bluuebunny/crossref_metadata_2025 huggingface dataset:
Enzimas detoxificantes asociadas con la resistencia del pulgón amarillo (Melanaphis sacchari) a insecticidas en el centro del estado de Guanajuato, México
Authors: García-Burgos Enrique , Landeros-Flores Jerónimo , Cerna-Chávez Ernesto , Ochoa-Fuentes Yisa María , Aguirre-Uribe Luis Alberto , Guevara-Acevedo Luis Patricio , Rodríguez-Pérez Gilberto
Published: February 2021

El pulgón amarillo (Melanaphis sacchari) considerado como la plaga más devastadora en el cultivo del sorgo, se ha extendido por casi todo el territorio mexicano provocando bajos rendimientos, especialmente en los lugares con mayor producción agrícola. En el estado de Guanajuato ha ocasionado severos daños y debido a la importancia del cultivo y al perjuicio generado, se ha convertido en un grave problema. Su control se basa principalmente en el método químico y debido a esta situación para controlarlo, los insecticidas pueden conferirle desarrollo de resistencia a los diferentes ingredientes activos utilizados para su control. Por tal motivo, el objetivo fue cuantificar cinco enzimas detoxificantes en 13 municipios del estado de Guanajuato. Las enzimas con mayor presencia fueron α-esterasas,β-esterasas, además de oxidasas.
Conceptualización, manejo y monitoreo de áreas naturales protegidas en México: Caso Reserva Estatal Sierra Monte Negro, Morelos
Authors: Vázquez-Márquez Gustavo Emmanuel , Ramírez-García Adán Guillermo , Palacios-Rangel María Isabel , Monterroso-Rivas Alejandro Ismael
Published: February 2021

Las áreas naturales protegidas (ANP) gestionan los recursos naturales con el fin de preservarlos mediante estrategias de conservación acordes a las características del área. En este estudio se evaluó el diseño, la gestión y monitoreo de la Reserva Estatal Sierra Monte Negro en el estado de Morelos, México, mediante el uso de variables cualitativas de los estándares abiertos para la conservación y estructuradas de acuerdo con lo sugerido en la evaluación del diseño de CONEVAL. Se encontró que la conceptualización, gestión y monitoreo alcanzaron un grado medianamente satisfactorio, presentando vacíos importantes como la declaratoria de una visión general, la identificación clara y priorización tanto de los objetos de conservación como de las amenazas directas, la ambigüedad de metas, estrategias y objetivos, y la carencia de indicadores para el monitoreo. Los resultados permitirán mejorar la planeación del manejo en las ANP.
Germinación de Ferocactus townsendianus Britton & Rose. con escarificación de semillas sometidas a diferentes tiempos de almacenamiento
Authors: Guillen-Trujillo Ariel , Ávalos-Castro Raúl , Espinoza-Villavicencio José Luis , Ortega-Pérez Ricardo , Palacios-Espinosa Alejandro
Published: February 2021

Se evaluó el efecto del tiempo de almacenamiento y seis tratamientos de escarificación sobre la germinación de semillas de Ferocactus townsendianus. Dos procedimientos a base de ácido sulfúrico (H2SO4), dos con ácido clorhídrico (HCl), uno con agua y uno testigo fueron comparados usando un modelo lineal con mediciones repetidas en el tiempo y contrastes ortogonales. El H2S04 y HCl aumentaron la germinación (P <0.05) en relación con el testigo, en las semillas con 1, 2, 3, 6 y 9 años de almacenamiento, siendo en términos generales mejor el H2S04, este tratamiento también aumentó la germinación (P <0.05) de las semillas con 11 años de almacenamiento. El resto de las semillas no presentó diferencias (P >0.05) entre los procedimientos. El realizado con agua tuvo una germinación nula.
Carbono retenido en biomasa y suelo en bosques de El Salto, Durango, México
Authors: Morales-Inocente Miguel Ángel , Nájera-Luna Juan Abel , Escobedo-Bretado Miguel Ángel , Cruz-Cobos Francisco , Hernández Francisco Javier , Vargas-Larreta Benedicto
Published: February 2021

Los bosques retienen cantidades grandes de CO2 en los principales componentes del ecosistema forestal. El conocimiento sobre esta condición se convierte en una herramienta con potencial para mitigar el cambio climático. Por tanto, este estudio se centra en estimar el carbono retenido en biomasa aérea y suelo en bosques de la región de El Salto, Durango, México. Para ello, información dasométrica y muestras de suelo provenientes de una red de nueve parcelas permanentes de investigación forestal fueron usadas. Los resultados indican la existencia de 135.95 Mg ha-1 de biomasa aérea equivalente a una retención de carbono de 66.42 Mg ha-1. El 95% de carbono se encuentra retenido en la biomasa arbórea y 5% enel sotobosque (hojarasca, necromasa, arbustivas y herbáceas). Por otra parte, el carbono orgánico de suelo estimado es de 187.06 Mg ha-1, 70% se encuentra en los primeros 20 cm de profundidad.
Monitoreo de la antigüedad de hospitales potencialmente requeridos en caso de desastre en México
Authors: Montejano-Castillo Milton , Moreno-Villanueva Mildred
Published: February 2021

Desastres recientes han evidenciado la necesidad de revisar en México la seguridad de la infraestructura crítica del país. En este artículo se exploran las limitantes que un hospital puede tener con el fin de incrementar su seguridad ante desastres de acuerdo con su antigüedad, dada la relativamente reciente formulación de códigos de construcción y lineamientos para hacer los hospitales seguros frente a desastres. Por ello, el año de construcción y el coeficiente de ocupación se tomaron como indicadores para analizar algunos de los hospitales en proceso de certificación en el programa Hospital Seguro en México. Se observa que solo una minoría de los hospitales es de reciente construcción y queen su mayoría estos hospitales están sujetos a una constante expansión física, por lo que se propone un Observatorio para identificar vulnerabilidades asociadas a la antigüedad y saturación física de hospitales, para evaluar la factibilidad de recibir saldos masivos
Destellos arquitectónicos en Aguascalientes
Authors: Ramírez-Villalpando Rocío
Published: February 2021

