Saline as solvent and ethanol-based purification process for the extraction of proteins and isoflavones from wet okara.

Okara, the solid byproduct of water-soluble soybean extract production, is a potential source of proteins and isoflavones. This study investigated different experimental configurations for extracting these compounds from wet okara, including lipid removal with ethanol at different stages of the recovery process, sequential crosscurrent extraction, and using a saline MgCl2 solution as the solvent. Three washes with a 60:40 ethanol:water (w/w) solution after isoelectric precipitation significantly increased protein content by reducing lipid content (60 %). The crosscurrent approach using 0.05 M MgCl2 yielded okara proteinaceous material with 248 µg/g daidzein and 236 µg/g genistein, along with a 3 % increase in protein content, attributed to enhanced extraction of 7S globulins. These configurations notably affected the functional properties of the protein materials. Overall, this research provides detailed insights into the composition and properties of proteins extracted from wet okara, facilitating their specialized application in food products.

Effect of different selenium sources and concentrations on glutathione peroxidase activity and cholesterol metabolism of beef cattle.

The objective of this study was to investigate the effects of different Se sources and concentrations on glutathione forms and cholesterol metabolism in beef cattle. Sixty-three Nellore bulls (412 ± 19 kg body weight (BW); 24 mo old) were randomly assigned to a completely randomized design in a 2 × 3 + 1 factorial arrangement (63 pens; one animal/pen) with two Se sources (sodium selenite, ING and Se-yeast, ORG), three concentrations (0.3, 0.9, and 2.7 mg supplemental Se/kg dry matter (DM)), and control treatment (without Se supplementation) fed for 90 d. Blood samples were collected on day 0, 28, 56, and 84. Muscle and liver samples were collected at harvest. Hepatic GSSG (P = 0.004), GSH/GSSG ratio (P = 0.030), and GSH-Px (P = 0.004) were affected by Se source × concentration interaction. Oxidized glutathione was higher in the ORG group vs. ING at concentration 2.7 mg supplemental Se/kg DM, but at 0.3 mg supplemental Se/kg DM the ING group was higher than ORG. The liver GSH-Px activity was higher in the ORG group vs. ING at concentration 0.9 and 2.7 mg supplemental Se/kg DM. The GSH/GSSG ratio was the highest in animals fed 0.3 mg supplemental Se/kg DM of ORG. Selenium liver concentration increased linearly with the supplemental Se concentration in the diet (y = 0.0583 + 0.4254x, R2 = 0.92, P < 0.0001), regardless of source. Total meat cholesterol was greater (P < 0.001) in CON (control) vs. SUP (supplemented, regardless source) group. The muscle GSH-Px activity was higher (P < 0.001) in SUP vs. CON and increased (P < 0.004) with increasing supplemental Se concentrations. There was an increase on very low-density lipoprotein (VLDL), glucose, and triglycerides in ORG vs. ING (P ≤ 0.035). In general, serum Se was higher (P < 0.001) in SUP vs. CON and increased with increasing supplemental Se concentration. Lastly, the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR) concentration was lower (P = 0.002) in SUP (0.39 ng/mL) vs. CON (0.55 ng/mL). Selenium supplementation with different sources and concentrations has the potential to affect cholesterol metabolism by affecting GSH/GSSG ratio, GSH-Px, and the HMGCR.

Protective action of indole-3-acetic acid on induced hepatocarcinoma in mice.

In this study, we report the protective effects of IAA on diethylnitrosamine (DEN)-induced hepatocarcinogenesis. BALB/c mice received daily IAA at 50 (T(50)), 250 (T(250)), and 500 (T(500)) mg Kg(-1) per body mass by gavage for 15 days. At day 15, animals were administered DEN and sacrificed 4 h later. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed in sera. In addition, hepatomorphologic alterations, activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), gene expression of antioxidant enzymes and DNA integrity were evaluated in the liver. IAA administration did not show any alterations in any of the parameters available, except for a reduction of the gene expression for antioxidant enzymes by 55, 56, 27, and 28% for SOD, CAT, GPx, and GR upon T(500), respectively compared with the control. Several hepatic alterations were observed by DEN exposure. Moreover, IAA administration at 3 doses was shown to provide a total prevention of the active reduction of CAT and GR induced by DEN exposure compared with the control. IAA at T(500) was shown to give partial protection (87, 71, 57, and 90% for respectively SOD, CAT, GPx, and GR) on the down-regulation of the enzymes induced by DEN and this auxin showed a partial protection (50%) on DEN-induced DNA fragmentation for both parameters when compared to DEN alone. This work showed IAA hepatocarcinogenesis protection for the first time by means of a DEN-protective effect on CAT and GR activity, and by affecting antioxidant gene expression and DNA fragmentation.

Effect of indole-3-acetic acid administration by gavage and by subcutaneous injection on rat leukocytes.

This study was done to investigate the effect of indole-3-acetic acid (IAA) administered subcutaneously and by gavage on neutrophil function and cytotoxicity in neutrophils and lymphocytes. A gavage administration resulted in an increase in phagocytic capacity in neutrophils in a dose depended manner for 1 mg, 2 mg, 18 mg, and 40 mg of IAA per kg of body mass, respectively, compared with the control. Similarly, subcutaneous administration of IAA at 2, 18, and 40 mg per kg of body mass promoted a significant rise in phagocytosis by neutrophils. H2O2 production in neutrophils from treated rats by gavage was similar to those receiving subcutaneous IAA treatment, and did not show a significant difference between treatments and control. IAA treatment, whether by gavage or subcutaneous, did not produce an alteration in antioxidant enzyme activities or in glucose-6-phosphate dehydrogenase activity of either neutrophils or lymph nodes. Subcutaneous IAA administration did not alter the neutrophil and lymphocyte death as deduced by unaltered membrane integrity, DNA fragmentation and mitochondrial transmembrane potential, compared with controls. In conclusion, IAA administration either subcutaneously or by gavage could increase the phagocytic capacity by neutrophils and this acid administration did not have prooxidant effects or cytotoxic effects on neutrophils and lymphocytes.

