• Title/Summary/Keyword: VERO

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Growth Characteristics of an Attenuated Japanese Encephalitis Virus in a Monkey Kidney Cell (Vero) (베로 세포에 적응된 약독화 일본뇌염바이러스의 성장 특성)

  • 홍선표;정용주;문상범;신영철;이성희;김수옥
    • KSBB Journal
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    • v.13 no.3
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    • pp.231-237
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    • 1998
  • An attenuated Japanese encephalitis virus (JEV) clone SA-14-14-2(Vero) was obtained through successive adaptation of a primary cell adapted strain, SA-14-14-2(PDK) to Vero cell, a continuous line of monkey kidney cells. The virus titer reached above the 107 plaque forming unit (pfu) per mL of culture supernatant with 3 passages in Vero cells and was maintained close to this level in the further passages. The optimum temperature for the virus growth was $35^{\circ}C$. Growth pattern of the virus indicated that optimum time for the virus harvest is 4 days post infection and the virus accomplished rapid initial propagation even in medium containing no serum supplement. The roller bottle (RB) system and the spinner flask (SF) system using micro-carrier (Cytodex-1) for the JEV cultivation were explored. When RB, SF, and T-flask culture system were compared, there was no significant difference in virus yield. Furthermore, the results indicated that virus could be harvested multiple times from 3 days to 9 days post infection; neither severe cytopathic effect (CPE) in the infected cells nor the decrease in the titer was observed on duration of 9 days.

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The Effects of Vero Cells Coculturing on the Motility of Human Testicular Spermatozoa in an Intracytoplasmic Sperm Injection Program (정자직접주입술에 있어 Vero Cells 공배양이 인간 정소정자의 운동성에 미치는 영향에 관한 연구)

  • Kim, Hyun-Kyoo;Oum, Ki-Boong;Kim, Hyun-Joo;Ko, Jung-Jae;Lee, Sook-Hwan;Yoon, Tae-Ki;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.225-232
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    • 1997
  • 폐색성 혹은 비폐색성 무정자증에서 부정소 정자채취법 등이 부적절하다고 여겨질때는 정소 조직을 일부 절제하여 그 조직으로부터 정자를 직접 채취하게 되는데 일반적으로 이렇게 정소로부터 추출한 정소정자는 운동성이 전혀 없거나 매우 약한 운동성을 보이는 경우가 많다. 본 연구의 목적은 이러한 정소정자를 Vero cell과 공배양을 시킴으로써 운동성을 획득시키거나 향상시키고 이를 수정시키는 시기까지 지속시킴으로써 정소정자추출술 (TESE)을 시행하는 환자나 의료진들에게 보다 편안하고 융통성있는 시간대를 부여하고, 아울러 정자직접주입술 (ICSI)을 보다 용이하게 하여 성공적인 수정률과 임신율을 얻음에 있다. 또한 ICSI를 시행한 후, 운동성이 향상된 잉여의 정소정자를 냉동보존함으로써 차후에 TESE을 다시 시행치않고도 시험관 아기 시술을 시도할 수 있는 부가적인 잇점도 있다고 할 수 있다. 대상환자군은 정관폐색증(n=11) 혹은 비정관폐색증(n=2)을 보이는 13명의 무정자증의 남성불임환자였으며 난자회수예정일 3일전에 TESE를 시행하여 정소정자를 얻은 후 이를 정자직접주입술이 시행되는 당일까지 Vero cell과 공배양을 실시하였다. Vero cell과의 공배양에 의하여 운동성이 있는 정소정자의 수는 공배양전과 비교하여 평균 3.3배가 증가하였으며, 특히 공배양전에 운동성이 있는 정소정자의 수가 50,000/ml이하의 미약한 운동성만을 보였던 경우 (n=5)에는 공배양 후에 운동성이 있는 정소정자 수의 평균증가율이 7.7배였다. 공배양전 정자운동성이 전혀 없었던 2례의 비정관폐색증환자중 3일간의 공배양을 통하여 1례에서 운동성을 획득한 정소정자를 얻을 수 있었으며 (14,300/ml), 정자직접주입술을 통하여 성공적인 수정 및 임신에 도달할 수 있었다. Vero cell과 공배양을 하고 ICSI했던 결과, 평균 수정률은 75.0% 이었으며 임신율은 61.5%였다.

