• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.65-70
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• 2006

This study investigated apoptosis and in vitro development of parthenogenetic preimplantation porcine embryos. In vitro matured oocytes for $42{\sim}44h$ were used. Apoptotic cell death was analyzed by using a terminal deoxynucleatidyl transferase mediated deoxyuridine 5-triphosphate nick-end tabling (TUNEL) assay. In experiment 1, oocytes were activated with two electric pulses (CH) of 1.2 kV/cm for $30{\mu}sec$ (E), E + 6-dimethylaminopurine (6-DMAP) or E + cycloheximide (CH) and cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$. In experiment 2, oocytes were activated by E and cultured in PZM-3 or NCSU-23 under a gas atmosphere of 20% $O_2$ ($5%\;CO_2$, in air) or 5% $O_2$ $(5%\;CO_2,\;5%\;O_2\;90%\;N_2)\;at\;38.5^{\circ}C$. Oocytes activated with E+6-DMAP or E+CH showed higher blastocyst rates (36.3% and 32.5%) compared to E alone (27.7%). The frequency of apoptosis according to treatments were 5.3%, 7.7% and 7.1% respectively. Oocytes activated with E alone showed lower (P<0.05) frequency of apoptosis compared to other groups. In experiment 2, parthenotes cultured in PZM-3 showed slightly higher blastocyte rates (28.2% and 29.7%) compared to NCSU-23 (22.6% and 24.4%) regardless of atmosphere. Blastocysts generated in PZM-3 showed lower (P<0.05) apoptosis rate under 20% $O_2$ (9.2% vs 16.9%), whereas those in NCSU-23 had slightly lower apoptosis rate under 5% $O_2$ (14.0% vs 18.4%). This result represents that activation method and culture condition could affect the frequency of apoptosis as well as in vitro developmental rate.

• Journal of the Korean GEO-environmental Society
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• v.9 no.6
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• pp.13-19
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• 2008

The objectives of this paper are to investigate states of these open dumping landfills for managing, utilizing and stabilization of a municipal solid waste landfill site in environmentally secure conditions. The result of the physical analysis of the landfill showed that it is composed of between 29.72-63.84% organic matter, 32.88% vinyl plastic. The environmental assessment of the landfill site confirmed that the landfill is at a maturation phase due to 0.18 of $BOD_5$/CODcr of leachate. VS and FS of TS Was respectively 30.37%, 32.34% and C/N ratio was 21.8. Surface water around landfill was BOD 10.7 mg/g, SS 37.8 mg/g, E-Coli 31,157(MPN/$100m{\ell}$) and Ground water was $COD_{Mn}$ 1.13-1.38 mg/g, $NO_3-N$ 1.025-4.075 mg/g. Leachat indicated T-P 0.002-0.028 mg/g, $NH_3-N$ 4.0-21.0 mg/g. The soil contamination of around landfill didn't appear as below of the regulation of Soil Environment Conservation Act. The Landfill Gas was $CH_4$ 13.25%, $CO_2$ 6.17%, H2S and CO was not detected. Also Surface Water was not detected $CO_2$, $CH_4$, $H_2S$, CO.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.1010-1016
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• 2000

Effect on the bio-mass and nutrient yield of Lemna perpusilla Torry was studied at two nutrient loading frequencies. Six polythene sinks ($6{\times}2{\times}0.5m^{3}$ area) randomly allocated to two treatments having three sinks in each treatment. Each sink containing $6{\times}2{\times}0.35m^{3}$ water body, was changed either daily with 12.6 kg anaerobically fermented cattle manure (effluent) or with 76 kg efluent at six-day interval. Media Kjeldhal-N (40 vs. 36 mg/L; p>0.05) and $NH_3$-N (9 vs 7 mg/L; p<0.05) concentration were higher in daily charged sinks than those charged at six-day interval. However, temperature ($28.48{^{\circ}C}\;vs.\;28.60{^{\circ}C}$) and pH (6.82 vs. 6.84) were sinilar in both groups of sinks. Biomass (93 vs. $80g/m^{2}/d$) and crude protein (1.24 vs. $1.11g/m^{2}/d$) yield were non-significantly (p>0.05) higher in the daily charged than 6-d interval charged ponds. However, dry matter (4.52 vs. 4.15%), organic matter (83 vs. 84%), crude protein (31.4 va. 29.6%) and organic carbon (47 vs. 47%) content were similar in both daily and 6-d interval charged ponds. There were no apparent relationships between the crude protein content of Lemna and the media Kjeldhal-N or $NH_3$-N concentration apparently due to saturated level of nutrient (N in particular). It was concluded that with the level of effluent used for two treatments, nutrient-loading frequency had no significant effect on the biomass and protein yield of Lemna. Technique described here can be used for year-round duckweed production from fermented cattle manure for feeding poultry/fish in smallholder production system.

