy (cf. supra, p.347). Les images, de par leur nature, ne semblent s’associer, dans le slogan, à aucune allusion verbale précise qui en traduirait de manière légitime leur pouvoir figuratif. Autrement dit, le slogan n’est pas assez métaphorisé ou puissant pour faire « parler » les images. Tout ce qu’on retiendra des occurrences » ‫ « أروع المعالم‬et » ‫« أجمل المناظر الخلابة‬ c’est qu’elles relèvent d’opérations déductives de lecture de l’image par décodage verbal, on peut donc se passer carrément du texte. En ce qui concerne la stratégie de communication numérique, on constate que la plateforme se présente plutôt comme un inventaire ou un annuaire à travers l’indication de 380 circuits répartis sur 58 villes pour un total de 1147 lieux touristiques. Il s’agit là d’une erreur de stratégie, puisque l’on s’attend à trouver dans une plateforme un contenu relativement bref avec une argumentation qui allie mystère, curiosité, beauté et originalité. De plus, tout visiteur ne sera pas tenté de parcourir tous les circuits proposés à la manière de consultation d’un a Sur le plan du contenu, les textes de présentation des villes et des circuits sont construits selon la progression thématique suivante : situation géographique, histoire et atouts de la région visitée. Cependant au niveau de l’énonciation, les stratégies d’argumentation qui devraient amorcer un rapprochement avec les visiteurs virtuels à travers les figures rhétoriques et les fonctions d’illocution et de perlocution sont peu nombreuses. Au niveau de l’affichage, le texte touristique est largement distancé de l’image qui l’accompagne et qui devrait lui servir de E Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). paratexte – plus précisément son péritexte iconique – cet éloignement annihile leur interaction verbo-iconique et les deux exemples suivants permettent d’étayer notre constat : Exemple 01 : Figure 72 Texte introduisant la ville de Bejaïa (Algérie), source : algeriatours.dz Ara ‫ من‬،‫ ولها حدود جد استراتيجية حيث يحدها من الشمال البحر الأبيض المتوسط‬2‫كلم‬3.268 ‫تبلغ مساحة ولاية بجاية‬ ‫ ومن الجنوب الغربي ولاية برج‬،‫ ومن الغرب ولاية البويرة‬،‫الشرق ولاية جيجل ومن الجنوب الشرقي ولاية سطيف‬.‫ وهي شديدة البرودة في فصل الشتاء‬،‫ ومن الشمال الغربي ولاية تيزي وزو‬،‫بوعر يرج‬ ‫ وتتوفر على الحظيرة الوطنية ڤ ورايا والمصنفة‬،‫ به العديد من غابات الصنوبر وأشجار الزيتون‬،‫تتمتع الولاية بغطاء نباتي متنوع‬ ،‫ وهي تصنف ضمن الولايات السياحية الأكثر مقصدا‬،‫ بالإضافة إلى شر يط ساحلي خلاب وميناء صيد وآخر تجاري‬،‫عالميا‬ ‫ إذ تعتبر من أجمل ولايات الوطن واكثرها جذبا للسواح و يعتبر طبق ال كسكس من الأكلة‬،‫لثرائها الحضاري والأثري‬ َّ َّ.‫الأكثر شعبية بالمنطقة‬ Fra Ang Bejaia a une superficie Couvrant une superficie de 3.268km2. Bejaia occupe une position stratégique. bordée Bordée au nord par la mer Méditerranée, à l’est par la wilaya de Jijel Jijel, au sud-est, la wilaya de Sétif par Sétif, à l'ouest par e Brouira Bouira, au sud-ouest par Bordj Bou Arreridj, et au nord-ouest par Tizi Ouzou. Région très froide en hiver. Réputée pour son hiver très froid La wilaya Bejaia possède un couvert végé tal très dense, de nombreuses forêts de pins et d'oliviers. Elle dispose d'une réserve nationale classée mondialement ; Gouraya, en plus d'une en tant que bande côtière pittoresque, d'un surplombant port de pêche et d'un port de commerce un port multifonctionnel. Bejaia est classée parmi les wilayas villes touristiques les plus visitées pour sa richesse culturelle et ses zones archéologiques. Elle est considérée comme l'une des plus belles régions du pays et reès attractif très attractive pour les touristes. Le couscous est le plat le plus populaire de la région. On y déguste le couscous comme plat le plus populaire de la région. Bejaia covers an area of 3,268km2. It occupies holds a strategic position, bordered to the north by the Mediterranean Sea sea, to the east by the province of Jijel, to the south-east by the 389 QUATRIEME PARTIE Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). province of Sétif, to the west by Brouira Bouira, to the south-west by Bordj Bou Arreridj, and to the north-west by Tizi Ouzou. The region is very cold in winter. This province Bejaia has a very dense landscape, with numerous plenty of pine and olive forests. It has a world-class national reserve, Gouraya, as well as a as it is a scenic coastal strip, a fishing port and a commercial port overlooking a multipurpose port. The province is ranked among the most visited tourist wilayas cities for its cultural wealth heritage as well as its archaeological areas. It is regarded as one of the most beautiful regions of the country and very appealing to tourists. Its most popular cuisine is Couscous. Hormis les erreurs soulignées ci-dessous que nous avons corrigées, les versions anglaise et française du texte présentant la ville de Bejaïa sont plutôt correctes. Cependant, on remarque une absence totale de marqueurs du discours argumentatif censés illustrer l’attractivité de la ville, le texte est plutôt informatif et descriptif (distribution des éléments du réel dans l’espace hors du temps et transmission neutre d’information) et la photo placée au-dessus qui montre une ancienne bâtisse, celle du fort Moussa, n’apporte aucune valeur esthétique en tant que source d’argumentation de relais par rapport au contenu développé. (cf. figure 72). Exemple 02 : Figure 73 Texte introduisant la ville de Constantine, source : algeriatours.dz Ara.‫ معالم تناطح الزمان والمكان‬،‫ أصالة العمران‬،‫قسنطينة عبق الثقافة‬ ‫ فهي قطب سياحي بامتياز لما‬،‫ مدينة تتميز بسحر خاص مما جعلها من أهم وجهات السياحة في الجزائر‬،‫قسنطينة السياحية‬ ‫ هي تلك المدينة المعلقة على ضفتي‬،‫تحتو يه من مقومات سياحية وعرض سياحي ثري ومتنوع ممتد عبر الأزمنة والعصور‬ ‫صخرين عتيقين ربطا بجسور زاوجت بين العراقة وا لأصالة والمقاومة من جهة وبين مفخرة النصر والتمدن والمعاصرة من‬.‫جهة أخرى‬ 390 QUATRIEME PARTIE Paratraduction, Transcréation et problématique de mise en tourisme en Algérie (Travaux pratiques). ‫ سنة فتركت فيها آثارا زادت من عراقتها وامتدادها في التاريخ وعبر‬2.500 ‫هي مدينة تداولت عليها الحضارات لما يربو عن‬ ‫ مجلد بعنوان قسنطينة حكاية الزمان‬،‫ فهي ذلك المجلد الذي تجد فيه عند التنقل عبر صفحاته حكاية لكل موقع‬،‫العصور‬.‫والمكان‬.‫هي مدينة تيديس الأثر ية أو مدينة الأقادسة ل كثرة الديانات والمعتقدات بها‬.‫هي المدينة العتيقة التي تعد إرثا معنو يا وجماليا هاما وذلك من خلال بيوتها المسقوفة ومنازلها ذات الطراز المعماري المميز‬ ‫هي مدينة الأسواق والسو يقات والأزقة التي لا تزال إلى الآن تحمل اسم الأنشطة التجار ية التي كانت تزاول فيها قبل وإبان‬ ‫ وهي‬،‫ إلخ‬.... ‫ دار الدباغ‬،‫ فندق الز ياتة‬،‫ مقعد الحوت‬،‫ الجزارين‬،‫ رحبة الجمال‬،‫الحقبة الاستعمار ية كرحبة الصوف‬.‫ال كهوف الأثر ية التي تعود إلى عصور ما قبل التاريخ‬ ‫هي ضريح ماسينيسا ذلك البرج المربع المشيد على شكل مدرجات وصفوف من الحجارة المنحوتة بصورة مستوحاة من‬.‫الأسلوب الإغريقي البونيقي وهي الأقواس الرومانية الشاهدة على الحضارة الرومانية‬ ،‫ فهي قسنطينة النحاس‬،‫ تشكل في مجملها هو ية سكانها‬،‫هي زهرة الصناعات التقليدية التي تجد في كل بتلة من بتلاتها حرفة‬ ‫ قسنطينة الورد والزهر‬،‫ قسنطينة الأطباق التقليدية الغنية بالأذواق‬،‫ قسنطينة المالوف‬،‫قسنطينة الألبسة التقليدية والنسيج‬...‫المقطر‬ ‫ إنها قسنطينة‬...‫نعم يا سادة‬ Fra Ang Constantine, le sens de la culture senteur de culture diffus e, l'authenticité de l'urbanisation symbole d’un art architectural authentique, les contours du temps et de l'espace une ville magnifiée au fil des ères. Constantine touristique, l’envoûtante, est une ville caractérisée par son propre charme au charme particulier, qu’il l’a la rend une des destinations touristiques les plus importantes d'Algérie. C'est un pôle touristique par excellence du fait de ses potentielles touristiques son potentiel touristique et d'une offre touristique de ses dotations factorielles aussi riches et que diversifiées qui traverse se prolongent au fil dles époques et les des âges. C'est elle la ville suspendue sur les rives de deux anciens rochers reliés par des ponts qui allient concilient tradition, originalité et résistance d'une part, et fierté de la victoire, urbanisation urbanité et modernité d'autre part. C'est une ville qui a vécu plusieurs civilisations depuis plus Constantine existe depuis de 2500 ans et a connu plusieurs civilisations qui ont laissées des tracesleurs empreintes témoigner de sa noblesse et de son ancestralité et son extension dans l'histoire et à travers les âges. C'est ce Elle est ce livre dans lequel en naviguant à travers sessi vous vous aventurez, vous trouverez une histoire pour chaque site lieu un livre, ce livre étant intitulé : Constantine l'histoire du temps et du lieu. C'est la ville archéologiqueElle est « Tidiss », la ville archéologique ou la ville des saints en raison du grand nombre de religions et de croyances qui s'y sont croisées. La C’est la vieille ville de Cirta, considérée comme un important patrimoine moral immatériel et esthétique, à travers ses maisons au style architectural distinctif. C'est la ville des marchés, des « souikats » et des ruelles qui portent encore le nom des activités commerciales qui s'y pratiquaient avant et pendant l'époque coloniale, comme « rahbet sof » (marché de la laine) « rahbet lejmel » (marché des chameaux) « El -jazarine (des bouchers) mak’âdd elhout (place du poisson), Foundok ezziata (des vendeurs d’huiles), Dar Al-Dabbagh (maison de la maroquinerie) etc. C'est la ville des grottes archéologiques qui remontent à la préhistoire. C’est le tombeau de la ville où repose Masinissa, cette tour carrée construite en forme de terrasses et de rangées de pierres sculptées dans une image inspirée du style grécopunique, et les arcs romains témoignent de la civilisation romaine... Elle est ce joyau de l’artisanat aux éclats multiples et colorés qui font jaillir ses métiers divers telle une floppée de couleurs flamboyantes qui reflètent dans une mosaïque l’identité de sa population, c’est Constantine, ville du cuivre, ville d’habits traditionnels, ville du métier à tisser, ville du malouf, de gourmandise traditionnelle copieuse, ville des roses et de leurs nectars. Oui, messieurs... c'est Constantine! Constantine, cultural flair is of uplifting cultural scent, authenticity of urbanization a symbol of authenticity and urban remarkable style, contours of time and space crafted by time and space. Constantine is a charming city, characterized by its own unique appeal, which has made it one of the most significant relevant tourist destinations in Algeria. It is a tourist pole par excellence therefore an ideal place for tourism because of thanks to its tourist rich potential, a its and diversified welfare conditions and welcoming offers tourist offer which crosses the times and the agesinherited since ages. It is thate city suspended on the banks of two ancient rocks linked by bridges which combine tradition, originality and resistance on the one hand, and pride of victory, urbanisation urbanity and modernity on the other. It is a city that has experienced several civilizations over the last 2500 years, leaving traces that have increased its nobility and extended it extension in history and through the ages. It is this that book in which, as you if browse through its pages you go wandering through its pages, you will find a story for each site place, a book that may be entitled Constantine: the a history of time and space. It is the archaeological city "Tidiss" or the city of saints because of the great number of religions and beliefs that have crossed there. The It is the old town of Cirta, considered as an important moral and aesthetic heritage, through its houses with a distinctive architectural style. It is the city of markets, called "souikats" and alleys that still bear the names of the commercial activities that took place there before and during the colonial period, such as "rahbet sof" (wool market), "rahbet lejmel" (camel market), "El -jazarine" (of the woodsheds), "mak'âdd elhout" (fish square), "Foundok ezziata" (oil sellers), "Dar Al-Dabbagh" (house of leatherwork), etc. It is the city of archaeological caves dating back to prehistoric times.	33,840
52/hal.parisnanterre.fr--hal-01543833-document.txt_2	French-Science-Pile	Open Science	Various open science	null	None	None	French	Spoken	1,745	2,744	417 Waldeck a copié à Mexico des statues semblables, associées au culte de Xipe Totec. Ce « soldat de la mort », en tant que tel, est inventé, et son frère jumeau s'était déjà glissé, sans légende ni explication, dans une planche de son ouvrage consacré à Palenque. Source : Dé tails du table au de Waldeck Le sacrifice gladiatorial. Photographie : Museo Soumaya. rement à la mort, au sacrifice humain, aux ossements et aux crânes, de fait très présents comme motifs ornementaux en Mésoamérique. Cet intérêt est aussi manifesté deux fois à l'Académie de San Carlos au cours de la seconde moitié du siècle. Petronilo Monroy, probablement né dans la décennie de 1830 et décédé en 1882, laisse à la postérité un tableau qu'il n'a jamais présenté aux expositions académiques : El sacrificio de una princesa acolhua ; la scène, peinte dans des tonalités extrêmement sombres, se réfère à un épisode de l'histoire ancienne des Mexicas, à peu près contemporain de la fondation de Tenochtitlan. Le souverain des Acolhuas, pour lesquels les Mexicas travaillaient, accorde sa fille en mariage au peuple aztèque ; lorsqu'il se rend aux festivités, sa fille a déjà été déifiée, c'est à dire sacrifiée et écorchée, et devant sa dépouille danse un prêtre revêtu de sa peau. Monroy fait l'impasse sur le rituel et représente simplement le père reculant d'effroi en voyant sa fille morte, allongée sur un autel. La tonalité extrêmement sombre et froide du tableau et l'attitude de recul du personnage laissent supposer un jugement moral négatif. Adrián Unzueta, auteur du tableau déjà commenté Moctezuma reçoit sa nomination, expose en 1898 El Tzompantli, une oeuvre diamétralement opposée à la précédente ; il est alors professeur à l'Académie, et non plus étudiant. La scène représente des prêtres aztèques en train de décapiter les corps de sacrifiés espagnols pour compléter un « tzompantli » ou tour de crânes. Aucune identification n'est possible, ni avec cadavres des Espagnols, ni avec les prêtres ; il ne s'agit pas non plus de la distance du savant face à l'objet de son étude, mais bel et bien d'une sorte de voyeurisme. Le journal d'opposition El Hijo de El Ahuizote somme l'artiste de représenter plutôt les tortures que les Conquistadors ont fait souffrir aux Indiens, et non l'inverse, et El Mundo Ilustrado, journal subventionné, caricature entre autres ce tableau. Unzueta, qui adopte un point de vue aussi extérieur et fasciné que Waldeck, est clairement en décalage avec les attentes du public aisé mexicain de la fin du XIXe siècle, qui attend des prises de Un tableau mexicain qui heurte la critique, en représentant position et une condamnation morale la cruauté systématique des Aztèques envers les Espagnols. Adrián Unzueta : El Tzompantli (Sacrificio de Españoles por de la violence. Mexicas). S'il ne permet pas aux Mexicains de s'enorgueillir d'une culture originaire de leur territoire national, Waldeck ne justifie pas non plus la situation de subordination dans laquelle sont tenus les Indiens. En 1869, avant de commenter son tableau d'histoire, l'artiste prend la défense des Indiens et attaque indirectement les gouvernements mexicains successifs. Les pages 4 à 8 sont consacrées à l'« anthropologie des races d'Amérique du centre ». Après un éloge de la « nation chole », originellement blanche, dont la déviation de l'angle facial (nez aquilin, front fuyant) serait selon lui naturelle, et non provoquée par des tablettes, Waldeck commente les variations de taille, la musculature prononcée, l'adresse, et finalement l'intelligence des Indiens, égratignant au passage « les Espagnols, qui prétendirent que l'Indien était une brute, incapable de devenir « même raisonnable », réservant ce dernier titre pour eux-mêmes! [] ». Il cite en contre-exemple « [] un élève indien [de Mexico] qui, au bout de dix-huit mois, dessinait très-correctement (sic) et avait appris en même temps l'anglais et le français », puis « un Indien pursang, à moustaches, nommé Patiño, lequel était peintre, sculpteur égalant Bozio ou Certellier, excellent mathématicien et polyglotte.[] Et pourtant ce grand artiste, je dirai presque ce savant, n'a été employé par aucun des nombreux gouUn peintre français réinvente le Mexique ancien 419 vernements qui se sont succédé à Mexico! 60 ». Il semble ignorer la quasi-absence de mécénat d'État à cette époque troublée et la fonction de directeur de l'Académie des Beaux Arts de San Carlos occupée par Patiño pendant quelques s. Cette partie se conclut ainsi : « Ma dernière preuve, enfin, Juárez est Indien, je l'ai connu jeune à la Mineria. Son instruction seule l'a conduit à la présidence 61 ». Cette référence au Président de la République parachève la démonstration. En revanche, en 1838, il était très ambigu dans son attitude face aux Indiens duYucatán et il n'écartait pas à leur égard la possibilité d'une infériorité naturelle. Tandis qu'il fait déjà l'éloge de leurs talents artistiques 62, il observe aussi que les Yucatèques présentent des caractéristiques qui les rapprochent du singe : « Les femmes ne se baissent jamais pour ramasser quoi que ce soit à terre ; avec les doigts du pied elles enlèvent un fruit, un morceau de bois, et jusqu'à une pièce de monnaie, et quoique ayant un panier plein sur la tête. Il peut à la fois faire l'éloge de la chaise à porteurs comme moyen de locomotion (il la représente dans un tableau exposé au salon de 1870) 64 et se scandaliser du fait que les Indiens soient vendus comme esclaves, sans pourtant se cacher d'en avoir lui-même acheté pour ses besoins personnels 65. Il écrit aussi : « Leurs « Et pourtant ce grand artiste, je dirai presque ce savant, n'a été employé par aucun des nombreux gouvernements qui se sont succédé à Mexico! » J.-Fr. M. Waldeck, Le sacrifice gladiatorial, op. cit., p. 7-8. 61 Ibidem. 62 « Il est à remarquer que les Meridaños, qui n'ont pas la moindre idée des arts, et qui sont surtout étrangers au dessin et aux principes de la sculpture, exécutent d'une façon très originale et fort régulière la charge de l'objet dont ils veulent rendre l'image. L'anatomie à part, les formes et les proportions de leurs oeuvres sont parfaitement justes ; et, chose encore plus surprenante, un seul outil, c'est-à-dire, un tronçon de sabre, fixé dans un manche de bois, suffit à ces rudes artistes pour animer leurs blocs de bois ou de pierre. En cela, les ouvriers de l'(sic) Yucatan ressemblent à ceux de l'Égypte, qui, avec des moyens à peu près semblables, font des travaux que ne renieraient pas des gens plus habitués aux procédés de l'esthétique. L'adresse des ouvriers yucatèques se révèle aussi dans la confection des instruments de musique []. Il ne manque à ce peuple que la tranquillité et un gouvernement qui encourage et développe ces dispositions naturelles []. À l'époque de mon arrivée dans la capitale de l'Anahua , les arts manuels étaient dans l'enfance ; des ouvriers étrangers vinrent s'y établir et exercer leurs professions. Quand j'ai quitté Mexico, les artisans indigènes étaient aussi habiles que leurs maîtres ; ce fait n'est-il pas significatif? J-Fr. M. Waldeck, Voyage pittoresque et archéologique, op. cit., p. 34. 63 J-Fr. M. Waldeck, Voyage pittoresque et archéologique dans la province d'Yucatan, op. cit., p. 65-66. 64 « La manière de voyager dans l'intérieur de l 'Yucatan se rapproche de celle qui est en usage dans les Indes orientales. On est porté par des hommes dans une litière couverte qu'on appelle cochè (prononcez cotchai ). Quand on est habitué à cette espèce de palanquin , on le préf ère au cheval ». Ibidem, p. 67. Conclusions Dans l'état actuel des connaissances sur le sujet, il semble que les points communs entre les productions mexicaines et le tableau de Waldeck soient plutôt des coïncidences que le résultat de contacts. Les contextes de production étant radicalement différents, on ne peut sans doute pas attribuer les mêmes intentions aux artistes de l'un et l'autre pays. Cela n'empêche pas la présence d'un contexte culturel commun ; la mise en parallèle du corpus mexicain avec le tableau français, possible dans la mesure où la culture de l'élite mexicaine est sensiblement identique à celle des élites européennes, permet plusieurs constats. D'une part, les discordances, qui n'apparaissent pas à première vue, sont multiples. Waldeck est ouvertement républicain et méfiant vis-à-vis des monarques, ce qui peut prêter à sourire sous le règne de Napoléon III ; mais il ne considère pas son objet d'étude comme un idéal et pose un regard ambigu sur les Aztèques, à la fois universaliste, – puisque Tlahuicole incarne des valeurs de patriotisme et de courage –, et voyeur, insistant sur le caractère sanglant de cette civilisation, sujet qui sera aussi traité au Mexique, avec bien plus de violence. La principale leçon réside sans doute dans l'ampleur du parallélisme entre le tableau de Waldeck et le corpus académique du dernier tiers du siècle : cette concordance poussée permet de voir d'une part la diversité des points de vue et intentions des artistes académiques eux-mêmes, de Parra, qui utilise les souffrances des Indiens pour galvaniser les Mexicains contre les impérialismes, à Unzueta, capable à la fois de faire l'éloge de l'Empereur et donc du dictateur, et de rappeler la distance qui sépare les sanguinaires Aztèques des Mexicains moderne . Mais elle montre d'autre part à quel point le regard des artistes académiques est distancié, et dans quelle mesure la culture et l'esthétique européenne sont assimilées et assu66 J-Fr. M. Waldeck, Voyage pittoresque et archéologique dans la province d'Yucatan, op. cit., p. 43. 421 mées comme propres. Entre un artiste français utilisant principalement sa culture livresque et un Mexicain qui représente le patrimoine de son propre pays et les ancêtres de ses concitoyens, les différences sont somme toute minimes, en partie parce que les artistes académiques ne reçoivent pas au XIXe siècle de cours d'archéologie « nationale ». Pour les Mexicains comme pour le Français, l'archéologie est un faire-valoir et les Indiens sont un objet d'étude. Ils chassent en quelque sorte sur le même territoire, ce qui a d'abord valu à Waldeck l'appui d'Alamán au moment de concrétiser son projet d'exploration de Palenque, mais a provoqué par la suite les tensions avec le gouvernement de Santa Anna en 1836. Si Waldeck publie ses notes de voyage en 1838, c'est avant tout, il ne s'en cache pas, pour éviter que les résultats de ses recherches ne soient publiés par le gouvernement mexicain, lui enlevant la gloire qui lui revient. Il y a donc, en un sens plus, de dissonances entre les voix mexicaines qu'entre les deux continents..	47,401
W2976102486.txt_1	Open-Science-Pile	Open Science	Various open science	2,019	Spin-driven electrical power generation at room temperature	None	Unknown	Unknown	7,136	13,427	ARTICLE https://doi.org/10.1038/s42005-019-0207-8 OPEN 1234567890():,; Spin-driven electrical power generation at room temperature K. Katcko1, E. Urbain1, B. Taudul1, F. Schleicher1,2, J. Arabski1, E. Beaurepaire1,5, B. Vileno D. Lacour 2, S. Boukari1, M. Hehn2, M. Alouani1, J. Fransson4 & M. Bowen 1 3, D. Spor1, W. Weber1, On-going research is exploring novel energy concepts ranging from classical to quantum thermodynamics. Ferromagnets carry substantial built-in energy due to ordered electron spins. Here, we propose to generate electrical power at room temperature by utilizing this magnetic energy to harvest thermal ﬂuctuations on paramagnetic centers using spintronics. Our spin engine rectiﬁes current ﬂuctuations across the paramagnetic centers’ spin states by utilizing so-called ‘spinterfaces’ with high spin polarization. Analytical and ab-initio theories suggest that experimental data at room temperature from a single MgO magnetic tunnel junction (MTJ) be linked to this spin engine. Device downscaling, other spintronic solutions to select a transport spin channel, and dual oxide/organic materials tracks to introduce paramagnetic centers into the tunnel barrier, widen opportunities for routine device reproduction. At present MgO MTJ densities in next-generation memories, this spin engine could lead to ‘always-on’ areal power densities that are highly competitive relative to other energy harvesting strategies. 1 Institut de Physique et Chimie des Matériaux de Strasbourg, UMR 7504 CNRS, Université de Strasbourg, 23 Rue du Lœss, BP 43, 67034 Strasbourg, France. Jean Lamour UMR 7198 CNRS, Université de Lorraine, BP 70239, 54506 Vandœuvre les Nancy, France. 3 Institut de Chimie, UMR 7177 CNRS, Université de Strasbourg, 4 Rue Blaise Pascal, CS 90032, 67081 Strasbourg, France. 4 Department of Physics and Astronomy, Uppsala University, Box 516, 75120 Uppsala, Sweden. 5Deceased: E. Beaurepaire. Correspondence and requests for materials should be addressed to M.B. (email: bowen@unistra.fr) 2 Institut COMMUNICATIONS PHYSICS \| (2019)2:116 \| https://doi.org/10.1038/s42005-019-0207-8 \| www.nature.com/commsphys 1 COMMUNICATIONS PHYSICS \| https://doi.org/10.1038/s42005-019-0207-8 a EF 2 ΓR↓ ΓL↑ ΓR↑ PM center Spintronic selector L Spintronic selector R b E Δ EF ↑ ↓ FM electrode Spinterface PM center c Spinterface Tunnel barrier Spinterface FM electrode FM electrode E PM center EF Results Figure 1a illustrates our spin engine in the simpliﬁed case of a PM center, characterized by two effectively spin-split energy levels. To achieve a strongly spin-dependent transmission rate Γ, the PM center is placed between spintronic selectors—materials systems that ideally favor only one transport spin channel while blocking the other. Examples include half metals15–17, 2D materials with half-metallic properties18, a normal metal / ferromagnetic tunnel barrier19 bilayer, and the ferromagnetic metal/molecule interface13, also called a ‘spinterface’. The Fe/MgO system may also constitute a spintronic selector given either a sufﬁcient MgO thickness20 and/or the presence of oxygen vacancies21–24. A standard ferromagnetic metal (FM) would also work, albeit with reduced efﬁciency. Due to this combination of spintronic selectors and spin-split PM states, a spin ↑(↓) electron may only depart the PM toward the left(right) electrode at the energy of the PM center’s corresponding spin state. Hai et al.1 used a MnAs ferromagnetic metal with a conventional (~50%) spin polarization of conduction states, and applied a magnetic ﬁeld to spin-split their MnAs QDs and obtain power generation at 3K. It is unclear whether this experiment could have worked at higher temperatures. Miao et al.2 ﬁltered the electron spin upon transport across EuS ferromagnetic tunnel barriers at 1K, i.e. below its ordering temperature TC ~ 16.8 K. Here, spin splitting of the Al QD is induced by electronic coupling to one EuS barrier. Reports indicate that several spintronic selector tracks include materials science candidates (e.g. the Fe/MgO MTJ class20,22, the half-metallic Co2FeAl17 or the ferromagnetic tunnel barrier CoFe2O419) that can operate at/beyond room temperature (RT). ΓL↓ kBT A solar cell’s electronic potential landscape is crafted such that, when a photon is absorbed, the resulting electron and hole (the absence of an electron) ﬂow in opposite directions. Since they carry an electrical charge of opposite sign, this generates an electrical current. Two low-temperature experiments1,2 have suggested that, by astutely designing the magnetic potential landscape of a quantum dot (QD) device, electrons with a spin ↑ or ↓ quantum property can ﬂow in opposite directions. This can generate electrical power if the spin ↑ and ↓ current channels are imbalanced, i.e. if the overall current is spin-polarized. This apparent current imbalance, and the presence of QDs in both systems, are reminiscent of quantum thermodynamical experiments on single electron boxes, which have demonstrated how to harvest thermal ﬂuctuations3,4 and information5 to perform work at very low temperatures. These heat and information engines are driven by ﬂuctuation-induced quantum tunneling on/off of QDs, with a transmission asymmetry between left and right leads that can be energy-dependent due to the QD’s discrete energy levels6,7. A few reports have theoretically8 and experimentally (using nitrogen vacancies in diamond9) taken into account the electron spin. Inspired by the report of Miao et al.2, and by recent progress in quantum thermodynamics3–12, we propose that a spin-split paramagnetic (PM) quantum object can enable electrons with a spin ↑ or ↓ quantum property to ﬂow in opposite directions if the transmission rates on either side of the PM center are spindependent. Differing amplitudes in these transport spin channels generate a spontaneous current ﬂow. Measurements across a single MgO magnetic tunnel junction, backed by analytical and ab-initio theories, indicate that this spin engine can operate at room temperature. We discuss strategies to achieve routine device reproduction. Our work also conﬁrms the high transport spin polarization at room temperature of the ferromagnetic metal/ molecule interface13 inferred from spectroscopy measurements14. k BT ARTICLE ↑ Δ EF ↓ Fig. 1 A room-temperature spin engine. a Illustration of asymmetries in the spin-induced transmission rates Γ across a single paramagnetic (PM) center between spintronic selectors L and R. Spinterfaces13,14,25,26, halfmetals15,17, Fe/MgO MTJs20,22, and ferromagnetic tunnel barriers19 can fulﬁll the role of a spintronic selector. b Spin-conserved quantum tunneling between a spinterface and a PM center deforms the PM center’s Bloch sphere, thereby splitting27,28 its spin states by Δ, and shifts the spinterface’s Fermi level (EF) by Δϕ. c The spintronic landscape across a magnetic tunnel junction, comprising spinterfaces and a PM center, in its antiparallel (AP) magnetic state exhibits a spontaneous bias voltage ΔV ≤ Δ. A yellow band designates the thermally rectiﬁed, spin-polarized current involving striped segments of the ferromagnet (FM) and spinterface density of states. The spin engine generates work by harvesting thermal spin ﬂuctuations (kBT) on the PM center. See text for details. In all panels, red (blue) correspond to spin ↑(↓) states To obtain RT electrical generation, and in the process demonstrate it to be a RT spintronic selector, we utilize the spinterface13,14,25,26. This refers to a low energy bandwidth, low density of highly spin-polarized states that arise at room temperature from spin-polarized hybridization between the highly degenerate electronic states of a FM metal such as Co and the few, energetically discrete states of molecules, including carbon atoms26. The spinterface is weakly conducting, and its magnetic orientation naturally follows that of the FM metal. To date, only spin-polarized photoemission spectroscopy14,26 suggests that the spinterface may be a spintronic selector at RT. COMMUNICATIONS PHYSICS \| (2019)2:116 \| https://doi.org/10.1038/s42005-019-0207-8 \| www.nature.com/commsphys ARTICLE COMMUNICATIONS PHYSICS \| https://doi.org/10.1038/s42005-019-0207-8 a b 40 T = 295 K V = 10 mV TMR = –17% 200 Current (nA) Current (nA) 210 190 180 0 0 1000 2000 AP –2000 –1000 0 External magnetic field (Oe) T = 295 K 0 → +Vmax → –Vmax → +Vmax → –Vmax → 0 ±10 mV ΔV = 500 μV Vmax = ±3 mV ΔV = 100 μV 0.1 AP P 0.0 0 1 2 3 4 5 Applied V (mV) T = 295 K 6 dI/dV (μS) 3 0 –3 e P P ~350 s Power (nW) Current (nA) dI/dV (μS) d 200 100 0 –100 –200 –300 AP ~310 s 20 External magnetic field (Oe) c T = 295 K V = 5 mV TMR = –157% –20 170 –2000 –1000 TMR (%) We now utilize the case of the spinterface to illustrate several key considerations of how spintronic selectors and PM centers can interact to form the spin engine’s transport path. Upon connecting the spinterface to the PM state (see Fig. 1a), spinconserved quantum tunneling conditions the resulting spinpolarized landscape in the following signiﬁcant ways. First, the spinterface’s density of states (DOS) with a spatially oriented spin polarization generates a corresponding spintronic anisotropy in the PM state’s stochastic spin distribution27,28, thereby deforming the PM’s Bloch sphere of spin states. This generates an energy difference Δ between the PM center’s spin states and increases the probability that an electron tunnel onto/off of the PM if its spin is aligned to the spinterface’s spin referential (see Fig. 1b). The ensuing preferential charge ﬂow for that spin channel effectively shifts2 the spinterface/FM metal’s Fermi level by Δϕ toward that spin state of the PM center. We are thus describing how the spinterface can modify a metal’s properties29, namely its Fermi level position, through an additional mechanism. In a perfectly symmetric magnetic tunnel junction (MTJ) that implements spin-conserved tunneling between these two key ingredients—spinterfaces and a PM site—no net current (I) should ﬂow in the MTJ’s parallel (P) orientation of electrode magnetizations. However, in the MTJ’s antiparallel (AP) magnetic state (see Fig. 1a), the two FM electrode Fermi levels are shifted away from one another, each toward the corresponding spinterface-selected spin state of the PM center. The resulting spontaneous bias voltage ΔV between the FM electrodes thus scales with the amplitude of the spinterface’s spin polarization and the energy difference Δ between the PM center’s spin ↑ and ↓ states. Since an experimental MTJ cannot be exactly symmetric, one may also anticipate a spontaneous bias, or current, in the MTJ’s P state, albeit of lower amplitude. To generate work, the spin engine harvests energy from the spin ﬂuctuations that are thermally induced on the PM center. This thermal spin state mixing on the PM center enables current to ﬂow from one spinterface to the other, even against the built-in ΔV in the MTJ’s AP magnetic state. The spin engine thus requires that Δ ≤ kBT, and thus a balance between the tunneling-induced energy shift Δϕ of the spinterface state to the PM center’s spin state and thermal ﬂuctuations, as weighed by the spinterface’s spin polarization (see Fig. 1c). This thermal energy harvesting can be expected to cool the PM center. Furthermore, the fully spinpolarized current ﬂowing across the spinterface perturbs the FM ground state of the electrode through a spin accumulationinduced interfacial resistance30. The resulting heat generation in the FM electrode must be dissipated for our spin engine to work. Finally, the spinterface’s low density of highly spin-polarized states may be beneﬁcial to RT operation. Indeed, it protects the energetically discrete PM spin states against thermal broadening from the FM electrodes. As discussed theoretically in Supplementary Notes 1 and 2, the thermal ﬂuctuations in current are rectiﬁed ﬁrst upon transport from the FM electrode onto the spinterface, and furthermore upon transport from the spinterface onto the PM center’s spin state. This, along with the spinterface’s high spin polarization, strongly dampens any energy smearing of the PM center’s discrete spin states. The resulting energetically sharp, spin-polarized effective current path involving the striped DOS of the FM electrodes and spinterfaces is schematized in Fig. 1c by the yellow band. With the support of analytical and ab-initio theories, we believe to have observed an experimental realization of this spin engine through measurements across a single MgO MTJ at room temperature (RT; see junction statistics in Supplementary Note 3). As described hereafter, this MTJ integrates Co/C spinterfaces with nearly total spin polarization26, and paramagnetic C atoms31 on the oxygen vacancy sites of the MgO tunnel barrier. Referring to P 300 200 100 0 –100 –200 –10.0 AP ~0.25 mV 4 0 AP 2 3 Applied V (mV) T = 295 K kBT = 25.4 meV ~1.3 mV IP and IAP of opposite sign –7.5 –5.0 –2.5 0.0 2.5 5.0 7.5 10.0 Applied bias voltage (mV) Fig. 2 Spintronics-driven power generation at room temperature. a, b Magnetic ﬁeld dependence of current I (H) at a +10 mV and b +5 mV measured on the 20 μm-diameter magnetic tunnel junction (MTJ). c–e Bias dependence of c current and d numerically derived conductance dI/dV in the MTJ’s parallel (P; black; H = −2000 Oe) and antiparallel (AP; red; H = 250 Oe) states, and e the inferred tunneling magnetoresistance (TMR) bias dependence. Two sets of 0 → +Vmax → −Vmax → +Vmax → −Vmax → 0 sweeps spanning ± 10 mV (±3 mV) with 500 μV (100 μV) steps are shown. Discrete I(V) points obtained from the I(H) data of a and b are represented using semi-transparent crosses. TMR data <−100% (within the gray zone of c; see b) is achieved when IP and IAP are of opposite sign at constant V. A non-zero current that depends on the MTJ’s magnetic state is observed for V = 0. Despite the ~50 meV expected thermal smearing for this 295 K measurement, the +300% TMR peak has a full-width-at-halfmaximum of ~1.3 meV and statistically relevant conductance oscillations with a ~0.25 meV width are observed (see d inset). The inset to d shows the mean conductance over 4 bias voltage sweeps and the resulting standard deviation as error bars. This spectral sharpness is a direct signature of the spin engine. The inset to c shows how the bias-dependent output power depends on the MTJ’s magnetic state and can exceed 0.1 nW at room temperature Fig. 2a, we observe a negative tunneling magnetoresistance ratio, i.e. Since the external magnetic ﬁeld is static, and we do not expect a spin texture in our FM electrodes, a spin motive force explanation1,32,33 seems unlikely. In these and our experiments, no explicit temperature difference between electrodes, or temperature gradient, is applied to the device, such that a spin caloritronics34 explanation, while possible, is not obvious. We further discuss in the Methods and Supplementary Note 5 how photovoltage/photocurrent and conventional/spintronic thermovoltage17,35 artifacts can be excluded here. These I(H) datapoints are conﬁrmed through I(V) measurements at RT in the MTJ’s P and AP states (see Fig. 2c), which reveal the following features: (1) at V = 0, IP ≠ IAP ≠ 0, with an amplitude that also exceeds any experimental offset by nearly 2 orders of magnitude; (2) a non-zero applied bias V leading to a measured I = 0 whose amplitude depends on the MTJ’s magnetic state; (3) power generation above 0.1 nW whose bias dependence depends on the MTJ’s magnetic state, with a maximum current IAP ≈ −70 nA at V = +1.4 mV (see panel (c) inset using data from Supplementary Fig. 5a; (4) bias-driven oscillations in current that depend on the MTJ’s P/AP magnetic state, and thus on spin-dependent transport; (5) a bias range for which IP and IAP are of opposite sign, leading to TMR < −100%. These features of the 0 → +Vmax → −Vmax → +Vmax → −Vmax → 0 I(V) sweep are reproduced with high ﬁdelity in Fig. 2c for another such sweep with differing maximum applied bias Vmax and bias step (i.e. a differing effective bias sweep rate), as well as by additional datasets (see Supplementary Note 4). This eliminates any junction instability/memristive/Joule heating explanation36. (6) The numerically derived junction conductance dI/dV of the data of Fig. 2c, shown in Fig. 2d, reveals spintronically determined conductance jumps, and spectral features as low as 0.25 meV— despite the 2kBT ≈ 50 meV thermal smearing expected at 295 K— that are statistically beyond the error bar (see Fig. 2d inset and Supplementary Note 6 for full dataset and error bars) thanks to an excellent signal-to-noise ratio. This spectral sharpness is also witnessed through a 300% TMR peak with a full-width-half-max of ~1.3 meV (see Fig. 2e), which arises from a combination of local maxima(minima) in IP(IAP) at V = −3.5 mV. This non-optimized device’s spintronic performance at 295 K rivals the 600% record for FeCoB/MgO-class MTJs—obtained through a 20-fold performance increase over 7 years37,38 — but since the Co electrodes cannot be bcc-oriented here20,39 (see Methods), this spintronic performance cannot arise from symmetry ﬁltering. Within a simple Jullière model20 interpretation of the TMR amplitude, and from our analytical theory presented hereafter, this can be ascribed in part to the high transport spin polarization of the Co/C spinterface26 at RT. This spectral sharpness in magnetotransport features at RT despite the expected thermal broadening, and the excellent signalto-noise ratio, can be interpreted as an experimental signature of our spin engine at work. Indeed, according to quantum thermodynamics4,7, the harvesting of spin ﬂuctuations on the PM centers is expected to lower their temperature. From kBT ≈ 0.25 meV, we estimate an effective electronic temperature of the PM centers of 3 K. This cooling is the manifestation of harvesting energy from the PM centers’ spin ﬂuctuations upon spin rectiﬁcation in the junction. Since a non-zero current is present at V = 0 across this normally passive component, we observe that the MTJ is intrinsically out of equilibrium. Consequently, to further link our experimental results with our conceptual spin engine, we analytically consider an out-of-equilibrium nanotransport path across the MTJ comprising two PM centers (see Fig. 3a and the Methods/ Supplementary Note 1 for details). Their initially discrete energy I(nA) ARTICLE 10 Bias voltage (mV) Fig. 3 Analytical theory linking experiments to the spin engine. a Schematic of the analytical model of spin-conserved transport across a paramagnetic dimer (PM1 & PM2 centers) separated from each ferromagnetic (FM) lead by a spinterface. The calculated case of the P magnetic state at V = 0 is shown. b Theoretical bias dependence of spin ↑ (solid) and spin ↓ (semitransparent) current in the magnetic tunnel junction’s parallel (P; black) and antiparallel (AP; red) magnetic states. c Theoretical and d averaged experimental bias dependencies of current in the MTJ’s P (black) and AP (red) magnetic states, and of the resulting tunneling magnetoresistance (TMR; green), using the same current/TMR scales. Experimental standard deviations are shown as error bars in orange. Within an apparent factor in the voltage scale, the analytical model including the out-of-equilibrium hypothesis strongly mimicks the room-temperature (RT) experiment, thereby linking it to the spin engine levels (gray lines) are broadened to form a PM dimer as bonding/ anti-bonding and spin degeneracies are lifted (see PM 1&2 of Fig. 3a) through a magnetic exchange coupling that is biasdependent28,40. To place the junction out of equilibrium, we impose a spin splitting of the FM electrodes’ chemical potential. Following our spin engine proposal, our analytical model’s magnetic interactions (with Heisenberg, Ising and Dzyaloshinskii–Moriya contributions) result in a spintronic anisotropy27,28 onto PM1 and PM2 due to the spatially orientated, spin-polarized DOS of each FM electrode, as mediated by spinterfaces (SP1 & SP2). Consistently with our experimental results, we constrain the model’s 7-fold ﬁtting parameter space using the following physical requirements: 1) the parameters should realistically describe the MTJ’s outer properties (FM electrode + spinterface), including a higher spin polarization at the lower Co/MgO MTJ interface due to C dusting26 (see Methods); 2) these outer properties COMMUNICATIONS PHYSICS \| (2019)2:116 \| https://doi.org/10.1038/s42005-019-0207-8 \| www.nature.com/commsphys ARTICLE COMMUNICATIONS PHYSICS \| https://doi.org/10.1038/s42005-019-0207-8 DOS (state/energy) 40 30 DOS (state/energy) should remain identical in the MTJ’s P/AP magnetic states; 3) only minor changes to the PM dimer’s properties are allowed between the P and AP cases. To account for T = 295 K, the FM electrodes’ Fermi level is broadened by 26 meV (not shown in Fig. 3a). Results of this analytical model shown in Fig. 3b, c were acquired using a same set of parameters (see Methods for details) for the MTJ’s P/AP outer properties: sp = 8.4, SP = 2, pL = pR = 0.35, AP = 0.3, E0 = 0, with pR and AP changing sign upon P ↔ AP, while we introduced minor variations in the PM