• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.470-476
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• 1992

Upon lyophilization of Nocardia mediterranei, the effects of cryoprotectants, cell concentration and drying time on viability were examined, The data were treated by computer according to response surface analysis. As a result, the maximum value of presumed viability was 39.3% under the optimal conditions of 1l.6%(v/v) sucrose, $1.16{\times}10^{11}$(CFU/ml) cell concentration, and drying time for 6.18 hrs. We also used the starter cell of rehydrated solution after lyophilization in industrial production, obtained the fermentation pattern and the purity of rifamycin B which were the same with control (FVM) and it is possible for us to use N mediterranei as a starter cell after the storage of lyophilization for 18 months.

• Microbiology and Biotechnology Letters
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• v.8 no.2
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• pp.87-92
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• 1980

These experiments were conducted to manage more safe the selected strains and improve their characteristics. The results obtained were as follows: Preservation on soil at the range of 0 to 5 C was suitable and there were no remarkable changes on their abilities to produce citric acid until 10 months preservation. The successive transfer of spore slightly stimulated the selected strain, M-315 to produce citric acid and the spore precultured on sporulation medium for 7-10 days was desirable as inoculum. Under UV-irradiation from the selected strains, 109 mutants whose morphological characteristics were changed were isolated. Among them, the mutant M-80-12 was shown 3.2％ increase on acid producing ability than that of its parent.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.161-166
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• 1994

Since garlics (Allium sativum L.) are propagated through cloves, infection by virus or other pathogens may become severe problem if not using high quality seed bulbs every year resulting in the reduction of yield and bulb quality, In order to solve this problem, the establishment of virus-free bulb production and its supply system have been required because no chemicals were found to eliminate viruses from seed bulbs. This experiment was conducted to develop an effective production technique of high quality seed bulbs using shoot-tip culture. Over 90% of shoot-tips explanted on January L 1990 were survived at constant temperature of either 20, 24 or 28$^{\circ}C$, wheres 88% at alternate temperature (28/20$^{\circ}C$). The growth of shoot and root was most vigorous at constant 24$^{\circ}C$, and least at alternate temperature (28/20$^{\circ}C$) condition. When shoot-tips were explanted June 21 to August 1,1991, survival and growth of shoot-tips was most vigorous on MS medium supplymented with 0.1 mg/L NAA and 2 mg/L kinetin and least 1 mg/L Gh$_3$. The shoot-tips taken from the seed bulbs stored at 4$^{\circ}C$ for 15 to 60 days were placed on MS medium, shoot growth and in vitro bulblet formation increased slightly as affected by the increase of told treatment period at 4$^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.157-165
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• 1975

This experiment was carried out to evaluate the sanitary quality of commercially frozen sea foods. One hundred and sixteen samples in six different items from several refrigeration plant in Busan city were examined from March to December in 1974. In addition, the changes in bacterial density through the process from thawing, round or semifilleted frozen alaska pollack to the finishing as frozen fillet blocks were observed. To evaluate the sanitary quality, sanitary indicative bacteria such as total coliform, fecal coliform, fecal streptococci and enterococci as well as plate counts were determined. From the results, the median value of fecal coliform MPN was 20 per 100 grams of the samples and that of enterococci was 790. The median value of plate counts was $2.2\times10^4$ per gram. The plate counts were not correlated with the number of sanitary indicative bacteria. The results suggest that enterococci could be used advantageously in preference to coliform organisms as indicative bacteria for the evaluation of sanitary quality of frozen sea foods. The plate counts at $20^{\circ}C$ of the samples were 14 times higher than that at $35^{\circ}C$. Geometric mean of total coliform MPN was 310 and that of enterococci was 143. Bacterial density was reduced by fleering. Morethan 50 percent for total coliform MPN and $35^{\circ}C$ plate counts, and about 35 percent for enterococci MPN and $20^{\circ}C$ plate counts were reduced under the contact freezing unit which was generally operated at $-40^{\circ}C$. About fifty-five percent of the samples were negative in fecal coliform test and 10 percent of those were exceeded $1.0\times10^5$ per gram in $35^{\circ}C$ plate counts.

