Prevalence of rhinitis and associated factors in adolescents and adults: a Global Asthma Network study.

OBJECTIVE: To determine the prevalence of allergic rhinitis and associated factors in adolescents and in their parents/guardians. METHODS: A cross-sectional study, applying a standardized and validated written questionnaire. Adolescents (13-14 years old; n=1,058) and their parents/guardians (mean age=42.1 years old; n=896) living in the city of Uruguaiana, southern Brazil, responded to the Global Asthma Network standard questionnaires. RESULTS: The prevalence of allergic rhinitis in adolescents was 28.0%, allergic rhinoconjunctivitis, 21.3%, and severe forms of allergic rhinitis, 7.8%. In the adults, the prevalence of allergic rhinitis was 31.7%. Some associated factors with allergic rhinitis in adolescents include low physical exercise (OR 2.16; 95%CI 1.15-4.05), having only one older sibling (OR 1.94; 95CI 1.01-3.72) and daily meat consumption (OR 7.43; 95% CI 1.53-36.11). In contrast, consuming sugar (OR 0.34; 95%CI 0.12-0.93) or olive oil (OR 0.33; 95%CI 0.13-0 .81) once or twice a week, and eating vegetables daily (OR 0.39; 95%CI 0.15-0.99) were considered factors negatively associated. In adults, exposure to fungi at home (OR 5.25; 95%CI 1.01-27.22) and consumption of meat once or twice a week (OR 46.45; 95CI 2.12-1020.71) were factors associated with the medical diagnosis of allergic rhinitis, while low education (OR 0.25; 95%CI 0.07-0.92) was found to be a factor negatively associated. CONCLUSIONS: The prevalence of allergic rhinitis in adolescents is high, as well as its medical diagnosis in adults living in Uruguaiana. Environmental factors, especially food habits, were associated with findings in both groups.

Prevalence of rhinitis and associated factors in adolescents and adults: a Global Asthma Network study / Prevalência de rinite e fatores associados em adolescentes e adultos: um estudo da Global Asthma Network

Abstract Objective: To determine the prevalence of allergic rhinitis and associated factors in adolescents and in their parents/guardians. Methods: A cross-sectional study, applying a standardized and validated written questionnaire. Adolescents (13-14 years old; n=1,058) and their parents/guardians (mean age=42.1 years old; n=896) living in the city of Uruguaiana, southern Brazil, responded to the Global Asthma Network standard questionnaires. Results: The prevalence of allergic rhinitis in adolescents was 28.0%, allergic rhinoconjunctivitis, 21.3%, and severe forms of allergic rhinitis, 7.8%. In the adults, the prevalence of allergic rhinitis was 31.7%. Some associated factors with allergic rhinitis in adolescents include low physical exercise (OR 2.16; 95%CI 1.15-4.05), having only one older sibling (OR 1.94; 95CI 1.01-3.72) and daily meat consumption (OR 7.43; 95% CI 1.53-36.11). In contrast, consuming sugar (OR 0.34; 95%CI 0.12-0.93) or olive oil (OR 0.33; 95%CI 0.13-0 .81) once or twice a week, and eating vegetables daily (OR 0.39; 95%CI 0.15-0.99) were considered factors negatively associated. In adults, exposure to fungi at home (OR 5.25; 95%CI 1.01-27.22) and consumption of meat once or twice a week (OR 46.45; 95CI 2.12-1020.71) were factors associated with the medical diagnosis of allergic rhinitis, while low education (OR 0.25; 95%CI 0.07-0.92) was found to be a factor negatively associated. Conclusions: The prevalence of allergic rhinitis in adolescents is high, as well as its medical diagnosis in adults living in Uruguaiana. Environmental factors, especially food habits, were associated with findings in both groups.

Resumo Objetivo: Determinar a prevalência de rinite alérgica e fatores associados em adolescentes e em seus pais/responsáveis. Métodos: Estudo transversal, com aplicação de questionário escrito padronizado e validado. Adolescentes (13-14 anos; n=1.058) e seus pais/responsáveis (média de idade=42,1 anos; n=896) residentes na cidade de Uruguaiana, Sul do Brasil, responderam aos questionários padrão da Global Asthma Network. Resultados: A prevalência de rinite alérgica em adolescentes foi de 28,0%, a de rinoconjuntivite alérgica foi de 21,3% e a de formas graves de rinite alérgica, de 7,8%. Nos adultos, a prevalência de rinite alérgica foi de 31,7%. Alguns fatores associados à rinite alérgica em adolescentes incluem fazer pouco exercício físico (odds ratio — OR 2,16; intervalo de confiança — IC95% 1,15-4,05), ter apenas um irmão mais velho (OR 1,94; IC95% 1,01-3,72) e consumir carne diariamente (OR 7,43; IC95% 1,53-36,11). Por outro lado, consumir açúcar (OR 0,34; IC5% 0,12-0,93) ou azeite de oliva (OR 0,33 IC95% 0,13-0,81) uma ou duas vezes por semana e comer vegetais diariamente (OR 0,39; IC95% 0,15-0,99) foram considerados fatores associados negativamente. Em adultos, a exposição a fungos no domicílio (OR 5,25; IC95% 1,01-27,22) e o consumo de carne uma ou duas vezes por semana (OR 46,45; IC95 2,12-1020,71) foram fatores associados ao diagnóstico médico de rinite alérgica, enquanto a baixa escolaridade (OR 0,25; IC95% 0,07-0,92) se mostrou como fator associado negativamente. Conclusões: A prevalência de rinite alérgica em adolescentes é alta, assim como seu diagnóstico médico em adultos residentes em Uruguaiana. Fatores ambientais, especialmente hábitos alimentares, foram associados aos achados em ambos os grupos.

