• Title/Summary/Keyword: Bile culture

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Electron Microscopic Study on the Role of Actin Filaments during the Formation of Bile Canaliculi in Isolated Rat Hepatocyte Culture System (흰쥐에서 분리 배양한 간세포의 담세관 형성에 있어서 액틴미세섬유의 역할에 관한 전자현미경적 연구)

  • Park, Chang-Hyun;Chang, Byung-Joon;Uhm, Chang-Sub
    • Applied Microscopy
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    • v.29 no.4
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    • pp.437-450
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    • 1999
  • Bile canaliculi are the structure delivering bile secreted by hepatocytes into the bile passage. Bile secretion is mainly controlled by the cytoskeletal elements, mainly of actin in the microvilli, pericanalicular web. Most studies on the bile secretion have been done in viva situation, however, to control the various parameters in vitro culture system seem to be more useful. To set up an in vitro experimental system, the investigator isolated hepatocytes with an enzymatic method using a mixture of collagenase and hyaluronidase from normal Sprague-Dawley rat liver and cultured. Isolated hepatocytes were round and formed cords in culture. Microvilli covered the whole surface of hepatocytes. Bile canaliculi were formed between hepatocytes and were characterized by the presence of microvilli of various lengths and shapes mainly arising from small surface mounds. Actin filament core in the microvilli and pericanalicular actin web were incomplete. After cytochalasin D treatment, cultured hepatocytes were round but the surface were irregular with surfacen blebs, folds and grooves. Microvilli on the surface were scarce. Bile canaliculi were markedly dilated often with the detached junctional complexes. Bile canaliculi lacks microvilli almost completely and extended into the pericanalirular cytoplasm showing complex vacuolar and tubular structures by transmission electron mciroscopy. Pericanalicular actin web, intermediate filaments were hardly identified. Subsurface actin filaments were scattered scarcely under the cell membranes. These results suggest that hepatocytes isolated from rats can survive and form bile canaliculi in culture and the actin filaments are involved in the formation and/or maintenance of the bile canaliculi.

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Salmonella sp. 의 신속한 동정을 위한 증진배양의 개선에 관한 연구

  • Kim, Kee-Tae;Kim, Tae-Ue;Yook, Soon-Hak;Pek, Un-Hua
    • 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.

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Inhibitory Effects of Bile Acids on the Cholesterol Biosynthesis in Cultured Hepatocytes (배양 간세포내에서의 콜레스테롤 합성에 대한 담즙산의 저해효과)

  • Kim, Sung-Wan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.5
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    • pp.496-501
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    • 1992
  • The present work tested the inhibitory effects of bile acids on the cholesterol biosynthesis and the activity of HMG-CoA reductase in cultured rat hepatocytes. The uptake of bile acids in hepatocytes were increased in according to the different bile acid concentrations and culture times. The rate of cholesterol synthesis in cells were inversely decreased to the bile acid concentrations and culture times. As expected, insulin injection (4 units/100g body weight) showed an enhancing effect of the cholesterol synthesis and the HMG-CoA reductase activity. The addition of bile acids in medium of insulin-treated hepatocytes also showed the suppressing effect. This effect was directly confirmed in isolated hepatic icrosomes by the test of HMG-CoA reductase activity. In the test of $Na^+$,$K^+$-ATPase activity in the isolated hepatocyte membrane, only the cholic acid did not stimulate the enzyme system. The reason of such difference is not obvious, but this result indicates that the cholic acid could be absorbed by simple diffusion.

