Tumor necrosis factor-α and interleukin-6 expression in leukocytes and their association with polymorphisms and bone markers in diabetic individuals treated with pioglitazone.

BACKGROUND: Pioglitazone is a peroxisome proliferator-activated receptor gamma (PPARγ) activator used in the treatment of type 2 diabetes (DM2) patients and it has been suggested that can induce bone loss. Tumor necrosis factor-α (TNFα) and interleukin-6 (IL-6) mRNA expression in blood leukocytes and the relationship with polymorphisms and bone markers in DM2 treated with pioglitazone were investigated. METHODS: DM2 (n=53) and normoglycemic (NG, n=52) individuals were included. DM2 patients were treated with pioglitazone (45 mg/day/16 weeks). mRNA expression was evaluated by real-time polymerase chain reaction (PCR). TNFA -308G>A and IL6 -174G>C polymorphisms were detected by PCR-RFLP and high resolution melting polymerase chain reaction (HRM-PCR). RESULTS: Pioglitazone reduced bone specific alkaline phosphatase (bALP) and increased TNFα in DM2 group (p<0.001). DM2 or pioglitazone did not influence TNFα and IL-6 expression (p>0.05). TNFA -308A allele was associated with reduced basal TNFα mRNA levels in NG and DM2 and reduced alkaline phosphatase (tALP) after treatment (p<0.05). IL6 -174C allele was associated with decreased oral glucose tolerance test (OGTT)-2 h in DM2 individuals (p<0.05). CONCLUSIONS: TNFA -308G >A polymorphism appear to be involved in regulation of gene expression independently of hyperglycemia and its interaction with pioglitazone may modify tALP, a important bone marker. IL6 -174G>C variant is related with reduced risk of postprandial hyperglycemia but not with mRNA expression or bone markers.

High serum miR-421 is associated with metabolic dysregulation and inflammation in patients with metabolic syndrome.

Aim: To explore the association of circulating miRNAs with adiposity, metabolic status and inflammatory biomarkers in patients with metabolic syndrome (MetS). Methods: Serum levels of 372 miRNAs were measured in patients with (n = 6) and without MetS (n = 6) by quantitative PCR array, and dysregulated miRNAs were validated in a larger cohort (MetS, n = 89; non-MetS, n = 144). Results: In the screening study, seven miRNAs were dysregulated in patients with MetS, and miR-421 remained increased in the validation study. miR-421 was associated with a high risk of MetS and insulin resistance and hypertension and correlated with glycated hemoglobin, triacylglycerols, high-sensitivity CRP, IL-6, resistin and adiponectin (p < 0.05). Conclusion: Circulating miR-421 is a potential biomarker for insulin resistance, metabolic dysregulation and inflammatory status in patients with MetS.

Novel genes detected by transcriptional profiling from whole-blood cells in patients with early onset of acute coronary syndrome.

BACKGROUND: Genome-wide expression analysis using microarrays has been used as a research strategy to discovery new biomarkers and candidate genes for a number of diseases. We aim to find new biomarkers for the prediction of acute coronary syndrome (ACS) with a differentially expressed mRNA profiling approach using whole genomic expression analysis in a peripheral blood cell model from patients with early ACS. METHODS AND RESULTS: This study was carried out in two phases. On phase 1 a restricted clinical criteria (ACS-Ph1, n=9 and CG-Ph1, n=6) was used in order to select potential mRNA biomarkers candidates. A subsequent phase 2 study was performed using selected phase 1 markers analyzed by RT-qPCR using a larger and independent casuistic (ACS-Ph2, n=74 and CG-Ph2, n=41). A total of 549 genes were found to be differentially expressed in the first 48 h after the ACS-Ph1. Technical and biological validation further confirmed that ALOX15, AREG, BCL2A1, BCL2L1, CA1, COX7B, ECHDC3, IL18R1, IRS2, KCNE1, MMP9, MYL4 and TREML4, are differentially expressed in both phases of this study. CONCLUSIONS: Transcriptomic analysis by microarray technology demonstrated differential expression during a 48 h time course suggesting a potential use of some of these genes as biomarkers for very early stages of ACS, as well as for monitoring early cardiac ischemic recovery.

