Variante da Subunidade Beta 3 da Proteína G ( GNB3 ) Está Associada a Alterações Bioquímicas em Pacientes Brasileiros com Hipertensão / G Protein Subunit Beta 3 (GNB3) Variant Is Associated with Biochemical Changes in Brazilian Patients with Hypertension

Resumo Fundamento Genes e suas variantes associadas a fatores ambientais contribuem para o desenvolvimento do fenótipo hipertenso. O gene da subunidade beta 3 da proteína G ( GNB3 ) está envolvido no processo de sinalização intracelular e suas variantes têm sido relacionadas à suscetibilidade à hipertensão arterial. Objetivo Determinar a associação da variante GNB3 (rs5443:C>T) com a hipertensão arterial, parâmetros bioquímicos, idade e obesidade em indivíduos hipertensos e normotensos de Ouro Preto, Minas Gerais. Método A identificação das variantes foi realizada por PCR em tempo real, utilizando o sistema TaqMan®, em amostras de 310 pacientes (155 hipertensos e 155 normotensos). Análises bioquímicas (função renal, perfil lipídico e glicemia) foram realizadas a partir do soro por meio de espectrofotometria UV/Vis e eletrodo íon-seletivo. Foi utilizado um modelo de regressão logística múltipla para identificar fatores associados à hipertensão arterial. A análise das variáveis contínuas com distribuição normal foi realizada usando o teste t de Student não pareado; dados não normais foram analisados usando o teste de Mann-Whitney. Valores de p < 0,05 foram considerados significativos. Resultados A variante rs5443:C>T não esteve associada à hipertensão arterial na população avaliada (p = 0,88). Em relação às medidas bioquímicas, o alelo T esteve associado a níveis elevados de triglicerídeos, glicose e ácido úrico em indivíduos hipertensos (p < 0,05). Conclusão Os presentes resultados mostram a importância do diagnóstico genético para prevenir as causas e consequências de doenças e sugerem que a variante GNB3 rs5443:C>T pode estar associada a alterações no perfil bioquímico em indivíduos hipertensos.

Abstract Background Genes and their variants associated with environmental factors contribute to the development of the hypertensive phenotype. The G protein beta 3 subunit gene (GNB3) is involved in the intracellular signaling process, and its variants have been related to susceptibility to arterial hypertension. Objective To determine the association of the GNB3 variant (rs5443:C>T) with arterial hypertension, biochemical parameters, age, and obesity in hypertensive and normotensive individuals from Ouro Preto, Minas Gerais, Brazil. Method The identification of variants was performed by real-time PCR, using the TaqMan® system, in 310 samples (155 hypertensive and 155 normotensive). Biochemical analyses (renal function, lipid profile and glycemia) were performed from the serum using UV/Vis spectrophotometry and ion-selective electrode. A multiple logistic regression model was used to identify factors associated with arterial hypertension. The analysis of continuous variables with normal distribution was performed using the unpaired Student's t test; non-normal data were analyzed using Mann-Whitney. P < 0.05 was considered significant. Results The rs5443:C>T variant was not associated with arterial hypertension in the evaluated population (p = 0.88). Regarding biochemical measures, the T allele was associated with high levels of triglycerides, glucose and uric acid in hypertensive individuals (p < 0.05). Conclusion These results show the importance of genetic diagnosis to prevent the causes and consequences of diseases and imply that the GNB3 rs5443:C>T variant may be associated with changes in the biochemical profile in hypertensive individuals.

Effects and implications of the COVID-19 pandemic on medicine use by employees of a Brazilian public university: a cross-sectional study

Abstract BACKGROUND: During the COVID-19 pandemic, universities have had to adopt remote education, a strategy that caused sudden changes of routine for everyone involved in academia. OBJECTIVE: To assess the profile of medicine use by the employees of a Brazilian public university during the COVID-19 pandemic. DESIGN AND SETTING: Cross-sectional study at a Brazilian public university. METHODS: Employees were invited to answer an online self-administered questionnaire, containing questions on sociodemographic features, medicine use, mental health and lifestyle habits during the COVID-19 pandemic. The outcome variable was the use of medicines stratified according to occupation. Descriptive, bivariate and multivariate (Poisson regression) statistical analyses were performed. RESULTS: A total of 372 employees participated in the study and use of medicine was reported by 53.2%. Among professors, suicide attempts (prevalence ratio [PR], 1.81; 95% confidence interval [CI], 1.20-2.74), physical activity (PR, 1.53; 95% CI, 1.11-2.11) and poor self-rated health (PR, 1.29; 95% CI, 1.01-1.66); and among technicians, decreased workload during the COVID-19 pandemic (PR, 1.41; 95% CI, 1.00-1.99), excess body weight (PR, 1.39; 95% CI, 1.02-1.88) and poor self-rated health (PR, 1.48; 95% CI, 1.14-1.92) were positively associated with use of medicines. In addition, among technicians, engaging in physical activity (PR, 0.60; 95% CI, 0.46-0.78) was a protective factor against medicine use. CONCLUSION: The profile of medicine use among these employees was similar to that of the Brazilian population. However, some associated factors may have been influenced by the COVID-19 pandemic, thus highlighting the need to examine this topic in a longitudinal study.

