• Title/Summary/Keyword: virus-free

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A Gene Encoding Phosphatidyl Inositol-specific Phospholipase C form Cryphonectria parasitica Modulates the Hypoviral-modulated Laccase1 Expression

  • Kim, Dae-Hyuk
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.159-161
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    • 2005
  • Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.

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Pathogenicity of a Local Isolate of Chicken Anemia Agent for Chickens and Prevalence of Antibody in Chicken Flocks (국내분리 chicken anemia agent의 닭에 대한 병원성과 야외계군의 항체 보유상황)

  • 김선중
    • Korean Journal of Poultry Science
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    • v.18 no.3
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    • pp.141-150
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    • 1991
  • A local isolate of chicken anemia agent (CAA), isolate 89-69. was tested for pathogenicity for chickens. When chickens from a specific pathogen free (SPF) flock were inoculated intramuscularly with the isolate at one day old, all the chickens showed severe anemia at 14 to 18 days post inoculation(DPI) and returned to normal at 25DPI, Some of the inoculated chickens (27∼33%) died between 13 to 17 DPI's with lesions of severe aplasia of bone marrow and thymic atrophy. In chickens kept in contact with inoculated chickens, some of the chickens had anemia at 25 and 28 DPI's. Virus could be reisolated from inoculated as well as in contact chickens till 21 DPI. Antibodies to CAA could be detected in all inoculated and in contact chickens when tested at 42 DPI by the indirect fluorescent antibody method. When chickens from a different SPF flock were inoculated at one day old, degrees of anemia, both in frequency of incidence and severity, were low These chickens were proved partly to have antibodies to CAA when tested for hatchmates. In a survey for antibodies to CAA in field chicken flocks, one out of 7 flocks(14%) aged 3 to 10weeks was antibody positive whereas 19 out of 20 flocks(95%) over 20 weeks of age were positive. Altogether 29 out of 39 flocks (74%) were antibody positive.

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Microtuberization and Acclimatization in the Dioscorea cayenensis Thunb. by the Carbon Source (탄소급원에 의한 얌의 기내 비대근 형성과 순화)

  • Lee, Na Nyum;Kim, Ji Ah;Kim, Yong Wook;Kim, Tae Dong
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.125-130
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    • 2018
  • In this experiment, we investigated the effects of various carbon sources and concentrations on the microtuber induction and acclimatization of the yam (Dioscorea cayenensis). First, the effects of the in vitro carbon sources and concentrations on the microtuber induction were examined. The highest efficiency of the microtuber induction was obtained in the 7% sucrose treatment, whereas the glucose treatment shows no effect on the microtuber formation. Secondly, the effects of the survival rate and the microtuber formation rate after the acclimatization were examined. The diameter (6.1 mm) and fresh weight (0.5g) of the tuberous root are the highest in the pretreatment of the 7% sucrose. Although the survival rate of the pretreatment of the low concentration sucrose (3% sucrose) is 100 %, the growth and development were inhibited. These results suggest the 7% sucrose treatment is appropriate for the yam microtuber formation and acclimatization. In addition, this protocol could be used for the propagation of virus- or disease-free clones and the multiplication of elite yam cultivars.

Effects of Foliar Application of Chitosan and Wood Extraction on Rooting and Tuber Formation of Plug Seedlings in Potatoes (Chitosan과 목초액 엽면살포에 의한 감자 플러그 삽목표의 발근 및 괴경형성효율)

  • 송창길;강태균
    • Korean Journal of Organic Agriculture
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    • v.8 no.1
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    • pp.89-99
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    • 1999
  • To do mass multiplication of plug seedlings in potatoes, apical stem cuttings originated from virus-free microtubers were cut to one-two internodes and transplanted into the plug-tray. After 10days, we applied Chitosan and Wood Extraction on rooting and tuber formation of plug seedlings. To improve field adaptability of plug seedlings, rooted cuttings with a height of 20cm after 20days of cutting were transplanted ito the fields, We applied 500~2000ppm Chitosan on growth characteristics and tuber formation of that. The above and underground growths, such as plant height and number of leaves were significantly more vigorous after treatment with 500~1,000ppm Chitosan and 2,000ppm Wood Extraction, the spray treatment was carried out five times at intervals of four days after ten days of transplanting. T-N, K, P, Mg and Na, were higher as the concentrations of chitosan and Wood Extraction were higher. The growth and tuber yield in plug seedlings planting plot and seed potatoes planting plot were effectively highter as foliar application of Chitosan(500~2,000ppm) was done after planting the plot. T-N content in leaves and tuber was higher as the concentration of Chitosan was high. A similar tendency was shown in K, P and Mg. In the small tuber(under 30g), the number of tubers and tuber yield were relatively increased in the seed potatoes planting plot, but the large tubers(over 80g) yield was higher in the plug seedlings planting plot, and in order to increase tuber yield in plug seedlings it was necessary to add plant density to the field.