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道路桥梁工程伸缩缝施工质量技术的控制策略研究
Authors: 朱 鸿章
Published: February 2021

伸缩缝是道路桥梁工程中质量控制的关键，道路桥梁运行中需要承受来自自然环境、车辆等多种外来因素的荷载，导致提前破坏伸缩缝，影响道路桥梁行车舒适性和防水性等各项性能。只有明确伸缩缝在道路桥梁工程中的重要作用、分析影响伸缩缝质量的因素，才能不断优化改进伸缩缝的施工技术和质量，延长道路桥梁施工寿命和行车安全性，推动我国交通行业不断发展。
建筑施工现场文明施工管理探讨
Authors: 田 卫忠
Published: February 2021

近年来，在多方面利好因素的影响下，我国综合国力整体水平得到了显著的提升，为各个领域的发展壮大带来了诸多的机遇，尤其是建筑工程行业的发展势头更加的迅猛。在实施建筑工程施工工作的过程中，在施工现场积极的落实文明施工管理是人类社会发展的必然结果，并且也是建筑工程施工单位最为重要的管理工作内容。不断增强建筑工程施工现场文明施工管理工作整体水平，推动各项施工工作的高效有序开展，切实的优化施工现场施工环境，在推动整个建筑工程行业的未来良好发展方面能够起到积极的影响作用。
Implementasi Program Mini-Seminar Bahasa Inggris Bidang Kesehatan Untuk Program Studi Kesehatan Masyarakat Undikma Mataram
Authors: Kamarudin Kamarudin , Haerazi Haerazi , Sugianto Sugianto , Imran Fathurrahman , Tawali Tawali
Published: February 2021

The implementation of Mini-Seminar of English language for health at social health major of UNDIKMA Mataram aims to improve students’ English language knowledge, capability in Social Health Program of Undikma, especially English for social health both theoretically and practically. This program involved the students of social health in Undikma Mataram. The presenters or the mentors of this program were the English lecturers, and the students majoring in social health who are still active as students as the participants. The program took place in Mandalika University of Education (UNDIKMA), Mataram. The program implemented the method included planning, acting, observing in which through giving the material presentation focusing on some materials such as; English for health, eight parts of speech, modal auxiliaries, mini conversation practice realizing the materials given. The result of the implemented program revealed the high enthusiasm and interest to participate actively within the process of the program given start from the first till the end. The students also gained the increase improvement regarding the capability of using English language expression for social health, vocabulary items, and self-confident. The students also expected the same program may be re-conducted since they showed a high appreciation to the program given in terms of presentation, and guiding within the practice, so that they can enrich and improve their English language communication skill especially English for social health.
ВИКОРИСТАННЯ ЗАСОБІВ ІКТ ДЛЯ ФОРМУВАННЯ НАВЧАЛЬНИХ І КОМУНІКАТИВНИХ СТРАТЕГІЙ ПІД ЧАС ВИВЧЕННЯ ІНОЗЕМНОЇ МОВИ У НЕМОВНИХ ЗАКЛАДАХ ВИЩОЇ ОСВІТИ
Authors: Тадеєва Марія Іванівна , Купчик Лариса Євгенівна , Літвінчук Альона Теофанівна
Published: February 2021

Стаття присвячена проблемам інноваційного розвитку освітнього процесу вивчення іноземної мови засобами інформаційно-комунікаційних технологій. Аналіз нормативно-правової бази, на основі якої здійснюється підготовка здобувачів вищої освіти у немовних закладах вищої освіти, дав змогу визначити актуальність формування іншомовної комунікативної компетентності і здатності до міжкультурної комунікації, зокрема професійної. Охарактеризовано компетентнісний підхід у процесі реалізації іншомовної освіти, що базується на інтеграції ключових компетентностей, покладених в основу формування змістових ліній вивчення мов, які конституюються з мовного, мовленнєвого, лінгвокультурного матеріалу та навчальних і комунікативних стратегій. Здійснено аналіз таких лінгводидактичних категорій, як-от: зміст іншомовної освіти, стратегії навчання, форми і методи організації вивчення іноземних мов. Особливу увагу приділено проблемі формування навчальних стратегій як засобу реалізації неперервної іншомовної освіти, проаналізовано низку існуючих класифікацій стратегій у зарубіжному і вітчизняному освітньому просторі та запропоновано шляхи їх технологізації. Застосування інформаційно-комунікаційних технологій розглядається як основний фактор формування позитивної мотивації та створення ситуації успіху в процесі формування навчальних стратегій, де основними чинниками вважаємо персоналізацію, довіру, доступність, гнучкість, продуктивність й інтерактивність. Окреслено низку ефективних методів навчання іноземних мов з використанням різноманітних інформаційно-комунікаційних технологій, які мають на меті формування сприятливого навчального середовища, підвищення мотиваційного потенціалу іншомовної освіти, активізацію самостійної роботи студентів, розвиток їх критичного мислення шляхом конструювання змісту освіти, що передбачає усвідомлений відбір, опрацювання та засвоєння навчальної інформації.
ГЕЙМДИЗАЙН ОРІЄНТОВАНИЙ ПІДХІД ДО РОЗРОБКИ НАВЧАЛЬНИХ ДИСЦИПЛІН ЗАКЛАДІВ ВИЩОЇ ОСВІТИ
Authors: Лугова Тетяна Анатоліївна
Published: February 2021

