• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.161-172
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• 1991

This study was conducted to research the endocrine mechanisms of postpartum anestrus and determine if the morphology of the CL could be related to function in Korean native goats. At parturition 48 goats were assigned to a nonsuckled group and a suckled group. Serum concentrations of luteinizing hormone(LH), follicle stimulating hormone(FSH), prolactin(PRL), estradiol-17$\beta$(E2) and cortisol were measured at various times after parturitionin the goats. The corpora lutea of pregnancy were examined by a light microscope on the 6th hour and the first, 3rd, 10th, 11th, and 21st days after parturition. The results were summarized as follows : Mean serum LH concentrations were lower after parturition in all treatments and increased gradually with the intervals after parturition(P<0.01). These values did not differ between groups. The levels of serum FSH were lower after parturition and tended to increase gradually between 2 and 21 days. The levels of serum FSH are not significantly different between the groups of goats. Two days after kidding mean levels of serum PRL began to fall in nonsuckling goats but increased in suckling goats. During 3 weeks serum PRL concentrations were different between nonsuckling and suckling goats(P<0.01). Three days after parturition the levels of serum E2 decreased in all treatments. From parturition to day 21 serum E2 concentrations were greater in nonsuckling than in suckling goats(P<0.01). At the sixth hour after parturition the structure of the CL was well preserved. At days 1 and 3 the blood vessels were sparcely distributed, whereas, at days 1 and 3 the blood vessels were sparcely distributed, whereas, at days 10, 11 and 21 tortuous larger vessels with thick walls were observed on the luteal tissue. At days 1, 3, 10, 11 and 21 after parturition the CL of pregnancy showed degeneration and the proportion of tissue occupied by intercelluar substances increased at days 21 postpartum. In conclusion, the present study has shown that regression of the CL of pregnancy is accelerated in the period after parturition and effectively completed within three weeks postpartum.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.173-177
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• 2000

연구목적: 한국여성의 다낭성 난포증후군 환자에서 황체형성 호르몬 exon 2 유전자의 변이를 탐색하여 이들 변이와 질환과의 관련성 여부를 밝히고자 하였다. 연구재료 및 방법: 21명의 다낭성 난포증후군 환자를 대상으로 황체형성 호르몬 exon 2(Trp8Arg;TGG to CGG and Ile15Thr; ATC to ACC)의 변이를 탐색하였다. 혈액에서 Genomic DNA를 추출하여 PCR로 증폭한 후 RFLP 방법으로 변이형을 구분하였다. 결과: 황체형성 호르몬 exon 2의 변이형이 다낭성 난포증후군 환자에서 28.6%로 이미 조사된 바 있는 대조군의 16.7% 보다 약간 높게 나타났으나 통계적으로 유의한 차이는 없었다(p>0.05). 결론: 황체형성 호르몬 exon 2의 변이가 한국인의 다낭성 난포증후군 발병과 관련이 있는지를 밝히기 위해 더 많은 개체에 대한 연구가 요구된다.

• Journal of the Korean Wood Science and Technology
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• v.32 no.1
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• pp.59-66
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• 2004

The dried neeldes(1.5 kg) of Juniperus rigida were ground, extracted with acetone-H₂O(7:3, v/v), concentrated, and fractionated with a series of hexane, CH₂Cl₂, EtOAc and water on a separatory funnel. Each fraction was freeze dried to give dark-brown powder and EtOAc and water soluble fractions were chromatographed on a Sephadex LH-20 column using a series of aqueous methanol and ethanol-hexane mixture as eluent. Spectrometric analysis such as NMR and FAB or EI-MS including TLC were performed to characterize the structures of the isolated compounds. The needles of Juniperus rigida contained a large amount of (+)-catechin, quercetin-3-O-α-L-rhamnopyranoside and isoconiferin, in addition to a small amount of umbelliferone and quercetin-3-O-β-D-rutinoside. The antioxidative activities of each fraction and isolated compounds were tested by DPPH radical scavenging method, and EtOAc soluble fraction, (+)-catechin and quercetin-3-O-α-L-rhamnopyranoside were effective.

