bluuebunny/crossref_metadata_2025 huggingface dataset:
A Computer Vision Application for Measuring the Deflection in a Two-dimensional View of Reinforced Concrete Beams
Authors:   University of San Jose-Recoletos, Cebu City, Philippines, Piedad Eduardo Jr. , Carpio Barne Roxette ,   Mindanao State University-Iligan Institute of Technology, Iligan City, Philippines, Sanchez Kristine ,   Mindanao State University-Iligan Institute of Technology, Iligan City, Philippines, Jabian Marven ,   Mindanao State University-Iligan Institute of Technology, Iligan City, Philippines
Published: June 2021

A novel computer vision application is developed to measure the deflection of two-dimensional (2D) reinforced concrete structural members. Eight beam samples, with dimensions of 160 mm x 150 mm x 1400 mm are loaded and simulated under a four-point loading test until failure using a reaction framework machine. A camera is fixed at the center front view of the concrete beams to capture the deflection of the samples while testing. In each test, a dial indicator is installed and the maximum deflection is manually recorded. Based on the results, the maximum deflection values recorded by the proposed application obtained an average error of 18.38 % when compared to the manual measured results. This indicates that computer vision-based application can provide a beam-wide scale deflection performance, compared to the traditional point-based deflection reading. This study paves a new possibility of aiding manual measurements of concrete beams and all other structural studies.
Performance Evaluation of a Rice Hull-fueled Cabinet Food Dryer
Authors:   Visayas State University, Baybay City, Leyte, Philippines, Guanco Jerry , Casinillo Leomarich ,   Visayas State University, Baybay City, Leyte, Philippines
Published: June 2021

This study was conducted to determine the technical performance of a rice hull-fueled cabinet food dryer. Data gathered from the drying performance in terms of drying behavior, drying rate, and dryer heat utilization efficiency using cassava chips was analyzed through an experimental design. The study also evaluated fuel consumption, thermal conversion efficiency, energy use efficiency, heat utilization efficiency, temperature distribution and economic performance. Results indicated that the dryer's average fuel consumption and the thermal conversion efficiency is 11.19 kg/hr and 6.17%. While the energy use efficiency is 41.51%, and the dryer efficiency is 11.79 %. The moisture reduction of the rice hull-fueled dryer is 8.156 kg/hr, and the heat utilization efficiency is 41.51%. The break-even of using the dryer is ₱ 4.988/kg; if rented out at ₱ 59.86/hr, the payback period is 0.988 batch/hr.
A motivação empreendedora dos jovens e sua relevância para o desenvolvimento socioeconômico em São Luís
Authors: Silva Valkiria , Oliveira Willian , Fonseca e Fonseca Paulo
Published: June 2021

O artigo possui como problema: qual a motivação empreendedora dos jovens empresários ludovicenses. Sendo assim, o objetivo foi de levantar as principais motivações que levaram os jovens empresários ludovicenses a empreender. Para obtenção dos dados, foi aplicado um formulário eletrônico em uma IES entre jovens universitários. Quanto à abordagem a pesquisa foi quantitativa, haja vista ter se analisado de forma quantitativa o nível de concordância e discordância com sete variáveis apontadas pela literatura como as principais motivações empreendedoras junto a jovens empresários maranhenses. O resultado da pesquisa apontou que a variável que mais motiva os jovens a empreenderem dentre as sete analisadas foi alcançar a independência financeira.
European High Frequency Radar network governance
Authors: Rubio Anna , Reyes Emma , Mantovani Carlo , Corgnati Lorenzo , Lorente Pablo , Solabarrieta Lohitzune , Mader Julien , Fernandez Vicente , Pouliquen Sylvie , Novellino Antonio , Karstensen Johannes , Peithakis George
Published: June 2021

This report describes the governance of the European HF radar network including: the landscape of the Ocean observation networks and infrastructures, the role and links between operators of observational systems and stakeholders, the role and activities of the EuroGOOS HF radar Task Team in building a sound community strategy, the roadmap of the community with current achievements and future work lines.
¿Por qué importa aprender álgebra elemental?
Authors: Márquez Elías Miguel Ángel
Published: June 2021

La educación media superior es un enclave formativo fundamental en la instrucción de millones de individuos, pues impulsa el acceso a la educación superior. Las matemáticas que ahí se estudian guardan una relación funcional y más que inmediata con las matemáticas con que se formarán profesionistas y, en este contexto, el álgebra elemental juega un papel educativo apreciable, al estimular la imaginación mediante su simbología y las ideas que ésta encierra. En este artículo se advierte su relevancia y competencia en el mundo profesional, así como su necesidad en la vida ordinaria como estímulo cultural, el cual favorezca un entendimiento conveniente del mundo que puede empezar a resolverse de manera adecuada desde el aula, usando herramientas tecnológicas ahora a la mano, creando problemas estimulantes que desarrollen la creatividad a través del uso práctico del álgebra, como lo hacen físicos e ingenieros.
El conocimiento especializado del profesor de matemáticas (MTSK)
Authors: Adame Esparza Alejandra , De Lira Lozano Erick del Refugio
Published: June 2021

