Epigenetic regulation of transcription factor binding motifs promotes Th1 response in Chagas disease cardiomyopathy.

Chagas disease, caused by the protozoan Trypanosoma cruzi, is an endemic parasitic disease of Latin America, affecting 7 million people. Although most patients are asymptomatic, 30% develop complications, including the often-fatal Chronic Chagasic Cardiomyopathy (CCC). Although previous studies have demonstrated some genetic deregulations associated with CCCs, the causes of their deregulations remain poorly described. Based on bulk RNA-seq and whole genome DNA methylation data, we investigated the genetic and epigenetic deregulations present in the moderate and severe stages of CCC. Analysis of heart tissue gene expression profile allowed us to identify 1407 differentially expressed transcripts (DEGs) specific from CCC patients. A tissue DNA methylation analysis done on the same tissue has permitted the identification of 92 regulatory Differentially Methylated Regions (DMR) localized in the promoter of DEGs. An in-depth study of the transcription factors binding sites (TFBS) in the DMRs corroborated the importance of TFBS's DNA methylation for gene expression in CCC myocardium. TBX21, RUNX3 and EBF1 are the transcription factors whose binding motif appears to be affected by DNA methylation in the largest number of genes. By combining both transcriptomic and methylomic analysis on heart tissue, and methylomic analysis on blood, 4 biological processes affected by severe CCC have been identified, including immune response, ion transport, cardiac muscle processes and nervous system. An additional study on blood methylation of moderate CCC samples put forward the importance of ion transport and nervous system in the development of the disease.

Epigenetic regulation of transcription factor binding motifs promotes Th1 response in Chagas disease cardiomyopathy

Abstract: Chagas disease, caused by the protozoan Trypanosoma cruzi, is an endemic parasitic disease of Latin America, affecting 7 million people. Although most patients are asymptomatic, 30% develop complications, including the often-fatal Chronic Chagasic Cardiomyopathy (CCC). Although previous studies have demonstrated some genetic deregulations associated with CCCs, the causes of their deregulations remain poorly described. Based on bulk RNA-seq and whole genome DNA methylation data, we investigated the genetic and epigenetic deregulations present in the moderate and severe stages of CCC. Analysis of heart tissue gene expression profile allowed us to identify 1407 differentially expressed transcripts (DEGs) specific from CCC patients. A tissue DNA methylation analysis done on the same tissue has permitted the identification of 92 regulatory Differentially Methylated Regions (DMR) localized in the promoter of DEGs. An in-depth study of the transcription factors binding sites (TFBS) in the DMRs corroborated the importance of TFBS's DNA methylation for gene expression in CCC myocardium. TBX21, RUNX3 and EBF1 are the transcription factors whose binding motif appears to be affected by DNA methylation in the largest number of genes. By combining both transcriptomic and methylomic analysis on heart tissue, and methylomic analysis on blood, 4 biological processes affected by severe CCC have been identified, including immune response, ion transport, cardiac muscle processes and nervous system. An additional study on blood methylation of moderate CCC samples put forward the importance of ion transport and nervous system in the development of the disease.

Corrigendum: Polymorphisms in Genes Affecting Interferon-γ Production and Th1 T Cell Differentiation Are Associated With Progression to Chagas Disease Cardiomyopathy.

[This corrects the article DOI: 10.3389/fimmu.2020.01386.].

Polymorphisms in Genes Affecting Interferon-γ Production and Th1 T Cell Differentiation Are Associated With Progression to Chagas Disease Cardiomyopathy.

Background: Chagas disease, caused by the protozoan Trypanosoma cruzi, is endemic in Latin America. Thirty percent of infected individuals develop chronic Chagas cardiomyopathy (CCC), an inflammatory dilated cardiomyopathy that is the most important clinical consequence of T. cruzi infection, while the others remain asymptomatic (ASY). IFN-γ and IFN-γ-producing Th1-type T cells are increased in peripheral blood and CCC myocardium as compared to ASY patients, while the Th1-antagonizing cytokine IL-10 is more expressed in ASY patients. Importantly IFN-γ-producing Th1-type T cells are the most frequent cytokine-producing T cell subset in CCC myocardium, while expression of Th1-antagonizing cytokines IL-10 and IL-4 is unaltered. The control of IFN-γ production by Th1-type T cells may be a key event for progression toward CCC. A genetic component to disease progression was suggested by the familial aggregation of cases and the association of gene polymorphisms with CCC development. We here investigate the role of gene polymorphisms (SNPs) in several genes involved in the control of IFN-γ production and Th1 T cell differentiation in CCC development. Methods: We studied a Brazilian population including 315 CCC cases and 118 ASY subjects. We assessed 35 Tag SNPs designed to represent all the genetic information contained in the IL12B, IL10, IFNG, and IL4 genes. Results: We found 2 IL12 SNPs (rs2546893, rs919766) and a trend of association for a IL10 SNP (rs3024496) to be significantly associated with the ASY group. these associations were confirmed by multivariate analysis and allele tests. The rs919766C, 12rs2546893G, and rs3024496C alleles were associated to an increase risk to CCC development. Conclusions: Our data show that novel polymorphisms affecting IL12B and IL10, but not IFNG or IL4 genes play a role in genetic susceptibility to CCC development. This might indicate that the increased Th1 differentiation and IFN-γ production associated with CCC is genetically controlled.

