• Title/Summary/Keyword: Plaque assay

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A Study on Serologic Diagnosis for Dengue Virus Infection

  • Sang-Wook Park;Je-Hoon Yang;Hyung-Joon Bae;Hi-Joo Moon;Young-Dae Woo
    • Biomedical Science Letters
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    • v.8 no.4
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    • pp.269-273
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    • 2002
  • Dengue fever (DF) is an acute febrile illness caused by dengue viruses in the family Flaviviridae, genus Flavivirus. DF has so far posed any problem in Korea, however it has been recently believed to be associated with oversea's traveler infected with dengue virus. Antibody titers of sera from DF patients against dengue virus were measured by indirect immunofluorescence assay (IFA) and plaque reduction neutralization test (PRNT), including the haematologic test. Three of patients with DF showed highly fluorescent and neutralizing antibody titers by IFA and PRNT assay. Two of them showed higher, remarkably. Meanwhile, one of them was tested and resulted in severe tirombocytopenia, elevated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) activities as well as mild leucopenia, increased monocytes and basophils and depressed lymphocytes in haematological differential count.

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Effect of Fumonisin B1 on the Bacterial Virus Multiplication (세균 바이러스 증식에 대한 Fumonisin B1의 영향)

  • 이길수
    • Toxicological Research
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    • v.12 no.1
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    • pp.17-20
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    • 1996
  • The effect of Fumonisin B1, a mycotoxin produced by Fusarium moniliforme on bacterial viruses P1 and Lambda, was investigated by the virus plaque assay. Fumonisin B1 inhibited the P1 viral multiplication in the concentration range from $100{\mu}g$/ml to $400{\mu}g$/ml. The inhibition was Fumonisin B1 concentration-dependent. Another bacterial virus Lambda multiplication was also inhibited by lower concentration of Fumonisin B1 ($10{\mu}g$/ml~$50{\mu}g$/ml). This inhibition was dependent on Fumonisin B1 and on virus-Fumonisin B1 reaction time. Sensitivity of bacteriophage Lambda to Fumonisin B1 was higher than that of P1 virus. Lambda vital DNA was treated in vitro with Fumonisin B1 at various concentration. Significant DNA fragmentation by Fumonisin 191 was observed in the agarose gel electrophoresis. Lambda viral DNA was partially digested even in the Fumonisin B1 $10{\mu}g$ and the level of its fragmentation was dependent on Fumonisin B1 amount up to $30{\mu}g$ per assay.

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Effect of Glycyrrhizae Radix on the Immune Responses(I) - Immuno-regulatory Action of 50% Methanol Extract - (감초가 면역반응에 미치는 영향 (I) - 50% 메탄올 엑스의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.154-164
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    • 1991
  • These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

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Enhancement of Type A Macrophage Scavenger Receptor Expression by Ginsenoside Rg3 in Rat Microglia (흰쥐 뇌 소교세포에서 진세노사이드 Rg3의 Type A Macrophage Scavenger Receptor 발현 증진효과)

  • Joo, Seong-Soo;Hwang, Kwang-Woo;Lee, Do-Ik
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.147-150
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    • 2005
  • Macrophage scavenger receptors (MSRs) induce microglial interaction with ${\beta}$-amyloid fibrils (fA${\beta}$) that are associated with Alzheimer's disease (AD). Although microglia are know n to have a dual effect on formation of plaque and clearance of fA${\beta}$ in the AD brain, receptor-mediated phagocytosis is a very important tool for preventing amyloid plaque via activated microglia in the early stage of AD. In the study, we examined whether ginsonoside Rg3 enhances the microglial Phagocytosis of A${\beta}$1-42 through Phagocytosis assay, gene expression (RT-PCR) and protein assay (western blots) for the cell responsiveness presented between Rg3-treated and non-treated groups. Fluro-labeled Ac-LDL and E.coli particles were used as control proteins for phagocytosis. In previous studies, this was a particularly interesting property of Rg3 in the stimulation and phagocytosis of macrophages in the periphery. We report here that ginsenoside Rg3 increased the expression of type-A MSR (MSR-A) in microglia and thus accelerated the phagocytosis with an effective degradation of engulfed fA${\beta}$. This result suggests that Rg3 may play an important role in removing fA${\beta}$ by enhancing the receptor-mediated phagocytosis. In addition, Rg3 could be a potential candidate for balancing the rate of production of fA${\beta}$ in AD brain.

