• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1557-1564
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• 2009

Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.17-26
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• 2000

The regional distribution and relative frequencies of endocrine cells were studied immunohistochemically (PAP methods) in the alimentary tract and pancreas of the toad, Bufo bufo gargarizans Cantor using specific antisera against bovine Sp-1/chromogranin (BCG), serotonin, bombesin, gastrin, substance P (SP), somatostatin, insulin, glucagon, pancreatic polypeptide (PP), vasoactive intestinal polypeptide (VIP) and secretin. Nine kinds of endocrine cells were identified in this study. Spherical or spindleshaped immunoreactive (IR) cells were located in the gastric glands of stomach regions, in the basal portion of the epithelium of intestinal tract or esophagus, and in the exocrine or pancreatic islets with variable frequencies. In the alimentary tract, BCG-IR cells were found in the fundus and pylorus with rare and a few frequencies, respectively. Serotonin-IR cells were demonstrated in the whole alimentary tract including the esophagus. Bombesin- and SP-IR cells were restricted to the stomach regions and gastrin-IR cells were restricted to the pylorus. Somatostatin-IR cells were detected throughout the whole alimentary tract except for the large intestine, However, insulin-, glucagon-, PP-, VIP- and secretin-IR cells were not detected in the alimentary tract. In the pancreas of toad, the distribution and relative frequency of endocrine cells were similar to those of other mammals. Insulin-IR cells were located in the central portion of the pancreatic islets and interspaces of exocrine portions, and glucagon-, somatostatin- and PP-IR cells were detected in the marginal regions of the pancreatic islets and interspaces of exocrine. However, other IR cells were not found in the pancreas. In conclusion, the regional distribution and relative frequency of the endocrine cells in the alimentary tract and pancreas of the toad were similar to other anuran species but some differences which might be caused by feeding habits and species specification were also observed.

• Journal of fish pathology
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• v.25 no.3
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• pp.173-180
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• 2012

Vibriosis is one of the most prevalent fish disease belonging to the genus Vibrio. In present study, Vibrio sp. isolated from Japanese eel was confirmed as Vibrio scophthalmi using analysis of the genomic sequence of 16S rRNA. The major signs were hemorrhage of body surface and inner surface of abdomen, severe enteritis and retention of ascitis. Histopathological examination revealed blood cell degenerations in various organs (gills, liver, spleen, kidney, heart, intestine), exfoliate of intestinal epithelium, and congestion and hemorrhage in intestinal lamina propria. This is the first case report on V. scophthalmi infection in Japanese eel Anguilla japonica.

• Archives of Pharmacal Research
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• v.29 no.4
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• pp.318-322
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• 2006

Many quaternary ammonium salts are incompletely absorbed after their oral administration and may also be actively secreted into the intestine. However, the underlying mechanism(s) that control the transport of these cations across the intestinal epithelium is not well understood. In this study, the mechanism of absorption of quaternary ammonium salts was investigated using Caco-2 cell monolayers, a human colon carcinoma cell line. Tributylmethylammonium (TBuMA) was used as a model quaternary ammonium salts. When TBuMA was administrated at a dose of 13.3 imole/kg via iv and oral routes, the AUC values were $783.7{\pm}43.6\;and\;249.1{\pm}28.0{\mu}mole\;min/L$ for iv and oral administration, indicating a lower oral bioavailability of TBuMA $(35.6\%)$. The apparent permeability across Caco-2 monolayers from the basal to the apical side was 1.3 times (p<0.05) greater than that from the apical to the basal side, indicating a net secretion of TBuMA in the intestine. This secretion appeared to be responsible for the low oral bioavailability of the compound, probably mediated by p-gp (p-glycoprotein) located in the apical membrane. In addition, the uptake of TBuMA by the apical membrane showed a $Na^+$ dependency. Thus, TBuMA appears to absorbed via a $Na^+$ dependent carrier and is then secreted via p-gp related carriers.

