• Title/Summary/Keyword: host cells

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Case study of HBV-related Disasters in a High-risk Family

  • Lee Gi Jun;Cho Jung Hyo;Cho Chong Kwan;Son Chang Gue
    • The Journal of Korean Medicine
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    • v.26 no.4
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    • pp.168-173
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    • 2005
  • Hepatitis B virus (HBV) is one of the most common intracellular parasites, of which 350 million people worldwide are chronic carriers. It also related to a high incidence of hepatocellular carcinoma. In general, it has been well known that HBV is a noncytolytic virus, so not the virus itself but any unfavorable response by host immune cells and inflammatory cytokines mainly result in chronic liver injury. From this viewpoint, we hopefully assume that Oriental therapies based on immunologic strategies may be able to provide a therapeutic alternative for caring for these illnesses. We also need to be thoroughly familiar with information about HBV epidemiology and the pathologic process of chronic HBV carriers. In this study, to clarify the important considerations of HBV infection and the high risk of HBV induced life-threatening diseases, we introduced our pilot practices given to the patients and the possibility of Oriental therapies as a novel strategy for chronic HBV carriers.

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Effect of Epigallocatechin-3-gallate on Expression of Chemokines in Human Nasal Mucosal Fibroblasts (Epigallocatechin-3-gallate의 사람 비점막 섬유아세포 케모카인발현에 대한 효과)

  • Cho, Jeong-Je;Leem, Kang-Hyun
    • Korean Journal of Pharmacognosy
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    • v.32 no.4 s.127
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    • pp.280-286
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    • 2001
  • Epigallocathechin-3-gallate (EGCG), the main polyphenol component in green tea, inhibits angiogenesis, urokinase, and matalloproteinases, and EGCG also has the antioxidative property. Recent reports proposed that EGCG may modulate the immune response on allergy or asthma. Human nasal mucosal fibroblasts are a rich source of cytokines, inflammatory mediators, and chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. The objective of this study was to investigate the effect of EGCG on the expression of the chemokines such as RANTES (regulated upon activation, normal T cell expressed and presumably secreted), eotaxin, and interleukin-8 (IL-8) in human nasal mucosal fibroblasts after stimulation with cytokines like IL-4, tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$, and $interferon-{\gamma}\;(IFN-{\gamma})$. To detect the expression of chemokine genes, RT-PCR was performed. Expressions of RANTES, eotaxin, and IL-8 mRNA stimulated with IL-4 and $TNF-{\alpha}$ were increased, respectively, while the expression of those genes incubated with $IFN-{\gamma}$ was similar pattern compared to control group. Analyses of chemokine genes of cells pretreated with EGCG showed that the expressions of eotaxin, and IL-8 genes stimulated $IFN-{\gamma}$ were higher compared with those not pretreated with EGCG.

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Anti-inflammatory and Antioxidant Effects of Clam Worm Extract Treated with Peptidoglycan (펩티도글리칸 처리된 갯지렁이 추출물의 항염증 및 항산화 효과)

  • Kim, Se-woong;Sapkota, Mahesh;Yang, Ming;Li, Liang;Soh, Yunjo
    • Korean Journal of Pharmacognosy
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    • v.48 no.3
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    • pp.187-194
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    • 2017
  • Peptidoglycan in inserts and mammals is well known to improve biological functions in the host's immune system. However, it is unclear how Peptidoglycan exerted its anti-inflammatory capacity especially in clam worm (Marphysa sanguinea). In this experiment, the anti-inflammatory and antioxidant effects of clam worm extract treated with (PCWE) peptidoglycan (Micrococcus luteus) in RAW264.7 cells were examined by measuring MDA, catalase, SOD, GSH-Px and inflammatory cytokines (nitric oxide, iNOS, interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$). PCWE significantly increased the activities of catalase, SOD and GSH-Px and decreased the level of MDA. Interestingly, PCWE induced activities of SOD and GSH-Px more than clam worm extract without peptidoglycan (CWE). In addition, PCWE decreased NO production, iNOS, COX-2, TNF-${\alpha}$ and IL-$1{\beta}$ better than CWE. Taken together, these results indicate that PCWE has the potential as a natural antioxidant and a therapeutic for inflammation-related diseases.

