• Journal of Life Science
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• v.21 no.1
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• pp.119-126
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• 2011

There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

• Journal of Life Science
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• v.22 no.10
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• pp.1359-1370
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• 2012

Arabidopsis AtERF71/HRE2, a transcription activator, is located in the nucleus and is involved in the signal transduction of low oxygen and osmotic stresses. In this study, microarray analysis using AtERF71/HRE2-overexpressing transgenic plants was performed to identify genes downstream of AtERF71/HRE2. A total of 161 different genes as well as AtERF71/HRE2 showed more than a twofold higher expression in AtERF71/HRE2-overexpressing transgenic plants compared with wild-type plants. Among the 161 genes, 24 genes were transcriptional regulators, such as transcription factors and DNA-binding proteins, based on gene ontology annotations, suggesting that AtERF71/HRE2 is an upstream transcription factor that regulates the activities of various downstream genes via these transcription regulators. RT-PCR analysis of 15 genes selected out of the 161 genes showed higher expression in AtERF71/HRE2-overexpressing transgenic plants, validating the microarray data. On the basis of Genevestigator database analysis, 51 genes among the 161 genes were highly expressed under low oxygen and/or osmotic stresses. RT-PCR analysis showed that the expression levels of three genes among the selected 15 genes increased under low oxygen stress and another three genes increased under high salt stress, suggesting that these genes might be downstream genes of AtERF71/HRE2 in low oxygen or high salt stress signal transduction. Microarray analysis results indicated that AtERF71/HRE2 might also be involved in the responses to other abiotic stresses and also in the regulation of plant developmental processes.

• Journal of Life Science
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• v.20 no.5
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• pp.729-735
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• 2010

In our previous study, it was reported that an herbal mixture, SH21B, inhibits fat accumulation and adipogenesis both in vitro and in vivo models of obesity. SH21B is a mixture composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd, and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). The aim of this study was to investigate the detailed molecular mechanisms of the effects of SH21B on various regulators of the adipogenesis pathway. During the adipogenesis of 3T3-L1 cells, SH21B significantly decreased the expression levels of central transcription factors of adipogenesis, such as peroxisome proliferator-activated receptor (PPAR)$\gamma$ and CCAAT/enhancer binding protein (C/EBP)$\alpha$. To elucidate the detailed molecular mechanism of the anti-adipogenic effects of SH21B, we examined the expression levels of the various pro-adipogenic or anti-adipogenic regulators of adipogenesis upstream of $PPAR{\gamma}$ and C/$EBP{\alpha}$. The mRNA levels of Krox20 and Kruppel-like factor (KLF) 15, which are pro-adipogenic regulators, were significantly down-regulated by SH21B treatment, whereas the mRNA levels of C/$EBP{\gamma}$ and KLF5 were not changed. KLF2 and C/EBP homologous protein (CHOP), which are anti-adipogenic regulators, were significantly up-regulated by SH21B treatment. These results suggest that the molecular mechanism of the anti-adipogenic effect of SH21B involves both the down-regulations of pro-adipogenic regulators, such as Krox20 and KLF15, and the up-regulations of anti-adipogenic regulators, such as KLF2 and CHOP, which results in the suppression of central transcription factors of adipogenesis including $PPAR{\gamma}$ and C/$EBP{\alpha}$.

• Restorative Dentistry and Endodontics
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• v.36 no.4
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• pp.300-305
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• 2011

Objectives: The aim of this study was to evaluate the various NiTi rotary instruments regarding their ability to provide a circular apical preparation. Materials and Methods: 50 single canal roots were selected, cut at the cementodentinal junction and the coronal 1/3 of the canals was flared using Gates Glidden burs. Samples were randomly divided into 5 experimental groups of 10 each. In group I, GT files, Profile 04 and Quantec #9 and #10 files were used. In Group II Lightspeed was used instead of Quantec. In Group III, Orifice shaper, Profile .06 series and Lightspeed were used. In Group IV, Quantec #9 and #10 files were used instead of Lightspeed. In Group V, the GT file and the Profile .04 series were used to prepare the entire canal length. All tooth samples were cut at 1 mm, 3 mm and 5 mm from the apex and were examined under the microscope. Results: Groups II and III (Lightspeed) showed a more circular preparation in the apical 1mm samples than the groups that used Quantec (Group I & IV) or GT files and Profile .04 series.(Group V)(p < 0.05) There was no significant difference statistically among the apical 3, 5 mm samples. In 5 mm samples, most of the samples showed complete circularity and none of them showed irregular shape. Conclusions: Lightspeed showed circular preparation at apical 1 mm more frequently than other instruments used in this study. However only 35% of samples showed circularity even in the Lightspeed Group which were enlarged 3 ISO size from the initial apical binding file (IAF) size. So it must be considered that enlarging 3 ISO size isn't enough to make round preparation.

