• Title/Summary/Keyword: Hepatitis E

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Development of Human Antibody Inhibiting RNase H Activity of Polymerase of Hepatitis B Virus Using Phage Display Technique (Phage Display 기법을 이용한 B형 간염 바이러스 Polymerase의 RNase H 활성을 억제하는 인간 단세포군 항체의 개발)

  • Lee, Seong-Rak;Song, Eun-Kyoung;Jeong, Young-Joo;Lee Young-Yi;Kim, Ik-Jung;Choi, In-Hak;Park, Sae-Gwang
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.16-22
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    • 2004
  • Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.

Infection Status of Hospitalized Diarrheal Patients with Gastrointestinal Protozoa, Bacteria, and Viruses in the Republic of Korea

  • Cheun, Hyeng-Il;Cho, Shin-Hyeong;Lee, Jin-Hee;Lim, Yi-Young;Jeon, Ji-Hye;Yu, Jae-Ran;Kim, Tong-Soo;Lee, Won-Ja;Cho, Seung-Hak;Lee, Deog-Yong;Park, Mi-Seon;Jeong, Hye-Sook;Chen, Doo-Sung;Ji, Yeong-Mi;Kwon, Mi-Hwa
    • Parasites, Hosts and Diseases
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    • v.48 no.2
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    • pp.113-120
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    • 2010
  • To understand protozoan, viral, and bacterial infections in diarrheal patients, we analyzed positivity and mixedinfection status with 3 protozoans, 4 viruses, and 10 bacteria in hospitalized diarrheal patients during 2004-2006 in the Republic of Korea. A total of 76,652 stool samples were collected from 96 hospitals across the nation. The positivity for protozoa, viruses, and bacteria was 129, 1,759, and 1,797 per 10,000 persons, respectively. Especially, Cryptosporidium parvum was highly mixed-infected with rotavirus among pediatric diarrheal patients (29.5 per 100 C. parvum positive cases), and Entamoeba histolytica was mixed-infected with Clostridium perfringens (10.3 per 100 E. histolytica positive cases) in protozoan-diarrheal patients. Those infected with rotavirus and C. perfringens constituted relatively high proportions among mixed infection cases from January to April. The positivity for rotavirus among viral infection for those aged $\leq$ 5 years was significantly higher, while C. perfringens among bacterial infection was higher for $\geq$ 50 years. The information for association of viral and bacterial infections with enteropathogenic protozoa in diarrheal patients may contribute to improvement of care for diarrhea as well as development of control strategies for diarrheal diseases in Korea.

Apoptotic effect of formaldehyde in cultured human hepatocyte cell lines (인간 간세포주 에서 포름알데히드에 의한 세포 사멸 효과)

  • Park, Soo-Hyun
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.462-467
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    • 2009
  • Exposure of formaldehyde (FA), one of the major compounds in pesticides and in the onset of sick house syndrome, has been implicated in the development of diverse diseases. Liver is a very important organ to body metabolism and drug detoxification. Apotosis of hepatocytes is associated with the onset of liver diseases such as hepatitis. However, the apoptotic effect of FA in hepatocytes is not clear. Therefore, this study was conducted to investigate the effect of FA on the apoptosis in HepG2 cells, a human hepatocyte cell line. As a result, FA (> $500\;{\mu}M$) decreased cell viability and increased lactate dehydrogenase activity in HepG2 cells, which was blocked by the treatment of vitamin E and N-acetylcysteine (NAC). In addition, FA decreased glutathione (GSH) contents and Bcl-2 levels, while increasing lipid peroxide formation and Bax levels. It also cleaved caspase-3 form, which was blocked by the treatment of vitamin E and NAC. It is insisted that FA induced apoptosis via oxidative stress in human hepatocytes.

Activation of Toll-like receptor 9 and production of epitope specific antibody by liposome-encapsulated CpG-DNA