Relationship of short tandem repeats flanking leptin-melanocortin pathway genes with anthropometric profile and leptinemia in Brazilian individuals.

OBJECTIVE: To investigate the relationship of short tandem repeats (STR) near genes involved in the leptin-melanocortin pathway with body mass index (BMI) and leptinemia. SUBJECTS AND METHODS: Anthropometric variables and leptinemia were measured in 100 obese and 110 nonobese individuals. D1S200, D2S1788, DS11912, and D18S858 loci were analyzed by PCR and high-resolution electrophoresis. RESULTS: Overall STR allele frequencies were similar between the obese and non-obese group (p > 0.05). Individual alleles D1S200 (17), D11S912 (43), D18S858 (11/12) were associated with obesity (p < 0.05). Individuals carrying these alleles showed higher BMI than non-carriers (p < 0.05). Moreover, a relationship between D18S858 11/12 alleles and increased waist circumference was found (p = 0.040). On the other hand, leptinemia was not influenced by the studied STRs (p > 0.05). CONCLUSIONS: D1S200, D11S912, and D18S858 loci are associated with increased BMI and risk for obesity in this sample.

Relationship between variants of the leptin gene and obesity and metabolic biomarkers in Brazilian individuals.

OBJECTIVE: The relationship between variants of the leptin gene (LEP) and obesity and metabolic biomarkers was investigated in Brazilian individuals. SUBJECTS AND METHODS: One-hundred-ten obese (BMI > 30 kg/m(2)) and 100 non-obese individuals (145 women and 65 men, aged 49 +/- 14 years) were randomly selected. Plasma leptin, glycemia, serum lipid measurements and LEP -2548G>A and 3'HVR polymorphisms were analyzed. RESULTS: The LEP -2548GG genotype was associated with a 2.2% and 2.0% increase in BMI (p = 0.009) and plasma leptin (p = 0.031), respectively. 3'HVR I/II (classes I/I+I/II) genotypes contributed with 1.8% of BMI values (p = 0.046). LEP I/G combined genotypes (I/IGG, I/IGA and I/IIGG) were associated with obesity, and increased BMI, waist circumference, leptin and triglycerides (p < 0.05). These relationships were found in women (p < 0.05) but not in men. LEP I/G combined genotypes were not associated with hypertension, hyperglycemia, dyslipidemia and coronary artery disease. CONCLUSIONS: LEP I/G combined genotypes are associated with obesity-related metabolic biomarkers and phenotype in a gender-dependent manner.

Detection of the TLR4 1196C>T polymorphism by mismatched-polymerase chain reaction using plasmid DNA as internal control in restriction fragment length polymorphism assays.

BACKGROUND: Restriction fragment length polymorphism (RFLP) is a common molecular assay used for genotyping, and it requires validated quality control procedures to prevent mistyping caused by impaired endonuclease activity. We have evaluated the usefulness of a plasmid-based internal control in RFLP assays. RESULTS: Blood samples were collected from 102 individuals with acute myocardial infarction (AMI) and 108 non-AMI individuals (controls) for DNA extraction and laboratory analyses. The 1196C > T polymorphism in the toll-like receptor 4 (TLR4) gene was amplified by mismatched-polymerase chain reaction (PCR). Amplicons and pBluescript II SK- plasmid were simultaneously digested with endonuclease HincII. Fragments were separated on 2% agarose gels. Plasmid was completely digested using up to 55.2 nmL/L DNA solutions and 1 microL PCR product. Nevertheless, plasmid DNA with 41.4 nM or higher concentrations was incompletely digested in the presence of 7 microL PCR product. In standardized conditions, TLR4 1196C>T variant was accurately genotyped. TLR4 1196T allele frequency was similar between AMI (3.1%) and controls (2.0%, p = 0.948). TLR4 SNP was not associated with AMI in this sample population. In conclusion, the plasmid-based control is a useful approach to prevent mistyping in RFLP assays, and it is validate for genetic association studies such as TLR4 1196C>T.

