• Title/Summary/Keyword: virus assay

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Difference in the Transcriptional Activity of the Interleukin-4 Promoter Haplotypes (Interleukin-4 유전자의 Promoter 일배체형에 따른 전사능의 차이)

  • Choi, Eun Hwa;Kim, Hee Sup;Chanock, Stephen J.;Lee, Hoan Jong
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.495-499
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    • 2005
  • Purpose : Interleukin-4(IL-4) is a critical component of the Th2 cytokine pathway and contributes to severity of respiratory syncytial virus(RSV) bronchiolitis. Previous studies observed an association between severe RSV bronchiolitis in Korean children with a common haplotype of the IL4 promoter. This study was performed to investigate functional differences of the variant IL4 promoter haplotypes. Methods : Genomic DNA was obtained from 20 children from 6 to 48 months of age in the Department of Pediatrics, Seoul National University Bundang Hospital. The IL4 promoter spanning an 1.2 kb region was amplified and haplotype was determined by cloning and the PHASE reconstruction. Transcriptional activity of Jurkat T cells which were transfected with each IL4 haplotype were analyzed by use of luciferase assay. Results : Three haplotypes of the IL4 promoter have been identified with the frequency of GCC(7 percent), TCC(17 percent), and TTT(76 percent). The TTT haplotype demonstrated the highest luciferase values in both unstimulated and PMA-stimulated Jurkat T cells. Increases in transcriptional activity compared to GCC have been shown in TTT(5.3 fold higher) followed by TCC(4.2 fold higher) in unstimulated Jurkat T cells. Conclusion : We provided evidence that increased transcriptional activity of the TTT haplotype of the IL4 promoter, which has previously been over-represented in Korean children with severe RSV bronchiolitis. Therefore, IL-4 could play a potential role in the pathogenesis of RSV infection, possibly via an altered transcriptional activity of the different IL4 haplotypes.

Environmental Control Accomodative to Ecosystem on the Prevention of Nuclear Polyhedrosis Virus Disease in the Silkworm. Bombyx mori (생태친화적(生態親和的) 사육환경(飼育環境) 제어방식(制御方式) 확립(確立)을 위한 누에 핵다각체병(核多角體病) 발생요인(發生要因) 분석(分析))

  • Han, Myung-Sae;Lim, Jong-Sung
    • Current Research on Agriculture and Life Sciences
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    • v.15
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    • pp.69-81
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    • 1997
  • For the prevention of worldwide prevalent disease of nuclear polyhedrosis virus (NPV), environmental conditions and their incidence of grasserie was investigated through 57 cases of silkworm rearing from the year of 1979 to 1993 in the countries of Korea, Japan, and Philippines. Relationship between the occurrence of NPV and environmental factors were also analysed from the aspect of causal pathogenesis. Unfavorable foactors related to the prevalence of NPV disease was reconfirmed by the assay of experimental rearing. Silkworms reared on mulberry leaves or artificial diet appeared similar result on the occurrence of grasserie. Disinfection by formalin and simple sweeping or washing was not significantly different on the occurrence of NPV disease. Following insufficient ventilation on the younger larvae. from the 1st to 3rd instar, the disease by NPV at the later stage was remarkably emphasized those insidence. An experimental rearing from 1993 to 1996 demonstrated the prevention of NPV disease by simple cleaning of sweeping under the condition of air forced ventilation, the customal practice of disinfection with formalin or any other chemical agents could be omissible.

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Detection of HHV6 and EBV in histiocytic necrotizing lymphadenitis (Histiocytic necrotizing lymphadenitis에서 HHV6과 EBV의 검출)

