• BMB Reports
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• v.46 no.1
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• pp.47-52
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• 2013

Intestinal epithelial cell (IEC) apoptosis induced by hypoxia compromise intestinal epithelium barrier function. Both Akt and Hsp90 have cytoprotective function. However, the specific role of Akt and $Hsp90{\beta}$ in IEC apoptosis induced by hypoxia has not been explored. We confirmed that hypoxia-induced apoptosis was reduced by $Hsp90{\beta}$ overexpression but enhanced by decreasing $Hsp90{\beta}$ expression. $Hsp90{\beta}$ overexpression enhanced BAD phosphorylation and thus reduced mitochondrial release of cytochrome C. Reducing $Hsp90{\beta}$ expression had opposite effects. The protective effect of $Hsp90{\beta}$ against apoptosis was negated by LY294002, an Akt inhibitor. Further study showed that Akt phosphorylation was enhanced by $Hsp90{\beta}$, which was not due to the activation of upstream PI3K and PDK1 but because of stabilization of pAkt via direct interaction between $Hsp90{\beta}$ and pAkt. These results demonstrate that $Hsp90{\beta}$ may play a significant role in protecting IECs from hypoxia-induced apoptosis via stabilizing pAkt to phosphorylate BAD and reduce cytochrome C release.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.576-582
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• 2002

Probiotics are live microbial feed supplements which beneficially affect the host animal by improving its intestinal microflora. To select the best Lactobacillus spp. as a chicken probiotic, probiotic characteristics of 10 selected Lactobacillus strains isolated from chicken cecum or obtained from KCTC were investigated. The strains were examined for resistance to pH 2.0 and 0.3% oxgall, and adhesion to cecal mucus and cecal epithelial cells. All strains grew in MRS containing 0.3% oxgall. However, Lb. plantarum AYM-10, Lb. fermentum YL-3, AYM-3, and Lb. paracasei YL-6 showed relatively high resistance to 0.3% oxgall. Lb. fermentum YL-3, YM-5, AYM-3, and Lb. paracasei YL-6 survived 4 hours of incubation at pH 2.0. Lb. fermentum YL-3, KCTC 3112, and Lb. plantarum AYL-5 were strongly adhesive to cecal mucus, while the rest showed moderate or low adhesion. Lb. plantarum AYM-10, AYL-1, and AYL-5 had good adhering properties to cecal epithelial cells (30.7$\pm$10.82, 40.2$\pm$20.90, and 14.5$\pm$4.22, respectively). Lb. fermentum YL-3, AYM-3, and KCTC 3547 showed Intermediate adhesion ability, and Lb. plantarum showed better adhesion ability to cecal epithelial cells than Lb. fermentum. Attached Lb. fermentum YL-3 to cecum after 60 min incubation was confirmed using CLSM. Lb. fermentum YL-3 attached to a matrix which was composed of a mucus layer adjacent to intracrypts and pericryptal region. Some Lb. fermentum YL-3 bound to mucosal epithelial cells. From these results, Lb. fermentum YL-3 was selected as a chicken probiotic. In vivo trials of chicks inoculated with Lb. fermentum YL-3 had decreased Salmonella population in cecal contents and livers (p<0.5).

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.733-742
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• 2014

The glucagon-like peptide 2 (GLP-2) that is expressed in intestine epithelial cells of mammals, is important for intestinal barrier function and regulation of tight junction (TJ) proteins. However, there is little known about the intracellular mechanisms of GLP-2 in the regulation of TJ proteins in piglets' intestinal epithelial cells. The purpose of this study is to test the hypothesis that GLP-2 regulates the expressions of TJ proteins in the mitogen-activated protein kinase (MAPK) signaling pathway in piglets' intestinal epithelial cells. The jejunal tissues were cultured in a Dulbecco's modified Eagle's medium/high glucose medium containing supplemental 0 to 100 nmol/L GLP-2. At 72 h after the treatment with the appropriate concentrations of GLP-2, the mRNA and protein expressions of zonula occludens-1 (ZO-1), occludin and claudin-1 were increased (p<0.05). U0126, an MAPK kinase inhibitor, prevented the mRNA and protein expressions of ZO-1, occludin, claudin-1 increase induced by GLP-2 (p<0.05). In conclusion, these results indicated that GLP-2 could improve the expression of TJ proteins in weaned pigs' jejunal epithelium, and the underlying mechanism may due to the MAPK signaling pathway.

• Korean Journal of Food Science and Technology
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• v.36 no.6
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• pp.991-994
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• 2004

Effects of formulation KTG075 from edible plants on intestinal function, particularly on Mucin 2 secretion, were examined by loperamide-induced constipation method using Sprague Dawley rats (SD rats, male). Crypt epithelial cells containing more mucus and mucus layer stained with alcian blue were significantly thicker in KTG075 group than control group. When Biogenex AM358 of antibody against Mucin 2 was used, crypt epithelial cells secreted more Mucin 2 in KTG075 group than control group. The Mucus layer at fecal surface was thinner and less mucus was recovered from mucosal surface in constipated rats than in KTG075 group. Mucus production of crypt epithelial cells and mucus contents at fecal and mucosal surfaces were reduced by loperamide-induced constipation. These results indicate formula KTG075 accelerates evacuation and activates intestines.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1773-1778
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• 2008

