• Title/Summary/Keyword: Virus culture

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Regeneration of Fertile Transgenic Rice Plane from a Korean Cultivar, Nakdongbyeo (한국 재배종 낙동벼에서 임성 형질전환식물체의 재분화)

  • Soo In LEE;Hyun Jin CHUN;Chae Oh LIM;Jeong Dong BAHK;Moo Je CHO
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.175-182
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    • 1995
  • Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.

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Study on the Winter Mass Mortality of red sea bream, Pagrus major in South sea area (동절기 남해안 참돔(Pagrus major)의 대량폐사에 관한 연구)

  • Choi, Hye-Sung;Jung, Sung-Hee;Hur, Young-baek;Yang, Jun-Yong
    • Journal of fish pathology
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    • v.21 no.1
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    • pp.35-43
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    • 2008
  • Pathological symptoms and hematological parameters of red sea bream, Pagrus major and water temperature in the culture ground was investigated to clarlify the cause of winter moratility. Dead fish showed green liver and accumulation of ascites in the cavity. A few Bivagina tai were also found on the gill but either bacteria or virus were not. When hematological parameters from fish taken before/after the winter mortality were compared, blood glucose, serum AST and ALT, and total cholesterol, trigyceride and total protein were significantly decreased in the fish after the winter mortality. These results may explain that the nutritional level of fish was decreased because fish could not fed during winter season. According to the CORI monitoring system operating by KODC, NFRDI a long term water temperature from Dec. 25, 2005 to Feb. 24, 2006 (60 days) were exposed the low water temperature environments to the red sea bream. First of all, mass mortality began at Sarayngdo where low temperature below 8℃ continued for 42 days. The winter mortality did not occured in the depths of 9 m to 19.2 m where difference of water temperature in the surface and bottom was only within 0.1℃. But in the depth of 7.5 m to 11 m winter mortality occured where water temperature in the surface and bottom showed much variation ranged from 0.2℃ to 1.4℃. From these results, great difference of water temperature in the surface and bottom of the culturing area might results in winter mortality of red sea bream.

The antibacterial effect of photo-catalytic titanium dioxide on canine skin (광 촉매 이산화 티타늄의 개 피부에 대한 항균효과)

  • Chang, Hwa-Seok;Kim, Ji-Eun;Chung, Dai-Jung;Lee, Jung-Sun;Choi, Chi-Bong;Kim, Hwi-Yool
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.279-284
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    • 2006
  • Photo-catalytic products have been widely used at home and hospital to prevent bacteria, virus and fungus. Activities of anti-bacteria, anti-viruses and anti-fungi are based upon direct contact of crystals and particles of titanium dioxide with pathogens, into which titanium is catalyzed by photo. Those antimicrobial activities of the photo-catalytic titanium dioxide have been proved in vitro. However, in vivo tests of those activities have not been carried out on dog skin. Aim of this study was to evaluate the antimicrobial activities of the catalytic titanium dioxide in vivo. Ten beagle dogs were divided into two groups. One group was sprayed with 10ml of titanium dioxide(1 mg/ml) whereas the other was not. The treated dogs were exposed under the sunlight for 120 min. A set of three hairs was taken 15, 30, 60 and 120 min after the exposure and the bacteria contaminated in hairs were amplified in, Muller Hilton broth at $35^{\circ}C{\pm}1$ for 3 h. The supernatant of the bacterial culture was diluted 1 : 10 in phosphafe-buffered saline. One milliliter of the diluents was transferred into triphenyltetrazolium medium(TTC) and incubated at $35^{\circ}C{\pm}1$ for 2 days. The number of bacteria was counted. The number of bacteria colonies was decreased compared to control group. To further investigate the longevity effect of titanium dioxide, the dogs were kept in indoor without sun light for 6 and 12 h, 1, 2, 3, 7, 14 days after exposure of the chemical during each 15, 30, 60 min. The number of bacteria colony in 1ml was counted. The number of bacterial colonies was decreased. Treated group is exposured by sun light during 15 min, the longevity effect of titanium dioxide is continued by 1 week. Treated group is exposured by sun light during 30, 60 min, the longevity effect of titanium dioxide is continued over 2 weeks. These data indicated that the photo-catalytic titanium dioxide may be used for prevent bacteria on dog skin.

