Quantification of in vitro replication kinetics of Alagoas vesiculovirus isolates by digital droplet RT-PCR.

Vesicular stomatitis caused by Alagoas vesiculovirus (VSAV) has generated disease outbreaks in Brazil, mainly in the northeast region. Phylogenetic studies divide the isolates into three distinct genotypes (A, B, and C). However, there is no description of how this genetic divergence reflects on the phenotype of VSAV isolates such as in vitro replication fitness. Therefore, the objective of this work was to evaluate the ability of three distinct genotypes of Brazilian isolates of VSAV to grow in different cell-culture lines (BHK-21, Vero, and NCI-H1299). Quantification of viral RNA was performed using RT-PCR digital droplet from supernatant of cell culture collected every 4 h for a period of 24 h of viral growth in three different cell lines (BHK-21, Vero, and NCI-H1299). It was observed that the genotype C isolate has the lowest replication efficiency among the three analyzed viruses, without major changes in the copies of viral RNA over the entire time of the study.

Estratégias resilientes de jogadores de futebol profissional atuantes no estado do Ceará / Resilient strategies of professional soccer players acting in the state of Ceará / Estrategias resistentes de jugadores de fútbol profesionales que actúan en el estado de Ceará

Esta pesquisa objetiva investigar a existência de comportamentos resilientes em jogadores de futebol profissional atuantes no estado do Ceará, bem como identificar estratégias de enfrentamento e redes de apoio social dos jogadores. O estudo é fruto de uma pesquisa descritiva exploratória, qualitativa, na qual foram entrevistados dez jogadores de futebol vinculados a clubes cearenses, entre 18 e 38 anos, nascidos no estado ou não. Para contatá-los foi utilizado as redes sociais, o método bola de neve e o auxílio das assessorias esportivas. Devido a pandemia, a coleta de dados ocorreu de forma online e estes foram analisados através de uma análise de conteúdo. Os resultados mostraram que os jogadores entrevistados são munidos de características da personalidade resiliente, tais como: crença em seu próprio potencial, o diálogo com pessoas de confiança, a religiosidade, as motivações individuais e o foco nos objetivos, e possuem uma forte rede de apoio social. Pôde-se concluir que os jogadores detêm um alto nível de resiliência, utilizando-se destas estratégias para superarem as dificuldades e se fortalecerem. Evidenciou-se que na percepção dos jogadores a atuação do psicólogo no esporte é crucial.

This research aims to analyze whether there is resilience in professional soccer players working in the state of Ceará, describe the resilient strategies used and identify whether players have a social support network. The study is the result of an exploratory, qualitative descriptive research, in which ten football players linked to Ceará clubs, between 18 and 38 years old, born in the state or not, were interviewed. To contact them, social networks, the snowball method and the help of sports advisors were used. Due to the pandemic, data collection took place online and these were analyzed through content analysis. The results showed that the interviewed players are equipped with resilient personality characteristics, such as: belief in their own potential, dialogue with trusted people, religiosity, individual motivations and focus on goals, and have a strong support network. Social. It can be concluded that players have a high level of resilience, using these strategies to overcome difficulties and strengthen themselves. It was evidenced that in the perception of the players the role of the psychologist in the sport is crucial.

Esta investigación tiene como objetivo analizar si existe resiliencia en futbolistas profesionales que actúan en el estado de Ceará, describir las estrategias resilientes utilizadas e identificar si los jugadores cuentan con una red de apoyo social. El estudio es resultado de una investigación cualitativa descriptiva, exploratoria, en la que fueron entrevistados diez futbolistas vinculados a clubes de Ceará, entre 18 y 38 años, nacidos en el estado o no. Para contactarlos se utilizaron las redes sociales, el método bola de nieve y la ayuda de asesores deportivos. Debido a la pandemia, la recolección de datos se realizó en línea y estos fueron analizados a través del análisis de contenido. Los resultados mostraron que los jugadores entrevistados están dotados de características de personalidad resilientes, tales como: creencia en su propio potencial, diálogo con personas de confianza, religiosidad, motivaciones individuales y enfoque en objetivos, y cuentan con una fuerte red de apoyo social. Se puede concluir que los jugadores tienen un alto nivel de resiliencia, utilizando estas estrategias para superar las dificultades y fortalecerse. Se evidenció que en la percepción de los jugadores el papel del psicólogo en el deporte es crucial.

