• Food Science of Animal Resources
• /
• v.39 no.1
• /
• pp.13-22
• /
• 2019

This study aimed to develop a bile-responsive expression system for lactobacilli. The promoters of four genes, encoding phosphoenolpyruvate-dependent sugar phosphotransferase (mannose-specific), L-lactate dehydrogenase (LDH), HPr kinase, and D-alanine-D-alanine ligase, respectively, which were highly expressed by bile addition in Lactobacillus johnsonii PF01, were chosen. Each promoter was amplified by polymerase chain reaction and fused upstream of the ${\beta}$-glucuronidase gene as a reporter, respectively. Then, these constructs were cloned into E. coli-Lactobacillus shuttle vector pULP2, which was generated by the fusion of pUC19 with the L. plantarum plasmid pLP27. Finally, the constructed vectors were introduced into L. plantarum for a promoter activity assay. The LDH promoter showed the highest activity and its activity increased 1.8-fold by bile addition. The constructed vector maintained in L. plantarum until 80 generations without selection pressure. A bile-responsive expression vector, $pULP3-P_{LDH}$, for Lactobacillus spp. can be an effective tool for the bile-inducible expression of bioactive proteins in intestine after intake in the form of fermented dairy foods.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.6
• /
• pp.840-846
• /
• 2021

To improve the bile salt and acid tolerance of probiotics against gastrointestinal stresses, we investigated the effects of soybean lecithin and whey protein concentrate (WPC) 80 on the bile salt tolerance of Lacticaseibacillus paracasei L9 using a single-factor methodology, which was optimized using response surface methodology (RSM). The survival rate of L. paracasei L9 treated with 0.3% (w/v) bile salt for 2.5 h, and combined with soybean lecithin or WPC 80, was lower than 1%. After optimization, the survival rate of L. paracasei L9 incubated in 0.3% bile salt for 2.5 h reached 52.5% at a ratio of 0.74% soybean lecithin and 2.54% WPC 80. Moreover, this optimized method improved the survival rate of L. paracasei L9 in low pH condition and can be applied to other lactic acid bacteria (LAB) strains. Conclusively, the combination of soybean lecithin and WPC 80 significantly improved the bile salt and acid tolerance of LAB. Our study provides a novel approach for enhancing the gastrointestinal tolerance of LAB by combining food-derived components that have different properties.

• Journal of radiological science and technology
• /
• v.31 no.2
• /
• pp.149-159
• /
• 2008

The purposes of this study are to analyze abnormal dilatation of the extrahepatic bile ducts by using transabdominal ultrasound, to confirm the existence of bile ducts diseases and their interrelationship, and for it to give a new theoretical basis for the technical access to extrahepatic bile ducts, upon which to analyze the ripple effects of the scan training. After teaching technical access process based on the new theory about extrahepatic bile duct to the thirty students who are studying ultrasonography, we allocated three hours per one student (30 mins ${\times}$ 6 times) to focus on the training of scanning skill. Training has been performed by one-to-one method. For evaluation, all the students have to perform the scans on (1) confluence of the right and left hepatic ducts (extrahepatic bile ducts and cystic duct), (2) the suprapancreatic bile duct, (3) the intrapancreatic bile duct, (4) intrapapilla Duct, based on the clearly divided concept. The existing training and methods have had low confidency about transabdominal ultrasonography of the extrahepatic bile duct and had limitation with which they could image only the suprapancreatic bile duct. The evaluation after finishing the train based on the new theory, however, all the students (30students) can access to (1) confluence of the right and left hepatic ducts(extrahepatic bile ducts and cystic duct), (2) the suprapancreatic bile duct objectively. 24 students can access to (3) the intrapancreatic bile duct and only one student can even make an image for (4) the intrapapilla Duct Though the evaluation on extrahepatic bile duct has to be performed with multi-sided method considering intrahepatic cause, bile duct cause and pathophysiological cause, only if we can image the extrahepatic bile duct to ampular of Vater objectively and confidently, we can greatly reduce invasive procedure such as ERCP, which is for the purpose of simple differential diagnosis and painful to the patients. Therefore if we concentrate on the scanning train based on the new theory to raise the confidency about ultrasonography, the effect will be doubled.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.30 no.1
• /
• pp.31-38
• /
• 1997

Changes in bile acid binding rapacity in vitro and physical properties of alginate extracted from sea tangle (Laminaria spp.) and residue after extracting alginate (RAEA) were investigated. For the purpose, extraction conditions controlled under 1, 3 and $5\%$ of sodium carbonate solution, and 1, 3, 5 and 10 hours of extraction time at $60^{\circ}C$. The less sea tangle had particle size and the higher concentration of sodium carbonate solution increseded, the more yield of alginate gained. High concentration of sodium carbonate solution and long extraction time resulted in weakly binding capacity in vitro by alginate. Among four bile acids, binding capacites with alginate were in the order of cholic, taurocholic acid>glycocholic acid>deoxycholic acid. The binding capacity of RAEA was rated at almost same degree of alginate. For increasing the binding capacity of bile acids by alginate, it was subject to high viscosity and degree of polymerization.

