• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.293-303
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• 2007

Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.

• The Korean Journal of Mycology
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• v.41 no.3
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• pp.172-180
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• 2013

Grey mold infection rate in tomato was investigated with the inoculation of dead flowers on Botrytis selective media. The grey mold infection rate of flower after fruiting were higher in the order of after 45 days, after 25 days, and fruiting day with 100%, 87% and 65%, respectively. The number of infected flowers were increased with time increase after the flowering before fruiting. BSM (Botrytis selective medium) was used to check grey mold infection rate depending on the flowering stage and cultivar. Grey mold infection rate depending on the flowering stage was similar in all the beef-tomato cultivar as 1.5~5% at preflowering, 1.5~45% at flowering and 75~90% at fruiting. On the other hand, cherry tomato cultivar "KoKo" had lower infection rates of 0~3.5% at pre-flowering, 10~30% at flowering and 20~50% at fruiting. These resulted from the fact that beaf-tomato cultivar have much bigger flowers and larger amount of pollens compared to those of cherry tomato cultivar. The amounts of falling pollens of Botrytis spp. were checked for beaf-tomato cultivar and cherry tomato cultivar using BSTM. The amounts of falling pollens were increased as growth period was extended, and the amount of spores increased rapidly during the outbreak of grey mold. Twelve field trials in Buyeo and Iksan areas showed that Fluazinam, and Diethofencarb+Carbendazim were effective fungicides to control tomato grey mold, and these results were similar to those of field trials with BSTM. This is the first report of Fluazinam as a effective fungicide for the control of grey mold of tomato even though it has not been registered yet for the control of gray mold in tomato.

• Journal of the Korean Chemical Society
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• v.48 no.1
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• pp.28-32
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• 2004

The condensation reaction of (benzylene)triphenylphosphoranes with carbonyl compounds in supercritical carbon dioxide was examined. Reactions of (benzylene)phosphoranes (ca. 1 mmol) with several benzaldehydes in a supercritical carbon dioxide (80 $^{\circ}C$, 2,000 psi) containing THF entrainer (5%) in a 24 mL reactor proceed smoothly to yield olefination products in fairly good to excellant yields but slower, compared to reactions in a conventional THF solvent. Generally, phosphoranes that are not substituted with a nitro group show more (Z)-selective reactions with aromatic aldehydes under $scCO_2$ condition than in THF. The reaction of (benzylene)triphenylphosphoranes with 4-t-butylcyclohexanone gave the corresponding olefin compounds with a low conversion under both the supercritical carbon dioxide and the organic THF solvent. Our preliminary study showed the Wittig reaction carries out smoothly in supercritical carbon dioxide medium and also a possibile tunability of this reaction pathway by adding a entrainer. The results would be useful for devising a novel process for the environmentally friendly Wittig reaction.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.361-367
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• 2010

Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.1
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• pp.77-84
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• 1987

The population of actinomycetes in paddy soils, with hay compost and inorganic fertilizer had been applied respectively was investigated. Actinomycetes were isolated by using selective medium and the population densities of actinomycetes in paddy soil was examined. The population of actinomycetes were reached at a range from $2.1{\times}10^6$ to $7.4{\times}10^7$ per gram of the soil. The composition of actinomycetes flora changed considerably after hay compost applied. The significant positive correlations between the organic matter content in paddy soil and the actinomycetes populations were given at 1.38 to 2.69 level. According to the result of several morphological observation, similar strains isolated were classified into 21 groups. More detailed taxonomic characterization were carried out on the isolated strains. Therefore, 15 groups of Streptomyces and 6 groups of non-Streptomyces were classified into actinomycetes isolates, percentage of streptomyces and non-Streptomyces strains were 87.2% and 12.8% in the isolated 250 actinomycetes strains respectively. Streptomyces with sporophore of the spiral chain form accounted for 80% of all the Streptomyces isolates. Surface morphology of spores were determined with the electron microscope, three species have a spiny surface, and 13 strains have a smooth spore surface.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.327-335
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• 1996

