Human Tissue Kallikrein Activity in Angiographically Documented Chronic Stable Coronary Artery Disease / Atividade da Calicreína Tecidual Humana na Doença Coronariana Crônica Estável Angiograficamente Documentada

AbstractBackground:Human tissue kallikrein (hK1) is a key enzyme in the kallikrein–kinin system (KKS). hK1-specific amidase activity is reduced in urine samples from hypertensive and heart failure (HF) patients. The pathophysiologic role of hK1 in coronary artery disease (CAD) remains unclear.Objective:To evaluate hK1-specific amidase activity in the urine of CAD patientsMethods:Sixty-five individuals (18–75 years) who underwent cardiac catheterism (CATH) were included. Random midstream urine samples were collected immediately before CATH. Patients were classified in two groups according to the presence of coronary lesions: CAD (43 patients) and non-CAD (22 patients). hK1 amidase activity was estimated using the chromogenic substrate D-Val-Leu-Arg-Nan. Creatinine was determined using Jaffé’s method. Urinary hK1-specific amidase activity was expressed as µM/(min · mg creatinine) to correct for differences in urine flow rates.Results:Urinary hK1-specific amidase activity levels were similar between CAD [0.146 µM/(min ·mg creatinine)] and non-CAD [0.189 µM/(min . mg creatinine)] patients (p = 0.803) and remained similar to values previously reported for hypertensive patients [0.210 µM/(min . mg creatinine)] and HF patients [0.104 µM/(min . mg creatinine)]. CAD severity and hypertension were not observed to significantly affect urinary hK1-specific amidase activity.Conclusion:CAD patients had low levels of urinary hK1-specific amidase activity, suggesting that renal KKS activity may be reduced in patients with this disease.

ResumoFundamento:A calicreína tecidual humana (hK1) é enzima-chave do sistema calicreína-cinina (SCC). A atividade amidásica da hK1 está reduzida na urina de pacientes com hipertensão e insuficiência cardíaca (IC); seu papel na doença arterial (DAC) coronariana ainda não está esclarecido.Objetivo:Avaliar a atividade amidásica da hK1 na urina de pacientes com DAC.Métodos:Sessenta e cinco indivíduos (18 a 75 anos) que se submeteram ao cateterismo cardíaco (CAT) coletaram amostra do jato médio de urina imediatamente antes do CAT. Baseando-se na presença de lesões coronarianas, os pacientes eram classificados em dois grupos: DAC (43 pacientes) e sem DAC (22 indivíduos). A atividade amidásica da hK1 foi estimada com o substrato cromogênico D-Val-Leu-Arg-Nan. Creatinina foi determinada pelo método de Jaffé. A atividade amidásica específica da hK1 urinária foi expressa em µM/(min . mg de creatinina) para corrigir diferenças no fluxo urinário.Resultados:A atividade amidásica da hK1 urinária foi semelhante entre os pacientes com DAC [0,146 µM/(min . mg de creatinina)] e aqueles sem DAC [0,189 µM/(min . mg de creatinina)] (p = 0,803), e permaneceu entre os baixos valores previamente publicados para pacientes com hipertensão primária [0,210 µM/(min . mg de creatinina)] e para aqueles com IC [0,104 µM/(min . mg de creatinina)], respectivamente. Nenhum efeito estatisticamente significativo da gravidade da DAC e da hipertensão sobre a atividade amidásica da hK1 urinária foi observado.Conclusão:A atividade amidásica da hK1 na urina estava reduzida nos pacientes com DAC, o que pode sugerir que a atividade do SCC renal esteja reduzida nessa doença.

Human Tissue Kallikrein Activity in Angiographically Documented Chronic Stable Coronary Artery Disease.

BACKGROUND: Human tissue kallikrein (hK1) is a key enzyme in the kallikrein-kinin system (KKS). hK1-specific amidase activity is reduced in urine samples from hypertensive and heart failure (HF) patients. The pathophysiologic role of hK1 in coronary artery disease (CAD) remains unclear. OBJECTIVE: To evaluate hK1-specific amidase activity in the urine of CAD patientsMethods:Sixty-five individuals (18-75 years) who underwent cardiac catheterism (CATH) were included. Random midstream urine samples were collected immediately before CATH. Patients were classified in two groups according to the presence of coronary lesions: CAD (43 patients) and non-CAD (22 patients). hK1 amidase activity was estimated using the chromogenic substrate D-Val-Leu-Arg-Nan. Creatinine was determined using Jaffé's method. Urinary hK1-specific amidase activity was expressed as µM/(min · mg creatinine) to correct for differences in urine flow rates. RESULTS: Urinary hK1-specific amidase activity levels were similar between CAD [0.146 µM/(min ·mg creatinine)] and non-CAD [0.189 µM/(min . mg creatinine)] patients (p = 0.803) and remained similar to values previously reported for hypertensive patients [0.210 µM/(min . mg creatinine)] and HF patients [0.104 µM/(min . mg creatinine)]. CAD severity and hypertension were not observed to significantly affect urinary hK1-specific amidase activity. CONCLUSION: CAD patients had low levels of urinary hK1-specific amidase activity, suggesting that renal KKS activity may be reduced in patients with this disease.

