• Title/Summary/Keyword: Virus culture

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Epidemics of Lower Respiratory Tract Infection that Occurred to Children at a Local Institute for Children's Adoption and Welfare (모 입양 기관에 수용 중인 영유아에서의 하기도 감염의 유행)

  • Yang, Mi Hwa;Park, Chan Sook;Yoon, So Young;Kim, Jae Youn;Ahn, Don Hee
    • Pediatric Infection and Vaccine
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    • v.9 no.2
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    • pp.201-207
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    • 2002
  • Purpose : We analyzed clinical features and causal viruses of acute lower respiratory tract infections(LRTIs) in order to improve the management of these infections. Methods : From end of April to early May 2001, amongst 30 children at a local institute for children's adoption and welfare, 13 were admitted to the hospital with the diagnostic impression of acute LRTIs. Nasopharyngeal aspirates were sent in Seoul National University Hospital for viral culture of respiratory syncytial virus(RSV), adenovirus, parainfluenza virus. Results : One or more viral agents were identified in 4 cases(30.7%) : were RSV(15.4%), adenovirus(7.7%), and a mix of these two viruses(7.7%). Initial symptoms were fever(69%), cough(100%), tachypnea(54%), chest retraction(69%), rale(85%) and wheezing(15%). Leukocytosis was noted in 23%, CRP increased more than 10 mg/L in 46%. Chest X-ray abnormalities were 69%. Conclusion : Although viruses were identified in 30.7%, further studies should be made for prevention and treatment of acute viral LRTIs.

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The inhibition effects of Injinchunggantang on Hepatitis B and C Virus (인진청간탕(茵蔯淸肝湯)의 B형(型) 및 C형(型) 간염 바이러스에 대한 항(抗) 바이러스 효과(效果))

  • Kim, Su-Sung;Kim, Gi-Yeol;Yoon, CheoI-Ho;Seo, Woon-Gyo;Kim, Jong-Dae;Chung, Ji-Choen
    • The Journal of Dong Guk Oriental Medicine
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    • v.8 no.1
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    • pp.93-106
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    • 1999
  • For the purpose of investigate the inhibition effects of Injinchunggantang on Hepatitis and C Virus. The inhibition experiments were as follows : 1. The inhibition effects of Hepatitis B surface antigen production was remarkably increased in the pretreated group of Injinchunggantang compared with control group. 2. Amplified PCR products of HBV-DNA in culture media was considerably decreased in the pretreated group of Injinchunggantang compared with control group. 3. Results of ATP quantitation and ATPase inhibition percent was increased pretreated group of Injinchunggantang. Also ATPase activity of HCV helicase inhibition assay was increased with a dose dependent manner. These results may suggest that Injinchunggantang will have the inhibition effects of Hepatitis B and C Virus.

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Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.147-152
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    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

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Application of chemical treatment for cattle and chicken carcasses for the control of livestock infectious diseases (가축 전염병 발생에 따른 소와 닭 사체의 화학적 처리 방법의 적용)

  • Lee, Taek Geun;Oh, Yeonsu;Ko, Young-Seung;Bae, Da-Yun;Tark, Dong-Seob;Rim, Chaekwang;Cho, Ho-Seong
    • 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.

The Protein and Isozyme Patterns During in vitro Plant Regeneration of Yooja (Citrus junos Sieb.) and Trifoliate Orange (Poncirus trifoliata Rafin.)

  • Park, Min-Hee;Jang, Hyun-Kyu;Cha, Young-Ju;Kim, Ho-Bun;Lee, Sook-Young
    • Plant Resources
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    • v.5 no.1
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    • pp.29-44
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    • 2002
  • In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

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Real-Time AT-PCR for Quantitative Detection of Bovine Parainfluenza Virus Type 3 during the Manufacture of Biologics (생물의약품 제조공정에서 Bovine Parainfluenza Virus Type 3 정량 검출을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Kim, Chan-Kyong;Kim, Tae-Eun;Kim, In-Seop
    • KSBB Journal
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    • v.23 no.4
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    • pp.303-310
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    • 2008
  • Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biologics such as biopharmaceuticals, tissue-engineered products, and cell therapy. Manufacturing processes for the biologics have the risk of viral contamination. Therefore viral validation is essential in ensuring the safety of the products. Bovine parainfluenza virus type 3 (BPIV3) is one of the common bovine pathogens and has widely been known as a contaminant of biologics. In order to establish the validation system for the BPIV3 safety of biologics, a real-time RT-PCR method was developed for quantitative detection of BPIV3 contamination in raw materials, manufacturing processes, and final products. Specific primers for amplification of BPIV3 RNA was selected, and BPIV3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 2.8 $TCID_{50}/mL$. The real-time RT-PCR method was validated to be reproducible and very specific to BPIV3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BPIV3. BPIV3 RNA could be quantified in CHO cell as well as culture supernatant. Also the real-time RT-PCR assay could detect 7.8 $TCID_{50}/mL$ of BPIV3 artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BPIV3 contamination during the manufacture of biologics.