Recently in spite of the interest on the regulation of intracellular $Mg^{2+}$ by neurotransmitters or drugs, the magnesium ion($Mg^{2+}$) regulation by ${\alpha}_1$-adrenoceptor stimulation has not been studied in the heart yet. To elucidate the regulation of ${\alpha}_1$-adrenoceptor stimulation-induced $Mg^{2+}$ release and the effects of ${\alpha}_1$-adrenoceptor stimulation on pathophysiological conditions, in this study we have evaluated the effects of phenylephrine, PMA, $H_7$. staurosporine, verapamil and lidocaine on $Mg^{2+}$ release in perfused guinea pig heart. During preperfusion exogenous $Mg^{2+}$ was added to the medium to give 1.2mM 15min before starting to addition of drugs, and then the infusion of exogenous $Mg^{2+}$ was stopped. $Mg^{2+}$ in the perfusate leaving the heart was measured by atomic absorption spectrophotometry. $Mg^{2+}$ free solution produced an increase in heart rate and phenylephrine elicited $Mg^{2+}$ release from the heart. $Mg^{2+}$ release by phenylephrine was abolished by combined treatment with prazosin. By contrast, cardiac $Mg^{2+}$ uptake induced by a protein kinase C(PKC) activator, PMA was abolished by a selective PKC inhibitor, staurosporine. And the phenylephrine-induced $Mg^{2+}$ release was not affected by the PKC inhibitor, $H_7$. When verapamil or lidocaine was added to perfusing solution, $Mg^{2+}$ release was potentiated by phenylephrine from perfused guinea pig heart. These results suggest that ${\alpha}_1$-adrenoceptor stimulation caused $Mg^{2+}$ release and that PKC is not involved in ${\alpha}_1$-adrenoceptor mediated $Mg^{2+}$ release from perfused guinea pig heart. Under pathophysiological conditions, the $Mg^{2+}$ alteration by ${\alpha}_1$-adrenoceptor stimulation is considerable.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.125-131
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• 1986

In order to obtain the optimum conditions for intact yeast cell transformation in the various yeast host-vector systems, 3 yeast plasmid vectors, YRp7, YEpl3 and YIp5 were introduced into 5 yeast hosts, Saccaromyces cervisiae Dl3-1A, DKD-5D, DBY-746, MC-16 and S2022D with various transformation conditions, and plasmid stabilities in all the transformants were also observed. The highest transformation frequencies in all the host-vector system were obtained in the 16 hour Cultured cell (5.4 $\times$ 10$^6$ - 2.4 $\times$ 10$^8$cells/$m{\ell}$) treated with 0.1-0.2 M lithium chloride in 0.1 M tris-HCl (pH 7.6), 35% polyethylene glycol 4000, and heat-shocked at 42$^{\circ}C$ for 5 minutes after 60 minutes of induction. The intact cell transformation got more transformation frequency in DKD-5D (YRp7) and DBY-746 (YEpl3) than protoplast transformation, but reverse tendency was observed in DKD-5D (YEp13) and Dl3-lA (YRp7). The transformants, D13-1A (YRp7) and DKD-5D (YRp7) were very unstable in selective medium, with 80 to 85% of the transformants losing the plasmid after 70 generations, but the transformants, DKD-5D (YEpl3) and DBY-746 (YEpl3) were quite stable, with 35% of the transformants losing the plasmid.

• Korean Journal of Environmental Biology
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• v.24 no.4
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• pp.307-313
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• 2006

Seasonal variation of marine bacterial community was analyzed in the surface sea water collected from one of the stations locating at Tongyeoung coastal area, Korea. The results obtained by the culture method through identification with the VITEK Microbe ID system after pure culture in the selective medium were compared with those obtained by the DGGE based 16S rRNA PCR method. The composition of the marine bacterial community in the sea water samples harvested in September, 2004, November, 2004, January, 2005, May, 2005 and August, 2005 determined by the culture method showed 5, 5, 4, 6, and 10 strains respectively. Pseudomonas fluorescens and Acinetobacter lwoffii were detected in all seasons. The other strains were identified to be Pseudomonas stutzeri, Sphingomonas paucimobilis, Burkholderia mallei and Chryseobacterium indologenes. In contrast, the 16S rRNA PCR-DGGE method detected 10, 11, 6, 9 and 13 populations respectively in the same sea water samples and the strains were identified to be Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticue, Sphingomonas paucimobilis and Rugeria algocolus. This results indicated that the DGGE based 16S rRNA PCR method was more efficient than the culture method for the grasp of the characteristics of the marine bacterial community.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.465-470
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• 2016

Sample preparation methods were evaluated for effectiveness in detecting foodborne pathogens from sprout seeds. The methods included: Rinse.-Test portions were rinsed with 0.1% peptone water, and the pellet after centrifugation was inoculated into pre-enrichment media; and Sprouting.-Seed samples were sprouted before pre-enrichment and sprouted seeds were inoculated into pre-enrichment media. In rinse method, E. coli was isolated from 13 of 280 sample units. In sprouting method, E. coli was isolated from 12 of 135 sample units. E. coli O157:H7, Salmonella spp., and L. monocytogenes were not detected in any of the samples. In the trials for recovering Salmonella enterica from artificially contaminated alfalfa seeds, the soak, rinse, and sprouting methods were evaluated. The detection rates of S. enterica were statistically different according to the amount of the sample tested and selective medium type (P < 0.05).