  • Park, Kyung Hee;Park, Sung Shik;Kim, Ji Yeon;Park, Su Eun
    • Clinical and Experimental Pediatrics
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    • v.51 no.9
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    • pp.987-991
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    • 2008
  • Purpose : Kikuchi-Fujimoto disease (KFD), also known as histiocytic necrotizing lymphadenitis (HNL), is a self-limited disease characterized by cervical lymphadenopathy and fever. The etiology of KFD remains unknown; however, the self-limiting nature of HNL suggests the cause of this disease could be viral infection. For this reason, several viruses have been evaluated as possible etiologies of HNL, including Epstein-Barr virus (EBV), human herpesvirus 6 (HHV6), human herpesvirus 8 (HHV8), and cytomegalovirus (CMV). The aim of this study was to examine the relationship of EBV and HHV6 to HNL. Methods : Data pertaining to 51 cases with biopsy-confirmed HNL were collected between January 1999 and December 2005, from the Department of Pathology, College of Medicine, Pusan National University, Busan, Korea. The clinical records-including data regarding age, gender, duration of fever, and lymph node involvementwere reviewed retrospectively. The in situ hybridization (ISH) assay was performed by EBER PNA probe (Dako, Capinteria, CA, USA), and immunohistochemistry testing was performed with anti-HHV type 6 monoclonal antibodies (Chemicon, Temecula, CA, USA). Results : The HNL patients in this study were 24 males and 27 females, ranging in age from seven to 61 years (median: 25.9). ISH for EBV was positive in 8/51 (15.7%) biopsies, and immunohistochemistry for HHV6 was positive in 15/51 (29.4%) biopsies. Serologic analysis of EBV IgM was performed in 23 cases; only one patient was positive for EBV IgM and EBV ISH. Conclusion : Our study could not provide supportive evidence of a viral pathogenesis for HNL; therefore, cases of HNL may not have a dominant viral cause. However, some rare exceptional cases may have been caused by viral infection.

Phospholipase D Activity is Elevated in Hepatitis C Virus Core Protein-Transformed NIH 3T3 Mouse Fibroblast Cells (C형 간염바이러스의 core 단백질에 의해 암화된 쥐의 섬유아세포에서 phospholipase D 효소활성의 증가)

  • Kim, Joonmo;Jung, Eun-Young;Jang, Kyung-Lib;Min, Do-Sik
    • Journal of Life Science
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    • v.13 no.5
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    • pp.551-558
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    • 2003
  • Hepatitis C Virus (HCV) is associated with a severe liver disease and increased frequency in the development of hepatocellular carcinoma. Overexpression of HCV core protein is known to transform fibroblast cells. Phospholipase D (PLD) activity is commonly elevated in response to mitogenic signals, and PLD has been also reported to be overexpressed and hyperactivated in some human cancer. The aim of this study was to understand how PLD can be regulated in HCV core protein-transformed NIH3T3 mouse fibroblast cells. We observed that in unstimulated state, basal PLD activity was higher in NIH3T3 cells overexpressing HCV core protein than in vector-transfected cells. Although expression of PLD and protein kinase C (PKC) in core protein-transformed cells was similar with that of control cells, phorbol 12-myristate 13-acetate (PMA), which is known to activate PKC, stimulated significantly PLD activity in core protein-transformed cells, compared with that of the control cells. PLD activity assay using PKC isozyme-specific inhibitor, and PKC translocation experiment showed that PKC-$\delta$ was mainly involved in the PMA-induced PLD activation in the core-transformed cells. Taken together, these results suggest that PLD might be implicated in core protein-induced transformation.

Disease monitoring of Alaska pollock (Gadus chalcogrammus) based on growth stages (명태 (Gadus chalcogrammus)의 성장 단계별 질병 모니터링)

  • Kim, Kwang Il;Byun, Soon-Gyu;Kang, Hee Woong;Nam, Myung-Mo;Choi, Jin;Yoo, Hae-Kyun;Lee, Chu
    • Korean Journal of Ichthyology
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    • v.29 no.1
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    • pp.62-68
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    • 2017
  • The Alaska pollock (Gadus chalcogrammus) belongs to the family Gadidae; it is a cold water fish, and has been developed as a novel aquaculture species in Korea. In this study, we describe ongoing surveillance for aquatic animal pathogens based on growth stages. We investigated bacterial flora in rearing water, and monitored pathogens; we also analyzed histopathological traits of abnormal fish. In rearing water, the total bacterial counts were $2.1{\times}10^3cfu/mL$ and Vibrio spp. (52%) were predominant in the larvae stage. In the juvenile and adult stages, the total bacterial counts were $3.4{\times}10^3$ and $3.2{\times}10^2cfu/mL$, respectively (with Pseudomonas sp. as the predominant species; 90% and 52%). This result revealed that the bacterial flora in rearing water changed depending on the feeding types. No virulent-bacteria or problematic viruses (VHSV, viral hemorrhagic septicemia virus; NNV, nervous necrosis virus; MBV, marine birnavirus) were detected from outwardly healthy fish using either culture or PCR assay. Some juveniles (less than 5%) had gas bubbles on the gill lamellae, degeneration of the corneal epithelium, and choroid gland degeneration, suggesting that these symptoms were caused by external injury and secondary infection by opportunistic bacteria. Disease management is important to cope with disease emergence in the novel aquaculture species Alaska pollock.