One step multiplex RT-PCR preventing DNA carryover contamination for differential diagnosis of swine influenza viruses (DNA 교차 오염 방지 기능을 가진 돼지 인플루엔자바이러스 감별진단용 one-step multiplex RT-PCR 진단법)

  • Kim, Hee-Jung;Kim, Eun-Mi;Shin, Yeun-Kyung;Song, Jae-Young;Kim, Seong-Hee;Lee, Kyoung-Ki;Lee, Myoung-Heon;Kim, Young-Hwa;Park, Jun-Cheol;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.263-271
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    • 2014
  • In this study, we developed a cost and time saving one-step multiplex RT-PCR for the simultaneous detection and differentiation of swine influenza viruses (SIV) and 2009 pandemic influenza H1N1 virus (pH1N1). The one-step multiplex RT-PCR using four sets of primer was confirmed to be capable of detection of all SIV subtypes and differential diagnosis of major SIV subtype H1, H3 and pH1N1 on individual or mixed viral culture samples. The sensitivity of the multiplex RT-PCR was determined to be at least $2^{-6}$ $HA/25{\mu}L$ of the presented SIVs, providing sufficient efficacy for a routine SIV monitoring in diagnostic laboratories. In addition, compared with the conventional RT-PCR methods that cannot avoid the carryover DNA contamination, the developed RT-PCR applied with the uracil DNA glycosylase (UNG) system was proven to prevent a false positive reaction by carryover contamination of the pre-amplified DNA. In conclusion, the one-step RT-PCR with UNG system could be applicable to detect and differentiate of SIV from the viral cultures without worry of carryover DNA contamination in clinical laboratories.

Determination of the Optimal Concentration of Fetal Bovine Serum for the Growth of Two Insect Cell and Viruses (두 가지 곤충 세포주에 대한 배양 및 바이러스 증식을 위한 최적 FBS 농도 결정)

  • Lee, Jae-Kyung;Koo, Hyun-Na;Woo, Soo-Dong
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.319-324
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    • 2007
  • To determine the optimal concentration of fetal bovine serum (FBS) on the growth of insect cells and the multiplicity of viruses, the growth of cells (Sf21 and Bm5) and viruses were examined on the various concentrations of FBS. In view of the viability, growth speed, proliferation of cells and the amount of FBS, the most proper concentration for the cell culture were 7% and 5% for Sf21 and Bm5, respectively. The multiplicity of viruses at the various concentrations of FBS was similar in both cell lines at 5 days post-infection (p.i.). However, it differed significantly at 2 and 3 days p.i. The proper concentration of FBS were 10% and 3% for Sf21 at 2 and 3 days p.i., respectively, and 5% for Bm5 at both 2 and 3 days p.i. These results suggested that the optimal concentration of FBS should be determined according to the used cell lines and viruses for their optimum production.

A study on the effect of entrepreneurship and CEO competency on management performance of SME managers in the service economy era (서비스경제시대 중소기업 경영자의 기업가지향성과 CEO역량이 경영성과에 미치는 영향 연구)

  • An, Sehong;Kim, Hyunsoo
    • Journal of Service Research and Studies
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    • v.10 no.3
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    • pp.81-101
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    • 2020
  • Humanity has reached the age of service economy. A new economy requires new management. The chief executive of new management is the CEO. The results of management decision-making are reflected in the company's performance, and are the basis for future growth engines. This study empirically analyzed the effect of CEO competence, corporate orientation, and management performance of small and medium-sized mid-sized business managers on each other. In the study, a hypothesis was established between eight variables. In addition, the relationship between corporate orientation and CEO competency, which has not been studied, was also identified. Set personal effectiveness, willingness to innovate, opportunity recognition, information perception, relationship formation, business management, leadership, organizational culture, and human resource management as measures of CEO competency. After defining the operation in accordance with this study, the analysis was conducted. As a result of the analysis, corporate orientation did not produce a meaningful result in management performance. And CEO competency showed a positive (+) effect on management performance. It can be judged that the CEO variable, a management variable, had a positive effect on management performance. Also, there was a significant result in the relationship between corporate orientation and CEO competency. Innovativeness, initiative, and risk-taking have positive implications for CEO competency. Lastly, corporate orientation showed statistically significant results on management performance through the mediation of CEO competency. It is worth noting the results of this study in that the time of the study was the timing of the global pandemic (fendermic) of the COVID19 virus. In the future, in-depth research is needed on the relationship between CEO-related factors and management performance in a more changed economic environment.