En la actualidad, la labor docente en la educación media superior, específicamente en el área de matemáticas, es realizada por una gran variedad de profesionistas que cuentan con diversos perfiles dentro de su formación inicial. La problemática recae en que gran parte de estos profesionistas no han sido formados con un perfil docente para cubrir los requerimientos demandados por este nivel. Es por ello que la matemática educativa, en su afán de contribuir al desarrollo profesional de los docentes en activo, propone el modelo del conocimiento especializado del profesor de matemáticas (Mathematics Teacher’s Specialised Knowledge, mtsk), el cual pretende hacer reflexionar acerca de los conocimientos que un profesor de matemáticas debiera poseer para desempeñar esta labor de manera especializada y en beneficio de los estudiantes. La presentación de este modelo es lo que atañe el presente artículo.
Revaloración de enfoques tradicionales en la enseñanza de lenguas adicionales
Authors: Correa Larios Olivia
Published: June 2021

En el presente trabajo se analizan algunas tensiones latentes entre el enfoque tradicional de enseñanza de lenguas de “gramática-traducción” y el enfoque “comunicativo por competencias” en los cursos de lenguas adicionales al inglés (italiano, alemán o francés), impartidos en el programa de Licenciatura en Lenguas Extranjeras (LILEX) de la Universidad Autónoma de Zacatecas (UAZ), con el fin de explorar las oportunidades de creatividad en la actividad docente, esto a través de propuestas didácticas de inclusión de literatura y traducción como actividades complementarias al uso de los manuales didácticos estandarizados que no siempre se ajustan a las necesidades  educativas del referido contexto de enseñanza y aprendizaje.
El impacto de la tutoría a través del acompañamiento longitudinal.
Authors: Báez Ruiz Esparza Carolina Monserrat , Landeros Romo Angélica Rocío , Sánchez Navarro María de los Ángeles
Published: June 2021

Se presenta el contexto general de desarrollo de la tutoría longitudinal en el Centro de Educación Media (CEM) de la Universidad Autónoma de Aguascalientes (UAA). En este sentido, se destaca la relevancia del perfil del tutor longitudinal para llevar a cabo la función de seguimiento al estudiante tanto en lo académico como en lo personal, familiar y social. Finalmente, se comparte la experiencia en la incorporación de la estrategia de plan de vida del estudiante, a partir de la propuesta de la Subsecretaría de Educación Media Superior de la Secretaría de Educación Pública (SEP), para la implementación de las tutorías longitudinales en el mismo centro, la cual puede ser de utilidad para los tutores que laboran en la educación media superior.
La tutoría en la elección de carrera
Authors: Barba Macías Jesús Francisco
Published: June 2021

Uno de los principales problemas que enfrentan las universidades es la deserción estudiantil durante los primeros semestres de la carrera, debido a la falta de una adecuada orientación vocacional, previa a la elección de la misma. De acuerdo con los datos publicados por dos instituciones de educación superior sobre la falta de discernimiento vocacional de los estudiantes como una de las principales causas de deserción, se propone analizar la actividad tutorial y la función del tutor en la educación media superior como una estrategia para orientar vocacionalmente a los estudiantes, en fomento a su adecuada elección de carrera.
El aprendizaje significativo en el contexto de la educación musical superior
Authors: Capistrán Gracia Raúl W.
Published: June 2021

En este artículo, el autor presenta una reflexión sobre las áreas de oportunidad que algunos estudiantes reflejan al momento de interrelacionar los objetivos y contenidos que integran las diversas asignaturas del plan de estudios (PE) de la Licenciatura en Música de la Universidad Autónoma de Aguascalientes (UAA). Para subsanarlas, propone que los docentes se apoyen en la teoría del aprendizaje significativo de David Ausubel, con el propósito de fomentar procesos de enseñanza-aprendizaje que promuevan el desarrollo del pensamiento crítico y reflexivo e impulsen una formación superior integral. Por lo anterior, se incluye una descripción panorámica de la teoría del aprendizaje significativo, así como algunas estrategias e ideas para impulsarlo.
Eğimli ve Kurak Koşullarda Bir Arazi Toplulaştırma Sahasının Çölleşme Potansiyelinin Fraktal Analizle Araştırılması
Authors: ALTUNSU Murat , OGUZ İrfan , KOÇYİĞİT Rasim
Published: June 2021

Fraktal teori, toprak sistemlerinin performansını daha iyi anlamak için toprak yapı dinamiklerini tanımlayan yararlı bir araç haline gelmiştir. Arazi eğim değişiklikleri toprağın fiziksel, kimyasal ve biyolojik özelliklerini önemli ölçüde etkiler. Bununla birlikte, farklı eğimlerde yer alan toprak tabakalarına ait fraktal özellikleri hakkında sınırlı bilgi mevcuttur. Modern istatistiksel çalışmalar topraklardaki heterojenliği tanımlayarak farklı toprakların çölleşme eğilimlerini karşılaştırmaya imkan vermektedir. Bu çalışmada, dört farklı eğim grubunda (% 0-2, % 3-6, % 7-12, >% 12) on altı adet toprak değişkeni için toprak profili boyunca çölleşme eğilimini tanımlamak üzere fraktal analiz istatistiksel teknikleri kullanılmıştır. Bu amaçla, Aksaray iline bağlı Ortaköy ilçesi arazi toplulaştırma proje sahasından toplam 1808 toprak örneği toplanmıştır. Çalışmada toprak tekstürü, doygunluk, pH, EC, tuz, kireç, Ca + Mg, Na, SAR, ESP, B, Kil Oranı I, Kil Oranı II ve Kil Oranı III içerikleri dikkate alınmıştır. Fraktal katsayıya göre, pH, tuz, kireç, silt, Ca + Mg, Na, SAR ve ESP önemli farklılıklar göstermiştir. Genel olarak, eğim arttıkça fraktal katsayı değerleri düşmüştür. Fraktal yaklaşımın toprakların heterojenliğini belirlemede uygun ve önerilebilir olduğu belirlenmiştir.
The complex karyotype landscape in chronic lymphocytic leukemia allows the refinement of the risk of Richter syndrome transformation
Authors: Visentin Andrea , Bonaldi Laura , Rigolin Gian Matteo , Mauro Francesca Romana , Martines Annalisa , Frezzato Federica , Pravato Stefano , Gargarella Leila Romano , Bardi Maria Antonella , Cavallari Maurizio , Volta Eleonora , Cavazzini Francesco , Nanni Mauro , Facco Monica , Piazza Francesco , Guarini Anna , Foà Robin , Semenzato Gianpietro , Cuneo Antonio , Trentin Livio
Published: June 2021

