• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1070-1083
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• 2015

A gene (GT-SM3B) encoding a thermostable secreted oligoendopeptidase (GT-SM3B) was cloned from the thermophile Geobacillus thermoleovorans DSM 15325. GT-SM3B is 1,857 bp in length and encodes a single-domain protein of 618 amino acids with a 23-residue signal peptide having a calculated mass of 67.7 kDa after signal cleavage. The deduced amino acid sequence of GT-SM3B contains a conservative zinc metallopeptidase motif (His⁴⁰⁰-Glu⁴⁰¹-X-XHis⁴⁰⁴). The described oligopeptidase belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepF_Ba from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases. Secretory production of GT-SM3B was used, exploiting successful oligopeptidase signal peptide recognition by Escherichia coli BL21 (DE3). The recombinant enzyme was purified from the culture fluid. Homodimerization of GT-SM3B was determined by SDS-PAGE. Both the homodimer and monomer were catalytically active within a pH range of 5.0–8.0, at pH 7.3 and 40℃, showing the K_m, V_max, and k_cat values for carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala-OH peptidolysis to be 2.17 ± 0.04 × 10^-6 M, 2.65 ± 0.03 × 10^-3 µM/min, and 5.99 ± 0.07 s^-1, respectively. Peptidase remained stable at a broad pH range of 5.0–8.0. GT-SM3B was thermoactive, demonstrating 84% and 64% of maximum activity at 50℃ and 60℃, respectively. The recombinant oligopeptidase is one of the most thermostable M3B peptidase, retaining 71% residual activity after incubation at 60℃ for 1 h. GT-SM3B was shown to hydrolyze a collagenous peptide mixture derived from various types of collagen, but less preferentially than synthetic hexapeptide. This study is the first report on an extracellular thermostable metallo-oligopeptidase.

• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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• pp.411.1-411.1
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• 2014

This study evaluates the utility of an antibacterial microneedle composed of green tea extract (GT) and hyaluronic acid (HA), for the efficient delivery of GT. These microneedles have the potential to be a patient-friendly method for the conventional sustained release of drugs. In this study, a fabrication method using a mold-based technique to produce GT/HA microneedles with a maximum area of ${\sim}60mm^2$ with antibacterial properties was used to manufacture transdermal drug delivery systems. Fourier transform infrared (FTIR) spectrometry was carried out to observe the potential modifications in the microneedles, when incorporated with GT. The degradation rate of GT in GT/HA microneedles was controlled simply by adjusting the HA composition. The effects of different ratios of GT in the HA microneedles were determined by measuring the release properties. In HA microneedles loaded with 70% GT (GT70), a continuous higher release rate were sustained for 72 h. The in vitro cytotoxicity assays demonstrated that GT/HA microneedles are not generally cytotoxic to chinese hamster ovary cells (CHO-K1), human embryonic kidney cells (293T), and mouse muscle cells (C2C12), which were treated for 12 and 24 h. Antimicrobial activity of the GT/HA microneedles was demonstrated by ~95% growth reduction of gram negative [Escherichia coli (E. coli), Pseudomonas putida (P. putida) and Salmonella typhimurium (S. typhimurium)] and gram positive bacteria [Staphylococcus aureus (S. Aureus) and Bacillus subtilis (B. subtilis)], with GT70. Furthermore, GT/HA microneedles reduced bacterial growth in the infected skin wound sites and improved skin wound healing process in rat model.