Regulation of hPTH Expression In Virto Using the Tetracycline Inducible Retrovirus Vector System (Tetracycline Inducible Retrovirus Vector System을 이용한 In Vitro에서의 인간 부갑상선 호르몬의 발현 조절)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.157-162
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    • 2006
  • Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

A Study of the Retrovirus-Mediated Transgenic Chicken Production on Chicken Embryos (닭 수정란에서 Retrovirus를 이용한 형질전환 닭 생산 연구)

  • Byun S. J.;Park C.;Kim S. W.;Park J. K.;Chang W. K.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.225-229
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    • 2005
  • Microinjection of recombinant retrovirus beneath the blastoderm of non-incubated chicken embryo is now the most widespread method for generating transgenic chickens, but transgenesis rates are very low. So to improve this problem, we first introduced retrovirus vector carrying RSV-GFP gene to an one-cell embryo culture system. To investigate whether retrovirus could work on an one-cell chicken embryo, we microinjected the concentrated retrovirus stocks into the germinal disc of one cell or stage-X chicken embryos. Analysis of reporter gene expression on day 4 embryos showed that GFP expression was observed in the only stage-X chicken embryo but was not in the one-cell embryo group. These results suggest that retrovirus system is the most efficient method to generate transgenic chickens in the stage-X embryo.

Cigarette Smoke Extract-Treated Mouse Airway Epithelial Cells-Derived Exosomal LncRNA MEG3 Promotes M1 Macrophage Polarization and Pyroptosis in Chronic Obstructive Pulmonary Disease by Upregulating TREM-1 via m6A Methylation

  • Lijing Wang;Qiao Yu;Jian Xiao;Qiong Chen;Min Fang;Hongjun Zhao
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.3.1-3.23
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    • 2024
  • Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages. In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68+ cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO-CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.

Microbiological Identification and Distribution of Metal Components in Suspended Particulate Matter during Yellow Sand Phenomena at TaeAn Region in 2003 (2003년 태안지역에서 황사 부유분진의 미생물학적 동정과 금속 성분 및 농도)

  • Bae, Kang Woo;Kim, Jong Ho;Kim, Youn Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.2
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    • pp.167-173
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    • 2005
  • Background : Airborne particles during Yellow Sand phenomena are known to be associated with the respiratory disease. The purpose of this study was to evaluate the concentration and metal component properties of Yellow Sand particles and compare with airborne microbial concentration and species in non Yellow Sand and Yellow Sand phenomena. Methods : Samplings were carried out in 2002 in Seosan, during non Yellow Sand and Yellow Sand phenomena. Samples were taken using the 8-stage Cascade impactor and metallic elements were analyzed by XRF. Those were culture on the media for bacterial and fungal culture and celline for virus. Results : The concentration of total suspended particulate matter were respectively $80.2{\mu}g/m^3$, $40.3{\mu}g/m^3$ in non Yellow Sand and Yellow Sand phenomena. The concentration of metallic elements such as Ca, Fe, Cu and Zn in Yellow Sand phenomena were higher than its in non Yellow Sand. Two bacteria, Bacillus species and Staphylococcus were grown in two periods. In both periods, several fungal spores(Mucor species, Cladosporum, Alternaria, Aspergillus, Penicillium, and Alternaria species) were identified. The differences of bacteria and fungus species not observed in Yellow Sand and non Yellow Sand. Any viruses were not isolated in between both periods. Conclusions : The concentration of total suspended particulate matter and some metallic elements in Yellow Sand phenomena were higher than its in non Yellow Sand. The difference of bacteria and fungus species was not observed in non Yellow Sand and Yellow Sand phenomena.