• Parasites, Hosts and Diseases
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• v.27 no.2
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• pp.109-118
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• 1989

Production of circulating specific antibodies to the lung fluke (Paragenimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods, However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of p. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs(male and female), and eggs. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um thickness as the antigen and cat antisera (11~20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobensidine(DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1 : 500~1 : 2, 000 and 1 : 200~1 : 500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

• Journal of fish pathology
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• v.24 no.1
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• pp.11-18
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• 2011

This study was aimed to investigate the pathophysiological changes of olive flounder, Paralichthys olivaceus suffering from emaciation. A plasma osmolality was higher in the emaciated and control flounders than that of normal teleost, suggesting osmoregulatory failure in both of them. Also, the control in the same stock with emaciated flounder seem to be classified into a primary degree of emaciation. According to microscopic observations, the inflammatory responses were observed in the submucosal layer of anterior intestine, although the some of mucosal intestinal epithelium still remained. It was suggested that the pathological changes of the anterior part give rise to malabsorption of nutrients through the mucosa. In the posterior intestine and rectum, the mucosal epithelium were almostly sloughed off and severe inflammatory responses were observed in the submucosa. Immunoreaction for NKCC was not detected in the mucosal epithelial cells in intestine because of sloughing of epithelium. These changes would lead to functional disorder in the intestine, such as malabsorption of nutrients and osmoregulatory failure. Also important is to investigate the recovery phase.

• Proceedings of the Korean Nutrition Society Conference
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• 1995.11b
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• pp.11-34
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• 1995

Growth hormone (GH) plays a key role in regulating postnatal growth and can stimulate growth of animals by acting directly on specific receptors on the plasma membrane of tissues or indirectly through stimulating insulin-like growth factor (IGF)-I synthesis and secretion by the liver and other tissues. IGF-I and IGF-Ⅱ are polypeptides with structural similarity with proinsulin that stimulate cell proliferation by endocrine, paracrine and autocrine mechanisms. The initial event in the metabolic action of IGFs on target cells appears to be their binding to specific receptors on the plasma membrane. Current evidence indicates that the mitogenic actions of both IGFs are mediated primarily by binding to the type I IGF receptors, and that IGF action is also mediated by interactions with IGF-binding proteins (IGFBPs). Six distinct IGFBPs have been identified that are characterized by cell-specific interaction, transcriptional and post-translational regulation by many different effectors, and the ability to either potentiate or inhibit IGF actions. Nutritional deficiencies can have their devastating consequence during growth. Although IGF-I is the major mediator of GH's action on somatic growth, nutritional status of an organism is a critical regulator of IGF-I and IGFBPs. Various nutrient deficiencies result in decreased serum IGF-I levels and altered IGFBP levels, but the blood levels of GH are generally unchanged or elevated in malnutrition. Effects of protein, energy, vitamin C and D, and zinc on serum IGF and IGFBP levels and tissue mRNA levels were reviewed in the text. Multiple factors are involved in the regulation of intestinal epithelial cell growth and differentiation. Among these factors the nutritional status of individuals is the most important. The intestinal epithelium is an important site for mitogenic action of the IGFs in vivo, with exogenous IGF-I stimulating mucosal hyperplasia. Therefore, the IGF system appears to provide and important mechanism linking nutrition and the proliferation of intestinal epithelial cells. In order to study the detailed mechanisms by which intestinal mucosa is regulated, we have utilized IEC-6 cells, an intestinal epithelial cell line and Caco-2 cells, a human colon adenocarcinoma cell line. Like intestinal crypt cells analyzed in vivo or freshly isolated intestinal epithelial cells, IEC-6 cells and Caco-2 cells possess abundant quatities of both type Ⅰ and type Ⅱ IGF receptors. Exogenous IGFs stimulate, whereas addition of IGFBP-2 inhibits IEC-6 cell proliferation. To investigate whether endogenously secreted IGFBP-2 inhibit proliferation, IEC-6 cells were transfected with a full-length rat IGFBP-2 cDNA anti-sense expression construct. IEC-6 cells transfected with anti-sense IGFBP-2 protein in medium. These cells grew at a rate faster than the control cells indicating that endogenous IGFBP-2 inhibits proliferation of IEC-6 cells, probably by sequestering IGFs. IEC-6 cells express many characteristics of enterocyte, but do not undergo differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation after reaching confluency. We have demonstrated that Caco-2 cells produce IGF-Ⅱ, IGFBP-2, IGFBP-3, and an as yet unidentified 31,000 Mr IGFBP, and that both mRNA and peptide secretion of IGFBP-2 and IGFBP-3 increased, but IGFBP-4 mRNA and protein secretion decreased after the cells reached confluency. These changes occurred in parallel to and were coincident with differentiation of the cells, as measured by expression of sucrase-isomaltase. In addition, Caco-2 cell clones forced to overexpress IGFBP-4 by transfection with a rat IGFBP-4 cDNA construct exhibited a significantly slower growth rate under serum-free conditions and had increased expression of sucrase-isomaltase compared with vector control cells. These results indicate that IGFBP-4 inhibits proliferation and stimulates differentiation of Caco-2 cells, probably by inhibiting the mitogenic actions of IGFs.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1178-1182
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• 2008