Prevalence and factors associated with depression and anxiety among medical students in an inland university in Brazil / Prevalência e fatores associados à depressão e ansiedade entre estudantes de medicina de uma universidade do interior do Brasil

Medical schools are known to be stressful environments for students, and hence medical students have been believed to experience greater incidences of depression and anxiety than the general population or students from other spe-cialties. The present study investigates the prevalence of anxiety and depressive symptoms together with factors as-sociated with them among medical students in a federal public university in the south of Brazil. A descriptive study was performed using self-administered questionnaires to access socio-demographic, institutional, and health variables in association with two scales - Beck's Depression Inventory (BDI) and State-Trait Anxiety Inventory (STAI) - designed to assess depressive and anxiety symptoms, respectively. The research sample consisted of 152 subjects. The depres-sive symptoms prevalence was 65.1% (BDI > 9), state-anxiety was 98.6%, and trait-anxiety was 97.4% (STAI > 33). Among women, 37.8% demonstrated moderate/severe depressive symptoms. High levels of state-anxiety symptoms and trait-anxiety symptoms were found in 44.7% of students under the age of 20. In the group with the lowest month-ly income, it was observed the highest distributions for moderate/severe depressive symptoms, high state and high trait-anxiety symptoms, corresponding to 47.4%, 57.9%, and 47.4%, respectively. Students attending the third term of medical school had the highest percentage of moderate/severe depressive symptoms (62.5%) and high state-anxiety symptoms (50%). In addition, students who had both financial aid programs presented the highest percentages of moderate/severe depressive symptoms (46.2%), highstate-anxiety symptoms (61.5%), and high trait-anxiety symptoms (46.2%). We also observed a correlationbetween depression and trait-anxiety symptoms (P = 0.037). In conclusion, it was identified as risk factors for depressive symptoms the previous depression diagnosis, previous search for health service due to psychological symptoms, being in financial aid programs, dissatisfaction with the medical school, and inadequate psychological help offered by it. For state-anxiety symptoms and trait-anxiety outcome, there is an increased risk among low-income or students who have financial help from financial aid programs and younger age students. (AU)

Objetivo: Investigar a prevalência de sintomas de ansiedade e depressão e fatores associados a eles entre es-tudantes de medicina de uma universidade pública do sul do Brasil. Métodos: Foi realizado um estudo descritivo através de questionários autoaplicáveis para avaliar variáveis sociodemográficas, institucionais e de saúde, associa-das a duas escalas - Inventário de Depressão de Beck (BDI) e Inventário de Traço-Estado de Ansiedade (STAI) - para avaliar os sintomas de depressão e ansiedade, respectivamente. Resultados: A prevalência de sintomas depres-sivos foi de 65,1% (BDI > 9), estado de ansiedade foi de 98,6% e traço de ansiedade de 97,4% (STAI > 33). Uma correlação significativa foi encontrada entre depressão e traço de ansiedade. Diagnóstico prévio de depressão, busca prévia por serviços de saúde, insatisfação com o curso de medicina e ter auxílio de baixa renda e bolsa de iniciação científica foram identificados como fatores de risco para depressão. Estudantes com baixa renda mensal e menores de 25 anos apresentaram maior risco para estado de ansiedade. Idade, ano da faculdade de medicina e programa de ajuda financeira provaram ser um fator de risco para traço de ansiedade. Conclusão: Esses resultados mostram fatores significativos relacionados à saúde mental de estudantes de uma faculdade de medicina recém-fundada no interior do Brasil. (AU)

Prevalence of asthma symptoms and associated factors in adolescents and adults in southern Brazil: A Global Asthma Network Phase I study.

BACKGROUND: The Global Asthma Network (GAN) aims to find out the current status of the prevalence and severity of asthma, rhinitis, and eczema using global surveillance to achieve worldwide recognition and improve the management of asthma, especially in low- and middle-income countries. The aim of this study was to verify the associated factors for asthma in adolescents and their respective parents/caregivers. METHODS: Adolescents (13-14 years old; n = 1058) and their respective parents/caregivers (mean age = 42.1 years, n = 896) living in the town of Uruguaiana, Southern Brazil fulfilled the standardized questionnaire. RESULTS: Although the prevalence of wheezing in the past 12 months was higher among adults than adolescents (18.4% vs. 15.8%, respectively), adolescents showed more severe wheezing and worse control over the disease revealed by higher consumption of short-acting beta-2 agonists; going to the emergency room; hospitalization in the last year and dry night cough. Smoking and paracetamol use were associated with risk for developing asthma symptoms and consuming seafood/fish was protective. For the adults smoking (10 or more cigarettes/day) and exposure to mould in the house were associated with risk for asthma symptoms. CONCLUSIONS: Adolescents have a high prevalence of asthma symptoms and few have an action plan. Adults do not have their disease under control and they use more relief than preventive medication. Differences in associated factors could determine the outcomes in asthma control among adolescents and their parents.