Ansiedade, depressão e qualidade de vida de pacientes com câncer mamário e ginecológico sob quimioterapia / Ansiedad, depresión y calidad de vida de pacientes con cáncer mamario y ginecológico en quimioterapia / Anxiety, depression and quality of life of patients with breast or gynecological cancer undergoing chemotherapy1

RESUMO Tristeza e pesar são reações comuns entre pessoas que enfrentam o câncer, e estas reações podem influenciar sua qualidade de vida. Este estudo teve por objetivo verificar a existência de correlações entre os escores de ansiedade e depressão e domínios de qualidade de vida, nos momentos pré e pós-quimioterapia. Tratou-se de estudo observacional quantitativo, descritivo e de delineamento longitudinal-prospectivo. Participaram do estudo 14 mulheres, diagnosticadas com câncer de mama ou ginecológico, submetidas à quimioterapia, entre dezembro/2012 e abril/2013. Foram utilizados o WHOQOL-bref e Escala Hospitalar de Ansiedade e Depressão. A coleta de dados deu-se a partir da resposta individual das mulheres aos instrumentos. Para correlacionar os escores de ansiedade e depressão com os domínios de qualidade de vida, utilizou-se o coeficiente de correlação linear de Pearson. O domínio físico da qualidade de vida correlacionou-se negativamente com depressão, nos momentos pré e pós-quimioterapia. Houve também correlação negativa entre domínio psicológico e ambiental com ansiedade e depressão nos momentos pré e pós-quimioterapia. Em relação ao domínio social, não houve correlação. Concluiu-se que ansiedade e depressão influenciaram negativamente a qualidade de vida das mulheres estudadas nos domínios físico, psicológico e ambiental.

RESUMEN Tristeza y pesar son reacciones comunes entre personas que enfrentan el cáncer, y estas reacciones pueden afectar su calidad de vida. Este estudio tuvo el objetivo de verificar la existencia de correlaciones entre las puntuaciones de ansiedad y depresión y dominios de calidad de vida, en los momentos pre y post quimioterapia. Se trató de un estudio observacional cuantitativo, descriptivo y de delineamiento longitudinal-prospectivo. Participaron del estudio 14 mujeres, diagnosticadas con cáncer de mama o ginecológico, sometidas a quimioterapia, entre diciembre/2012 y abril/2013. Fueron utilizados el WHOQOL-bref y Escala Hospitalaria de Ansiedad y Depresión. La recolección de datos ocurrió a partir de la respuesta individual de las mujeres a los instrumentos. Para correlacionar las puntuaciones de ansiedad y depresión con los dominios de calidad de vida, se utilizó el coeficiente de correlación lineal de Pearson. El dominio físico de la calidad de vida se correlacionó negativamente con depresión, en los momentos pre y post quimioterapia. Hubo también correlación negativa entre dominio psicológico y ambiental con ansiedad y depresión en los momentos pre y post quimioterapia. Con relación al dominio social, no hubo correlación. Se concluyó que ansiedad y depresión influyen negativamente la calidad de vida de las mujeres estudiadas en los dominios físico, psicológico y ambiental.

ABSTRACT Sadness and grief are common reactions among people facing cancer, and these reactions can have an impact on their quality of life. The objective of this study was to determine the existence of correlations between depression and anxiety scores and quality of life domains, during pre- and post-chemotherapy. This was an observational, quantitative, descriptive study, with a longitudinal-prospective design. The 14 women who participated in the study had been diagnosed with breast or gynecological cancer and underwent chemotherapy between December 2012 and April 2013. The WHOQOL-BREF and Hospital Anxiety and Depression Scale were used. The data was collected through the individual responses of the women to the instruments. To correlate the anxiety and depression scores with the quality of life domains, the Pearson correlation coefficient was used. The physical health domain of quality of life had a negative correlation with depression during pre- and post-chemotherapy. There was also a negative correlation between the psychological and environment domains and anxiety and depression during pre- and post-chemotherapy. As for the social relationships domain, there was no correlation. It was concluded that anxiety and depression did not negatively influence the quality of life of the women in the physical health, psychological and environment domains.

Gene Polymorphisms in FAS (Rs3740286 and Rs4064) Are Involved in Endometriosis Development in Brazilian Women, but not those in CASP8 (rs13416436 and rs2037815) / Polimorfismos do gene FAS (rs3740286 e rs4064) estão envolvidos no desenvolvimento de endometriose em mulheres brasileiras, mas não os no CASP8 (rs13416436 e rs2037815)

Abstract Objective The present study aims to investigate the association between caspase-8 (CASP8) (rs13416436 and rs2037815) and Fas cell surface death receptor (FAS) (rs3740286 and rs4064) polymorphisms with endometriosis in Brazilian women. Methods In the present case-control study, 45 women with a diagnosis of endometriosis and 78 normal healthy women as a control group were included. The genotyping was determined by real-time polymerase chain reaction (PCR) with Taqman hydrolysis probes (Thermo Fisher Scientific, Darmstadt, Germany). Genotypic and allelic frequencies were analyzed using Chi-squared (χ2) test. In order to determine the inheritance models and haplotypes ,SNPStats (Institut Català d'Oncologia, Barcelona, Spain) was used. Levels of 5% (p = 0.05) were considered statistically significant. Results No significant difference was observed in genotypic or allelic frequencies between control and endometriosis groups for rs13416436 and rs2037815 (CASP8 gene). On the other hand, a significant difference between rs3740286 and rs4064 (FAS gene) was found. Regarding polymorphisms in the FAS gene, a statistically significant differencewas found in co-dominant and dominantmodels. Only the haplotype containing the rs3740286A and rs4064G alleles in the FAS gene were statistically significant. Conclusion The polymorphisms in the CASP8 gene were not associated with endometriosis. The results indicate an association between FAS gene polymorphisms and the risk of developing endometriosis.