• Journal of Nutrition and Health
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• v.41 no.2
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• pp.135-140
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• 2008

Cigarette smoke (CS) induces oxidative and nitrosative stress to the respiratory tract (RT) via both oxidants contained in CS and by CS-induced activation of RT inflammatory-immune pro-oxidant processes. CS exposure has been associated with reduced levels of plasma micronutrient antioxidants, in part due to an increased utilization and turnover of alpha-tocopherol (AT). It has been suggested that gamma-tocopherol (GT) may have an expanded spectrum of antioxidant activation compared to alpha-tocopherol (AT). In order to investigate effects of high GT supplementation as compared to AT, C57 BL/6 mice were fed control AT (35 mg/kg diet) or high GT (1,000 mg/kg diet) diet for 8-10 weeks and then exposed to 60 $mg/m^3$ CS, 6 hr/day for 3 days. AT and GT levels and their metabolites were measured at endpoints. High GT supplementation significantly reduced AT levels in plasma, liver and lung compared to AT. CS increased levels of AT and GT in plasma and lung of control AT group but decreased GT levels in lung of high GT supplemented group. Moreover, CS significantly decreased GT metabolite, gamma-CEHC. The results suggest that high GT supplementation have selective modulation of concentrations of vitamin E and its metabolite in plasma and lung but not in liver against in vivo CS exposure.

• The Korean Journal of Nuclear Medicine Technology
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• v.16 no.1
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• pp.34-37
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• 2012

Purpose: To verify the optimal scan time per bed for clinical application, we evaluated the quality of $^{18}F$-FDG images with varying scan times in a phantom and 20 patients with 38 lesions using a Philips (TOF) PET/CT scanner. Materials and Methods: The PET/CT images of a NEMA IEC body phantom and 20 patients (16 males, 4 females) were acquired for 5 different scan times of 20-100 sec per bed with intervals of 20 sec. The activity ratio of hot spheres (diameter of 17 [H1], 22 [H2] and 28 [H3] mm) to the background region in the IEC body phantom was 8-to-1. The contrast recovery coefficient (CRC) and standard uptake value (SUV) based on ROIs of hot spheres and background region were calculated. The noise in each background region was estimated as the ratio of SD of counts to the mean counts in the background region. On the patient image, the injected dose of $^{18}F$-FDG was $444{\pm}74$ MBq and the SUVs in the 38 hot lesions were measured. Results: The two scan time groups (LT-60 [<60 sec] and GT-60 [${\geq}60$ sec]) were compared. In the phantom study, the coefficient of deviations (CVs, %) of CRC and SUV in LT-60 (H1: 14.2 and 7.3, H2: 11.4 and 7.8, H3: 4.9 and 3.2) were higher than GT-60 (H1: 8.9 and 2.8, H1: 8.2 and 5.0, H3: 2.0 and 1.6). In the patient study, the mean CV of CRC and SUV in LT-60 (4.0) was higher than GT-60 (1.2). Conclusion: This study showed that noise increased as the scan time decreased. High noise for the scan time <60 sec per bed yielded high variation of SUV and CRC. Therefore, considering PET/CT image quality, the scan time per bed in the TOF PET/CT scanner should be at least ${\geq}60$ sec.

• Journal of Periodontal and Implant Science
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• v.53 no.5
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• pp.362-375
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• 2023

Purpose: The aim of this study was to investigate the relationships between gingival thickness (GT) and keratinized gingiva width (KGW), papilla height (PH), and crown ratio (CR) by employing transgingival probing and an intraoral scanner (IOS). Methods: This cross-sectional study examined 360 maxillary anterior teeth from 60 patients. GT was assessed using transgingival probing with an endodontic spreader. KGW, CR, and PH were measured using an IOS. One-way analysis of variance, the Student's t-test, and Spearman correlation coefficients were employed for statistical analysis. Results: Higher GT was significantly associated with thinner KGW in the central region (P=0.019). There was no statistically significant difference in GT between teeth (P=0.06). PH was lower in lateral teeth than in canines (P=0.047), with a PH of 2.99 mm in lateral teeth. The KGW was narrower in canines than in central teeth (P=0.007). A moderate correlation was observed between KGW and PH in the central region (P=0.01), while a weak negative correlation was found between KGW and CR (P=0.043). Conclusions: A moderate negative correlation was found between GT and KGW, as well as between PH and KGW in central teeth. In contrast, a weak negative correlation existed between CR and KGW. The PH (2.99 mm) was lower in lateral teeth than in canines. The traditional paradigm, which suggests a positive correlation between KGW and GT, was reevaluated by measuring KGW using an IOS.