У статті обґрунтовується доцільність геймдизайн орієнтованого підходу до розробки дисциплін у закладах вищої освіти (ЗВО), що надалі стане основою для впровадження віртуальних навчально-ігрових середовищ. Показано різницю між поняттями «гейміфікація», «геймдизайн», «Game-based learning». Описано структурні моделі ігродисципліни (технологічна, покрокова, кластерна, прецедентна та змістовно-картографічна), надано їх характеристики, способи імплементації. Визначено етапи та особливості впровадження елементів геймдизайну в навчальних дисциплінах ЗВО. Проаналізовано переваги та недоліки «гейміфікаційної класної кімнати». Представлено схему розподілу елементів навчального контенту за моделлю гри МDA (Механіка – Динаміка – Естетика). Зазначено, що в результаті впровадження геймдизайн-орієнтованого підходу має змінитися погляд на планування, реалізацію та контроль процесу навчання, формування стану «занурення» та ефекту «повної присутності» як з боку викладача-методиста-геймдизайнера, так і здобувача освіти як геймера, який набуває досвіду. Запропоновано методику розробки ігродисципліни, що візуально узагальнено в блок-схемі. Акцентовано, що розробка навчальної дисципліни за принципами та методами геймдизайну перебуває на перетині різних галузей знань та прагне створити синергію методики навчання, ігрової кіберкультури, проєктної діяльності та маркетингу. Основою такого підходу є процес формалізації та структуризації дисципліни в реляційні бази даних та карти знань з подальшою їх творчою трансформацією в ігровий світ. Застосування принципів геймдизайну відкриває нові можливості для реалізації інтегративності, міждисциплінарності, персоналізації та гуманізації навчання. Такий підхід не лише підвищує мотивацію, залучення та інтерес до навчання, посилює позитивні ефекти від вивчення дисципліни, а й кардинально змінює ідеологію навчання, його структуру та поведінку як системи-тьютора.
ЕМОЦІЙНИЙ ШТУЧНИЙ ІНТЕЛЕКТ У ПРОФЕСІЙНІЙ ПІДГОТОВЦІ МАЙБУТНІХ ПСИХОЛОГІВ
Authors: Дерев'янко Світлана Петрівна
Published: February 2021

Статтю присвячено визначенню можливостей застосування знань про емоційний штучний інтелект у процесі викладання курсу «Загальна (когнітивна) психологія» для студентів-психологів. Уточнено визначення поняття «емоційний штучний інтелект»: це інтелектуальні системи, які здатні розпізнавати емоції людини та інтерпретувати їх. Запропоновано комплекс навчальних завдань, спрямованих на вивчення емоційного штучного інтелекту майбутніми психологами: збір наукової інформації про феномени штучного інтелекту та емоційного штучного інтелекту, зображень про емоційний штучний інтелект; аналіз художніх відеофільмів про емоційний штучний інтелект; аналіз комп’ютерних ігор на основі емоційного штучного інтелекту. Проаналізовано результати навчальної діяльності студентів-психологів з вивчення проблематики емоційного штучного інтелекту у формі психологічних есе та мультимедійних творчих проєктів. Виділено типові помилки, які були допущені авторами мультимедійних творчих проєктів (велика кількість слайдів, відсутність чіткого плану, перенасиченість текстом, анімацією). Презентовано накопичені цифрові ресурси з художніми відеофільмами та комп’ютерними іграми на основі емоційного штучного інтелекту. Продемонстровано зміну уявлень студентів після активного збору інформації про емоційний штучний інтелект (стосовно параметра рефлексивності). Визначено, що застосування знань про емоційний штучний інтелект у процесі професійної підготовки майбутніх психологів сприяло реалізації їх рефлексивних та творчих можливостей. Відзначено ряд загальних та фахових компетентностей, які формуються у студентів при вивченні проблематики емоційного штучного інтелекту, зокрема це вміння самостійно збирати та критично аналізувати інформацію щодо феноменів інтелекту, штучного інтелекту, емоційного штучного інтелекту; здатність до професійного самовдосконалення засобом усвідомлення власних емоційних здібностей (здатності розпізнавати емоції, здатності розуміти власні емоції, управляти ними, здатності до емпатії).
IMPLEMENTING ICT INTO LANGUAGE AND LAW CLASSROOM TO DEVELOP LAW STUDENTS’ COMMUNICATIVE COMPETENCE
Authors: Юлінецька Юлія Василівна , Бабій Оксана Юріївна , Гловюк Ірина Василівна , Степаненко Андрій Сергійович , Пашковський Микола Іванович
Published: February 2021

The new context of changes in labour-market requirements calls for development of foreign language communication competence and use of technologies that promote professional life-long learning. Individual characteristics necessary for a professional to integrate into the international socio-cultural environment have recently been challenged particularly because of growing demands to the ability to communicate, be autonomous and responsible for the achieved learning outcomes. A switch of the European higher education content to the competence-based learning contributes to the development of the mentioned characteristics and competence. Further, the National Qualifications Framework of Ukraine (7 Level, Second cycle (Master)) envisages mastering foreign language communicative competence in professional activities. This research addresses the issue of ICT tools application in the development of foreign language communicative competence of law students during individual study. We focus on blended learning model, which is combination of traditional and distance learning approaches. To conduct the research, focus groups were defined and examined; relevant authentic material (the VOTA, HUDOC database, and CURIA case-law database) analysed; and the most appropriate ICT tools established. The department of foreign languages of the National University "Odessa Academy of Law" designed, tested, piloted and uploaded the on-line dictionary "gLAWssary"; developed and recorded the video course "Legal English in Use" which is offered on the university website. The results indicate that the on-line dictionary and video course supplemented by the workbook are powerful tools to promote professional life-long learning and that individual study skills are central to this task. This finding suggests that a thoroughly elaborated course with the relevant ICT tools could be the basis for building foreign language communicative competence of future lawyers.
TRAINING IN DIGITAL ENTREPRENEURSHIP AS A BASIS FOR FORMING THE INTELLECTUAL CAPITAL OF NATION
Authors: Камінський Олег Євгенович , Єрешко Юлія Олександрівна , Кириченко Сергій Олександрович , Тульчинський Ростислав Володимирович
Published: February 2021

