• Microbiology and Biotechnology Letters
• /
• v.40 no.4
• /
• pp.348-355
• /
• 2012

In the present study, we investigated the overexpression and characterization of bovine pancreatic (bp)- DNase I in Saccharomyces cerevisiae and Pichia pastoris. The bp-DNase I gene was fused in frame with the GAL10 promoter, $MF{\alpha}$, and GAL7 terminator sequences, resulting in the plasmid, pGAL-$MF{\alpha}$-DNaseI (6.4 kb). Also, the bp-DNase I gene was fused in frame with the AOX1 promoter, $MF{\alpha}$, and AOX1 terminator sequences, resulting in the plasmid, pPEXI (8.8 kb). The recombinant plasmids, pGAL-$MF{\alpha}$-DNaseI and pPEXI were introduced into S. cerevisiae and P. pastoris host cells, respectively. When the transformed yeast cells were cultured at $30^{\circ}C$ for 48 h in galactose or methanol medium, bp-DNase I was overexpressed and the most of activity was found in the extracellular fraction. P. pastoris transformant activity showed 45.5 unit/mL in the culture medium at 48 h cultivation, whereas S. cerevisiae transformant revealed 37.7 unit/mL in the extracellular fraction at 48 h cultivation. The enzymatic characteristics, such as DNA cleavage and half life were investigated. Treatment of the recombinant DNase I from P. pastoris induced degradation of the calf thymus DNA within 1 minute, and this DNA degradation rate was higher than that of commercial bp-DNase I (SIGMA) and the recombinant DNase I from S. cerevisiae.

• The Journal of the Petrological Society of Korea
• /
• v.28 no.1
• /
• pp.1-13
• /
• 2019

The characteristics of the rock cleavage of Jurassic Geochang granite were analysed using the parameters from the length and spacing-cumulative frequency diagrams. The evaluation for three planes and three rock cleavages was performed using the 25 parameters such as (1~2) slope angle(${\alpha}^{\circ}$and ${\beta}^{\circ}$), (3) intersection angle(${\alpha}-{\beta}^{\circ}$), (4) exponent difference(${\lambda}_S-{\lambda}_L$), (5~12) length of line(oa, ob, ol, os, ss', ll' and sl') and (13~15) length ratio(ol/os, ss'/ll' and ll'/sl'), (16) mean length((ss'+ll')/2), (17~23) area (${\Delta}oaa^{\prime}$, ${\Delta}obb^{\prime}$, ${\Delta}obb^{\prime}$, ${\Delta}oaa_a^{\prime}$, ${\Delta}obb_a^{\prime}$, ${\Delta}ll^{\prime}s^{\prime}$, ${\Delta}ss^{\prime}l^{\prime}$ and ⏢$ll^{\prime}ss^{\prime}$) and (24~25) area difference(${\Delta}obb^{\prime}-{\Delta}oaa^{\prime}$ and ${\Delta}obb_a^{\prime}-{\Delta}oaa_a^{\prime}$). Firstly, the values of the 11 parameters(group I: No. 1, 3~4, 7, 9~10, 13, 15~16, 20 and 25), the 3 parameters(group II: No. 5, 8 and 17) and the 2 parameters(group III: No. 12 and 22) are in orders of H(hardway) < G(grain) < R(rift), R < G < H and G < H < R, respectively. On the contrary, the values of parameters belonging to the above three groups show reverse orders for three planes. Secondly, the generalized chart for three planes and three rock cleavages were made. From the related chart, the distribution types formed by the two diagrams related to lengths and spacings were derived. The diagrams related to spacings show upward curvature in the chart of rift plane(G1 & H1, R') and hardway(H1 & H2, H). On the contrary, the diagrams related to lengths show downward curvature. These two diagrams take the form of a convex lens in the upper section. Besides, the two diagrams cross each other in the lower section. The overall shape formed by the above two diagrams between three planes($H^{\prime}{\rightarrow}G^{\prime}{\rightarrow}R^{\prime}$) and three rock cleavages($R{\rightarrow}G{\rightarrow}H$) display in reverse order. Lastly, these types of correlation analysis is useful for discriminating three quarrying planes.

• Journal of Microbiology
• /
• v.37 no.2
• /
• pp.73-79
• /
• 1999

Pseudomonas sp. NGK1, isolated by naphthalene enrichment culture technique, is capable of degrading anthracene as a sole source of carbon and energy. The organism degraded anthracene through the intermediate formation of 1,2-dihydroxyanthracene, 2-hydroxy-3-naphthoic acid, salicylate, and catechol. The intermediates were isolated and characterized by TLC, spectrophotometry, and HPLC analysis. The cell free extract of anthracene-grown cells showed activities of anthracene dioxygenase, 2-hydroxy-3-naphthylaldehyde dehydrogenae, 2-hydroxy-3-naphthoate hydroxylase, salicylate hydroxylase and catechol 2,3-dioxygenase. The formed catechol as a metabolite is degraded through meta-cleavage with the formation of ${\alpha}$-hydroxymuconic semi-aldehyde.