• Journal of Environmental Health Sciences
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• v.30 no.3
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• pp.230-238
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• 2004

The adverse health effects of a number of environment pollutions are related to the formation of free radicals. Induction of antioxidant defensive system in the response to an oxidative attack is an essential element of the cell to survive. CYP2E1 is easily induced by organic solvents and induces continuous formation of reactive oxygen species (ROS). ${\gamma}$-Glutamyltranspeptidase (${\gamma}$GT) plays an important role in glutathione metabolism and xenobiotic detoxification. To evaluate the characteristic of oxidative stress which induces GGT expression and to understand human antioxidant defensive response against oxidative stress induced by CYP2E1, we studied regulation of ${\gamma}$GT enzyme expression in response to various oxidative stresses in human HepG2 cells. The ${\gamma}$GT activity was not modified after exposure of acute oxidative stress inducing agents (ferric nitrilotriacetate, cumene hydroperoxide, ADP-Fe, O-tetradecanoylphorbol-13-acetate, tumor necrosis factor-alpha). To induce continuous exposure of cells to ROS, HepG2 cells were transfected by human CYP2E1 gene transiently. The CYP2E1 activity was verified with chlorzoxazone hydroxylation. Transfection of CYP2E1 showed continuous 60% increase in intracellular ROS and 240 % increase in microsomal ROS. CYP2E1 overexpressing cells showed increased ${\gamma}$GT activity (2.5-fold). The observed enhancement of ${\gamma}$GT activity correlated with a significant increase of ${\gamma}$GT mRNA (2.1-fold). Treatment with antioxidant strongly prevented the increase in ${\gamma}$GT activity. The CYP2E1 overexpression did not modify toxicity index and increased glutathione levels. These results show that continuous exposure of cells to ROS produced by CYP2E1 up-regulates ${\gamma}$GT; This may be one of the adaptive antioxidant responses of cells to oxidative insult. Present study also suggests that the induction of ${\gamma}$GT could be used as a marker of oxidative stress induced by exposure to organic solvents.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.5
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• pp.501-505
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• 1999

Tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin biosynthesis, is primarily expressed in serotonergic neurons of the raphe nuclei. Simple tandem repeat polymorphisms, typically one to four nucleotides long, are tandemly repeated several times and often characterized by many alleles. To identify the presence of polymorphic repeats, we sequenced the 5'-upstream region of the mouse TPH gene. For the detection of any allelic variants, polymerase chain reaction, nonisotopic single-strand conformation polymophism, and DNA sequencing analyses of the tandem repeat sequences were performed using genomic DNA extracted from 60 ICR mice. Two dinucleotide repeats, $5'-(AC/TG)_{22}-3'$ and $5'-(GT/CA)_{17}3',$ were identified at approximately - 5.7 kb and - 3.4 kb upstream from the transcriptional initiation site of the mouse TPH gene, respectively. Minor allelic variants, $5'-(AC/TG)_{21}-3'$ and $5'-(GT/CA)_{18}-3',$ were observed in heterozygous pairs from 3 of 60 and 1 of 60 ICR mice, respectively. The identification of these microsatellites in the mouse TPH promoter raises the possibility that identical and/or other polymorphic sequences might exist in the upstream region of the human TPH gene.