Complex karyotype (CK) at chronic lymphocytic leukemia (CLL) diagnosis is a negative biomarker of adverse outcome. Since the impact of CK and its subtypes, namely type-2 CK (CK with major structural abnormalities) or high-CK (CK with ≥5 chromosome abnormalities), on the risk of developing Richter syndrome (RS) is unknown, we carried out a multicenter real-life retrospective study to test its prognostic impact. Among 540 CLL patients, 107 harbored a CK at CLL diagnosis, 78 were classified as CK2 and 52 as high-CK. Twenty-eight patients developed RS during a median follow-up of 6.7 years. At the time of CLL diagnosis, CK2 and high-CK were more common and predicted the highest risk of RS transformation, together with advanced Binet stage, unmutated (U)-IGHV, 11q-, and TP53 abnormalities. We integrated these variables into a hierarchical model: high-CK and/or CK2 patients showed a 10-year time to RS (TTRS) of 31%; U-IGHV/11q- /TP53 abnormalities/Binet stage B-C patients had a 10-year TTRS of 12%; mutated (M)-IGHV without CK and TP53 disruption a 10-year TTRS of 3% (P<0.0001). We herein demonstrate that CK landscape at CLL diagnosis allows the risk of RS transformation to be refined and we recapitulated clinico-biological variables into a prognostic model.
Formação de professores para o ensino remoto: Uma intervenção realizada no município de Riacho de Santana/RN
Authors: Fernandes Anábia Vitória , Aquino Adelmo Artur de , Oliveira Marcelo Tavares de , Farias Antonio Diego Silva , Lavor Otavio Paulino
Published: June 2021

O ano de 2020 se encontra marcado por uma intensa política de isolamento social em decorrência do COVID-19. Contudo o desenvolvimento tecnológico tem contribuído para os diversos setores, entre estes o ensino, através das Tecnologias de Informação e Comunicação (TICs), influenciando positivamente na aprendizagem e fortalecendo os processos de ensino, proporcionando aos indivíduos distantes do ambiente educacional, uma possibilidade de obter conhecimento, assim como aos que desejam uma nova qualificação, uma alternativa de acesso a diversos conteúdos de forma remota. Abordando a utilização das ferramentas tecnológicas, a presente pesquisa visa a melhoria qualitativa do ensino e da aprendizagem, mediante ao distânciamento social, por meio de uma qualificação de docentes. Os recursos utilizados foram os softwares Vegas Pro 17, OBS Studio e 4k download, que auxiliam na gravação e edição de vídeoaula, contribuindo para que a educação a distância ocorra de forma mais eficaz. Com a formação os docentes foram capazes de planejar, gravar e editar videoaulas que seram compartilhadas aos discentes, atuando como encaminhamentos metodológicos mais interativos e produtivos.
Organização de produtos alimentícios em prateleiras de supermercado em Muritiba – Bahia
Authors: Ferreira Daliane de Oliveira Santos Conceição , Costa Edvaldo Nascimento , Rodrigues Estefânia Prates , Silveira Paulo Túlio de Souza
Published: June 2021

O varejo do Brasil sofre alterações a cada dia com a intenção de atender as demandas dos consumidores. Os supermercados se reinventam a cada dia para que as necessidades visuais da organização das prateleiras sejam eficientes para os clientes e com isso aumentar a sua lucratividade.  O estudo foi realizado em um supermercado da cidade de Muritiba – Bahia, entre de setembro a novembro de 2019. A avaliação foi realizada de forma qualitativa, através da observação do dia a dia de um supermercado.  Para a discussão das observações foi realizada busca de dados para se realizar uma discussão comparativa referente a organização e rotinas existentes no setor. Desse modo, possibilitando o melhor entendimento da natureza das variáveis a fim de desenvolver argumentos e propor soluções para a problemática. Com a observação foi verificar que os supermercados necessitam de melhorias para que os clientes possam ser atendidos de forma adequada e eficiente. Destinar um único colaborador responsável pelo setor de compras, além de colaboradores que confiram os lotes, a validade e se os produtos possuem alguma avaria, são imprescindíveis para garantir a qualidade dos produtos oferecidos, bem como garantir um menor retrabalho na organização das gôndolas do supermercado.
Alterações bucais em pacientes submetidos a tratamento quimioterápico
Authors: Silva Francielly Karla Vieira da , Fursel Keven de Assis , Oliveira Neto José Lopes de , Villas Boas Myllena Isabel Bueno , Pereira Claudio Maranhão
Published: June 2021

