• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.5
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• pp.767-773
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• 1998

In this study, we have investigated the distributions and killing effects of marine bacteria that tend to kill the red tide microalgae, C. polykikoides in the area of Masan bay from June to October, 1996. To summarize, C. polykikoides killing bacteria were detected at $10^2$ to $10^3$ cells/ml of seawater samples during the survey period, and the bloom was observed in September by containing $4.8\times10^3$cells/ml. It appears however that the number of these bacteria is decreased ($2.0\times10^2$cells/ml) in October, A total of 110 strains were isolated from seawater samples and seawater filtrate (pore size, 0.8 $\mu$m)-containing mixed culture of C. polykikoides in which the mixed culture was grown in f/2 medium. As results we have successfully isolated Micrococcus sp. LG-1 which decreased to less than 10cells/ml within 6days and 5days sfter inoculation of Micrococcus sp. LG-1 into the la9 and logarithmic growth phases of C. polykrikoides respectively. Therefore, it appears that inoculation of Micrococcus sp. LG-1 against the logarithmic C. polykrikoides is more effective than the lag growth phase, (n addition, the killing effects were increased in accordance with bacterial cell densities inoculated in a dose dependent manner. Especially, the filtrate of kitling bacterium culture (nore size, 0.2 $\mu$m) revealed a dramatic effect in which C. polykrikoides were decreased to less than 10 cells/mf of culture within 1 hr, 1,5 hrs, 1,5 hrs, 3.5 hrs. and 5,5 hrs after inoculations of the culture filtrate with concentration of $30\%,\;20\%,\;10\%,\;5\%$ and $2.5\%$, respectively. Moreover Micrococcus sp. LG-1 showed a selective specificity against C. polykrikoides and any other killing effects of Micrococcus sp. LG-1 were not observed against Alexandrium tamarense, Prorocentrum micans, Scrippsiella trochoidea. ana Gymnodinium sanguineum.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.4
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• pp.289-296
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• 1990

The antimicrobial and antioxidant activities of grapefruit seed extract(GFSE), which was extracted with glycerine in the special schematic extraction apparatus, were investigated for handling and processing of fishery products. The effectivity of GFSE has been tried on sardine, mackerel and shrimp divided into six lots for each fishery product: control(no treatment) and five GFSE-treated samples. Samples were inoculated with Salmenella typhi, incubated for 24hrs at $30^{\circ}C$ in dextrose-tryptone broth medium and prepared for microbiological 8f chemical analysis and organoleptic assessment. The bacteriological analytical results with GFSE(250ppm) showed the reduction of $1.8\times10^6\to2.0\times10^4,\;1.9\times10^6\to1.8\times10^4$ and $1.6\times10^6\to2.7\times10^3$ in total bacterial count for sardine, mackerel and shrimp, respectively. The test results with GFSE(500ppm) showed a $100\%$ reduction of bacterial mackerel treated with GFSE(500ppm) was reduced to $1.1\times10^4$ and $9.0\times10^3$ respectively. Antioxidant effect of treatment with GFSE at 500ppm level for three products was significant. LSD test results on organoleptic parameter for the samples treated with various showed a significant influence on the appearance, odor and texture in which at concentration 500ppm level give the excellent scours compared to each control.

• Restorative Dentistry and Endodontics
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• v.31 no.6
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• pp.437-444
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• 2006

This study was aimed to elucidate the biological function of OD314 (Apin protein), which is related to ameloblast differentiation and amelogenesis. Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. We examined expression of the OD314 mRNA using in-situ hybridization during tooth development in mice. Expression of OD314 and several enamel matrix proteins were examined in the cultured ameloblast cell line up to 28 days by reverse transcription-polymerase chain reaction (RT-PCR) amplification. After inactivation and over-expression of the OD314 gene in ameloblast cell lines using U6 vectordriven RNA interference and CMV-OD314 construct, RT-PCR were performed to evaluate the effect of the OD314 during amelogenesis. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were more strongly expressed in ameloblast than odontoblast. 2. When ameloblast cells were cultured in the diffcrentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expressions of amelogenin and enamelin were gradually decreased according to the ameloblast differentiation. 3. Inactivation of OD314 by U6-OD314 siRNA construct down-regulated the expression of OD314, MMP-20, and tuftelin, whereas over-expression of OD314 by CMV-OD314 construct up-regulated the expression of OD314 and MMP-20 without change in tuftelin. These results suggest that OD314 is considered as an ameloblast-enriched gene and may play the important roles in ameloblast differentiation and mineralization.

