Addition Of Sodium Taurocholate Research Articles

Influence of dietary sodium taurocholate on the growth performance and liver health of Nile tilapia (Oreochromis niloticus).

Bile acids (BAs) are a class of cholesterol-derived amphipathic molecules approved as new animal feed additives. However, the functional researches mainly focused on BAs mixture, and the influence of the individual BA on fishes was still limited. In the present study, Nile tilapia were fed basal diet with three levels of sodium taurocholate at 0mg/kg (CON), 300mg/kg (TCAL), and 600mg/kg (TCAH) for 8weeks. The results indicated that addition of sodium taurocholate did not significantly influence the growth performance. Instead, TCAH group had higher cholesterol accumulation with liver fibrosis. In TCAH group, the level of nuclear factor E2-related factor 2 (nrf2) signaling-associated oxidative stress factors significantly increased in the liver. Additionally, fish in TCAH group had the highest expression level of genes encoding endoplasmic reticulum (ER) stress and inflammatory cytokines in the liver. In conclusion, 300mg/kg of sodium taurocholate did not significantly influence the growth performance of fish, while 600mg/kg of sodium taurocholate markedly induced cholesterol accumulation and liver injury, suggesting that the application of taurocholic acid in aquafeed should be re-evaluated.

W/O/W multiple emulsions of insulin containing a protease inhibitor and an absorption enhancer: biological activity after oral administration to normal and diabetic rats

In this work, the biological effects of w/o/w multiple emulsions of medium-chain triglycerides containing sodium insulin alone or with a protease inhibitor, or with an absorption enhancer, or with a protease inhibitor and an absorption enhancer, were compared to a w/o/w multiple emulsion containing zinc insulin. The release mechanism of all multiple emulsions was the swelling–breakdown phenomenon after dilution of the emulsions under hypo-osmotic conditions. The biological effects after oral administration to normal and diabetics rats showed a larger decrease of glycemia with the multiple emulsions containing sodium insulin than with the multiple emulsion containing zinc insulin. However, there was no significant difference between the hypoglycemic effects induced by the emulsions containing sodium insulin. These results suggest that the aggregation state of insulin molecules might be the major factor responsible for increasing the extent of intestinal insulin absorption. Thus, the nature of the insulin plays a fundamental role and, at the concentration used in this work, the addition of sodium taurocholate was not able to modify its aggregation state in aqueous solution, as confirmed by circular dichroism studies.

Bile acid-induced depolarization of mitochondrial membrane potential preceding cell injury in cultured gastric mucosal cells.

Changes in energy metabolism elicited by sodium taurocholate and their relation to cell viability were determined in gastric mucosal cells. Cultured mucosal cells were labelled with rhodamine-123, a mitochondrial energization-sensitive fluorescence probe, or by propidium iodide, a fluorochrome which labels the nuclei of non-viable cells. The cells were observed under a fluorescence microscope with a laser scanning confocal imaging system. After the addition of sodium taurocholate at concentrations > 5 mol/L, mucosal cells showed a rapid and significant decrease in rhodamine-123 fluorescence. A decrease to 40% of the pretreated values at 30 min was seen with a concentration of sodium taurocholate of 7.5mmol/L. A marked increase in the percentage of propidium iodide-positive cells was noted when the concentration of sodium taurocholate exceeded 5mmol/L. However, the extent of the decrease in rhodamine-123 fluorescence was always greater than the increase in the percentage of propidium iodide-positive cells, suggesting that most of these gastric mucosal cells remained viable. It is therefore suggested that the decrease in rhodamine-123 fluorescence is largely due to the disturbed oxidative phosphorylation of mitochondria. Pretreatment of gastric mucosal cells with low concentrations of ethanol resulted in a significant cytoprotective effect against sodium taurocholate injury with significant prevention of a decrease in rhodamine-123 fluorescence. It is concluded that sodium taurocholate induces a depolarization of the mitochondrial membrane potential preceding cell injury and that the cytoprotective effect of ethanol relates to its attenuation of the uncoupling effect.