Yearly Occurrence of Thrips Infesting Hot Pepper in Greenhouses and Differential Damages of Dominant Thrips (시설 고추재배지 총채벌레 연중 발생 및 주요 총채벌레의 차등 해충성)

  • Kim, Chulyoung;Choi, Duyeol;Lee, Donghyun;Khan, Falguni;Kwon, Gimyon;Ham, Eunhye;Park, Jungjoon;Kil, Eui-Joon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.61 no.2
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    • pp.319-330
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    • 2022
  • Andong is a place to culture the great amount of hot pepper (Capsicum annuum L.) in Korea. This study reports a yearly occurrence (March 31~October 25, 2021) of thrips infesting the hot pepper in Andong. Thrips caught to yellow sticky traps were diagnosed by morphological characters and showed two dominant species: Frankliniella occidentalis and F. intonsa. During this period, a total of 107,874 thrips were caught and included F. occidentalis at about 82%, F. intonsa at about 17%, and the other thrips at about 0.3%. There were two main peaks at May~June and at September~October, respectively, in which the total number of thrips was higher in the second peak and most were F. occidentalis. Interestinly, a low level of thrips occurred during July~August was observed and explained by their susceptibility to high temperatures. A laboratory experiment by exposing thrips to high temperatures showed that thrips were susceptible to temperatures higher than 35℃ and not tolerant to 45℃ for 1 h. Indeed, high temperatures higher than 45℃ were recorded in the greenhouses in Andong during July~August. F. occidentalis was more tolerant to the high temperatures than F. intonsa. On the other hand, the thrips showed the highest occurrence peak at July~August in hot pepper-culturing greenhouse in Kangwon, where the average temperatures were mostly lower than those of Andong and no high temperatures higher than 45℃ were recorded during July~August. A viral disease caused by tomato spotted wilt virus (TSWV) was observed in the hot peppers cultured in Andong greenhouses. Multiplex PCR was used to detect the virus along with identification of thrips. With a high record of about 30%, the virus-infected thrips were detected during all the monitoring period. The virulent thrips were identified to be only F. intonsa. These results suggest that F. occidentalis gives a direct damage especially during harvesting period with their high populations while F. intonsa gives indirect damage by transmitting TSWV.

Evaluation of stability and potency of live attenuated rinderpest vaccine of lapinized-avianized tissue culture strain origin for the establishment of expiration period (가토화·계태화 우역 생백신의 유효기간 설정을 위한 안정성 및 효능 평가)

  • Yeh, Jung-Yong;Kim, Hyun-Ju;Nah, Jin-Ju;Park, Jee-Yong;Lee, Ji-Hye;Seo, Hyun-Ji;Kweon, Chang-Hee;Cho, In-Soo;Moon, Jin-San
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.201-205
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    • 2009
  • In this study, we evaluated the stability and potency of live attenuated rinderpest vaccines of lapinized-avianized tissue culture strain origin, which had been produced annually from 2005 to 2008. When immune responses to the vaccines were evaluated using two Holstein calves weighing 100~150 kg, neutralizing antibody titer of 1 : 16 was induced at 21 days post vaccination. When calves were also inoculated with vaccines lots that had been stored for 39 months at ${4^{\circ}C}$, same level of antibody titer was observed. Using the virus titer test, we found that all batches of the vaccine that had been kept for 3, 10, 15, 22, 27, 34, 39, and 45 months showed no significant loss of titers, and fulfilled the requirement necessary ($\geq$ 3 $logTCID_50$) to be used as the national rinderpest vaccine reserve in Korea. In this study, we demonstrated that stability and potency of the rinderpest vaccines were maintained over three years when kept at ${4^{\circ}C}$ storage. This indicates that it maybe feasible to extend the expiration period of this vaccine from one year to three years.

Effects of Insect Hormones on the Replication of Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Yi, Yong-Zhu;Xiao, Qing-Li;He, Jia-Lu;Zhou, Ya-Jing;Zhang, Yuan-Xing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.2
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    • pp.137-141
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    • 2002
  • An experimental study was undertaken to quantify the effects of insect hormones on the replication of nucleopolyhedrovirus (NPV). The results demonstrated that TCID/ sub 50/ at 72 h post-infection (hpi) rose systematically from 0.55$\times$10$^{8}$ /m1, for untreated cells, up to 1.67$\times$10$^{8}$ / ml at 3$\mu$g/ml, then dropped down to 1.45$\times$10$^{8}$ /m1 at 4 $\mu$g/ml, by adding ecdysone to the culture medium for Bm-N cells infected with a wild-type Bambyx mori. nucleopolyhedrovirus (BmNPV). The optimum enhancement of about 3 times on budded virus (BV) titer at 72 hpi was given at 3 $\mu$g/ml of ecdysone. While the polyhedra number had no obvious variation within the range of concentrations from 0 to 4 $\mu$g/ml. By addition of juvenile hormone analogue (JHA) into the media with this concentration range, the BmNPV TCID/ sub 50/ and polyhedra number at 72 hpi did not show significant changes. Also, the addition of either 3 $\mu$g/ml of ecdysone or 3 $\mu$g/ml of JHA to the culture media did not appear to affect the TCID/ sub 50/ and polyhedra number significantly in infected Sf-21 cells with the autographa californica nucleopolyhedrovirus (AcMNPV).

Establishment of Immotalized Human Gingival Fibroblast Cell Lines (불멸화된 치은 섬유아 세포주의 확립)

  • Song, Jae-Bong;Kim, Hyun-A;Hyun, Ha-Na;Kim, Eun-Cheol;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.603-614
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    • 2002
  • Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamHl and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.