LIGHT is Expressed in Foam Cells and Involved in Destabilization of Atherosclerotic Plaques through Induction of Matrix Metalloproteinase-9 and IL-8

  • Kim, Won-Jung;Lee, Won-Ha
    • IMMUNE NETWORK
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    • v.4 no.2
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    • pp.116-122
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    • 2004
  • Background: LIGHT (TNFSF14) is a member of tumor necrosis factor superfamily and is the ligand for TR2 (TNFRSF14/HVEM). LIGHT is known to have proinflammatory roles in atherosclerosis. Methods: To find out the expression pattern of LIGHT in atherosclerotic plaques, immunohistochemical analysis was performed on human carotid atherosclerotic plaque specimens. LIGHT induced atherogenic events using human monocytic cell line THP-1 were also investigated. Results: Immunohistochemical analysis revealed expression of LIGHT and TR2 in foam cell rich regions in the atherosclerotic plaques. Double immunohistochemical analysis further confirmed the expression of LIGHT in foam cells. Stimulation of THP-1 cells, which express TR2, with either recombinant LIGHT or immobilized anti-TR2 monoclonal antibody induced interleukin-8 and matrix metalloproteinase(MMP)-9. Electrophoretic mobility shift assay demonstrated that LIGHT induces nuclear localization of transcription factor, nuclear factor $(NF)-{\kappa}B$. LIGHT induced activation of MMP-9 is mediated by $NF-{\kappa}B$, since treatment of THP-1 cells with the $NF-{\kappa}B$ inhibitor PDTC (pyrrolidine dithiocarbamate) completely blocked the activation of MMP-9. Conclusion: These data indicate that LIGHT is expressed in foam cells in atherosclerotic plaques and is involved in atherogenesis through activation of pro-atherogenic cytokine IL-8 and destabilization of plaque by inducing matrix degrading enzyme.

Effects of Phellinus linteus Extracts on the Humoral Immune Respons in Normal and Cyclophosphamide-treated Mice (상황버섯 추출물이 정상 마우스와 cyclophosphamide로 처리된 마우스의 체액성 면역기능에 미치는 영향)

  • 표명윤;현수미;양기숙
    • Biomolecules & Therapeutics
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    • v.9 no.3
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    • pp.194-200
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    • 2001
  • Phellinus linteus (PL)-hot water extract (PL-W) or- methanol extract (PL-M) was orally administered alone (single dose of 400, 800, 1600 mg/kg; 800 mg/kg/day for 5 days) or with cyclophosphamide (CY, 20 mg/kg, i.p.) to female ICR mice. Within PL alone-treated group, WBC and plaque forming cells (PFC) to SRBC were slightly and significantly enhanced when compared with control group. The relative thymus and spleen weights, WBC and PFC numbers were significantly decreased by the treatment of CY, whereas those values were markedly increased by the concomitant treatment of CY and PL when compared with CY administration alone. To assess the effects of PL and/or CY on the mitogen response of splenocytes io LPS, mouse splenocytes were stimulated with or without LPS in the presence of various concentration of PL and/or CY in vitro and splenocytes proliferation (SP) was measured by MTT assay. PL alone increased both SP and LPS- stimulated SP. Moreover, SP and LPS-induced SP suppressed by the treatment of CY alone were significantly restored by PL-treatment. These activities were higher by PL-M than by PL-W, These results indicated that PL was able to increase humoral immunity and to inhibit immunotoxicity induced by CY.

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Antiviral Effect of Amphotericin B on Japanese Encephalitis Virus Replication

  • Kim, Hun;Kim, Seong-Jun;Park, Sue-Nie;Oh, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.121-127
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    • 2004
  • Amphotericin B (AmB), an amphipathic polyene macrolide, is an antifungal drug produced by Streptomyces nodosus. Recently, AmB has been shown to exert antiviral activity against rubella virus and human immunodeficiency virus by different mechanisms. In this study, we evaluated the antiviral effect of AmB against Japanese encephalitis virus (JEV) and investigated which step of the viral life cycle was inhibited by AmB to understand the mechanism of antiviral action of AmB. AmB reduced both plaque size and number in the infected cells in a dose-dependent manner. In addition, a 200-fold reduction of infectious virus titer was observed by treatment of infected cells with $5\mug/ml$ of AmB. AmB acted at the post virus-infection step, but not during adsorption of virus to host cells. Western blot analysis revealed that the accumulated level of JEV envelope protein dramatically decreased in the infected cells by treatment with $5-10\mug/ml$ of AmB. Our results indicate that AmB inhibits the replication of JEV at the postinfection step by interfering with viral replication and/or by inhibiting the synthesis of viral proteins.