Hydrolysis of Pyridoxine-5'- -d-Glucoside by a Broad-Specificity -Glucosidase from Mammalian Tissues

Research was conducted to evaluate the ability of a broad-specificity beta-glucosidase in mammalian tissues to catalyze the hydrolytic release of free pyridoxine from pyridoxine-5'-beta-D-glucoside, a naturally occurring form of vitamin B6 in plant-derived foods. Activity was detected in liver and intestinal mucosa using tritiated pyridoxine glucoside as a substrate. In the rat and guinea pig, enzyme activity was greater in intestine than in liver or kidney while even greater activity was detected in human intestinal tissue. Reaction rates were, however, low in all tissues. Hydrolysis of the synthetic substrate 4-methylumbelliferyl-beta-D-glucoside was also greatest in intestinal tissue. The characteristics of the enzymatic hydrolysis of pyridoxine glucoside to pyridoxine included: (i) most activity in the soluble tissue fraction, (ii) a pH optimum of approximately 6.0, and (iii) inhibition caused by the addition of sodium taurocholate. These characteristics are very similar to those of the broad-specificity beta-glucosidase in mammalian tissues with respect to the hydrolysis of a variety of naturally occurring and synthetic substrates. The apparent Km was greater than 2 mM for pyridoxine glucoside hydrolysis by intestinal preparations of each species, which is much greater than expected intestinal concentrations derived from dietary sources. In vivo studies have indicated that the intestine is involved in the metabolic utilization of dietary pyridoxine glucoside. The results observed here suggest that an alternate process, possibly involving intestinal microorganisms, may also be involved in the in vivo hydrolysis of pyridoxine glucoside.

Recovery of spores of Clostridium difficile altered by heat or alkali.

The effect of heating or alkali-treatment on spore recovery in ordinary growth medium was examined for four strains of Clostridium difficile. Heating spores at 80 degrees C for 10 min produced 95.50-99.95% decreases in the recovery rates. Treatment with 0.1 N NaOH for 15 min produced 99.47 and 99.83% decreases in spore recovery rates for two of the four strains. The influence of either addition of lysozyme after treatment with sodium thioglycollate (thioglycollate-lysozyme method) or addition of sodium taurocholate (taurocholate method) on recovery of heat- or alkali-treated C. difficile spores was also examined. Viable spores of all strains altered by heating at 90 degrees C or 100 degrees C for 10 min could not be recovered at all by the taurocholate method. Nor did this method allow recovery of alkali-altered spores treated with greater than 0.2 N NaOH for 15 min. On the other hand, 10-47% of altered spores heated at 90 degrees C for 10 min were recovered by the thioglycollate-lysozyme method, and alkali-altered spores treated with 0.1-0.3 N NaOH for 15 min were as completely recovered by this method as untreated spores. These results indicate that the thioglycollate-lysozyme method is more effective than the taurocholate method for recovery of the heat- or alkali-altered C. difficile spores.

Laboratory diagnosis of Clostridium difficile-associated diarrhoea.

This paper reviews the various laboratory procedures available for the isolation and identification of Clostridium difficile and the detection of toxins produced by this organism. Laboratories should be selective in determining which patients require investigation for Clostridium difficile-associated diarrhoea. Transport and storage of stool specimens at 4 degrees C is recommended when delays in processing may occur. Tissue culture techniques are still the best method for detection of cytotoxin and a variety of cell lines can be used. Other methods for detecting cytotoxin, and methods for detecting other toxins are not sufficiently developed yet to warrant introduction into diagnostic laboratories. Culture techniques remain the most sensitive for diagnosis, particularly since the development of a variety of enrichment techniques. Cycloserine cefoxitin fructose agar is still adequate, although reduced concentrations of antimicrobial agents are necessary, and improvements, such as the addition of sodium taurocholate, increase the recovery of spores. Enrichment cultures have markedly increased isolation rates for Clostridium difficile but the significance of these isolates needs to be carefully evaluated. Until simpler and more reliable tests are available in clinical laboratories for the detection of toxins, the isolation of Clostridium difficile from patients with diarrhoeal disease should be considered paramount.

Effect of synthetic surfactants on drug absorption in the presence of a physiologic surfactant, sodium taurocholate, in rats.