Bacterial vaginosis: prevalence, risk profile and association with sexually transmitted infections / Vaginose bacteriana: prevalência, perfil de risco e associação com infecções sexualmente transmissíveis / Vaginosis bacteriana: prevalencia, perfil de riesgo y asociación con infecciones de transmisión sexual

Background and Objectives: Bacterial vaginosis (BV) is the most common cause of vaginal discharge in the world. The study aimed to estimate the prevalence and to identify risk factors associated with bacterial vaginosis. Methods: A cross-sectional study was conducted in Ouro Preto, Brazil, between February and December 2017. Three hundred and forty-one women aged 18 years or older, users of the Brazilian Unified Health System, participated in this study. Women who used oral or topical antibiotics in the four weeks prior to the sample collection and women who had undergone a total hysterectomy were excluded from the study. After signing the Informed Consent Form and filling out a questionnaire containing sociodemographic, behavioral and sexual data, the participants were directed to the collection room, where the nurse collected the samples for the preventive examination of the cervix and also two vaginal swabs. Vaginal swabs and cervical samples were analyzed for cytological abnormalities and BV using Gram staining and cytology. Pathogens causing sexually transmitted infections (STIs) were identified by Polymerase Chain Reaction (PCR). For the analysis of the data, statistical package STATA version 10.0 was used. This study was approved by the Research Ethics Committee of the Federal University of Ouro Preto (UFOP). Results: During the study, 341 women were evaluated. The prevalence of BV using Gram staining (32.5% [CI95% 27.7-37.7%]) and cytology (27.7% [CI95% 23.032.8%]) was similar, however, the sensitivity of cytology was lower (77.8%). Risk factors associated with BV were smoking (IRR 1.5 [CI95%: 1.1  2.1]), use of an intrauterine device (IRR 2.8 [CI95%: 1.2 - 6.5]), and past medical history of BV (IRR 1.5 [CI95%: 1.1 - 2.1]). Correlation between the presence of BV and Trichomonas vaginalis (TV) infection (r=0.24) was observed. Conclusion: The prevalence of BV was affected by life habits and was prevalent in women with TV. Thus, behavioral and social prevention approaches to women with diverse risk profiles may help mitigate TV/BV prevalence and recurrence of BV.(AU)

Contexte et objectifs: La vaginose bactérienne (VB) est la cause la plus fréquente de pertes vaginales dans le monde. Le but de cette étude était d'évaluer la prévalence et les facteurs associés à la vaginose bactérienne. Méthodes: Il s'agit d'une approche descriptive, transversale et quantitative réalisée à Ouro Preto, Minas Gerais, Brésil, entre février et décembre 2017. 341 femmes ont participé à cette étude, âgées de 18 ans ou plus, utilisatrices du Système de santé unifié. Les femmes ayant utilisé des antibiotiques oraux ou topiques dans les quatre semaines précédant le prélèvement et les femmes ayant subi une hystérectomie totale ont été exclues de l'étude. Après avoir signé le formulaire de consentement éclairé et rempli un questionnaire contenant des données sociodémographiques, comportementales et sexuelles, les participants ont été dirigés vers la salle de collecte, où l'infirmière a prélevé les échantillons pour l'examen préventif du col de l'utérus. et aussi deux écouvillons vaginaux. Les échantillons de frottis vaginaux et cervicaux ont été analysés pour les anomalies cytologiques et VB en utilisant la coloration de Gram et la cytologie. Les agents pathogènes causant des infections sexuellement transmissibles (IST) ont été identifiés par réaction en chaîne par polymérase. Pour l'analyse des données, le progiciel statistique STATA version 10.0 a été utilisé. Cette étude a été approuvée par le Comité d'éthique de la recherche de l'Université fédérale d'Ouro Preto (UFOP). Résultats: Au cours de l'étude, 341 femmes ont été évaluées. La prévalence de la VB avec coloration de Gram (32,5% [IC 95% 27,7 - 37,7%]) et de la cytologie (27,7% [IC 95% 23,0 - 32,8%]) était similaire, cependant la sensibilité cytologique était plus faible (77,8%). Les facteurs de risque associés à la VB étaient le tabagisme (IRR 1,5 [IC 95%: 1,1 - 2,1]), l'utilisation d'un dispositif intra-utérin (IRR 2,8 [IC 95%: 1,2 - 6,5] ) et antécédents médicaux de VB (IRR 1,5 [IC 95%: 1,1 - 2,1]). Il y avait une corrélation entre la présence d'une infection à VB et Trichomonas vaginalis (TV) (r = 0,24). Conclusion: La prévalence de la VB était affectée par le mode de vie et l'infection TV. Ainsi, les approches de prévention comportementale et sociale pour les femmes présentant des profils de risque différents peuvent aider à atténuer la prévalence de la TV / VB et la récurrence de la VB.(AU)

