• Title/Summary/Keyword: host cells

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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • 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,

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The Establishment of Nitrogen Fixation by Cultured Cell-Rhizobium Association Through Tissue Culture Technique in Soybean (대두조직배양세포(大豆組織培養細胞) - Rhizobium에 의(依)한 질소고정력(窒素固定力))

  • Kang, Sang Jae;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.27-35
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    • 1986
  • This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

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Definition of the peptide mimotope of cellular receptor for hepatitis C virus E2 protein using random peptide library (Random peptide library를 이용한 C형 간염바이러스 E2 단백질 세포막 수용체의 peptide mimotope 규명)

  • Lee, In-Hee;Paik, Jae-Eun;Seol, Sang-Yong;Seog, Dae-Hyun;Park, Sae-Gwang;Choi, In-Hak
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.77-86
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    • 2001
  • Background: Hepatitis C virus(HCV), a family of Flaviviridae, has a host cell-derived envelope containing a positive-stranded RNA genome, and has been known as the maj or etiological agent for chronic hepatitis, hepatic cirrhosis, and hepatocellular carcinoma. There remains a need to dissect a molecular mechanism of pathogenesis for the development of therapeutic and effective preventive measure for HCV. Identification of cellular receptor is of central importance not only to understand the viral pathogenesis, but also to exploit strategies for prevention of HCV. This study was aimed at identifying peptide mimotopes inhibiting the binding of E2 protein of HCV to MOLT-4 cell. Methods: In this study, phage peptide library displaying a random peptides consisting of 7 or 12 random peptides was employed in order to pan against E2 protein. Free HCV particles were separated from the immune complex forms by immunoprecipitation using anti-human IgG antibody, and used for HCV-capture ELISA. To identify the peptides inhibiting E2-binding to MOLT-4 cells, E2 protein was subj ect to bind to MOLT-4 cells under the competition with phage peptides. Results: Several phage peptides were selected for their specific binding to E2 protein, which showed the conserved sequence of SHFWRAP from 3 different peptide sequences. They were also able to recognize the HCV particles in the sera of HCV patients captured by monoclonal antibody against E2 protein. Two of them, showing peptide sequence of HLGPWMSHWFQR and WAPPLERSSLFY respectively, were revealed to inhibit the binding of E2 protein to MOLT-4 cell efficiently in dose dependent mode. However, few membrane-associated receptor candidates were seen using Fasta3 programe for homology search with these peptides. Conclusion: Phage peptides containing HLGPWMSHWFQR and WAPPLERSSLFY respectively, showed the inhibition of E2-binding to MOLT-4 cells. However, they did not reveal any homologues to cellular receptors from GenBank database. In further study, cellular receptor could be identified through the screening of cDNA library from MOLT-4 or hepatocytes using antibodies against these peptide mimotopes.

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EXPRESSION OF THE GENES OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND VON WILLEBRAND FACTOR IN SALIVARY GLAND TUMORS (타액선 종양에서 혈관내피성장인자와 von Willebrand 인자 유전자 발현에 관한 연구)