The effect of synthetic surfactants and their binary mixture with a physiologic one, sodium taurocholate (STC), on the intestinal absorption of drugs was examined in rats by using the in situ recirculation technique. Synthetic surfactants investigated in this study were nonionic polysorbate 80 (PS-80), cationic cetyltrimethylammonium bromide (CTAB), and anionic sodium lauryl sulfate (SLS). The surfactant action on the mucosal membrane seemed to be the same magnitude as the stronger one of the two components, except in the case of CTAB, as estimated by the enhancement effect on sulfaguanidine absorption. The action of CTAB was increased in the presence of STC. The action of surfactants in the aqueous solution, an intraluminal action, was examined by using quinine as a model drug. The micellar interaction of the drug with an ionic surfactant, CTAB or SLS, was reduced by the addition of STC, whereas, that with a nonionic surfactant, PS-80, was increased as estimated by a dynamic dialysis technique. The inhibitory effect on quinine absorption was approximately proportional to the degree of the micellar complex formation. Two other model drugs, imipramine and sulfadimethoxine, were further examined and the possible mechanisms of these effects were discussed.

Experimental ulceration of rabbit antral mucosa

The mechanism of acute ulceration and the nature of mucosal protective mechanisms were investigated in rabbit antrum, which is more resistant to injury than fundus of the same species, with particular reference to the relationship between H+ back diffusion, tissue acidification, and occurrence of ulceration. Exposure of antral pouches to increasing luminal [H+] during short-term hemorrhagic shock caused progressive acidification of the mucosa as measured by a microelectrode in the lamina propria, but ulceration of the mucosa did not occur unless an unphysiologically high [H+] (225 mM) was used. Addition of exogenous pepsin to a luminal [H+] of 80 mM increased the rate of H+ back diffusion and the degree of mucosal acidification, but ulcerations did not develop, suggesting that endogenous pepsin does not account for the ulceration of fundic mucosa under the same experimental situation. In contrast, addition of sodium taurocholate (5 mM) to the same luminal [H+] (80 mM) produced ulcerations and increased the rate of H+ back diffusion. Systemic administration of acetazolamide, an inhibitor of carbonic anhydrase, enhanced the development of ulceration without increasing the rate of back diffusion or the degree of tissue acidification. The results suggest that rabbit antral mucosa is protected from excessive acidification and ulceration by its relative impermeability to H+. When the rate of H+ back diffusion is artificially augmented to a level encountered in the fundus in an ulcerogenic situation, profound acidification with ulceration also occurs in the antral mucosa. The data also suggest that carbonic anhydrase, an enzyme abundantly present in gastric mucosa, may have a protective function in the mucosa, possibly by contributing to the regulation of intracellular pH and/or to the maintenance of mucosal HCO3− secretion.

Diet and sterol biohydrogenation in the rat: Occurrence of epicoprostanol

The fecal sterols from rats fed several types of semipurified or commercial diets were analyzed by a combination of thin layer and gas liquid chromatography. In rats fed semipurified diets with lard, sucrose, and casein, increasing proportions of lard (0, 8, 20, 65%) enhanced the fecal coprostanol/coprostanol + cholesterol ratio (from 0.50 to 0.85). This ratio was reduced by replacing lard with triolein or a mixture of calcium oleate and linoleate (1:1) and did not change when trierucin was substituted. No coprostanol formation was observed in rats fed a diet with tripalmitin or tristearin. The addition of sodium hyodeoxycholate (0.5%) or cholestyramine (2%) to the basal diet was without effect on the coprostanol/coprostanol + cholesterol ratio in the feces. The addition of sodium taurocholate (0.2, 0.75, and 4%) strongly reduced coprostanol formation, while a chronic bile duct ligation led to an enhancement. Cholesterol feeding (0.05, 0.2, and 0.5% in the diet) slightly increased (from 51 to 66%) coprostanol formation. Trace amounts of epicoprostanol were generally found in the feces. However, in some cases a very high proportion (up to 60%) of this sterol was observed. Possible relationships between the presence of epicoprostanol and the nature of the diet are discussed.

Selenium deficiency in chicks induced by feeding yeast grown on methanol as a sole dietary protein source.

