Influence of reduction of heme a and Cu(A) on the oxidized catalytic center of cytochrome c oxidase: insight from organic solvents.

Purified bovine heart cytochrome c oxidase (CcO) has been extracted from aqueous solution into hexane in the presence of phospholipids and calcium ions. In extracts, CcO is in the so-called "slow" form and probably situated in reverse micelles. At low water:phospholipid molar ratios, electron transfer from reduced heme a and Cu(A) to the catalytic center is inhibited and both heme a3 and Cu(B) remain in the oxidized state. The rate of binding of cyanide to heme a3 in this oxidized catalytic center is, however, dependent on the redox state of heme a and Cu(A). When heme a and Cu(A) are reduced, the rate is increased 20-fold compared to the rate when these two centers are oxidized. The enhanced rate of binding of cyanide to heme a3 is explained by the destabilization of an intrinsic ligand, located at the catalytic site, that is triggered by the reduction of heme a and Cu(A).

The influence of omega-3 polyunsaturated fatty acids (omega-3 pufa) on lactobacilli adhesion to the intestinal mucosa and on immunity in gnotobiotic piglets.

The effect of application of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on intestinal colonization by Lactobacillus paracasei and on cellular immunity has been investigated in gnotobiotic pigs. The administration of polyunsaturated fatty acids positively affected the adhesion of Lactobacillus paracasei to the jejunal mucosa of gnotobiotic piglets. When compared to the control group, the number of Lactobacillus paracasei adhering to the jejunal mucosa was by 12% higher in piglets of the experimental group (5.10 log 10/cm2 vs. 4.55 log 10/cm2). The respective counts of Lactobacillus paracasei adhering to the ileal and colonic mucosa of 28 day old gnotobiotic piglets reached 4.45 and 5.05 log 10/cm2 in group C and 4.44 and 4.95 log 10/cm2 in group E. Omega-3 PUFA supplementation increased the phagocytic activity of neutrophils by almost 100% on day 28 of life as well as the subpopulations of lymphocytes (CD8) in the peripheral blood of germ-free piglets on day 21 of life. Our results indicate that the action of probiotics in the gut may be modulated by dietary PUFA. The stimulatory effect of PUFA upon adhesion of lactobacilli could be used for enhancing the effectiveness of probiotics in inhibiting digestive tract pathogens.

Effect of silicate minerals (zeolite, bentonite, kaolin, granite) on in vitro fermentation of amorphous cellulose, meadow hay, wheat straw and barley.

The objective of the present experiment was to determine the effects of addition of silicate minerals, zeolite (Z), bentonite (B), kaolin (K), granite (G) on the rumen fermentation parameters, total gas, methane, total and individual volatile fatty acids (VFA) and hydrogen recovery in rumen fluid inoculum from sheep. Different materials (0.25 g) meadow hay (MH), wheat straw (WS), barley (BA) and amorphous cellulose (AC) were used as substrates. Silicate minerals (0.1 g) were added to the fermentation bottles containing substrates and rumen fluid inoculum and incubated for 72 h in vitro. The gas production technique simulates fermentation in the rumen was used to determine fermentation parameters. The total gas production was significantly higher compared to control for MH plus B (MHB), MH plus G (MHG), WS plus Z (WSZ), WS plus B (WSB), WS plus K (WSK), WS plus G (WSG), AC plus B (ACB), AC plus G (ACG), BA plus Z (BAZ), BA plus B (BAB), BA plus K (BAK), BA plus granite (BAG). Significant differences of the methane production were found between the controls, WSG, BAB and BAK. The total VFA concentration was increased in ACG (83.1 mM). The acetate: propionate (A:P) ratio of the control and additives ranged between 3.1 and 3.6 for MH, 2.7 and 3.5 for WS, 1.6 and 1.8 for AC and 2.3 and 2.9 for BA. It was concluded that the silicate minerals had no appreciable effect on the methane production, however, they support the microbial metabolism by influencing (bentonite, granite) and slightly influencing (zeolite, kaolin) the rumen fermentation.

Effect of dolomite, magnesium oxide (MgO) and chalk (CaCO3) on in vitro fermentation of amorphous and crystalline cellulose and meadow hay using inoculum from sheep.

Some minerals can influence some biochemical parameters of rumen fermentation. The objective of this experiment was to determine the effect of different amounts (0.1, 0.25 and 0.5 g) of dolomite and to compare the effect of dolomite, magnesium oxide (MgO) and chalk (CaCO3) upon the end products of rumen fermentation in vitro. Amorphous and crystalline cellulose as well as meadow hay were used as substrates and incubated with buffered rumen fluid in sealed fermentation bottles. In dependence on the amount of dolomite and the kind of substrate an inhibitory effect of dolomite on methane production was evident. Significant differences of methane production were found between the controls, crystalline cellulose and meadow hay with 0.5 g of dolomite. An increase of total gas production was observed for cellulose with both 0.25 and 0.5 g of dolomite and also for meadow hay with 0.5 g of dolomite. It can be concluded that there was a remarkable effect of dolomite on methane production and also a slight effect of magnesium oxide and chalk as compared to the effect of dolomite on the fermentation parameters of incubated substrates.