  • Chung, Ji-Hun;Kim, Ji-Hyuk;Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.41-51
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    • 2008
  • Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumor which compromises about 6$\sim$8% of all tumors followed by the adenoid cystic carcinoma (ACC) and adenocarcinoma. Most deaths from salivary carcinomas are caused by recurrent or metastatic lesions that are resistant to conventional therapy. Therefore, knowledge of cellular properties and tumor-host interactions that influence the vascular metastasis is important for the design of more effective therapy of salivary carcinomas. Neoangiogenesis is essential for tumor growth, which is postulated to be fundamentally dependent on the induction of stromal neovascularization. However, how neovascularization takes place in live tissue has not been fully established, especially in recruitment and differentiation of endothelial cells in the salivary gland tumors. Vascular endothelial growth factor (VEGF) is a heparin-binding, dimeric polypeptide growth factor known to exert its mitogenic activity specifically on endothelial cells. VEGF has been shown th be directly involved in angiogenesis, which in essential for the pathogenesis of many solid tumors. von Willebrand factor (vWF) is a large multimeric protein synthesized by megakaryocytes and endothelial cells that enable platelets to adhere to exposed subendothelium and, as well, to respond to changes in the blood flow. Recent studies suggest that increased levels of vWF correlate with progression of disease, metastasis, or survival time and thus may have a prognostic significance. vWF is explained as an acute phase proteins which is increased in cancer or as a result of increased endothelial cell synthesis associated with tumor-induced angiogenesis. Due to adhesive properties of vWF, its increased concentrations may also contribute metastasis of tumor. In this study, we determined the mRNA expression of VEGF and vWF in salivary ACC, MEC and pleomorphic adenoma by in situ hybridization. As a result, stronger expression of VEGF and vWF was seen in salivary ACC and MEC which has more invasive nature than the salivary benign tumor.

Development of Probiotic Products and Challenges (프로바이오틱 제품 개발 동향과 과제)

  • Seo, Jae-Gu;Lee, Gwa-Soo;Kim, Jin-Eung;Chung, Myung-Jun
    • KSBB Journal
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    • v.25 no.4
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    • pp.303-310
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    • 2010
  • Probiotics beneficially affect the health of the host via various mechanisms in the intestine. Recent developments in probiotic products have mainly been made to maximize probiotic effects in human. In this regard, probiotic products containing doubly coated or encapsulated cells, multi-species probiotics, or high viable cell number (1010 viable cells/gram or more) have been developed and are already available in the market. Until now, the majority of probiotics contain live cells but little attention has been paid to other alternative products such as heat-killed cell or bacteriocin-containing ones, which could have broad applications due to advantages over live cell-based probiotics, such as safety and stability. In addition, genetically engineered lactic acid bacteria could be of great importance in the field of alimentary health if they are carefully designed for biological safety. Although a number of probiotics are marketed by claiming health benefits, regulations for health claims will be more stringent. Therefore sufficient scientific and clinical evidences supporting the safety and efficacy of the potential probiotic strain will be required by the regulatory authority for a health claim, which thus may have a huge impact on the future probiotic market.

Immune Function-enhancing Effects of Bojungikkitanggami-bang (보중익기탕가미방의 면역기능 증진 효과)

  • Lee Sang Hun;Lee Seung Eon;Lee Si Hyeong;Shin Jo Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.2
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    • pp.528-533
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    • 2004
  • The immune system acts to protect the host from infectious agents that exist in the environment and from other noxious insults. The immune system has two functional divisions: the innate and the acquired. Both components involve various factors such as cytokines. A number of methodologies exist to assess aspects of immune function. There are large inter-individual variations in many immune functions even among the healthy. Genetics, age, gender, smoking habits, habitual levels of exercise, alcohol consumption, diet, stage in the female menstrual cycle, stress, history of infections and vaccinations, and early life experiences are likely to be important contributors to the observed variation. While it is clear that individuals with immune responses significantly below 'normal' are more susceptible to infectious agents and exhibit increased infectious morbidity and mortality, it is not clear how the variation in immune function among healthy individuals relates to variation in susceptibility to infection. Oriental medicine is an important factor contributing to immune competence. The author investigated the immune enhancement effects of Bojungikkitanggami-bang (BITB). The forced swimming test (FST) has been used as a screening model for new immune enhancement agents. In the present study, the author investigated the effects of BITB on FST and blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactate dehydrogenase (LDH); creatinine; and total protein (TP). The author found that BITB (1 g/kg) significantly reduced the immobility time in the FST compared to the control. In addition, the contents of Glc, LDH, BUN, TP in the blood serum were increased in BITB (1g/kg)-fed group. Also, the author investigated the effects of BITB on the production of cytokines in human T-cell line, MOLT-4 cells. BITB (1 mg/ml) significantly increased the interferon (IFN)-vproduction compared with media control (about 2.2-fold for IFN-γ) at 24 h. However, BITB has not affect the production of IL-2 and IL-4. In addition, BITB increased the protein expression level of IFN-γ in MOLT-4 cells. Thus, BITB may have therapeutic value in generating or enhancing immune function in a clinical setting.