• Biomedical Science Letters
• /
• v.20 no.3
• /
• pp.168-172
• /
• 2014

It has recently been shown that hypothermia treatment improves brain ischemia injury and is being increasingly considered by many clinicians. However, the precise roles of hypothermia for brain ischemia are not yet clear. In the present study we demonstrated firstly that hypothermia induced beta-catenin-interacting protein 1 (CTNNBIP1) gene expression and its expression was dramatically decreased under ischemic conditions. It was also demonstrated that hypothermia activated endoplasmic reticulum (ER) stress sensors especially both, the phosphorylation of $eIF2{\alpha}$, and ATF6 proteolytic cleavage. However, the factors of apoptosis and autophagy were not associated with hypothermia. These findings suggested that hypothermia controlled CTNNBIP1 gene expression under ischemia, which may provide a clue to the development of treatments and diagnostic methods for brain ischemia.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.1
• /
• pp.219-224
• /
• 2011

The new MPV type reagent, Al-trifluoromethanesulfonyldiisobutylalane ($DIBAO_3SCF_3$), has been prepared and its reducing characteristics in the reduction of selected organic compounds containing representative functional groups have been examined, and compared its reactivity with that of Al-methanesulfonyldiisobutylalane ($DIBAO_3SCH_3$) in order to understand the fluorine-substituent effect on its reactivity. In general, the reactivity of $DIBAO_3SCF_3$ appears to be much higher than that of $DIBAO_3SCH_3$, apparently due to the acidity increase by the electron-withdrawing fluorine-substituent. The reagent reduced aldehydes and ketones readily, but showed a perfect selectivity in the reduction of $\alpha,\beta$-unsaturated aldehydes and ketones to produce the corresponding allylic alcohols in an absolutely 100% purity. In addition, the reagent achieved the regioselective cleavage of phenyl- or/and alkyl-substituted epoxides to the less substituted alcohols in a perfect regioselectivity. Moreover, the reagent also showed an high stereoselectivity in the reduction of substituted cycloalkanones to produce the thermodynamically more stable alcohol epimers exclusively.

• Journal of the Korean Society of Safety
• /
• v.18 no.3
• /
• pp.46-53
• /
• 2003

For the tensile tests of the F.E.M., microvoids are created by the boundary separation process at the martensite boundary or neighborhood and at inclusions within the fracture. to grow to the ductile dimple fracture. For the case of the M.E.F., microvoids created at the discontinuities of the martensite phase which exists at the grain boundary of the primary ferrite are grown to coalescence with the cleavage cracks induced at the interior of the ferrite, which as a result show the discontinuous brittle fracture behavior. In spite of their similar tensile strengths, the fatigue limit and the notch sensitivity of the M. E.F. is superior to those of the F.E.M., The M.E.F. is much more insensitive to notch than F.E.M. from the stress concentration factor($\alpha$).

• Journal of Embryo Transfer
• /
• v.31 no.3
• /
• pp.199-205
• /
• 2016

The aim of this study was to investigate the effect of uterine histotroph on embryo development and the expression of cysteine-rich protein 2 (CRP2), coatomer subunit gamma-2 (G2COP), myoglobin (MYG), vascular endothelial growth factor D (VEGFD), collagen alpha 4 chain (COL4) and galactoside 3-L-fucosyltransferase 4 (FUT4) proteins in porcine embryo during pre-implantation. Uterine histotroph (UH) was collected from uterine horn on corpus albican phase, and embryos were cultured in porcine zygote medium with UH for 168 hours. Cleavage and blastocyst formation of embryo were detected at 168 hours after in vitro fertilization. And CRP2, G2COP, MYG, VEGFD, COL4 and FUT4 proteins were observed using confocal laser microscope. In results, embryo cleavage rate was not significantly changed by UH, but blastocyst rate was significantly (P<0.05) decreased in UH-treated embryos. Moreover, CRP2, G2COP, MYG, VEGFD, COL4 and FUT4 proteins were expressed in blastomere. CRP2 in embryo was significantly overexpressed (P<0.05), but not G2COP, MYG, VEGFD, COL4 and FUT4 proteins. In summary, UH on corpus albican phase was increased CRP2 protein in embryo, and inhibited blastocyst formation in preimplantation porcine embryos, suggesting that CRP2 may play an interrupter on embryo development in pigs.