dimer’s starting conditions for P(AP): e0 = −2.5(+0.85) and ASYM = 0.5 (−0.75). Parameters are described in the Methods. Supplementary Fig. 2 shows the complex bias dependence of this spintronic potential landscape for each spinterface/PM center (SP1, PM1, PM2 & SP2), depending on the spin channel and the MTJ’s P or AP magnetic state considered. Note how our model fulﬁlls the spin engine’s Δ < kBT condition. Referring to Fig. 3b, we observe a bias anti-symmetric imbalance in the oppositely propagating spin channels of current, which strongly depends on the MTJ’s P/AP magnetic state. This leads to a sizeable spintronic difference in current, in particular at V = 0. We recopy the IP(V), IAP(V) and TMR(V) experimental data of Fig. 2c/e as Fig. 3d in order to compare them with their analytical counterparts, shown in Fig. 3c. Despite a skewed bias position that could underscore the simplicity of the bias voltage distribution (see Fig. 3a), our out-of-equilibrium analytical model reproduces all trends and salient features of the experimental magnetotransport data. This includes the spintronically dependent non-zero current at V = 0, large TMR peak at V < 0 and the bias region for V > 0 with differing signs of IP and IAP. A degraded agreement at large V likely reﬂects how our model only considers sequential transport across the 4 QDs, and not direct transport between the FM electrodes, which can become signiﬁcant as the QD levels are energetically shifted away from one another. This agreement between theory and experiment compares quite favorably with respect to the state of the art21,22,41,42. In general, compared to low-temperature transport across wellcharacterized quantum objects (e.g. from single atoms and dimers to molecules and atomic clusters43–49) thanks to a scanning tunneling microscope (STM), it is thus far difﬁcult to assemble and ascertain the effective nanotransport path50 in a solid state device, especially for the oxides used as MTJ barriers. Here, uncontrolled imperfections such as oxygen vacancies in the MgO tunnel barrier can concentrate electronic tunneling transport across a macrojunction onto a nanotransport path50,51, such that the device operates due to a rare tunneling event52. This is what enables53 the spin transfer torque effect underscoring key MTJbased technologies54,55. As detailed in Supplementary Notes 7 and 8, while descriptions of the PM dimer in terms of Mn atoms or oxygen vacancies are much less likely here, paramagnetic C atoms occupying oxygen vacancy sites in MgO are possible considering our MTJ stack with C-dusted MgO interfaces. Indeed, carbon capture by single/double oxygen vacancies, which are present in our MgO21,23,53, is energetically favorable (see Supplementary Note 7 and the work of Tiusan et al.56) and can yield both paramagnetic monomers (see Supplementary Note 7) and dimers31. Our ab-initio theory shows that the C–C distance is crucial in order to reproduce our analytical model’s results: only in a 4th nearest-neighbor positioning does the C dimer simultaneously exhibit AF coupling (favorable over FM by 0.125 eV, i.e. above experimental kBT) and generate four states around the Fermi level EF of a Co/MgO/Co MTJ (see Fig. 4 and Supplementary Note 9). On the other hand, C pairs in 1st, 2nd, 3rd, and 5th nearestneighbor conﬁguration generate a FM state (see Supplementary Note 7), which would be inconsistent with our analytical model. This stringent C impurity positional requirement on the oxygen 20 10 2kBT 5 0 –5 –10 10 –0.2 –0.1 MgO valence band 0 0.0 0.1 0.2 E –EF (eV) –10 MgO conduction band C 2p states –20 Spin ↑ Spin ↓ –30 –40 –5 –4 –3 –2 –1 0 1 E –EF (eV) 2 3 4 5 6 Fig. 4 Origin of the experiment’s paramagnetic centers. Ab-initio calculations of the spin-resolved density of states (DOS) of MgO containing a carbon dimer in 4th nearest-neighbor conﬁguration. The paramagnetic, antiferromagnetically coupled C dimer generates energy levels around the magnetic tunnel junction’s Fermi level EF, including four spin-degenerate states that intersect EF, thereby reproducing the analytical model’s description of PM centers 1 and 2. See Supplementary Note 9 for the determination of the MTJ’s EF energy position sublattice might explain why our spin engine was experimentally observed only once out of ~200 attempts (see Supplementary Note 3). Discussion To achieve routine experimental reproducibility, we propose that all spintronic selector tracks be attempted (see introduction), noting that, in addition to the two published reports1,2, similar effects were observed at low temperature on MTJs with manganite half metals57. In all cases, control over the spatial position and density of the barrier’s PM centers will be required with a precision that, at this time, remains the domain of model STMassembled junctions43–49. Considering that all reports involved microscale devices, this suggests reducing the junction’s lateral size from the micro- to the nano-scale. In an oxide track, one may study tunnel barriers in which an oxygen vacancy-rich central region—achieved e.g. by varying oxygen concentration in an Ar sputtering plasma during growth51, is nominally seeded with impurities to be trapped by these vacancies as PM centers. Control over the electronic properties of, and magnetic interactions58 between, PM centers in molecules suggests another, organic-based track using spintronic nanojunctions59,60. Whatever the route, except for an AF-coupled PM dimer, the PM center(s) should experience dominant tunneling from one spintronic selector in order to adopt that selector’s spin referential (see Fig. 1b and discussion). This can arise by tuning the selector/ PM center tunneling rate through the insertion of an oxide/ organic interlayer. Cleverly crafted operando techniques50 that can directly characterize the PM center’s properties within the device’s nanotransport path can boost research efﬁciency. Overall, MgO spintronics represents a compelling route. Indeed, it beneﬁts from both industrial penetration54,61 and knowledge on how oxygen vacancies craft the spintronic nanotransport path21–24,53, boasts lateral sizes down to 4.3 nm62, and has been conjugated with half-metallic electrodes operating at RT17. PM centers can be formed in MgO by trapping C, N or Si on oxygen vacancies (see Fig. 4, the work of Wu et al.31 and Supplementary Note 7). To complement this heat description of our work, we brieﬂy note in the Supplementary Note 10 that the rectiﬁcation of COMMUNICATIONS PHYSICS \| (2019)2:116 \| https://doi.org/10.1038/s42005-019-0207-8 \| www.nature.com/commsphys 5 ARTICLE COMMUNICATIONS PHYSICS \| https://doi.org/10.1038/s42005-019-0207-8 thermal ﬂuctuations, which are present experimentally and analytically, can generate work when combined with an increase in the entropy of quantum information11 arising from spin transport onto the PM center due to its spin ﬂuctuations. A quantum thermodynamical theory along a similar spintronic path has been proposed8, while classical electronic implementations using capacitively coupled quantum dots have been demonstrated at low temperature3,5,63. Our results should thus generate research initiatives on quantum electronic circuits at the rather unexplored intersection between quantum thermodynamics3–12 and spintronics20,34,54. More generally, our work also indicates that the high transport spin polarization, and low density of states, of spinterfaces represent a compelling approach to integrating the quantum properties of nano-objects within a solid-state device’s operation at room temperature, beyond proof-of-concept electronic decoupling strategies43,64. The MTJ used to demonstrate our spin engine is an industrial-grade microelectronic device class used as the read head of hard disk drives, and with promising potential toward low-power information storage54 and bio-inspired computing55. If routine reproducibility can be achieved, then for a typical STT-MRAM61 2D array of 50 nm-wide MTJs with a pitch of 90 nm, assuming that the 0.1nW occurred through a single conduction channel, the resulting power density would be substantial. As an example, a suitable series/parallel assembly on a 1 cm2 chip could deliver ~29.4 W of power to a 0.1 Ω load at 3.5 V. See Supplementary Note 11 for more details. Even considering 1% efﬁciency due to engineering issues (e.g. managing heat ﬂow, interconnect resistances…), this power density would still exceed the raw solar power density on the earth surface by a factor of three. Further research may see the MTJ play a key role not only in information & communication, but also renewable energy, technologies. that are centered around EF at V = 0 and are spin-split by SP. A constant E0 = 0 was used. PM 1&2 model the paramagnetic dimer as two spin states S1 and S2 that, initially, are energetically discrete, are positioned eo away from EF at V = 0, and are energy-split by ASYM but are not spin-split. Current ﬂows between the FM electrodes across SP1/PM1/PM2/SP2 through a tunneling rate T, which was ﬁxed at [1 1 1] between SP1/PM1; PM1/PM2; PM2/SP2. Finally, pL(pR) describes a possible spin polarization of the tunelling transmission between the left(right) FM lead and SP1(SP2). The MTJ’s AP state is described by switching the sign of pR and AP, i.e. by ﬂipping the right-hand FM electrode magnetization. This experimentally corresponds to the free Co layer of the top FM electrode. AP = 0.3 is consistent with an experimentally larger spin polarization of the C-dusted Co lower FM electrode26, and to d1 ≠ d2. Supplementary Note 1 further details the model’s transport formalism. Methods 2. Device preparation and batch characterization. Ta(5)/Co(10)/IrMn(7.5)/Co (4)/C(d1 = 0, 0.3, 0.6, 0.9)/MgO(2.5 nm)/C(d2 = 0, 0.3, 0.6)/Co(10)/Pt(4) samples (all thicknesses in nm) were sputter-grown on Corning 1737 glass substrates65. Stacks were post-annealed in an in-plane magnetic ﬁeld of 200 Oe for 1 h at a temperature Ta of 200 °C to magnetically pin the lower electrode thanks to the IrMn antiferromagnetic layer. This low annealing temperature precludes the diffusion of Mn into the barrier66, though it can promote C diffusion into MgO56. Samples were then processed by optical lithography67 into 20 μm-diameter MTJs, and measured on a variable-temperature magnetotransport bench. Within the study, 216 MTJs that exhibited neither a short-circuit nor an open circuit had a median R = 4395 Ω in the MTJ’s P state, i.e. a median R.A. product of 1.38E6 Ω.μm2. Only 1 out of 168 20 μm-diameter MTJs tested at T = 295 K, with d1 = 0.9 and d2 = 0, a ten-fold larger RA product (R = 57.7 kΩ, R.A. = 1.81E7 Ω.μm2; standard contact resistances) and anomalous TMR < 0 at V = +10 mV, revealed these peculiar power generation features and high TMR. An additional 48 MTJs did not exhibit I ≠ 0 at V = 0. We presume that annealinginduced C migration into MgO generated the MTJ’s spinterfaces and PM centers. The lower R found in passive MTJs suggests the presence of several conduction channels, each with a slightly different nanotransport path, such that electrical generation effects are averaged out. Possible transport artifacts. Measurements on this MTJ were conducted over 90 min in 4-point measurement mode within a dark cryostat that remained between 295.3 K and 294.5 K with sample heater off. Furthermore, these peculiar transport features strongly depend on the MTJ’s P/AP magnetic state. We in particular observe that I ≠ 0 at V = 0. We can therefore exclude thermovoltage photovoltage/ photocurrent, as well as any conventional or spintronic thermovoltage explanations. We discuss these discarded artifact sources in more detail in Supplementary Note 5. Analytical theory. The analytical model28,40 symmetrically segments the MTJ’s bias drop into 6 zones around the junction mid-point, for which V ≡ 0, using the same bias sign convention as in experiment. Current ﬂow across the left-hand (right-hand) FM electrode is modeled by a spin-splitting sp(sp × AP) of its chemical potential. The DOS of the spinterfaces SP1 & SP2 consists in 10meV-wide bands 6 Ab-initio theory. Within density functional theory, the electronic properties of the C dimer within MgO were computed using 64-atom supercells with a simple cubic structure with two substitutional carbon atoms in various conﬁgurations (see Supplementary Note 7 and Supplementary Fig. 8). These calculations were done using VASP code68 based on the projector augmented wave (PAW) method69 and the Pedrew, Burke, Enzerhof (PBE)70 generalized gradient approximation for the exchange-correlation potential. The kinetic energy cutoff value of 500 eV for the plane wave basis set and the convergence criterion for the total energy of 10−8 eV is used. The carbon-doped structures are fully relaxed using a conjugate-gradient algorithm, such that the forces acting on atoms be less than 0.001 eV/Å. A k-point mesh of 6 × 6 × 6 with the Methfessel-Paxton method with a smearing τ = 0.1 eV is used. See Supplementary Note 9 for the determination of EF within a Co/MgO(12 ML, i.e. ~2.5 nm)/Co MTJ. Data availability The data that support the ﬁndings of this study are available from the corresponding author upon request. Received: 3 May 2019 Accepted: 1 August 2019 Published online: 25 September 2019 References 1. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. Hai, P. 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Effect of high annealing temperature on giant tunnel magnetoresistance ratio of CoFeB∕MgO∕CoFeB magnetic tunnel junctions. Appl. Phys. Lett. 89, 232510 (2006). 67. Halley, D. et al. Electrical switching in Fe∕Cr∕MgO∕Fe magnetic tunnel junctions. Appl. Phys. Lett. 92, 212115 (2008). 68. Kresse, G. & Furthmüller, J. Efﬁcient iterative schemes for ab initio totalenergy calculations using a plane-wave basis set. Phys. Rev. B 54, 11169–11186 (1996). 69. Kresse, G. & Joubert, D. From ultrasoft pseudopotentials to the projector augmented-wave method. Phys. Rev. B 59, 1758–1775 (1999). 70. Perdew, J. P., Burke, K. & Ernzerhof, M. Generalized gradient approximation made simple. Phys. Rev. Lett. 77, 3865–3868 (1996). Acknowledgements We are grateful to H. Thierschmann, K. McKenna and J. Blumberger for useful discussions, and to Y. Henry for carefully reading our manuscript. Devices were synthesized at the STNano technological platform. We acknowledge ﬁnancial support from the Institut Carnot MICA (project ‘Spinterface’), from the ANR (ANR-09-JCJC-0137, ANR-14-CE26-0009-01), the Labex NIE “Symmix” (ANR-11-LABX-0058 NIE) and Vetenskapsrådet. This work was performed using HPC resources from the Strasbourg Mesocenter and from the GENCICINES Grant 2016-gem1100. Open access funding provided by Uppsala University. Author contributions M.B., E.B. and S.B. conceived the initial experiment. M.H. grew the sample stack. E.U., F.S., K.K. and J.A. implemented technological processing to make MTJs. E.U., F.S. and K.K. performed magnetotransport measurements. M.B., K.K., S.B., D.L., E.B., M.A. and W.W. analyzed magnetotransport results. B.V. and D.S. performed auxiliary measurements. J.F. designed the analytical models with input from M.B. K.K. applied the analytical models, with input from M.B. and J.F. B.T. COMMUNICATIONS PHYSICS \| (2019)2:116 \| https://doi.org/10.1038/s42005-019-0207-8 \| www.nature.com/commsphys 7 ARTICLE COMMUNICATIONS PHYSICS \| https://doi.org/10.1038/s42005-019-0207-8 and M.A. implemented DFT calculations with input from M.B. M.B., K.K. and B.T. prepared the manuscript. All authors commented on the manuscript.	19,054
2018PSLEH006_8	French-Science-Pile	Open Science	Various open science	2,018	Transferts Privés de Revenus et Développement : trois Essais Appliqués à l'Amérique Latine	None	English	Spoken	7,468	10,161	Cash transfers to the elderly might impact children’s nutrition and health by allowing households to spend additional income on more nutritious foods and health care, and by inducing changes in the way intra-household resources are allocated. The identification of these effects represents an empirical challenge because households with age-eligible members, might be wealthier, better endowed with genetics and have more appropriate nutrition habits and health practices. I address this issue by implementing a regression discontinuity analysis that exploits the fact that program eligibility exhibits a discontinuity at the age of 65 years old. Households where the oldest member is 64 y/o and those where the oldest member is 65 y/o are assumed to be almost identical except for the fact that the last are eligible to receive a Pensión 65 subsidy. Note that this strategy is very similar to the one followed by Galiani and Gertler (2016). However, while I use a discontinuity rule based on age eligibility, their design is based on the household poverty index. Although, this variable is also a determinant factor of household eligibility, I consider their choice more noisy because this same index is used by the Peruvian government to assign many other social programs. A large number of studies have assessed the effects of old-age cash transfers on poverty4 , resource allocation5 , living arrangements6 , labor supply 7 , household technical efficiency8 , mi4 See Barrientos (2005) for a discussion. For instance Case et al. (1996), Maitra and Ray (2003), Martinez (2009) and Galiani et al. (2016). 6 Reference s include Edmonds et al. (2005), Manacorda and Moretti (2005) and de Oliveira and Kassouf (2012). 7 For example Bertrand et al. (2003), de Carvalho Filho (2008), Hosegood et al. (2009), Galiani et al. (2016) and de Oliveira and Kassouf (2012). , users health10 and children outcomes like nutrition, health11 , child labor12 and schooling13 . Most have found, not only that these programs are effective in tackling poverty and improving the quality of life of the elderly, but also that they have important spillover effects within the household. In the specific case of spending and consumption patterns and its relationship with children nutrition and health outcomes, existing research generally studies both issues separately. In addition to Galiani and Gertler (2016), five other papers stand out for an empirical analysis of the relationship between elderly cash transfers and household spending/consumption. In a preeminent article about the expansion of the South African pension system in the early 1990’s, Case et al. (1996) analyze its effects on household spending patterns. The authors find positive effects on food and schooling, but no effect on health. In addition, arguing that beneficiary households are predominantly poor and that in South Africa the fraction of children living with a pensioner is quite high, they claim that these results are evidence of the re-distributive effectiveness of the program. Also in the context of the South African pension reform, Maitra and Ray (2003) analyze changes in the composition of household spending. Unlike Case et al. (1996), they find no impact in any of the 11 categories analyzed, including food, health and education. Furthermore, their results question the re-distributive effects of the program by showing that the amount of pension received decreases with the number of children in the household. Fan (2010) studies the effects of an important pension reform in the mid 90s in Taiwan, that introduced a monthly pension of NT$3, 000 to adults over 65 years old not receiving pensions from social insurance. He finds that a 1 NT dollar granted raises household consumption by 41 cents. In the Latin American context, besides Galiani and Gertler (2016), there are, to my knowledge, only two studies of this type. Martinez (2009) presents an evaluation of the Bono Solidario BONOSOL, an old-age cash program in Bolivia very similar to Pensión 65. The author finds a positive effect on food consumption, equivalent to 97% of the program annuity, for the whole population sample, and 165%, for the sub-sample of poor rural households. Households also increase spending on medical services like doctor visits and medicines. Finally, Galiani et al. (2016) evaluate the Mexican program Adultos Mayores, a cash transfer scheme targeting to rural adults over 70 years old. They find that the program is associated 9 For instance Posel et al. (2006) . See Atalay and Barrett (2012) and Galiani and Gertler (2016). 11 For example Duflo (2000, 2003), Case (2001) and Case and Menendez (2007). 12 See Edmonds (2006), de Oliveira and Kassouf (2012) and de Carvalho Filho (2012) among others. 13 References include Hamoudi et al. (2005) Edmonds (2006), de Carvalho Filho (2012) and Gutierrez et al. (2015). Introduction with an increase in household consumption of 23%. Authors affirm that, as, almost without exception, adults over 70 years in rural Mexico live with another relative, this rise in household consumption is evidence that beneficiaries share the transfers with their families. As it was mentioned before, none of these articles directly evaluates the program effects on children’s nutrition and health. A reasonable number of studies have addressed this question in the case of South Africa. The most influential is, perhaps, the work of Duflo (2000, 2003), where she evaluates the impact of this reform on grandchildren’s weight for height and height for age. She finds that pensions received by women have a positive impact on girls under 5 years old. Point estimates are 1.16 standard deviations on height for age and 1.19 on weight for height, two years after the reform. In contrast, no effect is found on boys and for pensions received by men. Case (2001) reaches similar results, although she uses a different dataset and another identification strategy. In this case, however, positive health outcomes extend also to co-resident adults and boys and are not exclusively driven by elderly women. As an added value, this study uses qualitative information14 to investigate the mechanisms through which pension income fosters health outcomes. The analysis suggests that these households have better sanitation facilities and are less likely to skip meals and reduce the size of their meals. This conclusion is confirmed by Case and Menendez (2007) in an evaluation of pension beneficiaries and their co-resident prime-aged adults and children. Using a survey applied to a very specific sample of 29015 households in the rural sub-district of Limpopo Province (South Africa), the authors show that the presence of a pensioner improves child food security, by diminishing the probability of skipping meals due to income constraints. The analysis presented in this chapter adds to this literature in two ways. First, unlike earlier similar studies, it attempts to explicitly map the relationship between cash transfers targeted to the elderly, household spending patterns and the nutrition and health status of very young children. Given that no data are available to address both questions simultaneously, I separate the analysis in two parts. I start by investigating the relationship of Pensión 65 with the monetary spending of households with children under 5 years old, using the 2015 round of the national household survey, Encuesta Nacional de Hogares sobre Condiciones de Vida y Pobreza (ENAHO). 14 Interviews to pensioners about their perception and behavior after receiving retirement payments, such as: “What did you start doing differently when you received your pension?” and “In what ways did your life become better when your pension started, if any?”, among others. 15 This design included 124 households with and 166 without at least one age-eligible older adult. 138 Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru Then, I examine the association of the program on the nutritional and health status of this population (children under 5 y/o), based on the 2015 demographic and health survey, Encuesta Demográfica y de Salud Familiar (ENDES). This way I can provide some understanding of the transmission mechanisms from cash transfers to the elderly towards the improvement of children’s health. More precisely, I directly test the specific role of changes in nutrition and health spending, a channel frequently mentioned, but little studied, in the existing literature on the subject. The second contribution is related to the fact that the empirical literature documenting the effects of old-age transfers on children’s health is almost exclusively about the South African pension reform. A case which is very difficult to extrapolate. This analysis will, therefore, help to improve the external validity of this literature, applying a very similar strategy to a completely different context. Results suggest that Pensión 65 eligible households with young children increase monetary spending by 75% of the value of the transfer. This additional income shows up in more purchases of vegetables and grains (legumes) and an increase of health expenses. In parallel, coresident children of these ages show significantly better nutrition and health outcomes, such as: weight for age (1.47 standard deviations higher), weight for height (1.35 sd higher), height for age (1 sd higher) and body mass index (1.15 sd higher). In addition, their probability of suffering from underweight is 39 percentage points less and their likelihood of mild anemia 35. In line with previous research on the re-distributive effects of subsidies to the elderly in developing countries16 and with the evaluations of other cash transfer programs recently implemented in Peru17 , these findings tend to support the hypothesis that income constraints play a key role in determining the well-being of poor households with very young children. In addition, they constitute new evidence that households do not function as unitary entities18 , and that old-age adults have major decision-making abilities and play a key role in channeling investment towards young children. The rest of the chapter is organized as follows. Section 3.2 describes the setting of Pensión 65, presenting the context and the main characteristics of the program. Section 3.3 outlines the data and provides some descriptive statistics of the samples. Section 3.4 presents the empiri16 See Martinez (2009) and Duflo (2000, 2003) among others. For instance Perova and Vakkis (2009). 18 Reference on non-unitary household models includes Bourguignon et al. (1993), Browning and Chiappori (1998) and Blundell et al. (2005). 17 3.2 Background 139 cal strategy, a regression discontinuity design based on age eligibility. Section 3.5 reports the main findings, presents some sensitivity and validity tests and discusses potential confounding factors. Section 3.6 concludes. 3.2 Background This section presents some key information to better contextualize Pensión 65. It first outlines the main traits of Peru’s elderly population and then presents the characteristics of the program. 3.2.1 Peru’s Elderly Population The elderly population in Peru has increased by 65% in the last 15 years, whereas total population has grown 20% during the same period. By 2015, 7% of the total population was 65 or older, numbering more than two million individuals, and 2 in 5 households had at least one adult in this age group. Estimates by the Instituto Nacional de Estadística INEI predict that this population will reach 9 million by 2050.19 Table 3.1 summarizes some basic statistics describing with more details this population , extracted from the 2015 of the ENAHO . These figures indicate that in Peru, most of the 65-yearsold-and-plus adults live in urban areas (80%), with more than one third concentrated in Lima. Elderly households are composed, on average, of 3 members: only 0.35 children under 5 y/o and almost 2 adults over 55 y/o (counting herself). These old-age adults are, on average, 74 years old and, due to the widening sex gap in terms of life expectancy in favor of women, more than half are females. Very worrying is the fact that 40% of these households live in extreme poverty, whereas the same figure is 31% at the national level. This is evidence that for Peru, as it is the case of most developing countries, poverty is a big concern when it comes to the elderly population.20 Although older people are generally less likely to participate in the labor force, about 45% of the individuals of this sub-sample are employed, most of them in the informal sector.21 Similarly, 6 in 10 do not have access to formal health insurance, meaning that a big part of them is not protected against health risks, which tends to be very high at these age levels. With regards to pension coverage, only 36% affirm to be affiliated to a contributory regime, with half of them 19 See INEI (2015). The definition of extreme poverty used for these calculations, and throughout this entire article, draws on the one employed by the Peruvian household targeting system to evaluate welfare and provide social benefits, i.e. the Sistema de Focalización de Hogares (SISFOH). This definition is explained in detail in the next section. 21 This rate is 70% among the 15-65-years-old. Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru claiming been actually receiving pension income (24%). The average amount received in the last year was 790 Soles, slightly less than the monthly minimum wage in Peru, which was 850 in 2015. Peru’s contributory pension system is an employment based regime, in which only individuals with a stable formal job take part. In consequence, there is a direct association between pension coverage and labor informality. By 2015, Peru was among the first countries in the region in terms of informality, with 73% of total employment in this sector.22 3.2.2 Pensión 65 General Characteristics In order o address, at the same time, low pension coverage and old-age poverty issues, in 2011, the government of Peru launched an initiative called Programa Nacional de Asistencia Solidaria Pensión 65, a cash transfer program, addressed to extremely poor elders (over 65 years old) with no access to the contributory regime.23 This program was inspired by the non-contributory pension schemes created in other countries of the region since the beginning of the nineties, like the Benefício de Prestação Continuada (Brazil, 1996), The Bono Solidario Bonosol (Bolivia, 1997), the Pensión Básica Solidaria de Vejez (Chile, 2008) and the Programa Colombia Mayor (Colombia, 2013).24 The Pensión 65 allocation was fixed at 125 Soles per month (24 US Dollars), representing 23% of national expenditure per capita, and included bi-monthly payments received through the agencies of the Nacional Bank (Banco de la Nación), for beneficiaries living close to urban centers, or through securities transportation companies, for the population living in more remote areas. Even though the subsidy does not have any conditionality associated, it also included two complementary actions. The first was granting beneficiaries with free access to health care services and implementing a specific health agenda targeted to the elderly. The second was a strategy to integrate more the elders in the community, through the recognition of their ancestral knowledge.25 22 Author’s calculation using the 2015 ENAHO. A pilot version (Bono Gratitud) was launched in July 2010 covering 21,783 beneficiaries in the departments of Apurímac, Ayacucho, Callao, Huancavelica and the district of Lima. 24 For a more comprehensive list see the Base de datos de programas de protección social no contributiva en América Latina y el Caribe published by the ECLAC and available in http : //dds.cepal.org/bdps/ 25 Some examples include the publication Vidas, a book made with the objective of visualizing this population, the traveling photographic exhibition Ciudadanos and the national contest Los abuelos ahora, which seeks that primary school children perform works where the protagonists are the elderly. 23 3.2.2 Pensión Beneficiary Selection Process Beneficiaries are identified by program officials and recorded in a list published every two months at the district level. To qualify for the program, individuals need to fulfill the following criteria: (i) being 65 years old and more, (ii) being living in extreme poverty conditions, (iii) having a valid ID, (iv) not being receiving pension income and (v) not being benefiting from any government social program, other than health insurance SIS, literacy programs, food assistance programs, the national program of direct support to the poorest Juntos or any program part of the comprehensive plan for reparations PIR. The identification of 65 years old individuals living in extremely poor households is made accordingly to a welfare evaluation and classification system known as SISFOH, that is used to identify potential beneficiaries of different social programs and government subsidies in Peru. It is based on a general household census (Padrón General del Hogares PGH), built from three different sources: (i) administrative records of income earners of the formal sector of the economy, (ii) the censuses of households living in poverty (Bolsones de pobreza), refreshed every three years in urban areas and four years in rural areas, and (iii) system of household registration on demand. The classification process, represented graphically in Figure 3.1, uses three different filters: (i) labor income, (ii) a composite household index of quality of life and (iii) water and electricity consumption. When at least one household member works in the formal sector, the first identification criteria is labor income. If household monthly labor income is greater than 1,500 Soles (570 US Dollars) the household is automatically classified as not poor. On the contrary, if the household is below 1,500, a quality of life index, the Índice de Focalización de Hogares IFH, is evaluated. The IFH index is built using a set of variables weighted according to three geographic areas: Metropolitan Lima, other urban areas and all rural areas (Table 3.2).26 These weights are used to build a raw index P Ihj for each household h in cluster j.27 Considering, for example, a household in Lima, cooking with carbon, using water from a pipe and living in a house made of wood, the first three addends of this index would be -0.33, -0.41 and -0.48. Then, the following equation, Equation (3.1), was applied, in order to standardized the 26 The selection of these variables and the calculation of the weights was done by the Peruvian Ministry of Finance using data from the 2009 ENAHO and includes two steps. The first was to identify the correlations between a set of candidate explanatory variables and poverty (based on a Sommers test). The second was to implement a Principal Component Analysis to select the items explaining more variation. 27 Clusters were built by program authorities to group homogeneous geographic zones (by department, region and domain) in terms of their own poverty lines. The 15 clusters are presented in Table 3.3. Cluster 2, for instance, includes the rural and sylvan areas of the departments of Ayacucho, Junin, Loreto, Puno, San Martin and Ucayali and the rural, northern highlands of the departments of Cajamarca and Lambayeque. 142 Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru values of the index between 0 and 100: P I0hj = 100 ⇤ P Ihj max P Ihj min P Ihj min P Ihj (3.1) Extreme poverty is determined by comparing the standardized index of each household with the corresponding cluster threshold (Table 3.3). Households with an index below or equal to the threshold were classified as eligible. If, on the contrary, the IFH value was above the threshold, eligibility was decided on the basis of water and electricity consumption. A household was then eligible if water and electricity expenditures were below 20 and 25 Soles per month, respectively.28 Finally, if no member was working in the formal sector, poverty classification was based only on the value of the IFH index and water and electricity spending, or, only IFH if no information on the last criteria was available. This process produces a first list of potential beneficiaries sent to district officials in order to verify that they were alive and effectively eligible (in terms of poverty). in addition, these district authorities had the mandate to ask them for a sworn statement of eligibility and an ID copy. Back in Lima, program officials implemented a very last verification crossing these lists with other administrative databases like the ones produced by the Social Security Agency (ONP), the Superintendency of Banking and Insurance (SBS), the Registry of Identification and Marital Status and the health insurance system (ESSALUD). Program Coverage In the first months of operation, Pensión 65 prioritized districts with extreme poverty incidence greater than 50 % in 13 departments, covering only 377 out of 1,840 districts and 31,143 beneficiaries (corresponding to 11% of the eligible population29 ). However, the program expanded rapidly, reaching national coverage at the end of 2013 (Figure 3.2). Calculations based on the number of beneficiaries, 501,681 individuals (MIDIS, 2016), and the total eligible population, calculated from the ENAHO (783,876), reveal that 64% of the eligible population was receiving a Pensión 65 grant, by 2015. It should be noted that this figure differs from the rate of 72% given by the government, that estimates the eligible population at 688,301 individuals for the same year 2015 (DGPP-MEF, 2015). The inconsistency, recognized by program officials (DGPP-MEF, 2015), is mainly due to an under-coverage problem of the general household census (Padrón General del Hogares PGH) 28 29 These values correspond to 7.6 and 9.5 US Dollars. According to program official reports, the eligible population was estimated at 280,976 in 2011. 3.3 Data and Descriptive statistics 143 used as the basis for the identification of potential users.30 . Since the PGH is one of the inputs used to target beneficiaries, this is also the main reason behind the low coverage rates of the program. Table 3.4 and Figure 3.3 present actual program coverage, for the 65-years-old-and-plus population, according to the 2015 ENAHO. 3 in 10 adults over this age meet eligibility requirements to benefit from the program: 41% live in extremely poor households, 99% have an ID, 94% do not receive pension income and 94% do not receive any other subsidy. On average, participants affirm being benefiting from the program for the last 18 months, with the first 44% getting the allocation before January 2014 (Figure 3.3). Beneficiaries received, on average, 1,320 Soles from the program during the last year, more than two times the annual pension income of an average 65 and plus individual. 3.3 Data and Descriptive statistics The following analysis uses data from two nationally representative surveys, collected by the National Statistical Institute of Peru (INEI). The first one, the 2015 round of the ENAHO, is used to investigate the relationship between Pensión 65 and the spending paths of households with children under 5 years old; while the second one, the 2015 ENDES, is required to examine the association between the program and the nutritional and health status of this population. 3.3.1 Households with Children under 5 y/o Household data from the ENAHO was collected between the months of January and December of 2015. It gathers information on household characteristics, labor participation, income, investment, consumption and, of particular relevance for this paper, expenditures, where households report all the products, goods and services purchased during the last year.31 As the main objective is to evaluate whether Pensión 65 contributes to enhance the nutrition and health of young children, I focus on the sub-sample of households with members under 5 years old. In addition, given the characteristics of the program and the econometric strategy, I restrict the analysis to extremely poor households32 , with at least one eligible adult (i.e. holding a National Identity Document, not receiving a pension and not receive any subsidy or 30 According to the Ministry of development and social inclusion, by the end of 2014, the PGH only included information about 63% of the population of Peru (DGSE-MIDIS, 2016) 31 In the original survey, depending on the items, there are different reference periods. For instance, food spending is reported for the last 15 days, housing and personal care goods and services for the last month, clothing and footwear for the last 3 months, durables and education expenditures for the last year and so on. In order to allow comparisons, the INEI, inputs, deflects and converts all values and quantities in annual measures. 32 Extremely poor households were identified following the same process used by program officials to built the list of potential Pensión 65 beneficiaries (Subsection 3.2.2). 144 Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru government grant), having between 55 and 75 years old (i.e. 119 months bandwidth).33 This leaves a final sample of 963 households (Table 3.5). Outcomes of interest include: total spending broken down into 5 broad components (food, education34 , health services, durables35 and housing36 ), 12 food sub-categories (breads and cereals; vegetables; fruits; butter and oils; seafood; meat; milk, cheese and eggs; sugar, coffee, tea and cacao; grains37 ; tubers; food prepared away from home; non-alcoholic beverages and alcoholic beverages) and 6 health sub-groups (consultations, medicines and treatments, tests and analysis, dentistry, ophthalmology and hospitalization and surgery).38 In addition, this survey gathers very useful information on whether or not any household member received a Pensión 65 subsidy in the last three months. Table 3.6 presents some descriptive statistics of this sample, with the first two columns displaying mean and standard error estimates for the full sample of households with adults between 55 and 75 y/o.39 52% are located in rural areas and. They are, on average, composed, of four members, with an important representation, as expected, of children under 5 y/o and adults over 55. Household heads are, on average, 61 y/o, one third are women and they have 3.75 years of education. Their annual consumption is around 5,000 Soles (791 US Dollars) and one fifth have at least one member receiving a Pensión 65 subsidy. Monetary expenses are about 3,000 Soles, of which 68% correspond to food expenses, 4% to health, 3% to education and 2% to housing. Most relevant food sub-categories are: breads and cereals (454 Soles); meat (317 Soles), vegetables (236 Soles); milk, cheese and eggs (205 Soles) and fruits (174 Soles). 3.3.2 Children under 5 y/o Children data is extracted from the 2015 ENDES, a survey sampling woman of childbearing age, collected between March of 2015 and June of 2016. It collects data on several items, including a very complete module about the nutrition and health status of their children under 5 years old. Following earlier studies, children’s nutrition and health is measured by the follow- 33 In Section 3.4 I give the reasons why I selected this bandwidth. Includes tuition, uniforms, books and other supplies. 35 Refers to furniture and household equipment (radios, televisions, computers, blenders, etc.) acquired in the last 12 months. 36 Includes services, rent and imputed rent 37 Mainly legumes like lentils and beans. 38 All these variables are measured in per-capita terms. 39 In order to give some context to these figures and to facilitate comparisons, the Appendix presents similar tables including statistics for the complete sample surveys and shows mean difference estimates with this specific sub-sample. 34 3.3.2 Children under 5 y/o 145 ing indicators: weight for age40 ; height for age41 ; weight for height42 ; body mass index (BMI)43 ; nutritional level44 and anemia levels45 . In addition, as in the 2015 ENAHO, this survey also compiles information on whether or not household members actually benefit from Pensión 65. As in the household level case, here I also restrict the analysis to children in extremely poor households 46 and living with at least one eligible adult between 55 and 75 years old. The final sample is composed of 269 children (Table 3.5). Table 3.7 displays some descriptive statistics of the children integrating the sample (i.e. boys and girls under 5 years old, extremely poor and living in households with at least one eligible member between 55 and 75 years old). 