• Title/Summary/Keyword: mucosal protein

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Construction of a live attenuated Salmonella strain expressing FanC protein to prevent bovine enterotoxigenic Escherichia coli and evaluation of its immunogenicity in mice

  • Won, Gayeon;Kim, Hee Jung;Lee, John Hwa
    • Korean Journal of Veterinary Research
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    • v.57 no.1
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    • pp.9-15
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    • 2017
  • To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

Effect of Dietary Protein on the Serum Xanthine Oxidase Activity in Methanethiol-treated Rats (식이성 단백질 함량이 Methanethiol 투여한 흰쥐의 혈청 Xanthine Oxidase 활성에 미치는 영향)

  • 윤종국;전태원;임영숙
    • Journal of Environmental Health Sciences
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    • v.19 no.1
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    • pp.66-70
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    • 1993
  • Introduction : Methanethiol is a toxicant that is a byproduct in the industrial process (oil refinery), and it is produced in vivo from methionine via transamination in case of its overintake. And it also can be generated by the action of mucosal thiol Smethyltransferase on hydrogen sulfite which is formed by anaerobic bacteria in the intestinal tract. The toxicity of methanethiol has often been suggested as one of endogenous factors involved in the pathogenesis of hepatic encephalopathy. Furthermore, methanethiol could cause the membrane damage and inhibition of some membrane protective enzymes.

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Inhibitory Effects of ECQ on Indomethacin-Induced Gastric Damage in Rats

  • Jung, Juho;Nam, Yoonjin;Sohn, Uy Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.6
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    • pp.399-404
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    • 2012
  • We investigated inhibitory effects of extract containing quercetin-3-O-${\beta}$-D-glucuronopyranoside (ECQ) extracted from Rumex Aquaticus Herba on indomethacin-induced gastric damage in Rats. Gastritis was induced in male Sprague-Dawley rats (200~220 g) by oral administration of indomethacin at a dose of 40 mg/kg. One hour before administration of indomethacin, animals were orally pretreated with ECQ at doses of 0.3, 1, 3 or 10 mg/kg. Six hours after indomethacin administration, the rats were sacrificed and the stomach was excised and opened along the greater curvature, and the surface area of gastric lesion was measured using optical microscope. Superoxide dismutase (SOD), catalase (CAT), myeloperoxidase (MPO) activities and malondialdehyde (MDA) levels were measured by ELISA. Western blot analysis was performed to detect protein expression of SOD-2. Linear hemorrhagic mucosal lesions were observed in the stomach 6 hours after oral administration of indomethacin. Pretreatment with ECQ significantly reduced the severity of the lesions in a dose-dependent manner. It also inhibited the reductions in SOD and CAT activities and SOD expression by the indomethacin-induced gastric damage. In addition, the pretreatment with ECQ significantly suppressed the elevation of the MPO activity and the MDA levels induced by indomethacin. These results suggest that ECQ has the inhibitory effects via antioxidative action against indomethacin-induced gastritis in rats.

Vaccines for Prevention of Otitis Media and Pneumonia in Children (소아의 중이염 및 폐렴 예방을 위한 백신)

  • Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.16 no.1
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    • pp.13-23
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    • 2009
  • Acute otitis media (AOM) and pneumonia are among the most common infectious diseases of children. Both are mucosal infections and share many common features such as etiological agents, pathogenesis and immunity. Influenza plays an important role in the pathogenesis of AOM and pneumonia. A vaccine against influenza may have substantial impact on these diseases during the influenza season. In clinical trials, influenza vaccine has reduced the incidence of AOM and pneumonia complicating influenza in children. However, the efficacy of vaccines has been controversial in children less than 2 years of age. Similarly, vaccines against Streptococcus pneumoniae and Haemophilus influenzae type b (Hib), both common causes of AOM and pneumonia, have the potential to reduce the impact of disease. Clinical trials showed that the currently licensed 7-valent pneumococcal conjugate vaccine (PCV), administered during infancy, had an efficacy of 6-7% for the prevention of AOM, however, visits to the clinic for AOM were reduced by up to 20-30% after routine use in the U.S. Both Hib and PCVs have a proven effectiveness of >20% for prevention of radiologically confirmed pneumonia in children. The recently introduced pnuemococcal vaccine conjugated with protein D is expected to reduce AOM and pneumonia caused by non-typable H. influenzae, in addition to its effects on pneumococcal diseases. Considering their high incidence in children, recent achievements in the prevention of AOM and pneumonia with vaccines may have a significant economic and social impact.