Resumo Objetivo Investigar a associação entre os polimorfismos dos genes caspase-8 (CASP8) (rs13416436 e rs2037815) e FAS (rs3740286 e rs4064) em mulheres brasileiras com endometriose. Métodos Trata-se de um estudo do tipo caso-controle, no qual foram incluídas 45 mulheres com diagnóstico de endometriose e 78 controles. A genotipagem das amostras foi determinada usando a reação em cadeia de polimerase em tempo real com sondas de hidrólise TaqMan (Thermo Fisher Scientific, Darmstadt, Germany). As frequências genotípicas e alélicas foram analisadas usando o teste do qui-quadrado. O SNPStats (Institut Català d'Oncologia, Barcelona, Espanha) foi usado para determinar os modelos de herança e os haplótipos. Os níveis de significância estatística considerados foram de 5% (p = 0,05). Resultados Não foi observada diferença significativa nas frequências genotípicas ou alélicas entre os grupos de controle e de endometriose para os polimorfismos rs13416436 e rs2037815 (gene CASP8). Por outro lado, foi encontrada uma diferença significativa entre os polimorfismos rs3740286 e rs4064 (gene FAS). Em relação aos polimorfismos do gene FAS, foi encontrada uma diferença estatisticamente significativa nos modelos codominante e dominante. Apenas o haplótipo contendo os alelos rs3740286A e rs4064G no gene FAS foi estatisticamente significativo. Conclusão Não há associação entre os polimorfismos do gene CASP8 e endometriose. Entretanto, há associação entre os polimorfismos do gene FAS e o risco de desenvolver endometriose.

α-Tocopherol influences glycaemic control and miR-9-3 DNA methylation in overweight and obese women under an energy-restricted diet: a randomized, double-blind, exploratory, controlled clinical trial.

BACKGROUND: Excess weight is a strong risk factor for the development of dysglycaemia. It has been suggested that changes in the metabolism microRNAs, small non-coding RNAs that regulate gene expression, could precede late glycaemic changes. Vitamin E in turn may exert important functions in methylation and gene expression processes. This study aimed to determine the effect of α-tocopherol on glycaemic variables and miR-9-1 and miR-9-3 promoter DNA methylation in overweight women. METHODS: A randomized, double-blind, exploratory, placebo-controlled study was conducted in overweight and obese adult women (n = 44) who ingested synthetic vitamin E (all-rac-α-tocopherol), natural source vitamin E (RRR-rac-α-tocopherol) or placebo capsules and were followed up for a period of 8 weeks. Supplemented groups also received dietary guidance for an energy-restricted diet. An additional group that received no supplementation and did not follow an energy-restricted diet was also followed up. The intervention effect was evaluated by DNA methylation levels (quantitative real-time PCR assay) and anthropometric and biochemical variables (fasting plasma glucose, haemoglobin A1C, insulin, and vitamin E). RESULTS: Increased methylation levels of the miR-9-3 promoter region (P < 0.001) and reduced haemoglobin A1C (P < 0.05) were observed in the natural source vitamin E group after intervention. Increased fasting plasma glucose was observed in the synthetic vitamin E group, despite the significant reduction of anthropometric variables compared to the other groups. CONCLUSIONS: α-Tocopherol from natural sources increased methylation levels of the miR-9-3 promoter region and reduced haemoglobin A1C in overweight women following an energy-restricted diet. These results provide novel information about the influence of vitamin E on DNA methylation. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02922491. Registered 4 October, 2016.

Gene Polymorphisms in FAS (Rs3740286 and Rs4064) Are Involved in Endometriosis Development in Brazilian Women, but not those in CASP8 (rs13416436 and rs2037815).

OBJECTIVE: The present study aims to investigate the association between caspase-8 (CASP8) (rs13416436 and rs2037815) and Fas cell surface death receptor (FAS) (rs3740286 and rs4064) polymorphisms with endometriosis in Brazilian women. METHODS: In the present case-control study, 45 women with a diagnosis of endometriosis and 78 normal healthy women as a control group were included. The genotyping was determined by real-time polymerase chain reaction (PCR) with Taqman hydrolysis probes (Thermo Fisher Scientific, Darmstadt, Germany). Genotypic and allelic frequencies were analyzed using Chi-squared (χ2) test. In order to determine the inheritance models and haplotypes ,SNPStats (Institut Català d'Oncologia, Barcelona, Spain) was used. Levels of 5% (p = 0.05) were considered statistically significant. RESULTS: No significant difference was observed in genotypic or allelic frequencies between control and endometriosis groups for rs13416436 and rs2037815 (CASP8 gene). On the other hand, a significant difference between rs3740286 and rs4064 (FAS gene) was found. Regarding polymorphisms in the FAS gene, a statistically significant difference was found in co-dominant and dominant models. Only the haplotype containing the rs3740286A and rs4064G alleles in the FAS gene were statistically significant. CONCLUSION: The polymorphisms in the CASP8 gene were not associated with endometriosis. The results indicate an association between FAS gene polymorphisms and the risk of developing endometriosis.