12% live in rural households, which are composed, on average, of 6 members: 1.66 adults above 55 y/o, 1.5 adults between 20 and 54 y/o, 1 children between 6 14 y/o and 1.4 over 5 y/o. Household heads are, on average, aged 62 y/o, one third are women and they hold almost 3 years of education. Children are, on average, 32.7 months old (almost 3 years) and half of them are females. Regarding their nutritional and health status, when it comes to long term measures, like weight for age (Wt/A) or height for age (Ht/A), they are, respectively, 0.49 and 1.36 standard deviations below the median of the reference population. This is not surprising considering that these two indicators reflect past deprivations and illnesses that are usually associated with poor socioeconomic conditions. More worrying, stunting incidence is around 20%, meaning that 2 of every ten children of the sample are, at least, 2 standard deviations below the median height of their age/sex group. Specialized literature warns about the potential effects of these nutritional deficiencies. Stunting, for instance, may lead to increase morbidity and mortality, reduce cognitive and brain development or lead to have short adult stature and later chronic diseases.47 In contrast, the positive mean z-score associated to weight for height (Wt/Ht), 0.41, a measure of short-run 40 Z-score following WHO reference standards (WHO, 2006 ). Z-score following WHO reference standards (WHO, 2006). 42 Z-score following WHO reference standards (WHO, 2006). 43 Z-score following WHO reference standards (WHO, 2006). 44 Probability of: acute malnutrition (weight for age z-score under -2 standard deviations of the median reference standard for their age), stunting (height for age z-score under -2 standard deviations of the median reference standard for their age), underweight (weight for height z-score under -2 standard deviations of the median reference standard for their age) or obesity (weight for height z-score over 2 standard deviations of the median reference standard for their age). These definitions are based on national standards (ENDES, 2014). 45 Probability of : Severe anemia ( level of hemoglobin concentration of less than 7.0 g/dl), moderate anemia (level of 7.0-9.9 g/dl) or mild anemia 46 Extremely poor households were identified following a very similar process to that used by program officials to built the list of potential Pensión 65 beneficiaries (Subsection 3.2.2). Unfortunately, this survey does not have neither information on labor income, nor information on water and electricity consumption. Thus, the identification process of the extremely poor only could followed the IFH index rule. 47 For instance Allen et al. (2001), WHO and UNICEF (2008) and Mongkolchati et al. (2010). 41 146 Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru nutritional status, might be reflecting recent weight gains, probably from the benefits of Pensión 65. Finally, prevalence estimates suggest that, although severe anemia is very rare in the sample, over 30% of the children were at least mildly anemic. Despite the fact that, on this, the WHO reports suggest that most cases of mild anemia do not cause major complications, if it is untreated, it can lead to other health problems, including delayed growth and development and infections. 3.4 Empirical Strategy To estimate the relationship between Pensión 65 and monetary spending and its association with the health of co-resident children, I perform a regression discontinuity analysis, consisting in comparing eligible households and eligible household members under 5 years old, across the eligibility age threshold of the program, i.e. 65 years old. The same technique has been already validated in earlier studies evaluating similar programs, like Duflo (2000, 2003), Martinez (2009), Edmonds et al. (2005), Edmonds (2006), Galiani et al. (2016) and de Oliveira and Kassouf (2012), among others. The main analysis is based on the OLS estimation of a reduced-form given by Equation (3.2): yh/i = ↵h/i + h/i 1 {AOh m >= 65 }+ f ( ÃOh m )+ f ( ÃOh m ) ⇤1{AOh m >= 65} + xh/i + μh/i (3.2) where yh/i represents the outcome of interest for household h or individual i, depending the case. AOhm is the running variable, i.e. 65 years old, measured in months from the 65th birthday of the oldest member of the household. For example, 0 means the oldest member of the household is 65 years old, -120 means she is 55, 120 means she is 75, and so on. 1{AOhm >= 65} identifies cases after the threshold. f (ÃOhm ) is a quadratic polynomial function of AOhm that I also interact with the age indicator (1{AOhm >= 65}), to allow the slope of the age profile to vary on either side of the 65-years-old-cut-off point. xh/i represents a vector of household and individual controls. Finally, μh/i is an error term. In addition, given the non-deterministic character of the age assignment rule associated to the intervention (fuzzy regression discontinuity design FRDD), I also estimate 2SLS regressions of the relationship between the whole set of outcomes yh/i and actual program participation P en65h ; where this variable is a dummy equal to one if at least one household member actually benefits from from Pensión 65. The first stage is given by Equation (3.2), with P en65h being the 3.4 Empirical Strategy 147 dependent variable, and the second stage is given by Equation (3.3). ˆ h + xh/i + μh/i yh/i = ↵h/i + ⇢P en65 (3.3) Before continuing, it is important to clarify that, in this case, the cut-off was decided at December of 2013 (i.e. the running variable is the age of the oldest household eligible member at this date). Other points are off course plausible, given that Pensión 65 was launched in October of 2011 and that the data used in the analysis were collected between January and December of 2015 and March of 2015 and June of 2016. However, as it was mentioned in Subsection 3.2.2, although the program started earlier in some districts, it was only by December of 2013 when it reached universal coverage. In addition, households usually take some time to change consumption patters and the nutrition and health status of children evolves gradually, so take more recent points, like 2015, will not be informative about these behaviors. Anyhow, Subsection 3.5.4 analyzes other possible points, concluding that the December of 2013 threshold is the one that better fits of the data. Following the specialized literature, this equation is estimated using robust standard errors clustered at the level of the running variable (age of the oldest member by December of 2013)48. Household controls include: household composition by 8 age-groups (0 5 y/o, 6 11 y/o, 12 19 y/o, 20 29 y/o, 30-39 y/o, 40 54 y/o, 55 75 y/o and more than 75 y/o); sex, age and education (years of education) of the household head; a dummy equal to 1 if the household is located in a rural area; 20 province dummies and 12 survey round dummies. Individual controls are sex and age (in months). The impact of Pensión 65 on yh/i is given by h/i , which represents the difference in the intention- to-treat probability on either side of the threshold. The crucial assumption is that in the vicinity of the 65-years-old cut-off, households and children differ only in their probability of receiving the program, so observations just missing the age threshold are a good counterfactual for their age eligible counterparts (i.e. local continuity assumption). This assumption could be violated if households and children around the age eligibility threshold have different characteristics, that are also correlated with outcome variables. For instance, having a member over 65 y/o could be a sign of healthier household members with better nutrition habits and health practices. In this case, estimation coefficients from Equation (3.2) might be confounded with these positive attitudes towards health, instead of being the pure impact of the program. 48 See for instance Lee and Card (2008). Tables 3.6 and 3.7 compare age-eligible and non-age eligible households and children under 5 y/o. The first table displays some very small, but still significant, differences, in terms of household composition, specially among the groups between 6 and 54 years old. Eligible household heads are also almost 7 years older and have, on average, 0.8 more years of education. Consistently with the characteristics of the program and its expansion time-table, half of them have received a Pensión 65 subsidy in the last 12 months, whereas this percentage is only 1% among households in which the oldest eligible member was less than 65 y/o by December of 2013. Finally, eligible households spend more in breads and cereals, vegetables, education services, two health categories (tests and analysis and dentistry) and durables. The second table shows that children in households with an eligible member have very similar characteristics than those in households with younger members. They are, however, slightly different in terms of the household composition, more precisely in the number of adults between 30 39 y/o and in the number of elderly over 75 y/o, and in the education of household heads. The local continuity assumption will also be violated if individuals can control the assignment variable, i.e. living with persons holding 65 years old by December of 2013. Manipulation might be an important concern in this case because, due to the program, households may have incentives to change their composition by, for instance, moving with potential eligible elderly relatives or friends, just to receive the subsidy. In order to test the validity of the continuity assumption I use a McCrary Test (McCrary, 2007). Figure 3.4 shows the results. The test fail to reject the null hypothesis of no discontinuity at the threshold, for both samples, suggesting that there is no systematic change in the density of eligible individuals on either side of the discontinuity. Although in the household sub-sample (Figure 3.4 Graph A) there appears to be a slightly lower density to the right of the threshold, this difference is not significant and goes in the opposite direction of the manipulation hypothesis. Another very sensitive issue in this setting is the choice of the window of observations at both sides of the cut-off point. Al thought, in principle, the closer to the cut-off, the better, a very narrow window, risks of leaving very small samples reducing the power of the estimations. Imbens and Kalyanaraman (2012) suggest a bandwidth choice test to perform for each sub-sample and outcome of interest. In order to have some homogeneity along the different regressions, all the estimations are run taking the average of the different optimal bandwidths suggested by this procedure: 119 months, which corresponds to the rank [55 75] years old. Other concerns related to the sensitivity of the specifications and the validity of the strategy will be resumed at the end of Section 3.5. 3.5 Results This section presents and discusses the main findings drawn from the empirical analysis outlined before. First, I report the estimates for the association between Pensión 65 and household monetary spending and then, I document the its relationship with the nutritional and health status of young children. Beforehand, I provide evidence of the correlation between age eligibility and program participation. Figure 3.5 plots, in two separate graphs, the association between actual program participation (prob ability of being treated ) and the age of the oldest household member by December of 2013 (in months from her 65th birthday), for the two samples of interest: households with children under 5 years old and children under 5 years old. Dots represent the fraction of households and children under 5 y/o actually receiving a Pensión 65 subsidy , and the blue curves represent the correspond ing quadratic prediction plots.49 on either side of the age cutoff. Households whose eligible members were 65 years old, or more, by December of 2013 are represented on the right side of the da shed red line. Both graphs suggest an upwards jump in the probability of being treated around zero (65 years old), evidencing a discontinuity when the eldest household member becomes eligible to participate in Pensión 65. Formal estimations of Equation (3.2) for Pensión 65 participation (Table 3.8) show that the probability of treatment is 22 percentage points higher among households with age eligible members, in the sample of households with children under 5, and 41 pp, in the sample of children under 5 years old. 3.5.1 Household Monetary Spending Figure 3.6 illustrates graphically the relationship between total monetary spending, including 25 different sub-categories, and the age of the oldest household member (in months from her 65th birthday) by December of 2013.50 Several graphs show an upwards jump at the threshold when barely age eligible and non-eligible households are compared, suggesting a positive relationship between Pensión 65 age-eligibility and total spending (Graph A) and two of its main components, food (Graph B) and health spending (Graph R). 49 Calculated from a linear regression of yh on AOhm and A2Ohm. Dots indicate mean values and the blue curves represents the corresponding quadratic prediction plots, calculated from a linear regression of yh on AOhm and A2Ohm , on either side of the age cut-off. Intra-household Income Transfers and its Effects on Children’s Nutrition and Health in Peru This positive discontinuity is also the trend observed for the main food spending sub-groups, for instance, breads and cereals (Graph C), vegetables (Graph D) and grains-legumes (Graph L); and the most relevant health spending categories, health consultations (Graph S) and medicines and treatments (Graph T). In order to confirm these results, Table 3.9 provides formal regression results for Equation (3.2). Each line corresponds to a different estimation, the dependent variable appearing in rows. The first three columns display OLS h coefficients, standard er- rors and the corresponding R2 . Columns 4 and 5 report the results (Chi-squared and P-value) of a multiple hypothesis test (using the Bonferroni correction) implemented in order to check the statistical relative importance of each of the outcomes. The estimates corroborate the positive correlation between Pensión 65 and spending in vegetables, grains-legumes and health services. OLS coefficients indicate that households with ageeligible members spend, on average, 1,123 more Soles/year compared to similar households with eligible members below the age threshold. Relative to the amount of the program allocation, this spending increase is equivalent to 75% of the annual maximum Pensión 65 allocation that an individual can get. In the specific case of total food spending, although the coefficient is positive and large, it is not statistically significant at the 10% level. However two sub-categories show positive estimates: vegetables (117 Soles, 8% of the maximum program allocation) and grains-legumes (30 Soles, 2% of the maximum allocation). The positive correlation of the Pensión 65 subsidy with these specific categories of food spending is important for two main reasons. The first is that both vegetables and grains-legumes, are highly recognized for their nutritional relevance in the diet of young children51 , given their high fiber, vitamin A and vitamin C contents.52 . The second is that, these food categories are often less accessible for low income households (Stewart et al., 2003). Given that households of the sample are disproportionately poor and have lower consumption and food spending, compared to national standards (see Table A3.1 of the Appendix), the marginal utility of increasing these calories could be highly significant for them, bringing about more powerful nutrition and health benefits. When it comes to access to health services, estimates indicate that age eligible households surpass the total spending of non-eligible households by 123 Soles (8% of the maximum allocation), explained mainly by an upsurge in consultation expenses (13 Soles, 0.9% of the allocation). In Peru, limited access to health care is one of the main causes of child mortality, esti51 52 Specially those between 6 and 59 months old, in complement to breastfeeding. See for instance: WHO and UNICEF (2003) and WHO and UNICEF (2008) 3.5.1 Household Monetary Spending 151 mated at 13.5 deaths per thousand live births in 201553 . In addition, despite significant efforts in terms of coverage, household income is still the main source of financing health services (PAHO, 2012). These elements make the poorest most vulnerable to health shocks, explaining the positive impact of income transfers, like Pensión 65, in improving access to health care. These results are consistent with previous findings related to similar cash transfer programs targeted to the elderly, including the ones on Galiani and Gertler (2016) on Pensión 6554 and Martinez (2009) about the program BONOSOL in rural Bolivia55 They also contribute to strengthening the hypothesis that income constraints play a key role in determining the quantity and the quality of food expenditure and the access to health care services in developing countries, pointed out already by impact assessments of other cash transfer programs. For instance, also for Peru, Vakis and Perova (2009) show that the program Juntos has positive effects on several food categories and in the intensity of use of different health services. In order to verify whether or not these findings are specific to the sample of households with children under 5 years old, and to compare them with other household spending strategies, I reestimate Equation (3.2) for the same set of monetary spending categories, across the following three sub-samples : (i) households not reporting any children under 5 y/o, (ii) households reporting at least one woman between 15 and 49 y/o and (iii) households reporting at least one adult over 65 y/o (in 2015).	32,182
https://openalex.org/W3188087457_1	Spanish-Science-Pile	Open Science	Various open science	2,021	Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México	None	Spanish	Spoken	7,716	13,634	Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México1 Tradition against hegemonic development, axes of a political conflict: The case of the Huexca mega-project in Mexico Alfonso Barquin Cendejas Instituto Nacional de Antropología e Historia, México albarcen@gmail.com Mariana García Galicia Escuela Nacional de Antropología e Historia, México garciamariana23.94@gmail.com Resumen El presente estudio, basado en trabajo etnográfico y documental, expone un panorama general del conflicto político surgido en Huexca, Morelos, México, a propósito de la construcción en la comunidad de una planta termoeléctrica que alcanza más de una década de confrontación. Se analizarán dos cuestiones fundamentales, por un lado, el accidentado proceso de su implantación de manera impositiva, y por otro, las consecuencias de una autoritaria actuación con la población. No consultar a la población local generó dinámicas internas que dividieron a la población entre los que simpatizaban con el proyecto y los que no, lo que ha derivado, a la fecha, en la imposibilidad del funcionamiento de la planta. Esta división es un subproducto perverso de la implantación de grandes proyectos de infraestructura por la falta de una consulta eficaz que reafirme y unifique a las poblaciones en su aceptación o rechazo. Palabras clave: Megaproyecto, desarrollo hegemónico, conflicto político, división local. Summary This paper, based on ethnographic and documentary work, presents an overview of the political conflict that has arisen in Huexca, Morelos, Mexico, regarding the construction of a thermoelectric plant in the community, which has been going on for more than a decade of confrontation. Two fundamental issues will be analyzed: on the one hand, the difficult process of its imposing implementation and, on the other hand, the consequences of an authoritarian approach to the population. The failure to consult the local population generated internal dynamics that divided the population between those who sympathized with the project and those who did not, which has resulted in the impossibility of the plant’s operation to date. This division is a perverse by-product of the implementation of large infrastructure projects, due to the lack of effective consultation to reaffirm and unify the populations in their acceptance or rejection. Keywords: Megaproject, hegemonic development, political conflict, local division. Artículo: Recibido el 20 de agosto de 2020 y aprobado el 15 de febrero de 2021 Cómo citar este artículo: Barquín Cendejas, A. & García Galicia, M. (2021).Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México. Reflexión política 23(47), pp. 42-58. doi: https://doi.org/10.29375/01240781.4083 1. Se agradece a la comunidad de Huexca la información para la redacción de este artículo. 42 REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) Introducción En la construcción de obras que requieren las poblaciones humanas como consecuencia de sus necesidades, las grandes obras o megaproyectos se presentan como objetos problemáticos. Todo tipo de circunstancias de alcance regional o nacional presionan para su implantación sobre un territorio y, por otro lado, su aterrizaje en un enclave limitado impactan de manera dramática a las poblaciones del lugar. Consideraciones de tipo económico, político, jurídico, cultural y ecológico giran en torno de su ocurrencia, pues los beneficios económicos generales y en la zona, se contraponen con los mecanismos políticos por los que se consiguió implantar el megaproyecto, la legalidad o no de dicho proceso, las confrontaciones entre visiones culturales sobre dicha obra y el daño o mitigación de los efectos en el medio ambiente. Estas características perfilan el problema de esta investigación: ¿cómo abordar los múltiples conflictos derivados del desarrollo de obras de tal magnitud que suponen intereses económicos y políticos de gran alcance frente a necesidades locales no precisamente afines a tales megaproyectos? Frente a este dilema se requiere algún tipo de mediación política y, por tanto, se pueden desplegar mecanismos de diálogo, consulta, presión o cooptación que es necesario comprender Estas consideraciones que surgen a propósito del desarrollo de un megaproyecto en la región del centro de México y su culminación en la comunidad de Huexca dan pie al abordaje de dos preguntas de investigación. En primer lugar, se busca dilucidar cómo es el accidentado proceso que va desde la concepción hasta la finalización de las obras y por qué no se ha podido iniciar su operación. En segundo lugar, ¿por qué los nulos o deficientes mecanismos de consulta a la población afectada han determinado, no sólo la imposibilidad de iniciar la operación de la termoeléctrica, sino también la ruptura de la armonía comunitaria? A partir de información documental, trabajo de campo y entrevistas con actores relevantes entre 2017 y 2019, se darán luces sobre estos temas, con el fin de buscar evidencia de las fallas más evidentes y las consecuencias producidas. 1. Marco teórico y antecedentes La idea de “megaproyecto” no es en sí una idea nueva o novedosa. Cualquier sociedad con dimensión estatal a lo largo de la historia se ha visto obligada a emprender esfuerzos excepcionales y fuera de lo común que afectan el medio para resolver las problemáticas sociales que se derivan de la concentración de población y la diversificación de actividades humanas. Tan patente es este hecho que ya desde el Anti-Dühring, Engels (1968) elabora un modelo para comprender la complejización social derivado de las percepciones de Marx (1976) del “modo de producción asiático” en el que se perfila la separación de las clases entre los trabajadores generales y aquellos que se encargan de la dirección de las grandes obras hidráulicas necesarias para mantener la viabilidad del grupo social. Caso semejante al de Karl Wittfogel (1966), que dedica toda una obra al estudio pormenorizado del despotismo oriental, en el que las grandes obras hidráulicas constituyen uno de los núcleos definitorios de la manera en la que se organizan esas sociedades, tanto social como políticamente. Destaca también la visión contemporánea de Linz Ribeiro (1987, 2006, 2007) y su propuesta de análisis de los “proyectos de gran escala”, sus implicaciones sobre el desarrollo y las relaciones de poder que involucran. Estas consideraciones iniciales vienen a lugar porque se insiste que la idea de megaproyecto dista de ser nueva en la historia de las sociedades estatales y su efecto es de tal naturaleza que alcanza dimensiones civilizatorias. Entonces, surge la pregunta del por qué en los años recientes se han incrementado tanto el desarrollo y la necesidad de estas grandes obras, así como las tensiones que surgen en las comunidades aledañas a estos megaproyectos. Es evidente que las presiones de la globalización contemporánea empujan a los actores económicos a la búsqueda incesante de una tasa de ganancia, más aún en el capitalismo actual de tinte neoliberal en el cual el control social de los métodos de obtención de ganancias busca, aún más, subordinar a los poderes políticos a las reglas del libre mercado; lo que Acselrad (2006) define REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) 43 Alfonso Barquin Cendejas, Mariana García Galicia Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México como una reconfiguración de los poderes políticos a partir de la globalización y la reestructuración productiva. Estas reconfiguraciones, los conflictos que se producen en las sociedades y el daño en el medio ambiente, han dado lugar a lo que Martínez Allier (2015) define como el objetivo de la ecología política: “dar cuenta de cómo el poder político incide en los conflictos socio ambientales” (p. 57). Tales conflictos contemporáneos inciden no solo en los ámbitos económico-políticos y ecológicos sino también, en los culturales. Esta es una de las líneas de reflexión que abre Escobar (2006) sobre las luchas de las comunidades frente a las presiones productivas de la globalización, porque enfrenta las visiones locales o tradicionales con los procesos productivos globalizados. Con el fin de entender qué es lo que ha hecho que en el presente los megaproyectos sean tan dañinos y generadores de conflicto, además de lo expuesto arriba, se pueden encontrar otros argumentos que hacen su implementación aún más conflictiva que en épocas anteriores. Uno de los principales es el cambio dramático de los patrones de crecimiento de la población mundial que inició con una cuesta de crecimiento muy empinada hacia los años cincuenta del siglo XX, derivada de varios factores, entre los más importantes se encuentran: el inicio de un ciclo de paz luego de las dos guerras mundiales; la estabilidad social que produjo en todo el mundo un gran periodo de auge económico por casi treinta años y que Hobsbawm (1995) llamó “la edad de oro”; la expansión y el mejoramiento dramático de los sistemas de salud estatales, en particular la síntesis y disponibilidad en gran escala de antibióticos y vacunas; el incremento general de los niveles educativos y, por lo tanto, la mayor especialización y productividad de los individuos; el proceso acelerado de mecanización y robotización de la producción industrial y la incorporación de la inteligencia artificial tanto en lo productivo como en lo administrativo. Estos cambios potenciaron una dinámica cultural de gran consumo, centrado no solo en las necesidades esenciales de los humanos, sino también en la satisfacción por el consumo mismo de las mercancías, lo que generó en el ámbito industrial una búsqueda desmedida por la ganancia. Finalmente, que, por primera vez en la historia de la humanidad, más gente vive en las ciudades que en el campo, lo que estimula un modelo basado en el consumismo, pues la gran cantidad de personas concentradas en un punto, incrementa las necesidades de mercancías de todo tipo y, por tanto, la infraestructura para satisfacer tal demanda se presenta como colosal. Basta decir que hacia 1960 la población rondaba los 3,000 millones de personas, y que para 2050 se habrá multiplicado por tres. Este crecimiento, además, está transformando dramáticamente al mundo, pues la proporción rural-urbana pasará, según la Organización de las Naciones Unidas (ONU), de dos a uno en 1950, a invertirse cien años después; es decir, 6,000 millones en las ciudades y 3,000 en el campo para el año 2050 (ONU, 2018). Por otro lado, a partir de los años 90 del siglo XX, el deseo de consultas que exponían y legitimaban la posición de las poblaciones frente a estos proyectos, se ha incrementado dramáticamente en América Latina; reformas jurídicas para establecer mecanismos de democracia directa y consultas efectivas han ocurrido de manera importante en toda la región (Altman, 2005; Lissidini, Welp & Zovatto, 2008). Esto supone una nueva realidad para la ocurrencia de los megaproyectos. Por un lado, las poblaciones son más conscientes de la existencia de mecanismos que las protegen frente a los megaproyectos y que determinan la necesidad de informarles, previa y verazmente, las ventajas y desventajas de que se les consulte libre y públicamente y que sea respetada su opinión. Por otro lado, esta nueva realidad ha hecho conscientes a los actores de la necesidad de gestionar adecuadamente la realización de los proyectos (López & Shane, 2014; Alberti & Pereyra, 2018). No obstante, este nuevo esquema 2. La novedad de estos problemas y, por lo tanto, su análisis, llevó a Enrique Leff (2006) a aseverar que uno de los campos que los abordaban, la ecología política, estaba apenas en construcción. 44 REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) no garantiza la ideal utilización de los instrumentos de consulta, pues como muchos otros mecanismos, los partidos políticos o los propios gobiernos han invadido el territorio gracias a las posibilidades de intermediación, manipulación o justificación al margen de los intereses de los afectados (Altman, 2010; Welp, 2010; Welp & Serdült, 2014). Frente a este contexto que perfila lo complejo de los megaproyectos en el contexto de las relaciones hombre-naturaleza y, al mismo tiempo, el surgimiento de líneas que exigen mecanismos de consulta popular y participación informada, es que se pueden exponer ciertos antecedentes sobre este tipo de obras en América Latina y en particular en México. Un caso emblemático es el de las hidroeléctricas que durante mucho tiempo – remontándose incluso a la época colonial- han afectado a las poblaciones rurales, en particular a las poblaciones indígenas de América Latina (Bartolomé, 1992; Gómez et al., 2014). Destaca aquí que siendo la mayoría de los megaproyectos ubicaciones en territorios campestres o no urbanos, los pueblos indígenas son los más susceptibles de verse afectados. Justo la relación entre un entorno ecológico, su aprovechamiento masivo y pobladores afectados incide de manera problemática a los indígenas, como fue exhibido de manera relevante por Boege (2008). El resultado final de este tipo de obras ha tenido en México resultados dispares: un ejemplo es el de las grandes afectaciones por la presa Cerro de Oro 1974-1988 (Bartolomé & Barabas, 1990); otro es el de la presa La Parota, iniciada en 2001, que por las agresiones estatales y los conflictos no resueltos (Chávez, 2009) a la fecha sigue sin terminarse. En todo caso, como se reseñó anteriormente, la historia de la cultura de consulta y negociación es muy joven en toda América Latina. Para el caso mexicano, la reforma en materia de derechos indígenas a la Constitución Mexicana en el año 2001, derivada de las presiones por ajustarse al espíritu del Convenio 169 de la Organización del Trabajo, estableció la necesidad de operar las obras que los afectan en coordinación con ellos pero sin un mandato que obligara consultas vinculantes.3 A partir de esto se aceleró el movimiento para hacer de la consulta un mecanismo regular, aunque con resultados todavía decepcionantes, pues el escaso aparato legal se cruza con la intensa demanda ciudadana (Zaremberg et al., 2018). Así, casos como el del acueducto “Independencia” que en 2010 buscó apropiarse de un caudal de agua que surte al pueblo Yaqui sin mediar una consulta y que, por sentencia de la Suprema Corte de Justicia, se mandató una consulta aun no realizada (Gómez, 2017; Lerma, 2014). En ese sentido, el caso de Huexca pertenece a esos megaproyectos no consultados y no concluidos. Últimamente se han realizado algunas consultas en el país, derivadas de una nueva visión de gobierno del presidente López Obrador. Sin embargo, todas estas consultas que han tenido su origen en intereses de gestión del gobierno actual, han sido duramente criticadas por desaliñadas, incluso por las Naciones Unidas, y como se señaló anteriormente, han tenido más un papel de justificación de intereses gubernamentales que de conocer la opinión genuina de los afectados. Ese es el contexto en el que se realizó la consulta del megaproyecto en Huexca y como se verá; resultó en un ejercicio inútil y que mantiene el conflicto en marcha. Así pues, el desarrollo de los megaproyectos tiene lugar entre intereses civilizatorios, económicos, políticos, locales y culturales que se entreveran de manera conflictiva, pues en cada uno de esos niveles se promueven o desalientan los megaproyectos. El caso de Huexca expone dos momentos de este proceso. Al inicio, se buscó por todos los medios y se logró evitar cualquier mecanismo de consulta que impidiera o modificara el programa de toda la obra, limitándose en su primera época a informaciones vagas y beneficios aislados en la población. Al final, con la obra terminada, pero en paro, el gobierno actual realizó una polémica consulta diseñada para justificar la operación de todo el sistema, que no fue reconocida por amplios sectores de la región, que ocurrió días después del asesinato del líder de los opositores a la megaobra y que fue cuestionada por el órgano nacional de vigilancia de los derechos humanos. Por tanto, el mecanismo de consulta falló al inicio por no ocurrir, y al final por implementarse de manera amañada. Las consecuencias de todo lo anterior es una obra parada a la fecha y una comunidad dividida. 3. Constitución Mexicana. Disponible en: http://www.diputados.gob.mx/LeyesBiblio/pdf_mov/Constitucion_Politica.pdf 45 Alfonso Barquin Cendejas, Mariana García Galicia Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México 2.- Los ángulos del conflicto Esta sucinta introducción busca dar sentido a las tensiones que genera la implantación del Proyecto Integral Morelos, megaproyecto regional de energía que supuso la construcción de una planta de generación eléctrica en la pequeña localidad de Huexca en el estado de Morelos, México, y cuya población es de 1,153 habitantes (INEGI, 2021). La obra en cuestión consiste en una central termoeléctrica de ciclo combinado, que supone 2 turbinas de gas y dos generadores de vapor para la producción de electricidad. La instalación que está asentada en un predio de la comunidad, toma su carácter de “megaproyecto” en el sentido de que no es solo una afectación de carácter local, sino que impacta toda una región del estado de Morelos y, por lo tanto, los efectos dañinos también se extienden a decenas o cientos de kilómetros de la obra final. Por ello, todos los alcances y resistencias al proyecto están englobados en lo que se llama “Proyecto Integral Morelos” –en adelante PIM-, que buscó no sólo la construcción y operación de dicha termoeléctrica que produciría alrededor de 620 mega watts, sino todo un desarrollo regional al introducir a través de un gasoducto energía suficiente para el desarrollo de una amplia zona industrial del Estado de Morelos. Así, según la Manifestación de Impacto Ambiental (MIA) elaborada por la Comisión Federal de Electricidad (CFE), el Proyecto consiste en: • Una central termoeléctrica combinada en la localidad de Huexca. • Un ducto de gas de 48’ proveniente del estado de Tlaxcala. • Un acueducto hacia la termoeléctrica con el fin de servir para la generación de vapor y el enfriamiento del sistema. • Una red de transmisión eléctrica hacia la subestación de Yautepec, Morelos (CFE, 2011a). Como se puede ver, la obra compromete muchos esfuerzos y, al mismo tiempo, muchos impactos en las zonas que atraviesa. Es decir, las afectaciones al medio y a las poblaciones aledañas a todo el proyecto deberían ser evaluadas mediante información suficiente para cuantificar los daños y los beneficios que se desprenderían de la obra. Sin embargo, no se realizó consulta alguna a la población afectada, únicamente se efectuaron asambleas informativas y se repartieron algunos apoyos a los pobladores cuando iniciaron los trabajos en el año de 2012. Este pecado inicial, el de no consultar el arranque de la obra, fue acumulando agravios hasta que se generó gran descontento a lo largo de todo el recorrido del PIM. En la trayectoria del gasoducto hay conflictos por la cercanía con zonas habitadas; en el acueducto también, debido a que no se quiere respetar la restricción al uso exclusivo de aguas negras, lo que ha motivado acciones jurídicas para tratar de detener las obras; y en Huexca, a la fecha, tampoco se ha permitido iniciar la operación de la planta. Este rápido panorama busca resaltar cómo es que todo el proceso derivó en un conflicto interno en Huexca, pues la población se dividió entre aquellos que apoyaban la apertura de la planta – denominados por sus rivales como “los termos”- y aquellos que rechazaban la obra- denominados los “antitermos”. Todas las instancias de coordinación local: sistema de cargos para la fiesta, asambleas vecinales, comités escolares y órganos de representación política local, se vieron inundados por este antagonismo, dañando la tranquilidad y armonía relativa que existía en Huexca. Diversos factores que analizaremos, llevaron a tal punto las cosas, que la polarización culminó en situaciones de tensión y violencia fatal. El propósito de este trabajo es mostrar las consecuencias originadas por la ausencia de consulta a la población, o peor aún, por una consulta deficiente y en condiciones políticas impropias, lo que ocasionó la falta de consenso sobre los beneficios y los daños de la obra. De esta manera, ni los bandos que se oponían pudieron encontrar razones para aceptar la obra, ni los simpatizantes, razones para mitigar su efecto, por lo tanto, todo el PIM es visto entre muchos sectores como ilegítimo. Adicionalmente, contiene un elemento de emotividad política. En el 2014, el entonces candidato a la presidencia de México, Andrés Manuel López Obrador, visitó el estado de Morelos y aseguró a los habitantes que si él ganaba la elección no pondría en operación la termoeléctrica. Habiendo triunfado en las elecciones regresó ya como presidente en funciones a principios de 2019 y declaró que la no operación de la termoeléctrica era un desperdicio de dinero y que habría que aprovechar la obra. El cambio de criterio enfureció a los que creyeron en su palabra y polarizó aún más la situación. López Obrador propuso la realización 46 REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) de una consulta, el presidente acataría el resultado. No obstante, la consulta fue diseñada de manera que el peso cuantitativo de los afectados locales fuera mínimo, al incluir municipios de Tlaxcala, Puebla y Morelos, cuyas tazas de afectación no eran semejantes; incluso se incluyeron ciudades como Cuernavaca o Cuautla, en donde las afectaciones son casi inexistentes y los beneficios máximos, pues se prometió bajar el precio de la luz cuando funcionara la obra. La consulta arrojó un resultado aproximado de 59% a favor y 41% en contra, como se aprecia, no es un resultado muy contundente y, adicionalmente, como se dijo, de diseño sesgado. Hay un elemento adicional, unos días antes de la consulta fue asesinado Samir Flores, el líder principal de la oposición a todo el PIM, en circunstancias y motivos que a la fecha no son conocidos. Esto alimentó la sospecha y desconfianza hacia la consulta, pues sin haber condiciones políticas y de seguridad para tal ejercicio, el ejecutivo federal decidió instrumentarla. De este modo, surgió de nuevo el rechazo y la indignación por una consulta que favorecía los intereses del gobierno, lo que tiene a la termoeléctrica detenida y a todo el PIM en suspensión. En adelante se darán detalles puntuales del PIM y de los efectos concretos en la localidad de Huexca. 3.- La imposición en Huexca En los antecedentes del caso figuran los inicios del diseño y la planeación del Proyecto Integral Morelos. Hacia el año 2009 el entonces gobernador de Morelos, Marco Adame, presentó ante la Comisión Reguladora de Energía una solicitud para contar con infraestructura de suministro de gas natural hacia su estado, para el beneficio y expansión de las zonas industriales. Emitida la resolución aprobatoria, en 2010 la Secretaria de Energía federal (SENER) instruyó a la Comisión Federal de Electricidad (CFE) para que a la par de esos trabajos se instalara una central de generación eléctrica alimentada por gas natural. Del mismo modo, en noviembre de ese año el gobierno estatal a través del Sistema Operador de Agua Potable y Saneamiento de Cuautla (SOAPSC) emitió una autorización para poner a disposición de la planta hasta 280 litros de aguas negras por segundo para su utilización en la termoeléctrica; con estas disposiciones se pudo elaborar el contrato que 47 la CFE signó con Abengoa/Elencor. De inicio, no era obligatoria legalmente una consulta para la obra, aunque hay que señalar que a partir de 2014 está establecido en la ley que, para obras de naturaleza energética, se debe contar con la aprobación de las comunidades indígenas afectadas. Dado que ese criterio normativo es posterior, el proyecto arrancó sin la opinión de los involucrados. Según la MIA, se plantearon dos zonas probables en Yautepec y Yecapixtla; sin embargo, por las condiciones de alta productividad agrícola dependiente del riego en Yautepec, se preveía un rechazo a la venta de terrenos, por lo que, considerando la existencia del corredor industrial Burlington-Saint Gobain, se eligió Yecapixtla. Ahí existían tres posibles localizaciones: Cuautlixco, Huexca y Los Laureles, donde se suponía mayor disposición para la adquisición de los terrenos. De acuerdo con el mismo documento, se realizaron todo tipo de evaluaciones en varios índices. Destacan los rubros de abastecimiento de agua, infraestructura eléctrica, pendiente del terreno, clima, población, etc. En estos índices, las tres localidades lucían más o menos semejantes; la gran diferencia se encontraba en los terrenos de ubicación de la termoeléctrica, pues en Cuautlixco y Los Laureles, la propiedad de los terrenos probables era ejidal, a diferencia de Huexca que era privada, donde adicionalmente, se contaba con uso de suelo para infraestructura. Además, en Huexca el terreno estaba a mayor distancia del núcleo urbano y la densidad poblacional era menor. Con todos estos datos, la CFE decidió la implantación de la termoeléctrica en Huexca. Al ser el terreno privado, se evitaba negociar con distintos actores de la comunidad, lo que definió la ausencia de consultas de cualquier tipo. Con estas condiciones, se planeaba terminar la central hacia 2013 y que la operación iniciara hacia 2015. El tema del agua era de especial preocupación. Este renglón impactaba no solo a Huexca sino a toda la cuenca de la región. Un acueducto de 12 kilómetros conduciría 216 litros por segundo, de los cuales, 200 se emplearían para el enfriamiento de la planta y 16 para repuesto del ciclo. En acuerdo con los pobladores de la zona, este caudal estaría constituido únicamente por aguas negras tratadas. A la fecha presente (mediados de 2020), faltan construir 140 metros del acueducto en la zona de la población de Apatlaco a causa de amparos contra la obra, pues en el proceso de Alfonso Barquin Cendejas, Mariana García Galicia Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México finalización, se comenzó a hablar de la necesidad de un mayor caudal, el cual estaría constituido no solamente por agua tratada, sino por agua limpia. Con relación al gasoducto, el cual también requirió una MIA, constaría de un ducto de 76 centímetros de diámetro con una longitud de 160 kilómetros, iniciando en el estado de Tlaxcala, atravesando por Puebla y llegando por Morelos hasta Huexca (CFE, 2011b). El derecho de vía es una franja de 25 metros y la zona de influencia de 500 metros. El flujo diario estimado sería de 337 millones de pies cúbicos diarios de gas natural. Para lo anterior, se procedió a establecer contratos de derechos de paso del ducto en las tierras elegidas, arrojando que se atravesarían en el recorrido final 32 municipios. Actualmente, el ducto está completamente terminado, pero sin funcionamiento, pues su objetivo, suministrar gas a la planta de Huexca, no es necesario por causa de su inactividad. De la línea de transmisión, esta posee una capacidad de 40 kilovatios y una longitud de 20 kilómetros entre la termoeléctrica en Huexca y la subestación de Yautepec. Según la manifestación de impacto ambiental de la línea de transmisión, constaría de 65 torres ubicadas a cada 350 metros de distancia. La obra está concluida y en suspensión desde 2015. De este modo, este megaproyecto consta de 4 grandes rubros: la termoeléctrica, el acueducto, el ducto de gas y la línea de transmisión, que a la fecha están sin funcionar en atención a las fuertes protestas y rechazo de la población por el desdén con el que se trató a la población de las zonas afectadas, las cuales reaccionaron con manifestaciones, plantones y acciones legales que impiden al presente, el inicio de las operaciones. 