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Effects of Korean Radish on DSS-Induced Ulcerative Colitis in Mice

  • Kim, Hyun-Kyoung
    • International Journal of Advanced Culture Technology
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    • v.6 no.4
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    • pp.97-108
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    • 2018
  • The present study aimed to investigate the comparative evaluation of pharmacological efficacy between sulfasalazine alone and combination with herbal medicine on dextran sodium sulfate (DSS)-induced UC in mice. Balb/c mice received 5% DSS in drinking water for 7 days to induce colitis. Animals were divided into five groups (n = 9): group I-normal group, group II-DSS control group, group III-DSS + sulfasalazine (30 mg/kg), group IV-DSS + sulfasalazine (60 mg/kg), group V-DSS + sulfasalazine (30 mg/kg) + Radish Extract mixture (30 mg /kg) (SRE). DSS-treated mice developed symptoms similar to those of human UC, such as severe bloody diarrhea and weight loss. SRE supplementation, as well as sulfasalazine, suppressed colonic length and mucosal inflammatory infiltration. In addition, SRE treatment significantly reduced the expression of pro-inflammatory signaling moleculesthrough suppression both mitogen-activated protein kinases(MAPK) and nuclear factor-kappa B ($NF-{\kappa}B$) signaling pathways, and prevented the apoptosis of colon. Moreover, SRE administration significantly led to the up-regulation of anti-oxidant enzyme including SOD and Catalase. This is the first report that Radish extract mixture combined with sulfasalazine protects against experimental UC via the inhibition of both inflammation and apoptosis, very similar to the standard-of-care sulfasalazine.

Monocytes Contribute to IFN-β Production via the MyD88-Dependent Pathway and Cytotoxic T-Cell Responses against Mucosal Respiratory Syncytial Virus Infection

  • Tae Hoon Kim;Chae Won Kim;Dong Sun Oh;Hi Eun Jung;Heung Kyu Lee
    • IMMUNE NETWORK
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    • v.21 no.4
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    • pp.27.1-27.12
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    • 2021
  • Respiratory syncytial virus (RSV) is the leading cause of respiratory viral infection in infants and children. However, little is known about the contribution of monocytes to antiviral responses against RSV infection. We identified the IFN-β production of monocytes using IFN-β/YFP reporter mice. The kinetic analysis of IFN-β-producing cells in in vivo RSV-infected lung cells indicated that monocytes are recruited to the inflamed lung during the early phase of infection. These cells produced IFN-β via the myeloid differentiation factor 88-mediated pathway, rather than the TLR7- or mitochondrial antiviral signaling protein-mediated pathway. In addition, monocyte-ablated mice exhibited decreased numbers of IFN-γ-producing and RSV Ag-specific CD8+ T cells. Collectively, these data indicate that monocytes play pivotal roles in cytotoxic T-cell responses and act as type I IFN producers during RSV infection.

Role of PKR and EGR-1 in Induction of Interleukin-S by Type B Trichothecene Mycotoxin Deoxynivalenol in the Human Intestinal Epithelial Cells (B형 트리코테센 곰팡이 독소 데옥시니발레놀에 의한 인체 장관 상피세포 염증성 인터루킨 8유도에서의 PKR과 EGR-1의 상호 역할 규명)

