Pharmacogenetic implications in the management of metabolic diseases in Brazilian populations

Dyslipidemia, diabetes, obesity and hypertension are common metabolic diseases. In the last decades, unhealthy lifestyle and aging have leads to an increased incidence of these diseases, increasing morbidity and mortality by cardiovascular causes. The treatment of metabolic diseases includes lifestyle interventions as healthy diet and physical exercise, as well as pharmacological interventions. Several drugs are available for the management of metabolic diseases including among others lipidlowering antidiabetics and antihypertensive drugs. Variability in response to these drugs is influenced by both genetic and non-genetic factors. Polymorphisms in genes related to drug pharmacokinetics and pharmacodynamics have been shown to influence drug efficacy and safety. This review is focused on pharmacogenetic studies related to the management of metabolic diseases in samples of the Brazilian population. Associations of variants in drug metabolizing enzymes and transporters, drug target and metabolism-related genes with the efficacy and safety of lipid-lowering, antidiabetic and antihypertensive drugs are described. Most pharmacogenetic studies in Brazil have focused in pharmacological response to a small group of drugs, as statins and some antihypertensives, while there are almost no studies on antidiabetic and antiobesity drugs. Some studies reported significant associations of gene polymorphisms with drug response confirming previous data from other populations, whereas other works did not replicate, which may relay on the genetic admixture of our population. In conclusion, further studies are necessary considering larger sample sizes, new unexplored drugs and more genetic variants to obtain stronger conclusions to explore clinical applications of pharmacogenetic studies in our population

FARMACOGENÉTICA DO TRATAMENTO DE DIABETE MELITO

Diabetes mellitus is a metabolic disease involving the deficiency or resistance to insulin that triggers to severe micro and macrovascular complications. Both genetic and environmental factors contribute to the complex etiology of main forms of diabetes. The recent advances in molecular genetics contributed to the identification of several loci and candidate genes that are related to the clinically recognized diabetes subtypes. This knowledge has become more important for recognition of diabetic patients with differential response to specific hypoglycemic drugs. In this study, the molecular aspects of the monogenic and polygenic forms of diabetes as well as the pharmacogenetics approach of the drugs commonly used in treatment are reviewed.Diabete melito é uma doença metabólica caracterizada pela deficiência ou resistência de insulina que leva a complicações micro e macrovasculares graves. Fatores genéticos e ambientais contribuem para a etiologia complexa das principais formas de diabete. Os avanços recentes na genética molecular contribuíram para a identificação de vários locos e genes que são relacionados aos subtipos de diabete clinicamente reconhecidos. Este conhecimento tornou-se muito importante para o reconhecimento de pacientes diabéticos com resposta diferencial a fármacos hipoglicemiantes específicos. Neste estudo serão revisados os aspectos moleculares das formas monogênicas e poligênicas da diabete assim como a abordagem farmacogenética dos agentes terapêuticos mais comuns

Pitfalls in DNA extraction from ancient bones found in Brazilian shell-mounds

Os autores descrevem suas dificuldades na extração de ácido desoxiribonuclêico (DNA) de ossos encontrados em sambaquis nas praias da ilha de Sta. Carina, SC, Brasil. A presença de ácido húmico, do ácido fúlvico, radicais livres oxidantes e o efeito hidrolítico dos ambientes úmidos degradam o DNA em fragmentos da ordem de 20-30 pares de bases, impedindo qualquer utilização do DNA extraído. Além disso, o ácido húmico e o ácido fúlvico se ligam fortemente ao magnésio, necessário à atividade da Taq-polimerase, impedindo a reação da polimerase em cadeia (PCR). Neste trabalho são discutidos os protocolos utilizados mais eficientes de extração do DNA e de várias condições possíveis de ensaio da PCR.The authors report their difficulties in DNA extraction from ancient bones found in shell-mounds of Sta. Catarina Island, SC, Brazil. The presence of humic acid, fulvic acid, free oxidant radicals and the hydrolytic effect of wet environment degrade the DNA to 20 to 30 base pairs, what jeopardizes any further utilization. The humic acid and the fulvic acid chelate magnesium ion necessary to Taq-polymerase activity, inhibiting the polymerase chain reaction (PCR). In this work the most efficient used protocols for DNA extraction, and the several PCR assay conditions are discussed

Exposure to potentially inappropriate medications in Brazilian elderly outpatients with metabolic diseases

Management of pharmacotherapy in elderly with metabolic diseases is challenging and potentially inappropriate medications (PIMs) are risk factors for drug interactions and adverse events. The exposure to PIMs in elderly outpatients with metabolic diseases and its relationship with polypharmacy and other variables was investigated. PIMs prescribed to 207 elderly patients (aged 60 to 96 years) with metabolic diseases who attended a University Hospital of Sao Paulo city, Brazil, from April/2010 to January/2011, were evaluated. PIMs were detected using both 2003 Beers and 2008 STOPP criteria. The association between PIMs and age, gender and polypharmacy was also examined. 2008 STOPP criteria detected more PIMs (44.4 %) than 2003 Beers criteria (16.0%,

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% 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.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% e 2,0% 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% 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

Application of BRED technology to construct recombinant D29 reporter phage expressing EGFP

Bacteriophage Recombineering of Electroporated DNA (BRED) has been described for construction of gene deletion and point mutations in mycobacteriophages. Using BRED, we inserted a Phsp60-egfp cassette (1143 bp) into the mycobacteriophage D29 genome to construct a new reporter phage, which was used for detection of mycobacterial cells. The cassette was successfully inserted and recombinant mycobacteriophage purified. DNA sequencing of the cassette did not show any mutations even after several phage generations. Mycobacterium smegmatis mc2155 cells were infected with D29::Phsp60-egfp (MOI of 10) and evaluated for EGFP expression by microscopy. Fluorescence was observed at around 2 h after infection, but dissipated in later times because of cell lysis. We attempted to construct a lysis-defective mutant by deleting the lysA gene, although we were unable to purify the mutant to homogeneity even with complementation. These observations demonstrate the ability of BRED to insert c. 1 kbp-sized DNA segments into mycobacteriophage genomes as a strategy for constructing new diagnostic reporter phages.Fil: Silva, Joas L. da. Universidade de Sao Paulo; BrasilFil: Piuri, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Broussard, Gregory. University Of Pittsburgh; Estados UnidosFil: Marinelli, Laura J.. University Of Pittsburgh; Estados UnidosFil: Bastos, Gisele M.. Universidade de Sao Paulo; BrasilFil: Hirata, Rosario D. C.. Universidade de Sao Paulo; BrasilFil: Hatfull, Graham F.. University Of Pittsburgh; Estados UnidosFil: Hirata, Mario H.. Universidade de Sao Paulo; Brasi

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%: 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%: 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