• Journal of Microbiology
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• v.38 no.3
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• pp.187-191
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• 2000

A validation study was conducted to determine the efficacy of solvent/Detergent (S/D) inactivation and Q-Sepharose column chromatographic removal of the human immunodeficiency virus (HIV) during the manufacturing of a high purity antihemopilic factor VIII (GreenMono) from human plasma. S/D treatment using the organic solvent, tri (n-butyl) phosphate, and the detergent, Trition X-100, was a robust and effective step in eliminating HIV-1. The HIV-1 titer was reduced from an initial titer of 8.3 log10 TCID50 to undetectable levels within one minute of S/D treatment, HIV-1 was effectively partitioned form factor VIII during Q-Sepharose column chromatography with the log reduction factor of 4.1 . These results strongly assure the safety of GreenMono From HIV.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2015.05a
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• pp.954-957
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• 2015

A recombinant baculovirus vector systems were composed of genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). These recombinant baculovirus vector system were transfected into various cell lines and tissues and confirmed gene transfer and expression of these vector systems with only control vector system. From the result, gene transfer and gene expression of recombinant baculovirus vector systems were superior in terms of efficacy and safety than in the control vector system.

• BMB Reports
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• v.53 no.11
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• pp.565-575
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• 2020

Gene therapy is emerging as a treatment option for inherited genetic diseases. The success of this treatment approach greatly depends upon gene delivery vectors. Researchers have attempted to harness the potential of viral vectors for gene therapy applications over many decades. Among the viral vectors available, gutless adenovirus (GLAd) has been recognized as one of the most promising vectors for in vivo gene delivery. GLAd is constructed by deleting all the viral genes from an adenovirus. Owing to this structural feature, the production of GLAd requires a helper that supplies viral proteins in trans. Conventionally, the helper is an adenovirus. Although the helper adenovirus efficiently provides helper functions, it remains as an unavoidable contaminant and also generates replication-competent adenovirus (RCA) during the production of GLAd. These two undesirable contaminants have raised safety concerns and hindered the clinical applications of GLAd. Recently, we developed helper virus-free gutless adenovirus (HF-GLAd), a new version of GLAd, which is produced by a helper plasmid instead of a helper adenovirus. Utilization of this helper plasmid eliminated the helper adenovirus and RCA contamination in the production of GLAd. HF-GLAd, devoid of helper adenovirus and RCA contaminants, will facilitate its clinical applications. In this review, we discuss the characteristics of adenoviruses, the evolution and production of adenoviral vectors, and the unique features of HF-GLAd as a new platform for gene therapy. Furthermore, we highlight the potential applications of HF-GLAd as a gene delivery vector for the treatment of various inherited genetic diseases.

• Journal of fish pathology
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• v.27 no.1
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• pp.11-16
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• 2014

The resistance against white spot syndrome virus (WSSV) infection in wild marine crab Gaetice depressus by the immunization of a recombinant glutathione-S-transferase (GST) fused VP28 protein (GST-VP28) was evaluated. The cumulative mortalities of GST-VP28 injected groups were lower than those of the control groups at 10 days of post-challenge, and the time to death of 50% crab ($TD_{50}$) was delayed by the immunization using GST-VP28. The group boosted with GST-VP28 after 2 weeks of primary immunization clearly showed longer $TD_{50}$ than non-boosted group against challenge with WSSV. This result suggests that boosting with the antigen protein elicit stronger immune responses similar to adaptive immune responses of vertebrates. However, the short $TD_{50}$ was observed in the group challenged at 3 weeks post boosting comparing to the group challenged at 1 week post boosting. This suggests that the protective strength of immunization decreased by the time.

• Journal of Life Science
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• v.22 no.6
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• pp.703-712
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• 2012

Adeno-associated virus (AAV) has been considered to be a very safe and efficient gene delivery system. However, the major obstacles to therapeutic usage of AAV have been to achieve highly efficient and reproducible production processes, and also to develop a reliable quantifying method of various serotypes with a simple protocol. We compared the efficiency of the conventional production protocol of AAV2 and adenovirus (Ad) co-infection to that of a new method containing AAV2 infection followed by pHelper transfection. We tested HEK293 and 293T, and further examined the time-dependent changes of AAV2 production. The new method of AAV2 and pHelper DNA gave about ten times higher production efficiency than that of the conventional protocol. The highest production efficiency in 293T was achieved as $1.61{\times}10^5$ virus genomes (v.g.)/cell by the new method of 10 MOI of AAV2 infection and 5 days post-infection. This protocol of the highest efficiency was then applied to produce various AAV serotypes and showed the efficiencies higher than $10^5$ v.g./cell. Next, we designed the universal PCR primers of highly conserved regions for various AAV serotypes to develop a simple and reliable titration method. The universal primers could amplify all the tested AAV serotypes with similar sensitivities by ten molecular copies. Therefore, this pair of universal primers can be further utilized to detect AAV contaminants in therapeutic adenoviral vectors.