4.- “La traición” Un tema que es pertinente desarrollar es que la oposición a este megaproyecto no ocurre en el vacío, es decir, está sustentado en colectivos populares que se organizan para defenderse de los distintos temas que les afectan. En el estado de Morelos había, con relación a este tema, un antecedente relevante: en el año de 2003 se tenía la intención de pasar un gasoducto a través del territorio del estado. Las comunidades que se opusieron, se organizaron en el “Frente de Pueblos en Defensa de la Tierra y el Agua de Morelos” (FPDT) impidiendo la instalación.4 Nueve años después, este mismo colectivo constituye el núcleo opositor al PIM. Desde el inicio de las obras, se comenzó a sospechar en virtud de que no había información y todo se basaba en rumores, así que en asociación con la “Asamblea Permanente de Pueblos de Morelos”, los pobladores se fueron organizando para tener información, tanto de lo que sucedía en Morelos, como lo que ocurría en Tlaxcala y en Puebla. Sus tácticas de lucha, que iniciaron en 2012, se conformaban de plantones, marchas, mítines y asambleas informativas, respecto a los alcances y posibles afectaciones de un megaproyecto que desconocían casi en su totalidad. Su actividad opositora les ha costado hostigamiento, detenciones y demandas judiciales por afectación. Incluso, a decir de los activistas, la ejecución del principal líder del movimiento, Samir Flores, que ocurrió días antes de la consulta que finalmente realizó el gobierno federal, cuyo crimen no ha sido esclarecido. Esta última circunstancia ha sido el detonante final de un cambio total dentro del movimiento. En las etapas previas, los activistas luchaban en contra de gobiernos que ideológicamente consideraban antagónicos. Tanto en la presidencia de Felipe Calderón, como en la de Enrique Peña Nieto, el movimiento suponía a estos gobernantes parte de la derecha o del neoliberalismo, y por ende la lucha tenía un orden “natural” dentro de la cosmogonía política contemporánea: Izquierda, Justicia social y Pueblo; frente a Derecha, Neoliberalismo y Elites. Tal orden político se vio reafirmado por el hecho ya mencionado, de que en un mitin de promoción, el candidato natural de la izquierda a la presidencia del país, Andrés Manuel López Obrador, declaró en Yecapixtla, Morelos en 2014, que: “…no queremos ese gasoducto, esa termoeléctrica, y no queremos tampoco las minas que nada más van a destruir el territorio y van a contaminar las aguas” (El sol de Cuernavaca, 2019). Con esa promesa, los miembros de las organizaciones confiaron en el candidato que resultó ganador en las elecciones presidenciales del 2018. Sin embargo, ya en el primer mes de su gobierno, declaró públicamente que el inicio de las operaciones de la termoeléctrica de Huexca no era 4. A consecuencia de PIM que afectaba territorios distintos de Morelos, se integraron a este frente representantes del estado de Puebla y el de Tlaxcala, por lo que pasó su denominación a “FDPT-MPT” incluyendo las siglas de los tres estados. 48 REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) una decisión tomada, que se iba a revisar y que se implementaría una consulta el 23 y 24 de febrero de 2019 para determinar el destino final. Ante esto, los colectivos organizados reactivaron su actividad pública, pero con un cambio. Iniciaron a considerar que el gobierno en turno, más allá de su etiqueta ideológica de izquierda, no estaba de su lado, es decir, los estaba traicionando. En un evento en la ciudad de Cuautla, Morelos, el 10 de febrero de 2019, a dos semanas de la consulta, López Obrador escuchó protestas por la posibilidad de consulta sobre la termoeléctrica y respondió a los inconformes: “Escuchen, radicales de izquierda, que para mí no son más que conservadores, escuchen, si no se utiliza la termoeléctrica de la CFE, de una empresa de la nación, en vez de tener la luz para alumbrar todo Morelos, tendríamos que seguirle comprando la luz a las empresas extranjeras” (Político MX, 2019). Con esto declaraba su distancia ideológica con los inconformes y reafirmaba que el criterio para la termoeléctrica no era el que les enunció en 2014 relativo a la afectación al entorno, sino un criterio de desarrollo industrial nacional por encima de las afectaciones locales. En este ambiente de tensión, en la madrugada del miércoles 20 de febrero, tres días antes de la consulta, Samir Flores fue asesinado de cuatro balazos. López Obrador declaró ese mismo día que lamentaba los hechos, pero que no iba a suspender la consulta sobre la termoeléctrica: “…a lo mejor entre las posibilidades [del asesinato] era afectar la realización de la consulta” (BBC News Mundo, 2019). Como se mencionó anteriormente, el asesinato de Samir polarizó totalmente el ánimo en contra del gobierno federal y definió el sentimiento de traición por parte del presidente de la República. Era evidente en la consulta, tanto el formato proclive a las intenciones del gobierno como la falta de condiciones políticas para su celebración. El mismo día de los hechos, la Comisión Nacional de Derechos Humanos (CNDH) por voz de su presidente, declaró que no había condiciones aceptables para una consulta de acuerdo con los estándares nacionales e internacionales. Se realizaron dos marchas el viernes 22, un día antes de la consulta para protestar por el asesinato de Samir Flores y para que se suspendiera la consulta. En particular destaca una entre Huexca y Cuautla por la mañana y otra en la tarde del mismo día en la ciudad de México; del Ángel de la Independencia al Palacio Nacional. En ambas, participaron los colectivos más agraviados, y parientes y amigos del asesinado. El ambiente de la marcha fue de ira, con pancartas, pinta y consignas sobre la traición del presidente de la República, y de ser el asesinato de Samir un crimen de Estado.5 En este escenario de crispación y encono, se celebró la consulta el 23 y 24 de febrero. Como era de esperarse, en las zonas en donde no existían afectaciones directas el resultado fue favorable a la termoeléctrica, considerando que se les prometieron cuotas de electricidad más bajas. En varias localidades de Morelos y Puebla, sin embargo, no se instalaron las casillas o fueron destruidas por pobladores inconformes. El lunes 25 de febrero López Obrador informó los resultados generales de la consulta, en los que el 59.5% voto por el sí a la termoeléctrica y el 40.1% en contra de su culminación. Se emitieron 53,532 votos. A la fecha, en la página de Participación Social de la Secretaría de Gobernación, está todo el diseño y metodología de la consulta, pero no existen los resultados desagregados por casilla (Participación social, 2020). Ante el resultado favorable, se impondría el inicio de las operaciones de la termoeléctrica; no obstante, a la fecha siguen existiendo causas que lo impiden. Son de dos tipos: las tangibles y las intangibles. En las primeras se cuentan las acciones legales en contra del funcionamiento de varios de los componentes del PIM y las recomendaciones de la CNDH (2018), destacando la “Recomendación No. 3/2018”. Existen amparos en los que se suspende temporalmente la operación del gasoducto, en tanto la SENER no realice una consulta a la población indígena por las afectaciones del gasoducto o por los riesgos que hay dado la cercanía del volcán Popocatépetl y su actividad volcánica. Existen también amparos contra el uso de las aguas limpias de los ríos de la región de Cuautla, pues como se dijo, el caudal para la operación sería únicamente con el uso de aguas negras tratadas. Finalmente, también se fincaron amparos específicos en contra del inicio de la operación de la termoeléctrica. A lo anterior se suma otra recomendación de la Comisión Nacional de los Derechos Humanos. El 30 de agosto de 2019, el Consejo de la Judicatura de la Federación emitió un comunicado en el que 5. Se realizó trabajo de campo para seguir el desarrollo de ambos acontecimientos. 49 Alfonso Barquin Cendejas, Mariana García Galicia Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México se concentran en un solo Juzgado, los doce juicios de amparo vigentes a la fecha, relativos a todas las partes del PIM (Lo de hoy Puebla, 2019). Así que, mientras no se desahoguen todos los amparos y no se revoquen los argumentos de los afectados, la termoeléctrica continuará parada. En cuanto a las causas intangibles están claramente la inconformidad popular y las estrategias para impedir el arranque de las operaciones. Como ya se evidenció, existe población organizada en los tres estados y dispuesta a la acción directa en caso de que se inicie el funcionamiento de la termoeléctrica, aun en el caso de que las sentencias le den la razón al gobierno federal. Muchas personas están muy enojadas, lo que es claro por la violencia que se dio en la zona durante la consulta. Se potencia esta ira por el asesinato no resuelto de Samir Flores, que no da muestras de ser aclarado por la apatía y falta de acción de la Fiscalía General del Estado de Morelos. Gran parte de la población de Huexca, como la de Amilcingo de donde era originario Samir, están alertas ante cualquier movimiento, y muestran una alta convicción para evitar la operación de la termoeléctrica. Hay, sin embargo, una conexión dramática que da a los intangibles un peso mayor en esta lucha entre el gobierno federal y los movimientos opositores. El activista Samir Flores, indígena náhuatl, era miembro del Congreso Nacional Indígena (CNI), colectivo indígena de alcance nacional con nexos profundos con el Ejercito Zapatista de Liberación Nacional (EZLN) (Congreso Nacional Indígena, 2019). Este nexo político hace que el asesinato de Samir tome un significado diferente, pues no solo existen actores regionales que se sienten agraviados, indignados e intimidados por el asesinato, sino que grupos de todo el país están adheridos a la causa que representaba Samir y, por lo tanto, significan un apoyo político de una gran fuerza nacional. Además, lo anterior expone la confrontación ya evidente entre el indigenismo del presidente López Obrador, y la visión no gubernamental y no estatalista que sustenta el CNI y el EZLN, que se ha vuelto muy intensa a propósito de otros megaproyectos que el ejecutivo federal pretende llevar a cabo en el país, siendo los más notables por su dimensión y por los probables efectos dañinos a las comunidades indígenas y al medio ambiente: el “Programa para el Desarrollo del Istmo de Tehuantepec” y el “Proyecto de Desarrollo Tren Maya” (Gasparello, 2020). Diversos actores políticos cercanos al CNI y al EZLN también se han sumado a estas declaraciones dando a estos temas vigencia y alcance nacional. De manera particular desataca que en el manifiesto que el EZLN leyó ante sus bases el 31 de diciembre de 2019, en el acto de celebración del vigésimo sexto aniversario de su lucha, el orador principal, el Subcomandante Moisés, expresó duras palabas a propósito de los megaproyectos del gobierno y también su indignación por el asesinato de su compañero Samir Flores: Y con esto del progreso y la modernidad queremos recordar aquí a un compañero del Congreso Nacional Indígena que fue asesinado este año: el hermano y compañero Samir Flores Soberanes. Y lo recordamos porque él se preguntaba y preguntaba que para quién es ese progreso del que tanto hablan. O sea que el hermano Samir preguntaba para dónde va ese camino que llaman “progreso”, que ese nombre lleva como letrero la bestia de los megaproyectos. Y se respondió que ese camino lleva a la destrucción de la naturaleza y a la muerte de las comunidades originarias. Y entonces lo dijo claro que no está de acuerdo, y se organizó con sus compañeras y compañeros, y se resistió, y no tuvo miedo. Y por esa razón es que el mandón que está ahora lo mandó matar. (Enlace Zapatista, 2019) Este tema ocupó en el discurso un lugar especial junto con el de los megaproyectos. Se destacó la lucha de activistas en contra de estos proyectos y de manera más que evidente imputó al mandón “que está ahora” como el responsable del asesinato de Samir. Se hizo también alusión a la manera en que el gobierno federal ha realizado consultas que no son representativas del sentir de la gente y sí de los intereses gubernamentales: “…hace un año, en diciembre del 2018, el capataz que ahora manda en la finca que se llama “México”, hizo una simulación de que le pidió permiso a la madre tierra para destruirla. Entonces se consiguió unas cuantas personas disfrazadas de indígenas y pusieron en la tierra un pollo, trago y tortillas. Así cree el capataz que la madre tierra le da permiso para matarla y hacer un tren que debería llamarse como su familia del capataz. Hace así porque desprecia a los pueblos originarios y porque desprecia a la madre tierra. 50 REFLEXIÓN POLÍTICA AÑO 23 Nº 47 JUNIO DE 2021 ISSN (en línea) 2590-8669; IEP - UNAB (COLOMBIA) Y además el capataz no se quedó ahí, también retó a todos los pueblos originarios y dijo que no le importa lo que pensamos y sentimos, que “les guste o no les guste” a los indígenas, él va a hacer lo que le ordenó su patrón, o sea el mandón, o sea el gran capital.6 (El Universal, 2020) Este choque con los zapatistas del EZLN y su mundo simbólico tiene un capítulo final, que cierra el conflicto que opone las visiones de desarrollo y tradición que encierra el tema de los megaproyectos. Dijo John Womack (2004) en su conocida obra acerca de Zapata y la Revolución Mexicana, que su trabajo era sobre unos campesinos que no querían cambiar, y por eso mismo hicieron una revolución. La batalla en el terreno de las genealogías del poder político y la legitimidad es en este caso muy demostrativa de la situación política vigente, pues en efecto, los inconformes del presente adheridos a la idea del zapatismo original protestan por una obra que supone desarrollo futuro, pero que los desplaza, y afecta la continuidad de su comunidad. En el 2014 López Obrador en terreno zapatista comentó lo siguiente: “Imagínense lo que significa el que en la tierra donde nació Emiliano Zapata, el mejor dirigente que ha habido en la historia de México, aquí, en Anenecuilco, ahí quieren llevar a cabo una termoeléctrica” (El sol de Cuernavaca, 2019). Existía en el candidato un orgullo por la filiación política y un reconocimiento a Zapata como el mejor líder social en la historia del país. Esta convicción se reafirmó, ya que al inicio de su gobierno denominó al año 2019 como el “año de Zapata”, pues se cumplían 100 años de su asesinato; incluso se le incluyó en el logotipo del gobierno de México en los documentos oficiales. Por ello destacó inmediatamente el choque con los manifestantes por la muerte de Samir Flores, con la imputación de ser un crimen de Estado y la igualación simbólica entre Samir y Emiliano Zapata al abundar los gritos de “Samir vive, la lucha sigue” que son una forma muy tradicional de mentar al Caudillo del Sur en toda marcha en la que se invoca su presencia; lo mismo sucede en las pancartas o posters en los que sus rostros aparecen juntos. El enfrentamiento llegó a tal nivel que los inconformes, entre los que se encontraba el nieto de Zapata, ocasionaron que el presidente de México no estuviera presente en la ceremonia de conmemoración del 10 de abril de ese año. Así, nuevamente la población local de Morelos se organizó para evitar el contacto entre el poder gubernamental y Zapata, lo que reitera el contenido simbólico del conocido gesto del héroe al evitar sentarse en la silla presidencial. La intensidad de esta pugna no baja a la fecha, pues en el tradicional mensaje que el EZLN dirigió por el aniversario del inicio de su lucha, en enero de 2021, continúan los mensajes de reclamo por el asesinato de Samir Flores y los nulos avances en la investigación. En el reciente segundo aniversario de su muerte, 20 de febrero de 2021, el EZLN y el CNI, organizaron marchas conmemorativas en los distintos “Caracoles” en la región chiapaneca.7 Como se puede ver, la tensión entre los proyectos de alcance nacional y el rechazo local por los pocos beneficios directos y los daños evidentes queda expuesta de manera evidente en el caso del PIM, con la termoeléctrica como su eje central. El inevitable choque nacion-localidad, impone la necesidad de que estos proyectos sean ser operados políticamente con información suficiente sobre daños y beneficios y desde luego, que las consultas cumplan con los estándares internacionales y nacionales; en particular con lo recomendado por la CNDH en armonía con el Convenio 169 de la Organización Internacional del Trabajo, que reconoce la necesidad de una consulta: libre, previa, informada, culturalmente adecuada y de buena fe (CNDH, 2016).Es evidente que la consulta sobre el PIM no cumplió con dichos requisitos mínimos en su afán por impulsar la termoeléctrica. Tal es el caso en este gobierno que una consulta con mayor tiempo de planeación gubernamental y mayor supervisión ciudadana, la del mencionado “Tren Maya”, no cumplió con dichos estándares, a decir de la Oficina del Alto Comisionado de las Naciones Unidas para los Derechos Humanos (ONU, 2019). Estas consultas se refieren a megaproyectos que el Gobierno mexicano promueve y, por lo tanto, han sido fuertemente cuestionadas porque su diseño beneficia los intereses gubernamentales frente a los intereses locales. Paradójicamente, una nueva consulta sobre un megaproyecto ha sido realizada, 6. Se refiere a una ceremonia ritual el día 15 de diciembre de 2019 en la que personas que se ostentaban como representantes de los 12 pueblos mayas de la región, junto con el presidente de la Republica, ofrendaron bebidas alcohólicas y alimentos a la “Madre Tierra” para pedirle permiso para realizar las obras del “Tren Maya”. El 30 de junio de 2020 el gobierno inició las obras sin que se conozca la Manifestación de Impacto Ambiental. 7. Imágenes disponible en: http://enlacezapatista.ezln.org.mx/2021/02/20/imagenes-de-la-movilizacion-zapatista-en-la-jornada-por-la-vida-contra-losmegaproyectos-y-por-nuestro-hermano-samir-flores/?fbclid=IwAR2MHlF2hdCRYrnVHq1rx1OQs_5b3HlUK1csJSLB-ozNTEpI8epM_uQVoQQ Consultado el 19 de abril de 2021. 51 Alfonso Barquin Cendejas, Mariana García Galicia Tradición contra desarrollo hegemónico, ejes de un conflicto político: El caso del megaproyecto de Huexca en México esta vez para detener su operación. El pasado mes de marzo de 2020, el Gobierno Federal organizó una consulta sobre el inicio del funcionamiento de una planta cervecera de Constellation Brands que tenía todos los permisos necesarios. La consulta fue definida como ilegal ahora por los actores económicos del país, tanto por referirse a una obra casi terminada dentro de los marcos legales, como por carecer de representatividad al votar únicamente el 5% de los ciudadanos de la localidad (Expansión, 2020). Estos tres casos han generado un gran debate en el país, porque enfrentó actores contrapuestos, pero sobre todo, porque evidenció que las consultas oficiales no cumplían todos los estándares de legalidad y equidad, debido a que su diseño y ejecución estuvo sesgado para promover los intereses del gobierno, lo que evidentemente ocasiona grandes tensiones y deja los conflictos sin resolver, como en el caso de Huexca. 5. La división local. Resultados de campo En el año de 2012 se iniciaron los trabajos de construcción de la termoeléctrica sin avisar a la comunidad. Al ser un terreno particular, la CFE lo adquirió de manera privada y comenzó los trabajos sin necesidad de gestiones en Huexca.	12,375
https://openalex.org/W2806970021	OpenAlex	Open Science	CC-By	2,018	Histo-physiologic changes in rat thymus under long exposure to low doses of dichlordiphenyltrichlorethane	Е. П. Тимохина	Russian	Spoken	3,749	10,889	ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 Th e eff ects of diff erent training programs on tra- pezius muscle of women with work-related neck and shoulder myalgia // Acta Neuropathol. (Berl.). — 2000. — Vol. 100. — №3. — P. 253–258. Поступила 13.02.2018 16. Luck J.V.Jr, Andersson G.B.J. Occupational shoulder dis- order, in Rockwood C.A.Jr., Matsen F.A / III (eds): Th e Shoul- der. — Philadelphia: PA, WB Saunders. — 1990. — Vol. 2. — Р. 1088–1108. 16. Luck J.V.Jr, Andersson G.B.J. Occupational shoulder dis- order, in Rockwood C.A.Jr., Matsen F.A / III (eds): Th e Shoul- der. — Philadelphia: PA, WB Saunders. — 1990. — Vol. 2. — Р. 1088–1108. ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 ripheral nervous system in women engaged into construction in- dustry // Industr. med. — 1994. — 4. — Р. 31–33 (in Russian). 13. Suvorov V.G. Syndrome of shoulder rotators compression: clinical signs, diagnosis // Industr. med. — 2008. — 10. — Р. 15–19 (in Russian). force and posture // PREMUS 2007: proc. 6th Intern. sci. conf. prevent. work-relat. musculoskelet. disord. — Boston, 2007. — P. 305. ripheral nervous system in women engaged into construction in- dustry // Industr. med. — 1994. — 4. — Р. 31–33 (in Russian). ripheral nervous system in women engaged into construction in- dustry // Industr. med. — 1994. — 4. — Р. 31–33 (in Russian). 13. Suvorov V.G. Syndrome of shoulder rotators compression: clinical signs, diagnosis // Industr. med. — 2008. — 10. — Р. 15–19 (in Russian). 19. Sommerich C.M., McGlothlin J.D., Marras W.S. Occupa- tional risk-factors associated with soft -tussue disorders of the shoulder — a review of recent investigation in the literature // Ergonomics. — 1993. — Vol. 36. — №6. — P. 697–717. 14. Hagberg M., Harms-Ringdahl K., Nisell R et al. Rehabilita- tion of neck-shoulder pain in women industrial workers: a ran- domized trial com-paring isometric shoulders endurance training with isometric shoulder strength training // Arch. Phys. Reha- bil. — 2000. — Vol. 81. — №8. — P. 1051–1058. g 20. Stone W.E. Repetitive strain injuries // Med. J. Aust. — 1983. — №2. — Р. 616–618. 20. Stone W.E. Repetitive strain injuries // Med. J. Aust. — 1983. — №2. — Р. 616–618. 21. Westgaard R. H., Vasseljen O., Holte K.A. Trapezius muscle activity as a risk indicator for shoulder and neck pain in female service workers with low biome-chanical exposure // Ergonom- ics. — 2001. — Vol. 44. — №3. — P. 339–53. 21. Westgaard R. H., Vasseljen O., Holte K.A. Trapezius muscle activity as a risk indicator for shoulder and neck pain in female service workers with low biome-chanical exposure // Ergonom- ics. — 2001. — Vol. 44. — №3. — P. 339–53. 15. Kadi F. Th e eff ects of diff erent training programs on tra- pezius muscle of women with work-related neck and shoulder myalgia // Acta Neuropathol. (Berl.). — 2000. — Vol. 100. — №3. — P. 253–258. 15. Kadi F. СВЕДЕНИЯ ОБ АВТОРАХ Суворов Вадим Германович (Suvorov V.G.), зав. отд. проф. и непроф. заболеваний ФГБНУ «НИИ МТ», д-р мед. наук. E-mail: margo–183@rambler.ru. 17. McGonagle D., Lories R.J., Tan A.L., Benjamin M. Th e con- cept of a «synovio — entheseal complex» and its implications for understanding joint infl ammation and damage in psoriatic ar- thritis and beyond. Arthritis Rheum, 2007 Aug;56(8):2482–91. 17. McGonagle D., Lories R.J., Tan A.L., Benjamin M. Th e con- cept of a «synovio — entheseal complex» and its implications for understanding joint infl ammation and damage in psoriatic ar- thritis and beyond. Arthritis Rheum, 2007 Aug;56(8):2482–91. Ачкасов Евгений Евгеньевич (Achkacov E.E.), зав. каф. спорт. мед. и мед. реабилитации ФГАОУ ВО «Пер- вый МГМУ им. И.М. Сеченова» Минздрава РФ, д-р мед. наук, проф. E-mail: lfk -cm-mgmy@ya.ru. y g 18. Silverstein B. Work-related rotator cuff and carpal tunnel syndrome quantitative exposure — response relationships with y g 18. Silverstein B. Work-related rotator cuff and carpal tunnel syndrome quantitative exposure — response relationships with y g 18. Silverstein B. Work-related rotator cuff and carpal tunnel syndrome quantitative exposure — response relationships with УДК 591.8+632.15+613.63 Тимохина Е.П., Яглова Н.В., Яглов В.В., Обернихин С.С. Тимохина Е.П., Яглова Н.В., Яглов В.В., Обернихин С.С. ФГБУ «Научно-исследовательский институт морфологии человека» РАМН, ул. Цюрупы, 3, Москва, РФ, 117418 Многие исследователи сходятся во мнении, что одной из при- чин этого можно считать загрязнение окружающей среды [3,7]. К универсальным загрязняющим веще- ствам относится инсектицид широкого действия — ДДТ, который встречается во всех экосистемах мате- риков и океанов, включая Арктику и Антарктиду, и способен длительно сохраняться в почве и воде [1,10]. После продолжительного запрета использование ДДТ в качестве инсектицида возобновилось в 2006 г. со- гласно рекомендации Всемирной организации здраво- охранения по борьбе с переносчиками малярии [15]. В связи с повсеместным распространением ДДТ его содержание в продуктах питания, воде, почве, воздухе регламентировано нормативными документами [2]. Таким образом, изучение влияния фоновых доз ДДТ на организм человека и животных и, в частности, на иммунную систему является актуальной проблемой современной медицины и экологии. Цель исследования — выявление гистофизиоло- гических изменений тимуса крыс при длительном воз- действии низких доз ДДТ. Материал и методики. Эксперимент выпол- нен на 64 самцах крыс Вистар массой тела 80–100 г. Животные содержались в виварии, уход за ними осуществлялся по нормам и правилам обращения с лабораторными животными, в соответствии с «Международными рекомендациями по проведе- нию медико-биологических исследований с исполь- зованием животных» (1985 г.), правилами лабора- торной практики в Российской федерации (приказ МЗ РФ от 19.06.2003 г. №267) и законом «О защите животных от жестокого обращения» гл. V, ст. 10, 4679-ГД от 01.12.1999 г. Эксперимент проведен в соответствии с правилами работы с использовани- ем экспериментальных животных, утвержденными приказом Минздрава СССР № 577 от 12.08.1977 г. Животные опытной группы вместо воды получали растворы о,п-ДДТ («Sigma», США) с концентра- цией 20 мкг/л. Выбор данных доз обусловлен раз- нящимся фоновым содержанием ДДТ на различ- ных географических территориях и максимально допустимым уровнем (МДУ) в продуктах питания (мясная продукция — 0,1 мг/кг; молочная продук- ция — 0,05 мг/кг; зерновые культуры — 0,02 мг/ кг) [2]. По расчетам, потребленная среднесуточная доза ДДТ составила 1,89±0,086 мкг/кг/сут. Жи- вотные контрольной группы получали водопрово- дную воду. Доступ к воде и пище у крыс был сво- бодным. Первую половину животных контрольной и опытной групп выводили из эксперимента через 6 нед., вторую половину — через 10 нед. Прово- дился забор тимуса и крови. После стандартной гистологической проводки с помощью автомати- зированного гистопроцессора («Tissue-Tek VIP 5 Jr», «Hygeco», Франция) изготавливались срезы Результаты исследования и их обсуждение. Ти- мус крыс контрольной группы через 6 недель после начала эксперимента имел типичное дольчатое стро- ение. Снаружи тимус был покрыт соединительнот- канной капсулой, от которой внутрь отходили тон- кие соединительнотканные прослойки, по которым проходили сосуды. ФГБУ «Научно-исследовательский институт морфологии человека» РАМН, ул. Цюрупы, 3, Москва, РФ, 117418 Выявлены гистофизиологические изменения тимуса крыс Вистар, потреблявших дихлордифенилтрихлорэтан (ДДТ) в различных низких дозах в течение 6 и 10 недель. Изучена гистофизиологическая характеристика тимуса, которая показала, что длительное потребление низких доз ДДТ приводит к усилению инволютивных изменений органа за счет усиления гибели тимоцитов и снижения их пролиферативной активности. Установлено, что в меха- низмах гибели тимоцитов под действием низких доз ДДТ задействован, в основном, р53-зависимый путь апоптоза. Ключевые слова: тимус; ДДТ; апоптоз Ключевые слова: тимус; ДДТ; апоптоз Timokhina E.P., Yaglova N.V., Yaglov V.V., Obernikhin S.S. Histo-physiologic changes in rat thymus under long exposure to low doses of dichlordiphenyltrichlorethane. Research Institute of Human Morphology, 3, Tsyurupy str., Moscow, Russian Federation, 117418 Histo-physiologic changes were revealed in thymus of Wistar rats taking dichlordiphenyltrichlorethane in various low doses during 6 and 10 weeks. Studies covered histo-physiologic characteristics of thymus, that proved long intake of low doses of dichlordiphenyltrichlorethane to increase thymus involution due to intense death of thymocytes and their lower proliferative activity. Findings are that mechanisms underlying thymocytes death under exposure to dichlordiphenyltrichlorethane are mostly of p53-dependent apoptosis. p y y p p p p Key words: thymus; dichlordiphenyltrichlorethane; apoptosis 42 ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 тимуса, которые окрашивались гематоксилином и эозином. Изучение гистологических препара- тов проводилось методом световой микроскопии с использованием микроскопа «Leica DM2500» и компьютерной морфометрии с помощью програм- мы «ImageScope» («Leica Microsystems, GMBH», Австрия). В гистологических препаратах тимуса определялись соотношение коркового и мозгового веществ, количество тимических телец на мм2 пло- щади среза. Проводилось иммуногистохимическое исследование экспрессии белка р53 с использова- нием первичных кроличьих поликлональных анти- тел («Santa Cruz Biotechnology», США). Визуали- зация реакции осуществлялась с помощью набора реактивов «UltraVision Detection System» («Termo Scientific», США). Препараты докрашивались гема- токсилином Майера. В сыворотке крови методом твердофазного иммуноферментного анализа с помо- щью коммерческих наборов определялась концен- трация кортикостерона («IBL», Германия). Про- водилось определение пролиферации тимоцитов ex tempore [6] и пролиферативной активности в реак- ции бласттрансформации радиоизотопным методом с использованием 3Н-тимидина. В качестве митогена использовался конканавалин А («Sigma», США). Индекс пролиферации вычислялся по формуле: ИП = Конканавалин А — стимулированная пролифера- ция клеток (имп/мин)/пролиферация клеток в среде (имп/мин). Статистическая обработка проводилась с помощью пакета программ «Statistica» («Statsoft Inc.», США) с использованием параметрических и непараметрических методов. Статистически значи- мыми различия считались при р<0,01. В последние годы в мире отмечен значительный рост числа заболеваний, связанных с нарушениями функционирования иммунной системы [15]. ФГБУ «Научно-исследовательский институт морфологии человека» РАМН, ул. Цюрупы, 3, Москва, РФ, 117418 Паренхима каждой дольки была образована лимфоидной тканью, строма сформиро- вана ретикулярными эпителиоцитами. В дольках четко различались две зоны: корковое вещество, занимаю- щее периферическую часть дольки, и мозговое веще- ство, находящееся в центре дольки. Доля коркового вещества составляла около 75% паренхимы (табл.). В корковом веществе лимфоциты плотно прилежали друг к другу. В мозговом веществе встречались еди- ничные тимические тельца (табл.). Экспрессия белка р53 выявлялась приблизительно в 10% клеток тимуса (табл.). р53-позитивные клетки обнаруживались как в субкапсулярном слое, так и на границе коркового и мозгового вещества. Пролиферативная активность ex tempore клеток тимуса крыс контрольной группы через 6 недель после начала эксперимента составила 9025,35±274,00 имп/мин. После введения митогена пролиферативная активность тимоцитов увеличива- лась в 3 раза. 43 ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 Таблица Морфофункциональные характеристики тимуса и изменения концентрации кортикостерона в сыво- ротке крови крыс, потреблявших ДДТ в дозе 1,89±0,086 мкг/кг/сут. в течение 6 и 10 недель и крыс контрольных групп (M±m) Показатель 6 недель 10 недель Контроль- ная группа ДДТ 1,89±0,086 мкг/кг/сут. Контроль- ная группа ДДТ 1,89±0,086 мкг/кг/сут. Доля коркового вещества, % 73,37±1,40 75,49±0,89 76,4±1,00 74,98±1,37 Количество тимических телец на мм2 0,8±0,15 1,61±0,30* 1,84±0,29 2,42±0,47# Тимоциты, экспрессирующие белок р53, % 9,20±0,03 20,43±0,01 20,98±0,03 20,24±0,01 Концентрация кортикостерона в сыворотке крови, нг/мл 139,09±10,99 108,41±1,94* 127,12±8,99 108,09±2,75* Примечание: * — статистически значимое отличие от контрольной группы; # — статистически значимое отличие от опыт- ной группы меньшего срока исследования. Таблица Морфофункциональные характеристики тимуса и изменения концентрации кортикостерона в сыво- ротке крови крыс, потреблявших ДДТ в дозе 1,89±0,086 мкг/кг/сут. в течение 6 и 10 недель и крыс контрольных групп (M±m) Через 10 недель потребления ДДТ в дозе 1,89±0,086 мкг/кг/сут. изменений в строении капсулы и междо- левых перегородок в сравнении с предыдущим сро- ком исследования не наблюдалось. Доля коркового вещества не отличалась от показателей контрольной группы, а также группы, потреблявшей ДДТ в этой же дозе в течение 6 недель. Процентное содержание клеток, экспрессирующих белок р53, не изменилось по сравнению с контрольной группой (табл.). р53- позитивные клетки располагались диффузно в корко- вом, а также группами в мозговом веществе. Количе- ство тимических телец в мозговом веществе тимуса крыс увеличилось по сравнению с предыдущим сроком исследования. Было отмечено снижение пролифера- ции ex tempore тимоцитов и повышение пролифера- тивного ответа на митоген по сравнению с контроль- ными значениями. После потребления ДДТ в дозе 1,89±0,086 мкг/кг/ сут. в течение 6 недель при гистологическом исследо- вании тимус имел также типичное строение. ФГБУ «Научно-исследовательский институт морфологии человека» РАМН, ул. Цюрупы, 3, Москва, РФ, 117418 Толщи- на соединительнотканных междолевых перегородок не изменялась, но отмечалось увеличение количества жировых клеток в междолевых перегородках по срав- нению с контрольной группой. Доля коркового и мозгового веществ не отличалась, но по сравнению с контрольной группой, граница между корковым и мозговым веществом была менее четкой. Процентное содержание клеток, экспрессирующих белок р53, в два раза превышало соответствующие значения контроль- ной группы (табл.). р53-позитивные клетки распола- гались диффузно в корковом веществе, а также встре- чались и в мозговом веществе. В мозговом веществе наблюдалось увеличение количества тимических телец (табл.). Исследование пролиферативной активности клеток тимуса крыс показало статистически значи- мое усиление пролиферации ex tempore тимоцитов по сравнению с контрольной группой, однако пролифе- ративный ответ на введение митогена не изменялся. Установлено статистически значимое снижение кон- центрации кортикостерона в сыворотке крови крыс по сравнению со значениями контрольной группы аналогичного срока исследования. При изучении гистологических препаратов ти- муса крыс всех опытных групп в первую очередь была отмечена гибель тимоцитов, проявляющаяся наличием участков опустошения коркового веще- ства. Известно, что гибель клеток тимуса в процес- сах негативной селекции происходит, в основном, за счет апоптоза. По данным работ О. Tebourbi с соавт. воздействие больших доз ДДТ иници- ирует гибель тимоцитов также путем апоптоза [14]. Существуют два механизма апоптоза тимо- цитов — р53-зависимый и р53-независимый пути. р53-независимый путь индуцируется в большин- стве случаев гормонами надпочечников — глюко- кортикоидами [9]. В связи с этим, для установле- ния механизмов запуска апоптоза тимоцитов при действии на них низких доз ДДТ было проведено определение концентрации кортикостерона в сы- воротке крови, являющегося основным глюкокор- тикоидом у крыс. В исследовании во всех опытных группах отмечено снижение уровня кортикосте- рона по сравнению с контрольными значениями. Этот факт позволяет утверждать, что в тимусе крыс под действием ДДТ задействован неглюкокортико- ид-индуцируемый путь апоптоза. Строение тимуса крыс контрольной группы че- рез 10 недель после начала эксперимента не имело существенных различий по сравнению с предыду- щим сроком исследования. Соотношение коркового и мозгового вещества не изменялось. Заметных изме- нений толщины соединительнотканных междолевых перегородок замечено не было, но количество клеток, экспрессирующих белок р53, увеличилось более чем в два раза (табл.). При этом р53-позитивные клетки рас- полагались в основном субкапсулярно. В мозговом ве- ществе отмечалось увеличение количества тимических телец в единице площади среза по сравнению со зна- чениями предыдущего срока исследования. Значения показателей пролиферации тимоцитов в контрольной группе через 10 недель не отличались от предыдущего срока исследования. 44 ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 Потребление ДДТ крысами в дозе 1,89±0,086 мкг/ кг/сут. Выводы: Через 10 недель после начала эксперимента в ти- мусе крыс наблюдались отличия, связанные как с воз- действием ДДТ, так и с возрастными изменениями. У животных контрольной группы отмечалось усиление гибели лимфоцитов, о чем свидетельствует двукратное увеличение доли р53-позитивных клеток и появление в корковом веществе участков гибели лимфоцитов, а также увеличение количества тимических телец в мозговом веществе. Эти факты говорят о начале воз- растной инволюции тимуса у крыс, которая развива- ется после наступления периода полового созревания [11–13]. 1. Длительное воздействие низких доз ДДТ при- водит к изменениям гистофизиологии тимуса, за- ключающимся в ускорении развития инволютивных изменений органа, главным образом за счет усиления р53-зависимого апоптоза тимоцитов и снижения их пролиферативной активности, что, в свою очередь, может быть причиной нарушения реакций клеточно- го иммунитета. у 2. Выявленные изменения тимуса крыс при дли- тельном воздействии низких доз ДДТ показывают, что максимальные допустимые уровни его содержа- ния в продуктах питания не являются безопасными для организма. у 2. Выявленные изменения тимуса крыс при дли- тельном воздействии низких доз ДДТ показывают, что максимальные допустимые уровни его содержа- ния в продуктах питания не являются безопасными для организма. В отличии от контрольной группы, где апоптозу подвергались в основном лимфобласты субкапсу- лярного слоя, в группе, потреблявшей ДДТ в дозе 1,89±0,086 мкг/кг/сут. в течение 10 недель, отме- чался апоптоз лимфоцитов, что и привело к увели- чению очагов опустошения коркового вещества. Так как процентное содержание р53-позитивных клеток не отличалось от значений контрольной группы, то это может объясняться значительной гибелью тимоцитов на более раннем сроке. Было отмечено также усиление гибели ретикулярных эпителиоцитов в этой группе. Снижение темпов пролиферации тимоцитов подтверждается также и усилением их пролиферативной активности при стимуляции митогеном. СПИСОК ЛИТЕРАТУРЫ (см. REFERENCES пп. 10–15) 1. Гигиенические критерии состояния окружающей сре- ды. ДДТ и его производные. 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Изменения ци- токинового профиля у крыс Вистар при длительном воздей- ствии низких доз ДДТ // Иммунология. — 2012. — №2. — С. 98–102. 9. Ярилин А.А. // Иммунология. — 1996. — №6. — С. 10–23. 9. Ярилин А.А. // Иммунология. — 1996. — №6. — С. 10–23. ФГБУ «Научно-исследовательский институт морфологии человека» РАМН, ул. Цюрупы, 3, Москва, РФ, 117418 в течение 6 недель приводило к гибели лимфо- цитов, которая выражалась в двукратном увеличении по сравнению с контрольной группой процентного со- держания клеток, экспрессирующих белок р53. Таким образом, ДДТ даже в низкой дозе способен усиливать гибель клеток в тимусе преимущественно р–53 зави- симым путем апоптоза. Также было отмечено увеличе- ние числа тимических телец, что говорит об усилении гибели ретикулярных эпителиоцитов. Было отмечено статистически значимое усиление пролиферации ex tempore тимоцитов по сравнению с контрольной груп- пой, при этом пролиферативный ответ на введение митогена не снижался. Однако изучение концентрации кортикостерона в данном исследовании показало снижение его уровня во всех опытных группах, а изменения, наблюдав- шиеся через 6 недель воздействия низких доз ДДТ, такие как повышение пролиферативной активности, носили реактивный характер. Более длительное воз- действие инсектицида не вызывало изменений гисто- физиологии органа, направленных на восстановление баланса между гибелью и пролиферацией лимфоци- тов. Это подтверждается и ранее выявленными из- менениями концентрации пролиферативных и ан- типролиферативных цитокинов в сыворотке крови крыс при длительном воздействии аналогичных доз ДДТ [8]. REFERENCES 1. Hygienic criteria of environmental state. Dichlordiphenyl- trichlorethane and its derivatives. Associated publication of UN 45 ISSN 1026-9428. Медицина труда и промышленная экология, № 4, 2018 program on environment and WHO. — Moscow: Meditsina, 1982 (in Russian). program on environment and WHO. — Moscow: Meditsina, 1982 (in Russian). 13. Shanley D., Aw D., Manley N., Palmer D. // Trends in im- munology. — 2009. — Vol. 30. — №7. — Р. 374–381. gy 14. Tebourbi O., Rhouma K., Sakly M. // Bull. Environ. Con- tam. Toxicol. — 1998. — Vol. 61. — Р. 216–223. 2. Hygienic norms of pesticides content of environmental ob- jects (list). — Moscow: Minzdrav RF, 1997 (in Russian). 15. WHO gives indoor use of DDT a clean bill of health for controlling malaria // WHO. — 2008. 3. Maystrova I.N. // Zdravookhr. — 1998. — 8. — Р. 21–23 (in Russian). 4. Basics of human physiology. — St-Petersburg, 1994 (in Russian). Поступила 14.01.2015 5. Essays on adaptational syndrome. Moscow: Moscow: Medg- iz, 1960 (in Russian). СВЕДЕНИЯ ОБ АВТОРАХ 6. Rakhmilevich A.L., Obernikhin S.S. // Immunologiya. — 6. Rakhmilevich A.L., Obernikhin S.S. // Immunologiya. — 1990. — 1. — Р. 68–69 (in Russian). Тимохина Екатерина Петровна (Timokhina E.P.), р р ( ), мл. науч. сотр. лаб. развития эндокринной системы ФГБУ «НИИМЧ» РАМН. E-mail: rodich_k@mail.ru. 7. Frolov V.M. Zhurn. mikrobiologii, epidemiologii i immuno- biologii, 1995; 2: 119–124 (in Russian). 8. Yaglova N.V., Rodichenko E.P., Yaglov V.V. Changes of the cyto- kine profi le in Wistar rats under the long-term action of low DDT doses // Immunologiya. — 2012. — 2. — Р. 98–102 (in Russian). Яглова Наталья Валентиновна (Yaglova N.V.), зав. лаб. развития эндокринной системы ФГБУ «НИ- ИМЧ» РАМН, д-р мед. наук. E-mail: yaglova@mail.ru. g y 9. Yarilin A.A. // Immunologiya. — 1996. — 6. — Р. 10–23 (in Russian). Яглов Валентин Васильевич (Yaglov V.V.), гл. науч. сотр. лаб. развития эндокринной системы ФГБУ «НИИМЧ» РАМН, д-р мед. наук, проф. E-mail: vyaglov@ mail.ru. 10. Daly G., Wania F. // Environ. Sci. Technol. — 2005. — Vol. 39. — №. 2. — P. 385–398. 11. French R., Broussard S., Meier W. // Endocrinology. — 2002. — Vol. 143. — Р. 690–699. Обернихин Сергей Станиславович (Obernikhin S.S.), Ст. науч. сотр. лаб. клеточной иммунопатологии и био- технологии ФГБУ «НИИМЧ» РАМН, канд. мед. наук. E-mail: ober@mail.ru. 12. Greenstein B., Fitzpatrick F., Adcock I., et al. // Endocri- nol. — 1986. — Vol. 110. — Р. 417–422. УДК 616. 127:599.325 Воробьева В.В. 1, Шабанов П.Д. 2 1ФГБОУ ВО «Смоленский государственный медицинский университет» Минздрава России, ул. Крупской, 28, Смоленск, РФ, 214019 РФ, 214019 2ФГБВОУ ВПО «Военно-медицинская академии им. С.М. Кирова» МО РФ, ул. акад. Лебедева, 6, Санкт-Петербург, РФ, 194044 , 2ФГБВОУ ВПО «Военно-медицинская академии им. С.М. Кирова» МО РФ, ул. акад. Лебедева, 6, Санкт-Петербург, РФ, 194044 Проводилось экспериментальное изучение защитных свойств сукцинатсодержащего антигипоксанта реамберина при остром воздействии холода, вибрации и иммобилизации у крыс. Крысы контрольной группы и группы, получавшей реамберин в дозе 50 мг/кг, подвергались воздействию холода (–15 ºС на 60 мин) и иммобилизации после сеанса общей вибрации с амплитудой 0,5 мм и частотой 44 Гц на про- тяжении 90 мин. Оценивались показатели ректальной температуры и сукцинатдегидрогеназной активности лимфо- цитов периферической крови (СДГ-активность) по данным количественного цитохимического теста, основанного на реакции восстановления пара-нитрофиолетового тетразолия до формазана. р р р ф р ф р Ключевые слова: охлаждение; вибрация; иммобилизация; ректальная температура; сукцинатдегидрогеназная активность лимфоцитов периферической крови крыс; янтарная кислота; реамберин Vorobieva V.V. 1, Shabanov P.D. 2 Protective properties of reamberine in acute experimental combined exposure to cold, vibration and immobilization. 1Smolensk State Medical University, Smolensk, 28, Krupskaya str., Russian Federation, 214019; 2Military Medical Academy, 6, Acad. Lebedeva str., St. Petersburg, Russian Federation, 194044 Experimental study covered protective properties of succinate-containing ant-hypoxant reamberine in acute exposure to cold, vibration and immobilization in rats. 46	26,303