• Journal of Plant Biotechnology
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• v.40 no.3
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• pp.169-177
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• 2013

Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus ${\rightarrow}$ tobacco E2 signal peptide ${\rightarrow}$ B-peptide (1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-peptide (1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.47-55
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• 2011

The solvent extracts of Aruncus dioicus var. kamtschaticus Hara, which were extracted by using several solvents with different polarities, were performed to investigate the antioxidant activities, whitening effect and antimicrobial activity. The content of total polyphenol of fractions from Aruncus dioicus var. kamtschaticus Hara extract showed the highest value ($335.88{\pm}2.26$ mg/g GAE) on ethyl acetate fraction. The ethyl acetate and n-butanol fractions were 0.06 mg/mL and 0.25 mg/mL as $IC_{50}$ values on DPPH radical scavenging, and $99.16{\pm}0.09%$ and $89.29{\pm}0.64%$ on ABTS radical scavenging activity, respectively. Also, reducing power and FRAP value were significantly higher on ethyl acetate fraction. The SOD like activity showed $80.76{\pm}0.61%$ on ethyl acetate and $72.34{\pm}0.79%$ on n-butanol. Tyrosinase inhibition activities (at 5 mg/mL) were $59.08{\pm}0.98%$ on ethyl acetate fraction. The chloroform fraction showed the strongest antimicrobial activities against B. cereus (14 mm), B. subtilis (12.5 mm), S. aureus (10.8 mm), E. coli (20.7 mm) at 0.1 mg/disc and the inhibition zone diameter of ethyl acetate fraction was 17.2 mm against E. coli at 0.5 mg/disc. The minimum inhibitory concentrations (MIC) of chloroform fraction against B. cereus and E. coli were 50 and $25{\mu}g$/mL, respectively. From these results, it is suggested that ethyl acetate and n-butanol fractions of Aruncus dioicus var. kamtschaticus Hara could be used as functional material for food additive ingredient and chloroform fraction could be suitable for the development of a food preservative.

• Korean Journal of Acupuncture
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• v.29 no.2
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• Journal of the Korean Chemical Society
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• v.29 no.6
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• pp.629-636
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• 1985

The rates of solvolysis of trans-$[Co(AA)_2Cl_2]^+$ in which AA indicates ethylenediamine(en), N-ethylethylenediamine (N-eten), N-methylethylenediamino (N-meen) and trimethylenediamine(tn) respectively have been investigated using conductometric and spectrophotometric methods at various pressure up to 2,000 bar in acetone-water mixture. The activation volumes (${\Delta}V^{\neq}) obtained from the pressure effect on rate constants were -0.2∼0.9 $cm^3mole^{-1}$ for en, -0.2∼0.6 $cm^3mole^{-1}$ for N-eten, -0.8∼6.0 $cm^3mole^{-1}$ for N-meen and 0.7∼7.0$cm^3mole^{-1}$ for tn. The rates of solvolysis of these complexes were analyzed by comparing with the results obtained from excess free energy ($G^E$) and free energy cycle. It was found that $S_N1$ character was increased with decreasing the pressure and increasing the content of acetone in the mixture solvent. In addition to that, the effect of charge separation on the mechanism of solvolysis was discussed.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.1
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• pp.29-35
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• 2005

