• 제목/요약/키워드: Virus culture

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Efficient Diagnosis of Cucumber Green Mottle Mosaic Virus in Watermelon Using RT-PCR and Cloning of Coat Protein Gene (RT-PCR을 이용한 수박 Cucumber Green Mottle Mosaic Virus의 효율적인 진단 및 외피단백질 유전자의 클로닝)

  • 양덕춘;이진숙;김두욱;임용표;민병훈
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.519-524
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    • 1998
  • A simple and reliable method to diagnose cucumber green mottle mosaic virus of watermelon in Korea (CGMMV-WK) was determined by RT-PCR, and coat protein gene for CGMMV-WK was cloned. Comparing to a method reported by Lee et al. (1996), the method developed here showed a better RT-PCR reaction. RT-PCR was possible by one step in the PCR reaction mixture that contains 20 pmol of primer, reverse transcriptase (30 unit), RNasin (5 unit) using the crude RNA solution. RT-PCR condition for specifically diagnosing CGMMV-WK was that cDNA was synthesized at 42$^{\circ}C$ for 45 min followed by pre-denaturation at 95$^{\circ}C$ for 2 min, and then PCR reaction was carried out with a programmed condition that consisted of 36 sequential cycles at 96$^{\circ}C$ for 30 sec, 6$0^{\circ}C$ for 30 sec, and 72$^{\circ}C$ for 1 min. A gene encoding the coat protein of CGMMV-WK was cloned and characterized. Nucleotide sequence of coat protein gene of CGMMV-WK shared 98.77% and 99.38% of sequence identity with those of CGMMV-W and CGMMV-SH, respecitvely, however, all of amino acid sequences were same.

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A Study of Adenovirus Pneumonia: Early Summer Outbreak in 1996 (1996년 초여름에 유행한 Adenovirus폐렴의 검토)

  • Kim, Chang Keun;Chung, Churl Young;Kim, Jung Suk;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.145-153
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    • 1996
  • The incidence of Adenovirus(AV) pneumonia in children is estimated at 8%. Long-term complications reported with type 7 include hyperleucent lung syndrome, bronchiolitis obliterans and may require several months to recover. We reviewed 8 case of AV pnemonia confirmed by AV culture and sero-diagnosis among selected 19 AV pneumonia suspicious cases who were admitted to the Sanggye Paik Hospital of Inje University due to clinical pneumonia from May to July 1996. From 19 children, nasal aspirates or trachial aspirates were collected: viral agents were detected by virus isolation and virus antigen was detected by indirect immunofluorescent staining. Serologic diagnosis for type-specific AV antibody was done by neutralisation test. The results are as follows: 1) Age of the patients ranged from 6 months to 47 months with a mean of $24{\pm}9$ months. 6 were male and 2 were female(M:F= 3:1). 2) Monthly distribution showed 4 cases on June, 2 cases on May and July, respectively. 3) The mean duration of admission was 20.4 days and mean duration of fever was 11.3 days 4) Peripheral leukocyte counts were elevated in 4 cases(50%) and findings of shift to left were showed in 4 cases(50%) and serum aminotransferase activities were elevated in 6 case(75%). 5) Radiologic findings include linear and streaky infiltration(1/8 cases), patchy and lobar consolidation (7/8 cases), pleural effusion(2/8 cases) and hyperleucency was seen in 3/8 cases at time of admission and increased to 6/8 cases on hospital discharge. 6) The AV serotypes were type 7, type 3, and type 1. 7) Oxygen supply was done in 4 cases(50%) and all 8 patients include 2 cases of type 7 treated with artificial ventilator were survived. In conclusion, the serotypes of AV pneumonia outbreaks in early summer of 1996 were type 7, 3, 1 and, as showed severe clinical course, continued follow up surveillance is necessary to the long term pulmonary complications.