Outbreaks of Vesicular Stomatitis in Brazil caused by a distinct lineage of Alagoas vesiculovirus.

This article describes the recurrence of outbreaks of Vesicular Stomatitis in the State of Maranhão, Brazil. The procedures for treating the outbreak of vesicular disease, sample collection, laboratory tests performed, and the results obtained were described. The clinical signs and observed injuries have been described. The sera showed antibodies that cross-react between the Vesiculovirus Indiana, Cocal, and Alagoas. The serological profile shows the presence of high antibody titers for Alagoas vesiculovirus in cattle, swine, and horses. Higher antibody titers indicate the viral serotype present in the outbreak. The genetic sequencing of the isolates confirmed the presence of Alagoas vesiculovirus, which grouped with the virus isolated in 2013 from cattle from the State of Maranhão.

Práticas de terapia ocupacional na rede de saúde mental da criança e do adolescente / Occupational therapy practices in the child and adolescent mental health network

Resumo Introdução A rede de atenção à saúde mental da criança e do adolescente se expandiu, nos últimos anos, tornando-se espaço de ação significativo do terapeuta ocupacional. Objetivo Caracterizar as práticas dos terapeutas ocupacionais na rede de atenção à saúde mental infantojuvenil do município de Belo Horizonte, MG, identificando ações e abordagens. Método Estudo qualitativo, exploratório-descritivo, realizado com os terapeutas ocupacionais da rede de atenção do município. A coleta de dados se deu por meio de entrevistas semiestruturadas e o recurso para seu tratamento foi a análise de conteúdo temática. Resultados Os terapeutas ocupacionais estão presentes nos três dispositivos da rede de assistência: nove nas Equipes Complementares; cinco nos CAPSi e quatro no Programa Arte da Saúde. Nas Equipes Complementares, identificaram-se três perspectivas de abordagem: a desenvolvimentista/habilitadora; a de tipo psicoterápico; e a perspectiva fundada nos princípios da reabilitação psicossocial. Nos CAPSi, destaca-se a similaridade das ações dos membros da equipe, sem especificação de práticas baseadas nos núcleos profissionais, o que repercute, com tensões, sobre a identidade profissional. No Programa Arte da Saúde, os terapeutas ocupacionais, coordenadores deste serviço, exercem a função de gestão. Conclusão Verificou-se que o terapeuta ocupacional tem participado e contribuído na consolidação da rede de assistência do município e de suas propostas. Identificou-se que não há práticas homogêneas e que estas se relacionam às características e objetivos dos dispositivos e às especificidades da inserção do profissional. Por outro lado, identificaram-se convergências de práticas e abordagens, possibilitando o delineamento de algumas ações do profissional no campo.

Abstract Introduction The network of mental healthcare for children and adolescents has expanded in recent years, becoming a significant space for occupational therapists' actions. Objective To characterize the practices of occupational therapists in the children's mental healthcare network in Belo Horizonte, identifying actions and approaches. Method Qualitative, exploratory-descriptive study carried out with occupational therapists in the care network. The data collection took place trough semi-structured interviews and resource for their prossessing was thematic content analysis. Results Occupational therapists are present in the three devices of assistance network: nine in Complementary Teams; five in the CAPSi; four in the Art of Health Program. In the Complementary Teams, we identified three perspectives: the psychotherapeutic approach, the developmental/enabler approach and the perspective one, based on the principles of psychosocial rehabilitation. In CAPSi, the similarity of the team members' actions stands out, without specifying practices based on the professional nuclei, with affects, with tensions, the professional identity. In the Art of Health Program, occupational therapists, coordinators of this service, exercise the management function. Conclusion The occupational therapists participate and contribute to the consolidation of proposals and the municipality's assistance network. There are no homogeneous practices. Such actions depend on the characteristics and scope of the service and the specificities of the professional's insertion. On the other hand, we identified the convergence of practices and approaches, making it possible to outline the professional's actions in the field.