• Journal of the Korean Society of Safety
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• v.22 no.4
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• pp.102-109
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• 2007

A bioterrorism attack is the deliberate release of viruses, bacteria, or other germs(agents) used to cause illness or death in people, animals, or plant. These agents are found in nature, but it is possible that they could be changed to increase their ability to cause disease, make them resistant to current medicines, or to increase their ability to be spread into the environment. Terrorists may use biological agents because these agents can be extremely difficult to detect and do not cause illness for several days. Some bioterrorism agents, like smallpox virus, can spread from person to person, like anthrax, can not. From these agents, we discussed the characteristics of biological agents and national safety regulation on the weapons of mass destruction including bioterrorism.

• Journal of Food Hygiene and Safety
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• v.29 no.1
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• pp.31-39
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• 2014

Norovirus causes acute gastroenteritis in all age groups and its food poisoning outbreaks are rapidly increasing in Korea. Reverse transcription-polymerase chain reaction (RT-PCR) is most widely used for the rapid detection of foodborne viruses due to high sensitivity. However, the false positive results of RT-PCR obtained against already inactivated viruses could be a serious drawbacks in food safety area. In this study, we investigated a method to yield true positive RT-PCR results only with alive viruses. To decompose the RNA genes from dead viruses, the enzymatic treatments composed of proteinse K and Ribonuclease A were applied to the sanitized and inactivated virus particles. Another aim of this study was to quantify the efficiencies of several major sanitizing treatments using real-time RT-PCR. Feline calicivirus (FCV) that belongs to the same Caliciviridae family with norovirus was used as a surrogate model for norovirus. The initial level of virus in control suspension was approximately $10^4$ PFU/mL. Most of inactivated viruses treated with the enzymatic treatment for 30 min at $37^{\circ}C$ were not detected in RT-PCR, Quantification results to verify the inactivation efficiencies of sanitizing treatments using real-time RT-PCR showed no false positive in most cases. We could successfully develope a numerical quantification process for the inactivated viruses after major sanitizing treatments using real-time RT-PCR. The results obtained in this study could provide a novel basis of rapid virus quantification in food safety area.

• Korean Journal of Veterinary Service
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• v.45 no.2
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• pp.117-124
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• 2022

In the event of an outbreak of a livestock epidemic, it has been considered that the existing burial-centered carcass disposal method should be improved ecofriendly for prevention of leachate and odors from burial basically in regard of pathogen inactivation. Therefore, the aim of this study is whether it was possible to treat the carcass of cattle and chickens using the chemical carcass treatment method. It was conducted to establish detailed treatment standards for the chemical treatment method of cattle and chicken carcasses based on the results of the proof of the absence of infectious diseases in cattle chickens. After inoculating cattle carcass with 10 pathogens (foot and mouth disease virus, bovine viral diarrhea virus, Mycobacterium bovis, Mycobacterium avium subsp. Paratuberculosis, Brucella abortus, Bacillus anthracis, Clostridium chauvoei, Clostridium perfringens, Escherichia coli, and Salmonella Typhimurium) and chicken carcasses with low pathogenic avian influenza virus, Clostridium perfringens type C, E. coli and Salmonella Typhimurium, these were treated at 90℃ for 5 hours in a potassium hydroxide liquid solution corresponding to 15% of the body weight. This method liquefies all cadaveric components and inactivates all inoculated pathogens by PCR and culture. Based on these results, it was possible to prove that chemical treatment of cattle and chicken carcasses is effective in killing pathogens and is a safe method without the risk of disease transmission. The chemical treatment method of livestock carcasses can be suggested as an alternative to the current domestic burial-centered livestock carcass treatment method, preventing environmental pollution, and contributing to public health.

• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.255-261
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• 2012

Foot and mouth disease (FMD) is highly infectious disease of cloven-hoofed animals, particularly cattle, sheep, pigs and goats. Last outbreak reported in November, 2010 induced the enormous social and economical impacts. Culling of infected animals, movement control, and vaccination are the major control measures of FMD. The aim of this study was to detection foot-and-mouth disease virus (FMDV) in the soil and leachate from modeling burial for pig carcass as measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). FMDV and Coxsakievirus B1 (CVB1) were detected in soil by week 16 and Coxsakievirus B1 (CVB1) by weeks 12, respectively. FMDV and CVB1 also detected by weeks 8 in the leachate. Results from this study provides an evidence that FMDV could be inactivated for safe of pig carcasses infected with FMDV within 4 month in the carcass burial site.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.321-327
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• 2007

This is a case report on the occurrence of inclusion body hepatitis (IBH) and infectious bursal disease (IBD) among the broilers in a local farm located in Wanju, Jeollabukdo. Mostly IBH could be caused by adenovirus if the bird's immune system was first weakened by exposure to immunosupressive agents such as infectious bursal disease virus (IBDV) and chicken anemia virus (CIAV). However IBH primary occurred before IBD in this case. And recent work has demonstrated that virulent adenovirus alone can produce the disease.