• Journal of Dairy Science and Biotechnology
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• v.37 no.1
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• pp.40-48
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• 2019

Bifidobacteria are a prototype probiotic, which normally inhabit the intestinal tract of humans. In the present study, four species of Bifidobacterium isolated from the feces of infants were characterized. The tolerance for acid or bile salt, autoaggregation, and antibiotic resistance of the bacteria were examined. The four species were resistant to low pH, bile salts, and up to 3% bile acid. Autoaggregation rates were as high as 90%. The bacteria were consistently resistant to gentamicin, kanamycin, streptomycin, ciprofloxacin, and nalidixic acid. Due to their tolerance to environmental factors like acid and bile salts, B. longum MG723, B. breve MG729, B. bifidum MG731, and B. animalis subsp. lactis MG741 are potentially valuable as probiotics and may be useful for industrial application.

• Korean Journal of Poultry Science
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• v.27 no.1
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• pp.31-35
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• 2000

This study was investigated to observe Lactobacillus spp. population of poultry digestive organ, duodenum, ilium, cecum and colon by growing steps and to select of valuable lactobacilli as probiotics. Two strains of male broiler commercial chickens, Hybrid and Ross, were used to evaluate population of Lactobacillus spp. in intestinal tracts. Three strains of Lactobacillus were identified, and bile salts environment. The number of lactovacilli was the lowest in duodenum compared to other intestinal tracts which had similar population. Population of Lactobacillus was maintained constantly regardless growing steps after one week of age. Identification of Lactobacillus from cecum resulted in L.reuteri BC5, L. crispatus BC7, L.reuteri BC9. All strains was depressed in pH 1 and 2, although two strains could survive for one hour at pH 2. And they could survive at pH 4 for 4hours. In bile salts tolerance, L.reuteri BC5, L.crispatus BC7 were maintained for 2 hours, but the growth reduced from 2hours. Growth of L.reuteri BC9 was increased continuously. In conclusion, Lactobacillus of intestinal tracts were established at first week, and maintained constant population. They were influenced on severe acidic condition and bile salts. Cecal Lactobacillus has different growth charcteristics by strains.

• Toxicological Research
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• v.19 no.1
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• pp.67-72
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• 2003

Cholestatic liver injury results from the accumulation of toxic bile salts within the liver. The aim of the present study is to elucidate the changes in expression and cellular localization of apoptosis related proteins in the liver of bile duct-ligated (BDL) rat. Extrahepatic cholestasis was induced by double ligation of the common bile duct and cut between the ligatures. Animals were sacrificed at day 3 and at week 1, 2, 4, 6, and 8 after BDL. The number of TUNEL positive cells was increased significantly after 3 days of BDL, decreased over 2 weeks and remained constant thereafter. Fas expression was not changed and activation of caspase 8 did not occur. Fas immunoreactivity was exclusively observed in the cytoplasm of hepatocytes, indicating that Fas expressed in rat hepatocytes is a soluble form. Hepatocyte apoptosis was associated with Bax expression, which showed a peak at day 3 and decreased over time gradually. Immnunostaining of Bax was observed in hepatocytes and bile duct epithelial cells (BEC) of control and BDL rats. Bcl-2 was increased over time in BDL rats. These results suggest that apoptosis of hepatocytes in BDL rats is independent of Fas and controlled by Bax expression.

• Journal of Dairy Science and Biotechnology
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• v.29 no.1
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• pp.49-54
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• 2011

Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a $Ni^+$-NTA agarose column and characterized. The BSH enzyme of E. faecalis CU30-2 exhibited approximately 50 times higher activity against glycol-conjugated bile salts than tauro-conjugated bile salts having the highest activity against glycocholic acid. Considering the prevalence of E. faecalis strains in the human GI tract and glycol-conjugates dominated bile acid composition of human bile, further study is needed to investigate the impact of the BSH activity exerted by E. faecalis strains to the host as well as to the BSH producing strains.

