A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17.

This manuscript describes a unique protocol for the rapid transformation of Medicago truncatula A17 cell suspension cultures mediated by Agrobacterium tumefaciens. Medicago cells were collected on day 7 of the growth curve, which corresponded to the beginning of the exponential phase. They were then co-cultured with Agrobacterium for 3 days before being spread onto a petri dish with appropriate antibiotic selection. The Receptor Binding Domain of the Spike protein of SARS-CoV-2 was used as a model to develop this protocol. The presence of the transgene was assessed using PCR, and the integrity of the product was evaluated by SDS-PAGE and Western-blotting.

Production of the SARS-CoV-2 Spike protein and its Receptor Binding Domain in plant cell suspension cultures.

The COVID-19 pandemic, caused by the worldwide spread of SARS-CoV-2, has prompted the scientific community to rapidly develop efficient and specific diagnostics and therapeutics. A number of avenues have been explored, including the manufacture of COVID-related proteins to be used as reagents for diagnostics or treatment. The production of RBD and Spike proteins was previously achieved in eukaryotic cells, mainly mammalian cell cultures, while the production in microbial systems has been unsuccessful until now. Here we report the effective production of SARS-CoV-2 proteins in two plant model systems. We established transgenic tobacco BY-2 and Medicago truncatula A17 cell suspension cultures stably producing the full-length Spike and RBD recombinant proteins. For both proteins, various glycoforms were obtained, with higher yields in Medicago cultures than BY-2. This work highlights that RBD and Spike can be secreted into the culture medium, which will impact subsequent purification and downstream processing costs. Analysis of the culture media indicated the presence of the high molecular weight Spike protein of SARS-CoV-2. Although the production yields still need improvement to compete with mammalian systems, this is the first report showing that plant cell suspension cultures are able to produce the high molecular weight Spike protein. This finding strengthens the potential of plant cell cultures as production platforms for large complex proteins.

Prevalência de tromboembolismo pulmonar diagnosticado por angiotomografia computadorizada em pacientes de um município de médio porte de Minas Gerais / Prevalence of pulmonary thromboembolism diagnosed by computadorized angiotomography in patients in a inner city in state of Minas Gerais

Introdução. O tromboembolismo pulmonar (TEP) é uma doença que apresenta uma clínica inespecífica, o que induz a necessidade de solicitar exames complementares, como a angiotomografia computadorizada (angioTC) de tórax. Com o objetivo de evitar a sua solicitação desnecessária, foram criados sistemas de probabilidade pré-teste, como o escore de Wells, que direcionam a propedêutica. Objetivo. Avaliar a prevalência de tromboembolismo pulmonar (TEP) em dois hospitais de uma cidade de médio porte e verificar o padrão de solicitação da angiotomografia computadorizada de tórax (angioTC). Métodos. Estudo transversal a partir de dados de prontuários de pacientes submetidos à angioTC de tórax no Hospital Regional de Barbacena e no Hospital Ibiapaba no período de janeiro de 2014 até junho de 2018. Resultados. A prevalência de TEP foi de 29,70% (N=259) no período de 54 meses e a mediana de idade, 66 anos (intervalo interquartil: 24). Nos pacientes com o diagnóstico de TEP, o sintoma mais frequente foi dor torácica (38,81%), seguido por hemoptise (33,33%) e tosse (32,99%). Nos prontuários analisados foi observado que a utilização do escore de Wells foi de 2,70%. A probabilidade de TEP, pelo escore de Wells, realizada pelos pesquisadores com base nos dados dos prontuários, em pacientes diagnosticados com TEP pela angioTC foi alta em 45%, moderada em 34,42% e baixa em 9,23%. Conclusão. A prevalência foi de 29,70% no período de 54 meses de diagnóstico de TEP em pacientes que realizaram a angioTC de tórax nos hospitais mencionados. O escore de Wells é um importante instrumento propedêutico. (AU)