W4367595340.txt_1	Open-Science-Pile	Open Science	Various open science	2,023	Artificial neural network–modified heat transfer model for furnace coil rolling of magnesium alloy plate	None	English	Spoken	7,162	11,410	Arti cial neural network modi ed heat transfer model for furnace coil rolling of magnesium alloy plate Shuping Kong Fangkun Ning (  kingfning@126.com ) Northeastern University Key Laboratory of Electromagnetic Processing of Materials Ministry of Education Lifeng Ma Weitao Jia Qichi Le Research Article Keywords: Magnesium alloy, Furnace coil rolling, Heat transfer model, Arti cial neural network Posted Date: October 18th, 2022 DOI: https://doi.org/10.21203/rs.3.rs-2154059/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Artificial neural network modified heat transfer model for furnace coil rolling of magnesium alloy plate Shuping Kong a, b, Fangkun Ning a, b, , Lifeng Ma a, b,, Weitao Jia a, b,, Qichi Le c a b Heavy Machinery Engineering Research Center (MOE), Taiyuan University of Science and Technology, Taiyuan 030024, China College of Mechanical Engineering, Taiyuan University of Science and Technology, Taiyuan 030024, China c Key Laboratory of Electromagnetic Processing of Materials (MOE), Northeastern University, Shenyang 110819, China *Corresponding authors, E-mail addresses: malifengfqh@163.com (L. Ma), kingfning@126.com (F. Ning) Abstract Through the experimental temperature measurement and numerical simulation of veneer and stacked plates with different thicknesses, the heat transfer behavior of magnesium alloy wide plate furnace coil rolling was studied. It found that the radial thermal radiation heat transfer rate of the magnesium alloy coil was greater than the thermal conduction heat transfer rate between the coil layers, and the average heating rate of the outer layer of the coil was about twice that of the middle and inner layers. A dimensionless correction factor λ was introduced to the traditional heat transfer model of magnesium alloy coil rolling process based on artificial neural network (ANN) model. After the modification, the reduced Chi-Spr of the heat transfer models of the coil rolling process was reduced significantly from 6.9195 to 1 nearby. In addition, there was a negative correlation between the plate thickness and the heating rate, and the heating rate of the sheet with a small Biot (Bi) value to different temperatures varied greatly. The heating rate of the 3.6mm sheet augments with the increase of temperature. The cooling time of the magnesium plate was proportional to the thickness of the plate, and the cooling rate of the plate with a large Bi value varied greatly from distinct initial temperatures. The cooling rate of 6.0mm plate at high temperature was greater than that at low temperature. According to the temperature distinction of different parts of the sheet, the precise cooling model was constructed. Keywords: Magnesium alloy; Furnace coil rolling; Heat transfer model; Artificial neural network 1. Introduction Magnesium (Mg) alloys have been widely used in the fields of railway transportation, consumer electronics and aerospace because of their excellent properties such as low density, high specific strength, brilliant thermal conductivity and easy to recycle[1,2]. Further, Mg alloys thick plates can be used in the manufacture of satellite and spacecraft bases, missiles and rockets instrument bulkhead, missiles and aircraft tails[3–5]. Mg alloys medium plates are used to manufacture drop tanks, liner plates and other structural parts of fighter aircraft[6,7]. Wide Mg plates can be used in car doors and front/rear cover plates. Due to the high thermal conductivity and temperature sensitivity of Mg alloy, it is easy to lose heat during rolling deformation, which will lead to uneven temperature distribution of the plate along width direction[8]. Temperature, as one of the important factors affecting the rolling process of Mg alloys, has great influence on the microstructure change, deformation resistance and plasticity of Mg alloys. The ductility of the alloy increases significantly at high temperatures due to dynamic recovery, continuous dynamic recrystallization, grain boundary slip and activation of additional slip systems. The cracks are occurred on the edge when the plate is further rolled, which leads to the decrease of the yield of the Mg plate and the increase of the cost[9–11]. The deformation reduction and rolling speed are also important to the production of Mg alloy plate except for temperature in the rolling process. Excessive deformation is easy to cause stress concentration and lead to plate cracking, while too small deformation will affect production efficiency and product quality. Kim et al. [12,13] constructed a mathematical heat transfer model to predict the heat flux on the slab surface and temperature distribution by considering the thermal radiation in the furnace chamber and transient heat conduction. Guan et al. [14] established the temperature field model of the rolled plate in the rolling process of the Steckel mill based on the analysis of the heat transfer characteristics of the rolled plate in each stage of the rolling zone. Li et al. [15] analyzed the heat transfer and derived the expression of the convective coefficient to the temperature, and the radial thermal conductivity of the aluminum coil has been resolved indirectly applying the golden section method, thus a numerical model for the aluminum coil’s annealing temperature field is established． The hot coil box in the Steckel mill system is an intermediate thermal insulation process equipment installed between roughing mill and finishing mill. The coiling process reduces the heat dissipation surface area of the intermediate billet and the temperature difference between the head and the tail. The change of temperature field of medium thick plate during its stay in the hot-rolled box is accurately mastered to provide a theoretical basis for controlling the fine rolling temperature, improve the quality of Mg plate and reduce energy consumption. However, due to the complex nonlinear dependence of the overall temperature of the coil on the thickness, oxide layer, gas flow, coiling time, etc., it cannot be detected by existing technical means, and it is more difficult to use accurate mathematical models to describe the temperature. Deep learning tools such as artificial neural network (ANN) have shown their ability to handle nonlinear material property predictions that are difficult to express in simple formulas. ANN method has been successfully applied in the field of materials science, such as material design, properties and performance prediction, etc[16–18]. Studies have shown that neural network model has better predictability of heat treatment furnace sheet metal temperature than traditional heat transfer model[19–21]. Li et al.[19] improved the temperature prediction accuracy of the heat treatment furnace sheet based on the automatic optimization of structural parameters and weight thresholds of the differential neural network. Zhang et al.[21] proposed a control strategy for furnace temperature based on machine learning intelligent control of industrial furnaces, and built a combustion control model and a machine learning model, which can make the furnace temperature respond in time and accurately control the heat treatment process requirements of products. As far as we know, a series of complex functional relations such as differential equations were used in the process of establishing the neural network model in the previous work. In the study of heat transfer in furnace coil rolling, it is better to directly establish a neural network model between various parameters. The ANN method is used to introduce a dimensionless correction factor λ to correct the heat transfer model of coil rolling. 2. Experimental procedure and model 2.1 Model and method Fig. 1(a) shows the schematic diagram of Steckel mill rolling. After the slab came out of the heating furnace, it is transported to the Steckel mill through the transfer roller for rough rolling. The rolled plate is then put into the hot coil box for heat preservation and is ready for reciprocating rolling. After each pass, it entered the hot coil box to keep warm until the thickness of the rolled plate reached 7~13mm. With this thickness of magnesium alloy coil as the research object, heat transfer and thermal stress analysis are carried out to reduce the unevenness of the structure and performance of the coil, and to provide high-quality coils before finishing rolling. Fig. 1(b) and (c) are the schematic diagrams of Mg alloy coil and layered simulation geometry model, respectively. Where AD is the axis direction, RD is the radial direction, and TD is the tangential direction. The thermal conductivity of the coil in the AD is equal to the thermal conductivity of the material, and the thermal conductivity in the RD is smaller than that in the AD due to the gap between the multilayer materials. In order to modify the heat transfer model of coil rolling, a dimensionless correction factor λ is introduced using the ANN method. The entire learning process is based on an error compensation algorithm. Fig. 2 is schematic diagram of coil temperature control strategy. Fig. 1 Schematic diagrams of Steckel mill rolling (a), Mg alloy coil (b) and layered simulation geometry model (c) Feedback Compensation Actual Furnace Tem. Heating Power Insert Gas Pressure Heating Model & Cooling Model Heating System Control Mechanism Calculation Into-furnace Tem. Machine Learning Ambient Tem. Coil Thickness Coiling Time Biot Value Coil Width Furnace Temperature Prediction Model Forecast Error Compensation Insert Gas Flow Coil Temperature Uniformity Coil Temperature Prediction Model Coil Mass Fig. 2 Schematic diagram of coil temperature control strategy 2.2 Numerical procedures The thermal radiation, thermal conduction and thermal stress changes of the AZ31 magnesium alloy coil in the hot coil box are simulated in the ABAQUS software using a thermo-mechanical coupling reduction integration unit (C3D8RT). The basic assumptions of the model are as follows, ① The material is an isotropic continuous solid; ② The gas temperature in the hot coil box is kept at a constant temperature; ③ The time interval between the coil entering the hot coil box from the head to the tail entering depends on the thickness of the coil; ④ Only the influence of temperature on stress is considered, and the influence of factors such as structure change and deformation heat are not considered ; ⑤ The axial thermal conductivity of the coil is the same as the thermal conductivity of the material. The size of the coil is Φ 600mm×Φ1200mm×1500mm, according to the geometric symmetry of the simulation object and the symmetry of the boundary conditions, take 1/8 of it for modeling, as shown in Figure 1c. The thickness of the plate is 6.0mm, and the coil has a total of 23 layers. The temperature of the inner layer is 1°C higher than the temperature of the outer layer, the temperature of the outermost layer is set to 200°C, and the temperature in the hot coil box is 250°C. After a certain period, the influence of the thermal resistance between the coil layers and the holding time on the temperature field and stress field of the coil is analyzed. 3. Results and discussion Heating and cooling experiments with different thicknesses and gaps between layers are conducted. Experiment and simulation cooling curves of different plates are shown in Fig. 3. Fig. 3a showed the cooling curve of the plate from the initial temperature of 350 °C, and Fig. 3b is the cooling curve of the plate from the initial temperature of 200 °C. It could be seen from Fig. 3 that the smaller the thickness of the plate, the faster the temperature drop of the plate, but the greater the temperature difference between the middle and the edge of the plate. The measured curve and the simulated curve of the veneer cooling are in good agreement when the thermal convection heat transfer coefficient h is 0.01N/s·mm·K, but the stacked plates are affected by the heat radiation, convection, and contact heat conduction, so the temperature is lowered. The above factors should be considered in the simulation curve. Fig. 3 Experiment (a) and simulation (b) cooling curves of different plates 3.1 Effect of thermal radiation on coil temperature Fig. 4 showed the actual furnace coil mill and the emissivity at different temperatures. After the slab came out of the coiling furnace, it passed through the front pinch rolls and entered the six-high reversing mill for rolling, and the thickness, flatness and tension of the plate are tested at the same time, and finally entered the coiling furnace to keep warm. The temperature of the plate changed continuously throughout the process, and the emissivity varied substantially linearly with different temperature as shown in Fig. 4b. (a) (b) Fig. 4 Furnace rolling mill and emissivity with different temperatures The thermal insulation process of the coil in the coiling furnace is simulated, as shown in Fig. 5. Since the head of the plate first entered the coiling furnace, the temperature of the inner layer of the coil is higher than that of the outermost layer. The temperature of the innermost layer is set to 223°C, the temperature of the outermost layer is 200°C, and the temperature in the furnace is 250°C. It could be seen from Fig. 5a that the temperature of the inner layer coil reached 250°C after 500s, the center layer coil got to 250°C after about 700s (Fig. 5b), and the outer layer coil warmed 250°C after 495s (Fig. 5c). The coils of the inner and outer layers heated up faster due to the heat radiation of the air in the furnace in the axial and radial directions, while the temperature of the center layer coils raised slowly because of the heat radiation in the two axial directions and the heat conduction of the adjacent layers. It could be seen from Fig. 5 that the thermal radiation heating effect of the entire coil axis is greater than that of the radial direction, which could provide a reference for the arrangement of hot air in the coiling furnace. (a) (b) (c) (d) Fig. 5 Heating process of the coil in the coiling furnace (a) Intrados; (b) Inner layer; (c) Extrados; (d) Whole coil The temperature change during the heating process is recorded at the middle position of the roll axis of each layer (Fig. 6). It could be seen from Fig. 6 that the temperature of the intermediate layer coil decreased after the initial stage (within 20s) and then raised. The reason is that the intermediate layer is in contact with the lower temperature coil on the outside, resulting in a temperature drop. Due to the low initial temperature of the outermost coil, the temperature rise rate is higher than that of the innermost coil, but the two layers took the same time to reach the target temperature, and the middle layer is delayed by about 200s. When the coil is coiled in the coiling furnace, there are two ways to keep the coil temperature constant. One is to increase the heating rate of the intermediate layer, and the other is to make the coil temperature consistent before entering the coiling furnace. Otherwise, it is easy to cause uneven grain microstructure in different parts of the coil, and it is difficult to guarantee the accuracy of the plate in the subsequent finishing rolling process. Fig. 6 Heating curve at the center of different layers Combined with the actual temperature measurement of the plate, Fig. 7 and Fig. 8 are the temperature-time curves of the same size and thickness of 3.6mm and 6.0mm plates heated to 200°C and 350°C respectively. As can be seen from Fig. 6a, it took 1630s for the temperature of the 3.6mm thick plate to rise from 50°C to 200°C, while in Fig. 7b, it took 1900s for the temperature of the 6.0mm thick plate to increase from 50°C to 200°C. As can be seen from Fig. 8, it took 2050s and 3944s for 3.6mm and 6.0mm thick plates to rise from 50°C to 350°C, respectively. The heating rates of plates with different thicknesses are shown in Table 1. It could be seen from Table 1 that the average temperature rise rate of 3.6mm to 200°C is 0.0920°C/s, while the average temperature rise rate of 6.0mm to 200°C is 0.0789°C/s. The average heating rates of 3.6mm and 6.0mm plates up to 350°C are 0.1463°C/s and 0.0761°C/s, respectively. The heating rate of 6.0mm thick plate to 200℃ and 350℃ is not much different, while the rate of 3.6mm thick plate heating to 350℃ is 59% higher than that of heating to 200℃. From the knowledge of heat transfer, the temperature distribution in the thickness direction of the slab can be quantitatively characterized by the Biot number (Bi). As the thickness of the plate increases, the heating effect of the air will be affected by more thermal resistance of the material, which will reduce the ability of the heating effect to penetrate the slab, thereby reducing the heating rate of the thick to the middle layer. With the increase of the initial temperature, the temperature gradient in the thickness direction gradually increases. This is because the heat radiation capacity increases in the high temperature stage after the plate temperature rises, resulting in a sharp temperature rise of the surface layer per unit time. The smaller the Bi, the smaller the internal thermal resistance and the greater the external thermal resistance. When participating in engineering calculations, Bi≈0.1 means that the temperature distribution of the entire object is relatively uneven, and the larger the Bi, the more significant the temperature unevenness. Table 1 Heating rate of different plate thickness Thickness/mm Rate of heating up to 200°C/(℃/s) Rate of heating up to 350℃/(℃/s) 3.6 0.0920 0.1463 6.0 0.0789 0.0761 (b) (a) Fig. 7 Heating-cooling curves of 3.6mm and 6.0mm thickness plates (a) 3.6mm; (b) 6.0mm (a) (b) Fig. 8 Curves of heating up to 350°C for 3.6mm and 6.0mm thickness plates 3.2 Effect of convection heat transfer coefficient on coil temperature 3.2.1 Time-history curves correlated with temperature changes Fig. 7 and 8 are the temperature-time curves of the same size and thickness of 3.6mm and 6.0mm plates heated to 200°C and 350°C, respectively. It can be seen from Fig. 7a that it takes 540s for the temperature of the 3.6mm thick plate to cool down from 200°C to 50°C by air cooling. It can be seen from Figure 7b that it takes 800s for the temperature of the 6.0mm thick plate to cool down from 200°C to 50°C by air cooling. As the thickness of the sheet increases, the thermal resistance of the sheet increases, and the heating and cooling rates of the sheet increase accordingly. For coils, a small amount of air should be kept warm when entering the coiling furnace, and the air volume will increase as the coiling proceeds. It could be known from the literature [22] that during the cooling process, the ratio of the heat flux density of convection heat transfer in the furnace to the heat flux density of radiative heat transfer in the cooling process could reach several hundreds of times. Currently, the temperature of the coil and the furnace is shallow, and the radiation heat transfer conditions are inferior. In the soaking stage, the temperature of the magnesium coil and the furnace is high, and the above ratio is just ten times. Therefore, it could be seen that the convection heat transfer played a decisive role in the heat transfer in the coiling furnace. The use of inert gas with a density lower than that of air could increase the diameter of the atmosphere circulating fan and greatly augment the amount of air circulation, thereby aggrandizing the convection heat transfer coefficient in the furnace. Fig. 9 shows the combination of different convective heat transfer coefficient h values for magnesium plates with a thickness of 3.6 mm and 6.0 mm. It could be seen from Fig. 9 that with the increase of h value, the cooling rate of the sheet augments, but the temperature difference of the 3.6mm sheet is greater than that of the 6.0mm plate under the same h value in the width direction, which further showed that the heat dissipation rate of the sheet in the width direction is faster, and manifesting the cooling curves for the same h value are scattered. The fitting values of each cooling curve are shown in Table 2. The time took for each h value to drop from 200 °C to 50 °C is shown in Table 2. It is found by calculation that under the same conditions as thermal emissivity, convection heat transfer coefficient, starting and ending temperature, etc., the cooling time of different plate thicknesses is proportional to the plate thickness (Fig. 10), the slope is 1.5547 and the ratio of plate thickness is almost equal. The thickness of the plate also affects the thermal conductivity. The radial thermal conductivity varies with the thickness of the magnesium coil and the thickness of the gap between the magnesium coils. The thicker the magnesium plate, the greater the thermal conductivity, and the greater the equivalent thermal conductivity. Table 2 Cooling simulation curve of initial temperature 350℃ h Cooling 3.6mm thickness--ε0.15 0.008 T = 26.9632 + 316.0900 exp(− 0.010 T = 23.4170 + 319.6683exp(− 0.015 T = 21.4902 + 322.8523exp(− 0.020 T = 20.5614 + 324.9434 exp(− time  351.5396  284.5925  210.5770  162.9282 Cooling 6.0mm thickness-ε0.15 ) 700s T = 36.7066 + 307.4442 exp(− ) 540s T = 29.4189 + 313.9883exp(− ) 385s T = 22.4346 + 320.3219 exp(− ) 305s T = 20.7385 + 322.1106 exp(− time  507.1481 ) 1075s ) 850s ) 610s ) 455s  442.6721  326.5254  254.4572 (b) (a) Fig. 9 Cooling curves of different h combinations of 3.6mm and 6.0mm magnesium plates (a) 3.6mm; (b) 6.0mm Fig. 10 Ratio of cooling time for different plate thicknesses 3.2.2 Quantitative analysis about cooling rate The experimental and simulated cooling curves are fitted under different conditions, and it is found that they are all single-exponential decay cooling curve equation. Table 3 Cooling curve equations of distinct plates (Experiment) Category Equation form H5.6-200C (Experiment) T = 28.5114 + 171.9322 exp(− H3.6-200C (Experiment) T = 27.3496 + 174.8338exp( − H3.6-350C (Experiment) H5.6-350C (Experiment) H3.6+5.6-350C (Experiment) Reduced Chi-Sqr  ) 2.3862 ) 2.6251 418.8544  279.2653  ) 314.5478  T = 43.1557 + 306.7860exp( − ) 431.3252  T = 42.4033 + 307.4190 exp(− ) 694.8143 T = 37.0228 + 323.9742 exp( − 6.9195 3.6076 16.7111 H3.6+5.6-350C (Simulation) h=0.01N/s·mm·K H3.6+5.6-350C (Simulation) h=0.008N/s·mm·K  ) 851.3863  T = −19.1601 + 373.4150 exp( − ) 1075.9468 T = −11.3874 + 364.3091exp(− 0.9819 1.7621 The parameters of the magnesium veneer cooling curves with different thicknesses and initial temperatures are fitted, and then brought into the furnace heat transfer equation to obtain the general form of cooling in the magnesium plate coil furnace. 3.3 ANN-modified temperature control model establishment 3.3.1 Rolls transportation progress Rolls transportation is a necessary link in the rolling preparation of magnesium alloy wide coil. The slab dissipates heat in the air by means of internal heat conduction and external heat conduction (convection and radiation) without auxiliary heat, resulting in temperature drop in slab. The accurate prediction of the temperature of magnesium plate in the conveying process can provide basic data and models for the implementation of temperature control and heat supplement in the whole rolling process. The heat transfer modes of rolls transportation progress are as follows, (a) Heat radiation Heat transfer coefficient due to surface heat radiation: hmid = 4 4   e  Tenv + 273.15   T + 273.15    Tfur − T  100  −   100     (1) Thermal emissivity calculation formula: e = e0 + keT (2) h -surface heat transfer coefficient of rolling plate,  -thermal radiation constant, T -surface temperature of rolling plate, Tenv -ambient temperature, e0 -thermal emissivity constant below 0 °C, ke - slope of thermal emissivity. (b) Convective heat transfer Heat transfer coefficient of bottom surface: hsd =kn  h + hr (3) hsd - heat transfer coefficient on bottom surface of rolling plate, k n -weight, hr -change of heat transfer coefficient caused by roller table contact Other surface heat transfer coefficient: hso =h + hair (4) hso - heat transfer coefficient on other surfaces of rolling plate, hair -air convection heat transfer coefficient. 3.3.2 Rolling progress The rolling process is affected by the contact conduction and heat dissipation mechanism. When the metal goes through the rolling deformation zone, the surface temperature is always in a declining state. In the rolling process, there are three main factors that affect the thermal state of the surface metal: the friction heat generation on the interface, the plastic deformation heat generation of the surface metal, and the contact conduction heat dissipation between the plate and the roll. Due to the chilling effect of the cold roll and the influence of the thermal resistance of the material itself, the temperature in thickness direction gradually increases from the surface to the center. Compared with cold-roll rolling, the use of hot work rolls can effectively reduce the heat loss caused by contact heat conduction, thus effectively improving the temperature uniformity in thickness direction. Due to the short residence time in the rolling deformation zone and the large thermal resistance of the material itself, the temperature of the metal in the slab during rolling mainly depends on the heat generated by plastic deformation, and the influence of the roller temperature change on the slab temperature is difficult to go deeply into the interior. The heat transfer modes of rolling progress are as follows, (a) Roll heat transfer, the factors that affect the heat transfer coefficient of the roll are the heat permeability coefficient and the contact time between the rolled plate and the roll: hrs =kh be tc (5) hrs -heat transfer coefficient of roll, kh -model parameters related to contact time, thickness and heat transmission coefficient, tc -contact time. be = km   m cm  k w   w cw km   m cm + k w   w cw (6) k m - thermal conductivity of rolling plate,  m -density of rolling plate, cm -specific heat of rolling plate， k w - thermal conductivity of work roll,  w -density of work roll, cw -specific heat of work roll. (b) Plasticity work, the influence of plastic deformation on the temperature of the rolled plate is the internal deformation heat and a certain temperature rise directly caused by it. The heat transfer coefficient in the roll gap is: hx =hxg  T + kdT  dT − Tw T[0] − Tw (7) hx - surface heat transfer coefficient of rolled plate, hxg -calculated roll heat transfer coefficient, kdT - effective coefficient of temperature rise during heat conduction, dT -temperature rise caused by the deformation of the rolled plate, T[0] - temperature of the first node in the thickness direction of the rolled plate, Tw -roll temperature. 3.3.3 Coiling progress in furnace Too low temperature during the coiling process will augment the deformation resistance of the magnesium strip, reduce the deformation coordination between the inside and outside of the strip during bending with increasing the power of the coiler, which is not conducive to coiling. In addition, the increase of deformation resistance and springback of magnesium plate will increase the tendency of coil looseness. Equation 8 is the heat transfer coefficient of the middle part of the rolling plate in the coiling furnace, hmid = 4 4   e  T + 273.15   Tfur + 273.15    T − Tfur  100  −   100     (8) Where Tfur - temperature in coiling furnace. Equation 9 is the differential equation of heat conduction inside the coil, T  =   C p   2T  2T  2T Q   2 + 2 + 2 +    x  y  z  C p  (9) Where  is the thermal conductivity; x, y, z are three-dimensional coordinate axes;  , Cp are the density of the steel coil and equal pressure specific heat capacity; T is object transient thermodynamic temperature;  is time; Q is internal heat source, Q is zero when no internal heat source inside the coil. Initial conditions: At the beginning of the process, the temperature in the entire area is known. Boundary conditions: When the coil stays in the hot coil box, the heat loss on the surface mainly includes heat radiation and heat convection. The comprehensive heat transfer coefficient h should be used, as shown below: hr =  (Ts 2 + T f 2 )(Ts + T f )  Ts hd = 1.826  T −T f  s h = f  hr + hd    1 3 (10) (11) (12) Where hr is thermal radiation heat transfer coefficient; hd is thermal convection heat transfer coefficient; Ts is surface temperature of plate; T f is ambient temperature; Emissivity,  =0.7;  is Stefan-Boltzmann constant (5.67×10-8 W/m2·K4); f is radiation heat transfer angle coefficient, there are both radiation and convection on the heat loss of the inner surface of the magnesium coil with the different radiation heat transfer angle coefficients of the inner and outer surfaces. The inner surface is a cylindrical reverse radiating surface, and part of its radiated heat is self-sustained, 0<f<1; f=1 for the outer surface. (b) Radial equivalent thermal conductivity: It is assumed that there is an air gap of constant thickness of about 10% of the thickness of the magnesium plate between two magnesium plates. Baik derived an exponential equation of normal pressure in the radial direction based on the thickness of a unit layer composed of a magnesium layer, an air layer and two oxide layers. This equation has been verified by experiments, and the thermal resistance in the radial direction is expressed by the following formula, kr = where t = ti + ts + 2t0 thickness, , t is unit layer thickness, ti is air layer thickness, ts is plate is oxide layer thickness, t0 (13) t Rs + R0 + 1 / [(1 / Rcd,s )+(1 / Rcd,a )+(1/R rd )] is heat resistance of Mg plate, Rs R0 is oxide and interfacial layer, Rcd,s is thermal resistance conducted through contact points, thermal resistance conducted through air, Rrd Rcd,a is is thermal resistance to radiation through the gap. The above equation considers the influence of surface characteristics, and measures the thickness of the oxide layer. Rs = S s Rcd,s = , S is plate thickness; s is thermal conductivity of Mg plat; p   −0.94 ,  p is surface roughness of Mg plate (μm)； tan  is the average 1.13s tan  slope of the surface shape of the Mg plate; Rcd,a = b (1 −  )g , b is air gap (m),  is the ratio of point contact to apparent area on the interface, g is thermal conductivity of gas; Rrd = 2− 4(1 −  ) Tm 3 , Tm is average temperature of two adjacent Mg plates,  is blackness of Mg plate,  is Stefan-Boltzmann constant. (c) Thermal stress analysis Taking into account the band-shaped protrusion during the coiling process, the compression pressure can be expressed as a function of the hoop stress and the coiling tension in the deformation analysis. P( z ) =  ( z)  t( z) r = E 1− ln ( )  t[1 −  ( rn ( z ) r r z n h ) ] (14) where P( z ) is compression stress,  ( z) is hoop stress, t ( z ) is axial thickness distribution of the plate, t is initial value of the axial thickness of the plate, r is average cylinder radius, rn ( z ) is thin cylinder radius,  is bulge ratio, n is bulge index, h is half width of Mg plate. (d) Considering air heat transfer in the furnace  −n D V  h = K h nD g ( g )  [1 + ( f ) nDL ] g Df L n (15) Where h is convection heat transfer coefficient, V is gas flow rate, Df is gas flow diameter, g is gas thermal conductivity,  g is gas dynamic viscosity,  g is gas density, L is length of gas flow path,  is heat transfer enhancement coefficient caused by swirling flow, K h , nD , n , n , nDL are convection heat transfer coefficient test constant, recommended values are shown in Table 4. Symbol Table 4 Recommendable experiment constant in formula (15) n n nD Kh  Numerical value 1 0.023 0.2 0.8 0.8 nDL 0.7 It can be seen from the above formula that the convection heat transfer coefficient in the furnace is closely related to the flow rate and thermophysical properties of the protective gas. Increasing the gas flow rate in the furnace and reducing the gas dynamic viscosity coefficient can increase the radial heat transfer speed of the magnesium coil, strengthen the convective heat transfer in the furnace, shorten the holding time, and reduce the temperature difference between the inside and outside of the magnesium alloys coil, thereby greatly improving productivity and product quality. The cooling of the magnesium coil went through different stages as the furnace temperature decreases (Fig. 11). The first stage is the high temperature of 350~320°C. At this stage, the temperature of the magnesium sheet is relatively uniform. The dual-temperature zones range is 29.1mm in the rolling direction of the magnesium sheet and 29.3mm in the transverse direction. The second stage is the medium temperature of 210~180°C. At this stage, the cooling range of the edge is enlarged. The range of the two temperature zones in the rolling direction is 32.6mm, and the range of the two temperature zones in the transverse direction is 35.5mm. The last stage is the low temperature of 90~60°C. The cooling range of the edge is further expanded, but the range of the two temperature zones in the rolling direction is reduced to 13.5mm, and the range of the two temperature zones in the transverse direction is expanded to 58.6mm. As the temperature of sheet descended, the temperature difference of sheet width augmented. It could be seen from the cooling curves of the plate from the center to the edge in Fig. 7 and Fig. 9 that the curves became dispersed as the temperature of the plate decreases, indicating that the temperature difference between different parts of the plate becomes larger, which corresponds to the situation in Fig. 11. Fig. 11 Simulation of different cooling stages of plates Table 5 Corresponding relationship between the parameters of the cooling curve and h Sheet specification H6.0 H3.6 Parametric fitting equation R2 A(h) = 20.26 + 169.1exp(−291.3h) 0.9999 B(h) = −0.0003613h −2.228 + 324.4 0.9998 C (h) = 803.9exp(−58.76h) A(h) = 2675 − 2646exp(−0.1775h) 0.9969 B(h) = −0.0108h −1.447 + 327.9 0.9954 C (h) = 575exp(−65.58h) 0.9805 0.8155 Table 5 showed the relationship between each parameter of the formula in the simulation curve of Fig. 9 and the convection heat transfer coefficient h. From Table 5, the fitting accuracy of each parameter of the 6.0mm plate cooling curve is all above 0.9900, while that of the 3.6mm sheet is poor, especially the R2 value of parameter A is only 0.8155, which showed low reliability. The reason is that the heat stored in the thinner plate is less and the convection heat dissipation is faster, resulting in a larger temperature difference between the edge and the center when the temperature is at a low temperature. To resolve the differences between the experimental data and the simulated results of the traditional coiling heat transfer model, a dimensionless correction factor λ was introduced using ANN method and the modified model can be expressed as follows: h =  K h  −n D V  g ( g )  [1 + ( f ) nDL ] nD g Df L n (16) The factor λ was defined as a function of coiling temperature T, plate thickness H and coiling spate v as follows: λ=λ(T, H, v) (17) Therefore, a single-hidden-layer ANN model with the segmental and subregional character of the sheet cooling curve was built to better characterize the temperature change and provide a reference for the temperature change of the sheet coil in the coiling furnace. Fig. 12 showed the temperature discrepancy between various parts of the plate and the center, where the scattered points represented diverse temperature measurement points, and the dotted line is the fitting curve (Te -Edge temperature, Tc-Center temperature, T1/4w-1/4 width temperature) after ANN-modified. It can be seen from Fig. 12 that the temperature difference between the edge Te and the center Tc is the largest when the temperature is lowered to 200s, reaching 15 °C, the temperature difference between the 1/4 width T1/4w and the center Tc is 5 °C, and the average temperature of the plate and the edge temperature difference is 10 °C. It is obvious that the modified model had a better predictability of the temperature difference between different parts during coiling heat transfer process. Experimental value Simulative value Fig. 12 Temperature difference between different parts of the plate and the center Fig. 13 Comparison between fitting surface of modified model and experimental values Fig. 13 shows the description of h under experimental conditions by Eq. (16). It can be seen from the figure that the ANN modified model can well describe the structure-activity relationship between the convection heat transfer coefficient of magnesium plate and temperature, plate thickness and coiling speed during the coiling process. Taking h=0.020 as an example, the segmental and subregional modeling results of sheet cooling based on the ANN are as follows： Table 6 Segmental model of sheet cooling based on the ANN Temperature/Part 1/4Width Fitting formulation 350~320°C T = 33.2377 + 316.7264 exp( − 210~180°C T = 29.2369 + 307.1028exp(− 90~60°C T = 19.1689 + 312.7414 exp( − Center T = 19.3962 + 330.0153exp(− Edge T = 21.4988 + 317.5209 exp(− R2 Reduced Chi-Sqr 0.9999 1.6852 ) 0.9999 2.1020 ) 0.9999 2.7262 0.9999 0.2931 0.9990 5.6338  ) 146.2741  159.1343  171.2994  ) 176.5391  151.7426 ) It could be seen from Table 6 that the cooling model of the edge of the magnesium plate is similar to the cooling model of the 1/4 width at 90~60 °C, which is due to the large heat loss at the edge of the plate and the rapid temperature reduction. 4. Conclusions In order to improve the yield and reduce edge cracks during the rolling process of magnesium coils, the coils are kept warm and supplemented by the coiling furnace, so that the plates can be continuously rolled at a rapid pace, improve production efficiency, and reduce energy consumption. Through the experimental temperature measurement and numerical simulation of veneer and stacked plates with different thicknesses, the following conclusions are drawn: (1) The radial thermal radiation heat transfer rate of the magnesium alloy coil is greater than the thermal conduction heat transfer rate between the coil layers, and the average heating rate of the outer layer of the coil is about twice that of the middle and inner layers. (2) There is a negative correlation between the plate thickness and the heating rate, and the heating rate of the sheet with a small Biot (Bi) value to different temperatures varied greatly. The cooling time of the magnesium plate is proportional to the thickness of the plate, and the cooling rate of the plate with a large Bi value varied greatly from distinct initial temperatures. (3) The heating and cooling models of the uniform thickness plate at different parts of transverse direction are quite discrepant, as well as the various thickness plates to the identical part. (4) A dimensionless correction factor λ was introduced to modify the conventional coil heat transfer models based on the ANN method. The reduced Chi-Spr of the heat transfer model of the coil rolling process is reduced from 6.9195 to 1 nearby. According to the temperature distinction of different parts of the sheet, the precise cooling model is constructed,   T = 19.3962 + 330.0153exp( − ) Center 176.5391    350~320℃ T = 33.2377 + 316.7264 exp( − ) 146.2741    ) 1/ 4Width 210~180℃ T = 29.2369 + 307.1028 exp( − 159.1343    90~60℃ T = 19.1689 + 312.7414 exp( − )  171.2994   T = 21.4988 + 317.5209 exp( − ) Edge 151.7426  Declarations Funding This work is financially supported by the project from the [Key Research and Development Program of Shanxi Province, the National Natural Science Foundation of China, and Taiyuan University of Science and Technology Scientific Research Initial Funding] (Grant numbers 202102050201005, 52105388 and 20222007). Ethics approval and consent to participate The article follows the guidelines of the Committee on Publication Ethics (COPE) and involves no studies on human or animal subjects. Consent for publication This work is approved by all authors for publication. Competing interests The authors have no relevant financial or non-financial interests to disclose. Authors’ Contributions All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Shuping Kong, Fangkun Ning, Lifeng Ma, Weitao Jia and Qichi Le. All authors read and approved the final manuscript References [1] Ning F, Le Q, Yan J, et al. Crimp feasibility of AZ31 magnesium alloy wide plate at warm temperatures responding to asymmetry[J]. Journal of Magnesium and Alloys, 2022, 10(3): 797–810. [2] Liao Q, Lan Q, Jia W, et al. Microstructure and mechanical property inhomogeneities of a modified AZ80 magnesium alloy wide plate under horizontal flat die extrusion[J]. Journal of Materials Engineering and Performance, 2019, 28(4): 1977–1985. [3] Chen S. Magnesium alloy strips production, application status and development prospects[J]. World Nonferrous Metals, 2016(5S): 124–125. [4] Wang Q, Huang S, Wu T. Application analysis of magnesium alloys in weapon components[J]. Ordnance Material Science and Engineering, 2007, 30(2): 69–72. [5] Wang D, Zhao Z. Discuss the application and development of metal materials in automobile lightweight[J]. Auto Expo, 2021(8): 149. [6] Song J. The Application of magnesium alloy in aerospace and national defense[J]. Science Popularization in University, 2018, 12(1): 42–43. [7] Zhang Z. Application of magnesium alloy in defense industry[A]. The 12th annual meeting of Magnesium Branch of China Nonferrous Metals Industry Association[C]. Henan Hebi: 2009. [8] Jia W, Tang Y, Le Q, et al. Air-cooling analysis of AZ31B magnesium alloy plate: Experimental verification, numerical simulation and mathematical modeling[J]. Journal of Alloys and Compounds, 2017, 695: 1838–1853. [9] Jia W, Ma L, Jiao M, et al. Fracture criterion for predicting edge-cracking in hot rolling of twin-roll casted AZ31 Mg alloy[J]. Journal of Materials Research and Technology-JMR&T, 2020, 9(3): 4773–4787. [10] Tian J, Shi Q-X, Meng L-X, et al. Initiation and suppression of crack propagation during magnesium alloy rolling[J]. Materials, 2021, 14(18): 5217. [11] Tian J, Deng J, Shi Q, et al. Effect of temperature field and stress field of different crack behavior on twins and dislocations under Mg alloy rolling[J]. Materials, 2021, 14(19): 5668. [12] Kim M Y. A heat transfer model for the analysis of transient heating of the slab in a direct-fired walking beam type reheating furnace[J]. International Journal of Heat and Mass Transfer, 2007, 50(19): 3740–3748.	17,016