Immune Suppression and Stimulation of Antioxidants -Effect of Propyl gallate on Murine Humoral Immune Functions and Methemoglobin Content- (한산화제의 면역억제 및 항진 연구 - 항산화제인 Propyl gallate가 체액성 면역기능과 Aniline 유도 Methemoglobin 함량에 미치는 영향 -)

  • 유충규;황미경
    • Journal of Food Hygiene and Safety
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    • v.3 no.2
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    • pp.83-88
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    • 1988
  • Propyl gallate used as an antioxidant was examined for its effects on murine Immune system and metbemoglobin content treated with anillne. As immunotoxicology assay parameters, we adopted circulating leukocytes and immunoorgan weights for pathtoxicology, IgM plaque forming cells and Artbus reaction for humoral immunity. Propyl gallate's effects were observed as follows; 1. Propyl gallate decreased circulating leukocyte counts, dose dependently. 2. Relative immunoorgan weigbts were not affected. 3. Propyl gallate diminisbed IgM PFCs/spleen cell and IgM PFCs/spleen. 4. Propyl gallate decreased Arthus reaction. 5. Propyl gallate did not affect metbemogiobin content treated wltb aniIIne.

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PCR based Detection of Helicobacter spp. in Saliva, Dental plaque, Vomitus and Feces of Dogs

  • Ha, Sung-Woo;Yoo, Jong-Hyun;Chung, Tae-Ho;Jung, Woo-Sung;Youn, Hwa-Young;Chae, Joon-Seok;Hwang, Cheol-Yong
    • Journal of Veterinary Clinics
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    • v.25 no.6
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    • pp.447-451
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    • 2008
  • This study was aimed to determine the prevalence of Helicobacter spp. in privately owned pet dog's oral cavity samples (saliva, dental plaque, vomitus) and fecal samples in Korea and to evaluate the potential route for transmission. Total 100 patients dogs attending one Veterinary Medical Teaching Hospital were examined by Helicobacter genus-specific PCR assay and these dogs were divided into two groups whether they had gastrointestinal signs (vomiting, nausea and diarrhea) or not. The total detection rate of Helicobacter spp. by PCR in saliva, dental plaque and fecal samples was 23%, 1% and 68% respectively. The difference of prevalence with regarding the gastrointestinal sings was not significant. In vomitus, two of seven samples had positive results. These results suggested that Helicobacter spp. are present in the oral cavity although they were present in very low number and are not like to be normal oral flora of the oral cavity and Helicobacter spp. in dogs could be transmitted through oral-oral, gastrooral and fecal-oral route.

A Collaborative Study on Korean Standard JE Vaccine for Potency Assay

  • Kim, Jae-Ok;Shin, Jin-Ho;Baek, Sun-Young;Min, Kyung-Il;Min, Bok-Soon;Ryu, Seung-Rel;Kim, Byoung-Guk;Kim, Do-Keun;Ahn, Mi-Jin;Park, Mi-Kyung;Song, Hye-Won;Lee, Chung-Keel;Lee, Seok-Ho;Park, Sue-Nie
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.745-750
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    • 2004
  • The objective of this collaborative study was to establish a Korean standard of Japanese encephalitis (JE) vaccine (mouse brain-derived, formalin-inactivated) for potency assay. A candidate preparation proposed as a Korean standard was provided by GreenCross Vaccine, and six laboratories, including one national control laboratory and five manufacturers of JE vaccine, participated in the study. The potency of the candidate preparation and a reference standard obtained from Japan was estimated by mouse immunogenicity assay using the in vitro plaque reduction neutralization test (PRNT). The results of 72 assays conducted by the 6 laboratories showed that the overall mean potency estimate of the candidate preparation was 2.455 log median plaque reduction neutralization antibody titer per 0.5-ml dosage administered twice in mice at 7-day intervals, and that the mean potency ratio of the candidate preparation relative to the reference standard was 1.074. The potency estimates were quite variable among laboratories irrespective of the preparation. The variability of assays assessed by Z scores and coefficient of variation (CV) were in general within the level of acceptance (Z scores within $\pm3$ and $CV\;\leq\;15%$). Therefore, we concluded that the candidate preparation would be suitable as a national standard for testing the potency of JE vaccine (inactivated).