Background. Pulmonary embolism (PE) is a disease presented with nonspecific symptoms that requires additional tests such as pulmonary angiography by computadorized tomography. To avoid unnecessary tests, pre-test probability tools were created, such as Wells Score, which guides the propaedeutics. Objective. To evaluate the prevalence of pulmonary embolism (PE) in two hospitals in a medium-sized city and to verify computed tomography pulmonary angiography imaging exam orders. Methods. Cross-sectional study based on data from the hospital records evolving patients underwent computed tomography pulmonary angiography at Hospital Regional de Barbacena and Hospital Ibiapaba through January 2014 and June 2018. Results. The prevalence of PE was 29.70% (259 patients) in the 54-month period and the median age, 66 years (interquartile range, 24). In patients with the diagnosis of PE, the most frequent symptom was chest pain (38.81%) followed by hemoptysis (33.33%), and cough (32.99%). In the analyzed hospital records it was observed that the use of the Wells score was 2.70%. The probability of PE according to the Wells score, performed by the researchers based on data from the medical records, in patients diagnosed with PE by computed tomography pulmonary angiography was high in 45%, moderate in 34.42%, and low in 9.23%. Conclusion. The prevalence of PE was 29.70% (N = 259) in the 54-month period from PE diagnosis in patients who underwent computed tomography pulmonary angiography in the mentioned hospitals. The Wells score is an important diagnostic tool(8)

Avaliação das atividades instrumentais de vida diária dos idosos em inquérito domiciliar / Evaluation of the instrumental activities of daily life in the household survey

O presente estudo teve como objetivo caracterizar os idosos quanto ao comprometimento da capacidade funcional para realização das AIVD viventes na cidade de Picos - Piauí. Trata-se de um estudo de corte transversal, descritivo e abordagem quantitativa com amostragem probabilística complexa, por conglomerados, em 2 estágios: setores censitários e domicílios. O estudo foi composto por 67 idosos de ambos os sexos com idades a partir de 60 anos; a amostragem foi aleatória por conveniência, com idosos que estivessem contemplados dentro dos setores censitários do lote 02 e 05. As variáveis foram agrupadas em dados pessoais e socioeconômicos (idade, sexo, estado civil, escolaridade, renda mensal, domicilio, ocupação atual e pessoas com quem reside), variáveis clínicas e hábitos de vida (doenças diagnosticadas, utilização de medicamentos, prática de atividades físicas, tabagismo e alcoolismo) e variáveis relacionadas às atividades instrumentais de vida diária de Lawton (utilizar medicamentos, fazer compras, preparar refeições, utilizar telefone sozinho (a), gerenciar tarefas domésticas e lidar com transportes e finanças). Nesse estudo foi possível observar a influência das variáveis: sexo, idade, estado civil, escolaridade, ocupação atual, renda e atividade física, que se relacionam diretamente com a necessidade de auxílio para a realização das AIVD. A população estudada manteve sua independência para as AIVD, principalmente em relação ao preparo de suas refeições (82,1%) e ingestão de medicamentos (80,6%). Em relação ao nível de dependência dos participantes da pesquisa, existe uma porcentagem significativa de 43,28% de independência de idosos com idades entre 60 e 70 anos, e apenas 2,99% desses idosos são dependentes nas atividades instrumentais de vida diária. Os idosos que possuem idade maior ou igual a 80 anos apresentam 2,99% de independência e 4,47% de dependência. Dessa forma, a obtenção de conhecimento acerca dos fatores associados à manutenção da capacidade funcional do idoso é imprescindível para subsidiar políticas públicas que favoreçam a saúde integral dessa população, detectando precocemente condições que possam gerar incapacidades

The present study aimed to evaluate the ability to perform instrumental activities of daily living (IADL) in the elderly. This research is part of the project called "Population-based health survey in municipalities of Piauí", developed through a partnership between the Federal University of Piauí (UFPI) with the University of São Paulo (USP). This is a cross-sectional, descriptive and quantitative-approach study. The data collected were entered and analyzed through the STATISTICAL PROGRAM IBM Statistical Package for the Social Sciences (SPSS), version 20.0. In this study, it was possible to observe the influence of the variables gender, age, marital status, schooling, current occupation, income and physical activity that directly relate to the need for assistance in performing (IADL). Thus, obtaining knowledge about the factors associated with maintaining the functional capacity of the elderly is essential to support public policies that favor the integral health of this population and the care offered to it, stimulating a aging and detecting early conditions that can generate disabilities.

