Comparative analysis of SARS-CoV-2 variants Alpha (B.1.1.7), Gamma (P.1), Zeta (P.2) and Delta (B.1.617.2) in Vero-E6 cells: ultrastructural characterization of cytopathology and replication kinetics.

This study compares the effects of virus-cell interactions among SARS-CoV-2 variants of concern (VOCs) isolated in Brazil in 2021, hypothesizing a correlation between cellular alterations and mortality and between viral load and transmissibility. For this purpose, reference isolates of Alpha, Gamma, Zeta, and Delta variants were inoculated into monolayers of Vero-E6 cells. Viral RNA was quantified in cell supernatants by RT‒PCR, and infected cells were analyzed by Transmission Electron Microscopy (TEM) for qualitative and quantitative evaluation of cellular changes 24, 48, and 72 hours postinfection (hpi). Ultrastructural analyses showed that all variants of SARS-CoV-2 altered the structure and function of mitochondria, nucleus, and rough endoplasmic reticulum of cells. Monolayers infected with the Delta variant showed the highest number of modified cells and the greatest statistically significant differences compared to those of other variants. Viral particles were observed in the cytosol and the cell membrane in 100 % of the cells at 48 hpi. Alpha showed the highest mean particle diameter (79 nm), and Gamma and Delta were the smallest (75 nm). Alpha and Gamma had the highest particle frequency per field at 48 hpi, while the same was observed for Zeta and Delta at 72 hpi and 24 hpi, respectively. The cycle threshold of viral RNA varied among the target protein, VOC, and time of infection. The findings presented here demonstrate that all four VOCs evaluated caused ultrastructural changes in Vero-E6 cells, which were more prominent when infection occured with the Delta variant.

SARS-CoV-2 intra-host diversity, antibody response, and disease severity after reinfection by the variant of concern Gamma in Brazil.

The rapid spread of the SARS-CoV-2 Variant of Concern (VOC) Gamma in Amazonas during early 2021 fueled a second large COVID-19 epidemic wave and raised concern about the potential role of reinfections. Very few cases of reinfection associated with the VOC Gamma have been reported to date, and their potential impact on clinical, immunological, and virological parameters remains largely unexplored. Here we describe 25 cases of SARS-CoV-2 reinfection in Brazil. SARS-CoV-2 genomic analysis confirmed that individuals were primo-infected with distinct viral lineages between March and December 2020 (B.1.1, B.1.1.28, B.1.1.33, B.1.195, and P.2) and reinfected with the VOC Gamma between 3 to 12 months after primo-infection. We found a similar mean cycle threshold (Ct) value and limited intra-host viral diversity in both primo-infection and reinfection samples. Sera of 14 patients tested 10-75 days after reinfection displayed detectable neutralizing antibodies (NAb) titers against SARS-CoV-2 variants that circulated before (B.1.*), during (Gamma), and after (Delta and Omicron) the second epidemic wave in Brazil. All individuals had milder or no symptoms after reinfection, and none required hospitalization. These findings demonstrate that individuals reinfected with the VOC Gamma may display relatively high RNA viral loads at the upper respiratory tract after reinfection, thus contributing to onward viral transmissions. Despite this, our study points to a low overall risk of severe Gamma reinfections, supporting that the abrupt increase in hospital admissions and deaths observed in Amazonas and other Brazilian states during the Gamma wave was mostly driven by primary infections. Our findings also indicate that most individuals analyzed developed a high anti-SARS-CoV-2 NAb response after reinfection that may provide some protection against reinfection or disease by different SARS-CoV-2 variants.

Sera of patients infected by earlier lineages of SARS-CoV-2 are capable to neutralize later emerged variants of concern.

