• Journal of the Korean Applied Science and Technology
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• v.29 no.1
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• pp.71-80
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• 2012

Vinca alkaloids produced from Catharanthus roseus are one of the most important natural product drugs in treatments of human cancers. These anticancer drugs are derived from coupling of the two monomeric indole alkaloids, catharanthine and vindoline. In order to investigate vindoline biosynthesis, tabersonine-$CD_3$ 1a is synthesized to use as a deuterium labeled precursor, which is distinguished clearly from the natural counterpart. We show that these deuterium labeled tabersonine 1a are successfully incorporated into the vindoline biosynthetic pathway to yield three deuterated vindoline intermediates. 16-Hydroxytabersonine-$CD_3$ (m/z 356) 2a, 16-Methoxytabersonine-$CD_3$ (m/z 370) 3a, 16-Methoxy-2,3-dihydro-3-hydroxytabersonine-$CD_3$ (m/z 388) 4a are produced from the cell suspension culture measured by UPLC/MS at 5 and 13 days after feeding tabersonine. The conversion rates from 1a to 2a and 2a to 3a are fast, whereas that from 3a to 4a is much slower. This indicates that the rate determining step among the first three vindoline biosynthesis is the last step. As a result of the slow conversion rate from 3a to 4a, the accumulation level of 16-Methoxytabersonine-$CD_3$ 3a is significantly increased up to 13 days. The accumulation ratio among 2a, 3a and 4a is 1, 2 and 0.1 at 5 days. However, the peaks of desacetoxyvindoline-$CD_3$ 5a, deacetylvindoline-$CD_3$ 6a and vindoline-$CD_3$ 7a are not found from the cell extracts even after 13 days of incubation which may indicate no presence of their corresponding enzymes.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.163-169
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• 2001

OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P ＜ 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

• Journal of Food Hygiene and Safety
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• v.28 no.4
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• pp.349-353
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• 2013

This study was performed to evaluate the bactericidal efficacy of Fumagari OPP$^{(R)}$, fumigation disinfectant, containing 20% ortho-phenylphenol against Staphylococcus aureus (S. aureus). In this research, efficacy test of fumigant against S. aureus was carried out according to French standard NF T 72-281. S. aureus working culture suspension number (N value), all of the colony numbers on the carriers exposed with the fumigant (n1, n2, and n3), the number of bacterial test suspentions by pour plate method (N1), the number of bacterial test suspentions by filter membrane method (N2) and the mean number of bacteria recovered on the control-carriers (T value) were obtained from the preliminary test. In addition, the reduction number of S. aureus exposed with the fumigant (d value) was calculated using T value, the mean number of bacteria in recovery solution (n'1) and the mean number of bacteria on carriers plated in agar (n'2). N value was $4.0{\times}10^8$ CFU/mL, and n1, n2, and n3 were higher than 0.5N1, 0.5N2 and 0.5N1, respectively. Additionally, T value was $3.4{\times}10^6$ CFU/mL. In the bactericidal effect of the fumigant, the d value was 6.43 logCFU/mL. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. The results indicated that Fumagari OPP$^{(R)}$ had an effective bactericidal activity against S. aureus, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with pathogenic bacteria.

• The Journal of Korean Academy of Prosthodontics
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• v.50 no.2
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• pp.106-111
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• 2012