[Combined HIV prevention? Systematic review of interventions with women from low- and middle-income countries]. / Prevenção combinada do HIV? Revisão sistemática de intervenções com mulheres de países de média e baixa renda.

The scope of this systematic review was to characterize interventions for HIV prevention that included adult women and was conducted in low- and middle-income countries after 1996, identifying how they covered individual, social and programmatic vulnerabilities. Databases were accessed from 1997 to July 2016 and the studies selected included adult women, with combined strategies or otherwise, only excluding biomedical or mass media interventions. Thus, 72 interventions developed in 32 countries were selected, most of them in African countries and/or in the upper-middle income bracket, with 26 focusing solely on women. Among the 64 interventions that evaluated results in behavioral, psychosocial and biomedical knowledge on HIV/STI and use of health services, 62 reported positive results, but also of no-effect (n = 52). Few of them reported a reduction in HIV/STI incidence (n = 9), in partner numbers (n = 12) and in stigma and violence reports (n = 7). The content analysis of the interventions revealed that the majority (n = 51) considered only individual vulnerabilities. The need to strengthen preventive approaches targeting structural determinants of the epidemic, encompassing the social contexts of women in a relational and intersectional perspective of vulnerabilities was discussed.

RT-qPCR for the diagnosis of the vesiculovirus Cocal virus.

Cocal virus (COCV) is one of the causative agents of vesicular stomatitis, presenting clinical signs indistinguishable from those caused by foot-and-mouth disease virus (FMDV). Therefore, the differentiation of these two viruses via laboratory diagnosis is essential. The objective of this study was to develop and validate a real-time quantitative PCR (RT-qPCR) protocol for the diagnosis of COCV directly from epithelial samples. The method developed had 97% accuracy at 3950 pfu and a repeatability error of 1.29%. RT-qPCR was able to distinguish COCV from other viruses that cause vesicular diseases, an important factor because seroneutralization may produce cross-reactivity between COCV and vesicular stomatitis Alagoas virus (VSAV). No epithelial sample originating from vesicular disease outbreaks between 2014 and 2018 in Brazil was positive for COCV.

Prevenção combinada do HIV? Revisão sistemática de intervenções com mulheres de países de média e baixa renda / Combined HIV prevention? Systematic review of interventions with women from low- and middle-income countries

Resumo A revisão sistemática objetivou caracterizar as intervenções para prevenção do HIV que incluíam mulheres adultas e desenvolvidas em países de baixa e média renda após 1996, identificando como abarcavam as vulnerabilidades individuais, sociais e programáticas. Foram acessadas bases de dados de 1997 a julho de 2016 e selecionados estudos que incluíam mulheres adultas, com estratégias combinadas ou não, excluindo-se intervenções apenas biomédicas ou de mídia em massa. Foram selecionadas 72 intervenções desenvolvidas em 32 países, sendo a maioria africanos e/ou de renda média-alta e 26 só com mulheres. Entre 64 intervenções que avaliaram aspectos comportamentais, psicossociais, biomédicos, de conhecimentos sobre HIV/IST e uso de serviços de saúde, 62 tiveram resultados positivos, mas também de não efeito (n = 52). Poucas relataram diminuição na incidência de HIV/IST (n = 9), no número de parceiros (n = 12), dos relatos de estigma e de violências (n = 7). A análise de conteúdo das intervenções mostrou que a maioria (n = 51) contemplava apenas vulnerabilidades individuais. Discute-se a necessidade de fortalecer abordagens preventivas que atuem nos determinantes estruturais da epidemia, abarcando os contextos sociais das mulheres numa perspectiva relacional e interseccional das vulnerabilidades.

