Perfil peptídico de hidrolisados enzimáticos do concentrado proteico do soro de leite, obtidos pela ação da pancreatina e da papaína / Peptide profile of enzymatic hydrolysates from whey protein concentrate obtained by action of pancreatin and papain

The aim of this study was to obtain enzymatic hydrolysates from whey protein concentrate with high oligopeptide, especially di- and tripeptides, and free amino acid contents, besides small amounts of large peptides. Different parameters were evaluated such as type of enzyme (pancreatin and papain), enzyme:substrate ratio (0.5:100, 1:100, 2:100 and 3:100), and the use of ultrafi ltration. The peptide profi les of the hydrolysates were characterized by using a fractionation method by size-exclusion HPLC followed by a rapid Corrected Fraction Area method for quantifying the components of the chromatographic fractions. The results showed that, in terms of number of analyzed samples, the pancreatin action was more advantageous than papain. However, the best peptide profile was obtained by papain, reaching 15.29% of di- and tripeptides, 47.83% of free amino acids and 25.73% of large peptides. The use of the smallest enzyme:substrate ratio (0.5:100) was beneficial in some cases for both enzymes, while the lack of ultrafiltration was favorable just for pancreatin.

El objetivo de este trabajo fue obtener hidrolizados enzimáticos a partir de concentrado proteico de suero lácteo con elevados contenidos de oligopéptidos, especialmente di y tri péptidos, y aminoácidos libres, además de reducida cantidad de grandes péptidos. Para eso fueron evaluados algunos parámetros como el tipo de enzima (pancreatina o papaína), la relación enzima:substrato (0,5:100, 1:100, 2:100 y 3:100)y el empleo de ultrafiltración. Para evaluar el perfil peptídico, los hidrolizados se sometieron a fraccionamiento en cromatografía líquida de alta eficiencia de exclusión molecular (SE-HPLC) y para la cuantificación de los componentes de las fracciones de la cromatografía fue utilizado el método del Área Corregida de la Fracción (ACF). Los resultados indican que el uso de pancreatina fue más ventajoso que el de papaína, así como la menor relación enzima:substrato (0,5:100). Por otro lado, el mejor perfil peptídico (15,29% de di y tripéptidos, 47,83% de aminoácidos libresy 25,73% de grandes péptidos) fue conseguido con el empleo de papaína. La menor relación enzima:substrato (0,5:100) fue favorable en algunos casos para las dos enzimas. La ausencia de ultrafiltración favoreció los hidrolizados producidos con pancreatina.

O presente trabalho teve como objetivo a obtenção de hidrolisados enzimáticos do concentrado proteico do soro de leite com elevado teor de oligopeptídeos, principalmente di- e tripeptídeos, e de aminoácidos livres, além de quantidade reduzida de grandes peptídeos. Para tal, foram avaliados diferentes parâmetros, como tipo de enzima (pancreatina e papaína), relação enzima:substrato (0,5:100, 1:100, 2:100 e 3:100) e o emprego da ultrafiltração. Caracterizou-se o perfil peptídico pelo fracionamento dos hidrolisados por cromatografia líquida de alta eficiência de exclusão molecular e, para a quantificação dos componentes das frações cromatográficas, empregou-se o método rápido da Área Corrigida da Fração. Os resultados obtidos indicaram que, em termos de número de casos analisados, a ação da pancreatina foi mais vantajosa do que a da papaína. Entretanto, o melhor perfil peptídico foi obtido pela açãoda papaína, dando origem a 15,29% de di- e tripeptídeos, 47,83% de aminoácidos livres e 25,73% de grandes peptídeos. A utilização da menor relação enzima:substrato (0,5:100) foi benéfica em alguns casos para ambas as enzimas, enquanto que a ausência da ultrafiltração mostrou-se favorável apenas para a pancreatina.

Study of slow ultrafilteration as a therapeutic modality for chronic refractory congestive heart failure

Refractory heart failure usually represents the end stage of congestive heart failure in which hypotension and oliguria, lead to progressive generalized edema. The purpose of this study was to assess the efficacy of extracorporeal ultrafilteration [ECUF] as a modality for treatment of patients with refractory congestive heart failure who failed to respond to conventional anti failure measures. The study was carried out at a period for about six months and it included 40 patients, 18 men [45%] and 22 women [55%], they were selected from patients admitted to the intensive care of Internal Medicine Department and cardiology Department of Tanta University Hospital. Selection criteria were NYHA class III and IV heart failure, resting left ventricular ejection fraction [LVEF] <35, normal or mild elevated serum creatinine [patients with chronic renal failure were excluded from the study], lack of satisfactory response to conventional therapeutic regimen. All patients were subjected To ECUF ranged from 3 sessions in 12 patients and 4 sessions in 28 patients [mean 3.7 session/ptn] and between [4-6 hour/session]. It was initated at a rate of 0.5L/h to be readjusted subsequently according to the hemodynamic parameters available. Average amount of ultrafilterate/session ranged from 2.5-3 liters [mean2.6 +/- 0.4]. Both number and repition frequency were decided empirically on the basis of evolution of symptoms and response to drugs. All patients were subjected to the following assessment before and after ECUF: Daily morning checking of body weight [kg], hematocrit value [HTC%], urin output [ml/24h], serum electrolytes [Na[+], K[+]], blood urea and serum creatinine, hemodynamic monitoring including [heart rate, blood pressure and CVP], chest x-ray P-A view for measurment of CTR, E.C.G and transthoracic echocardiography for evaluation of cardiac function. Our results showed that there was significant increase of diuresis from [0.3410.14 to 2.0 +/- 0.7L/day] [p<0.001], significant reduction of mean H.R [120+5.0 to 98 +/- 11.0 beat/min] [p<0.001], significant CVP reduction [28.5 +/- 2.o to 15.5 +/- 7.0 cm water] [p<0.001], decrease of intravenous volume as assessed by significant increase of HCT value from [33.8 +/- 2.3% to 37.6 +/- 2.0%] [p<0.001] after ECUF, significant decrease of S. urea from [105.7 +/- 55.2 to 91.3 +/- 63.o4 mmol/L] and S. creatinine from [1.9 +/- 0.6 to 1.5 +/- 0.9 mmol/L] after ECUF. ECHO repeated after ECUF revealed highly significant reduction in the mean LVEDD from [7.0 +/- 0.2 cm to 6.4 +/- 0.6 cm] [p<0.001], reduction of LVESD from [6.5 +/- 0.4 cm to 5.5 +/- 0.8 cm] [p<0.001], reduction in mean LA dimension from [5.3 +/- 0.3 cm to 4.8 +/- 0.6 cm] [p<0.001], reduction of RV dimension from [4.7 +/- 0.4 cm to 4.0 +/- 0.8 cm] [p<0.001], significant increase of LVEF from [23.8 +/- 4.1 to 31.2 +/- 8.2] [p<0.001]: ECUF offers a reasonable effective and relatively safe method in the treatment of refractory heart failure. The relative ease of the procedure and low incidence of complications denote that this technique is a useful one in selected cases of heart failure, refractory to the conventional therapy with oliguria and fluid overload