Serum samples of 20 hospitalized coronavirus disease 2019 (COVID-19) patients from Brazil who were infected by the earlier severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineages B.1.1.28 and B.1.1.33, and by the variant of concern (VOC) Gamma (P.1) were tested by plaque reduction neutralization test (PRNT90) with wild isolates of a panel of SARS-CoV-2 lineages, including B.1, Zeta, N.10, and the VOCs Gamma, Alpha, and Delta that emerged in different timeframes of the pandemic. The main objective of this study was to evaluate if the serum of patients infected by earlier lineages was capable to neutralize later emerged VOCs. We also evaluated if the 4-fold difference in PRNT90 titers is a reliable seropositivity criterion to distinguish infections caused by different SARS-CoV-2 lineages. Sera collected between May 2020 and August 2021 from the day of admittance to the hospital to 21 days after diagnostic of patients infected by the two earlier lineages B.1.1.28 and B.1.1.33 presented neutralizing capacity for all challenged VOCs, including Gamma and Delta. Among all variants tested, Delta and N.10 presented the lowest geometric mean of neutralizing antibody titers, and B.1.1.7, presented the highest titers. Four patients infected with Gamma, that emerged in December 2020, presented neutralizing antibodies for B.1, B.1.1.33, and B.1.1.28, its ancestor lineage. All of them had neutralizing antibodies under the level of detection for the VOC Delta. Patients infected by B.1.1.28 presented very similar geometric mean of neutralizing antibody titers for both B.1.1.33 and B.1.1.28. Findings presented here indicate that most patients infected in early stages of COVID-19 pandemic presented neutralizing antibodies capable to neutralize wild types of all later emerged VOCs in Brazil, and that the 4-fold difference in PRNT90 titers is not reliable to distinguish humoral response among different SARS-CoV-2 lineages.

Macrophage Resistance to Ionizing Radiation Exposure Is Accompanied by Decreased Cathepsin D and Increased Transferrin Receptor 1 Expression.

PURPOSE: To identify a molecular signature of macrophages exposed to clinically relevant ionizing radiation (IR) doses, mirroring radiotherapy sessions. METHODS: Human monocyte-derived macrophages were exposed to 2 Gy/ fraction/ day for 5 days, mimicking one week of cancer patient's radiotherapy. Protein expression profile by proteomics was performed. RESULTS: A gene ontology analysis revealed that radiation-induced protein changes are associated with metabolic alterations, which were further supported by a reduction of both cellular ATP levels and glucose uptake. Most of the radiation-induced deregulated targets exhibited a decreased expression, as was the case of cathepsin D, a lysosomal protease associated with cell death, which was validated by Western blot. We also found that irradiated macrophages exhibited an increased expression of the transferrin receptor 1 (TfR1), which is responsible for the uptake of transferrin-bound iron. TfR1 upregulation was also found in tumor-associated mouse macrophages upon tumor irradiation. In vitro irradiated macrophages also presented a trend for increased divalent metal transporter 1 (DMT1), which transports iron from the endosome to the cytosol, and a significant increase in iron release. CONCLUSIONS: Irradiated macrophages present lower ATP levels and glucose uptake, and exhibit decreased cathepsin D expression, while increasing TfR1 expression and altering iron metabolism.

Comparative analysis of SARS-CoV-2 variants Alpha (B.1.1.7), Gamma (P.1), Zeta (P.2) and Delta (B.1.617.2) in Vero-E6 cells: ultrastructural characterization of cytopathology and replication kinetics

Abstract This study compares the effects of virus-cell interactions among SARS-CoV-2 variants of concern (VOCs) isolated in Brazil in 2021, hypothesizing a correlation between cellular alterations and mortality and between viral load and transmissibility. For this purpose, reference isolates of Alpha, Gamma, Zeta, and Delta variants were inoculated into monolayers of Vero-E6 cells. Viral RNA was quantified in cell supernatants by RT‒PCR, and infected cells were analyzed by Transmission Electron Microscopy (TEM) for qualitative and quantitative evaluation of cellular changes 24, 48, and 72 hours postinfection (hpi). Ultrastructural analyses showed that all variants of SARS-CoV-2 altered the structure and function of mitochondria, nucleus, and rough endoplasmic reticulum of cells. Monolayers infected with the Delta variant showed the highest number of modified cells and the greatest statistically significant differences compared to those of other variants. Viral particles were observed in the cytosol and the cell membrane in 100 % of the cells at 48 hpi. Alpha showed the highest mean particle diameter (79 nm), and Gamma and Delta were the smallest (75 nm). Alpha and Gamma had the highest particle frequency per field at 48 hpi, while the same was observed for Zeta and Delta at 72 hpi and 24 hpi, respectively. The cycle threshold of viral RNA varied among the target protein, VOC, and time of infection. The findings presented here demonstrate that all four VOCs evaluated caused ultrastructural changes in Vero-E6 cells, which were more prominent when infection occured with the Delta variant.