Abstract The scope of this systematic review was to characterize interventions for HIV prevention that included adult women and was conducted in low- and middle-income countries after 1996, identifying how they covered individual, social and programmatic vulnerabilities. Databases were accessed from 1997 to July 2016 and the studies selected included adult women, with combined strategies or otherwise, only excluding biomedical or mass media interventions. Thus, 72 interventions developed in 32 countries were selected, most of them in African countries and/or in the upper-middle income bracket, with 26 focusing solely on women. Among the 64 interventions that evaluated results in behavioral, psychosocial and biomedical knowledge on HIV/STI and use of health services, 62 reported positive results, but also of no-effect (n = 52). Few of them reported a reduction in HIV/STI incidence (n = 9), in partner numbers (n = 12) and in stigma and violence reports (n = 7). The content analysis of the interventions revealed that the majority (n = 51) considered only individual vulnerabilities. The need to strengthen preventive approaches targeting structural determinants of the epidemic, encompassing the social contexts of women in a relational and intersectional perspective of vulnerabilities was discussed.

Effect of smoking on the DNA methylation pattern of the SOCS1 promoter in epithelial cells from the saliva of patients with chronic periodontitis.

BACKGROUND: The aim of the present study was to evaluate the methylation pattern in the suppressor of cytokine signaling 1 (SOCS1) gene in smokers and non-smokers with chronic periodontitis (CP). METHODS: Methylation-specific polymerase chain reaction (PCR) was performed to determine the methylation status of the SOCS1 promoter in 45 saliva samples from smokers and non-smokers with CP. RESULTS: Cells from the saliva of CP patients who smoked were 7.08 times more likely to have a methylated SOCS1 promoter than cells from the saliva of non-smoking patients. CONCLUSIONS: SOCS1 gene promoter methylation, with its potential effects on the expression of this gene, seems to be a consequence of exposure to tobacco and not to periodontal disease. Further studies are needed to elucidate the relationship between the epigenetic control of immune response gene expression, exposure to environmental factors, and the development, progression, and prognosis of CP.

Resilience in the Training of Medical Students in a University With a Hybrid Teaching-Learning System / Resiliencia en la Formación de Estudiantes de Medicina en una Universidad con un Sistema Híbrido de Ensenãnza y Aprendizaje

ABSTRACT Background The exposure of students to stressful events and the association of these events with students' mental health is an important matter in Medical Education. To address this arduous training and solve emerging problems, some students develop methods to help them and, among these, resilience. A hybrid learning system, merging active and traditional learning, can be a supplementary source of stress generation , since it demands the acquisition of knowledge by the students, for summative assessments of traditional teaching as well as for the autonomous search for knowledge, skills, and attitudes required in the problematization. Purpose To determine the degree of resilience throughout the medical course under the hybrid teaching-learning system, identifying underlying mechanisms. Methods This was a cross-sectional study developed from August 2017 to August 2018, at Christus University Center, Brazil, a medical school that uses problem-based learning curricula associated with traditional teaching methodology. Wagnild and Young Resilience Scale was applied to medical students from all semesters. Socioeconomic, emotional and self-reported performance variables were also collected. The association between variables was assessed with minimally adjusted logistic regression models. Results 173 medical students participated in this study, with a mean age of 22.4 years, of which 65.3% were females. 88.1% of the medical students showed high or very high resilience. Receiving support from family and friends was associated with better resilience (p values lower than 0.001), as students who were "very satisfied" or "satisfied" with family support had a greater tendency to develop better degrees of resilience, with results of "very high resilience trends" (82.50%) and "high resilience trends" (71.10%) surpassing the prevalence identified in dissatisfied students. Also, having a religious belief was also associated with higher resilience degrees (p value = 0.02). Conclusions Factors identified in this study, mainly the importance of the support network from family and friends can be stimulated in order to improve students' resilience. There was no direct association between the academic performance self-assessment and the students' resilience and the resilience of medical students tends to remain constant throughout the course.