• The Korean Journal of Nuclear Medicine
• /
• v.19 no.2
• /
• pp.25-41
• /
• 1985

간흡충증은 한국 및 동남아세아에 널리 분포되어 있는 중요한 풍토병의 하나로서 진단은 전통적으로 분변검사에 의존하여 오고 있다. 최근 들어 담관내 간흡충 및 이의 합병증의 진단을 위해 ERCP 및 contrast cholangiogram등이 시도되었으나 실제에 이용하기에는 많은 제약이 있다 하겠다. 저자는 간흡충증에 있어서 최근 소개된 $^{99m}Tc-DISIDA$ hepatobiliary scintigraphy의 진단적 의의를 규명하고자 1982년부터 1983년까지 고신의대 부속 복음병원에서 검진한 간흡충증 환자 90명을 대상으로 하여 $^{99m}Tc-DISIDA$ hepatobiliary scintigraphy와 formalin-ether 원침법에 의한 분변검사를 시행하였다. $^{99m}Tc-DISIDA$ scintigram소견은 간내 담관 bile flow 및 총수담관 bile flow defect, 그리고 간세포기능의 정도에 따라 자료를 분석하였고 그외 합병증의 진단은 병록 기록, 수술 소견, ERCP 등에 의존하여 결론을 얻었다. $^{99m}Tc-DISIDA$ scintigraphy는 특이한 양상의 bile flow dynamics를 나타내었으며 간내 주담관의 intermittent irregular focal bile flow defect 및 tile flow stasis를 나타내고 말초담관의 bile flow defect는 경미하고 60분 내에 담관 bile flow activity의 완전한 배설을 나타내는 경우를 mild pattern, 간내 담관의 심한 irregular bile flow dynamics 및 간내담관의 심한 irregularity (담관내벽의 심한 불규칙성), 총수담관, 간내 주담관 및 말초 담관까지 심하게 irregular bile flow stasis를 나타내며 bile flow activity의 완전 베설이 $60\sim90$분사이 혹은 90분이상까지 인지된 경우를 moderate-severe pattern으로 분류하였다. 1) 분변검사상 간흡충증은 95검사중 70검사(환자 86명중 67명)에서 양성을 보여 분변 충난검사의 양성율은 73.7%였고 음성율은 26.3%였다. 2) $^{99m}Tc-DISIDA$ hepatobiliary scintigraphy는 90명중 70명에서 특이한 Cs-bile flow양성을 보였으며 양성율은 77.8%였으며 음성율은 22.2%였다. 3) $^{99m}Tc-DISIDA$ hepatobiliary scintigram양성율을 나타낸 환자 70명중 11명은 mild pattern, 59명은 moderate-severe pattern을 나타냈으며 그중 21명은 여러가지 간세포 기능 및 담관에 영향을 미치는 질환과 합병했지만 특이한 Cs-bile flow pattern을 dominent하게 나타내었으며 합병된 여러 질환들도 bile flow pattern상 인지 할 수 있었다. 4) $^{99m}Tc-DISIDA$ hepatobiliary scintigram 음성율을 나타낸 환자 20명중 8명은 만성간염, 5명은 간경변증, 3명은 재발성 농양성담관염(recurrent pyogenic cholangitis)과 간내담도의 stricture 및 담관담석증이 합병되었으며 scintigram상 합병증의 pattern을 나타냈고 4명에서는 low CBD obstruction을 나타내었으며 후에 CBD stone, CBD carcinoma, gall bladder Ca.의 porta hepatis 전이 및 clonorchis worms의 cluster에 의한 obstruction이 operation 및 ERCP로서 진단 되었다. 5) $^{99m}Tc-DISIDA$ hepatobiliary scintigraphy pattern은 현재의 자각증상과 관계된 dominent disease를 나타내었으며, 공간 점유병소도 multiple project images를 시행하므로서 쉽게 발견할 수 있었다. 이와 같이 간흡충증에 있어서 $^{99m}Tc-DISIDA$ hepatobiliary scintigram은 환자의 자각증상과 관계된 질환을 규명하는 데 필요한 정보를 얻었을 수 있었으며 간내담관의 damage정도를 규명하는데 필요한 procedure임이 판명되었다.

• Applied Microscopy
• /
• v.29 no.4
• /
• pp.437-450
• /
• 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.