Leptin G-2548A promoter polymorphism is associated with increased plasma leptin and BMI in Brazilian women.

Variants in leptin gene (LEP) have been implicated in the pathogenesis of obesity. The relationship between LEP G-2548A polymorphism and obesity-related traits was evaluated in a sample of Brazilian women (n = 228) who were randomly selected from two clinical centers in Sao Paulo city. Blood samples were collected for DNA extraction, plasma leptin and serum lipids measurements. LEP G-2548A genotypes were identified by a PCR- RFLP strategy using the endonuclease Alw44I. LEP G-2548A was associated with obesity after adjustment for covariates (age, hypertension, coronary artery disease, smoking and physical activity). Women carrying G allele had a four times higher risk of obesity than the A allele carriers (OR: 4.11, CI95%: 1.06-15.90, p = 0.041). G allele was also related to increased plasma leptin (p = 0.024) and body mass index (p = 0.027). Hypertension, hyperglycemia, dyslipidemia and coronary artery disease were associated with obesity. However LEP G-2548A polymorphism was not related to these variables. All together these data suggest that LEP G-2548A polymorphism has an important role in regulating plasma leptin levels and body mass index in women.

Relationship of short tandem repeats flanking leptin-melanocortin pathway genes with anthropometric profile and leptinemia in Brazilian individuals / Relação de short tandem repeats (STR) em genes envolvidos na via da leptina-melanocortina com o perfil antropométrico e a leptinemia em brasileiros

OBJECTIVE: To investigate the relationship of short tandem repeats (STR) near genes involved in the leptin-melanocortin pathway with body mass index (BMI) and leptinemia. SUBJECTS AND METHODS: Anthropometric variables and leptinemia were measured in 100 obese and 110 nonobese individuals. D1S200, D2S1788, DS11912, and D18S858 loci were analyzed by PCR and high-resolution electrophoresis. RESULTS: Overall STR allele frequencies were similar between the obese and non-obese group (p > 0.05). Individual alleles D1S200 (17), D11S912 (43), D18S858 (11/12) were associated with obesity (p < 0.05). Individuals carrying these alleles showed higher BMI than non-carriers (p < 0.05). Moreover, a relationship between D18S858 11/12 alleles and increased waist circumference was found (p = 0.040). On the other hand, leptinemia was not influenced by the studied STRs (p > 0.05). CONCLUSIONS: D1S200, D11S912, and D18S858 loci are associated with increased BMI and risk for obesity in this sample.

OBJETIVO: Investigar a relação de short tandem repeats (STR) em genes envolvidos na via da leptina-melanocortina com índice de massa corporal (IMC) e leptinemia. SUJEITOS E MÉTODOS: Variáveis antropométricas e leptinemia foram medidas em 100 indivíduos obesos e 110 não obesos. Os loci D1S200, D2S1788, DS11912 e D18S858 foram analisados por PCR e eletroforese de alta resolução. RESULTADOS: As frequências globais dos alelos da STR foram similares entre os grupos obeso e não obeso (p > 0,05). Alelos individuais de D1S200 (17), D11S912 (43), D18S858 (11/12) foram associados com obesidade (p < 0,05). Indivíduos portadores desses alelos apresentaram valores de IMC maiores que os dos não portadores (p < 0,05). Além disso, a presença dos alelos D18S858 11/12 foi relacionada com circunferência abdominal elevada (p = 0,040). Por outro lado, a leptinemia não foi influenciada pelos STRs estudados (p > 0,05). CONCLUSÕES: Os loci D1S200, D11S912 e D18S858 são associados com IMC aumentado e risco de obesidade nesta amostra populacional.

LEP 3'HVR is associated with obesity and leptin levels in Brazilian individuals.