O tratamento oncológico quimioterápico invariavelmente é responsável por desencadear alterações bucais advindas da diminuição da resposta imunológica do paciente. A quimioterapia é uma das mais utilizadas e eficazes formas de tratamento oncológico, contudo desencadeia inúmeros efeitos colaterais em todos os sistemas orgânicos, inclusive em cavidade bucal. Dentre eles destaca-se a mucosite, uma das causas de maior desconforto para o paciente, responsável também por uma instabilidade em sua qualidade de vida. É essencial que o paciente com câncer procure um cirurgião dentista para que haja uma prevenção, tratamento e acompanhamento destas alterações objetivando uma melhora na qualidade de vida antes, durante e após quimioterapia ou qualquer outro tratamento oncológico. Com isso é de suma importância determinar as principais alterações bucais relacionadas ao tratamento quimioterápico, assim como avaliar a prevalência de cada alteração e seus efeitos tóxicos.
Genetic evidence from embryos suggests a new species of skate related to Bathyraja parmifera (Rajiformes: Arhynchobatidae) in the Bering Sea
Authors: Spies I , Orr JW , Stevenson DE , Goddard P , Hoff GR , Guthridge J , Rooper CN
Published: June 2021

Several new species of oviparous skates of the genus Bathyraja have been identified over the past 2 decades, yet it is possible that a complete understanding of species diversity among this group has not been achieved. We used genetics and morphology to screen for the presence of species-level differentiation among embryos from nursery areas in the eastern Bering Sea that were initially identified as the Alaska skate B. parmifera. A sample of 57 embryos from Pervenets Canyon differed significantly at single nucleotide polymorphism loci from 297 other B. parmifera samples, and differences were on the order of those observed between B. parmifera and the leopard skate B. panthera. This sample of embryos was similar to B. parmifera at the cytochrome c oxidase subunit I (COI) mitochondrial locus. We discuss whether this group of embryos may represent an undescribed cryptic species. Although morphological description of adults is required for a complete species description, only embryo samples were available. Even so, results presented here are a step toward understanding, managing, and conserving the diversity of skate species in the North Pacific Ocean.
Improvement and Evaluation of Copper Oxidation Experimental Procedure for the Introduction of the Law of Definite Proportion
Authors:   Faculty of Education, Chiba University, Yamashita Shuichi , Kashiwaguma Yasuyuki ,   Faculty of Education, Chiba University, Hayashi Hideko ,   Faculty of Education, Chiba University, Pietzner Verena ,   School of Mathematics and Science, Carl von Ossietzky University
Published: June 2021

In Science classes, students usually learn about the law of definite proportions by the oxidation of copper. However, common procedures usually do not lead to proper results. This leads to confusion among the students because their experimental results do not fit to the theoretical values. Therefore, we invented a new procedure for this experiment to improve the introduction of the law of definite proportions. The new procedure can also be done by the students, it is done within 15 minutes and provides theoretical data written in the textbook. To evaluate this improved procedure, we let 8th year students (n = 133) do the copper oxidation experiment in the traditional and the improved way. The oxidation ratio doing the improved method was 98%, while the traditional way was only 55%. Student's individual feedback indicates that the new method might be able to improve the understanding of the law of definite proportions.
Assessment of College Students' Knowledge and Attitudes Towards Solid Waste Management in North Central Zone of Nigeria
Authors:   Federal College of Education Pankshin, Dung Mohammed Dauda , Mankilik Mangut ,   Department of Science and Technology Education, Faculty of Education, University of Jos, Ozoji Bernadette Ebele ,   Department of Science and Technology Education, Faculty of Education, University of Jos
Published: June 2021

This study focused on assessment of colleges of education students' knowledge and attitudes towards solid waste management in the North-Central zone of Nigeria. The cross-sectional survey design was adopted. A Students' Knowledge and Attitudes towards Solid Waste Management Questionnaire was used to collect data from 1,800 students. The findings indicated that the students had a low knowledge level of solid waste management but their attitudes towards it were positive. A significant relationship was found between students' knowledge and their attitudes towards solid waste management. Furthermore, gender was shown not to have any significant influence on knowledge and attitudes of students towards solid waste management. It was concluded that although students had low knowledge of solid waste management, their attitudes towards it were positive. It was recommended that environmental education issues, especially, solid waste management should be incorporated into colleges of education curricula and properly taught for in-depth knowledge acquisition.
Learning Science: Pūtaiao in indigenous schools: A review of the literature
Authors:   Victoria University of Wellington, Smallbone Catherine , Rofe Craig ,   Victoria University of Wellington, Moeed Azra ,   Victoria University of Wellington
Published: June 2021

This paper presents a literature review of theory and briefly presents insight from a case study. The literature review attempts to explain what Pūtaiao is, how it is being taught, and the learning of Pūtaiao. It also investigates the Pūtaiao curriculum and the challenges currently being faced. The literature covers students from early childhood, primary, and secondary school, and in this review, it is mainly limited to Māori immersion classrooms and schools. Further work is needed focusing on teacher education and professional development of Pūtaiao teachers. Pūtaiao as a subject is facing several challenges that may be combated through more specific interventions - rather than studies investigating Kura as a whole. Despite the challenges faced, the literature overall has a positive outlook in which Pūtaiao is enjoyed by students who see it relates to their own knowledge and culture. The case study took place in a Māori medium school that caters for year 9-13 students. Two teachers from a Māori medium school (Kura) who had no background in science and their year 9 and 10 classes voluntarily participated in this research. Through support to teachers in the form of upskilling themselves and learning along with the students, teachers gained confidence and began to teach Pūtaiao. Teachers willingly learn new vocabulary to pass it on to their students. In addition, among mainstream schools, many of the studies had positive impacts by raising awareness among teachers of how to teach Māori students, and how to incorporate some of the values that are fundamental to Pūtaiao.
Effects of Oral Liposomal Glutathione in Altering the Immune Responses Against Mycobacterium tuberculosis and the Mycobacterium bovis BCG Strain in Individuals With Type 2 Diabetes
Authors: To Kimberly , Cao Ruoqiong , Yegiazaryan Aram , Owens James , Nguyen Timothy , Sasaninia Kayvan , Vaughn Charles , Singh Mohkam , Truong Edward , Medina Albert , Avitia Edith , Villegas Jose , Pham Christal , Sathananthan Airani , Venketaraman Vishwanath
Published: June 2021