Justificativa e Objetivos: A vaginose bacteriana (VB) é a causa mais comum de corrimento vaginal no mundo. O objetivo desse estudo foi avaliar a prevalência e os fatores associados à vaginose bacteriana. Métodos: Trata-se de um descritivo, de forma transversal e abordagem quantitativa realizado em Ouro Preto, Minas Gerais, Brasil, entre fevereiro a dezembro de 2017. Participaram desse estudo 341 mulheres com idade igual ou superior a 18 anos, usuárias do Sistema Único de Saúde (SUS). Mulheres que usaram antibióticos orais ou tópicos nas quatro semanas anteriores à coleta e mulheres que haviam sido submetidas a uma histerectomia total foram excluídas do estudo. Após a assinatura do Termo de Consentimento Livre e Esclarecido e preenchimento de questionário contendo dados sócio-demográfico, comportamental e sexual, as participantes foram encaminhadas para a sala de coleta, onde a enfermeira realizou a coleta das amostras para o exame preventivo do colo do útero e também de dois swabs vaginais. As amostras de esfregaço vaginal e cervical foram analisadas quanto às anormalidades citológicas e VB usando coloração de Gram e citologia. Patógenos causadores de infecções sexualmente transmissíveis (ISTs) foram identificados por Reação em Cadeia da Polimerase (PCR). Para a análise dos dados foi utilizado o pacote estatístico STATA versão 10.0. O presente estudo foi aprovado pelo Comitê de Ética em Pesquisa da Universidade Federal de Ouro Preto (UFOP). Resultados: Durante o estudo, 341 mulheres foram avaliadas. A prevalência de VB com coloração de Gram (32,5% [IC95% 27,7 - 37,7%]) e citologia (27,7% [IC95% 23,0 - 32,8%]) foi semelhante, porém a sensibilidade da citologia foi menor (77,8%). Os fatores de risco associados ao VB foram tabagismo (IRR 1,5 [IC95%: 1,1 - 2,1]), uso de dispositivo intrauterino (IRR 2,8 [IC 95%: 1,2 - 6,5]) e história médica pregressa de VB (IRR 1,5 [IC95%: 1,1 - 2.1]). Observou-se correlação entre a presença de infecção por VB e Trichomonas vaginalis (TV) (r = 0,24). Conclusão: A prevalência de VB foi afetada por hábitos de vida e infecção por TV. Assim, abordagens de prevenção comportamental e social para mulheres com diversos perfis de risco podem ajudar a mitigar a prevalência de TV / VB e recorrência de VB.(AU)

Antibacterial substances from leaves of Protium spruceanum (Burseraceae): in vitro and in silico evaluation

Due to the increase of bacterial resistance, the search for new antibiotics is necessary and the medicinal plants represent its most important source. The aim of this study was to evaluate the antibacterial property of extract and fractions from Protium spruceanum leaves, against pathogenic bacteria. By means of diffusion and microdilution assays, the crude extract was active against the nine bacteria tested being the hydromethanolic fraction the most active. During phytochemical procedures, procyanidin (1) and catechin (2) were identified as the main antibacterial constituents of this fraction. In silico results obtained using PASSonline tool indicated 1 and 2 as having good potential to interact with different targets of currently used antibiotics. These results no indicated potential to none DNA effect and indicated the cell wall as mainly target. Electrophoresis result supported that had no DNA damage. Cell wall damage was confirmed by propidium iodide test that showed increased membrane permeability and by cell surface deformations observed in scanning electronic microscopy. The in vitro assays together with the in silico prediction results establish the potential of P. spruceanum as source of antibacterial compounds that acts on important bacterial targets. These results contribute to the development of natural substances against pathogenic bacteria and to discovery of new antibiotics.