Leptin plays an important role in satiety signaling and is related to obesity. Variants of leptin gene (LEP) have been associated to differences in plasma leptin levels and obesity-related phenotypes. The purpose of this study was to evaluate the association of LEP 3'HVR and leptin concentrations and obesity-related traits in our population. Anthropometrics and systolic/diastolic pressure were measured in 210 unrelated Brazilian individuals. Blood samples were collected for quantification of leptin, glucose and lipids and DNA extraction. LEP 3'HVR polymorphic region was amplified by PCR and fragments were analyzed by polyacrylamide gel electrophoresis. Obesity was associated with hypertension, hyperglycemia, obesity-related traits, plasma leptin and serum lipids (p < 0.05). The frequency of LEP 3'HVR class I alleles (I/I + I/II genotypes) was higher in obese (p = 0.043) than in non-obese individuals. Multivariate logistic regression showed that the risk for obesity is nine times higher in hypertensive individuals and two times higher in class I alleles carriers. The presence of class I alleles was associated with increased BMI and WC. Plasma leptin was related to class I alleles in women (p < 0.05). No association was found between LEP 3'HVR and hypertension or risk factors for CAD in our sample. Our results suggest that LEP 3'HVR is an important predictor for obesity-related traits and leptin plasma levels.

Relationship between variants of the leptin gene and obesity and metabolic biomarkers in Brazilian individuals / Associação entre variantes do gene de leptina e obesidade e biomarcadores metabólicos em indivíduos brasileiros

OBJECTIVE: The relationship between variants of the leptin gene (LEP) and obesity and metabolic biomarkers was investigated in Brazilian individuals. SUBJECTS AND METHODS: One-hundred-ten obese (BMI > 30 kg/m²) and 100 non-obese individuals (145 women and 65 men, aged 49 ± 14 years) were randomly selected. Plasma leptin, glycemia, serum lipid measurements and LEP -2548G>A and 3'HVR polymorphisms were analyzed. RESULTS: The LEP -2548GG genotype was associated with a 2.2 percent and 2.0 percent increase in BMI (p = 0.009) and plasma leptin (p = 0.031), respectively. 3'HVR I/II (classes I/I+I/II) genotypes contributed with 1.8 percent of BMI values (p = 0.046). LEP I/G combined genotypes (I/IGG, I/IGA and I/IIGG) were associated with obesity, and increased BMI, waist circumference, leptin and triglycerides (p < 0.05). These relationships were found in women (p < 0.05) but not in men. LEP I/G combined genotypes were not associated with hypertension, hyperglycemia, dyslipidemia and coronary artery disease. CONCLUSIONS: LEP I/G combined genotypes are associated with obesity-related metabolic biomarkers and phenotype in a gender-dependent manner.

OBJETIVO: A relação entre as variantes do gene da leptina (LEP) e obesidade e biomarcadores metabólicos foi investigada em indivíduos brasileiros. SUJEITOS E MÉTOODS: Cento e dez indivíduos obesos (IMC > 30 kg/m²) e 100 não obesos (145 mulheres e 65 homens, idade 49 ± 14 anos) foram selecionados aleatoriamente. Leptina plasmática, glicemia, lípides séricos e polimorfismos LEP -2548G>A e 3'HVR foram analisados. RESULTADOS: O genótipo -2548GG foi associado com aumento de 2,2 por cento e 2,0 por cento no IMC (p = 0,009) e leptina plasmática (p = 0,031), respectivamente, enquanto os genótipos 3´HVR I/II (classes I/I+I/II) contribuíram com 1,8 por cento dos valores de IMC (p = 0,046). Os genótipos combinados LEP I/G (I/IGG, I/IGA e I/IIGG) foram associados com obesidade e IMC aumentado, circunferência abdominal, leptina e triglicérides aumentados (p < 0,05). Essas relações foram encontradas em mulheres (p < 0,05), mas não em homens. Os genótipos LEP I/G combinados não foram associados com hipertensão, hiperglicemia, dislipidemia e doença arterial coronariana. CONCLUSÕES: Genótipos combinados LEP I/G são associados com biomarcadores metabólicos e fenótipo de obesidade de forma gênero-dependente.