RESUMEN Introducción La exposición de estudiantes a eventos que los estresan y la asociación de esos eventos con la salud mental de los alumnos, es una cuestión importante en la Educación Médica. Para andar por el arduo camino del pregrado y resolver problemas emergentes, algunos alumnos desarrollan métodos que los ayuden, entre ellos, la resiliencia. Un sistema de aprendizaje híbrido, con fusión de aprendizaje activo y tradicional, puede ser un sistema complementario que genere estrés, una vez que demanda la adquisición de conocimiento por parte de los alumnos, tanto para evaluaciones de enseñanza tradicional como para la búsqueda autónoma del conocimiento y las habilidades, que son las actitudes exigidas en la problematización. Objetivo Determinar el grado de resiliencia durante el curso de Medicina en un sistema híbrido de enseñanza, identificando mecanismos subyacentes. Métodos Estudio transversal desarrollado entre agosto del 2017 y agosto del 2018, en el centro universitario Christus, Brasil, una escuela de Medicina que utiliza currículos de aprendizaje con base en la problematización y asociados a la metodología tradicional de enseñanza. La escala de resiliencia de Wagnild y Young se aplicó a estudiantes de medicina de todos los semestres. También se compilaron variables socioeconómicas, emocionales e de desempeño académico, auto-referidas. La asociación entre las variables se evaluó con modelos de regresión logística mínimamente ajustados. Resultados 173 estudiantes de medicina participaron en este estudio, con un promedio de edad entre los 22,4 años, siendo que un 65,3% eran mujeres. El 88,1% de los estudiantes de Medicina presentaron alta o muy alta resiliencia. Recibir apoyo de los familiares y amigos, se asoció a una mejor resiliencia (valores de p inferiores a 0,001), los estudiantes que estaban "muy satisfechos" o "satisfechos" con el apoyo familiar tenían una tendencia mayor para desarrollar grados mejores de resiliencia, con resultados de tendencias de resiliencia muy elevadas (82,50%) y altas (71,10%), superando la prevalencia encontrada en estudiantes insatisfechos. Además, tener una creencia religiosa, también se asoció a grados más elevados de resiliencia (valor de p = 0,02). Conclusiones Los factores identificados en este estudio, principalmente la importancia de la red de apoyo de la familia y los amigos, pueden ser estimulados para mejorar la resiliencia de los alumnos. No se registró asociación directa entre la auto evaluación del desempeño académico y la resiliencia de los alumnos, siendo que ella tiende a permanecer constante a lo largo del curso.

Development and validation of rt-qpcr for vesicular stomatitis virus detection (Alagoas vesiculovirus).

Vesicular stomatitis is an infectious disease that occurs mainly in countries of the Western Hemisphere and affects cattle, swine and horses. The clinical symptoms in cattle and swine are similar to foot-and-mouth disease and include vesicular ulceration of the tongue and mouth. The disease requires a rapid and accurate differential diagnosis, aiming for immediate implementation of control measures. The objective of the present study was to develop and perform validation tests of multiplex RT-qPCR(s) for the detection of RNA from Alagoas vesiculovirus, considering the parameters of sensitivity and analytical specificity, analytical performance (repeatability and reproducibility criteria) and the uncertainty of the measurement. The threshold cycle values obtained in triplicate from each sample were evaluated by considering the variations between days, analysts and equipment in an analysis of variance aimed at determining the variances of repeatability and reproducibility. The results showed that RT-qPCRs had excellent sensitivity and specificity in the detection of RNA of the Alagoas vesiculovirus. The validation parameters showed low coefficients of variation and were equivalent to those found in other validation studies, indicating that the tests presented excellent repeatability and reproducibility.

Evaluation of PCSK9 levels and its genetic polymorphisms in women with polycystic ovary syndrome.

