• Korean Journal of Microbiology
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• v.35 no.1
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• pp.28-34
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• 1999

Fragmentation vectors are used to analyze function and genomic structure of a gene of interest by creating deletion derivatives of large fragments of genomic DNA cloned as yeast artificial chromosomes (YACs). Herein, we developed a new hicistronic fragmentation vector that contains internal ribosomal entry sile (IRES) of encephalomyocarditis vin~s (EMCV) and $\beta$-galactosidase as a reporter gene. This vector system provides a novcl loo1 to analyze expression patterns of a gene of interest due to simultaneous expression of a target gene as well as $\beta$-galactosidase driven from a single message. In addition, the bicistronic fragmentation vector contains four rare-cutting restriction enzyme sites in the polycloning sites which can be used to conveniently insert any kinds of genes and therefore facilitates targeting DNA scgments into YAC by means of homologous recombination. This approach establishes a paradigm for manipulation of mammalian DNA segments and characterization of expression and regulatory regions of mammalian gene cloned as YAC.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.19-25
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• 2006

With particular regards to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor IX concentrate. The loss of factor IX activity during dry-heat treatment was of about 3%, as estimated by a clotting assay. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor IX compared with those of the factor IX before dry-heat treatment. The dry-heat-treated factor IX was stable for up to 24 months at $4^{\circ}C$, The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV and murine encephalomyocarditis virus (EMCV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Porcine parvovirus (PPV) and bovine herpes virus (BHV) were potentially sensitive to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\ge}5.60$ for HAV, ${\ge}6.08$ for EMCV, 2.64 for PPV, and 3.59 for BHV. These results indicate that dry-heat treatment improves the virus safety of factor IX concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.997-1003
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• 2008

Viral safety is a prerequisite for manufacturing clinical antihemophilic factor VIII concentrates from human plasma. With particular regard to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor VIII concentrate. The loss of factor VIII activity during dry-heat treatment was of about 5%. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor VIII compared with those of the factor VIII before dry-heat treatment. The dry-heat-treated factor VIII was stable for up to 24 months at $4^{\circ}C$. The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV, murine encephalomyocarditis virus (EMCV), and human immunodeficiency virus (HIV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Bovine herpes virus (BHV) and bovine viral diarrhea virus (BVDV) were potentially sensitive to the treatment. However porcine parvovirus (PPV) was slightly resistant to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\geq}5.55$ for HAV, ${\geq}5.87$ for EMCV, ${\geq}5.15$ for HIV, 6.13 for BHV, 4.46 for BVDV, and 1.90 for PPV. These results indicate that dry-heat treatment improves the virus safety of factor VIII concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid-enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

• YAKHAK HOEJI
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• v.50 no.2
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• pp.65-69
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• 2006

Novel 4'-hydroxymethyl branched thioapiosyl nucleosides were synthesized in this study. The introduction of hydroxymethyl group in the 4'-position was accomplished by a [3,3]-sigmatropic rearrangement. Thioapiosyl sugar moiety was constructed by sequential ozonolysis, reduction and cyclization. The pyrimidine nucleosidic bases (uracil, 5-fluorouracil, 5-iodouracil, 5-chlorouracil, 5-bromouracil) were efficiently coupled by Vorbruggen glycosyl condensation procedure (per-silyated base and TMSOTf). The antiviral activities of the synthesised compounds were evaluated against the HIV-1, HSV-1, HSV-2 and EMCV 5-Iodouracil 18 showed weak antiviral activity against HSV-1 $(EC_{50}=30.7{\mu}M)$.

• Korean Journal of Pharmacognosy
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• v.30 no.1
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• pp.25-33
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• 1999

In order to find less toxic antiviral agents from basidiomycetes, EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata (Pers.) Karst. EA was examined for antiviral activity against five strains of pathogenic viruses such as encephalomyocarditis virus (EMCV), vesicular stomatitis virus (VSV) Indiana and New Jersey strains, influenza A virus (Flu A), and varicella zoster virus (VZV) in vitro. Antiviral activity was evaluated by plaque reduction assay. Among five strains of viruses tested, EA exhibited the most potent antiviral activity against VSV Indiana strain with 50% effective concentration $(EC_{50})$ of 0.104 mg/ml in Vero cells, and its selectivity index (SI) was 36.5. EA was also examined for the virucidal activity, antiviral activity in preincubation on VSV Indiana strain in order to examine possible mode of antiviral activity. Preincubation of Vero cells with EA did not confer protection against VSV, however, prolonged exposure of cells to EA inhibited the replication of virus dose-dependently. In virucidal activity, the titer of infectious virus did not decrease significantly.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.765-772
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• 1997

제주지역 돼지에서 각종 전염성 질병 원인체에 대한 항체를 조사하여 그간 전염성 병원체에 대한 역학조사가 미진하였던 부분을 보완하여 질병의 분포를 파악하고자 1995년부터 1996년에 걸쳐 제주도 전역에서 돼지의 혈청을 채취하여 각종 병원체에 대한 항체 분포율을 조사하였다. 본 연구에서 검사한 돼지 혈청 시료에서는 돼지 오제스키병 바이러스에 대한 항체는 전혀 검출되지 않았다. 돼지 콜레라바이러스에 대한 항체는 기대 수준 이하로 낮아 백신접종이 원활히 수행되고 있지 않음을 시사하였으며 특히 농장에 따라 항체 보유돈과 항체 음성돈이 혼재하는 농장과 항체가 전혀 검출되지 않는 농장 등 돼지 콜레라 방역의 사각지대가 존재할 가능성이 있음을 보여주었다. 유 사산 원인체인 돼지 파보바이러스 및 뇌심근염에 대한 항체가가 다양하게 나타나 일부 문제가 있을 것으로 사료되었다. 돼지 생식기호흡기증후군(PRRS) 바이러스에 대한 항체 분포율은 내륙 보다 다소 낮게 나타났고, 돼지 influenza virus, 위축성 비염, 흉막 폐염 등 각종 세균성 질환에 대한 항체수준도 다양하게 나타났다. 본 혈청학적인 연구결과는 제주지역에서의 양돈방역 정책수립 및 질병방제의 기초자료로 유용하게 이용될 것으로 사료된다.

• Korean Journal of Veterinary Service
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• v.34 no.2
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• pp.111-116
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• 2011

Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.77-83
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• 1994

Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.