Chicks fed a diet containing the yeast grown on methanol as a sole dietary protein source suffered from exudative diathesis (ED) and pancreas abnormality. Activity of their plasma glutathione peroxidase (GSHpx) was lowered and fragility of their erythrocytes (FE) was increased, indicating that yeast did not contain sufficient amount of either selenium (Se) or vitamin E. Lesions in pancreatic tissues, i.e., necrosis of acinar areas, infiltration of eosino-philic granulocytes, and regeneration of acinar cells were recognized. Se contents in the yeast and in the diet containing the yeast were so low that they could not be detected by fluorescent method with sensitivity limit of 0.01 ppm. Addition of 0.5 ppm of Se to the diet was effective to prevent ED and pancreatic abnormality completely, to recover the activity of GSHpx, and to decrease FE, while addition of vitamin E was only effective to prevent ED and FE com-pletely, but not effective to recover the activity of GSHPS. Addition of sodium taurocholate was not effective to prevent ED, to decrease FE, or to improve fat digestibility, indicating that vitamin E absorption was not severely damaged in the chicks fed the yeast despite histo-pathological change in the pancreas.

Substrate specificity of human α- l-fucosidase

Human α- l-fucosidase is a soluble lysosomal enzyme which hydrolyzes α- l-fucose residues linked to the 2 position of galactose or the 3, 4, or 6 position of N-acetylglucosamine. Demonstration of activity towards natural oligosaccharide or glycosphingolipid substrates was achieved by measuring liberated l-fucose by coupling to fucose dehydrogenase and NAD and measuring NADH production spectrophotometrically. Activity of purified human spleen, brain, and cultured skin fibroblast or crude cell extracts towards 4-methylumbelliferyl-α- l-fucoside had a pH optimum of 4.5 to 5.5 and was unaffected by the presence of neutral detergents such as Triton X-100. However, the addition of sodium taurocholate or other bile salts to the incubation mixture caused a marked inhibition at pH 5 and a shift in pH optimum to the pH 6–7 region. Sodium taurocholate effected a threefold reduction in the apparent K m for α- l-fucosidase at pH 6.0, but studies on fucosidosis tissue (α-fucosidase deficiency) or subcellular fractions derived from rat liver failed to indicate the existence of a membrane-bound α- l-fucosidase. The response of other lysosomal hydrolases to the presence of bile salts was investigated and was found to be variable, perhaps depending upon the hydrophilic or hydrophobic nature of the natural substrate and/or the state of association of the active enzyme.

Metabolic Indices for Evaluating the In vitro Maintenance of Hymenolepis diminuta in the Presence and Absence of Various Additives

An in vitro maintenance system for H. diminuta was devised by modifying the cultivation procedure of Schiller (1965). In this diphasic maintenance system, tissue culture medium (Triple Eagle's or NCTC-135) was used, in lieu of whole blood, as the 30% supplement to the agar phase. This provides a more defined system suitable for studying the effects of various additives. Morphological criteria were established which aided in assessing the efficacy of media used for the maintenance of 6- and 8-day-old H. diminuta. When successfully maintained, worms exhibited an intact scolex and neck region, undulatory movements along the strobila as well as integumentary and strobilar integrity. A more sensitive method for evaluating the maintenance of 8-day-old worms employed metabolic indices. Wet weight, protein and glycogen levels for 8-day-old H. diminuta served as base-line data allowing estimation of protein and glycogen contents of each worm prior to maintenance. Following maintenance, ratio of final to initial protein and final to initial glycogen levels (metabolic indices). A metabolic index approaching or exceeding unity suggested a reasonably intact metabolism. The addition of sodium taurocholate to the maintenance media appeared beneficial to the worms by prolonging the retention of normal signs. A combination of additives, taurocholate-nucleosides-lipids, improved the maintenance of H. diminuta for periods exceeding 24 hr as determined by observational criteria and metabolic indices. However, addition of a lipid mixture, or a lipid mixture prepared with a low concentration of taurocholate was not beneficial over a 24 hr period. The maintenance system, observational criteria, base-line data and metabolic indices should be useful for future in vitro studies requiring long-term incubation.