  • Park, Seong-Hwan;Yang, Hyun;Choi, Hye-Jin;Park, Yeong-Min;Ahn, Soon-Cheol;Kim, Kwan-Hoi;Lee, Soo-Hyung;Ahn, Jung-Hoon;Chung, Duk-Hwa;Moon, Yu-Seok
    • Journal of Life Science
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    • v.19 no.7
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    • pp.949-955
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    • 2009
  • Mucosal epithelia sense external stress signals and transmit them to the intracellular cascade responses. Ribotoxic stress-producing chemicals such as deoxynivalenol (DON) or other trichothecene mycotoxins have been linked with gastrointestinal inflammatory diseases by Fusarium-contamination. The purpose of this study was to test the hypothesis that DON evokes the epithelial sentinel signals of RNA-dependent protein kinase (PKR) and early growth response gene 1 (EGR-1), which together contribute to the pro-inflammatory cytokine interleukin 8 (IL-8) in human intestinal epithelial cells. PKR suppression by the dominant negative PKR expression attenuated DON-stimulated interleukin-8 production. Moreover, 1L-8 transcriptional activation by DON was also reduced by PKR inhibition in the human intestinal epithelial cells. Treatment with the PKR inhibitor also suppressed EGR-1 promoter activity, mRNA and protein induction, although mitogen-activated protein (MAP) kinases such as extracellular signal-regulated protein kinases (ERK) 1/2, p38, c-Jun N-terminal Kinase (INK) were little affected or even enhanced in presence of a PKR inhibitor. These patterns were also compared in the EGR-1-suppressed cells, which showed much more suppressed production of 1L-8. All things taken into consideration, DON-activated sentinel signals of EGR-1 via PKR mediated interleukin-8 production in human intestinal epithelial cells, which provide insight into the possible general mechanism associated with mucosal inflammation as an intestinal toxic insult by ribotoxic trichothecene mycotoxins.

LETM1 Promotes Gastric Cancer Cell Proliferation, Migration, and Invasion via the PI3K/Akt Signaling Pathway

  • Zhang, Yunfeng;Chen, Lele;Cao, Yifan;Chen, Si;Xu, Chao;Xing, Jun;Zhang, Kaiguang
    • Journal of Gastric Cancer
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    • v.20 no.2
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    • pp.139-151
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    • 2020
  • Purpose: Globally, there is a high incidence of gastric cancer (GC). Leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) is reported to play a vital role in several human malignancies. However, there is limited understanding of the role of LETM1 in GC. This study aims to investigate the effects of LETM1 on proliferation, migration, and invasion of GC cells. Materials and Methods: The expression levels of LETM1 in the normal gastric mucosal epithelial cells (GES-1) and GC cells were analyzed by quantitative real-time polymerase chain reaction and western blotting. CCK-8, wound healing, and Transwell invasion assays were performed to evaluate the effect of LETM1 knockdown or overexpression on the proliferation, migration, and invasion of the GC cells, respectively. Additionally, the effect of LETM1 knockdown or overexpression on GC cell apoptosis was determined by flow cytometry. Furthermore, the effect of LETM1 knockdown or overexpression on the expression levels of PI3K/Akt signaling pathway-related proteins was evaluated by western blotting. Results: The GC cells exhibited markedly higher mRNA and protein expression levels of LETM1 than the GES-1 cells. Additionally, the knockdown of LETM1 remarkably suppressed the GC cell proliferation, migration, and invasion, and promoted the apoptosis of GC cells, which were reversed upon LETM1 overexpression. Furthermore, the western blotting analysis indicated that LETM1 facilitates GC progression via the PI3K/Akt signaling pathway. Conclusions: LETM1 acts as an oncogenic gene to promote GC cell proliferation, migration, and invasion via the PI3K/Akt signaling pathway. Therefore, LETM1 may be a potential target for GC diagnosis and treatment.

A IMMUNOHISTOCHEMICAL STUDY ON HEAT SHOCK PROTEIN IN ORAL CARCINOGENESIS IN HAMSTER (햄스터 구강암 발생 과정에서 Heat Shock Protein에 관한 면역조직화학적 연구)

  • Choi, Kyu-Hwan;Lee, Dong-Keun;Kim, Eun-Chul;Jeong, Chang-Joo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.2
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    • pp.124-136
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    • 2001
  • Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor Immune response. Accordingly, the distribution and intensity of HSP70 and HSP47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and $90{\sim}120g$ were collected. 9,10-dimethyl -1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were race or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.

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A IMMUNOHISTOCHEMICAL STUDY ON HEAT SHOCK PROTEIN IN ORAL CARCINOGENESIS IN HAMSTER (햄스터 구강암 발생 과정에서 Heat Shock Protein에 관한 면역조직화학적 연구)

  • Choi, Kyu-Hwan;Lee, Dong-Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.20 no.4
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    • pp.362-372
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    • 1998
  • Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor immune response. Accordingly, the distribution and intensity of HSP 70 and HSP 47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and 90-120g were collected. 9,10-dimethyl-1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were rare or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP 47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.

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