• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.6
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• pp.812-813
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• 1995

Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.1
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• pp.10-18
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• 2011

Purpose: The purpose of this study is to find out the effects of bisphosphonates (BPs) on the proliferation and the alkaline phosphatase (ALP) activity of human bone marrow derived mesenchymal stem cells (hMSCs), and thus state its correlation with bisphosphonate related osteonecrosis of the jaw (BRONJ). Methods: hMSCs was obtained by collecting and culturing cancellous bone fragments from a patient undergoing iliac bone graft. Alendronate (Aln) and Pamidronate (Pam), Ibandronate (Ibn) were added to the culture media in the concentration from $10^{-3}$ M to $10^{-11}$ M and cell toxicity, viability were measured. For ALP activity evaluation, Aln and Pam were added to the culture media in the concentration from $5{\times}10^{-7}$ M to $1{\times}10^{-8}$ M and were cultured for 1 week, 2 weeks and 3 weeks. ALP activity data were standardized using protein assay. Control groups were prepared for each examination. Results: Aln, Pam and Ibn all failed to increase the proliferation of hMSCs. With 1 week, 2 weeks of $5{\times}10^{-8}$M of Aln treatment, the ALP activity increased. Pam treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and$1{\times}10^{-8}$M. Also Ibn treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and $1{\times}10^{-8}$ M. Conclusion: It is considered that BPs are not capable of improving the proliferation of hMSCs. Also, after a transient increase in the ALP activity with the lower concentration of BPs, the activity decreased again. Therefore, in patients on long-term medication of BPs, the proliferation and osteoblast differentiation of hMSCs are restrained, and thus delayed wound healing and increase in BRONJ complications may occur.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.1
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• pp.25-32
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• 2003

S. mutans is the most important causative bacteria of dental caries among the oral bacteria. S. salivarius is a normal inhabitant in the human oral cavity. Nine strains of S. salivarius in this study were isolated from the oral cavities of children and identified, and their effect on S. mutans and S. oralis was studied. 1. The mean weight of produced artificial plaque on the wires in the beaker was 204.9 mg in the culture of S. mutans only, whereas being reduced to 1.9 mg through 20.6mg in the combined culture of S. mutans and each S. salivarius isolate (p<0.05). The viable cell didn't show the difference between them after culturing. 2. When S. mutans was cultured in the media containing culture supernatant of each S. salivarius isolate in M17 broth, the mean weight of produced artificial plaque was 117.1 mg on the wires, whereas being 47.7 mg in the media containing culture supernatant of each S. salivarius isolate in M17 broth containing 5% sucrose. 3. The polymer produced by S. salivarius isolates was on the thin layer chromatography. 4. Inulin and levan didn't inhibit the formation of artificial plaque by S. mutans in the beaker test. These results suggested that fructan-producing S. salivarius isolates inhibited the formation of artificial plaque by S. mutans.

• The Korean Journal of Mycology
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• v.43 no.4
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• pp.239-246
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• 2015

Two fungal strains were isolated from rods of Quercus sp. (NAAS02335) and Pinus densiflora (NAAS05299) in Korea. These strains were identified as Gyrodontium sacchari by their morphological and mycological characteristics. The optimal growth temperature of NAAS02335 and NAAS05299 are $25^{\circ}C$ and $30^{\circ}C$, respectively. Production of cellulase, xylanase, and ligninase was tested on agar media supplemented dyes or substrates. Production of cellulase and xylanase of NAAS05299 was higher than those of NAAS02335, however ligninase activity of NAAS02335 was higher than that of NAAS05299. The activities of cellulase, xylanase, and amylase of strain NAAS05299 were estimated at 6.7~10.2 times higher than that of NAAS02335. Laccase activity was only estimated by strain NAAS02335. The lignocellulytic enzymes are induced by substrates such as rice straw, wooden chips of pine, oak, and poplar. The NAAS05299 was able to degrade filter paper completely after 4 weeks of culturing in liquid media containing a piece of filter paper at $28^{\circ}C$ with continuous shaking. NAAS05299 was able to degrade rice straw, pine chips, and oak chips after 4 months in solid culture, however NAAS02335 decomposed only rice straw among tested 4 kinds of biomass.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.39-46
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• 2000

Objective: A number of studies to improve in vitro culture conditions have been tried over past ten years by using co-culture system with helper somatic cells. However, the mechanism of coculture is poorly understood. This study was designed to understand the mechanism for the mode of actual action of co-culture using co-culture system of ICR strain's 1-cell embryos with human oviduct epithelial cells by examining the effect of conditioned medium and contactless coculture using a cell culture insert on the embryo development and by measuring the level of superoxide anion from conditioned medium after co-culture. Methods: ICR strain's zygote embryos were cultured in medium alone (control), coculture, conditioned medium, or contactless coculture system for 6 days. Conditioned media (CM) were prepared as following 5 groups. All CM were collected after culturing oviduct cells for 2 days. CM-1 was stored at $-20^{\circ}C$ until use, and CM-2 was prepared just before use as a culture medium. CM-3 was cocultured with embryos and retrieved just before use. CM-4 and CM-5 were derives from the microfilteration of CM-2 and CM-3, respectively, using Microcon-10 (10 kDa molecular weight cut-off). The percentage of the embryos developed to hatched blastocyst stage and the level of superoxide anion in supernatant from medium alone culture (control), coculture, and contactless coculture were measured. Results: The rates of embryo development to the hatched blastocyst stage were significantly higher in coculture (43%) than in control (0%) (p<0.05). The CM-1 group had no embryo development since 2-cell embryonic stage, whereas the CM-2, CM-3, CM-4 and CM-5 groups had the improved development to 4 or 8 cell embryo stage, but the similar rate of development to hatched blastocyst compared to control. The effect of coculture on embryo development was disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in coculture group compared to control. Conclusion: It is concluded that the present coculture system overcomes the 2-cell block in vitro and improves the embryo development. This beneficial effect may be due to the direct cell-cell contact between embryo and helper cells or the removal of deleterious components from medium rather than the embryotrophic factors.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.2
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• pp.45-56
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• 2001

Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

• Journal of Korean Society of Forest Science
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• v.79 no.2
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• pp.109-114
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• 1990

The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.233-239
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• 1997

Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).

• Korean Journal of Plant Resources
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• v.14 no.3
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• pp.196-205
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• 2001

The results of examining cultivation of tlyster mushroom using NSM(Non Sterilized Media) for determining the condition of artificially culturing oyster mushroom(Pleurotus ostreatus) are as follows. Mycelial growth an[1 density of oyster mushroom. were the highest in the medium of waste cotton(spinning) : corn cob(80 : 20, V/V) followed by the order of rice bran, beet pulp. Expecially, mycelial growth and density of oyster mushroom is the lowest at the mixture rate of 80% waste cotton(spinning) : 10% beet pulp. Mycelial growth and density of oyster mushroom. were the highest in the medium of cotton seed hull and beet pulp mixture followed by the order of rice bran, corn cob. Expecially, mycelial growth and density of oyster mushroom is the lowest at the mixture rate of 80% cotton seed hull : above 20% rice bran. Mycelial growth and density of oyster mushroom were the highest in the medium rate of 70% waste cotton(spinning), 10% corn cob and 10% beet pulp(V/V). Mycelial growth and density of oyster mushroom were the highest in the medium rate of 70% cotton seed hull , 10% corn cob and 10% beet pulp(V/V). Optimal concentration of NSM for the mycelial growth and density of oyster mushroom were shown to be 500 times concentration. Optimal water contents for the mycelial growth and density of NSM was 70%.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.743-760
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• 2014

This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.