OBJETIVO: Investigar a associação entre os polimorfismos dos genes caspase-8 (CASP8) (rs13416436 e rs2037815) e FAS (rs3740286 e rs4064) em mulheres brasileiras com endometriose. MéTODOS: Trata-se de um estudo do tipo caso-controle, no qual foram incluídas 45 mulheres com diagnóstico de endometriose e 78 controles. A genotipagem das amostras foi determinada usando a reação em cadeia de polimerase em tempo real com sondas de hidrólise TaqMan (Thermo Fisher Scientific, Darmstadt, Germany). As frequências genotípicas e alélicas foram analisadas usando o teste do qui-quadrado. O SNPStats (Institut Català d'Oncologia, Barcelona, Espanha) foi usado para determinar os modelos de herança e os haplótipos. Os níveis de significância estatística considerados foram de 5% (p = 0,05). RESULTADOS: Não foi observada diferença significativa nas frequências genotípicas ou alélicas entre os grupos de controle e de endometriose para os polimorfismos rs13416436 e rs2037815 (gene CASP8). Por outro lado, foi encontrada uma diferença significativa entre os polimorfismos rs3740286 e rs4064 (gene FAS). Em relação aos polimorfismos do gene FAS, foi encontrada uma diferença estatisticamente significativa nos modelos codominante e dominante. Apenas o haplótipo contendo os alelos rs3740286A e rs4064G no gene FAS foi estatisticamente significativo. CONCLUSãO: Não há associação entre os polimorfismos do gene CASP8 e endometriose. Entretanto, há associação entre os polimorfismos do gene FAS e o risco de desenvolver endometriose.

Perfil epidemiológico dos nascidos vivos em Belo Horizonte, MG, Brasil / Perfil epidemiológico de los nacidos vivos en Belo Horizonte, MG, Brasil / Epidemiological profile of live births in Belo Horizonte, MG, Brazil

OBJETIVO: este estudo teve por objetivo descrever o perfil epidemiológico dos nascidos vivos residentes no município de Belo Horizonte/MG, no período de 1994 a 2014. MÉTODO: trata-se de uma pesquisa transversal, retrospectiva, de abordagem quantitativa, realizada com base nos dados secundários disponíveis no Sistema de Informações sobre Nascidos Vivos. RESULTADOS: os resultados evidenciaram 738.314 nascimentos no município de Belo Horizonte/MG nesse período, sendo a maioria do sexo masculino (51,1%); a termo (90,2%); com índice de Apgar entre 08 e 10 no primeiro minuto de vida (85%); com peso ao nascer entre 2500g e 3999g (86,4%); e sem a presença de anomalias congênitas (58,2%). CONCLUSÃO: o Sistema de Informações sobre Nascidos Vivos representa um avanço no registro de dados e serve de subsídio para o planejamento e a implantação de políticas públicas voltadas para a saúde materno-infantil.

OBJECTIVE: this study aimed to describe the epidemiological profile of live births living in the city of Belo Horizonte, MG, from 1994 to 2014. METHOD: this is a cross-sectional, retrospective, quantitative approach based on the secondary data available in the Live Births Information System. RESULTS: the results showed 738,314 births in the city of Belo Horizonte / MG in this period, being the majority male ones (51.1%); at term (90.2%); with an Apgar score between 8 and 10 in the first minute of life (85%); with birth weight between 2500g and 3999g (86.4%); and without the presence of congenital anomalies (58.2%). CONCLUSION: the Live Births Information System represents an advance in the data registry and works as a subsidy for the planning and implementation of public policies focused on maternal and child health.

OBJETIVO: este estudio tuvo por objetivo describir el perfil epidemiológico de los nacidos vivos residentes en el municipio de Belo Horizonte/MG, en el período de 1994 a 2014. MÉTODO: se trata de una pesquisa transversal, retrospectiva, de abordaje cuantitativa, realizada con base en los datos secundarios disponibles en el Sistema de Informaciones sobre Nacidos Vivos. RESULTADOS: los resultados evidenciaron 738.314 nacimientos en el municipio de Belo Horizonte/MG en ese período, siendo la mayoría del sexo masculino (51,1%); a término (90,2%); con índice de Apgar entre 08 y 10 en el primero minuto de vida (85%); con peso al nacer entre 2500g y 3999g (86,4%); y sin la presencia de anomalías congénitas (58,2%). CONCLUSIÓN: el Sistema de Informaciones sobre Nacidos Vivos representa un avanzo en el registro de datos y sirve de subsidio para el planeamiento y la implantación de políticas públicas volteadas para la salud materno-infantil.

CASPASE-8 gene polymorphisms (rs13416436 and rs2037815) are not associated with preeclampsia development in Brazilian women.