Structural Characteristics of Immunostimulating Polysaccharides from Lentinus edodes

  • Lee, Hee-Hwan;Lee, Jong-Seok;Cho, Jae-Yeol;Kim, Young-Eon;Hong, Eock-Kee
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.455-461
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    • 2009
  • There is a significant amount of experimental evidence suggesting that polysaccharides from mushrooms enhance the host immune system by activating various mechanisms in immune cells, including macrophages. In this study, polysaccharides from Lentinus edodes were found to stimulate the functional activation of macrophages to secrete inflammatory mediators and cytokines and increase the phagocytotic uptake. The chemical properties of the stimulatory polysaccharides, CPFN-G-I, CPBN-G, and CPBA-G, were determined based on their monosaccharide composition, which mainly consisted of glucose and mannose. According to FT-IR and GC/MS, the structure of CPFN-G-I, purified from the fruiting body of L. edodes, was found to consist of a $\beta$-1,6-branched-$\beta$-1,4-glucan, whereas CPBN-G and CPBA-G, purified from the liquid culture broth, were found to be composed of a heteromannan. The configuration of the p-linkage and triple helical conformation of each polysaccharide were confirmed using a Fungi-Fluor kit and Congo red, respectively.

Electron Microscopy Studies on the Formation of Polyhedra Occlusion Bodies of Autographa californica Nuclear Polyhedrosis Virus (미생물 살충제인 Autographa californica Nuclear Polyhydrosis Virus의 Polyhydra 형성 과정의 전자현미경적 연구)

  • Lee Hyung-Hoan
    • Applied Microscopy
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    • v.11 no.1
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    • pp.51-57
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    • 1981
  • The process of the formation of polyhedra occlusion bodies and occlusion of viral nucleocapsids of Autographa californica Nuclear Polyhedrosis Virus in Spodoptera frugiperda cell were photomicrographed and described. Progeny viral nucleocapsids were observed in the nuclei of the host cells, bundled and then enveloped. The nucleoapsids were mainly accumulated near the membrane-like profiles. The nuclear membrane were hypertophied up to the cytoplasmic membrane. Prepolyhedral bodies were observed and they were growing with the accumulations of thread-like materials(polypeptides) produced by viral genes. The bundled and enveloped nucleocapsids were occluded into the growing polyhedra.

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Herpes Zoster Ophthalmicus in a Patient with Malignant Lymphoma (악성림프종 환자에서 발생한 안부 대상포진)

  • Lee, Jun-Hak;Kim, Hyung-Tae;Park, Jun-Beom;Park, Sang-Cheol;Kwon, Young-Eun
    • Journal of Hospice and Palliative Care
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    • v.8 no.1
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    • pp.52-56
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    • 2005
  • Herpes zoster (HZ) is an acute infection of the unilateral sensory dermatome caused by varicella-zoster virus (VZV) and is characterized by vesicular eruption and unilateral pain along the involved dermatome. Although the pathogenesis of HZ is incompletely understood, it is thought that when cell-mediated immunity falls below a critical level, dormant VZV within cells of the sensory ganglia are allowed to replicate and infect the host with the resultant clinical presentation of HZ. It has been associated with immunosuppressed states, such as advanced age, leukemia, lymphoma, chemotherapy and/or radiation treatment. We present a case of a 62-year-old female patient with malignant lymphoma suffering herpes zoster ophthalmicus who did not respond to conventional treatment, and in whom the application of various nerve blocks and patient-controlled analgesia produced moderate pain relief. The patient died twenty days later due to cardiopulmonary failure.