Escuta terapêutica: uma tecnologia do cuidado em saúde mental / Therapeutic listening: a technology of mental health care

Objetivo: caracterizar a escuta terapêutica analisando os benefícios na atenção a pessoas com transtornos mentais em diferentes níveis assistenciais. Método: trata-se de um estudo bibliográfico, descritivo, tipo revisão integrativa da literatura, entre 2010 a 2020, na LILACS, MEDLINE, BVS e BDENF. Analisaram-se os artigos pela leitura reflexiva e criteriosa acerca das principais informações e elementos que compõem a temática nos estudos. Resultados: resultaram-se 12 artigos, publicados em periódicos nacionais. Aponta-se a escuta terapêutica como uma importante ferramenta para a análise mais favorável ao entendimento do real sofrimento psíquico do paciente, valorizando as relações profissional-paciente-família e, além disso, necessitando do desenvolvimento de habilidades técnicas e humanísticas, favorecendo uma autorreflexão do paciente e proporcionado uma rede de cuidados instrumentalizada e sistematizada. Conclusão: conclui-se que a escuta é uma eficiente tecnologia terapêutica que pode e deve ser implementada em diversos cenários, mas que carece de maiores pesquisas acerca do impacto nos diferentes níveis, requerendo o desenvolvimento de habilidades técnicas para sua melhor operação.(AU)

Objective: to characterize therapeutic listening by analyzing the benefits in care for people with mental disorders at different levels of care. Method: this is a bibliographic, descriptive, integrative literature review, between 2010 and 2020, at LILACS, MEDLINE, BVS and BDENF. The articles were analyzed by reflective and careful reading about the main information and elements that make up the theme in the studies. Results: 12 articles resulted, published in national journals. Therapeutic listening is pointed out as an important tool for the analysis more favorable to the understanding of the patient's real psychological suffering, valuing the professional-patient-family relationships and, in addition, requiring the development of technical and humanistic skills, favoring a self-reflection of the patient and provided an instrumented and systematized care network. Conclusion: it is concluded that listening is an efficient therapeutic technology that can and should be implemented in different scenarios, but that lacks further research on the impact at different levels, requiring the development of technical skills for its best operation.(AU)

Objetivo: caracterizar la escucha terapéutica mediante el análisis de los beneficios en la atención de personas con trastornos mentales en diferentes niveles de atención. Método: se trata de una revisión bibliográfica, descriptiva, integradora de la literatura, entre 2010 y 2020, en LILACS, MEDLINE, BVS y BDENF. Los artículos fueron analizados mediante una lectura reflexiva y cuidadosa sobre la información principal y los elementos que componen el tema en los estudios. Resultados: resultaron 12 artículos, publicados en revistas nacionales. La escucha terapéutica se señala como una herramienta importante para el análisis más favorable a la comprensión del sufrimiento psicológico real del paciente, valorando las relaciones profesionales-paciente-familiares y, además, requiere el desarrollo de habilidades técnicas y humanísticas, favoreciendo una autorreflexión del paciente y proporcionando una red de atención instrumentada y sistematizada. Conclusión: se concluye que escuchar es una tecnología terapéutica eficiente que puede y debe implementarse en diferentes escenarios, pero que necesita más investigación sobre el impacto a diferentes niveles, lo que requiere el desarrollo de habilidades técnicas para su mejor operación.(AU)

Effect of ultrafiltration on placental-fetal blood flow in pregnancy of woman undergoing chronic hemodialysis.