• The korean journal of orthodontics
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• v.27 no.3 s.62
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• pp.503-513
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• 1997

This study was undertaken to investigate the effect of oxygen tension on the activity and function of the cells derived from human periodontal ligament by measuring cell activity, total protein synthesis, collagen synthesis, $IL-1{\beta},\;IL-6,\;TNF-{\alpha}$ Human periodontal ligament fibroblasts were collected from premolars extracted for orthodontic treatment and incubated in the environment of $37^{\circ}C,\;5\%\;CO_2,\100\%$ humidity. After the fifth to sixth passage they were used for the experiment. Gaspack system to which $0.2{\mu}m$ Millipore filter was attached was connected to mixed-gas tanks. The mixed gases were composed of $10\%\;O_2,\;5\%\;CO_2,\;85\%\;N_2$ in hyoxic group or $90\%\;O_2,\;5\%\;CO_2,\;5\%\;N_2$ in hyperoxic group and $5\%\;CO_2,\;95\%$ air for control. After incubation in $37^{\circ}C$ for 2, 4, 6 days, cell activity was determined by tetrazolium(MTT) assay and total protein synthesis was assayed using sulforhodamine B(SRB). And measurement of 4-hydroxyproline was performed to assess collagen synthesis md $IL-1{\beta},\;IL-6,\;and\;TNF-{\alpha}$ were measured by enzymeimmunoassay. The results were as follows. 1. The cell activity and total protein synthesis in hypoxic group were a little higher than or almost the same with those in control group. 2. In hyperoxic group, the cell activity was lower than that in control group and total protein synthesis was decreased. 3. Collagen synthesis was significantly decreased initially in both hypoxic and hyperoxic group and increased nearly to the level of control group as the duration of cell incubation was longer 4. As a result of enzymeimmunoassay, the amount of cytokines was $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ in order. 5. $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ were increased more rapidly in both hypoxic and hyperoxic group than in control group as the duration of cell incubation was longer. 6. There were more $IL-6\;and\;TNF-{\alpha}$ in hyperoxic group than in control group after 6 days, and there were more $IL-6\;and\;TNF-{\alpha}$ after 6 days than after 2 or 4 days in hyperoxic group. These results suggested that oxygen tension might modulate the production of extracellular matrix and cytokines in the cells derived from human periodontal ligament.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.87-94
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• 2008

Porcine epidemic diarrhea virus (PEDV) is an infectious and highly contagious virus of swine. In order to develop the transgenic tobacco culture cells producing PEDV antigen protein, four vectors expressing PEDV spike protein (SP) gene under the control of a CaMV 35S promoter were constructed. Four fragments of the SP region of PEDV, SP1 (444 bp, 1487-1930 bp), SP2 (1.7 kb, 2300-3987 bp), SP3 (1.4 kb, 1559-2950 bp), and SP4 (2.6 kb, 9-2643 bp) were amplified by PCR and then C-MYC tag was fused to the end of each SP gene, respectively. These cassettes are inserted into the pCAMBIA2300 (named as 35S::SP1-M, 35S::SP2-M 35S::SP3-M, and 35S::SP4-M, respectively). Tobacco (cv. BY-2) cultured cells were transformed by co-cultivation with Agrobacterium tumefaciens harboring expression vector. We selected kanamycin-resistant calli and checked for the presence of the introduced SP gene using PCR, resulting 70% of them showed the foreign gene. We selected the lines with high-level expression of PEDV antigen protein based on dot blot analysis. Southern blot analysis confirmed that the PEDV SP gene was integrated into the genome of the tobacco cultured cells. Northern blot analysis showed that the introduced gene was highly expressed in transgenic cultured cells. Transgenic tobacco cultured cells-derived antigen induced immunogenicity in mice as determined by a plaque reduction neutralization assay. These results suggest that the vectors expressing PEDV spike protein gene in this study will be useful for the development of transgenic plants and cultured cells producing PEDV antigene protein.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.407-412
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• 2017