Effect of Prototheca zopfii on neutrophil function from bovine milk.

This study was carried to investigate neutrophil function in the presence of Prototheca zopfii. For this purpose, bovine milk neutrophils were incubated in the absence (control) of and presence of P. zopfii, and then they were examined hydrogen peroxide (H(2)O(2)) production, antioxidant enzyme activities, and phagocytic capacity. Milk was collected from negative "California Mastitis Test" (CMT) quarter from three lactating Holstein cows after induction of leukocytosis with an intramammary infusion of oyster glycogen. H(2)O(2) production was measured using the phenol red method. Catalase activity was measured following H(2)O(2) reduction at 240 nm and the activity of glutathione reductase was determined by measuring the rate of NADPH oxidation at 340 nm. P. zopfii death was assessed by fluorescent microscopy using acridine orange assay and by colony forming units (CFUs). Comparisons between the groups were initially performed by analysis of variance (ANOVA). Significant differences were then compared using Tukey's test with a significance coefficient of 0.05. Hydrogen peroxide production, catalase and glutathione reductase activities by neutrophils incubated in presence of P. zopfii were stimulated five times, 21% and 27% respectively, compared to the unstimulated-neutrophils. Neutrophils did not affect P. zopfii death as shown by microscopy and CFUs. These observations led to the conclusion that the P. zopfii promote a high increase of H(2)O(2) production by neutrophils from bovine milk during algae exposition accompanied by increase of antioxidant enzyme activities; however, this process did not affect P. zopfii death.

Effect of indole acetic acid administration on the neutrophil functions and oxidative stress from neutrophil, mesenteric lymph node and liver.

This study was done to investigate the effect of the in vivo administration of indole acetic acid (IAA) on the neutrophil function, the activities of antioxidants enzymes in neutrophils, the mesenteric lymph node and on the oxidative stress in liver and plasma. The animals received subcutaneous administration of IAA in a phosphate-buffered saline (the control group received only the phosphate-buffered saline). The other groups received IAA at concentrations of 1 mg (T1), 2 mg (T2) and 18 mg (T3) per kg of body mass per day. Administration of IAA in both treatments T2 and T3 promoted a significant rise in the phagocytic capacity of neutrophils (by 51%), in comparison with the control. Another alteration was observed in antioxidant enzyme activities of the neutrophil and lymph node. But in the liver, the treatments imposed a significant decrease in the activity of catalase of 19% and 30% for T2 and T3, respectively, in comparison with the control. A similar effect was observed in the activity of hepatic glutathione peroxidase for T3 where a significant decrease of 31%, compared with the control, was obtained. The IAA did not show another significant alteration of the activities of superoxide dismutase and glutathione reductase activities in liver. The hepatic lipid peroxidation level, available by reactive products with thiobarbituric acid, has shown a significant decrease of 27% and 29% with T1 and T3 respectively, in comparison with the control. IAA treatment did not show a significant alteration in reduced glutathione contents in comparison with the liver and plasma controls. In conclusion, the IAA administration has a good potential animal utilization for increasing the phagocytic capacity with no prooxidant effect.

Indole-3-acetic acid increases glutamine utilization by high peroxidase activity-presenting leukocytes.

Indole-3-acetic acid (IAA) is toxic for human tumor cells and in association with horseradish peroxidase (HRP) can be used as a new prodrug/enzyme combination for targeted cancer therapy. The toxic effect of IAA on neutrophils, macrophages and lymphocytes is associated with cell peroxidase activity, which is high in neutrophils and low in lymphocytes. The effect of IAA on glucose and glutamine metabolism in leukocytes presenting different peroxidase activities: neutrophils, thioglycollate-elicited macrophages and lymphocytes was investigated. A time-course effect (from 6 to 48 h in culture) of IAA on glucose and glutamine metabolism of neutrophils, thioglycollate-elicited macrophages, and lymphocytes was then carried out. Addition of IAA (0.25 mM) did not have a marked effect on glucose utilization and lactate formation by the three cell types but it raised glutamine consumption and glutamate production by neutrophils and macrophages. IAA had no effect on glutamine consumption and glutamate production by lymphocytes. A strong relationship was found between glutamine utilization (0.999) and glutamate production (0.999) and peroxidase activity. IAA did not change the activities of hexokinase, glucose-6-phosphate dehydrogenase, citrate synthase, lactate dehydrogenase, and phosphate-dependent glutaminase of 24 h cultured neutrophils and lymphocytes. The effect of IAA (1 mM) on glucose and glutamine metabolism was also investigated by 1 h incubated leukocytes in PBS. IAA did not affect glucose and glutamine metabolism of lymphocytes but enhanced glucose and glutamine metabolism by 1 h incubated neutrophils and thioglycollate-elicited macrophages. IAA caused a marked increase on oxygen consumption by neutrophils, which was more pronounced in the presence of the glutamine as compared to glucose. The stimulation of oxygen consumption leads to a reduction in NADH/NAD+ ratio that activates the flux of substrates through the Krebs cycle. Since glutamine is mainly metabolized through the left hand side of the Krebs cycle, a reduction in the redox state of the cells may accelerate the flux of substrates through glutaminolysis. The toxic results presented here show that the affect of IAA in association with peroxidase involves activation of glutamine metabolism.