Nowadays, digital technologies have a significant impact on how business perceptions change and how new businesses are created. The new technological paradigm harnesses the potential of collaboration and intellectual capital to develop and implement more robust and sustainable entrepreneurial initiatives. However, while a research in digital entrepreneurship is relevant and timely, there is a limited amount of studies on the real impact of digital technology and intellectual capital on entrepreneurship education, although inquiries into digital entrepreneurship theory and practice have drawn more attention in the recent years. Moreover, at the current level of globalization, society, as practice shows (the situation with COVID-19), is not protected from global shocks in the form of chain crises and pandemics. Therefore, digital entrepreneurship is also able to help solve the economic problems caused by the above phenomena. The purpose of the article is to analyse and clarify various aspects of training students of economic specialties in digital entrepreneurship; formulate the tasks that arise while using digital technologies in the economy, and determine the ways to solve them in the education system. Among many forms of digital entrepreneurship training, online courses are one of the strongest trends and they do affect the content and flow of teaching and learning. This article also focuses on better understanding the skills, opportunities, and risks involved in using the online courses as a new way of training entrepreneurship in higher education system. The authors empirically study the problems and drivers for developing such courses as a new pedagogical concept. The results show that such a course is flexible in time and space and can therefore contribute to the accessibility of entrepreneurship education.
Did Modernity End Polyvalence? Some Observations on Tolerance for Ambiguity in Sunnitafsīr
Authors: Coppens Pieter
Published: February 2021

The transition from a polyvalent premodern tafsīr tradition to a more monovalent modern tradition has been noted by several authors. This article tries to more precisely locate this shift through a case study of commentaries on Q. 53:11, on the controversy over whether the Prophet saw God. I intend to show that from the nineteenth century onwards there indeed was a decline of polyvalence in the discussion of this verse, culminating in a univocal choice for an angelic instead of a divine vision in the twentieth century. I claim that one of the main reasons for the decline of polyvalence on this particular issue is the rise of the trend to approach suras as a unity in tafsīr, instead of a more atomistic approach to separate verses which was more common in premodern tafsīr traditions. A claim sometimes made that the decline of polyvalence has to do with the rise of Salafism or ḥadīth-minded scholarship of the likes of Ibn Kathīr thus seems unjustified.
LA CENTRALIDAD ABSOLUTA Y RELATIVA DEL TRABAJO EN ECUADOR. APROXIMACIÓN CUANTITATIVA.
Authors:   Universidad Autónoma del Estado de Hidalgo, Mejía Reyes Carlos , Rea García Fernando ,   Universidad Estatal de Bolívar, Pizarro Hernández Karina ,   Universidad Autónoma del Estado de Hidalgo
Published: February 2021

El presente estudio revisa la importancia del trabajo en la vida de ciudadanos(as) ecuatorianos(as) en el levantamiento de la Encuesta Mundial de Valores 2017-2020. El objetivo es analizar la posición que se le otorga a la actividad frente a distintas esferas de la vida en Ecuador con países latinoamericanos y Unión Europea. A su vez se pone a prueba la tesis del post-materialismo que explica su tendencia a partir de las condiciones materiales inmediatas manifestadas en indicadores macro-económicos. Los hallazgos señalan que Ecuador es un país cuya centralidad del trabajo es de las más altas en las muestras de ambos continentes y se realza como una prioridad frente a otras esferas de la vida. La razón de ello se advierte por la correlación negativa entre menor bienestar y sensación de seguridad material de una población, la preferencia hacia la valoración del trabajo es mayor.
EVALUASI PERFORMA SPIRAL HEAT EXCHANGER HE-201 PADA UNIT DEMONOMERISASI
Authors: Urbaningtyas Herda , Hendrawati Nanik , Choirudin Fanny
Published: February 2021

Heat exchanger adalah proses perpindahan panas fluida dengan fluida lain tanpa terjadi perpindahan massa  didalamnya dan dapat dipergunakan sebagai pemanas. Salah satu tipe alat heat exchanger yang dipakai pada  unit demonomerisasi adalah Spiral heat exchanger. Pada Demonomerisasi di PT Ashahimas Chemical, Spiral heat  exchanger HE – 201 digunakan untuk memanaskan atau meningkatkan suhu slurry demonomer. Spiral heat  exchanger HE-201 tersebut terdapat di unit demonomerisasi pada proses PVC (Polyvinyl Chloride). Hal ini  dilakukan karena slurry yang terproduksi mengalami penurunanan yang disebabkan terbentuknya kerak. Hal  tersebut menyebabkan koefiesien perpindahan panas (Ud) menurun dan mencapai jenuh, oleh karena itu perlu  dilakukan evaluasi untuk mengetahui nilai perpindahan panas dan efisiensi performa alat. Berdasarkan hasil  evaluasi dari perhitungan nilai flouling factor (Rd) pada HE-201 diperoleh Rd actual yang jauh lebih kecil dari Rd  desain dan nilai ΔP aktual sebesar 1,6 Kg/cm2G. Nilai tersebut berada dibawah nilai pressure drop yang diizinkan  yaitu sebesar 2,6 Kg/cm2G. Hal ini menunjukkan bahwa HE-201 pada unit demonomerisasi memiliki kondisi yang  sangat baik, masih dapat beroperasi dan tidak perlu diperbarui.
Comparación de dos suelos para la producción de inoculantes micorrízicos en San Luis Potosí, México
Authors: Mena Echevarría Aracely , Méndez Cortes Heriberto , Ramírez Tobías Hugo M. , Rojas Velázquez Ángel Natanael
Published: February 2021