BACKGROUND: Preeclampsia is responsible for considerable mortality and morbidity of mother and sibling. The etiology of preeclampsia is still unknown. Family studies indicate the involvement of genes located on chromosome 2 in preeclampsia development. Considering the importance of apoptosis and chromosome 2, one promising candidate for the study of the genetic cause of this syndrome is the CASPASE-8 gene, which was chosen as the subject of this study. OBJECTIVE: The aim of this study was to determine the frequencies of the genotypes for CASP8 gene polymorphisms (rs13416436 and rs2037815) and to associate these with preeclampsia development in Brazilian women. METHODS: Women with and without preeclampsia were investigated. Accordingly, peripheral blood was collected and DNA extracted, followed by genotyping using Real-time PCR with hydrolysis probe (Taqman® Life Technologies). RESULTS: The results showed no association between genotypes and preeclampsia development for both polymorphisms studied (χ2 = 1.03; p = 0.59, for rs13416436 and χ2 = 1.06; p = 0.58 for rs2037815). CONCLUSIONS: It seems that CASP8 gene polymorphisms (rs13416436 and rs2037815) are not important candidates for the development of preeclampsia. Other genes related to the apoptosis process or other polymorphisms in this gene should be studied in order to understand better the etiology of preeclampsia.

Polymorphism in the lymphotoxin-alpha gene, position +252 (rs909253), is not associated with preeclampsia development in Brazilian women.

PURPOSE: To investigate the frequencies of polymorphic allele and genotypes for the LT-α gene, position +252 (rs909253), in Brazilian women with preeclampsia. METHODS: This is a case-control study, in which 30 women with preeclampsia, classified according to the criteria of the National High Blood Pressure Education Program, and 115 women in the control group, with at least two healthy pregnancies, were selected. Peripheral blood was collected, and DNA was extracted, followed by genotyping, using specific primers and restriction analysis. The genotypes obtained were AA, AG and GG. Statistical analysis was performed using the χ2 association test. The Hardy-Weinberg Equilibrium was tested using the Haploview Program. RESULTS: The results showed no association between genotypes and preeclampsia development (χ2=2.0; p=0.4). When the AG and GG genotypes were grouped according to allele G presence or absence (genotype AA), the data showed that the presence of allele G was not significantly different between cases (women with preeclampsia) and controls (χ2=0.0; p=1.0). The LT-α gene polymorphism, position +252 (rs909253), seems not to be an important candidate for the development of preeclampsia. Other inflammatory genes should be researched, and studies involving gene-environment interactions should be performed, in order to reach a better understanding of the etiology of the preeclampsia.

Polymorphism in the lymphotoxin-alpha gene, position +252 (rs909253), is not associated with preeclampsia development in Brazilian women / Polimorfismo no gene da linfotoxina alfa, posição +252 (rs909253), não está associado com o desenvolvimento de pré-eclâmpsia em mulheres brasileiras

PURPOSE: To investigate the frequencies of polymorphic allele and genotypes for the LT-α gene, position +252 (rs909253), in Brazilian women with preeclampsia.METHODS: This is a case-control study, in which 30 women with preeclampsia, classified according to the criteria of the National High Blood Pressure Education Program, and 115 women in the control group, with at least two healthy pregnancies, were selected. Peripheral blood was collected, and DNA was extracted, followed by genotyping, using specific primers and restriction analysis. The genotypes obtained were AA, AG and GG. Statistical analysis was performed using the χ2association test. The Hardy-Weinberg Equilibrium was tested using the Haploview Program.RESULTS: The results showed no association between genotypes and preeclampsia development (χ2=2.0; p=0.4). When the AG and GG genotypes were grouped according to allele G presence or absence (genotype AA), the data showed that the presence of allele G was not significantly different between cases (women with preeclampsia) and controls (χ2=0.0; p=1.0). The LT-α gene polymorphism, position +252 (rs909253), seems not to be an important candidate for the development of preeclampsia. Other inflammatory genes should be researched, and studies involving gene-environment interactions should be performed, in order to reach a better understanding of the etiology of the preeclampsia.

OBJETIVO: Investigar as frequências do alelo polimórfico e genótipos para o gene da LT-α, posição +252 (rs909253), em mulheres brasileiras com pré-eclâmpsia.MÉTODOS: Trata-se de um estudo caso-controle, em que 30 mulheres com pré-eclâmpsia, classificadas de acordo com os critérios do National High Blood Pressure Education Program, e 115 mulheres do grupo controle, com pelo menos duas gestações saudáveis, foram selecionadas. Amostra de sangue periférico foi colhida, e o DNA foi extraído, seguido pela genotipagem, usando iniciadores específicos e análise de restrição. Os genótipos obtidos foram AA, AG e GG. A análise estatística foi realizada utilizando-se o teste de associação χ2. O Equilíbrio de Hardy-Weinberg foi testado com o auxílio do programa Haploview.RESULTADOS: Os resultados não mostraram associação entre os genótipos e o desenvolvimento de pré-eclâmpsia (χ2=2,0; p=0,4). Quando os genótipos AG e GG foram agrupados de acordo com a presença do alelo G ou ausência (genótipo AA), os dados mostraram que a presença do alelo G não foi significativamente diferente entre casos (mulheres com pré-eclâmpsia) e controles (χ2=0,0; p=1,0). O polimorfismo no gene LT-α, posição +252 (rs909253), parece não ser um importante candidato para o desenvolvimento de pré-eclâmpsia. Outros genes inflamatórios devem ser pesquisados, e estudos envolvendo interações gene-ambiente devem ser realizados para que se possa alcançar um melhor entendimento da etiologia da pré-eclâmpsia.