Educação interprofissional: um relato de experiência para promoção de uma alimentação adequada e saudável / Interprofessional education: a report of experience to promote an appropriate and healthy food / Educación interprofesional: un informe de experiencia para promover una alimentación apropiada y saludable

Introdução:A educação interprofissional vem se destacando Brasil devido ao reconhecimento da capacidade da abordagem em melhorar a qualidade dos serviços de saúde, comunicação e interação entre estudantes.Objetivo:Relatar uma experiência de acadêmicos doscursosde medicina e nutrição em uma atividadeeducativa interprofissionalem saúdepara promoção de uma alimentação adequada e saudável, para crianças em idade pré-escolar, realizada no segundo semestre do ano de 2019, em uma escola municipal,Anápolis,Goiás.Metodologia:Trata-se de um estudo descritivo, tipo relato de experiência, elaborado por estudantes da área da saúde após projeto realizado com crianças empré-escolaridade escolar. Resultados:A atividade educativa foi idealizada após análise referente às doenças crônicas do território da Unidade Básica de Saúde assistida em conjunto com a disciplina Medicina de Família e Comunidade e verificou-se a necessidade de trabalhar a conscientização acerca da alimentação com as crianças pré-escolarpor ser uma idade de descoberta e formação dos hábitos alimentares. Por meio da parceria entre os dois cursos, foram elaboradas atividades lúdicas e a diferenciação dos alimentos para fomentar a criação de hábitos alimentares saudáveis e assim evitar e/ou minimizar possíveis patologias no futuro. Conclusões:Conclui-se que a educação em saúde aliada a interprofissionalidadeé de suma importância para o aprendizado, tendo sido proporcionado um momento de troca de conhecimentos, tanto entre os acadêmicosde diferentes cursos e professores, quanto entre acadêmicos e crianças (AU).

Introduction:Interprofessional education has been standing out in Brazil due to the recognition of the approach's ability to improve the quality of health services, communication and interaction among students. Objective:To report an experience of medical and nutrition students in an interprofessional health education activity to promote adequate and healthy eating for preschool-age children, held in the second half of 2019, in a municipal school, Anápolis, Goiás.Methodology:This is a descriptive study, type of experience report, prepared by students in the health area after a project carried out with school-aged children of 4years. Results:The action was idealized after analysis regarding chronic diseases in the basic health Unitterritory'sassisted in conjunction with the Family and Community Medicine discipline. Therefore, there was a need to raise awareness about eating with 4-year-old children from a municipal school, as it is an age of discovery and formation of eating habits. Through the partnership between the medical course and the nutrition course, recreational activities were developed, in addition to the recognition of foods and the differentiation of fresh and processed foods, to promote the creation of healthy eating habits and thus avoid and/or minimize possible pathologies in the future. Conclusions:Under this point of view, it is concluded that Health Education combined with interdisciplinarity is extremely important for learning, having provided a moment of exchange of knowledge, both among students from different courses and teachers, as well as between academics and children (AU).