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A Microbial D-Hydantoinase is Stabilized and Overexpressed as a Catalytically Active Dimer by Truncation and Insertion of the C-Terminal Region

  • KIM, GEUN-JOONG;HAK-SUNG KIM
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.242-248
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    • 2002
  • Previously, it was reported that the nonhomologous C-terminal regions of the D-hydantoinases are nonessential for catalysis, but affect the oligomeric structure of the enzyme [3]. In an effort to further confirm the above observation, the C-terminal region-inserted enzyme was constructed by attaching a peptide (22 residues) at the C-terminal of the D-hydantoinase from Bacillus thermocatenulatus GH2, and its structural and biochemical properties were compared with both the wild-type and C-terminal region-truncated enzymes. As a result, native tetrameric D-hydantoinase was dimerized as the truncated enzyme, and the inserted mutant with a new sequence was expressed as a catalytically active form in E. coli. Expression level of the inserted and truncated enzymes were found to be significantly increased compared to the level of the wild-type enzyme, and this appears to be due to the reduced toxic effect of the mutant enzymes on host cells. Dimerized enzymes exhibited increased thermo- and pH stabilities considerably when compared with the corresponding wild-type enzyme. Comparison of the substrate specificity between the mutant and wild-type enzymes suggests that the substrate specificity of the D-hydantoinase is closely linked with the oligomeric structure.

Cloning and Expression of Serratia marcescens Protease Gene in Escherichia coli

  • KIM, MYUNG-HEE;SOO-KEUN CHOI;BON-TAG KOO;BYUNG-SIK SHIN;CHEON-BAE SOHN;JEONG-IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.2 no.4
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    • pp.231-236
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    • 1992
  • A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and $45^\circ{C}$, respectively.

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Biochemical Characterization of an ABC Transporter Gene Involved in Cephabacin Biosynthesis in Lysobacter lactamgenus

  • Park, Myoung-Jin;Yon, Jei-Oh;Lim, Si-Kyu;Ryu, Dewey D.-Y.;Nam, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.635-638
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    • 2004
  • An ATP-binding-cassette (ABC) transporter gene in the cephabacin biosynthetic gene cluster of Lysobacter lactamgenus was characterized. The amplified orf10 (cpbJ) gene was subcloned into pET-28a(+) vector and expressed in E. coli BL21(DE3) strain by 0.5 mM IPTG at $30^{\circ}C$. The membrane fraction of recombinant E. coli cells was separated by ultracentrifugation, and solubilized using 2.5% octyl-$\beta$-D-glucoside. Using the solubilized membrane fraction, the artificial proteoliposomes were reconstituted and analyzed for the biological activity of CpbJ protein. Upon measuring ATPase activity, the proteoliposome made from recombinant E. coli membrane proteins showed slightly higher activity than that from host E. coli membrane proteins. In the measurement of membrane transport activity, the reconstituted proteoliposome of recombinant E. coli membrane proteins exhibited higher activity when both substrates of cephalosporin C and L-Ala-L-Ser were applied, compared to the case of cephalosporin C or L-Ala-L-Ser only. It implies that the CpbJ protein is an ABC transporter secreting cephabacin antibiotics synthesized in cytoplasm.

Immunostimulatory Effects of ${\beta}$-glucan Purified from Paenibacillus polymyxa JB115 on Mouse Splenocytes

  • Kim, Ji-Mi;Joo, Hong-Gu
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.4
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    • pp.225-230
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    • 2012
  • We investigated the effects of ${\beta}$-glucan purified from Paenibacillus polymyxa JB115 on the viability and proliferation of splenocytes. Splenocytes play a critical role in host immunity. MTT assays and trypan blue exclusion tests revealed that ${\beta}$-glucan significantly promoted the viability and proliferation of splenocytes over a range of concentrations. However, there was no specific subset change. ${\beta}$-glucan protected splenocytes from cytokine withdrawal-induced spontaneous cell death. For further mechanistic studies, ELISA assay revealed that ${\beta}$-glucan enhanced the expression of anti-apoptotic molecules and interleukin 7 (IL-7), a cytokine critical for lymphocyte survival. We also investigated the IL-2 dependency of ${\beta}$-glucan-treated splenocytes to determine if treated cells could still undergo clonal expansion. In flow cytometric analysis, ${\beta}$-glucan induced increased levels of the activation marker CD25 on the surface of splenocytes and ${\beta}$-glucan-treated splenocytes showed higher proliferation rates in response to IL-2 treatment. This study demonstrates that ${\beta}$-glucan can enhance the survival of splenocytes and provides valuable information to broaden the use of ${\beta}$-glucan in research fields.