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Combined Effects of Six Cytokine Gene Polymorphisms and SNP-SNP Interactions on Hepatocellular Carcinoma Risk in Southern Guangxi, China

  • Bei, Chun-Hua;Bai, Hua;Yu, Hong-Ping;Yang, Yan;Liang, Qing-Qing;Deng, Ying-Ying;Tan, Sheng-Kui;Qiu, Xiao-Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6961-6967
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    • 2014
  • Cytokine gene single nucleotide polymorphisms (SNPs) are involved in the genesis and progression of hepatocellular carcinoma (HCC). We hypothesized that combined effects of cytokine gene SNPs and SNP-SNP interactions are associated with HCC risk. Six SNPs in cytokine genes (IL-2, IFN-${\gamma}$, IL-$1{\beta}$, IL-6, and IL-10) were genotyped in a study of 720 Chinese HCC cases and 784 cancer-free controls. Although none of these SNPs individually had a significant effect on the risk of HCC, we found that the combined effects of these six SNPs may contribute to HCC risk (OR=1.821, 95% CI=1.078-3.075). This risk was pronounced among smokers, drinkers, and hepatitis B virus carriers. A SNP-SNP interaction between IL-2-330 and IFN-${\gamma}$-1615 was associated with an increased HCC risk (OR=1.078, 95% CI=1.022-1.136). In conclusion, combined effects of SNPs and SNP-SNP interactions in cytokine genes may contribute to HCC risk.

Role of radiotherapy in local control of non-AIDS associated Kaposi's sarcoma patients in Korea: a single institution experience

  • Chang, Ji Hyun;Kim, Il Han
    • Radiation Oncology Journal
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    • v.30 no.4
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    • pp.153-157
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    • 2012
  • Purpose: There has been no definite consensus on standard treatment, either local or systemic, for the Kaposi's sarcoma (KS). Radiotherapy (RT) can be a good local therapeutic choice especially in non-AIDS associated KS (NAKS) for its indolent behavior. Materials and Methods: Medical records of 17 KS patients treated with RT at the Seoul National University Hospital from February 1998 to January 2012 were retrospectively reviewed. One human immunodeficiency virus (HIV)+ patient with 3 lesions was excluded. The total number of the lesion was 23 among the 16 patients. The median follow-up period was 27.9 months. Correlation between response and variables was analyzed using the logistic regression model. Median age of the patients was 75 years. All the 23 lesions were located at the extremities. Fourteen (61%) of those had pain or local swelling as the initial presentation. Ten patients had possible causes of immunodeficiency and were regarded as iatrogenic, and other 6 were classic KS. Median dose of RT was 36 Gy. Results: No KS-related death was observed. Excluding 2 with short-term follow-up only, complete response and partial response were obtained in 2 (9%) and 19 (73%) lesions, respectively. Of those, 3 lesions underwent local progression. Six had out-of-field recurrence after RT. Symptom improvement was achieved in 13 (93%) of 14 patients. Grade 2 skin toxicities were found in 9 lesions but all got improvement after treatment. When divided into responsive and progressive group, free from progression was not related to any of the possible variables. Conclusion: RT is effective in local control of NAKS resulting great response rate.