Prevalência e fatores associados a sintomas de eczema atópico em adolescentes e adultos residentes na Região Sul do Brasil ­ Resultados do Global Asthma Network (GAN) / Prevalence and factors associated with symptoms of atopic eczema in adolescents and adults living in southern Brazil - Global Asthma Network (GAN) results

Objetivo: Embora o objetivo da Rede Global de Asma (GAN) seja entender o estado atual do eczema atópico (EA), sua prevenção e melhoria geral em seu manejo, particularmente em países de baixa e média renda, ela permite a avaliação de outras doenças alérgicas, como a asma (A) e a rinite alérgica (RA). Nosso objetivo foi determinar a prevalência de EA e fatores associados em adolescentes e seus pais/responsáveis. Método: Adolescentes (13-14 anos; n = 1.058) e seus pais/responsáveis (média = 42,1 anos; n = 896) residentes na cidade de Uruguaiana, RS, sul do Brasil, responderam aos questionários padrão do GAN. Resultados: A prevalência de EA em adolescentes foi de 8%, e a de formas graves foi de 1,3%, com predomínio no sexo feminino (67,8%). Nos adultos, a prevalência de EA foi de 3,1%. Alguns fatores de risco associados ao EA em adolescentes incluem o consumo de azeite ou de margarina. Em adultos, a exposição à umidade e manchas no passado e atual, uso de cigarros eletrônicos/narguilé, e consumo de outros laticínios foram associados a risco, e o consumo de arroz a proteção. Conclusões: A prevalência de EA em adolescentes é alta e predomina em mulheres, assim como o diagnóstico médico de EA em adultos residentes em Uruguaiana. Fatores ambientais, especialmente hábitos alimentares e umidade no domicílio, foram associados aos achados em ambos os grupos.

Objective: Although the purpose of the Global Asthma Network (GAN) is to understand the current status of atopic eczema (AE), including prevention and general improvement in its management, particularly in low- and middle-income countries, it allows the assessment of other allergic diseases such as asthma and allergic rhinitis. Our objective was to determine the prevalence of AE and associated factors in adolescents and their parents/guardians. Method: Adolescents (13-14 years old; n = 1058) and their parents/guardians (mean = 42.1 years old; n = 896) living in the city of Uruguaiana, RS, southern Brazil, completed the standard GAN questionnaires. Results: The prevalence of AE in adolescents was 8%, and that of severe forms was 1.3%, with predominance in females (67.8%). In adults, the prevalence of AE was 3.1%. Some risk factors associated with AE in adolescents include the consumption of oil or margarine. In adults, exposure to moisture and stains in the past and currently, use of electronic cigarettes/ hookahs, and consumption of other dairy products were identified as risk factors, while rice consumption was considered a protective factor. Conclusions: The prevalence of AE in adolescents is high and the disease predominates in women, as well as the medical diagnosis of AE in adults living in Uruguaiana. Environmental factors, especially eating habits and moisture at home, were associated with the findings in both groups.

Poluição atmosférica: o cigarro nosso de todo o dia / Air pollution: our daily cigarette

O objetivo deste artigo foi avaliar a relação entre poluição atmosférica e a saúde das crianças. Foi realizada pesquisa na base de dados do MEDLINE, nos últimos 10 anos, empregando os seguintes termos: "air pollution", ou "household air pollution" ou "primary traffic pollutants" ou "environmental burden" AND "child health" ou "lung function" ou "acute respiratory infections" ou "pregnancy". Os artigos identificados foram selecionados após leitura dos títulos, e os considerados de importância foram lidos na íntegra e incluídos nessa revisão. Além desses, referências por eles citadas e dados de agências oficiais, considerados de relevância, também foram incluídos. Estudos documentam associação positiva entre exposição de crianças à poluição do ar e maior morbimortalidade de afecções respiratórias, incluindo a asma. Quanto mais precoce for essa exposição, maiores serão os seus efeitos.

The objective of this study was to evaluate the relationship between air pollution and children's health. A search was performed in the MEDLINE database for the past 10 years using the following terms: "air pollution" or "household air pollution" or "primary traffic pollutants" AND "child health" or "lung function" or "acute respiratory infections" or "pregnancy". For selection of articles, titles were read and analyzed, and articles considered of importance were read in full and then included in this review. In addition, cited references and relevant data from official agencies were included. The selected studies report a positive association between exposure of children to air pollution and increased morbidity and mortality due to respiratory diseases, including asthma. The earlier the exposure, the greater its effects.