Functional IL18 polymorphism and susceptibility to Chronic Chagas Disease.

BACKGROUND: Chronic Chagas Disease cardiomyopathy (CCC), a life-threatening inflammatory dilated cardiomyopathy, affects 30% of the approximately 8 million patients infected by Trypanosoma cruzi, the rest of the infected subjects remaining asymptomatic (ASY). The Th1 T cell-rich myocarditis plays a pivotal role in CCC pathogenesis. Local expression of IL-18 in CCC myocardial tissue has recently been described. IL-18 could potentially amplify the process by inducing increased expression of IFN-γ which in turn can increase the production of IL-18, thereby creating a positive feedback mechanism. In order to assess the contribution of the IL-18 to susceptibility to Chronic Chagas Disease, we investigated the association between a single nucleotide polymorphism (SNP) located in the IL-18 gene with the risk of developing Chagas cardiomyopathy. METHODS AND RESULTS: We analyzed the rs2043055 marker in the IL18 gene in a cohort of Chagas disease cardiomyopathy patients (n=849) and asymptomatic subjects (n=202). We found a significant difference in genotype frequencies among moderate and severe CCC patients with ventricular dysfunction. CONCLUSIONS: Our analysis suggests that the IL18 rs2043055 polymorphism- or a SNP in tight linkage disequilibrium with it- may contribute to modulating the Chagas cardiomyopathy outcome.

Functional IL18 polymorphism and susceptibility to Chronic ChagasDisease

Background: Chronic Chagas Disease cardiomyopathy (CCC), a life-threatening inflammatory dilated cardiomyopathy,affects 30% of the approximately 8 million patients infected by Trypanosoma cruzi, the restof the infected subjects remaining asymptomatic (ASY). The Th1 T cell-rich myocarditis plays a pivotalrole in CCC pathogenesis. Local expression of IL-18 in CCC myocardial tissue has recently been described.IL-18 could potentially amplify the process by inducing increased expression of IFN-c which in turn canincrease the production of IL-18, thereby creating a positive feedback mechanism. In order to assess thecontribution of the IL-18 to susceptibility to Chronic Chagas Disease, we investigated the associationbetween a single nucleotide polymorphism (SNP) located in the IL-18 gene with the risk of developingChagas cardiomyopathy.Methods and results: We analyzed the rs2043055 marker in the IL18 gene in a cohort of Chagas diseasecardiomyopathy patients (n = 849) and asymptomatic subjects (n = 202). We found a significant differencein genotype frequencies among moderate and severe CCC patients with ventricular dysfunction.Conclusions: Our analysis suggests that the IL18 rs2043055 polymorphism- or a SNP in tight linkagedisequilibrium with it- may contribute to modulating the Chagas cardiomyopathy outcome.

[Protective role of the G allele of the polymorphism in the Interleukin 10 gene (-1082G/A) against the development of preeclampsia]. / Papel protetor do alelo G do polimorfismo no gene Interleucina 10 (-1082G/A) contra o desenvolvimento de pré-eclâmpsia.

PURPOSE: To identify the frequency of polymorphism in the IL-10 gene, rs1800896 (-1082 A/G), in women with preeclampsia (PE) and in women in a control group and to associate the presence of this polymorphism with protection against the development of PE. METHODS: This was a case-control study conducted on 54 women with PE, classified according to the criteria of the National High Blood Pressure Education Program, and on 172 control women with at least two healthy pregnancies. The proposed polymorphism was studied by the technique of real time polymerase chain reaction (qPCR), with hydrolysis probes. Statistical analysis was performed using the χ2 test. Odds ratio and confidence interval of 95% were used to measure the strength of association between the studied polymorphism and the development of PE. RESULTS: Statistically increased frequency of the AG genotype was observed among control women (85 versus 15% in women with PE). The G allele was significantly more frequent among control women than PE women (χ2 test, p = 0.01). The odds ratio for carriers of the G allele was 2.13, indicating a lower risk of developing PE compared to non-carriers. CONCLUSIONS: Thus, an association is suggested to occur between the presence of the G allele of the polymorphism in the IL-10 rs1800896 (-1082 A/G) gene and protection against the development of PE. More studies investigating the contribution of these variations and the mechanisms by which they affect the risk of developing PE still need to be undertaken.

Papel protetor do alelo G do polimorfismo no gene Interleucina 10 (-1082G/A) contra o desenvolvimento de pré-eclâmpsia / Protective role of the G allele of the polymorphism in the Interleukin 10 gene (-1082G/A) against the development of preeclampsia