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Isolation and molecular characterizations of canine distemper virus from a naturally infected Korean dog using Vero cells expressing dog signaling lymphocyte activation molecule

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Yoon, Yoon-Seek;Park, Jungwon;Oh, Dongryul;Yoo, Jae Young;Ji, Miryeon;Han, Bokhee;Oh, Subin;Hyun, Bang-Hun
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.64.1-64.14
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    • 2020
  • Background: Canine distemper virus (CDV) infection results in high morbidity and mortality in dogs. There has been no report about Isolation of Korean CDV since 1980 in Korea. Objectives: To investigate the biological properties and the genetic characterization of Korean CDV. Methods: Vero cells expressing dog signaling lymphocyte activation molecule (dSLAM) gene named as Vero/dSLAM were used to isolate CDV using 17 samples. Diagnostic methods such as cytopathic effects, immunofluorescence assay, peroxidase linked assay, electron microscopy, rapid immunodiagnostic assay, and reverse transcription polymerase chain reaction were used to confirm the Korean CDV isolate as a CDV. The genetic analysis was performed through cloning and sequencing of hemagglutinin gene of CDV isolate. Results: A virus propagated in Vero/dSLAM cell was confirmed as CDV (CD1901 strain) based on the above methods. The CD1901 strain showed the highest viral titer (105.5 50% tissue culture infectious dose [TCID50]/mL) in the Vero/dSLAM cells at 4 days post inoculation, but did not form a fork on chorioallantoic membrane of 7-day-old egg. Ribavirin, a nucleotide analogue anti-viral agent, inhibits moderately the Korean CDV propagation in the Vero/dSLAM cells. The nucleotide and amino acid sequences of the H gene of CD1901 strain were compared with those of other CDV strains. The CD1901 strain belonged to Asia 1 group and had the highest similarity (99.9%) with the BA134 strain, which was isolated in China in 2008. Conclusions: We constructed successfully Vero/dSLAM and isolated one Korean CDV isolate (CD1901 strain) from a naturally infected dog. The CD1901 strain belonged to Asia 1 genotype.

The Inhibitory Effects of Quercetin from Houttuynia cordata against Cadmium-Induced Cytotoxicity (어성초로부터 분리된 Quercetin의 카드뮴에 대한 독성억제효과)

  • Choi, Hwa-Jung;No, Yong-Ju;Kang, Jeong-Il;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.53 no.1
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    • pp.12-18
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    • 2009
  • The aim of this study was to develop antitoxic compound for cadmium-induced cytotoxic Vero cells. These cells were divided into five groups; control group (medium only), cadmium group (cadmium only), and two experimental groups. SRB (sulphorodamine B) assay was performed to evaluate the cytotoxicity of cell organelles. After cadmium was treated on Vero cells, we determined IC50 values to examine the detoxification effects of Houttuynia cordata methanol extract and quercetin under cadmium-induced cytotoxicity. Furthermore, morphological changes were observed by the light microscope. In Vero cells, methanol extract of Houttuynia cordata, and quercetin showed inhibitory effects on cadmium-induced cytotoxicity and these detoxification effects were increased in a concentration-dependent manner. These results suggest that methanol extract and quercetin from Houttuynia cordata retain a potential antitoxic activity.

The Effect of Co-culture and Oxygen Concentration on In Virto Fertilization of Follicular Oocytes in Korean Native Cattle (공배양 및 산소농도가 한우 난포란의 체외발생에 미치는 영향)

  • 이재관;윤준진;황성수;윤종택;김창근;정영채
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.43-50
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    • 1998
  • The effect of oxygen tension on embryonic development in co-culture was evaluated from the standpoint of the reduction of dissolved oxygen concentration by the oxygen consumption of feeder cells. Three co-culture systems using bovine oviductal epitherial cells (BOEC), African green monkey kidney cells (Vero cells) or buffalo rat liver cells (BRLC) have been compared in terms of development of bovine embryos derived from oocytes matured and fertilized in vitro. Among the co-cultured embryo, Vero cells su, pp.rted the highest developmental rate (29%) and the other two showed the similar rates. When the co-cultures were incubated in three different oxygen tension such as 5, 10, 20% oxygen atmosphere, embryos co-cultured with Vero cells at 10%-O2 resulted in the highest percentage of development. From the measurement of oxygen consumption of feeder cells, BRLC consumed 1.38 10-10 mg-O2/min/cell which was higher than 0.94 10-10 and 0.26 10-10mg-O2/min/cell for Vero cells and BOEC, respectively. Based on the oxygen consumption data, the phenomena of optimum oxygen tension required in embryo development in vitro has been analyzed, and we suggested that gas phase oxygen concentration, oxygen consumption rate of feeder cells and the number of feeder cells should be considered for the design of optimal co-culture system for effective fertilization of embryos in vitro.