Polymeric micelles containing resveratrol: development, characterization, cytotoxicity on tumor cells and antimicrobial activity

Antimicrobial and antitumor activities of resveratrol, a compound found mainly in grapes, have already been demonstrated. However, its low bioavailability is a limiting factor for therapeutic application. Polymeric micelles can be an approach to solve this problem since they can encapsulate hydrophobic substances. We developed and characterized micellar formulations containing resveratrol and evaluated their cytotoxic and antimicrobial effects. The formulations were prepared by the cold dispersion method with different concentrations of F127 (5 or 10% w/w) and resveratrol (500 or 5000 µM). The formulations were characterized according to size, polydispersity index, pH, encapsulation rate and in vitro release. Cytotoxic effect was evaluated on a bladder cancer cell line and antimicrobial effect was evaluated on E. coli, S. aureus and C. albicans. One of the formulations (10% w/w of F127 and 5000 µM of resveratrol) was a monodispersed solution with high encapsulation rate, thus it was chosen for the cytotoxicity and antimicrobial assays. MS- 10+RES-3 was able to preserve the antimicrobial and cytotoxic activity of resveratrol. This is the first study that evaluated antimicrobial potential and cytotoxicity of micelles containing resveratrol on bladder cancer cells and the results showed that micellar nanostructures could ensure the maintenance of the biological activity of resveratrol.

Increase of binucleated cells in the oral mucosa: a study on the use of psychotropics by students of a Brazilian institution

SUMMARY This study aimed to assess the prevalence of medical and nonmedical use of psychiatric medication among undergraduate students of health sciences from a public university in Brasil. Another objective was to determine the frequency of nuclear morphological abnormalities in the buccal mucosa of students using psychiatric drugs. A cross-sectional study based on a Web survey was carried out with 375 health sciences undergraduate students from schools of Pharmacy, Physical Education, Nutrition, and Medicine. Additionally, spontaneous genetic damages in exfoliated cells of the buccal mucosa of 41 individuals by counting micronucleus (MN) and binucleated (BN) cells frequencies were evaluated. The results showed 76 (20.3%) of students reported the use of psychotropic drugs after enrolling in university. The majority of these students were from Pharmacy and Medicine programs, females, aged between 18-25 years old, nonsmokers, alcohol addicts, and with a family history of mental illness. In addition, Medical students, individuals with high-income, who live alone and are in the last period of the program are more likely to use psychotropic drugs. Moreover, exposure to psychiatric medication was able to increase the number of binucleated cells. These results provide evidence that the use of psychoactive drugs is increased in the academic context and may be related to the failure of the cell cycle.

RESUMO O objetivo deste estudo foi avaliar a prevalência de uso médico e não médico de medicação psiquiátrica entre estudantes de graduação em Ciências da Saúde de uma universidade pública do Brasil. Outro objetivo foi determinar a frequência de anormalidades nucleares morfológicas na mucosa bucal desses estudantes. Realizou-se um estudo transversal baseado na Web com 375 estudantes de graduação em Ciências da Saúde das escolas de Farmácia, Educação Física, Nutrição e Medicina. Adicionalmente, danos genéticos espontâneos em células esfoliadas da mucosa bucal de 41 indivíduos foram analisados para estudo da frequência de micronúcleos e células binucleadas. Os resultados mostraram que 76 (20,8%) dos estudantes relataram que usaram drogas psicotrópicas após se matricularem na universidade. O uso desses medicamentos foi maior entre estudantes de cursos de Farmácia e Medicina, sexo feminino, idade entre 18 e 25 anos, não tabagistas, alcoolistas e histórico familiar de doença mental. Além disso, estudantes de Medicina, indivíduos com alta renda, que moram sozinhos e estão no último período do curso são mais propensos a usar drogas psicotrópicas. Ademais, a exposição à medicação psiquiátrica foi capaz de aumentar o número de células binucleadas. Esses resultados fornecem evidências de que o uso de drogas psicotrópicas aumenta no contexto acadêmico e pode estar relacionado à falha do ciclo celular.