Leptin G-2548A promoter polymorphism is associated with increased plasma leptin and BMI in Brazilian women / Polimorfismo do promotor do gene da leptina está associado ao aumento de leptina plasmática e IMC em mulheres brasileiras

Variants in leptin gene (LEP) have been implicated in the pathogenesis of obesity. The relationship between LEP G-2548A polymorphism and obesity-related traits was evaluated in a sample of Brazilian women (n = 228) who were randomly selected from two clinical centers in Sao Paulo city. Blood samples were collected for DNA extraction, plasma leptin and serum lipids measurements. LEP G-2548A genotypes were identified by a PCR- RFLP strategy using the endonuclease Alw44I. LEP G-2548A was associated with obesity after adjustment for covariates (age, hypertension, coronary artery disease, smoking and physical activity). Women carrying G allele had a four times higher risk of obesity than the A allele carriers (OR: 4.11, CI95 percent: 1.06-15.90, p = 0.041). G allele was also related to increased plasma leptin (p = 0.024) and body mass index (p = 0.027). Hypertension, hyperglycemia, dyslipidemia and coronary artery disease were associated with obesity. However LEP G-2548A polymorphism was not related to these variables. All together these data suggest that LEP G-2548A polymorphism has an important role in regulating plasma leptin levels and body mass index in women.

Variantes no gene da leptina (LEP) foram implicados na patogênese da obesidade. A relação entre o polimorfismo LEP G-2548A e as características relacionadas com a obesidade foram avaliadas em mulheres brasileiras (n = 228), que foram selecionadas randomicamente de dois centros de pesquisa clínica na cidade de São Paulo. As amostras de sangue foram coletadas para extração de DNA e determinações de leptina plasmática e lipídeos séricos. Os genótipos do LEP G-2548A foram identificados pela estratégia de PCR-RFLP, empregando a endonuclease Alw44I. O polimorfismo LEP G-2548A foi associado com obesidade, após ajuste para as covariáveis: idade, hipertensão, doença arterial coronariana, tabagismo e atividade física. Mulheres com alelo G tiveram quatro vezes maior risco de obesidade que as portadoras do alelo A (OR: 4,11, CI95 por cento: 1,06-15,90; p = 0,041). O alelo G também foi relacionado com leptina plasmática (p = 0,024) e o índice de massa corporal (p = 0,027) aumentado. A hipertensão, a hiperglicemia, a dislipidemia e a doença arterial coronariana foram associadas com obesidade. Entretanto, o polimorfismo LEP G-2548A não foi relacionado com essas variáveis. Os resultados deste estudo são sugestivos de que o polimorfismo LEP G-2548A tem papel importante na regulação da leptina plasmática e no índice de massa corporal em mulheres.

Pseudoaneurisma embolomicótico da artéria femoral comum na endocardite infecciosa / Embolomycotic pseudoaneurysm of the commom femoral artery in infective endocarditis

Paciente do sexo masculino, seis anos de idade, com quadro clínico e ecocardiográfico compátivel com endocardite infecciosa de valva mitral, apresentou como complicaçäo, pseudoaneurisma embolomicótico da artéria femoral comum direita. Submetido a tratamento cirúrgico com a reconstruçäo arterial, evoluiu satisfatoriamente

Terapêutica antiisquêmica: bloqueadores beta-adrenérgicos, nitratos e antagonistas dos canais de cálcio / Anti-ischemic therapy: beta-blockers, nitrates, calcium antagonists

Os medicamentos antiisquêmicos são a base, a seqüência e a continuidade terapêutica dos pacientes portadores de angina estável. Vários estudos comprovaram que os bloqueadores beta-adrenérgicos melhoram a qualidade de vida e aumentam a sobrevida dos pacientes com angina estável e com antecedentes de infarto do miocárdio. Os nitratos, medicamentos pioneiros no tratamento da angina estável, apesar de pouco estudados, têm sido eficientes na redução e no controle das crises de angina. Os benefícios dos antagonistas dos canais de cálcio na prevenção e no tratamento das crises de angina somam-se a seus efeitos nas co-morbidades que feqüentemente se associam com as síndromes coronárias crônicas.