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Comparative susceptibility of different cell lines for culture of Toxoplasma gondii in vitro (톡소플라스마 곤디의 세포내 배양에 있어서 세포 주에 따른 감수성 비교)

  • 박병규;문형로
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.215-222
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    • 1993
  • In order to establish a useful cell culture system for T gondii we compared the degree of proliferation of T gondii tachyzoites among 8 different cell lines: 2 kinds of normal animal cells (MDCK-canine kidney cells; Vero-monkey kidney cells) and 6 kinds of human tumor cells (A 549, PC 14-lung cancer cells; SNU 1, SNU 16. Mlm 45-stomach cancer cells; HL-60-promyelocytic leukemia cells), through morphological observation and 3H-uracil uptake assay. The degree of susceptibility to infection with T gondii tachyzoites was highest in A 549 and PC 14 cells, medium in Vero, HL-60, MDCK and SNU 1, and lowest in SNU 16 and MBm 45 cells. The kinetics of T gondii multiplication during the post-Infection 60 hours were higllly dependent upon the dose of tachyzoites administered and the duration among the 8 tested fur the growth and multiplication of T gondii in vitro.

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Assessment on Accuracy of Stereotactic Body Radiation therapy (SBRT) using VERO (VERO system을 이용한 정위적 체부 방사선치료(SBRT)의 정확성 평가)

  • Lee, Wi Yong;Kim, Hyun Jin;Yun, Na Ri;Hong, Hyo Ji;Kim, Hong Il;Baek, Seung Wan
    • The Journal of Korean Society for Radiation Therapy
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    • v.31 no.1
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    • pp.17-24
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    • 2019
  • Purpose: The present study aims to assess the level of coherency and the accuracy of Point dose of the Isocenter of VERO, a linear accelerator developed for the purpose of the Stereotactic Body Radiation Therapy(SBRT). Materials and Method: The study was conducted randomly with 10 treatment plans among SBRT patients in Kyungpook National University Chilgok Hospital, using VERO, a linear accelerator between June and December, 2018. In order to assess the equipment's power stability level, we measured the output constancy by using PTW-LinaCheck, an output detector. We also attempted to measure the level of accuracy of the equipment's Laser, kV(Kilo Voltage) imaging System, and MV(Mega Voltage) Beam by using Tofu Phantom(BrainLab, Germany) to assess the accuracy level of geometrical Isocenter. We conducted a comparative analysis to assess the accuracy level of the dose by using an acrylic Phantom($30{\times}30{\times}20cm$), a calibrated ion chamber CC-01(IBA Dosimetry), and an Electrometer(IBA, Dosimetry). Results: The output uniformity of VERO was calculated to be 0.66 %. As for geometrical Isocenter accuracy, we analyzed the error values of ball Isocenter of inner Phantom, and the results showed a maximum of 0.4 mm, a minimum of 0.0 mm, and an average of 0.28 mm on X-axis, and a maximum of -0.4 mm, a minimum of 0.0 mm, and an average of -0.24 mm on Y-axis. A comparison and evaluation of the treatment plan dose with the actual measured dose resulted in a maximum of 0.97 % and a minimum of 0.08 %. Conclusion: The equipment's average output dose was calculated to be 0.66 %, meeting the ${\pm}3%$ tolerance, which was considered as a much uniform fashion. As for the accuracy assessment of the geometric Isocenter, the results met the recommended criteria of ${\pm}1mm$ tolerance, affirming a high level of reproducibility of the patient's posture. The difference between the treatment plan dose and the actual measurement dose was calculated to be 0.52 % on average, significantly less than the 3 % tolerance, confirming that it obtained predicted does. The current study suggested that VERO equipment is suitable for SBRT, and would result in notable therapeutic effect.

Prophylactic Effects of Bifidobacterium longum HY8001 against Escherichia coli O157:H7 and Salmonella typhimurium DT104 Enteric Infection and Evaluation of Vero Cytotxin Neutralizing Effects (Bifidobacterium longum HY8001 균주의 Escherichia coli O157:H7과 Salmonella typhimurium DT104 장곤 내 감염 예방효과 및 Vero cytotoxin 중화효과)