• Journal of Sensor Science and Technology
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• v.10 no.1
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• pp.62-70
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• 2001

The ZnSe sample grown by chemical bath deposition (CBD) method were annealed in Ar gas at $45^{\circ}C$. Using extrapolation method of X-ray diffraction pattern, it was found to have zinc blend structure whose lattice parameter $a_o$ was $5.6687\;{\AA}$. From Hall effect, the mobility was likely to be decreased by impurity scattering at temperature range from 10 K to 150 K and by lattice scattering at temperature range from 150 K to 293 K. The band gap given by the transmission edge changed from $2.700{\underline{5}}\;eV$ at 293 K to $2.873{\underline{9}}\;eV$ at 10 K. Comparing photocurrent peak position with transmission edge, we could find that photocurrent peaks due to excition electrons from valence band, ${\Gamma}_8$ and ${\Gamma}_7$ and to conduction band ${\Gamma}_6$ were observed at photocurrent spectrum. From the photocurrent spectra by illumination of polarized light on the ZnSe thin film, we have found that values of spin orbit coupling splitting ${\Delta}so$ is $0.098{\underline{1}}\;eV$. From the PL spectra at 10K, the peaks corresponding to free bound excitons and D-A pair and a broad emission band due to SA is identified. The binding energy of the free excitons are determined to be $0.061{\underline{2}}\;eV$ and the dissipation energy of the donor -bound exciton and acceptor-bound exciton to be $0.017{\underline{2}}\;eV$, $0.031{\underline{0}}\;eV$, respectively.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.699-720
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• 1995

This study was conducted to evaluate the effect of benzalkonium chloride solution as a wetting agent instead of water on dentin bonding with NTG-GMA/BPDM system (All-bond 2, Bisco.) and DSDM system (Aelitebond, Bisco.). Benzalkonium chloride solution is a chemical disinfectant widely used in medical and dental clinics for preoperative preparation of skin and mucosa due to its strong effect of cationic surface active detergent. Eighty freshly extracted bovine lower incisor were grinded labially to expose flat dentin surface, and then were acid-etched with 10 % phosphoric acid for 15 second, water-rinsed, and dried for 10 second with air syringe. The specimens were randomly divided into 8 groups of 10 teeth. The specimens of control group were remoistured with water and the specimens of experimental groups were remoistured with 0.1 %, 0.5 %, and 1.0 % benzalkonium chloride solution respectively. And then, the Aelitefil composite resin was bonded to the pretreated surface of the specimens by use of All-bond 2 dentin bonding system or Aelitebond dentin bonding system in equal number of the specimens. The bonded specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured, the mode of failure was observed, the fractured dentin surface were examined under scanning electron microscopy, and FT-IR spectroscopy was taken for the purpose of investigating the changes of the dentin surface pretreated with benzal konium chloride solution followed by each primer of the dentin bonding systems. The results were as follows : In the group of bonding with NTG-GMA/BPDM dentin bonding agent(All-bond 2), higher tensile bond strength was only seen in the experimental group remoistured with 0.1 % benzal konium chloride solution than that in water-remoistured control group(p<0.05). In the group of bonding with DSDM dentin bonding agent (Aelitebond), no significant differences were seen between the control and each one of the experimental group(p<0.05). Higher tensile bond strength were seen in NTG-GMAIBPDM dentin bonding agent group than in DSDM dentin bonding agent group regardless of remoistur ization with benzal konium chloride solution. On the examination of failure mode, cohesive and mixed failure were predominantly seen in the group of bonding with NTG-GMAIBPDM dentin bonding agent, while adhesive failure was predominantly seen in the group of bonding with DSDM dentin bonding agent. On SEM examination of fractured surfaces, no differences of findings of primed dentin surface between the groups with and without remoisturization with benzal konium chloride solution. FT-IR spectroscopy taken from the control and the experimental group reve::.led that some higher absorbance derived from the primers binding to dentin surface was seen at the group pretreated with 0.1 % benzal konium chloride solution than at the control group of remoisturizing with water.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.243-247
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• 2006