Dyslipidemia is one of the common metabolic disorders in Polycystic Ovary Syndrome (PCOS). Proprotein convertase subtilisin kexin type 9 (PCSK9) is a new component of lipid metabolism and correlated to the development of dyslipidemia and atherosclerosis. This protein acts by preventing the recycling of LDL receptors (LDL-r) back to the cell surface and thus generates higher levels of LDLc. The objective of this study was to evaluate the PCSK9 polymorphisms rs505151 (c.2009A>G), rs562556 (c.1420A>G) and rs11206510 (T>C) and plasma PCSK9 levels in PCOS. A group of women with PCOS (n=97), and a group of healthy women (control, n=99) were selected. Biochemical parameters were determined by using Vitros system and polymorphisms were assessed by TaqMan SNP Genotyping Assays. Plasma PCSK9 levels or PCSK9 polymorphisms were not associated with PCOS. The genotype rs11206510TT was associated with higher levels of PCSK9 in both groups. The population investigated (PCOS+control groups) with the rs505151AA genotype presented higher HDLc levels. The GG genotype regarding rs562556 polymorphism was associated with higher HDLc in PCOS group, while the AA genotype carriers had higher plasma testosterone levels when evaluated all women in a same group. The results were the same by comparing recessive and dominant model despite PCOS or both groups altogether. Our results suggest that PCSK9 is not altered specifically in PCOS, but it could be associated with in lipid and androgen metabolism in Brazilian women.

Metformin reduces total microparticles and microparticles-expressing tissue factor in women with polycystic ovary syndrome.

PURPOSE: The objective of this study was to evaluate the levels of total microparticles (MPs) and microparticles-expressing tissue factor (TFMPs) in women with polycystic ovarian syndrome (PCOS) who use metformin comparing to those who do not take metformin. METHODS: We quantified total MPs and TFMPs in the plasma of 50 patients with PCOS-13 of these women used metformin (850 mg 2×/day during at least 6 months) and the other 37 did not. For this purpose, the microparticles (MPs) were purified by differential centrifugation of the plasma and, subsequently, by flow cytometry, using annexin-V and CD142 as markers. RESULTS: Total MPs levels were lower in treated patients (59.58 ± 28.43 MPs/µL) when compared to untreated group (97.32 ± 59.42; p = 0.033). Plasma levels of TFMPs were also significantly lower in the group of patients who used metformin (1.10 ± 0.94 MPs/µL) when compared to untreated patients (2.20 ± 1.42 MPs/µL) (p = 0.003). CONCLUSIONS: Considering that metformin reduced the levels of total MPs and TFMPs, our results suggest that this mechanism could be involved in the antithrombotic metformin effect, corroborating with the indication of this drug in the PCOS treatment.

Peripheral blood-derived cytokine gene polymorphisms and metabolic profile in women with polycystic ovary syndrome.

BACKGROUND: The imbalance between proinflammatory and anti-inflammatory pathways plays a role in polycystic ovary syndrome (PCOS) etiology. We aimed to investigate the relationship between polymorphisms of genes encoding inflammation-associated cytokines and the metabolic profile of Brazilian women with PCOS. DESIGN: Case-control study. METHODS: The study included 196 women - 97 with PCOS (diagnosed based on Rotterdam criteria, 2003) and 99 age-matched, healthy women (controls). It was investigated polymorphisms in cytokines genes from peripheral blood-derived DNA by using PCR. RESULTS: The frequencies of alleles, genotypes, and phenotypes were similar between women with PCOS and controls. The GG genotype of the -179C/G polymorphism (IL6) was associated with higher glucose levels, while the GA and AA genotypes of the -1082A/G polymorphism (IL10), CT and TT genotypes of the -819A/T polymorphism (IL10), CA and AA genotypes of the -522A/G (IL10) polymorphism, and TA genotype of the +874T/A polymorphism (IFN-γ) were associated with lower total cholesterol and triglycerides levels. The GA genotype of the -1082A/G polymorphism (IL10) and the CC genotype of the 10T/C polymorphism (TGF-ß1) were associated with lower and higher Ferriman indices, respectively, in women with PCOS. The AA genotype of the -1082A/G polymorphism (IL10) was associated with lower glucose levels, while the TC genotype of the 10T/C polymorphism (TGF-ß1) was associated with a lower lipid accumulation product index and higher high-density lipoprotein cholesterol levels in the PCOS group. CONCLUSIONS: The genetic polymorphisms of cytokines are not associated with PCOS development, but may contribute to common metabolic disorders associated with PCOS.