The World Health Organization (WHO) has identified type 2 diabetes (T2DM) as a neglected, important, and re-emerging risk factor for tuberculosis (TB), especially in low and middle-income countries where TB is endemic. In this clinical trial study, oral liposomal glutathione supplementation (L-GSH) or placebo was given to individuals with T2DM to investigate the therapeutic effects of L-GSH supplementation. We report that L-GSH supplementation for 3 months in people with T2DM was able to reduce the levels of oxidative stress in all blood components and prevent depletion of glutathione (GSH) in this population known to be GSH deficient. Additionally, L-GSH supplementation significantly reduced the burden of intracellular mycobacteria withinin vitrogranulomas generated from peripheral blood mononuclear cells (PBMCs) of T2DM subjects. L-GSH supplementation also increased the levels of Th1-associated cytokines, IFN-γ, TNF-α, and IL-2 and decreased levels of IL-6 and IL-10. In conclusion our studies indicate that oral L-GSH supplementation in individuals with T2DM for three months was able to maintain the levels of GSH, reduce oxidative stress, and diminish mycobacterial burden withinin vitrogenerated granulomas of diabetics. L-GSH supplementation for 3 months in diabetics was also able to modulate the levels of various cytokines.
Discovering and Approaching Mature Happiness: The Implementation of the CasMac Model in a University English Class
Authors: Chen Chih-Hong , Chang Shu-Mei , Wu Huei-Min
Published: June 2021

The purpose of this study was to explore the effects of integrating meaning-centered positive education (MCPE) and the second wave positive psychology (PP2.0) into a university English speaking class. The study adopted Wong’s CasMac model of PP2.0 and designed a series of English lessons which aimed to understand the meaning of life through the perspectives of PP2.0 and its focus on MCPE. The participants were 38 university students, with upper-intermediate English proficiency, enrolled in an English speaking class. They participated in the English program for 15 weeks and 2 h each week. The quantitative data was collected from survey of the CasMac Measure of Character and analyzed with the pairedt-test method, and the qualitative data analysis was collected from students’ weekly learning sheets and journals. The results show that the integration of MCPE and PP2.0 in a university English class is feasible to enhance students’ understanding of mature happiness through the CasMac model and to promote their meanings in life. According to the research findings, it is suggested that the CasMac model can be applied to other fields or other groups who need help to enhance life meaning and improve wellbeing. Particularly under the pandemic of COVID-19, there are people encountering traumas, losses, and sorrows and it is crucial to transform sufferings with the support of approaching mature happiness.
Corticosteroid Biosynthesis Revisited: No Direct Hydroxylation of Pregnenolone by Steroid 21-Hydroxylase
Authors: Loke Steffen , Stoll Anna , Machalz David , Botrè Francesco , Wolber Gerhard , Bureik Matthias , Parr Maria Kristina
Published: June 2021

Cytochrome P450s (CYPs) are an essential family of enzymes in the human body. They play a crucial role in metabolism, especially in human steroid biosynthesis. Reactions catalyzed by these enzymes are highly stereo- and regio-specific. Lack or severe malfunctions of CYPs can cause severe diseases and even shorten life. Hence, investigations on metabolic reactions and structural requirements of substrates are crucial to gain further knowledge on the relevance of different enzymes in the human body functions and the origin of diseases. One key enzyme in the biosynthesis of gluco- and mineralocorticoids is CYP21A2, also known as steroid 21-hydroxylase. To investigate the steric and regional requirements of substrates for this enzyme, we performed whole-cell biotransformation assays using a strain of fission yeast Schizosaccharomyces pombe recombinantly expressing CYP21A2. The progestogens progesterone, pregnenolone, and their 17α-hydroxy-derivatives were used as substrates. After incubation, samples were analyzed using gas chromatography coupled to mass spectrometry. For progesterone and 17α-hydroxyprogesterone, their corresponding 21-hydroxylated metabolites 11-deoxycorticosterone and 11-deoxycortisol were detected, while after incubation of pregnenolone and 17α-hydroxypregnenolone, no hydroxylated product was observed. Findings were confirmed with authentic reference material. Molecular docking experiments agree with these results and suggest that interaction between the 3-oxo group and arginine-234 of the enzyme is a strict requirement. The presented results demonstrate once more that the presence of an oxo-group in position 3 of the steroid is indispensable, while a 3-hydroxy group prevents hydroxylation in position C-21 by CYP21A2. This knowledge may be transferred to other CYP21A2 substrates and hence help to gain essential insights into steroid metabolism.
Schizophrenia Plays a Negative Role in the Pathological Development of Myocardial Infarction at Multiple Biological Levels
Authors: Yang Xiaorong , Chen Yao , Wang Huiyao , Fu Xia , Kural Kamil Can , Cao Hongbao , Li Ying
Published: June 2021