Conhecimento de farmacêuticos sobre rinite alérgica e seu impacto na asma (guia ARIA para farmacêuticos): um estudo piloto comparativo entre Brasil e Paraguai / Knowledge of allergic rhinitis and its impact on asthma among pharmacists (ARIA guidelines for pharmacists): a comparative pilot study in Brazil and Paraguay

Introdução: Os farmacêuticos, geralmente, são os primeiros profissionais a atenderem pacientes com rinite alérgica (RA). A guia ARIA (Rinite Alérgica e seu Impacto na Asma) estabelece padrões de melhores práticas para o manejo de pacientes com RA. Objetivo: Avaliar o nível de conhecimento sobre RA e a guia ARIA entre farmacêuticos do Brasil (BR) e Paraguai (PY). Método: 205 farmacêuticos (BR = 78, PY = 127) responderam ao questionário online autoaplicável (ARIA One Airways) sobre dados pessoais, profissionais e conhecimento sobre RA e guia ARIA, empregando o Google Forms. Resultados: A mediana de idade foi 32 anos, 35% BR e 52% PY referiam terem tido mais de quatro anos de treinamento. Embora reconheçam os principais sintomas de RA, 26% BR e 100% PY nunca perguntaram se o paciente tinha diagnóstico médico de RA; 20,5% BR e 100,0% PY se os sintomas ocorreram quando perto de animais ou alérgenos; 55% BR e 76% PY se o paciente tinha diagnóstico médico de asma; 59% BR e 70% PY se a rinite piora os sintomas de asma. Anti-histamínicos sedantes foram recomendados por 34,6% BR e 26,8% PY, e corticosteroides intranasais por 59% BR e 52% PY. Embora 85% BR e 100% PY desconheçam as diretrizes ARIA, 94,9% BR e 60,6% PY encaminhariam o paciente a um especialista. Conclusão: Embora os farmacêuticos sejam os primeiros profissionais procurados pelo paciente com RA para alívio de sintomas, o seu nível de conhecimento sobre RA e a guia ARIA é baixo. O seu treinamento para atingir melhor abordagem clínica é primordial.

Introduction: Pharmacists are usually the first professionals to see patients with allergic rhinitis (AR). The Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines establish best practice standards for the management of patients with AR. Objective: To evaluate the level of knowledge of AR and ARIA guideline recommendations among pharmacists in Brazil (BR) and Paraguay (PY). Method: A total of 205 pharmacists (BR = 78, PY = 127) answered a self-administered online questionnaire (ARIA One Airways), using the Google Forms tool, containing questions covering personal data, professional data, and knowledge of AR and ARIA guidelines. Results: Median age was 32 years; 35% BR and 52% PY reported having had more than four years of training. Although they recognized the main symptoms of AR, 26% BR and 100% PY never asked whether the patient had a medical diagnosis of AR; 20.5% BR and 100.0% PY if symptoms occurred when close to animals or allergens; 55% BR and 76% PY if the patient had a medical diagnosis of asthma; 59% BR and 70% PY if rhinitis worsened asthma symptoms. Sedating antihistamines were recommended by 34.6% BR and 26.8% PY, and intranasal corticosteroids by 59% BR and 52% PY. Although 85% BR and 100% PY were unaware of ARIA guidelines, 94.9% BR and 60.6% PY would refer the patient to a specialist. Conclusion: Even though pharmacists are the first professionals sought by patients with AR for symptom relief, their level of knowledge of AR and ARIA guidelines is very low. Training is paramount to improve clinical management of AR among pharmacists.

[Primary care physicians' knowledge about allergic rhinitis and its impact on asthma (ARIA guidelines): a comparative Brazilian/Paraguayan/Uruguayan pilot study]. / Conocimiento de médicos de atención primaria sobre Rinitis Alérgica y su Impacto en el Asma (ARIA): estudio piloto comparativo Brasil, Paraguay, Uruguay.

BACKGROUND: Allergic Rhinitis and its Impact on Asthma (ARIA) establishes guidelines for the treatment of allergic rhinitis (AR) and its comorbidities. OBJECTIVE: To assess and compare the level of knowledge about AR and ARIA guidelines among primary care physicians (PCP) of Brazil, Paraguay and Uruguay. METHODS: 336 PCPs (Brazil 109, Paraguay 127, Uruguay 100) answered a questionnaire about personal and professional data and knowledge on RA and the ARIA guidelines; the Google Forms tool was used. RESULTS: There was a predominance of females (73, 50.4 and 70 % in Brazil, Paraguay and Uruguay, respectively); 124 worked in the community and 75 in emergency departments. Brazil and Uruguay PCPs recognized AR symptoms; 67 % of those from Uruguay did not ask about a history of AR diagnosis, 93 % about an asthma diagnosis and 90 % about rhinitis interference with daily activities. Intranasal corticosteroids use: 78, 92 and 54 % of Brazilian, Paraguayan and Uruguayan doctors; 100, 73.4 and 78 % of Paraguayan, Brazilian and Uruguayan PCPs, respectively, did not refer the patient to a specialist; 26.6, 62 and 6 % of PCPs in Brazil, Paraguay and Uruguay, respectively, knew the ARIA guidelines. CONCLUSIONS: AR is commonly looked after by PCPs, but the recognition of symptoms and knowledge about ARIA guidelines are low and treatment is not always optimal.