https://openalex.org/W3023481128	OpenAlex	Open Science	CC-By	2,020	An Effective Mental Stress State Detection and Evaluation System Using Minimum Number of Frontal Brain Electrodes	Omneya Attallah	English	Spoken	17,468	33,629	Omneya Attallah Department of Electronics and Communications, College of Engineering and Technology, Arab Academy for Science, Technology and Maritime Transport, Alexandria 1029, Egypt; o.attallah@aast.edu Received: 10 March 2020; Accepted: 1 May 2020; Published: 9 May 2020 Received: 10 March 2020; Accepted: 1 May 2020; Published: 9 May 2020 Abstract: Currently, mental stress is a common social problem aﬀecting people. Stress reduces human functionality during routine work and may lead to severe health defects. Detecting stress is important in education and industry to determine the eﬃciency of teaching, to improve education, and to reduce risks from human errors that might occur due to workers’ stressful situations. Therefore, the early detection of mental stress using machine learning (ML) techniques is essential to prevent illness and health problems, improve quality of education, and improve industrial safety. The human brain is the main target of mental stress. For this reason, an ML system is proposed which investigates electroencephalogram (EEG) signal for thirty-six participants. Extracting useful features is essential for an eﬃcient mental stress detection (MSD) system. Thus, this framework introduces a hybrid feature-set that feeds ﬁve ML classiﬁers to detect stress and non-stress states, and classify stress levels. To produce a reliable, practical, and eﬃcient MSD system with a reduced number of electrodes, the proposed MSD scheme investigates the electrodes placements on diﬀerent sites on the scalp and selects that site which has the higher impact on the accuracy of the system. Principal Component analysis is employed also, to reduce the features extracted from such electrodes to lower model complexity, where the optimal number of principal components is examined using sequential forward procedure. Furthermore, it examines the minimum number of electrodes placed on the site which has greater impact on stress detection and evaluation. To test the eﬀectiveness of the proposed system, the results are compared with other feature extraction methods shown in literature. They are also compared with state-of-the-art techniques recorded for stress detection. The highest accuracies achieved in this study are 99.9%(sd = 0.015) and 99.26% (sd = 0.08) for identifying stress and non-stress states, and distinguishing between stress levels, respectively, using only two frontal brain electrodes for detecting stress and non-stress, and three frontal electrodes for evaluating stress levels respectively. diagnostics diagnostics diagnostics Article Omneya Attallah 1. Introduction Mental stress is the human body’s response to exposed psychosocial or physical situations. It aﬀects people all over the world regardless of their age, gender, or occupation. This is because of the increasing work diﬃculties, rising pressure, and demanding daily activities that people face every day [1]. Currently, mental stress is considered the main cause of several health problems. These problems include heart attacks, strokes, nervousness, depression, post-traumatic stress disorder (PTSD), and immunological disorders. Stress can also inﬂuence brain activity and structure [2]. Therefore, early detection of stress is essential to prevent illness and reduce the chances of clinical brain damage and other health problems. Detection and the evaluation of mental stress is essential as well in ﬁelds such as education and industry. In educational e-learning settings, stress may be a major factor that aﬀects students’ performance in exams. Stress level may increase with unhealthy examination schemes in higher education. In such systems, students can be rated on their performance based on limited hours only. Accordingly, their grades may not represent their real knowledge and intelligence but rather their ability to cope with exam induced stress [3]. Additionally, in an oﬄine educational setting, the frequent evaluation of student’s mental state can be used to deﬁne the speed of teaching and improve the outcome of education [4]. For industrial security, recognizing hazards that occurs due to human mistakes is essential. This is because insecure and careless manners of workers and absence of safety measures are the major reasons for human-caused problems. Such factors include lack of suﬃcient sleep, poor diet, physical deﬁciencies, and fatigue, which can lead a person into a stressful situation. Common methods to measure stress include questionnaires, where the mental eﬀort that applicants put into a task is evaluated [5]. However, such methods can be subjective, i.e., depend on the personal opinion of the applicants, based on psychophysiological [6] or personal measures [7]. Therefore, these methods are not accurate enough due to individuals’ inconsistencies. Moreover, this process becomes challenging when the number of individuals to be evaluated increases in real time. Thus, automated stress detection algorithms that can correctly recognize and assess stress even with a large number of subjects are important in identifying stress factors and facilitating stress management. Omneya Attallah The results show that the proposed system is reliable as the sensitivity is 99.9(0.064), 98.35(0.27), speciﬁcity is 99.94(0.02), 99.6(0.05), precision is 99.94(0.06), 98.9(0.23), and the diagnostics odd ratio (DOR) is ≥100 for detecting stress and non-stress, and evaluating stress levels respectively. This shows that the proposed framework has compelling performance and can be employed for stress detection and evaluation in medical, educational and industrial ﬁelds. Finally, the results veriﬁed the eﬃciency and reliability of the proposed system in predicting stress and non-stress on new patients, as the accuracy achieved 98.48% (sd = 1.12), sensitivity = 97.78% (sd = 1.84), speciﬁcity = 97.75% (sd = 2.05), precision = 99.26% (sd = 0.67), and DOR ≥100 using only two frontal electrodes. Keywords: electroencephalogram (EEG); mental stress detection (MSD); machine learning; mental arithmetic www.mdpi.com/journal/diagnostics Diagnostics 2020, 10, 292; doi:10.3390/diagnostics10050292 Diagnostics 2020, 10, 292 2 of 26 1. Introduction This can assist doctors to understand the signals well, make an accurate diagnosis and provide appropriate treatment [1,8]. Features extracted using feature extraction techniques are fed into a classiﬁcation model to either classify stress, non-stress, or stress levels. Classiﬁcation based on machine learning techniques has the ability to accurately classify stress states. This can assist doctors to understand the signals well, make an accurate diagnosis and provide appropriate treatment [1,8]. Recently, many studies have validated the associations between EEG pattern and emotional stress state [24]. These studies examine stress, non-stress, and stress levels from EEG signals. The authors in [25] proposed a framework that applies the concepts of network physiology and information theory to extract valuable features. A random forest (RF) classiﬁer was constructed to distinguish between stress and non-stress situations. Mahajan, in [18], proposed two feature-sets to classify stress using an artiﬁcial neural network (ANN). In [6], a threshold-based method was introduced to detect stress. The number of peaks of the theta band of an EEG signal was counted. If this count exceeded the threshold, then it was considered a stressful state. The authors of [14] used CSP as a feature extractor. A linear discriminant classiﬁer (LDA) was used to distinguish between stress and non-stress. A support vector machine classiﬁer was employed in [26] to diﬀerentiate between stress levels using power spectral density (PSD) features. PSD features were also used in [19] to discriminate between stress and non-stress conditions using an ANN classiﬁer. In the research articles [20,27], we assessed stress levels using a support vector machine (SVM) classiﬁer. In [28], an SVM classiﬁer was also used to classify the mental stress state of individuals based on the changes in EEG power spectral density, especially in the theta and alpha bands, where the average classiﬁcation accuracy reached 79% and 78%, respectively. The authors in [4] employed a single channel EEG device to examine the use of frontal EEG to determine stress levels. An SVM classiﬁer was employed to evaluate stress levels and the accuracy achieved was in the range of 65%–75%. The authors in [10] employed classical machine learning classiﬁers with a feature selection approach to detect stress levels. In [11], a hybrid feature pool was constructed to recognize stress and non-stress using KNN. Donghoo et al. in [29] proposed a genetic algorithm (GA)-based feature selection algorithm and used and k-nearest neighbor (KNN) classiﬁer to identify stress and non-stress. 1. Introduction These approaches embrace the use of portable devices such as mobiles, remotes, or wearable sensing devices to gather physiological signals, such as electrocardiograms (ECGs), near-infrared spectroscopy (NIRS), near-infrared spectroscopy (NIRS), functional magnetic resonance imaging (fMRI), electro-corticography (ECoG) or electroencephalograms (EEGs) [8,9]. g g g p y p g Recently, brain activity measurement has been veriﬁed as an eﬃcient method in the imaging of emotional stress changes [10]. NIRS and fMRI measure brain activity using blood in the brain. The strength of fMRI is its ability to capture signals in the brain with an outstanding resolution, nevertheless, the measurements are deferred until the state of the brain changes. In contrast, NIRS has the ability to describe only the state of the brain and the signal is eventually captured through blood ﬂow. The EEG and ECoG waves measure the brain signals as well. Regardless of the ability of the ECoG to measure long-bandwidth signals, a surgical procedure is needed to insert electrodes on the skull to detect these signals. EEG is measured non-invasively; it employs a process that requires wearing a helmet. It measures signals from the scalp rather than from the brain itself [3]. Therefore, EEG is preferred over other methods. EEG is commonly used to measure stress [11–14] due to the increasing availability of EEG systems as low-cost wearable devices [15]. Additionally, it has a comparatively greater temporal resolution and it can visualize fast and energetically varying brainwave patterns in complex stress scenarios [9] Therefore, in this study, EEG is used. The main purpose of this paper is to identify the mental state of a person by analyzing the EEG signals. EEG is a noisy signal. Medical signal processing (MSP) techniques play an important role in removing this noise and retaining only the frequency bands containing informative data that describe mental stress. MSP can also extract useful features from EEG signals. Many feature extraction techniques are used for the analysis of EEG signals. Some of these methods include; Hjorth parameters [16,17], power spectral density (PSD) [18,19], common spatial patterns (CSPs) [9,14], statistical-based [20,21], and wavelet-based methods [22,23]. 3 of 26 Diagnostics 2020, 10, 292 Features extracted using feature extraction techniques are fed into a classiﬁcation model to either classify stress, non-stress, or stress levels. Classiﬁcation based on machine learning techniques has the ability to accurately classify stress states. 1. Introduction Fares et al. in [30,31] proposed an MSD to detect stress using an SVM classiﬁer. Most of the previous techniques used a large number of electrodes to construct their MSD systems. The performance of such systems was not suﬃcient enough for real applications. They also lacked eﬃcient feature extraction, selection, or reduction techniques to enhance the performance of a subsequent classiﬁer. p q Other MSD systems were constructed based on deep learning techniques such as Li et al. [24], who used a fused deep learning architecture to extract discriminative spatial–temporal EEG features for detecting emotional stress. Hefron et al. [32] presented a novel convolutional recurrent neural classiﬁer by means of multipath subnetworks for detecting stress. Kuanar et al. [33] built a recurrent neural network algorithm to complete the cognitive analysis of EEG signals. Such methods based on deep learning have several drawbacks. First of all, for eﬃcient performance using deep learning, deep networks usually require large amounts of data for training. Furthermore, deep features extracted from deep networks are commonly not separable, are highly correlated, and have a huge feature dimensional space. Additionally, the current methods do not consider any signiﬁcance to the selection of appropriate features from speciﬁc domains for deeper fundamental analysis. A summary of the recent related techniques from literature that are close to the proposed MSD system are presented in Table 1. 4 of 26 Diagnostics 2020, 10, 292 Table 1. A summary of recent techniques from literature that are comparable to the proposed mental stress detection (MSD) system. Table 1. A summary of recent techniques from literature that are comparable to the proposed mental stress detection (MSD) system. 1. Introduction 5 of 26 Diagnostics 2020, 10, 292 Furthermore, Principal Component Analysis (PCA) feature reduction is applied in a sequential forward search to select the optimal number of principal components and reduce the dimension of feature space, which usually enhances the performance of the subsequent classiﬁer. In addition, in order to make the proposed MSD system more portable and movable and easier to set up, the number of electrodes is further reduced by selecting the minimum number of channels from the site which has the greater impact on the stress detection rates. To prove the eﬃciency of the proposed method, the results are compared with three feature-sets proposed in literature. They are also compared with the state-of-the-art techniques. Furthermore, Principal Component Analysis (PCA) feature reduction is applied in a sequential forward search to select the optimal number of principal components and reduce the dimension of feature space, which usually enhances the performance of the subsequent classiﬁer. In addition, in order to make the proposed MSD system more portable and movable and easier to set up, the number of electrodes is further reduced by selecting the minimum number of channels from the site which has the greater impact on the stress detection rates. To prove the eﬃciency of the proposed method, the results are compared with three feature-sets proposed in literature. They are also compared with the state-of-the-art techniques. 1. Introduction Reference Number of Participants Number of Electrodes Class Label Classiﬁer Validation Method Accuracy [25] 1 14 Stress and non-stress Random Forest k-fold cross validation 97.5% [13] 50 14 Stress and non-stress Threshold-based Hold out 88% [14] 7 31 Stress and non-stress LDA k-fold cross validation 82.6% [19] 5 19 Stress and non-stress ANN Hold out 91.17% [28] 4 22 Stress and non-stress SVM Hold out 78–79% [11] 32 32 Stress and non-stress KNN k-fold cross validation 73.38% [29] 32 32 Stress and non-stress KNN k-fold cross validation 71.76% [31] 25 23 Stress and non-stress SVM k-fold cross validation 83.1%–89.8% [30] 22 7 Stress and non-stress SVM Leave-one out 91.7% [34] 27 5 Stress and non-stress Linear Regression k-fold cross validation 98.76% [35] 50 16 Stress and non-stress Ensemble k-fold cross validation 97.95% [26] 10 14 Stress levels SVM k-fold cross validation 96% [20] 11 14 Stress levels SVM k-fold cross validation 80.32% [27] 42 128 Stress levels SVM k-fold cross validation 94.6% [10] 12 19 Stress levels KNN k-fold cross validation Hold out 91.5% 90.5% [4] 20 1 Stress levels SVM k-fold cross validation 65%–75% [34] 27 5 Stress Levels Linear Regression k-fold cross validation 95.062% [36] 26 9 Stress Levels KNN Leave one out 90.9% [37] 10 4 Stress Levels LDA Leave-one-out 86% [38] 12 7 Stress Levels SVM k-fold cross validation 80%–85% [39] 28 1 Stress Levels SVM k-fold cross validation 78.57% The key aim of this paper is to construct a portable real-time EEG-based mental training neuro-feedback system to identify and evaluate stress levels eﬃciently in real-time. Thus, a new MSD system is proposed to classify stress, non-stress, and stress levels. Relative features are the key factor in a powerful MSD system. Therefore, the proposed technique presents a hybrid feature subset for MSD. Five classiﬁers are used to detect stress situations, then the stress level is assessed and sorted into low and high. In order to generate an eﬃcient MSD system with a lower number of channels, ﬁrst, the proposed MSD scheme explores the placements of electrodes on diﬀerent sites on the skull and selects the location which has the highest inﬂuence on the accuracy of the system. 2.1. Participants Initially, sixty-six subjects (age: µ = 18.6, σ = 0.87) participated in the data collection procedure, and 47 of them were women and 19 were men. According to EEG visual examination by an expert neurophysiologist, thirty of them were omitted from the data collection process because of poor EEG quality (extreme amount of myographic and oculographic artifacts), so the ﬁnal sample size is thirty-six participants [40]. Details of the participants can be found in Table 2. Subjects were qualiﬁed to register in the study if they had no medical signs of cognitive or mental deﬁciency, verbal or non-verbal education incapacities, and had usual or corrected-to-usual visual perception, and normal color sight. Exclusion criteria were the use of medication or alcohol addiction, psychoactive drug, and neurological or illnesses. Table 2. Details of the participants. Name Age (Years) Gender Participant 1 21 Female Participant 2 18 Female Participant 3 19 Female Participant 4 17 Female Participant 5 17 Female Participant 6 16 Female Participant 7 18 Male Participant 8 18 Female Participant 9 26 Male Participant 10 16 Female Participant 11 17 Female Participant 12 18 Female Participant 13 17 Female Participant 14 24 Male Participant 15 17 Female Participant 16 17 Female Participant 17 17 Female Participant 18 17 Female Participant 19 17 Female Participant 20 22 Male Participant 21 17 Female Table 2. Details of the participants. 6 of 26 Diagnostics 2020, 10, 292 Table 2. Cont. Name Age (Years) Gender Participant 22 19 Female Participant 23 20 Female Participant 24 16 Female Participant 25 17 Male Participant 26 17 Male Participant 27 17 Female Participant 28 19 Female Participant 29 19 Female Participant 30 19 Male Participant 31 17 Male Participant 32 19 Female Participant 33 20 Female Participant 34 17 Male Participant 35 18 Female Participant 36 17 Female 2 2 EEG C ll ti P d Name Age (Years) Gender Participant 22 19 Female Participant 23 20 Female Participant 24 16 Female Participant 25 17 Male Participant 26 17 Male Participant 27 17 Female Participant 28 19 Female Participant 29 19 Female Participant 30 19 Male Participant 31 17 Male Participant 32 19 Female Participant 33 20 Female Participant 34 17 Male Participant 35 18 Female Participant 36 17 Female 2.2. EEG Collection Procedure Electrode sites on the scalp are shown in Figure 1. Figure 1. Electrodes placement on the scalp. Figure 1. Electrodes placement on the scalp. Figure 1 Electro 2.3. Proposed Mental Stress Detection System Figure 1 Electro 2.3. Proposed Mental Stress Detection System Figure 1. Electrodes placement on the scalp 2.3. Proposed Mental Stress Detection System In this paper, a new system is proposed to detect stress states and classify stress levels. The proposed system consists of five steps as shown in Figure 2. Initially, the data are preprocessed, and noise and artifact removal constitutes the first step. Subsequently, the data are segmented into frames by a sliding window. Afterwards, valuable features are extracted using time and frequency feature extraction techniques to yield two feature-sets which are combined later to form a hybrid feature-set. Then, a feature reduction step is used to reduce the dimension of the combined feature-set. Finally, five machine classifiers are used to detect stress and evaluate stress levels. The proposed MSD system In this paper, a new system is proposed to detect stress states and classify stress levels. The proposed system consists of ﬁve steps as shown in Figure 2. Initially, the data are preprocessed, and noise and artifact removal constitutes the ﬁrst step. Subsequently, the data are segmented into frames by a sliding window. Afterwards, valuable features are extracted using time and frequency feature extraction techniques to yield two feature-sets which are combined later to form a hybrid feature-set. Then, a feature reduction step is used to reduce the dimension of the combined feature-set. Finally, ﬁve machine classiﬁers are used to detect stress and evaluate stress levels. The proposed MSD system is composed of three experiments. In the ﬁrst experiment, each of the two feature-sets are used individually to construct the MSD system, and then combined to form a hybrid feature-set to examine the inﬂuence of combining time and frequency features. In experiment two, to reduce the number of channels employed to construct the MSD system and enhance the eﬀectiveness of the proposed approach, the electrodes are placed on diﬀerent sites on the skull and the location which has the greatest inﬂuence on the accuracy of the system is chosen. Finally, in experiment three, the PCA feature reduction method is adopted to choose the optimum number of principal components in sequential forward search and reduce the dimension of feature space. 2.2. EEG Collection Procedure 2.2. EEG Collection Procedure Mental arithmetic evaluation is commonly used as a standard stressor [41]. Therefore, in this paper, mental arithmetic tasks are used. The tasks consist of counting during the relaxed state (non-stress) and serial subtraction of two numbers during the stress state. Every serial subtraction trial begins with verbally subtracting 4 numbers from 2. Serial subtraction during 15 min is known as psychosocial stress [42], and it was used in [40]. In this manner, the design of the data collection procedure in [40] required exhaustive cognitive activity from the engaged individuals. This strenuous mental load leads to a variation in the emotional background once the individual involves more eﬀort to solve tasks. Depending on quantity of arithmetic operations per minute, participants were grouped into two classes. The ﬁrst class are called “high stress” and contains participants who performed the arithmetic tasks with diﬃculty and make more eﬀort to perform the arithmetic tasks. The other class is called “low stress” and consists of participants who managed the arithmetic tasks without diﬃculty and no excess eﬀort. Thus in this study, two mental stress states are considered. Subjects were asked to sit on a comfortably reclined armchair in a dark soundproof hall. Before starting the experiment, they were asked to relax throughout the resting state (non-stress state) and informed about the arithmetic task. Subjects were instructed to count accurately and quickly for 3 min without talking or moving in the pace they chose. Afterwards, they performed serial subtraction and the EEG was recorded for 1 min during this state (stress condition). The EEG dataset was acquired using the Neurocom monopolar EEG 23 channel system. Silver/silver chloride electrodes were placed on the scalp to record the data according to the International 10/20 scheme. Electrodes were inserted on diﬀerent locations on the scalp. These sites comprise temporal (T3, T4, T5, T6), frontal (Fp1, Fp2), frontal (F3, F4, Fz, F7, F8), central (C3, C4, Cz), occipital (O1, O2), and parietal (P3, P4, Pz), according to the International 10/20 system. Ear electrodes were used as a reference. Electrode sites on the scalp are shown in Figure 1. 7 of 26 ctrodes Diagnostics 2020, 10, 292 occipital (O1, O2), an d f Diagnostics 2020, 10, 292 occipital (O1, O2), and parietal (P3, P4, Pz), according to the International 10/20 system. Ear elect were used as a reference. 3 1 Data Preprocessing 2.3.1. Data Preprocessing p g An EEG signal is usually noisy due to power line interference, electromyography (EMG), ectrocardiography (ECG), and subject movement, etc. In order to reduce noise and artifacts, a high- ass filter with 0.5 Hz cut-off frequency, low-pass filter with 500 Hz cut-off frequency and a notch lter of 50 Hz were used. The filters used were Butterworth IIR filters of order 4. The distortion of the lters was handled using forward–reverse filtering. In addition, a de-noising method based on multilevel wavelets decomposition was employed [43]. The number of wavelet levels was 5, the mother wavelet was Symlets. The number of vanishing moments was 4. The signals are further moothed using a Savitzky–Golay filter [44] An EEG signal is usually noisy due to power line interference, electromyography (EMG), electrocardiography (ECG), and subject movement, etc. In order to reduce noise and artifacts, a high-pass ﬁlter with 0.5 Hz cut-oﬀfrequency, low-pass ﬁlter with 500 Hz cut-oﬀfrequency and a notch ﬁlter of 50 Hz were used. The ﬁlters used were Butterworth IIR ﬁlters of order 4. The distortion of the ﬁlters was handled using forward–reverse ﬁltering. In addition, a de-noising method based on multilevel wavelets decomposition was employed [43]. The number of wavelet levels was 5, the mother wavelet was Symlets. The number of vanishing moments was 4. The signals are further smoothed using a Savitzky–Golay ﬁlter [44]. 3 2 Segmentation 2.3.2. Segmentation 3.2. Segmentation EEG pre-processed signals are segmented into 4-s segments with a sliding window of 1 s as an ncrement step. The same segmentation procedure as [12] is followed which stated that a window ze of 4 s is suitable and common for classification using a mental arithmetic task. Each of these egments is considered as a single trial. Thus, each trial will have a size of N × T, where N = 19 is the EEG pre-processed signals are segmented into 4-s segments with a sliding window of 1 s as an increment step. The same segmentation procedure as [12] is followed which stated that a window size of 4 s is suitable and common for classiﬁcation using a mental arithmetic task. Each of these segments is considered as a single trial. Thus, each trial will have a size of N × T, where N = 19 is the number of channels and T = 4 corresponds to 4 s of one segmented frame sampled at 500 Hz. All trials will be used later in the feature extraction and classiﬁcation steps. 2.2. EEG Collection Procedure Reducing feature space commonly improves the performance of an MSD system. 8 of 26 pace Diagnostics 2020, 10, 292 8 of 26 quential forward search and reduce the dimension of feature space. Reducing feature space mmonly improves the performance of an MSD system. Figure 2. Block diagram of the proposed system. Figure 2. Block diagram of the proposed system. Diagnostics 2020, 10, 292 equential forward se gure 2. Block diagram of the proposed system. Figure 2. Block diagram of the proposed system. 2.3.3. Feature Extraction To construct a machine learning classiﬁer, useful features are needed to be extracted from the EEG segmented data. For this purpose, for each trial of an EEG signal, two subsets of features are extracted. segmented data. For this purpose, for each trial of an EEG signal, two subsets of features are extracted. Previous studies have demonstrated that to detect emotional activities like stress with higher accuracy, it is preferable to extract features from the frequency domain [45]. Other studies mentioned Previous studies have demonstrated that to detect emotional activities like stress with higher accuracy, it is preferable to extract features from the frequency domain [45]. Other studies mentioned 9 of 26 Diagnostics 2020, 10, 292 that fusing time and frequency features has better impact for the detection rates of stress [46,47]. Therefore, in this paper, we proposed using time and frequency features and studied the impact of fusing these features. The Feature-Set 1 is a frequency feature-set which includes features which were used in [48]. These features are the median frequency (MDF), modiﬁed frequency mean (MFMD) features, and spectral moments (SM). MDF computes the median normalized angular frequency. SM calculates three power spectral moments from each EEG segment corresponding to root squared zero, second and forth order moment. MDMD it is the frequency at which the spectrum is split into two sections with equivalent amplitude. In other words, it determines the median amplitude spectrum in each segment calculated using Fourier transform. Feature-Set 2 consists of the following features; the root mean square (RMS) amplitude of the signal which was used in [49,50] and a sixth order autoregressive (AR) model coeﬃcients which was used in [51,52]. AR uses each sample of EEG segment to describe it as a linear fusion with the preceding samples plus a white noise error term. AR calculates coeﬃcients of the model depending on the order chosen. Such coeﬃcients are considered as features [53]. Feature-Sets 1 and 2 are combined together to form a hybrid subset called Feature-Subset 3. The equations representing the features are shown below. MDF = 1 2 M X j = 1 PSDj (1) (1) MFMD = 1 2 M X k Ak (2) (2) where M is the size of the power spectrum density, and PSDj is the jth line of the power spectrum density; Ak is the EEG amplitude spectrum at frequency index k. 2.3.4. Feature Reduction The hybrid Feature-Set 3 extracted in the previous step is of high dimensional space, and for this reason the feature reduction process is required to lower the dimension of the feature space, reduce the cost of computation of the training process, decrease the complication of the MSD system, improve the eﬃciency of stress detection process, and produce a reliable portable real time MSD system. Principal Component Analysis (PCA) is a common feature reduction technique used to reduce the size of Feature-Set 3 by applying a covariance analysis. PCA shrinks the number of features in Feature-Set 3 to a lower number of principal components [54]. Such principle components convey the variance of the features in Feature-Set 3. The steps of PCA technique used in to reduce Feature-Set 3 are as follows; ﬁrst, we calculate the covariance matrix of Feature-Set 3. Afterwards, we determine the eigenvectors and the eigenvalues of the covariance matrix. Next, we select the number of the principal components using a sequential forward procedure. Finally, a reduced feature-set is produced. 2.3.5. Classiﬁcation The three feature-sets generated in the feature extracted phase are used to construct ﬁve well-known classiﬁcation models. These models are linear discriminate analysis (LDA), k-nearest neighbor (KNN), linear and cubic support vector machine (SVM) classiﬁers, and random forest classiﬁers. The distance metric that is used for the KNN is Euclidian and the number of neighbors (K) is equal to 1. These classiﬁcation models are ﬁrst used to detect stress and non-stress states. Next, they are used to distinguish between two stress levels (low and high stress levels). All models are tested using 5-fold cross-validation. 2.3.3. Feature Extraction where M is the size of the power spectrum density, and PSDj is the jth line of the power spectrum density; Ak is the EEG amplitude spectrum at frequency index k. mo = v u tN−1 X i x2 i (3) m2 = s N−1 P k = 0 k2Pk Pk = 1 N N−1 P k = 0 XkX∗ k (4) m4 = s N−1 P k = 0 k4Pk Pk = 1 N N−1 P k = 0 XkX∗ k (5) (3) (4) (5) where a sampled version of the EEG segment is denoted as xi, with i = 1,2, . . . N, of length N, mo, m2, and m4 are the root squared zero, second and forth order moment. The discrete frequency transform of an EEG segment can be expressed as a function of frequency Xk. Pk is the phase-excluded power spectrum which is equivalent to the result of a multiplication of Xk by its conjugate Xk* divided by N, and k is the frequency index. v where a sampled version of the EEG segment is denoted as xi, with i = 1,2, . . . N, of length N, mo, m2, and m4 are the root squared zero, second and forth order moment. The discrete frequency transform of an EEG segment can be expressed as a function of frequency Xk. Pk is the phase-excluded power spectrum which is equivalent to the result of a multiplication of Xk by its conjugate Xk* divided by N, and k is the frequency index. v RMS = v u t 1 N N X i = 1 x2 i (6) AR = D X d = 1 adxi−d + e (7) (6) (7) 10 of 26 10 of 26 Diagnostics 2020, 10, 292 where ad is AR coeﬃcients, e is white noise or error sequence, and D is the order of AR model, d is the order of the coeﬃcient d = 1,2, . . . ..D. where ad is AR coeﬃcients, e is white noise or error sequence, and D is the order of AR model, d is the order of the coeﬃcient d = 1,2, . . . ..D. 3. Results 3. Results The main objective of this study is to build a portable real-time EEG-based system to detect stress states and distinguish between stress levels. The mental arithmetic test is a popular stress inducer, and so is used in this study. This study presents a new MSD system which consists of four phases, or experiments. Phase one—two feature extraction methods are used to extract valuable features from segmented EEG data. Afterwards, a hybrid feature-set (Feature-Set 3) is formed by fusing feature-set 1 and 2. Phase two—the appropriate electrode site placement is selected, which has higher inﬂuence on stress detection rates. This part examines the electrode placements on diﬀerent sites on the skull and selects the location which has the higher inﬂuence on the accuracy of the system. Phase three—we investigate the optimal number of principal components in the sequential forward search strategy to reduce the dimension of the feature space. Compressing feature space commonly improves the performance of an MSD system. Minimizing the number of EEG channels used in mental stress detection and evaluation would make the system more mobile and easier to set up, and maintain the real-time EEG-based mental stress detection system. Therefore, we come to phase four, where we examine the impact of each frontal electrode and select a minimum number of frontal electrodes in order to construct a portable MSD system. Five classiﬁers were built to identify stress and distinguish between two stress levels (low and high). Figure 3 illustrates the four experiments. The main objective of this study is to build a portable real-time EEG-based system to detect stress states and distinguish between stress levels. The mental arithmetic test is a popular stress inducer, and so is used in this study. This study presents a new MSD system which consists of four phases, or experiments. Phase one—two feature extraction methods are used to extract valuable features from segmented EEG data. Afterwards, a hybrid feature-set (Feature-Set 3) is formed by fusing feature-set 1 and 2. Phase two—the appropriate electrode site placement is selected, which has higher influence on stress detection rates. This part examines the electrode placements on different sites on the skull and selects the location which has the higher influence on the accuracy of the system. Phase three— we investigate the optimal number of principal components in the sequential forward search strategy to reduce the dimension of the feature space. 2.4. Performance Evaluation Several metrics are used to evaluate the performance of the proposed system. These metrics are the classiﬁcation accuracy (CA), sensitivity, speciﬁcity, Goodness Index (G), precision, Matthew correlation coeﬃcient, diagnostic odds ratio (DOR), and receiver operating characteristic (ROC) analysis. CA = TP + TN TN + FP + FN + TP (8) Sensitivity = TP TP + FN (9) Specificity = TN TN + FP (10) GoodnessIndex = q (1 −Sensitivity)2 + (1 −Specificity)2 (11) Precision = TP TP + FP (12) MCC = TP × TN −FP × FN p (TP + FP)(TP + FN)(TN + FP)(TN + FN) (13) DOR = TP × TN FP × FN (14) (8) (9) (10) (13) (14) where TP is the true positive, which is the number of positive class instances that are correctly classiﬁed. TN is the true negative, which is the number of negative class instances that are correctly classiﬁed. FP is the false positive, which is the number of negative class instances that are incorrectly classiﬁed as positive class, and FN is the false negative, which is the number of positive class instances that are incorrectly classiﬁed as negative class. where TP is the true positive, which is the number of positive class instances that are correctly classiﬁed. TN is the true negative, which is the number of negative class instances that are correctly classiﬁed. FP is the false positive, which is the number of negative class instances that are incorrectly classiﬁed as positive class, and FN is the false negative, which is the number of positive class instances that are incorrectly classiﬁed as negative class. Diagnostics 2020, 10, 292 classified as positiv that are incorrectly 11 of 26 instances 11 of 26 instances 3. Results 3. Results Compressing feature space commonly improves the performance of an MSD system. Minimizing the number of EEG channels used in mental stress detection and evaluation would make the system more mobile and easier to set up, and maintain the real-time EEG-based mental stress detection system. Therefore, we come to phase four, where we examine the impact of each frontal electrode and select a minimum number of frontal electrodes in order to construct a portable MSD system. Five classifiers were built to identify stress and distinguish between two stress levels (low and high). Figure 3 illustrates the four experiments. Figure 3. A workflow to explain the three experiments of the proposed MSD system. Figure 3. A workﬂow to explain the three experiments of the proposed MSD system. Figure 3. A workflow to explain the three experiments of the proposed MSD system Figure 3. A workﬂow to explain the three experiments of the proposed MSD system. 3 1 Experiment One Results 3.1. Experiment One Results The classification accuracies for detecting stress and non-stress states for our three subsets of features. 97 97.5 98 98.5 99 99.5 100 LDA Linear SVM Cubic SVM KNN Random Forest feature Set 1 feature Set 2 feature Set 3 Figure 4. The classiﬁcation accuracies for detecting stress and non-stress states for our three subsets of features. Diagnostics. 2020, 10, x FOR PEER REVIEW 12 of 25 Figure 4. The classification accuracies for detecting stress and non-stress states for our three subsets of features. 97 97.5 98 98.5 99 99.5 100 LDA Linear SVM Cubic SVM KNN Random Forest feature Set 1 feature Set 2 feature Set 3 Figure 4. The classiﬁcation accuracies for detecting stress and non-stress states for our three subsets of features. Diagnostics. 2020, 10, x FOR PEER REVIEW 12 of 25 Figure 4. The classification accuracies for detecting stress and non-stress states for our three subsets of features. Figure 4. The classiﬁcation accuracies for detecting stress and non-stress states for our three subsets of features. Diagnostics. 2020, 10, x FOR PEER REVIEW 12 of 25 Table 3. Evaluation metrics (with their 95% confi Accuracy (%) Sensitivity (%) Specificity (% 98.50 (98.49–8.63) 98.89 (98.85–98.93) 98.45 (98.17–98.73) 99.85 (99.8– 99.88) 98.84 (99.81–9.876) 99.86 (99.85–99.88) 99.90 (99.85– 99.93) 99.88 (99.81–99.95) 99.9 (99.88–99.93) 99.86 (99.84–99.89) 99.95 (99.91–99.99) 99.79 (99.75–99.81) t 99.68 (99.64– 99.72) 99.52 (99.42–99.62) 99.76 (99.69–99.83) dence intervals) for detecting stress using Feature-Set 3. Goodness Index Precision (%) MCC (%) 0.0198 (0.0196–0.020) 99.83 (99.69–99.96) 99.7 (99.66–99.76) (0. 0.020 (0.0017–0.002) 99.95 (99.94–99.95) 99.64 (99.57–99.7) (0.5 0.015 (0.0009–0.002) 99.96 (99.94–99.96) 99.7 (99.67–99.74) (6. 0.0022 (0.002–0.0025) 99.99 (99.98–100) 99.64 (99.94–99.98) 0.0054 (0.0044–0.006) 99.8 (99.8–99.8) 99.14 (99.08–99.19) (6.8 Table 3. Evaluation metrics (with their 95% confidence intervals) for detecting stress using Feature-Set 3. Accuracy (%) Sensitivity (%) Specificity (%) Goodness Index Precision (%) MCC (%) 98.50 (98.49–8.63) 98.89 (98.85–98.93) 98.45 (98.17–98.73) 0.0198 (0.0196–0.020) 99.83 (99.69–99.96) 99.7 (99.66–99.76) (0.66 99.85 (99.8– 99.88) 98.84 (99.81–9.876) 99.86 (99.85–99.88) 0.020 (0.0017–0.002) 99.95 (99.94–99.95) 99.64 (99.57–99.7) (0.50 99.90 (99.85– 99.93) 99.88 (99.81–99.95) 99.9 (99.88–99.93) 0.015 (0.0009–0.002) 99.96 (99.94–99.96) 99.7 (99.67–99.74) (6.20 99.86 (99.84–99.89) 99.95 (99.91–99.99) 99.79 (99.75–99.81) 0.0022 (0.002–0.0025) 99.99 (99.98–100) 99.64 (99.94–99.98) t 99.68 (99.64– 99.72) 99.52 (99.42–99.62) 99.76 (99.69–99.83) 0.0054 (0.0044–0.006) 99.8 (99.8–99.8) 99.14 (99.08–99.19) (6.87 Figure 5. Receiver operating characteristic (ROC) curves for detecting stress and non stress; (a) cubic support vector machine (SVM) classifier, (b) k-nearest neighbour (KNN) classifer. Figure 5. 