• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1401-1407
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• 2009

Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.

• Tuberculosis and Respiratory Diseases
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• v.53 no.2
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• pp.196-201
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• 2002

Malignant melanoma is a highly malignant form of cutaneous cancer derived from melanocytes. The lesion frequently metastasizes to the lymph nodes, lung, liver and bone. However, an endobronchial metastasis and a primary malignant. melanoma of the lung are quite rare. We report a case of an unknown primary malignant melanoma with a pulmonary and endobronchial metastasis in a 34 years old male. He complained of coughing and black-colored sputum. Abnormal skin and mucosal lesions were not found during a physical examination. A chest X-ray revealed multiple nodular masses in both lung fields. A flexible bronchoscopy showed two yellowish small nodules at the entry of left lower bronchus. Vimentin, the S-100 protein, and HMB-45 stain positive melanoma cells were detected at the bronchoscopic biopsy specimen.

• Korean Journal of Poultry Science
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• v.50 no.2
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• pp.109-118
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• 2023

Four-d-old broiler chicks were randomly assigned to 3 groups with 9 replicates (8 birds/cage) under high ambient temperature; birds fed a basal diet (CON), a basal diet supplemented with 0.25% of probiotic complex (LPB, 1 × 10⁶ Lactobacillus plantarum, 1 × 10⁶ Bacillus subtilis, and 1 × 10⁶ Saccharomyces cerevisiae) and 0.5% probiotic complex (HPB). Immediately after 28-d feeding trial, 6 birds having average body weight per group were sacrificed for evaluating the effects of probiotics on antioxidant parameters in the serum, intestine, and liver of birds. As results, serum biochemical parameters of nitrogen components including total protein, albumin, urea nitrogen, and glutathione were unaffected by dietary probiotics. In addition, serum superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione S-transferase (GST) activities, and lipid peroxidation (MDA) were not changed by dietary probiotic supplement in birds. In the intestinal mucosa, SOD activity in the HPB group significantly (P<0.05) increased compared with that in the CON and the LPB groups. Lipid peroxidation in the HPB group significantly (P<0.05) decreased compared with that in the CON group. However, there was no statistical difference in GPX, and GST activities in the intestinal mucosa among treatment groups. In the liver, the activities of SOD, GPX, and GST, and the level of MDA were unaffected by probiotic supplement. In conclusion, 0.5% of probiotics significantly increased SOD activity and decreased lipid peroxidation in the intestinal mucosa, suggesting that probiotic complex could be potential to improve the small intestinal antioxidant capacity of bird under high ambient temperature conditions.

• Animal Bioscience
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• v.35 no.10
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• pp.1461-1478
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• 2022

The maintenance of poultry gut health is complex depending on the intricate balance among diet, the commensal microbiota, and the mucosa, including the gut epithelium and the superimposing mucus layer. Changes in microflora composition and abundance can confer beneficial or detrimental effects on fowl. Antibiotics have devastating impacts on altering the landscape of gut microbiota, which further leads to antibiotic resistance or spread the pathogenic populations. By eliciting the landscape of gut microbiota, strategies should be made to break down the regulatory signals of pathogenic bacteria. The optional strategy of conferring dietary fibers (DFs) can be used to counterbalance the gut microbiota. DFs are the non-starch carbohydrates indigestible by host endogenous enzymes but can be fermented by symbiotic microbiota to produce short-chain fatty acids (SCFAs). This is one of the primary modes through which the gut microbiota interacts and communicate with the host. The majority of SCFAs are produced in the large intestine (particularly in the caecum), where they are taken up by the enterocytes or transported through portal vein circulation into the bloodstream. Recent shreds of evidence have elucidated that SCFAs affect the gut and modulate the tissues and organs either by activating G-protein-coupled receptors or affecting epigenetic modifications in the genome through inducing histone acetylase activities and inhibiting histone deacetylases. Thus, in this way, SCFAs vastly influence poultry health by promoting energy regulation, mucosal integrity, immune homeostasis, and immune maturation. In this review article, we will focus on DFs, which directly interact with gut microbes and lead to the production of SCFAs. Further, we will discuss the current molecular mechanisms of how SCFAs are generated, transported, and modulated the pro-and anti-inflammatory immune responses against pathogens and host physiology and gut health.