Comparison of Three Third-Generation Anti-HCV Enzyme Immunoassay Tests (세가지 효소면역측정 시약을 이용한 C형 간염 바이러스 항체 검사의 비교)

  • Cho, Hee-Soon;Moon, Jin-Young;Lee, Chae-Hoon;Kim, Kyung-Dong
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.143-150
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    • 1998
  • The aim of this study was to evaluate domestic enzyme immunoassay(EIA) kit 'LG RCD 3.0' (LG) for the detection of antibody to hepatitis C virus(anti-HCV) in comparision with Axsym HCV version 3.0(Axsym), Cobas Core anti-HCV EIA(Cobas). Cobas kit shows better clear distinction between positive and negative by signal/cutoff ratio(S/C), but it also reveal relatively high false positive rate. The concordance rate of test results between LG and Axsym was 96.2%, between LG and Cobas was 95.5%, and total agreement between three EIA kit was 93.9%. LG were relative poor distinction between positive and negative results, but it could be applied clinically as a screening tool for hepatitis C in general population. The SIC of one false negative result by LG was 0.91, and false positive were less than 4.0, therefore we concluded it is necessary to confirm by immunoblotting assay when SIC were between 0.8 and 4.0.

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Detection of Mutations to Zidovudine in the pol Gene of Human Immunodeficiency Virus-1 by Direct Sequencing (인면역결핍 바이러스 pol 유전자 염기서열 결정에 의한 지도부딘 (ZDV) 내성 돌연변이의 탐지)

  • Cho, Young-Keol;Lee, Hee-Jung;Sung, Heung-Sup;Kim, Yoo-Kyum;Kim, Young-Bong;Lee, Yong-Jin;Kim, Mi-Jung;Kim, Dae-Ghon;Won, Young-Ho;Cho, Goon-Jae
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.271-281
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    • 1999
  • The nested polymerase chain reaction (PCR) assay was used to determine the sequences of reverse transcriptase (RT) codons 41, 67, 70, 210, 215 and 219 of human immunodeficiency virus-1 (HIV-1) pol gene. Template DNA was obtained from uncultured peripheral blood mononuclear cells from 27 Korean HIV-1 infected patients treated with ZDV and Korean red ginseng. The second PCRs were done for 2 separated regions (RT codons $13{\sim}98$ and $152{\sim}259$) with $5\;{\mu}l$ of the first PCR productNucleotide sequences were determined by direct sequencing. In the 27 patients, CD4+ cell count decreased from $230{\pm}117/{\mu}l$ to $152{\pm}162/{\mu}l$ for $46{\pm}26$ months (Mo), and actual duration of ZDV intake was $72{\pm}16$ Mo. In the 16 patients who had been treated with ZDV therapy ${\ge}25$ Mo, the incidences of 70R, 215F/Y, and 41L were 61%, 28% and 22%, respectively and those of 67N, 210W and 219Q were 17%. The incidences of 215F/Y were 6.7% for group ${\le}12$ Mo treatment, 22.7% for group with 13 to 24 Mo, and 27.8% for group ${\ge}25$ Mo. There was no mutation in 9 patients. It might be associated with the interruption of ZDV therapy for more than 6 months in 6 patients. This study shows that the detection of mutation could be useful prognostic marker with other clinical and virological data, and very low mutation rate is dectected compared to overseas reports.

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Unstable Data of HBe Antigen during Seroconversion from HBe Antigen to Antibody in Chronic Type B Hepatitis (만성B형 간염에서 HBe 항원에서 항체로의 혈청 전환 중에 불안정하게 나타나는 HBe 항원)

  • Shin, Sun-Young;Min, Gyeong-Sun;Noh, Kyung-Woon;Kim, Hyun-Joo
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.1
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    • pp.78-81
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    • 2008
  • Purpose: In this study, we evaluated unstable serum data of HBe antigen (HBeAg) or HBe antibody (HBeAb) in patients who experienced HBeAg seroconversion. This study have been performed to assist a medical technologist in the recognition of patients who were chronically infected with the hepatitis B virus (HBV). Materials and Methods: A total number of 3 patients were enrolled in this study. All patients experienced HBeAg seroconversion. Serum data of HBeAg and HBeAb were measured by radioimmunoassay. Results: The data of HBeAg or HBeAb showed an unstable change during seroconversion from HBeAg to HBeAb in chronic type B hepatitis (CBH). Conclusions: Serum data of HBeAg or HBeAb can change during HBe seroconversion. These data suggest that patients with HBe seroconversion can experience an unstable oscillation of HBeAg or HBeAb value from positive to negative. Unstable data can appear naturally due to the seroconversion process.