It has shown that schizophrenia (SCZ) is associated with a higher chance of myocardial infarction (MI) and increased mortality. However, the underlying mechanism is largely unknown. Here, we first constructed a literature-based genetic pathway linking SCZ and MI, and then we tested the expression levels of the genes involved in the pathway by a meta-analysis using nine gene expression datasets of MI. In addition, a literature-based data mining process was conducted to explore the connection between SCZ at different levels: small molecules, complex molecules, and functional classes. The genetic pathway revealed nine genes connecting SCZ and MI. Specifically, SCZ activates two promoters of MI (IL6 and CRP) and deactivates seven inhibitors of MI (ADIPOQ, SOD2, TXN, NGF, ADORA1, NOS1, and CTNNB1), suggesting that no protective role of SCZ in MI was detected. Meta-analysis showed that one promoter of MI (CRP) presented no significant increase, and six out of seven genetic inhibitors of MI demonstrated minor to moderately increased expression. Therefore, the elevation of CRP and inhibition of the six inhibitors of MI by SCZ could be critical pathways to promote MI. Nine other regulators of MI were influenced by SCZ, including two gene families (inflammatory cytokine and IL1 family), five small molecules (lipid peroxide, superoxide, ATP, ascorbic acid, melatonin, arachidonic acid), and two complexes (CaM kinase 2 and IL23). Our results suggested that SCZ promotes the development and progression of MI at different levels, including genes, small molecules, complex molecules, and functional classes.
A Scalable Strand-Specific Protocol Enabling Full-Length Total RNA Sequencing From Single Cells
Authors: Haile Simon , Corbett Richard D. , LeBlanc Veronique G. , Wei Lisa , Pleasance Stephen , Bilobram Steve , Nip Ka Ming , Brown Kirstin , Trinh Eva , Smith Jillian , Trinh Diane L. , Bala Miruna , Chuah Eric , Coope Robin J. N. , Moore Richard A. , Mungall Andrew J. , Mungall Karen L. , Zhao Yongjun , Hirst Martin , Aparicio Samuel , Birol Inanc , Jones Steven J. M. , Marra Marco A.
Published: June 2021

RNA sequencing (RNAseq) has been widely used to generate bulk gene expression measurements collected from pools of cells. Only relatively recently have single-cell RNAseq (scRNAseq) methods provided opportunities for gene expression analyses at the single-cell level, allowing researchers to study heterogeneous mixtures of cells at unprecedented resolution. Tumors tend to be composed of heterogeneous cellular mixtures and are frequently the subjects of such analyses. Extensive method developments have led to several protocols for scRNAseq but, owing to the small amounts of RNA in single cells, technical constraints have required compromises. For example, the majority of scRNAseq methods are limited to sequencing only the 3′ or 5′ termini of transcripts. Other protocols that facilitate full-length transcript profiling tend to capture only polyadenylated mRNAs and are generally limited to processing only 96 cells at a time. Here, we address these limitations and present a novel protocol that allows for the high-throughput sequencing of full-length, total RNA at single-cell resolution. We demonstrate that our method produced strand-specific sequencing data for both polyadenylated and non-polyadenylated transcripts, enabled the profiling of transcript regions beyond only transcript termini, and yielded data rich enough to allow identification of cell types from heterogeneous biological samples.
Bioinformatics Analysis of Differentially Expressed Rhythm Genes in Liver Hepatocellular Carcinoma
Authors: Liu Huaifeng , Gao Yu , Hu Shangshang , Fan Zhengran , Wang Xianggang , Li Shujing
Published: June 2021

Liver Hepatocellular Carcinoma (LIHC), a malignant tumor with high incidence and mortality, is one of the most common cancers in the world. Multiple studies have found that the aberrant expression of rhythm genes is closely related to the occurrence of LIHC. This study aimed to use bioinformatics analysis to identify differentially expressed rhythm genes (DERGs) in LIHC. A total of 563 DERGs were found in LIHC, including 265 downregulated genes and 298 upregulated genes. KEGG pathway enrichment and GO analyses showed that DERGs were significantly enriched in rhythmic and metabolic processes. Survival analysis revealed that high expression levels of CNK1D, CSNK1E, and NPAS2 were significantly associated with the low survival rate in LIHC patients. Through cell experiment verification, the mRNA expression levels of CSNK1D, CSNK1E, and NPAS2 were found to be strongly upregulated, which was consistent with the bioinformatics analysis of LIHC patient samples. A total of 23 nodes and 135 edges were involved in the protein–protein interaction network of CSNK1D, CSNK1E, and NPAS2 genes. Clinical correlation analyses revealed that CSNK1D, CSNK1E, and NPAS2 expression levels were high-risk factors and independently connected with the overall survival rate in LIHC patients. In conclusion, the identification of these DERGs contributes to the exploration of the molecular mechanisms of LIHC occurrence and development and may be used as diagnostic and prognostic biomarkers and molecular targets for chronotherapy in LIHC patients in the future.
Modulation of Intestinal ILC3 for the Treatment of Type 1 Diabetes
Authors: Stojanović Ivana , Saksida Tamara , Miljković Đorđe , Pejnović Nada
Published: June 2021