Antecedentes: En ARIA se establecen directrices para el tratamiento de la rinitis alérgica y sus comorbilidades. Objetivo: Evaluar y comparar el nivel de conocimiento sobre rinitis alérgica y la guía ARIA en médicos de atención primaria (MAP) de Brasil, Paraguay y Uruguay. Métodos: 336 MAP (Brasil 109, Paraguay 127, Uruguay 100) respondieron un cuestionario sobre datos personales, profesionales y conocimiento en rinitis alérgica y ARIA; se utilizó Google Forms. Resultados: Predominaron las mujeres (73, 50.4 y 70 % en Brasil Paraguay Uruguay); 124 trabajaban en la comunidad y 75 en servicios de urgencias. Los MAP de Brasil y Uruguay reconocieron los síntomas de rinitis alérgica; 67 % de Uruguay no preguntaba sobre antecedentes de diagnóstico de rinitis alérgica, 93 % sobre diagnóstico de asma y 90 % sobre la interferencia de la rinitis con las actividades diarias. Uso de corticosteroides intranasales: 78, 92 y 54 % de los médicos de Brasil Paraguay y Uruguay; 100, 73.4 y 78 % de Paraguay, Brasil y Uruguay no derivaban el paciente al especialista; 26.6, 62 y 6 % de Brasil, Paraguay y Uruguay conocían ARIA. Conclusiones: La rinitis alérgica frecuentemente es atendida por el MAP, pero el reconocimiento de los síntomas y el conocimiento sobre ARIA son reducidos y el tratamiento no siempre es óptimo.

Alginate-embedded HuH-7 cells increase MMP-9 and reduce OCLN expression in vitro.

BACKGROUND: Hepatocellular carcinoma is a common cancer, ranking third in cancer-associated deaths. An important cause of cancer patients' mortality is metastasis. At the start of metastasis progression, there is an epithelial-mesenchymal transition, characterized by matrix degradation, junction reductions and vessels formation. HuH-7 is a cell line used in research as an in vitro model for hepatocellular carcinoma. It is known that two-dimensional growth reflects tumor characteristics poorly. In contrast, three-dimensional cultures provide a better approach to the study of tumorigenic potential. The purpose of this work was to mimic a three-dimensional environment in order to assess gene expression of some epithelial-mesenchymal transition and metastasis progression markers in HuH-7 cells and compare them with traditional two-dimensional culture model. METHODS: HuH-7 cells were encapsulated in sodium alginate (three-dimensional model) to be compared with cells grown in two-dimensional flasks. After 4 days in culture, gene expression of Matrix metallopeptidase 9, Occludin, p65, Intercellular adhesion molecule 1 and Vascular endothelial growth factor A was analyzed by qPCR and cytoskeleton assessment was performed by rhodamine-phalloidin staining. RESULTS: Differences were found in gene expression, with a high increment of Matrix metallopeptidase 9 and Occludin reduction. The cytoskeleton morphology also showed differences, with a cytoplasm restricted only near the nuclei in the three-dimensional model. CONCLUSIONS: This work shows the effects of using sodium alginate capsules as a three-dimensional model to the study of HuH-7. Cells in this 3D system show key markers of epithelial-mesenchymal transition, such as Matrix metallopeptidase 9 overexpression and Occludin down-regulation.

Prevalence and Phenotypic Expression of Mutations in the MYH7, MYBPC3 and TNNT2 Genes in Families with Hypertrophic Cardiomyopathy in the South of Brazil: A Cross-Sectional Study.