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Epidemiological surveys of an outbreak of false positive classical swine fever in Gyeongbuk province (의사 돼지콜레라 발생농장 역학조사)

  • Park, No-Chan;Cho, Kwang-Hyun;Kim, Young-Hoan;Kim, Soon-Tae;Kim, Sung-Kuk;Park, In-Hwa;Cho, Min-Hee;Oh, Gang-Hee;Son, Jae-Kweon;Jyeong, Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.26 no.2
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    • pp.163-184
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    • 2003
  • This study was conducted to survey the farm which suffered from disease similar to classical swine fever(CSF) in Gyeongbuk province. Clinical signs appeared first in a few number of growing pigs which showed specific signs of diarrhea, depression, sleepiness, and reluctance to get up or to eat. Younger piglets may have appeared chilled, shiver and huddle together. As the disease progresses the affected pig's skin went red and purple. In histopathological signs, there were many haemorrhages throughout the body and larger haemorrhages in some organs such as lymph nodes. And there is a precipitous fall in the number of circulating leukocytes in the blood. In spite of insisting of farmer which did not vaccinate to classical swine fever, significant antibody production was detected in these affected pigs at enzyme-linked immuonsorbent assay. According to the above results at first glance, these affected pig suspected with CSF in clinical signs and histopathological lesions only. Because the symptoms and post-mortem picture were very similar to CSF, these false positive results would have been dangerous to diagnostician. But by reverse transcriptase polymerase chain reaction(RT-PCR) and comparative nucleotide sequence analysis, the disease was correctly diagnosed with post-weaning multisystemic wasting syndrome(PMWS) and porcine reproductive and respiratory syndrome(PRRS) compoundly. And the antigen which were detected the lesion similar to CSF virus, was confirmed with LOM vaccine strain of CSF. In most national CSF eradication program and in countries which are free of the CSF virus, vaccination against CSF is not practiced and generally is not allowed. But now in Korea, routine vaccination is practiced because of outbreaking the CSF repeatedly. When CSF is diagnosed the whole herd and other in contact animal are slaughtered continuously.

Production of Red-spotted Grouper Nervous Necrosis Virus (RGNNV) Capsid Protein Using Saccharomyces cerevisiae Surface Display (Saccharomyces cerevisiae 표면 발현을 이용한 붉바리 신경괴사 바이러스 외피단백질의 생산)

  • Park, Mirye;Suh, Sung-Suk;Hwang, Jinik;Kim, Donggiun;Park, Jongbum;Chung, Young-Jae;Lee, Taek-Kyun
    • Journal of Life Science
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    • v.24 no.9
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    • pp.995-1000
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    • 2014
  • The studies of marine viruses in terms of viral isolation and detection have been limited due to the high mutation rate and genetic diversity of marine viruses. Of the modern methods currently used to detect marine viruses, serological methods based on enzyme-linked immunosorbent assay (ELISA) are the most common. They depend largely on the quality of the antibodies and on highly purified suitable antigens. Recently, a new experimental system for using viral capsid protein as an antigen has been developed using the yeast surface display (YSD) technique. In the present study, the capsid protein gene of the red-spotted grouper nervous necrosis virus (RGNNV) was expressed and purified via YSD and HA-tagging systems, respectively. Two regions of the RGNNV capsid protein gene, RGNNV1 and RGNNV2, were individually synthesized and subcloned into a yeast expression vector, pCTCON. The expressions of each RGNNV capsid protein in the Saccharomyces cerevisiae strain EBY100 were indirectly detected by flow cytometry with fluorescently labeled antibodies, while recognizing the C-terminal c-myc tags encoded by the display vector. The expressed RGNNV capsid proteins were isolated from the yeast surface through the cleavage of the disulfide bond between the Aga1 and Aga2 proteins after ${\beta}$-mercaptoethanol treatment, and they were directly detected by Western blot using anti-HA antibody. These results indicated that YSD and HA-tagging systems could be applicable to the expressions and purification of recombinant RGNNV capsid proteins.