Gut-associated lymphoid tissue (GALT) is crucial for the maintenance of the intestinal homeostasis, but it is also the potential site of the activation of autoreactive cells and initiation/propagation of autoimmune diseases in the gut and in the distant organs. Type 3 innate lymphoid cells (ILC3) residing in the GALT integrate signals from food ingredients and gut microbiota metabolites in order to control local immunoreactivity. Notably, ILC3 secrete IL-17 and GM-CSF that activate immune cells in combating potentially pathogenic microorganisms. ILC3 also produce IL-22 that potentiates the strength and integrity of epithelial tight junctions, production of mucus and antimicrobial peptides thus enabling the proper function of the intestinal barrier. The newly discovered function of small intestine ILC3 is the secretion of IL-2 and the promotion of regulatory T cell (Treg) generation and function. Since the intestinal barrier dysfunction, together with the reduction in small intestine ILC3 and Treg numbers are associated with the pathogenesis of type 1 diabetes (T1D), the focus of this article is intestinal ILC3 modulation for the therapy of T1D. Of particular interest is free fatty acids receptor 2 (FFAR2), predominantly expressed on intestinal ILC3, that can be stimulated by available selective synthetic agonists. Thus, we propose that FFAR2-based interventions by boosting ILC3 beneficial functions may attenuate autoimmune response against pancreatic β cells during T1D. Also, it is our opinion that treatments based on ILC3 stimulation by functional foods can be used as prophylaxis in individuals that are genetically predisposed to develop T1D.
Identification and Characterization of the Amphioxus Lck and Its Associated Tyrosine Phosphorylation-Dependent Inhibitory LRR Receptor
Authors: Zhou Jiatao , Xiao Zhihui , Zhan Yanli , Qu Xuemei , Mou Sisi , Deng Chong , Zhang Tianxiang , Lan Xin , Huang Shengfeng , Li Yingqiu
Published: June 2021

Amphioxus (e.g.,Branchiostoma belcheri, Bb) has recently emerged as a new model for studying the origin and evolution of vertebrate immunity. Mammalian lymphocyte-specific tyrosine kinase (Lck) plays crucial roles in T cell activation, differentiation and homeostasis, and is reported to phosphorylate both the ITIM and ITSM of PD-1 to induce the recruitment of phosphatases and thus the inhibitory function of PD-1. Here, we identified and cloned the amphioxus homolog of human Lck. By generating and using an antibody against BbLck, we found that BbLck is expressed in the amphioxus gut and gill. Through overexpression of BbLck in Jurkat T cells, we found that upon TCR stimulation, BbLck was subjected to tyrosine phosphorylation and could partially rescue Lck-dependent tyrosine phosphorylation in Lck-knockdown T cells. Mass spectrometric analysis of BbLck immunoprecipitates from immunostimulants-treated amphioxus, revealed a BbLck-associated membrane-bound receptor LRR (BbLcLRR). By overexpressing BbLcLRR in Jurkat T cells, we demonstrated that BbLcLRR was tyrosine phosphorylated upon TCR stimulation, which was inhibited by Lck knockdown and was rescued by overexpression of BbLck. By mutating single tyrosine to phenylalanine (Y-F), we identified three tyrosine residues (Y539, Y655, and Y690) (3Y) of BbLcLRR as the major Lck phosphorylation sites. Reporter gene assays showed that overexpression of BbLcLRR but not the BbLcLRR-3YF mutant inhibited TCR-induced NF-κB activation. In Lck-knockdown T cells, the decline of TCR-induced IL-2 production was reversed by overexpression of BbLck, and this reversion was inhibited by co-expression of BbLcLRR but not the BbLcLRR-3YF mutant. Sequence analysis showed that the three tyrosine-containing sequences were conserved with the tyrosine-based inhibition motifs (ITIMs) or ITIM-like motifs. And TCR stimulation induced the association of BbLcLRR with tyrosine phosphatases SHIP1 and to a lesser extent with SHP1/2. Moreover, overexpression of wild-type BbLcLRR but not its 3YF mutant inhibited TCR-induced tyrosine phosphorylation of multiple signaling proteins probablyviarecruiting SHIP1. Thus, we identified a novel immunoreceptor BbLcLRR, which is phosphorylated by Lck and then exerts a phosphorylation-dependent inhibitory role in TCR-mediated T-cell activation, implying a mechanism for the maintenance of self-tolerance and homeostasis of amphioxus immune system and the evolutionary conservatism of Lck-regulated inhibitory receptor pathway.
Optical Control of CD8+ T Cell Metabolism and Effector Functions
Authors: Amitrano Andrea M. , Berry Brandon J. , Lim Kihong , Kim Kyun-Do , Waugh Richard E. , Wojtovich Andrew P. , Kim Minsoo
Published: June 2021

Although cancer immunotherapy is effective against hematological malignancies, it is less effective against solid tumors due in part to significant metabolic challenges present in the tumor microenvironment (TME), where infiltrated CD8+ T cells face fierce competition with cancer cells for limited nutrients. Strong metabolic suppression in the TME is often associated with impaired T cell recruitment to the tumor site and hyporesponsive effector function via T cell exhaustion. Increasing evidence suggests that mitochondria play a key role in CD8+ T cell activation, effector function, and persistence in tumors. In this study, we showed that there was an increase in overall mitochondrial function, including mitochondrial mass and membrane potential, during both mouse and human CD8+ T cell activation. CD8+ T cell mitochondrial membrane potential was closely correlated with granzyme B and IFN-γ production, demonstrating the significance of mitochondria in effector T cell function. Additionally, activated CD8+ T cells that migrate on ICAM-1 and CXCL12 consumed significantly more oxygen than stationary CD8+ T cells. Inhibition of mitochondrial respiration decreased the velocity of CD8+ T cell migration, indicating the importance of mitochondrial metabolism in CD8+ T cell migration. Remote optical stimulation of CD8+ T cells that express our newly developed “OptoMito-On” successfully enhanced mitochondrial ATP production and improved overall CD8+ T cell migration and effector function. Our study provides new insight into the effect of the mitochondrial membrane potential on CD8+ T cell effector function and demonstrates the development of a novel optogenetic technique to remotely control T cell metabolism and effector function at the target tumor site with outstanding specificity and temporospatial resolution.
Tuberculosis Risk Stratification of Psoriatic Patients Before Anti-TNF-α Treatment
Authors: Benhadou Farida , Dirix Violette , Domont Fanny , Willaert Fabienne , Van Praet Anne , Locht Camille , Mascart Françoise , Corbière Véronique
Published: June 2021

