• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.189-195
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• 1999

This study was carried out to clarify several factors affecting embryogenesis from anther culture of nine cultivars in Brassica oleracea L. var. italica and to investigate the characteristics of plants derived from anther culture. Androgenesis from anther culture was elevated on the B5 medium supplemented with 0.1mg/L NAA, 0.1mg/L 2.4-D and 10% sucrose. Embryo production in liquid medium was five-fold higher than solid medium. High temperature treatment at 35$^{\circ}C$ for one day before transfer to culture room maintained at $25^{\circ}C$ had effective to induce embryogenesis of cultured anthers but extended treatment at 35$^{\circ}C$ decreased significantly the percent of embryogenesis. Frequency of embryogenesis from cultured anthers exhibited significant difference from 2.8% in 'Green Valiant' to 21% in 'Haisi' as affected by genotypes. Percent of spontaneously dihaploid among regenerated plants from anther culture was ranged from 62 to 74% as affected by the genotypes. Characteristic in relation to plant height, number of leaves and branches, and size of head from anther-derived plants showed differential variation in 'Rokguray' and 'Haisi'. Among these charaters obtained from two cultivars, five lines were selected for early maturity, long plant height and large head. Selected lines were used as breeding meterials for F$_1$ hybrid.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.98-109
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• 2016

The study was aimed to investigate the mixing ratio of kiwi and persimmon juices for the production of good quality wine by Saccharomyces cerevisiae Y28. Firstly, the optimum condition of rapidase treatment for the kiwi and persimmon juices was established, thereafter various mixing ratio (10:0, 9:1, 8:2, 7:3, 6:4, 5:5) of kiwi and persimmon was investigated regarding physiochemical properties and flavor compounds of wine. As the result, the optimum conditions were obtained as 0.3% rapidase for 1 h in kiwi and 0.3% rapidase for 3 h in persimmon. According to higher ration of persimmon, the pH of wines increased from 3.69 to 3.77, while the acidity of wines decreased from 2.07% to 1.51% at 14 days fermentation. The ranges of brix and reducing sugar in wines were decreased which ranges around 9.6 to 8.8 and 6.07 to 6.90 g/L, respectively, after fermentation. Major organic acid in wines were identified as tartaric acid, malic acid, and citric acid. A small amount of free sugar such as sucrose and glucose were detected in wines, but fructose was completely absent. The soluble phenolic contents were decreased that ranges around 1.00 to 1.25 g/L, in contrast, browning degree were increased ranges around 0.212 to 0.412 after fermentation. The major flavor components were identified as ethyl acetate and hydrazine, and 1,1-dimethyl. Importantly, phenylethyl alcohol was detected from the all wines that have a typical rose like flavor. But sensory test results and preference of kiwi-persimmon (7:3) mixing wine was better than the other wines.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.67-74
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• 2000

Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.379-387
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• 2017

Various sizes (0.2 ~ 1.2 mm) and developmental stages (referred to as Stage 1 ~ 3) of apical and lateral meristems were excised, together or separately, directly from dormant buds of apple 'Hongro'. They were mixed infected by Apple scar skin viroid (ASSVd), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV), which are major viruses attacking apples. A total of 31 callus lines (> 10 mm in diameter) were obtained by culturing the explants on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA), and they were subjected to RT-PCR analysis for virus detection. A high rate of virus elimination (expressed as the percentage of calli that did not amplify during RT-PCR, i.e., RT-PCR negative calli per total number of calli obtained) was achieved for ACLSV (100%), ASSVd (93.7%), and ASPV (93.7%), whereas it was only 25.8% for ASGV. ASPV was detected in the presence of 2 ~ 3 bracts. Simultaneous virus elimination of ASSVd, ASPV, ACLSV, and ASGV occurred during the meristem culture, in which the early stages of the dormant buds (Stage 1) were used, because ASGV was mostly eliminated during that stage. The results of the present study will be valuable for the production of virus-free apple trees.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.59-64
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• 1998

This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).

• Journal of Applied Biological Chemistry
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• v.54 no.2
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• pp.126-134
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• 2011

In this study, the characteristics of alcohol fermentation using ripe Daebong persimmon juice were studied in static fermentation condition by Saccharomycess cerevisiae CS02 in an effort to develop new types of functional wine. Attempts were made to modify the ripe Daebong persimmon juice in order to find suitable conditions for alcohol fermentation. The modified ripe Daebong persimmon juice that was most suitable for alcohol fermentation contained $24^{\circ}brix$ of sugar supplemented with sucrose as a carbon source and 0.5 g/L of $(NH_4)_2HPO_4$ as a nitrogen source. After 9 days of fermentation at $25^{\circ}C$, $12.2{\pm}0.02%$ of alcohol was produced from the modified juice and its pH markedly decreased to $3.97{\pm}0.02$. The wine contained free sugar such as fructose ($0.12{\pm}0.02$ g/L), some organic acids such as malic acid ($35.92{\pm}0.24$ g/L), succinic acid ($8.12{\pm}0.03$ g/L), oxalic acid ($22.14{\pm}0.11$ g/L), and citric acid ($13.63{\pm}0.08$ g/L), as well as some flavanols and phenolic acids such as catechin gallate ($38.99{\pm}0.32$ mg/L), epicatechin gallate ($110.21{\pm}0.16$ mg/L), gallic acid ($163.88{\pm}1.11$ mg/L), epigallocatechin ($15.97{\pm}0.18$ mg/L), and tannic acid ($13.36{\pm}0.02$ mg/L). In addition, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (84.25%) and $ABTS^{\cdot+}$ radical (99.65%) scavenging activities were increased significantly with a corresponding increased in the organic acid and phenolic acid contents, but decreased in the flavonoids.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.307-317
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• 1998