  • 양수진;윤장원;서근석;구혜정;김소현;배형석;백영진;박용호
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.419-425
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    • 1999
  • Prophylactic effects of Bifidobacterium longum HY8001, Korean isolate, against Escherichia coli O157:H7 and Salmonella typhimurium DT104 enteric infection were examined at four groups of specific pathogen free(SPF)-ICR mouse for each pathogen. B. longum HY8001+B. typhimurium DT104+B. longum HY8001(BL+ST+BL) group and B. longum HY8001+E. coli O157:H7+B. longum HY8001(BL+E+BL) group were fed with B. longum HY8001 before and after E. coli O157:H7 or s. typhimurium DT104 challenge, while B. longum HY8001+S. typhimurium DT104(BL+ST) and B. longum HY8001+e. coli O157:H7(BL+E) groups were fed with B. longum HY8001 only before E. coli O157:H7 or S. typhimurium DT104 challenge. E. coli O157:H7(E) and S. typhimurium DT104(ST) groups were challenged with each pathogen without B. longum HY8001 administration and control groups were administered with phosphate buffered solution(PBS). After the oral administration with B. longum HY8001(109cfu), th emice were challenged with E. coli O157:H7(2$\times$1010cfu) or S. typhimurium DT104(108cfu) and the mortality rate and the fecal shedding of challenged pathogen were also examined define the reactivity of the B. longum HY8001. Production of toxin neutralizing substance(s) of B. longum HY8001 was determined by cell cytotoxicity assay using Vero cells. Fecal shedding of th eS. typhimurium DT104 was significantly decreased in BL+ST+BL group fed with B. longum HY8--1 before and after challenge(p<0.05), while the fecal shedding s of S. typhimurium DT104 in BL+ST and St groups remained more than 106cfu. the protective effect of the B. longum HY8001 against E. coli O157:H7 was significantly high only in BL+E+BL group fed with b. longum Hy8001 before and after E. coli O157:H7 challenge from the result of fecal E. coli O157:H7 isolation rate, mortality rate, and intestinal contents culture to detect E. coli O157:H7. the mortality rate of the BL+e and E groups. The cytopathic effect (CPE) of the Vero cytotoxin (Shiga like toxin I & II) in Vero cell was neutralized in B. longum HY8001 culture supernatant added wells which indicate the presence of soluble Vero cytotxin neutralizing substance(s) in B. longum HY8001 culture suprnatant.

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Cytotoxicity of Cratoxylum Formosum Subsp. Pruniflorum Gogel Extracts in Oral Cancer Cell Lines

  • Promraksa, Bundit;Daduang, Jureerut;Chaiyarit, Ponlatham;Tavichakorntrakool, Ratree;Khampitak, Tueanjit;Rattanata, Narintorn;Tangrassameeprasert, Roongpet;Boonsiri, Patcharee
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7155-7159
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    • 2015
  • Background: Oral cancer is a health problem in Thailand. Cratoxylum formosum subsp. pruniflorum Gogel (Teawdang), normally consumed in northeast Thailand, has proven cytotoxic to cervical cancer cell lines including HeLa, SiHa and C-33A. Recently, Asian oral cancer cell lines, ORL-48 and ORL-136, were established. Therefore, we aimed to study cytotoxicity of Teawdang in these. Total phenolic (TPC) and flavonoid content (TFC), and antioxidant activity of Teawdang were also determined. Materials and Methods: Teawdang was purchased from Khon Kaen market during June-October 2013. Hexane (CHE), ethyl acetate (CEE) and methanol (CME) extracts of its edible part were analyzed for TPC by the folin-ciocalteau method and for TFC by an aluminium colorimetric method. Antioxidant activity and cytotoxicity in normal Vero cells and oral cancer cells were investigated. Cell viability was assessed using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Results: CME and CEE had higher TPC and TFC and antioxidant activity than CHE. Both CME and CEE, at $200{\mu}g$ dry wt/mL, were cytotoxic to the studied oral cancer cell lines. However, CME was cytotoxic to Vero cells whereas CEE was not. Compared to Vero cells, CEE significantly inhibited ORL-48 and ORL-136 growth (p=0.03 and p=0.02, respectively). Conclusions: CEE exhibited cytotoxic effects on the studied oral cancer cell lines but not normal Vero cells. The bioactive compounds in CEE should be further purified and elucidated for their mechanisms of action for development as anticancer agents.

Improvement of Porcine Epidemic Diarrhea Disease Vaccine Productivity by Ammonium Ion Removal in a Carberry Type Bioreactor (Carberry Type 생물반응기에서 암모늄 이온 제거에 의한 돼지유행성설사병 바이러스 백신 생산성 증대)

  • Lee, Chang-Jin;Jeong, Yeon-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.5
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    • pp.588-593
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    • 2011
  • The porcine epidemic diarrhea virus(PEDV) production yield in spinner flask cultures using Vero cells immobilized on microcarriers was improved by the selective adsorption of ammonium ions in a Carberry type bioreactor which was equipped with Phillipsite-Gismondine synthetic zeolite. Though the apparent cell growth seemed to be lower than that of control due to the aggregation of microcarriers between impeller shaft and the adsorbent, zeolite was found to not to be toxic to Vero cell, considering estimated glucose and lactate changes. Zeolite was observed to remove ammonium ions effectively in both steps of cell growth and virus production. In virus production, the virus titer with zeolite was two times higher than that without zeolite. Consequently, zeolite was found to be an ideal adsorbent for higher production of virus vaccine with the effective removal of ammonium ions.