• Development and Reproduction
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• v.22 no.2
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• pp.175-182
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• 2018

In order to examine the effects of four different light spectra (white, red, green, and blue) on the oocyte maturation, the change of reproductive parameters, via brain-pituitary-gonad (BPG) axis in grass puffer, were investigated. After exposure four different light spectra for 7 weeks, the abundance of gonadotropin-releasing hormone (GnRH) mRNA which is a type of seabream (sbGnRH) and two different subunit of gonadotropin hormones mRNAs, follicle-stimulating hormone ($fsh{\beta}$) mRNA and luteinizing hormone ($lh{\beta}$) mRNA, were analyzed in the brain and pituitary. Histological analysis showed that the mature oocyte ratio in the green spectrum was higher than other light spectra-exposed groups. Gonadosomatic index (GSI) and oocyte developmental stage were also investigated in the gonad based on histological observations. GSI value with the presence of yolk stage oocytes was significantly increased in the green spectrum-exposed group when compared to that of the other light-exposed groups (white, red, and blue) (p<0.05). The abundances of sbGnRH mRNA and $fsh{\beta}$ mRNA in the green spectrum-exposed group were also significant higher than those of the other light spectra-exposed groups (p<0.05). These results indicate that the maturation of oocyte in grass puffer can be accelerated by exposure to the spectrum of green. To better understand the molecular mechanism for the maturation of oocyte in grass puffer, further study examining the relationship between oocyte development and its related genes is required.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1359-1366
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• 2012

A strain of Streptomyces cavourensis subsp. cavourensis (coded as SY224) antagonistic to Colletotrichum gloeosporioides infecting pepper plants was isolated. SY224 produced lytic enzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase, and protease in respective assays. To examine for antifungal activity, the treatments amended with the nonsterilized supernatant resulted in the highest growth inhibition rate of about 92.9% and 87.4% at concentrations of 30% and 10%, respectively. However, the sterilized treatments (autoclaved or chloroform treated) gave a lowered but significant inhibitory effect of about 63.4% and 62.6% for the 10% supernatant concentration, and 75.2% and 74.8% for the of 30% supernatant concentration in the PDA agar medium, respectively, indicative of the role of a non-protein, heat stable compound on the overall effect. This antifungal compound, which inhibited spore germination and altered hyphal morphology, was extracted by EtOAc and purified by ODS, silica gel, Sephadex LH-20 column, and HPLC, where an active fraction was confirmed to be 2-furancarboxaldehyde by GS-CI MS techniques. These results suggested that SY224 had a high potential in the biocontrol of anthracnose in pepper, mainly due to a combined effect of lytic enzymes and a non-protein, heat-stable antifungal compound, 2-furancarboxaldehyde.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.490-497
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• 2010

A chitinolytic bacterium having a strong antagonistic activity against various pathogens including Phytophtora capsici was isolated from rhizosphere soil, and identified as Lysobacter enzymogenes (named as LE429) based on 16S rRNA gene sequence analysis. This strain produced a number of substances such as chitinase, ${\beta}-1$, 3-glucanase, lipase, protease, gelatinase and an antibiotic compound. This antibiotic compound was purified by diaion HP-20, sephadex LH-20 column chromatography and HPLC. The purified compound was identified as phenylacetic acid by gas chromatography-electron ionization (GC-EI) and gas chromatography-chemical ionization (GC-CI) mass spectrometry. In field experiment, pepper plants were treated by the strain LE429 culture (CB), neem oil solution (NO), combination (CB+NO) or control (CON). Plant height and number of branches, flowers and pods of pepper plant in CB treatment were generally highest, and followed by CB+NO, CON and NO. The fungal pathogens were strongly inhibited, while several insect pests were discovered in CB treatment. Any insect pests were not found, while all fungal pathogens tested were not suppressed in NO treatment. However, in CB+NO treatment, non incidence of fungal pathogens and insect pests were found. The strain LE429 producing secondary metabolites with neem oil should be a potential agent to control fungal diseases and insect pests.