Introducción: La educación interprofesionalse ha destacado en Brasil por el reconocimiento de la capacidad del enfoque para mejorar la calidad de los servicios de salud, la comunicación y la interacción entre los estudiantes.Objetivo: Informar una experiencia de estudiantes de medicina y nutrición en una actividad interprofesional de educación en salud para promover una alimentación adecuada y saludable para niños en edad preescolar, realizada en el segundo semestre de 2019, en una escuela municipal, Anápolis, Goiás.Metodología: Se trata de un estudio descriptivo, tipo de relato de experiencia, elaborado por estudiantes del ámbito de la salud a partir de un proyecto realizado con niños en edad escolar de 4 años.Resultados:La acción fue concebida luego de un análisis sobre enfermedades crónicas en el territoriode La Unidad básica de salud asistidaen conjunto con la disciplina de Medicina Familiar y Comunitaria y existía la necesidad de sensibilizar sobre la alimentación a los niños de 4 años ya que es una época de descubrimiento y formación de hábitos alimenticios. A través de la alianza entre los dos cursos, se desarrollaron actividades lúdicas y diferenciación alimentaria para fomentar la creación de hábitos alimentarios saludables y así evitar y / o minimizar posibles patologías en el futuro.Conclusiones: Se concluye que la Educación para la Salud combinada con la interdisciplinariedad es de suma importancia para el aprendizaje, habiendo brindado un momento de intercambio de conocimientos, tanto entre estudiantes de diferentes cursos y docentes, como entre académicos y niños (AU).

Fatores determinantes da restrição do crescimento extra-uterino em recém nascidos de muito baixo peso no período intra-hospitalar

Analisa o efeito das morbidades neonatais, práticas clínicas, características demográficas e das variáveis que refletem as práticas nutricionais no escore z do peso e perímetro cefálico (PC) para idade gestacional corrigida na alta hospitalar em recém-nascidos de muito baixo peso ao nascer.

Fatores determinantes da restrição do crescimento extra-uterino em Recém Nascidos de Muito Baixo Peso no período intra-hospitalar / Determinants of growth restriction extrauterine in Newborns of Very Low Birth Weight in the intra-hospital period

Os fatores determinantes do crescimento e do desenvolvimento adequados de Recém Nascidos (RN) prematuros ainda são pouco conhecidos. Apesar de o avanço tecnológico ter possibilitado sobrevivência cada vez maior de RN com peso de nascimento cada vez menor nas últimas décadas, seu crescimento continua consideravelmente insuficiente, mantendo-se assim até a vida adulta.1A nutrição e o monitoramento do ganho de peso dos RN de muito baixo peso permanecem como desafios aos neonatologistas. Adequar a melhor forma de nutrir esses recém-nascidos às práticas clínicas necessárias ao tratamento de morbidades inerentes à prematuridade, minimizar a perda de peso que ocorre nas duas primeiras semanas de vida, proporcionar oferta calórica suficiente tanto para uma menor restrição ao ganho ponderal quanto para assegurar seu crescimento, monitorar o ganho de peso usando curvas de avaliação e como incorporar novos conhecimentos às rotinas já estabelecidas são algumas questões abordadas por pesquisadores da área de nutrição para recém-nascidos prematuros de baixo peso ao nascer. Este trabalho tem como objetivo a análise do efeito das morbidades neonatais, práticas clínicas, características demográficas e das variáveis que refletem as práticas nutricionais no escore z do peso e perímetro cefálico (PC) para idade gestacional corrigida na alta hospitalar em recém-nascidos de muito baixo peso ao nascer.

The determinants of growth and development of appropriate newborn (NB) premature infants are still largely unknown. Although technological advances have enabled increasing survival of infants with birth weights less and less in recent decades, its growth remains insufficient, thus remaining until adulthood. Nutrition monitoring and weight gain of very low birth weight infants remain as challenges to neonatologists. Matching the best way to nurture these infants to clinical practice needed for treatment of morbidities associated with prematurity, minimize weight loss that occurs in the first two weeks of life, provide enough energy intake to a much less restriction as to weight gain ensure its growth, monitor weight gain using curves of evaluation and how to incorporate new knowledge to established routines are some issues raised by researchers in the field of nutrition for premature infants of low birth weight. This study aims to analyze the effect of neonatal morbidities, clinical practices, demographic characteristics and variables that reflect nutritional practices z scores for weight and head circumference (HC) corrected for gestational age at hospital discharge in infants of very low birth weight.