  • Kim, Dong-Bum;Kwon, Hyung-Joo;Lee, Young-Hee
    • BMB Reports
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    • v.44 no.9
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    • pp.607-612
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    • 2011
  • Several investigators have shown that CpG-DNA has outstanding effects as a Th1-responsive adjuvant and that its potent adjuvant effects are enhanced by encapsulation with a liposome of proper composition. In this study, we showed that encapsulation with phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE): cholesterol hemisuccinate (CHEMS) complex enhances the immunostimulatory activity of CpG DNA and the binding of CpG-DNA to TLR9. We also examined involvement of myeloid differentiation protein (MyD88) and NF-${\kappa}B$ activation in liposome-encapsulated CpG-DNA-induced IL-8 promoter activation. In this manuscript, the natural phosphodiester bond CpG-DNA encapsulated by DOPE : CHEMS complex is designated as Lipoplex(O). Importantly, we successfully screened B cell epitopes of envelope protein (E protein) of hepatitis C virus (HCV-E) and attachment glycoprotein G of human respiratory syncytial virus (HRSV-G) by immunization with complexes of several peptides and Lipoplex(O) without carriers. Therefore, Lipoplex(O) is potentially applicable as a universal adjuvant for peptide-based epitope screening and antibody production.

Increased DNA Polymerase Fidelity of the Lamivudine Resistant Variants of Human Hepatitis B Virus DNA Polymerase

  • Hong, Young-Bin;Choi, Yong-Wook;Jung, Gu-Hung
    • BMB Reports
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    • v.37 no.2
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    • pp.167-176
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    • 2004
  • Although efficient antiviral lamivudine is used for HBV-infected patients, a prolonged treatment with nucleoside analogs often results in lamivudine-resistant variants. In this study, we evaluated the fidelity of the lamivudine-resistant variants. The FLAG-tagged wild-type (FPolE) and Met550 variants (FPolE/M550A, M550V, and M550I) of HBV DNA polymerases were expressed in insect cells then purified. Like many other reverse transcriptases, no $3'{\rightarrow}5'$ exonuclease activity was detected in the HBV DNA polymerase. Since there is no proofreading activity, then the use of the site-specific nucleotide misincorporation method is beneficial. From the $f_{ins}$ value analysis, it is evident that M550I and M550V exhibit higher fidelity values than the wild-type HBV DNA polymerase, while M550A exhibits similar fidelity values. It is therefore suggested that lamivudine resistance comes from the stringency to dNTP binding and the discrimination of dCTP and lamivudine in M550V and M550I.

Acute Organophosphorus Pesticide Poisoning (급성 유기인계 농약 중독)

  • Lee, Mi-Jin;Park, Joon-Seok;Hong, Tai-Yong;Park, Sung-Soo;You, Yeon-Ho
    • Journal of The Korean Society of Clinical Toxicology
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    • v.6 no.2
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    • pp.83-90
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    • 2008
  • Organophosphate (OP) pesticides are the most common source of human toxicity globally, causing high mortality and morbidity despite the availability of atropine as a specific antidote and oximes to reactivate acetylcholinesterase. The primary toxicity mechanism is inhibition of acetylcholinesterase (AchE), resulting in accumulation of the neurotransmitter, acetylcholine, and abnormal stimulation of acetylcholine receptors. Thus, the symptoms (muscarinic, nicotinic, and central nervous system) result from cholinergic overactivity because of AchE inhibition. OP can also cause rhabdomyolysis, pancreatitis, parotitis, and hepatitis. OP therapy includes decontamination, supportive therapy, and the use of specific antidotes such as atropine and oximes. However, there has been a paucity of controlled trials in humans. Here we evaluated the literature for advances in therapeutic strategies for acute OP poisoning over the last 10 years.

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Value Addition Span of Silkworm Cocoon - Time for Utility Optimization

  • Reddy, R. Manohar
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.1
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    • pp.109-113
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    • 2008
  • Realizing the scope of utilizing by-products of silk cocoons by applying appropriate methods is the immediate crave to optimize returns. The nutritive value of pupae suits for human diet, feed for poultry, carps, fish, rabbits, piggery and dogs. The pupal skin, fat, oil, cocoon palade have applications in oleo chemical, soap, glycerin, cosmetic, artificial fibres, membranes and n-triacontanol isolation. The pupal proteins Chitin, Shinki fibroin, Serrapeptidase, glucosamine are latent precursors of post surgical, anti-carcinogenic, anti-inflammative, anti-bacterial, anti-histaminic, gastric, hepatitis, pancreatitis, leukocytopenia, neurological, ophthalmic, blood pressure, cardiac and diabetic medicines and for preparation of vitamins A, E and K. The silk and its proteins sericin and fibroin are potentially used for wound healing, diabetes, impotence, sinusitis, arthritis, edema, cystitis, epididymitis, tissue regeneration, cancer, post-surgical trauma and used as anti-oxidatives, bio-adhesives, ultra violet screens and bio-active textiles. The waste cocoons can be used in making art crafts like garlands, carpets, overcoats, decoratives and greeting cards. The in-depth research towards utility optimization and make aware this reality to sericulturists, reelers, weavers, traders, entrepreneurs, policy makers etc., is the upright want of the today's Sericulture industry.