Barriers to Health Service Utilization Among Iranian Female Sex Workers: A Qualitative Study

  • Asadi-AliAbadi, Mehran;Abolghasemi, Jamileh;Rimaz, Shahnaz;Majdzadeh, Reza;Rostami-Maskopaee, Fereshteh;Merghati-Khoei, Effat
    • Journal of Preventive Medicine and Public Health
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    • v.51 no.2
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    • pp.64-70
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    • 2018
  • Objectives: In most countries around the world, sex work is an illegal activity. Female sex workers (FSWs) in Iran hide their identities, and they are known to be a hard-to-reach population. Despite free access to HIV testing, fewer than half of FSWs receive HIV testing. The purpose of this study was to characterize the reasons for which FSWs do not seek testing at drop-in centers (DICs) and voluntary counseling and testing (VCT) centers in Iran. Methods: A qualitative study was conducted in 2016. The participants were 24 FSWs who received services at VCT centers and DICs for vulnerable females in the north of Iran and 9 males who were the clients of FSWs. In this study, we made use of purposive sampling and carried out a thematic analysis. Results: We found 4 major and 6 minor themes. The major themes were: fear of being infected (with HIV), stigma, indifference, and knowledge. Conclusions: Despite the significant efforts made by the government of Iran to establish and expand DICs for vulnerable females, the number of FSWs receiving services at these centers has not been very considerable. Consequently, by introducing and implementing training programs for peer groups, it may be possible to take steps toward establishing strategic programs for the control and prevention of HIV/AIDS.

Probiotic Property of Lactobacillus pentosus Miny-148 Isolated from Human Feces (인체분변으로부터 분리한 유산균 Lactobacillus pentosus Miny-148의 생균제 특성 연구)

  • Jung, Min-Young;Park, Yong-Ha;Kim, Hyun-Soo;Poo, Ha-Ryoung;Chang, Young-Hyo
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.177-184
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    • 2009
  • Three hundred lactic acid bacteria isolated from human feces were studied their probiotic characters to develop potential probiotics. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, thermal stability, antimicrobial, anticancer cell, and antiviral activity. Strain Miny-148 was selected as a potential probiotic bacterium which showed resistance to low pH, bile salts and thermal stability. On the basis of fatty acid profiles and 16S rDNA sequences analysis, the strain was identified as Lactobacillus pentosus (similarity 99.9%). The strain, L. pentosus Miny-148, showed broad antimicrobial spectrum against E. coli O157:H7, Shigella flexneri, Bacillus anthracis, Staphylococcus aureus, E. coli, Vibrio cholerae, V. vulnificus, Salmonella typhimurium, and Methicillin-resistant S. aureus (MRSA). Cell-free culture supernatant of the strain also inhibited against the growth of HT-29 colon cancer cell and transmissible gastroenterits virus.

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.

The Effect of Spent Medium Recycle on Cell Proliferation, Metabolism and Baculovirus Production by the Lepidopteran Se301 Cell Line Infected at Very Low MOI

  • Beas-Catena, Alba;Sanchez-Miron, Asterio;Garcia-Camacho, Francisco;Contreras-Gomez, Antonio;Molina-Grima, Emilio
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1747-1756
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    • 2013
  • The aim of this paper was to study the effect of spent medium recycle on Spodoptera exigua Se301 cell line proliferation, metabolism, and baculovirus production when grown in batch suspension cultures in Ex-Cell 420 serum-free medium. The results showed that the recycle of 20% of spent medium from a culture in mid-exponential growth phase improved growth relative to a control culture grown in fresh medium. Although both glucose and glutamine were still present at the end of the growth phase, glutamate was always completely exhausted. The pattern of the specific glucose and lactate consumption and production rates, as well as the specific glutamine and glutamate consumption rates, suggests a metabolic shift at spent medium recycle values of over 60%, with a decrease in the efficiency of glucose utilization and an increase in glutamate consumption to fuel energy metabolism. Baculovirus infection provoked a change in the metabolic pattern of Se301 cells, although a beneficial effect of spent medium recycle was also observed. Both growth rate and maximum viable cell density decreased relative to uninfected cultures. The efficiency of glucose utilization was dramatically reduced in those cultures containing the lowest percentages of spent medium, whereas glutamine and glutamate consumption was modulated, thereby suggesting that infected cells were devoted to virus replication, retaining their ability to incorporate the nutrients required to support viral replication. Recycle of 20% of spent medium increased baculovirus production by around 90%, thus showing the link between cell growth and baculovirus production.