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Endometrial Cell Culture: Isolation, Characterization, and Immortalization (자궁내막 세포의 분리 및 배양 그리고 불멸화된 세포주의 확립)

  • Hong, In-Sun;Kim, Seok-Hyun;Koong, Mi-Kyoung;Jun, Jin-Hyun;Lee, Yong-Soon;Kang, Kyung-Sun
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.4
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    • pp.317-324
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    • 2003
  • 목 적: 본 실험의 목적은 자궁내막세포를 분리 및 배양법 확립과 함께 불멸화 시키는 것이다. 방 법: 자궁내막에서 상피세포(epithelial cells)와 기질세포(stromal cells)의 분리는 Satyawaroop 등(1979)의 방법에 기초를 두었다. 자궁내막에서 상피세포와 기질세포의 순수 분리도를 확인하고, 불멸화된 기질세포에서 SV40 large T antigen을 확인하기 위하여 면역형광 염색(immunocytochemistry)과 Western blot 기법을 이용하였다. 정상 기질세포의 경우 subconfluence (60%) 상태에서 transfection을 진행하였다. 순수 분리된 plasmid DNA와 Qiagen 사의 superfect를 이용하여 transfection을 실시하였다. 결 과: 본 연구에서 우리는 두 가지 형태의 자궁내막 세포의 분리 및 배양에 성공하였다. 상피세포는 다면체의 형태를 띠며, 선(grandular)조직의 조각으로부터 나선형으로 자란다.기질 세포는 길쭉한 형태를 띠며, 상피세포에 비해 오래 살고, 빠르게 증식하여 나란한 형태로 배열된 세포 다발(cell bundle)을 형성한다. 이렇게 분리된 세포들은 95%의 균질성을 보였으며, 면역형광염색과 western blot을 통해 확인 하였다. 한편 SV40(Simian Virus 40) large T 항원을 암호화 하고 있는 염기 서열을 포함한 플라스미드 벡터로 안정적인 트랜스펙션을 시킴으로써 불멸화 된 자궁내막의 기질 세포주를 확립하였다. 불멸화 된 세포는 그 세포가 유래한 정상의 세포와 동일한 표현형을 가지고 있었다. 결 론: 본 연구에서, 우리는 자궁내막에서 상피세포(epithelial cells)과 기질세포(stromal cells)를 분리하여 배양법을 확립하였다. 동시에 SV40 large T antigen을 이용하여 불멸화된 세포주를 확립하였다. 이렇게 확립된 세포주는 자궁의 생리작용 연구 및 자궁내막증(Endometriosis)과 자궁암(Endometrial cancer) 등과 같은 여러 자궁관련 질병 연구에 많은 도움이 될 것으로 사료된다.

Gene Expression in The Fifth Generation of TMV Resistant Transgenic Tobacco Plane at Elevated Temperature (TMV 저항성 형질전환 연초식물체 제 5 세대에서 유전자 안정성 및 고온조건에서의 유전자 발현)

  • 이기원;박성원;이청호;박은경;김상석;최순용
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.245-250
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    • 1998
  • Tobacco mosaic virus(TMV) coat protein cDNA was transformed to Nicotiana tabacum cv. NC82 and the transgenic tobacco plants resistant to TMV infection were isolated in the next generation. The expression of TMV coat protein cDNA and genetic stability of the fifth generation of TMV resistant transgenic tobacco plants at the higher temperature were investigated. The TMV coat protein cDNA was amplified by genomic PCR in all the TMV resistant transgenic tobacco plants. The TMV coat protein expressed in the transgenic tobacco plants was detected at very low level by immunoblot hybridization. Even in tansgenic plants that showed the viral symptom only on very late sucker growth (delay type plants), the coat protein expression in the suckers was much less than that of susceptible tobacco infected with TMV. The TMV coat protein expressed in the transgenic tobacco plants was below 0.01% of total protein. Transcription and expression of the coat protein cDNA in delay type plants were observbed at high temperature (38$^{\circ}C$), and TMV replication was suppressed at both 28$^{\circ}C$ and 38$^{\circ}C$. This indicates that unlike the resistance conferred by 'N' gene. TMV resistance of transgenic tobacco plant won't break down at high temperature.

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Microbiological Safety and Tensile Strength of Gamma Irradiated Porcine Tendon for the Development of Xenografts (이종이식재 개발을 위한 감마선 조사 돼지 인대의 미생물학적 안전성 및 인장강도)