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Isolation and Characterization of Lactic Acid Bacteria from Kimchi, Korean Traditional Fermented Food to Apply into Fermented Dairy Products

  • Cho, Young-Hee;Hong, Sung-Moon;Kim, Cheol-Hyun
    • Food Science of Animal Resources
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    • v.33 no.1
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    • pp.75-82
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    • 2013
  • This study aimed to isolate lactic acid bacteria (LAB) from Kimchi and to identify suitable probiotic strain for application in fermented dairy product as a commercial starter culture. A total of 106 (LAB) strains were isolated from Kimchi collected from different regions in Korea and their phenotypic characteristics were assayed. Four isolates from MRS agar plates were selected and designated as DKL109, DKL119, DKL121 and DKL128. They were identified first by API 50 CHL kit and then 16S rRNA gene sequencing. DKL121 and DKL128 were identified as Lactobacillus paracasei and Lactobacillus casei, respectively. Other two isolates (DKL109 and DKL119) were identified as Lactobacillus plantarum. To estimate their applicability in dairy products, the characteristics including acid and bile tolerance, cold shock induced cryotolerance and enzymatic activities were determined. There was wide variation in ability of strains to acid tolerance, but no significant differences in bile tolerance, cold shock induced cryotolerance within selected strains. DKL119 and DKL121 showed the highest resistance to acid and bile and the highest ${\beta}$-galactosidase activity, respectively. When these two strains were used for yogurt preparation as a single starter culture, their viable cell counts reached to $1.0{\times}10^9CFU/mL$. Lactobacillus plantarum DKL119 showed faster acid development than commercial starter culture. Also storage trials at $10^{\circ}C$ showed that the viability of these strains was retained over 15 d. With these results, it was indicated that probiotics isolated from Kimchi can be used in yogurt manufacturing as a starter culture.

Effect of Corn Peptide on the Lipid Metabolism in Rats (옥수수 펩타이드가 흰쥐의 지질 대사에 미치는 영향)

  • Lee, Hai-Mi;Chang, Un-Jae
    • Journal of the Korean Society of Food Culture
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    • v.16 no.5
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    • pp.416-422
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    • 2001
  • This study was conducted to investigate the effects of corn peptide(CP) on lipid metabolism. Sprague-Dawley(S.D) male rats were assigned to three dietary groups {control diet(CD), high fat diet (HFD) & high fat com peptide diet(FCD)} and fed 4 weeks to examine the effects of CP. There were no significantly different in cholesterol concentrations in the liver among the groups. However, triglyceride(TG) concentrations of the FCD & CD significantly lower than the HFD. Fecal excretion of neutral steroids & bile acids of the FCD significantly higher than the CD & HFD. Serum total cholesterol TG & LDL-cholesterol concentrations of the FCD & CD significantly lower than the HFD. These results suggest the improvement of lipid composition in serum by CP might be inhibit of lipid absorption in intestine & increment of neutral steroids & bile acids excretion in feces.

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Comparison of Probiotic Characteristics in Lactobacillus acidophilus Strains

  • Oh, Se-Jong;Chai, Chang-Hun;Kim, Sae-Hun;Kim, Young-Jun;Kim, Hyung-S.
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.349-352
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    • 2004
  • Twelve strains of Lactobacillus acidophilus isolated from feces of human or animal sources were tested for probiotic properties such as cholesterol assimilation, bile and acid tolerances, and CLA production. Although the cultures showed some variation with respect to each test, the 12 strains could be classified into 3 groups based on their ability to assimilate cholesterol. The cholesterol assimilation showed positive correlation with bile tolerance and negative correlation with acid tolerance. The cholesterol assimilation of L. acidophilus strains may not be related to the deconjugation activity, but may in fact be attributed to its bile tolerance. CLA production by lactic acid bacteria (LAB) exhibited a wide variation that ranged from 2.69 to 7.64 mg/g fat. CLA production of Bifidobacterium longum ATCC 15707 was the highest among the LAB tested, but there was no evidence for differences in CLA production between genus and species.