• The Korean Journal of Zoology
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• v.17 no.2
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• pp.57-68
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• 1974

These experiments were performed in order to study histologically and histochemically on the epithelial cells of gall bladder in Carassius carassius, Bufo bufo gargarizans, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus. The results of the observation were as follows: 1. There were different cell types in the epithelium of gall bladder in each animal and it could not be supported histochemically that the epithelia cells of gall bladder were divided into two cell types of the rod-shaped and barrel-shaped ones. 2. The epithelium of gall bladder in Carassius carassius, Bufo bufo gargarizans, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus was simple columnar epithelium. 3. The eosinophilities of cytoplasm in the epithelial cells of gall bladder were in uniform stronger in the upper portion of nucleus in Carassius carassius, Bufo bufo gargarizans and Natrix tigrina lateralis than its other portions, and in Urloncha striata var. domesticus and Lepus cuniculus var. domesticus existed uniformly in all portions, but there were many non-eosinophilic cells in Bufo bufo gargarizans and many cells that weakly eosinophilic around nucleus in Bos taurus var. domesticus. 4. The periodic acid Schiff's reactivities in the epithelial cells of gall bladder were different in each other and the epithelial cells in PAS reaction were divided into two cell types of the dark and light ones. There presented the light cells of 6.4%, 4.3% and 3.7% of epithelial cell of gall bladder in Carassius carassius, Bufo bufo gargarizans and Urloncha striata var. domesticus for each other, but were not presented in Natrix tigrina lateralis and Bos taurus var. domesticus. 5. The ninhydrin-Schiff-active proteins were much in the epithelial cells of gall bladder in Bos taurus var. domesticus, Carassius carassius, Urloncha striata var. domesticus and Natrix tigrina lateralis in order and were much in epithelial cells in the upper portion of mucosal folds in Carassius carassius, Urloncha striata var. domesticus and Natrix tigrina lateralis, and the ninhydrin-Schiff-active protein of the epithelium of gall bladder in Bufo bufo gargarizans was uniformly distributed. 6. The epithelial cells of gall bladder in Carassius carassius, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus had no stain reactivity or weak stain reactivity to neutral fat and all epithelial cells in Bufo bufo gargarizans had strong stain reactivity, though they were different in quantity of epithelial cell portion. 7. The stain reactivities to RNA and DNA were stronger in the epithelial cells of the upper portion of mucosal fold than in those of other portions.

• Applied Microscopy
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• v.39 no.2
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• pp.117-124
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• 2009

Clonorchis sinensis is the most important widely distributed parasite of the human bile duct in East Asia and the most prevalent parasitic helminth in Korea. The prevalence rate of human clonorchiasis has remained at about 2.9% in Korea. C. sinensis induces dilatation of the duct, hyperplasia of the mucosa, metaplasia or neoplasia of the mucosal epithelium, periductal inflammation and fibrosis, and thickening of the ductal wall. Fibroblast are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may also contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. In this study, ultrastructural changes, the distribution of lectin receptors and actin protein in cultured SD rat bile duct fibroblast after infection of C. sinensis were observed. Experimental group had been divided into four groups: normal bile duct fibroblast cultured in basal media (G1); C. sinensis infected bile duct fibroblast cultured in basal media (G2); normal bile duct fibroblast cultured in basal media containing excretory-secretory product (ESP) (G1-1); C. sinensis infected bile duct fibroblast cultured in basal media containing ESP (G2-1). Overall, once a host is infected by C. sinensis, it affects the host to the extent that sialic acid of ductal fibroblast is increased. Number of cytoplasmic process of SD rat bile duct fibroblast was increased. Actin protein and sialic acid were located in cell surface. Fibroblast induced by C. sinensis was not recovered to normal fibroblast. The cytoplasm bulk and cytoplasmic process were increased whereas the growth rate of the fibroblast of infected SD rat was reduced rather than that of normal fibroblast. In result, it inhibits fibroblast proliferation and increases actin protein on fibroblast cytoplasm, and so causes fibroblast metamorphosis and cellular mutation.