Purpose: The purpose of this study was to compare the effect of 3 chairside polishing methods and laboratory polishing methods on surface roughness and $C.$ $albicans$ adhesion of polyamide denture base. Materials and methods: Using contact profilometer, the surface of polyamide specimens ($25{\times}15{\times}2mm$) was studied after conventional polishing without finishing and after chiarside polishing with 2 chiarside polishing kits and chairside-pumice polishing following finishing with tungsten carbide bur. To evaluate the adhesion of $C.$ $albicans$, $C.$ $albicans$ suspension was overlayed on the test specimen. And the specimens were incubated for 2 hours. Imprint culture method was achieved and counted the colony on the agar plate. Polished polyamide were evaluated using a scanning electron microscope. The statistics were conducted using one-way ANOVA and in case of difference, Scheffe test and Tamhane's T2 test were used. Results: Surface roughness (Ra) of surfaces polished with 2 chairside polishing kits had higher than conventional polishing and pumice polishing. The highest roughness value was $0.32{\pm}0.10{\mu}m$, and the lowest was $0.02{\pm}0.00{\mu}m$. The adhesion of $C.$ $albicans$ on the specimens polished with chairside polishing group and pumice polishing group were increased than conventional polishing group ($P$<.01). Conclusion: Conventional laboratory polishing was found to produce the smoothest surface and the lowest adhesion of $C.$ $albicans$. Two groups polished with Chairside polishing kits were similar with respect to surface roughness. Surface of the specimen polished with pumice is significantly smoother than 2 chairside polishing groups, but the result of $C.$ $albicans$ adhesion is that group polished with pumice was similar with 2 chairside polishing groups ($P$>.01).

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.311-318
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• 1997

Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

• Research in Plant Disease
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• v.21 no.2
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• pp.58-66
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• 2015

This study was conducted to evaluate control efficacy of a fermented food 'Cheonggukjang' against cucumber powdery mildew caused by Sphaerotheca fuliginea in greenhouse. Sterilized Daepung beans were inoculated with the rice straw as natural inoculum and then incubated for 72 hrs at $42^{\circ}C$ in the household cheonggkjang maker. After 72 hrs of cheonggukjang fermentation, white zymogens were grown on the surface of a sterile Daepung beans. The pH of the 72 hrs fermented soybean was not significantly changed and electrical conductivity was found to increase by about 2 times than before fermentation. The population density of soybean zymogen showed a peak of growth at 60 hrs after fermentation and the concentration of zymogen was $8.2{\times}10^7cfu/ml$. Soybean zymogen form of the colony was divided into three kinds of bacteria and a white and a large colony (WL) was predominant bacteria among those up to 60 hrs of fermentation. To control the cucumber powdery mildew, diluted solutions of cheonggukjang was applied from 6.0% to 30.0% on cucumber leaves and they showed injury symptoms on cucumber leaves in more than 15% of them. However, more than 6.0% diluted cheonggukjang solutions showed more than 77.8% control effect of cucumber powdery mildew at 15 days after treatment. The fermented bacteria of Chenggukjang were well established in the cucumber leaf area at 15 days after treatment. The antifungal activity of 10% diluted cheonggukjang solutions was excellent for four species of plant fungal pathogens, Colletotrichum gloeosporioides, Sclerotinia cepivorum, Rhizoctonia sloani and Phytophthora capsici in the dual culture test. Results indicated that foliar application of Cheonggukjang solution could be used for the control of powdery mildews occurring on organically cultivated cucumber.

• Journal of Food Hygiene and Safety
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• v.29 no.1
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• pp.60-66
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• 2014

This test was performed to evaluate the bactericidal efficacy of a fumigation disinfectant containing 20% ortho-phenylphenol against Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus hirae (E. hirae). In preliminary tests, P. aeruginosa and E. hirae working culture suspension number (N value) were $2.8{\times}10^8$ and $4.0{\times}10^8CFU/mL$, respectively. And all the colony numbers on the carriers exposed to the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of bacterial test suspentions by pour plate method), 0.5N2 (the number of bacterial test suspentions by filter membrane method) and 0.5N1, respectively. In addition, the mean number of P. aeruginosa and E. hirae recovered on the control-carriers (T value) was $2.8{\times}10^8$ and $3.4{\times}10^6CFU/mL$, respectively. In the bactericidal effect of the fumigant, the reduction number of $2.8{\times}10^8$ (d value) was 6.46 and 5.19 logCFU/mL, respectively. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. With the results from this study, the fumigation disinfectant containing 20% ortho-phenylphenol has an effective bactericidal activity, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with the pathogenic bacteria.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.447-453
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• 2007

With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.379-387
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• 2013

In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.