The aim of this study was to shorten the time required for subculture and bacterial identification and obtain a simple and rapid identification method for new test methods for bloodstream infections. The following results were obtained using a mass spectrometer. In Vitek 2, 208 (81.8%) cases were well-identified and 45 isolates were not identified in blood cultures. Among 208 cases, 146 (57.5%) were Gram positive bacteria and 108 (42.5%) were Gram negative bacteria. In total, 233 were identified to the species level and 21 were identified to the genus level. The identification error was found to be Propionibacterium acnes as Clostridium bifermentans. The accuracy of Enterobacteriaceae, glucose non-fermentative bacilli (GNFB), and staphylococci were 81/83 (97.6%), 12/15 (80.0%), and 72/85 (84.7%), respectively. The concordance rate of Vitek 2 and Vitek MS by the direct method was 81.8% and 45 isolates were not identified. Most of the unidentified bacteria were Gram positive bacteria (N=37). The Gram positive bacteria were streptococci (14), coagulase-negative staphylococci (CNS) (11), enterococci (3), Staphylococcus aureus (2), Micrococcus spp. (2), Bacillus spp. (2) and Actinomyces odontolyticus, Finegoldia magna, and Peptostreptococcus spp. The results reporting time was reduced to 24~72 hours compared to the conventional method. The rate of identification of the aerobic and anaerobic cultures was similar, but the use of an anaerobic culture did not require a dissolution process, which could shorten the sample preparation time. These results suggest that the method of direct identification in blood cultures is very useful for the treatment of patients. In further studies, it might be necessary to further improve the method for identifying streptococci and CNS, which were lacking in accuracy in this study.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.179-183
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• 2002

This study was carried out to investigate the influence of medium composition for in vitro mass propagation of Lycoris squamigera Max. After the disks of short stems, segments of leaf within bulb and scale were cultured on MS basal medium supplemented with various plant growth regulators, they were examined for the extent of callus formation, shoot and root regeneration. In the culture of stem disks, adventitious shoots were regenerated from the basal tissue of bulb scales, and combined medium of 1.0 mg/L 2,4-D or NAA＋2.0 mg/L BA or kinetin showed the the best response and 4∼6 shoots per explant formed. In the culture of leaf segments within bulbs, both MS medium supplemented with 1.0 mg/L NAA＋2.0 mg/L TDZ and with 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA were produced callus profusely on the base of leaf tissue and 3∼6 shoots were regenerated per explant. In the scale segments culture, calli were produced on the basal tissue on medium with 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA. The best result were shown on MS medium with 1.0 mg/L NAA＋2.0 mg/L TDZ, and 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA. Maximum number of regenerated shoots was up to 10∼12. Adventitious root formation from explants were formed profusely on MS medium with 1.0 mg/L NAA＋2.0 mg/L kinetin. The most desirable method for mass propagation of plantlets was the shoot regeneration from scale segments then subsequently subcultured on medium for rooting.

• Korean Journal of Veterinary Research
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• v.9 no.1
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• pp.23-62
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• 1969

Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

• Tuberculosis and Respiratory Diseases
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• v.57 no.1
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• pp.37-46
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• 2004

Background : Lung pericytes are important constituent cells of blood-air barrier in pulmonary microvasculature. These cells take part in the control of vascular contractility and permeability. In this study, it was hypothesized that change of lung pericytes might be attributable to pathologic change in microvasculature in acute lung injury. The purpose of this study was how hypoxia change proliferation and genetic expression in lung pericytes. Methods : From the lungs of several Sprague-Dawley rats, performed the primary culture of lung pericytes and subculture. Characteristics of lung pericytes were confirmed with stellate shape in light microscopy and immunocytochemistry. 2% concentration of oxygen and $200{\mu}M$ $CoCl_2$ were treated to cells. Tryphan blue method and reverse transcription-polymerase chain reaction were done. Results : 1. We established methodology for primary culture of lung pericytes. 2. Hypoxia inhibited cellular proliferation in pericytes. 3. Hypoxia could markedly induce vascular endothelial growth factor(VEGF) and smad-2. 4. Hypoxia-inducible factor-$1{\alpha}$(HIF-$1{\alpha}$) was also induced by 2% oxygen. Conclusion : Viability of lung pericytes are inhibited by hypoxia. Hypoxia can stimulate expression of hypoxia-responsive genes. Pericytic change may be contributed to dysfunction of alveolar-capillary barrier in various pulmonary disorders.