• Korean Journal of Plant Resources
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• v.17 no.3
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• pp.352-357
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• 2004

This study was carried out to investigate the effects of temperature and photoperiod on the growth of Fagopyrum tataricum. It showed a tendency to promote the germination rate and plant height of tartary buckwheat under high temperature. Plant height and number of leaves were promoted in the order of 25 ＆gt; 20 ＆gt; 15 ＆gt; 3$0^{\circ}C$ under the different temperature after 30 days of the planting. In the 10, 12, 14hr photoperiods at 2$0^{\circ}C$, plant height, number of leaves and dry weight were increased as much as photoperiod became long except by 16hr. Rutin contents was not regular both common and tartary buckwheat under the photoperiods. The rutin content in leaves was higher than stem. Compared to yield of tartary and common buckwheat, plant height, number of leaves, number of branch and stem diameter of tartary buckwheat increased more than common buckwheat. Especially, number of leaves was increased about 2.5 times more than common buckwheat. And there was a difference in about 2.1 times in grain weight and about 5.4 times in number of grains. Rutin content was 1469.8mg/l00g in grain of tartary buckwheat. It was about 60 times higher than 22mg/100g in grain of common buckwheat.

• Korean Journal of Acupuncture
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• v.21 no.1
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• pp.51-60
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• 2004

Objectives : This research was performed to investigate the effect of herbal acupuncture(Ephedra sinica and Green tea) at Umnungchon(SP9) and Pungnyung(ST40) on weight gain, food intakes, food efficiency ratios, HDL-cholesterol, triglyceride, free fatty acid on rats fed high fat diet for 5weeks. Methods : Experimental groups were divided into normal group(Sham), high fat diet group(Con), high fat diet and Ephedra sinica-herbal acupuncture in SP9 ST40 treated group(ES), high fat diet and Green tea-herbal acupuncture in SP9 ST40 treated group (GT), high fat diet and Ephedra sinica+Green tea-herbal acupuncture in SP9 ST40 treated group(ES+GT). Herbal acupuncture was bilaterally treated at the level of 19.125mg/ml per 2 days during 5 weeks. Results : Body weight, feed intake and feed efficiency ratio were decreased in Sham, ES, ES+GT, compared with control group. The level of serum HDL-cholesterol was increased in sham, tender to increase in ES, ES+GT. The change on that of serum triglyceride was decreased in sham, ES, ES+GT and free fatty acid was decrease in sham, ES+GT. Conclusions : Ephedra sinica with Green tea-herbal acupuncture in SP9 ST40 can control the Body weight, feed intake and feed efficiency ratio, HDL-cholesterol, triglyceride, free fatty acid.

• Korean Journal of Clinical Laboratory Science
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• v.45 no.4
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• pp.149-153
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• 2013

Candidiasis is a fungal infection of the most common causes; generally, opportunistic infections occur often in patients with weakened immune systems. Because of high rates in fungal infection patients and increasing frequency of being isolated from clinical materials, quickly identifying of Candida albicans is critical. By identifying 404 yeast cell strains of referred samples via API 20C kits, NGL and PRO tests and Germ tube (GT) test were conducted and compared. In the 3.0 McFarland yeast cells, 0.1% ${\rho}-nitrophenyl-N-acetyl-{\beta}-D-galactosaminide$ (NGL) and 0.04% ${\small{L}}$-proline ${\beta}$-naphtylamide (PRO) were each put in test tubes and incubated at $35^{\circ}C$ for 15, 30, 60 and 90 minutes. Afterwards, 1 drop of 2% NaOH was applied, and if the color turned yellow; it was positive for NGL test. Afterwards, 1% ${\rho}$-dimethylaminocinnamaldehyde was applied, and if the upper layer turned pink or red, it was positive for PRO test. NGL and PRO tests were conducted for all C. albicans and identified accurately within 30 minutes. In NGL, PRO test, false-positive, negative were not seen, whereas, GT test showed false-positive in 1 strain and false-negative in 3 strains. Therefore, sensitivity and specificity of NGL, PRO tests were 100% and 99.5%, respectively, and positive and negative predictive rate were 99.5% and 100%, respectively. However, GT test sensitivity and specificity were 98.5% and 99.5%, respectively, and positive and negative predictive rates were 99.5% and 98.5%, respectively. In conclusion, NGL, PRO tests are better than GT tests for sensitivity and specificity, therefore, these reliable tests will be useful in clinical laboratories.