Ferriman-Gallwey score correlates with obesity and insulin levels in polycystic ovary syndrome: an observational study / Correlação da escala de Ferriman-Gallwey com a obesidade e níveis de insulina na síndrome dos ovários policísticos: um estudo observacional

BACKGROUND: The Polycystic Ovary Syndrome (PCOS) is the most common endocrinopathy and one of the main causes of infertility in women. OBJECTIVES: This study aimed to evaluate the correlation between clinical hyperandrogenism assessed by modified Ferriman-Gallwey (F-G) score and metabolic parameters in Polycystic Ovary Syndrome women. METHODS: This observational study included fifty Polycystic Ovary Syndrome subjects. Detailed information about body mass index (BMI) and abdominal circumference (AC) were obtained from each subject. F-G score was applied to assess hirsutism through visual method. Serum levels of insulin, glucose and testosterone were measured. RESULTS: A positive correlation was observed between F-G score with body mass index, abdominal circumference and insulin. CONCLUSIONS: Obesity, mainly abdominal adipose tissue, and insulin levels correlate with hyperandrogenism in Polycystic Ovary Syndrome women, analyzed by F-G score. F-G score could be a marker to evaluate metabolic disorders in Polycystic Ovary Syndrome women.

JUSTIFICATIVA: A Síndrome dos Ovários Policísticos (SOP) é a endocrinopatia mais comum e uma das principais causas de infertilidade em mulheres. OBJETIVOS: O presente estudo teve como objetivo avaliar a correlação entre hiperandrogenismo clínico, avaliado pela escala de Ferriman-Gallwey (FG) modificada e parâmetros metabólicos em mulheres com a Síndrome dos Ovários Policísticos. MÉTODOS: Este estudo observacional incluiu cinquenta mulheres com Síndrome dos Ovários Policísticos. Informações detalhadas sobre o índice de massa corporal (IMC) e circunferência abdominal (CA) foram obtidas de cada participante. A escala FG foi aplicada para avaliar o hirsutismo através do método visual. Níveis séricos de insulina, glicose e testosterona foram também avaliados. RESULTADOS: Observou-se uma correlação positiva entre a escala FG e o índice de massa corporal, circunferência abdominal e insulina. CONCLUSÕES: A obesidade, principalmente o tecido adiposo abdominal, e os níveis de insulina se correlacionam com hiperandrogenismo em mulheres com Síndrome dos Ovários Policísticos, analisados por meio da escala FG. Desta forma, esta escala poderia ser um marcador para avaliar distúrbios metabólicos em mulheres com Síndrome dos Ovários Policísticos.

Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis

Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.

Evaluation of molecular markers for the diagnosis of Mycobacterium bovis.

Mycobacterium tuberculosis complex (MTC) comprises a group of bacteria that have a high degree of genetic similarity. Two species in this group, Mycobacterium tuberculosis and Mycobacterium bovis, are the main cause of human and bovine tuberculosis, respectively. M. bovis has a broader host range that includes humans; thus, the differentiation of mycobacterium is of great importance for epidemiological and public health considerations and to optimize treatment. The current study aimed to evaluate primers and molecular markers described in the literature to differentiate M. bovis and M. tuberculosis by PCR. Primers JB21/22, frequently cited in scientific literature, presented in our study the highest number of errors to identify M. bovis or M. tuberculosis (73%) and primers Mb.400, designed to flank region of difference 4 (RD4), were considered the most efficient (detected all M. bovis tested and did not detect any M. tuberculosis tested). Although also designed to flank RD4, primers Mb.115 misidentified eight samples due to primer design problems. The results showed that RD4 is the ideal region to differentiate M. bovis from other bacteria classified in MTC, but primer design should be considered carefully.

Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis.

Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.

Tuberculosis patients co-infected with Mycobacterium bovis and Mycobacterium tuberculosis in an urban area of Brazil.