• Journal of Korean Society of Forest Science
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• v.95 no.4
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• pp.429-434
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• 2006

The dried needles (1.5 kg) of Picea abies Karsten were ground, extracted with acetone-$H_2O$ (7:3, v/v), concentrated, and fractionated with a series of n-hexane, methylene chloride, ethyl acetate and water on a separatory funnel. Each fraction was freeze dried, then a portion of ethyl acetate soluble powder was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol and ethanol-n-hexane mixture as eluents. The isolated compounds were identified by cellulose TLC, $^1H$-, $^{13}C$-NMR, FAB and EI-MS. (+)-catechin (compound I), (-)-epicatechin (compound II), kaempferol-3-O-${\beta}$-D-glucopyranoside (compound III), 4-hydroxyacetophenone (compound IV) were isolated from the ethyl acetate soluble fraction and (+)-catechin (compound I), protocatechuic acid (compound V) were isolated from the $H_2O$ soluble fraction of P. abies needle. The antioxidative activities of each fraction and the isolated compounds were tested by DPPH radical scavenging method, and EtOAc soluble fraction, (+)-catechin and (-)-epicatechin showed similar values to ${\alpha}$-tocopherol and BHT as controls.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1199-1203
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• 2003

Polyphenols were isolated from Korean green tea using Sephadex LH-20 and HPLC. The isolated polyphenols were procyanidin B-4, procyanidin B-2-3,3'-digallate, prodelphinidin C-2-3,3'-di-O-digallate, (+)-catechin-3-O-rhamnose, procyandin B-5, procyanidin B-7-3-0-gallate, gallate, epiafzelechin-$(4{\beta}{\rightarrow}8)$-epiafzelechin, procyanidin B-3-3-O-rhamnose, afzelechin-$(4{\alpha}{\rightarrow}8)$-catechin, prodelphinidin B-5-3,3'-di-O-digallate and (+)-taxifolin-3-O-D-xyloside. The inhibitory effects of prodelphinidin C-2-3,3'-di-O-gallate and procyanidin B-2-3,3'-digallate $(at\;100{\mu}M)$ on angiotensin.converting enzyme were 68.8 and 54.6%, respectively, while the inhibitory effects of prodelphinidin C-2-3,3'-di-O-gallated and procyanidin B-2-3,3'-digallate $(at\;100{\mu}m)$ on xanthine oxidase were 54.5 and 38.2%, respectively. Lastly, the inhibitory activities of prodelphinidin C-2-3,3'-di-O-gallate $(at\;100{\mu}m)$ on tyrosinase was 42.1%.

• Journal of the Korean Wood Science and Technology
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• v.41 no.6
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• pp.566-575
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• 2013

The fruits of Taxus cuspidata were collected, divided into seeds and fruits, and extracted with 95% EtOH. The extracts were evaporated under the reduced vacuum pressure, concentrated, then successively fractionated with a series of n-hexane, dichloromethane, ethyl acetate and water on a separatory funnel to get some freeze dried samples. A portion of the EtOAc (arils:1.65 g, seeds:1.04 g) and $H_2O$ (arils:7 g, seeds:10 g) soluble samples were chromatographed on a Sephadex column using MeOH-$H_2O$ (1:1, 1:3, 1:5, v/v), EtOH-hexane (3:1, v/v) mixture and 100% $H_2O$ as eluting solvents to isolate pure compounds from the fractions. The isolates were developed by cellulose TLC using t-BuOH-HOAc-$H_2O$ (TBA; 3:1:1, v/v/v) and 6% aqueous HOAc. Visualization was done under ultraviolet light and by spraying the vanillin-HCl-EtOH reagent (4.8:12:480, v/v/v). followed by heating. The structures of the isolates were characterized by $^1H$- and $^{13}C$-NMR, DEPT, 2D-NMR, LC/MS and EI-MS spectra. In addition to the NMR and MS spectra, acid hydrolysis and permethylation were used to determine the correct structure of the isolated sugar compound. Their structures were elucidated as (+)-catechin (1), (-)-epicatechin (2), (+)-gallocatechin (3), (-)-epigallocatechin (4) and ${\beta}$-D-fructofuranose-($2{\rightarrow}4$)-O-${\beta}$-D-glucopyranose($1{\rightarrow}4$)-O-${\alpha}$-D-glucopyranose ($1{\rightarrow}2$)-O-${\beta}$-D-fructofuranose (5) on the basis of the above experimental evidences.