• Bulletin of the Korean Chemical Society
• /
• v.23 no.9
• /
• pp.1229-1258
• /
• 2002

This paper reports the photochemical reduction of benzil 1 to benzoin 2 and the reduction of 2 to hydrobenzoin 4 in deoxygenated solvents in the presence of triethylamine (TEA) and/or TiO2. Without TEA or TiO2, the photolysis of 1 resulted in very low yield of 2. The presence of TEA or TiO2 increased the rate of disappearance of 1 and the yield of 2, which were further increased considerably by the presence of water. The photoreduction of 1 to 2 proceeds through an electron transfer to 1 from TEA or hole-scavenged excited TiO2 followed by protonation. In the reaction medium of 88 : 7 : 2 : 3 CH3CN/CH3OH/H2O/TEA with 2.5 $㎎/m{\ell}$ of TiO2, the yield of 2 was as high as 85 % at 50 % conversion of 1. The photolysis of 2 in homogeneous media resulted in photo-cleavage to benzoyl and hydroxybenzyl radicals, which are mostly converted to benzaldehyde. The reduction product 4 is formed in low yield through the dimerization of hydroxybenzyl radicals. The addition of TEA increased the conversion rate of 2 and the yield of 4 significantly. This was attributed to the scavenging effect of TEA for benzoyl radical to produce N,N-diethylbenzamide and the photoreduction of benzaldehyde in the presence of TEA. The ratio of $(\pm)$ and meso isomers of 4 obtained from the photochemical reaction is about 1.1. This ratio is the same as that from the photochemical reduction of benzaldehyde in the presence of TEA. In the TiO2-sensitized photochemical reduction of 2, meso-4 was obtained in moderate yield. The reduction of 2 to 4 proceeds through two consecutive electron/proton transfer processes on the surface of the photocatalyst without involvement of ${\alpha}-cleavage$. The radical 11 initially formed from 2 by one electron/proton process can also combine with hydroxy methyl radical, which is generated after hole trapping of excited TiO2 by methanol, to produce 1,2-diphenylpropenone after dehydration reaction.

• The Korean Journal of Physiology and Pharmacology
• /
• v.22 no.3
• /
• pp.349-360
• /
• 2018

Autophagy has been studied as a therapeutic strategy for cardiovascular diseases. However, insufficient studies have been reported concerning the influence of vascular smooth muscle cells (VSMCs) through autophagy regulation. The aim of the present study was to determine the effects of VSMCs on the regulation of autophagy under in vitro conditions similar to vascular status of the equipped micro-tubule target agent-eluting stent and increased release of platelet-derived growth factor-BB (PDGF-BB). Cell viability and proliferation were measured using MTT and cell counting assays. Immunofluorescence using an $anti-{\alpha}-tubulin$ antibody was performed to determine microtubule dynamic formation. Cell apoptosis was measured by cleavage of caspase-3 using western blot analysis, and by nuclear fragmentation using a fluorescence assay. Autophagy activity was assessed by microtubule-associated protein light chain 3-II (LC-II) using western blot analysis. Levels of intracellular reactive oxygen species (ROS) were measured using $H_2DCFDA$. The proliferation and viability of VSMCs were inhibited by microtubule regulation. Additionally, microtubule-regulated and PDGF-BB-stimulated VSMCs increased the cleavage of caspase-3 more than only the microtubule-regulated condition, similar to that of LC3-II, implying autophagy. Inhibitory autophagy of microtubule-regulated and PDGF-BB-stimulated VSMCs resulted in low viability. However, enhancement of autophagy maintained survival through the reduction of ROS. These results suggest that the apoptosis of conditioned VSMCs is decreased by the blocking generation of ROS via the promotion of autophagy, and proliferation is also inhibited. Thus, promoting autophagy as a therapeutic target for vascular restenosis and atherosclerosis may be a good strategy.

• Polymer(Korea)
• /
• v.30 no.4
• /
• pp.357-361
• /
• 2006

Alpha-lipoic acid (ALA) synthesized in the body has virtues such as anti-oxidation, blood sugar regulation, appetite suppression, and anti-obesity, etc. ALA, which is also used as a drug, has a five-membered ring including disulfide and so easily losses bioavailability due to ring opening and subsequent polymerization by heat or ultraviolet. This report studies various conditions for ring opening polymerization. The ring opening starts above the melting point of ALA, but there was no temperature dependence above it. At $70^{\circ}C$, the degree of ring opening was proportional to reaction time and inversely proportional concentration. The degree of ring opening in acetic acid with UV for 1 hour reached the maximum conversion (70%). Most cleaved ALA changed into disulfide polymers, and the molecular weight of the polymers increased as the amount of ring opening increased.