OBJETIVOS: Identificar a frequência do polimorfismo no gene IL-10, rs1800896 (-1082 A/G), em mulheres com pré-eclâmpsia (PE) e em mulheres do grupo controle e associar a presença deste polimorfismo com a proteção contra o desenvolvimento da PE. MÉTODOS: Estudo do tipo caso-controle, no qual foram selecionadas 54 mulheres com PE, classificadas de acordo com os critérios da National High Blood Pressure Education Program e 172 mulheres do grupo controle, com pelo menos duas gestações saudáveis. O polimorfismo proposto foi estudado utilizando-se a técnica de reação em cadeia da polimerase em tempo real (qPCR), com sondas de hidrólise. A análise estatística foi realizada utilizando-se o teste de associação do χ2. Odds ratio e seu intervalo de confiança de 95% foram usados para medir a força de associação entre o polimorfismo estudado e o desenvolvimento da PE. RESULTADOS: Foi observado aumento significativo da frequência do genótipo AG entre mulheres do grupo controle (85 versus 15% nas mulheres com PE). O alelo G é significativamente mais frequente entre as mulheres do grupo controle do que nas com PE (Teste χ2; p=0,01). O odds ratio para as portadoras do alelo G foi de 2, 13, indicando que apresentam menor risco de desenvolver PE do que as não portadoras. CONCLUSÕES: Sugere-se associação entre a presença do alelo G do polimorfismo no gene IL-10, rs1800896 (-1082 A/G), e a proteção contra o desenvolvimento da PE. Mais estudos sobre a contribuição dessas variações e os mecanismos pelos quais afetam o risco de desenvolver PE ainda necessitam de ser realizadas. .

PURPOSE: To identify the frequency of polymorphism in the IL-10 gene, rs1800896 (-1082 A/G), in women with preeclampsia (PE) and in women in a control group and to associate the presence of this polymorphism with protection against the development of PE. METHODS: This was a case-control study conducted on 54 women with PE, classified according to the criteria of the National High Blood Pressure Education Program, and on 172 control women with at least two healthy pregnancies. The proposed polymorphism was studied by the technique of real time polymerase chain reaction (qPCR), with hydrolysis probes. Statistical analysis was performed using the χ2 test. Odds ratio and confidence interval of 95% were used to measure the strength of association between the studied polymorphism and the development of PE. RESULTS: Statistically increased frequency of the AG genotype was observed among control women (85 versus 15% in women with PE). The G allele was significantly more frequent among control women than PE women (χ2 test, p = 0.01). The odds ratio for carriers of the G allele was 2.13, indicating a lower risk of developing PE compared to non-carriers. CONCLUSIONS: Thus, an association is suggested to occur between the presence of the G allele of the polymorphism in the IL-10 rs1800896 (-1082 A/G) gene and protection against the development of PE. More studies investigating the contribution of these variations and the mechanisms by which they affect the risk of developing PE still need to be undertaken. .

Polimorfismos no gene VEGF para o desenvolvimento das síndromes hipertensivas gestacionais: revisão de literature / Polimorfismos en el gen VEGF para el desarrollo de los síndromes hipertensivos de la gestación: una revisión de la literatura / Polymorphisms in the VEGF gene to the development of gestational hypertensive syndromes: a lite-rature review

Estudos de genes de suscetibilidade para SíndromesHipertensivas Gestacionais (SGH) sãobaseados nas hipóteses mais aceitas sobre sua etiopatogênese, como a isquemia placentária e adisfunção endotelial. Na gestação, o Fator de Crescimento do Endotélio Vascular (VEGF)regula a angiogênese e atua no remodelamento da artéria espiralada. Esta revisão bibliográficaavaliou a associação de polimorfismos genéticos nogeneVEGFcom as SHG, no período de2004 a 2013, utilizando o Pubmed como fonte de pesquisa. Os polimorfismos +405G/C e -2578C/A foram associados à redução do risco de desenvolver pré-eclâmpsia (PE), +936C/Tconfere risco maior para desenvolvimento e gravidade de PE. Os polimorfismos rs7664413,rs1485766 e rs6838834 estão associados ao risco dedesenvolvimento de PE apenas emmulheres brancas e mulheres negras, respectivamente. Assim, alguns resultados mostraram-secontraditórios, devido ao fato de que estudos envolvendo polimorfismos genéticos sãocomplexos e podem ser válidos em uma população, masnão em outra .

Studies of susceptibility genes for Gestational Hypertensive Syndromes (SGH) are based onthe most accepted hypotheses about its etiopathogenesis, such as placental ischemia andendothelial dysfunction. During pregnancy, the Factor Vascular Endothelial Growth (VEGF)regulates angiogenesis and acts in spiral artery remodeling. This literature review examinedthe association of genetic polymorphisms in theVEGFgene with the SHG in the period 2004-2013, using Pubmed as a research resource. Polymorphisms +405 G/C and-2578 C/A wereassociated with reduced risk of developing preeclampsia (PE), +936 C/T confers increasedrisk for the development and severity of PE. The polymorphisms rs7664413, rs1485766 and rs6838834 are associated with the risk of developing PE only in white women and blackwomen, respectively. Thus, some contradictory results were due to the fact that studiesinvolving genetic polymorphisms are complex and maybe valid in a population but not inanother .

Estudios de genes de susceptibilidad para SíndromesHipertensivas Gestacionales (SGH) sonbasados en las hipótesis más aceptas sobre su etiopatogenia, como la isquemia placentaria y ladisfunción endotelial. En la gestación, el Factor de Crecimiento del Endotelio Vascular(VEGF) regula la angiogenese y actúa en el remodelamiento de la arteria espiralada. Estarevisión bibliográfica evaluó la asociación de polimorfismos genéticos en el geneVEGFconlas SHG, en el período de 2004 hasta 2013, utilizando el Pubmed como fuente de pesquisa.Los polimorfismos +405G/C e -2578C/A fueron asociados a la reducción del riesgo dedesarrollar pre-eclámpsia (PE), +936C/T confiere riesgo mayor para desenvolvimiento ygravedad de PE. Los polimorfismos rs7664413, rs1485766 y rs6838834 están asociados alriesgo de desenvolvimiento de PE apenas en mujeresblancas y mujeres negras,respectivamente. Así, algunos resultados se muestrearon contradictorios, debido al facto deque estudios envolviendo polimorfismos genéticos son complejos y pueden ser válidos en unapoblación, pero no en otra .