Resumen
Antecedentes: En la actualidad la tecnología de producción de inóculos micorrízicos ha dado un impulso notable. Sin embargo, es necesario mejorar la calidad de inoculantes con especies micorrízicas eficientes que beneficien a los cultivos.
Objetivos: Identificar la diversidad de Hongos micorrízicos Arbusculares (HMA) de dos suelos de San Luis Potosí y su respuesta en el crecimiento de plantas de S. vulgare y Zea. mays.
Métodos: Se utilizaron dos tipos de suelo (húmedo y semiárido) y plantas trampas de sorgo y maíz. Se evaluó el crecimiento, comportamiento micorrízico y el flujo de CO2 del sustrato.
Resultados y conclusiones: Se identificaron 11 especies de hongos micorrízicos pertenecientes a tres familias del Phyllum Glomeromycota. La familia Glomeraceae fue más abundante seguida de Acaulosporaceae y Claroideo-Glomeraceae. Los cultivos aumentaron su biomasa aérea en el sustrato semiárido y la biomasa radicular en el sustrato húmedo. La mayor tasa de flujo de CO2 se encontró en el sustrato semiárido. El número de esporas fue superior en el sustrato semiárido y el porcentaje e intensidad de colonización fue mayor en el maíz del sustrato semiárido. La mayor diversidad de HMA se replicó en el cultivo trampa de sorgo.
Palabras claves: Ambientes, Micorrizas Arbusculares Vesiculares, Cultivo trampa
Formation Mechanism and Biomedical Applications of Protease-Manipulated Peptide Assemblies
Authors: Jiang Tianyue , Liu Chendan , Xu Xiao , He Bingfang , Mo Ran
Published: February 2021

Exploiting enzyme-catalyzed reactions to manipulate molecular assembly has been considered as an attractive bottom-up nanofabrication approach to developing a variety of nano-, micro-, and macroscale structures. Upon enzymatic catalysis, peptides and their derivatives transform to assemblable building blocks that form ordered architecture by non-covalent interactions. The peptide assemblies with unique characteristics have great potential for applications in bionanotechnology and biomedicine. In this mini review, we describe typical mechanisms of the protease-instructed peptide assembly via bond-cleaving or bond-forming reactions, and outline biomedical applications of the peptide assemblies, such as drug depot, sustained release, controlled release, gelation-regulated cytotoxicity, and matrix construction.
Optimizing a Fed-Batch High-Density Fermentation Process for Medium Chain-Length Poly(3-Hydroxyalkanoates) in Escherichia coli
Authors: Scheel Ryan A. , Ho Truong , Kageyama Yuki , Masisak Jessica , McKenney Seamus , Lundgren Benjamin R. , Nomura Christopher T.
Published: February 2021

Production of medium chain-length poly(3-hydroxyalkanoates) [PHA] polymers with tightly defined compositions is an important area of research to expand the application and improve the properties of these promising biobased and biodegradable materials. PHA polymers with homopolymeric or defined compositions exhibit attractive material properties such as increased flexibility and elasticity relative to poly(3-hydroxybutyrate) [PHB]; however, these polymers are difficult to biosynthesize in native PHA-producing organisms, and there is a paucity of research toward developing high-density cultivation methods while retaining compositional control. In this study, we developed and optimized a fed-batch fermentation process in a stirred tank reactor, beginning with the biosynthesis of poly(3-hydroxydecanoate) [PHD] from decanoic acid by β-oxidation deficient recombinant Escherichia coli LSBJ using glucose as a co-substrate solely for growth. Bacteria were cultured in two stages, a biomass accumulation stage (37°C, pH 7.0) with glucose as the primary carbon source and a PHA biosynthesis stage (30°C, pH 8.0) with co-feeding of glucose and a fatty acid. Through iterative optimizations of semi-defined media composition and glucose feed rate, 6.0 g of decanoic acid was converted to PHD with an 87.5% molar yield (4.54 g L–1). Stepwise increases in the amount of decanoic acid fed during the fermentation correlated with an increase in PHD, resulting in a final decanoic acid feed of 25 g converted to PHD at a yield of 89.4% (20.1 g L–1, 0.42 g L–1 h–1), at which point foaming became uncontrollable. Hexanoic acid, octanoic acid, 10-undecenoic acid, and 10-bromodecanoic acid were all individually supplemented at 20 g each and successfully polymerized with yields ranging from 66.8 to 99.0% (9.24 to 18.2 g L–1). Using this bioreactor strategy, co-fatty acid feeds of octanoic acid/decanoic acid and octanoic acid/10-azidodecanoic acid (8:2 mol ratio each) resulted in the production of their respective copolymers at nearly the same ratio and at high yield, demonstrating that these methods can be used to control PHA copolymer composition.
Phenotypic and Cellular Characteristics of a Stromal Vascular Fraction/Extracellular Matrix Gel Prepared Using Mechanical Shear Force on Human Fat
Authors: Ye Yuan , Zou Jingjiang , Tan Meijun , Hu Kuikui , Jiang Jindou
Published: February 2021

The retention of fat-derived grafts remains a challenge for regenerative medicine. Fat aspirates from patients undergoing liposuction were prepared into standard Coleman fat grafts or further isolated using mechanical shear force to prepare a stromal vascular fraction (SVF)/extracellular matrix (ECM) gel. The retention rate of the SVF/ECM gel was significantly higher than that of the Coleman fat at 3, 14, 28, and 60 days following transplantation on the backs of nude mice. The viscosity of the fat was directly proportional to the shearing force. Although the mechanical isolation did not affect the total number of cells, it significantly decreased the number of living cells. Flow cytometry showed a greater number of mesenchymal stem cells, supra-adventitial (SA)-adipose stromal cells (ASCs), and adipose-derived stem cells but a lower number of endothelial progenitor cells in the SVF/ECM gel than in the Coleman fat. Thus, mechanical isolation of fat can increase the pluripotency of adipocytes, which can improve graft retention in cell therapy.
Primary Cilia Formation Does Not Rely on WNT/β-Catenin Signaling
Authors: Bernatik Ondrej , Paclikova Petra , Kotrbova Anna , Bryja Vitezslav , Cajanek Lukas
Published: February 2021

