• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.31-31
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• 2001

This study was to establish in uitro culture system of mouse preantral follicles and to obtain higher in vitro development rates and production of live young. Preantral follicles were obtained from 12-day-old FI mouse (C57BL $\times$ CBA) by enzymatical methods. Oocyte-granulosa cell complexes (OGCs) of preantral follicles were loaded on Transwell-COL insert and cultured in $\alpha$MEM supplemented with 5% FBS, 100 mIU/$m\ell$ FSH and 100 mIU/$m\ell$ hMG for IVG. IVM was performed in $\alpha$MEM supplemented 1.5 IU/$m\ell$ hCG for 18 hrs and IVF was carried out in Ml6 medium. Embryos were cultured in modified Ml6 medium supplemented 10% FBS for 4 days. The effect of the OGCs size on the nuclear/cytoplasmic maturation was significantly higher in 120-150 ${\mu}{\textrm}{m}$ (MII: 33.0%, $\geq$2-cell: 36.7%, $\geq$morula: 20.9%) than in 70-110 ${\mu}{\textrm}{m}$ (MII: 12.2%, $\geq$2-cell: 10.2%, $\geq$morula: 4.8%) (p<0.001). In period of the IVG days, the rate of $\geq$2-cell was significantly higher in 10 days(38.2%) than in 12 days (20.0%) (p<0.01). In period of IVF time, 9 hrs ($\geq$2-cell: 31.5%, $\geq$ morula: 14.3%) indicated significantly higher cytoplasmic maturation rate than 4 hrs ($\geq$2-cell: 17.5%, This study was to establish in vitro culture system of mouse preantral follicles and to obtain higher in vitro development rates and production of live young. Preantral follicles were obtained from 12-day-old FI mouse (C57BL $\times$ CBA) by enzymatical methods. Oocyte-granulosa cell complexes (OGCs) of preantral follicles were loaded on Transwell-COL insert and cultured in $\alpha$MEM supplemented with 5% FBS, 100 mIU/$m\ell$ FSH and 100 mIU/$m\ell$ hMG for IVG. IVM was performed in $\alpha$MEM supplemented 1.5 IU/$m\ell$ hCG for 18 hrs and IVF was carried out in Ml6 medium. Embryos were cultured in modified Ml6 medium supplemented 10% FBS for 4 days. The effect of the OGCs size on the nuclear/cytoplasmic maturation was significantly higher in 120-150 ${\mu}{\textrm}{m}$ (MII: 33.0%, $\geq$2-cell: 36.7%, $\geq$morula: 20.9%) than in 70-110 ${\mu}{\textrm}{m}$ (MII: 12.2%, $\geq$2-cell: 10.2%, $\geq$morula: 4.8%) (p<0.001). In period of the IVG days, the rate of $\geq$2-cell was significantly higher in 10 days(38.2%) than in 12 days (20.0%) (p<0.01). In period of IVF time, 9 hrs ($\geq$2-cell: 31.5%, $\geq$ morula: 14.3%) indicated significantly higher cytoplasmic maturation rate than 4 hrs ($\geq$2-cell: 17.5%, $\geq$morula: 4.8%) and 7 hrs ($\geq$2-cell: 20.4%, $\geq$morula: 6.1%) (p<0.01). However, there was no difference in cytoplasmic maturation between co-cultured preantral follicle ( $\geq$morula: 17.4%) and preantral follicle cultured in Ml6 ( $\geq$morula: 17.4%). 22 morula and blastocysts produced in above optimal condition were transferred to uterus of 2 pseudopregnant recipients, 1 recipient was pregnant and then born 1 live young. This result demonstrates that in vitro culture system of preantral follicles can be used efficiently as another method to supply mouse oocyte.morula: 4.8%) and 7 hrs (2-cell: 20.4%, $\geq$morula: 6.1%) (p<0.01). However, there was no difference in cytoplasmic maturation between co-cultured preantral follicle ( $\geq$morula: 17.4%) and preantral follicle cultured in Ml6 ( $\geq$morula: 17.4%). 22 morula and blastocysts produced in above optimal condition were transferred to uterus of 2 pseudopregnant recipients, 1 recipient was pregnant and then born 1 live young. This result demonstrates that in vitro culture system of preantral follicles can be used efficiently as another method to supply mouse oocyte.