INTRODUCTION: Patient who was undergoing hemodialysis (HD) thrice weekly usually gain 1 to 4 kg of weight in interdialytic period, mainly due to fluid accumulation by ingestion of water. Ultrafiltration (UF) during HD will be need to remove fluid excess to avoid severe medical complications secondary to fluid overload. However, in pregnant woman UF can increase the episodes of intradialytic hypotension which may lead to placental ischemic injury and predispose to fetal distress. There is little information about safe fluid amount withdrawn by UF during pregnancy. METHODS: We prospectively study by obstetric Doppler ultrasonography the fluxometric parameters: pulsatility index (PI) and resistance index (RI) of fetal middle cerebral, uterine, and umbilical artery obtained at the beginning and the end of HD session, the acute and chronic effect of UF on placenta and fetus blood flow, as well as the fetal outcome in 1 pregnant woman on chronic HD. FINDINGS: We did not observe any acute harmful effect on fetal middle cerebral, placental and umbilical artery blood flow when UF rate of 2.1 ± 0.04 L (6 < 8 mL/h/kg) during HD session, no significant statistical difference was observed when compared PI and RI before and after UF and also when we compared these data with reference value on normal pregnancy to the same gestational age. DISCUSSION: UF rate of 6 < 8 mL/h/kg during HD did not bring any acute harmful effect on fetal middle cerebral, placental, and umbilical blood flow and the UF rate of 1.4 6 0.4 L (< 6 mL/h/kg) / HD session that was done in all others HD during pregnancy was safe, without any chronic fetal deleterious effect. Obstetric Doppler ultrasonography is a simple and noninvasive method to fetal follow-up and can aid to determine safe UF rate in pregnant women during gestation.

Ecology of the non-native snail Sinotaia cf quadrata (Caenogastropoda: Viviparidae). A study in a lowland stream of South America with different water qualities.

Sinotaia quadrata is a snail native from Asia recorded for the first time in South America in 2009 in central Argentina. In 2015, this species was also found in a lowland stream with different water qualities. Our aims were to contribute to the knowledge of its population ecology and to compare the individuals from the two locations anatomically. Snails were searched at 6 sites, where physicochemical and hydraulic parameters were measured. Biological samples were also taken at two sites (S3 and S4) to study the population traits of S. cf quadrata (density, size structure, fecundity and sex ratio) and to assess the water quality through macroinvertebrates' biological indices (richness, diversity and IBPamp). Physicochemical and biological parameters allowed us classifying sites as "moderately polluted" (S3) and "heavily polluted" (S4). At S4, the population showed a lower density, larger individuals, higher fecundity and a scarce representation of young snails. The differences observed in the radula and mantle border of snails from the two geographical regions might be attributed to environmental differences. We conclude that this species is tolerant to a wide range of environmental variables which, along with its high fecundity and morphological plasticity, could allow this species to colonize neighbor streams.

Ecology of the non-native snail Sinotaia cf quadrata (Caenogastropoda: Viviparidae). A study in a lowland stream of South America with different water qualities

ABSTRACT Sinotaia quadrata is a snail native from Asia recorded for the first time in South America in 2009 in central Argentina. In 2015, this species was also found in a lowland stream with different water qualities. Our aims were to contribute to the knowledge of its population ecology and to compare the individuals from the two locations anatomically. Snails were searched at 6 sites, where physicochemical and hydraulic parameters were measured. Biological samples were also taken at two sites (S3 and S4) to study the population traits of S. cf quadrata (density, size structure, fecundity and sex ratio) and to assess the water quality through macroinvertebrates' biological indices (richness, diversity and IBPamp). Physicochemical and biological parameters allowed us classifying sites as "moderately polluted" (S3) and "heavily polluted" (S4). At S4, the population showed a lower density, larger individuals, higher fecundity and a scarce representation of young snails. The differences observed in the radula and mantle border of snails from the two geographical regions might be attributed to environmental differences. We conclude that this species is tolerant to a wide range of environmental variables which, along with its high fecundity and morphological plasticity, could allow this species to colonize neighbor streams.