A New Coumestan Glucoside from Eclipta prostrata

  • Seo, Young Ju;Kil, Hyun Woo;Rho, Taewoong;Yoon, Kee Dong
    • Natural Product Sciences
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    • v.26 no.4
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    • pp.289-294
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    • 2020
  • Eclipta prostrata is an annual herb, belonging to Asteraceae family, and has been traditionally used to improve immunity and treat hepatitis and bacterial disease in Korea. In this study, a new coumestan glucoside (1) along with ten known compounds (2 - 11) was isolated from E. prostrata. The chemical structures of isolates were elucidated to be wedelolactone-9-O-β-D-glucopyranoside (1), wedelolactone (2), demethylwedelolactone (3), apigenin (4), apigenin-7-sulfate (5), luteolin (6), luteolin-7-sulfate (7), luteolin-7-O-β-D-glucopyranoside (8), pratensein-7-O-β-D-glucopyranoside (9), 3,4-di-O-caffeoylquinic acid (10) and 3,5-di-O-caffeoylquinic acid (11) based on the spectroscopic evidence.

Effects of anti-inflammation and cell protection through biphenyl dimethyl dicarboxylate on Rat Microglia

  • Joo, Seong-Soo;Kang, Hee-Chul;Lee, Do-Ik
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.132.1-132.1
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    • 2003
  • Biphenyl dimethyl dicarboxylate (DDB) is a by-product produced in process of synthesizing Schizandrin-C. Generally, DDB has known to protect hepatocytes and to decrease the index of liver enzyme (e.g. GOT and GPT) in chronic hepatitis. The present study was aimed to demonstrate whether DDB can protect the brain cell, especially the Alzheimer brain in vitro. As Alzheimers disease can be induced by activated microglia, a macrophage in the brain, through Abeta peptide (A$\beta$) produced from amyloid precursor protein (APP). (omitted)

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The Levels of IgG Subclasses and Vaccine-induced Viral Antibodies in Childhood Minimal Change Nephrotic Syndrome (소아 신증후군에서 IgG 아군 및 바이러스 항체 값)

  • Song Yong-Hoon;Lee Kyung-Yil;Kim Dong-Un;Koh Dae-Kyun;Lee Byung-Churl
    • Childhood Kidney Diseases
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    • v.9 no.2
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    • pp.143-148
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    • 2005
  • Purpose : Hypogammaglobulinemia has been observed in nephrotic syndrome, but its pathophysiology remains unknown. We evaluated serum immunoglobulins, IgG subclasses, and vaccine-induced viral antibodies(anti-hepatitis B surface IgG and anti-measles IgG) in children with minimal change nephrotic syndrome(MCNS). Methods : Using the stored sera, the levels of immunoglobulin(IgC, IgM, IgA, and IgC) and IgG subclasses(IgG 1, 2, ,3, and 4), anti-HBs Ab and anti-measles IgG of 21 children with MCNS were analyzed and compared to those of 25 age-matched healthy children. Results : The mean values of IgG and IgG1 were $390{\pm}187\;mg/dL$ and $287{\pm}120\;mg/dL$ in nephrotic children, and $1,025{\pm}284\;mg/dL$ and $785{\pm}19\;mg/dL$ in control children, respectively. The values of the total IgG and the 4 IgG subclasses in nephrotic children were all significantly depressed(P<0.001), but the IgM($251{\pm}183\;mg/dL\;vs. 153{\pm}55\;mg/dL$, P=0.02) and IgE values(P=0.01) were elevated, and the IgA values were not changed. The seropositivity of anti-HBs IgG was 42.9$\%$(9 of 21 cases) in the MCNS group and 52$\%$(13/25) in the control group, and that of anti-measles IgG was 75$\%$(16/21) and 92$\%$(23/25), respectively, but there was no statistical difference between the two groups. Conclusion : IgG and IgG subclass levels in MCNS children are all depressed without significant seronegativity of the vaccine-induced viral antibodies. Further studies are needed to resolve the cause of hypogammaglobulinemia in MCNS. (J Korean Soc Pediatr Nephrol 2005;9:143-148)

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