• Journal of Microbiology and Biotechnology
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• 제25권7호
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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.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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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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• 제41권2호
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• pp.135-140
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• 2008

흡연은 활성 산소/질소종의 생성을 증가시켜 체내 산화적 스트레스를 증가시키고 폐의 염증을 유발한다. 이는 흡연자들의 체내 항산화 영양소들의 감소와 밀접한 관계를 가지는 것으로 알려져 있다. 본 연구에서는 항산화, 항염증 기능을 가진 비타민 E 중 식이에 많이 포함되어 있는 GT를 이용하여 흡연에 의한 항산화, 항염증 작용을 알아보기 위한 선행 연구로 고농도의 GT 식이가 혈액과 간, 폐의 AT, GT농도와 이들의 대샤산물인 CEHC 농도에 미치는 영향에 대하여 알아보고자 실시하였다. 연구 결과 고농도 GT 식이는 체내 혈액과 간, 폐 조직에 GT를 축적시키고 G-CEHC의 배설을 증가시켰다. 흡연은 대조군의 혈액과 폐 AT 농도는 증가시켰지만, 고농도 GT군의 혈액과 폐의 GT농도와 간의 G-CEHC의 농도를 감소 시켰다. 이러한 변화는 흡연에 의한 산화적 스트레스 상태에서 각기 다른 기능을 가진 조직의 요구량에 따라, 폐의 이용률을 높이기 위해 혈액으로 운반되는 양을 증가시키고 폐로의 운반을 임시적으로 증가시켜 조직의 AT, GT 농도를 선택적으로 조절하고 GT의 생체 이용률의 증가 때문이라 사료된다. 하지만 정확한 기전에 대한 연구들이 부족한 실정이므로 고농도의 GT 식이가 흡연에 의해 유도된 체내 산화적 스트레스와 폐의 염증 반응에 긍정적인 효과를 알아보기 위한 후속 연구가 절설히 필요한 것으로 사료된다.

• 핵의학기술
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• 제16권1호
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• pp.34-37
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• 2012

$^{18}F$-FDG 검사 시 한 bed 당 적정 스캔 시간을 정량 평가하기 위하여 Philips사 TF 64 PET/CT를 사용하였다. Phantom 실험과 환자를 대상으로 다양한 스캔 시간을 적용하며 평가를 하여 최적의 스캔 시간을 알고자 하였다. 20초 간격으로 총 5회의 서로 다른 bed 당 스캔 시간을 NEMA IEC body phantom과 환자들에게 적용하여 $^{18}F$-FDG 영상을 획득하였다. Phantom 실험의 경우 관심영역과 background의 방사능비를 8:1로 했다. 관심영역과 background 영역의 표준섭취계수(SUV)와 대조회복계수(CRC)가 계산되었다. 환자를 대상으로 한 연구에서는 평균$444{\pm}74$ MBq의 $^{18}F$-FDG가 환자에게 주사되었고 20명의 환자, 38개의 병소부위에서 표준섭취계수가 측정되었다. Background noise는 background 부분의 표준편차를 평균으로 나누어 계산되었다. Phantom 실험에서 두 그룹 (LT-60 [<60 sec]과 GT-60 [${\geq}60$ sec])을 비교한 결과 LT-60 그룹의 표준섭취계수(SUV)와 대조회복계수(CRC)가 (H1: 14.2 and 7.3, H2: 11.4 and 7.8, H3: 4.9 and 3.2), GT-60 그룹(H1: 8.9 and 2.8, H2: 8.2 and 5.0, H3: 2.0 and 1.6)에서 보다 높은 변이율을 보였다. 이는 한 bed 당 스캔시간이 60 sec 이상일 때 안정적인 표준섭취계수와 대조회복계수를 나타내는 것을 의미한다. 환자를 대상으로 한 연구에서 역시 표준섭취계수(SUV)와 대조회복계수(CRC)가 LT-60 그룹에서 높은 변이율을 보였다. PET/CT 영상에서 background noise는 bed 당 스캔시간이 증가함에 따라 감소하는 것으로 나타났다. 그리고 60 sec 이하 그룹에서는 높은 background noise로 인하여 높은 SUV와 CRC 변이율을 보였다. 따라서 PET/CT 영상의 질을 고려할 때 1 bed 당 스캔시간은 최소 60 sec 이상이어야 한다.

• Journal of Periodontal and Implant Science
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• 제53권5호
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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.

• 한국환경보건학회지
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• 제30권3호
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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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• 제3권5호
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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.

• 한국자원식물학회지
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• 제17권3호
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• pp.352-357
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• 2004

쓴메밀의 종자의 발아율은 발아온도의 증가와 더불어 증가되는 경향을 보이며, 초장은 3$0^{\circ}C$에서 가장 좋은 생육을 보이고 그 다음으로 $25^{\circ}C$ ＆gt; 2$0^{\circ}C$ ＆gt; 15$^{\circ}C$순으로 온도가 높을수록 초장의 생육이 왕성하였다. 온도처리에 따른 쓴메밀의 초장은 $25^{\circ}C$ ＆gt; 2$0^{\circ}C$ ＆gt; 15$^{\circ}C$ ＆gt; 3$0^{\circ}C$ 순으로 좋았으며, 엽수도 2$0^{\circ}C$ 및 $25^{\circ}C$에서 15$^{\circ}C$ 및 3$0^{\circ}C$보다 다소 증가하는 경향을 보였다. 일장처리에서 16시간 처리구를 제외하면 일장이 길어질수록 초장, 엽수 및 건물 중은 증가하는 경향을 보였으나, 루틴함량과 일장에 대한 일정한 경향은 없었다. 쓴메밀은 초장, 엽수, 분지수 및 경직경 등의 지상부 생육이 단메밀의 생육에 비해 왕성하였다. 특히 엽수에서 단메밀이 62.3에 비해 174.1로 약 2.8배 높았으며, 개체당 종실의 수량은 약 5.4배, 종실무게는 2.1배로 큰 폭의 차이 나타냈다. 쓴메밀 종실의 루틴함량이 1469.8로 단메밀의 22.6보다 약 60배 이상 높게 함유되어 있다.

• Korean Journal of Acupuncture
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• 제21권1호
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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.

• 대한임상검사과학회지
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• 제45권4호
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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.