Primary cilia act as crucial regulators of embryo development and tissue homeostasis. They are instrumental for modulation of several signaling pathways, including Hedgehog, WNT, and TGF-β. However, gaps exist in our understanding of how cilia formation and function is regulated. Recent work has implicated WNT/β-catenin signaling pathway in the regulation of ciliogenesis, yet the results are conflicting. One model suggests that WNT/β-catenin signaling negatively regulates cilia formation, possibly via effects on cell cycle. In contrast, second model proposes a positive role of WNT/β-catenin signaling on cilia formation, mediated by the re-arrangement of centriolar satellites in response to phosphorylation of the key component of WNT/β-catenin pathway, β-catenin. To clarify these discrepancies, we investigated possible regulation of primary cilia by the WNT/β-catenin pathway in cell lines (RPE-1, NIH3T3, and HEK293) commonly used to study ciliogenesis. We used WNT3a to activate or LGK974 to block the pathway, and examined initiation of ciliogenesis, cilium length, and percentage of ciliated cells. We show that the treatment by WNT3a has no- or lesser inhibitory effect on cilia formation. Importantly, the inhibition of secretion of endogenous WNT ligands using LGK974 blocks WNT signaling but does not affect ciliogenesis. Finally, using knock-out cells for key WNT pathway components, namely DVL1/2/3, LRP5/6, or AXIN1/2 we show that neither activation nor deactivation of the WNT/β-catenin pathway affects the process of ciliogenesis. These results suggest that WNT/β-catenin-mediated signaling is not generally required for efficient cilia formation. In fact, activation of the WNT/β-catenin pathway in some systems seems to moderately suppress ciliogenesis.
TRPM3 in Brain (Patho)Physiology
Authors: Held Katharina , Tóth Balázs István
Published: February 2021

Already for centuries, humankind is driven to understand the physiological and pathological mechanisms that occur in our brains. Today, we know that ion channels play an essential role in the regulation of neural processes and control many functions of the central nervous system. Ion channels present a diverse group of membrane-spanning proteins that allow ions to penetrate the insulating cell membrane upon opening of their channel pores. This regulated ion permeation results in different electrical and chemical signals that are necessary to maintain physiological excitatory and inhibitory processes in the brain. Therefore, it is no surprise that disturbances in the functions of cerebral ion channels can result in a plethora of neurological disorders, which present a tremendous health care burden for our current society. The identification of ion channel-related brain disorders also fuel the research into the roles of ion channel proteins in various brain states. In the last decade, mounting evidence has been collected that indicates a pivotal role for transient receptor potential (TRP) ion channels in the development and various physiological functions of the central nervous system. For instance, TRP channels modulate neurite growth, synaptic plasticity and integration, and are required for neuronal survival. Moreover, TRP channels are involved in numerous neurological disorders. TRPM3 belongs to the melastatin subfamily of TRP channels and represents a non-selective cation channel that can be activated by several different stimuli, including the neurosteroid pregnenolone sulfate, osmotic pressures and heat. The channel is best known as a peripheral nociceptive ion channel that participates in heat sensation. However, recent research identifies TRPM3 as an emerging new player in the brain. In this review, we summarize the available data regarding the roles of TRPM3 in the brain, and correlate these data with the neuropathological processes in which this ion channel may be involved.
In vitro Modeling of Embryonal Tumors
Authors: Custers Lars , Paassen Irene , Drost Jarno
Published: February 2021

A subset of pediatric tumors affects very young children and are thought to arise during fetal life. A common theme is that these embryonal tumors hijack developmental programs, causing a block in differentiation and, as a consequence, unrestricted proliferation. Embryonal tumors, therefore typically maintain an embryonic gene signature not found in their differentiated progeny. Still, the processes underpinning malignant transformation remain largely unknown, which is hampering therapeutic innovation. To gain more insight into these processes, in vitro and in vivo research models are indispensable. However, embryonic development is an extremely dynamic process with continuously changing cellular identities, making it challenging to define cells-of-origin. This is crucial for the development of representative models, as targeting the wrong cell or targeting a cell within an incorrect developmental time window can result in completely different phenotypes. Recent innovations in in vitro cell models may provide more versatile platforms to study embryonal tumors in a scalable manner. In this review, we outline different in vitro models that can be explored to study embryonal tumorigenesis and for therapy development.
Dysregulation of miR-138-5p/RPS6KA1-AP2M1 Is Associated With Poor Prognosis in AML
Authors: Yu Dong-Hu , Chen Chen , Liu Xiao-Ping , Yao Jie , Li Sheng , Ruan Xiao-Lan
Published: February 2021

Acute myeloid leukemia (AML) is a malignant disease of hematopoietic stem/progenitor cells, and most AML patients are in a severe state. Internal tandem duplication mutations in FLT3 gene (FLT3-ITD) detected in AML stem cells account for 20–30 percent of AML patients. In this study, we attempted to study the impact of the interaction of FLT3-ITD mutation and the CXCL12/CXCR4 axis in AML, and the possible mechanisms caused by the impact by bioinformatics. Gene set variation analysis (GSVA) revealed that the PI3K-Akt-mTOR pathway positively correlated with the status of FLT3-ITD mutation. Multiple survival analyses were performed on TCGA-AML to screen the prognostic-related genes, and RPS6KA1 and AP2M1 are powerful prognostic candidates for overall survival in AML. WGCNA, KEGG/GO analysis, and the functional roles of RPS6KA1 and AP2M1 in AML were clarified by correlation analysis. We found that the expression levels of RPS6KA1 and AP2M1 were significantly associated with chemoresistance of AML, and the CXCL12/CXCR4 axis would regulate RPS6KA1/AP2M1 expression. Besides, miR-138-5p, regulated by the CXCL12/CXCR4 axis, was the common miRNA target of RPS6KA1 and AP2M1. Taken together, the interaction of FLT3-ITD mutation and the CXCL12/CXCR4 axis activated the PI3K-Akt-mTOR pathway, and the increased expression of RPS6KA1 and AP2M1 caused by hsa-miR-138-5p downregulation regulates the multi-resistance gene expression leading to drug indications.
Lysosomes and Cancer Progression: A Malignant Liaison
Authors: Machado Eda R. , Annunziata Ida , van de Vlekkert Diantha , Grosveld Gerard C. , d’Azzo Alessandra
Published: February 2021