Cerebroside galactosidase: a method for determination and a comparison with other lysosomal enzymes in developing rat brain.

Abstract— A method is described for assaying brain for cerebroside galactosidase activity. The enzyme was liberated by sonication and addition of sodium taurocholate, then by digestion with pancreatic enzymes. It was further purified by precipitation at pH 3. The enzyme was then incubated with an emulsion of galactose‐labelled cerebroside in taurocholate and oleate at pH 4·5, and the liberated galactose was determined by scintillation counting. The content of cerebroside galactosidase in rat brain at various ages has been determined. The enzyme was present before cerebroside appears in noticeable amounts (4 days) and the amount rose considerably during the period of active cerebroside deposition and myelination. The amount then remained at a high concentration even in the adult. Comparison with other lysosomal brain enzymes was made in the age study. Nitrophenyl galactoside hydrolase also increased during myelination but levelled off earlier; its activity paralleled the amount of ganglioside. Nitrophenyl glucoside hydrolase started at a lower level and decreased with age. Sulphatase activity rose during myelination, then decreased somewhat after 15 days. Ceramidase followed a pattern similar to that of nitrophenyl galactoside hydrolase; it is suggested that both of these enzymes reflect ganglioside metabolism. The relative amounts of brain enzymes in different states were determined as a function of age in the case of cerebrosidase, nitrophenyl galactoside hydrolase and sulphatase. The proportion found in the high speed supernatant fraction was low but increased after myelination. The proportion that could be ‘solubilized’ by sonication decreased after myelination but the values differed greatly for the three enzymes. This treatment solubilized one‐seventh of the cerebrosidase, half the nitrophenyl galactosidase and three‐quarters of the sulphatase.

Studies on fatty acid esters of pyridoxine. VI. Effects of surface-active agents on the gastro-intestinal absorption of pyridoxine 3,4-dipalmitate in man.

Pyridoxine 3, 4-dipalmitate (PIN-DP), insoluble in water, was little absorbed in man following oral administration of suspension in water or aqueous Tween 80. But, the absorption increased markedly with a solubilized solution with Tween 80. Previously, it was found that PIN-DP failed to permeate the intestinal tract unlike pyridoxine (PIN) which was the hydrolyzed product of ester in the tract. Hydrolysis of PIN-DP solubilized with Tween 80 increased ten times with the addition of sodium taurocholate or glycocholate. But, no increase was observed with the addition of sodium lauryl sulfate or bis (2-ethylhexyl) sulfosuccinate. PIN permeated through the everted sac of rat intestine following the addition of sodium taurocholate and lipase to a solubilized Tween 80 solution of PIN-DP.

Experience with Lactose-Bile Medium for the Detection of B. Coli in Water

The discovery of B. coli in 1885 by Escherich, coupled with the knowledge that it is a normal inhabitant of the intestine of man and many animals, and therefore is found in sewage and polluted water, has induced water-bacteriologists to pay more attention to its isolation than to any other part of their work. Various methods have been proposed and tried for the detection of this organism in water. The one considered most reliable and meeting with general approval has involved the planting of a portion of the sample, either directly or after revivifying in broth, into plates of lactose-litmus agar; and then putting those colonies developing B. coli characteristics through the different tests as recommended for the identification of this organism. This procedure although accurate, is at the best a slow, tedious, and laborious process. Moreover, it is impracticable when used by the supervisor of a public water-supply, whose duty it is to report on the daily sanitary condition of the water. For example, if two days are required for the water to reach the inhabitants of the city, and it takes seven days to learn the results by this process, the consumer will have used the water five days before he is informed of the danger. To obviate these difficulties, various rapid methods have been proposed and used by different water-analysists. Nearly all are based on the fact that B. coli, when grown in a fermentable sugar, generates a certain amount of gas of which a definite proportion is carbon dioxid. In many cases certain chemicals whose function it is to inhibit the growth of the water bacteria have been added to the sugar solution. The most common is phenol, either alone or in combination with hydrochloric acid. McConkey has recommended the addition of sodium taurocholate to agar; others have used this compound in Smith solution. It would seem that these different methods, all based on the same general principles, ought to give something like concordant results. But unfortunately this is not the case and they