3 1 Experiment One Results 3.1. Experiment One Results 3 1 Experiment One Results 3.1. Experiment One Results 3.1. Experiment One Results Two feature-sets (feature-set 1 and 2) were introduced in our study. Subsequently, a hybrid feature-set (Feature-Set 3) was made by combining these two feature-sets. These feature-sets were ﬁrst used to detect stress and non-stress states. The classiﬁcation accuracy of those three feature-sets, when used for detecting stress and non-stress states, are shown in Figure 4. It is clear from Figure 4 that Feature-Set 3 has higher accuracy than the other two feature-sets except for LDA which has the same performance as feature-set 1. Table 3 shows the evaluation metrics using Feature-Set 3 which achieved a higher accuracy for detecting stress. The sensitivity and speciﬁcity rates are all above 99% except for LDA, which is above 98%, and the sensitivity of linear SVM which is 98.84%. On the Goodness Index, 12 of 26 3 which ve 99% Diagnostics 2020, 10, 292 same performance a achieved a higher ac they are all below 0.02. Figure 5 shows the ROC curve for detecting stress for both a cubic SVM and a KNN classiﬁer. The area under ROC curve (AUC) is one. except for LDA, which is above 98%, and the sensitivity of linear SVM which is 98.84%. On the Goodness Index, they are all below 0.02. Figure 5 shows the ROC curve for detecting stress for both a cubic SVM and a KNN classifier. The area under ROC curve (AUC) is one. they are all below 0.02. Figure 5 shows the ROC curve for detecting stress for both a cubic SVM and a KNN classiﬁer. The area under ROC curve (AUC) is one. except for LDA, which is above 98%, and the sensitivity of linear SVM which is 98.84%. On the Goodness Index, they are all below 0.02. Figure 5 shows the ROC curve for detecting stress for both a cubic SVM and a KNN classifier. The area under ROC curve (AUC) is one. ey are all below 0.02. Figure 5 shows the ROC curve for detecting stress for both a cubic SVM and NN classiﬁer. The area under ROC curve (AUC) is one. oodness Index, they are all below 0.02. Figure 5 shows the ROC curve for detecting stress for bo ubic SVM and a KNN classifier. The area under ROC curve (AUC) is one. Figure 4. Diagnostics 2020, 10, 292 Figure 5 Receive 13 of 26 bic 13 of 26 bic Diagnostics 2020, 10, 292 Figure 5 Receive Afterwards, the three subsets of features are used to classify stress levels. The classiﬁcation accuracy of the three feature-sets used to classify stress levels is shown in Figure 6. Figure 6 shows that Feature-Set 3 yielded the highest accuracies using LDA, cubic and linear SVM, KNN and random forest classiﬁers respectively compared to the other two feature-sets. Table 4 shows the evaluation metric using Feature-Set 3 which achieved the highest accuracy for classiﬁcation of stress levels. Table 3 indicates that the sensitivity ranges between (98%–100%) and the speciﬁcity ranges between (85%–99.4%). Figure 7 shows the ROC curve for detecting stress for both Cubic SVM and KNN classiﬁer. The Goodness Index values are in the range of (0.00634–0.0151). The area under ROC curve (AUC) is one. support vector machine (SVM) classifier, (b) k nearest neighbour (KNN) classifer. Afterwards, the three subsets of features are used to classify stress levels. The classification accuracy of the three feature-sets used to classify stress levels is shown in Figure 6. Figure 6 shows that Feature-Set 3 yielded the highest accuracies using LDA, cubic and linear SVM, KNN and random forest classifiers respectively compared to the other two feature-sets. Table 4 shows the evaluation metric using Feature-Set 3 which achieved the highest accuracy for classification of stress levels. Table 3 indicates that the sensitivity ranges between (98%–100%) and the specificity ranges between (85%– 99.4%). Figure 7 shows the ROC curve for detecting stress for both Cubic SVM and KNN classifier. The Goodness Index values are in the range of (0.00634–0.0151). The area under ROC curve (AUC) is one. Figure 6. The classification accuracies for classifying stress levels using our three subsets of features. 86 88 90 92 94 96 98 100 LDA Linear SVM Cubic SVM KNN Random Forest feature set 1 feature set 2 feature set 3 Figure 6. The classiﬁcation accuracies for classifying stress levels using our three subsets of features. Diagnostics. 2020, 10, x FOR PEER REVIEW 13 of 25 VM 99.7 (99.52–99.79) 99.85 (99.82–99.89) 99.4 (99.05–99.73) 0.00634 (0.0032–0.009) 99.69 (99.61–99.76) 99.49 (99.37–99.62) 2.3 (1.255– N 99 (98.86–99.03) 99.4 (99.25–99.47) 97.7 (97.49–98.10) 0.02296 (0.020–0.0259) 98.27 (97.88–98.66) 97.27 (97.03–97.51) 6.9 (5.426– Forest 98.91 (98.77–99.05) 100 (100–100) 96.02 (95.44–96.59) 0.0398 (0.034–0.0456) 100 (100–100 97.38 (97.19–97.56) > Figure 6. 3 1 Experiment One Results 3.1. Experiment One Results Receiver operating characteristic (ROC) curves for detecting stress and non stress; (a) cubic support vector machine (SVM) classiﬁer, (b) k-nearest neighbour (KNN) classifer. Figure 5. Receiver operating characteristic (ROC) curves for detecting stress and non stress; (a) cubic support vector machine (SVM) classifier, (b) k-nearest neighbour (KNN) classifer. Figure 5. Receiver operating characteristic (ROC) curves for detecting stress and non stress; (a) cubic support vector machine (SVM) classiﬁer, (b) k-nearest neighbour (KNN) classifer. Afterwards, the three subsets of features are used to classify stress levels. The classification Table 3. Evaluation metrics (with their 95% conﬁdence intervals) for detecting stress using Feature-Set 3. Afterwards, the three subsets of features are used to classify stress levels. The classification accuracy of the three feature-sets used to classify stress levels is shown in Figure 6. Figure 6 shows that Feature-Set 3 yielded the highest accuracies using LDA, cubic and linear SVM, KNN and random forest classifiers respectively compared to the other two feature-sets. Table 4 shows the evaluation metric using Feature-Set 3 which achieved the highest accuracy for classification of stress levels. Table 3 indicates that the sensitivity ranges between (98%–100%) and the specificity ranges between (85%– 99.4%). Figure 7 shows the ROC curve for detecting stress for both Cubic SVM and KNN classifier. The Goodness Index values are in the range of (0.00634–0.0151). The area under ROC curve (AUC) is one. 96 98 100 Table 3. Evaluation metrics (with their 95% conﬁdence intervals) for detecting stress using Feature-Set 3. Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Goodness Index Precision (%) MCC (%) DOR LDA 98.50 (98.49–8.63) 98.89 (98.85–98.93) 98.45 (98.17–98.73) 0.0198 (0.0196–0.020) 99.83 (99.69–99.96) 99.7 (99.66–99.76) 1.3 × 106 (0.669–1.952) × 106 Linear SVM 99.85 (99.8–99.88) 98.84 (99.81–9.876) 99.86 (99.85–99.88) 0.020 (0.0017–0.002) 99.95 (99.94–99.95) 99.64 (99.57–99.7) 1.36 × 106 (0.504–3.2306) × 106 Cubic SVM 99.90 (99.85–99.93) 99.88 (99.81–99.95) 99.9 (99.88–99.93) 0.015 (0.0009–0.002) 99.96 (99.94–99.96) 99.7 (99.67–99.74) 8.08 × 105 (6.209–9.946) × 105 KNN 99.86 (99.84–99.89) 99.95 (99.91–99.99) 99.79 (99.75–99.81) 0.0022 (0.002–0.0025) 99.99 (99.98–100) 99.64 (99.94–99.98) >1000 Random Forest 99.68 (99.64–99.72) 99.52 (99.42–99.62) 99.76 (99.69–99.83) 0.0054 (0.0044–0.006) 99.8 (99.8–99.8) 99.14 (99.08–99.19) 7.8 × 104 (6.8783–8.733) × 104 Diagnostics 2020, 10, 292 Figure 5 Receive Evaluation metrics (with their 95% conﬁdence intervals) for evaluating stress levels using Feature-Set 3. Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Goodness Index Precision (%) MCC (%) DOR LDA 98.6 (98.37–98.83) 99 (98.85–99.56) 97 (96.32–97.60) 0.03156 (0.0259–0.037) 97.98 (97.68–98.27) 96.63 (96.25–97) 4589.8 (3451–5728.6) Linear SVM 94.5 (94.08–94.84) 98 (97.83–98.24) 85 (83.93–86.11) 0.151 (0.1401–0.161) 94.16 (93.55–94.77) 85.42 (85.19–85.71) 271.2 (244.74–297.65) Cubic SVM 99.7 (99.52–99.79) 99.85 (99.82–99.89) 99.4 (99.05–99.73) 0.00634 (0.0032–0.009) 99.69 (99.61–99.76) 99.49 (99.37–99.62) 2.39 × 105 (1.255–3.53) × 105 KNN 99 (98.86–99.03) 99.4 (99.25–99.47) 97.7 (97.49–98.10) 0.02296 (0.020–0.0259) 98.27 (97.88–98.66) 97.27 (97.03–97.51) 6.97 × 103 (5.426–8.50) × 103 Random Forest 98.91 (98.77–99.05) 100 (100–100) 96.02 (95.44–96.59) 0.0398 (0.034–0.0456) 100 (100–100 97.38 (97.19–97.56) >1000 In order to show the ability of the proposed feature to distinguish between stress and non-stress scenarios and to separate between stress levels, two ﬁgures are plotted to present two-dimensional scatter plots of two diﬀerent features of the proposed feature-set. Figure 8 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 9 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high and low stress levels cases. These ﬁgures verify the eﬀectiveness of the proposed features to diﬀerentiate between both stress and non-stress, and stress levels. Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classifier, (b) KNN classifier. In order to show the ability of the proposed feature to distinguish between stress and non-stres scenarios and to separate between stress levels, two figures are plotted to present two-dimensiona scatter plots of two different features of the proposed feature-set. Figure 8 shows a two-dimensiona scatter plot of time domain second power spectral moment vs. time domain forth power spectra moment feature for stress and non-stress cases. Figure 9 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature fo high and low stress levels cases. These figures verify the effectiveness of the proposed features to differentiate between both stress and non-stress, and stress levels. In order to show the ability of the proposed feature to distinguish between stress and non-stress scenarios and to separate between stress levels, two ﬁgures are plotted to present two-dimensional scatter plots of two diﬀerent features of the proposed feature-set. Diagnostics 2020, 10, 292 Figure 5 Receive The classification accuracies for classifying stress levels using our three subsets of features. Figure 6. The classiﬁcation accuracies for classifying stress levels using our three subsets of features. (98.86–99.03) (99.25–99.47) (97.49–98.10) (0.020–0.0259) (97.88–98.66) (97.03–97.51) (5.426 orest 98.91 (98 77 99 05) 100 (100 100) 96.02 (95 44 96 59) 0.0398 (0 034 0 0456) 100 (100 100 97.38 (97 19 97 56) Table 4. Evaluation metrics (with their 95% con Feature-Set 3. Accuracy (%) Sensitivity (%) Specificity (%) 98.6 (98.37–98.83) 99 (98.85–99.56) 97 (96.32–97.60) 94.5 (94.08–94.84) 98 (97.83–98.24) 85 (83.93–86.11) Table 4. Evaluation metrics (with their 95% confidence intervals) for evaluating stress levels using Feature-Set 3. Accuracy (%) Sensitivity (%) Specificity (%) Goodness Index Precision (%) MCC (%) 98.6 (98.37–98.83) 99 (98.85–99.56) 97 (96.32–97.60) 0.03156 (0.0259–0.037) 97.98 (97.68–98.27) 96.63 (96.25–97) 94.5 (94.08–94.84) 98 (97.83–98.24) 85 (83.93–86.11) 0.151 (0.1401–0.161) 94.16 (93.55–94.77) 85.42 (85.19–85.71) Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classifier, (b) KNN classifier. Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classiﬁer, (b) KNN classiﬁer. dence intervals) for evaluating stress levels using Goodness Index Precision (%) MCC (%) 0.03156 (0.0259–0.037) 97.98 (97.68–98.27) 96.63 (96.25–97) 0.151 (0.1401–0.161) 94.16 (93.55–94.77) 85.42 (85.19–85.71) Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classifier, (b) KNN classifier. Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classiﬁer, (b) KNN classiﬁer. 14 of 26 Diagnostics 2020, 10, 292 Table 4. Evaluation metrics (with their 95% conﬁdence intervals) for evaluating stress levels using Feature-Set 3. Table 4. Evaluation metrics (with their 95% conﬁdence intervals) for evaluating stress levels using Feature-Set 3. Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Goodness Index Precision (%) MCC (%) DOR LDA 98.6 (98.37–98.83) 99 (98.85–99.56) 97 (96.32–97.60) 0.03156 (0.0259–0.037) 97.98 (97.68–98.27) 96.63 (96.25–97) 4589.8 (3451–5728.6) Linear SVM 94.5 (94.08–94.84) 98 (97.83–98.24) 85 (83.93–86.11) 0.151 (0.1401–0.161) 94.16 (93.55–94.77) 85.42 (85.19–85.71) 271.2 (244.74–297.65) Cubic SVM 99.7 (99.52–99.79) 99.85 (99.82–99.89) 99.4 (99.05–99.73) 0.00634 (0.0032–0.009) 99.69 (99.61–99.76) 99.49 (99.37–99.62) 2.39 × 105 (1.255–3.53) × 105 KNN 99 (98.86–99.03) 99.4 (99.25–99.47) 97.7 (97.49–98.10) 0.02296 (0.020–0.0259) 98.27 (97.88–98.66) 97.27 (97.03–97.51) 6.97 × 103 (5.426–8.50) × 103 Random Forest 98.91 (98.77–99.05) 100 (100–100) 96.02 (95.44–96.59) 0.0398 (0.034–0.0456) 100 (100–100 97.38 (97.19–97.56) >1000 Figure 7. ROC curve for evaluating stress levels; (a) cubic SVM classifier, (b) KNN classifier. In order to show the ability of the proposed feature to distinguish between stress and non stre Table 4. 3.2. Experiment Two Results 3.2. Experiment Two Results As mentioned before, the main aim of the experiment is to explore the electrode locations on different sites on the brain and chooses the site which has the greater impact on the accuracy of the system. This process will also reduce the number of channels employed to construct the MSD system which accordingly makes it more reliable and efficient. Figure 10 shows a bar chart that compares between the performance of several classifiers constructed from Feature-Set 3 extracted from several sites on the skull to detect stress. It is clear from this figure that using only the frontal activation channel, the performance of the MSD system reached 99.98% using the KNN classifier which is higher than all other sites. Table 5 shows the values of accuracy for detecting stress and non-stress from different electrode sites. The frontal activation site has also greater impact on evaluating stress levels as well, shown in Figure 11. Figure 11 shows a bar chart that compares the performance of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to evaluate stress levels. The highest accuracy of 99.78% was achieved using the KNN classifier. Table 6 shows the values of accuracy for evaluating stress levels from different electrode sites As mentioned before, the main aim of the experiment is to explore the electrode locations on diﬀerent sites on the brain and chooses the site which has the greater impact on the accuracy of the system. This process will also reduce the number of channels employed to construct the MSD system which accordingly makes it more reliable and eﬃcient. Figure 10 shows a bar chart that compares between the performance of several classiﬁers constructed from Feature-Set 3 extracted from several sites on the skull to detect stress. It is clear from this ﬁgure that using only the frontal activation channel, the performance of the MSD system reached 99.98% using the KNN classiﬁer which is higher than all other sites. Table 5 shows the values of accuracy for detecting stress and non-stress from diﬀerent electrode sites. The frontal activation site has also greater impact on evaluating stress levels as well, shown in Figure 11. Figure 11 shows a bar chart that compares the performance of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to evaluate stress levels. Diagnostics 2020, 10, 292 Figure 5 Receive Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Figure 9. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Diagnostics 2020, 10, 292 Figure 5 Receive Figure 8 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 9 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high and low stress levels cases. These ﬁgures verify the eﬀectiveness of the proposed features to diﬀerentiate between both stress and non-stress, and stress levels. y p p g scenarios and to separate between stress levels, two figures are plotted to present two-dimensional scatter plots of two different features of the proposed feature-set. Figure 8 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 9 shows a two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high and low stress levels cases. These figures verify the effectiveness of the proposed features to differentiate between both stress and non-stress, and stress levels. Figure 8. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 8. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 8. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. Figure 8. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for stress and non-stress cases. 15 of 26 14 of 25 Diagnostics 2020, 10, 292 Diagnostics. 2020, 10, x F Figure 9. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Figure 9. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Figure 9. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Figure 9. Two-dimensional scatter plot of time domain second power spectral moment vs. time domain forth power spectral moment feature for high- and low-stress level cases. Figure 9. 3.2. Experiment Two Results 3.2. Experiment Two Results The highest accuracy of 99.78% was achieved using the KNN classiﬁer. Table 6 shows the values of accuracy for evaluating stress levels from diﬀerent electrode sites Table 5. Accuracy (95% CI) for detecting stress and non-stress from diﬀerent electrode sites. Classiﬁer Frontal Temporal Central Parietal Occipital LDA 99.3 (99.28–99.32) 98.44 (98.37–98.50) 98.42 (98.36–98.48) 98.56 (98.45–98.67) 98.52 (98.46–98.58) Linear SVM 99.75 (99.67–99.82) 98.31 (98.26–98.38) 98.66 (98.14–99.18) 98.32 (98.18–98.46) 98.24 (98.17–98.30) Cubic SVM 99.91 (99.88–99.93) 99.7 (99.67–99.73) 99.26 (99.19–99.33) 99.46 (99.39–99.53) 99 (98.92–99.12) KNN 99.98 (99.96–100) 99.6 (99.81–99.92) 99.6 (99.56–99.67) 99.4 (99.245–99.55) 99.2 (99.05–99.35) Random Forest 99.67 (99.65–99.69) 99.42 (99.39–99.46) 99.22 (99.19–99.25) 99 (98.97–99.11) 99 (98.96–99.08) Table 5. Accuracy (95% CI) for detecting stress and non-stress from diﬀerent electrode sites. Diagnostics 2020, 10, 292 16 of 26 agnostics. 2020, 10, x FOR PEER REVIEW 15 of Figure 10. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to detect stress and non-stress states. 96 96.5 97 97.5 98 98.5 99 99.5 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Forsest Figure 10. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to detect stress and non-stress states. agnostics. 2020, 10, x FOR PEER REVIEW 16 of g Figure 10. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to detect stress and non-stress states. Table 5. Accuracy (95% CI) for detecting stress and non-stress from different electrode sites. Occipital Parietal Central Temporal Frontal lassifier 98.52 (98.46–98.58) 98.56 (98.45–98.67) 98.42 (98.36–98.48) 98.44 (98.37–98.50) 99.3 (99.28–99.32) DA 98.24 (98.17–98.30) 98.32 (98.18–98.46) 98.66 (98.14–99.18) 98.31 (98.26–98.38) 99.75 (99.67–99.82) near SVM 99 (98.92–99.12) 99.46 (99.39–99.53) 99.26 (99.19–99.33) 99.7 (99.67–99.73) 99.91 (99.88–99.93) ubic SVM 99.2 (99.05–99.35) 99.4 (99.245–99.55) 99.6 (99.56–99.67) 99.6 (99.81–99.92) 99.98 (99.96–100) NN 99 (98.96–99.08) 99 (98.97–99.11) 99.22 (99.19–99.25) 99.42 (99.39–99.46) 99.67 (99.65–99.69) andom Forest 96 96.5 97 97.5 98 98.5 99 99.5 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Forsest Figure 10. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to detect stress and non-stress states. gnostics. 2020, 10, x FOR PEER REVIEW 16 of Figure 11. 3.2. Experiment Two Results 3.2. Experiment Two Results A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to access stress levels. Table 6. Accuracy (95% CI) for evaluating stress levels from different electrode sites. O i i l i l C l T l l Cl ifi 70 75 80 85 90 95 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Figure 11. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to access stress levels. Figure 10. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to detect stress and non-stress states. 96 96.5 97 97.5 98 98.5 99 99.5 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Forsest Figure 10. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to detect stress and non-stress states. agnostics. 2020, 10, x FOR PEER REVIEW 16 of Figure 10. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to detect stress and non-stress states. Figure 10. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to detect stress and non-stress states. nostics 2020, 10, x FOR PEER REVIEW 16 o Table 5. Accuracy (95% CI) for detecting stress and non-stress from different electrode sites. Occipital Parietal Central Temporal Frontal Classifier 98.52 (98.46–98.58) 98.56 (98.45–98.67) 98.42 (98.36–98.48) 98.44 (98.37–98.50) 99.3 (99.28–99.32) DA 98.24 (98.17–98.30) 98.32 (98.18–98.46) 98.66 (98.14–99.18) 98.31 (98.26–98.38) 99.75 (99.67–99.82) inear SVM 99 (98.92–99.12) 99.46 (99.39–99.53) 99.26 (99.19–99.33) 99.7 (99.67–99.73) 99.91 (99.88–99.93) Cubic SVM 99.2 (99.05–99.35) 99.4 (99.245–99.55) 99.6 (99.56–99.67) 99.6 (99.81–99.92) 99.98 (99.96–100) KNN 99 (98.96–99.08) 99 (98.97–99.11) 99.22 (99.19–99.25) 99.42 (99.39–99.46) 99.67 (99.65–99.69) Random Forest Figure 11. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to access stress levels. 3.3. Experiment Three Results The aim of this experiment is to investigate the inﬂuence of reducing the feature space using PCA on the performance of MSD system. The number of principal components are chosen in a sequential forward strategy. Figure 12 shows the selection process for the optimal number of principle components for detecting stress. Figure 12 shows that, using only 58 and 15 principal components, the accuracy of detecting stress reached 100% and 99.8% using cubic SVM and KNN classiﬁers respectively. Diagnostics. 2020, 10, x FOR PEER REVIEW 17 of 25 n the performance of MSD system. The number of principal components are chosen in a sequential orward strategy. Figure 12 shows the selection process for the optimal number of principle components or detecting stress. Figure 12 shows that, using only 58 and 15 principal components, the accuracy of etecting stress reached 100% and 99.8% using cubic SVM and KNN classiﬁers respectively. Diagnostics. 2020, 10, x FOR PEER REVIEW 17 of 2 Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle component for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classifiers respectively. Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle components for valuating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, he accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN lassiﬁers respectively. Figure 12. Selecting the optimal number of principle components for detecting stress. Fi 13 t th l ti d f th ti l b f i i l t Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle components for Figure 12. Selecting the optimal number of principle components for detecting stress Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. 3.2. Experiment Two Results 3.2. Experiment Two Results Table 6 A u a y (95% CI) fo e aluati t e le el f o diffe e t ele t ode ite 70 75 80 85 90 95 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Figure 11. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to access stress levels. Table 5. Accuracy (95% CI) for detecting stress and non-stress from different electrode sites. Occipital Parietal Central Temporal Frontal lassifier 98.52 (98.46–98.58) 98.56 (98.45–98.67) 98.42 (98.36–98.48) 98.44 (98.37–98.50) 99.3 (99.28–99.32) DA 98.24 (98.17–98.30) 98.32 (98.18–98.46) 98.66 (98.14–99.18) 98.31 (98.26–98.38) 99.75 (99.67–99.82) near SVM 99 (98.92–99.12) 99.46 (99.39–99.53) 99.26 (99.19–99.33) 99.7 (99.67–99.73) 99.91 (99.88–99.93) ubic SVM 99.2 (99.05–99.35) 99.4 (99.245–99.55) 99.6 (99.56–99.67) 99.6 (99.81–99.92) 99.98 (99.96–100) NN 99 (98.96–99.08) 99 (98.97–99.11) 99.22 (99.19–99.25) 99.42 (99.39–99.46) 99.67 (99.65–99.69) andom Forest Figure 11. A comparison between classification accuracies of several classifiers constructed using the proposed Feature Set 3 extracted from different sites on the skull to access stress levels 70 75 80 85 90 95 100 Whole Skull Frontal Temporal Central Parietal Occipital LDA Linear SVM Cubic SVM KNN Random Figure 11. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the d F t S t 3 t t d f diﬀ t it th k ll t t l l Figure 11. A comparison between classification accuracies of several classifiers constructed using the proposed Feature-Set 3 extracted from different sites on the skull to access stress levels. Figure 11. A comparison between classiﬁcation accuracies of several classiﬁers constructed using the proposed Feature-Set 3 extracted from diﬀerent sites on the skull to access stress levels. 17 of 26 Diagnostics 2020, 10, 292 Table 6. Accuracy (95% CI) for evaluating stress levels from diﬀerent electrode sites. Classiﬁer Frontal Temporal Central Parietal Occipital LDA 88.74 (88.30–89.18) 90.5 (90.11–90.89) 80.64 (80.39–80.88) 79.92 (79.59–80.05) 75.46 (75.29–75.63) Linear SVM 85.92 (85.34–86.49) 87.5 (87.19–87.76) 76.74 (76.43–77.04) 78 (77.79–78.25) 72.2 (72.2–72.2) Cubic SVM 99 (98.79–99.24) 98.58 (98.31- 98.85) 94.48 (93.96–95.0) 93.32 (92.83–93.80) 90.18 (89.82– 90.53) KNN 99.78 (99.72–99.84) 99 (98.88–99.12) 95.4 (94.96–95.84) 94 (93.81–94.27) 89.84 (89.64–90.03) Random Forest 97.91 (97.73–98.09) 97 (96.73–97.33) 91.96 (91.69–92.22) 91.92 (91.73–92.12) 88.93 (88.64–89.22) 3.3. Experiment Three Results Table 6. Accuracy (95% CI) for evaluating stress levels from diﬀerent electrode sites. 3.2. Experiment Two Results 3.2. Experiment Two Results Linear SVM (85.34–86.49) (87.19–87.76) (76.43–77.04) (77.79–78.25) (72.2–72.2) Cubic SVM 99 (98.79–99.24) 98.58 (98.31- 98.85) 94.48 (93.96–95.0) 93.32 (92.83–93.80) 90.18 (89.82– 90.53) KNN 99.78 (99.72–99.84) 99 (98.88–99.12) 95.4 (94.96–95.84) 94 (93.81–94.27) 89.84 (89.64–90.03) Random Forest 97.91 (97.73–98.09) 97 (96.73–97.33) 91.96 (91.69–92.22) 91.92 (91.73–92.12) 88.93 (88.64–89.22) 3.3. Experiment Three Results The aim of this experiment is to investigate the inﬂuence of reducing the feature space using PCA on the performance of MSD system. The number of principal components are chosen in a sequential forward strategy. Figure 12 shows the selection process for the optimal number of principle components for detecting stress. Figure 12 shows that, using only 58 and 15 principal components, the accuracy of detecting stress reached 100% and 99.8% using cubic SVM and KNN classiﬁers respectively. Diagnostics. 2020, 10, x FOR PEER REVIEW 17 of 25 Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99 4% and 99 7% using cubic SVM and KNN Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN 3.3. Experiment Three Results Figure 13 represents t 3.4. Experiment Four Results for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classifiers respectively. As stated before, one of the aims of this manuscript is to build a portable real-time EEG-based mental training neuro-feedback system to detect and evaluate stress in real time with high accuracy. Minimizing the number of EEG electrodes used in mental stress detection and evaluation would make the system more portable and easier to operate, and maintain the real-time EEG-based mental stress detection system. In this experiment, the impact of each frontal electrode on stress detection and evaluation rates is investigated individually. Afterwards, the inﬂuence of fusing two or three electrodes which have higher detection and evaluation rates when used individually is examined for both stress detection and stress level evaluation. It is clear from Table 7 that Fp1 and Fp2 electrodes have the highest impact on detecting stress and non-stress. Therefore, each one can be used alone to detect stress, and the results show that only one frontal electrode, such as Fp1 or Fp2, is capable of detecting stress and non-stress. However, the impact of fusing Fp1 and Fp2 is examined and shown in Table 8, which shows the impact of using Fp1 and Fp2 on detecting stress and non-stress using ﬁve-fold cross validation. Figure 13. Selecting the optimal number of principle components for evaluating stress levels. 3.4. Experiment Four Results As stated before, one of the aims of this manuscript is to build a portable real-time EEG-base mental training neuro-feedback system to detect and evaluate stress in real time with high accurac Minimizing the number of EEG electrodes used in mental stress detection and evaluation woul make the system more portable and easier to operate, and maintain the real-time EEG-based ment stress detection system In this experiment the impact of each frontal electrode on stress detectio Table 7. Accuracy (%) with (95% CI) for detecting stress and non-stress, and evaluating stress levels using each frontal electrode individually using ﬁve-folds cross validation. 3.3. Experiment Three Results It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classifiers respectively. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classiﬁers respectively. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classifiers respectively. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classiﬁers respectively. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classifiers respectively. Figure 13 represents the selection procedure for the optimal number of principle components for evaluating stress levels. It is clear from Figure 13 that, using only 30 and 54 principal components, the accuracy of classifying stress levels reached 99.4% and 99.7% using cubic SVM and KNN classiﬁers respectively. gnostics 2020, 10, 292 iagnostics. 2020, 10, x FOR PEER REVIEW Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13. Selecting the optimal number of principle components for evaluating stress levels. 18 of 26 17 of 25 Diagnostics 2020, 10, 292 Diagnostics. 2020, 10, x F Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13. Selecting the optimal number of principle components for evaluating stress levels. Figure 12. Selecting the optimal number of principle components for detecting stress. Figure 13. Selecting the optimal number of principle components for evaluating stress levels. Figure 13 represents t 3.4. Experiment Four Results Classiﬁer Fp1 Fp2 F3 F4 FZ F7 F8 Detection of Stress and Non-stress KNN 99.74 (99.70–99.78) 99.54 (99.49–99.59) 99.37 (99.35–99.39) 97.82 (97.77–97.86) 99 (98.98–99.08) 99.29 (99.21–99.37) 98.87 (98.93–99.02) Cubic SVM 99.64 (99.63–99.65) 99.39 (99.35–99.43) 99.18 (99.14–99.22) 97.78 (97.73–97.84) 99.22 (99.20–99.25 99.14 (99.09–99.19) 99 (98.99–99.07) Evaluating Stress Levels KNN 92 (91.71–92.29) 90 (89.82–90.34) 90.74 (90.32–91.16) 85.58 (85.32–85.85) 88.76 (87.96–89.56) 91.36 (90.92–91.79) 92.5 (92.18–92.82) Cubic SVM 88.48 (88.06–88.89) 88.48 (88.21–88.75) 88.62 (88.07–89.17) 87.3 (87.0–87.59) 89.58 (89.29–89.87) 92.26 (91.89–92.63) 90.98 (90.60–91.36) Table 7. Accuracy (%) with (95% CI) for detecting stress and non-stress, and evaluating stress levels using each frontal electrode individually using ﬁve-folds cross validation. 19 of 26 Diagnostics 2020, 10, 292 Table 8. Performance metrics (95% CI) for detecting stress and non- stress with Fp1 and Fp2 electrodes using ﬁve-folds cross validation. Classiﬁer Accuracy Sensitivity Speciﬁcity Precision MCC Goodness Index DOR KNN 99.9 (99.84–99.97) 99.9 (99.65–99.99) 99.94 (99.84–99.98) 99.81 (99.49–99.93) 99.74 (99.71–99.77) 0.006 (0.004–0.007) 1057474 (0.67–1.44) × 106 Cubic SVM 99.75 (99.74–99.77) 99.65 (99.58–99.71) 99.79 (99.77–99.81) 99.37 (99.31–99.43) 99.34 (99.29–99.40) 0.0041 (0.003–0.005) 124250 (1.14–1.35) × 105 Table 8. Performance metrics (95% CI) for detecting stress and non- stress with Fp1 and Fp2 electrodes using ﬁve-folds cross validation. Table 8. Performance metrics (95% CI) for detecting stress and non- stress with Fp1 and Fp2 electrodes using ﬁve-folds cross validation. Classiﬁer Accuracy Sensitivity Speciﬁcity Precision MCC Goodness Index DOR KNN 99.9 (99.84–99.97) 99.9 (99.65–99.99) 99.94 (99.84–99.98) 99.81 (99.49–99.93) 99.74 (99.71–99.77) 0.006 (0.004–0.007) 1057474 (0.67–1.44) × 106 Cubic SVM 99.75 (99.74–99.77) 99.65 (99.58–99.71) 99.79 (99.77–99.81) 99.37 (99.31–99.43) 99.34 (99.29–99.40) 0.0041 (0.003–0.005) 124250 (1.14–1.35) × 105 Table 8 shows that fusing Fp1 and Fp2 improves the performance metric for detecting stress, therefore the two Fp1 and Fp2 electrodes are suﬃcient to construct an eﬃcient and portable MSD system. In order to further validate the performance of the proposed system and its ability to predict if a new person has stress or non-stress, leave-subject-out validation is performed as well and shown in Table 9. The results of Table 9 show the eﬃciency of the proposed system in predicting stress and non-stress on new patients. Table 9. Performance metrics (95% CI) for predicting stress and non-stress with Fp1 and Fp2 electrodes using leave-subjects-out cross validation. Table 9. Performance metrics (95% CI) for predicting stress and non-stress with Fp1 and Fp2 electrodes using leave-subjects-out cross validation. Figure 13 represents t 3.4. Experiment Four Results Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Precision (%) MCC (%) Goodness Index DOR KNN 98.48 (97.47–99.49) 97.78 (97.07–100) 97.75 (95.90–99.59) 99.26 (98.66–99.86) 96.14 (93.69–98.58) 0.032 (0.0041–0.05) >1000 Cubic SVM 98.67 (97.82–99.53) 98.96 (97.67–100) 98.26 (95.68–100) 99.29 (98.44- 100) 96.6 (94.47–98.73) 0.02 (0–0.04) >1000 In the case of evaluating stress levels, Table 7 shows that the highest accuracy achieved for evaluating stress levels using one electrode is 92.5% using the F8 electrode, followed by 92% and 91.36% using Fp1 and F7 respectively. To improve the performance of the proposed system for evaluating stress levels using the minimum number of electrodes, the impact of fusing two or three electrodes is investigated and shown in Table 10. Table 10. Performance metrics (95% CI) of the proposed system constructed using (Fp1 + F7) and (Fp1 + F7 + F8) to evaluate stress levels using ﬁve-folds cross validation. Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Precision (%) MCC (%) Goodness Index DOR Fp1 + F8 KNN 97.52 (94.68–100) 97.59 (97.19–97.97) 99.36 (99.15–99.57) 98.37 (97.9–98.83) 97.32 (96.94–97.69) 0.025 (0.021–0.096) 7708 (0.51–1.03) × 104 Cubic SVM 97.48 (97.37–97.58) 94.96 (94.6–95.27) 98.46 (98.37–98.54) 95.56 (95.75–96.16) 93.73 (93.47–93.97) 0.053 (0.049–0.056) 3611 (1.56–8.78) × 103 Fp1 + F7 + F8 KNN 99.26 (99.17–99.34) 98.35 (98.05–98.64) 99.6 (99.56–99.66) 98.9 (98.65–99.15) 98.15 (97.99–98.32) 0.017 (0.014–0.02) 15935 (1.37–1.81) × 104 Cubic SVM 98.36 (98.25–98.46) 96.65 (96.23–97.07) 99 (98.89–99.15) 97.43 (97.10–97.75) 95.89 (95.62–96.15) 0.035 (0.031–0.04) 2977 (2.6–3.34) × 103 The results of Table 10 show the ability of the proposed system to evaluate level increase using two electrodes. However, the performance of the proposed system is further improved using the three electrodes (Fp1 + F7 + F8). In order to further validate the performance of the proposed system and its ability to predict stress levels for a new person, leave-subject-out validation is performed as well and is Table 10. Performance metrics (95% CI) of the proposed system constructed using (Fp1 + F7) and (Fp1 + F7 + F8) to evaluate stress levels using ﬁve-folds cross validation. The results of Table 10 show the ability of the proposed system to evaluate level increase using two electrodes. However, the performance of the proposed system is further improved using the three electrodes (Fp1 + F7 + F8). Figure 13 represents t 3.4. Experiment Four Results In order to further validate the performance of the proposed system and its ability to predict stress levels for a new person, leave-subject-out validation is performed as well and is The results of Table 10 show the ability of the proposed system to evaluate level increase using two electrodes. However, the performance of the proposed system is further improved using the three electrodes (Fp1 + F7 + F8). In order to further validate the performance of the proposed system and its ability to predict stress levels for a new person, leave-subject-out validation is performed as well and is The results of Table 10 show the ability of the proposed system to evaluate level increase using two electrodes. However, the performance of the proposed system is further improved using the three electrodes (Fp1 + F7 + F8). In order to further validate the performance of the proposed system and its ability to predict stress levels for a new person, leave-subject-out validation is performed as well and is 20 of 26 Diagnostics 2020, 10, 292 shown in Table 11. However, the results of Table 11 show that the proposed system has a lower ability to predict stress levels. Table 11. Performance metrics for predicting stress levels with Fp1 + F8 and Fp1 + F7 + F8 electrodes using leave-subjects-out cross validation. Classiﬁer Accuracy (%) Sensitivity (%) Speciﬁcity (%) Precision (%) MCC (%) Goodness Index DOR Fp1 + F8 KNN 76.85 (67.14–86.58) 67.15 (53.17–81.25) 78.85 (64.54–93.15) 63.83 (49.6–77.56) 48.88 (34.13–63.63) 0.39 (0.2–0.58) 17 (7.669–26.4) Cubic SVM 75.8 (66.59–84.99) 65.26 (50.37–76.14) 80.3 (65.89–94.73) 61.64 (47.85–75.42) 44.87 (30.79–58.95) 0.4 (0.24–0.61) 14.13 (2.675–25.59) Fp1 + F7 + F8 KNN 76.2 (65.77–86.61) 65.66 (48.56–82.77) 80.14 (64.02–96.256 63.48 (47.97–78.99) 47.85 (32.13–63.57) 0.444 (0.295–0.593) 22.28 (0.6879–43.87) Cubic SVM 73.83 (62.82–84.83) 52.04 (34.86–69.22) 77 (58.98–95.13) 60.77 (44.26–77.28) 48 (32.25–63.81) 0.545 (0.424–0.67) 12.39 (3.92–20.86) Table 11. Performance metrics for predicting stress levels with Fp1 + F8 and Fp1 + F7 + F8 electrodes using leave-subjects-out cross validation. 4. Discussion 92 93 94 95 96 97 98 99 100 LDA KNN Linear SVM Cubic SVM Khushaba Feature Set Mahajan Peak Features Mahajan PSD Features Proposed Method Figure 14. The classiﬁcation accuracy of detecting stress and non-stress states using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18] using only Fp1 + Fp2 frontal electrodes. Figure 14. The classification accuracy of detecting stress and non-stress states using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18] using only Fp1 + Fp2 frontal electrodes. Figure 14. The classiﬁcation accuracy of detecting stress and non-stress states using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18] using only Fp1 + Fp2 frontal electrodes. The proposed Feature-Set 3 was also compared with Khushaba et al. and the two feature-sets from Mahajan, but in this case for evaluating stress levels using only Fp1 + F7 + F8 electrodes. Figure 15 shows the classification accuracies of classifying stress levels using our proposed Feature-Set 3 compared to Khushaba and the two feature-sets from Mahajan. It is clear from Figure 15 that the proposed Feature-Set 3 has better performance in evaluating stress levels compared to the other feature-sets from Khushaba et al. and Mahajan. the accuracy of our proposed Feature-Set 3 (82%, 99.26%, 88.1%, and 98.36%,) are higher than the Mahajan PSD feature-set (75.4%, 96.7%, 75.7%, and 94.3%), and Mahajan Peak feature-set (71.7%,72.3%, 63.8%, and 74.6%), using LDA, KNN, linear SVM, and cubic SVM classifiers respectively. The accuracy of the proposed Feature-Set 3 is also greater than Khushaba et al.’s feature-set as well (81.7%, 93.9%, 82.1%, and 95.2%), using LDA, KNN, linear SVM, and cubic SVM classifiers. This suggests that our proposed system, using the hybrid feature-set, is more efficient and has higher ability in identifying stress levels. The proposed Feature-Set 3 was also compared with Khushaba et al. and the two feature-sets from Mahajan, but in this case for evaluating stress levels using only Fp1 + F7 + F8 electrodes. Figure 15 shows the classiﬁcation accuracies of classifying stress levels using our proposed Feature-Set 3 compared to Khushaba and the two feature-sets from Mahajan. It is clear from Figure 15 that the proposed Feature-Set 3 has better performance in evaluating stress levels compared to the other feature-sets from Khushaba et al. and Mahajan. 4. Discussion This study proposes an eﬀective MSD system to detect stress and classify stress levels. For this purpose, the study design consists of three parts. The ﬁrst part compares two feature-sets. The ﬁrst feature-set consists of frequency-based features, whereas the second one consists of AR and RMS features. Here, the inﬂuence of combining time and frequency features is assessed. The analysis showed that combining time and frequency features increases stress detection and stress level classiﬁcation rates. In order to verify the eﬀectiveness of our proposed system, the results are compared with the classiﬁcation accuracy of three feature-sets from literature. These include the feature extraction method proposed by Khushaba et al. [22] and the two approaches presented by Mahajan [18]. The two feature-sets reported by Mahajan include some peak related features and PSD features. Mahajan’s ﬁrst feature-set comprises four peak-related features which are number of negative peaks, the number of positive peaks, the mean of negative peaks, and the mean of positive peaks. The second feature-set of Mahajan’s consists of the mean spectral power estimation in beta (13–30 Hz), alpha (8–13 Hz), theta (4–8 Hz), and delta (0.5–4 Hz) EEG sub-bands respectively. The feature-set of khushaba et al. represents features extracted using wavelet packet decomposition where the frequency components are selected using a mutual information estimation procedure that depends on the generalization of the fuzzy entropy theory. Figure 14 shows the accuracies for detecting stress and non-stress states using our proposed Feature-Set 3 compared to Khushaba and the two feature-sets from Mahajan using only Fp1 + Fp2 frontal electrodes. It is clear from Figure 14 that the accuracy of our proposed Feature-Set 3 (99.94%) is greater than that of Khashaba et al. (98.6%) and the two feature-sets of Mahajan (64.7%, 93.6%) using LDA classiﬁer. Additionally, the accuracies of our proposed Feature-Set 3 (99.93%, 99.7, 99.75%) is greater than that of Khashaba et al. (99.6%, 99.2%, 99.7%), the peak feature-set of Mahajan (96.9%, 97.1%, 92.9%), and the PSD feature-set of Mahajan (99.6%, 98.5%, 99%) using KNN, Linear SVM, cubic SVM classiﬁer respectively. 21 of 26 n r Diagnostics 2020, 10, 292 99.75%) is grea (96.9%, 97.1%, Figure 14. The classification accuracy of detecting stress and non-stress states using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18] using only Fp1 + Fp2 frontal electrodes. 4. Discussion The accuracy of our proposed Feature-Set 3 (82%, 99.26%, 88.1%, and 98.36%,) are higher than the Mahajan PSD feature-set (75.4%, 96.7%, 75.7%, and 94.3%), and Mahajan Peak feature-set (71.7%,72.3%, 63.8%, and 74.6%), using LDA, KNN, linear SVM, and cubic SVM classiﬁers respectively. The accuracy of the proposed Feature-Set 3 is also greater than Khushaba et al.’s feature-set as well (81.7%, 93.9%, 82.1%, and 95.2%), using LDA, KNN, linear SVM, and cubic SVM classiﬁers. This suggests that our proposed system, using the hybrid feature-set, is more eﬃcient and has higher ability in identifying stress levels. Diagnostics. 2020, 10, x FOR PEER REVIEW 21 of 25 Figure 15. The classification accuracies for classifying stress levels using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18]. I th d t f th d t h i th l ti f k ll it d it i fl th 60 65 70 75 80 85 90 95 100 LDA KNN Linear SVM Cubic SVM Khushaba Feature Set Mahajan Peak Features Mahajan PSD Features Proposed Method Figure 15. The classiﬁcation accuracies for classifying stress levels using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18]. Figure 15. The classification accuracies for classifying stress levels using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18]. Figure 15. The classiﬁcation accuracies for classifying stress levels using our proposed Feature-Set 3 compared to Khushaba et al. [22] and the two feature-sets from Mahajan [18]. Diagnostics 2020, 10, 292 22 of 26 In the second part of the proposed technique, the selection of skull site and its inﬂuence on the performance of the MSD system are examined. Based on [4,30,31,55] that showed that EEG signals acquired from the frontal site of the skull are capable of evaluating and detecting the mental stress, this experiment was conducted and proved that frontal brain activation has the most impact on detecting and classifying stress levels. The accuracy of the proposed MSD with frontal brain activation reached 100% for detecting stress and 99.78% for classifying stress levels. The experiment showed that only seven electrodes are enough to achieve a reliable and eﬃcient MSD system. Note that the feature space dimension is 228 after feature extraction. 