• IMMUNE NETWORK
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• v.20 no.5
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• pp.40.1-40.15
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• 2020

The protein encoded by the Gene Associated with Retinoid-Interferon-Induced Mortality-19 (GRIM-19) is located in the mitochondrial inner membrane and is homologous to the NADH dehydrogenase 1-alpha subcomplex subunit 13 of the electron transport chain. Multiple sclerosis (MS) is a demyelinating disease that damages the brain and spinal cord. Although both the cause and mechanism of MS progression remain unclear, it is accepted that an immune disorder is involved. We explored whether GRIM-19 ameliorated MS by increasing the levels of inflammatory cytokines and immune cells; we used a mouse model of experimental autoimmune encephalomyelitis (EAE) to this end. Six-to-eight-week-old male C57BL/6, IFNγ-knockout (KO), and GRIM-19 transgenic mice were used; EAE was induced in all strains. A GRIM-19 overexpression vector (GRIM19 OVN) was electrophoretically injected intravenously. The levels of Th1 and Th17 cells were measured via flow cytometry, immunofluorescence, and immunohistochemical analysis. IL-17A and IFNγ expression levels were assessed via ELISA and quantitative PCR. IL-17A expression decreased and IFNγ expression increased in EAE mice that received injections of the GRIM-19 OVN. GRIM19 transgenic mice expressed more IFNγ than did wild-type mice; this inhibited EAE development. However, the effect of GRIM-19 overexpression on the EAE of IFNγ-KO mice did not differ from that of the empty vector. GRIM-19 expression was therapeutic for EAE mice, elevating the IFNγ level. GRIM-19 regulated the Th17/Treg cell balance.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.2
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• pp.187-198
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• 1997

This experiment was conducted to evaluate effects of wilting and additives on fermetation characteristics, quality and aerobic stability of silage of rye(Seca1e cereale L.) harvested at the early heading stage. Harvested rye was wilted or mixed with beet pulp and then treated with formic acid or lactic acid bacteria(LAB) inoculant. Treatments were consisted of direct cut(DC), wilted(WT), beet pulp added(BP), direct cut and formate applied(DF), wilted and formate applied(WF), beet pulp and formate applied(BF), direct cut and LAB inoculated(DL), beet pulp added and LAB inoculated(BL), and wilted and LAB inoculated(WL) silages. Afler 70 days of ensiling period, the silages were opened and exposed to air for 7 days, and the silages of opening day(0 day) were compared with the silages exposed to air for 7 days(7 day). 1. LAB inoculated rye silages(DL, WL, BL) lowered pH values effectively eom the third day of ensiling and showed most stable pattern of pH changes during the initial fermentation process. Direct cut and formate applied(DF), beet pulp and formate applied(BF) and beet pulp added(BP) silages were also effective in lowering pH from the seventh day thai direct cut treatment(DC). 2. pH was low below 3.7 in all treatments of 0 day. After 7-day period of aerobic exposure, DL and WL showed considerable increases in pH from 3.5 and 3.4 to 8.3 and 6.4, respectively. 3. Direct cut rye silages(DF, DC, DL) and beet pulp and formate treatment(BF) produced effluent of 121.2, 85.9, 80.3 and 34.2 mlkg, respectively and these were greater than others(P<0.01). Beet pulp retained 1.61 I/kg of effluent at the application rate of 50 kg/t in comparing BP with DC, and formate application increased the amount of effluent in comparing DF and BF with DC and BP, respectively(P<0.01). 4. Beet pulp and LAB treatment(B1) was highest in content of lactate as 3.1% and formate treated silages(DF, WF, BF) showed lower content of lactate and total acid than others. LAB inoculated(D1, WL, BL) and wilted (WT) silages were graded to be good quality as a second group by the Flieg's score. Formate applied silages (DF, WF, BF) were graded as a foum group lower than DC by one. 5. Residual content of WSC of rye silage was higher in BP, BF, DL and BL than direct cut treatment(M3) (P< 0.01). Formate application had a tendency to increase the content of residual content of WSC. 6. For the ratio of NH3-N to Total N, wilted and LAB inoculated(W1) and direct cut and LAB inoculated(D1) silages were lowest on 0 day as 8.9 and 9.3% respectively. But after 7-day period of aerobic exposure, WL and DL showed largest increase of the ratio of $NH_3-N$ to Total N from 0 day(P<0.01). 7. On 0 day wilted(WT), beet pulp and formate treatment(BF), and beet pulp and LAB treatment(B1) were lower than direct cut treatment(DC) for NDF and ADF concentrations consistently. Formate applied silages (DF, WF, BF) made little change in ADF and NDF concentrations during 7 day period of aerobic exposure. DL and WL showed a large increase in ADF and NDF concentrations under aerobic condition(P<0.01). 8. IVDMD values of wilted(WT), beet pulp added(BP), wilted and LAB inoculated(W1) and beet pulp added and LAB inoculated(B1) silages were higher than direct-cut treatmentPC) as 84.7, 84.7, 84.4 and 83.0%, respectively on 0 day. But during 7-day period of aerobic exposure, a great decreae in IVDMD of WL was showed(P<0.01). The experimental results indicate that wilted silage(WT) could be recommended as the most effective treatment for reducing efluent and increasing quality and feed value of rye silage without deteriorating aerobic stability more than direct cut treatment(DC). Additionally, under unfavorable weather condition beet pulp added and LAB inoculated treatment(T3L) might be the possible alternative for successful ensiling of forage rye.

• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1965-1969
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• 2007

Triamino ${\alpha}$-cyclodextrin (CD) was synthesized and the inclusion complex with Minoxidil (MXD) was prepared. ${\alpha}$-CD was azidated by modifying the 6-hydroxylmethyl CD rim with sodium azide. Then, mono-, di-, tri-, and tetra-azidocyclodextrins were separated by a flash column chromatography and reduced to the corresponding amines by hydrogenation with Pd/C. The substantivities of MXD included in either 2-hydroxypropyl ${\alpha}$-CD (HP ${\alpha}$-CD) or triamino ${\alpha}$-CD were evaluated in vitro using hairless mice skins. After applying the preparations onto the skin and rinsing it, the amount of the drug left on the skin was determined using high-performance liquid chromatography (HPLC). It was the highest when the drug was included in triamino ${\alpha}$-CD. The electrostatic interaction between the protonated amino CD and the negatively charged skin would be responsible for the relatively high substantivity. The in vivo hair growth promotion effect of each preparation was investigated, where the sample application onto the clipped backs of female mice (C57BL6) and the subsequent rinsing of the backs were done once a day for 30 days. Only MXD in triamino ${\alpha}$-CD had hair growth promotion effect, possibly due to the significant substantivity.

• Genomics & Informatics
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• v.4 no.1
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• pp.23-32
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• 2006

In order to investigate the molecular basis of the aging process in brain, we have employed high-density oligonucleotide microarrays providing data on 10,108 gene clusters to define transcriptional patterns in three brain regions, cerebral cortex, cerebellum, and hippocampus. Comparison of the expression patterns between young (6-week-old) and aged (17-month-old) C57BL/6 male micerevealed that about ten percent (1098) of the genes showed a significant change in the expression level in at least one of the three tissues. Among them, 23 genes were upregulated and 62 genes were downregulated in all three tissues of the old mice. The number of genes upregulated exclusively in hippocampus (337) was much larger compared to other tissues. Gene ontology-based analysis showed the genes related with signal transduction or molecular transports are more likely to be upregulated than downregulated in the aging process of hippocampus. These data may provide some useful means for elucidating the molecular aspect of aging in hippocampus and other regions in brain.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.225-229
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• 2005

To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL 17) isolated from B. flavum transformed recipient $McrBC^+$ strains of Escherichia coli with lower efficiency than $McrBC^-$. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBc sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient $EcoKR^+$ strains of E. coli with lower efficiency than $EcoKR^-$, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.