BACKGROUND:: Mutations in sarcomeric genes are found in 60-70% of individuals with familial forms of hypertrophic cardiomyopathy (HCM). However, this estimate refers to northern hemisphere populations. The molecular-genetic profile of HCM has been subject of few investigations in Brazil, particularly in the south of the country. OBJECTIVE:: To investigate mutations in the sarcomeric genes MYH7, MYBPC3 and TNNT2 in a cohort of HCM patients living in the extreme south of Brazil, and to evaluate genotype-phenotype associations. METHODS:: Direct DNA sequencing of all encoding regions of three sarcomeric genes was conducted in 43 consecutive individuals of ten unrelated families. RESULTS:: Mutations for CMH have been found in 25 (58%) patients of seven (70%) of the ten study families. Fourteen (56%) individuals were phenotype-positive. All mutations were missense, four (66%) in MYH7 and two (33%) in MYBPC3. We have not found mutations in the TNNT2 gene. Mutations in MYH7 were identified in 20 (47%) patients of six (60%) families. Two of them had not been previously described. Mutations in MYBPC3 were found in seven (16%) members of two (20%) families. Two (5%) patients showed double heterozygosis for both genes. The mutations affected different domains of encoded proteins and led to variable phenotypic expression. A family history of HCM was identified in all genotype-positive individuals. CONCLUSIONS:: In this first genetic-molecular analysis carried out in the south of Brazil, we found mutations in the sarcomeric genes MYH7 and MYBPC3 in 58% of individuals. MYH7-related disease was identified in the majority of cases with mutation. FUNDAMENTO:: Mutações em genes do sarcômero são encontradas em 60-70% dos indivíduos com formas familiares de cardiomiopatia hipertrófica. (CMH). Entretanto, essa estimativa refere-se a populações de países do hemisfério norte. O perfil genético-molecular da CMH foi tema de poucos estudos no Brasil, particularmente na região sul do país. OBJETIVO:: Realizar a pesquisa de mutações dos genes sarcoméricos MYH7, MYBPC3 e TNNT2 numa coorte de CMH estabelecida no extremo sul do Brasil, assim como avaliar as associações genótipo-fenótipo. MÉTODOS:: Sequenciamento direto do DNA de todas as regiões codificantes dos três genes sarcoméricos foi realizada em 43 indivíduos consecutivos de dez famílias não-relacionadas. RESULTADOS:: Mutações para CMH foram encontradas em 25 (58%) indivíduos de sete (70%) das dez famílias estudadas, sendo 14 (56%) deles fenótipo-positivos. Todas as mutações eram missense, quatro (66%) no gene MYH7 e duas (33%) no gene MYBPC3. Não foram encontradas mutações no gene TNNT2. Mutações em MYH7 foram identificadas em 20 (47%) indivíduos de seis (60%) famílias. Duas delas não haviam sido previamente relatadas. Mutações de MYBPC3 foram detectadas em sete (16%) membros de duas (20%) famílias. Dois (5%) indivíduos apresentaram dupla heterozigose com mutações em ambos os genes. As mutações acometeram distintos domínios das proteínas codificadas e produziram expressão fenotípica variável. História familiar de CMH foi identificada em todos os indivíduos genótipo-positivos. CONCLUSÕES:: Nessa primeira análise genético-molecular da CMH realizada no sul do Brasil, foram encontradas mutações nos genes sarcoméricos MYH7 e MYBPC3 em 58% dos indivíduos. Doença relacionada ao gene MYH7 foi identificada na maioria dos casos com mutação.

Prevalence and Phenotypic Expression of Mutations in the MYH7, MYBPC3 and TNNT2 Genes in Families with Hypertrophic Cardiomyopathy in the South of Brazil: A Cross-Sectional Study / Prevalência e Expressão Fenotípica das Mutações nos Genes MYH7, MYBPC3 e TNNT2 em Famílias com Cardiomiopatia Hipertrófica no Sul do Brasil: Um Estudo Transversal

Abstract Background: Mutations in sarcomeric genes are found in 60-70% of individuals with familial forms of hypertrophic cardiomyopathy (HCM). However, this estimate refers to northern hemisphere populations. The molecular-genetic profile of HCM has been subject of few investigations in Brazil, particularly in the south of the country. Objective: To investigate mutations in the sarcomeric genes MYH7, MYBPC3 and TNNT2 in a cohort of HCM patients living in the extreme south of Brazil, and to evaluate genotype-phenotype associations. Methods: Direct DNA sequencing of all encoding regions of three sarcomeric genes was conducted in 43 consecutive individuals of ten unrelated families. Results: Mutations for CMH have been found in 25 (58%) patients of seven (70%) of the ten study families. Fourteen (56%) individuals were phenotype-positive. All mutations were missense, four (66%) in MYH7 and two (33%) in MYBPC3. We have not found mutations in the TNNT2 gene. Mutations in MYH7 were identified in 20 (47%) patients of six (60%) families. Two of them had not been previously described. Mutations in MYBPC3 were found in seven (16%) members of two (20%) families. Two (5%) patients showed double heterozygosis for both genes. The mutations affected different domains of encoded proteins and led to variable phenotypic expression. A family history of HCM was identified in all genotype-positive individuals. Conclusions: In this first genetic-molecular analysis carried out in the south of Brazil, we found mutations in the sarcomeric genes MYH7 and MYBPC3 in 58% of individuals. MYH7-related disease was identified in the majority of cases with mutation.

Resumo Fundamento: Mutações em genes do sarcômero são encontradas em 60-70% dos indivíduos com formas familiares de cardiomiopatia hipertrófica. (CMH). Entretanto, essa estimativa refere-se a populações de países do hemisfério norte. O perfil genético-molecular da CMH foi tema de poucos estudos no Brasil, particularmente na região sul do país. Objetivo: Realizar a pesquisa de mutações dos genes sarcoméricos MYH7, MYBPC3 e TNNT2 numa coorte de CMH estabelecida no extremo sul do Brasil, assim como avaliar as associações genótipo-fenótipo. Métodos: Sequenciamento direto do DNA de todas as regiões codificantes dos três genes sarcoméricos foi realizada em 43 indivíduos consecutivos de dez famílias não-relacionadas. Resultados: Mutações para CMH foram encontradas em 25 (58%) indivíduos de sete (70%) das dez famílias estudadas, sendo 14 (56%) deles fenótipo-positivos. Todas as mutações eram missense, quatro (66%) no gene MYH7 e duas (33%) no gene MYBPC3. Não foram encontradas mutações no gene TNNT2. Mutações em MYH7 foram identificadas em 20 (47%) indivíduos de seis (60%) famílias. Duas delas não haviam sido previamente relatadas. Mutações de MYBPC3 foram detectadas em sete (16%) membros de duas (20%) famílias. Dois (5%) indivíduos apresentaram dupla heterozigose com mutações em ambos os genes. As mutações acometeram distintos domínios das proteínas codificadas e produziram expressão fenotípica variável. História familiar de CMH foi identificada em todos os indivíduos genótipo-positivos. Conclusões: Nessa primeira análise genético-molecular da CMH realizada no sul do Brasil, foram encontradas mutações nos genes sarcoméricos MYH7 e MYBPC3 em 58% dos indivíduos. Doença relacionada ao gene MYH7 foi identificada na maioria dos casos com mutação.