  • Kim, Jeongsoo;Sung, Nak-Yun;Jo, Eu-Ri;Choi, Jong-il;Song, Beom-Seok;Kim, Jaekyung;Lim, Youn-Mook;Lee, Ju-Woon;Lee, Kwang-Won;Kim, Tae-Woon;Byun, Myung-Woo;Kim, Jae-Hun
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.75-80
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    • 2011
  • This study was to determine the microbiological safety and tensile strength of gammairradiated porcine tendon for the development of safe xenografts. Escherichia coli and Bacillus subtilis were used as model pathogens and inoculated as $10^6{\sim}10^7log$ colonies forming unit $(CFU)g^{-1}$. As model virus from porcine, porcine parvovirus (PPV), bovine viral diarrhoea virus (BVDV) and poliovirus were inoculated as $10^5{\sim}10^6$ tissue culture infectious dose $(TCID)_{50}g^{-1}$ into porcine skin. The $D_{10}$ value of E. coli and B. subtilis was measured as $0.32{\pm}0.082kGy$ and $4.0{\pm}0.312kGy$, respectively. Additionally, the $D_{10}$ values of PPV, BVDV and poliovirus were also shown as $1.75{\pm}0.131kGy$, $3.70{\pm}0.212kGy$ and $6.26{\pm}0.332kGy$, respectively. Gamma irradiation decreased the tensile strength of porcine tendon. Results indicate that microbiological safety of porcine tendon can be improved significantly by gamma irradiation. However, further studies are needed to improve the tensile strength of gamma-irradiated porcine tendon.

Filtration Efficiencies of Commercial Face Masks in Korea for Biological Aerosols (국내 출시 마스크의 바이오에어로졸 여과효율 평가)

  • Choi, Sueun;Choi, Doseon;Jang, Sung Jae;Park, SungJun;Yoon, Chungsik;Lee, Kiyoung;Ko, GwangPyo;Lee, Cheonghoon
    • Journal of Environmental Health Sciences
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    • v.48 no.2
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    • pp.116-122
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    • 2022
  • Background: The recent COVID-19 pandemic is one of the worst disease outbreaks of the 21th century. Due to a lack of reliable antiviral therapeutics, wearing face masks is recommended to prevent airborne infection originating from virus-contaminated bioaerosols. Objectives: The aim of this study was to evaluate the filtration efficiencies of face masks that are commercially available in South Korea for a biological aerosol of Staphylococcus aureus (S. aureus) and murine coronavirus, a well-known surrogate for human coronaviruses. Methods: We collected six different kinds of commercial masks: two Korea Filter (KF)94 (KF94-1, KF94-2) masks, one surgical (Surgical-1) mask, one anti-droplet (KF-AD-1) mask, and two dust (Dust-1, Dust-2) face masks. S. aureus (ATCC 6538), a well-performing test bacteria and murine coronavirus (ATCC VR-764) were prepared under a suitable culture condition. Then, a mask biological filtration tester was used to examine the microbial filtration efficiencies of masks. Test microorganisms were quantitatively measured via cultivation methods and microbial filtration efficiencies were calculated appropriately. Results: All face masks showed over 99.6% filtration efficiency for S. aureus or murine coronavirus. There were no significant differences among the bacterial filtration efficiencies of the face masks. KF94-1 (99.97±0.08%) and Dust-1 mask (99.97±0.07%) showed the highest (over 99.9%) filtration efficiency for murine coronavirus. KF94-1 or Dust-1 masks showed a significant virus filtration efficiency compared to Surgical-1 mask (p<0.05; Mann-Whitney U test). Conclusions: All the commercially available face masks used in this study can filter S. aureus or murine coronavirus in bioaerosols efficiently, regardless of the mask type. Therefore, our results suggest that wearing a certified face mask is a reliable means to prevent the transmission of infectious airborne diseases via biological aerosols.

Characteristics of HIV-Tat Protein Transduction Domain

  • Yoon Jong-Sub;Jung Yong-Tae;Hong Seong-Karp;Kim Sun-Hwa;Shin Min-Chul;Lee Dong-Gun;Shin Wan-Shik;Min Woo-Sung;Paik Soon-Young
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.328-335
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    • 2004
  • The human immunodeficiency virus type 1 (HIV-I) Tat protein transduction domain (PTD), which con­tains rich arginine and lysine residues, is responsible for the highly efficient transduction of protein through the plasma membrane. In addition, it can be secreted from infected cells and has the ability to enter neighboring cells. When the PTD of Tat is fused to proteins and exogenously added to cells, the fusion protein can cross plasma membranes. Recent reports indicate that the endogenously expressed Tat fusion protein can demonstrate biodistribution of several proteins. However, intercellular transport and protein transduction have not been observed in some studies. Therefore, this study exam­ined the intercellular transport and protein transduction of the Tat protein. The results showed no evi­dence of intercellular transport (biodistribution) in a cell culture. Instead, the Tat fusion peptides were found to have a significant effect on the transduction and intercellular localization properties. This sug­gests that the HIV-1 PTD passes through the plasma membrane in one direction.