• The Korean Journal of Food And Nutrition
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• v.21 no.1
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• pp.7-14
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• 2008

The study was performed to investigate the biological activity of Enteromorpha intestinalis. In order to examine its blood anti-coagulant effects, Enteromorpha intestinalis was extracted with cold water, methanol, hot water, HCl and NaOH. In general, the alkali extract of Enteromorpha intestinalis was approximately 17 times stronger than the control. The anti-cancer effects of select extracts(methanol, hot water, 0.1 N NaOH, 1 N NaOH) were determined in human melanoma cells(Bl6/F10), fibrosarcoma cells(HTl080) and breast cancer cells(MCF7) by MTT assay. With the treatment of 250 ${\mu}g/m{\ell}$ of methanol extracts. HT1080, B16/F10 and MCF7 cell viabilities significantly decreased to 8.06%, 3.62% and 10.10%, respectively. Thus these results strongly support the possibie use of Enteromorpha intestinalis as a functional materials.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.116-122
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• 2007

An efficient enzyme system for the synthesis of L-tyrosine was developed using a fed-batch reactor with continuous feeding of phenol, pyruvate, and ammonia. A thermo- and chemostable tyrosine phenol-lyase from Symbiobacterium toebii was employed as the biocatalyst in this work. The enzyme was produced using a constitutive expression system in Escherichia coli BL21, and prepared as a soluble extract by rapid clarification, involving treatment with 40% methanol in the presence of excess ammonium chloride. The stability of the enzyme was maintained for at least 18 h under the synthesis conditions, including 75 mM phenol at pH 8.5 and $40^{\circ}C$. The fed-batch system (working volume, 0.51) containing 1.0 kU of the enzyme preparation was continuously fed with two substrate preparations: one containing 2.2 M phenol and 2.4 M sodium pyruvate, and the other containing 0.4 mM pyridoxal-5-phosphate and 4M ammonium chloride (pH 8.5). The system produced 130g/I of L-tyrosine within 30h, mostly as precipitated particles, upon continuous feeding of the substrates for 22 h. The maximum conversion yield of L-tyrosine was 94% on the basis of the supplied phenol.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1805-1810
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• 2007

Purple nonsulfur bacteria were isolated from river sediments and their growth promoting capabilities on tomato were examined. Isolated strains KL9 and BL6 were identified as Rhodopseudomonas spp. by 16S rDNA sequence analysis. Rhodopseudomonas strain KL9 maximally produced 5.56 mM/min/mg protein and $67.2\;{\mu}M/min/mg$ protein of indole-3-acetic acid (IAA) and 5-aminolevulinic acid (ALA), respectively, which may be one of the mechanisms of plant growth enhancement. The germination percentage of tomato seed, total length, and dry mass of germinated tomato seedling increased by 30.2%, 71.1%, and 270.8%, respectively, compared with those of the uninoculated control 7 days after inoculation of strain KL9. The lengths of the root and shoot of germinated seedling treated with 3 mM tryptophan, a precursor of IAA, increased by 104.4% and 156.5%, respectively, 7 days after inoculation of strain KL9. Rhodopseudomonas KL9 increased 123.5% and 54% of the root and shoot lengths of germinated seedling, respectively, treated with 15 mM glycine and succinate, precursors of ALA. This plant growth promoting capability of purple nonsulfur bacteria may be a candidate for a biofertilizer in agriculture.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.19 no.2
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• pp.202-211
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• 2006

Objectives : This study is designed to evaluate correlation between the data of D.I.T,I examination in Bell's palsy and the improving period. Methods : This study researched into the clinical statistics for 50 patients who are in Bell's palsy and they are treated with oriental medical care of Dongshin university during 18months from January 1, 2005 to June 1, 2006. The data which examinated by D.I.T.I, are the thermal difference of normal and abnormal site of acupuncture int Gl4 BL2 GB3 S3 S4 S6 TE17 in Bell's palsy. Result : There was statistic significance between thermal difference of normal, abnormal site and the improving period. The period was shorter in normal thermal pattern than hypothermal pattern, hyperthermal pattern. Conclusion : According to the result, there was a statistic significance that more thermal difference and more difficult to recover in Bell's palsy.