The present study was designed to assess the influence of caeca on the availability of calcium, phosphorus, magnesium, manganese and copper from soybean, sunflower, rapeseed, sesame, fish and meat cum bone meal in adult roosters. The excretion of endogenous origin minerals viz., copper, magnesium, manganese and calcium was significantly (p<0.001) higher in caecectomized than in normal roosters. The difference in the endogenous excretion was 50; 60.45; 40.35 and 29.63 per cent for copper, magnesium, manganese and calcium, respectively, in caecectomized roosters. The caeca played a pivotal role in the reabsorption of endogenous origin calcium, magnesium, manganese and copper. The mechanism of phosphorus absorption by the caecal epithelium was negligible. The caecectomized roosters underestimated the bioavailability of copper in sunflower meal and manganese in almost all the test feedstuffs. The present investigation revealed that the caeca played a critical role in the absorption of minerals from vegetable protein feedstuffs which escape digestion and absorption in the small and large intestinal segments.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.287-292
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• 2018

Objective: The aim of present study was to assess the anti-oxidant potential of water-soluble peptides (WSPs) extract derived from buffalo and cow milk Cheddar cheeses at different stages of ripening. Methods: The antioxidant potential of WSPs extract was assessed through 2,2'-azinobis-3-ethylbenzothiazoline-6sulfonic acid (ABTS)-radical scavenging activity. In addition, impact of WSPs extract on cell viability and production of reactive oxygen species (ROS) in human colon adenocarcinoma Caco-2 (tert-butylhydroperoxide-induced) cell lines was also evaluated. Results: The ABTS-radical scavenging activity increased progressively with ripening period and dose-dependently in both cheeses. However, peptide extract from buffalo milk Cheddar cheese demonstrated relatively higher activity due to higher contents of water-soluble nitrogen. Intracellular ROS production in Caco-2 cells decreased significantly (p<0.05) till 150th day of cheese ripening and remained constant thereafter. Additionally, dose-dependent response of WSPs extract on antioxidant activity was noticed in the Caco-2 cell line. Conclusion: On the basis of current in vitro study, the Cheddar cheese WSPs extract can protect intestinal epithelium against oxidative stress due to their antioxidant activity.

• KSBB Journal
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• v.26 no.1
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• pp.62-68
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• 2011

To isolate Lactic acid bacteria for animals, we have screened from Kim-chi, swine intestine, swine feces, and dairy products by random selection and anti-viral, antipathogenic bacteria test. Among them, CLP-1 shown that inhibitory effect against rotavirus, porcine epidemic diarrhea (PED) virus, Salmonella sp, and E.coli. By examining biological property, API-ZYM and identified Lactobacillus plantarum by 16S rDNAgene sequence. CLP-1 determined resistance to low pH and bile salt. Futhermore, the cell body of CLP-1 adhered to the intestinal epithelium tissue of swine and Caco-2 cell. CLP-1 was examined on cell immune system modulating activity in vitro. The whole cell and cell culture supernatant was increasing of interferon-${\beta}$ activity. And then, CLP-1 increased prevention effect by Salmonella enteritidis infection in SPF chickens. And we determined similar result in pigs.