Strength of the Mutant Promoters for the \beta-xylosidase gene of Bacillus stearothermophilus No. 236 (Bacillus stearothermophilus No. 236 \beta-xylosidase 유전자 변이 Promoter의 Strength분석)

  • 최용진;김미동
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.111-116
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    • 2003
  • The xylA gene of Bacillus stearothermophilus No. 236 encoding $\beta$-xylosidase was cloned and sequenced previously. The transcriptional start site of the xylA gene cloned in E. coli was identified to be the guanine (G) by primer extension analysis. This supports that the expression of xylA gene is also directed in the E. coli cells by the previously determined transcription initiation signals, -10 sequence (CATAAT) and -35 sequence (TTGTTA) separated by 12 bp. To increase the expression of $\beta$-xylosidase, firstly the spacer region of xylA promoter was extended from 12 to 17 bp, and then the -10 and -35 elements were converted into their respective consensus sequences. The mutant promoters thus obtained were tested for their activities in both the E. coli and B. subtilis host cells. The change of the length of the spacer region from 12 to 17 bp resulted in a 1.6- and 2.5-fold increase in promoter strength in comparison with the wild type promoter in E. coli and B. subtilis cells, respectively. Also, strength of the promoter with the fourth T to A transversion on its -35 element increased in the transcription level by about 35 times compared with that of wild-type promoter. However, surprisingly the 5' end C-to-T transition of the -10 hexamer showed a 5- to 15-fold reduction in $\beta$-xylosidase activity in both E. coli and B. subtilis. Together, the present data demonstrated that the 5' end nucleotide C of the -10 sequence CATAAT and the fourth nucleotide A of the -35 hexamer are two most critical nucleotides for the promoter activity in the context of the xylA promoter.

Hematopoietic stem cell transplantation in children with acute leukemia: similar outcomes in recipients of umbilical cord blood versus marrow or peripheral blood stem cells from related or unrelated donors

  • Yi, Eun-Sang;Lee, Soo-Hyun;Son, Meong-Hi;Kim, Ju-Youn;Cho, Eun-Joo;Lim, Su-Jin;Cheuh, Hee-Won;Yoo, Keon-Hee;Sung, Ki-Woong;Koo, Hong-Hoe
    • Clinical and Experimental Pediatrics
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    • v.55 no.3
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    • pp.93-99
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    • 2012
  • Purpose: This study compared outcomes in children with acute leukemia who underwent transplantations with umbilical cord blood (UCB), bone marrow, or peripheral blood stem cells from a human leukocyte antigen (HLA)-matched related donor (MRD) or an unrelated donor (URD). Methods: This retrospective study included consecutive acute leukemia patients who underwent their first allogeneic hematopoietic stem cell transplantation (HSCT) at Samsung Medical Center between 2005 and 2010. Patients received stem cells from MRD (n=33), URD (n=46), or UCB (n=41). Results: Neutrophil and platelet recovery were significantly longer after HSCT with UCB than with MRD or URD ($p$ <0.01 for both). In multivariate analysis using the MRD group as a reference, the URD group had a significantly higher risk of grade III to IV acute graft-versus-host disease (GVHD; relative risk [RR], 15.2; 95% confidence interval [CI], 1.2 to 186.2; $p$=0.03) and extensive chronic GVHD (RR, 6.9; 95% CI, 1.9 to 25.2; $p$ <0.01). For all 3 donor types, 5-year event-free survival (EFS) and overall survival were similar. Extensive chronic GVHD was associated with fewer relapses (RR, 0.1; 95% CI, 0.04 to 0.6; $p$ <0.01). Multivariate analysis showed that lower EFS was associated with advanced disease at transplantation (RR, 3.2; 95% CI, 1.3 to 7.8; $p$ <0.01) and total body irradiation (RR, 2.1; 95% CI, 1.0 to 4.3; $p$=0.04). Conclusion: Survival after UCB transplantation was similar to survival after MRD and URD transplantation. For patients lacking an HLA matched donor, the use of UCB is a suitable alternative.