4. Discussion In the third part of the proposed study, PCA is used to reduce the feature space dimension and to construct the MSD and lower its complexity. Here, we showed that PCA is capable of improving the performance of MSD constructed using a cubic SVM classiﬁer to reach an accuracy of 100% for detecting stress and 99.4% for classifying stress levels with only 58 and 54 principal components. As it is clear from the results, KNN and SVM classiﬁers yielded the highest accuracies for both detections of stress and stress levels. This is because the KNN classiﬁer is simple, straightforward and has high eﬀectiveness, even with noisy datasets [56]. Despite its simplicity, it is able to produce high-accuracy rates in medical applications [57,58]. SVM is a strong classiﬁer, and also has the capability to alter an input vector which is not linearly distinguishable using a hyperplane into a higher-dimensional feature space that is able to linearly discriminate between classes of input data to facilitate the classiﬁcation process. The process is obtained using a kernel function which maps the likeness between the input data and the new higher-dimension feature space. A linear kernel is frequently used when the dataset is just divided by a linear line, which is the case in the feature space used in the case of detecting stress as shown in Figure 8. However, a quadratic kernel is a nonlinear kernel used when the dataset is complex and not linearly separable. Cubic kernels may possibly increase the accuracy. Other beneﬁts of cubic kernels include taking sophisticated mathematical tractability and direct geometric interpretation [51]. The feature space in the case of classifying stress levels was not linearly separable, as shown in Figure 9; therefore, the cubic kernel produced better results than that of the linear kernel in classifying stress levels. EEG signals are commonly known to be non-stationary. This is due the changes in states of neuronal assemblies during brain functioning. It is essential to recognize non-stationaries in EEG signal, because they are illustrative of the underlying actions. This is done by segmenting the EEG signal into smaller stationary segments [59]. Selecting the appropriate window size which segments EEG signal into stationary segments is very important so that the model ﬁts the actual and consistent activity of the brain [60]. There are several ways to check stationarity in literature. Among them, Azami et al. 4. Discussion in [61] suggested that the standard deviation determined for each segment is can be a property that indicates changes in amplitude or/and frequency, as it remains unchanged in stationary intervals, the diﬀerences of the standard deviation in successive windows indicates stationarity. Furthermore, McEwen veriﬁed that short segments (up to 10 s) usually follow the normal distribution while longer segments (up to 60 s) are not Gaussian. McEwen recommended that EEG could be visualized as a procedure consisted from short Gaussian segments. A portion of segments that can be thought as Gaussian reduces from 90% to 20% when the segments’ period rises from 4 to 60 s. In contrast, up to 90% of 4-s-long segments can be believed as stationary while this number decreases to 70%–80% when analyzing 16 s-long EEG segments [2]. Therefore, we used 4-s segment length. It was reported in [62,63] that for the medical system to be reliable, it should achieve a sensitivity greater than or equal to 80%, with a speciﬁcity greater than or equal to 95%, a precision greater than or equal 95%, and a DOR greater than or equal 100. The results in Table 8 show that the proposed system is reliable and can be used to detect stress and non-stress as the sensitivity is 99.9, speciﬁcity is 99.94, precision is 99.81, and DOR is 1057474 using a KNN classiﬁer constructed with only Fp1 and Fp2 frontal electrodes. The results in Table 10 verify that the proposed system is reliable as well as capable of evaluating stress levels, as the sensitivity is 99.6, the speciﬁcity is 98.9, the precision is 98.15, and the DOR is greater than 100 using a KNN classiﬁer constructed with only Fp1 + F7 + F8 frontal electrodes. 23 of 26 Diagnostics 2020, 10, 292 This means that by using only one or two electrodes, the proposed system is capable of detecting stress and non-stress. It is also able to evaluate stress levels using only three frontal electrodes with high performances. The minimum number of electrodes selected to construct the proposed system makes the system more portable and mobile, and easier to set up. Furthermore, to validate the ability of the proposed MSD system to predict if a new person has stress or non-stress, leave-subject-out validation is performed as well. 4. Discussion The results in Table 9 conﬁrm the capability of the proposed system in predicting stress and non-stress using only Fp1 and Fp2 frontal electrodes. They also reveal that the proposed system is reliable and can be used to predict stress and non-stress as the sensitivity is 97.78, the speciﬁcity is 97.75, the precision is 99.26, and the DOR is greater than 100 using a KNN classiﬁer. Finally, the ability of the proposed MSD system to predict the stress level of a new person is tested using leave-subject-out validation. The results in Table 11 shows that the proposed system has lower capability in predicting stress levels. This is due the imbalance occurring between the two classes of stress levels. Therefore, future work will focus on investigating solutions to deal with this imbalance and improve the performance of predicting stress levels. Comparing the results of the proposed algorithm with recent related work in Table 1, it is quite clear that the proposed MSD system outperforms other recent related techniques from literature. More speciﬁcally, the accuracies achieved for the proposed technique are 99.9% (sd = 0.015) and 99.26% (sd = 0.08) for identifying stress and non-stress states, and distinguishing between stress levels, respectively, using only two frontal brain electrodes for detecting stress and non-stress, and three frontal electrodes for evaluating stress levels as shown in Tables 8 and 10. The results prove that the proposed method has a competitive performance compared to the state-of-the-art techniques for both detecting stress and non-stress and classifying stress levels. Conﬂicts of Interest: The authors declare no conﬂict of interest. Funding: This research received no external funding. Conﬂicts of Interest: The authors declare no conﬂict of interest. References 1. Panicker, S.S.; Gayathri, P. A survey of machine learning techniques in physiology based mental stress detection systems. Biocybern. Biomed. Eng. 2019, 39, 444–469. [CrossRef] 2. Al-Shargie, F.; Tang, T.B.; Badruddin, N.; Kiguchi, M. Towards multilevel mental stress assessment using SVM with ECOC: An EEG approach. Med. Biol. Eng. Comput. 2018, 56, 125–136. [CrossRef] [PubMed] 3. Kumari, A.; Jain, J. Examination stress and anxiety: A study of college students. Glob. J. Multidiscip. Stud. 2014, 4, 31–40. 4. So, W.K.; Wong, S.W.; Mak, J.N.; Chan, R.H. An evaluation of mental workload with frontal EEG. 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A Novel Method for Stress Measuring Using EEG Signals. In Proceedings of the Future of Information and Communication Conference, Singapore, Singapore, 4–5 April 2018; pp. 671–684. 5. Conclusions The main aim of the proposed system is to build a portable real-time EEG-based mental training neuro-feedback system to detect and evaluate stress levels in real-time with high accuracy using mental arithmetic tasks. The proposed method introduced a hybrid feature-set (Feature-Set 3) and used ﬁve classiﬁcation models for this purpose. Minimizing the number of EEG channels used in mental stress detection and evaluation would make the system more mobile and easier to set up, and maintain the real-time EEG-based mental stress detection system. This study revealed that the frontal brain activation has a great impact on detecting and evaluating stress levels and is capable of achieving high detection and classiﬁcation rates. Additionally, the study indicated that PCA has the ability to reduce feature space and enhance stress detection rates. Furthermore, the study indicated that only one or two frontal electrodes are capable of detecting stress and non-stress, and three frontal electrodes are able to evaluate stress levels. The results showed that the proposed method based on the hybrid feature-set was capable of both identifying stress and classifying stress levels. Moreover, our method outperformed other feature extraction methods in literature. Furthermore, it has competitive performance compared to the state-of-the-art techniques. Thus, it can be used for stress management, industrial safety, and education. It will enable clinicians to make an accurate diagnosis and provide appropriate treatment, which will consequently reduce chances of clinical brain damage and other health problems. It will also improve the safety standards in industry and will enhance the quality of education. 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https://openalex.org/W2125304008	OpenAlex	Open Science	CC-By	2,013	In vitro antifungal activity of farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida	Jianjun Qiao	English	Spoken	2,879	5,641	© 2013 Qiao et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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. Abstract Background: Protein farnesylation is an important tosttranslational modification in fungi. We evaluated the antifungal activity of two farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida. Methods: Disk diffusion assay and broth microdilution assay were used to determine the antifungal susceptibility of two farnesyltransferase inhibitors (manumycin A and tipifarnib) against clinical isolates of Aspergillus and Candida. antifungal activity of two farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida. Methods: Disk diffusion assay and broth microdilution assay were used to determine the antifungal susceptibility of two farnesyltransferase inhibitors (manumycin A and tipifarnib) against clinical isolates of Aspergillus and Candida. Results: Disk diffusion assay demonstrated both agents had activity against Aspergillus and Candida. The minimal inhibitory concentration (MIC) ranges for manumycin A against Aspergillus and Candida were 200 to 400 μM and 13 to >25 μM, respectively. Unfortunately, the MIC were vastly higher than the concentrations that inhibit the proliferation and viability of mammalian cells. The MICs of tipifarnib against Aspergillus and Candida were >1600 μM. Conclusion: The outcome of present study showed that farnesyltransferase inhibitors have activity against Aspergillus and Candida. This suggests that farnesyltransferase may be used as anifungal target in designing and developing new drugs. Keywords: Aspergillus, Candida, Farnesyltransferase inhibitors, Antifungal susceptibility testing Farnesylation is a posttranslational process that occurs by formation of cysteine thioethers with farnesyl at or near the C-terminus. Farnesyltransferase is a ubiquitous enzyme that catalyzes posttranslational lipidation of the C terminus of more than 60 important signaling proteins in mammalian cells [6]. Several farnesyltransferase inhibitors have been successfully developed as anticancer drugs [6]. Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Open Access In vitro antifungal activity of farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida Jianjun Qiao1†, Peiping Gao2†, Xiaoling Jiang1 and Hong Fang1* Introduction Opportunistic fungal infections, mainly caused by the mold Aspergillus and yeast Candida, are life-threatening in immunocompromised individuals [1,2]. The infections continue to be a serious medical concern and are treated by only a limited number of antifungals: polyenes, azoles, echinocandins, and antimetabolites (flucytosine) [3]. Al- though these drugs are now being used in the prophylaxis and treatment of invasive fungal infection, the number of deaths due to invasive fungal infections remains high. Fur- thermore antifungal resistance and drug toxicity often limit the use of these drugs [3-5]. Therefore, it is needed to discover novel and effective antifungal agents to im- prove the prognosis of invasive fungal infections. In human pathogenic fungi, homologues of farnesyl- transferase were identified [7]. It has been reported that farnesyltransferase inhibitors result in growth reduction of Cryptococcus neoformans [8]. Deletion of genes encoding farnesyltransferase subunit leads to death of C. neoformans and Candida albicans and growth defect in C. glabrata [9-11]. Therefore, this enzyme may be a new target for the development of antifungal agents. In this study, we find that farnesyltransferase inhibitors have in vitro antifungal activity against clinical isolates of Aspergillus and Candida, two important human opportunistic pathogenic fungi. Re- sults of this study may contribute to the development of new antifungals. * Correspondence: fanghongzy@sina.com †Equal contributors 1Department of Dermatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, Zhejiang Province 310003, China Full list of author information is available at the end of the article Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Page 2 of 4 Figure 1 Molecular structures of (a) Manumycin A and (b) tipifarnib. Although both of the chemicals have anti-farnesylation activity, there molecular structures are quite different. Figure 2 Representative effect of manumycin A on the growth of Aspergillus fumigatus. 2 × 105 conidia of A. fumigatus-1 in 200 μl suspension were spread uniformly onto minimal medium agar plate. A 6-mm-diameter paper disk impregnated with 10 μl (20 mM) manumycin A was placed onto the center of the inoculated agar plate. The plate was then incubated at 35°C, and the diameter of the zone of inhibition was measured at 48 h. Figure 2 Representative effect of manumycin A on the growth of Aspergillus fumigatus. 2 × 105 conidia of A. fumigatus-1 in 200 μl suspension were spread uniformly onto minimal medium agar plate. Reagents and antifungals Reagents and antifungals The farnesyltransferase inhibitors (Figure 1) manumycin A (Enzo Life Science International, Inc., USA; molecular weight, 550.7) and tipifarnib (Selleck Chemicals LLC, USA; molecular weight, 489.4) were dissolved in 100% dimethyl sulphoxide. Reference powders of itraconazole, voricona- zole, caspofungin, and amphotericin B were provided by their respective manufacturers. Clinical fungal isolates Clinical fungal isolates used for antifungal susceptibility in this study were obtained from patients in The First Affili- ated Hospital, College of Medicine, Zhejiang University. Introduction A 6-mm-diameter paper disk impregnated with 10 μl (20 mM) manumycin A was placed onto the center of the inoculated agar plate. The plate was then incubated at 35°C, and the diameter of the zone of inhibition was measured at 48 h. Figure 1 Molecular structures of (a) Manumycin A and (b) tipifarnib. Although both of the chemicals have anti-farnesylation activity, there molecular structures are quite different. plates. The plates were then incubated at 35°C, and the di- ameters of the zones of inhibition were measured at 48 h. Broth microdilution assay The broth microdilution assay was done in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines for filamentous fungi (CLSI document M38-A2) and yeasts (CLSI document M27-A3) [12,13]. Stock solu- tions were prepared in RPMI 1640 (Sigma) plates buffered to a pH of 7.0 with 0.165 M 3-(N-morpholino) propanesul- fonic acid (Sigma). The range of final drug concentrations was 16 to 0.03 g/ml for itraconazole, voriconazole, caspo- fungin and amphotericin B, and 1600 to 3 μM for manu- mycin A and tipifarnib. The final inoculum was 0.4 × 104 to 5 × 104 cfu/mL for Aspergillus and 0.5 × 103 to 2.5 × 103 cfu/mL for Candida. The MIC endpoints for manumy- cin A, tipifarnib, itraconazole, voriconazole, and caspofun- gin were designated as the lowest drug concentration that prevented any discernible growth (complete growth inhi- bition) for filamentous fungi or the first well that demon- strated a prominent reduction in growth (a 50% reduction relative to that for the growth control) for yeasts at 48 h of incubation at 35°C. The MIC endpoints for amphotericin B were defined as the lowest drug concentration that com- pletely inhibited fungal growth. Competing interests Competing interests The authors declare that they have no competing interests. Conclusion Our study showed that farnesyltransferase inhibitors have activity against Aspergillus and Candida. This sug- gests that farnesyltransferase may be used as anifungal target in designing and developing new drugs. Results genes should result significant defect of fungal growth [8,9]. We also find that deletion of RAM1 gene in A. fumigatus results in significant growth defect of this fungus (unpublished data). Therefore fungal RAM1 and RAM2 may be a new target for design antifungal drugs. Although the MICs of the current farnesyltrans- ferase inhibitors to Aspergillus and Candida were vastly high, we postulate that designing new farnesyl- transferase inhibitors having high inhibiting effect on fungal farnesyltransferase is promising. Disk diffusion assay results showed that manumycin A produced zones of growth inhibition against the A. fumi- gatus isolates at 48 h of incubation, demonstrating the inherent activities of manumycin A against A. fumigatus (Figure 2). Disk diffusion assay also demonstrated that this drug inhibited growth of A. flavus, A. terreus, and C. albicans. The antifungal activity of manumycin A was further determined by broth microdilution assay, which showed that the MICs of manumycin A to 6 isolates of Aspergillus were 200 to 400 μM and those of 11 isolates of Candida were 13 to 25 μM (Table 1). However disk diffusion assay showed that tipifarnib, another farnesyl- transferase inhibitor produced unclear zones of growth inhibition against both Aspergillus and Candida. Broth microdilution method showed that the MICs of tipifarnib to both Candida and Aspergillus were >1600 μM (Table 1). Both manumycin A and tipifarnib are farnesyltransfe- rase inhibitors, however their chemical structures are quite different (Figure 1). This could explain why the MICs of the two agents against fungi are quite different. Authors’ contributions JQ and PG conducted laboratory analysis and drafted the manuscript. XJ and HF participated in the design of the study. All authors were involved in the interpretation of the data and approved the final manuscript. Disk diffusion assay Disk diffusion assay was performed as described previously [5]. A 200 μl suspension of each tested strain (1 × 106 conidia/ml) was spread uniformly onto minimal medium plates. Blank paper disks 6-mm in diameter were impreg- nated with 10 μl (20 mM) manumycin A or tipifarnib. After drying, they were placed onto the center of inoculated agar C. parapsilosis ATCC 22019 were included in each susceptibility test for quality control and assessment of reproducibility testing [12,13]. Each assay was performed in triplicate on three different days. Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Page 3 of 4 Page 3 of 4 Table 1 Minimal inhibitory concentrations (MICs) of manumycin A, amphotericin B, itraconazole, and voriconazole against Aspergillus and Candida determined by standard broth microdilution method Amphotericin B Itroconazole Voriconazole Caspofungin Manumycin A (μg/ml) (μg/ml) (μg/ml) (μg/ml) (μM) A. fumigatus-1 1 0.5 0.5 0.06 200 A. fumigatus-2 1 0.5 1 0.06 400 A. fumigatus-3 1 0.25 0.5 0.06 200 A. flavus-1 1 0.5 1 0.125 200 A. flavus-2 1 0.5 1 0.125 200 A. terreus-1 2 0.5 0.5 0.06 200 C. albicans-1 0.5 0.25 0.25 1 25 C. albicans-2 1 0.25 0.5 0.5 13 C. albicans-3 1 0.5 0.5 0.5 13 C. albicans-4 0.5 0.25 0.25 1 25 C. tropicalis-1 1 0.125 0.125 0.5 13 C. tropicalis-2 1 0.25 0.125 0.5 25 C. glabrata-1 1 0.5 0.25 1 25 C. glabrata-2 1 0.25 0.125 1 25 C. krusei-1 0.5 1 0.5 0.25 25 C. krusei-2 0.5 1 0.5 0.25 25 C. parapsilosis 0.5 0.25 0.06 1 25 Table 1 Minimal inhibitory concentrations (MICs) of manumycin A, amphotericin B, itraconazole, a against Aspergillus and Candida determined by standard broth microdilution method l inhibitory concentrations (MICs) of manumycin A, amphotericin B, itraconazole, and voriconazole llus and Candida determined by standard broth microdilution method Discussion Recently, it had been reported that farnesyltransferase in- hibitors showed antifungal activity against Cryptococcus [8]. In the present study, we showed that mamumycin A can inhibit growth of Aspergillus and Candida. To our knowledge, this is the first report of the in vitro antifungal activity of manumycin A against Aspergillus and Candida. Acknowledgement The antifungal activity of farnesyltransferase inhibitors suggests that inhibiting or deleting RAM1 and RAM2 This work was supported by The National Natural Science Foundation of China (No. 30900056). Page 4 of 4 Page 4 of 4 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Qiao et al. Annals of Clinical Microbiology and Antimicrobials 2013, 12:37 http://www.ann-clinmicrob.com/content/12/1/37 Received: 30 October 2013 Accepted: 1 December 2013 Published: 5 December 2013 Received: 30 October 2013 Accepted: 1 December 2013 Published: 5 December 2013 Published: 5 December 2013 References 1. Arendrup MC: Epidemiology of invasive candidiasis. Curr Opin Crit Care 2010, 16(5):445–452. 2. Segal BH: Aspergillosis. N Engl J Med 2009, 360(18):1870–1884. 3. Kontoyiannis DP: Invasive mycoses: strategies for effective management. Am J Med 2012, 125(1 Suppl):S25–38. 4. Pfaller MA: Antifungal drug resistance: mechanisms, epidemiology, and consequences for treatment. Am J Med 2012, 125(1 Suppl):S3–13. 5. Qiao J, Kontoyiannis DP, Wan Z, Li R, Liu W: Antifungal activity of statins against Aspergillus species. Med Mycol. 2007, 45(7):589-593. 6. Berndt N, Hamilton AD, Sebti SM: Targeting protein prenylation for cancer therapy. Nat Rev Cancer 2011, 11(11):775–791. 7. Leach MD, Brown AJ: Posttranslational modifications of proteins in the pathobiology of medically relevant fungi. Eukaryot Cell 2012, 11(2):98–108. 8. Hast MA, Nichols CB, Armstrong SM, Kelly SM, Hellinga HW, Alspaugh JA, Beese LS: Structures of Cryptococcus neoformans protein farnesyltransferase reveal strategies for developing inhibitors that target fungal pathogens. J Biol Chem 2011, 286(40):35149–35162. 9. Nakayama H, Ueno K, Uno J, Nagi M, Tanabe K, Aoyama T, Chibana H, Bard M: Growth defects resulting from inhibiting ERG20 and RAM2 in Candida glabrata. FEMS Microbiol Lett 2011, 317(1):27–33. 10. Song JL, White TC: RAM2: an essential gene in the prenylation pathway of Candida albicans. Microbiology 2003, 149(Pt 1):249–259. 11. Vallim MA, Fernandes L, Alspaugh JA: The RAM1 gene encoding a protein- farnesyltransferase beta-subunit homologue is essential in Cryptococcus neoformans. Microbiology 2004, 150(Pt 6):1925–1935. 12. Institute CLS: M27-A3 Reference method for broth dilution antifungal susceptibility testing of yeasts. 3rd edition. Wayne, PA: Clinical Laboratory Standards Institute; 2008. 13. Institute CLS: M38-A2 Reference method for broth dilution antifungal susceptibility testing of filamentous fungi. 2nd edition. Wayne, PA: Clinical Laboratory Standards Institute; 2008. doi:10.1186/1476-0711-12-37 Cite this article as: Qiao et al.: In vitro antifungal activity of farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida. Annals of Clinical Microbiology and Antimicrobials 2013 12:37. References 1. Arendrup MC: Epidemiology of invasive candidiasis. Curr Opin Crit Care 2010, 16(5):445–452. 2. Segal BH: Aspergillosis. N Engl J Med 2009, 360(18):1870–1884. 3. Kontoyiannis DP: Invasive mycoses: strategies for effective management. Am J Med 2012, 125(1 Suppl):S25–38. 4. Pfaller MA: Antifungal drug resistance: mechanisms, epidemiology, and consequences for treatment. Am J Med 2012, 125(1 Suppl):S3–13. 5. Qiao J, Kontoyiannis DP, Wan Z, Li R, Liu W: Antifungal activity of statins against Aspergillus species. Med Mycol. 2007, 45(7):589-593. against Aspergillus species. Med Mycol. 2007, 45(7):589-593. 6. Berndt N, Hamilton AD, Sebti SM: Targeting protein prenylation for cancer therapy. Nat Rev Cancer 2011, 11(11):775–791. 7. Leach MD, Brown AJ: Posttranslational modifications of proteins in the pathobiology of medically relevant fungi. Eukaryot Cell 2012, 11(2):98–108. 8. Hast MA, Nichols CB, Armstrong SM, Kelly SM, Hellinga HW, Alspaugh JA, Beese LS: Structures of Cryptococcus neoformans protein farnesyltransferase reveal strategies for developing inhibitors that target fungal pathogens. J Biol Chem 2011, 286(40):35149–35162. 9. Nakayama H, Ueno K, Uno J, Nagi M, Tanabe K, Aoyama T, Chibana H, Bard M: Growth defects resulting from inhibiting ERG20 and RAM2 in Candida glabrata. FEMS Microbiol Lett 2011, 317(1):27–33. g 10. Song JL, White TC: RAM2: an essential gene in the prenylation pathway of Candida albicans. Microbiology 2003, 149(Pt 1):249–259. g 10. Song JL, White TC: RAM2: an essential gene in the prenylation pathway of Candida albicans. Microbiology 2003, 149(Pt 1):249–259. 11. Vallim MA, Fernandes L, Alspaugh JA: The RAM1 gene encoding a protein- farnesyltransferase beta-subunit homologue is essential in Cryptococcus neoformans. Microbiology 2004, 150(Pt 6):1925–1935. 11. Vallim MA, Fernandes L, Alspaugh JA: The RAM1 gene encoding a protein- farnesyltransferase beta-subunit homologue is essential in Cryptococcus neoformans. Microbiology 2004, 150(Pt 6):1925–1935. 12. Institute CLS: M27-A3 Reference method for broth dilution antifungal susceptibility testing of yeasts. 3rd edition. Wayne, PA: Clinical Laboratory Standards Institute; 2008. 13. Institute CLS: M38-A2 Reference method for broth dilution antifungal susceptibility testing of filamentous fungi. 2nd edition. Wayne, PA: Clinical Laboratory Standards Institute; 2008. doi:10.1186/1476-0711-12-37 Cite this article as: Qiao et al.: In vitro antifungal activity of farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida. Annals of Clinical Microbiology and Antimicrobials 2013 12:37. doi:10.1186/1476-0711-12-37 Cite this article as: Qiao et al.: In vitro antifungal activity of farnesyltransferase inhibitors against clinical isolates of Aspergillus and Candida. Annals of Clinical Microbiology and Antimicrobials 2013 12:37. Author details 1D f 1Department of Dermatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, Zhejiang Province 310003, China. 2Department of pharmacy, Taian City Central Hospital, Taian, Shandong Province, People’s Republic of China. Received: 30 October 2013 Accepted: 1 December 2013 Published: 5 December 2013 References Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission	27,170
https://openalex.org/W3197501191	OpenAlex	Open Science	CC-By	2,021	Characterization of Egyptian Isolates of Canine Distemper and Canine Parvoviruses	M Khoder	English	Spoken	3,692	8,270	Journal of Applied Veterinary Sciences, 6 (4): 01 -06 (October, 2021). ISSN: Online: 2090-3308, Print: 1687-4072 Journal of Applied Veterinary Sciences, 6 (4): 01 -06 (October, 2021). ISSN: Online: 2090-3308, Print: 1687-4072 Characterization of Egyptian Isolates of Canine Distemper and Canine Parvoviruses M.H Khodeir1; Albehwar1 A.M; Omaima A. E. El-Shamandy1; Soliman2 Y. A; El-Dakhly1 A.T. d S h M S b 2 M.H Khodeir1; Albehwar1 A.M; Omaima A. E. El-Shamandy1; Soliman2 Y. A; El-Dakhly1 A.T. and Sahar M. Saber2 1Veterinary Serum and Vaccine Research Institute, Agriculture Research Center, Abasia,Cairo, Egypt. 2Central Laboratory for Evaluation of Veterinary Biologics, Agriculture Research Center, Abasia, Egypt. *Corresponding Author, Khodeir, M.H., E-Mail: profdrkhodeir@gmail.com J. Appl. Vet. Sci., 6(4 ): 01 -06. J. Appl. Vet. Sci., 6(4 ): 01 -06. Canine distemper virus (CDV) has RNA (single-stranded) and belongs to the family paramyxoviridae. The disease is highly contagious via inhalation with morbidity and deaths differ greatly between animal species, with up to 100% doom in non- vaccinated populations. The seriousness and duration of the disease are contingent on the animal's age and the immune state and the degree of virulence of the virus. (Deem et al., 2000 and Andreas et al., 2015). Original Article: g DOI:https://dx.doi.org/10.21608/javs.2 021.84062.1088 This study was aimed to investigate the incidence of CD and CP viruses in Cairo, Egypt. In a private Vet. Clinic in Cairo, five local breed puppies of about 3-5 months of age were supposed to be infected with the virus of canine distemper (CDV) showing fever, runny nose, salivation and coughing. Another 6 German Shepard and 4 Gryphon puppies of about 6-9 months of age were suffered from fever and bloody diarrhea suspected to be infected with canine parvovirus (CPV). All of these puppies had no history of vaccination. Trials of virus segregation in Vero and MDCK cell lines and usage of virus neutralization test (VNT) using specific anti-CD and anti-CP sera revealed that 3 out of 5 nasal swabs were positive to CD virus and 3 out of 10 fecal swabs were positive to CPV. These results were confirmed by Rt-PCR showing positive amplification with CD and CP, respectively. So, we could say that VNT and Rt-PCR are essential assays to identify CD and CP viruses and puppy vaccination should not be neglected to protect them against such diseases. Received :05 July, 2021. Accepted :18 August, 2021. Published in October, 2021. This is an open access article under the term of the Creative Commons Attribution 4.0 (CC- BY) International License . To view a copy of this license, visit: http://creativecommons.org/licenses/by/4.0/ Keywords: Canine Distemper, Canine Parvo, Fluorescent antibody technique, rt-PCR. Keywords: Canine Distemper, Canine Parvo, Fluorescent antibody technique, rt-PCR. 3. Cell cultures: African green monkey kidney and Madin- Darby Canine Kidney Epithelial (MDCK) cell lines propagated with Minimum Essential Medium supplemented with 10% newborn calf serum were used for trials of isolation of CDV and CPV, respectively. 100 microgram of streptomycin and 100 IU of penicillin-G sodium/ml were added to all cell culture media. MATERIIALS AND METHODS The animal may exhibit sensibility to the lighting, incoordination, augmentation sensibility to voluptuous stimuli such as pain or touch, and retrograde motor capabilities. Less commonly, paralysis and blindness may have occurred (Green, 2012). 5. Serologic Detection and Identification of CDV and CPV using of Virus neutralization test (VNT): The obtained CD and CP virus isolates were identified through the application of VNT on the third virus passage of each virus using specific anti-CD and anti-CPV-2 sera according to Yoneda et al., (2008) 1. Diseased dogs: Five puppies of an age range 3-5 months (Local breed) showed fever (39-40 OC); a runny nose, excessive salivation, coughing and/or difficult breathing and loss of appetite suspected to be infected with canine distemper virus. Ten other puppies of an age range 6-9 months (6 German Shepard and 4 Gryphon) were suffered from high fever (41OC), enteritis with vomiting and bloody diarrhea. Two of these puppies were dead within 3 days. Such signs are suspected to be CPV infections. These puppies were found in a private Veterinary Clinic in Cairo without vaccination history. Reverse transcription-polymerase chain reaction (RT-PCR) can be used to reveal viral RNA in respiratory tract secretions, cerebrospinal liquid, stool and urine. This test provides a quantitative measure of the CDV viral load, which is usually much higher during active infection compared to the level detected due to recent vaccination. A stand-alone test is too obtainable for quantitative distemper virus data from collected swabs of respiratory mucosa, preferably deep pharyngeal (University of Wisconsin Madison Shelter Medicine, 2010). 2. Type of Samples: Five nasal swabs were obtained from suspected CD-infected puppies while ten fecal swabs were obtained from suspected CP-infected puppies and subjected to trials of virus isolations. Canine parvovirus (CPV) is a member of the Parvoviridae family and is closely related to feline panleukopenia virus (FPV) and mink enteritis virus (MEV) (Parrish et al., 1982). CPV contains a single- stranded DNA genome surrounded by a protein shell (Carmichael and Joubert 1983 and Kumar et al., 2010). Canine parvovirus infection is a highly contagious and often fatal disease caused by Canine parvovirus 2 (CPV2), the causative factor of myocarditis and acute hemorrhagic enteritis in dogs. It has been well established as an enteric pathogen of dogs throughout the world with a high rate of mortality (100%) (Apple et al., 1979 and Black et al., 1979; Rofaida et al., 2018; Mohamed et al., 2020 and Rabie et al., (2021). The disease is characterized by two prominent clinical forms (i) enteritis with vomiting and diarrhea with heart failure in pups of less than 3 months (Hayes et al., 1979). The virus was termed CPV-2 in order to distinguish it from a closely related parvovirus of canine known as the minute virus of canine (MVC) or CPV-1. INTRODUCTION Infectious diseases threaten dogs like other animal species. Viral infections represent the most dramatic diseases affecting dog populations as Canine Distemper (CD) and Canine Parvo (CP). They usually end with death if the infected dogs did not crescive the correct treatment and management. Canine distemper virus infects the wild and domestic species of dogs, pandas, coyotes, wolves, foxes, skunks, raccoons, and large cats (Ikeda et al., 2001). CD affects several body systems, as well as the respiratory and gastrointestinal tract, the brain and spinal cord, with common signs that include feverish, eye inflammation, discharges from nose and eye, difficult or labored breathing, coughing, nausea, diarrhea, loss of zest, laziness and hardening of nose and footpads. The CD infection can be accompanied by secondary bacteriological infection and can present eventual earnest neurological signs (Ikeda et al., 2001). The distemper signs fluctuate widely from no signs to moderate respiratory signs to severe acute pneumonia with nausea, diarrhea mixed by blood, and death. Other signs may be observed as a runny snoot, diarrhea, vomiting, lack of body fluids, excessive salivation, coughing and/or difficult breathing, loss of zest, and weight loss. Also, there are signs related to the central nervous system including confined involuntary twitching of muscles seizures with excessive salivation and palate movements commonly termed as ―distemper myoclonus‖ or ―chewing-gum fits‖ (Andreas et al., 2015). 1 4. Propagation, detection and isolation of CDV and CPV using cell cultures: Isolation of CDV and CPV was carried out according to Dewey, (2003) and Hirayama et al., (2005) on confluent monolayer Vero and MDCK, respectively through 3 blind successive virus passages. Normal and infected cell cultures were stained with hematoxylin and eosin (H&E) according to Carleton et al., (1967) to describe the induced cytopathic effect (CPE) by the obtained virus isolates. A probable diagnosis of CPV enteritis can be completed based on clinical signs such as gloom, nausea, diarrhea, loss of zest and high fever. The diagnostic tests which were employed earlier include Fluorescent Antibody Test (FAT), (Ramadass and Khadher, 1982), Virus neutralization test, polymerase chain reaction (PCR) (Nandi et al., 2007 and Pereira et al., 2000), real-time qPCR (Decaro et al., 2005), loop-mediated isothermal amplification (LAMP) (Ho- Senog, 2006), nucleic acid sequencing, etc. [Cho et al., 2004 and Nandi et al., 2010). 7.2.Extraction of CPV viral DNA Viral DNA (from parvovirus isolates) was extracted from the purified virus (Sambrook et al., 1989) by Gene JET Genomic DNA Purification Kit (Thermo Scientific Cat. # K0721) as stated by the manufacturer's instructions and stored at -20°C till used. Photo (1-B): CDV infected Vero cell culture (H&E, 100X) showing represented by giant cell formation. Table 2: Molecular Detection and Identification of CDV using RT-PCR. Table 1: The used primers for detection of CD and CP viruses Name Seq 5’ → 3’ Tyarged gene ref CDVF4 GTCGGTAATCGAGGATTCGAGAG phosphoprotein (P) gene of canine distemper virus 1 and 2 CDVR3 GCCGAAAGAATATCCCCAGTTAG CDV probe FAM-ATCTTCGCCAGAATCCTCAGTGCT-BHQ1 Parvo-F ACACCTgAgAgATTTACATATATAgCACA VP2 of parvo 3 Parvo-A ATTAgTATAgTTAATTCCTgTTTTACCTCC FAM-AATACATXITATCATCTgTTACAggAAgg- BHQ1 7. Molecular Detection and Identification of CDV and CPV using of RT-PCR: 7.1. Extraction of CDV viral RNA: 6.Primers used: The used primers for detection of CD and CP viruses in the present study are tabulated in table (1) according to Scagliarini et al., (2007); Tomaszewicz Brown et al., (2020) and Hoang et al., (2019) The current work aimed to isolate and characterize CD and CP viruses recently affecting dogs in Cairo Province, Egypt. 2 Khodeir, M.H. et al……. Table 1: The used primers for detection of CD and CP viruses Name Seq 5’ → 3’ Tyarged gene ref CDVF4 GTCGGTAATCGAGGATTCGAGAG phosphoprotein (P) gene of canine distemper virus 1 and 2 CDVR3 GCCGAAAGAATATCCCCAGTTAG CDV probe FAM-ATCTTCGCCAGAATCCTCAGTGCT-BHQ1 Parvo-F ACACCTgAgAgATTTACATATATAgCACA VP2 of parvo 3 Parvo-A ATTAgTATAgTTAATTCCTgTTTTACCTCC FAM-AATACATXITATCATCTgTTACAggAAgg- BHQ1 Table 1: The used primers for detection of CD and CP viruses 7. Molecular Detection and Identification of CDV and CPV using of RT-PCR: 7.1. Extraction of CDV viral RNA: Photo (1-B): CDV infected Vero cell culture (H&E, 100X) showing represented by giant cell formation. Viral RNA (from Canine distemper isolates) was extracted from the purified virus (Sambrook et al., 1989), by QIA amp Viral RNA Mini Kit (Qiagen Germany, cat #52904), according to the manufacture instructions. 8. QRT-PCR: Well Well Name Threshold (dR) Ct (dR) RSq (dR) Slope (dR) A3 CD sample 1 270.332 32.89 0.999 -2.236 A4 CD sample 2 270.332 34.98 0.999 -2.236 A5 CD sample 3 270.332 37.36 0.999 -2.236 The amplification response was completed in MX3005P real-time PCR (Agilent, USA). The reaction mix per 25µl was 10 ng of the extracted RNA, 12.5µl of 2XBrillient II one step qRT-PCR master mix (Agilent cat # 600809), 50pmol of each primer and 100 pmol of each probe. The cycling condition was as adjusted at 50°C/30min for reverse transcription and initial denaturation phase at 95 °C for 10 minutes, 40 cycles of 95°C for 15 seconds, 50 °C for 20 seconds, and 70 °C for 20 seconds, the fluorescence emission for FAM was adjusted to be collected at the terminus of each extension stage. j of each extension stage. RESULTS Photo (1-A): Normal Vero cell culture (H&E, 100xs) Fig. 1: The amplification plot of real-time RT-PCR of the CD tested sample. Fig. 1: The amplification plot of real-time RT-PCR of the CD tested sample. RESULTS Photo (1-A): Normal Vero cell culture (H&E, 100xs) DISCUSSION Well Well Name Well Comment Threshold (dRn) Ct (dRn) A4 Parvo sample 5 H4 0.1898 33.38 A6 Parvo sample 4 H6 0.1898 32.8 A7 Parvo sample 7 H7 0.1898 35.03 A8 Parvo sample 6 K2 0.1898 No Ct Well Well Name Well Comment Threshold (dRn) Ct (dRn) A4 Parvo sample 5 H4 0.1898 33.38 A6 Parvo sample 4 H6 0.1898 32.8 A7 Parvo sample 7 H7 0.1898 35.03 A8 Parvo sample 6 K2 0.1898 No Ct Fig. 2: The amplification plot of real-time PCR of the Parvo tested samples. Well Well Name Well Comment Threshold (dRn) Ct (dRn) A4 Parvo sample 5 H4 0.1898 33.38 A6 Parvo sample 4 H6 0.1898 32.8 A7 Parvo sample 7 H7 0.1898 35.03 A8 Parvo sample 6 K2 0.1898 No Ct RESULTS Fig. 1: The amplification plot of real-time RT-PCR of the CD tested sample. Fig. 1: The amplification plot of real-time RT-PCR of the CD tested sample. Photo (1-A): Normal Vero cell culture (H&E, 100xs) 3 Characterization of Egyptian Isolates ……… Characterization of Egyptian Isolates ……… CONCLUSION Depending on the data of the present work, it could be concluded that neglection of puppy vaccination against infectious diseases; especially those of dramatic forms as CD and CP; resulted in puppy infection, which could spread the viruses in the environment and disease outbreaks. So, it could be stated that virus isolation and identification using specific and accurate assays as VN and RT-PCR are essential steps to reach correct disease diagnosis aiming to control infectious diseases. In addition, identifying the potential risk factors of CD and CPV infections may help apply preventive measures. Fig. 2: The amplification plot of real-time PCR of the Parvo tested samples. Fig. 2: The amplification plot of real-time PCR of the Parvo tested samples. DISCUSSION Photo (2-A): Normal MDCK (H&E, 100xs) The recorded clinical signs in puppies suffering from fever, vomiting and bloody diarrhea directed the attention toward canine parvovirus infection where such symptoms were stated by Black et al., 1979; Woods et al., 1980; Rofaida et al., 2018; Mohamed et al., 2020 and Rabie et al., 2021). Infection of MDCK (Madin-Darby Canine Kidney) cell line with the prepared fecal samples from suspected CP infected dogs; through three successive passages revealed that 3 samples out of 10 showed cell rounding and aggregation followed by cell lysis and detachment of the cell sheet (photo 2-B). In this respect, it was concluded that cell lines like MDCK (Madin-Darby Canine Kidney) support replication of CPV and virus could be isolated from the cases of CPV and the cell culture adapted virus will enable the biochemical and molecular description of the CPV isolates (Appel et al., 1979; Black et al., 1979) describing the same CPE findings. Photo (2-A): Normal MDCK (H&E, 100xs) Photo (2-B): CPV infection in MDCK (4days post cell infection; H&E, 100xs). Identification of the obtained 3 positive samples in MDCK CPV was carried out by applying VNT using specific anti-CPV-2 serum as a specific, accurate test as concluded by Nandi et al., 2010; Rofaida et al., 2018 and Mohamed et al., 2020). Photo (2-B): CPV infection in MDCK (4days post cell infection; H&E, 100xs). Regarding the results of Rt-PCR, the tested samples 4,5 and 7 gave a positive amplification with Ct while sample 6 was negative as shown in table (3) and amplification lot as in Fig (2). Real-time PCR (RT- PCR) employing has been used for the detection of CPV-2 DNA and it was applied to obtain rapid and unambiguous identification of the viral type (Decaro et al., 2005 and 2006; Rofaida et al., 2018 and Rabie et al., 2021). Table 3: Molecular Detection and Identification of CPV using of RT-PCR. Fig. 2: The amplification plot of real-time PCR of the Parvo tested samples. REFERENCES ANDREAS, BEINEKE; BAUMGÄRTNER, WOLFGANG; WOHLSEIN, PETER, 2015. Cross- species transmission of canine distemper virus—an update." One Health. 1: 49–59. HOANG, M., WU, H.Y., LIEN, Y.X., CHIOU, M.T., LIN, C.N., 2019. A SimpleProbe® real-time PCR assay for differentiating the canine parvovirus type 2 genotype. J Clin Lab Anal. 2019;33: e22654 https://doi.org/10.1002/jcla.22654 APPEL, MJG., COOPER, BJ., GREISEN, H., and CARMICHAEL LE., 1978. Status report: canine viral enteritis. J Am Vet Med Assoc; 173:1516–1518. IKEDA, YASUHIRO; NAKAMURA, KAZUYA; MIYAZAWA, TAKAYUKI; CHEN, MING-CHU; KUO, TZONG-FU; LIN, JAMES A; MIKAMI, TAKESHI; KAI, CHIEKO and TAKAHASHI, EIJI., 2001. "Seroprevalence of Canine Distemper Virus in Cats." Clin Vaccine Immunol. 8 (3):641–644. APPEL, MJG., SCOTT, FW., and CARMICHAEL, LE., 1979. 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Declaration of Conflicting Interests Fig. 2: The amplification plot of real-time PCR of the Parvo tested samples. The authors revealed that there was no potential conflicts of interest. 4 Khodeir, M.H. et al……. parvovirus isolate from stool of a puppy, J. Vet. Med. Scib. 67: 139-143 Characterization of Egyptian Isolates ……… characterization of canine parvovirus in dogs; EJPMR, 5 (4): 525-535 characterization of canine parvovirus in dogs; EJPMR, 5 (4): 525-535 How to cite this article: Khodeir, M.H., Albehwar, A.M., Omaima, A. E. El- Shamandy., Soliman, Y. A., El-Dakhly, A.T. and Sahar M. Saber, 2021. Characterization of Egyptian Isolates of Canine Distemper and Canine Parvoviruses. Journal of Applied Veterinary Sciences, 6 (4): 01 – 06. DOI:https://dx.doi.org/10.21608/javs.2021.84062.1088 How to cite this article: Khodeir, M.H., Albehwar, A.M., Omaima, A. E. El- Shamandy., Soliman, Y. A., El-Dakhly, A.T. and Sahar M. Saber, 2021. 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Antigenic relationships between canine parvovirus type 2, feline panleukopenia virus and mink enteritis virus using conventional antisera and monoclonal antibodies. Archives of Virology 72, 267– 278 GABRIELLA ELIA, NICOLA DECARO, VITO MARTELLA, and FRANCESCO CIRONE, 2006. Detection of canine distemper virus in dogs by real- time RT-PCR; Journal of Virological Methods 136 (1- 2):171-6. GREENE, CE., VANDEVELDE, M., 2012. "Chapter 3: Canine distemper". In Greene, Craig E. (ed.). Infectious diseases of the dog and cat (4th ed.). St. Louis, Mo.: Elsevier/Saunders. pp. 25–42 PEREIRA, CA., MONEZI, TA., MEHNERT, DU., D’ANGELO, M., and DURIGON, EL., 2000. Molecular characterization of canine parvovirus in Brazil by PCR. Vet Microbiol., 75:127–133. HAYES, MA., RUSSELL, RG., MUELLER, RW., and LEWIS, RJ., 1979. Myocarditis in young dogs associated with a parvovirus-like agent. Can Vet J; 20:126–132. RABIE ETMAN; MAHMOUD, S., SAFWAT, and MOHAMED, H. KHODEIR, 2021. 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