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Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Comparison of Acid and Bile Tolerances, Cholesterol Assimilation, and CLA Production in Probiotic Lactobacillus acidophilus Strains

  • Oh, Se-Jong;Chai, Chang-Hun;Kim, Sae-Hun;Kim, Young-Jun;Kim, Hyung-S.;Worobo, Randy W.
    • Food Science of Animal Resources
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    • v.32 no.4
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    • pp.409-413
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    • 2012
  • This study aimed to compare the probiotic characteristics of twelve strains of Lactobacillus acidophilus including cholesterol assimilation and conjugated linoleic acid (CLA) production. Cholesterol assimilation exhibited some variation among L. acidophilus strains, which could be classified into three groups based on their assimilation levels (p<0.05). The high cholesterol assimilation group exhibited a significantly higher tolerance to 0.3 and 0.5% bile acid than the low cholesterol assimilation group (p<0.05). Cholesterol assimilation showed positive correlation with 0.5% bile tolerance, and a negative correlation with acid tolerance (p<0.01). Glycocholate deconjugation activity showed no relationship with cholesterol assimilation, whereas taurocholate deconjugation activity was shown to have negative correlation with cholesterol assimilation (p<0.05). CLA production by L. acidophilus strains exhibited a wide variation, ranging from 2.69 to 5.04 mg/g fat. CLA production of L. acidophilus GP1B was the highest among the tested strains, but there was no evidence for differences in CLA production in strain specificity. Based on these results, the cholesterol assimilation of L. acidophilus strains may not be related to deconjugation activity, but may in-fact be attributed to their bile-tolerance.

An Experimental Animal Model of Anomalous Pancreaticobiliary Duct Union (췌담관 합류이상의 실험동물 모델)

  • Han, Seok-Joo;Chang, Hang-Seok;Kim, Jong-Sung;Han, Jin-Soo;Kim, Ho-Geun;Hwang, Eui-Ho
    • Advances in pediatric surgery
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    • v.4 no.2
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    • pp.100-109
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    • 1998
  • The anomalous pancreaticobiliary duct union (APBDU) might cause the formation of choledochal cyst and malignancies of hepatopancreaticobiliary system. The purpose of this study is to make an experimental animal model of APBDU similar to that of human. One to two-month-old Mongrel dogs (n=12) were divided into two groups; the control group (n=2) had a sham operation performed, and in the experimental group (n=10) the end of distal ' common bile duct (CBD) was anastomosed to the side of the dorsal pancreatic duct making APBDD. Serum was obtained for chemical analysis on the 10th postoperative day. The dogs were sacrificed at the 5th week (n=3), the 6th week (n=3), the 7th week (n=2), the 8th week (n=2) and the 6th month (n=2) after the experimental surgery. With sacrifice, operative cholangiogram was taken, and bile juice was obtained for chemistry and bacterial culture. The en-bloc specimens of the hepatopancreaticobiliary system were removed for microscopic examination. Serum and bile juice amylase levels were elevated in the experimental group(n=10), but not in the control group(n=2). Operative cholangiograms of control group revealed no evidence of bile duct dilatation.. On the other hand, the bile duct in the experimental group was markedly dilated without any evidence of stenosis at the anastomosis site (n=10). Histologic examination of the hepatopancreaticobiliary system in the experimental group resembled the findings of choledochal cyst in human. The APBDU of this animal model can produce bile duct dilatation by pancreaticobiliary reflux. We think that this animal model can be potentially promising for the research about the APBDU associated hepatopancreaticobiliary diseases.

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Isolation and Identification of Lactic Bacteria Containing Superior Activity of the Bile Salts Deconjugation (담즙산 분해능이 뛰어난 젖산균의 분리 및 동정)

  • 하철규;조진국;채영규;허강칠
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.164-170
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    • 2004
  • The purpose of this study is to isolate probiotic lactic acid bacteria (LAB) that produced bile salts hydrolase. One hundred twenty strains were initially isolated from human feces. Based on their resistance of acid, tolerances of bile salts, and inhibitory activity against Escherichia coli, five strains were selected. A strain producing highest activity of bile salts hydrolase was identified as Lactoacillus plantarum using API carbohydrate fermentation pattern and 16S rRNA sequences, and named CK102. Lactobacillus plantarum CK102 survived at a level of 1.36${\times}$10$\^$8/ CFU/$m\ell$ in pH 2 buffer for 6 h and showed exhibited excellent bile tolerance. When L plantarum CK102 was cultured with E. coli in MRS broth, no viable cells of E. coli was detected after 18 h fermentation. These results suggest that Lactobacillus plantarum CK 102 may be commercially used for the probiotic culture.