Adhesion Ability and Inhibition of Enterohemorrhagic E. coli O157:H7 Adhesion to Intestinal Epithelial Cells in Lactobacillus acidophilus (Lactobacillus acidophilus의 장 상피세포에 대한 부착능력 및 장 출혈성 대장균의 부착 억제 능력)

  • 김영훈;박순옥;한경식;오세종;유승권;김세헌
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.86-91
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    • 2004
  • The ability of probiotics containing Lactobacillus acidophilus to adhere to the intestinal epithelium may play an important role in colonization of the gastrointestinal tract and preventing enteric pathogen such as enterohemorrhagic E. coli(EHEC O157:H7. In the study, we investigated the adhesion to human intestinal epithelial cells(HT-29) of strains of L. acidophilus(3 from human, 2 from pig, and 1 from calf). All of the tested strains of L. acidophilus were highly observed adhesion ability(from 10$\^$6/ to 10$\^$7/ cfu/mL), compared to L. rhamnosus GG as control. Also, adhered strains of L. acidophilus were significantly preserved in serial wash-out steps. However, no correlation could be observed between cell surface hydrophobicity and adhesion abilities of the tested strains of L. acidophilus. Inhibition of adhesion of EHEC O157:H7 was also examined, a 2 log cycle reduction was observed by all of the tested strains of L. acidophilus. These results suggest that the strains of L. acidophilus with high adhesion ability are resistant to wash-out and adhesion ability inhibition by selected strains of L. acidophilus helps to prevent adhesion of EHEC O157:H7 to intestinal epithelial cells.

Studies on Viruses Isolated from Lilium spp. In Korea I. Broad Bean Wilt Virus and Cucumber Mosaic Virus (한국산 나리류에서 분리한 바이러스에 관한 연구 I. Broad Bean Wilt Virus 및 Cucumber Mosaic Virus)

  • Chang Moo Ung;Chung Jae Dong
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.223-235
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    • 1987
  • Samples showing yellow mosaic symptom of Lilium spp. with necrotic fleck, stunting, malformation, and colour breaking were collected from lily-growing areas in the southern part of Korea. Two viruses were distinguished under a electron microscope and their host range, serological reaction, stability in sap, type of aphid transmission, and relations with cells and tissues were examined. Broad bean wilt virus (BBWV) was transmitted by sap-inoculation to 23 plant species in 8 families and by the aphid, Myzus persicae. This virus was inactivated after 10 min at 70C, at dilution of $10^{-3}$, and after 6 days at about 20C. Electron microscopic examination of purified preparation showed that the virus is spherical particle of 28nm in diameter. The virus reacted positively with BBWV-antiserum in agar gel diffusion test. In ultrathin sections of BBWV infected tissues, large aggregates or crystalline array of virus particles and vesicular body were found in the cytoplasm, vacuole, and nucleus of mesophyll cells. Cucumber mosaic virus (CMV) was transmitted by sap-inoculation. Electron microscopic examination of its purified preparation showed spherical particles of 30nm in diameter. The virus reacted positively with CMV-Y strain-antiserum in agar gel diffusion test. In ultrathin sections of CMV infected tissues, crystalline array of virus particles were found in the vacuole and a large number 0f virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells. When each of these viruses was retransmitted to Lilium tigrinum. L. concolor, and L. auratum, BBWV induced slight symtoms and colour breaking, but CMV induced yellowing mosaic or necrotic fleck.

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