During primary tumorigenesis isolated cancer cells may undergo genetic or epigenetic changes that render them responsive to additional intrinsic or extrinsic cues, so that they enter a transitional state and eventually acquire an aggressive, metastatic phenotype. Among these changes is the alteration of the cell metabolic/catabolic machinery that creates the most permissive conditions for invasion, dissemination, and survival. The lysosomal system has emerged as a crucial player in this malignant transformation, making this system a potential therapeutic target in cancer. By virtue of their ubiquitous distribution in mammalian cells, their multifaced activities that control catabolic and anabolic processes, and their interplay with other organelles and the plasma membrane (PM), lysosomes function as platforms for inter- and intracellular communication. This is due to their capacity to adapt and sense nutrient availability, to spatially segregate specific functions depending on their position, to fuse with other compartments and with the PM, and to engage in membrane contact sites (MCS) with other organelles. Here we review the latest advances in our understanding of the role of the lysosomal system in cancer progression. We focus on how changes in lysosomal nutrient sensing, as well as lysosomal positioning, exocytosis, and fusion perturb the communication between tumor cells themselves and between tumor cells and their microenvironment. Finally, we describe the potential impact of MCS between lysosomes and other organelles in propelling cancer growth and spread.
Sensing and Responding of Cardiomyocytes to Changes of Tissue Stiffness in the Diseased Heart
Authors: Münch Juliane , Abdelilah-Seyfried Salim
Published: February 2021

Cardiomyocytes are permanently exposed to mechanical stimulation due to cardiac contractility. Passive myocardial stiffness is a crucial factor, which defines the physiological ventricular compliance and volume of diastolic filling with blood. Heart diseases often present with increased myocardial stiffness, for instance when fibrotic changes modify the composition of the cardiac extracellular matrix (ECM). Consequently, the ventricle loses its compliance, and the diastolic blood volume is reduced. Recent advances in the field of cardiac mechanobiology revealed that disease-related environmental stiffness changes cause severe alterations in cardiomyocyte cellular behavior and function. Here, we review the molecular mechanotransduction pathways that enable cardiomyocytes to sense stiffness changes and translate those into an altered gene expression. We will also summarize current knowledge about when myocardial stiffness increases in the diseased heart. Sophisticated in vitro studies revealed functional changes, when cardiomyocytes faced a stiffer matrix. Finally, we will highlight recent studies that described modulations of cardiac stiffness and thus myocardial performance in vivo. Mechanobiology research is just at the cusp of systematic investigations related to mechanical changes in the diseased heart but what is known already makes way for new therapeutic approaches in regenerative biology.
Membrane Transport Proteins in Osteoclasts: The Ins and Outs
Authors: Ribet Amy B. P. , Ng Pei Ying , Pavlos Nathan J.
Published: February 2021

During bone resorption, the osteoclast must sustain an extraordinarily low pH environment, withstand immense ionic pressures, and coordinate nutrient and waste exchange across its membrane to sustain its unique structural and functional polarity. To achieve this, osteoclasts are equipped with an elaborate set of membrane transport proteins (pumps, transporters and channels) that serve as molecular ‘gatekeepers’ to regulate the bilateral exchange of ions, amino acids, metabolites and macromolecules across the ruffled border and basolateral domains. Whereas the importance of the vacuolar-ATPase proton pump and chloride voltage-gated channel 7 in osteoclasts has long been established, comparatively little is known about the contributions of other membrane transport proteins, including those categorized as secondary active transporters. In this Special Issue review, we provide a contemporary update on the ‘ins and outs’ of membrane transport proteins implicated in osteoclast differentiation, function and bone homeostasis and discuss their therapeutic potential for the treatment of metabolic bone diseases.
Lanthanide-Doped Upconversion-Linked Immunosorbent Assay for the Sensitive Detection of Carbohydrate Antigen 19-9
Authors: Zhou Chaohui , Chu Zhongyun , Hou Wenyue , Wang Xiuying
Published: February 2021

Lanthanide-doped upconversion nanoparticles (UCNPs) have attracted considerable attention in detection of biological analytes and bioimaging owing to their superior optical properties, including high photochemical stability, sharp emission bandwidth, large anti-Stokes shifts, and low toxicity. In this work, we fabricated UCNP-linked immunosorbent assay (ULISA) for the sensitive detection of carbohydrate antigen 19-9 (CA19-9). The design is based on amino-functionalized SiO2-coated Gd-doped NaYF4:Yb3+,Er3+ upconversion nanoparticles (UCNPs@SiO2-NH2) as a direct background-free luminescent reporter; a secondary anti-IgG antibody (Ab2) was conjugated to the surface of UCNPs@SiO2-NH2 (UCNP-Ab2), and UCNP-Ab2 was used for specific targeting of CA19-9. The UCNPs were well characterized by TEM, SEM, XRD, FT-IR, and UV-vis. The detection process was similar to enzyme-linked immunosorbent assay (ELISA). UCNPs were used as signal transducer to replace the color compounds for an enzyme-mediated signal amplification step. An anti-CA19-9 primary antibody (Ab1) was fixed for capturing the CA19-9, and the fluorescence signal was obtained from the specific immunoreaction between UCNP-Ab2 and CA19-9. Under optimum conditions, this ULISA shows sensitive detection of CA19-9 with a dynamic range of 5–2,000 U/ml. The ULISA system shows higher detection sensitivity and wider detection range compared with the traditional ELISA for CA19-9 detection. This strategy using UCNPs as signal transducer may pave a new avenue for the exploration of rare doped UCNPs in ELISA assay for clinical applications in the future.
Phylogeography of Human and Animal Coxiella burnetii Strains: Genetic Fingerprinting of Q Fever in Belgium
Authors: Tomaiuolo Sara , Boarbi Samira , Fancello Tiziano , Michel Patrick , Desqueper Damien , Grégoire Fabien , Callens Jozefien , Fretin David , Devriendt Bert , Cox Eric , Mori Marcella
Published: February 2021