Encapsulated platelets modulate kupffer cell activation and reduce oxidative stress in a model of acute liver failure.

Acute liver failure (ALF) is characterized by massive hepatocyte cell death. Kupffer cells (KC) are the first cells to be activated after liver injury. They secrete cytokines and produce reactive oxygen species, leading to apoptosis of hepatocytes. In a previous study, we showed that encapsulated platelets (PLTs) increase survival in a model of ALF. Here, we investigate how PLTs exert their beneficial effect. Wistar rats submitted to 90% hepatectomy were treated with PLTs encapsulated in sodium alginate or empty capsules. Animals were euthanized at 6, 12, 24, 48, and 72 hours after hepatectomy, and livers were collected to assess oxidative stress, caspase activity, and gene expression related to oxidative stress or liver function. The number of KCs in the remnant liver was evaluated. Interaction of encapsulated PLTs and KCs was investigated using a coculture system. PLTs increase superoxide dismutase and catalase activity and reduce lipid peroxidation. In addition, caspase 3 activity was reduced in animals receiving encapsulated PLTs at 48 and 72 hours. Gene expression of endothelial nitric oxide synthase and nuclear factor kappa B were elevated in the PLT group at each time point analyzed. Gene expression of albumin and factor V also increased in the PLT group. The number of KCs in the PLT group returned to normal levels at 12 hours but remained elevated in the control group until 72 hours. Finally, PLTs modulate interleukin (IL) 6 and IL10 expression in KCs after 24 hours of coculture. In conclusion, these results indicate that PLTs interact with KCs in this model and exert their beneficial effect through reduction of oxidative stress that results in healthier hepatocytes and decreased apoptosis. Liver Transplantation 22 1562-1572 2016 AASLD.

Injured hepatocyte-released microvesicles induce bone marrow-derived mononuclear cells differentiation.

The ability of bone marrow-derived mononuclear cells (BMMCs) to differentiate into hepatocyte-like cells under different conditions has been demonstrated previously. In the present study, we investigated the effect of CCl4-injured hepatocytes on the differentiation of the non-adherent (NAD) fraction of BMMCs. Differentiation (cell fate) was analyzed after 2, 6 and 24h of co-culture by gene and protein expression and by urea production. We also evaluated the presence of microvesicles (MVs) in the supernatant of differentiated cells, their content and the ability of these cells to absorb them. Hepatocyte-like characteristics were observed in the NAD cells after 24h of co-culture with injured hepatocytes. Cells that were co-cultured with healthy hepatocytes did not present signs of differentiation at any analyzed time point. Analysis of the supernatant from differentiated cells revealed the presence of MVs carrying hepatocyte-specific mRNAs, including Albumin, Coagulation factor V, Alpha-fetoprotein, and Cytokeratin 18. The incorporation of injured hepatocyte-derived MVs by NAD cells was shown at 24h, suggesting a possible role for MVs in the induction of cell plasticity.

Fabry disease: a new approach for the screening of females in high-risk groups.

OBJECTIVE: Fabry disease (FD) is a rare X-linked inborn error of metabolism caused by deficient activity of lysosomal α-galactosidase A (α-GAL). Due to random X inactivation, α-GAL activity in heterozygous females ranges from very low to overlapping normal values. Determining this specific range and altering assays cutoffs could become a valuable tool for minimizing the need in DNA sequencing for screening of all potential carriers. Therefore, the aim of this study was to establish the range of enzyme in dried blood spots (DBS), plasma and leukocytes that suggests carrier status for FD. DESIGN AND METHODS: α-GAL gene was sequenced in 453 women with clinical suspicion and/or positive family history of FD. This data was compared to the α-GAL activity measured in DBS (dried blood spots) and/or plasma and/or leukocytes. RESULTS: About 12% of the samples had pathogenic mutations (c.30_32delG, c.718_719delAA, p.R118C, p.S126G, p.Y152X, p.A156D, p.C202Y, p.N215S, p.P259R, p.D264Y, p.V269M, p.R342Q and p.R356W). When compared to genotype, DBS was the least reliable biochemical test for screening, with very low specificity. Plasma and leukocyte activities presented high AUC in ROC curve analysis, both over 84%. When cutoffs were altered to identify all carriers, leukocyte specificity was higher than that of plasma (35.2% and 27.6%, respectively). Moderated correlation and agreement coefficients were found between them, which reinforces the need for using both data combined. CONCLUSION: A combined approach involving plasma and leukocyte α-GAL activities, with distinct cutoffs for men and women, could represent a more accurate, faster and less expensive tool to screen women for FD in high-risk groups in middle- and low-income countries.