• Journal of Pharmaceutical Investigation
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• v.24 no.2
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• pp.73-83
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• 1994

Applications of liposomes as a drug carrier for the oral delivery of poorly-absorbable macromolecular drugs have been limited, because of their instability in gastrointestinal environments including pH, bile salts, and digestive enzymes. Two polysaccharides, dextran(DX) and pullulan(PL), were introduced to the preformed liposomes in order to enhance the stability. Palmitoyl derivatives of polysaccharides, palmitoyldextran(PalDX) and palmitoylpullulan(PalPL), were synthesizd and introduced to the liposomes during preparation for the same purpose of stability. The effects of these polysaccharides coating were evaluated basically by physical properties of particle size distribution and optical microscopy, then compared with uncoated liposomes by the observations of both in vitro stability and in vovo absorption characteristics. The geometric mean diameters of polysaccharide-coated liposomes were greater than that of uncoated liposome, showing the outermost polysaccharide-coated layer under the optical microscopy. In vitro stabilities of uncoated or polysaccharides-coated liposomes were measured by turbidity changes in various pH buffer solutions containing sodium choleate as bile salts. While uncoated liposome was very sensitive to bile salts, polysaccharides-coated liposomes were stable in relatively higher concentrations of sodium choleate, giving the results of better stability of PalDX- and PalPL-coated liposomes than that of DX- and PL-coated liposomes. After liposomal encapsulation of acyclovir(ACV), an antiviral agent as a model drug, it has been administered orally to rats as dose of ACV 40 mg/kg. Plasma concentrations of ACV were assayed by HPLC and analyzed by model-independent pharmacokinetics. Pharmacokinetic parameters of Cmax, tmax, and [AUC] have been compared.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.133-138
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• 2001

In this study, an attempt was made to increase the survival rate of bifidobacteria entrapped in alginate in the gastrointestinal tract, and to investigate the potential industrial applications, for example lyophilized capsules and yogurt. First, the protective effect of various food additives on bifidobacterial survivability was determined after exposure to simulated gastric juices and bile salts. The additives used in this study were skim milk (SM), polydextrose (PD), soy fiber (SF), yeast extract (YE), chitosan (CS), $\kappa$-carageenan ($\kappa$-C) and whey, which were added at 0.6% concentration (w/v) to 3% alginate-bifidobacterial solution. In the simulated gastric juices and bile salts, the protective effect of 0.6% skim milk-3% alginate (SM-A) beads on the survival rate of bifidobacteria proved to be higher than the other additives. Second, the hydrogen ion permeation was detected through SM-A vessel without bifidobacterial cells at different SM concentrations (0.2%, 0.4%, 0.6%, 0.8%, and 1.0%). There were no differences in terms of the pH decrease in SM-A vessels at 0.6%, 0.8%, and 1.0% (w/v) SM concentrations. The survival rate of bifidobacteria in SM-A beads would appear to be related to the SM buffering capacity against hydrogen ions and its tendency to reduce the pore size of bead. In this experiment, the survival rate of bifidobacteria entrapped in beads containing 0.6% SM showed the highest viability after exposure to simulated gastric juices for 3h, thereby indicating that 0.6% SM is the optimum concentration fir 3% alginate bead preparation. Third, the effect of SM-A beads on the freeze-drying and yogurt storage for 10 days was investigated. SM-A beads were found to be more efficient for freeze drying and yogurt storage than untrapped cells and the alginate bead. Consequently, the survival rate of bifidobacteria entrapped in SM-A beads was increased in simulated gastric juices, bile salts and probiotic products, such as lyophilized capsules and yogurt, SM-A beads can be expected to produce high value probiotic products.