The biological activity of functional food source with thyme extracts were examined. Total phenol contents in the 60% ethanol extracts $(26.8{\pm}0.35\;mg/g)$ with thyme leaf was higher than water extracts $(25.7{\pm}0.20\;mg/g)$. This HPLC analysis is significant in that physiological activity is related with phenolic compound content such as rosemarinic acid, quercetin and chlorogenic acid. Electron donating ability was shown as 90.1% in the water extracts and 77.7% in the 60% ethanol extracts. Antioxidant protection factor of 60% ethanol extracts was higher than water extracts. Helicobacter pylori of the water extracts from thyme leaves did not have antimicrobial activity, but the 60% ethanol extracts revealed the high antimicrobial activity as 9 mm of clear zone in $50\;{\mu}g/ml$ of phenol content, 10 mm in $100\;{\mu}g/ml$, 13 mm in $150\;{\mu}g/ml$ and 16 mm in $200\;{\mu}g/ml$, respectively. Angiotensin converting enzyme inhibition activity showed no inhibition activity in 60% ethanol extracts but 39.9% inhibition activity in water extracts. Xanthine oxidase inhibition activity showed high inhibition activity at 73.5% in water extracts and 100% in 60% ethanol extracts. The result suggests the development of phenol compound in thyme as anti Helicobacter pylori, antioxidant and anti-gout agents.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.153-161
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• 2002

Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

• Parasites, Hosts and Diseases
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• v.33 no.3
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• pp.201-210
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• 1995

This study aims to assess the possible strain-dependent variations in detection of ToxopLosmn antigens and antibodies. The virulent RH strain or avirulent Beverley strain of T gondii were injected into mice, intraperitoneally, and their antigens, antibodies and parasites were identified from the blood or tissues: liver, brain and spleen by ELISA, Western blot and PCR. In mice infected with RH strain, circulating antigens and parasitemia were first detected from 2 days after infection, and ToxopIasma DNA were found in the blood, liver, brain and spleen from 3 days after infection. It was impossible to detect specific IgM and IgG antibodies to T gondij and any specific band was not found by Western blot. In mice infected with Beverley strain, circulating antigens were detected between day 10 and day 35. The Toxoplusma DNA was found in the blood and liver from day 15 until day 60, and in the brain from day 20. But Toxoplosma DNA in the spleen were mainly detected between day 10 and day 30. The IgM antibodies were first appeared on day 10 post-infection, and were noted obviously increased between day 15 and 25. The IgG antibodies were first detected on day 15, and showed progressively increased titers. The antibody binding bands were specific according to infection period. Sera from mice infected with Beverley strain reacted mainly with the antigen of 27.5-kDa and 32.5-kDa. In conclusion, mice infected with RH strain revealed Toxoplosma antigens strongly, but not antibodies. However. mice infected with Beverley strain revealed both the Toxoplasma antigens and antibodies. The present results showed that immune responses are different between avirulent and virulent T gonnii.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.994-1004
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• 2005

The dihydrofolate reductase (dhfr) promoter contains cis-acting element for the transcription factors Spl and E2F. Transcription of dhfr gene shows maximal activity during the Gl/S phase of cell cycle. The member of the Spl transcriptional factor family can act as both negative and positive regulators of gene expression. There was a report that Spl-Rb and E2F4-pl30 complexes cooperate to establish stable repression of dhfr gene expression in CHOC400 cells. Here, we examined the role of HDAC in dhfr, cyclin E, and cyclin A gene regulation using the histone deacetylation inhibitor, trichostatin A (TSA) in U2OS and C33A cells, a Rb-positive human osteosarcoma cell line, and a Rb-negative cervical carcinoma cell line, respectively. When the dhfr promoter constructs were applied in U2OS cells, TSA markedly stimulated over 14-fold of dhfr promoter activity through dhfr-Spl sites by the deletion of an E2F element. In contrast, the deletion of dhfr-Spl binding sites completely abolished promoter stimulation by TSA. The dhfr promoter activity including dhfr-Spl sites increased only 2-fold in C33A cells. Promoter activity containing only dhfr-E2F site did not have much effect by the treatment of TSA in both U2OS and C33A cells. On the other hand, treatment with TSA induced significantly mRNA expression of dhfr and cyclin E, whereas levels of cyclin A decreased in U2OS cells, but had no effect in C33A cells. These results indicate that TSA have contradictory effect, activation of dhfr and cyclin E genes on Gl phase, and down-regulation of cyclin A on G2 phase through transcriptional regulation in U2OS cells.