• Journal of Korean Society of Forest Science
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• v.79 no.3
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• pp.302-308
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• 1990

This study was conducted to determine the effects of mycorrhizal inoculation and ortet ages on the rooting of Quercus acutissima cuttings. The cuttings taken from 12-week-, 2-year-, 4-year-, and 20-year-old plants were rooted in mid summer in the rooting medium (vermiculite 2 : peatmoss 1 by volume) with or without Pisolithus tinctorius (Pt) and Glomus sp. inocula and IBA under intermittent misting system in an open shed. The average percentages of rooting were about 82%, 49%, 29%, and 13% for cuttings taken from 12-week-, 2-year-. 4-year-, and 20-year-old seedlings, respectively. Pt inoculation, enhanced rooting of cuttings at all age classes, except 12-week-old seedlings, with the highest enhancement(22%) observed in cuttings taken from 20-year-old trees. The highest percentage of rooting in each age group eras 88.9% in 12-week-old seedlings treated with Pt plus 3.000ppm IBA, 75% in 2-Year-old plants with 1,000ppm IBA, 58.3% in 4-year-old plants with 3.000ppm IBA and 22% in 20-year-old plants. The addition of Glomus sp. fungus inoculum failed to enhance rooting. Pt mycorrhizal inoculation enhanced root dry weight, length, and diamter of adventitious roots at cuttings taken from 12-week- and 20-year-old trees, except the cuttings taken from 4-year-old seedlings. Rooted cuttings had more total nitrogen content in the leaves than unrooted cuttings, and the greater rooting response was associated with the higher phosphorus content in the leaves.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.619-624
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• 2011

Tea extract (TE) has been shown to have anti-tumor properties in a wide variety of experimental systems. We evaluated green tea extract (GTE) as a biochemical modulator for the antitumor activity of cisplatin and doxorubicin in the treatment of human lung cancer A549 cells. Cells were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and two antibiotics (100 units/mL penicillin and $100\;{\mu}g$/mL streptomycin). Two types of TE, epigallocatechin galate (EGCG) and GTE, were used in this experiment. The cells were seeded at $1{\times}10^4$ cells/well in the RPMI-1640 media with or without TE ($100\;{\mu}g$/mL) and then treated with different concentrations of doxorubicin ($0{\sim}14\;{\mu}g$/mL) or cisplatin ($0{\sim}35\;{\mu}g$/mL). After incubation in 5% $CO_2$ at $37^{\circ}C$ for 24 hr, cell viability was determined with a MTT assay. We used a Western blot to detect the influence of EGCG and GTE on the expression of p53 and caspase-3 genes in the A549 cells. A549 cell viability decreased to 15% with a $10\;{\mu}g$/mL concentration of cisplatin, and to 21% with a $8\;{\mu}g$/mL concentration of doxorubicin, as measured with the MTT assay. However, pre-treatment of the cells with EGCG ($100\;{\mu}g$/mL) or GTE ($100\;{\mu}g$/mL) resulted in decreased cell viability with $6\;{\mu}g$/mL of cisplatin and $4\;{\mu}g$/mL of doxorubicin. There was no apparent change in cell viability between EGCG or GTE administration in cisplatin- or doxorubicin-induced cytotoxicity in A549 cells. The levels of p53 and caspase-3 in the A549 cells increased with both EGCG and GTE treatment. We found that GTE could potentially affect cisplatin- or doxorubicin-induced cytotoxicity of A549 cells, which may be useful in the chemotreatment of cancer.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.171-178
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• 1999

The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF. And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF (Table 1). The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF. The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% in control, 8.77% in 10% FF and 20% in 40% FF (Table 2). On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively (Fig. 1, 2, 3). In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after $8{\sim}16$ cell stage.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

• Journal of Korean Society of Forest Science
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• v.38 no.1
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• pp.13-18
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• 1978

In this study, enzymatic hydrolysis of the holocellulose from Alnus hirsuta (Spach) Rupr. (8-14 yr's) was investigated using crude cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374. And conducted on the optimum condition of the treated substrate for saccharification. A strain of Trichoderma viride Pers. ex. Fr. SANK 16374 was found to be highly efficient for the cellulase productivity, especially in the submerged culture process. The culture medium used in this experiment was prepared from an extract of wheat bran consisting also of $KH_2PO_410$, $(NH_4)_2$ $SO_4$ 3, $NaNO_3$ 3, and $MgSO_4$ $7H_2O$ 0.5g/l. Cellulose powder (Toyo filter paper, 60 mesh) was found to be an importent factar for inducing the cellulase formation. And the cellulase produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate (Fig. 1) Reducing sugar was determined by the Dinitrosalicylic acid (DNS) method, using reagents prepared according to the method of Sumner (1925). The results obtained were summerized as follows; 1. The method of delignification were treated by the Peracetic acid (PA) method, according to the method of Toyama (1970). The yield of holocellulose were decreased in accordance with increasing concentration of Peracetic acid solution; delignification of Alnus hirsuta Rupr. with 20% Peracetic acid was satisfied for 48 hours and 40%~60% peracetic acid was satisfied for 24 hrs: 2. The substrate (holocellulose) was changed easely into fine powder with enzymatic hydrolysis and cellulase exhibits optimum activity on the reducing sugar formation from substrate at the range of 60-100 mesh. 3. The reducing sugar formation increased in accordance with increasing dry temperature on holocellulose substrate was found to be $190{\pm}5^{\circ}C$. 4. The optimal heat treated time of holocellulose substrate was found to be 45 min. for the reducing sugar formation showed the best products. The reducing sugar formation did not show statisticaly significent diflerences at 5% levels by heat treated time for 45 min. and 60 min.