Psoriasis is a skin inflammatory condition for which significant progress has been made in its management by the use of targeted biological drugs. Detection of latent M. tuberculosis infection (LTBI) is mandatory before starting biotherapy that is associated with reactivation risk. Together with evaluation of TB risk factors and chest radiographs, tuberculin skin tests (TST) and/or blood interferon-γ-release assays (IGRA), like the QuantiFERON (QFT), are usually performed to diagnose M. tuberculosis infection. Using this approach, 14/49 psoriatic patients prospectively included in this study were identified as LTBI (14 TST+, induration size ≥ 10mm, 8 QFT+), and 7/14 received prophylactic anti-TB treatment, the other 7 reporting past-treatment. As the specificity and sensitivity of these tests were challenged, we evaluated the added value of an IGRA in response to a mycobacterial antigen associated with latency, the heparin-binding haemagglutinin (HBHA). All but one TST+ patient had a positive HBHA-IGRA, indicating higher sensitivity than the QFT. The HBHA-IGRA was also positive for 12/35 TST-QFT- patients. Measurement for 15 psoriatic patients (12 with HBHA-IGRA+) of 8 chemokines in addition to IFN-γ revealed a broad array of HBHA-induced chemokines for TST+QFT- and TST-QFT- patients, compared to a more restricted pattern for TST+QFT+ patients. This allowed us to define subgroups within psoriatic patients characterized by different immune responses to M. tuberculosis antigens that may be associated to different risk levels of reactivation of the infection. This approach may help in prioritizing patients who should receive prophylactic anti-TB treatment before starting biotherapies in order to reduce their number.
Inefficient Placental Virus Replication and Absence of Neonatal Cell-Specific Immunity Upon Sars-CoV-2 Infection During Pregnancy
Authors: Tallarek Ann-Christin , Urbschat Christopher , Fonseca Brito Luis , Stanelle-Bertram Stephanie , Krasemann Susanne , Frascaroli Giada , Thiele Kristin , Wieczorek Agnes , Felber Nadine , Lütgehetmann Marc , Markert Udo R. , Hecher Kurt , Brune Wolfram , Stahl Felix , Gabriel Gülsah , Diemert Anke , Arck Petra Clara
Published: June 2021

Pregnant women have been carefully observed during the COVID-19 pandemic, as the pregnancy-specific immune adaptation is known to increase the risk for infections. Recent evidence indicates that even though most pregnant have a mild or asymptomatic course, a severe course of COVID-19 and a higher risk of progression to diseases have also been described, along with a heightened risk for pregnancy complications. Yet, vertical transmission of the virus is rare and the possibility of placental SARS-CoV-2 infection as a prerequisite for vertical transmission requires further studies. We here assessed the severity of COVID-19 and onset of neonatal infections in an observational study of women infected with SARS-CoV-2 during pregnancy. Our placental analyses showed a paucity of SARS-CoV-2 viral expression ex vivo in term placentae under acute infection. No viral placental expression was detectable in convalescent pregnant women. Inoculation of placental explants generated from placentas of non-infected women at birth with SARS-CoV-2 in vitro revealed inefficient SARS-CoV-2 replication in different types of placental tissues, which provides a rationale for the low ex vivo viral expression. We further detected specific SARS-CoV-2 T cell responses in pregnant women within a few days upon infection, which was undetectable in cord blood. Our present findings confirm that vertical transmission of SARS-CoV-2 is rare, likely due to the inefficient virus replication in placental tissues. Despite the predominantly benign course of infection in most mothers and negligible risk of vertical transmission, continuous vigilance on the consequences of COVID-19 during pregnancy is required, since the maternal immune activation in response to the SARS-CoV2 infection may have long-term consequences for children’s health.
Neuronal Damage and Neuroinflammation, a Bridge Between Bacterial Meningitis and Neurodegenerative Diseases
Authors: Farmen Kristine , Tofiño-Vian Miguel , Iovino Federico
Published: June 2021

Bacterial meningitis is an inflammation of the meninges which covers and protects the brain and the spinal cord. Such inflammation is mostly caused by blood-borne bacteria that cross the blood-brain barrier (BBB) and finally invade the brain parenchyma. Pathogens such asStreptococcus pneumoniae,Neisseria meningitidis, andHaemophilus influenzaeare the main etiological causes of bacterial meningitis. After trafficking across the BBB, bacterial pathogens in the brain interact with neurons, the fundamental units of Central Nervous System, and other types of glial cells. Although the specific molecular mechanism behind the interaction between such pathogens with neurons is still under investigation, it is clear that bacterial interaction with neurons and neuroinflammatory responses within the brain leads to neuronal cell death. Furthermore, clinical studies have shown indications of meningitis-caused dementia; and a variety of neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease and Huntington’s disease are characterized by the loss of neurons, which, unlike many other eukaryotic cells, once dead or damaged, they are seldom replaced. The aim of this review article is to provide an overview of the knowledge on how bacterial pathogens in the brain damage neurons through direct and indirect interactions, and how the neuronal damage caused by bacterial pathogen can, in the long-term, influence the onset of neurodegenerative disorders.