Diagnostic contribution of molecular analysis of the cystic fibrosis transmembrane conductance regulator gene in patients suspected of having mild or atypical cystic fibrosis.

OBJECTIVE: To evaluate the diagnostic contribution of molecular analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in patients suspected of having mild or atypical cystic fibrosis (CF). METHODS: This was a cross-sectional study involving adolescents and adults aged > 14 years. Volunteers underwent clinical, laboratory, and radiological evaluation, as well as spirometry, sputum microbiology, liver ultrasound, sweat tests, and molecular analysis of the CFTR gene. We then divided the patients into three groups by the number of mutations identified (none, one, and two or more) and compared those groups in terms of their characteristics. RESULTS: We evaluated 37 patients with phenotypic findings of CF, with or without sweat test confirmation. The mean age of the patients was 32.5 ± 13.6 years, and females predominated (75.7%). The molecular analysis contributed to the definitive diagnosis of CF in 3 patients (8.1%), all of whom had at least two mutations. There were 7 patients (18.9%) with only one mutation and 26 patients (70.3%) with no mutations. None of the clinical characteristics evaluated was found to be associated with the genetic diagnosis. The most common mutation was p.F508del, which was found in 5 patients. The combination of p.V232D and p.F508del was found in 2 patients. Other mutations identified were p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P, p.R1066H, and p.T351S. CONCLUSIONS: The molecular analysis of the CFTR gene coding region showed a limited contribution to the diagnostic investigation of patients suspected of having mild or atypical CF. In addition, there were no associations between the clinical characteristics and the genetic diagnosis.

Contribuição da análise molecular do gene regulador da condutância transmembrana na fibrose cística na investigação diagnóstica de pacientes com suspeita de fibrose cística leve ou doença atípica / Diagnostic contribution of molecular analysis of the cystic fibrosis transmembrane conductance regulator gene in patients suspected of having mild or atypical cystic fibrosis

OBJETIVO: Avaliar a contribuição da análise molecular do gene cystic fibrosis transmembrane conductance regulator (CFTR, regulador da condutância transmembrana na fibrose cística) na investigação diagnóstica da fibrose cística em pacientes com suspeita de fibrose cística (FC) leve ou atípica. Métodos: Estudo transversal em adolescentes e adultos (idade > 14 anos). Os voluntários foram submetidos à avaliação clínica, laboratorial e radiológica; espirometria, microbiologia do escarro, ecografia hepática, teste do suor e análise molecular do gene CFTR. Compararam-se as características dos pacientes divididos em três grupos, segundo o número de mutações identificadas (duas ou mais, uma e nenhuma). Resultados: Foram avaliados 37 pacientes com achados fenotípicos de FC, com ou sem confirmação pelo teste do suor. Houve predomínio do sexo feminino (75,7%), e a média de idade dos participantes foi de 32,5 ± 13,6 anos. A análise molecular contribuiu para o diagnóstico de FC em 3 casos (8,1%), todos esses com pelo menos duas mutações. Houve a identificação de uma e nenhuma mutação, respectivamente, em 7 (18,9%) e 26 pacientes (70,3%). Nenhuma característica clínica estudada se associou com o diagnóstico genético. A mutação p.F508del foi a mais comum, encontrada em 5 pacientes. A associação de p.V232D e p.F508del foi encontrada em 2 pacientes. Outras mutações encontradas foram p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P, p.R1066H e p.T351S. Conclusões: A análise molecular da região codificadora do gene CFTR apresentou uma contribuição limitada para a investigação diagnóstica desses pacientes com suspeita de FC leve ou atípica. Além disso, não houve associações entre as características clínicas e o diagnóstico genético.

OBJECTIVE: To evaluate the diagnostic contribution of molecular analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in patients suspected of having mild or atypical cystic fibrosis (CF). METHODS: This was a cross-sectional study involving adolescents and adults aged > 14 years. Volunteers underwent clinical, laboratory, and radiological evaluation, as well as spirometry, sputum microbiology, liver ultrasound, sweat tests, and molecular analysis of the CFTR gene. We then divided the patients into three groups by the number of mutations identified (none, one, and two or more) and compared those groups in terms of their characteristics. RESULTS: We evaluated 37 patients with phenotypic findings of CF, with or without sweat test confirmation. The mean age of the patients was 32.5 ± 13.6 years, and females predominated (75.7%). The molecular analysis contributed to the definitive diagnosis of CF in 3 patients (8.1%), all of whom had at least two mutations. There were 7 patients (18.9%) with only one mutation and 26 patients (70.3%) with no mutations. None of the clinical characteristics evaluated was found to be associated with the genetic diagnosis. The most common mutation was p.F508del, which was found in 5 patients. The combination of p.V232D and p.F508del was found in 2 patients. Other mutations identified were p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P, p.R1066H, and p.T351S. CONCLUSIONS: The molecular analysis of the CFTR gene coding region showed a limited contribution to the diagnostic investigation of patients suspected of having mild or atypical CF. In addition, there were no associations between the clinical characteristics and the genetic diagnosis.