Anti-serum validation for use in immunohistochemistry for Trypanosoma cruzi detection

Abstract INTRODUCTION: The detection of Trypanosoma cruzi in tissue samples is important in many situations, such as testing of the reactivation of the infection. The detection of T. cruzi nests in endomyocardial biopsies (EMB) may be useful to evaluate graft rejection. Given their scarcity, such nests are not routinely identified. To increase the diagnosis sensitivity, immunohistochemistry (IHC) may serve as a promising strategy. Here, we validate an antiserum for the detection of T. cruzi infection by IHC. METHODS: We used 1) positive controls (PCs) - 13 EMB, 12 skin biopsies, and 1 heart with T. cruzi nests as sections stained with hematoxylin and eosin (HE); 2) negative controls - a) 10 explant hearts and 10 EMB with no amastigote nests or clinical/laboratory signs of chagasic infection; and b) eight samples with leishmaniasis, toxoplasmosis, or histoplasmosis; and 3) Cases - 31 EMB of chagasic patients with no parasite nests in HE sections but detected positive for T. cruzi DNA by polymerase chain reaction. As a primary antibody, a hyperimmune serum from T. cruzi-infected rabbits was used. RESULTS: IHC results were positive for 21 of 26 PCs (80.8%) and one case of cutaneous leishmaniasis. In 4 of 31 cases, IHC revealed nests (12.9%), which were undetected by conventional histological examination. CONCLUSIONS: This study shows that IHC with the tested antiserum increases the sensitivity of the diagnosis and may be recommended for routine use in EMB analyses of cardiac transplant patients with Chagas disease.

Microdissecção e captura a laser na investigação do gene TP53 em tecidos incluídos em parafina / Laser-capture microdissection for TP53 gene analysis in paraffin-embedded tissues

INTRODUÇÃO: Microdissecção e captura a laser (MCL) é uma técnica de desenvolvimento recente que permite a coleta de células individuais ou pequeno conjunto de células para análise molecular. Atualmente, no Brasil, há raros microscópios para MCL, de modo que a divulgação dos procedimentos inerentes a essa técnica é oportuna para destacar seu amplo potencial para diagnóstico e investigação. OBJETIVO: Este trabalho descreve a padronização dos procedimentos de MCL e de extração de DNA de material fixado em formalina e incluído em parafina. MATERIAL E MÉTODOS: Foram estudados o éxon 8 do gene TP53 e o gene da ciclofilina em amostras de tecido normal e de neoplasias de fígado e rim provenientes de modelo de carcinogênese química induzida em rato. A extração do DNA foi comprovada por reação em cadeia da polimerase (nested-PCR). RESULTADOS: Foram padronizados os procedimentos de preparo dos cortes histológicos, de microdissecção e captura a laser e de obtenção de seqüências gênicas pela reação de nested-PCR para tecidos incluídos em parafina. Obtivemos amplificação de 48,3 por cento das amostras para o éxon 8 do gene TP53 e 51,7 por cento para o gene da ciclofilina. Considerando pelo menos um dos dois segmentos gênicos, foram amplificadas 79,3 por cento das amostras. DISCUSSÃO E CONCLUSÃO: A extração de DNA de tecidos fixados em formalina e incluídos em parafina e a técnica de nested-PCR foram adequadamente padronizadas para produtos gênicos de interesse, obtidos de material coletado por MCL. Esses procedimentos podem ser úteis para a obtenção de seqüências de DNA de arquivos para análise molecular.

BACKGORUND: Laser-capture micro-dissection (LCM) is a recently developed procedure that provides single cells or specific cell groups for molecular analysis. Currently, there are few LCM systems in Brazil, in such a way that it is necessary to disseminate the technical procedures inherent to the methodology, and also to characterize its enormous potential for diagnosis and research. OBJECTIVE: This study describes the standardization of LCM and DNA extraction from formalin fixed and paraffin-embedded tissues. MATERIAL AND METHOD: The gene TP53 exon 8 and the cyclophilin gene were studied in normal and neoplastic liver and kidney samples from a chemical carcinogenesis model in rat. DNA extraction was confirmed by nested-PCR. RESULTS: Histological sections preparation for LCM and the nested-PCR procedures were standardized; 48.3 percent amplifications of the gene TP53 exon 8 and 51.7 percent of the cyclophilin gene samples were obtained. When at least one of the gene segments was considered, 79.3 percent samples presented amplification. DISCUSSION AND CONCLUSION: Procedures for DNA extraction from formalin fixed and paraffin-embedded tissues collected by LCM were standardized. They can be useful for DNA collection for molecular studies.