• Archives of Pharmacal Research
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• v.29 no.4
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• pp.323-327
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• 2006

The objective of this study was to examine the pharmacokinetics of organic cations in intrahepatic cholestatic rats. A pretreatment with $17{\alpha}$-ethynylestradiol was used to induce intrahepatic cholestasis, and tributylmethylammonium (TBuMA) was used as a representative model organic cation. When $[^3H]$TBuMA was intravenously administered, the AUC value for TBuMA was significantly increased by $79\%$ in cholestasis, and its total systemic clearance was consequently decreased by $46\%$. In addition, the in vivo hepatic uptake clearance of TBuMA from the plasma to the liver was decreased by $50\%$ in cholestasis. The concentration of bile salts in plasma was increased by 2.1 fold in cholestatic rats. Since TBuMA forms ion-pair complexes with anionic components such as bile salts, the decreased hepatic uptake of TBuMA in cholestasis may be due to a change in endogenous components, e.g., bile salts in the plasma. In isolated normal hepatocytes, the uptake clearance for TBuMA in the presence of cholestatic plasma was decreased by $20\%$ compared with normal plasma. Therefore, we conclude that the inhibition of the hepatic uptake process by the cholestasis may be in part due to the increased formation of ion-pair complexes of TBuMA with bile salts in the plasma.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.162-165
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• 2002

With the objective of identifying the commercial potential of new direct-fed microbials, several temperature-tolerant strains were isolated from cane molasses at $39^{\circ}C$ and tested for their tolerance to pH, bile salts, and a mixture of volatile fatty acids (acetic acid:propionic acid:butyric acid=6.5:2.0:1.5). It was found that the isolated strain DY 252 grew very well up to pH 2.0 and was resistant to relatively high concentrations of bile salts. Among the strains tested, DY 252 was least inhibited by the addition of volatile fatty acids to the growth medium at $39^{\circ}C$. Accordingly, it would appear that strain DY 252, identified as yeast Issatchenkia orientalis, may be a potential candidate for use as a microbial feed additive.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1505-1512
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• 2005

Bile salt deconjugation is the most biologically significant reaction among the bacterial alterations of bile acids in the gastrointestinal tract of human and animal. The responsible enzyme, bile salt hydrolase (BSH), catalyzes the hydrolysis of glycineand/or taurine-conjugated bile salts into amino acid residues and deconjugated bile acids. Herein we review current knowledge on the distribution of BSH activity among various microorganisms with respect to their biochemical and molecular characteristics. The proposed physiological impact of BSH activity on the host animal as well as on the BSH-producing bacterial cells is discussed. BSH activity of the probiotic strains is examined on the basis of BSH hypothesis, which was proposed to explain cholesterol-lowering effects of probiotics. Finally, the potential applications of BSH research are briefly discussed.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.647-651
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• 1996

The development of an enrichment method for the rapid and effective identification of Salmonella spp. in sewage or food was studied. As a growth factor for Salmonella, 10 mM cyclic adenosine monophosphate (cAMP) in trypticase soy broth with 0.6% yeast extract (TSBYE) increased cell number five-folds and 0.6% yeast extract in selenite broth increased cell number ten-folds of control. Bile salts in selenite broth was tested for the selection of S. enteritidis in a mixture with Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus plantarum and Escherichia coli. The latter four strains were effectively inhibited at 0.1% bile salt. A two-step culture method was used to enrich Salmonella spp.; a primary-enrichment and secondary- enrichment culture. At a primary-enrichment step, selenite broth with 0.6% yeast extract and 10 mM cAMP was used, and at a secondary-enrichment step, 0.1% bile salt was additionally used. Culture times of a primary- enrichment and a secondary-enrichment step were 8 hr and 6 hr, respectively. In this procedure, cell number increased from 10$^{0.3}$ to 10$^{8.5}$ with inhibition of other strains within 14 hr. In the case of an initial cell concentrarion as low as 10$^{-2}$ cfu/ml, a cell number increased to 10$^{7}$ cfu/ml by using a 10 hr primary-enrichment and 6 hr secondary-enrichment procedure.