Q fever is a zoonotic disease caused by the bacteriaCoxiella burnetii.Domestic ruminants are the primary source for human infection, and the identification of likely contamination routes from the reservoir animals the critical point to implement control programs. This study shows that Q fever is detected in Belgium in abortion of cattle, goat and sheep at a different degree of apparent prevalence (1.93%, 9.19%, and 5.50%, respectively). In addition, and for the first time, it is detected in abortion of alpaca (Vicugna pacos), raising questions on the role of these animals as reservoirs. To determine the relationship between animal and human strains, Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) (n=146), Single-Nucleotide Polymorphism (SNP) (n=92) and Whole Genome Sequencing (WGS) (n=4) methods were used to characterize samples/strains during 2009-2019. Three MLVA clusters (A, B, C) subdivided in 23 subclusters (A1-A12, B1-B8, C1-C3) and 3 SNP types (SNP1, SNP2, SNP6) were identified. The SNP2 type/MLVA cluster A was the most abundant and dispersed genotype over the entire territory, but it seemed not responsible for human cases, as it was only present in animal samples. The SNP1/MLVA B and SNP6/MLVA C clusters were mostly found in small ruminant and human samples, with the rare possibility of spillovers in cattle. SNP1/MLVA B cluster was present in all Belgian areas, while the SNP6/MLVA C cluster appeared more concentrated in the Western provinces. A broad analysis of European MLVA profiles confirmed the host-species distribution described for Belgian samples.In silicogenotyping (WGS) further identified the spacer types and the genomic groups ofC. burnetiiBelgian strains: cattle and goat SNP2/MLVA A isolates belonged to ST61 and genomic group III, while the goat SNP1/MLVA B strain was classified as ST33 and genomic group II. In conclusion, Q fever is widespread in all Belgian domestic ruminants and in alpaca. We determined that the public health risk in Belgium is likely linked to specific genomic groups (SNP1/MLVA B and SNP6/MLVA C) mostly found in small ruminant strains. Considering the concordance between Belgian and European results, these considerations could be extended to other European countries.
The Hypothetical Inclusion Membrane Protein CPSIT_0846 Regulates Mitochondrial-Mediated Host Cell Apoptosis via the ERK/JNK Signaling Pathway
Authors: Tang Ting , Wu Haiying , Chen Xi , Chen Li , Liu Luyao , Li Zhongyu , Bai Qinqin , Chen Yuyu , Chen Lili
Published: February 2021

Chlamydia psittaci is an important zoonotic factor associated with human and animal atypical pneumonia. Resisting host cell apoptosis is central to sustaining Chlamydia infection in vivo. Chlamydia can secrete inclusion membrane proteins (Incs) that play important roles in their development cycle and pathogenesis. CPSIT_0846 is an Inc protein in C. psittaci identified by our team in previous work. In the current study, we investigated the regulatory role of CPSIT_0846 in HeLa cell apoptosis, and explored potential mechanisms. The results showed that HeLa cells treated with CPSIT_0846 contained fewer apoptotic bodies and exhibited a lower apoptotic rate than untreated cells either with Hoechst 33258 fluorescence staining or flow cytometry with or without induction by staurosporine (STS). CPSIT_0846 could increase the phosphorylation of the extracellular signal-regulated kinases 1/2 (ERK1/2) or stress-activated protein kinases/c-Jun amino-terminal kinases (SAPK/JNK) signaling pathways, and the Bcl-2 associated X protein (Bax)/B cell lymphoma 2 (Bcl-2) ratio, levels of cleaved caspase-3/9 and cleaved Poly-ADP-ribose polymerase (PARP) were significantly up-regulated following inhibition of ERK1/2 or SAPK/JNK pathways with U0126 or SP600125. After carbonyl cyanide 3-chlorophenylhydrazone (CCCP) treatment, the mitochondrial membrane potential (MMP) of cells was significantly decreased in control group, but stable in the CPSIT_0846 treated one, and less cytochrome c (Cyt.c) was released into the cytoplasm. Inhibition of the ERK1/2 or SAPK/JNK pathway significantly decreased the JC-1 red-green fluorescence signal, and promoted Cyt.c discharge into the cytoplasm in HeLa cells treated with CPSIT_0846. In conclusion, CPSIT_0846 can regulate mitochondrial pathway-mediated apoptosis in HeLa cells by activating the ERK/JNK signaling pathway.
Deletion of Glutamine Synthetase Gene Disrupts the Survivability and Infectivity of Leishmania donovani
Authors: Kumar Vinay , Ghosh Sanhita , Roy Kamalika , Pal Chiranjib , Singh Sushma
Published: February 2021

Glutamine synthetase (GS) is one of the most important metabolic enzymes which catalyzes ligation of glutamate and ammonia to form glutamine. Previous studies from our lab had revealed significant differences in parasite and host GS enzyme which warranted us to further work on its relevance in parasite. To analyze glutamine synthetase function in Leishmania, we generated GS overexpressors and knockout mutants and evaluated their ability to grow in vitro in monocyte differentiated macrophage and in vivo by infections in BALB/c mice. GS knocked out strain showed significant growth retardation with delayed cell cycle progression and morphological alteration. Null mutants exhibited attenuated infectivity both in in vitro and in vivo experiments and the effect was reverted back when infected with GS complemented parasites. This indicated that the alterations in phenotype observed were indeed due to GS knockout. GS knockout also made the parasite increasingly sensitive to Miltefosine. Detailed investigation of mode of parasite death upon Miltefosine treatment by dual staining with Annexin-V conjugated FITC and propidium iodide, pointed towards apoptotic or necrotic mode of cell death. This is the first report to confirm that GS is essential for the survivability and infectivity of Leishmania donovani, and can be exploited as a potential drug-target.