Two experiments were carried out to study the effects of corticosterone (CORT) administration on intestinal morphology and function of broilers. In both experiments, birds were randomly divided into two equal groups. One group was the control group (CTRL), and the birds were fed with a basal diet. The other was the experimental group (CORT), and the birds were fed with the basal diet plus 30 mg of CORT/kg diet. At 21 days of age, performance, morphological characteristics of intestine, D-xylose level in plasma, activities of digestive enzymes in digesta, digestibility of nutrients and 5-bromo-2-deoxyuridine (BrdUrd)-labeling index of intestinal epithelial cells were determined. CORT administration decreased feed intake, daily gain and feed conversion ratio (p<0.05). CORT also decreased duodenal and jejunal villus height (p<0.05) as well as crypt depth (p<0.05). The D-xylose level in plasma of CORT-treated broilers was lower than that of the control (p<0.05). CORT treatment caused a decrease in apparent digestibility of protein (p<0.05), whereas fat and starch apparent digestibilities were unaffected (p>0.05). CORT administration increased activities of trypsin and amylase (p<0.05), and decreased BrdUrd-labeling index of duodenal and jejunal epithelial cells (p<0.05). In conclusion, CORT administration impaired the normal morphology and absorptive capacity of the small intestine of broiler chickens.

• Korean Journal of Veterinary Service
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• v.21 no.4
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• pp.401-405
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• 1998

A male badger which showed depression and bloody diarrhea was submitted to Iksan Branch of Chonbuk Veterinary Service Laboratory for necropsy on May 1998. Grossly, paleness of the mucous membranes was observed and the small intestinal contents were hamorrhagic. The numerous tiny hookworms, sized in 5-10 mm and greyish-white in color, attached to the intestinal mucosa. Male bursa was well developed. Histologically, intestinal lumen was filled with hemorrhagic contents, which contained worms. The epithelial cells of the villi were underwent degeneration and lamina propria was infiltrated by lymphocytes and plasma cells, and goblet cells were hyperplastic. There were rounded cutting plates in the funnel-shaped buccal capsule and transverse striation on sheath in hookworm, ultrastructurally, which were consistent with Uncinaria sp. The shape of eggs were ellipsoidal and morula, and some eggs contained a mobile larva. It was concluded that this badger was infected with Uncinaria.

• Parasites, Hosts and Diseases
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• v.60 no.4
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• pp.241-246
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• 2022

Felids are the unique definitive host of Toxoplasma gondii. The intestine of felid is the only site for initiating Toxoplasma gondii sexual reproduction. T. gondii excretes millions of infectious oocysts from the intestine, which are the primary source of infection. There are many difficulties in developing vaccines and drugs to control oocyst excretion due to the lack of an appropriate experimental model. Here, we established an in vitro feline intestinal epithelial cell (IEC) infection system and an efficient animal model of T. gondii Chinese 1 genotype, Wh6 strain (TgCtwh6). The Kunming mice brain tissues containing TgCtwh6 cysts were harvested 42-day post-infection. The bradyzoites were co-cultured with cat IECs in vitro at a ratio of 1:10. Five 3-month-old domestic cats were orally inoculated with 600 cysts each. The oocysts were detected by daily observation of cat feces by microscopy and polymerase chain reaction. We found that the parasite adhered and invaded cat IECs in vitro, transformed into tachyzoites, and then divided to form rose-like structures. These parasites eventually destroyed host cells, escaped, and finished the asexual reproduction process. Schizonts associated with sexual reproduction have not been observed during development in vitro cultured cells. However, schizonts were detected in all infected cat intestinal epithelial cells, and oocysts were presented in all cat feces. Our study provides a feasible cell model and an efficient infection system for the following studies of T. gondii sexual reproduction, and also lays a foundation to develop drugs and vaccines for blocking excretion and transmission of oocysts.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.592-600
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• 2021

Probiotics can be processed into a powder, tablet, or capsule form for easy intake. They are exposed to frequent stresses not only during complex processing steps, but also in the human body after intake. For this reason, various coating agents that promote probiotic bacterial stability in the intestinal environment have been developed. Silk fibroin (SF) is a material used in a variety of fields from drug delivery systems to enzyme immobilization and has potential as a coating agent for probiotics. In this study, we investigated this potential by coating probiotic strains with 0.1% or 1% water-soluble calcium (WSC), 1% SF, and 10% trehalose. Under simulated gastrointestinal conditions, cell viability, cell surface hydrophobicity, and cell adhesion to intestinal epithelial cells were then measured. The survival ratio after freeze-drying was highest upon addition of 0.1% WSC. The probiotic bacteria coated with SF showed improved survival by more than 10.0% under simulated gastric conditions and 4.8% under simulated intestinal conditions. Moreover, the cell adhesion to intestinal epithelial cells was elevated by 1.0-36.0%. Our results indicate that SF has positive effects on enhancing the survival and adhesion capacity of bacterial strains under environmental stresses, thus demonstrating its potential as a suitable coating agent to stabilize probiotics throughout processing, packaging, storage and consumption.

• Journal of Veterinary Science
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• v.24 no.4
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• pp.53.1-53.12
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• 2023

Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied. Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection. Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity. Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner. Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.

• BMB Reports
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• v.45 no.8
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• pp.433-441
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• 2012

Human milk, which nourishes the early infants, is a source of bioactive components for the infant growth, development and commensal formulation as well. Human milk oligosaccharide is a group of complex and diverse glycans that is apparently not absorbed in human gastrointestinal tract. Although most mammalian milk contains oligosaccharides, oligosaccharides in human milk exhibit unique features in terms of their types, amounts, sizes, and functionalities. In addition to the prevention of infectious bacteria and the development of early immune system, human milk oligosaccharides are able to facilitate the healthy intestinal microbiota. Bifidobacterial intestinal microbiota appears to be established by the unilateral interaction between milk oligosaccharides, human intestinal activity and commensals. Digestibility, membrane transportation and catabolic activity by bacteria and intestinal epithelial cells, all of which are linked to the structural of human milk oligosaccharides, are crucial in determining intestinal microbiota.