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.

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.

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.

Internação e alta hospitalar do recém-nascido na unidade de cuidado neonatal: identificação das dúvidas dos pais / Hospitalization and discharge of the neonate in the neonatal care unit: identification of parents' doubts / Hospitalización y alta del recién nacido en la unidad de cuidado neonatal: identificación de las dudas de los padres

Objetivo:Identificar as dúvidas dos pais frente aos cuidados dos seus filhos na UTIN no processo de internação e alta hospitalar. Metodologia: estudo de natureza descritiva e abordagem qualitativa. O cenário da pesquisa foi uma unidade neonatal de um hospital terciário, localizado no Estado do Rio de Janeiro. Foram selecionadas, por conveniência, como participantes do estudo as mães e pais dos recém-nascidos internados na UTIN, com internação mínima de 2 semanas, e que estivessem presentes na unidade. A coleta de dados foi realizada através de dados obtidos no prontuário do recém-nascido e um questionário. Foram realizadas também entrevistas abertas com os pais na primeira semana de internação do RN na UTIN e nas semanas subsequentes, até a semana da alta. Nesta pesquisa, a análise de dados foi feita por meio da análise de conteúdo de Bardin. Resultados: Participaram desta pesquisa 10 mães e 02 pais de RNinternados na UTIN. Com a transcrição das entrevistas, emergiram setecategorias que se tratavam das dúvidas que as mães e os pais apresentavam durante o processo de internação, descritas a seguir: Cuidados Básicos; Amamentação/Alimentação; Tratamento; Exames/Cirurgias; Impeditivos para alta; Cuidados pós-cirúrgicos; Tempo de internação. A categoria com o maior número de dúvidas apresentada durante a pesquisa foi sobre Tratamento (25.5%). Conclusão:Foi possível constatarque a internação na UTIN gera nos pais diversas dúvidas. Dúvidas essas relacionadas aos cuidados prestados aos seus filhos, sobre amamentação e alimentação, sobre o tratamento, entre outras.

Objective: To identify the doubts of parents regarding the care of their babies in the NICU in the process of hospitalization and hospital discharge. Methodology: descriptive study and qualitative approach. The research scenario was a neonatal unit of a tertiary hospital, located in the Rio de Janeiro, Brazil. The mothers and fathers of newborns admitted to the NICU, with a minimum stay of 2 weeks, and who were present in the unit, were selected, for convenience, as study participants. Data collection was performed using data obtained from the newborn's medical record and a questionnaire. Open interviews were also carried out with the parents in the first week of the newborn's hospitalization in the NICU and in the subsequent weeks, until the week of discharge. In this research, data analysis was performed using Bardin's content analysis. Results: 10 mothers and 02 fathers of NBadmitted to the NICU participated in this research. With the transcription of the interviews, seven categories emerged that dealt with the doubts that mothers and fathers had during the hospitalization process, described below: Basic Care; Breastfeeding/Feeding; Treatment; Exams/Surgeries; Disadvantages for discharge; Post-surgical care; Hospitalization time. The category with the highest number of doubts presented during the survey was Treatment (25.5%). Conclusion: It was possible to verify that the hospitalization in the NICU generates several doubts in the parents. Doubts related to the care provided to their children, about breastfeeding and feeding, about treatment, among others.

Objetivo: Identificar las dudas de los padres sobre el cuidado de sus hijos en la UCIN en proceso de hospitalización y alta hospitalaria. Metodología: estudio descriptivo y enfoque cualitativo. El escenario de la investigación fue una unidad neonatal de un hospital de tercer nivel, ubicado en el Estado de Río de Janeiro. Las madres y padres de recién nacidos ingresados en la UCIN, con estancia mínima de 2 semanas, y que estuvieran presentes en la unidad, fueron seleccionados, por conveniencia, como participantes del estudio. La recolección de datos se realizó a partir de los datos obtenidos de la historia clínica del recién nacido y de un cuestionario. También se realizaron entrevistas abiertas con los padres en la primera semana de internación del RN en la UCIN y en las semanas posteriores, hasta la semana del alta. En esta investigación, el análisis de datos se realizó mediante el análisis de contenido de Bardin. Resultados: Participaron de esta investigación 10 madres y 02 padres de RN ingresados en la UCIN. Con la transcripción de las entrevistas, surgieron siete categorías que abordaban las dudas que las madres y los padres tenían durante el proceso de hospitalización, descritas a continuación: Atención Básica; Lactancia materna/Alimentación; Tratamiento; Exámenes/Cirugías; Desventajas para la descarga; Atención posquirúrgica; Tiempo de hospitalización. La categoría con mayor número de dudas presentadas durante la encuesta fue Tratamiento (25,5%). Conclusión: Se pudo verificar que la internación en la UCIN genera varias dudas en los padres. Dudas relacionadas con la atención brindada a sus hijos, sobre la lactancia y alimentación, sobre el tratamiento, entre otros.