In this cross-sectional study, mycobacteria specimens from 189 tuberculosis (TB) patients living in an urban area in Brazil were characterised from 2008-2010 using phenotypic and molecular speciation methods (pncA gene and oxyR pseudogene analysis). Of these samples, 174 isolates simultaneously grew on Löwenstein-Jensen (LJ) and Stonebrink (SB)-containing media and presented phenotypic and molecular profiles of Mycobacterium tuberculosis, whereas 12 had molecular profiles of M. tuberculosis based on the DNA analysis of formalin-fixed paraffin wax-embedded tissue samples (paraffin blocks). One patient produced two sputum isolates, the first of which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, and the second of which only grew on SB media and presented phenotypic profiles of Mycobacterium bovis. One patient provided a bronchial lavage isolate, which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, but had molecular profiles of M. bovis from paraffin block DNA analysis, and one sample had molecular profiles of M. tuberculosis and M. bovis identified from two distinct paraffin blocks. Moreover, we found a low prevalence (1.6%) of M. bovis among these isolates, which suggests that local health service procedures likely underestimate its real frequency and that it deserves more attention from public health officials.

Rifampicin-warfarin interaction leading to macroscopic hematuria: a case report and review of the literature.

BACKGROUND: Rifampicin remains one of the first-line drugs used in tuberculosis therapy. This drug's potential to induce the hepatic cytochrome P450 oxidative enzyme system increases the risk of drug-drug interactions. Thus, although the presence of comorbidities typically necessitates the use of multiple drugs, the co-administration of rifampicin and warfarin may lead to adverse drug events. We report a bleeding episode after termination of the co-administration of rifampicin and warfarin and detail the challenges related to international normalized ratio (INR) monitoring. CASE PRESENTATION: A 59-year-old Brazilian woman chronically treated with warfarin for atrial fibrillation (therapeutic INR range: 2.0-3.0) was referred to a multidisciplinary anticoagulation clinic at a university hospital. She showed anticoagulation resistance at the beginning of rifampicin therapy, as demonstrated by repeated subtherapeutic INR values. Three months of sequential increases in the warfarin dosage were necessary to reach a therapeutic INR, and frequent visits to the anticoagulation clinic were needed to educate the patient about her pharmacotherapy and to perform the warfarin dosage adjustments. The warfarin dosage also had to be doubled at the beginning of rifampicin therapy. However, four weeks after rifampicin discontinuation, an excessively high INR was observed (7.22), with three-day macroscopic hematuria and the need for an immediate reduction in the warfarin dosage. A therapeutic and stable INR was eventually attained at 50% of the warfarin dosage used by the patient during tuberculosis therapy. CONCLUSIONS: The present case exemplifies the influence of rifampicin therapy on warfarin dosage requirements and the increased risk of bleeding after rifampicin discontinuation. Additionally, this case highlights the need for warfarin weekly monitoring after stopping rifampicin until the maintenance dose of warfarin has decreased to the amount administered before rifampicin use. In particular, patients with cardiovascular diseases and active tuberculosis represent a group with a substantial risk of drug-drug interactions. Learning how to predict and monitor drug-drug interactions may help reduce the incidence of clinically significant adverse drug events.

Tuberculosis patients co-infected with Mycobacterium bovis and Mycobacterium tuberculosis in an urban area of Brazil

In this cross-sectional study, mycobacteria specimens from 189 tuberculosis (TB) patients living in an urban area in Brazil were characterised from 2008-2010 using phenotypic and molecular speciation methods (pncA gene and oxyR pseudogene analysis). Of these samples, 174 isolates simultaneously grew on Löwenstein-Jensen (LJ) and Stonebrink (SB)-containing media and presented phenotypic and molecular profiles of Mycobacterium tuberculosis, whereas 12 had molecular profiles of M. tuberculosis based on the DNA analysis of formalin-fixed paraffin wax-embedded tissue samples (paraffin blocks). One patient produced two sputum isolates, the first of which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, and the second of which only grew on SB media and presented phenotypic profiles of Mycobacterium bovis. One patient provided a bronchial lavage isolate, which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, but had molecular profiles of M. bovis from paraffin block DNA analysis, and one sample had molecular profiles of M. tuberculosis and M. bovis identified from two distinct paraffin blocks. Moreover, we found a low prevalence (1.6%) of M. bovis among these isolates, which suggests that local health service procedures likely underestimate its real frequency and that it deserves more attention from public health officials.