This study was to investigate the vitrification method for cryopreservation technique of bovine in vitro fertilization(IVF) embryos. The morphological appearance and viability following vitrification of IVF bovine blastocysts and expanded blastocysts were examined. Embryos obtained 6, 7, 8 or 9 days after IVF were vitrified in both 40% ethylene glycoI(EG) plus 0.3M trehalose and 20% polyvinylpyrrolidone(PVP) in DPBS(ETP, Exp. 1) and ETP solution added 0.375M dextrose (ETPD, Exp. 2). The viability of Days 6, 7, 8 and 9 vitrified /thawed embryos at 24∼48 h culture after thawing was 11.9%, 19.8%, 23.4% and 15.3% in ETP(Exp. 1), and 34.6%, 54.5%, 37.9% and 13.0% in ETPD(Exp. 2), respectively. The viability of vitrified embryos produced from the culture days after IVF did not differ in Exp. 1, but significantly differ in Exp. 2(P＜0.05). The viability of blastocysts and expanded blastocysts significantly differed(P＜0.05) in 15.2% and 23.3%(Exp. 1), and 25.0% and 45.8%(Exp. 2). The result of Exp. 1 was similar to that of Exp. 2 on the viability of embryo according to developmental stages, but ETP solution plus sugar(dextrose) was increased the viability of vitrified embryos. In Experiment 3, The viability of vitrified embryos was not different between 12% and 20% PVP concentrations in ETP solution according to culture days or developmental stages. To investigate the effect of addition of sugar, two type of carbohydrates and a mixture of cryoprotectants for vitrification on the survival of bovine IVF embryos, bovine Days 7 to 9 blastocysts and expanded blastocysts were cryopreserved in either 20% glycerol plus 20% EG, 0.375M sucrose and 0.375M dextrose (GESD, Exp. 4) or 20% glycerol plus 20% EG, 0.3M trehalose and 20% PVP(GETP, Exp. 5) in DPBS. Survival rates of Day 7, 8 and 9 embryos at 24∼48h culture after thawing were 71.4%, 94.6% and 40.5% in GESD, and 59.5%, 81.5% and 62.5% in GETP, respectively. Hatching rates of Day 7, 8 and 9 embryos after thawing were 28.6%, 35.1% and 16.2% in GESD, and 27.0%, 33.3% and 18.8% in GETP, respectively. These results indicates that a mixture of cryoprotectants(glycerol and EG) and addition of sugar can improve the survival rates of the bovine IVF embryos(Day 7 or 8) vitrified, and the expanded blastocyst embryos are more suitable for vitrification than early blastocysts stage.

• Journal of the Korean Applied Science and Technology
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• v.37 no.3
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• pp.484-495
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• 2020

This study is a mixed surfactant (MimicLipid-MSM1000) that forms the same structure as that of the stratum corneum, sucrose distearate, polyglyceryl-2 dioleate, fermented squalane, ergosterol, 10-hydroxystearic acid, mixture consisting of was synthesized. When using 2~5 wt% of this mixed surfactant, it was possible to make an artificial skin mimetic that forms a multi-layer lamellar structure of 5~30 layers. An emulsion was prepared using this mixed surfactant, and a multi-layered lamellar phase was formed and analyzed mechanically. The appearance of this surfactant was a light brown hard wax, the hydrophilic lipophilic balance (HLB) was 12.53, the critical parameter value was 0.987, and the acid value was 0.13. Stability according to pH change was also stable in acidic (3.8), neutral (7.2) and alkaline (10.8). The particle size of the liquid crystal was found to be the most stable maltese cross lamellar crystalline droplet at 5~25mm. The size of the emulsified particles according to the change in the speed of the homo agitator is 2500 rpm (17.9mm±2.6mm), 3500rpm (12.5mm±2.1mm), 4500rpm (6.2mm±1.8mm) particles were formed. Liquid crystal forming particles were observed through a polarization microscope, and the formation structure of the liquid crystal was precisely analyzed with a scanning electron microscope (cryo-SEM). As an application field, it is expected that it will be widely applicable to the development of various prescriptions, such as various skin care cosmetics, makeup care cosmetics, and scalp protection cosmetics, by using a skin-mimicking surfactant.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.118-123
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• 1998

Lasiodiplodia theobrorme is a pathogen of Java black rot on sweet potato. This fungus infects the tuberous root during storage after harvest. Invasion of the fungus results in the expansion of necrosis sites into the tuberous roots. The resultant necrotic symptom of the tissue is also induced by application of acetone extract of the fungus growing on potato sucrose agar (PSA) culture. The active principles to induce the necrosis are purified from the acetone extract as follows. After evaporation of hexane-benzene-EtOAc (1:2:1, v/v/v) the extract was fractioned on silica gel column, using a solvent gradient system from n-hexane to EtOAc and then to MeOH. The active fractions were purified with HPLC on Nucleosil 50-5 column by eluting n-hexane to EtOAc. Their structures are established by using spectroscopic techniques and synthesis to 4-hydroxymellein and mellein, respectively. Application of small amount of these compounds induce expansion of the necrotic symptom into the tissue and accumulated ipomeamarone. Conclusively, these compounds acted as phytotoxins (inducing necrosis) and as elicitors (eliciting the phytoalexin).

• Journal of Korean Society of Forest Science
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• v.98 no.2
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• pp.142-147
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• 2009

Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.