To prove the buffering contribution of mitochondria to the increase of intracellular $Ca^{2+}$ level ($[Ca^{2+}]_i$) via background nonselective cation channel (background NSCC), we examined whether inhibition of mitochondria by protonophore carbonylcyanide m-chlorophenylhydrazone (CCCP) affects endothelial $Ca^{2+}$ entry and $Ca^{2+}$ buffering in freshly isolated rabbit aortic endothelial cells (RAECs). The ratio of fluorescence by fura-2 AM ($R_{340/380}$) was measured in RAECs. Biological state was checked by application of acetylcholine (ACh) and ACh ($10{\mu}M$) increased $R_{340/380}$ by $1.1{\pm}0.15$ ($mean{\pm}S.E.$, n=6). When the external $Na^+$ was totally replaced by $NMDG^+$, $R_{340/380}$ was increased by $1.19{\pm}0.17$ in a reversible manner (n=27). $NMDG^+$-induced $[Ca^{2+}]_i$ increase was followed by oscillatory decay after $[Ca^{2+}]_i$ reached the peak level. The increase of $[Ca^{2+}]_i$ by $NMDG^+$ was completely suppressed by replacement with $Cs^+$. When $1{\mu}M$ CCCP was applied to bath solution, the ratio of $[Ca^{2+}]_i$ was increased by $0.4{\pm}0.06$ (n=31). When $1{\mu}M$ CCCP was used for pretreatment before application of $NMDG^+$, oscillatory decay of $[Ca^{2+}]_i$ by $NMDG^+$ was significantly inhibited compared to the control (p＜0.05). In addition, $NMDG^+-induced$ increase of $[Ca^{2+}]_i$ was highly enhanced by pretreatment with $2{\mu}M$ CCCP by $320{\pm}93.7$%, compared to the control ($mean{\pm}S.E.$, n=12). From these results, it is concluded that mitochondria might have buffering contribution to the $[Ca^{2+}]_i$ increase through regulation of the background NSCC in RAECs.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.2
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• pp.599-606
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• 2012

The characterization of the whole genome of human papillomavirus type 16 (HPV16) from cervical cancer specimens with multiple infections in comparison with single infection samples as the oncogenic potential of the virus may differ. Cervical carcinoma specimens positive for HPV16 by PCR and INNO-LiPA were randomly selected for whole genome characterization. Two HPV16 single infection and six HPV16 multiple infection specimens were subjected to whole genome analysis by using conserved primers and subsequent sequencing. All HPV16 whole genomes from single infection samples clustered in the European (E) lineage while all multiple infection specimens belonged to the non-European lineage. The variations in nucleotide sequences in E6, E7, E2, L1 and Long control region (LCR) were evaluated. In the E6 region, amino acid changes at L83V were related to increased cancer progression. An amino acid variation N29S within the E7 oncoprotein significantly associated with severity of lesion was also discovered. In all three domains of the E2 gene non synonymous mutations were found. The L1 region showed various mutations which may be related to conformation changes of viral epitopes. Some transcription factor binding sites in the LCR region correlated to virulence were shown on GRE/1, TEF-1, YY14 and Oct-1. HPV16 European variant prone to single infection may harbor a major variation at L83V which significantly increases the risk for developing cervical carcinoma. HPV16 non-European variants prone to multiple infections may require many polymorphisms to enhance the risk of cervical cancer development.

• Electrical & Electronic Materials
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• v.8 no.2
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• pp.171-175
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• 1995

Optimum condition of remote plasma enhanced chemical vapor deposition using orthogonal array method was chosen. Characteristics of oxide films deposited by RPECVD with SiH$_{4}$ and N$_{2}$O gases were investigated. Etching rate of the optimized SiO$_{2}$ films in P-etchant was about 6[A/s] that was almost the same as that the high temperature thermal oxide. The films showed high dielectric breakdown field of more than 7[MV/cm] and a resistivity of 8*10$^{13}$ [.ohmcm] around at 7[MV/cm]. The interface trap density of SiO$_{2}$/Si interface around the midgap derived from the high frequency C-V curve was about 5*10$^{10}$ [/cm$^{2}$eV]. It was observed that the dielectric constant of the optimized SiO$_{2}$ film was 4.29.