• 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.

• Korean Journal of Veterinary Research
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• v.48 no.4
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• pp.441-449
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• 2008

The genus Lactobacillus is the largest of the genera included in lactic acid bacteria and is associated with mucosal membranes of human and animal. Only a few Lactobacillus plasmid-encoded functions have been discovered and used. In this study, a novel plasmid (pILR091) was isolated from a wild L. reuteri isolated from pig and described the characteristics of its replicons, genetic organization, and relationship with other plasmids. After digestion of the plasmid, pILR091, with SalI, plasmid DNA was cloned into the pQE-30Xa vector and sequenced. The complete sequence was confirmed by the sequencing of PCR products and analyzed with the Genbank database. The isolate copy number and stability were determined by quantitative-PCR. The complete sequence of L. reuteri contained 7,185 nucleotides with 39% G-C content and one cut site by two enzymes, SalI and HindIII. The similar ori sequence of the pC194- rolling circle replication family (TTTATATTGAT) was located 63 bp upstream of the protein replication sequence, ORF 1. Total of five ORFs was identified and the coding sequence represented 4,966 nucleotides (70.4%). ORF1 of pILR091 had a low similarity with the sequence of pTE44. Other ORFs also showed low homology and E-values. The average G-C content of pILR091 was 39%, similar with that of genomic DNA. The copy number of pILR091 was determined at approximately 24 to 25 molecules per genomic DNA. These results suggested that pILR091 might be a good candidate to construct a new vector, which could be used for cloning and expression of foreign genes in lactobacilli.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.502-510
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• 2009

Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A protein from rice to direct the yeast-expressed LTB towards the endoplasmci reticulum to oligomerize with the expected pentameric size. The expression and assembly of the recombinant LTB were confirmed in both the cell-free extract and culture media of the recombinant strain using a Western blot analysis. The binding of the LTB pentamers to intestinal epithelial cell membrane glycolipid receptors was further verified using a GM1-ganglioside enzyme-linked inmmunosorbent assay (GM1-ELISA). On the basis of the GM1-ELISA results, pentameric LTB proteins comprised approximately 0.5-2.0% of the total soluble proteins, and the maximum quantity of secreted LTB was estimated to be 3 mg/l after a 3-day cultivation period. Consequently, the synthesis of LTB monomers and their assembly into biologically active aligomers in a recombinant S. cerevisiae strain demonstrated the feasibility of using a GRAS microorganism-based adjuvant, as well as the development of carriers against mucosal disease.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.37-44
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• 2015

Objectives : The present study was designed to evaluate the anti-inflammatory and anti-oxidative stress activities through regulation of Nrf2-mediated genes by Rhei rhizoma and Glycyrrhiza rhizoma combined extract (RGE) in reflux esophagitis.Methods : The antioxidant activity of RGE in vitro was measured in terms of radical scavenging capacity such as DPPH and ABTS. RGE was administered at 350 mg/kg body weight prior to induction of reflux esophagitis. Reflux esophagitis was induced that tied the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats.Results : RGE scavenged DPPH and ABTS effectively and IC₅₀of RGE each were 4.9 μg/ml and 45.6 μg/ml. Our results show that RGE administration markedly ameliorated mucosal damage upon histological evaluation. In serum and esophagus tissue, RGE significantly suppressed the oxidative stress biomarkers. Reflux esophagitis induced rats exhibited down-regulation of antioxidant-related proteins in the esophagus; however, the levels with treatment of RGE were significantly higher than those of vehicle reflux esophagitis rats. RGE treatment caused significant reductions in activation of NF-κB transcription factor. Thus, RGE significantly exhibited potent anti-inflammatory activities by suppressing the protein expression levels of pro-inflammatory proteins such as COX-2 and iNOS and inflammatory cytokines such as TNF-αin the esophagus tissue.Conclusions : Reflux esophagitis caused considerable levels of oxidative stress in the esophageal mucosa and the administration of RGE reduced the esophageal mucosa damage through the regulation of Nrf2 and NF-κB pathways. Our findings can considered as supplementary therapy in the prevention or treatment of reflux esophagitis.