Subjectivity toward Death among HIV-Positive Men (HIV 감염인 남성의 죽음에 대한 주관성)

  • Lee, Eun-Ju
    • Journal of Hospice and Palliative Care
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    • v.17 no.3
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    • pp.142-150
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    • 2014
  • Purpose: This study was to identify the attitude of Korean HIV (Human Immunodeficiency Virus)-positive men toward death. Methods: A Q-methodology was performed with 20 HIV-positive male individuals. Participants were asked to select and answer questions among a set of 40 Q-statements using a 9-point scale. The collected data were analyzed using the PC QUANL program. Results: Participants' attitudes toward death were categorized into four types. Type I was characterized by respect for life, type II by reality orientation, type III by pain evasion and type IV religious beliefs. Conclusion: It is necessary to develop an assessment tool and an intervention program for HIV-positive individuals.

Immortalization of Rabbit Corneal Fibroblasts by Overexpression of Simian Virus 40 Large T antigen

  • Cho, Seung-Ju;Park, Yuk-Pheel;Lim, Heon-Man;Kim, Jae-Chan;Yang, Eun-Kyung;Park, Jung-Keug;Yoon, Do-Young;Lee, Hee-Gu
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.85-92
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    • 2004
  • Immortalization of primary corneal cells has influence on pharmacy, medical and biological fields. Especially, investigation of immortalization mechanism using viral oncoproteins is useful for medical treatments, and these cell lines will be useful materials for toxic test of medical supplies and cell biological experiments. Rabbit corneal fibroblasts in culture undergo a finite number of divisions before they reach a terminally non-proliferating state known as replicative senescence. Therefore, we attempted to induce immortalization of rabbit corneal fibroblasts with SV 40 large T antigen. As a result of experiment, expression of SV 40 large T antigen was confirmed, and expression of proteins related to cell cycle repressor was decreased in the transfection group compared with non-transfection group. According to the results of cell cycle phase distribution test, SV 40 large T antigen-transfected cells had obtained higher proliferation rate than primary cells. It was confirmed that during induction of immortalization, SV 40 large T antigen was not able to increase telomerase activity. In conclusion, we made a rabbit corneal fibroblast cell line with SV40 large T antigen. This cell line will be useful for further studies of mammalian fibroblast biology, particularly with regard to angiogenesis and malignant transformation. In addition, this cell line offers opportunity for testing potential therapeutics and can be used for toxicity tests of materials or cosmetics. In the future, our cell line can potentially be utilized in a wide range of biology related fields.

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Cytomegalovirus Infection in Pediatric Renal Transplant Recipients: A Single Center Experience

  • Kim, Mi Jin;You, Ji Hye;Yeh, Hye Ryun;Lee, Jin A;Lee, Joo Hoon;Park, Young Seo
    • Childhood Kidney Diseases
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    • v.21 no.2
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    • pp.75-80
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    • 2017
  • Purpose: To investigate the frequency, presentation, management, and outcome of cytomegalovirus (CMV) infection in pediatric patients who underwent renal transplantation. Methods: We performed a retrospective chart review of 70 patients under the age of 18, who underwent renal transplantation between January 1990 and November 2014. A diagnosis of CMV infection was based on serology, molecular assays, antigenemia assays, and culture. CMV infection was defined as detection of virus and CMV disease was diagnosed when clinical signs and symptoms were present. Results: The number of patients with CMV infection was 18 (25.7% of renal transplant recipients). Twelve were male (66.7%), and the $mean{\pm}standard$ deviation (SD) age at infection was $13.3{\pm}3.9$ years. Median time of infection after renal transplantation was 4 months (range 1.0-31.0 months). Pretransplantation CMV status in the infected group was as follows: donor (D)+/recipient (R)+, 11 (61.1%); D+/R-, 7 (38.9%); D-/R+, 0; and D-/R- 0. Nine patients had CMV disease with fever, leukopenia, thrombocytopenia, or organ involvement such as enteritis, hepatitis, and pneumonitis. The age of disease occurrence was $13.1{\pm}3.9$ years and the median time to disease onset after renal transplantation was 8 months (range 1.0-31.0). Immunosuppressive agents were reduced or discontinued in 14 patients (77.8%), antiviral agents were used in 11 patients (61.1%), and all patients with CMV infection were controlled. Conclusions: A quarter of the patients had CMV infection about 4 months after renal transplantation. CMV infection was successfully treated with reduction of immunosuppressants or with antiviral agents.