Genetic susceptibility to Chagas disease cardiomyopathy: involvement of several genes of the innate immunity and chemokine-dependent migration pathways.

BACKGROUND: Chagas disease, caused by the protozoan Trypanosoma cruzi is endemic in Latin America. Thirty percent of infected individuals develop chronic Chagas cardiomyopathy (CCC), an inflammatory dilated cardiomyopathy that is, by far, the most important clinical consequence of T. cruzi infection. The others remain asymptomatic (ASY). A possible genetic component to disease progression was suggested by familial aggregation of cases and the association of markers of innate and adaptive immunity genes with CCC development. Migration of Th1-type T cells play a major role in myocardial damage. METHODS: Our genetic analysis focused on CCR5, CCL2 and MAL/TIRAP genes. We used the Tag SNPs based approach, defined to catch all the genetic information from each gene. The study was conducted on a large Brazilian population including 315 CCC cases and 118 ASY subjects. RESULTS: The CCL2rs2530797A/A and TIRAPrs8177376A/A were associated to an increase susceptibility whereas the CCR5rs3176763C/C genotype is associated to protection to CCC. These associations were confirmed when we restricted the analysis to severe CCC, characterized by a left ventricular ejection fraction under 40%. CONCLUSIONS: Our data show that polymorphisms affecting key molecules involved in several immune parameters (innate immunity signal transduction and T cell/monocyte migration) play a role in genetic susceptibility to CCC development. This also points out to the multigenic character of CCC, each polymorphism imparting a small contribution. The identification of genetic markers for CCC will provide information for pathogenesis as well as therapeutic targets.

Polymorphism in the alpha cardiac muscle actin 1 gene is associated to susceptibility to chronic inflammatory cardiomyopathy.

AIMS: Chagas disease, caused by the protozoan Trypanosoma cruzi is endemic in Latin America, and may lead to a life-threatening inflammatory dilated, chronic Chagas cardiomyopathy (CCC). One third of T. cruzi-infected individuals progress to CCC while the others remain asymptomatic (ASY). A possible genetic component to disease progression was suggested by familial aggregation of cases and the association of markers of innate and adaptive immunity genes with CCC development. Since mutations in multiple sarcomeric genes, including alpha-cardiac actin (ACTC1) have been involved in hereditary dilated cardiomyopathy, we investigated the involvement of the ACTC1 gene in CCC pathogenesis. METHODS AND RESULTS: We conducted a proteomic and genetic study on a Brazilian study population. The genetic study was done on a main cohort including 118 seropositive asymptomatic subjects and 315 cases and the replication was done on 36 asymptomatic and 102 CCC cases. ACTC1 protein and mRNA levels were lower in myocardial tissue from patients with end-stage CCC than those found in hearts from organ donors. Genotyping a case-control cohort of CCC and ASY subjects for all informative single nucleotide polymorphism (SNP) in the ACTC1 gene identified rs640249 SNP, located at the 5' region, as associated to CCC. Associations are borderline after correction for multiple testing. Correlation and haplotype analysis led to the identification of a susceptibility haplotype. Functional assays have shown that the rs640249A/C polymorphism affects the binding of transcriptional factors in the promoter regions of the ACTC1 gene. Confirmation of the detected association on a larger independent replication cohort will be useful. CONCLUSIONS: Genetic variations at the ACTC1 gene may contribute to progression to chronic Chagas Cardiomyopathy among T. cruzi-infected patients, possibly by modulating transcription factor binding to ACTC1 promoter regions.

Association between the lymphotoxin-alpha gene polymorphism and chagasic cardiopathy.

Lymphotoxin-alpha (LT-alpha or LTA) is an inflammatory cytokine that is involved in the organization and maintenance of the inflammatory process and in the arrangement of cells at the site of inflammation. These features suggest an important role in the development of chronic Chagas' disease, especially the cardiac form. The objective of this study was to evaluate LT-alpha genetics and its biological role in chronic Chagas' disease. A total of 284 subjects were studied. The LT-alpha single-nucleotide polymorphism (+252) was analyzed by the polymerase chain reaction-restriction fragment length polymorphism and expression by enzyme-linked immunosorbent assay in culture supernatants and in individual T lymphocytes by flow cytometry. The risk of developing the cardiac form was 2.8 times higher among carriers of genotype GG and 2.4 times among carriers of genotype GA when compared to subjects carrying genotype AA. Seropositive subjects carrying the G allele produced significantly higher levels of LT-alpha. The cytokine was mainly expressed by CD8(+) T lymphocytes in the absence of any stimulus and after stimulation with the Trypanosoma cruzi antigen. This study provides genetic and biological evidence for an important role of LT-alpha in the development of the cardiac form of Chagas' disease.