INTRAMOLECULARLY QUENCHED FLUOROGENIC PEPTIDE-SUBSTRATES for HUMAN RENIN

UNIV FED MINAS GERAIS,DEPT BIOCHEM & IMMUNOL,BR-30000 BELO HORIZONTE,MG,BRAZILWeb of Scienc

Kinetic characterization of rat tissue kallikrein using N alpha-substituted arginine 4-nitroanilides and N alpha-benzoyl-L-arginine ethyl ester as substrates

Hydrolysis of seven Nor-substituted L-arginine 4-nitroanilides: benzoyl-arginine p-nitroanilide (Bz-Arg-Nan), tosyl-arginine p-nitroanilide (Tos-Arg-Nan), acetyl-leucyl-arginine p-nitroanilide (Ac-Leu-Arg-Nan), acetyl-phenylalanyl-arginine p-nitroanilide (Ac-Phe-Arg-Nan), benzoyl-phenylalanyl-arginine p-nitroanilide (Bz-Phe-Arg-Nan), tosyl-phenylalanyl-arginine p-nitroanilide (Tos-Phe-Arg-Nan), and D-valyl-leucyl-aginine p-nitroanilide (D-Val-Leu-Arg-Nan), and the N alpha-substituted L-arginine ester: benzoyl-arginine ethyl ester (Bz-Arg-OEt), by rat tissue kallikrein was studied throughout a wide range of substrate concentrations, The enzyme showed a bimodal behavior with all the substrates tested except Tos-Arg-Nan, At low substrate concentrations (10 to 170 mu M for p-nitroanilides and 50 to 190 mu M for Bz-Arg-OEt) the hydrolysis followed Michaelis-Menten kinetics, but at higher substrate concentrations (150 to 700 mu M for p-nitroanilides and 200 to 1800 mu M for Bz-Arg-OEt) a deviation from Michaelis-Menten kinetics was observed with a significant decrease in hydrolysis rates. At high concentrations of the p-nitroanilide substrates, partial enzyme inhibition was observed, whereas complete enzyme inhibition was observed with Bz-Arg-OEt at high concentration, The kinetic parameters reported here were calculated from data for substrate concentrations range where the enzyme followed Michaelis-Menten behavior. D-Val-Leu-Arg-Nan (K-m = 24 +/- 2 mu M; V-max = 10.42 +/- 0.28 mu M/min) was the best substrate tested, followed by Ac-Phe-Arg-Nan (K-m = 13 +/- 2 mu M; V-max = 3.21 +/- 0.11 mu M/min), while Tos-Arg-Nan (K-m = 29 +/- 2 mu M; V-max = 0.10 +/- 0.002 mu M/min) was the worst of the tested substrates for rat tissue kallikrein, For the hydrolysis of Bz-Arg-OEt (K-m = 125 +/- 15 mu M; V-max = 121.3 +/- 7.6 mu M/min), the kinetic parameters using a substrate inhibition model can reasonably account for the observed enzyme behavior, with a K-si value about 13.6 times larger than the estimated K-m value.UNIV FED MINAS GERAIS,FAC FARM,DEPT ANAL CLIN & TOXICOL,BR-30150112 BELO HORIZONT,MG,BRAZILUNIV FED MINAS GERAIS,INST CIENCIAS BIOL,DEPT BIOQUIM & IMUNOL,BR-30160042 BELO HORIZONT,MG,BRAZILUNIV FED OURO PRETO,FAC FARM,DEPT FARM,BR-35400000 OURO PRETO,MG,BRAZILUNIV FED SAO PAULO,DEPT BIOFIS,BR-04023062 SAO PAULO,BRAZILUNIV FED SAO PAULO,DEPT BIOFIS,BR-04023062 SAO PAULO,BRAZILWeb of Scienc

Validação inicial do índice de necessidade de atenção à saúde bucal para as equipes de saúde bucal na estratégia de saúde da família Initial validation of the index of oral healtcare needs for oral health teams in the family healthcare strategy

Este trabalho objetivou validar o Índice de Necessidade de Atenção à Saúde Bucal (INASB), a partir de um algoritmo pré-definido baseado nas condições sociais das famílias. Utilizaram-se informações obtidas na ficha A do sistema de informação da atenção básica. A validação do índice foi realizada através da validação de face e de construto, e nesta foram coletados dados de experiência de cárie, dor de dente e acesso aos serviços de saúde bucal através de uma amostra aleatória, estratificada e baseada no índice de 412 crianças nas faixas etárias 3-5 e 7-12 anos em Recife-PE. Para análise foi utilizado o nível de significância de 5%. O índice foi considerado adequado na validação de face. Na validação de construto convergente foi associado ao componente cariado do ceo-d (p = 0,03) e CPO-D (p = 0,01); e na divergente foi associado ao acesso ao dentista (p = 0,001) e ao componente obturado do ceo-d (p = 0,05) não sendo associado ao componente obturado do CPO-D. O Índice de Necessidade de Atenção à Saúde Bucal demonstrou possuir uma boa validade inicial podendo ser usado como instrumento de programação para as equipes de saúde bucal da família.<br>This survey set out to validate the Index of Oral Healthcare Needs (IOHN), based on a pre-defined algorithm of the social status of families. The validation process was divided into two phases, namely a face validation and a construct validation. In the latter, data on caries experience, toothache and access to oral health services was collected. To validate the index a random, stratified sample of 412 children aged 3-5 and 7-12 was obtained, based on the IOHCN algorithm, all the children being from the areas of Recife covered by the family healthcare program. The analysis consisted of a descriptive and an analytical phase, adopting a 5% level of significance. The index was considered by an expert committee to have good face validity. The convergent construct validation was associated with a decay component of dmft (p = 0.03) and DMFT (p = 0.01); the divergent construct validation was associated with access to oral care (p = 0.001) and filled component of dmft (p = 0.05), showing no association with the filled component of DMFT. The Index of Oral Healthcare Needs was shown to have good initial validation and canbe used as a useful tool in the planning of dental care at a local level

Alleviation of Abiotic and Biotic Stresses in Plants by Azospirillum

International audienc