Bem-vindo ao lar: dificuldades dos cuidadores de bebês nascidos prematuramente após a alta hospitalar / Welcome home: difficulties of caregivers of premature infants after hospital discharge / Bienvenido a casa: dificultades de los cuidadores de bebés prematuros después del alta del hospital

Objetivo: Identificar as dificuldades dos cuidadores após a alta hospitalar da UTI Neonatal. Metodologia: Pesquisa de abordagem quantitativa, tipo transversal de caráter descritivo e exploratório. Desenvolvido no ambulatório de seguimento/ Follow-up de um hospital terciário no Rio de Janeiro.Os voluntáriosda pesquisa foram os pais ou cuidadores de recém-nascidos com idade gestacional menor que 37 semanas e que tiveram o tempo mínimo de quinze dias de internação do filho na UTIN. Os dados foram coletados através de uma entrevista na primeira consulta do RN no follow-up, realizada após uma semana de alta. A análise dos dados foi feita por meio deestatística descritiva. Resultados: Foram incluídos no estudo 15 cuidadores. Dentre as orientações que não foram recebidas destaca-se os sinais de alerta (70,6%), Limpeza da mamadeira (52,9%) e uso da chupeta (52,9%). Ao relacionar o tempo de permanência na UTIN com as orientações recebidas vemos que os cuidadores em que o RN permanece por menor tempo na UTIN são os que acabam tendo menos orientações sobre os cuidados com atemperatura, cuidados com higiene e sinais de alerta. Conclusão: Os resultados aqui apresentados mostraram que a chegada da família ao domicílio com o bebê, representa orompimento com o mundo da internação e gera experiências próprias do contexto domiciliar.

Objective: To identify the difficulties of caregivers after hospital discharge from the Neonatal ICU. Methodology: Research with a quantitative approach, transversal type of descriptive and exploratory character. Developed in the follow-up/Follow-up clinic of a tertiary hospital in Rio de Janeiro. The research volunteers were parents or caregivers of preterm infants with a gestational age of less than 37 weeks and who had at least fifteen days of hospitalization. Data were collected through an interview during first follow-up appointment, carried out after one week of discharge. Data analysis was performed using descriptive statistics. Results: 15 caregivers were included in the study. Among the guidelines that were not received, warning signs (70.6%), bottle cleaning (52.9%) and pacifier use (52.9%) stand out. When relating the length of stay in the NICU with the guidelines received, we see that caregivers whose newborns spend less time in the NICU end up having less guidance on temperature care, hygiene care and warning signs. Conclusion: The results presented here showed that the arrival of the family at home with the baby represents a break with the world of hospitalization and generates experiences specific to the home context.

Objetivo: Identificar las dificultades de los cuidadores después del alta hospitalaria de la UTI Neonatal.Metodología: Investigación con enfoque cuantitativo, de tipo transversal de carácter descriptivo y exploratorio. Desarrollado en el ambulatorio de seguimiento/seguimiento de un hospital de tercer nivel en Río de Janeiro. Los voluntarios de la investigación fueron los padres o cuidadores de recién nacidos con edad gestacional menor de 37 semanas y que tuvieran un tiempo mínimo de quince días de internación del niño en la UCIN. Los datos fueron recolectados a través de una entrevista en la primera visita de seguimiento del RN, realizada después de una semana del alta. El análisis de los datos se realizó mediante estadística descriptiva. Resultados: Quince cuidadores fueron incluidos en el estudio. Entre las orientaciones que no fueron recibidas, se destacan las señales de advertencia (70,6%), limpieza del biberón (52,9%) y uso del chupete (52,9%). Al relacionar el tiempo de permanencia en la UCIN con las orientaciones recibidas, vemos que los cuidadores en los que el RN permanece menos tiempo en la UCIN son los que terminan teniendo menos orientaciones sobre cuidados de temperatura, cuidados de higiene y signos de alarma.Conclusión: Los resultados aquí presentados mostraron que la llegada de la familia a casa con el bebé representa una ruptura con el mundo de la hospitalización y genera vivencias del contexto domiciliario.

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).

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.