Increased tissue kallikrein amidase activity in urine of patients with type 1 diabetes under insulin therapy, and in those with gestational diabetes mellitus not under insulin therapy.

Human tissue kallikrein (hK1) is reduced in hypertension, cardiovascular and renal diseases. There is little information on the participation of hK1 in type 1 diabetes mellitus (DM), type 2 DM, and gestational diabetes mellitus (GDM), respectively. The aim of this study was to evaluate the roles of insulin and hyperglycemia on urinary hK1 activity in type 1 DM and in GDM. Forty-three type 1 DM patients (5-35 years, disease duration ≤ 5years, receiving insulin, HbA(1c)>7.6%) were selected. Forty-three healthy individuals, paired according to gender and age, were used as controls. Thirty GDM patients (18-42 years, between the 24th and 37th week of pregnancy, recently diagnosed, not under insulin therapy) were also selected. Thirty healthy pregnant (18-42years, between the 24th and 37th week of pregnancy) and 30 healthy non-pregnant women (18-42years) were selected as controls. Random midstream urine was used. hK1 amidase activity was estimated with D-Val-Leu-Arg-Nan substrate. Creatinine was determined by Jaffe's method. hK1 specific amidase activity was expressed as µM/(minmg creatinine) to correct for differences in urine flow rate. hK1 specific amidase activity was significantly higher in the urine of type 1 DM than in controls, and in the urine of GDM patients than in healthy pregnant women and healthy non-pregnant women, respectively. The data suggest that hyperglycemia, rather than insulin, is involved in the mechanism of increased hK1 specific amidase activity in both type 1 DM and GDM patients, respectively.

On human tissue kallikrein activity in urine of Brazilian White and Black primary hypertensive patients.

OBJECTIVE: To evaluate the possible relationship between the human tissue kallikrein amidase activity with sex and ethnicity in Brazilian primary hypertensive patients. DESIGN: Population-based study. PARTICIPANTS: One hundred men and women, Black and White primary hypertensive patients aged 20 years and older were selected. Eighty nine healthy individuals, paired according to age, sex, and ethnics were used as controls. METHODS: Early-morning midstream urine was used. Human tissue kallikrein amidase activity was estimated with D-Val-Leu-Arg 4-nitroanilide substrate. Creatinine was determined by a method based on Jaffe's reaction. hK1 amidase activity is expressed in microM/(min x mg creatinine) to correct for differences in urine flow rate. Data are expressed as medians. RESULTS: Human tissue kallikrein amidase activity was significantly lower in the urine of hypertensive patients (0.210 microM/(min x mg creatinine) than in the urine of control subjects (0.260 microM/(min x mg creatinine) (P = .010). This result supports data from the literature. Contrasting to what was already reported, namely that human tissue kallikrein excretion is higher in females than in males, and especially higher in Caucasians than in African Americans, our results show that, in the urine of Brazilian hypertensive patients and control subjects, no significant effect of sex and ethnicity on human tissue kallikrein amidase activity was observed. CONCLUSIONS: The lack of ethnicity effect supports what was already asserted, namely, that, in Brazil, at an individual level, color, as determined by physical evaluation, is a poor predictor of genomic African ancestry, estimated by molecular markers.

Perfil peptídico de hidrolisados enzimáticos de leite em pó desnatado / Peptide profile of enzymatic hydrolysates from skim milk powder

Sete hidrolisados de leite em pó desnatado foram preparados, visando à produção de hidrolisados protéicos, como suplemento dietético para fenilcetonúricos. Para isso, utilizaram-se uma protease do Aspergillus oryzae (AO), a papaína (PA) e a pepsina (PE), isoladamente ou não em associação, em diferentes relações enzima:substrato (E:S), sendo o tempo total de reação de 5 h e a temperatura de 50°C. Adotou-se com critério para a avaliação nutricional dos hidrolisados, o estudo da distribuição dos peptídeos nas amostras, de acordo com o tamanho da cadeia. Inicialmente, os hidrolisados foram fracionados por cromatografia líquida de alta eficiência de exclusão molecular (SE-HPLC) e, para a quantificação dos componentes das frações cromatográficas, empregou-se o método rápido da Área Corrigida da Fração (ACF)...

Hidrolisados enzimáticos de leite em pó desnatado como fonte de oligopeptídeos para formulações dietéticas / Enzymatic hydrolysates from skimmed powder milk as source of oligopeptides for dietary formulations

Seis hidrolisados de leite em pó desnatado foram preparados visando a produção de hidrolisados protéicos para uso em formulações dietéticas para fenilcetonúricos. Para isso utilizaram-se uma protease do Aspergillus oryzae (AO), isoladamente ou em associação com a papaína (PA), em diferentes tempos de reação e relação enzima:substrato (E:S). O interesse deste trabalho está associado ao estudo da distribuição dos peptídios nestas amostras, de acordo com o tamanho da cadeia, como critério de avaliação de sua qualidade nutricional. Inicialmente, os hidrolisados foram fracionados por cromatografia líquida de alta eficiência de exclusão molecular (SE-HPLC) e, para a quantificação dos componentes das frações cromatográficas, empregou-se o método rápido da Área Corrigida da Fração (ACF)...