• YAKHAK HOEJI
• /
• v.42 no.6
• /
• pp.572-575
• /
• 1998

Chitosan has been known to have hypocholesterolemic and hypolipidemic effects in animal studies. Chitosan also absorbs bile acids in vitro and in vivo, which might result in the hypocholesterolemic action. Trialkyl chitosan derivatives were prepared and tested for bile acid absorption activity in vitro. The derivatives showed enhanced absorption capacities which were comparable to cholestyramine.

• Journal of the Korean Chemical Society
• /
• v.42 no.3
• /
• pp.266-276
• /
• 1998

Cholesterol supersaturation in bile, which causes gallstone formation, is the result of low bile acid secretion or high cholesterol secretion. The quantitative analysis of cholesterol, bile acids and sterols which are precursors of cholesterol have been used to examine the changes in bile component. We described a simple, sensitive and reproducible method for simultaneous determination of cholesterol, five bile acids and seven sterols in human bile juices and gallstones by capillary column gas chromatography-mass spectrometry (GC/MS). Clinical samples were hydrolyzed by alcoholic KOH, extracted twice (pH 14 and 1) and derivatized to trimethylsilyl (TMS) ether with $MSTFA/NH_4I$ (N-methyl-N-trimethylsilyltrifluoroacetamide/ammonium iodide) mixture in order to be detected on the GC/MS. The good quality control data were obtained through within-a-day and day-to-day test (RSD values were 1.72-13.79, 0.68-14.10, respectively) and the recovery range of them was 73.56-96. 95 Using this method, biliary and gallstone compositions in the patients with intrahepatic stones were analyzed. The amounts and its relative distribution of cholesterol, bile acids and sterols showed different pattern in bile juices and gallstones.

• Korean Journal of Microbiology
• /
• v.47 no.3
• /
• pp.231-240
• /
• 2011

The objective of this study was to evaluate the acid and bile tolerance, bile salt hydrolase (BSH) activity, and cholesterol assimilation ability of lactic acid bacteria isolated from mustard leaf kimchi. MLK11, MLK22, MLK27, MLK41, and MLK67 were relatively acid- and bile-tolerant strains, with more than $10^5$ CFU/ml after incubation in simulated gastric juice and intestinal fluid, while MLK53 was the most sensitive strain to acid and bile. Strains MLK22 and MLK67 deconjugated the highest level of sodium glycocholate with more than 3.5 mM of cholic acid released, while deconjugation was lowest by strains MLK13 and MLK41 which released only 1.35 mM and 1.16 mM, respectively. Specially, strains MLK22 and MLK67 showed higher deconjugation of sodium glycocholate compared to sodium taurocholate and conjugated bile mixture. Although strains MLK22 and MLK67 exhibited maximal BSH activity at the stationary phase, MLK22 had somewhat higher total BSH activity compared to MLK67 towards both sodium glycocholate and sodium taurocholate. Meanwhile, cholesterol removal varied among tested strains (p<0.05) and ranged from 5.22 to 39.16 ${\mu}g$/ml. Especially, MLK67 strain assimilated the highest level of cholesterol in media supplemented with 0.3% oxgall, cholic acid, and taurocholic acid (p<0.05). According to physiological and biological characteristics, pattern of carbohydrate fermentation, and 16S rDNA sequence, strain MLK67 that may be considered as probiotic strain due to acid and bile tolerance and cholesterol-lowering effects was identified as Pediococcus pentosaceus MLK67.

• Korean Journal of Environmental Agriculture
• /
• v.18 no.3
• /
• pp.221-228
• /
• 1999

This study was carried out to extract the natural antioxidants from Bovine bile and to investigate their effects on various antioxidant activities. It also characterized the patterns of antioxidants by GC/FID and GC/MS. The antioxidative activities and chemical structure of the antioxidant were elucidated by examining the effects of biological activity and the analysis of GC/MS. The antioxidant materials extracted from bovine bile were isolated and purified by silica gel column chromatography and TLC. It was confirmed that there were effects of antioxidants such as Xanthine Oxidase(XO) and Glutathione-S-Transferase(GSH-T) on antioxidative activities. When they were compared with BHT, bile extracts showed the relative effects of 51.2% on the antioxidant activity, the inhibition effects of 48.3% on XO activity, and the synergism effects of 85.7% on the GSH-T activity. According to the results of investigation at neuron cell of mouse, the rate of cell activity in the treatment of 6mM glutathione was 96%, While it in the treatment of 140mg of bile extract was 78%. Based on the TLC analysis of EtOAc extracts from the Bovine bile, the antioxidant activity appeared at $R_f$ value, 0.72. These results suggested that the antioxidant may be coprostan 3-ol.