• Journal of Bio-Environment Control
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• v.27 no.4
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• pp.407-414
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• 2018

This study was aimed to evaluate the effect of application method and concentration of gibberellin $A_3$ ($GA_3$) on the growth, runner production, and seedling quality of strawberry plants (Fragaria ${\times}$ ananassa Duch. cv. Maehyang) during nursery period. The mother plants of strawberry were transplanted in pot ($64{\times}27{\times}18cm$) filled with commercial growing medium on March 20, 2018. $GA_3$ concentration was applied as 0, 50, 100 or $200mg{\cdot}L^{-1}$ with spray or drench to 45 mL per plant at 4 weeks after transplanting, respectively. Nutrient solution was supplied with the EC $1.5dS{\cdot}m^{-1}$ after the transplanting and supplied 350 mL per pot twice a day (15 min per one time) after rooting. The growth characteristics of mother plants of strawberry were measured at 7 weeks after treatment, and growth characteristics of daughter plants of strawberry were measured at 10 weeks after treatment. Runner length and diameter of mother plant was the longest or thickest in the spray with $200mg{\cdot}L^{-1}$ than the other treatments, respectively. Soil-plant analysis development (SPAD) value of mother plant was the lowest in spray with $200mg{\cdot}L^{-1}$. However, leaf length, leaf width, and crown diameter showed no significant differences in all treatment among application method and concentration of $GA_3$. As the concentration of $GA_3$ increased, physiological disorder like stretchiness of crown occurred more. The physiological disorder was the most occurred in spray treatment with $200mg{\cdot}L^{-1}$, but drench treatment occurred less than spray treatment. The number of runners and daughter plants increased with increasing concentration of $GA_3$ regardless of application methods. In the growth characteristics of the daughter plants, leaf length and leaf width of first daughter plant, plant height, crown diameter, leaf area and SPAD value of second daughter plant, and plant height of third daughter plant were the significantly greatest in drench with $100mg{\cdot}L^{-1}$ treatment. This results indicate that growth and runner production of mother plants and growth of daughter plants of strawberry were the best achieved by drench application in the $100mg{\cdot}L^{-1}$ $GA_3$.

• Journal of Bio-Environment Control
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• v.27 no.4
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• pp.285-293
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• 2018

This study aimed to evaluate the effect of height suppression of cucumber and tomato plug seedlings as affected by mechanical stimulus using brushing as environment-friendly method. Cucumber (Cucumis sativus L. 'Joeunbaekdadagi') and tomato (Solanum lycopersicum L. 'Mini Chal') seeds were sown in 40-cell plug trays ($54{\times}27.5{\times}5cm$) filled with growing medium on Oct. 9, 2017. The cultivation environment in a venlo-type glasshouse was maintained as cultivation temperature range of $15-25^{\circ}C$ and the relative humidity of $50{\pm}10%$. Nontreatment and diniconazole ($7.5mg{\cdot}L^{-1}$) application at 15 days after sowing were used as the control. In addition, brushing treatments in cucumber and tomato were applied interval of 2, 4 or 6 hrs for 15 and 20 days, respectively. Plant height, hypocotyl length, and internode length were inhibited for cucumber and tomato in the diniconazole treatment than in the control. The leaf size was reduced, both cucumber and tomato, while the SPAD increased under the diniconazole treatment. However, stem diameter of cucumber was the thickest in the 2 hrs brushing interval treatment. Fresh weights of shoot and root were the significantly lowest in the diniconazole treatment. Application of brushing improved seedlings quality by promoting dry weights of shoot and root, and compactness of tomato seedlings. The chlorophyll fluorescence of tomato seedlings drastically decreased with 2 hrs treatment, indicating that mechanical stress by brushing treatment. The relative growth rate of tomato seedlings was significantly lower in the diniconazole treatment, but cucumber seedlings were not significantly different in all treatments. As a results, height suppression of cucumber and tomato seedlings was best achievement in the diniconazole treatment by the chemical as growth regulator. In an environment-friendly point of view, however, it is considered that 2 hrs brushing interval treatment can be the applicability for replacing the chemical methods in plug seedling growth of cucumber and tomato.